Comparison of T Lymphocyte Subsets and Natural Killer Lymphocytes in Moderate Versus Severe COVID-19 Patients.

Severe respiratory involvement that follows a process of immune dysregulation and intense cytokine production remains to be the most dreaded complication of Coronavirus Disease-2019 (COVID-19) infection. The aim of this study was to analyze T lymphocyte subsets and natural killer (NK) lymphocytes in moderate and severe cases of COVID-19 infection and assess their significance in disease severity and prognosis. Twenty moderate cases and 20 severe cases of COVID-19 were studied and compared regarding blood picture, biochemical markers, T lymphocyte population subsets, and NK lymphocytes, which were determined by flow cytometric analysis. On analyzing the flow cytometric data of T lymphocyte cells and their subsets and NK cells in two groups of COVID-19 infection (one group moderate and the other severe cases), some immature NK lymphocyte relative and absolute counts were higher in the severe patients with worse outcome and death, while some mature NK lymphocyte relative and absolute counts were depressed in both groups. Also, interleukin (IL)-6 was significantly higher in severe cases when compared to moderate cases, and there was a positive significant correlation between immature NK lymphocyte relative and absolute counts and IL-6. There was no statistically significant difference between T lymphocyte subsets (T helper and T cytotoxic) with disease severity or outcome. Some immature NK lymphocyte subsets contribute to the widespread inflammatory response that complicates severe cases of COVID-19; therapeutic approaches directed to enhancing NK maturation or drugs that block NK cell inhibitory receptors have a potential role in controlling COVID-19 induced cytokine storm.

Therapeutic efficacy of dendritic cell injection in advanced hepatocellular carcinoma: the role of natural killer and T lymphocytes.

Aim of the study: Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related fatalities worldwide. The burden of HCC incidence in Egypt has doubled in the last 10 years. The primary aim of this research was to assess the safety and efficacy of autologous dendritic cells (DCs) generated from peripheral blood. Material and methods: This trial was carried out at the Sohag Center of Cardiac and Digestive System. Patients with HCC were grouped into two groups (control group and DC injection group). The study group received intradermal autologous DCs twice weekly for three weeks, with a total of six vaccinations of 0.7 IU, whereas the control group received conservative treatment. Results: The study group showed statistically significant clinical improvement in the Child-Pugh score and overall survival. Laboratory evaluation revealed a significant reduction of α-fetoprotein, from 232 ng/dl at baseline to 193 ng/dl after 3 months to 153 ng/dl after 6 months, in the injection group, as compared with the control group, which increased from 228 ng/dl at baseline to 269 ng/dl at 3 months to 305 ng/dl at 6 months. Also, liver function improved significantly at both 3 and 6 months in the injected group compared with the control group. Regarding lymphocyte subsets, T-cytotoxic lymphocytes (CD8+) and natural killer cells (CD56+ve) increased significantly in the injection group. Conclusions: DC injection may be effective treatment of patients with advanced HCC to improve quality of life.

Video game disorder and mental wellbeing among university students: a cross-sectional study.

Introduction: video games are a popular adult pastime but have a potentially pervasive negative influence on gamers. The aim: was to determine the prevalence of video game disorder (VGD), its associated predictors, and its impact on the mental health of university students. Methods: a cross-sectional study was performed with a convenience sample of 2,364 undergraduate students. Sociodemographic criteria, the Internet Gaming Disorder-20 (IGD-20) questionnaire, and the Mental Health-5 (MH-5) questionnaire were used to collect data. The weights and heights of the students were self-reported. The average number of hours spent playing video games per week, the average number of hours of sleep per day, the favorite type of game played, and the main causes for playing were also included. Results: the prevalence of VGD among participants was 18.9%. The main predictors of VGD were being male, residing in an urban area, playing more hours per week, sleeping fewer hours per day, and having a higher body mass index, while having a low socioeconomic status was a protective factor. Mental health had a strong negative correlation with VGD. The types of games most frequently played by video game addicts were violent and action games. However, the most frequent reasons cited for playing were to improve one´s avatar, relaxation, and amusement. Conclusion: playing video games in moderation, adequate sleeping, and engaging in outdoor physical activities enhances mental well-being and physical functioning. Thus, it is critical to promote and encourage balanced, effective, stable approaches to video gaming among university students to maintain their mental well-being.

