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PMC11277025_p16
|
PMC11277025
|
sec[1]/p[7]
|
2. Platelets: Small Cells with Great Importance in Health and Disease
| 3.691406 |
biomedical
|
Other
|
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PLTs can interact with pathogens by expressing antimicrobial molecules. This helps them to kill pathogens, and they can also engulf and store them, facilitating interaction with immune cells . The ability of PLTs to produce several antimicrobial molecules emphasizes their vital role in the initial response to detected abnormalities. These small blood cells act as a bridge between innate and adaptive immunity.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p17
|
PMC11277025
|
sec[1]/p[8]
|
2. Platelets: Small Cells with Great Importance in Health and Disease
| 3.96875 |
biomedical
|
Study
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PLTs have long been believed to play a role in transfusion-related immunomodulation (TRIM), independent of leukocytes . In a mouse model of immune thrombocytopenia, PLT transfusion was found to help stabilize PLT counts . In other studies, Ki et al. showed that stored PLT concentrates affected myeloid dendritic cells in various infection models. PLT transfusion led to the different regulation of co-stimulatory molecules and cytokine production depending on the specific infection model . Although PLT transfusions are essential and beneficial, it is important to be aware of their potential to modulate the immune system in different ways, as this can also lead to adverse events .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
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PMC11277025_p18
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PMC11277025
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sec[2]/p[0]
|
3. Storage Conditions for Platelet Concentrates
| 3.923828 |
biomedical
|
Study
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Storage conditions play a key role in maintaining PLT functionality. In contrast to red blood cells, storage at low temperatures compromises the viability of stored PLTs as it alters the ability to shapeshift and activate under various conditions . The quality of the PC and its in vivo effectiveness are also strongly dependent on the type of storage medium used or the application of the pathogen inactivation (PI) system .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
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PMC11277025_p19
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PMC11277025
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|
3.1. Standard Storage Conditions
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The guidelines for PLT storage recommend a temperature between 20 and 24 °C. These limits came from a study performed 60 years ago, in which the effect of storage at different temperatures for different durations on the in vivo recovery and survival of PLTs was demonstrated . The standard temperature for PLT storage in blood banks is 22 ± 2 °C with constant gentle horizontal agitation. This procedure allows storing PCs for up to 5 days in a closed system. This time could be extended to 7 days if appropriate packs and PC combinations were used. Due to concerns of bacterial contamination, the use of 7-day PCs requires an assay to exclude bacterial contamination prior to transfusion or application of a pathogen inactivation procedure . If any manufacturing step includes PLTs collecting in an open system, the PC should be used immediately after collection. Sometimes, storage is unavoidable. The PC should then be stored at a standard temperature of 22 ± 2 °C with continuous agitation and used within 6 h .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
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PMC11277025_p20
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PMC11277025
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sec[2]/sec[0]/p[1]
|
3.1. Standard Storage Conditions
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During the storage procedure, the PLTs should be gently agitated. If some interruptions appear (e.g., the integrity of the hermetic seal is broken) due to transportation, equipment failure, processing, etc., PCs are still suitable for use; therefore, the total time of interruption cannot be longer than 24 h, and a single interruption should not last longer than 8 h. If the conditions are fulfilled, the components are suitable for use with an unchanged shelf life . According to the most recent studies, constant agitation is still unavoidable. The shear forces induced by agitation keep the GPVI receptor down-regulated, attenuating PLT-spreading capacities during storage .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p21
|
PMC11277025
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sec[2]/sec[0]/p[2]
|
3.1. Standard Storage Conditions
| 4.101563 |
biomedical
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The effects of PLT storage at low temperatures (less than 20 °C) were demonstrated in two key papers. Gottschall and Aster stored PLTs at 18 °C, 19.5 °C, and 21 °C for 72 h. They found a significant effect on the PLTs’ mean life span, which was reduced from 8.1 days at 21 °C to 5.2 and 1.9 days after storage at 19.5 °C and 18 °C, respectively. These results clearly showed the need for quality control of temperature in PLT storage. Even if in vitro markers of PLT quality indicate a positive impact of temperatures below 20 °C, the most important is post-transfusion survival in vivo. In another study, PLTs were stored for 5 days at room temperature with or without 17 h of exposure to 16 °C or 12 °C. Post-transfusion recovery was slightly reduced after exposure to 16 °C or 12 °C (from 49% in the control group to 43% and 38%, respectively) . However, according to Gottschall’s research, PLT post-transfusion survival was also reduced from 6.5 to 3.5 and 2.0 days after exposure to 16 °C or 12 °C, respectively .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p22
|
PMC11277025
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sec[2]/sec[0]/p[3]
|
3.1. Standard Storage Conditions
| 2.845703 |
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Study
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Table 2 outlines the drawbacks associated with the current approaches to storing PLTs under standard conditions, along with proposed strategies for enhancing this storage process.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p23
|
PMC11277025
|
sec[2]/sec[1]/p[0]
|
3.2. Cold Storage
| 4.589844 |
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The effects of PLT storage at low temperatures were demonstrated in many articles . Currently, the temperature studied most often for cold storage is 4 °C, with or without agitation. One of the essential advantages of refrigerated storage of PLTs is that it reduces the risk of bacterial growth and provides a way to decrease the number of transfusion-transmitted infections . On the other hand, the main problem of cold-storage (under 15 °C) is reduced PLTs’ in vivo performance . After transfusion, they are almost immediately cleared from the bloodstream. This is initiated by changes in GPIbα, the receptor for the von Willebrand factor. The GPIbα activation induces apoptosis, initiates hemostatic responses upon rewarming, and promotes clearance of PLTs from the circulation, which is the reason why cold storage is excluded from routine procedures of transfusion medicine . The molecular mechanism of this process is already well known. In 2012, Gitz et al. examined the change in GPIbα distribution using Förster resonance energy transfer by time-gated fluorescence lifetime imaging microscopy. Their results showed that cold storage induced deglycosylation of the GPIbα ectodomain, exposing N-acetyl-D-glucosamine residues sequestered with GM1 gangliosides in lipid rafts. The study aimed not only to show the molecular mechanism of the deglycosylation of GPIbα but also to find a solution to protect PLTs from this process. It was concluded that inhibition of GPIbα clustering by the depletion of N-acetyl-2,3-dihydro-2-deoxyneuraminic acid and arachidonic acid provides a simple means to prevent damage that compromises the recovery and survival of cold-stored PLTs . Recently, Marini et al. published the results of in vitro and in vivo experiments, examining the impact of apoptosis inhibition on the hemostatic functions and survival of cold-stored PLTs. They observed that blocking the apoptotic signal transduction induced by GPIbα clustering or the activation of caspase 9 did not negatively affect the functionality of cold-stored PLT. Indeed, inhibiting the apoptotic signal mediated by GPIbα clustering with a RhoA inhibitor better preserved the release of δ granules, PLT aggregation, adhesion, and the ability to form stable clots compared to the control group. They also noted a significant improvement in the half-life of refrigerated PLTs upon inhibition of the intracellular signal induced by GPIbα clustering. The presented results provide hope that combining cold storage with apoptosis inhibition could be a promising strategy to extend the storage time of refrigerated PLTs without compromising their hemostatic functions or survival . Cold-stored PLTs undergo a process similar to agonist-induced activation; however, the similarity between these two events seems to be restricted to the final stages of the signaling cascade, making our understanding of cold activation intriguing and challenging. Cold storage can disrupt calcium ion pumps and channels in the PLT membrane, resulting in changes to intracellular calcium levels. This altered calcium homeostasis has the potential to impair PLT activation pathways, thereby affecting downstream signaling cascades essential for proper PLT function.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p24
|
PMC11277025
|
sec[2]/sec[1]/p[1]
|
3.2. Cold Storage
| 4.433594 |
biomedical
|
Review
|
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Furthermore, PLT cytoskeletal elements, particularly microtubules and microfilaments, play a crucial role in various PLT functions, such as shape change, granule release, and aggregation. The reduced temperature during cold storage may lead to disorganization of the actin cytoskeleton, which can impact both PLT structure and function. Additionally, cold storage influences PLT-PLT interactions as well as the formation of PLT aggregates. Altered membrane properties, receptor expression (inter-alia CD62P, CD63, GPIbα, GPαIIβ3), and changes in the cytoskeleton can modulate the strength and stability of PLT aggregates, which are pivotal for effective clot formation. Low temperatures also trigger the activation of PLT through processes like the release of PLT granule contents and the presentation of phosphatidylserine. Moreover, cold temperatures exert considerable influence on the metabolic processes of stored PLTs. Overall, understanding the physiological consequences of cold storage on PLTs is essential for optimizing storage conditions and preserving their functionality, ensuring their efficacy in transfusion therapies and hemostatic processes . Despite these limitations and concerns, the FDA recently issued guidance that includes a notice of exceptions and alternatives for conventional PC storage methods. This guidance permits the storage of apheresis PLTs at 1 to 6 °C for up to 14 days from the date of collection as an alternative procedure for the treatment of active bleeding in cases where conventional PLTs are unavailable, or their use is impractical . A potential application of cold-stored PLTs may involve their use during cardiac surgeries. In such cases, the prolonged survival of PLTs may not be as crucial as their functionality during the procedure and up to 24 h afterward when hemodynamic balance is disrupted .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p25
|
PMC11277025
|
sec[2]/sec[2]/p[0]
|
3.3. Cryopreservation
| 4.261719 |
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In this method, PLTs, after supernatant removal, are stored at −80 °C with the addition of dimethyl sulfoxide (DMSO). According to the procedures, cryopreserved PLTs (CPP) can be stored for up to 2 years. PLTs are rapidly transfused after thawing and resuspension in an adequate saline solution . CPPs show hemostatic functionality, and their clinical short-term efficiency has been proven , but they do not meet the FDA survival criteria for 24 h. In fact, cryopreservation induces undesirable processes in PLT, such as increased activation, unresponsiveness to agonist stimulation, and decreased levels of GPIbα, GPVI, and integrin αIIbβ . Furthermore, in 2017, Eker et al. observed strong PMP formation during cryopreservation, while there was no difference in PLT numbers before and after the procedure. Significantly higher and earlier thrombin formation occurred in the samples. At the same time, the viability of PLTs was reduced . Similar effects, such as the formation of microparticles and weakened aggregative capacity, were observed by Gavioli et al. Furthermore, it was noted that within the first 3 h after thawing, irreversible changes occur in the biomolecular structure of CPP ( Table 4 ).
