New methods for drug synergy prediction: A mini-review.

In this mini-review, we explore the new prediction methods for drug combination synergy relying on high-throughput combinatorial screens. The fast progress of the field is witnessed in the more than thirty original machine learning methods published since 2021, a clear majority of them based on deep learning techniques. We aim to put these articles under a unifying lens by highlighting the core technologies, the data sources, the input data types and synergy scores used in the methods, as well as the prediction scenarios and evaluation protocols that the articles deal with. Our finding is that the best methods accurately solve the synergy prediction scenarios involving known drugs or cell lines while the scenarios involving new drugs or cell lines still fall short of an accurate prediction level.

Molecular typing and virulence characteristics of Escherichia coli strains isolated from hospital and community acquired urinary tract infections.

BACKGROUND: The main causes of hospital- and community-acquired urinary tract infections (UTIs) are a group of Escherichia coli (E. coli) strains with multiple virulence factors known as uropathogenic E. coli. METHODS AND RESULTS: One hundred E. coli isolates from the urine specimens of hospital- and community-acquired UTI patients were characterized based on their virulence factors and genetic relatedness using PCR and RAPD‒PCR, respectively. Among all, the traT (71%), sitA (64%), ompT (54%), malX (49%), ibeA (44%), tsh (39%), hlyD (18%) and cnf1 (12%) genes had the highest to lowest frequencies, respectively. There was no significant difference between the frequency of tested virulence genes in E. coli isolates from inpatients and outpatients. The frequency of the hlyD gene was significantly greater in E. coli isolates from patients hospitalized in gynecology, dermatology and intensive care unit (ICU) wards than in those from other wards. Eight virulence gene patterns were common among the isolates of inpatients in different wards of the same hospital, of which five patterns belonged to the isolates of inpatients in the same ward. More E. coli isolates with similar virulence gene patterns and greater genetic similarity were found in female patients than in male patients. The analysis of the RAPD‒PCR dendrograms revealed more genetic similarities among the E. coli isolates from inpatients than among those from outpatients. CONCLUSION: Our findings indicate the presence of a wide variety of virulence factors in E. coli isolates and the possibility of spreading the same clones in different wards of the hospital.

Direct additive-free N-formylation and N-acylation of anilines and synthesis of urea derivatives using green, efficient, and reusable deep eutectic solvent ([ChCl][ZnCl2]2).

In the current report, we introduce a simple, mild efficient and green protocol for N-formylation and N-acetylation of anilines using formamide, formic acid, and acetic acid as inexpensive, nontoxic, and easily available starting materials just with heating along stirring in [ChCl][ZnCl2]2 as a durable, reusable deep eutectic solvent (DES), which acts as a dual catalyst and solvent system to produce a wide range of formanilides and acetanilides. Also, a variety of unsymmetrical urea derivatives were synthesized by the reaction of phenyl isocyanate with a range of amine compounds using this benign DES in high to excellent yields. [ChCl][ZnCl2]2 showed good recycling and reusability up to four runs without considerable loss of its catalytic activity.

Severity of oxidative stress as a hallmark in COVID-19 patients.

INTRODUCTION: Understanding the mechanisms and identifying effective treatments for the COVID-19 outbreak are imperative. Therefore, this study aimed to assess the antioxidant status and oxidative stress parameters as potential pivotal mechanisms in asymptomatic, non-severe, and severe COVID-19 patients. METHODS: This study is a case-control study that was performed on patients referred to the Persian Gulf Martyrs Hospital of Bushehr University of Medical Sciences, Bushehr, Iran, from May 2021 to September 2021. A total of 600 COVID-19 patients (non-severe and severe group) and 150 healthy volunteers of the same age and sex were selected during the same period. On the first day of hospitalization, 10 ml of venous blood was taken from subjects. Then, hematological, biochemical, serological, antioxidant and oxidative stress parameters were determined. RESULTS: Our results indicated that ESR, CRP, AST, ALT, and LDH significantly augmented in the severe group as compared to the non-severe and normal groups (P ≤ 0.05). It was observed that the levels of FRAP, G6PD activity, and SOD activity significantly reduced in the non-severe patients in comparison with the severe and normal groups (P ≤ 0.05). We found that MDA content and NO metabolite markedly increased in severe patients as compared to the non-severe group. CONCLUSIONS: Taken together, it seems that the balance between antioxidants and oxidants was disturbed in COVID-19 patients in favor of oxidant markers. In addition, this situation caused more aggravation in severe patients as compared to the non-severe group.