The relation of CD4+CD25+Foxp3+ regulatory T cells concentration with disease activity and damage index in systemic lupus erythematosus.

INTRODUCTION: Regulatory T cells (Treg) deficits, both quantitative and qualitative, are known to be possible triggers for the development of autoimmune disorders by causing T and B cells dysfunction. The contribution of Treg deficiency in the etiology of systemic lupus erythematosus (SLE) is still being debated. The aim of the present study is to evaluate the percentage of circulating CD4+CD25+Foxp3+ Treg cells in a cohort of Egyptian SLE patients and to correlate this value with the activity and damage index of these patients. METHODS: 50 female patients with SLE together with an equal number of age- and sex-matched healthy controls were enrolled in the study. Flow cytometric determination of peripheral Treg cells was carried out for all participants by detecting the percentage of CD4+CD25+Foxp3+ cells to compare cases with the control group. Disease activity was assessed using the Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K), while disease damage was assessed using the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SLICC/ACR DI). Both indices were correlated with the percentage of CD4+CD25+Foxp3 T regulatory cells. RESULTS: CD4+CD25+Foxp3+ Treg cells percentage was significantly decreased in patients with SLE as compared to healthy controls. On correlating CD4+CD25+Foxp3+ Treg percentage with SLEDAI-2K, a significantly negative correlation was found. Also, there was a negative significant correlation between CD4+CD25+Foxp3+ Treg cells percentage and SLICC/ACR DI. On correlating SLEDAI-2K with damage index (SLICC/ACR DI), we found highly significant positive correlation. CONCLUSION: Our study showed impaired production of CD4+CD25+Foxp3+ Tregs in SLE patients, which can play a reciprocal role with some cytokines to affect the activity of the disease and organ damage. CD4+CD25+Foxp3+ Tregs cells should be the target to determine the clinical effectiveness of new therapy to modulate Tregs besides the traditional treatments.

Changes in peripheral blood cellular morphology as diagnostic markers for COVID-19 infection.

INTRODUCTION: Real-time reverse-transcriptase polymerase chain reaction (RT-PCR) assays were established to detect severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2). However, due to the high rate of false negative results, additional tests as computed tomography (CT) scans of the chest and blood chemistry are required to properly diagnose COVID-19 infection. Abnormal morphological changes of peripheral blood cells as granulocytic dysmorphism and abnormal reactive lymphocytes have been described in some cases. The aim of the present study was to investigate the morphological changes affecting all peripheral blood cells of COVID-19 patients, in order to find any specific abnormalities that could help in the early diagnosis and/or prognosis. METHODS: Peripheral blood smears of 113 COVID-19 patients and 50 non-COVID-19 controls were examined for morphological changes in the period between October 2020 and January 2021 (second wave). We set a score value in which every morphological abnormality was given one point in each examined blood smear. Score, neurophil/lymphocyte (N/L) ratio, and blood chemistry were compared to the severity and outcome of the disease. RESULTS: Significant morphological changes were found when compared to control blood smears. Various abnormalities as pyknotic cells, broken cells, pseudo Pelger-Huët, abnormal lymphocytes, abnormal monocytes, and leukoerythroblastic reaction were found. Cases with higher scores had unfavorable outcomes (p = .005). High interleukin-6 (IL-6) levels were correlated to pyknotic cells (p = .003). CONCLUSION: The blood picture of COVID-19 patients revealed various morphological changes that are not detected with the same frequency and variability in other viral infections. The prominent morphological changes can be predictive of an undesirable outcome of the disease.

Therapeutic potential of mesenchymal stem cells for scleroderma induced in mouse model.