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p26
|
PMC11277025
|
sec[2]/sec[2]/p[1]
|
3.3. Cryopreservation
| 3.921875 |
biomedical
|
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However, there are clinical trials that aim to implement PLT cryopreservation technology, for example, by the Australian and New Zealand Intensive Care Research Centre . In this phase III multicenter, blinded, randomized controlled clinical non-inferiority trial, it is investigated whether CPP will be at least as effective and safe as conventional liquid-stored PLTs in treating active bleeding due to surgery. The positive outcome of these studies will result in a reduction in overall PC wastage, allow smaller hospitals to provide PLT transfusions, and improve the cost-efficiency of both storage and surgical procedures.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p27
|
PMC11277025
|
sec[2]/sec[2]/p[2]
|
3.3. Cryopreservation
| 4.15625 |
biomedical
|
Review
|
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The effects of storage conditions on therapeutic PLT products at room temperature (5–7 days), cold (5–7 days), or frozen (months-years) when compared with PLTs present in fresh PRP were reviewed by Hegde et al. . Their studies indicated that, overall, room temperature storage preserves more PLT characteristics, whereas cold and DMSO frozen storage leads to various functional and structural impairments. While room temperature storage maintains normal PLT size and volume, it does reduce pH and delay clot formation and thromboxane A2 production, although swirling ability is preserved. In contrast, cold storage decreases PLT count and volume, mildly reduces pH, and increases microparticle (MP) content and granule secretion while improving aggregation response. Cryopreservation also decreases PLT count and volume, maintains a mildly reduced pH, and significantly increases MP content, causing cytoskeletal damage. Across all storage conditions, phosphatidylserine externalization, membrane CD40L, and Factor V binding increase. ATP production efficiency is poor in room temperature storage and modestly reduced in both cold and cryopreserved storage . However, current clinical research indicates that cold-stored PLTs have the worst survival and recovery rates after 2 and 7 days of storage, with significantly better outcomes for CPP and room temperature-stored PLTs, with a slight preference towards the latter . These findings emphasize the need for careful consideration of storage conditions to optimize the therapeutic potential of PLT products.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p28
|
PMC11277025
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|
3.4. Lyophilized Platelets
| 3.982422 |
biomedical
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Lyophilized platelets (LPs) have undergone lyophilization, a process involving freezing and drying under reduced pressure. This converts the cells into a dry powder, allowing for their long-term storage at room temperature with a shelf life of up to 24 months. Over the years, researchers have refined and modified the lyophilization process to develop the most effective procedures. PLTs were most commonly fixed with paraformaldehyde and then lyophilized in a solution of serum albumin . Protocols involving lyophilization with or without a cryoprotectant have been reported .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p29
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PMC11277025
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3.4. Lyophilized Platelets
| 3.955078 |
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The efficacy of LPs in PLT adhesion and bleeding reduction in thrombocytopenic models has been evaluated in numerous studies . Preliminary research has led to clinical trials assessing the safety of LPs in healthy patients . This clinical trial, which has been completed, was a randomized, blinded, dose-escalation safety trial of autologous thrombosomes, and no serious adverse events were reported. The latest ongoing trials aim to evaluate the efficacy of LPs during cardiopulmonary bypass surgery .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
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3.4. Lyophilized Platelets
| 4.007813 |
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The evaluation of LPs has also been conducted using animal models. In a study by Gogg et al., the effects of CPP and LPs were compared in a canine model . Most of the dogs had primary or secondary immune thrombocytopenia. The study indicates that LPs are comparable to CPP in managing bleeding in thrombocytopenic dogs. Within the acute period (1 h post-transfusion), LPs might be more effective than CPP in reducing bleeding scores and preventing a drop in hematocrit, although the clinical significance of these minor differences remains unclear. Future research should aim to compare LPs with fresh PC or fresh whole blood, assess LPs’ effectiveness in scenarios such as trauma and intraoperative bleeding, and explore the potential benefits of combining LPs with other hemostatic treatments, including antifibrinolytic agents .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p31
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PMC11277025
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|
3.4. Lyophilized Platelets
| 4.171875 |
biomedical
|
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To enable the use of LPs beyond thrombocytopenic patients, it was necessary to investigate the interactions between LPs and normal PLTs. In 2022, Schnoor et al. published a study characterizing LPs in comparison to unaltered PLTs . The study analyzed the activity of key PLT receptors, phosphatidylserine exposure on the PLT surface, and fibrinogen binding capacity. Additionally, the interaction between LPs and PLTs was assessed ex vivo through aggregation and adhesion experiments. These experiments shed new light on the mechanisms of LP and PLT interactions. The analysis indicated that LPs retained the receptors responsible for adhesion. A significant portion of LPs exhibited increased phosphatidylserine exposure and fibrinogen binding. The evidence clearly demonstrates that these LPs interact with untreated PLTs and significantly impact their function. Interestingly, the interactions have opposite effects on aggregation versus adhesion. LPs have a robust anti-PLT effect in terms of aggregation, leading to significant decreases in PLT aggregation when LPs are mixed with untreated PLTs. This trend was consistent across different PLT counts, indicating that LPs inhibit the aggregation function of untreated PLTs .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p32
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|
3.5. Platelet Additive Solutions (PASs)
| 3.953125 |
biomedical
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0.0010175704956054688
] |
The additive solutions are electrolyte solutions with different types of specific ingredients that could modify PLT quality. The popularity of PASs has continuously increased in recent years. The idea of using PASs was to store as small an amount of plasma with PLTs as possible. Therefore, PAS is used to replace plasma as a storage medium for PLTs. Due to the use of the procedure, some benefits are observed. First, a reduction in the amount of plasma transfused with PLTs is possible, which means a lower risk of allergic transfusion reactions with equivalent clinical efficacy to control bleeding . Another advantage is that photochemical treatment for the inactivation of bacteria and other pathogens can be applied, and as a result, it improves storage conditions. PASs are used for all types of stored PLTs (obtained by both apheresis and BC method). Nowadays, the storage medium is composed of 20–50% plasma and 50–80% PASs .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p33
|
PMC11277025
|
sec[2]/sec[4]/p[1]
|
3.5. Platelet Additive Solutions (PASs)
| 4.003906 |
biomedical
|
Study
|
[
0.99951171875,
0.0000902414321899414,
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[
0.9873046875,
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0.00012373924255371094
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There are several types of PAS. The simplest is PAS-II. It contains only sodium chloride, sodium citrate, and sodium acetate. PAS-II with phosphate as a buffer is named PAS-III. Due to phosphate in the solution, the metabolism of glucose increases, leading to higher lactate production . To reduce the adverse effects of PAS-III potassium and magnesium were added and named PAS-IIIM, which is an effective substitute for plasma . Additionally, Aurich et al. 2022 demonstrated that cold-induced cytoskeletal disorganization can be prevented by storage media supplementation with at least 10 mM magnesium ions (Mg 2+ ) .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p34
|
PMC11277025
|
sec[2]/sec[4]/p[2]
|
3.5. Platelet Additive Solutions (PASs)
| 4.019531 |
biomedical
|
Study
|
[
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[
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New solutions for PAS components are constantly sought. One of the recent discoveries is the positive effect of resveratrol on stored PLTs. The addition of this natural ingredient at a concentration of 10 μM contributed to the reduction of PLT activation and diminished free mtDNA during 7 days of storage , which is a direct marker of oxidative stress . Hence, resveratrol could serve as a potential additive to enhance the preservation quality of stored PCs.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p35
|
PMC11277025
|
sec[2]/sec[4]/p[3]
|
3.5. Platelet Additive Solutions (PASs)
| 3.035156 |
biomedical
|
Study
|
[
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[
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Multiple studies have shown a positive impact of different PAS on PLTs in vitro quality, but only a few authors investigated the parameters of PAS-treated PLTs after transfusion (in vivo) . This type of research is clinically essential and should be studied in great depth ( Table 5 ).
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p36
|
PMC11277025
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sec[2]/sec[5]/p[0]
|
3.6. Pathogen Reduction Technologies—Is It a Blessing?
| 2.552734 |
biomedical
|
Review
|
[
0.9853515625,
0.0017833709716796875,
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[
0.033721923828125,
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] |
Currently, three pathogen inactivation systems are commercially available to produce pathogen-reduced PCs. All of them use UV light in the presence or absence of a photosensitizer. These technologies were widely reviewed in the literature , which is why only key issues necessary for the context of this review are provided.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p37
|
PMC11277025
|
sec[2]/sec[5]/p[1]
|
3.6. Pathogen Reduction Technologies—Is It a Blessing?
| 4.136719 |
biomedical
|
Study
|
[
0.99951171875,
0.00015532970428466797,
0.00016629695892333984
] |
[
0.99658203125,
0.0011749267578125,
0.0022296905517578125,
0.00011420249938964844
] |
INTERCEPT™ Blood System (Cerus Corporation, Concord, CA, USA) is a PI system that combines the use of amotosalen (a psolaren) and UVA light. Amotosalen can penetrate cellular and nuclear membranes. It can develop a noncovalent link between pyrimidine bases in DNA and RNA chains. A photochemical reaction is induced by exposition to UVA light (320–400 nm) and causes the transformation of the preexisting link into an irreversible covalent bond, preventing DNA replication and RNA transcription. The results of in vitro and in vivo studies have not demonstrated any toxicologically relevant effects of PCs prepared by the INTERCEPT™ system . However, some publications reported that upon INTERCEPT™ treatment, a reduction in PLT function is observed . INTERCEPT™ has been shown to deregulate the expression of anti-apoptotic genes, modulate the PLT characteristics and aggregation response to physiological agonists, and the level of activation . Moreover, INTERCEPT™ can modulate the PLT mRNA transcriptome .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11277025_p38
|
PMC11277025
|
sec[2]/sec[5]/p[2]
|
3.6. Pathogen Reduction Technologies—Is It a Blessing?
| 3.767578 |
biomedical
|
Other
|
[
0.99853515625,
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[
0.46923828125,
0.52734375,
0.00255584716796875,
0.0006227493286132812
] |
Another PI system is MIRASOL ® (Terumo BCT, Lakewood, CO, USA). In this method, riboflavin is a photosensitizer for generating reactive oxygen species (ROS) upon UVA/UVB (270–360 nm) irradiation, which damages guanidine nucleotide bases. Riboflavin is considered safe and does not need to be removed after illumination .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p39
|
PMC11277025
|
sec[2]/sec[5]/p[3]
