Systemic Immune-Inflammation and Systemic Inflammation Response Indices are Predictive Markers of Mortality in Inpatients Internal Medicine Services.

Purpose: Internal medicine services serve the patient population with many chronic diseases. Therefore, it is high mortality rates compared to other departments of the hospital. Estimating the prognostic risk of hospitalized patients may be useful in mortality for patients. In this study, we evaluated the level of Systemic Immune Inflammation Index (SII) and Systemic Inflammation Response Index (SIRI) and its association with mortality in inpatients. Patients and methods: This study was performed in 2218 patients who were hospitalized between January 1st-December 31th of 2019. Patients were followed up for three years about primary endpoint as all-cause (except for unnatural deaths) mortality. Participants were divided into 4 equal groups according to their increasing levels of SII and SIRI. (Quartile 1-4) Age, gender, diabetes mellitus, hypertension, coronary artery disease, chronic kidney disease, malignancies (solid), white blood cell, neutrophil, lymphocyte, monocytes, hemoglobin, hematocrit, platelet, CRP, albumin, Systemic Inflammation Response Index (Quartile 1-4), Systemic Immune Inflammation Index (Quartile 1-4) were compared between survival and non-survival groups. Results: There were 1153 female and 1065 male participants enrolled. Compared with surviving patients, patients who died were older and had a higher prevalence of diabetes mellitus, hypertension, malignancy, chronic kidney disease and coronary artery disease (p < 0.001). There was a lower proportion of female patients among the patients who died. Compared to the survivor group, group who died exhibited a significant increase in CRP level, neutrophil, white blood cell and monocyte counts, but had a lower lymphocyte count, albumin level and hemoglobin count (P < 0.001). Results of Cox regression analysis showed that age, chronic kidney disease, malignancy, SIRI quartile 3, 4 and SII quartile 3, 4 pointed out a close relationship with mortality risk. (P < 0.001). Conclusion: The SIRI and SII have indicated the clinical importance of as novel markers for predicting mortality in inpatients.

Effects of vitrification solution supplemented with platelet-rich plasma in rat ovarian tissue cryopreservation.

Background/aim: The subject of this study was to investigate the utility of platelet-rich plasma (PRP) in the cryopreservation process to reduce cryodamage and increase tissue viability. Materials and methods: Twenty-one female Wistar rats were randomly allocated to three groups. In Group 1 (G1), rats were not subjected to vitrification (n = 7). Group 2 (G2) was the vitrification group in which PRP was added to the basic vitrification solution (n = 7). Group 3 (G3) was the vitrification group in which fetal bovine serum was added to the basic vitrification solution (n = 7). Warmed tissues were evaluated with histochemical (HC) and immunohistochemical (IHC) staining, the TUNEL method, immunofluorescence (IF) staining, and biochemical analyses. Results: The percentages of IHC staining, TUNEL method positivity, and IF staining were significantly higher in G2 compared to both G1 and G3 (P < 0.05). G2 ovaries exhibited a significant increase in both malondialdehyde and catalase values in comparison to G1 (P < 0.05). In HC staining, degenerations in primary and secondary follicles and in ovarian tissue were more common in the PRP-supplemented group. The calcium used in PRP activation was suspected to have increased the degeneration and prevented the possible positive effects of PRP. Conclusion: To the best of our knowledge, PRP-supplemented vitrification solution was used for the first time in the literature in this study in whole rat ovarian tissue vitrification. If PRP is to be used as a component in vitrification solution for rat ovarian tissue, the use of lower amounts of calcium or different methods in PRP activation, or the use of nonactivated PRP, should be considered from the beginning.

Circulating mir-200c and mir-33a may be used as biomarkers for predicting high fructose corn syrup-induced fatty liver and vitamin D supplementation-related liver changes.

