Potential therapeutic effects and nano-based delivery systems of mesenchymal stem cells and their isolated exosomes to alleviate acute respiratory distress syndrome caused by COVID-19.

The severe respiratory effects of the coronavirus disease 2019 (COVID-19) pandemic have necessitated the immediate development of novel treatments. The majority of COVID-19-related fatalities are due to acute respiratory distress syndrome (ARDS). Consequently, this virus causes massive and aberrant inflammatory conditions, which must be promptly managed. Severe respiratory disorders, notably ARDS and acute lung injury (ALI), may be treated safely and effectively using cell-based treatments, mostly employing mesenchymal stem cells (MSCs). Since the high potential of these cells was identified, a great deal of research has been conducted on their use in regenerative medicine and complementary medicine. Multiple investigations have demonstrated that MSCs and their products, especially exosomes, inhibit inflammation. Exosomes serve a critical function in intercellular communication by transporting molecular cargo from donor cells to receiver cells. MSCs and their derived exosomes (MSCs/MSC-exosomes) may improve lung permeability, microbial and alveolar fluid clearance, and epithelial and endothelial repair, according to recent studies. This review focuses on COVID-19-related ARDS clinical studies involving MSCs/MSC-exosomes. We also investigated the utilization of Nano-delivery strategies for MSCs/MSC-exosomes and anti-inflammatory agents to enhance COVID-19 treatment.

Osteogenic Differentiation of Adipose Tissue-Derived Mesenchymal Stem Cells on Composite Polymeric Scaffolds: A Review.

The mesenchymal stem cells (MSCs) are the fundamental part of bone tissue engineering for the emergence of reconstructive medicine. Bone tissue engineering has recently been considered a promising strategy for treating bone diseases and disorders. The technique needs a scaffold to provide an environment for cell attachment to maintain cell function and a rich source of stem cells combined with appropriate growth factors. MSCs can be isolated from adipose tissue (ASCs), bone marrow (BM-MSCs), or umbilical cord (UC-MSCs). In the present study, the potential of ASCs to stimulate bone formation in composite polymeric scaffolds was discussed and it showed that ASCs have osteogenic ability in vitro. The results also indicated that the ASCs have the potential for rapid growth, easier adipose tissue harvesting with fewer donor site complications and high proliferative capacity. The osteogenic differentiation capacity of ASCs varies due to the culture medium and the addition of factors that can change signaling pathways to increase bone differentiation. Furthermore, gene expression analysis has a significant impact on improving our understanding of the molecular pathways involved in ASCs and, thus, osteogenic differentiation. Adding some drugs, such as dexamethasone, to the biomaterial composite also increases the formation of osteocytes. Combining ASCs with scaffolds synthesized from natural and synthetic polymers seems to be an effective strategy for bone regeneration. Applying exopolysaccharides, such as schizophyllan, chitosan, gelatin, and alginate in composite scaffolds enhances the osteogenesis potential of ASCs in bone tissue regeneration.

Prophylaxis against intraocular pressure spikes following uncomplicated phacoemulsification: a systematic-review and meta-analysis.

BACKGROUND: To investigate the effect of perioperative intraocular pressure (IOP) lowering medications on controlling postoperative IOP following uncomplicated phacoemulsification. METHODS: Ovid MEDLINE, EMBASE, and Cochrane CENTRAL databases were searched up until November 2022. Randomised controlled trials (RCTs) that assessed IOP change via applanation tonometry in medicated and control arms following uncomplicated cataract surgery in healthy eyes were included. The primary outcome was the weighted mean difference (WMD) of IOP at 2-8 h, 12-24 h, and 1-7 days postoperatively within each medication class or common fixed-combination formulations. Risk of bias was assessed using the revised risk of bias in randomised trials (RoB-2). Level of evidence was rated using the Grading of Recommendation, Assessment, Development and Evaluation (GRADE) RESULTS: From 702 screened articles, 30 RCTs involving 2986 eyes were included. There was a statistically significant reduction in IOP favouring treatment arms at 2-8 h (WMD = -3.87 mmHg; 95% CI [-4.75, -3.00]; p < 0.001) and 12-24 h (WMD = -2.69 mmHg; 95% CI [-3.36, -2.02]; p < 0.001), with the effect wearing off beyond 1 day (p = 0.18). Between medication classes, the largest effect at both 2-8 h and 12-24 h was observed with intracameral cholinergics or fixed-combination carbonic anhydrase inhibitor-beta-blocker (FCCB) formulations. Conversely, the smallest effect was observed with prostaglandin analogues, alpha-agonists, and topical carbonic anhydrase inhibitors (CAIs). CONCLUSION: Prophylaxis against acute IOP elevations following uncomplicated cataract surgery is effective. FCCB and intracameral cholinergics are the most effective ocular antihypertensive agents, while alpha-agonists, prostaglandin analogues, and topical CAIs were found to be the least effective. These findings may inform future surgical guidelines.

