Therapeutic enhancing potential of piracetam with diethylstilbestrol in prevention of grand-mal seizures in rats: inhibition of PI3K/Akt/mTOR signaling pathway and IL-1ß, IL-6, TNF-α cytokines levels.

OBJECTIVE: Epilepsy, a neurodegenerative disorder, continues to throw challenges in the therapeutic management. The current study sought to ascertain if the therapeutic interactions between piracetam and diethylstilbestrol may prevent grand-mal seizures in rats. MATERIALS AND METHODS: Piracetam (PIR; 10 and 20 mg/kg) and diethylstilbestrol (DES; 10 and 20 mg/kg) alone as a low-dose combination were administered to rats for 14 days. The electroshock (MES; 180 mA, 220 V for 0.20 s) was delivered via auricular electrodes on the last day of treatment and rats were monitored for convulsive behavior. To elucidate the mechanism, hippocampal mechanistic target of rapamycin (mTOR) and interleukin (IL)-1ß, IL-6 and tumor necrotic factor-alpha (TNF-α) levels were quantified. Hippocampal histopathology was conducted to study the neuroprotective effect of drug/s. In vitro studies and in silico studies were conducted in parallel. RESULTS: To our surprise, the low dose of the combination regimen of PIR (10 mg/kg) and DES (10 mg/kg) unfolded synergistic anti-seizure potential, with brimming neuroprotective properties. The mechanism could be related to a significant reduction in the levels of hippocampal mTOR and proinflammatory cytokines. The docking scores revealed higher affinities for phosphatidylinositol 3-kinase (PI3K) in co-bound complex, and when docking DES first, while better affinities for protein kinase B (Akt) were revealed when docking PIR first (both drugs bind cooperatively as well). This indicated that the entire PI3K/Akt/mTOR signaling pathway is intercepted by the said combination. In addition, the % of cell viability of HEK-293 cells [pre-exposed to pentylenetetrazol (PTZ)] was increased by 327.29% compared to PTZ-treated cells (toxic control; 85.16%). CONCLUSIONS: We are the first to report the promising efficacy of the combination (PIR 10 mg/kg + DES 10 mg/kg) to restrain seizures and epileptogenic changes induced by electroshock by a novel mechanism involving inhibiting the PI3K/Akt/mTOR signaling.

Evaluation of metabolic and oxidative profile in ovine pregnancy toxemia and to determine their association with diagnosis and prognosis of disease.

This study was conducted on ewes with pregnancy toxemia (PT) with an attempt to evaluate metabolic and oxidative profile in subclinical and clinical ovine pregnancy toxemia and to determine their association with diagnosis and prognosis of the disease. A total of 20 ewes having beta-hydroxy butyric acid (ß-HBA) > 2.5 mmol/L and proven clinical sings of PT, categorized as clinical PT (CPT); 12 ewes having ß-HBA 0.8-2.5 mmol/L and no clinical signs of PT, categorized at subclinical PT (SPT); and 10 ewes having ß-HBA ≤ 0.8 mmol/L, categorized as healthy control (CON) were enrolled. Among 20 CPT ewes, 11 had negative outcomes (non-survivors), six ewes had positive outcomes (survivors), and three were lost during follow-up. A significant increase in non-esterified fatty acid, ß-HBA, triglycerides, gamma-glutamyl transferase, lactate dehydrogenase, and malondialdehyde levels and a significant decrease in fructosamine were observed in CPT and SPT compared to CON. A significant increase in cholesterol, aspartate amino transferase, and creatinine kinase and a significant decrease in albumin, potassium, calcium, superoxide dismutase, and catalase were observed in CPT only. Glucose was significantly decreased in SPT only. The highest area under the curve (AUC) was observed for fructosamine (89.7% and 87.5% for CPT and SPT, respectively) with the optimum cutoff point calculated on the basis of maximum sensitivity (SE) and specificity (SP) being 0.607 mmol/L (SE: 89.3% and SP: 72.2%) and 1.005 mmol/L (SE: 90.0% and SP: 75.3%) for CPT and SPT, respectively. At the cutoff limit of 0.607 mmol/L and 1.005 mmol/L, the odds ratio was 10.8 and 8.0 for CPT and SPT, respectively. A significant decrease in fructosamine and potassium and a significant increase in creatinine, lactate dehydrogenase, and malondialdehyde were observed in non-survivors compared to survivors. It was thus concluded that fructosamine was the best diagnostic indicator of both CPT and SPT followed by non-esterified fatty acid. Fructosamine, creatinine, potassium, lactate dehydrogenase, and malondialdehyde were the best prognostic indicators of PT.

Effect of physiological status and parity on metabolic and trace element profile of crossbred Rambouillet sheep of Himalayan region.