BACKGROUND: Nonalcoholic fatty liver is one of the most common forms of liver disease and role of microRNAs (miRNAs) on this illness is currently unclear. It was aimed to evaluate the predictive role of circulating miR-33a and mir-200c on high fructose corn syrup (HFCS)-induced fatty liver and vitamin D3 supplementation-related hepatic changes. METHODS: Twenty-four rats were randomized into three groups: sham (n = 8), experimental fatty liver group (n = 8), and fatty liver + vitamin D3 supplementation group (n = 8). In the treatment group, 10 µg/kg/week of vitamin D3 was given by orogastric tube weekly for 4 weeks in addition to a high fructose diet. Serum AST, ALT, TNF-α, and MDA levels were tested. Liver tissue samples were examined using hematoxylin/eosin, periodic acid-Schif (PAS) and Masson's Trichrome staining. Circulating miR-33a and mir-200c were quantified by qRT-PCR method. Moreover, in silico analyses were accomplished. RESULTS: In the vitamin D3 group, results of biochemical parameters were significantly different than those of the fatty liver group (p < 0.001). Moreover, significant differences in serum levels of circulating miR-33a and mir-200c were identified among all groups (p < 0.05). Finally, more favorable histopathological changes were noticed in the vitamin D3 supplementation group. The expressions of Ki-67 were also considerably reduced in the vitamin D3 group. According to KEGG pathway analysis, mir-33a and mir-200c were found to play a common role in the Hippo signaling pathway, lysine degradation, and protein processing. DISCUSSION: Our current experimental fatty liver study showed that, in a specified dose, vitamin D3 supplementation could alleviate adverse undesirable hepatic effects of HFCS via miR-33a and mir-200c.

Evaluation of the Charlson Comorbidity Index and Laboratory Parameters as Independent Early Mortality Predictors in Covid 19 Patients.

Purpose: Various parameters have been proposed to predict the outcome of patients with coronavirus disease. The aim of this study was to evaluate the utility of the age-adjusted CCI score and biochemical parameters for predicting outcomes for COVID-19 patients on admission. Patients and methods: A total of 511 patients were included in the study. Only swab or serological tests positive patients were included. The clinical characteristics of the patients were compared between survival and non-survival COVID-19 inpatients. Hemoglobin, platelet, sedimentation, creatinine, AST, ALT, LDH, CK, albumin, ferritin, lymphocyte, neutrophil, CRP (1-5;5-10;10-20 × upper limit), procalcitonin (5-10;10-20; > 20 × upper limit), D Dimer (> 2 × upper limit), age, gender, chronic diseases and CCI scores were compared between the two groups. Results: 68 patients died and 443 patients survived. Mean age was 74.3±7.3 years in survival group and 76.7±8.0 in nonsurvival group. Age, male sex, ischemic heart disease (CHD), chronic kidney disease and active malignancy was statistically higher in non-survivor group. The biochemical parameters was compared in survival and nonsurvival group. CCI score, AST, LDH, CK, Ferritin, CRP are significantly higher and albumin, lymphocyte levels are significantly lower in nonsurvival group. D-dimer and procalcitonin levels are significantly higher in nonsurvival group. CCI score and neutrophil, creatinine, ALT, AST, d-dimer and procalcitonin elevations were correlated. Low albumin and lymphocyte levels were correlated with the CCI score. There was no significant correlation between ferritin, sedimentation, CRP levels and CCI score. A multivariate logistic regression analysis indicated that anaemia, elevated CRP (> 10-20 × upper limit), procalcitonin (> 5-10 × upper limit), ALT, AST levels and higher CCI score were independent risk factors for mortality in COVID-19 patients. Conclusion: Anaemia, elevated CRP, procalcitonin levels, ALT, AST levels and higher CCI score were found independent risk factors for mortality in COVID-19 patients.

Anticancer activity of linalool: comparative investigation of ultrastructural changes and apoptosis in breast cancer cells.

Breast cancer is the most common cancer in women in the world. Many anticancer drugs are currently used clinically have been isolated from plant species or are based on such substances. Linalool is aromatic compounds from the monoterpene group. It is the main constituents of essential oils and show antiproliferative, antioxidant, and antiseptic properties. The aim of this study was to investigate the antiproliferativeand apoptotic, effects of linalool in MCF-7 and MDA-MB-231 human breast cancer cells. MCF-7 and MDA-MB-231 human breast cancer cells were treated with different concentrations of linalool (100, 200, 400, 600, 800, 1000 µM) at 24 h and 48 h. MTT assay for cell proliferation and Annexin V assay for apoptosis was done. The morphology of breast cancer cells was investigated by light microscope and scanning electron microscope (SEM). The study show that linalool significantly induced apoptosis in all groups as dose and time-dependent (p < .05). Linalool has apoptotic and antiproliferative properties in a concentration and time-dependent manner in breast cancer cells. The cytotoxic effects of linalool on MCF-7 and MDA-MB-231 human breast cancer cells was found to be associated with apoptotic cell death. Linalool was more effective on MCF-7 human breast cancer cells in smaller amounts.

