Loss-of-function W4645R mutation in the RyR2-caffeine binding site: implications for synchrony and arrhythmogenesis.

AIMS: Previous studies have identified RyR2 W4645R mutation, located in the caffeine-binding site, to associate with CPVT1 pathology. Caffeine binding to its site is thought to displace the carboxyl-terminal domain to Ca2+-binding, allowing the tryptophan residue (W4645) to regulate Ca2+ sensitivity of RyR2. To gain insights into regulation of RyR2 Ca2+-binding and its interaction with caffeine-binding site, we introduced W4645R-RyR2 point mutation via CRISPR/Cas9 gene-editing in human induced pluripotent stem cell-derived cardiomyocytes (hiPSCCMs) and characterized their Ca2+-signaling phenotype compared to WT hiPSCCMs. METHODS AND RESULTS: W4645R-RyR2 cardiomyocytes had: (1) no significant change in ICa magnitude or voltage-dependence; (2) slightly reduced CICR; (3) altered relaxation kinetics of Ca2+-transients with no change in isoproterenol sensitivity; (4) complete loss of caffeine-triggered Ca2+ release; (5) larger SR Ca2+ leak resulting in 40 % lower SR Ca2+ content, as determined by myocytes' response to 4-CmC; (6) lower incidence of calcium sparks and asynchronous spontaneous SR Ca2+ releases. CONCLUSIONS: W4645R-RyR2 mutation induces loss of caffeine-triggered SR Ca2+ release and enhances SR Ca2+ leak that underlie asynchronous spontaneous Ca2+ releases, triggering arrhythmia and impairing cardiac function.

Assessing the impact of dental restorative materials on fibroblast cells: an immunohistochemical and ELISA analysis.

In this study, our aim was to investigate the effects of restorative materials such as composite, compomer and high viscosity glass ionomer, which are frequently used in dentistry, on L929 fibroblast cells by evaluating the oxidative stress parameters, pro- and anti-inflammatory cytokines, and apoptosis markers. L929 fibroblast cells were cultured, and dental filling materials were applied in two doses (50 and 100 µl). Immunohistochemical staining was performed for experimental groups with Anti-Bax and Anti-Caspase 9 antibodies. Then, ELISA technique was used to detect the level of TNF-alpha, TGF-beta, IL-1-beta, IL-6, IL-10, LPO and CAT. In the light of the data, the examined dental filling materials were effective on increasing the TGF-beta, IL-10, LPO and CAT levels, and decreasing the TNF-alpha, IL-1-beta, and IL-6 levels. The histological micrographs were also support the issues. When the levels of H-score in Caspase 9 labeled micrographs were evaluated, the mean of the control group was lower than the mean of the experimental groups. Biocompatibility varies according to the content of the material, the amount of residual monomer, and its solubility. Although all the experimental groups have cytotoxic effects, the least effect is seen in the Omnichroma group.

Protective effects of a natural product, paeoniflorin, on ischemia reperfusion injury on rat ovary tissue: histopathological, immunohistochemical, and biochemical study.

In this study, the main hypothesis is that paeoniflorin may inhibit some cellular processes such as oxidative stress and inflammation. For this reason, we aimed to investigate the potential protective effects of a natural compound, paeoniflorin, on rat model of ovarian ischemia-reperfusion injury by detecting the oxidative stress parameters and inflammatory process parameters. 42 female Wistar-albino rats were divided into 6 random groups. The rats were subjected to 3-hour ischemia and 3-hour reperfusion process. Then, paeoniflorin at doses of 25, 50 and 100 mg/kg were applied 30 min before the reperfusion. The levels of pro-inflammatory (IL-1-ß, IL-6, TNF-α) and anti-inflammatory (IL-10, TGF-ß) cytokines were measured by ELISA. Similarly, IL-6, IL-10, TNF-α, NF-κB p65) positivity rates were detected by immunohistochemical staining. Additionally, oxidative stress parameters (MDA, GSH, SOD) were measured by tissue biochemistry. Ischemia-reperfusion injury caused significant increase in the levels of SOD, MDA, TNF-α, IL-1-ß, IL-6 and NF-κB p65, while paeoniflorin treatments improved the related parameters in a dose-dependent manner. As a conclusion, our findings support the evidence that paeoniflorin has a potential protective effects on ovarian ischemia-reperfusion injury. Further detailed studies should be performed to shed light the molecular mechanism of these protective effects.

