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|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
PMC11277193_p75
|
PMC11277193
|
sec[2]/sec[5]/p[1]
|
3.6. Dose Escalation and Reinduction in UC
| 3.890625 |
biomedical
|
Study
|
[
0.94140625,
0.05706787109375,
0.001689910888671875
] |
[
0.82080078125,
0.173583984375,
0.0021953582763671875,
0.0031833648681640625
] |
The long-term extension UNIFI trial evaluated the efficacy of dose adjustment from q12w to q8w in 523 UC patients at week 56. Clinical remission at week 56 was observed in 70.0% of patients shortening the administration interval. Because most patients were already in remission at optimization, or had mild activity, the real value of these data is debatable .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277193_p76
|
PMC11277193
|
sec[2]/sec[5]/p[2]
|
3.6. Dose Escalation and Reinduction in UC
| 4.105469 |
biomedical
|
Study
|
[
0.93701171875,
0.0618896484375,
0.0008597373962402344
] |
[
0.96337890625,
0.03179931640625,
0.0014095306396484375,
0.003467559814453125
] |
Intensification, every 4 or 6 weeks, was required after a median time of 95 days in 42.6% of 108 UC patients . Twelve to sixteen weeks after intensification, 55.0% achieved remission and 67.5% achieved response (reduction in SCCAI/Mayo by 3 points from baseline). After a median follow-up of 230 days after dose escalation, 56.3% had endoscopic improvement and FC reduction ( Table 2 , study design reported in Supplementary Table S1 ) .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277193_p77
|
PMC11277193
|
sec[2]/sec[5]/p[3]
|
3.6. Dose Escalation and Reinduction in UC
| 4.074219 |
biomedical
|
Study
|
[
0.90380859375,
0.09515380859375,
0.0009450912475585938
] |
[
0.974609375,
0.0188140869140625,
0.0015001296997070312,
0.005115509033203125
] |
This was confirmed in a small retrospective cohort of 23 patients. During the 12 months follow-up, dose adjustment was required in 13 patients (56%). Three (13%) underwent intravenous UST reinduction and ten (43%) dose escalation (eight by shortening the interval between doses and two by switching to intravenous USK administration). After 52 weeks from dose adjustment, clinical remission was reached in 79% of the patients, and normalization of CRP and FC in 80% and 40%, respectively. Five of the six patients who underwent endoscopic examination following adjustment showed a Mayo subscore ≤ 1 .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277193_p78
|
PMC11277193
|
sec[2]/sec[5]/p[4]
|
3.6. Dose Escalation and Reinduction in UC
| 4.164063 |
biomedical
|
Study
|
[
0.9931640625,
0.006710052490234375,
0.00025534629821777344
] |
[
0.99658203125,
0.00206756591796875,
0.000804901123046875,
0.0005598068237304688
] |
A real-world retrospective cohort study in 40 UC patients undergoing dose intensification q4w or q6w provides similar data. More than 50% of patients achieved corticosteroid-free clinical remission, defined as HBI < 5 and no use of systemic steroids within 30 days, at 1 year and more than 40% at month 24. Interestingly, more than 80% of the UC patients in corticosteroid-free remission at week 12 maintained remission at week 24. Endoscopic response was observed in 55% of those patients undergoing the procedure while on UST .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277193_p79
|
PMC11277193
|
sec[2]/sec[5]/p[5]
|
3.6. Dose Escalation and Reinduction in UC
| 4.109375 |
biomedical
|
Study
|
[
0.98779296875,
0.01192474365234375,
0.0004582405090332031
] |
[
0.994140625,
0.004596710205078125,
0.0007677078247070312,
0.0006604194641113281
] |
More data were derived from a small, mixed cohort of 42 IBD patients (8 UC) undergoing dose escalation q4w or q6w or dose intensification followed by IV reinduction. Fifty-two weeks later, 14 patients (33.3%) achieved corticosteroid-free clinical remission, biochemical remission and endoscopic healing. No significant advantage was provided by reinduction ( p = 0.99) .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277193_p80
|
PMC11277193
|
sec[2]/sec[5]/p[6]
|
3.6. Dose Escalation and Reinduction in UC
| 1.954102 |
clinical
|
Clinical case
|
[
0.426025390625,
0.56884765625,
0.004917144775390625
] |
[
0.17626953125,
0.335693359375,
0.0076141357421875,
0.48046875
] |
A few case reports also suggested the efficacy of dose escalation, with one case documenting clinical improvement following a q3w injection regimen. No adverse events were reported in the 6-month follow-up .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277193_p81
|
PMC11277193
|
sec[2]/sec[5]/p[7]
|
3.6. Dose Escalation and Reinduction in UC
| 3.777344 |
biomedical
|
Review
|
[
0.99853515625,
0.00103759765625,
0.000553131103515625
] |
[
0.15673828125,
0.03765869140625,
0.8046875,
0.00093841552734375
] |
Despite the lack of hard data on UST dose intensification and reinduction in UC, available evidence suggests that, similarly to CD, this may represent a viable strategy in primary nonresponse and secondary loss of response. Prospective studies and larger series are needed to confirm preliminary evidence and identify specific subpopulations that would benefit from the differing optimization strategies.
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277193_p82
|
PMC11277193
|
sec[3]/p[0]
|
4. Conclusions
| 3.802734 |
biomedical
|
Other
|
[
0.99658203125,
0.0027141571044921875,
0.0005846023559570312
] |
[
0.034027099609375,
0.9345703125,
0.028533935546875,
0.0028209686279296875
] |
Ustekinumab is a fully human antibody preventing the bond of the subunit p40 of IL-12 and IL-23 with the receptor proteins expressed on immune cells. It hinders IL-12 and IL-23 biological activity, affecting T-cell differentiation toward Th1 lineage and Th17 cell-related immune response . First licensed for psoriasis and psoriatic arthritis, UST received regulatory approval for CD in 2016 and UC in 2019 .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277193_p83
|
PMC11277193
|
sec[3]/p[1]
|
4. Conclusions
| 4.046875 |
biomedical
|
Review
|
[
0.99169921875,
0.005245208740234375,
0.00308990478515625
] |
[
0.00885009765625,
0.0015954971313476562,
0.9892578125,
0.00034308433532714844
] |
UST proved effective in inducing and maintaining clinical response and remission in IBD, and is presently considered a viable second-line treatment. Head-to-head comparisons of different biologics are scarce, and evaluations mostly derive from indirect comparison or retrospective studies. Available evidence suggests that anti-TNFs are superior to UST as first-line therapy in CD. The efficacy of UST as second-line therapy ranges between 35% and 50% . Figures are comparable to other second-line therapies in terms of the response, remission and adverse event rates in uncomplicated CD . Efficacy is comparable to other biologics also in fistulizing CD while experience is poor in postoperative recurrence.
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277193_p84
|
PMC11277193
|
sec[3]/p[2]
|
4. Conclusions
| 3.980469 |
biomedical
|
Review
|
[
0.9970703125,
0.0017528533935546875,
0.0010128021240234375
] |
[
0.06390380859375,
0.00222015380859375,
0.93359375,
0.00044155120849609375
] |
Less data are available in UC patients compared to CD, but no major differences have been reported versus other second-line biologic agents . Network metanalysis reported similar efficacy of UST and TOF in UC patients, both proving superior to VDZ . Again, conclusions were drawn on the base of statistical tools, and need confirmation in real-life, head-to-head studies. Despite low or moderately-low evidence, UST was recommended, as well as TNF agents VDZ and TOF, by the 2022 ECCO consensus in patients with moderate-to-severe UC and inadequate response or intolerance to conventional therapy .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277193_p85
|
PMC11277193
|
sec[3]/p[3]
|
4. Conclusions
| 3.566406 |
biomedical
|
Other
|
[
0.99658203125,
0.0027713775634765625,
0.0006847381591796875
] |
[
0.07659912109375,
0.5107421875,
0.41015625,
0.0025234222412109375
] |
Like all other biologics and small molecules, UST is burdened by a proportion of primary nonresponse and loss of response during maintenance therapy. Dose escalation and reinduction strategies before switching out of class have been reported in an increasing number of studies, but the best therapeutic strategy and its positioning in the treatment workup of IBD is still undefined.
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277193_p86
|
PMC11277193
|
sec[3]/p[4]
|
4. Conclusions
| 3.462891 |
biomedical
|
Study
|
[
0.99072265625,
0.008087158203125,
0.0010585784912109375
] |
[
0.9599609375,
0.0372314453125,
0.002079010009765625,
0.0006723403930664062
] |
The IM-UNITI trial reported higher rates of drug persistence in the 8-week arm versus q12w in CD, but a nonsignificant trend in corticosteroid-free remission .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277193_p87
|
PMC11277193
|
sec[3]/p[5]
|
4. Conclusions
| 2.8125 |
biomedical
|
Study
|
[
0.99365234375,
0.005023956298828125,
0.0015649795532226562
] |
[
0.78759765625,
0.1796875,
0.0301971435546875,
0.0027523040771484375
] |
Some studies evaluated the further shortening of UST administration intervals to 6 or 4 weeks, or a second IV reinduction in patients with unsatisfactory response to “conventional” dose escalation.
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277193_p88
|
PMC11277193
|
sec[3]/p[6]
|
4. Conclusions
| 2.310547 |
biomedical
|
Study
|
[
0.99462890625,
0.002593994140625,
0.00299072265625
] |
[
0.82763671875,
0.148681640625,
0.0222320556640625,
0.0013933181762695312
] |
Similarly, the UNIFI trial also favors q8w versus q12w administration in UC . Due to the more recent approval by regulatory institutions, data on 6- and 4-week administration of UST in UC are less extensive. Nonetheless, data confirm similar rates to those reported in CD.
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277193_p89
|
PMC11277193
|
sec[3]/p[7]
|
4. Conclusions
| 3.994141 |
biomedical
|
Study
|
[
0.9990234375,
0.0008440017700195312,
0.0003483295440673828
] |
[
0.5166015625,
0.004795074462890625,
0.477783203125,
0.0006799697875976562
] |
Considering the low immunogenicity of UST compared to anti-TNF agents , and the annual loss of response rate being 21% per patient/year in primary responders , the likelihood of effective dose escalation in UST is high. Response is recaptured following dose escalation in approximately 50% of CD and UC patients who lost response , thus comparable to those observed with TNFα antagonists and vedolizumab. Some evidence supports the efficacy of UST IV reinduction or IV administration of the drug , but is still unclear whether the intravenous route of administration offers advantage over short-interval SC doses.
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277193_p90
|
PMC11277193
|
sec[3]/p[8]
|
4. Conclusions
| 3.986328 |
biomedical
|
Study
|
[
0.9990234375,
0.0005893707275390625,
0.00020933151245117188
] |
[
0.99072265625,
0.0020160675048828125,
0.00696563720703125,
0.0001558065414428711
] |
UST concentrations exceeding 3.3 µg/mL or 4.5 µg/mL following induction are associated with better results . High UST trough levels are associated with more favorable results and are significantly higher in patients treated with q8w than q12w, more so following dose escalation by reducing dose intervals to q6w or q4w . This is expected and in keeping with the findings in patients treated with anti-TNFα and biologics interfering with leukocyte trafficking .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277193_p91
|
PMC11277193
|
sec[3]/p[9]
|
4. Conclusions
| 3.957031 |
biomedical
|
Review
|
[
0.9951171875,
0.002857208251953125,
0.0017843246459960938
] |
[
0.13818359375,
0.035552978515625,
0.82568359375,
0.0007143020629882812
] |
Long-term data on safety and drug persistence following UST dose escalation are lacking, but, all given, this is not cause of concern. Instead, the positioning of UST reinduction or dose escalation, instead of switching out of class, following loss of response strongly depends on cost evaluation. UST is indeed more expensive than traditional immunosuppressants and anti-TNFα agents, but comparable to other more recent biologics. Cost coverage by individual national health systems is variable in differing countries, as well as timing and extent of repricing politics. This, and safety considerations, represent the major factors in decision making, more so considering that the mean age of patients treated in the real-world experience is higher than that in clinical trials .