Serum albumin and FT3/FT4 ratio as additional co-morbidity parameters to predict mortality as a new approach: The Haseki Scoring Index (updated Charlson Comorbidity Index).

BACKGROUND: Charlson Comorbidity Index (CCI) is the common and valid method to predict mortality by classifying comorbidities such as cardiovascular, metabolic, renal, hepatic, pulmonary diseases, and malignancy. Novel risk factors are not included in the Charlson Comorbidity Index, such as thyroid hormone index (FT3/FT4 ratio) and serum albumin levels. In the present study, we aimed to assess whether the thyroid hormone index and albumin are useful clinical parameters in short and long-term mortality. METHODS: In the retrospective cohort study with a 5 year follow up, the data of 1292 patients who were hospitalized between January 1st-June 30th of 2014 were examined. Three months mortality as short term and 5-year mortality as long term were evaluated. RESULTS: Three months and 5 years mortality rates for 1064 patients were analyzed. We showed that hypoalbuminemia and thyroid hormone index had statistically significant effects on short and long-term mortality. According to ROC analysis it was demonstrated that the scoring system including biochemical parameters such as thyroid hormone index and serum albumin level was more significant for 3-month mortality. In addition, both scoring systems are equal in demonstrating long-term mortality. CONCLUSION: Thyroid hormone index and albumin could improve the prognostic performance of the original Charlson Comorbidity Index in short term mortality. The combined score may offer improvements in comorbidity summarization over existing scores.

Evaluation of Predisposing Metabolic Risk Factors for Portopulmonary Hypertension in Patients with NASH Cirrhosis.

PURPOSE: Metabolic parameters are important for the development of portopulmonary hypertension (PoPH) during nonalcoholic steatohepatitis (NASH)-associated cirrhosis. This study evaluated patients with NASH-associated cirrhosis to determine metabolic risk factors for portopulmonary hypertension. PATIENTS AND METHODS: Data on 171 patients (120 men and 51 women) with NASH-associated cirrhosis who were seen in Florence Nightingale Hospital's gastroenterology Clinic from 2009 to 2018 was obtained from the Hospital database. A pulmonary artery systolic pressure >35 mmHg was defined as PH (pulmonary hypertension) according to standard transthoracic echocardiography. Portal hypertension was diagnosed from clinical symptoms and dilated portal veins shown by abdominal ultrasound or computed tomography (CT). Pulmonary patients with portal hypertension were diagnosed with portopulmonary hypertension (PoPH). RESULTS: A total of 171 patients with NASH-associated cirrhosis were included in this study. Of these, 43 patients had PoPH. These patients had increased TSH (p=0.004), bilirubin (p=0.023) and triglyceride (p=0.048) levels, higher MELD scores (p=0.018) and decreased hemoglobin (p=0.05). MELD score and hemoglobin, total bilirubin, TSH, and triglyceride levels were all included in a multivariate logistic regression model and TSH levels were independently associated with increased risk of PoPH. CONCLUSION: Increased TSH is an independent risk factor for PoPH.

Multi-parametric evaluation of autologous cultivated Limbal epithelial cell transplantation outcomes of Limbal stem cell deficiency due to chemical burn.