Morphological and Functional Analysis of Cardiac Ameliorations in Elderly Rats Supplemented with a Magnolol Extract Complex / Análisis Morfológico y Funcional de las Mejoras Cardíacas en Ratas Ancianas Suplementadas con un Complejo de Extracto de Magnolol

SUMMARY: Magnolia bark extract supplementation has an anti-oxidative role in mammalians. However, its role in physiological aged-associated heart insufficiency is not known yet. Therefore, we investigated the effects of a magnolia bark complex, including magnolol and honokiol components (MAHOC), in elderly rat hearts (24-month-old aged group). One group of aged rats was supplemented with MAHOC (400 mg/kg/d, for 12 weeks) besides the standard rat diet while the second group of elderly rats and adult rats (to 6-month- old adult-group) were only fed with the standard rat diet. The morphological analysis using light microscopy has shown marked myofibrillar losses, densely localized fibroblasts, vacuolizations, infiltrated cell accumulations, and collagen fibers in the myocardium of the elderly rats compared to the adults. We also detected a markedly increased amount of degenerated cardiomyocytes including the euchromatic nucleus. The MAHOC supplementation of the elderly rats provided marked ameliorations in these abnormal morphological changes in the heart tissue. Furthermore, electrophysiological analysis of electrocardiograms (ECGs) in the supplemented group showed significant attenuations in the prolonged durations of P-waves, QRS-complexes, QT-intervals, and low heart rates compared to the unsupplemented elderly group. The biochemical analysis also showed significant attenuations in the activity of arylesterase and total antioxidant status in the myocardium of the supplemented group. We further determined significant attenuations in the activity of a mitochondrial enzyme succinate dehydrogenase, known as a source of reactive oxygen species (ROS), and the decreased level of ATP/ADP in the heart homogenates of the supplemented group. Moreover, under in vitro conditions by using an aging-mimicked cardiac cell line induced by D-galactose, we demonstrated that MAHOC treatment could provide prevention of depolarization in mitochondria membrane potential and high-level ROS production. Overall, our data presented significant myocardial ameliorations in physiological aging-associated morphological alterations parallel to the function and biochemical attenuations with MAHOC supplementation, at most, through recoveries in mitochondria.

La suplementación con extracto de corteza de magnolia tiene un papel antioxidante en los mamíferos, sin embargo, su rol en la insuficiencia cardíaca asociada al envejecimiento fisiológico aún no se conoce. Por lo anterior, investigamos los efectos de un complejo de corteza de magnolia, incluidos los componentes magnolol y honokiol (MAHOC), en corazones de ratas seniles (grupo de edad de 24 meses). La alimentación de grupo de ratas seniles se complementó con MAHOC (400 mg/kg/d, durante 12 semanas) además de la dieta estándar, mientras que el segundo grupo de ratas seniles y ratas adultas (hasta el grupo de adultos de 6 meses) solo recibió la dieta estándar para ratas. El análisis morfológico mediante microscopía óptica ha mostrado marcadas pérdidas miofibrilares, fibroblastos densamente localizados, vacuolizaciones, acumulaciones de células infiltradas y fibras de colágeno en el miocardio de las ratas seniles en comparación con las adultas. También detectamos una cantidad notablemente mayor de cardiomiocitos degradados, incluido el núcleo eucromático. La suplementación con MAHOC de las ratas seniles proporcionó mejoras marcadas en estos cambios morfológicos anormales en el tejido cardiaco. Por otra parte, el análisis de los electrocardiogramas (ECG) en el grupo suplementado mostró atenuaciones significativas en las duraciones prolongadas de las ondas P, los complejos QRS, los intervalos QT y las frecuencias cardíacas bajas, en comparación con el grupo de ratas seniles sin suplementación alimenticia. El análisis bioquímico también mostró atenuaciones significativas en la actividad de la arilesterasa y el estado antioxidante total en el miocardio del grupo suplementado. Determinamos además atenuaciones significativas en la actividad de la enzima mitocondrial succinato deshidrogenasa, conocida como fuente de especies reactivas de oxígeno (ROS), y la disminución del nivel de ATP/ADP en los homogeneizados de corazón del grupo suplementado. Además, en condiciones in vitro mediante el uso de una línea de células cardíacas, imitando el envejecimiento inducido por D- galactosa, demostramos que el tratamiento con MAHOC podría prevenir la despolarización en el potencial de membrana de las mitocondrias y la producción de ROS de alto nivel. En general, nuestros datos presentaron mejoras miocárdicas significativas en alteraciones morfológicas asociadas con el envejecimiento fisiológico paralelas a la función y atenuaciones bioquímicas con la suplementación con MAHOC, como máximo, a través de recuperaciones en las mitocondrias.

The effects of cisplatin and jaceosidin on SH-SY5Y neuroblastoma cells: an electron microscopic, molecular and biochemical study.

