The Effects of Apocynin on Monosodium Glutamate Induced Liver Damage of Rats.

Monosodium glutamate (MSG) is found in refined foods. Apocynin (APO) is a selective NADPH oxidase (NOX) inhibitor. The aim of this experimental study was to investigate possible effects of MSG and the curative effects of APO in rats. Twenty-eight male Sprague-Dawley rats were randomly divided into four groups (Normal control, APO, MSG and MSG + APO, n:7 for each group). The MSG and MSG + APO groups received 120 mg/kg MSG solution orally for 28 consecutive days. The APO and MSG + APO groups received 25 mg/kg APO solution orally for 5 days until the end of the experiment. At the end of the experiment, all rats were sacrificed and liver tissue and blood samples were taken for histological, ultrastructural, and biochemical analyses. In the MSG group, vacuolization and loss in glycogen content in the hepatocytes, leukocyte infiltration and fibrosis in the liver parenchyme and portal triads, were observed. Terminal deoxynucleotidyl transferase dUTP (TUNEL)-positivity and NADPH oxidase (NOX)-2-positivity were higher in the MSG group compared with the other experimental groups. The concentrations of alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), total bilirubin, malondialdehyde (MDA), and myeloperoxidase (MPO) were higher, whereas albumin, glutathione (GSH), and superoxide (SOD) levels were lower in the MSG group. All these data has been reversed in MSG + APO group. The histological and biochemical criteria indicated the prominent ameliorating effect of APO on MSG -induced liver injury.

Apocynin Ameliorates Monosodium Glutamate Induced Testis Damage by Impaired Blood-Testis Barrier and Oxidative Stress Parameters.

BACKGROUND: the aim of this study was to investigate the effects of apocynin (APO) on hormone levels, the blood-testis barrier, and oxidative biomarkers in monosodium glutamate (MSG) induced testicular degeneration. METHODS: Sprague Dawley male rats (150-200 g; n = 32) were randomly distributed into four groups: control, APO, MSG, and MSG + APO. MSG and MSG + APO groups were administered MSG (120 mg/kg) for 28 days. Moreover, the APO and MSG + APO groups received APO (25 mg/kg) during the last five days of the experiment. All administrations were via oral gavage. Finally, biochemical analyses were performed based on the determination of testosterone, follicle-stimulating hormone (FSH), luteinizing hormone (LH), malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD), as well as light and transmission electron microscopic examinations, assessment of sperm parameters, ZO-1, occludin, NOX-2, and TUNEL immunohistochemistry were evaluated. RESULTS: MSG increased both the oxidative stress level and apoptosis, decreased cell proliferation, and caused degeneration in testis morphology including in the blood-testis barrier. Administration of apocynin reversed all the deteriorated morphological and biochemical parameters in the MSG + APO group. CONCLUSIONS: apocynin is considered to prevent testicular degeneration by maintaining the integrity of the blood-testis barrier with balanced hormone and oxidant/antioxidant levels.

Ferulic Acid Treats Gastric Ulcer via Suppressing Oxidative Stress and Inflammation.

(1) Background: The aim of the present study was to evaluate the gastroprotective potential of ferulic acid (FA) on indomethacin-induced gastric ulcers in rats with macroscopic and microscopic examinations along with biochemical assays. (2) Methods: After 24 h starvation, the ulcer was induced in male Sprague-Dawley rats by subcutaneous indomethacin (25 mg/kg) injection. Fifteen minutes after ulcer induction, rats were treated with either tween 80 or FA. FA was given by oral gavage at 100 mg/kg, 250 mg/kg, and 500 mg/kg. In the fourth hour, rats were euthanized and collected gastric samples were evaluated macroscopically and microscopically. Antioxidant parameters including malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), and inflammatory parameters comprising of myeloperoxidase (MPO), Tumor Necrosis Factor (TNF)-α, Interleukin (IL)-1ß, IL-6 and Nuclear Factor Kappa-B (NF-κB) p65 levels were also determined. (3) Results: Indomethacin injection significantly increased the macroscopic and microscopic scores. In addition, it increased the gastric MDA, MPO, TNF-α, IL-1ß, IL-6, and NF-κB p65 levels but reduced SOD and GSH content. Treatment with FA significantly improved the gastric injury macroscopically and microscopically. Moreover, FA displayed a marked decrease in the gastric levels of MDA, MPO, TNF-α, IL-1ß, IL-6, and NF-κB p65 and a significant increase in SOD and GSH compared to the INDO group. Ultimately, 250 mg/kg FA was determined as the most effective dose. (4) Conclusion: Our results revealed that FA has a gastroprotective effect against indomethacin-induced gastric ulcers in rats due to its antioxidant and anti-inflammatory properties. As a result, FA may be a potential treatment choice for gastric ulcers.

