Gross and histopathological features of tuberculosis in cattle, buffalo and spotted deer (Axis axis) caused by Mycobacterium orygis.

Mycobacterium orygis has been isolated from several cases of tuberculosis in various species of animal in India but documentation of the histopathological lesions caused by this organism is scant. Lung and liver tissues with caseous nodules from cattle (n = 8), lung samples from spotted deer (Axis axis) (n = 5) and lung and mediastinal lymph node samples from buffalo (n = 9) were subjected to histopathology and isolation of Mycobacterium spp. Isolation was carried out using the BACTEC MGIT 960 Automated Mycobacterial Detection System and acid-fast positive cultures were identified to species level using polymerase chain reaction (PCR) employing published primer pairs. Three M. orygis isolates (two from cattle, one from deer) were obtained, whole genome sequenced and the sequences submitted to the National Center for Biotechnology Information Sequence Read Archive. Eight samples (four cattle, one deer and three buffalo) were confirmed as M. orygis positive by PCR. Histopathological examination of the M. orygis-PCR-positive cattle samples revealed acid-fast organisms in lung sections along with macrophages, epithelioid cells, lymphocytes and Langhans giant cells. Granuloma stages I to IV were seen in the cattle and buffalo samples and stage III in the spotted deer sample. This report is the first description of the gross and histopathological lesions of tuberculosis caused by M. orygis in buffalo and documents the gross and histopathological findings of M. orygis tuberculosis in cattle and deer.

Modulation of PPAR-γ, SREBP-1c and inflammatory mediators by luteolin ameliorates ß-cell dysfunction and renal damage in a rat model of type-2 diabetes mellitus.

BACKGROUND: Natural products have been recommended as a complementary therapy for type 2 diabetes mellitus (T2DM) due to constraints of safety and tolerability of existing anti-diabetic agents. Luteolin exhibits anti-diabetic and anti-inflammatory effects. Hence, the impact of luteolin on glucose homoeostasis and organ damage was investigated in high-fat diet (HFD) and streptozotocin (STZ) induced T2DM in rats. METHODS AND RESULTS: Male Wistar rats were maintained on HFD (provided 55% energy as fat) for 10 days. Subsequently, a single dose of 40 mg/kg STZ was injected intraperitoneally on the 11th day. Seventy-two hours after STZ administration, diabetic rats with established hyperglycemia (fasting serum glucose > 200 mg/dL) were randomized into different groups having six rats each and orally administered either 0.5% hydroxy propyl cellulose or pioglitazone (10 mg/kg) or luteolin (50 mg/kg or 100 mg/kg) once daily for 28 days, while continuing HFD for respective groups. Luteolin significantly reduced hyperglycaemia, homoeostasis model assessment (HOMA) of insulin resistance (HOMA-IR) levels, and improved hypoinsulinemia and HOMA of b-cell function (HOMA-B) in a dose-dependent manner. Increased TNF-α, IL-6 and NFκB levels in diabetic rats were significantly regulated. Additionally, luteolin significantly augmented PPAR-γ expression while attenuating sterol regulatory element binding protein-1c (SREBP-1c) expression. Histopathological scrutiny validated that luteolin effectively attenuated HFD-STZ-induced injury in pancreatic ß-cells and kidneys to near normalcy. CONCLUSION: Our study showed that luteolin ameliorated hyperglycemia and improved hypoinsulinemia, ß-cell dysfunction, and renal impairment in HFD-STZ-induced diabetic rats by attenuating inflammation and dysregulated cytokine secretion through modulation of PPAR-γ, TNF-α, IL-6 and NF-kB expression and down-regulation of SREBP-1c.

Expression of iron regulatory proteins in full-term swine placenta.