In this study, our aim was to show both the single and combined effects of cisplatin and jaceosidin in SHSY-5Y neuroblastoma cells. For this purpose, we used MTT cellular viability assay, Enzyme-Linked Immunosorbent Assay (ELISA), Transmission Electron Microscopy (TEM), Immunofluorescence Staining Assay (IFA) and Western blotting (WB) assay. According to MTT findings, IC50 dose was detected as 50 µM cisplatin and 160 µM jaceosidin co-application. Therefore, experimental groups were finally selected as control, cisplatin, 160 µM jaceosidin and Cisplatin +160 µM jaceosidin. Cell viability was decreased in all groups, and the IFA findings confirmed the viability analysis. WB data indicated that matrix metalloproteinase 2 and 9 levels, as indicators of metastasis, decreased. While LPO and CAT levels increased in all treatment groups, it was observed that the activity of SOD decreased. When TEM micrographs were investigated, cellular damages were determined. In the light of these results, it can be said that cisplatin and jaceosidin have a potential to increase the effects of each other synergistically.

Two flavonoids, baicalein and naringin, are effective as anti-inflammatory and anti-oxidant agents in a rat model of polymicrobial sepsis.

INTRODUCTION: In this study, our aim was to investigate the possible protective and therapeutic effects of these two flavonoids, baicalein, and naringin, in 50 and 100 mg/kg doses applied both before and after cecal ligation and puncture (CLP) procedures in a polymicrobial sepsis rat model, and evaluate the possible contribution of oxidative and inflammatory markers by immunological, biochemical, molecular, and histopathological methods. METHODS: Sixty-six Wistar albino rats were divided into 11 groups. The pro-inflammatory (TNF-alpha, IL-1-beta, and IL-6) and anti-inflammatory (TGF-beta and IL-10) cytokine levels were measured by ELISA technique. CD3, CD68, and nuclear factor kappa B positivity rates were detected by immunohistochemical methods. Oxidative stress parameters (MDA, SOD, and GSH) were measured by tissue biochemistry. RESULTS: Sepsis caused a significant increase in all pro-inflammatory cytokine levels and MDA activity. Also, it led to an increase in the positivities of CD3, CD68, and NF-κB markers. However, especially pre-CLP doses of baicalein and naringin inhibited the inflammation process by suppressing pro-inflammatory and increasing anti-inflammatory cytokine levels, as well as regulating the oxidative stress process by normalizing the oxidant/anti-oxidant enzyme levels. CONCLUSION: Both pre- and post-application of baicalein and naringin are quite effective to prevent sepsis-caused cellular processes. This protective and therapeutic effects by baicalein and naringin in animals with sepsis seems to be originated from the high antioxidant capacity and inhibition of pro-inflammatory cytokine production. Thus, those natural agents may prove to be valuable protective agent against septic shock.

Alterations in hippocampal neurogenesis and hippocampal insulin signaling pathway in rat with metabolic syndrome.

Objectives: We aimed to examine the level of hippocampal neurogenesis, and assess learning and anxiety and the level of some proteins involving insulin signaling pathways in rats with Metabolic Syndrome (MetS); and to reveal the relationship among them. Materials and Methods: Totally, 30 Wistar-albino rats were used. The rats were divided into three groups: Control, MetS, and MetS+Ins. Immunohistochemical staining was performed to evaluate the levels of neurogenesis markers; Doublecortin (DCX), Neuronal-Differentiation-1 (NeuroD1), Ki67, and Neuronal nuclear protein (NeuN). Then, cleaved caspase-3 and TUNEL labeling were performed to detect the level of apoptosis. Additionally, behavior tests were performed to evaluate the learning-memory levels and anxiety-like behaviors. Insulin, Insulin Receptor (IR), Insulin Receptor Substrate (IRS2), glucose transporter (GLUT)-3, and GLUT4 protein expression levels were analyzed to evaluate the possible changes in the insulin signaling pathway. Results: An increase in anxiety with memory deficiency was observed in MetS. In the hippocampus of MetS, an increase was detected in the level of apoptosis, whereas a decrease was detected in the expression level of the neurogenesis marker. Insulin secretion and IR levels decreased in hippocampal neurons. We observed that GLUT3 and GLUT4 levels increased because of the non-activated insulin signaling pathway. Conclusion: We think that the insulin signaling pathway may have an effect on the decreased neurogenesis in the MetS group. So, the evaluation of the Mitogen-activated protein kinase (MAPK) pathway and the investigation of the effect of endoplasmic reticulum stress on this pathway will be among the targets of our future studies.

The effects of dragon fruit (Hylocereus polyrhizus) extract on indomethacin-induced stomach ulcer in rats.

