Cholesteryl ester (CE)-rich lipid droplets (LDs) accumulate in steroidogenic tissues under physiological conditions and constitute an important source of cholesterol as the precursor for the synthesis of all steroid hormones. The mechanisms specifically involved in CE-rich LD formation have not been directly studied and are assumed by most to occur in a fashion analogous to triacylglycerol-rich LDs. Seipin is an endoplasmic reticulum protein that forms oligomeric complexes at endoplasmic reticulum-LD contact sites, and seipin deficiency results in severe alterations in LD maturation and morphology as seen in Berardinelli-Seip congenital lipodystrophy type 2. While seipin is critical for triacylglycerol-rich LD formation, no studies have directly addressed whether seipin is important for CE-rich LD biogenesis. To address this issue, mice with deficient expression of seipin specifically in adrenal, testis, and ovary, steroidogenic tissues that accumulate CE-rich LDs under normal physiological conditions, were generated. We found that the steroidogenic-specific seipin-deficient mice displayed a marked reduction in LD and CE accumulation in the adrenals, demonstrating the pivotal role of seipin in CE-rich LD accumulation/formation. Moreover, the reduction in CE-rich LDs was associated with significant defects in adrenal and gonadal steroid hormone production that could not be completely reversed by addition of exogenous lipoprotein cholesterol. We conclude that seipin has a heretofore unappreciated role in intracellular cholesterol trafficking.

In rural areas where the availability of utility grids is not possible, it is very difficult to install electric grid in those areas. The availability of plentiful sunlight is prone in those areas so the one of the best available fuel is solar energy or the electricity. In those areas solar cells are one of the most promising sources for the water supply and other agriculture use in the field. The intention of making the smart irrigation system using photo volcanic panels to provide efficient and easily available electric energy in the rural areas and agricultural fields. A humidity sensor is also installed in the dual axis solar tracker and mainly used for the purpose to sense the water requirement in the field according to the need of the soil. Pump is automatically activated when the humidity sensor senses the requirement of water in the field. This solar PV system based project is called Dual axis based solar tracker system. With the intention to monitor the movement by the sun, an LDR sensor is used which changes the direction of the PV panels according to the direction of sun. Since, it is a dual axis it can move horizontally as well as vertically Arduino microcontroller is used to make this solar tracker, it captures 40% more energy as compared to single and stationary mode solar tracker axis.

TRIB1 is a GWAS locus associated with plasma cholesterol and triglycerides (TG) levels. In mice, liver-specific overexpression of TRIB1 lowers plasma lipid levels. Berberine (BBR) is a natural lipid lowering drug that reduces plasma LDL-cholesterol (LDL-C), total cholesterol (TC) and TG in hyperlipidemic patients and in mice by mechanisms involving upregulation of hepatic LDL receptor (LDLR). Here, we demonstrated that BBR treatment reduced plasma LDL-C, TC and TG in LDLR wildtype (WT) mice fed a high fat and high cholesterol diet and it only lowered TG in LDLR WT mice fed a normal chow diet. In hypercholesterolemic LDLR deficient mice (Ldlr-/-), BBR treatment reduced plasma TG levels by 51% compared to the vehicle control without affecting plasma cholesterol levels. Hepatic gene expression analysis revealed that Trib1 mRNA levels were significantly elevated by BBR treatment in all three mouse models and increases of Trib1 mRNA expression were associated with reduced expression of lipogenic genes including Cebpa, Acc1 and Scd1. In vitro studies further demonstrate that BBR induces TRIB1 mRNA expression by a transcriptional mechanism via ERK signaling pathway. These new findings warrant future in vivo studies to determine the causal role of Trib1 in BBR-mediated TG lowering independent of LDLR regulation.