Ferulic acid attenuates pancreaticobiliary duct occlusion-induced inflammation in both pancreas and liver.

INTRODUCTION: Acute pancreatitis is a systemic inflammatory disorder characterized by the hyperactivation of digestion enzymes and the release of proinflammatory cytokines. Ferulic acid (FA) is a hydroxycinnamic acid derivative that has recently been shown to have antioxidant and anti-inflammatory properties. AIM: The anti-inflammatory effects of FA were investigated in the pancreaticobiliary duct ligation (PBDL)-induced pancreatitis model. METHODS: Wistar albino rats (250-300 g; female = male) were divided into sham operation and PBDL groups. Some PBDL-performed animals were given intragastric saline or 250 mg/kg FA or 500 mg/kg FA 30 min before the PBDL and for 3 consecutive days. Moreover, the control group received saline. Blood samples are collected at the 24th, 48th, and 72nd hours to measure serum tumor necrosis factor (TNF)-α, liver, and pancreatic enzymes. At the 72nd hour, rats were euthanized; pancreas, lung, and liver samples were collected, scored microscopically, and analyzed for myeloperoxidase activity, malondialdehyde, and glutathione levels. One-way ANOVA with Tukey-Kramer tests were used for statistical analysis. RESULTS: FA treatment reduced myeloperoxidase activity and prevented the depletion of glutathione in all three tissues. With FA treatments, high malondialdehyde levels in the pancreas and liver were reduced, as were serum TNF- α, amylase, lipase, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels. Additionally, FA ameliorated microscopic damage in the pancreas and liver significantly. CONCLUSION: According to the findings, FA protects endogenous antioxidant content, prevents neutrophil infiltration, and decreases lipid peroxidation in PBDL-induced pancreatitis. Furthermore, FA improves tissue damage induced by pancreatitis with its anti-inflammatory effects.

Loop nerve graft prefabrication for peripheral nerve defect reconstruction.

BACKGROUND: Delayed autologous nerve graft reconstruction is inevitable in devastating injuries. Delayed or prolonged repair time has deleterious effects on nerve grafts. We aimed improving and accelerating nerve graft reconstruction process in a rat long nerve defect model with loop nerve graft prefabrication particularly to utilize for injuries with tissue loss. METHODS: Twenty-four Sprague-Dawley rats were allocated into three groups. 1.5 cm long peroneal nerve segment was excised, reversed in orientation, and used as autologous nerve graft. In conventional interpositional nerve graft group (Group 1), nerve defects were repaired in single-stage. In loop nerve graft prefabrication group (Group 2), grafts were sutured end-to-end (ETE) to the proximal peroneal nerve stumps. Distal ends of the grafts were sutured end-to-side to the peroneal nerve stumps 5 mm proximal to the ETE repair sites in first stage. In second stage, distal ends of the prefabricated grafts were transposed and sutured to distal nerve stumps. In staged conventional interpositional nerve graft group (Group 3), grafts were sutured ETE to proximal peroneal nerve stumps in first stage. Distal ends of the grafts and nerve stumps were tacked to the surrounding muscles until the final repair in second stage. Follow-up period was 4 weeks for each stage in Groups 2 and 3, and 8 weeks for Group 1. Peroneal function index (PFI), electrophysiology, and histological assessments were conducted after 8 weeks. P<0.05 was considered significant for statistical analysis. RESULTS: PFI results of Group 1 (-22.75±5.76) and 2 (-22.08±6) did not show statistical difference (p>0.05). Group 3 (-33.64±6.4) had a statistical difference compared to other groups (p<0.05). Electrophysiology results of Group 1 (16.19±2.15 mV/1.16±0.21 ms) and 2 (15.95±2.82 mV/1.17±0.16 ms) did not present statistical difference (p>0.05), whereas both groups had a statistical difference compared to Group 3 (10.44±1.96 mV/1.51±0.15 ms) (p<0.05). Axon counts of Group 1 (2227±260.4) and 3 (2194±201.1) did not have statistical difference (p>0.05), whereas both groups had significantly poor axon counts compared to Group 2 (2531±91.18) (p<0.05). CONCLUSION: Loop nerve graft prefabrication improved axonal regeneration without delay. Loop prefabrication can accelerate prolonged regeneration time for the injuries indicating a delayed nerve reconstruction. Higher axon counts derived with loop nerve prefabrication may even foster its investigation in immediate long nerve defect reconstructions in further studies.