Intraoral Extranodal Natural Killer Cell/T-Cell Lymphoma of the Hard Palate.

Natural killer cell/T-cell lymphoma (NK/TL) is a rare subtype of non-Hodgkin's lymphoma (NHL) associated with the Epstein-Barr virus (EBV) and requires early detection, which can be very challenging. NK/TL often arises in the nasal cavity and can then spread to the oral cavity, but the case presented here is an unusual presentation of NK/TL in a 66-year-old edentulous male patient who initially presented an intraoral exophytic lesion in the palate that appeared within 3 months. We report the present case to highlight the challenges of reaching an accurate diagnosis given the diverse clinical manifestations of NK/TL.

Effect of gamma-irradiated honey bee venom on blood parameter and histopathological observations of liver and kidney in a mice animal model.

Honey bee venom is a valuable product with a wide range of biological effects, whose use is rapidly increasing in apitherapy. In this study, the effect of gamma-irradiated honey bee venom (doses of 0, 2, 4, 6, and 8 kGy, volume of 0.1 ml, and concentration of 0.2 mg/ml) was evaluated on median lethal dose (LD50) determinations, liver and kidney histology, biochemical marker level, and serum protein analyses. Hence, the LD50 induced by the honey bee venom irradiated at 4, 6, and 8 kGy was increased, compared with the one at 0 and 2 kGy. Normal histology was observed in the liver and kidney of the mice receiving the honey bee venom irradiated at 4, 6, and 8 kGy. The serum levels of alanine aminotransferase (ALT) and all serum proteins were reduced at 4, 6, and 8 kGy compared with 0 and 2 kGy. Therefore, gamma irradiation at 4, 6, and 8 kGy had no negative effect on LD50, liver and kidney tissues, ALT, and serum protein levels by decreasing the allergen compounds of the honey bee venom.

Ion Release from Stainless Steel Brackets Subjected to Tooth Bleaching Treatment.

Statement of the Problem: Tooth discoloration in the form of staining is a common finding during conventional orthodontic treatment. Due to elevated esthetic standards, clinicians in the field of orthodontics are faced with increasing demands by patients to perform bleaching treatments while the appliances are still in place. Though the success of such treatments has been reported in literature, the effect of whitening agents on orthodontic appliances has not been evaluated. Increased ion release following corrosion of orthodontic brackets is considered a health hazard. Purpose: In this study, we measured the amount of ion release from steel brackets under home and office bleaching treatment in order to evaluate the safety of such treatments during orthodontic therapy. Materials and Method: In this experimental study, a total of 120 brackets were randomly divided into 3 groups. The first group was subjected to an office bleaching regimen (hydrogen peroxide 40%). The second group was subjected to a home bleaching treatment (carbamide peroxide 20%). The third group did not receive any bleaching treatment. The specimens of all groups were immersed in artificial saliva and after 30 days, the amount of released chromium, copper, iron, magnesium, and nickel ions was measured and compared using one way ANOVA. Results: The results showed that ion release was significantly different between the three groups (pValue >0.05). For the chromium, iron, magnesium, and Nickel ion release the order was as follows: no bleaching < home bleaching < office bleaching. However, the copper ion release was greater in the group that received home bleaching. Conclusion: Ion release was enhanced when bleaching treatments were performed, with office bleaching having a more significant effect. Although the amounts of released ions were less than those permitted by WHO, we suggest that the clinicians recommend home bleaching for orthodontic patients that are seeking tooth whitening treatment.