|
3.6. Pathogen Reduction Technologies—Is It a Blessing?
| 3.667969 |
biomedical
|
Other
|
[
0.99853515625,
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[
0.1484375,
0.84765625,
0.0031223297119140625,
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The third commercially available PI system is THERAFLEX (MacoPharma, Tourcoing, France), which uses a method based on UVC light in combination with strong agitation. Strong agitation facilitates light penetration, and the whole process does not require a photosensitizer. UVC radiation acts directly on nucleic acids by inducing pyrimidine dimers and blocking DNA replication .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p40
|
PMC11277025
|
sec[2]/sec[5]/p[4]
|
3.6. Pathogen Reduction Technologies—Is It a Blessing?
| 3.998047 |
biomedical
|
Study
|
[
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[
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Pathogen reduction technology demonstrates high effectiveness in the inactivation of most tested pathogens, including West Nile virus , Zika virus , dengue virus , and hepatitis-E virus . INTERCEPT™ demonstrates a higher inactivation capacity for the bovine viral diarrhea virus and pseudorabies virus than MIRASOL ® . Both technologies show similar log reductions for hepatitis-A virus and porcine parvovirus. Due to the applied chemistry, PI systems can inactivate pathogens that contain nucleic acids. As a consequence, they are ineffective for prions and transmission of variant Creutzfeldt–Jakob disease .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p41
|
PMC11277025
|
sec[2]/sec[5]/p[5]
|
3.6. Pathogen Reduction Technologies—Is It a Blessing?
| 3.871094 |
biomedical
|
Study
|
[
0.99951171875,
0.00016891956329345703,
0.0002982616424560547
] |
[
0.96484375,
0.003711700439453125,
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The other side of PI systems also warrants consideration, as they have not undergone sufficient testing. Moreover, the complete molecular mechanism behind these systems remains unclear. It is already established that PI treatment can result in various forms of damage to PLTs, ranging from changes in membrane integrity and signaling pathways to impaired functionality of miRNAs. This damage has the potential to diminish the recovery and survival rates in healthy patients ( Table 6 ). Currently, several clinical trials utilizing PI systems are underway, focusing on whole blood and red blood cells .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p42
|
PMC11277025
|
sec[2]/sec[5]/p[6]
|
3.6. Pathogen Reduction Technologies—Is It a Blessing?
| 3.699219 |
biomedical
|
Study
|
[
0.9990234375,
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[
0.67578125,
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] |
Based on the research conducted thus far, it is evident that PI-treated PLTs exhibit notable differences compared to untreated ones. To comprehensively understand the functional alterations in PLTs, it is imperative to conduct clinical trials involving the transfusion and further in vivo monitoring of cells treated with pathogen reduction technology. Hopes for a thorough explanation of the molecular mechanism of PI systems give proteomic analyses and metabolomics .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p43
|
PMC11277025
|
sec[2]/sec[5]/p[7]
|
3.6. Pathogen Reduction Technologies—Is It a Blessing?
| 3.910156 |
biomedical
|
Study
|
[
0.99853515625,
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[
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] |
Considering solely the inventory management aspects of blood products, PRT can contribute to waste reduction. This was demonstrated, among others, in the study by Fachini et al. 2021. The authors proved that only through the implementation of PI systems at Sírio-Libanês Hospital in São Paulo, Brazil, the discard rates of blood PCs decreased from 6% to 3%. A further decline, to 1.2%, was noted after extending the storage time to 7 days. A significant decrease in adverse transfusion events associated with the implementation of PRT was observed .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277025_p44
|
PMC11277025
|
sec[3]/sec[0]/p[0]
|
4.1. Induced Pluripotent Stem Cells
| 4.378906 |
biomedical
|
Review
|
[
0.99853515625,
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[
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COVID-19 pandemic restrictions, like lockdowns, reduced societal mobility, and sanitary and epidemiological requirements have slowed and reduced the number of donations, which has influenced a shortage of blood products. The pandemic has drawn significantly greater attention to the ex vivo production of transfusable blood cells. Beyond the pandemic, long weekends, holidays, and vacations also contribute to a decrease in blood donations. Concerns about PLT product supplies had already arisen due to their short shelf life (up to 7 days), supply–demand imbalance in aging societies of developed countries, and alloimmune-mediated PLT transfusion refractoriness. Consequently, there are high expectations for ex vivo products to address these issues. One approach involves using artificial components, such as liposomes, to create particles that mimic the function of PLTs. However, this solution has so far been ineffective in vivo, as the body still recognizes the transfused preparations as foreign, leading to increased aggregation, which is hazardous for patients. Another promising approach under investigation is the differentiation of PLTs from stem cells, particularly induced pluripotent stem cells (iPSCs) in the laboratory. Human iPSCs were first used in 2006 . Since then, interest in using these cells to produce PLTs has steadily increased due to their self-renewal capacity and pluripotency. They hold significant potential for use in transfusion medicine and other therapies. One of the key advantages of iPSC-derived PLTs is their potential for large-scale production, providing a sustainable and scalable source of PLTs for patients in need of transfusion. These cells enable the delivery of patient-specific products or adjustments to desired phenotypic characteristics.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p45
|
PMC11277025
|
sec[3]/sec[0]/p[1]
|
4.1. Induced Pluripotent Stem Cells
| 4.265625 |
biomedical
|
Study
|
[
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[
0.994140625,
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One of the major challenges in the potential clinical use of iPSCs is the establishment of an appropriate process for generating blood cells, including PLTs. This is a significant challenge because approximately 10 11 cells are transfused to a patient during a single transfusion. Without gene transduction, MKs practically do not proliferate. Therefore, the development of a cell source capable of expansion is crucial for PLT production. Feng et al., in 2014, achieved the generation of universal PLTs from iPSCs with a deletion of the β2-microglobulin gene and demonstrated that a single MK precursor generates approximately six PLTs . Moreau et al. achieved chemically defined large-scale production of MKs from iPSCs, where a single MK released approximately five PLTs . Currently, a single cell from the human adipose-derived mesenchymal stromal/stem cell line (ASCL) generates from 5 to 10 ASCL-PLTs . ASCL-PLTs have similar characteristics to peripheral PLTs and may additionally serve as mesenchymal-like cells. Tozawa et al. investigated the in vivo dynamics of ASCL-PLTs and standard PCs by administering these cells to irradiated immunosuppressed NSG mice (2.0 Gy, for 7 days). Blood samples from transfused mice were analyzed before the procedure and at 30 min, 2, 4, 6, and 24 h post-transfusion. The kinetics of ASCL-PLTs mirrored that of standard PCs. According to the study, no spontaneous PLT aggregation before agonist-induced stimulation was observed; there is a possibility that ASCL-PLTs may lead to thrombotic complications. The published protocol represents a straightforward technique, thereby bolstering the clinical implementation of the methodology. Thus, it is important to conduct further studies to optimize ASCL-PLTs as candidates for clinical applications .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11277025_p46
|
PMC11277025
|
sec[3]/sec[0]/p[2]
|
4.1. Induced Pluripotent Stem Cells
| 4.296875 |
biomedical
|
Study
|
[
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[
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Recently, the first application of iPSC-PLTs in human circulation in vivo. It was conducted in the iPLAT1 study . During the study, a clinical-grade iPSC-PLT production system was established. This study was designed as a single-center, open-label, uncontrolled dose-finding study of autologous iPSC-PLTs. The study involved only one patient, which may not reflect the impact of the produced cells on the population. The patient, a 50-year-old woman with aplastic anemia and allo-PTR immunization due to human platelet antigen (HPA)-1a antibodies, had not received transfusions previously, suggesting immunization occurred during pregnancy. Allo-PTR poses a significant clinical challenge as the PLT count does not rise even an hour after transfusion. Typically, such patients are treated with HLA class I or HPA-matched blood products. However, such products are not always available at blood centers, especially in emergencies. The same applies to autologous iPSC-PLTs produced for a specific patient. Their preparation time is 7–12 days . During the iPLAT1 study, the patient received different doses of iPSC-PLTs. No clinically significant symptoms were observed after the administration of all doses. At the end of the one-year observation period following the last dose, the external Committee for Evaluation of Efficacy and Safety concluded that autologous iPSC-PLT administration was safe for the patient. Unfortunately, even after the transfusion of the maximum final dose, there was no significant increase in the patient’s PLT count. Moreover, shortly after the transfusion, a higher concentration of the d-dimer clotting marker was observed, which may suggest unexpected coagulation of iPSC-PLTs . The research represents a notable advancement in the clinical utilization of iPSC-PLTs, facilitating the introduction of allogeneic alternatives and, leveraging their prospective advantages, augmenting the breadth of blood transfusion medicine.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p47
|
PMC11277025
|
sec[3]/sec[1]/p[0]
|
4.2. Photobiomodulation
| 4.15625 |
biomedical
|
Study
|
[
0.99951171875,
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[
0.99072265625,
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Studies have demonstrated that photobiomodulation using near-infrared (NIR) light, also known as low-level light therapy (LLLT), can significantly impact the quality and functionality of PLTs during storage and various medical procedures. Recently, Bontekoe et al. showed that the NIR treatment of PCs in PAS-E resulted in reduced PLT activation, evidenced by lower CD62P expression and reduced Annexin A5 binding. Additionally, there was decreased lactate production and higher pH levels, indicating a modulation of glycolysis, potentially through mitochondrial aerobic metabolism . This aligns with the findings of Yang et al. and Zhang et al. , who studied the effects of LLLT on immune thrombocytopenia in mice. They found that LLLT could alleviate immune thrombocytopenia by enhancing mitochondrial biogenesis in MKs and preserving mitochondrial function in PLTs, leading to increased PLT production and functionality .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p48
|
PMC11277025
|
sec[3]/sec[1]/p[1]
|
4.2. Photobiomodulation
| 4.046875 |
biomedical
|
Study
|
[
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[
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Moreover, in vitro studies on whole blood performed by Walski et al. demonstrated that NIR PBM could reversibly inhibit PLT activation in a dose-dependent manner and reduce hemolysis, suggesting protective effects on both red blood cells and PLTs , which could be applied not only in storage but also in extracorporeal circulation systems .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p49
|
PMC11277025
|
sec[3]/sec[1]/p[2]
|
4.2. Photobiomodulation
| 3.880859 |
biomedical
|
Study
|
[
0.99951171875,
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[
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In summary, NIR PBM exhibits significant potential in improving the quality and stability of stored PLTs through modulation of mitochondrial metabolism and protection against oxidative stress. These findings suggest promising applications for NIR and LLLT in enhancing PLT storage conditions and extending PLT shelf life in transfusion medicine.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p50
|
PMC11277025
|
sec[3]/sec[2]/p[0]
|
4.3. Proteomic Analysis (Proteomics)—A Potential Biomarker for Platelets
| 3.783203 |
biomedical
|
Other
|
[
0.99853515625,
0.0009899139404296875,
0.0006394386291503906
] |
[
0.044189453125,
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Proteomic analysis refers to the systematic identification and quantification of the complete set of proteins (the proteome) of a biological system (cell, tissue, organ, biological fluid, or organism) at a specific point in time . For proteomic analysis, the most often used technique is mass spectrometry .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277025_p51
|
PMC11277025
|
sec[3]/sec[2]/p[1]
|
4.3. Proteomic Analysis (Proteomics)—A Potential Biomarker for Platelets
| 3.068359 |
biomedical
|
Other
|
[
0.998046875,
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[
0.377685546875,
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0.0009150505065917969
] |
Despite PLTs being unnucleated cells, they contain mRNA and possess all the necessary tools required for protein translation, inherited from their precursor cells, MKs. This characteristic has recently made PLTs the focus of proteomic studies .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p52
|
PMC11277025
|
sec[3]/sec[2]/p[2]
|
4.3. Proteomic Analysis (Proteomics)—A Potential Biomarker for Platelets
| 4.078125 |
biomedical
|
Study
|
[
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[