BACKGROUND: The sparsity of established tools for the grading of limbal stem cell deficiency hinder objective assessments of the clinical outcome of cultivated limbal epithelial cell transplantation. To advance towards the development of standards for the comparison of the outcomes of these bio-surgical protocols we have now applied a battery of recognized objective and patient-declared subjective outcome criteria to the autologous modality of cultivated limbal epithelial cell transplantation. METHODS: The prospective study involved ten patients (M/F = 9/1; mean age = 42.1 years) displaying overt unilateral limbal stem cell deficiency complying with the inclusion criteria described in Methods. Limbal biopsies were obtained from the contralateral eye and their outgrowths after 2-week cultures were transplanted on the affected eye after pannus resection. Outcomes were followed up for 12 months. The objective tests were scores for best-corrected visual acuity (BCVA); using the LogMAR scale, a multiparametric ocular surface score (OSS), and the Schirmer's test. Subjective scores were based on patient answers to a) perception of visual improvement/pain; b) the 25-item National Eye Institute Visual Function Questionnaire (NEI-VFQ 25); and c) the 12-item Ocular Surface Disease Index Questionnaire (OSDI). All procedures were performed under good manufacture practices using solely xeno-free reagents. In all cases, a single biopsy was divided into two pieces and they were expanded in order to prevent outgrowth failure. In 5 patients, both biopsies generated healthy culture sheet. In those cases the lesser outgrowth were used for immune-histological characterization. RESULTS: The experimental parallel outgrowth samples showed a similar percent of p63α+ cells. PreOp and 12-month PostOp BCVAs and OSSs were, respectively, 1.15 ± 0.70; 0.21 ± 0.13 and 7.40 ± 2.01; 2,30 ± 1.30, (p < 0.05). Patient's responses to all three question sets except ocular pain were consistent with significant improvement (p < 0.05). CONCLUSION: Objective clinical metrics demonstrate that in patients with limbal stem cell deficiency, cultivated limbal epithelial cell transplantation improves vision and ocular surface health and subjective visual perceptions.

The healing effects of Hyperium perforatum (St. John's Wort) on experimental alkaline corrosive eosephageal and stomach burns.

BACKGROUND: The most frequent etiologic cause is alkaline substances. We investigated the protective effects of the plant St. John 's Wort (Hypericum perforatum). METHODS: We included 42 Wistar albino rats weighing between 200-300 grams and divided into six groups as Group 1: Control, Group 2: Burn+Saline (BS), Group 3: Burn+St. John's Wort (BSJW), Group 4: Burn+Plasebo (BP), Group 5: St. John's Wort (SJW), Group 6: Placebo (P). After 15 days of treatment, esophagus, stomach and liver tissue samples were derived by dissection for histopathologic and biochemical markers. The cytotoxic effects of formulation on fibroblasts is evaluated in vitro on human dermoblast fibroblast line (HDFa, Gibco Invitrogen cell culture, C-013-5C). RESULTS: The weight of the rats increased in Group 1, 3, 4, 6, decreased in Group 2 and did not change in Group 5. In the BSJW group, submucosal collagen accumulation, muscularis mucosa damage, tunica muscularis damage and collagen accumulation in esophagus were similar to the control group but lesser than BS and placebo group. In the stomach, mucosal damage, gastric gland dilatation, submucosal polymorphonuclear infiltration were similar to the control group and lesser than the BS group. The lethal concentration of SJW was 2.58 gr/mL. CONCLUSION: SJW substrate is effective in protecting the esophagus and stomach in mild to moderate alcali corrosive burns in the subacute period. We should keep in mind the protective effects of STW substrate in alkaline corrosive burns of the gastrointestinal system.

Comparison of ultrastructural changes and the anticarcinogenic effects of thymol and carvacrol on ovarian cancer cells: which is more effective?

Ovarian cancer is the seventh most common cancer worldwide in women. Many anticancer drugs are currently used clinically have been isolated from plant species or are based on such substances. Thymol (5-methyl-2-isopropylphenol) and carvacrol are oxygenated aromatic compounds from the monoterpene group. They are the main constituents of thyme essential oil and show antiproliferative, antioxidant, and antiseptic properties. The aim of this study is to compare the antiproliferative and apoptotic effects of thymol and carvacrol on SKOV-3 ovarian cancer cell line. The cancer cells were treated with different concentrations of thymol and carvacrol (100, 200, 400, 600 µM) at 24 h and 48 h durations. The cell viability was investigated by MTT assay and analysis of apoptosis with annexin V assay was determined. The study show that thymol and carvacrol significantly induced apoptosis in all groups as dose and time-dependent (p < .05). The data in the present study demonstrated that thymol and carvacrol have apoptotic and antiproliferative properties in a concentration-dependent manner toward ovarian cancer cells. SKOV-3 cancer cell line was much more sensitive to the toxic effect of thymol than carvacrol.