The aim of this study was to investigate the effects of red dragon fruit (Hylocereus polyrhizus) extract (DFE) on the stomach in ulcer model induced by indomethacin in rats. Effects of DFE were evaluated in indomethacin-induced gastric damage model on Sprague-Dawley rats. Experimental model: all rats were fasted for 24 h. At the end of this period, DFE was administered to the ulcer-induced groups. One hour after this application, a dose of 25 mg/kg of indomethacin was applied by oral gavage to all groups except the HEALTHY and DFE1000 groups. Six hours after indomethacin administration, the rats were euthanized with high-dose anesthesia and the experiment was terminated. Macroscopic and microscopic analyses for investigating ulcerative area, molecular and biochemical analyses for oxidative damages investigation and molecular analyses for the effect mechanism of indomethacin and DFE were conducted on stomach tissues in the study. While oxidative stress-associated markers such as MDA, BAX, and Caspase 3 increased dramatically in the indomethacin group, GSH antioxidant levels decreased. It was observed that these parameters were significantly improved in DFE 500 mg/kg and DFE 1000 mg/kg groups compared to ulcer group, and the results of especially DFE 1000 mg/kg group were similar to famotidine group. We observed that our histopathological findings also supported all our other findings. Dragon fruit extract was protected against indomethacin-induced ulcer damage by decreased MDA levels, increased GSH levels, and inhibition of Caspase 3, BAX, and Cox-2, and activation of Cox-1. PRACTICAL APPLICATIONS: People of all ages around the world suffer from gastric ulcer which is one of the most common gastrointestinal ailments. The etiological factors of the disease are using of cigarette and alcohol, nutritional deficiencies, infections, and using non-steroidal anti-inflammatory drugs which use frequent and indiscriminate. Indomethacin is one of the NSAIDs and is commonly preferred to induce ulcer modeling in rats due to its gastric toxicity potential. Current anti-ulcer drugs have many serious side effects. Patients who suffered from gastric ulcer tend to discontinue the drug because of side effects. Therefore, patients need new agents that are non-toxic, have few side effects, and are easily accessible anti-ulcer drugs. Dragon fruit, as a medicinal herb, is highly valuable and widely used in traditional medicine, and may provide gastroprotective activity. Studies have shown that H. polyrhizus has antioxidant activities. We consider the effects of dragon fruit extract (DFE) to be a therapeutic drug for an indomethacin-induced ulcer model.

In Vivo and In Vitro Cardioprotective Effect of Gossypin Against Isoproterenol-Induced Myocardial Infarction Injury.

The aim of the study was to examine the protective effects and possible mechanism of gossypin against isoproterenol (ISO)-mediated myocardial damage in vivo and H9c2 cell damage in vitro. H9c2 cells were categorized into five groups. Viability was evaluated with MTT and LDH release in H9c2 cells. Apoptotic parameter analysis was performed with cytochrome c (Cyt-c), caspase-3 (CASP-3), and BCL2/Bax mRNA expression levels. In vivo, gossypin was administered orally to mice at doses of 5, 10, and 20 mg/kg for 7 days. ISO groups were injected with isoproterenol (150 mg/kg) subcutaneously (on 8th and 9th) for 2 days. Afterward, lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB) levels and Troponin-I (Tn-I) amount from their serum, oxidative stress parameters superoxide dismutase (SOD) activity, glutathione (GSH) and malondialdehyde (MDA) levels, and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1ß (IL-1 ß), and NF-kB mRNA expression levels with inflammatory markers from heart tissue were evaluated. In addition, IL-1B, BCL-2, and cas-3 immunohistochemical staining was performed from heart tissue and TNF-a level was measured by ELISA method. Administration of Gossypin protected the cells by dose-dependent, eliminating the reduced cell viability and increased LDH release of ISO in H9c2 cells. In mice serum analyses, increased LDH, CK-MB levels, and Tn-I levels were normalized by gossypin. ISO administration in heart tissue is regulated by gossypin with increased SOD activity, GSH amount, TNF-α, IL-6, IL-1ß, and NF-kB mRNA expression levels and decreased MDA amount. Overall, the present results demonstrated that gossypin has a potential cardioprotective treatment for ischemic heart disease on in vivo and in vitro.

Effects of aerobic exercise on lipopolysaccharide-induced experimental acute lung injury in the animal model of type 1 diabetes mellitus.

NEW FINDINGS: What is the central question of this study? We evaluated the effects of diabetes and exercise on lipopolysaccharide-induced acute lung injury. By providing a comprehensive analysis of redox status, blood gases and histological parameters, we aimed to contribute to the ongoing debate in the literature. What are the main findings and its importance? We demonstrated the preventive effect of exercise, but diabetes did not alter the severity of acute lung injury. ABSTRACT: Acute lung injury (ALI) is a life-threatening respiratory condition. Diabetes (DM) is a metabolic disease characterized by hyperglycaemia. There is an ongoing debate concerning whether there is a protective effect of diabetes in ALI. Exercise is a special type of physical activity that has numerous beneficial effects. The aim of our study was to investigate the effects of diabetes and exercise on the prognosis of ALI. Male Wistar albino rats were divided into two groups (sedentary and exercise). Both groups were divided into four subgroups: Control, ALI, DM, DM+ALI (n = 6 each). Diabetes was induced by injection of streptozotocin (50 mg/kg i.p.). The maximal exercise capacity was determined with the incremental load test. Animals were exercised on a treadmill for 45 min at 70% of maximal exercise capacity, 5 days a week for 12 weeks. Acute lung injury was induced by intratracheal injection of lipopolysaccharide (100 µg/100 g body weight) 24 h before the end of the experiment. We performed arterial blood gas analysis. Redox status was measured in both plasma and lung tissue. Malondialdehyde and 8-hydroxy-2'-deoxyguanosine levels were measured in lung tissue. Lung tissue was evaluated histologically. Acute lung injury caused significant damage in the lung tissue, which was verified histologically, with an increase in oxidative stress parameters. Exercise prevented the lung damage induced by ALI and reduced oxidative stress in the lung tissue. Diabetes did not alter the magnitude of damage done by ALI. Exercise showed a protective effect against DM and ALI in rats. The effect of DM was insignificant for the prognosis of ALI.