OBJECTIVE: To examine the potential therapeutic effect of mesenchymal stem cells (MSCs) for experimental scleroderma. MATERIALS AND METHODS: Fifty-four mice six-week-old (30-35 g) were studied. Hypochlorous acid (HOCl) induced scleroderma was considered. Mice were divided into 3 groups: (I) Control: Six mice did not receive any treatment and were sacrificed at the end of the experiment; (II) HOCl mice (induced scleroderma as a positive control): (III) MSCs-treated HOCl mice: Thirty six HOCl-induced mice were injected with MSCs (7.5 × 105) intravenous every week for 3 weeks. Skin pieces were taken from the backs of mice and lung tissue pieces. a smooth muscle actin (α-SMA) and transforming growth factor-ß (TGF-ß1) were analysed or fixed in 10 % formalin for skin and lung tissue histopathological analysis. Plasma nitric oxide (NO) was also assayed. RESULTS: There was a significant rise in the NO level and of the cutaneous and lung tissue α-SMA and TGF-ß1 in untreated scleroderma-induced mice. The values significantly normalized after MSC therapy over the 7 weeks duration of the study. The altered histopathology of the skin and lung tissues in the scleroderma-induced mice showed a remarkable tendency to normalization of the skin and lung parenchyma and vasculature. CONCLUSION: There was a significant rise in the level of NO and skin and lung tissue α-SMA and TGF-ß1 in untreated scleroderma-induced mice and values were significantly normalized after MSC therapy over the 7 weeks duration of the study. Altered histopathology of the skin and lung appeared nearly normal after MSC therapy.

Implanted subcutaneous versus intraperitoneal bioscaffold seeded with hepatocyte-like cells: functional evaluation.

BACKGROUND AND OBJECTIVES: The X-linked bleeding disorder, hemophilia A, is caused by defective production of factor VIII (FVIII). Hemophilic patients require regular FVIII infusions. Recombinant factor replacement poses the safest line of therapy. However, its main drawbacks are high expenses and the higher liability for formation of inhibitors. Recent studies confirmed the ability of bone marrow-derived stem cells to secrete FVIII. This study aims to generate bioscaffold from decellularized liver and subsequently seed it with trans-differentiated human stem cells into hepatic-like cells. This scaffold can then be implanted intraperitoneally or subcutaneously to provide FVIII. METHODS: After generation of the bioscaffold, seeding of discoid scaffolds with trans-differentiated human hepatocyte-like cells was performed. Then, the generated organoid was implanted into peritoneal cavity or subcutaneous tissue of experimental rats. RESULTS: Serum human FVIII was significantly increased in rats subjected to subcutaneous implantation compared intraperitoneal implantation. Immunostaining for detecting Cytokeratin 19 and human anti-globulin confirmed the presence of mature human hepatocytes that were significantly increased in subcutaneous implanted scaffold compared to the intraperitoneal one. CONCLUSION: Implantation of decellularized bioscaffold seeded with trans-differentiated stem cells in rats was successful to establish production of FVIII. Subcutaneous implantation showed higher FVIII levels than intraperitoneal implantation.

Effects of remote virtual work environment during COVID-19 pandemic on technostress among Menoufia University Staff, Egypt: a cross-sectional study.

Technostress during the COVID-19 pandemic has become more prevalent as a result of the global preventive measures applied to limit the spread of infection. These measures included remote working from home in both public and private organizations. The objective of this study is to study technostress and challenges of remote virtual work environment among university staff members at Menoufia University, Egypt. A cross-sectional study was conducted over Menoufia University academic staff members in Egypt. The participants were chosen from both practical and theoretical colleges in Menoufia University using a multistage random sample. Tarfadar technostress questionnaire was used. Cortisol blood level was measured for all participants. This study included 142 participants. The mean age of the group was 36.32±6.41 years. 52.1 percent worked in practical colleges, and 60.6% were lecturers or higher. Their mean cortisol level was 15.61±7.07mcg/dl. Participants who were females, reside in rural areas, held a lecturer or higher position, had poor work-environment WiFi, and lacked technical training had significantly higher levels of technostress subscales. Most of the technostress subscales were significantly correlated with age and blood cortisol levels. The predictors of work overload in multivariate regression were female gender and a work environment with poor WiFi. Female gender, theoretical colleges, being lecturer or higher, and poor WiFi were the predictors for invasion. Among university staff members, technostress was found to be evident. High levels of technostress were significantly influenced by age, higher professions, female gender, and a bad workplace environment.

CRISPR-mediated gene modification of hematopoietic stem cells with beta-thalassemia IVS-1-110 mutation.