Iron Nanoparticles Wrapped with an N-Doped Carbon Material Produced by Using a Zinc Hydroxide Ferrocyanide Nanohybrid for Use in Commercial-like Supercapacitors.

According to the technology of carbon-based supercapacitors, modifying the structure of carbon as an active electrode material leads to an increase in capacitance. A modification involves introducing heteroatoms such as nitrogen into the carbon structure and composing it with metals such as iron. In this research, an anionic source called ferrocyanide was used to produce N-doped carbon consisting of iron nanoparticles. In fact, ferrocyanide was located as a guest between the layers of a host material, which is zinc hydroxide in the α phase. This new nanohybrid material was then heat-treated under Ar, and the heated product after acid washing was iron nanoparticles wrapped with N-doped carbon materials. This material was used as an active material in the production of symmetric supercapacitors with different organic (TEABF4 in acetonitrile) and aqueous (sodium sulfate) electrolytes as well as a new electrolyte (KCN in methanol). Accordingly, the supercapacitor made by the N/Fe-carbon active material and the organic electrolyte showed a capacitance value of 21 F/g at a current density of 0.1 A/g. This value is comparable to and even higher than the values observed in commercial supercapacitors.

Intravenous transplantation of bone marrow-derived mesenchymal stromal cells in patients with multiple sclerosis, a phase I/IIa, double blind, randomized controlled study.

Multiple sclerosis (MS) is a progressive, demyelinating neurodegenerative disease of the central nervous system. MS is immune-mediated and leads to disability especially in young adults. Even though 18 MS therapy drugs were approved, they slightly inhibit disease progression and do not induce regeneration and repair in the nervous system. Mesenchymal stromal cells (MSCs) have emerged as a new therapeutic modality in regenerative medicine and tissue engineering due to their immunomodulation and bio regenerative properties. We have designed a randomized, controlled clinical trial to assess safety and possible efficacy of MSC application in MS patients. Twenty-one MS patients were enrolled. Patients were allocated in two distinct groups: treatment group, which received systemic transplantation of autologous bone marrow-derived MSCs, and control group, which received placebo at the first injections. Patients in control group received MSCs at the second injection while the treatment group received placebo. All the patients were followed for 18 months. Follow-ups included regular visits, laboratory evaluation, and imaging analysis. Control patients received MSCs six month after treatment group. No severe immediate or late adverse events were observed in both groups after interventions. We did not find any significant differences in the rate of relapses, Expanded Disability Status Scale (EDSS) score, cognitive condition, Magnetic Resonance Imaging (MRI) findings, or any biomarkers of cerebrospinal fluid between the two groups and in each group before and after cell infusion. Transplantation of autologous bone marrow-derived mesenchymal stromal cells is safe and feasible. The efficacy of transplantation of these cells should be evaluated through designing randomized clinical trials with larger sample sizes, different administration routes, other cell types (allogeneic adipose derived MSCs, allogeneic Wharton's jelly derived MSCs …), repeated injections, and longer follow-up periods.

Evaluation and relation of oral health-related quality of life and oral health status in Thalassemia Major patients, a cross-sectional study.

BACKGROUND: Beta-Thalassemia is the most common human inherited disease, directly impacting patients' physical and psychosocial aspects. The present study evaluated oral health status, oral health-related quality of life (OHRQoL), and their correlation in Thalassemia Major patients. METHODS: Two hundred Thalassemia Major patients aged 12-49 participated in this cross-sectional study. Subjects were selected among the patients referred to the Faculty of Dentistry of Isfahan University of Medical Sciences using simple sampling. Oral Health Impact Profile 14 (OHIP 14) was used to assess OHRQoL. The decayed, Missing, and Filled Teeth (DMFT) index was used as an oral health index. SPSS 22 was used for analysis, using T and Pearson Correlation tests. RESULTS: 53% of participants were female, and 47% were male. The mean OHIP-14 score (± SD) was 13.20 (± 7.01). The mean DMFT score was 9.54 (± 5.72). DMFT and total OHIP 14 scores correlated significantly (p < 0.001, r = 0.78). All domains of the OHIP-14 score were also significantly correlated with DMFT (p < 0.05). CONCLUSIONS: Poor oral health conditions might adversely affect OHRQoL in TM patients. It seems necessary to provide oral treatment needs in order to improve OHRQoL in patients suffering from this particular disease.