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277193_p92
|
PMC11277193
|
sec[3]/p[10]
|
4. Conclusions
| 4.09375 |
biomedical
|
Review
|
[
0.98583984375,
0.0101470947265625,
0.004077911376953125
] |
[
0.00649261474609375,
0.001438140869140625,
0.99169921875,
0.0005779266357421875
] |
In conclusion, according to registrative trials and real-world evidence, UST is safe and effective in IBD. However, like all other biologics, in addition to primary nonresponders, a proportion of patients lose response. The overall annual risk of loss of response is 21% per patient/year in CD and that of dose escalation in primary responders is 25% per patient/year . Data reported in the present narrative review indicate that over half of these patients may benefit from dose escalation or reinduction and recapture response without switching out of class. This evidence is supported by a number of observational and cohort studies, including a large series of patients, although it is still unclear which patients will most likely profit from more aggressive therapeutic strategies. A formal evaluation of the efficacy of UST reinduction or dose escalation, however, was not the primary outcome of randomized clinical trials. This prevented recommending this approach in the most recent international guidelines , and represents the main limitation of the present review. Another limitation resides in the use of data so far published as Congress abstracts. All considered, the off-label use of UST in the real-world experience is more frequent than expected and it seems effective and safe. UST risk stratification trials, as well as head-to-head comparisons with other biologics and small molecules, are needed to fully exploit the potential of this drug in a patient-tailored approach.
|
[
"Filippo Vernia",
"Sabrina Monaco",
"Giovanni Latella"
] |
https://doi.org/10.3390/jcm13143993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p0
|
PMC11277205
|
sec[0]/p[0]
|
1. Introduction
| 3.978516 |
biomedical
|
Review
|
[
0.9921875,
0.006328582763671875,
0.0016803741455078125
] |
[
0.0645751953125,
0.1729736328125,
0.75927734375,
0.0031833648681640625
] |
The prevalence of non-specific functional and somatoform physical complaints in primary care is stated as a proportion between 15 and 31 percent . Such patients typically present to the general practitioner’s surgery with a wide variety of physical complaints without sufficient anatomical findings . During the course of a long medical history, patients affected often have an exaggerated physical perception and show signs of psychological dysfunction . Despite the lack of indications of a cause and repeated assurances by doctors that the signs of illness are not physically justifiable, patients affected are convinced of a physical cause. In comparison, the possibility of a psychological problem as the trigger of the complaints is usually dismissed . As a matter of fact, apart from biological or genetic predispositions, there can be drastic and fundamental problems behind somatoform disorders, e.g., drastic life events, chronic diseases or serious illness experiences, social conflicts, lack of support, occupational stress, extreme body observation, health anxiety or hypochondria .
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p1
|
PMC11277205
|
sec[0]/p[1]
|
1. Introduction
| 3.816406 |
biomedical
|
Other
|
[
0.85986328125,
0.1312255859375,
0.0090179443359375
] |
[
0.014373779296875,
0.9736328125,
0.007785797119140625,
0.0039825439453125
] |
In the context of somatoform disorders, there are several problems involved in doctor–patient interaction . Firstly, patients with unclear physical complaints often behave in an appellative manner that is determined by a persistent insistence on specific examinations (e.g., further diagnostics) . Secondly, they meet with an understanding of their medical history from doctors, and a procedure that is intended to guarantee that no hidden illness is overlooked and that all potential trigger factors are included . Thirdly, if anatomical causes cannot be found, there may be a discrepancy between the respective assumptions of the cause, which bears additional conflict potential . If the doctor distinctly rules out anatomical causes and considers other potential explanations, the patient might feel misunderstood, accuse the doctor of professional incompetence and terminate the doctor–patient relationship .
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p2
|
PMC11277205
|
sec[0]/p[2]
|
1. Introduction
| 2.933594 |
biomedical
|
Other
|
[
0.97412109375,
0.0186920166015625,
0.00717926025390625
] |
[
0.0017595291137695312,
0.9833984375,
0.01331329345703125,
0.0014190673828125
] |
An interdisciplinary S3 guideline on the clinical picture of somatoform disorders exists for German-speaking countries , whose recommendations for diagnostic and therapeutic issues are generally also suitable for the field of general practitioners. Furthermore, diagnostic instruments exist (e.g., International Diagnosis Checklist for ICD-10 ‘Somatoform Disorder’ or PHQ-15 for the assessment of the severity of somatic symptoms) . With ICD-11 and even more so DSM-5, the concept of somatoform disorders gradually changes. What is now important is not so much that the physical complaints have no explainable pathology, but rather that they are stressful for the patient.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277205_p3
|
PMC11277205
|
sec[0]/p[3]
|
1. Introduction
| 3.394531 |
biomedical
|
Study
|
[
0.97509765625,
0.0170135498046875,
0.00792694091796875
] |
[
0.4287109375,
0.383056640625,
0.18359375,
0.004474639892578125
] |
While research projects on aspects such as prevalence and helpful therapeutic strategies have already been submitted for the German-speaking countries, there is a lack of studies that focus how general practitioners actually deal with these patients in daily practice . Individual studies have been able to show that unspecific physical complaints without organic findings are widespread in general practitioners’ practices, and that this number has risen continuously in recent years. Unclear symptoms are often associated with pronounced health anxiety. In their everyday practice, which are short on time and resources, general practitioners are faced with the challenge of clarifying these often-complex symptoms and setting the course for appropriate further care . In particular, communication strategies for establishing a stable doctor–patient relationship, which forms the basis for successful management, have hardly been researched to date.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p4
|
PMC11277205
|
sec[0]/sec[0]/p[0]
|
Research Interest
| 3.751953 |
biomedical
|
Study
|
[
0.9833984375,
0.014373779296875,
0.00202178955078125
] |
[
0.99560546875,
0.0016813278198242188,
0.001972198486328125,
0.00061798095703125
] |
The aim of the study is to investigate how general practitioners recognise the symptoms of somatoform disorders in daily healthcare situations, what significance they attach to them and how they deal with affected patients. A special focus is placed on the determination of experienced patient characteristics, suspected causes of somatoform disorders, challenges experienced in consultation, care and stabilisation strategies, as well as diagnostic forms of support.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p5
|
PMC11277205
|
sec[1]/sec[0]/p[0]
|
2.1. Study Design and Survey Instrument
| 1.987305 |
biomedical
|
Study
|
[
0.96533203125,
0.0020751953125,
0.032562255859375
] |
[
0.9638671875,
0.034759521484375,
0.0009264945983886719,
0.0005841255187988281
] |
Quantitative survey: The explorative study was designed as an online survey, by means of which general practitioners were questioned. On the one hand, it is based on a comprehensive literature search. On the other hand, previous studies by the authors involving health anxiety were taken into consideration (e.g., ).
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p6
|
PMC11277205
|
sec[1]/sec[0]/p[1]
|
2.1. Study Design and Survey Instrument
| 2.429688 |
biomedical
|
Study
|
[
0.97998046875,
0.0149993896484375,
0.005031585693359375
] |
[
0.6572265625,
0.337646484375,
0.0025882720947265625,
0.002655029296875
] |
The designed survey instrument (see Supplementary Material , completion time: approx. 12 min) consists of the following components: (a) occurrence of somatoform disorders in daily practice; (b) patient characteristics; (c) diagnostic procedure and guideline orientation; (d) challenges experienced and stabilisation strategies; (e) interdisciplinary cooperation; (f) optimisation proposals.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p7
|
PMC11277205
|
sec[1]/sec[0]/p[2]
|
2.1. Study Design and Survey Instrument
| 1.833008 |
biomedical
|
Study
|
[
0.96630859375,
0.01309967041015625,
0.0206451416015625
] |
[
0.98046875,
0.0175933837890625,
0.0008821487426757812,
0.0010633468627929688
] |
In addition to the standardised questions, several open questions were included (6, 8, 16, 21). The socio-demographic features collected were gender, age, specialist background, practice environment, type of practice and number of patients per quarter. A pre-test was conducted with 15 randomly selected general practitioners prior to use in the field.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p8
|
PMC11277205
|
sec[1]/sec[0]/p[3]
|
2.1. Study Design and Survey Instrument
| 1.874023 |
biomedical
|
Other
|
[
0.876953125,
0.035552978515625,
0.0875244140625
] |
[
0.2685546875,
0.72412109375,
0.00395965576171875,
0.0034656524658203125
] |
Qualitative interviews: The supplementary guideline for the semi-standardised interviews with general practitioners was deliberately based on the quantitative survey, although open questions were formulated here in accordance with the qualitative paradigm. In addition, the focus of the short interviews is on preferred strategies for the management of somatoform disorders.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p9
|
PMC11277205
|
sec[1]/sec[1]/p[0]
|
2.2. Recruitment and Participants
| 1.806641 |
biomedical
|
Study
|
[
0.76708984375,
0.0308074951171875,
0.2021484375
] |
[
0.7763671875,
0.2203369140625,
0.0009489059448242188,
0.00222015380859375
] |
Quantitative survey: Between January and August 2023, all 13,170 general practitioners actively treating patients in Hesse , Rhineland-Palatinate and Baden-Württemberg received a written invitation to participate in the anonymised survey. The survey was a one-off letter in which the doctors to be surveyed were given password-protected access to the online survey. The participants did not receive any incentives.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277205_p10
|
PMC11277205
|
sec[1]/sec[1]/p[1]
|
2.2. Recruitment and Participants
| 3.048828 |
biomedical
|
Study
|
[
0.951171875,
0.0182037353515625,
0.030609130859375
] |
[
0.99560546875,
0.00386810302734375,
0.00021696090698242188,
0.00031304359436035156
] |
Qualitative interviews: The recruitment of general practitioners for the qualitative interviews differed from that of the quantitative survey. First, a pool of 515 contact addresses was prepared with the help of the federal state-specific Internet doctor finder of the Association of Statutory Health Insurance Physicians, which contained a wide range of general practitioner practices in all federal states (30 to 35 addresses per federal state). For the German non-city states, it was ensured that all administrative districts were represented, that individual and joint practices occurred in approximately equal numbers, and that different practice environments were represented. Each federal state had to be equally represented in the study, regardless of the number of its inhabitants, in order to better represent rural regions in particular. When selecting the number of four general practitioners envisaged for each federal state, the following access criteria applied: at least 1 joint practice, balanced gender relationship, balanced relationship between urban and rural-small town practices, and consideration of older and younger doctors. A total of 82 general practitioner surgeries throughout Germany were contacted based on the deliberations described above. If interest in study participation was signalled, the doctors received study information. The recruitment period extended from December 2022 to March 2023. A total of 64 general practitioners were finally recruited for individual interviews.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p11
|
PMC11277205
|
sec[1]/sec[1]/p[2]
|
2.2. Recruitment and Participants
| 1.514648 |
other
|
Other
|
[
0.322509765625,
0.00234222412109375,
0.6748046875
] |
[
0.473876953125,
0.5224609375,
0.0023174285888671875,
0.0013284683227539062
] |
All 64 interviews were conducted alternately by the authors between March and April 2023 in person orally, by telephone or via instant messaging service (30 to 45 min). The interviews were recorded.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p12
|
PMC11277205
|
sec[1]/sec[2]/p[0]
|
2.3. Data Analysis
| 3.673828 |
biomedical
|
Study
|
[
0.998046875,
0.00023055076599121094,
0.0016450881958007812
] |
[
0.9951171875,
0.004779815673828125,
0.00024890899658203125,
0.00007081031799316406
] |
Quantitative survey: After cleansing the data set, the data were analysed using SPSS 23.0 for Windows. In order to determine significant differences between two groups, a t-test was used for independent samples (mean difference at the p < 0.001 level). This parametric method has a high-test strength and is considered to be statistically robust. The necessary conditions were met with the number of cases, the normal distribution of the groups to be distinguished and the fact that the samples come from the same population .
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p13
|
PMC11277205
|
sec[1]/sec[2]/p[1]
|
2.3. Data Analysis
| 1.766602 |
other
|
Other
|
[
0.325439453125,
0.0016841888427734375,
0.6728515625
] |
[
0.11700439453125,
0.87890625,
0.003170013427734375,
0.0008630752563476562
] |
The evaluation of the open questions is based on post-coding in the sense of the qualitative content analysis .
