No abstract

mTORC1 is a protein kinase important for metabolism and is regulated by growth factor and nutrient signaling pathways, mediated by the Rheb and Rag GTPases, respectively. Here we provide the first animal model in which both pathways were upregulated through concurrent mutations in their GTPase-activating proteins, Tsc1 and Depdc5. Unlike former models that induced limited mTORC1 upregulation, hepatic deletion of both Tsc1 and Depdc5 (DKO) produced strong, synergistic activation of the mTORC1 pathway and provoked pronounced and widespread hepatocyte damage, leading to externally visible liver failure phenotypes, such as jaundice and systemic growth defects. The transcriptome profile of DKO was different from single knockout mutants but similar to those of diseased human livers with severe hepatitis and mouse livers challenged with oxidative stress-inducing chemicals. In addition, DKO liver cells exhibited prominent molecular pathologies associated with excessive endoplasmic reticulum (ER) stress, oxidative stress, DNA damage and inflammation. Although DKO liver pathologies were ameliorated by mTORC1 inhibition, ER stress suppression unexpectedly aggravated them, suggesting that ER stress signaling is not the major conduit of how hyperactive mTORC1 produces liver damage. Interestingly, superoxide scavengers N-acetylcysteine (NAC) and Tempol, chemicals that reduce oxidative stress, were able to recover liver phenotypes, indicating that mTORC1 hyperactivation induced liver damage mainly through oxidative stress pathways. Our study provides a new model of unregulated mTORC1 activation through concomitant upregulation of growth factor and nutrient signaling axes and shows that mTORC1 hyperactivation alone can provoke oxidative tissue injury.

Endoplasmic reticulum (ER) stress was reported to play a major role in non‐alcoholic fatty liver disease (NAFLD) induction and progression. Here, we study the effect of Zingiber officinale and omega‐3 fatty acids on ER stress for treating NAFLD. Male Wistar rats were fed on a normal diet (control group) or high‐fat diet (HFD) for 8 weeks. The HFD rats were later treated with vehicle, omega‐3 or with Z. officinale extract. HFD group demonstrated significantly more body weight gain and higher plasma lipid profile, glucose, and hepatic enzymes. The expressions of lipogenic ChREBP and ER stress genes CHOP, XBP1, and GRP78 were increased. This was accompanied by intrahepatic fat accumulation visualized by hepatic morphology and H&E‐stained sections. Treatment with Z. officinale and omega‐3 fatty acids reverted these changes into a normal healthy state. From these results, we prove that both therapeutic approaches can be potential drugs for treating NAFLD besides other ER stress‐associated diseases. Practical applications The effect of Zingiber officinale extract and omega‐3 fatty acid on ER stress associated with NAFLD was investigated. The results revealed that Z. officinale extract and omega‐3 fatty acids significantly inhibited ER stress and intrahepatic fat accumulation with the upper hand for Z. officinale extract. Both can be used as future promising therapies for the treatment of NAFLD patients and also treating different diseases that involve ER stress as a pathological modulator like diabetes mellitus, Alzheimer's disease, Parkinson's disease, and cancer.

Few publications are describing the pharmacokinetics of tylvalosin (TVN) antibiotic, so the aims of this work were to study its pharmacokinetics with evaluation the effect of vitamin (vit) E on its kinetics after single and repeated oral administration in broiler chickens. The mean serum concentrations of the drug were markedly lower in tylvalosin alone when compared with tylvalosin pre-treated with vit E after single and repeated oral administration at the corresponding time intervals. The peak of serum concentration (C max ), absorption half-life (t 0.5ab ) and elimination half-life (t 0.5 el ) were 2.11±0.3, 3.27±0.2 ug mLG 1 , 0.94±0.04, 0.88±0.02 h and 1.61±0.05, 2.42±0.1, after single tylvalosin and tylvalosin-vit E, respectively. While after repeated dosing, the maximum serum levels were 5.01±0.13 and 5.9±0.1 ug mLG 1 with pharmacokinetic parameters (C max ), (t 0.5ab ) and (t 0.5 el ) 4.1±0.14, 5.67±0.4 ug mLG 1 , 1.39±0.4, 0.86±0.07 h and 2.78±0.16, 3.05±0.3 h after repeated tylvalosin and tylvalosin-vit E, respectively. The combination of tylvalosin with vit E allows a prolongation of the dosage intervals in broiler for more 12 and 24 h after single and repeated dosing, respectively, indicated by longer elimination half-life and Mean Residence Time (MRT) of tylvalosin-vit E than tylvalosin alone after single and repeated administration which is very important for the dosage intervaland increasing the drug serum efficacy.