Urotensin receptor antagonist palosuran attenuates cyclosporine-a-induced nephrotoxicity in rats.

BACKGROUND: Cyclosporine-A (CsA) is widely used for immunosuppressive therapy in renal transplantation. Nephrotoxicity is the main dose-limiting undesirable consequence of CsA. Urotensin II (U-II), a novel peptide with a powerful influence on vascular biology, has been added to the list of potential renal vascular regulators. Upregulation of the urotensin receptors and elevation of plasma U-II levels are thought to possibly play a role in the etiology of renal failure. OBJECTIVES: The present study examines this hypothesis by evaluating renal function and histology with regard to the potential role of U-II and its antagonist, palosuran, in the pathogenesis of CsA-induced nephrotoxicity in rats. MATERIAL AND METHODS: Male Sprague-Dawley rats were treated with CsA (15 mg/kg, for 21 days, intraperitoneally) or CsA + palosuran (300 mg/kg, for 21 days). Renal function was measured and histopathology, U-II immunostaining and protein detection with western blotting of the kidneys were performed. RESULTS: Cyclosporine-A administration caused a marked decline in creatinine clearance (Ccr). Fractional sodium excretion (FENa) tended to increase in the CsA-treated rats. Plasma U-II levels decreased in the CsA-treated rats. Cyclosporine-A treatment resulted in a marked deterioration in renal histology and an increase in the expression of U-II protein in the kidneys. Palosuran's improvement of renal function manifested as a significant decrease in serum creatinine levels and a significant increase in urine creatinine levels, resulting in a marked increase in Ccr. Palosuran produced a significant normalization of kidney histology and prevented an increase in U-II expression. CONCLUSIONS: Cyclosporine-A-induced renal impairment was accompanied by an increase in U-II expression in kidneys and a contrary decrease in systemic U-II levels. Palosuran improved the condition of rats suffering from renal dysfunction by preventing the decrease in renal U-II expression without affecting the systemic levels of U-II. The protective effect of palosuran in CsA nephrotoxicity is possibly independent of its U-II receptor antagonism.

Who needs extra time? Amniotic membrane wrapped pauci-suture model for rapid anastomoses.

An ideal anastomosis method will obtain the highest post-anastomotic vessel patency and will repair the vessel anatomically with minimal thrombosis in an easier, faster and cheaper fashion. To achieve these goals an anastomosis model using an amniotic membrane is introduced. The study was performed on the femoral arteries of 22 Wistar Albino rats (11 control group, 11 experimental group). In the experiment group, the microvascular anastomosis was completed with three sutures and a patch of amniotic membrane which was wrapped around the anastomotic site. The conventional anastomosis technique with eight sutures was performed in the control group. The effects of the model on the patency and histological structure of the vessels were evaluated. As a result, normal patency was determined radiologically and macroscopically in all of the anastomoses. No thrombosis or aneurysm was detected in any of the anastomoses. In the angiographic study, vessel patency was detected in both the control and experimental groups. The average time to complete the arterial anastomosis was 18.14 (±2.84) and 10.39 (±2.45) minutes in the control and the experiment groups respectively. In the histological studies, anti-eNOS staining revealed that endothelin levels were significantly higher in the experimental group. This method describes a new anastomosis model in microvascular surgery with promising results that call for additional experimental studies and further clinical implementations. We believe that this experimental technique can be put into clinical practice as an alternative to the conventional microvascular anastomosis technique.

Treatment with NADPH oxidase inhibitor apocynin alleviates diabetic neuropathic pain in rats.