Canadian legal cases in ophthalmology: a closer look at 40 years' worth of data.

OBJECTIVE: To assess the characteristics of malpractice lawsuits against Canadian ophthalmologists and the predisposing factors leading to claims. DESIGN: Retrospective case series. METHODS: A systematic search of the 2 largest Canadian online legal databases, LexisNexis Canada and Westlaw Canada, was performed to collect cases against ophthalmologists in Canadian courts from 1977 to 2021. RESULTS: This study comprised 68 legal cases, including 52 lawsuits, 14 cases appealed once, and 2 cases appealed twice. Most cases concerned surgical procedures (46.2%), followed by misdiagnoses or lack thereof (32.7%) and nonsurgical procedures (21.2%). Half the cases (n = 26) were immediately dismissed by the judge in favour of the ophthalmologist, though among the remaining half that proceeded to trial the majority (88.5%) were won by the patients. All appeals by patients were dismissed by the judge. The median monetary value of damages awarded was $308,202. CONCLUSIONS: This study is the first to report on ophthalmology-involved medical litigation cases in Canada. Most cases were ruled in favour of the ophthalmologist, but most of those that were not immediately dismissed by the judge were ruled in favour of the plaintiff. Notably, a plurality of these cases argued for a lack of informed consent, and every case in which a lack was successfully pleaded was ruled in favour of the plaintiff, highlighting the importance of appropriate informed consent. The findings of this study give Canadian ophthalmologists insight into areas of practice that commonly lead to litigation and can aid in improving clinical practice and risk management.

Mesenchymal stem cells osteogenic differentiation by ZnO nanoparticles and polyurethane bimodal foam nanocomposites.

Mesenchymal stem cells with tissue repair capacity involve in regenerative medicine. MSCs can promote bone repair when employed with nano scaffolds/particles. Here, the MTT and Acridine Orange assay enabled the cytotoxic concentration of Zinc oxide nanoparticles and Polyurethane evaluation. Following culturing adipose tissue-derived MSCs, ADSCs' proliferation, growth, and osteogenic differentiation in the presence of PU with and without ZnO NPs is tracked by a series of biological assays, including Alkaline Phosphatase activity, Calcium deposition, alizarin red staining, RT-PCR, scanning electron microscope, and immunohistochemistry. The results showed boosted osteogenic differentiation of ADSCs in the presence of 1% PU scaffold and ZnO NPS and can thus apply as a new bone tissue engineering matrix. The expression level of Osteonectin, Osteocalcin, and Col1 increased in PU-ZnO 1% on the 7th and 14th days. There was an increase in the Runx2 gene expression on the 7th day of differentiation in PU-ZnO 1%, while it decreased on day 14th. In conclusion, Polyurethane nano scaffolds supported the MSCs' growth and rapid osteogenic differentiation. The PU-ZnO helps not only with cellular adhesion and proliferation but also with osteogenic differentiation.

Schizophyllan promotes osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells in vitro.

BACKGROUND: Bioactive polysaccharides are a promising way for bone disease prevention with high efficiency. Schizophyllan (SPG) is a polysaccharide derived from a species of fungus with anticancer, antitumor, and anti-inflammatory effects. In the present study, for the first time, the cell proliferation, osteogenic markers, mineral deposition, and osteogenic gene expression of human adipose tissue-derived mesenchymal stem cells (hADMSCs) grown on SPG were evaluated by in vitro assays. METHODS AND RESULTS: The cytotoxicity of SPG was measured using the MTT assay and acridine orange staining. Differentiation of hADMSCs was assessed using alkaline phosphatase (ALP) activity test, cellular calcium content assay, and mineralized matrix staining. To this end, Alizarin red S, von Kossa staining, and the expression of bone-specific markers, including ALP, Runx2, and osteonectin, were used by real-time RT-PCR over a 2-week period. According to the results, SPG at 10 µg/ml concentration was determined as the optimal dosage for differentiation studies. The results of osteogenic differentiation tests showed that compared to the control groups in vitro, SPG enhanced the osteogenic markers and mineralization as well as upregulation of the expression of bone specific genes in differentiated hADMSCs during differentiation. CONCLUSIONS: The results revealed that SPG could be applied as effective factor for osteogenic differentiation in the future. The current study provides insights into the hADMSC-based treatment and introduces promising therapeutic material for individuals who suffer from bone defects and injuries.