Apocynin reduces cytotoxic effects of monosodium glutamate in the brain: A spectroscopic, oxidative load, and machine learning study.

Herein, we examined the modulatory effects ofApocynum (APO) on Monosodium Glutamate (MSG)-induced oxidative damage on the brain tissue of rats after long-term consumption of blood serum components by biochemical assays, Fourier transform infrared spectroscopy(FTIR), and machine learning methods. Sprague-Dawley male rats were randomly divided into the Control, Control + APO, MSG, and MSG + APO groups (n = 8 per group). All administrations were made by oral gavage saline, MSG, or APO and they were repeated for 28 days of the experiments. Brain tissue and blood serum samples were collected and analyzed for measurement levels ofmalondialdehyde (MDA),glutathione (GSH),myeloperoxidase (MPO), superoxide dismutase (SOD) activity, and Spectroscopic analysis. After 29 days, the results were evaluated using machine learning (ML). The levels of MDA and MPO showed changes in the MSG and MSG + APO groups, respectively. Changes in the proteins and lipids were observed in the FTIR spectra of the MSG groups. Additionally, APO in these animals improved the FTIR spectra to be similar to those in the Control group. The accuracy of the FTIR results calculated by ML was 100%. The findings of this study demonstrate that Apocynin treatment protectsagainst MSG-induced oxidative damage by inhibitingreactive oxygen speciesand upregulatingantioxidant capacity, indicating its potential in alleviatingthe toxic effects of MSG.

Hair diameter measurements for planning follicular unit extraction surgery (FUE): Is there a correlation between the micrometer caliper and scanning electron microscopy (SEM) findings?

BACKGROUND: Hair shaft diameter is one of the most important factors for the outcome of follicular unit extraction (FUE) surgery. In fact, the hair shaft is elliptical. Therefore, it has a long and short axis. Many hair transplantation surgeons use manual micrometer caliper for gauging hair diameter and use the results in the management of recipient and donor area. AIM: In this study, we aimed to identify the dependability of micrometer caliper and also the hair diameter diversity pattern in the donor area. PATIENTS/METHODS: Two hundred and seventy hairs were collected from three males with androgenetic alopecia. Hair samples were obtained from the 1 cm2 boxes from superior to inferior at the mid-point of temporal, parietal, and occipital donor areas. The diameter of each hair was measured both with a micrometer caliper and scanning electron microscopy (SEM). RESULTS: Average diameter measured by scanning electron microscopy was 83.01 µm for the long axis and 51.51 µm for the short axis. The average value for the micrometer caliper measurement was 53.32 µm. Comparison of micrometer caliper results with the short-axis measurements of SEM revealed a strong significant correlation. The hair diameters from superior, middle, and inferior boxes revealed a tendency to decrease toward the inferior regions. CONCLUSIONS: A manual micrometer caliper is a dependable tool for planning FUE surgery, and it measures the short axis of the elliptical hair shaft. Hair diameter tends to decrease toward the inferior regions of the donor area.

Gamma-irradiated SARS-CoV-2 vaccine candidate, OZG-38.61.3, confers protection from SARS-CoV-2 challenge in human ACEII-transgenic mice.

The SARS-CoV-2 virus caused the most severe pandemic around the world, and vaccine development for urgent use became a crucial issue. Inactivated virus formulated vaccines such as Hepatitis A and smallpox proved to be reliable approaches for immunization for prolonged periods. In this study, a gamma-irradiated inactivated virus vaccine does not require an extra purification process, unlike the chemically inactivated vaccines. Hence, the novelty of our vaccine candidate (OZG-38.61.3) is that it is a non-adjuvant added, gamma-irradiated, and intradermally applied inactive viral vaccine. Efficiency and safety dose (either 1013 or 1014 viral RNA copy per dose) of OZG-38.61.3 was initially determined in BALB/c mice. This was followed by testing the immunogenicity and protective efficacy of the vaccine. Human ACE2-encoding transgenic mice were immunized and then infected with the SARS-CoV-2 virus for the challenge test. This study shows that vaccinated mice have lowered SARS-CoV-2 viral RNA copy numbers both in oropharyngeal specimens and in the histological analysis of the lung tissues along with humoral and cellular immune responses, including the neutralizing antibodies similar to those shown in BALB/c mice without substantial toxicity. Subsequently, plans are being made for the commencement of Phase 1 clinical trial of the OZG-38.61.3 vaccine for the COVID-19 pandemic.