Increased reactive oxygen species by the activation of NADPH oxidase (NOX) contributes to the development of diabetic complications. Apocynin, a NOX inhibitor, increases sciatic nerve conductance and blood flow in diabetic rats. We investigated potential protective effect of apocynin in rat diabetic neuropathy and its precise mechanism of action at molecular level. Rat models of streptozotocin-induced diabetes were treated with apocynin (30 and 100 mg/kg per day, intragastrically) for 4 weeks. Mechanical hyperalgesia and allodynia were determined weekly using analgesimeter and dynamic plantar aesthesiometer. Western blot analysis and histochemistry/immunohistochemistry were performed in the lumbar spinal cord and sciatic nerve respectively. Streptozotocin injection reduced pain threshold in analgesimeter, but not in aesthesiometer. Apocynin treatment increased pain threshold dose-dependently. Western blot analysis showed an increase in catalase and NOX-p47phox protein expression in the spinal cord. However, protein expressions of neuronal and inducible nitric oxide synthase (nNOS, iNOS), superoxide dismutase, glutathion peroxidase, nitrotyrosine, tumor necrosis factor-α, interleukin-6, interleukin-1ß, aldose reductase, cyclooxygenase-2 or MAC-1 (marker for increased microgliosis) in the spinal cord remained unchanged. Western blot analysis results also demonstrated that apocynin decreased NOX-p47phox expression at both doses and catalase expression at 100 mg/kg per day. Histochemistry of diabetic sciatic nerve revealed marked degeneration. nNOS and iNOS immunoreactivities were increased, while S-100 immunoreactivity (Schwann cell marker) was decreased in sciatic nerve. Apocynin treatment reversed these changes dose-dependently. In conclusion, decreased pain threshold of diabetic rats was accompanied by increased NOX and catalase expression in the spinal cord and increased degeneration in the sciatic nerve characterized by increased NOS expression and Schwann cell loss. Apocynin treatment attenuates neuropathic pain by decelerating the increased oxidative stress-mediated pathogenesis in diabetic rats.

Antagonistic Effect of Oxytocin and Tacrolimus Combination on Adipose Tissue - Derived Mesenchymal Stem Cells: Antagonistic effect of oxytocin and tacrolimus.

Tacrolimus (FK506) is a chemotherapeutic agent, which uses calcineurin pathway via inhibiting the stimulation of T cells to prevent the formation of immune response in the recipient individual in organ transplants. FK506 is mainly metabolized in the liver by cytochrome P450 enzyme system and is known that it has high toxic effects on different cells. Mesenchymal stem cells (MSCs) have recently gained importance since their potential to be used in cellular therapy and tissue regeneration. In some clinical cases, MSCs are transferred into the patient after the organ transplantations in order to support the treatment. Because of their immunomodulatory actions and assistance to the regeneration, popularity of MSCs have been increasing recently. However, since immunosuppressive agents have a potential cytotoxic and apoptotic effect on MSCs, researches have attempted to use it as a combination with an agent that alleviates these effects. Oxytocin (OT) is primarily acting as a neuromodulator in humans and is a peptide hormone secreted by the pituitary gland of the neurohypophysis. OT has such effects on cells as to confer resistance against oxidative stress on cells and to increase the proliferation and help regeneration. Studies on the active substance of FK506 were aimed to investigate the cytotoxic effects on human adipose tissue derived MSCs. The purpose of this study was to determine the cytotoxic, apoptotic and morphological effects of FK506, an immunosuppressive agent, on adipose tissue - derived MSC (ADMSC) which has the potential to be used for immune suppression. In addition, it was aimed to determine whether the agent could reduce the cytotoxic, apoptotic, and morphological effects on ADMSCs when used in combination with OT. For this purpose, the cytotoxic effects of the FK506 and OT on ADMSCs were determined by time and dose dependent manner by the WST-1 test. Isobologram analysis was evaluated using the WST-1 test according to IC50 values of FK506 and OT. The apoptotic effects of the agents on the ADMSCs were determined by the Annexin V method. Immunofluorescence staining was performed to determine morphological changes. Changes in the levels of oxidative stress markers were measured by colorimetric and flourometric methods using lipid peroxidation, superoxide dismutase activity, catalase activity and glutathione peroxidase assays. The IC50 values of FK506 and OT on ADMSCs were calculated as 17.44µM and 13.43µM, respectively.FK506 and OT were found to have antagonistic activity on ADMSCs (CI value of the combination was 1.24). The effects of the agents individually and in combination on the levels of apoptosis and oxidative stress markers have been evaluated. When the results obtained from the study are evaluated, the adipose- tissue derived mesenchymal stem cells used with takrolimus and oxytocin combination have a potential for novel treatment approaches.