Apoptotic effects of mesenchymal stem cells' conditioned medium on colorectal cancer cell lines.

Multipotent Mesenchymal stem cells (MSCs) have vigorous immunomodulatory activity, apoptotic effects, and the capacity to migrate to inflammatory and tumor sites. This study focuses on the apoptotic effects of MSCs conditioned medium (CM) on colorectal cancer cell lines. MSCs were preconditioned with lipopolysaccharide (LPS) to induce apoptosis in colorectal cancer cells. The conditioned medium (LPS-CM) from the preconditioned cells was isolated and used to treat colorectal cancer cells (HT29 and SW48). The survival and proliferation of cancer cells were assessed using Trypan blue staining and MTT assay. The apoptosis rate was evaluated through flow cytometry analysis and caspase-3 activity. Additionally, Real-Time PCR was used to measure the mRNA level of apoptotic and anti-apoptotic factors, including bcl2, bax, and p53 genes. The results showed that LPS-CM significantly increased (p < 0.001) the percentage of apoptosis in the SW48 and HT29 cell lines. Caspase-3 activity significantly increased (p < 0.001) in these cell lines after treatment with LPS-CM. The mRNA level of bcl2 was significantly decreased (p < 0.001), while bax and p53 genes were significantly overexpressed (p < 0.001) in the LPS-CM treated cell lines. Notably, the mRNA level of bcl2 and bax genes was significantly altered at a higher concentration of LPS-CM. In conclusion, the conditioned medium from LPS-preconditioned MSCs can effectively induce apoptosis in colorectal cancer cells. This finding suggests that LPS-CM could be a potential strategy for inhibiting the proliferation and progression of colorectal cancer cells.

Gender Equality in Plastic Surgery Training: A Canadian Nationwide Cross-sectional Analysis.

Introduction: One of the important factors in achieving gender equity is ensuring equitable surgical training for all. Previous studies have shown that females get significantly lower surgical exposure than males in certain surgical specialties. Gender gap in surgical exposure has never been assessed in plastic surgery. To that end, the goal of this study was to assess if there are any differences in plastic surgery training between male and female residents. Methods: A survey was sent to all plastic surgery residency programs in Canada to assess the No. of surgeries residents operated on as a co-surgeon or primary assistant during their training. The survey also assessed career goals, level of interest in the specialty, and subjective perception of gender bias. Results: A total of 89 plastic surgery residents (59.3% participation rate) completed the survey and were included in the study. The average No. of reconstructive cases residents operated on as a co-surgeon or primary assistant was 245 ± 312 cases. There was no difference in either reconstructive or aesthetic surgery case logs between male and female residents (p > .05). However, a significantly larger proportion of females (39%) compared to males (4%) felt that their gender limited their exposure to surgical cases and led to a worsening of their overall surgical training (p < .001). Finally, a larger proportion of male residents were interested in academic careers while a larger proportion of female residents were interested in a community practice (p = .024). Conclusion: While there is no evidence of differences in the volume of logged cases between genders, female surgical residents still feel that their respective gender limits their overall surgical training. Gender inequalities in training should be addressed by residency programs.