In swine, even though the pregnant sows were with iron abundance, the inborn iron reserve of piglets was compromised. This indicates the insufficiency of molecular machinery involved in local placental iron flux. Here, we investigated the expression of iron regulatory proteins like hepcidin and ferroportin and also their association with iron reserve, inflammation and oxidative stress in placenta of full-term pregnant sows (n = 6). Amplification and sequencing of placental DNA confirmed the presence of hepcidin (MN579557) and ferroportin (MN565887) sequences and their 100% identity with existing GenBank data. Real-time amplification of placental mRNA revealed significant higher expression of hepcidin (p < .05) than ferroportin. Western blot analysis of placental tissues revealed specific bands for both hepcidin (~8 kDa) and ferroportin (~62 kDa) molecules. Immunohistochemistry revealed the immunoreactivity for both proteins in the cytoplasm and membrane of trophoblastic cells of the placenta. Hepcidin and ferroportin expressions were positively associated with placental non-haem iron reserve (p < .0001; p = .033), lipid peroxidation (p = .0060; p < .0001) and reactive oxygen species level (p = .0092; p = .0292). Hepcidin expression was positively associated with interleukin - 6 (p = .0002) and interferon gamma (p < .0001) expressions but ferroportin expression was negatively associated with interleukin-6 (p = .0005), interleukin-1ß (p = .0226) and interferon gamma (p = .0059) expressions. This indicates hepcidin and ferroportin may have a role in controlling the local placental iron flux by acting as a molecular bridge between iron trafficking and inflammation.

Lipolytic and antiadipogenic effects of (3,3-dimethylallyl) halfordinol on 3T3-L1 adipocytes and high fat and fructose diet induced obese C57/BL6J mice.

Aegle marmelos Correa., (Rutaceae) is a medium sized tree distributed in South East Asia and used traditionally for the management of obestiy and diabetes. In this study the lipolytic and antiadipogenic effects of (3,3-dimethylallyl) halfordinol (Hfn) isolated from leaves of A. marmelos have been investigated. Intracellular lipid accumulation was measured by oil red O staining and glycerol secretion. The expression of genes related to adipocyte differentiation was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). Hfn decreased intracellular triglyceride accumulation and increased glycerol release in a dose dependent manner (5-20 µg/ml) in differentiated 3T3-L1 adipocytes. In high fat diet fed C57/BL 6J mice, treatment with Hfn for four weeks reduced plasma glucose, insulin and triglyceride levels and showed a significant reduction in total adipose tissue mass by 37.85% and visceral adipose tissue mass by 62.99% at 50mg/kg b.w. concentration. RT-PCR analyses indicated that Hfn decreased the expression of peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT enhancer binding protein α (CEBPα) and increased the expression of sterol regulatory enzyme binding protein (SREBP-1c), peroxisome proliferator-activated receptor α (PPARα), Adiponectin and Glucose transporter protein 4 (GLUT4) compared to the high fat diet group. These results suggested that Hfn decreased adipocyte differentiation and stimulated lipolysis of adipocytes. This study justifies the folklore medicinal uses and claims about the therapeutic values of this plant for the management of insulin resistance and obesity.

Hepatoprotective role of Abelmoschus esculentus (Linn.) Moench., on carbon tetrachloride-induced liver injury.

CONTEXT: Chronic liver disease has become a global health problem. The research for prominent herbal agents for the management of liver diseases is widely increased. OBJECTIVE: The root of Abelmoschus esculentus (Linn.) Moench., (Malvaceae) has been used as a remedy for liver disorders. The aim of the present study was to evaluate the antioxidant and hepatoprotective effects of the ethanol extract of A. esculentus root. MATERIALS AND METHOD: The antioxidant effect was assessed using DPPH and hydroxy radical scavenging assays. The hepatoprotective effect of the extract was evaluated using CCl4 intoxicated HepG2 cell line and Wistar rats by estimating the levels of hepatic and antioxidant markers. RESULTS: The extract of A. esculentus showed IC50 values of 270.99 and 532.86 µg/mL for DPPH and hydroxy radical scavenging assays, respectively. The incubation of HepG2 cells with CCl4 drastically decreased the cell viability and increased the leakage of transaminases. Pre-treatment with the extract significantly restored the cell death by 31.25 and 39.04% at 200 and 400 µg/mL concentrations, respectively. The reduction of ALT leakage by the treatment was 18.62, 38.59 and 52.15% compared to the CCl4 treated cells at 100, 200 and 400 µg/mL, respectively. In in-vivo experiments also the treatment reduced the levels of transaminases, ALP, MDA, total bilirubin and hepatic TNFα levels as well as increased the antioxidant levels in a dose dependent manner. Histological observations of liver sections showed reduction in steatosis, necrosis and inflammation. CONCLUSION: The results substantiated the hepatoprotective activity of A. esculentus through its antioxidant capacity.