Objective— The objective of this study was to determine whether and how activation of farnesoid X receptor (FXR) by obeticholic acid (OCA), a clinical FXR agonist, modulates liver low-density lipoprotein receptor (LDLR) expression under normolipidemic conditions. Approach and Results— Administration of OCA to chow-fed mice increased mRNA and protein levels of LDLR in the liver without affecting the sterol-regulatory element binding protein pathway. Profiling of known LDLR mRNA-binding proteins demonstrated that OCA treatment did not affect expressions of mRNA degradation factors hnRNPD (heterogeneous nuclear ribonucleoprotein D) or ZFP36L1 but increased the expression of Hu antigen R (HuR) an mRNA-stabilizing factor. Furthermore, inducing effects of OCA on LDLR and HuR expression were ablated in Fxr −/− mice. To confirm the post-transcriptional mechanism, we used transgenic mice (albumin-luciferase-untranslated region) that express a human LDLR mRNA 3′ untranslated region luciferase reporter gene in the liver. OCA treatment led to significant rises in hepatic bioluminescence signals, Luc-untranslated region chimeric mRNA levels, and endogenous LDLR protein abundance, which were accompanied by elevations of hepatic HuR mRNA and protein levels in OCA-treated transgenic mice. In vitro studies conducted in human primary hepatocytes and HepG2 cells demonstrated that FXR activation by OCA and other agonists elicited the same inducing effect on LDLR expression as in the liver of normolipidemic mice. Furthermore, depletion of HuR in HepG2 cells by short interfering RNA transfection abolished the inducing effect of OCA on LDLR expression. Conclusions— Our study is the first to demonstrate that FXR activation increases LDLR expression in liver tissue by a post-transcriptional regulatory mechanism involving LDLR mRNA-stabilizing factor HuR.

Introduction: Curcumin is a naturally occurring polyphenol compound derived from the plant Curcuma longa. It is known for its pleiotropic effects including anti-inflammatory, antioxidant, anti-carcinogenic and antimicrobial properties. Several Studies have examined the role of this compound in inducing and maintaining remission in patients with inflammatory Bowel disease specially ulcerative colitis, but the outcome remain mixed. Specific limitations have been identified as low aqueous solubility and very short half-life for curcumin delivered in vivo. Thus, improving the biological half-life and bioavailability of curcumin to the inflamed mucosal tissue can significantly improve its clinical efficacy. To overcome these limitations, our collaborating laboratory generated chemically modified Curcumin - Eudragit-s100copolymer which significantly improved its water solubility and increased bioavailability, specifically to the colon. This compound was named as Ora-Curcumin-S. Methods: To test the clinical efficacy of the Ora-Curcumin-S in limiting inflammation-associated injury and improving the mucosal healing, we employed widely used mouse model of epithelial-injury induced colitis using the chemical Dextran Sodium Sulfate (DSS). Mice (C57/B16 strain, 6-8 weeks old) were exposed to DSS (2.5% w/v in drinking water) with or without oral administration (gavage) Ora-Curcumin-S for seven consecutive days. Colonoscopy was performed to evaluate longitudinal changes in mucosal health and mice were sacrificed on day 7 post DSS-administration. Endoscopic evaluation, % changes in body weight and the overall pathological scoring of the mucosal injury formed the basis of overall outcome Results: In vitro and in vivo analysis demonstrated that the aqueous solubility of Ora-Curcumin was significantly increased as well as its bio-availability in the colon. Colonoscopy evaluation at day-5 post DSS-administration demonstrated markedly less inflammation and injury in mice receiving DSS+Ora-Curcumin versus DSS alone. Furthermore, body weight analysis at the time of sacrifice demonstrated significantly less body weight loss in mice receiving DSS+Ora-Curcumin versus DSS alone (87.27±0.98 versus 91.77±0.84; P < 0.01). Similar beneficial effects of Ora-Curcumin administration was documented upon colon thickness (0.026±0.002 versus 0.034±0.002; P < 0.05) spleen weight (0.062±0.003 versus 0.083±0.006; P < 0.05) and the overall injury score 10.50±2.10 versus 25.00±4.12; p < 0.001) compared to mice receiving DSS alone. Conclusion: Cumulative outcome from our study provide strong evidence for potential use of Ora-Curcumin -S for therapeutic purposes. Our study not only re-affirms the known anti-inflammatory potential of Curcumin but also demonstrates how its chemical modification by nanoprecipitation can help improve its bioavailability in the colonic tissue, for potential use in IBD therapy and clinical management.