Introduction: L'un des facteurs importants pour atteindre l'égalité des genres est d'assurer une formation chirurgicale équitable pour tous. Des études antérieures ont montré que les femmes ont une exposition significativement moindre à la chirurgie que les hommes dans certaines spécialités chirurgicales. L'écart entre genres pour l'exposition à la chirurgie n'a jamais été évalué en chirurgie plastique. À cette fin, la présente étude a eu pour objectif d'évaluer s'il y avait des différences dans la formation à la chirurgie plastique entre les résidents masculins et féminins. Méthodes: Une enquête a été envoyée à tous les programmes canadiens de résidence en chirurgie plastique pour évaluer le nombre d'interventions auxquelles les résidents ont participé en tant que co-chirurgien ou assistant principal au cours de leur formation. L'enquête a également évalué les objectifs de carrière, le niveau d'intérêt dans la spécialité et la perception subjective d'un biais lié au genre. Résultats: En tout, 89 résidents en chirurgie plastique (taux de participation de 59,3 %) ont répondu à l'enquête et ont été inclus dans l'étude. Le nombre moyen de cas de chirurgie reconstructrice au cours desquelles les résidents sont intervenus en tant que co-chirurgien ou principal assistant était de 245 ± 312 cas. Il n'y a pas eu de différence entre les journaux de cas, qu'il s'agisse de chirurgie reconstructrice ou de chirurgie esthétique entre résidents masculins et féminins (P > 0,05). Cependant, un nettement plus grand pourcentage de femmes (39 %) que d'hommes (4 %) estimait que leur genre limitait leur exposition à des cas chirurgicaux et résultait dans une aggravation de leur formation globale à la chirurgie (P < 0,001). Enfin, un plus grand pourcentage de résidents masculins était intéressé par une carrière universitaire alors qu'un plus grand pourcentage de résidentes était intéressé par une pratique dans la communauté (P = 0,024). Conclusion: Bien qu'il n'y ait pas de données probantes étayant des différences de volume des cas consignés entre les genres, les résidentes féminines en chirurgie pensent encore que leur genre limite leur formation chirurgicale. Les inégalités entre genres devraient être abordées par les programmes de résidence.

Outcomes and risk factors for Kahook Dual Blade excisional goniotomy with concomitant phacoemulsification: a multicentre Canadian study.

OBJECTIVE: To assess the outcomes and failure risk factors for Kahook Dual Blade (KDB) excisional goniotomy with cataract surgery (phaco-KDB) in eyes with various glaucoma subtypes and severities. METHODS: This multisurgeon consecutive case series included glaucomatous eyes with cataract that underwent phaco-KDB and had a minimum follow-up of 12 months postoperatively. Efficacy was assessed by absolute and qualified surgical success (defined by different criteria) and changes in intraocular pressure (IOP) and antiglaucoma medication (AGM) at the last postoperative follow-up. Safety included best-corrected visual acuity, cup-to-disc ratio, visual field mean deviation, retinal nerve fibre layer thickness, and adverse events. RESULTS: A total of 108 eyes of 89 patients with a median follow-up of 18 months (range, 12-47 months) were included. IOP decreased by 26% from 19.1 ± 5.0 mm Hg to 14.1 ± 3.5 mm Hg (p < 0.001), AGM use decreased by 29% from 2.4 ± 1.3 medications to 1.7 ± 1.3 (p < 0.001), and 25% of eyes became free of AGMs (vs 3% at baseline). Qualified success rates achieved for IOP cutoffs of 18, 15, and 12 mm Hg were 87%, 68%, and 46%, respectively. Higher baseline IOP and postoperative incidence of IOP spikes were associated with a higher risk of surgical failure. Best-corrected visual acuity improved postoperatively (p < 0.001), and visual field mean deviation, cup-to-disc ratio, and retinal nerve fibre layer thickness remained stable. Overall, safety was favourable, and adverse events were transient and not sight threatening. CONCLUSION: This multicentre Canadian study provides real-world data that support the safety and efficacy of phaco-KDB in reducing IOP and AGM use with no evidence of disease progression during the follow-up period.

Combination of natural scaffolds and conditional medium to induce the differentiation of adipose-derived mesenchymal stem cells into keratocyte-like cells and its safety evaluation in the animal cornea.

Keratocytes are the main cellular components of the corneal stroma. This cell is quiescent and cannot be cultured easily. The aim of this study was to investigate differentiate human adipose mesenchymal stem cells (hADSCs) into corneal keratocyte cells by combining natural scaffolds and conditioned medium (CM) and evaluating their safety in the rabbit's cornea. Keratocytes were cultured in an optimal culture medium and this medium was collected and kept as a CM. hADSCs were cultured on the decellularized human small incision lenticule extraction (SMILE) lenticule (SL), amniotic membrane (AM), and collagen-coated plates, and were exposed to keratocyte-CM (KCM) for 7, 14, and 21 days. Differentiation was evaluated using Real-time PCR and immunocytochemistry (ICC). hADSCs were cultured on the SL scaffolds and implanted in the corneal stroma of 8 New Zealand male rabbits. Rabbits were followed for 3 months and the safety was evaluated by clinical and histological variables. Real-time PCR results showed a significant increase in the expression of keratocyte-specific markers on the 21 day of differentiation compared to the control group. ICC also confirmed the induction of differentiation. Implantation of SLs containing differentiated cells in the cornea of animals showed no serious complications including neovascularization, corneal opacity, inflammation, or signs of tissue rejection. Furthermore, the evaluation of the presence of keratocyte-like cells after three months in the rabbit stroma was confirmed by Real-time PCR and immunohistochemistry (IHC) analysis. Our results showed that combination of combination of corneal extracellular matrix and KCM can induced keratocytes differentiation of hADSC and can be introduced as a alternative method to supply the required keratocytes in corneal tissue engineering.