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In 2017, Rijkers et al. studied changes in PLTs during prolonged storage in standard conditions using label-free quantitative mass spectrometry and identified 21 proteins that showed altered expression levels. Although the functionality of examined PLTs decreased, the changes in protein levels were relatively minor, suggesting that overall protein composition was only moderately affected. Additionally, the response to agonists was found to be reduced. During the study, no specific markers distinguishing ‘young’ or ‘old’ PLTs were identified. Only certain assumptions were made. For instance, it was observed that the expression of protein S100A9 rapidly declined on day 5 of storage, which may be indicative of young PLTs. Moreover, initial findings suggest that A2M, IGM, and GYG1 could potentially serve as biomarkers for ‘aged’ PLTs, but further investigation is required to validate this hypothesis .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p53
|
PMC11277025
|
sec[3]/sec[2]/p[3]
|
4.3. Proteomic Analysis (Proteomics)—A Potential Biomarker for Platelets
| 4.054688 |
biomedical
|
Study
|
[
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[
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Heililahong et al. conducted a research study where they performed a whole transcriptome analysis on stored PLTs on day 0, day 2, and day 4. They characterized mRNA, lncRNA, and circRNA profiles and discovered that differentially expressed RNA is closely related to PLT function, metabolism, DNA repair, cell cycle, and apoptosis. They identified certain candidate mRNAs as potential biomarkers for storage damage and observed that most of the pathways were related to DNA repair .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p54
|
PMC11277025
|
sec[3]/sec[2]/p[4]
|
4.3. Proteomic Analysis (Proteomics)—A Potential Biomarker for Platelets
| 4.109375 |
biomedical
|
Review
|
[
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[
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To analyze proteomic changes, various methods are employed. Examples include differential gel electrophoresis (DIGE), isotope-coded affinity tagging (ICAT), and isotope tagging for relative and absolute quantification (iTRAQ) . In 2008, it was shown that utilizing different proteomic analysis techniques coupled with mass spectrometry yields varying results . Only 5% of the findings were consistent across all methods used. Consequently, it is crucial to exercise caution when interpreting results obtained from a single-method analysis; nevertheless, proteomics, as an emerging and fascinating tool, holds promise for enhancing our understanding of the formation of PSL. It is also evident that the use of omics technologies can better match individual PCs to patients, potentially improving the efficiency and safety of transfusions. Through omics, also including metabolomics and transcriptomics, it is possible to gain a deeper understanding of the factors affecting the quality of stored PLTs and subsequently develop strategies to enhance their stability and extend their shelf life . This may include modifications in storage procedures, the addition of appropriate protective substances, or genetic modifications.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p55
|
PMC11277025
|
sec[3]/sec[3]/p[0]
|
4.4. miRNA Changes and PLTs Storage Conditions
| 3.416016 |
biomedical
|
Other
|
[
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[
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0.035858154296875,
0.0012903213500976562
] |
miRNA is a small regulatory, single-stranded RNA (19–24 nucleotides) and was discovered in 1993. From that time on, more than 2500 have been defined . They play a significant role in the post-transcriptional regulation of protein expression, affecting approximately one-third to half of human genes .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p56
|
PMC11277025
|
sec[3]/sec[3]/p[1]
|
4.4. miRNA Changes and PLTs Storage Conditions
| 4.171875 |
biomedical
|
Study
|
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[
0.9970703125,
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] |
Recently, it was shown that miRNA plays an essential role in PLT regulatory functions in synthesizing proteins responsible for PLT activation and thrombus formation . In 2009, Landry et al. found that P2Y 12 mRNA is associated with Ago2·miRNA complexes in PLTs. They concluded that miRNAs might potentially exert coordinated and/or synergistic effects to regulate P2Y 12 mRNA translation through its 3′untranslated regions (3′UTR). In such cases, miRNAs could play a crucial role in modulating PLT function .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p57
|
PMC11277025
|
sec[3]/sec[3]/p[2]
|
4.4. miRNA Changes and PLTs Storage Conditions
| 4.082031 |
biomedical
|
Study
|
[
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[
0.9970703125,
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] |
A comprehensive study was conducted by Osman et al. in 2014 to investigate the effects of three PI systems, namely INTERCEPT™, MIRASOL ® , and gamma-irradiation, on the levels of miRNAs and mRNAs in PLTs stored in blood banks as well as their impact on PLT activation and function. The results revealed that 6 of the 11 studied miRNAs were already reduced by INTERCEPT™ on day 1. Both MIRASOL ® and INTERCEPT™ treatments resulted in a decrease in PLT count on day 1 of storage. Additionally, only the INTERCEPT™-treated PLTs exhibited a decrease in anti-apoptotic mRNAs within the first 24 h. The authors suggested that there is probably some specific mechanism related to miRNA changes in PI-treated PLTs. miRNA technology is relatively new compared to earlier designed PI systems, so it is advisable to stop the implementation of PI until its safety for miRNA is established.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p58
|
PMC11277025
|
sec[3]/sec[3]/p[3]
|
4.4. miRNA Changes and PLTs Storage Conditions
| 4.0625 |
biomedical
|
Study
|
[
0.99951171875,
0.00018334388732910156,
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] |
[
0.9990234375,
0.0002720355987548828,
0.0005011558532714844,
0.0000655055046081543
] |
Further investigations demonstrated that PLTs and PMPs possess distinct miRNA profiles. The authors suggest a specific miRNA loading into MPs during PLT activation . Exposure of PLTs to INTERCEPT™ caused alterations in miRNA levels within PMPs, while MIRASOL ® treatment showed no significant effect on the miRNA profile in PLTs . The authors suggested that changes in certain miRNA levels within PMPs derived from INTERCEPT™-exposed PLTs might contribute to the observed increased bleeding in some recipients.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p59
|
PMC11277025
|
sec[3]/sec[3]/p[4]
|
4.4. miRNA Changes and PLTs Storage Conditions
| 3.820313 |
biomedical
|
Study
|
[
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[
0.99755859375,
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] |
Another group of researchers discovered that pathogen reduction technology may influence the miRNA profile of PMPs and induce overrepresentation of specific miRNAs . They compared MIRASOL ® and INTERCEPT™ treatments, and the changes in miRNA were found in both cases. The conclusion from this study was to be careful with transfusing modified PLTs to patients, as the long-term consequences of using such altered materials remain unknown.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p60
|
PMC11277025
|
sec[4]/p[0]
|
5. Big Data and Statistical Analysis in PLT Storage Management
| 3.541016 |
biomedical
|
Other
|
[
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] |
[
0.064453125,
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Minimizing the wastage of blood and blood products is a crucial requirement all over the world. Two goals for a blood management system are the most important: ensuring product availability even in emergencies and reducing wastage resulting from product expiration. Managing PLT production is particularly challenging due to its short shelf life, leading to high rates of wastage and significant economic costs. Moreover, overproduction is common due to the inability of patients requiring PLT transfusion to wait for PC production.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p61
|
PMC11277025
|
sec[4]/p[1]
|
5. Big Data and Statistical Analysis in PLT Storage Management
| 4.132813 |
biomedical
|
Study
|
[
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] |
[
0.99853515625,
0.0003178119659423828,
0.0011348724365234375,
0.00012230873107910156
] |
The study conducted in 2016 by Sekhar et al. provides compelling evidence that effective management control can lead to improved concentrations of PC . The authors presented data from a patient blood management program that involved the implementation of a service improvement initiative through the introduction of a ‘platelet coordinator’ role. The primary objective was to optimize the utilization of PLTs in a large, complex tertiary care hospital within the National Health Service setting. The study spanned three years and demonstrated a significant reduction in the supply and costs of PLTs. Specifically, between 2012/2013 and 2014/2015, there was a 21% decrease in both the number of PLT units supplied to the institution and the associated expenditure on PLTs. This reduction in PLT supply and costs was attributed to the implementation of enhanced strategies in waste management and stock control. These measures led to a more efficient utilization of the provided PLTs within the hospital. Despite an increase in overall hospital activity during the study period, the hospital managed to maintain stable PLT issue figures. Notably, the improvement in wastage control exceeded the improvements observed in the expenditure on PLTs, suggesting the successful optimization of PC usage .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277025_p62
|
PMC11277025
|
sec[4]/p[2]
|
5. Big Data and Statistical Analysis in PLT Storage Management
| 1.892578 |
biomedical
|
Other
|
[
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[
0.00890350341796875,
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] |
Each country has its unique blood management system, which can vary in complexity depending on specific circumstances. Blood donation centers may support multiple hospitals or a dozen or more, making direct replication of solutions across countries unfeasible. In each case, it is necessary to conduct research and adapt the model to individual needs.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277025_p63
|
PMC11277025
|
sec[4]/p[3]
|
5. Big Data and Statistical Analysis in PLT Storage Management
| 2.384766 |
biomedical
|
Other
|
[
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[
0.03082275390625,
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Nowadays, leveraging data analysis for optimizing PC production processes has become indispensable for the efficient utilization of healthcare resources. While the concept of modeling and estimating the demand for PCs is not new, with the first published studies dating back to the 1980s and 1990s , we now have access to greater computing power and artificial intelligence. Moreover, the ability to transfer and archive large amounts of data has become crucial for any modeling or machine learning endeavors.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p64
|
PMC11277025
|
sec[4]/p[4]
|
5. Big Data and Statistical Analysis in PLT Storage Management
| 4.019531 |
biomedical
|
Study
|
[
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[
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In 2016, Pérez et al. published an article introducing the mathematical model they developed to enhance the management of PC production, aiming to make decision-making more rational and less empirical . The data used for the model were derived from historical records of PCs produced, transfused, and discarded in the Basque Country in 2012. The model assumes a normal distribution of demand on each day of the week throughout the year, as previously observed in 2012. The authors presented an Excel spreadsheet where the estimation of the daily production of PCs was possible. They validated the model using real production data in 2013. The conclusion drawn was that the model served as a useful support tool during PC production, albeit with insufficient precision. Nevertheless, the results indicated a beneficial effect of modeling the data. Firstly, decisions could be made in a more rational manner rather than relying solely on empirical methods. Secondly, the model offered potential cost reductions of approximately ~0.5 million EUR annually. Lastly, the advantage of the model was that transfused units could be one day younger. The authors noted that for improved modeling precision, the implementation of a highly developed IT system is necessary. It would provide accurate information in real-time, thus enhancing the overall effectiveness of the model.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11277025_p65
|
PMC11277025
|
sec[4]/p[5]
|
5. Big Data and Statistical Analysis in PLT Storage Management
| 4.070313 |
biomedical
|
Study
|
[
0.9990234375,
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[
0.99755859375,
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Guan et al. built and validated a statistical model for blood banking that enabled the prediction of PLT usage three days in advance. The data used for the study were obtained from the Stanford Blood Centre, which supplies all blood products for two associated hospitals: Stanford Health Care and Lucille Packard Children’s Hospital. The availability of data was facilitated by the implementation of a modern hospital electronic medical records (EMR) system. The research was based on finding the link between hospital-wide patient data and clinical transfusion decisions. The developed model considered data from 29 consecutive months and successfully reduced the expiration rate from 10.5 to 3.2%. It prohibits PLT shortages by reserving a minimum of 10 PC units on the shelf each day. The authors suggest that annually in this institution, the wastage of PCs could be reduced by 950 units without compromising patient care. If the same results were transferred nationally in the United States, the healthcare system could potentially save approximately 80 million USD .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277025_p66
|
PMC11277025
|
sec[4]/p[6]
|
5. Big Data and Statistical Analysis in PLT Storage Management
| 2.490234 |
biomedical
|
Other
|
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[
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There is still limited research available in the field of big data and statistical analysis for PLT storage management with a solution ready to use. Conducting this type of study requires a substantial amount of data, and fortunately, it is becoming increasingly feasible to gather such data today. This area of research holds great promise and importance as it has the potential to significantly reduce blood and blood component wastage without relying on complex biological or additional techniques .