No abstract

Ciliopathies are a group of rare disorders characterized by a high genetic and phenotypic variability, which complicates their molecular diagnosis. Hence the need to use the latest powerful approaches to faster identify the genetic defect in these patients. We applied whole exome sequencing to six consanguineous families clinically diagnosed with ciliopathy-like disease, and for which mutations in predominant Bardet-Biedl syndrome (BBS) genes had previously been excluded. Our strategy, based on first applying several filters to ciliary variants and using many of the bioinformatics tools available, allowed us to identify causal mutations in BBS2, ALMS1 and CRB1 genes in four families, thus confirming the molecular diagnosis of ciliopathy. In the remaining two families, after first rejecting the presence of pathogenic variants in common cilia-related genes, we adopted a new filtering strategy combined with prioritisation tools to rank the final candidate genes for each case. Thus, we propose CORO2B, LMO7 and ZNF17 as novel candidate ciliary genes, but further functional studies will be needed to confirm their role. Our data show the usefulness of this strategy to diagnose patients with unclear phenotypes, and therefore the success of applying such technologies to achieve a rapid and reliable molecular diagnosis, improving genetic counselling for these patients. In addition, the described pipeline also highlights the common pitfalls associated to the large volume of data we have to face and the difficulty of assigning a functional role to these changes, hence the importance of designing the most appropriate strategy according to each case.

The Pharmacokinetic profile of thiamphenicol glycinate HCl was studied in male goats following single intravenous and intramuscular administration of 30 mg kg-1 b.wt. Thiamphenicol concentration in serum was determined by microbiological assay using Bacillus subtilis (ATCC 6633) as test organism. After intravenous injection the serum thiamphenicol concentration time course was found to obey two-compartment open model with distribution (t 0.5(α)) and elimination (t 0.5(β)) half lives of 0.0.06 ± 0.003 and 1.20 + 0.163 h., respectively. Total body clearance (Cl B) and steady state volume of distribution (Vd ss) were 1.025 ± 0.04 L kg-1 h-1 and 0.51± 0.010 L kg-1 ., respectively. After intramuscular administration the observed mean peak serum concentration (C max) was 6.89 ± 0.052 µg ml-1 achieved after maximum time (t max) of 1.53 ± 0.08 hour post-injection.The systemic bioavailability after intramuscular was 87.61 %. The plasma protein binding percent was 13.3%.

The pharmacokinetic profile of ceftiofur sodium, a third generation cephalosporin, was studied in both Friesian and buffalo calves following a single intravenous and intramuscular administration of 2.2 mg kg -1 b.wt. in a cross over study with 15-day wash out period. After i.v administration the serum concentration-time curve of ceftiofur sodium was best fitted using two-compartments open model, with distribution half-lives (t ½(α) ) of 0.384 and 0.176 h., elimination half-lives (t ½(β) ) of 5.047 and 1.607 h., mean residence time (MRT) of 6.926 and 2.072 h., volumes of distribution at steady-state (Vd ss ) of 0.206 and 0.134 L kg -1 and total body clearance (Cl B ) of 0.029 and 0.065 L kg -1 h -1 in Friesian and buffalo calves, respectively. Following intramuscular administration, the drug absorbed with half-lives of absorption (t ½(ab) ) of 1.010 and 0.217 h., maximum serum concentrations (C max ) of 5.539 and 9.663 µg ml -1 which attained after (t max ) of 3.147 and 0.825 h. and the drug was eliminated with half-lives (t ½(el) ) of 5.239 and 1.750 h. in Friesian and buffalo calves, respectively. The systemic intramuscular bioavailabilities were 89.82 and 99.7 %, while the in-vitro serum proteinbinding tendencies were 39.68 and 14.44 % in Friesian and buffalo calves, respectively.