Preclinical efficacy and safety analysis of gamma-irradiated inactivated SARS-CoV-2 vaccine candidates.

COVID-19 outbreak caused by SARS-CoV-2 created an unprecedented health crisis since there is no vaccine for this novel virus. Therefore, SARS-CoV-2 vaccines have become crucial for reducing morbidity and mortality. In this study, in vitro and in vivo safety and efficacy analyzes of lyophilized vaccine candidates inactivated by gamma-irradiation were performed. The candidate vaccines in this study were OZG-3861 version 1 (V1), an inactivated SARS-CoV-2 virus vaccine, and SK-01 version 1 (V1), a GM-CSF adjuvant added vaccine. The candidate vaccines were applied intradermally to BALB/c mice to assess toxicity and immunogenicity. Preliminary results in vaccinated mice are reported in this study. Especially, the vaccine models containing GM-CSF caused significant antibody production with neutralization capacity in absence of the antibody-dependent enhancement feature, when considered in terms of T and B cell responses. Another important finding was that the presence of adjuvant was more important in T cell in comparison with B cell response. Vaccinated mice showed T cell response upon restimulation with whole inactivated SARS-CoV-2 or peptide pool. This study shows that the vaccines are effective and leads us to start the challenge test to investigate the gamma-irradiated inactivated vaccine candidates for infective SARS-CoV-2 virus in humanized ACE2 + mice.

Caffeic acid attenuates gastric mucosal damage induced by ethanol in rats via nitric oxide modulation.

Anti-oxidant and anti-inflammatory properties of caffeic acid (CA) have been reported recently. In this study, the therapeutic effects of CA on ethanol-induced ulcer and the roles of nitric oxide and cholinergic pathways in these effects were investigated. Ulcer was induced by ethanol via oral gavage. Ulcer induced rats were treated with either vehicle (ulcer group) or CA (100, 250 or 500 mg/kg, per oral gavage). Macroscopic evaluation showed that 250 mg/kg CA was the effective dose. To elucidate the action mechanism of CA, 10 mg/kg l-NAME or 1 mg/kg atropine sulfate was administered to 250 mg/kg CA treated groups. All rats were decapitated 1 h after ulcer induction and gastric samples were scored macroscopically and microscopically, and analyzed for myeloperoxidase (MPO), malondialdehyde (MDA), and glutathione (GSH) levels. ANOVA test was used for statistical analyses. Macroscopic and microscopic damage scores, MDA levels and MPO activity were increased while GSH levels were decreased in ulcer group. Treatment with 250 mg/kg and 500 mg/kg CA reduced macroscopic and microscopic damage scores, decreased MPO activity and MDA levels, and preserved the depleted glutathione significantly. l-NAME administration before CA treatment elevated MDA levels, MPO activity and depleted glutathione. However, atropine sulfate had no effect on biochemical parameters. We conclude that CA ameliorates ethanol-induced gastric mucosal damage, and NO pathway contributes to this effect. On the other hand, there is a lack of evidence for the contribution of the muscarinic cholinergic system.

Morphological evaluation of the effects of exercise on high-fat-diet-induced liver damage in rats.

BACKGROUND/AIMS: This study was aimed to investigate the protective effects of swimming exercise on nonalcoholic fatty liver disease (NAFLD) associated with high fat diet-induced obesity, using microscopical and biochemical parameters. MATERIALS AND METHODS: Sprague Dawley male rats were fed either standard chow (STD group; 6% fat) or high-fat diet (HFD group; 45% fat) for 18 weeks. Animals were divided into four groups, STD, STD + EXC, HFD, HFD + EXC. Exercise groups were submitted to swimming training 5 days of week and 1h of per day, during the last 6 weeks of the experiment. At the end of the experiment, liver samples were evaluated for morphologically and ultrastructurally. Moreover, malondialdehyde (MDA) and glutathione (GSH) levels were evaluated in liver samples. RESULTS: Normal morphology of liver parancyma with hepatocytes and sinusoids was observed in the STD and STD+EXC groups. Steatosis, lipid accumulation, ballooned hepatocytes, decrease of glycogen deposits and fibrosis in periportal area were observed in HFD group. Liver MDA level was increased and GSH level was decreased in HFD group. Exercise treatment ameliorated these morphological and oxidative changes in HFD induced liver damage. CONCLUSION: Based on morphological and biochemical analysis, we could conclude that swimming training ameliorated obesity-induced liver damage by regulating lipid accumulation and oxidative damage.