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
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|
sec[5]/p[0]
|
6. Conclusions
| 4.066406 |
biomedical
|
Review
|
[
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[
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In recent years, much has been performed to better understand the effects of PLT storage. This brings with it improved transfusion safety and the possibility of better use of this valuable material. During the research, the database of clinical trials was checked. As of April 2024, the database contained records of 10 active clinical trials focused on PLT storage. These studies primarily revolve around two main objectives: cold storage (six trials) and cryopreservation (three trials). Additionally, one of the clinical trials focuses on investigating the impact of PLT storage on complement activation. Some specific topics of investigation include examining the impact of CD47 expression levels on PLTs stored at different temperatures , as well as studying CD62-P expression during storage and assessing the influence of preparation techniques . Today, the world of science has access to tools and technologies that were not available a decade or so ago, offering new possibilities for the improvement of the PLT storage process. It would be worthwhile to determine whether these new findings can enhance the functionality of PLTs post-transfusion. The main goals for researchers in the next few years should be a better understanding of PSL, upgrading the banking methods of PLTs with the investigation of their in vivo quality, and employing big data modeling and artificial intelligence to estimate and predict PC production.
|
[
"Natalia Trochanowska-Pauk",
"Tomasz Walski",
"Raghvendra Bohara",
"Julia Mikolas",
"Krystian Kubica"
] |
https://doi.org/10.3390/ijms25147779
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p0
|
PMC11277037
|
sec[0]/p[0]
|
1. Introduction
| 4.113281 |
biomedical
|
Study
|
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Galloway–Mowat syndrome (GAMOS) is an extremely rare autosomal or X-linked recessive disorder that primarily affects the kidneys and central nervous system . The incidence of GAMOS is less than one in a million . At present, eleven causative genes have been identified: five KEOPS (Kinase, Endopeptidase, and Other Proteins of small Size) complex related-genes ( OSGEP , TP53RK , TPRKB , LAGE3 , and GON7 ) and six other genes ( WDR73 , WDR4 , NUP133 , NUP107 , YRDC , and PRDM15 ) .
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277037_p1
|
PMC11277037
|
sec[0]/p[1]
|
1. Introduction
| 4.160156 |
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|
Study
|
[
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OSGEP (O-sialoglycoprotein endopeptidase) was first reported to be associated with GAMOS in 2017 . To date, OSGEP variants in 45 patients from 38 families have been reported and are the most prevalent certain stemness markers of the genetic etiology of GAMOS . Most affected individuals die in early childhood, and the primary mutation type of OSGEP is a missense mutation. In a previous study, the knockout of OSGEP in zebrafish and mouse models resulted in early lethality . Such results suggest that the complete absence of OSGEP leads to cell death and that OSGEP is essential for normal growth and development in a variety of model organisms. However, research on its impact on humans is limited. Further investigation into the role of OSGEP in human cells is conducive to advancing our understanding of GAMOS.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277037_p2
|
PMC11277037
|
sec[0]/p[2]
|
1. Introduction
| 4.703125 |
biomedical
|
Study
|
[
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[
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OSGEP as a catalytic unit of the KEOPS complex participates in the biosynthesis of N6-threonylcarbamoyl adenosine (t6A), a universal transfer RNA (tRNA) modification at position 37 that decodes ANN codons in eukaryotes and archaea . tRNA modification is crucial for protein synthesis and is extensively characterized in Saccharomyces cerevisiae . The authors of several studies have linked tRNA hypomodification to elevated missense error rates and disrupted codon decoding efficiency, resulting in protein aggregation and harmful effects on the cell . A number of studies have demonstrated the role of proteostasis in mouse neural stem cells (NSCs) and their ability to exit quiescence, self-renew, and differentiate . To remove abnormal protein aggregates and maintain protein homeostasis, autophagy is employed. Autophagy plays a crucial role in maintaining neuronal homeostasis and synaptic function. In numerous neurodegenerative diseases marked by the accumulation of protein aggregates—such as Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, and amyotrophic lateral sclerosis (ALS)—this process is often dysregulated . ELP3 (Elongator Acetyltransferase Complex Subunit 2) is a susceptibility gene for ALS and affects the lifespan of some ALS patients . ELP3 is involved in the modification of the wobble uridine (U34) of tRNA, and its mutants can lead to inappropriate autophagy . The above studies suggest that tRNA modifications are closely related to protein homeostasis and autophagy. However, no study has elucidated the impact of the KEOPS complex on proteostasis and autophagy, especially in human cells.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p3
|
PMC11277037
|
sec[0]/p[3]
|
1. Introduction
| 4.101563 |
biomedical
|
Study
|
[
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[
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In the present study, we developed an OSGEP -knockdown hESC model by using lentivirus shRNA infection. Based on the hECS model, we detected the proliferation, apoptosis, and translation of stem cell markers to investigate the impact of OSGEP on hESC fate. Moreover, we examined protein synthesis rates, protein aggregation, and alterations in autophagy levels to explore the global protein changes during OSGEP deficiency.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277037_p4
|
PMC11277037
|
sec[1]/sec[0]/p[0]
|
2.1. OSGEP Knockdown Results in Reduced Proliferation and Increased Apoptosis
| 4.320313 |
biomedical
|
Study
|
[
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[
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To investigate the functional role of OSGEP in stem cells, we constructed OSGEP -knockdown (sh OSGEP ) hESCs, which demonstrated reductions of 40% and 63% in OSGEP mRNA and protein levels, respectively . Our morphological analysis results revealed that OSGEP silencing maintained the typical dome-shaped colonies but with a reduced surface area and increased intercellular spacing . The sh OSGEP hESCs showed a significantly reduced EdU staining area, indicating decreased cell proliferation . Furthermore, OSGEP knockdown led to an enlargement of the nuclei compared with the control . Our cell cycle analysis results indicated a significant increase in the proportion of cells in the G2/M phase and a decrease in the proportion of cells in the G0/G1 phase in sh OSGEP hESCs , suggesting OSGEP knockdown-induced cell cycle arrest in the G2/M phase. Additionally, our apoptosis analysis results revealed a higher cell apoptosis index in sh OSGEP hESCs . The above findings collectively highlight the relevance of OSGEP in hESC proliferation and apoptosis.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277037_p5
|
PMC11277037
|
sec[1]/sec[1]/p[0]
|
2.2. OSGEP Knockdown Affects the Expression of Stemness Markers
| 4.308594 |
biomedical
|
Study
|
[
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We performed alkaline phosphatase (AP) staining assays, in which high AP levels serve as a marker for undifferentiated states. Remarkably, we observed a significant decrease in the AP-positive population in sh OSGEP hESCs . Additionally, we investigated several genes essential for maintaining the pluripotency of hESCs, including NANOG , OCT4 , TBX3 , and ESRRB . Although NANOG and OCT4 showed no significant changes in protein and mRNA levels, a significant reduction in TBX3 expression was noted . Subsequently, we explored how the reduction in OSGEP affects the ability of hESCs to differentiate into various cell lineages. The increased expression of TBX6 in sh OSGEP hESCs suggests that the cells might differentiate toward a mesodermal lineage. Elevated levels of PECAM1 suggest that the sh OSGEP hESCs might differentiate into endothelial cells. The reduction in both OTX2 and SOX1 suggests a decrease in neural differentiation potential . The above results indicate that OSGEP knockdown affects the expression of pluripotency markers of hESCs, making them more prone to differentiation and causing a shift away from the neural lineage.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277037_p6
|
PMC11277037
|
sec[1]/sec[2]/p[0]
|
2.3. OSGEP Deficiency Perturbs Proteostasis
| 4.21875 |
biomedical
|
Study
|
[
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Given the recognized influence of tRNA modifications on proteostasis, we examined the impact of OSGEP on global protein synthesis in hESCs. The results of a puromycin incorporation assay showed that de novo protein synthesis rates were remarkably elevated in sh OSGEP hESCs . These findings were further validated in the HEK293T cell line . In sh OSGEP hESCs, numerous red punctate signals were observed in the cytoplasm , and the fluorescence intensity was significantly higher than in the control cells . We used MG-132 as a positive control, a relatively non-specific proteasome inhibitor known to accelerate the formation of perinuclear aggresomes and inclusion bodies. The MG-132 treatment group exhibited more numerous and stronger red fluorescent signals and fewer cells, indicating that the accumulation of proteins leads to cell death . The above results indicate that OSGEP depletion leads to elevated protein synthesis and the accumulation of aggregated proteins, highlighting the potential role of OSGEP in maintaining protein homeostasis in hESCs.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p7
|
PMC11277037
|
sec[1]/sec[3]/p[0]
|
2.4. OSGEP Deficiency Induces Inappropriate Autophagy
| 4.203125 |
biomedical
|
Study
|
[
0.99951171875,
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[