Histologic and Immunohistochemical Assessment of the Effect of Various Biomaterials on New Bone Formation in the Maxillary Sinus Floor Augmentation Procedure.

PURPOSE: To assess the effect of different kinds of biomaterials placed with maxillary sinus floor augmentation (MSFA) on bone regeneration. MATERIALS AND METHODS: Thirty-six New Zealand rabbits were used in the study. A standardized method of surgical approach was used for MSFA under anesthesia in all groups. The procedure was performed for each animal. Six separate groups with 12 cases were created. In group 1, no graft was used in MSFA. Advanced platelet-rich fibrin (A-PRF), absorbable collagen cone (ACC), venous blood, the combination of ACC and platelet-rich plasma (PRP), and the combination of ACC and enamel matrix derivative (EMD) were used in groups 2, 3, 4, 5, and 6, respectively. At the end of 4 and 12 weeks, three rabbits from each group were sacrificed by applying high-dose anesthetic, and samples were examined histologically and immunohistochemically. RESULTS: Groups 2 and 5 showed significantly increased new bone formation compared with groups 1 and 4, 4 weeks after MSFA (P < .05). Twelve weeks after sinus floor augmentation, groups 2, 3, 5, and 6 showed significantly higher new bone formation than group 1 (P < .05). Groups 2 and 5 showed significantly higher hard tissue response than groups 1 and 4 at the end of 4 weeks (P < .05). Groups 5 and 6 demonstrated significantly higher hard tissue response than group 1 at the end of 12 weeks (P < .05). Group 5 also showed significantly higher hard tissue response than group 4 at the end of 12 weeks (P < .05). Immunohistochemical analysis showed a significant difference between the osteocalcin scores of groups 2 and 4 and group 1 at the end of 4 and 12 weeks (P < .05). There was also a statistically significant difference between osteocalcin scores at 4 and 12 weeks for groups 1 and 5 (P < .05). When osteopontin scores were compared, there was no significant difference between groups at 4 and 12 weeks (P > .05). Groups 1, 2, and 5 showed significant changes in osteopontin scores between 4 and 12 weeks (P < .05). CONCLUSION: The combination of ACC and PRP and the combination of ACC and EMD showed increased new bone formation and hard tissue response. A-PRF also showed promising results in new bone formation at the end of both 4 and 12 weeks. The usage of ACC as a carrier for liquid-form biomaterials may be more beneficial than the usage of ACC alone.

What is the restorative effect of VEGF inhibitor bevacuzimab against subarachnoid hemorrhage in an experimental model?

BACKGROUND: This study investigated the effect of vascular endothelial growth factor (VEGF) inhibitor bevacuzimab (BVZ) on the rabbit basilar artery using an experimental subarachnoid hemorrhage (SAH) model. METHODS: Eighteen adult male New-Zealand white rabbits were randomly divided into three groups: a control group (n = 6), SAH group (n = 6), and SAH+BVZ group (n = 6). Experimental SAH was created by injecting autologous arterial blood into the cisterna magna. In the treatment group, the subjects were administered a daily dose of 10 mg/kg, intravenous BVZ for 2 days after the SAH. Basilar artery diameters were measured with magnetic resonance angiography (MRA) 72 h after the SAH in all groups. After 72 h, whole brains, including the upper cervical region, were obtained from all the animals after perfusion and fixation of the animal. The wall thickness, luminal area, and the apoptosis at the basilar arteries were evaluated in all groups. RESULTS: BVZ significantly prevented SAH-induced vasospasm confirmed in vivo with MRA imaging with additional suppression of apoptosis on basilar artery wall. DISCUSSION: VEGF inhibition with BVZ has shown to have a vasospasm and apoptosis attenuating effect on basilar artery in a SAH model.

Effects of eccentric exercise on different slopes.