The present investigation was carried out at Main Experiment Station, Horticulture, Narendra Deva University of Agriculture and Technology, Kumarganj Faizabad (U.P.) under sodic soil condition during the years 2014-15 and 2015-16 to evaluate the response of organic manure, inorganic fertilizer and bio-fertilizer on physic-chemical characters of bael fruit cv. Narendra Bael-9. The experiment was laid out with treatments viz. T1-100% NPK, T2-50 Kg FYM, T3-50 Kg FYM + 100% NPK, T4-50 Kg FYM + 75% NPK, T5-50 Kg FYM + 50% NPK, T6-50 Kg FYM + 200g each (Azotobacter + PSB), T7-50 Kg FYM + 100% NPK + 200g each (Azotobacter + PSB), T8-50 Kg FYM + 75% NPK + 200g each (Azotobacter + PSB) and T9-50 Kg FYM + 50% NPK + 200g each (Azotobacter + PSB) and replicated four times with Randomized Block Design. The physico-chemical characters were found significantly superior over all other treatments while physical characters like specific gravity, number of cavity/fruit and shell thickness (mm) were found non-significant during both the years of experimentation (2014-15 and 2015-16) respectively.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) impedes low‑density lipoprotein (LDL) receptor (LDLR)-mediated LDL-cholesterol uptake and has hence emerged as a critical regulator of serum cholesterol levels and a new therapeutic target for the treatment of hypercholesterolemia. Statins have been shown to elevate circulating PCSK9 levels by stimulating PCSK9 gene transcription, which reduces the clinical efficacy of statin in LDL‑cholesterol reduction. The transcription of PCSK9 is partially controlled by the hepatocyte nuclear factor 1 (HNF1) binding site embedded in the proximal region of its promoter. In this study, we utilized adenoviral shRNA delivery vectors to generate liver-specific knockdown of HNF1α (Ad‑shHNF1α) or HNF1β (Ad‑shHNF1β) in hamsters to examine the impact of reduced hepatic expression of HNF1 transcription factors on statin‑induced elevation of PCSK9 expression and serum cholesterol levels. We showed that the administration of rosuvastatin (RSV) to normolipidemic hamsters significantly augmented hepatic PCSK9 expression and serum PCSK9 levels. In addition, RSV treatment increased hepatic HNF1α protein levels without a clear effect on HNF1α mRNA expression. Injection of Ad-shHNF1α or Ad‑shHNF1β into hamsters both blunted RSV‑induced elevation of PCSK9 serum concentration and hepatic mRNA and protein levels, which led to significant increases in liver LDLR protein abundance. Furthermore, hepatic depletion of HNF1 factors lowered circulating total cholesterol and non‑high density lipoprotein cholesterol levels in RSV‑treated hamsters. Our study demonstrates that both HNF1α and HNF1β are positive regulators of hepatic PCSK9 transcription in hamster species and that transient, liver-specific knockdown of either HNF1α or HNF1β could antagonize the RSV‑induced elevation of serum PCSK9 and reduce circulating cholesterol levels.

The farnesoid X receptor (FXR) plays critical roles in plasma cholesterol metabolism, in particular HDL-cholesterol (HDL-C) homeostasis. Obeticholic acid (OCA) is a FXR agonist being developed for treating various chronic liver diseases. Previous studies reported inconsistent effects of OCA on regulating plasma cholesterol levels in different animal models and in different patient populations. The mechanisms underlying its divergent effects have not yet been thoroughly investigated. The scavenger receptor class B type I (SR-BI) is a FXR-modulated gene and the major receptor for HDL-C. We investigated the effects of OCA on hepatic SR-BI expression and correlated such effects with plasma HDL-C levels and hepatic cholesterol efflux in hyperlipidemic hamsters. We demonstrated that OCA induced a time-dependent reduction in serum HDL-C levels after 14 days of treatment, which was accompanied by a significant reduction of liver cholesterol content and increases in fecal cholesterol in OCA-treated hamsters. Importantly, hepatic SR-BI mRNA and protein levels in hamsters were increased to 1.9- and 1.8-fold of control by OCA treatment. Further investigations in normolipidemic hamsters did not reveal OCA-induced changes in serum HDL-C levels or hepatic SR-BI expression. We conclude that OCA reduces plasma HDL-C levels and promotes transhepatic cholesterol efflux in hyperlipidemic hamsters via a mechanism involving upregulation of hepatic SR-BI.