BACKGROUND: ß-Thalassemias represent a group of genetic disorders caused by human hemoglobin beta (HBB) gene mutations. The radical curative approach is to correct the mutations causing the disease. CRISPR-CAS9 is a novel gene-editing technology that can be used auspiciously for the treatment of these disorders. The study aimed to investigate the utility of CRISPR-CAS9 for gene modification of hematopoietic stem cells in ß-thalassemia with IVS-1-110 mutation. METHODS AND RESULTS: We successfully isolated CD34+ cells from peripheral blood of ß-thalassemia patients with IVS-1-110 mutation. The cells were transfected with Cas9 endonuclease together with guide RNA to create double-strand breaks and knock out the mutation. The mutation-corrected CD34+ cells were subjected to erythroid differentiation by culturing in complete media containing erythropoietin. CONCLUSION: CRISPR/Cas-9 is an effective tool for gene therapy that will broaden the spectrum of therapy and potentially improve the outcomes of ß-thalassemia.

Functional beta-cells derived from umbilical cord blood mesenchymal stem cells for curing rats with streptozotocin-induced diabetes mellitus.

INTRODUCTION: This study aimed to investigate the therapeutic response to injected human umbilical cord blood mesenchymal stem cells (UCBMSCs) among albino rats with streptozotocin (STZ)-induced diabetes mellitus. METHODS: Control group (GI; n = 25) rats were fed with standard rat diet. Rats with STZ-induced diabetes mellitus without (GII; n = 25) and with (GIII; n = 25) differentiated human UCBMSCs implantation were the test groups. Rats were sacrificed in Week 11 following implantation. Liver biopsies were sectioned and stained in order to highlight both the presence and function of impregnated cells in the liver tissue. RESULTS: Haematoxylin and eosin-stained sections in GI and GII rats showed normal liver architecture while GIII rats showed presence of cell clusters inside the liver tissue and around the central veins. Cell clusters with blue cytoplasm were present in sections in GIII rats but absent in GI and GII rats, indicating the presence of injected differentiated human UCBMSCs. The anti-human insulin immunostaining of GIII rats showed clusters of cells within the liver parenchyma and around central veins, indicating that these cells were active and secreting insulin. CONCLUSION: UCBMSCs are proficient in differentiating into insulin-producing cells in vivo under specific conditions and, when transplanted into the liver of albino rats with STZ-induced diabetes mellitus, were able to secrete insulin and partially control the status of diabetes mellitus in rats.

Decellularized liver bioscaffold: a histological and immunohistochemical comparison between normal, fibrotic and hepatocellular carcinoma.

AIM OF THE STUDY: Increasing demand for liver transplantation represents an important health burden. Decellularized liver bioscaffold can be a suitable alternative for whole organ transplantation. However, various pathologies can affect the structure of decellularized scaffolds.This work discusses differences between hepatic fibrosis (HF), hepatocellular carcinoma (HCC) and normal decellularized liver bioscaffolds. MATERIAL AND METHODS: Murine models of HF and HCC were created, livers from normal, HF and HCC were decellularized, and evaluation of decellularization was done using morphological, histological and DNA analysis examination. Also, immunohistochemical staining using collagen, laminin, fibronectin and alphafetoprotein was done. Deposition area and intensity of the used immunohistochemical staining in liver capsules and the staining deposition thickness in the blood vessels and hepatic capsule walls were measured for comparison between the three models. RESULTS: Normal, HF and HCC livers were decellularized efficiently as confirmed by histological and DNA estimation. HCC decellularized samples showed significantly higher collagen, fibronectin and laminin deposition in both capsule and blood vessels, followed by HF decellularized samples, which also showed the highest thickness of laminin deposition in both capsule and blood vessels, then the normal model, which recorded the lowest value. Alphafetoprotein positive cells were absent in normal and HF, with rare cells in HCC. CONCLUSIONS: Even pathologic livers, HF and HCC, can be efficiently decellularized, showing normal morphology and architecture. However, HCC and HF showed significantly higher deposition of extracellular matrix proteins: collagen, fibronectin and laminin. The impact of these differences on physiological and immunological functions of the bioscaffold requires recellularization experiments.