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p14
|
PMC11277205
|
sec[1]/sec[2]/p[2]
|
2.3. Data Analysis
| 2.236328 |
other
|
Study
|
[
0.4033203125,
0.0012378692626953125,
0.595703125
] |
[
0.72216796875,
0.275634765625,
0.0014028549194335938,
0.0006031990051269531
] |
Qualitative interviews: The first indications of theoretical saturation resulted after 48 interviews. The transcripts prepared after collection of the data were evaluated by the lead author with the help of a qualitative content analysis according to Mayring (MAXQDA). The category system generated was closely oriented to the guideline and was repeatedly checked and, if necessary, modified as the evaluation continued. The pseudonymized codification of the interviews was carried out as follows: e.g., I-55-m (number of the interview; gender).
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p15
|
PMC11277205
|
sec[2]/sec[0]/p[0]
|
3.1. Sample
| 2.121094 |
biomedical
|
Study
|
[
0.9541015625,
0.01119232177734375,
0.034637451171875
] |
[
0.98779296875,
0.0115509033203125,
0.0003287792205810547,
0.00042057037353515625
] |
Of the 2839 questionnaires processed, 2797 fully completed forms were included in the evaluation (response rate: 21%). Based on the socio-demographic characteristics collected, the sample is structured as follows: Gender: 53% male, 47% female; Average age: 54 (median: 53); Practice environment: 53% medium-sized and large cities, 47% rural-small towns; Type of practice: 52% individual practices, 44% joint practices, 4% other; Number of patients per quarter: 18% < 1000, 37% 1000–1500, 22% 1501–2000, 23% > 2000.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p16
|
PMC11277205
|
sec[2]/sec[0]/p[1]
|
3.1. Sample
| 1.842773 |
biomedical
|
Study
|
[
0.7529296875,
0.00859832763671875,
0.23828125
] |
[
0.86083984375,
0.137451171875,
0.0008273124694824219,
0.0009541511535644531
] |
The sample obtained for the qualitative interviews is composed of: Gender: 22 male, 22 female; Average age: 55 (median: 55); Practice environment: 44 medium- and large-sized cities, 20 rural-small towns; Type of practice: 38 individual practices, 26 joint practices; Status: 48 practice owners, 16 employed doctors.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p17
|
PMC11277205
|
sec[2]/sec[1]/p[0]
|
3.2. Somatoform Disorders in Everyday Practice
| 1.865234 |
biomedical
|
Study
|
[
0.9189453125,
0.043487548828125,
0.037689208984375
] |
[
0.87548828125,
0.1181640625,
0.0033626556396484375,
0.0027446746826171875
] |
When asked about the assumed proportion of non-specific somatoform complaints in practice consultations, 43% stated a range between 16 and 20%, followed by 26% who assumed 10–15%.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p18
|
PMC11277205
|
sec[2]/sec[1]/p[1]
|
3.2. Somatoform Disorders in Everyday Practice
| 1.714844 |
biomedical
|
Study
|
[
0.85986328125,
0.039215087890625,
0.100830078125
] |
[
0.54833984375,
0.444091796875,
0.003955841064453125,
0.0035457611083984375
] |
Thirty-nine percent of all respondents assumed that the number of patients affected by non-specific somatoform physical complaints has been seen to increase in general practitioner care in recent years. Twenty-six percent stated that the proportion had increased moderately (twenty-five percent: not increased).
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p19
|
PMC11277205
|
sec[2]/sec[2]/p[0]
|
3.3. Patient Characteristics
| 2.347656 |
biomedical
|
Study
|
[
0.98388671875,
0.01140594482421875,
0.004756927490234375
] |
[
0.953125,
0.04351806640625,
0.0021800994873046875,
0.0011577606201171875
] |
With regard to various symptoms, 85% of respondents reported the occurrence of panic attacks and anxiety disorders among affected patients, followed by persistent pain such as headaches, chest or back pain (84%). In the experience of 72%, patients complained of fatigue and exhaustion, but also of cardiovascular (64%) and gastrointestinal (59%) complaints.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p20
|
PMC11277205
|
sec[2]/sec[2]/p[1]
|
3.3. Patient Characteristics
| 2.513672 |
biomedical
|
Study
|
[
0.8779296875,
0.10040283203125,
0.021514892578125
] |
[
0.943359375,
0.05169677734375,
0.002239227294921875,
0.002681732177734375
] |
In patients with somatoform problems, the respondents observed an increased frequency of consultation in connection with numerous questions about their own complaints, as well as the demand for instrument-based diagnostics (cf. Table 1 ). The respondents also frequently stated false or exaggerated expectations regarding the services to be provided by the general practitioner. Doctors often noticed an extreme sensitivity and nervousness in patients. Once patients were convinced that they know the cause of their illness, the respondents experienced difficulty in distracting them from their opinions and concerns.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p21
|
PMC11277205
|
sec[2]/sec[2]/p[2]
|
3.3. Patient Characteristics
| 2.130859 |
biomedical
|
Study
|
[
0.9775390625,
0.005802154541015625,
0.0164947509765625
] |
[
0.87548828125,
0.1209716796875,
0.002208709716796875,
0.0011959075927734375
] |
When it comes to suspected or actual causes for the development of somatoform disorders, the respondents primarily stated extraordinarily stressful situations, such as unemployment, separation, accidents, operations or the loss of a close person (85%), but also stress and conflicts at work (80%). A routinely anxious way of handling physical complaints or hypochondria (71%) was also mentioned.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p22
|
PMC11277205
|
sec[2]/sec[3]/p[0]
|
3.4. Diagnostic Procedure and Guideline Orientation
| 2.033203 |
biomedical
|
Study
|
[
0.9658203125,
0.0088958740234375,
0.025177001953125
] |
[
0.73681640625,
0.259521484375,
0.0019702911376953125,
0.0017261505126953125
] |
At 30%, a smaller proportion coded in the area of neurotic stress and somatoform disorders as per ICD. Of these, 21% in turn stated that they do so frequently, and another 28% occasionally. Another part of the respondents stated that they confine themselves to the description of comorbidities such as depression, anxiety disorders and/or diagnoses such as unclassified chronic pain.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p23
|
PMC11277205
|
sec[2]/sec[3]/p[1]
|
3.4. Diagnostic Procedure and Guideline Orientation
| 1.618164 |
other
|
Other
|
[
0.372314453125,
0.2420654296875,
0.385498046875
] |
[
0.015533447265625,
0.9765625,
0.0025119781494140625,
0.005184173583984375
] |
In an enquiry, reasons were identified as to why the ICD code is hardly used for this clinical picture. At the centre of this is the fact that the general practitioners questioned considered an early determination of the ICD code to be premature and little differentiated in view of such a complex ailment. “The ICD terminology is too specific and requires a quick determination. Can I be sure that it is not in fact depression or some other problem?” “It is often difficult to be sure what is actually behind such complaints and how they might interact with each other. So, I avoid ICD.”
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p24
|
PMC11277205
|
sec[2]/sec[3]/p[2]
|
3.4. Diagnostic Procedure and Guideline Orientation
| 1.404297 |
other
|
Other
|
[
0.321533203125,
0.0102691650390625,
0.66845703125
] |
[
0.02520751953125,
0.97265625,
0.0011167526245117188,
0.0011434555053710938
] |
In this regard, there is too little knowledge of the content of somatoform physical complaints, so that a determination of the ICD code seems rushed. Many respondents did not claim to make a specific diagnosis but concentrated on excluding common physical causes to a large extent. “For me, my work ends where I find no indication of a clear physical cause.”
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p25
|
PMC11277205
|
sec[2]/sec[3]/p[3]
|
3.4. Diagnostic Procedure and Guideline Orientation
| 1.530273 |
biomedical
|
Other
|
[
0.60302734375,
0.0268096923828125,
0.369873046875
] |
[
0.0118408203125,
0.9853515625,
0.001708984375,
0.0010280609130859375
] |
Other respondents state the lacking consistency of coding. “Nothing is achieved by categorising the patient.” “This is all about holistic medicine, i.e., looking at the patients in all their complexity and helping them as well as possible. An ICD code is only a label. ”
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p26
|
PMC11277205
|
sec[2]/sec[3]/p[4]
|
3.4. Diagnostic Procedure and Guideline Orientation
| 2.177734 |
biomedical
|
Study
|
[
0.95654296875,
0.0245361328125,
0.018798828125
] |
[
0.62255859375,
0.3583984375,
0.01496124267578125,
0.004039764404296875
] |
A follow-up question revealed that only 15% of the doctors questioned the use of certain diagnostic tools such as (psychosomatic) checklists or forms for diagnostics or for differential diagnostic deliberations for this clinical picture. An analogous picture emerges with regard to the interdisciplinary S3 guideline on somatoform disorders. Twenty-eight percent of the respondents stated that they are aware of this.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p27
|
PMC11277205
|
sec[2]/sec[3]/p[5]
|
3.4. Diagnostic Procedure and Guideline Orientation
| 1.688477 |
biomedical
|
Other
|
[
0.74951171875,
0.134765625,
0.115478515625
] |
[
0.276123046875,
0.703125,
0.0128326416015625,
0.0077667236328125
] |
Thirty-nine percent of the doctors familiar with the S3 guideline assessed it as being quite useful. In an open question, numerous respondents criticised the guideline: “ inconvenient and impractical ”, “ too distant from application ”, “ no individual consideration of the patient possible ”, “ does not do justice to the complexity of psychosomatic disorders ”.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p28
|
PMC11277205
|
sec[2]/sec[4]/p[0]
|
3.5. Challenges and Stabilisation Strategies
| 2.542969 |
biomedical
|
Study
|
[
0.97265625,
0.0142974853515625,
0.01322174072265625
] |
[
0.9912109375,
0.007808685302734375,
0.0007333755493164062,
0.0004146099090576172
] |
Sixty-five percent of all respondents experienced the management of patients with non-specific somatoform physical complaints as very or rather strenuous in their daily practice (twenty-seven percent less or not at all strenuous). The proportion of doctors who generally experience challenges was clearly higher among doctors in small towns and rural communities than in medium- and large-sized cities (76% to 54%, p < 0.001).