Long-chain acyl-CoA synthetase 1 (ACSL1) plays a key role in fatty acid metabolism. To identify novel transcriptional modulators of ACSL1, we examined ACSL1 expression in liver tissues of hamsters fed a normal diet, a high fat diet, or a high cholesterol and high fat diet (HCHFD). Feeding hamsters HCHFD markedly reduced hepatic Acsl1 mRNA and protein levels as well as acyl-CoA synthetase activity. Decreases in Acsl1 expression strongly correlated with reductions in hepatic Srebp2 mRNA level and mature Srebp2 protein abundance. Conversely, administration of rosuvastatin (RSV) to hamsters increased hepatic Acsl1 expression. These new findings were reproduced in mice treated with RSV or fed the HCHFD. Furthermore, the RSV induction of acyl-CoA activity in mouse liver resulted in increases in plasma and hepatic cholesterol ester concentrations and reductions in free cholesterol amounts. Investigations on different ACSL1 transcript variants in HepG2 cells revealed that the mRNA expression of C-ACSL1 was specifically regulated by the sterol regulatory element (SRE)-binding protein (SREBP) pathway, and RSV treatment increased the C-ACSL1 abundance from a minor mRNA species to an abundant transcript. We analyzed 5'-flanking sequence of exon 1C of the human ACSL1 gene and identified one putative SRE site. By performing a promoter activity assay and DNA binding assays, we firmly demonstrated the key role of this SRE motif in SREBP2-mediated activation of C-ACSL1 gene transcription. Finally, we demonstrated that knockdown of endogenous SREBP2 in HepG2 cells lowered ACSL1 mRNA and protein levels. Altogether, this work discovered an unprecedented link between ACSL1 and SREBP2 via the specific regulation of the C-ACSL1 transcript.

Peroxisome proliferator-activated receptor delta (PPARD) is a transcription factor regulating many important genes involved in lipid and glucose metabolism. Hepatic lysophosphatidylcholine acyltransferase 3 (LPCAT3) plays critical functions in TG secretion and ER stress response due to its unique ability to catalyze the incorporation of polyunsaturated fatty acids into phospholipids. Previous studies identified liver X receptor as the transcription factor controlling LPCAT3 expression in liver tissue. Here we showed for the first time that the hepatic LPCAT3 gene is transcriptionally regulated by PPARD in vitro and in vivo . Treatments of HepG2, Huh-7, and Hepa 1-6 cells with PPARD specific agonists (L-165041 and GW0742) increased LPCAT3 mRNA levels 2-3 fold of control. Adenovirus-mediated knockdown of PPARD in HepG2 cells reduced the basal level of LPCAT3 mRNA by approximately 40% compared to control and further attenuated the induction of PPARD agonist on LPCAT3 gene expression. To demonstrate the direct regulation of PPARD on LPCAT3 gene transcription, we analyzed human LPCAT3 promoter sequence and identified two putative PPAR responsive elements (PPRE-1, -171; PPRE-2, -727) within 1 kb of the proximal LPCAT3 promoter. LPCAT3 promoter assays demonstrated strong inductions of the reporter activity by PPARD agonists. Mutational analysis of the human LPCAT3 promoter revealed that the PPRE-1 is involved in conferring responsiveness to PPARD agonist mediated transcriptional activity. Direct binding of PPARD to PPRE-1 sequence was demonstrated by chromatin immunoprecipitation and gel retardation assays. Finally, we provided in vivo evidence showing that activation of PPARD by L-165041 in mice increased LPCAT3 mRNA abundance in the liver. Altogether, these new findings identify LPCAT3 as a direct PPARD target gene and suggest a novel function of PPARD in regulation of phospholipid metabolism through LPCAT3.