Allogenic mesenchymal stromal cells and their extracellular vesicles in COVID-19 induced ARDS: a randomized controlled trial.

BACKGROUND AND AIMS: The main causes of death in patients with severe Coronavirus disease-2019 (COVID-19) are acute respiratory distress syndrome (ARDS) and multiorgan failure caused by a severe inflammatory cascade. Novel treatment strategies, such as stem-cell-based therapy and their derivatives can be used to relieve inflammation in these cases. In this study, we aimed to evaluate the safety and efficacy of therapy using mesenchymal stromal cells (MSCs) and their derived extracellular vesicles in COVID-19 patients. MATERIALS AND METHODS: COVID-19 patients with ARDS were included in this study and allocated into two study and control groups using block randomization. While all patients received recommended treatment based on guidelines from the national advisory committee for COVID-19 pandemic, the two intervention groups received two consecutive injections of MSCs (100 × 106 cells) or one dose of MSCs (100 × 106 cells) followed by one dose of MSC-derived extracellular vesicles (EVs). Patients were assessed for safety and efficacy by evaluating clinical symptoms, laboratory parameters, and inflammatory markers at baseline and 48 h after the second intervention. RESULTS: A total number of 43 patients (the MSC alone group = 11, MSC plus EV group = 8, and control group = 24) were included in the final analysis. Mortality was reported in three patients in the MSC alone group (RR: 0.49; 95% CI 0.14-1.11; P = 0.08); zero patient in the MSC plus EV group (RR: 0.08; 95% CI 0.005-1.26; P = 0.07) and eight patients in the control group. MSC infusion was associated with a decrease in inflammatory cytokines such as IL-6 (P = 0.015), TNF-α (P = 0.034), IFN-γ (P = 0.024), and CRP (P = 0.041). CONCLUSION: MSCs and their extracellular vesicles can significantly reduce the serum levels of inflammatory markers in COVID-19 patients, with no serious adverse events. Trial registration IRCT, IRCT registration number: IRCT20200217046526N2. Registered 13th April 2020, http://www.irct.ir/trial/47073 .

Predicting the Social-Emotional Competence Based on Childhood Trauma, Internalized Shame, Disability/Shame Scheme, Cognitive Flexibility, Distress Tolerance and Alexithymia in an Iranian Sample Using Bayesian Regression.

The purpose of this study was to predict Social Emotional Competence based on childhood trauma, internalized shame, disability/shame scheme, cognitive flexibility, distress tolerance, and alexithymia in an Iranian sample using Bayesian regression. The participants in this research were a sample of 326 (85.3% female and 14.7% male) people living in Tehran in 2021 who were selected by convenience sampling through online platforms. The survey assessments included demographic characteristics (age and gender), presence of childhood trauma, social-emotional competence, internalized shame, the Toronto Alexithymia scales, Young's measure of disability/shame together with measures of cognitive flexibility and distress tolerance. The results from Bayesian regression and Bayesian Model Averaging (BMA) indicated that internalized shame, cognitive flexibility and distress tolerance can be predictive of Social Emotional Competence. These results suggested that Social Emotional Competence can be explained by some important personality factors.

Design, synthesis, and biological evaluation of new 2-(4-(methylsulfonyl)phenyl)-N-phenylimidazo[1,2-a]pyridin-3-amine as selective COX-2 inhibitors.