Risk Factors Associated with Needlestick Injuries among Health Care Workers in Menoufia Governorate, Egypt.

BACKGROUND: Health care workers are at high risk of job-related blood-borne diseases due to needlestick injuries (NSIs). OBJECTIVE: To assess the risk factors associated with NSIs among health care workers in Menoufia governorate, Egypt. METHODS: This cross-sectional study was conducted on 2260 health care workers of 4 randomly chosen hospitals in Menoufia governorate. Using a predesigned data collection sheet, all staff members were asked about the occurrence of NSIs in the previous 3 months. The response rate was 95.3%. Logistic regression analysis was used to assess the factors associated with NSIs. RESULTS: The risk of NSIs significantly increased with duration of work <15 years (OR 2.19, 95% CI 1.81 to 2.66), being female (OR 1.89, 95% CI 1.56 to 2.29), working as a paramedic (OR 1.49, 95% CI 1.03 to 2.25), working in surgical ward (OR 4.11, 95% CI 1.71 to 9.88), having more than 2 night shifts/month (OR 1.75, 95% CI 1.28 to 2.39), absence of educational sessions (OR 1.99, 95% CI 1.45 to 2.73), absence of hospital policies for NSIs (OR 2.23, 95% CI 1.99 to 2.49), absence of universal precautions (OR 1.66, 95% CI 1.10 to 2.50), recapping the needle after use (OR 2.63, 95% CI 2.12 to 3.26), recapping the needle with two hands (OR 3.08, 95% CI 2.04 to 4.65), not using protective clothes (OR 1.39, 95% CI 1.04 to 1.85), and increased working hours---8-12 hours (OR 2.14, 95% CI 1.34 to 3.44) and >12 hours (OR 2.28, 95% CI 1.17 to 4.44). CONCLUSION: The risk of NSIs is still high among health care workers that underlines the importance of comprehensive educational sessions to decrease the risk of job-related blood-borne diseases.

Role of cord blood and bone marrow mesenchymal stem cells in recent deep burn: a case-control prospective study.

RATIONALE: Burn injuries represent one of the major worldwide public health problems causing more severe physiological stress than other traumas. Effective treatment of burn injuries is mandatory to prevent the numerous life-threatening complications and possible disabilities. Stem cells, a population of multipotent cells retaining the properties of self-renewal and differentiation, are the main player in tissue regeneration after major trauma. Thus, they are thought to play a key role in wound healing inducing efficient and physiological skin regeneration. Stem cell-based regeneration is quickly gaining scientific grounds. OBJECTIVE: This study was designed as a comparative prospective study to evaluate and compare the regenerative effect of bone marrow derived mesenchymal stem cells (BM-MSCs) and umbilical cord blood derived mesenchymal stem cells (UC-MSCs) compared to conventional early excision and graft (EE&G) in recent thermal full thickness burned patients. SUBJECT & METHODS: Recruited burned patients were randomly divided into three groups (20 patients on each group) having recent thermal full thickness percentage ranging from 10% to 25% total body surface area (TBSA). After receiving allocated treatment, they were assessed as regards: rate of burn healing, presence of post-burn complications both early (such as loss of graft and infections) and late (as hypertrophic scars, keloid, hypo- or hyperpigmentation or contracture of the wound), hospitalization time and cost. RESULTS: This study showed significantly improved rate of healing in both BM-MSC and UC-MSC groups as compared to EE&G group with no significant difference between bone marrow and umbilical cord groups. Comparing the incidence of early complications, partial and total loss of graft occurred in 50% patients in (EE&G) group, while infection complication appeared in 25% of patients of (BM-MSCs) group and in 70% of patients in (UC-MSCS) group. The late complications (hypertrophic scars) were observed in 40% of (EE&G) patients group, in 15% of (BM-MSCs) treated patients group and 20% of (UC-MSCS) patients group. Contractured scars were present in 15% in (EE&G) group, 10% in (BM-MSCs) group, 10% in (UC-MSCS) group. Hypopigmentation occurred in 20% of patients in (EE&G) group, 20% in (BM-MSCs) group and 10% in (UC-MSCS) group. Hyperpigmentation was present in 20% of patient in (EE&G) group, 30% in (UC-MSCS) group but no hyperpigmentation occurred in (BM-MSCs) group. There was no late complication in 5% of patient in (EE&G) group, 55% in (BM-MSCs) group and 30% in (UC-MSCS) group. The results of this study revealed that the hospitalization period was significantly reduced in both (BM-MSCs) group and (UC-MSCS) group as compared to (EE&G) group. CONCLUSION: this study proves that mesenchymal stem cells, both from bone marrow and cord blood origin, can effectively improve healing of burn injuries.