The study was designed to evaluate the effect of physiological status and parity on metabolic profile in crossbred Rambouillet ewes of the Himalayan region. The study was conducted on 20 ewes divided into two groups, primiparous (PP) and multiparous (MP), with 10 ewes in each group. Blood samples were collected on 4- and 1-week pre-lambing and 1- and 4-week post-lambing to measure metabolic parameters and minerals. The glucose (p < 0.01), total plasma protein (TPP) (p < 0.05), albumin (p < 0.05), blood urea nitrogen (BUN) (p < 0.05), cholesterol (p < 0.05), triglyceride (p < 0.01), high-density lipoprotein cholesterol (HDL-C) (p < 0.05), calcium (Ca) (p < 0.01), phosphorus (Pi) (p < 0.05), magnesium (Mg) (p < 0.01), copper (Cu) (p < 0.05), and zinc (Zn) (p < 0.01) levels revealed significant change along the time with the concentration decreasing from 3-week pre-lambing to immediate post-lambing; thereafter, levels increased steadily. Significant increase (p < 0.01) was observed in non-esterified fatty acid (NEFA), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), iron (Fe) (p < 0.05), and bilirubin (p < 0.05) concentrations along the sampling time. No group difference was observed in any of the parameters; however, parity and time interaction was observed in glucose, NEFA, GGT, Ca, and Pi. While NEFA levels were significantly high in pre-lambing in PP ewes compared to MP ewes, the post-lambing levels were significantly high in MP ewes. Pre-lambing levels of GGT were at par between the two groups; however, post-lambing levels were significantly high in MP ewes. Glucose, Ca, and Pi were low during pre-lambing in PP ewes and post-lambing in MP ewes. The result showed that ewes show a significant change in metabolic profile and trace minerals during late gestation and immediate postpartum; however, these changes were more pronounced during late gestation in primiparous and post-lambing in multiparous.

Sarcoptic mange in dogs: Its effect on liver, oxidative stress, trace minerals and vitamins.

The present study was aimed to determine the effect on liver, associated oxidative stress, trace element and vitamin alteration in dogs with sarcoptic mange. A total of 24 dogs with clinically established diagnosis of sarcoptic mange, divided into two groups, severely infested group (n=9) and mild/moderately infested group (n=15), according to the extent of skin lesions caused by sarcoptic mange and 6 dogs as control group were included in the present study. In comparison to healthy control hemoglobin, PCV, and TEC were significantly (P<0.05) decreased in dogs with sarcoptic mange however, significant increase in TLC along with neutrophilia and lymphopenia was observed only in severely infested dogs. The albumin, glucose and cholesterol were significantly (P<0.05) decreased and globulin, ALT, AST and bilirubin were significantly (P<0.05) increased in severely infested dogs when compared to other two groups. Malondialdehyde (MDA) levels were significantly (P<0.01) higher in dogs with sarcoptic mange, with levels highest in severely infested groups. Activity of superoxide dismutase (SOD) (P<0.05) and catalase were significantly (P<0.01) lower in sarcoptic infested dogs when compared with the healthy control group. Zinc and copper levels in dogs with sarcoptic mange were significantly (P<0.05) lower when compared with healthy control group with the levels lowest in severely infested group. Vitamin A and vitamin C levels were significantly (P<0.05) lower in sarcoptic infested dogs when compared to healthy control. From the present study, it was concluded that sarcoptic mange in dogs affects the liver and the infestation is associated with oxidant/anti-oxidant imbalance, significant alteration in trace elements and vitamins.

Evaluation of trace elements, oxidant/antioxidant status, vitamin C and ß-carotene in dogs with dermatophytosis.

The aim of the study was to determine zinc, copper and iron levels, erythrocyte oxidant/antioxidant status, vitamin C and ß-carotene in dogs with dermatophytosis. A total of 23 dogs with clinically established diagnosis of dermatophytosis by trichogram and positive fungal culture and six dogs as control were included in this study. On cultural examination 52.17% fungal isolates were found to be Microsporum canis, 30.43% were Trichophyton mentagrophytes and 17.39% were M. gypseum. In comparison to healthy control, the dogs with dermatophytosis had significantly lower levels of zinc (P < 0.01), copper (P < 0.05), ß-carotene and vitamin C levels (P < 0.05) and activities of superoxide dismutase (SOD) (P < 0.05) and catalase (P < 0.01), whereas the iron (P < 0.05) and malondialdehyde (MDA) (P < 0.01) levels were significantly increased. On correlation analysis, SOD activity was observed to be positively correlated (P < 0.05) with zinc and copper in both healthy and dermatophytosis affected dogs. In dermatophytosis affected dogs the MDA levels were negatively correlated (P < 0.05) with iron, ß-carotene levels and the activities of antioxidant enzymes; SOD and catalase. Our results demonstrated that dermatophytosis in dogs is associated with significant alteration in oxidant/antioxidant balance and trace elements. It might be secondary consequence of dermatophytosis infection or contributing factor in its pathogenesis.

Trace minerals status and antioxidative enzyme activity in dogs with generalized demodecosis.

The present study was aimed to determine the levels of trace elements zinc, copper, iron, erythrocyte oxidant/anti-oxidant balance, vitamin C and ß-carotene in dogs with generalized demodecosis. A total of 24 dogs with clinically established diagnosis of generalized demodecosis and 6 dogs as control were included in the study. In comparison to healthy control, zinc and copper levels were significantly (P<0.01) lower in dogs with generalized demodecosis, whereas iron levels were significantly (P<0.01) higher. Malondialdehyde (MDA) levels were significantly (P<0.01) higher in diseased dogs whereas activity of superoxide dismutase (SOD) and catalase were significantly (P<0.01) lower. ß-carotene and vitamin C levels were significantly (P<0.05) lower in diseased dogs when compared to healthy control. SOD activity was positively correlated with zinc (rs=0.65, rs=0.71 and P<0.05) and copper (rs=0.51, rs=0.63 and P<0.05) in both healthy and diseased dogs. MDA levels were negatively correlated with iron (rs=-0.49, rs=-0.78 and P<0.05), ß-carotene (rs=-0.26, P>0.05; rs=-0.54, P<0.05, respectively) in both healthy and diseased dogs and with SOD activity in diseased dogs only (rs=-0.68, P<0.05). From the present study, it was concluded that generalized demodecosis in dogs is associated with significant alteration in trace elements and oxidant/anti-oxidant imbalance and this imbalance might be secondary to changes caused by demodectic mange.