Limbal Stem Cell Deficiency and Treatment with Stem Cell Transplantation.

The cornea is the outermost tissue of the eye and it must be transparent for the maintenance of good visual function. The superficial epithelium of the cornea, which is renewed continuously by corneal stem cells, plays a critical role in the permanence of this transparency. These stem cells are localized at the cornea-conjunctival transition zone, referred to as the limbus. When this zone is affected/destroyed, limbal stem cell deficiency ensues. Loss of limbal stem cell function allows colonization of the corneal surface by conjunctival epithelium. Over 6 million people worldwide are affected by corneal blindness, and limbal stem cell deficiency is one of the main causes. Fortunately, it is becoming possible to recover vision by autologous transplantation of limbal cells obtained from the contralateral eye in unilateral cases. Due to the potential risks to the donor eye, only a small amount of tissue can be obtained, in which only 1-2% of the limbal epithelial cells are actually limbal stem cells. Vigorous attempts are being made to expand limbal stem cells in culture to preserve or even enrich the stem cell population. Ex vivo expanded limbal stem cell treatment in limbal stem cell deficiency was first reported in 1997. In the 20 years since, various protocols have been developed for the cultivation of limbal epithelial cells. It is still not clear which method promotes effective stem cell viability and this remains a subject of ongoing research. The most preferred technique for limbal cell culture is the explant culture model. In this approach, a small donor eye limbal biopsy is placed as an explant onto a biocompatible substrate (preferably human amniotic membrane) for expansion. The outgrowth (cultivated limbal epithelial cells) is then surgically transferred to the recipient eye. Due to changing regulations concerning cell-based therapy, the implementation of cultivated limbal epithelial transplantation in accordance with Good Laboratory Practice using xenobiotic-free systems is becoming widely accepted both in Turkey and worldwide.

Neuroprotective Effects of Eexenatide in a Rotenone-Induced Rat Model of Parkinson's Disease.

BACKROUND: Several studies suggest an association between Parkinson's disease (PD) and type 2 diabetes mellitus; these 2 diseases are both known to affect the common molecular pathways. As a synthetic agonist for the glucagon-like peptide 1 receptor, exenatide has been evaluated as a neuroprotective agent in multiple animal models. Rotenone models of PD have great potential for the investigation of PD pathology and motor and nonmotor symptoms, as well as the role of gene-environment interactions in PD causation and pathogenesis. Therefore, in this study, the neurochemical, behavioral and histologic effects of exenatide on a rotenone-induced rat model of PD were examined. MATERIALS AND METHODS: Eighteen adult male rats were randomly divided into the following 3 groups (n = 6): 1 group received stereotaxical infusion of dimethyl sulfoxide (vehicle, group 1) and the others received stereotaxical infusion of rotenone (groups 2 and 3). Apomorphine-induced rotation test was applied to the rats after 10 days. Thereafter, group 2 was administered isotonic saline, whereas group 3 was administered exenatide for 28 days. RESULTS: Malondialdehyde and tumor necrosis factor alpha levels increased in the rats with PD induced by rotenone, whereas malondialdehyde and tumor necrosis factor alpha levels markedly decreased in the rats treated with exenatide. The apomorphine-induced rotation test scores of exenatide-treated rats were determined to be lower compared with the untreated group. Additionally, treatment with exenatide significantly reduced the loss of dopaminergic neurons in striatum. CONCLUSIONS: These results have shown that exenatide has neuroprotective, anti-inflammatory and antioxidant effects in a rotenone-induced rat model of PD.

Useful Effects of Melatonin in Peripheral Nerve Injury and Development of the Nervous System.