Chimerism in pediatric hematopoietic stem cell transplantation and its correlation with the clinical outcome.

Hematopoietic stem cell transplantation (HSCT) is the only hope to cure many inherited and acquired hematological disorders in children. Monitoring of chimerism helps to predict the post-transplantation events, with the intention to enhance the long-term disease free survival (DFS). The study aimed to investigate the importance of early chimerism detection to predict the clinical outcome following HSCT. The study included nine recipients (six ß-thalassemia and three severe aplastic anemia patients) and their 10/10 HLA identical sibling donors. Chimerism detection was performed by analysis of short tandem repeat (STR) polymerase chain reaction (PCR) for detection and quantification of the relative amounts of donor and recipient cells present on day +28. Peripheral blood (PB) was the main stem cell source for HSC transplantation. Disease free survival (DFS) was 71.4% while overall survival was 85.7% for PBSC transplants at the median follow up period of 4years. The early detection of chimerism by PCR-STR analysis for children with ß-thalassemia and aplastic anemia correlated with the outcome of HSCT in 8 (88.8%) patients. Complete chimerism was associated with disease-free survival while mixed chimerism and autologous patterns were associated with poor prognosis. In conclusion, early chimerism testing is clinically important in prediction of outcome after allogeneic HSC transplantation.

Targeting Future Customers: An Introductory Biobanking Course for Undergraduate Students of Life Sciences.

INTRODUCTION: Biobanking is a relatively new concept in the Arab region. Targeting different stakeholders to introduce the concept of biobanking and develop an acceptance of it among them is important for the growth of biobanking in the region. Undergraduate students of life sciences represent an important segment of stakeholders, since they constitute potential future biobank customers. Limited funding, lack of awareness of the existence of the term "biobanking" itself among these students, and questions regarding best marketing strategies presented challenges to planning for the most effective message delivery to this target group. METHODS: A specific course was designed for undergraduate students of life sciences, which was conducted at the Faculty of Medicine, Cairo University, Egypt. The course was conducted twice in 2016 and included lectures covering biobanking, quality, ethics, information technology, and translational research. Facebook and word-of-mouth were used for marketing and advertising. RESULTS: A total number of 125 participants attended both courses cumulatively. Facebook appeared to have been an effective marketing outlet, especially when paid advertisements were used. Evaluation of knowledge, measured using a pretest and posttest, demonstrated some improvement in knowledge of participants. Evaluation forms filled after the course showed positive attitude toward content and message delivery by a majority of participants. Facebook was also used as an evaluation method through analysis of engagement with posts created after course completion. CONCLUSIONS: Biobanking education can be carried out effectively with limited resources. Understanding the needs of the target group and using appropriate methods of communication are essential prerequisites to a well-tailored curriculum and effective message delivery. Using Facebook appears to be an effective and affordable method of communication and advertising. Targeting undergraduate students of life sciences interested in research is a good investment and can be very effective in increasing awareness about biobanking inside the research community.

Enhancement of neural regeneration after spinal cord injury using muscle graft in experimental dogs.