OBJECTIVES: Eccentric contraction occurs when the muscle lengthens under tension. Damage-induced responses seen in the muscle after eccentric exercise usually experienced by sedentary individuals. This study aims to investigate muscle damage on different slopes. METHODS: 32 male Wistar albino rats randomly divided into four groups: sedentary, horizontal running, and eccentric exercise (-8°, -16°) groups. Animals ran for 90 min with the speed of 25 m/s for five days. After 48h from the last exercise, rats were sacrificed, and plasma creatine kinase (CK), heat shock protein 70 (HSP70) levels were examined. Plasma and soleus total oxidant/antioxidant status (TOS-TAS) and histological changes of soleus muscle assessed. RESULTS: CK and HSP70 significantly increased in 16° EE group. TOS increased at 16° EE and 8° EE, but oxidative stress index (OSI) was only high at 8° EE group. Mononuclear cell infiltration and the angiogenesis increased in soleus after eccentric exercise, and there was a correlation with slope. Sarcomere breaks were detected in 16° EE group also in a correlation with slope. CONCLUSIONS: Consequently, sedentary individuals are vulnerable to injuries induced by eccentric contraction. Therefore, our study provides information for reconsidering rehabilitation and training programs.

Montelukast attenuates radioactive I131-induced pulmonary damage on rats.

PURPOSE: Radioactive I131 (RAI) therapy is a standard method to ablate the remnant thyroidal tissue after total thyroidectomy and its metastases in differentiated thyroid carcinomas; however, I131 also accumulates in nonthyroidal tissue, which may cause adverse effects and limit the I131 dose required for treatment. We hypothesized that montelukast, a known agent with anti-inflammatory and anti-oxidant properties, would ameliorate the radiation-induced histopathological characteristics such as pneumonitis and fibrosis in rat lungs after RAI. METHODS: Fifty female Wistar albino rats were randomly separated into five groups of 10. Group 1 was the control group; Group 2 was administered RAI only; Group 3 was administered RAI and montelukast, Group 4 was administered RAI after total thyroidectomy and Group 5 was administered RAI and montelukast after total thyroidectomy. All rats were sacrificed after 12 weeks and the lungs were evaluated in the histological examination to determine the degree of inflammation and fibrosis and for immunohistochemical (IHC) staining for tissue expression of IL-1, IL-6 and TNF-alpha and TGF-beta. RESULTS: The RAI-administered groups, Group 2 and Group 4, were significantly different from the control group, however, the groups medicated with both RAI and montelukast, Group 3 and Group 5, were not significantly different from the control group. All histopathological and IHC parameters were significantly less in the groups administered with montelukast compared to the groups not administered with montelukast. CONCLUSIONS: The results of this study demonstrated the radioprotective effect of montelukast in the pulmonary system through histopathological and IHC examination.

Histopathologic Analysis of Tamoxifen on Epidural Fibrosis.

BACKGROUND: Epidural fibrosis is a challenging topic in spinal surgery. This phenomenon constitutes one of the main reasons behind postlaminectomy syndrome or failed back surgery syndrome, which leads to persistent back and leg pain in association with compression and/or stretching the nerve root or the dura. The exact mechanism of action in epidural fibrosis is complex and remains uncertain. Excessive deposition of collagen, fibronectin, and dermatan sulfate, known as the "extracellular matrix," and decrease of tissue cellularity results in epidural fibrosis. The most investigated and important actor in epidural fibrosis as well as in other forms of aberrant wound healing is presumed to be transforming growth factor-1ß formation. Tamoxifen (TAM), a synthetic nonsteroidal antiestrogen used in breast cancer, is also effective in inhibiting fibroblast proliferation via downregulation of transforming growth factor-1ß. METHODS: Twenty-four adult male rats were randomly divided into 3 groups. Laminectomy was the sole intervention in the control group. Spongostan was placed in the operation lodge after laminectomy in the second group. In the treatment group, TAM was administrated orally after laminectomy. Epidural fibrosis, dural thickness, inflammatory response, and arachnoidal involvement were evaluated and graded histopathologically. RESULTS: Epidural fibrosis, dural thickness, and inflammatory response in the subjects treated with TAM were significantly less than in the control and Spongostan group and the differences were statistically significant. Although arachnoidal involvement was observed in a subject in the TAM group, the differences between all groups weren't statistically significant. CONCLUSIONS: Tamoxifen reduced epidural fibrosis, dural thickness, and inflammatory response after laminectomy in rats.

Punica granatum L. Juice Attenuates Experimental Cerebral Vasospasm in the Rabbit Subarachnoid Hemorrhage Model: A Basilar Artery Morphometric Study and Apoptosis.