This review summarizes the role of melatonin (MLT) in defense against toxic-free radicals and its novel effects in the development of the nervous system, and the effect of endogenously produced and exogenously administered MLT in reducing the degree of tissue and nerve injuries. MLT was recently reported to be an effective free radical scavenger and antioxidant. Since endogenous MLT levels fall significantly in senility, these findings imply that the loss of this antioxidant could contribute to the incidence or severity of some age-related neurodegenerative diseases. Considering the high efficacy of MLT in overcoming much of the injury not only to the peripheral nerve but also to other organs, clinical trials for this purpose should be seriously considered.

Neuroprotective effects of erythropoietin on Alzheimer's dementia model in rats.

BACKGROUND: Although Alzheimer's disease (AD) is the most common age-related neurodegenerative disease and characterized by memory impairment, only symptomatic treatments are available. OBJECTIVES: Because recombinant human erythropoietin (rhEPO) has various neuroprotective effects and improves cognitive function in animal models of neurodegenerative disorders, we investigated the therapeutic effects of rhEPO in an intracerebroventricular (ICV)-streptozotocin (STZ) animal model of sporadic-AD. MATERIAL AND METHODS: A total of 24 Sprague-Dawley adult rats were divided into 4 groups of naive control (n = 6), sham-operated (n = 6), ICV-STZ + saline (n = 6) and ICV-STZ + rhEPO (n = 6). Twelve rats with Alzheimer's disease, induced by STZ injection (3 mg/kg) into both lateral ventricles using a stereotaxic frame (bilaterally ICV-STZ), were divided into 2 groups 5 days after the STZ injection: one treated with rhEPO 5000 (IU/kg/day, i.p.) and the other with 0.9% NaCl (1 mL/kg/day, i.p.) for 2 weeks. The sham-operated rats received bilaterally ICV-0.9% NaCl. No surgical operation or treatment was given to the naive-control animals. On day 20, a passive avoidance learning (PAL) test was used followed by sacrification and removal of the brain tissue in all animals. Brain TNF-α and ChAT levels were determined, and neurons in the hippocampal CA1 and CA3 regions were counted by Cresyl violet staining. RESULTS: ICV-STZ was found to significantly shorten the latency time on the PAL, increase brain TNF-α level, and decrease brain ChAT activity and the number of neurons in the hippocampal CA1 and CA3 regions. On the other hand, rhEPO significantly attenuated all these detrimental effects induced by STZ. CONCLUSIONS: RhEPO treatment significantly prevented the ICV-STZ-induced memory deficit by attenuating the hippocampal neuronal loss, neuroinflammation and cholinergic deficit in rats. This result suggests that rhEPO may be beneficial for treating AD.

Effect of platelet-rich plasma on reconstruction with nerve autografts.

Despite advances in understanding of peripheral nerve injuries and regeneration and advances in surgical techniques, successful outcomes cannot be guaranteed after reconstructive surgery. Platelet-rich plasma (PRP) has been reported to have positive effects on nerve regeneration, as well as on tissue healing. The present study was designed to evaluate the effect of PRP on nerve-grafted defects. Sprague-Dawley rats were divided into four surgery groups (n=7 in each). A 1-cm long nerve defect was created in the upper thigh and then reconstructed using a nerve autograft in all groups. The wet muscle weights, electromyographic findings, and histomorphologic changes were evaluated 10 weeks later. As shown by both the electromyographic (p<0.001) and histomorphologic findings (p<0.001), PRP had more positive effects on nerve gap reconstruction in Group 3 then Group 4 as compared to the control groups. The present study is novel in that it evaluated the regeneration effect of PRP on a large nerve defect reconstructed with a nerve graft rather than primary repair. The results are encouraging for further experimental studies on the role of PRP in nerve healing.

Clinical Use and Patentability of Cord Blood.

BACKGROUND: The blood in the umbilical cord that provides the connection between mother and fetus during pregnancy is called cord blood. The blood of umbilical cord which is usually got rid of following birth, is a very rich stem cell source. OBJECTIVE: Cord blood collection gives no harm to the mother and baby. Besides, its allogeneic and autologous usage, the most important disadvantage is that the number of cells is insufficient in adults. CONCLUSION: Today, it is predominantly used for therapeutic purposes for many diseases. The aim of this review is giving a detailed information about groups of stem cells in cord blood and determining the point of clinical use.