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To investigate the involvement of autophagy in aggregated proteins, we examined alterations in autophagy-associated protein levels. P62, a cargo protein specific to autophagosomes, forms aggregates that can be selectively degraded through autophagy . The expression levels of P62 decreased in sh OSGEP hESCs . The conversion of LC3-I to LC3-II serves as a key marker of autophagosome formation. Upon the silencing of OSGEP , we observed a notable decrease in LC3-I expression levels and a significant elevation in the LC3-II/LC3-I ratio . The above findings collectively indicate that OSGEP depletion triggers autophagy as a cellular response to counteract the accumulation of aggregated proteins.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p8
|
PMC11277037
|
sec[2]/p[0]
|
3. Discussion
| 4.046875 |
biomedical
|
Study
|
[
0.99951171875,
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[
0.99951171875,
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0.00022470951080322266,
0.00005829334259033203
] |
In the study presented herein, we observed that OSGEP knockdown in hESCs led to decreased cell proliferation and increased apoptosis. These findings are consistent with observations in OSGEP -knockdown human podocyte cell lines , indicating that impaired cell proliferation and increased apoptosis are common pathogenic features of GAMOS.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p9
|
PMC11277037
|
sec[2]/p[1]
|
3. Discussion
| 4.238281 |
biomedical
|
Study
|
[
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Cell cycle regulation is important for the self-renewal and pluripotency of hESCs . Our findings suggest that OSGEP silencing results in cell cycle arrest in hESCs, which may impact their self-renewal capacity. This finding is consistent with previous studies demonstrating that strongly mitotic tissues are highly sensitive to the loss of kae1 (the Drosophila ortholog of OSGEP ), whereas non-proliferating tissues are less affected . Additionally, the results of a previous study showed that the KEOPS complex may inhibit Mis17-Mis6 complex-mediated centromere formation in Schizosaccharomyces pombe , which could be associated with G2/M phase arrest in hESCs.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277037_p10
|
PMC11277037
|
sec[2]/p[2]
|
3. Discussion
| 4.300781 |
biomedical
|
Study
|
[
0.99951171875,
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[
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] |
The results of our puromycin incorporation assay demonstrate that OSGEP knockdown leads to increased protein synthesis in both hESCs and HEK293T cells. The observed increase in protein synthesis may be related to an increase in the frequency of leaky scanning through start codons, leading to enhanced protein translation. This phenomenon has also been observed in yeast lacking the function of SUA5/YDRC, which is involved in t6A modifications . Additionally, in yeast with kae1 mutations, the upregulation of genes related to amino acid synthesis can be observed . In our study, an abnormal increase in protein aggregates in OSGEP -knockdown hESCs was identified. This observation supports the result of increased protein synthesis, potentially due to erroneous translation products caused by defective t6A modifications. The results of previous studies have shown that abnormalities in t6A modifications can lead to increased +1 frameshift frequencies and readthrough of stop codons . Whether the increase in abnormal protein synthesis observed in OSGEP -knockdown human cells is due to the same underlying mechanism requires further investigation.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p11
|
PMC11277037
|
sec[2]/p[3]
|
3. Discussion
| 4.167969 |
biomedical
|
Study
|
[
0.99951171875,
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] |
[
0.99951171875,
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] |
In our study, we observed that OSGEP knockdown in hESCs leads to increased autophagic activity. This enhanced autophagy likely represents a cellular attempt to clear the increased protein aggregates resulting from OSGEP deficiency. Maintaining proper proteostasis is a fundamental requirement for cellular functionality and viability. Our results indicate that OSGEP knockdown affects protein synthesis, which in turn impacts the proliferation and apoptosis of hESCs, highlighting the importance of t6A modification in maintaining protein homeostasis.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11277037_p12
|
PMC11277037
|
sec[2]/p[4]
|
3. Discussion
| 4.167969 |
biomedical
|
Study
|
[
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[
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In conclusion, the results of our study elucidate the impact of OSGEP on stem cell development, demonstrating that OSGEP is crucial for maintaining the fate of hESCs. Moreover, our study results reveal that OSGEP deficiency leads to proteostasis dysregulation and autophagy activation. The above findings thus provide new insights into the pathogenic mechanisms of GAMOS syndrome. Further research is warranted to unravel the intricate mechanisms underlying the interplay between OSGEP , proteostasis, and autophagy in the context of neurodevelopmental disorders.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277037_p13
|
PMC11277037
|
sec[3]/sec[0]/p[0]
|
4.1. Cell Culture
| 4.109375 |
biomedical
|
Study
|
[
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[
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] |
H9 hESCs were regularly cultured in mTeSR Plus medium (STEMCELL Technologies, Vancouver, BC, Canada) on plates coated with Matrigel. HEK293T cells were grown in DMEM supplemented with 10% fetal bovine serum (FBS). Cell passaging was performed at 70–80% confluence. All cell cultures were maintained in a humidified incubator set at 37 °C with 5% CO 2 .
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p14
|
PMC11277037
|
sec[3]/sec[1]/p[0]
|
4.2. Lentivirus Production and Transduction of hESCs
| 4.136719 |
biomedical
|
Study
|
[
0.99951171875,
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[
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] |
To generate OSGEP -knockdown hESCs, hESCs were infected with pLKO.1 lentiviruses containing OSGEP shRNA1 (sh1), OSGEP shRNA2 (sh2), and a non-target shRNA (negative control, NC). The sequences of shRNAs used are listed in Table S1 . Briefly, HEK293T cells were transfected with the specified lentiviral vector utilizing Lipofectamine 3000 reagent (Invitrogen, Waltham, MA, USA) following the manufacturer’s guidelines. The target and packaging plasmids psPAX2 and pMD2.G (VSV-G envelope protein) were used in a 3:2:1 ratio. After 48 h, the culture medium was collected and filtered with a 0.45 μm filter (Millipore, Burlington, MA, USA). Subsequently, hESCs were incubated with the obtained lentivirus-containing medium at 37 °C for 12 h. Following an additional 24 h, cells were selected by using 1 μg/mL puromycin for 1 week.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p15
|
PMC11277037
|
sec[3]/sec[2]/p[0]
|
4.3. HESC Self-Renewal and Proliferation Analysis
| 4.136719 |
biomedical
|
Study
|
[
0.99951171875,
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[
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] |
For the colony formation assay, 1000 cells were plated in 6-well plates. Colony size was assessed on day 7. Alkaline phosphatase (AP) staining was carried out using the Alkaline Phosphatase Staining Kit for Stem Cell (Maokang Biotechnology Co., Ltd., Shanghai, China). Briefly, 2–3 days after seeding, cells were washed with PBS, fixed for 2–5 min, and then incubated with AP solution for 5–10 min at 25 °C, shielded from light. Stained colonies were examined and documented using a microscope. At least 100 colonies per sample were counted for data analysis.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277037_p16
|
PMC11277037
|
sec[3]/sec[3]/p[0]
|
4.4. Flow Cytometry Analysis
| 4.15625 |
biomedical
|
Study
|
[
0.99951171875,
0.0002161264419555664,
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] |
[
0.99755859375,
0.0018701553344726562,
0.000583648681640625,
0.00010526180267333984
] |
For cell cycle analysis, cells were collected using Accutase (STEMCELL Technologies) and fixed overnight in 70% ice-cold ethanol. After fixation, cells were stained for 30 min with a solution containing 100 μg/mL RNase A and 50 μg/mL propidium iodide (PI) (Coolaber, Beijing, China). Flow cytometry was performed with a DxP Athena™ flow cytometer, and the data were analyzed using ModFit software version 4.1. To measure apoptotic cells, the Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme, Nanjing, China) was used according to the manufacturer’s instructions. A flow cytometer and FlowJo version 10.8.1 were used for the analysis of the flow cytometric results. For the assessment of protein aggregation, cells were stained by using the Proteostat Protein Aggregation Assay Kit (Enzo Life Science, Farmingdale, NY, USA) following the manufacturer’s guidelines.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11277037_p17
|
PMC11277037
|
sec[3]/sec[4]/p[0]
|
4.5. Immunofluorescence Staining
| 4.03125 |
biomedical
|
Study
|
[
0.9990234375,
0.0005354881286621094,
0.00040435791015625
] |
[
0.81982421875,
0.1778564453125,
0.001728057861328125,
0.0007343292236328125
] |
Cells were seeded onto Matrigel-covered glass coverslips in mTeSR Plus medium. The cells were fixed in 4% paraformaldehyde for 20 min and incubated at 4 °C in a blocking buffer (PBS containing 5% BSA). The cells were stained in accordance with the manufacturer’s procedure and then washed three times in PBS.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11277037_p18
|
PMC11277037
|
sec[3]/sec[5]/p[0]
|
4.6. Real-Time Quantitative PCR
| 4.078125 |
biomedical
|
Study
|
[
0.99951171875,
0.00015115737915039062,
0.00016760826110839844
] |
[
0.998046875,
0.00135040283203125,
0.0003643035888671875,
0.00008171796798706055
] |
Total RNA was extracted using TRIzol reagent (Invitrogen). For cDNA synthesis, 2 μg of RNA was utilized with the RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific, Waltham, MA, USA). Gene expression analysis was performed using the PowerUp SYBR Green Master Mix (Applied Biosystems, Waltham, MA, USA) on an ASA-9600 RT-qPCR System (BAIYUAN GENE-TECH, Lanzhou, China). Relative gene expression was determined using the comparative cycle threshold (Ct) method and analyzed using the 2 −ΔΔCt method. The RT-qPCR primer details are provided in Table S2 .