Cyclooxygenase (COX), which plays a role in converting arachidonic acid to inflammatory mediators, could be inhibited by non-steroidal anti-inflammatory drugs (NSAIDs). Although potent NSAIDs are available for the treatment of pain, fever, and inflammation, some side effects, such as gastrointestinal ulcers, limit the use of these medications. In recent years, selective COX-2 inhibitors with a lower incidence of adverse effects attained an important position in medicinal chemistry. In order to introduce some new potent COX-2 inhibitors, a new series of 2-(4-(methylsulfonyl)phenyl)-N-phenylimidazo[1,2-a]pyridin-3-amines was designed, synthesized, and evaluated. The docking studies performed by AutoDock Vina demonstrated that docked molecules were positioned as well as a crystallographic ligand in the COX-2 active site, and SO2Me pharmacophore was inserted into the secondary pocket of COX-2 and formed hydrogen bonds with the active site. The designed compounds were synthesized through two-step reactions. In the first step, different 1-(4-(methylsulfonyl)phenyl)-2-(phenylamino)ethan-1-one derivatives were obtained by the reaction of aniline derivatives and α-bromo-4-(methylsulfonyl)acetophenone. Then, condensation of intermediates with different 2-aminopyridines gave final compounds. Enzyme inhibition assay and formalin test were performed to evaluate the activity of these compounds. Among these compounds, 8-methyl-2-(4-(methylsulfonyl)phenyl)-N-(p-tolyl)imidazo[1,2-a]pyridin-3-amine (5n) exhibited the highest potency (IC50 = 0.07 µM) and selectivity (selectivity index = 508.6) against COX-2 enzyme (selectivity index: COX-1 IC50/COX-2 IC50). The antinociceptive activity assessment via the formalin test showed that nine derivatives (5a, 5d, 5h, 5i, 5k, 5q, 5r, 5s, and 5t) possessed significant activity compared with the control group with a p value less than 0.05.

Effect of gamma-irradiated honey bee venom on gene expression of inflammatory and anti-inflammatory cytokines in mice.

In this study, the effect of gamma-irradiated honey bee venom (doses of 0, 2, 4, 6, and 8 kGy, volume of 0.1 ml and concentration of 0.2 mg/ml) was evaluated on the reduction of allergen compounds and the gene expression of inflammatory and anti-inflammatory cytokines in mice. Hence, edema activity induced by the bee venom irradiated at 4, 6, and 8 kGy was reduced, compared with the control group and that irradiated at 2 kGy. In contrast, the paw edema induced by the bee venom irradiated at 8 kGy increased, compared with 4 and 6 kGy. At all the time periods, there was a significant decrease in the gene expression of interferon gamma (IFN-γ), interleukin 6 (IL-6), and interleukin 10 (IL-10) in the bee venoms irradiated at 4, 6, and 8 kGy, compared with the control group and that irradiated at 2 kGy. In contrast, there was an increase in the gene expression of IFN-γ and IL-6 in the bee venom irradiated at 8 kGy, compared with those irradiated at 4 and 6 kGy. Therefore, gamma irradiation at 4 and 6 kGy reduced the gene expression of cytokines at each time period by decreasing the allergen compounds of honey bee venom.

The effect of sage (Salvizan gel) compared to triamcinolone acetonide on the treatment of recurrent aphthous stomatitis: a double-blinded randomized clinical trial.

BACKGROUND: Recurrent Aphthous Stomatitis (RAS) is one of the most common lesions of the oral mucosa. Herbal medicine can be used for the treatment of this disease. The present study aimed to compare the effects of topical application of Sage (Salvizan) and triamcinolone acetonide gels on RAS. METHODS: This double-blind clinical study recruited sixty patients with minor aphthous ulcers. Half of the patients were treated with Salvizan gel, and the other half were treated with oral triamcinolone acetonide gel. The effect of Salvizan topical gel was evaluated and compared with that of oral triamcinolone acetonide gel. Factors such as Pain recovery time, wound healing time, and pain level was evaluated. Data were analyzed by SPSS version 22 using independent t-test, paired t-test, repeated measures ANOVA, and survival analysis, including Kaplan-Meier and Cox regression. RESULTS: The mean duration of pain recovery was 1.5 days for Salvizan and 2.5 days for triamcinolone acetonide (p < 0.001). Moreover, the duration of wound healing was 3.3 days for Salvizan and 6 days for triamcinolone acetonide (p < 0.001). Patients' satisfaction from factors such as taste and smell had no significant difference between the two groups. CONCLUSION: The results of this study showed that Salvizan gel is very effective in the treatment of RAS. It was significantly better than triamcinolone acetonide in the pain recovery and wound healing. These promising results favor herbal treatments and show that they can be used more commonly for treating diseases such as RAS. TRIAL REGISTRATION: This study was approved by the ethics committee of Isfahan University of Medical Sciences (IR.MUI. RESEARCH: REC.1399.834) on 17/03/2021. It was also registered in the Iranian Registry of Clinical Trials (IRCT20100202003251N7).