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p29
|
PMC11277205
|
sec[2]/sec[4]/p[1]
|
3.5. Challenges and Stabilisation Strategies
| 2.246094 |
biomedical
|
Study
|
[
0.76123046875,
0.14013671875,
0.098876953125
] |
[
0.68798828125,
0.302978515625,
0.0052947998046875,
0.003971099853515625
] |
With regard to dealing with somatoform patients, the doctors questioned experienced the most varied challenges (cf. Table 2 ). In addition to the problem of providing sufficient time for consultation, a clear majority of the respondents find it rather difficult to influence these patients in such a way that excessive fear of the presence of a serious illness can be prevented and misunderstandings avoided. Closely linked to this is ensuring compliance. About three quarters of those questioned find it challenging to motivate patients to make use of appropriate psychosocial support.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p30
|
PMC11277205
|
sec[2]/sec[4]/p[2]
|
3.5. Challenges and Stabilisation Strategies
| 2.179688 |
biomedical
|
Study
|
[
0.7099609375,
0.2313232421875,
0.058990478515625
] |
[
0.75,
0.2364501953125,
0.006988525390625,
0.006656646728515625
] |
Despite the requirements and problems involved with somatoform complaints, the respondents state numerous strategies with which they have had positive experiences in stabilising patients (cf. Table 3 ). Most of the respondents articulated that they consider in-depth and continuous intercommunication with the patient in a relaxed conversation situation to be particularly important to enable the development of a trusting doctor–patient interaction at eye level. A large part of the doctors attached importance to assessing the patient’s background and life situation.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p31
|
PMC11277205
|
sec[2]/sec[4]/p[3]
|
3.5. Challenges and Stabilisation Strategies
| 2.332031 |
clinical
|
Other
|
[
0.43408203125,
0.51220703125,
0.053924560546875
] |
[
0.0033130645751953125,
0.9912109375,
0.0013837814331054688,
0.004245758056640625
] |
The restraint exercised during tangential discussion is intended to avoid raising false expectations, provide rational clarification and avoid excessive examinations. This may possibly require an early relativisation of inappropriate wishes and requests. Regarding their approach, the respondents attempt to address psychosocial topics rather casually at first. In the opinion of the respondents, regular appointments, i.e., appointments that are independent of complaints and fears, should be arranged for a limited period of time. In order to introduce patients to the clinical picture of somatoform disorders, information material is sometimes compiled. Complaints diaries are provided to help determine more precisely under which everyday conditions the symptoms occur.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p32
|
PMC11277205
|
sec[2]/sec[4]/p[4]
|
3.5. Challenges and Stabilisation Strategies
| 1.905273 |
biomedical
|
Other
|
[
0.91943359375,
0.016448974609375,
0.06414794921875
] |
[
0.08538818359375,
0.9091796875,
0.0037078857421875,
0.0019006729125976562
] |
Apart from the reference to relaxation techniques, several doctors emphasised the importance of regular physical activity as a therapeutic instrument, e.g., to change awareness routines and support the retrieval of subjectively perceived sovereignty in daily life. Most of the respondents were clearly hesitant to treat somatoform disorders with medication.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p33
|
PMC11277205
|
sec[2]/sec[5]/p[0]
|
3.6. Qualitative Findings
| 2.839844 |
clinical
|
Other
|
[
0.2406005859375,
0.72216796875,
0.0369873046875
] |
[
0.0157318115234375,
0.97216796875,
0.0041656494140625,
0.00806427001953125
] |
In the interviews, most of the general practitioners interviewed also made it clear that they attach particular importance to an in-depth and continuous interaction with the patient in a dialogue situation that is as relaxed as possible. In their opinion, a relaxed, supportive and empathetic attitude, as well as a patient-centred discussion, are of paramount importance for a trusting doctor–patient interaction. When obtaining information, the doctor should ensure to treat the somatoform patient as a partner at eye level, i.e., listen to his/her complaints in detail and address them without reservations. For some doctors, it is important to ask questions at an early stage to help them assess the patient’s background, life situation and personality (e.g., high level of body awareness, hypersensitivity), as this may be relevant for further management and the assessment of therapeutic options.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p34
|
PMC11277205
|
sec[2]/sec[5]/p[1]
|
3.6. Qualitative Findings
| 2.419922 |
clinical
|
Other
|
[
0.291259765625,
0.603515625,
0.10498046875
] |
[
0.043670654296875,
0.9462890625,
0.0032062530517578125,
0.006610870361328125
] |
The restraint exercised during tangential discussion is intended to avoid raising false expectations, provide rational clarification and avoid excessive examinations. This may possibly require an early relativisation of inappropriate wishes and requests, but in a way that the patient himself/herself is more likely to realise which expectations are realistic and which are not. With regard to their approach, the doctors interviewed reported that they attempt to address psychosocial issues in a casual and indirect way, using terms from daily life. They considered this a critical point, because if patients have the feeling of being “pushed into a psychological category” at an early stage, they may feel that their concerns are not being taken seriously. The following is, therefore, important:
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11277205_p35
|
PMC11277205
|
sec[2]/sec[5]/p[2]
|
3.6. Qualitative Findings
| 1.587891 |
other
|
Other
|
[
0.28955078125,
0.319580078125,
0.390869140625
] |
[
0.0103912353515625,
0.98486328125,
0.0014495849609375,
0.0033588409423828125
] |
In the opinion of the respondents, regular appointments, i.e., appointments that are independent of complaints and fears, should be arranged for a limited period of time. Apart from realistic and possibly verifiable discussion and therapy targets, some of the doctors emphasised that one should “not do too much in too short a time”. This leads to a tense situation for both doctor and patient.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p36
|
PMC11277205
|
sec[2]/sec[5]/p[3]
|
3.6. Qualitative Findings
| 2.011719 |
biomedical
|
Other
|
[
0.861328125,
0.09051513671875,
0.04833984375
] |
[
0.01010894775390625,
0.98486328125,
0.0030117034912109375,
0.0019350051879882812
] |
Regarding specific measures for the management of somatoform patients, some of the respondents first of all emphasise the importance of continuously working on the “awareness of the patients […] that there are symptoms that occur without a clear physical cause”. In some cases, information material is compiled for this purpose. Complaints diaries are intended to help isolate symptoms and determine more precisely under which everyday conditions these symptoms occur.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p37
|
PMC11277205
|
sec[2]/sec[5]/p[4]
|
3.6. Qualitative Findings
| 1.701172 |
biomedical
|
Other
|
[
0.85986328125,
0.0220794677734375,
0.11822509765625
] |
[
0.0024089813232421875,
0.994140625,
0.002376556396484375,
0.0009465217590332031
] |
Apart from referring to relaxation techniques, several doctors emphasised the importance of regular physical activity as a therapeutic instrument. This is not just about moving awareness routines, but about supporting the retrieval of subjectively perceived sovereignty and a sense of control in daily life.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p38
|
PMC11277205
|
sec[2]/sec[5]/p[5]
|
3.6. Qualitative Findings
| 1.55957 |
other
|
Other
|
[
0.4091796875,
0.12103271484375,
0.4697265625
] |
[
0.0026721954345703125,
0.99560546875,
0.0006880760192871094,
0.0010166168212890625
] |
In the course of the development of compact online therapy offered by certain health insurance companies, some general practitioners are trying to introduce somatoform patients to such offers where psychosocial care appears to be necessary. The idea is that patients often resist standard therapies but may possibly accept a low-threshold, anonymous offer of help.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p39
|
PMC11277205
|
sec[2]/sec[5]/p[6]
|
3.6. Qualitative Findings
| 2.845703 |
biomedical
|
Other
|
[
0.84326171875,
0.14599609375,
0.01085662841796875
] |
[
0.037109375,
0.9501953125,
0.008697509765625,
0.004238128662109375
] |
Most of the interviewed general practitioners usually avoided a medication-based treatment of somatoform disorders. Only in individual cases, in which psychopharmacological interventions exist, for example due to simultaneous depression, anxiety, obsessive–compulsive or sleep disorders, are attempts made to include medication-based solutions. Here, it is preferably attempted to treat occurring pain, sleep disorders and depressive symptoms simultaneously, if possible, with a single medication. In this context, however, the respondents insist on a functioning and closely scheduled interdisciplinary cooperation.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p40
|
PMC11277205
|
sec[2]/sec[6]/p[0]
|
3.7. Interdisciplinary Cooperation
| 3.322266 |
biomedical
|
Study
|
[
0.982421875,
0.01166534423828125,
0.006038665771484375
] |
[
0.99365234375,
0.004520416259765625,
0.0015430450439453125,
0.0003426074981689453
] |
Regarding interdisciplinary cooperation, only some of the respondents had the impression that they can rely on cooperation with regional physicians for diagnostic clarification and/or treatment of somatoform complaints. Forty percent felt that the cooperation in this area is very good or rather good, whereas fifty percent considered it as rather poor or very poor. While 55% of the physicians in large- and medium-sized cities assessed the cooperation with colleagues from psychiatry/psychosomatics and neurology as good, only 26% of physicians in small towns and rural communities were of the same opinion ( p < 0.001). A similar picture emerges with regard to cooperation with psychotherapists. Forty-two percent considered this as very good or rather good, while fifty-one percent rated it as rather poor or very poor.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p41
|
PMC11277205
|
sec[2]/sec[6]/p[1]
|
3.7. Interdisciplinary Cooperation
| 2.654297 |
biomedical
|
Study
|
[
0.91845703125,
0.0675048828125,
0.01403045654296875
] |
[
0.9775390625,
0.01898193359375,
0.0017185211181640625,
0.0016880035400390625
] |
A follow-up question provided information on the causes of the dissatisfaction established. Thus, 41% considered that the specialist colleagues in psychiatry and psychotherapy usually provide sufficient information about the examinations, results and/or therapeutic measures performed. In the case of psychotherapists, 65% stated that due to long waiting times, they have become rather reluctant as regards referrals and rather seek other solutions, if necessary. Seventy-five percent stated that they often feel left alone in the management of patients with somatoform complaints. While the proportion among urban doctors was 61%, it was 88% among rural doctors ( p < 0.001).
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p42
|
PMC11277205
|
sec[2]/sec[7]/p[0]
|
3.8. Optimisation Suggestions
| 1.708008 |
biomedical
|
Other
|
[
0.630859375,
0.01922607421875,
0.35009765625
] |
[
0.10150146484375,
0.89501953125,
0.002201080322265625,
0.0015306472778320312
] |
With a view to the potential optimisation of the care of somatoform physical complaints, some of the respondents (47%) named the creation of significantly more psychotherapeutic therapy places. This was followed by various suggestions intended to anchor health insurance companies more strongly as contact persons for psychosomatic problems (35%).
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277205_p43
|
PMC11277205
|
sec[2]/sec[7]/p[1]
|
3.8. Optimisation Suggestions
| 1.834961 |
biomedical
|
Other
|
[
0.68212890625,
0.024444580078125,
0.293212890625
] |
[
0.03662109375,
0.95947265625,
0.0028743743896484375,
0.0009331703186035156
] |
The availability of online therapy services, whether provided by certain health insurance companies or in the form of digital health applications, was considered to be promising. Some of the respondents (32%) attempt to introduce somatoform patients to such services where psychosocial care appears to be necessary. The idea is that patients often resist standard therapies but may possibly accept a low-threshold, anonymous offer of help.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277205_p44
|
PMC11277205
|
sec[3]/sec[0]/p[0]
|
4.1. Main Findings and Comparison with Previous Studies
| 2.425781 |
biomedical
|
Study
|
[
0.96630859375,
0.0185699462890625,
0.0148773193359375
] |
[
0.8330078125,
0.158447265625,
0.0059051513671875,
0.002574920654296875
] |
The findings of the quantitative and qualitative survey show that in the perception of general practitioners, somatoform disorders make up a considerable and increasing proportion of patient consultation. Patients with somatoform disorders have a characteristic behavioural profile that may pose challenges for general practitioners . Therefore, patients affected can trigger reservations, negative positioning and conflictual attitudes in general practitioners . In light of the findings, it is evident that general practitioners bear potential psychological causes and intensifying stress factors in mind.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p45
|
PMC11277205
|
sec[3]/sec[0]/p[1]
|
4.1. Main Findings and Comparison with Previous Studies
| 3.953125 |
biomedical
|
Study
|
[
0.986328125,
0.01042938232421875,
0.0034198760986328125
] |
[
0.97314453125,
0.003582000732421875,
0.0225372314453125,
0.0006427764892578125
] |
The quantitative and qualitative findings are closely related and can be considered jointly in almost every respect. The results show that general practitioners consider a stable doctor–patient relationship to be a central prerequisite for successful treatment. Therefore, they make various efforts to establish a resilient, tangential basis for consultation . This is accompanied by measures to consistently exclude physical causes on the one hand and to enable the best possible assessment of patients, as well as to gently introduce them to the clinical picture of somatoform disorders, on the other. The general practitioners questioned mainly attach importance to providing sufficient consultation time for this group of patients. It is worth noting that in English-language studies, such as Wortman et al., similar points are raised, including ‘Continuously searching for common ground’ and ‘Making patients aware of the interaction between body and mind’ . As Stone already observed, general practitioners may experience the interaction with patients affected by physically unexplained complaints as personally and ethically enhancing, as the image of holistic, speaking medicine can fully develop here . In addition, the approaches selected show parallels to the treatment of patients with health anxiety, who are associated with the phenomenon of cyberchondria .