Background This study investigated the effect of Punica granatum L. (pomegranate) juice on the rabbit basilar artery in an experimental subarachnoid hemorrhage (SAH) model. Methods Eighteen adult male New Zealand white rabbits were randomly divided into three groups: a control group (n = 6), SAH group (n = 6), and SAH + treatment group (n = 6). Basilar artery diameter was measured with magnetic resonance angiography (MRA) in all groups at the beginning of the study. Experimental SAH was created by injecting autologous arterial blood into the cisterna magna. In the treatment group, the subjects were administered a daily dose of 30 ml/kg pomegranate juice via gastric gavage for 4 days after the SAH. The SAH group and SAH + treatment group underwent cerebral MRA after 72 hours. After a neurologic score assessment, all the animals were killed. The wall thickness and lumen area of the basilar artery were measured histometrically in all groups, and the apoptotic cell percentage in the artery was identified. The mean diameter of the basilar artery during MRA was measured. Results Pomegranate improved neurologic functions compared with the SAH group (p < 0.01). The mean basilar artery diameter on MRA in the SAH + treatment group was larger than in the SAH group and smaller than in the control group (p < 0.01 and p < 0.05, respectively). The mean vessel wall thickness value in the SAH + treatment group was lower than in the SAH group (p < 0.01), whereas there was no difference between the control and the SAH + treatment group (p > 0.05). The apoptotic cell rate in the SAH + treatment group was significantly lower than in the SAH group (p < 0.001). Evaluation of the basilar artery luminal area showed no difference between the three groups (p > 0.05). Discussion Pomegranate was shown to have a vasospasm- attenuating effect on the basilar artery in the rabbit SAH model for the first time in our study.

Development of an LC-MS/MS analytical method for the simultaneous measurement of aldehydes from polyunsaturated fatty acids degradation in animal feed.

Knowing that polyunsaturated fatty acids can lead to the formation of potentially toxic aldehydes as secondary oxidation products, an analytical method using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) detection was developed to measure the concentration of eight aldehydes in animal feed: malondialdehyde (MDA), 4-hydroxy-2-nonenal (4-HNE), 4-hydroxy-2-hexenal (4-HHE), crotonaldehyde (CRT), benzaldehyde (BNZ), hexanal (HXL), 2,4-nonadienal, and 2,4-decadienal. The developed method was validated according to the criteria and procedure described in international standards. The evaluated parameters were specificity/selectivity, recovery, precision, accuracy, uncertainty, limits of detection and quantification, using the concept of accuracy profiles. These parameters were determined during experiments conducted over three different days with ground Kellogg's® Corn Flakes® cereals as model matrix for animal feed and spiked at different levels of concentration. Malondialdehyde, 4-HHE, 4-HNE, crotonaldehyde, benzaldehyde, and hexanal can be analyzed in the same run in animal feed with a very good accuracy, with recovery rates ranging from 86 to 109% for a working range going from 0.16 to 12.50 mg/kg. The analysis of 2,4-nonadienal and 2,4-decadienal can also be performed but in a limited range of concentration and with a limited degree of accuracy. Their recovery rates ranged between 54 and 114% and coefficient of variation for the intermediate precision between 11 and 25% for these two compounds. Copyright © 2016 John Wiley & Sons, Ltd.

Incremento de la expresión de Bax (proapoptótico) y disminución de la expresión de Bcl-2 (antiapoptótico) en recién nacidos con enterocolitis necrosante / Increase in pro-apoptotic Bax expression and decrease in anti-apoptotic Bcl-2 expression in newborns with necrotizing enterocolitis

Antecedentes/Objetivo: El objetivo de este estudio fue determinar si se producía un incremento de la expresión de Bax (proapoptótico) y una disminución de la expresión de Blc-2A1 (antiapoptótico) en el intestino de los recién nacidos con enterocolitis necrosante. Materiales y métodos: Comparamos a ocho pacientes recién nacidos de manera consecutiva sometidos a resección intestinal debido a enterocolitis necrosante con ocho recién nacidos sometidos a resección intestinal debido a atresia ileal. La evaluación histopatológica de la lesión tisular y la apoptosis se realizó mediante microscopía óptica y el método TUNEL. El nivel de ARNm en los genes apoptóticos (CASP3, CASP6, CASP7, Bax, BIRC2) y antiapoptóticos se evaluó con el método de matriz de RCP (PCR array). La expresión de proteínas se evaluó mediante inmunohistoquímica. Resultados: Los puntajes de las lesiones tisulares y los puntajes medios de apoptosis fueron significativamente más altos en el grupo con enterocolitis necrosante en comparación con el grupo de referencia (p < 0,01). La expresión de los genes proapoptóticos aumentó significativamente en el grupo con enterocolitis necrosante frente al grupo de referencia (p < 0,01). La expresión del gen Bcl-2A1 (antiapoptótico) disminuyó significativamente en el grupo con enterocolitis necrosante (p < 0,01). La expresión de las proteínas Bax y CASP3 aumentó significativamente en el grupo con enterocolitis necrosante (p < 0,01). Conclusión: Según nuestros datos, la alteración del equilibrio entre la expresión de Bax (proapoptótico) y la expresión de Bcl-2A1 (antiapoptótico) en el lugar de la lesión es un posible mecanismo de la patogenia en recién nacidos que presentan enterocolitis necrosante.