There are several modes are available for teaching to the farmers through distance training system, out of which telecast of video programme, broadcast of audio programmes, telecounselling etc. are eco-friendly modes, solving the farmers problems.The telecounselling is a ecofriendly and easy system of distance training, which is available in rural area in the form of mobiles and landline.The experiment on telecounselling for improvement of potato yield was tried in the villages Bhawanipur and Daipur of district Kannauj and Rajpura, Baghauli, Jasharau and Shaidpur of district Mainpuri for the first time.The soil of experimental site was sandy loam with poor fertility.There is certain drawback in the cultivation of potato.The farmers are not using the recommended agronomic practices, resulting in, the low yield obtained from potato.The farm families of potato growers were linked with C.S.Azad University of Agriculture and Technology, Kanpur and KVK, Kannauj.The growers put up their problems of potato cultivation before the scientists through telecouselling mode of distance training as and when required.The scientists solved their problems immediately with same mode of distance training.The cultivation of potato was recommended with three varieties i.e.Kufri Bahar, Kufri Pukhraj and Kufri Pushkar through telecounselling mode.Cultivars Kufri Bahar, Kufri Pukhraj and Kufri Pushkar gave tuber yield by 331.55 q/ha, 354.22 q/ha and 383.71 q/ha, respectively.The growth and yield traits were concordant to yield obtained from potato cultivars.The highest net return Rs. 123569/ha and BCR 1:2.15 were obtained from adoption of cultivation of cultivar Kufri Pushkar closely followed by the net return Rs. 106031/ha and BCR 1:2.00 available from the cultivation of Kufri Pukhraj.

The transcription factors hepatic nuclear factor (HNF)1α and HNF1β can bind to the HNF1 site on the proprotein convertase subtilisin/kexin type 9 (PCSK9) promoter to activate transcription in HepG2 cells. However, it is unknown whether one or both HNF1 factors are obligatory for transactivating hepatic PCSK9 gene expression in vivo. We developed shRNA adenoviral constructs (Ad-shHNF1α and Ad-shHNF1β) to examine the effects of knockdown of HNF1α or HNF1β on PCSK9 expression and its consequent impact on LDL receptor (LDLR) protein levels in cultured hepatic cells and liver tissue. We demonstrated that infection with Ad-shHNF1α, but not Ad-shHNF1β, markedly reduced PCSK9 mRNA expression in HepG2 cells with a concomitant increase in LDLR protein abundance. Injecting Ad-shHNF1α in mice fed a normal diet significantly (∼50%) reduced liver mRNA expression and serum concentration of PCSK9 with a concomitant increase (∼1.9-fold) in hepatic LDLR protein abundance. Furthermore, we observed a modest but significant reduction in circulating LDL cholesterol after knockdown of HNF1α in these normolipidemic mice. Consistent with the observation that knockdown of HNF1β did not affect PCSK9 mRNA or protein expression in cultured hepatic cells, Ad-shHNF1β infection in mice resulted in no change in the hepatic mRNA expression or serum content of PCSK9. Altogether, our study demonstrates that HNF1α, but not HNF1β, is the primary positive regulator of PCSK9 transcription in mouse liver.

Background: High fructose diet (HFD) induces dyslipidemia and insulin resistance in experimental animals and humans with incomplete mechanistic understanding. By utilizing mice and hamsters as in vivo models, we investigated whether high fructose consumption affects serum PCSK9 and liver LDL receptor (LDLR) protein levels.Results: Feeding mice with an HFD increased serum cholesterol and reduced serum PCSK9 levels as compared with the mice fed a normal chow diet (NCD). In contrast to the inverse relationship in mice, serum PCSK9 and cholesterol levels were co-elevated in HFD-fed hamsters. Liver tissue analysis revealed that PCSK9 mRNA and protein levels were both reduced in mice and hamsters by HFD feeding, however, liver LDLR protein levels were markedly reduced by HFD in hamsters but not in mice. We further showed that circulating PCSK9 clearance rates were significantly lower in hamsters fed an HFD as compared with the hamsters fed NCD, providing additional evidence for the reduced hepatic LDLR function by HFD consumption. The majority of PCSK9 in hamster serum was detected as a 53 kDa N-terminus cleaved protein. By conducting in vitro studies, we demonstrate that this 53 kDa truncated hamster PCSK9 is functionally active in promoting hepatic LDLR degradation.Conclusion: Our studies for the first time demonstrate that high fructose consumption increases serum PCSK9 concentrations and reduces liver LDLR protein levels in hyperlipidemic hamsters. The positive correlation between circulating cholesterol and PCSK9 and the reduction of liver LDLR protein in HFD-fed hamsters suggest that hamster is a better animal model than mouse to study the modulation of PCSK9/LDLR pathway by atherogenic diets. Published by Elsevier Ireland Ltd.