A survival analysis of socio-demographic and clinical predictors among hospitalized COVID-19 patients in Southern Iran.

BACKGROUND: This study aimed to evaluate the socio-demographic, clinical, and laboratory risk factors in hospitalized COVID-19 patients during the first 6 months of the SARS-CoV-2 epidemic. METHOD: This retrospective hospital-based cross-sectional study included all laboratory-confirmed cases of the COVID-19 virus that were admitted to the Shohadaye-Khalije-Fars Hospital in Bushehr, Iran, from February 22, 2020 to September 21, 2020. The patients' records were reviewed during the hospitalization period. The global COVID-19 clinical platform, i.e., the World Health Organization Rapid Case Report Form was used as the data collection tool. We conducted the survival analysis using the Kaplan-Meier and the Stepwise Cox regression analyses. RESULTS: The analysis included 2108 confirmed cases of COVID-19 with a mean age of 47.81 years (SD 17.78); 56.8% men, 43.2% women and 6.3% (n = 133) deaths. After adjustment, it was found that factors associated with an increased risk of death consisted of chronic kidney disease, intensive care unit admission, cancer, and hemoptysis. The 7-day survival rate was 95.8%, which decreased to 95.1%, 94.0%, and 93.8% on days 14, 21, and 28 of hospitalization, respectively. DISCUSSION AND CONCLUSION: Older COVID-19 patients with manifestation of hemoptysis and a past medical history of chronic kidney disease and cancer, should be closely monitored to prevent disease deterioration and death, and also should be admitted to the intensive care unit.

Evaluation of last-year dental students' knowledge of prescription of analgesics and anti-bacterial agents for pregnant and lactating women

Oral health in pregnant and lactating women can affect maternal and child health. Dental treatments in this period should not have adverse effects on maternal and child health. This study was conducted to investigate the last-year dental students' knowledge of the prescription of analgesics and antibiotics for pregnant and lactating mothers in Isfahan city, Iran. Methods: A total of 104 last-year dental students of Khorasgan Islamic Azad University and Isfahan University of Medical Sciences were recruited in this descriptiveanalytical study. They completed a 16-item questionnaire on the prescription of analgesics and antibiotics for pregnant and lactating mothers. Data were analyzed by SPSS (Version 22) statistical software(t-test). Results: The total mean score of students' knowledge was 9.02±1.91. The highest level of knowledge about the safest analgesic (acetaminophen) was reported for the pregnant patients so that 100% of them had correct information in this regard. Further, the lowest level of knowledge was found for the use of dexamethasone during lactation, as only 10.6% of respondents were able to provide a correct response in this regard. Conclusion: The dental students had average knowledge about the prescription of antibiotics and analgesics for pregnant and lactating patients. To prevent the possible risks for these patients, further information should be provided to dental students

Oral health-related quality of life and orthodontic treatment need in thalassemia major patients.