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p46
|
PMC11277205
|
sec[3]/sec[0]/p[2]
|
4.1. Main Findings and Comparison with Previous Studies
| 2.056641 |
biomedical
|
Study
|
[
0.96484375,
0.009063720703125,
0.026123046875
] |
[
0.54443359375,
0.3583984375,
0.09234619140625,
0.00475311279296875
] |
Knowledge and application of the S3 Guidelines was surprisingly low; there were reservations and criticism among the respondents as regards their applicability. A major problem seems to be a lack of practical relevance and over-complexity . In the paper by Natschke et al., the attitude of general practitioners with regard to the S3 guideline on somatoform complaints was also critical . The authors then proposed the preparation of a user-friendly version for general practitioners.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p47
|
PMC11277205
|
sec[3]/sec[0]/p[3]
|
4.1. Main Findings and Comparison with Previous Studies
| 3.914063 |
biomedical
|
Study
|
[
0.990234375,
0.00720977783203125,
0.002445220947265625
] |
[
0.9072265625,
0.03350830078125,
0.05804443359375,
0.0011043548583984375
] |
However, it seems questionable whether such a practice-oriented, compact version will change the basic finding that general practitioners tend to avoid diagnostic guidelines and tools for somatoform disorders and proceed according to their personal understanding of the case in question . An explicit diagnosis as a somatoform disorder is obviously not experienced as helpful in dealing with the symptoms, similar to depression . This indicates that general practitioners experience a diagnosis as a potential problem in dealing with patients, and that the background of the occurrence cannot be clarified due to the individual differences of somatoform problems . Earlier studies were also able to prove that questionnaires, for example, are only used to a limited extent as screening instruments in GP surgeries . Coding according to ICD, therefore, can be described as challenging, as numerous overlapping diagnoses are described .
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p48
|
PMC11277205
|
sec[3]/sec[0]/p[4]
|
4.1. Main Findings and Comparison with Previous Studies
| 3.832031 |
biomedical
|
Study
|
[
0.9609375,
0.0245361328125,
0.01446533203125
] |
[
0.80908203125,
0.14111328125,
0.047821044921875,
0.0019359588623046875
] |
General practitioners experience various difficulties at the interfaces of interdisciplinary cooperation with colleagues from psychiatry/psychosomatics and neurology, as well as with psychotherapists. Long waiting times and communication problems (e.g., lack of direct case-related exchange) mean that general practitioners can often only rely on themselves. Consistently noteworthy in the findings of this study is that rural doctors experience the care of somatoform disorders as a greater challenge. This may be related to the fact that doctors outside urban centres can rely far less on an adequate care and support network. This shows the necessity for better cooperation and the strengthening of interdisciplinary structures . In the opinion of the doctors questioned, low-threshold solutions, such as online therapies, can help to compensate for the lack of psychosomatic care capacities in somatoform disorders. Here, the statutory health insurance companies in particular can play a more proactive role . The development of new offers, such as digital health apps, represents a major opportunity in this context.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11277205_p49
|
PMC11277205
|
sec[3]/sec[1]/p[0]
|
4.2. Strengths and Limitations
| 1.919922 |
biomedical
|
Study
|
[
0.96435546875,
0.003902435302734375,
0.031646728515625
] |
[
0.9599609375,
0.038726806640625,
0.0010156631469726562,
0.0005116462707519531
] |
Although it was possible to obtain a large, heterogeneous sample of primary care providers, various limitations of the survey must be reflected. These include a regional recruitment focus in three federal states and a limited response (quantitative survey). Moreover, it is possible that general practitioners with an interest in the subject may have shown stronger participation in the survey (selection bias).
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11277205_p50
|
PMC11277205
|
sec[3]/sec[1]/p[1]
|
4.2. Strengths and Limitations
| 2.150391 |
biomedical
|
Study
|
[
0.98974609375,
0.0007042884826660156,
0.00933074951171875
] |
[
0.97021484375,
0.026458740234375,
0.0030727386474609375,
0.0004527568817138672
] |
This study was only a cross-sectional study and only correlative relationships can be uncovered but—due to the lack of longitudinal data—not robust causal relationships.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277205_p51
|
PMC11277205
|
sec[4]/p[0]
|
5. Conclusions
| 3.175781 |
biomedical
|
Other
|
[
0.76171875,
0.216796875,
0.02130126953125
] |
[
0.26953125,
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0.03631591796875,
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The results show that general practitioners consider a good and stable doctor–patient relationship as a central basis for successful treatment. General practitioners apply a wide range of communication and stabilisation strategies in the care of somatoform physical complaints. Therefore, they demonstrate competence and important prerequisites for mastering the communicative challenges that are to be expected in the treatment of somatoform disorders. Nevertheless, contact with this group of patients is experienced as a particular challenge in daily practice. In the long run, apart from the creation of more therapeutic care capacities and interdisciplinary structures, it would be helpful to strengthen low-threshold therapy and support services.
|
[
"Julian Wangler",
"Michael Jansky"
] |
https://doi.org/10.3390/ijerph21070901
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p0
|
PMC11277217
|
sec[0]/p[0]
|
1. Introduction
| 4.085938 |
biomedical
|
Study
|
[
0.99951171875,
0.00021731853485107422,
0.00013685226440429688
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[
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Osteochondroma is the most common benign bone tumor, accounting for 20–50% of benign bone tumors in children . Patients may develop solitary osteochondromas due to injuries at the metaphysis or multiple osteochondromas associated with mutations in the Exostosin 1 ( EXT1 ) and 2 ( EXT2 ) genes, a condition known as hereditary multiple osteochondromas (HMO; previously called hereditary multiple exostoses, HME) . Osteochondromas are not present at birth in patients with HMO. However, approximately 96% of affected individuals develop multiple osteochondromas by 12 years of age .
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p1
|
PMC11277217
|
sec[0]/p[1]
|
1. Introduction
| 3.759766 |
biomedical
|
Other
|
[
0.99609375,
0.0035152435302734375,
0.0005497932434082031
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[
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In most cases, osteochondromas are diagnosed using radiography or computed tomography (CT) . Patients with HMOs currently rely on surgery for treatment. Patients undergo an average of 2.7 surgeries in their lifetime for osteochondromas . However, repetitive surgeries pose a severe burden on patients, families, and the healthcare system. Depending on the location of the osteochondroma, the surgical procedures can vary from simple to complex. If the entire cartilaginous cap is not removed, osteochondromas can recur or malignant transformation may occur . Preventing surgery will minimize recovery time and healthcare costs and reduce the risk of complications.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277217_p2
|
PMC11277217
|
sec[0]/p[2]
|
1. Introduction
| 4.042969 |
biomedical
|
Study
|
[
0.9990234375,
0.0008640289306640625,
0.00018799304962158203
] |
[
0.91943359375,
0.00872039794921875,
0.0712890625,
0.0007677078247070312
] |
To date, three drugs have been demonstrated to inhibit osteochondroma formation in mouse models of multiple osteochondroma (MO) models. They are LDN-193189, a bone morphogenetic protein antagonist , PF-04449913, an inhibitor of Indian hedgehog , and palovarotene, a specific agonist of retinoic acid nuclear receptor gamma (RARγ) . However, no Food and Drug Administration (FDA)-approved drugs are currently available for the treatment of osteochondromas. A clinical phase 2 trial was performed to investigate the efficacy and safety of palovarotene for the treatment of MO. However, a clinical hold for this trial was issued following safety reports of early growth plate closure in pediatric patients observed during another clinical trial for a lethal genetic disease, fibrodisplasia ossificance progressiva (FOP) . Although the FDA has approved palovarotene as a treatment for patients with FOP , studies on the clinical application of palovarotene for MO have remained limited.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999994 |
PMC11277217_p3
|
PMC11277217
|
sec[0]/p[3]
|
1. Introduction
| 4.144531 |
biomedical
|
Study
|
[
0.99951171875,
0.00040841102600097656,
0.0001914501190185547
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[
0.99951171875,
0.00024819374084472656,
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0.00008153915405273438
] |
This study was performed to examine the application of palovarotene and other RARγ agonists for the treatment of osteochondromas. The study has three aims. One is to determine whether palovarotene inhibits the growth of pre-existing osteochondromas because patients generally visit clinicians after osteochondroma formation is detected. The second aim is to determine whether local application of the RARγ agonist can inhibit osteochondroma growth, minimizing systemic adverse effects. The third aim is to investigate alterations of the molecular pathways in palovarotene-treated osteochondroma chondrocytes because elucidation of molecular pathways affected by the RARγ agonist should be valuable in developing alternative drugs for osteochondromas. We observed that palovarotene halted the growth of pre-existing osteochondromas in increasing doses with age. Furthermore, local application of the RARγ agonists inhibited osteochondroma growth for a limited period of time. In addition, we demonstrated that RARγ agonists stimulated the signal transducer and activator of the transcription 3 (Stat3) signaling pathway in osteochondroma cells.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277217_p4
|
PMC11277217
|
sec[1]/sec[0]/p[0]
|
2.1. Measurement of Cartilage Tumor Volume by Enhanced microCT in an AcanCreER;Ext1f/f Mouse Osteochondroma Model
| 4.308594 |
biomedical
|
Study
|
[
0.99951171875,
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0.9990234375,
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] |
Various mouse osteochondroma models have been reported . We chose the AcanCreER;Ext1f/f compound transgenic mouse model generated by crossing AcanCreER mice with Ext1 e2neofl mice. In this transgenic mouse line, tamoxifen-dependent Cre recombination can be induced in neonates and at older ages. To confirm the efficiency and selectivity of Cre recombination in AcanCreER mice, tamoxifen was injected into AcanCreER;R26R ZsGreen mice starting at the neonatal (P5) and infant (P17) stages. In both cases, Cre recombination in the growth plate was efficiently and selectively induced three days after tamoxifen injection, as evidenced by the expression of ZsGreen reporter proteins . When tamoxifen was injected at P5 and P7, ectopic cartilage masses, referred to as osteochondromas, were observed in the periphery of the proximal growth plate in the ulna and radius in AcanCreER;Ext1f/f mice but not in Ext1f/f mice 2.5 weeks after the injections, indicating that tumor formation after tamoxifen injections is dependent on the AcanCreER transgene. However, the induction of ectopic cartilage was much less prominent when tamoxifen was injected into P17 . Similar differences in the degree of osteochondroma induction were observed at the costochondral junctions of the ribs . These results indicate that the effective induction of osteochondromas in the AcanCreER;Extf/f mouse model requires Cre recombination at the neonatal stage. Sex-based differences were observed in the degree of osteochondroma formation . In this study, we induced osteochondromas in AcanCreER;Extf/f male mice, starting at P5.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277217_p5
|
PMC11277217
|
sec[1]/sec[0]/p[1]
|
2.1. Measurement of Cartilage Tumor Volume by Enhanced microCT in an AcanCreER;Ext1f/f Mouse Osteochondroma Model
| 4.121094 |
biomedical
|
Study
|
[
0.99951171875,
0.00028133392333984375,
0.00022995471954345703
] |
[
0.99951171875,
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One pitfall in the analysis of osteochondromas is the difficulty of accurately determining their volume. Previous studies have used histomorphometric analysis and the number of ectopic cartilage masses to evaluate tumor formation . In this study, we used a phosphotungstic acid (PTA)-enhanced microCT. Without PTA staining, microCT only recognized the bone, as shown in the transverse and coronal planes . After PTA staining, reconstructed microCT images showed both bone and cartilaginous osteochondromas located in the ulna and radius of the distal forelimb . We could distinguish the perichondrium from the osteochondromas . The osteochondroma region was manually defined, and the entire volume of the selected region was determined. Increases in the volume of tumors from 3.5 weeks of age (2.5 weeks after tamoxifen injection) to 7.5 weeks of age (6.5 weeks after tamoxifen injection) were evident .
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p6
|
PMC11277217
|
sec[1]/sec[1]/p[0]
|
2.2. Inhibition of Pre-Existing Osteochondromas by Palovarotene
| 4.132813 |
biomedical
|
Study
|
[
0.99951171875,
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[
0.99951171875,
0.0003192424774169922,
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0.00008153915405273438
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To determine whether palovarotene inhibited the growth of pre-existing osteochondromas, AcanCreER;Ext1f/f mice received tamoxifen injections at P5 and P7 and daily palovarotene gavage starting at 3.5 weeks of age when osteochondromas were visible . A 2-week treatment with 1.76 mg/kg palovarotene strongly reduced tumor growth compared with the control group, whereas a 2-week treatment with 0.26 mg/kg palovarotene did not significantly reduce tumor growth . Treatment with 1.76 mg/kg palovarotene inhibited tumor growth to a size similar to that at the beginning of treatment . When treatment with 1.76 mg/kg palovarotene was prolonged for an additional 2 weeks, osteochondromas were evident in both the ulna and radius of the forelimbs , although the tumor volume was smaller than that in the control group . To determine whether the weakening of antitumor action was rescued by increasing the dose, we changed the regimen to 1.76 mg/kg for 2 weeks, followed by 4 mg/kg for 2 weeks. This regimen strongly reduced the osteochondroma volume in both the ulna and radius of the forelimbs . We did not observe a significant difference in tibia length between control and palovarotene-treated AcanCreER;Ext1f/f mice . However, the palovarotene regimen resulted in a reduction in tibia length in Ext1f/f mice compared to that in the vehicle control .