Background/Aim. The aim of the present study was to find out if there is an increase in the expression of pro-apoptotic Bax and reduction in expression of anti-apoptotic Blc-2A1 in newborn intestines with necrotizing enterocolitis (NEC). Material and Methods. We compared 8 consecutive newborn patients undergoing bowel resection for NEC with 8 neonates undergoing intestinal resection for ileal atresia. Histopathological evaluation of tissue injury and apoptosis was performed by using light microscopic examination and TUNEL method. The mRNA level of apoptotic (CASP3, CASP6, CASP7, Bax, BIRC2) and anti-apoptotic genes were evaluated by PCR array method. Protein expression was assessed by immunohistochemistry. Results. Tissue injury scores and mean apoptosis scores were significantly higher in NEC group when compared with control group (p <0.01). Expression of pro-apoptotic genes were significantly increased in NEC group when compared with control group (p <0.01). Expression of anti-apoptotic Bcl-2A1 gene was significantly decreased in NEC group, (p <0.01). Protein expression of Bax and CASP3 was significantly increased in NEC group, (p <0.01). Conclusion. Our data in humannewborns suggest that alteration of the balance between pro-apoptotic Bax expression and anti-apoptotic Bcl-2A1 expression in the site of injury is a possible mechanism in the pathogenesis of NEC.

Increase in pro-apoptotic Bax expression and decrease in anti-apoptotic Bcl-2 expression in newborns with necrotizing enterocolitis. / Incremento de la expresión de Bax (proapoptótico) y disminución de la expresión de Bcl-2 (antiapoptótico) en recién nacidos con enterocolitis necrosante.

BACKGROUND/AIM: The aim of the present study was to find out if there is an increase in the expression of pro-apoptotic Bax and reduction in expression of anti-apoptotic Blc-2A1 in newborn intestines with necrotizing enterocolitis (NEC). MATERIALS AND METHODS: We compared 8 consecutive newborn patients undergoing bowel resection for NEC with 8 neonates undergoing intestinal resection for ileal atresia. Histopathological evaluation of tissue injury and apoptosis was performed by using light microscopic examination and TUNEL method. The mRNA level of apoptotic (CASP3, CASP6, CASP7, Bax, BIRC2) and anti-apoptotic genes were evaluated by PCR array method. Protein expression was assessed by immunohistochemistry. RESULTS: We compared 8 consecutive newborn patients undergoing bowel resection for NEC with 8 neonates undergoing intestinal resection for ileal atresia. Histopathological evaluation of tissue injury and apoptosis was performed by using light microscopic examination and TUNEL method. The mRNA level of apoptotic (CASP3, CASP6, CASP7, Bax, BIRC2) and anti-apoptotic genes were evaluated by PCR array method. Protein expression was assessed by immunohistochemistry. CONCLUSIONS: Our data in humannewborns suggest that alteration of the balance between pro-apoptotic Bax expression and anti-apoptotic Bcl-2A1 expression in the site of injury is a possible mechanism in the pathogenesis of NEC.

ANTECEDENTES/OBJETIVO: El objetivo de este estudio fue determinar si se producía un incremento de la expresión de Bax (proapoptótico) y una disminución de la expresión de Blc-2A1 (antiapoptótico) en el intestino de los recién nacidos con enterocolitis necrosante. MATERIALES Y MÉTODOS: Comparamos a ocho pacientes recién nacidos de manera consecutiva sometidos a resección intestinal debido a enterocolitis necrosante con ocho recién nacidos sometidos a resección intestinal debido a atresia ileal. La evaluación histopatológica de la lesión tisular y la apoptosis se realizó mediante microscopía óptica y el método TUNEL. El nivel de ARNm en los genes apoptóticos (CASP3, CASP6, CASP7, Bax, BIRC2) y antiapoptóticos se evaluó con el método de matriz de RCP (PCR array). La expresión de proteínas se evaluó mediante inmunohistoquímica. RESULTADOS: Los puntajes de las lesiones tisulares y los puntajes medios de apoptosis fueron significativamente más altos en el grupo con enterocolitis necrosante en comparación con el grupo de referencia (p < 0,01). La expresión de los genes proapoptóticos aumentó significativamente en el grupo con enterocolitis necrosante frente al grupo de referencia (p < 0,01). La expresión del gen Bcl-2A1 (antiapoptótico) disminuyó significativamente en el grupo con enterocolitis necrosante (p < 0,01). La expresión de las proteínas Bax y CASP3 aumentó significativamente en el grupo con enterocolitis necrosante (p < 0,01). CONCLUSIONES: Según nuestros datos, la alteración del equilibrio entre la expresión de Bax (proapoptótico) y la expresión de Bcl-2A1 (antiapoptótico) en el lugar de la lesión es un posible mecanismo de la patogenia en recién nacidos que presentan enterocolitis necrosante.