Background: Thalassemia major (TM) is a severe life-threatening hemoglobinopathy. It causes a typical chipmunk face due to increased hematopoiesis. Severe malocclusion often accompanies facial deformity, which may affect Oral Health-related Quality of Life (OHRQoL). The aim of this study was to assess the relationship between orthodontic treatment needs and OHRQoL in TM patients. Materials and Methods: One hundred and five adult patients with TM participated in this cross-sectional study. Oral Health Impact Profile-14 (OHIP-14) questionnaire was used to measure OHRQoL and the patient's need for orthodontic treatment was evaluated using the Index of Orthodontic Treatment Need (IOTN). The comparison of OHIP-14 scores between groups regarding orthodontic treatment need was carried out using t-test (SPSS software); P < 0.05 was considered statistically significant. Results: The mean age of participants was 24.92 (±9.33) years, with 52% female versus 48% male. Orthodontic treatment need was 27.6%. The mean score of OHIP-14 was 12.95 (±7.02). A statistically significant relationship was found between OHIP-14 score and age, gender, and orthodontic treatment need (P < 0.05). All domains of OHIP-14 were significantly related to orthodontic treatment need (P < 0.05), except for "physical disability" (P = 0.282). Conclusion: OHRQoL was lower in TM patients with orthodontic treatment needs. Planning to treat malocclusion seems necessary to improve the quality of life in these patients.

Analysis of monocyte cell tractions in 2.5D reveals mesoscale mechanics of podosomes during substrate-indenting cell protrusion.

Podosomes are mechanosensitive protrusive actin structures that are prominent in myeloid cells, and they have been linked to vascular extravasation. Recent studies have suggested that podosomes are hierarchically organized and have coordinated dynamics on the cell scale, which implies that the local force generation by single podosomes can be different from their global combined action. Complementary to previous studies focusing on individual podosomes, here we investigated the cell-wide force generation of podosome-bearing ER-Hoxb8 monocytes. We found that the occurrence of focal tractions accompanied by a cell-wide substrate indentation cannot be explained by summing the forces of single podosomes. Instead, our findings suggest that superimposed contraction on the cell scale gives rise to a buckling mechanism that can explain the measured cell-scale indentation. Specifically, the actomyosin network contraction causes peripheral in-plane substrate tractions, while the accumulated internal stress results in out-of-plane deformation in the central cell region via a buckling instability, producing the cell-scale indentation. Hence, we propose that contraction of the actomyosin network, which connects the podosomes, leads to a substrate indentation that acts in addition to the protrusion forces of individual podosomes. This article has an associated First Person interview with the first author of the paper.

A Roadmap for the Production of a GMP-Compatible Cell Bank of Allogeneic Bone Marrow-Derived Clonal Mesenchymal Stromal Cells for Cell Therapy Applications.

BACKGROUND: Allogeneic mesenchymal stromal cells (MSCs) have been used extensively in various clinical trials. Nevertheless, there are concerns about their efficacy, attributed mainly to the heterogeneity of the applied populations. Therefore, producing a consistent population of MSCs is crucial to improve their therapeutic efficacy. This study presents a good manufacturing practice (GMP)-compatible and cost-effective protocol for manufacturing, banking, and lot-release of a homogeneous population of human bone marrow-derived clonal MSCs (cMSCs). METHODS: Here, cMSCs were isolated based on the subfractionation culturing method. Afterward, isolated clones that could reproduce up to passage three were stored as the seed stock. To select proliferative clones, we used an innovative, cost-effective screening strategy based on lengthy serial passaging. Finally, the selected clones re-cultured from the seed stock to establish the following four-tired cell banking system: initial, master, working, and end of product cell banks (ICB, MCB, WCB, and EoPCB). RESULTS: Through a rigorous screening strategy, three clones were selected from a total of 21 clones that were stored during the clonal isolation process. The selected clones met the identity, quality, and safety assessments criteria. The validated clones were stored in the four-tiered cell bank system under GMP conditions, and certificates of analysis were provided for the three-individual ready-to-release batches. Finally, a stability study validated the EoPCB, release, and transport process of the frozen final products. CONCLUSION: Collectively, this study presents a technical and translational overview of a GMP-compatible cMSCs manufacturing technology that could lead to the development of similar products for potential therapeutic applications.