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277037_p19
|
PMC11277037
|
sec[3]/sec[6]/p[0]
|
4.7. Measurement of Protein Synthesis by Puromycin Incorporation
| 3.876953 |
biomedical
|
Study
|
[
0.99951171875,
0.0002453327178955078,
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] |
[
0.9765625,
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0.0002582073211669922
] |
The cells were incubated for 30 min in complete culture media containing 10 μg/mL puromycin. After treatment, the cells were washed twice with PBS, collected, and lysed using loading buffer. We used an anti-puromycin antibody to detect the puromycin by Western blot.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11277037_p20
|
PMC11277037
|
sec[3]/sec[7]/p[0]
|
4.8. Western Blot Assay
| 4.066406 |
biomedical
|
Study
|
[
0.99951171875,
0.0001672506332397461,
0.0002111196517944336
] |
[
0.994140625,
0.005123138427734375,
0.0005497932434082031,
0.00012755393981933594
] |
Cell pellets were collected and lysed with a loading buffer. The lysates were denatured by heating at 100 °C for 10 min. Proteins from the whole-cell lysates were separated using SDS-PAGE and transferred to PVDF membranes (Millipore). The membranes were then incubated with specific primary antibodies, as detailed in Table S3 , and the antibody–protein complexes were visualized using secondary antibodies conjugated to horseradish peroxidase (Sigma, Tokyo, Japan) and the SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Thermo Scientific). The images were captured by the Molecular Imager ChemiDoc XRS+ System.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11277037_p21
|
PMC11277037
|
sec[3]/sec[8]/p[0]
|
4.9. Quantification and Statistical Analysis
| 3.455078 |
biomedical
|
Study
|
[
0.99951171875,
0.00016057491302490234,
0.00047278404235839844
] |
[
0.98583984375,
0.0128631591796875,
0.0010404586791992188,
0.0001990795135498047
] |
We used ImageJ 1.53a to quantify the fluorescence intensity values and Western blot (WB) grayscale values, and we used Origin 2022 for data visualization. All results are representative of at least three independent experiments. The data are presented as the mean + SEM (Standard Error of the Mean). We used a t -test to perform the significance analysis. The p -value levels are denoted as follows: n.s ≥ 0.05; * < 0.05; ** < 0.01; *** < 0.001; **** < 0.0001.
|
[
"Hua Teng",
"Siyi Chen",
"Fang Liu",
"Yanling Teng",
"Yunlong Li",
"Desheng Liang",
"Lingqian Wu",
"Zhuo Li"
] |
https://doi.org/10.3390/ijms25147889
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
39057269_p0
|
39057269
|
sec[0]/p[0]
|
1. Introduction
| 4.023438 |
biomedical
|
Study
|
[
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[
0.82373046875,
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] |
In medico-legal entomology, the duration of insect colonization on human or animal remains at a crime scene provides critical insights for accurately estimating the postmortem interval (PMI). When conditions facilitate insect access to a carcass, insects typically begin colonization during the initial stages of decomposition. This process allows for the precise estimation of the Time of Colonization (TOC) or the minimum Postmortem Interval (mPMI) . Medico-legal entomologists estimate the mPMI by analyzing larval growth duration as an indicator of the time since death or by examining the succession patterns of insects during different stages of decomposition . Additionally, they strive to understand the decomposition process through changes in the weight and physical characteristics of the remains .
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
39057269_p1
|
39057269
|
sec[0]/p[1]
|
1. Introduction
| 4.007813 |
biomedical
|
Study
|
[
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[
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The blowfly (Diptera: Calliphoridae) plays a significant role as a primary decomposer of carcasses post-mortem, contributing to the ecological processes of decay . Research into the community composition and succession of blowflies can provide valuable insights into the timing and environmental conditions of colonization events, enhancing the accuracy of forensic analyses . Blowfly species diversity and distribution are influenced by various environmental factors such as temperature, habitat, and geographical location, which in turn affect their behavior and colonization patterns . The behavior of blowflies, including their approach patterns to carcasses, is another critical aspect of medico-legal entomology. Seasonal variations and environmental conditions significantly influence these behaviors, affecting the timing and nature of colonization .
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
39057269_p2
|
39057269
|
sec[0]/p[2]
|
1. Introduction
| 4.011719 |
biomedical
|
Study
|
[
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[
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0.00021910667419433594
] |
Localized studies have shown that regional variations significantly impact blowfly population dynamics, highlighting the importance of region-specific data in medico-legal entomology . Detailed studies on the ecological and biological aspects of blowfly activity can provide supplementary evidence in forensic cases, offering insights into the circumstances surrounding the discovery of remains . This is particularly relevant in regions like South Korea, where distinct seasonal and geographical factors can influence insect activity. While global databases like the Global Biodiversity Information Facility (GBIF) and the Korean Natural History Research Information System (NARIS) provide valuable biodiversity data, they often lack the regional and seasonal specificity needed for forensic applications . Therefore, there is a pressing need for comprehensive, quantitative data on blowfly species diversity and distribution across well-defined regions and time frames to effectively support forensic investigations.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p3
|
39057269
|
sec[0]/p[3]
|
1. Introduction
| 3.826172 |
biomedical
|
Study
|
[
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[
0.99951171875,
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0.00005137920379638672
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Globally, forensic research into the distribution and diversity of blowfly species is currently being conducted, with significant contributions from various countries, including the United States , Canada , Colombia , Argentina , Switzerland , Germany , Spain , and Japan . Despite these extensive efforts, research focusing on the forensically significant blowfly species in South Korea remains sparse, with a notable lack of comprehensive regional coverage. Thus, detailed investigations into the blowfly populations across various South Korean regions are required. In this study, we aim to explore the spatio-temporal distribution of forensically relevant blowflies in Gyeongsangnam-do, South Korea. Given the known impact of environmental factors such as temperature, habitat, and geographical location on blowfly species distribution and diversity, this research posits the hypothesis that community composition, species abundance, and diversity will differ across different regions, seasons, and habitats.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p4
|
39057269
|
sec[1]/sec[0]/p[0]
|
2.1. Site Description
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biomedical
|
Study
|
[
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[
0.990234375,
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This study encompasses a comprehensive survey conducted over the span of one year within four distinct regions of Gyeongsangnam-do, South Korea: Busan, Changwon, Gimhae, and Tongyeong . Each region was selected based on its unique geographical characteristics, as delineated below:
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p5
|
39057269
|
sec[1]/sec[0]/p[1]
|
2.1. Site Description
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other
|
Other
|
[
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[
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Busan (BS): This region is characterized by its mountainous terrain, is bordered by the sea, and includes islands along the eastern and southern coasts, conferring a diverse ecosystem.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p6
|
39057269
|
sec[1]/sec[0]/p[2]
|
2.1. Site Description
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other
|
Other
|
[
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[
0.003582000732421875,
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] |
Changwon (CW): Distinguished by a mountainous valley landscape, Changwon offers a unique combination of natural topographical features.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p7
|
39057269
|
sec[1]/sec[0]/p[3]
|
2.1. Site Description
| 1.047852 |
other
|
Other
|
[
0.077880859375,
0.0015172958374023438,
0.92041015625
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[
0.00506591796875,
0.994140625,
0.0004611015319824219,
0.00044608116149902344
] |
Gimhae (GH): This region is notable for its proximity to the South Sea and several rivers, providing a distinctive aquatic and terrestrial interface.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
39057269_p8
|
39057269
|
sec[1]/sec[0]/p[4]
|
2.1. Site Description
| 0.92627 |
other
|
Other
|
[
0.012420654296875,
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0.98681640625
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[
0.00482177734375,
0.994140625,
0.0006256103515625,
0.0006003379821777344
] |
Tongyeong (TY): Tongyeong is a peninsula known for its ria coast, and its landscape is marked by intricate coastlines and varied maritime habitats.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p9
|
39057269
|
sec[1]/sec[0]/p[5]
|
2.1. Site Description
| 3.154297 |
biomedical
|
Study
|
[
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[
0.9990234375,
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0.0000540614128112793
] |
To investigate the difference in species composition and abundance of blowflies based on habitat types within the study regions, the trap deployment sites were classified into two types: urban and forest. Urban habitats were defined by densely populated areas featuring man-made structures, such as roadsides or residential areas. Conversely, the forest habitats were characterized by sparsely populated areas, typically located at the base of mountains or within wooded terrains, devoid of man-made structures. The precise location of each trap was documented using GPS coordinates to ensure accuracy in data collection. The spatial distribution of the selected regions ranged from a minimum distance of approximately 15.3 km to a maximum distance of approximately 64.4 km, highlighting the study’s geographical breadth.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p10
|
39057269
|
sec[1]/sec[1]/p[0]
|
2.2. Sampling Procedure
| 3.066406 |
biomedical
|
Study
|
[
0.830078125,
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] |
[
0.99755859375,
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0.00008952617645263672
] |
Bait traps, recognized for their efficacy in capturing the broad characteristics of seasonal and regional variations in blowfly populations, were employed as the primary collection method . The survey was conducted twice a month in each region from May 2022 to May 2023. For each sampling event, a total of 10 bait traps were deployed: five in the urban habitats and five in the forest habitats. These traps were placed for a duration of 48 h, with an inter-trap distance of approximately 20–50 m, to ensure spatial independence . New traps were used for each survey to prevent any residual attractant effects. Thus, a total of 960 traps were deployed over the study period.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p11
|
39057269
|
sec[1]/sec[1]/p[1]
|
2.2. Sampling Procedure
| 1.933594 |
other
|
Study
|
[
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] |
[
0.6005859375,
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] |
The design of the bait trap consisted of a conical structure equipped with a hook at the apex, suspended 1.5–2 m above ground level . This elevation strategy was adopted to restrict access to ants and other non-target ground insects . As an attractant, thawed fresh small mouse carcasses weighing between 20 and 30 g were employed. These carcasses were brought in vacuum packed at room temperature before being used in the traps.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p12