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p7
|
PMC11277217
|
sec[1]/sec[1]/p[1]
|
2.2. Inhibition of Pre-Existing Osteochondromas by Palovarotene
| 4.109375 |
biomedical
|
Study
|
[
0.99951171875,
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0.00018155574798583984
] |
[
0.99951171875,
0.0002541542053222656,
0.00027298927307128906,
0.00006651878356933594
] |
Given the in vivo data indicating that palovarotene injection reduced the tibia length in mice, we sought to determine whether the local injection of RARγ agonists can ameliorate skeletal toxicity. AcanCreER;Ext1f/f mice received tamoxifen injections at P5 and P7 and received an injection of nanoparticles containing 1 μg of the NRX204647 RARγ agonist (NRX-NP) using a micro-injector. The injections were delivered twice a week for 2 weeks in the right distal wrist. PTA-enhanced microCT analysis demonstrated that local injection of NRX-NPs inhibited osteochondroma growth . However, extending the treatment for an additional two weeks with the same regimen failed to suppress tumor growth . NRX-NP treatment did not cause an imbalance in limb length in either AcanCreER;Ext1f/f or Ext1f/f mice, as determined by measurement of the ulna length .
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p8
|
PMC11277217
|
sec[1]/sec[2]/p[0]
|
2.3. Actions of Palovarotene on Osteochondromas
| 4.386719 |
biomedical
|
Study
|
[
0.99951171875,
0.00040841102600097656,
0.0002009868621826172
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[
0.9990234375,
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Palovarotene stimulates cartilage matrix degradation and inhibits cartilage matrix synthesis in human osteochondromas in explant cultures . To determine whether the inhibition of osteochondroma growth by palovarotene in mice was associated with these inductive changes, osteochondroma sections were prepared from tamoxifen-injected AcanCreER;Ext1f/f mice one week after palovarotene or vehicle treatment. These two groups showed different histology. Vehicle-treated control osteochondroma showed a growth plate-like structure rotated at a 90° angle . In contrast, palovarotene-treated osteochondroma tissues were divided into proximal and distal parts containing more hypertrophied cells than in the control group . The proximal portion of the palovarotene-treated osteochondroma was composed of smaller cells. Immunohistochemical staining with an antibody recognizing the C-terminal neoepitope of aggrecanase-cleaved aggrecan (NITEGE) revealed that the palovarotene-treated osteochondromas contained larger amounts of degraded aggrecan products than in the control group . The in situ hybridization demonstrated the upregulation of matrix metalloproteinase 13 and a disintegrin and metalloproteinase with thrombospondin motifs 5 . The expression of SRY-box transcription factor 9 ( Sox9 ) was downregulated in the palovarotene-treated osteochondromas . Inhibition of cell proliferation activity by palovarotene was not evident, as determined using the immunohistochemistry of Ki67 . These results indicate that palovarotene stimulates matrix degradation in osteochondroma tissues, which may play an important role in inhibiting tumor growth.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999994 |
PMC11277217_p9
|
PMC11277217
|
sec[1]/sec[3]/p[0]
|
2.4. Actions of Palovarotene on Cultured Chondrocytes
| 4.355469 |
biomedical
|
Study
|
[
0.99951171875,
0.00038886070251464844,
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[
0.9990234375,
0.00028443336486816406,
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0.00011336803436279297
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To understand the direct action of palovarotene on osteochondroma cells, we performed transcriptome analysis of cultured chondrocytes . AcanCreER;Ext1f/f and Ext1f/f littermates received tamoxifen injections at P5 and P7, and epiphyseal chondrocytes were isolated from these mice at P10. The isolated cells were embedded in alginate beads, acclimated to culture for 2 days, and treated with 300 nM palovarotene or vehicle ethanol for 2 days. The cells were then released from the alginate gels and subjected to bulk RNA-seq. Differentially expressed genes (DEGs) were identified by both false discovery rate (FDR) <0.05 and log2 fold change with cutoff values of ±1 . A comparison between palovarotene-treated and control AcanCreER;Ext1f/f chondrocytes revealed that 2582 genes were significantly upregulated and 2222 genes were downregulated. A comparison between palovarotene-treated and control Ext1f/f chondrocytes demonstrated that 2629 and 2450 genes were significantly upregulated and downregulated, respectively . A comparison between AcanCreER;Ext1f/f and Ext1f/f chondrocytes and between palovarotene-treated AcanCreER;Ext1f/f and Ext1f/f chondrocytes revealed that a small number of genes were significantly altered, with 33 genes upregulated and 27 genes downregulated in AcanCreER;Ext1f/f chondrocytes compared to those in Ext1f/f chondrocytes, and 49 genes upregulated in palovarotene-treated AcanCreER;Ext1f/f chondrocytes compared to those in Ext1f/f chondrocytes. Ext1 expression was significantly reduced in AcanCreER;Ext1f/f and palovarotene-treated AcanCreER;Ext1f/f . Since full completion of Cre recombination in Ext1 e2neofl mice is expected to reduce Ext1 expression by 50% , the efficiency of recombination in AcanCreER;Ext1f/f chondrocytes by tamoxifen injections was considered to be 30–50%. Functional analysis of the DEGs was performed using Ingenuity Pathway Analysis (IPA) . The DEGs in the palovarotene-treated AcanCreER;Ext1f/f chondrocytes compared to those in control AcanCreER;Ext1f/f chondrocytes were strongly linked to the upregulation of the hypoxia-inducible factor 1a (HIF1a) signaling and osteoarthritis pathways . Canonical pathway analysis demonstrated that the DEGs in AcanCreER;Ext1f/f chondrocytes compared to Ext1f/f chondrocytes were highly linked to the downregulation of the osteoarthritis pathway.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277217_p10
|
PMC11277217
|
sec[1]/sec[3]/p[1]
|
2.4. Actions of Palovarotene on Cultured Chondrocytes
| 4.191406 |
biomedical
|
Study
|
[
0.99951171875,
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[
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Comparison analysis of two conditions ( AcanCreER;Ext1f/f vs. Ext1f/f and palovarotene-treated AcanCreER;Ext1f/f vs. control AcanCreER;Ext1f/f ) revealed that multiple pathways, including the osteoarthritis pathway, were altered in the opposite manner , suggesting that the genes linked to these biological pathways are important for both osteochondroma formation and palovarotene-induced inhibition of osteochondroma growth. The possible upstream regulators that drive the counteracting changes between Ext1 deficiency and palovarotene treatment included retinoid drugs (tretinoin), inflammatory molecules (lipopolysaccharide, interferon-gamma [IFNG], and interleukin 1B [IL1B]), and metabolic regulators (3,5-dihydroxyphenylglycine and insulin-induced gene 1 [INSIG1]) . We performed a comparison analysis of the results obtained from palovarotene-treated AcanCreER;Ext1f/f cultured chondrocytes and palovarotene-treated human osteochondroma explants . Activation of the interferon signaling pathway was observed in both palovarotene-treated cultures .
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p11
|
PMC11277217
|
sec[1]/sec[4]/p[0]
|
2.5. Stimulation of the Stat3 Pathway by Palovarotene
| 4.152344 |
biomedical
|
Study
|
[
0.99951171875,
0.000232696533203125,
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[
0.99951171875,
0.00021517276763916016,
0.00023818016052246094,
0.00005835294723510742
] |
The link between palovarotene action and the interferon signaling pathway was further analyzed. Expressions of the upregulated genes listed in the interferon signaling pathway were examined in palovarotene-treated osteochondromas. The expression of cytochrome P450 family 26 subfamily member 1 ( Cyp26b1 ), a genomic target molecule of the retinoid signaling pathway, was upregulated in the osteochondroma region in the periphery of the growth plate of palovarotene-treated AcanCreER;Ext1f/f mice , indicating that osteochondromas responds to palovarotene. Expression of Ifnar1 was detected in osteochondromas , and palovarotene treatment did not upregulate the expression level . In contrast, the expression of Ifngr1 was upregulated in palovarotene-treated osteochondromas .
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p12
|
PMC11277217
|
sec[1]/sec[4]/p[1]
|
2.5. Stimulation of the Stat3 Pathway by Palovarotene
| 4.367188 |
biomedical
|
Study
|
[
0.99951171875,
0.0003399848937988281,
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] |
[
0.9990234375,
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0.00010341405868530273
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Interferon signaling is mediated by the phosphorylation of Stat1 (signal transducer and activator of transcription 1) and/or Stat2 . Cultured chondrocytes were treated with palovarotene or mouse recombinant interferon beta protein (mIFNb) for 2 and 24 h, and cell lysates were subjected to immunoblotting. Slight stimulation of the phosphorylation of Stat1 and Stat2 was detected 2 h after the IFNb treatment, but not after 24 h. Palovarotene did not induce phosphorylation of Stat1 or Stat2 at either time point . Phosphorylation of Stat3 is stimulated by the type I interferon signaling pathway . Phosphorylation of Stat3 was strongly stimulated by palovarotene and IFNb 24 h after treatment , suggesting that palovarotene stimulated the Stat3 pathway. Selective activation of the Stat3 pathway, but not Stat1 or Stat2, was confirmed by the results of a reporter assay using the Cignal Pathfinder. The Stat3 pathway was significantly stimulated , while the Stat1 and Stat1/2 pathways were not . Palovarotene increased the STAT3 reporter activity in RARγ-hetero (RARγ+/−) chondrocytes, but not in RARγ-null chondrocytes , indicating that Stat3 activation was specific to RARγ. Furthermore, palovarotene-treated osteochondromas contained more phospho-Stat3 . These results indicate that palovarotene stimulates the Stat3 pathway.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277217_p13
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PMC11277217
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sec[2]/sec[0]/p[0]
|
3.1. Clinically Relevant Regimen of Drug Treatment for Osteochondromas
| 4.066406 |
biomedical
|
Study
|
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Patients generally present to the clinic with pre-existing osteochondromas after they notice lumps made by growing cartilaginous tumors. In contrast, preclinical studies on osteochondroma treatment have used a regimen in which drug treatment begins at the onset or very early stage of tumor formation . In this study, palovarotene treatment was initiated after osteochondromas developed to a certain level . This treatment stopped the increase in tumor volume , suggesting that palovarotene has an anti-tumor effect on pre-existing osteochondromas. The doses of palovarotene orally administrated in mice, 1.76 mg/kg and 4.0 mg/kg correspond to human equivalent doses of 0.14 mg/kg and 0.32 mg/kg, respectively, according to the practice guide on dose conversion between mice and humans. These converted doses are comparable to the flare-up doses of palovarotene (SOHONOS) for patients with FOP .