Large Language Models in Ophthalmology Scientific Writing: Ethical Considerations Blurred Lines or Not at All?

PURPOSE: To discuss the implications of large language models (LLMs) in ophthalmology research, as well as the associated ethical considerations. DESIGN: Perspective. METHODS: This discussion reviews the potential uses of LLMs such as ChatGPT in ophthalmology research, highlights the associated threats and ethical considerations, and proposes solutions for the use of LLMs in ophthalmology research and scientific writing. RESULTS: With the increasing interest in LLMs, such as ChatGPT, their diverse utility has been widely explored, including their application in research and scientific writing. LLMs have the potential to guide researchers throughout the different stages of their research, from idea generation to drafting a scientific piece. However, there are significant ethical concerns and challenges related to scientific integrity in ophthalmology research that should be addressed by scientific journals. Our review of the 10 highest-impact-factor ophthalmology journals revealed that the number of journals addressing this topic in their submission guidelines is rapidly increasing. Therefore, we propose certain domains that all journals should consider regarding the use of LLMs in research. CONCLUSIONS: As LLMs continue to improve, their use in scientific writing will remain a contentious issue due to the ethical dilemmas involved in determining the appropriate scope of their use. This article reviews the ethical dilemmas related to the use of LLMs in ophthalmology research and calls for the prompt development of guidelines for their ethical use in manuscript writing as ophthalmology journals update their editorial policies with respect to LLMs.

Retinal vein occlusion in patients with primary angle closure: a prospective case-control study.

OBJECTIVE: To compare the prevalence of angle closure in eyes with retinal vein occlusion (RVO) with control eyes and assess the possible association between angle-closure and RVO. PATIENTS AND METHODS: This prospective, blinded case-control study included patients with a history of RVO (cases) and control individuals matched for age and refractive error. Clinical characteristics and angle-based structures derived from anterior-segment optical coherence tomography (AS-OCT) were analyzed. RESULTS: Eighty-eight patients (44 per group) were included. The average age of the RVO and control groups was 59.8 ± 11.6 years and 60.8 ± 9.0 years, respectively (p = 0.667). There were no significant differences in terms of clinical characteristics between the 2 groups, including intraocular pressure (p = 0.837) and Shaffer gonioscopy grading (p = 0.620). None of the AS-OCT-derived angle characteristics were significantly different between the 2 groups. The number of angle-closure diagnoses between the RVO group (1 primary angle closure and 7 primary angle-closure suspects) and the control group (6 primary angle-closure suspects) did not differ significantly (p = 0.560). Anterior-chamber depth (ACD) was shallower in RVO eyes (2.72 ± 0.31 mm) than in the contralateral non-RVO eyes (2.76 ± 0.31 mm; p = 0.014). CONCLUSIONS: This prospective, blinded, matched case-control study did not evidence any significant differences in clinical and AS-OCT-derived structural measures between RVO and control eyes. However, RVO eyes, compared with their contralateral non-RVO eyes, had a slightly shallower ACD. These findings collectively suggest that an association between primary angle-closure mechanisms and RVO is unlikely. However, the shallower ACD in RVO eyes could potentially put them at higher risk for intermittent or permanent pupillary block.

Dissecting Medical Litigation: An Analysis of Canadian Legal Cases in Plastic Surgery.

Introduction: Plastic surgeons are more likely to face medical litigation, compared to other specialists. Although this has been previously studied in other countries, there is a paucity of data regarding legal medical cases within Canada. The goal of this study was to compile and analyze all medical litigations in plastic surgery in Canada and identify themes associated them. Methods: A systematic search of the 2 largest Canadian online legal databases, LexisNexis Canada and WestLawNext Canada, was conducted to retrieve all legal medical cases against plastic surgeons in Canadian courts. Quantitative and qualitative analyses were performed to dissect the characteristics of plastic surgery litigation in Canada. Results: A total of 105 legal cases were included in this analysis, including 81 lawsuits and 24 appeals. The preponderance of cases was related to breast surgeries (47.0%), followed by head and neck surgeries (18.1%), with 76.5% being related to cosmetic surgery; 64.2% were ruled in favour of the surgeon. The lack of preoperative informed consent was highly associated with a final ruling in favour of the patient (P < .0001). The average monetary value of damages awarded was $61 076. There was no significant difference in monetary value between cosmetic and reconstructive cases. Conclusion: The majority of medical litigation in plastic surgery in Canada is associated with cosmetic surgeries, most commonly of the breast. Lack of informed consent is associated with judicial rulings in favour of patients. By understanding the themes underlying these legal cases, we hope to highlight the main issues that lead to litigation in plastic surgery.