AIMS: Spinal cord injuries (SCIs) can cause severe disability or death. The principal treatments for traumatic SCI include surgical stabilization and decompression. Using muscle as a scaffold is a new approach. The aim of this work is to evaluate the clinical efficacy of muscle graft as a scaffold for the growing axons organizing their growth, preventing gliosis in the damaged area and enhancing neural recovery in canine model of traumatic spinal cord injury. METHODS: 14 dogs were divided into group I (Control group) 4 control dogs subjected to Sham operation, group II (Trauma control group) 5 dogs subjected to dorsal laminectomy with excision of 1 cm segment of the spinal cord and group III (Muscle graft group) 5 dogs subjected to dorsal laminectomy then muscle graft was taken from the longissimus thoraces and inserted into the spinal cord gap. The animals of all groups were euthanatized after 8 weeks. Olby and modified Tarlov scores were used to clinically evaluate the therapeutic effects. Spinal cord specimens were subjected to histological, morphometric and statistical studies. RESULTS: Olby and modified Tarlov scores revealed significant clinical improvement in the muscle graft group. Histological sections showed overgrowth of axons on the muscle graft and the sections started to organize as central gray matter and peripheral white matter. CD44 & CD105 stains were positive for endogenous stem cells. CONCLUSIONS: This study proved the clinical efficacy of muscle grafting as a tool for induction of neuroregeneration after traumatic spinal cord injury.

The effect of bone marrow-derived mesenchymal stem cells on chemotherapy induced ovarian failure in albino rats.

OBJECTIVES: Chemotherapy targets rapidly dividing tissues in the body. It destroys the progenitor cells in gonads resulting in premature ovarian failure. Studies have suggested that bone marrow-derived stem cells can generate oocytes in chemotherapy treated female rats after transplantation. The present study aimed to assess mechanism of homing, the action of injected BM-MSCs on ovarian function after ovarian damage. EXPERIMENTAL DESIGN: Seventy two female albino rats were randomly allocated into Control and CTX group, The Experimental protocol was lasted for 12 weeks during which serum FSH and E2 were monitored twice at the end of the 2nd week (12 rats) and 8th week (6 rats). Stem cells identification and homing were evaluated by Flowcytometry and tagging of stem cells with iron oxide particles respectively. Also, histopathological examination was done to evaluate both degeneration (6 rats at 4th week) and regeneration (6 rats at 12th week) of ovarian tissue together with assessment of the levels of TNF-α in ovarian homogenate and IGF-I as a growth factor in ovarian tissue. PRINCIPAL OBSERVATIONS: Partial improvement of E2 and FSH levels as well as ovarian architecture. Elevation of ovarian TNF- α levels and of IGF-I immunohistochemical expressions in ovarian tissues of BM-MSCs injected rats were noticed following homing of BM- MSCs in the ovarian stroma in both control and chemotherapy groups. CONCLUSION: Injected BM- MSCs can home in the stroma of the injured ovaries. IGF-I and TNF- α may have a role in the attraction of stem cells in vivo.

Treatment of dystrophic epidermolysis bullosa with bone marrow non-hematopoeitic stem cells: a randomized controlled trial.

Patients with dystrophic epidermolysis bullosa (DEB) have mutations in type VII collagen gene. Type VII collagen is synthesized by keratinocytes and fibroblasts. Based on the ability of bone marrow non-hematopoeitic stem cells (NHBMSC) to develop into fibroblasts, we decided to investigate the use of NHBMSC in the treatment of recessive DEB (RDEB). This study included fourteen patients with RDEB; the first seven of them were given cyclosporine after the infusion of NHBMSC. As cyclosporine has been used for the treatment of RDEB we decided not to use cyclosporine for the second group of seven patients. Skin biopsies from the lesions were studied by electron microscopy before and after treatment. The number of new blisters decreased significantly after treatment in both groups (p = 0.003 and 0.004 respectively) and the rate of healing of new blisters became significantly faster after treatment in both groups (p < 0.001) with no significant difference between the two groups. Electron microscopic examination revealed increased number of anchoring fibrils after treatment in both groups. No major side effects were reported during the 1-year follow-up period. Our findings highlight the efficacy as well as the safety of NHBMSC in the treatment of RDEB.

Avemar and Echinacea extracts enhance mobilization and homing of CD34(+) stem cells in rats with acute myocardial infarction.