The hepatic expression of low-density lipoprotein (LDL) receptor (LDLR) gene is regulated primarily at the transcriptional level by a sterol-regulatory element (SRE) in its proximal promoter region which is the site of action of SRE-binding protein 2 (SREBP2). However whether additional cis-regulatory elements contribute to LDLR transcription has not been fully explored. We investigated the function of a putative peroxisome proliferator-activated receptor (PPAR)-response element (PPRE) sequence motif located at -768 to -752 bases upstream of the transcription start site of human LDLR gene in response to PPARδ activation. Promoter luciferase reporter analyses showed that treating HepG2 cells with PPARδ agonist L165041 markedly increased the activity of a full-length LDLR promoter construct (pLDLR-1192) without any effects on the shorter promoter reporter pLDLR-234 that contains only the core regulatory elements SRE-1 and SP1 sites. Importantly, mutation of the PPRE sequence greatly attenuated the induction of the full-length LDLR promoter activity by L165041 without affecting rosuvastatin (RSV)-mediated transactivation. EMSA and ChIP assay further confirmed the binding of PPARδ to the LDLR-PPRE site. Treating HepG2 cells with L165041 elevated the mRNA and protein expressions of LDLR without affecting the LDLR mRNA decay rate. The induction of LDLR expression by PPARδ agonist was further observed in liver tissue of mice and hamsters treated with L165041. Altogether, our studies identify a novel PPRE-mediated regulatory mechanism for LDLR transcription and suggest that combined treatment of statin with PPARδ agonists may have advantageous effects on LDLR expression.

BACKGROUND: In central India (M.P.) there is high prevalence of hepatitis; of which hepatitis B contributes as a major cause of hepatitis. In our area there were no major efforts to contain the disease. According to one estimate most of the hepatitis B occurs due to negligence transfusion of infected blood (Hepatitis B). Therefore we have tried to found percentage of population infected with this disease. MATERIAL AND METHODS: Study design - Prospective study. For screening donors we have used one step rapid test for the detection of HBsAg and confirmatory test was done by ELISA method. STATISTICAL ANALYSIS: Statistical analysis was done by using Minitab 17 Pennsylvania, USA. RESULTS: In our study we found 2.8% HBsAg infected blood among healthy donors; which was significantly (P < 0.05) more in voluntary donors in comparison to replacement donors. Male between age groups 39-48 were found highly infected in comparison with other age groups. We also found prevalence was highest in Hindu community (2.9%) and more in working group (3.9%) in comparison to students and farmers. CONCLUSION: For effective control of transfusion of infected blood and to prevent its spread, we have to improve community knowledge towards vaccination; educational programs for blood bank personals; and government should open blood banks at Community Health Center level.

Background: CETP inhibitors block the transfer of cholesteryl ester from HDL-C to VLDL-C and LDL-C, thereby raising HDL-C and lowering LDL-C. In this study, we explored the effect of CETP inhibitors on hepatic LDL receptor (LDLR) and PCSK9 expression and further elucidated the underlying regulatory mechanism.Results: We first examined the effect of anacetrapib (ANA) and dalcetrapib (DAL) on LDLR and PCSK9 expression in hepatic cells in vitro. ANA exhibited a dose-dependent inhibition on both LDLR and PCSK9 expression in CETP-positive HepG2 cells and human primary hepatocytes as well as CETP-negative mouse primary hepatocytes (MPH). Moreover, the induction of LDLR protein expression by rosuvastatin in MPH was blunted by cotreatment with ANA. In both HepG2 and MPH ANA treatment reduced the amount of mature form of SREBP2 (SREBP2-M). In vivo, oral administration of ANA to dyslipidemic C57BL/6J mice at a daily dose of 50 mg/kg for 1 week elevated serum total cholesterol by approximately 24.5% (p < 0.05%) and VLDL-C by 70% (p < 0.05%) with concomitant reductions of serum PCSK9 and liver LDLR/SREBP2-M protein. Finally, we examined the in vitro effect of two other strong CETP inhibitors evacetrapib and torcetrapib on LDLR/PCSK9 expression and observed a similar inhibitory effect as ANA in a concentration range of 1-10 mu M.Conclusion: Our study revealed an unexpected off-target effect of CETP inhibitors that reduce the mature form of SREBP2, leading to attenuated transcription of hepatic LDLR and PCSK9. This negative regulation of SREBP pathway by ANA manifested in mice where CETP activity was absent and affected serum cholesterol metabolism. Published by Elsevier Ireland Ltd.