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p14
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sec[2]/sec[0]/p[1]
|
3.1. Clinically Relevant Regimen of Drug Treatment for Osteochondromas
| 4.375 |
biomedical
|
Study
|
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Palovarotene treatment did not result in complete tumor regression, and the dose of palovarotene had to be increased to achieve complete tumor suppression over time . Histological analyses indicated that the mechanism of tumor inhibition involved the destruction of cartilaginous tissue by degradation of the cartilage matrix and stimulation of chondrocyte hypertrophy rather than by the suppression of the proliferation of osteochondroma cells . The findings indicate that combined treatment with anti-proliferative drugs may have better therapeutic effects. To date, no serious adverse events, including early growth plate closure, have been reported in a clinical study of multiple osteochondromas (MO) . However, this clinical trial was placed on hold because of the toxicities observed with chronic dosing in the FOP trial . Because the aforementioned MO study was primarily a pediatric study with an upper age of enrollment of 14 years, a period of life when skeletal growth is active, this discontinuation was unavoidable. We observed that increasing the dose of palovarotene required to maintain the long-term suppression of tumor growth negatively affected skeletal growth in mice , suggesting that high doses and long-term administration of the RARγ agonist in this mouse model have adverse effects on skeletal cells such as chondrocytes and osteoblasts. Site-specific delivery in combination with controlled drug release could be beneficial by enhancing therapeutic effects and reducing the toxicity associated with systemic drug administration. Local drug delivery using nanocarriers has been shown to both improve efficacy and minimize side effects on non-target tissues . Nanocarrier-based formulations can not only deliver drugs to their local sites of action but also protect labile compounds from premature degradation, increasing and sustaining drug bioavailability. As an experimental approach for suppressing the localized disease while minimizing systemic skeletal toxicity, we tested the nanoparticle-based local delivery of a RARγ agonist. NRX-NP significantly inhibited tumor growth , with our results demonstrating the feasibility of nanoparticle-based drug delivery, potentially paving the way to the development of a safer and more effective, site-specific drug therapy for osteochondromas. However, the inhibition efficacy was less (59% inhibition) than that of the systemic administration (88% inhibition) in the 2-week treatment and faded in the 4-week treatment . Previously, we succeeded in inhibiting the growth of xenografts of well-differentiated human chondrosarcoma cells over 7 weeks using the same NRX-NP . The difference in the effectiveness of the NRX-NP in these two systems can be attributed to several factors. The tested human chondrosarcoma cell xenograft was more sensitive to RARγ agonists than that were mouse osteochondromas. The anatomical location of osteochondromas surrounding the bones makes it difficult to target tumor cells and retain nanoparticles for a long period. Finally, the location of the xenograft in the middle of the back made it easy to apply a large amount of NRX-NP. Improvements in drug delivery systems and application techniques will be the future direction of this project.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p15
|
PMC11277217
|
sec[2]/sec[1]/p[0]
|
3.2. Actions of Palovarotene on Chondrocytes
| 4.484375 |
biomedical
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RARγ plays an important role in the regulation of cartilage development . RARγ is dominantly expressed in cartilage compared to other RAR family members, RARα and RARβ. RARγ expression starts at the time of the appearance of cartilaginous skeletons in embryos . RARγ mostly exists as a ligand-free form and supports the expression and accumulation of cartilage matrix, including sulfated proteoglycans and type II collagens . Retinoic acid treatment of limb buds and chondrocytes strongly inhibits the synthesis of proteoglycans and type II and IX collagens . These inhibitory effects of retinoid signaling on chondrogenesis and cartilage matrix formation have been considered to be the mechanism of the antitumor action of palovarotene on osteochondromas; the inhibition of aberrant bone morphogenetic protein signaling by retinoid signaling has been addressed . The stimulation of retinoid signaling by deletion of Cyp26b1, a responsible enzyme for retinoic acid degradation, in the growth plate inhibits chondrocyte proliferation and skeletal growth , indicating that silencing of retinoid signaling, presumably via RARγ, is required for proliferation of the growth plate chondrocytes. Hence, the stimulation of retinoid signaling by palovarotene was expected to inhibit chondrocyte proliferation in osteochondromas. However, this effect was not observed in our system.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277217_p16
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sec[2]/sec[1]/p[1]
|
3.2. Actions of Palovarotene on Chondrocytes
| 4.515625 |
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Histological examination demonstrated that palovarotene stimulated the degradation of the cartilage matrix and inhibited the expression of Sox9 , a master gene for cartilage matrix synthesis, leading to the destruction of the developing cartilaginous mass. RNA-seq analysis of cultured chondrocytes revealed the biological pathways altered by palovarotene treatment . Among them, the upregulation of the osteoarthritis pathway is the most relevant explanation for our observations. The genes enriched in the osteoarthritis pathway include the upregulation of various matrix degradation enzymes, such as Adamts4 , Adamts5 , Mmp9 , and Mmp13 , and the downregulation of cartilage-related genes, such as collagen type II alpha 1 chain Col2a1 , frizzled-related protein ( Frzb ), Indian hedgehog ( Ihh ), and Sox9 ( Supplemental File , osteoarthritis pathway sheet). Changes in the expression of these molecules are believed to mediate disruption of the osteochondroma matrix. We observed that the osteoarthritis pathway was downregulated in chondrocytes isolated from tamoxifen-injected AcanCreER;Ext1f/f mice compared to those isolated from tamoxifen-injected Ext1f/f mice . Although the extent of deletion of the Ext1 gene was mild, downregulation of the osteoarthritis pathway may be responsible for the mechanism of osteochondroma formation in AcanCreER;Ext1f/f mice. The HIF1α signaling pathway plays an important role in cartilage development and is up-regulated in embryonic developing cartilage . Hypoxia stimulates HIFα signaling and promotes the expression of the chondrogenic phenotype in culture . The finding that palovarotene upregulated the HIF1α signaling pathway in chondrocytes while decreased expression of Ext1 downregulated the HIF1α signaling pathway seems contradictory to the current understanding of this pathway in the regulation of cartilage development. However, the genes enriched in this pathway identified by IPA were related to matrix degradation (upregulation of Mmps ) and angiogenesis (upregulation of Vegfs ). These events have been caused by molecular changes during hypoxia-induced metastasis of tumor cells . In addition, chondrogenic genes were hardly upregulated in this pathway ( Supplemental File , HIF1a pathway sheet).
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277217_p17
|
PMC11277217
|
sec[2]/sec[1]/p[2]
|
3.2. Actions of Palovarotene on Chondrocytes
| 4.605469 |
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RNA-seq data analysis using IPA revealed the upregulation of the interferon signaling pathway in both palovarotene-treated mouse chondrocytes and palovarotene-treated human osteochondroma explants . The relationship between interferons and retinoids has been previously studied in cancer . The combination of interferons and retinoids potentiates antitumor action in certain types of cancer cells, such as leukemia, mammary carcinoma, and squamous carcinoma cells. Although the molecular basis for the synergistic antitumor action of interferons and retinoids is not completely understood, studies with anti-sense-based knockout genetic screens have revealed the importance of family genes associated with retinoic acid and interferon-induced cell death, which are termed genes associated with retinoid-interferon-induced mortality (GRIM) . However, in palovarotene-treated chondrocytes, the GRIM-1 ( Shq1 ), GRIM-12 ( Txnrd1 ), and GRIM-19 ( Ndufa13 ) genes, which have been studied among 12 genes , were not significantly upregulated. The results of in situ hybridization were not always consistent with the RNA-seq results; upregulation of ifngr1 was observed in the palovarotene-treated osteochondroma in vivo, whereas upregulation of ifnar1 was not . Palovarotene stimulated Stat3 phosphorylation at Y405 in chondrocytes but not Stat1 or Stat2 phosphorylation . The reporter assay demonstrated that the RARγ agonist stimulated the Stat3 pathway but not the Stat1 and Stat1/2 pathways . Stat1 and Stat 2 are considered dominant mediators of the type I interferon signaling pathway, but Stat 3 is not an indispensable regulator and is involved in the fine balance of interferon signaling as a negative or positive regulator . The association between retinoid signaling and interferon signaling observed by transcriptome analysis may be specific to the Stat 3 pathway but not via crosstalk with a typical interferon pathway.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277217_p18
|
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|
sec[2]/sec[1]/p[3]
|
3.2. Actions of Palovarotene on Chondrocytes
| 4.496094 |
biomedical
|
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Stat3 is recognized as an acute downstream intracellular effector of inflammatory cytokines and growth factors, including IL-6 and EGFs, and is involved in the regulation of DNA transcription . Activation of Stat3 is controlled by the phosphorylation of Tyr705 and Ser727 of its Src homology domain. The involvement of the Stat3 pathway in the regulation of skeletal tissue homeostasis has recently received attention . Liang et al. reported that upregulation of Stat3 phosphorylation (Tyr 705) was observed in osteoarthritic articular cartilage in human samples and a surgery-induced osteoarthritis mouse model. In this study, the antagonist of the retinoic acid receptor-related orphan receptor-alpha (RORα) inhibits the IL6-Stat3 pathway in cultured human chondrocytes, but it remains unclear if RORα can activate the Stat3 pathway. Many independent studies have demonstrated the involvement of Stat3 signaling activation in degenerative changes in articular cartilage in osteoarthritis . Actions of the Stat3 pathway in the context of osteoarthritis pathology include the stimulation of matrix degradation and cell death . The genes enriched in the osteoarthritis pathway revealed by IPA of the DEGs in the palovarotene-treated chondrocytes included various matrix catabolic genes ( Adamts4 , Adamts5 , Mmp9 , Mmp10 , Mmp12 , and Mmp13 ) and cell death-related genes ( Casp1 , Casp3 , and Casp6 ). Adamts4 , Mmp9 , Mmp13 , Casp3 , and Casp6 are target genes of Stat 3 . Taken together, our findings suggest that palovarotene stimulates the Stat3 signaling pathway and that the Stat3 pathway may underlie the antitumor action of palovarotene on osteochondromas, as this pathway is involved in cartilage degeneration in osteoarthritis.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p19
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sec[3]/sec[0]/p[0]
|
4.1. Animals
| 4.082031 |
biomedical
|
Study
|
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All procedures were conducted following the National Institutes of Health guidelines of the Institutional Animal Care and USE Committee of the University of Maryland, Baltimore . AggrecanCre ERT2 , Ext1 e2neofl , and Rosa26-CAG-loxP-stop-loxP-ZsGreen mice were obtained from Jackson Laboratory (Bar Harbor, MN). We crossed AggrecanCre ERT2 mice with R26R-ZsG and Ext1 e2neofl to create AggrecanCre ERT2 ;R26R ZsGreen ( AcanCre ERT2 ;R26R ZsGreen ) and AggrecanCre ERT2 ; Ext1 e2neofl ( AcanCreER;Ext1f/f ) mice. To achieve Cre recombination, 40 mg/kg body weight of tamoxifen dissolved in corn oil was intraperitoneally injected twice at P5 and P7. RARγ-deficient mice were kindly provided by Dr. Pierre Chambon (INSERM, Paris, France). Animals were housed in a University Laboratory Animal Resources supervised animal facility with a 12-h light/dark cycle in a temperature (22 ± 1 °C) and humidity (55 ± 5%) controlled room. The animals were provided with hygienic animal bedding, and all cages contained wood shavings, bedding, and cotton pads. The health status of each animal was monitored throughout the experiments by investigators, animal veterinary technicians, and veterinarians, according to institutional guidelines. The mice were free from viral, bacterial, and parasitic pathogens during the experimental period.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277217_p20
|