INTRODUCTION: Activation of endogenous stem cell mobilization can contribute to myocardial regeneration after ischemic injury. This study aimed to evaluate the possible role of Avemar or Echinacea extracts in inducing mobilization and homing of CD34(+) stem cells in relation to the inflammatory and hematopoietic cytokines in rats suffering from acute myocardial infarction (AMI). METHODS: AMI was developed by two consecutive subcutaneous injections of isoprenaline (85 mg/kg). AMI rats were either post-treated or pre- and post-treated daily with oral doses of Avemar (121 mg/kg) or Echinacea (130 mg/kg). In whole blood, the number of CD34(+) cells was measured by flow cytometry and their homing to the myocardium was immunohistochemically assessed. Serum creatine kinase, vascular endothelial growth factor, interleukin-8 and granulocyte macrophage colony stimulating factor were determined on days 1, 7 and 14 after AMI. Sections of the myocardium were histopathologically assessed. RESULTS: Rats pre- and post-treated with Avemar or Echinacea exhibited substantial increases in the number of circulating CD34(+) cells, peaking on the first day after AMI to approximately 13-fold and 15-fold, respectively, with a decline in their level on day 7 followed by a significant increase on day 14 compared to their corresponding AMI levels. Only post-treatment with Echinacea caused a time-dependent increase in circulating CD34(+) cells on days 7 and 14. Such increases in circulating CD34(+) cells were accompanied by increased homing to myocardial tissue 14 days after AMI. Interestingly, pre- and post-treatment with Avemar or Echinacea substantially increased serum creatine kinase on day 1, normalized its activity on day 7 and, on continued treatment, only Echinacea markedly increased its activity on day 14 compared to the corresponding AMI values. Moreover, both treatments modified differently the elevated serum vascular endothelial growth factor and the lowered granulocyte macrophage colony stimulating factor levels of the AMI group but did not affect the level of interleukin-8. These results were supported histopathologically by reduced inflammatory reactions and enhanced neovascularization. CONCLUSION: Avemar and Echinacea extracts can effectively induce mobilization and homing of CD34(+) stem cells to the myocardial tissue and thus may help in stem cell-based regeneration of the infarcted myocardium.

Comparison of adipose tissue- and bone marrow- derived mesenchymal stem cells for alleviating doxorubicin-induced cardiac dysfunction in diabetic rats.

INTRODUCTION: Doxorubicin (DOX) is a well-known anticancer drug. However its clinical use has been limited due to cardiotoxic effects. One of the major concerns with DOX therapy is its toxicity in patients who are frail, particularly diabetics. Several studies suggest that mesenchymal stem cells (MSCs) have the potential to restore cardiac function after DOX-induced injury. However, limited data are available on the effects of MSC therapy on DOX-induced cardiac dysfunction in diabetics. Our objective was to test the efficacy of bone marrow-derived (BM-MSCs) and adipose-derived MSCs (AT-MSCs) from age-matched humans in a non-immune compromised rat model. METHODS: Diabetes mellitus was induced in rats by streptozotocin injection (STZ, 65 mg/kg b.w, i.p.). Diabetic rats were treated with DOX (doxorubicin hydrochloride, 2.5 mg/kg b.w, i.p) 3 times/wk for 2 weeks (DOX group); or with DOX+ GFP labelled BM-MSCs (2x106cells, i.v.) or with DOX + GFP labelled AT-MSCs (2x106cells, i.v.). Echocardiography and Langendorff perfusion analyses were carried out to determine the heart function. Immunostaining and western blot analysis of the heart tissue was carried out for CD31 and to assess inflammation and fibrosis. Statistical analysis was carried out using SPSS and data are expressed as mean ± SD. RESULTS: Glucose levels in the STZ treated groups were significantly greater than control group. After 4 weeks of intravenous injection, the presence of injected MSCs in the heart was confirmed through fluorescent microscopy and real time PCR for ALU transcripts. Both BM-MSCs and AT-MSCs injection prevented DOX-induced deterioration of %FS, LVDP, dp/dt max and rate pressure product. Staining for CD31 showed a significant increase in the number of capillaries in BM-MSCs and AT-MSCs treated animals in comparison to DOX treated group. Assessment of the inflammation and fibrosis revealed a marked reduction in the DOX-induced increase in immune cell infiltration, collagen deposition and αSMA in the BM-MSCs and AT-MSCs groups. CONCLUSIONS: In conclusion BM-MSCs and AT-MSCs were equally effective in mitigating DOX-induced cardiac damage by promoting angiogenesis, decreasing the infiltration of immune cells and collagen deposition.