Introduction: Les chirurgiens plastiques sont plus susceptibles de faire face à un contentieux médical que d'autres spécialistes. Si cela a déjà été étudié dans d'autres pays, il y a peu de données sur les affaires judiciaires de nature médicale au Canada. L'objectif de cette étude était de compiler et analyser les contentieux médicaux en chirurgie plastique au Canada et d'identifier les thèmes qui leur sont associés. Méthodes: Une recherche systématique a été menée dans les deux grandes bases de données juridiques canadiennes sur internet, LexisNexis® Canada et WestLawNext® Canada, pour en tirer toutes les affaires judiciaires contre des chirurgiens plastiques passées devant les tribunaux canadiens. Des analyses quantitatives et qualitatives ont été réalisées pour disséquer les caractéristiques des contentieux en chirurgie plastique au Canada. Résultats: Un total de 105 cas judiciaires ont été inclus dans cette analyse, dont 81 poursuites en justice et 24 appels. Une prépondérance de cas avait trait à la chirurgie mammaire (47,0 %), suivie par la chirurgie de la tête et du cou (18,1 %), et 76,5 % étant liés à chirurgie cosmétique. Dans 64,2 % des cas, la décision a été en faveur du chirurgien. L'absence de consentement éclairé préopératoire a été fortement associée à des jugements définitifs en faveur des patients (P < 0,0001). La valeur monétaire moyenne des dommages et intérêts accordés était de 61 076 $. Il n'y a pas eu de différence significative de valeur monétaire entre les cas cosmétiques et la chirurgie reconstructrice. Conclusion: La majorité des cas de contentieux médicaux en chirurgie plastique au Canada est associée à la chirurgie cosmétique, le plus souvent la chirurgie mammaire. L'absence de consentement éclairé est associée aux décisions judiciaires favorables aux patients. En comprenant les thèmes sous-tendant ces affaires judiciaires, nous espérons souligner les principaux problèmes qui débouchent sur un contentieux en chirurgie plastique.

Study of the Apoptotic Impacts of Hazelnut Oil on the Colorectal Cancer Cell Line.

Background: Hazelnut oil has a unique structure with a high oleic acid content, tocopherol, tocotrienols, and other bioactive compounds, such as phytosterols. These biochemical compounds have been widely studied because of their potential health properties. Understanding the process of apoptosis is the basis of new therapies contributing to cancer cells' death. Recently, the potential role of the evolutionary-reserved bcl-2 protein family in tumor progression and prognosis of some malignancies has been addressed in several studies. The present study is aimed at evaluating the effect of apoptotic properties of hazelnut oil on colorectal cancer cells through the major members of this family (bax and bcl-2). Materials and Methods: MTT assay, apoptotic cell staining (using Annexin V and propidium iodide), flow cytometry, and real-time PCR were used to evaluate the toxicity, percentage of apoptotic cells, and bax and bcl-2 genes' expression after exposing HT29 cells to hazelnut oil. Results: After hazelnut treatment, significant decreases in cell viability, and the gene expression of bax and bcl-2 were observed compared to the control group (P < 0.05). In addition, the total percentage of apoptotic cells after hazelnut oil treatment showed a significant increase in comparison with the negative control group (P < 0.05). Conclusion: Hazelnut oil appears to cause the death of cancerous cells through an apoptotic mechanism.

Cell Surface Vimentin Detection in Cancer Cells by Peptide-Based Monoclonal Antibody.