|
39057269
|
sec[1]/sec[1]/p[2]
|
2.2. Sampling Procedure
| 2.679688 |
biomedical
|
Study
|
[
0.97021484375,
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[
0.88037109375,
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0.0005183219909667969
] |
Given the observed decrease in fly activity during periods of rainfall , efforts were made to schedule trap deployment during dry periods to avoid dilution of the attractant’s scent by rainwater, which may impede blowfly attraction. Upon collection, the blowflies were euthanized using a pyrethroid insecticide and preserved in 70% ethanol for subsequent analysis. Morphological identification of all adult specimens at the species level was conducted using the taxonomic keys provided by Ji and Kanō and Shinonaga . The specimens are preserved in the entomological collection at the Department of Biomedical Sciences, Kosin University, for future reference and study.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p13
|
39057269
|
sec[1]/sec[2]/p[0]
|
2.3. Data Analysis
| 3.285156 |
biomedical
|
Study
|
[
0.89013671875,
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] |
[
0.99755859375,
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0.0000820159912109375
] |
Blowfly community analysis was segmented by region, season, and habitat, with seasonal divisions according to the climatic patterns of Gyeongsangnam-do, South Korea, as delineated by the official weather website for spring (March to May), summer (June to August), autumn (September to November), and winter (December to February) . To elucidate the diversity of blowfly species in each season and region, metrics relative abundance, species diversity determined via the Shannon–Wiener index (H’), and species richness calculated through the Margalef index (R) were computed.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999994 |
39057269_p14
|
39057269
|
sec[1]/sec[2]/p[1]
|
2.3. Data Analysis
| 3.115234 |
biomedical
|
Study
|
[
0.986328125,
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] |
[
0.99267578125,
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0.00009804964065551758
] |
To reveal differences in temperature across regions and seasons, a one-way analysis of variance (ANOVA) was performed, followed by a Tukey’s HSD test as a post-hoc test. The average daily temperature was obtained from the Korea Meteorological Administration and was calculated from weather stations up to 10 km from the survey site.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p15
|
39057269
|
sec[1]/sec[2]/p[2]
|
2.3. Data Analysis
| 4.027344 |
biomedical
|
Study
|
[
0.99462890625,
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] |
[
0.9990234375,
0.00048160552978515625,
0.0002677440643310547,
0.00003927946090698242
] |
One-way ANOVA was employed to identify regional and seasonal disparities in species diversity and richness. Statistically significant differences were analyzed using post-hoc analysis with Tukey’s HSD test. Furthermore, the Kruskal–Wallis test was utilized to evaluate differences in relative abundance across seasons for the nine species that constituted more than 1% of the total community captured using the traps. Subsequent to significant Kruskal–Wallis test results, the Bonferroni correction was used to adjust the p -values obtained from Dunn’s test. Winter was excluded from the statistical analysis due to the zero variance of the data.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p16
|
39057269
|
sec[1]/sec[2]/p[3]
|
2.3. Data Analysis
| 3.994141 |
biomedical
|
Study
|
[
0.99609375,
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] |
[
0.9990234375,
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] |
The temporal dynamics of species abundance were visualized through a heatmap generated via two-way hierarchical clustering analysis, employing the unweighted pair group method with arithmetic mean (UPGMA) algorithm and Bray–Curtis similarity. Each month throughout the survey period was plotted along the X-axis, while the nine predominant blowfly species were presented along the Y-axis.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p17
|
39057269
|
sec[1]/sec[2]/p[4]
|
2.3. Data Analysis
| 4.058594 |
biomedical
|
Study
|
[
0.99609375,
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0.003330230712890625
] |
[
0.99951171875,
0.00020301342010498047,
0.0001971721649169922,
0.000032961368560791016
] |
Species diversity in different habitats was assessed using the Shannon–Wiener index (H’) and Margalef index (R), employing the same procedures as previously mentioned. Habitat preferences of the dominant species, representing more than 1% of the total community, were examined using a chi-square test of the trap data acquired throughout the study. To provide a more comprehensive view of the distribution patterns of blowflies, the relative abundances of environmental parameters (region and season, habitat) were analyzed using principal component analysis (PCA).
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p18
|
39057269
|
sec[1]/sec[2]/p[5]
|
2.3. Data Analysis
| 2.904297 |
biomedical
|
Study
|
[
0.99560546875,
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] |
[
0.873046875,
0.12493896484375,
0.0014142990112304688,
0.0004992485046386719
] |
For all statistical tests, the Shapiro–Wilk test was applied to assess data normality, and a p -value threshold of <0.05 was considered significant. Data analysis was performed in SPSS (version 23.0) and Python (version 3.12.0) along with the pandas (version 2.1.3), numpy (version 1.26.1), scipy (version 1.11.3), matplotlib (version 3.8.1), and seaborn (version 0.13.0) libraries.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
39057269_p19
|
39057269
|
sec[2]/sec[0]/p[0]
|
3.1. Seasonal and Regional Variations in Blowfly Abundances and Diversity
| 3.490234 |
biomedical
|
Study
|
[
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[
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0.00005745887756347656
] |
Throughout the investigation period, a total of 3470 adult blowflies, encompassing 13 species across five genera, were collected ( Table 1 ). Notably, no specimens were gathered during the winter season. Lucilia porphyrina emerged as the most prevalent species, with 1291 individuals accounting for 37.2% of the total collection, followed by Chrysomya pinguis (956 individuals representing 27.6%), Lucilia sericata (263 individuals, 7.6%), and Lucilia illustris (247 individuals, 7.1%).
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p20
|
39057269
|
sec[2]/sec[0]/sec[0]/p[0]
|
3.1.1. Analysis of Temperature Differences
| 3.757813 |
biomedical
|
Study
|
[
0.98828125,
0.0004374980926513672,
0.01146697998046875
] |
[
0.99951171875,
0.0003674030303955078,
0.00015747547149658203,
0.000038683414459228516
] |
The analysis of temperature differences between regions and seasons showed that there was no significant difference in the mean temperature between regions (F = 0.390, df = 3, p = 0.760), but there was a significant difference in the mean temperature between seasons . Furthermore, post-hoc analysis revealed that the mean temperature significantly differed between all pairs of seasons ( p < 0.001) .
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999994 |
39057269_p21
|
39057269
|
sec[2]/sec[0]/sec[1]/p[0]
|
3.1.2. Analysis of Blowfly Species Relative Abundance
| 4.117188 |
biomedical
|
Study
|
[
0.98876953125,
0.0005478858947753906,
0.0106658935546875
] |
[
0.99951171875,
0.00031256675720214844,
0.0003268718719482422,
0.00004112720489501953
] |
Analysis of the relative abundance of blowfly species across different regions and seasons revealed significant seasonal variations in species distribution . In the spring, Calliphora nigribarbis and L. porphyrina exhibited the highest abundance in Gimhae, constituting 22.1% of all blowflies, whereas only L. porphyrina was predominantly abundant in Busan, Changwon, and Tongyeong, with proportions of 43.7%, 37.4%, and 37.4%, respectively. During the summer months, Ch. pinguis exhibited the highest abundance in Gimhae and Changwon at 80.0% and 41.5%, respectively, while L. porphyrina was most prevalent in Busan and Tongyeong, accounting for 79.1% and 56.8%, respectively. In autumn, Calliphora calliphoroides was notably abundant in Gimhae, Changwon, and Tongyeong at 39.8%, 39.0%, and 34.6%, respectively, with L. porphyrina maintaining the highest abundance in Busan at 56.5%.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p22
|
39057269
|
sec[2]/sec[0]/sec[2]/p[0]
|
3.1.3. Analysis of Species Diversity
| 4.027344 |
biomedical
|
Study
|
[
0.9658203125,
0.0006051063537597656,
0.03363037109375
] |
[
0.99951171875,
0.0003437995910644531,
0.00022733211517333984,
0.000043272972106933594
] |
Although statistical analysis of species diversity indicated no significant differences between regions (F = 1.142, df = 3, p = 0.389), a notable variation between seasons (F = 4.817, df = 2, p = 0.038) was observed . Species diversity peaked in spring and was lowest in summer. Species diversity in autumn did not significantly differ from that in spring ( p = 0.398) or summer ( p = 0.246); however, there was a significant disparity in species diversity between spring and summer ( p = 0.031). Species richness showed no significant variation among regions (F = 1.749, df = 3, p = 0.234) or seasons (F = 0.873, df = 2, p = 0.450).
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
39057269_p23
|
39057269
|
sec[2]/sec[1]/p[0]
|
3.2. Seasonal Differences in Blowfly Abundance
| 4.109375 |
biomedical
|
Study
|
[
0.99609375,
0.00044608116149902344,
0.0035495758056640625
] |
[
0.99951171875,
0.0001615285873413086,
0.00016832351684570312,
0.00003796815872192383
] |
The Kruskal–Wallis test was applied to examine the seasonal relative abundances of the nine predominant captured blowfly species that constituted more than 1% of the total community ( Table 2 ). The results indicated significant seasonal variations in the abundances of four species. The abundance of C. calliphoroides , Calliphora grahami , and Ch. pinguis exhibited significant seasonal differences between summer and autumn ( p = 0.004, p = 0.043 and p = 0.013, respectively). The abundances of C. nigribarbis displayed notable seasonal differences between spring and summer ( p = 0.042).
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
39057269_p24
|
39057269
|
sec[2]/sec[1]/p[1]
|
3.2. Seasonal Differences in Blowfly Abundance
| 4.113281 |
biomedical
|
Study
|
[
0.99755859375,
0.0003733634948730469,
0.0018558502197265625
] |
[
0.99951171875,
0.000255584716796875,
0.00029754638671875,
0.00004214048385620117
] |
Two-way hierarchical clustering and heatmap analyses of the community composition of blowflies revealed distinct monthly correlations among the nine species . At the genus level, the clustering resulted in a discernible separation into three groups: one comprising four Calliphora species, one Chrysomya , and four Lucilia . Monthly clustering identified that March, April, October, and November, typically the early spring and late autumn months, formed distinct subgroups, while species in May through September clustered into another subgroup, indicating seasonal influences on community composition.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
39057269_p25
|
39057269
|
sec[2]/sec[2]/p[0]
|
3.3. Habitat Preferences and Environmental Influences on Blowfly Species Distribution
| 3.433594 |
biomedical
|
Study
|
[
0.9111328125,
0.0008339881896972656,
0.088134765625
] |
[
0.9990234375,
0.0007076263427734375,
0.00021767616271972656,
0.000057756900787353516
] |
The investigation into species diversity and richness across different habitats revealed that, in urban habitats, species diversity was 2.895 (H′), but was 2.019 (H′) in forest habitats, indicating a higher diversity in urban environments. Conversely, species richness presented similar values between habitats (1.153 (R) in urban habitats and 1.027 (R) in forest settings). These results suggest a distinction in habitat suitability for various blowfly species.
|
[
"Hyeon-Seok Oh",
"In-Seong Baek",
"Min-Gyu Kang",
"Sang-Hyun Park"
] |
https://doi.org/10.3390/insects15070536
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
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