Preclinical Assessment of Efficacy and Safety Analysis of CAR-T Cells (ISIKOK-19) Targeting CD19-Expressing B-Cells for the First Turkish Academic Clinical Trial with Relapsed/Refractory ALL and NHL Patients

Objective: Relapsed and refractory CD19-positive B-cell acute lymphoblastic leukemia (ALL) and non-Hodgkin lymphoma (NHL) are the focus of studies on hematological cancers. Treatment of these malignancies has undergone recent transformation with the development of new gene therapy and molecular biology techniques, which are safer and well-tolerated therapeutic approaches. The CD19 antigen is the most studied therapeutic target in these hematological cancers. This study reports the results of clinical-grade production, quality control, and in vivo efficacy processes of ISIKOK-19 cells as the first academic clinical trial of CAR-T cells targeting CD19-expressing B cells in relapsed/refractory ALL and NHL patients in Turkey. Materials and Methods: We used a lentiviral vector encoding the CD19 antigen-specific antibody head (FMC63) conjugated with the CD8-CD28-CD3ζ sequence as a chimeric antigen receptor (CAR) along with a truncated form of EGFR (EGFRt) on human T-lymphocytes (CAR-T). We preclinically assessed the efficacy and safety of the manufactured CAR-T cells, namely ISIKOK-19, from both healthy donors' and ALL/NHL patients' peripheral blood mononuclear cells. Results: We showed significant enhancement of CAR lentivirus transduction efficacy in T-cells using BX-795, an inhibitor of the signaling molecule TBK1/IKKƐ, in order to cut the cost of CAR-T cell production. In addition, ISIKOK-19 cells demonstrated a significantly high level of cytotoxicity specifically against a CD19+ B-lymphocyte cancer model, RAJI cells, in NOD/SCID mice. Conclusion: This is the first report of preclinical assessment of efficacy and safety analysis of CAR-T cells (ISIKOK-19) targeting CD19-expressing B cells in relapsed/refractory ALL and NHL patients in Turkey.

SARS-CoV-2 isolation and propagation from Turkish COVID-19 patients.

The novel coronavirus pneumonia, which was named later as coronavirus disease 2019 (COVID-19), is caused by the severe acute respiratory syndrome coronavirus 2, namely SARS-CoV-2. It is a positive-strand RNA virus that is the seventh coronavirus known to infect humans. The COVID-19 outbreak presents enormous challenges for global health behind the pandemic outbreak. The first diagnosed patient in Turkey has been reported by the Republic of Turkey Ministry of Health on March 11, 2020. In May, over 150,000 cases in Turkey, and 5.5 million cases around the world have been declared. Due to the urgent need for a vaccine and antiviral drug, isolation of the virus is crucial. Here, we report 1 of the first isolation and characterization studies of SARS-CoV-2 from nasopharyngeal and oropharyngeal specimens of diagnosed patients in Turkey. This study provides an isolation and replication methodology,and cell culture tropism of the virus that will be available to the research communities.

Protective effects of exercise on heart and aorta in high-fat diet-induced obese rats.

We investigated the protective effects of swimming exercise on high-fat diet-induced heart and aorta damage by evaluating oxidative stress and the endothelial nitric oxide (NO) system. Sprague Dawley rats were fed either standard chow (STD, 6% fat) or high-fat diet (HFD; 45% fat) for 18 weeks, with half of the animals trained by daily swimming sessions (EXC; 1 h per day for 5 days/week) for the last 6 weeks of the experimental period and half kept sedentary (SED). Heart and aorta tissues were prepared for routine light and electron microscopy evaluation. Endothelial NOS (eNOS) and inducible NOS (iNOS) distribution in the tissue samples were examined by immunohistochemistry. Biochemical examinations, including blood serum lipid profiles, malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), and tissue NO levels were measured. Deteriorated heart and aorta morphology, increased MDA levels and iNOS-immunoreactivity (iNOS-ir), as well as decreased GSH, NO, SOD, and eNOS-ir parameters were observed in the HFD + SED group. These morphological and biochemical parameters were ameliorated in the HFD + EXC group. Our study revealed that obesity-induced iNOS activation and increased oxidative stress in cardiac and aorta tissues. Exercise protected the obesity-induced cardiac and aortic tissue damage by modulating oxidant/antioxidant balance via involvement of the NO system.