Background: Vimentin is a prominent Intermediate Filaments (IFs) protein expressed in different mesenchymal origin cell types. Besides a wide range of cellular function roles associated with vimentin expression, its dysregulation and cell surface expression in the induction of malignancy properties have been reported extensively, making it a promising cancer-specific target. Therefore, this study aimed to generate and characterize anti-vimentin monoclonal antibodies. Methods: A 14-mer synthetic peptide from vimentin was conjugated to Keyhole Limpet Hemocyanin (KLH) and used for immunization of Blab/C mice and monoclonal production by conventional hybridoma technology. The monoclonal antibody was purified using affinity chromatography of supernatants from the selected hybridoma cells. ELISA, Immunoprecipitation-Western blotting (IP-WB), Immunocytochemistry (ICC), and flow cytometry were employed to characterize the produced monoclonal antibody in terms of interaction with vimentin immunizing peptide as well as vimentin protein. Results: Amid the several obtained producing anti-vimentin antibody hybridomas, the 7C11-D9 clone (IgG1 isotype with kappa light chain) showed higher reactivity with the immunizing peptide, and led to its selection for purification and characterization. The purified antibody could detect vimentin protein in IP-WB, ICC and flow cytometry of the normal and cancerous cells with different origin. No vimentin expression was found in normal healthy Peripheral Blood Mononuclear Cell (PBMC). Conclusion: Taken together, 7C11-D9 anti-vimentin monoclonal antibody might be used as immune diagnostic or immune therapeutic tool where detection or targeting of vimentin in a wide range of organisms is required.

Combined Application of Human Amniotic Membrane Mesenchymal Stem Cells and a Modified PGS-co-PCL Film in an Experimental Model of Myocardial Ischemia-Reperfusion Injury.

According to the World Health Organization (WHO), about 3.9 million people die annually of ischemic heart disease (IHD). Several clinical trials have shown that stem cell therapy is a promising therapeutic approach to IHD. Human amniotic membrane mesenchymal stem cells (hAMSCs) positively affect the repair of myocardial ischemia-reperfusion (MI/R) injury by stimulating endogenous repair mechanisms. The differentiated hAMSCs with and without modified PGS-co-PCL film were applied in the myocardium. MI/R injury was induced by ligating the left anterior descending artery in 48 male Wistar rats. The rats were divided into four groups, (n = 12) animals: heart failure (HF) as the control group, HF + MSCs, HF + MSCs + film, and HF + film. Echocardiography was performed 2 and 4 weeks after MI/R injury moreover the expression of the VEGF protein was assessed in the rat heart tissue via immunohistochemistry. In vitro, our result shows fantastic cell survival when seeded on film. In vivo, the left ventricle ejection fraction (LEVD), fractional shortening (FS), end-diastolic (EDV), and stroke volume (SV) have been increased and systolic volumes decreased in all treatment groups in comparison with control. Although combination therapy has a more positive effect on hemodynamic parameters, there is no significant difference between HF + MSCs + film with other treatment groups. Also, In the IHC assay, expression of the VEGF protein significantly increased in all intervention groups. The implantation of MSCs and the modified film significantly enhanced the cardiac functional outcome; in this regard, enhancement in cell survival and VEGF expression are involved as underlying mechanisms in which cardiac film and MSCs exert a beneficial effect.

Influence of trans-ferulic acid on the quality of ram semen upon cold preservation.

BACKGROUND: Due to lower antioxidant capacity and higher amounts of polyunsaturated fatty acids, ram spermatozoa are very susceptible during cooling process. OBJECTIVES: The objective was to examine the effect of the trans-ferulic acid (t-FA) on the ram semen during liquid preservation. METHODS: Semen samples were collected from the Qezel rams, pooled, and extended with the Tris-based diluent. Pooled samples enriched with different amounts of the t-FA (0, 2.5, 5, 10, and 25 mM) and preserved at 4°C for 72 h. Spermatozoa's kinematics, membrane functionality, and viability were assessed by CASA system, hypoosmotic swelling test, and eosin-nigrosin staining, respectively. Moreover, biochemical parameters were measured at 0, 24, 48, and 72 h. RESULTS: Results showed that 5 and 10 mM t-FA improved forward progressive motility (FPM) and curvilinear velocity compared to the other groups at 72 h (p < 0.05). Samples treated with 25 mM t-FA showed the lowest total motility, FPM, and viability at 24, 48, and 72 h of storage (p < 0.05). Higher total antioxidant activity levels were observed in the 10 mM t-FA-treated group compared to the negative control at 72 h (p < 0.05). Treatment with 25 mM t-FA increased malondialdehyde amounts and decreased superoxide dismutase activity compared to other groups at the final time assessment (p < 0.05). Nitrate-nitrite and lipid hydroperoxides values were not affected by treatment. CONCLUSIONS: The current study indicates the positive and negative influences of different concentrations of t-FA on the ram semen upon cold storage.

A novel co-culture assay to evaluate the effects of sympathetic innervation on vascular smooth muscle differentiation.