PMC11277217
|
sec[3]/sec[1]/p[0]
|
4.2. Drug Treatment
| 3.957031 |
biomedical
|
Study
|
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Atomax Chemicals Co., Ltd. (Shenzhou, China) prepared two selective RARγ agonists . The company synthesizes chemicals as described in their respective patent applications with over 98% purity. Once we received the products, their quality and biological activity were verified using nuclear magnetic resonance and mass spectrometry, and the reporter assay using the HEK293 cells that are stably integrated contained a firefly luciferase gene under the control of retinoic acid response elements along with the full-length human RARG gene (BPS Bioscience, San Diego, CA, USA). The drug powder was stored at −30 °C under desiccated conditions until use. Stock solutions were kept in an amber tube with argon gas and stored at −30 °C. The prepared working solution was sufficient for a set of experiments, dispensed in tubes for one-time use, and kept at −30 °C.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p21
|
PMC11277217
|
sec[3]/sec[1]/p[1]
|
4.2. Drug Treatment
| 4.070313 |
biomedical
|
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|
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AcanCreER;Ext1f/f , Ext1f/f , or AcanCre ERT2 ;R26R ZsGreen mice received intraperitoneal injections of tamoxifen in corn oil/10% ethanol (40 mg/kg) at ages P5 and P7. At 3.5 weeks post-injection, AcanCreER;Ext1f/f, and Ext1f/f mice were sedated and received daily oral gavage of palovarotene or vehicle (corn oil). The treatment periods were 3 days, 1 week, 2 weeks, and 4 weeks (n = 3–6). Doses were 0.26 or 1.76 mg/kg of palovarotene or vehicle for 2 or 4 weeks, and 1.76 of mg/kg palovarotene or vehicle for 2 weeks, followed by 4.0 mg/kg of palovarotene or vehicle for an additional 2 weeks. Because the manifestation of tumors was clearer in male mice than in female mice , male mice were used for the drug treatment experiments.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p22
|
PMC11277217
|
sec[3]/sec[1]/p[2]
|
4.2. Drug Treatment
| 4.167969 |
biomedical
|
Study
|
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For local treatment, based on the chemical structure and receptor binding selectivity, NRX204647 was chosen as the RARγ agonist to be administered in PLA-based NPs (NRX-NP). NP was formulated by modifying the emulsification-solvent evaporation method using serum albumin as a colloidal stabilizer . Briefly, NRX204647 (15 mg) was co-dissolved in 200 mg PLA (Mn = 19 kDa, polydispersity index = 1.8; Birmingham Labs, Birmingham, AL, USA) in chloroform (5 mL). The organic phase was emulsified by sonication in an aqueous solution containing 150 mg of human serum albumin (Octapharma AB, Stockholm, Sweden) dissolved in 10 mL of deionized water, and the organic solvent was removed using rotary evaporation under reduced pressure. NP was passed through a sterile 5.0-µm membrane, lyophilized with glucose (5% w / v ) as a cryoprotectant, stored at −80 °C, and reconstituted in deionized water before use. The drug loading of NP was determined spectrophotometrically following extraction in sec -butanol against a suitable calibration curve. Measurements of NP sizes were performed using dynamic light scattering. NRX-NP (0.5 μg/1 μL at each site) was subcutaneously injected at the medial and lateral sites of the vicinity of the proximal growth plate of the radius and ulna of the mice injected with tamoxifen (40 mg/kg) at P5 and P7 twice a week for 2 weeks and 4 weeks, beginning at 3.5 weeks of age. At each endpoint, the forelimbs were harvested and processed for X-ray, histological, microCT, or in situ hybridization analysis.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
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|
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|
sec[3]/sec[2]/p[0]
|
4.3. Histology and Immunohistochemical Staining
| 4.167969 |
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|
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|
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Tissues for histological examination were fixed with 4% paraformaldehyde (PFA) for 24 h at 4 °C. Samples were decalcified with 10% ethylenediaminetetraacetic acid at pH 7.2 after capturing radiographic images. The samples were processed for paraffin embedding. Sagittal sections of 5-μm in thickness were created throughout the project. Sections were stained with HE and safranin O. For immunohistochemical staining, the sections were baked for 1 h at 60 °C and then deparaffinized. Sections (n = 2–4) were incubated in 3% hydrogen peroxide in methanol for 10 min. Sections were then incubated with 400 IU/mL hyaluronidase for 30 min at 37 °C (NITEGE) or 10 mM sodium citrate buffer solution (pH 6.0) for 8 min at 80 °C (Ki67 and phosphor-Stat3), followed by blocking buffer, 5% bovine serum albumin, and 1% goat serum for 60 min at room temperature. The sections were then incubated with the primary antibody against aggrecan degradation products , Ki67 , or phospho-Stat3 overnight at 4 °C. Sections were then incubated with a 1:200 dilution of biotinylated anti-rabbit IgG , followed by color detection using Vectastain Elite ABC HRP reagent RTU and counterstaining with methyl green (Alfa Aeser, Haverhill, MA, USA). The results were observed and captured using a BZX700 microscope (Keyence, Itasca, IL, USA).
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p24
|
PMC11277217
|
sec[3]/sec[3]/p[0]
|
4.4. Tumor Volumetric Analysis by PTA-Enhanced microCT
| 4.140625 |
biomedical
|
Study
|
[
0.99951171875,
0.0003113746643066406,
0.0001615285873413086
] |
[
0.99853515625,
0.0008587837219238281,
0.0004260540008544922,
0.00010275840759277344
] |
To establish specific protocols for PTA-enhanced microCT that allow microlevel visualization of cartilage tissues, including the growth plate and osteochondroma, we adapted and expanded a previously described protocol . Samples were immersed in 2% KOH for 3–7 days after fixation with 4% PFA overnight. Under a dissection microscope, the distal forelimb was exposed by removing tendons and soft tissues. Forelimbs were then incubated in 0.7% PTA in 70% ethanol at room temperature for one week. Samples were placed in 70% ethanol without PTA and scanned using a Skyscan 1172 microCT scanner (Bruker, Billerica, MA, USA). Image acquisition at a resolution of 6.5 µm/pixel required 45 min (parameters: 0.2 mm Al filter using a 55 kV X-ray source, resolution at 2 K, step rotation of 0.51, and two average frames using 360° rotation). Images were reconstructed using NRecon software (version 1.7.4.6, Bruker). The cartilage tumor area was manually selected, and the tumor volume was analyzed.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277217_p25
|
PMC11277217
|
sec[3]/sec[4]/p[0]
|
4.5. In Situ Hybridization by RNAscope
| 4.109375 |
biomedical
|
Study
|
[
0.99951171875,
0.000202178955078125,
0.00016796588897705078
] |
[
0.9951171875,
0.004383087158203125,
0.0005311965942382812,
0.0001627206802368164
] |
The forelimbs were fixed with 2% formaldehyde, dehydrated by increasing the sucrose percentage to 30%, and then embedded in the optimal cutting temperature (O.C.T.) compound (Thermo Fisher Scientific, Waltham, MA, USA). Undecalcified frozen sections were mounted on Kawamoto’s film and subjected to in situ hybridization using the RNAscope ® Fluorescent Multiplex Kit (Advance Cell Diagnostic Inc., Newark, CA, USA). Predesigned single- or multiplex probes ( Cyp26b1 , Mmp13 , Adamts5 , Sox9 , Infar1 , Infgr1 , and Itf1 ) were obtained from the same company.
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999994 |
PMC11277217_p26
|
PMC11277217
|
sec[3]/sec[5]/p[0]
|
4.6. Chondrocyte Culture
| 4.136719 |
biomedical
|
Study
|
[
0.99951171875,
0.0002225637435913086,
0.00017333030700683594
] |
[
0.9990234375,
0.0004363059997558594,
0.0003333091735839844,
0.00006973743438720703
] |
Primary mouse chondrocytes were isolated from AggCreER;Ext1f/f and Ext1f/f mice at P8 after injections of tamoxifen (40 mg/kg) at P5 and P7, using a previously reported protocol . The isolated cells were embedded in 1.2% alginate (Cosmo Bio USA, Carlsbad, CA, USA) at a density of 0.5 × 10 6 /mL following the manufacturer’s protocol and cultured in high glucose DMEM containing 10% fetal bovine serum under a 3% O 2 atmosphere. On day 2, the cultures were treated with 300 nM palovarotene or an equal volume of vehicle ethanol for 2 days. The cells were then released from alginate by incubation with 0.055 M sodium citrate, 0.03 M EDTA, and 0.15 M NaCl pH 6.8 for 5 min at 4 °C and lysed in RLT buffer (Qiagen, Redwood City, CA, USA).
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277217_p27
|
PMC11277217
|
sec[3]/sec[6]/p[0]
|
4.7. Transcriptome Analysis
| 4.199219 |
biomedical
|
Study
|
[
0.99951171875,
0.0003268718719482422,
0.00019288063049316406
] |
[
0.998046875,
0.0015878677368164062,
0.0003883838653564453,
0.00012195110321044922
] |
Total RNA preparation, library preparation, and RNA-seq analysis were performed at the Maryland Genomics Institute for Genome Sciences, University of Maryland School of Medicine. Total RNA was extracted from the cultured chondrocytes using the RNeasy Mini Kit (Qiagen, Hilden, Germany). Strand-specific, dual-unique indexed libraries for sequencing on all Illumina platforms were made using the NEBNext ® Ultra™ II Directional RNA Library Prep Kit for Illumina ® (New England Biolabs, Ipswich, MA, USA). The manufacturer protocol was modified by diluting adapter 1:30 and using 3 μL of this dilution. Library size selection was performed using AMPure SPRI-select beads (Beckman Coulter Genomics, Danvers, MA, USA). Glycosylase digestion of the adapter and second strand was performed in the same reaction as the final amplification to avoid further cleanup. Sample input for this method was polyA-enriched mRNA using the NEBNext ® Poly(A) mRNA Magnetic Isolation Module, a bead-based enrichment kit. Libraries were assessed for concentration and fragment size using a DNA High Sensitivity Assay on a LabChip GX Touch (Perkin Elmer, Waltham, MA, USA). Library concentrations were assessed using qPCR by the KAPA Library Quantification Kit (Complete, Universal) (Kapa Biosystems, Woburn, MA, USA).
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p28
|
PMC11277217
|
sec[3]/sec[6]/p[1]
|
4.7. Transcriptome Analysis
| 4.152344 |
biomedical
|
Study
|
[
0.99951171875,
0.0002225637435913086,
0.0001653432846069336
] |
[
0.99853515625,
0.0008664131164550781,
0.0003190040588378906,
0.00009179115295410156
] |
RNAseq analysis was carried out by Maryland Genomics, Institute for Genome Sciences, UMSOM. Paired-end Illumina libraries were mapped to the mouse reference, Ensembl release GRCm39.109, using HiSat2 v2.1.0, using default mismatch parameters. The DESeq2 Bioconductor package (v1.5.24) was used to estimate dispersion, normalize read counts by library size to generate the counts per million for each gene, and determine differentially expressed genes between disease and control samples. Differentially expressed transcripts with a p -value ≤ 0.05 and log 2 fold change ≥ 1 were used for downstream analyses. Normalized read counts were used to compute the correlation between replicates for the same condition and compute the principal component analysis for all samples. Pathway analysis was performed using IPA software . The data were deposited in the Gene Expression Omnibus database .
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277217_p29
|
PMC11277217
|
sec[3]/sec[7]/p[0]
|
4.8. Immunoblot Protocol
| 4.148438 |
biomedical
|
Study
|
[
0.99951171875,
0.0002925395965576172,
0.00017881393432617188
] |
[
0.9990234375,
0.0006384849548339844,
0.0003638267517089844,
0.00008082389831542969
] |
Epiphyseal chondrocytes were isolated from P3-P7 C57BL/6J , plated on type I collagen-coated 60 mm dishes at a density of 2.0 × 10 6 cells/cm 2, and cultured in high glucose DMEM containing 10% fetal bovine serum. After the cells grew to confluency, the medium was removed and replaced by high glucose DMEM containing 2% charcoal-treated serum with 300 nM palovarotene, 100 nM NRX204647, and 150 ng/mL mouse recombinant interferon beta (PBL Assay Science, Piscataway, NJ, USA) or 0.1% ethanol. Twenty-four hours after treatment, the cultures were lysed in RIPA buffer containing protease inhibitors, and the supernatants were stored at −80 °C until use. Cell lysates were subjected to SDS-PAGE (4–12%, NuPage Bis-Tris, Thermo Fisher Scientific) followed by western blotting with antibodies against phospho-Stat1 , phospho-Stat2 , and phosphor-Stat3 (all from Cell Signaling Technology). The antibodies bound to the membrane were visualized using the VECTASTATIN Elite Universal PLUS kit, peroxidase (Vector Laboratories), and SuperSignal West Femto (Thermo Fisher Scientific). The membranes were reblotted with an α-tubulin antibody (Thermo Fisher Scientific).
|
[
"Sonia A. Garcia",
"Kimberly Wilson",
"Ningfeng Tang",
"Hongying Tian",
"Takeshi Oichi",
"Aruni T. Gunawardena",
"Michael Chorny",
"Ivan S. Alferiev",
"John E. Herzenberg",
"Vincent Y. Ng",
"Masahiro Iwamoto",
"Motomi Enomoto-Iwamoto"
] |
https://doi.org/10.3390/ijms25147610
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
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