Dedifferentiation of vascular smooth muscle cells (VSMCs) from a functional phenotype to an inverse synthetic phenotype is a symptom of cardiovascular disorders, such as atherosclerosis and hypertension. The sympathetic nervous system (SNS) is an essential regulator of the differentiation of vascular smooth muscle cells (VSMCs). In addition, numerous studies suggest that SNS also stimulates VSMCs to retain their contractile phenotype. However, the molecular mechanisms for this stimulation have not been thoroughly studied. In this study, we used a novel in vitro co-culture method to evaluate the effective cellular interactions and stimulatory effects of sympathetic neurons on the differentiation of VSMCs. We co-cultured rat neural-like pheochromocytoma cells (PC12) and rat aortic VSMCs with this method. Expression of VSMCs contractile genes, including smooth muscle actin (acta2), myosin heavy chain (myh11), elastin (eln), and smoothelin (smtn), were determined by quantitative real-time-PCR analysis as an indicator of VSMCs differentiation. Fold changes for specific contractile genes in VSMCs grown in vitro for seven days in the presence (innervated) and absence (non-innervated) of sympathetic neurons were 3.5 for acta2, 6.5 for myh11, 4.19 for eln, and 4 for smtn (normalized to Tata Binding Protein (TBP)). As a result, these data suggest that sympathetic innervation promotes VSMCs' contractile gene expression and also maintains VSMCs' functional phenotype.

A Novel Nanocomposite Scaffold Based on Polyurethane (PU) Containing Cobalt Nanoparticles (CoNPs) for Bone Tissue Engineering Applications.

BACKGROUND: Bone tissue engineering, as a relatively new approach, has focused on combining biodegradable scaffolds, cells, and biologically active molecules for the recovery of different damaged tissues, such as bone defects. Polyurethane (PU), as a synthetic polymer, benefits from a porous structure which impersonates bone's natural environment. However, PU lacks osteoinduction activities. Cobalt nanoparticles (CoNPs) stimulate angiogenesis and biomineralization, which greatly favors osteogenesis. METHODS: Here, we designed a novel scaffold based on PU and combined it with CoNPs for bone regeneration applications. The composition and structure of PU-CoNPs nanocomposite were characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and differential scanning calorimetry (DSC). MTT and AO data showed biocompatibility and enhanced viability and proliferation of fibroblasts on PU-CoNPs scaffold. Ascorbic acid-2-phosphate, ß-glycerophosphate, and dexamethasone-induced osteogenesis for 14 days. RESULTS: The alkaline phosphatase test asserts the increased mineralization of hADSCs cultured on PUCoNPs compared to pure PU scaffold. Further, the results disclosed an elevated osteogenic differentiation at the level of genes and proteins using immunocytochemical analysis (ICC) and quantitative real-time PCR (qPCR). CONCLUSION: These findings provide an evidence that PU-CoNPs nanocomposite might be a promising candidate for bone repair applications.

Attenuation Correction for Dedicated Cardiac SPECT Imaging Without Using Transmission Data.

Objectives: Attenuation correction (AC) using transmission scanning-like computed tomography (CT) is the standard method to increase the accuracy of cardiac single-photon emission computed tomography (SPECT) images. Recently developed dedicated cardiac SPECT do not support CT, and thus, scans on these systems are vulnerable to attenuation artifacts. This study presented a new method for generating an attenuation map directly from emission data by segmentation of precisely non-rigid registration extended cardiac-torso (XCAT)-digital phantom with cardiac SPECT images. Methods: In-house developed non-rigid registration algorithm automatically aligns the XCAT- phantom with cardiac SPECT image to precisely segment the contour of organs. Pre-defined attenuation coefficients for given photon energies were assigned to generate attenuation maps. The CT-based attenuation maps were used for validation with which cardiac SPECT/CT data of 38 patients were included. Segmental myocardial counts of a 17-segment model from these databases were compared based on the basis of the paired t-test. Results: The mean, and standard deviation of the mean square error and structural similarity index measure of the female stress phase between the proposed attenuation maps and the CT attenuation maps were 6.99±1.23% and 92±2.0%, of the male stress were 6.87±3.8% and 96±1.0%. Proposed attenuation correction and computed tomography based attenuation correction average myocardial perfusion count was significantly higher than that in non-AC in the mid-inferior, mid-lateral, basal-inferior, and lateral regions (p<0.001). Conclusion: The proposed attenuation maps showed good agreement with the CT-based attenuation map. Therefore, it is feasible to enable AC for a dedicated cardiac SPECT or SPECT standalone scanners.