Production performance, haematological parameters, serum biochemistry, and expression of HSP-70 in broiler chickens fed dietary ascorbic acid during heat stress.

An experiment was carried out to assess the efficacy of supplemental ascorbic acid (AA) on broiler chicken production performance, blood haematological profile, biochemical profile, and carcass traits under heat stress conditions. A total of 192-day-old broiler chicks were divided into four groups, each with six replicates of eight each (4 × 6 × 8). Four corn-based dietary treatments were formulated: T1 (control diet), T2 (T1 + AA at 200 mg/kg), T3 (T1 + AA at 400 mg/kg), and T4 (T1 + AA at 600 mg/kg) for a period of 42 days. Despite the high temperature and humidity, the 600 mg AA supplemental groups (T4) gained significantly (P ≤ 0.05) more body weight and had a higher feed intake and better feed conversion ratio (FCR) than the control group (T1). After 28 days of feeding the three AA-supplemented diets, antibody titres (humoral immune response) were significantly higher (P ≤ 0.05). The response to intradermally injected phyto-haemagglutinin (PHA-P), an index of the in vivo cell-mediated immune response, was found to be increased (P ≤ 0.05) in the 400 and 600 mg AA-supplemented groups after 35 days. Higher levels of AA (T4) supplementation significantly (P ≤ 0.05) improved haematological values such as haemoglobin (Hb), total erythrocyte count (TEC), total leukocyte count (TLC), and differential leukocyte count (DLC), heterophil:lymphocyte (H:L) in comparison to the control group (T1). The supplemented group improved the serum biochemical profile of the birds, with an increase (P ≤ 0.05) in total serum protein, albumin, and globulin and a decrease in serum cholesterol and corticosterone levels in the T4 group compared to the control group. Heat shock protein-70 (HSP-70) was gradually elevated after increasing the ascorbic acid concentration (P ≤ 0.05) at 14 and 21 days. As a result, it can be concluded that supplementing ascorbic acid at 600 mg/kg is beneficial for improving the performance, immunity, and blood haematological biochemical profile and upregulating the HSP-70 gene of broiler chickens under heat stress conditions.

Salazinic Acid and Norlobaridone from the Lichen Hypotrachyna cirrhata: Antioxidant Activity, α-Glucosidase Inhibitory and Molecular Docking Studies.

The present study was intended for the identification of secondary metabolites in acetone extract of the lichen Hypotrachyna cirrhata using UPLC-ESI-QToF-MS/MS and the detection of bioactive compounds. This study led to the identification of 22 metabolites based on their MS/MS spectra, accurate molecular masses, molecular formula from a comparison of the literature database (DNP), and fragmentation patterns. In addition, potent antioxidant and α-glucosidase inhibitory potentials of acetone extract of H. cirrhata motivated us to isolate 10 metabolites, which were characterized as salazinic acid (11), norlobaridone (12), atranorin (13), lecanoric acid (14), lichesterinic acid (15), protolichesterinic acid (16), methyl hematommate (17), iso-rhizonic acid (18), atranol (19), and methylatratate (20) based on their spectral data. All these isolates were assessed for their free radicals scavenging, radical-induced DNA damage, and intestinal α-glucosidase inhibitory activities. The results indicated that norlobaridone (12), lecanoric acid (14), methyl hematommate (17), and atranol (19) showed potent antioxidant activity, while depsidones (salazinic acid (11), norlobaridone (12)) and a monophenolic compound (iso-rhizonic acid, (18)) displayed significant intestinal α-glucosidase inhibitory activities (p < 0.001), which is comparable to standard acarbose. These results were further correlated with molecular docking studies, which indicated that the alkyl chain of norlobaridione (12) is hooked into the finger-like cavity of the allosteric pocket; moreover, it also established Van der Waals interactions with hydrophobic residues of the allosteric pocket. Thus, the potency of norlobaridone to inhibit α-glucosidase enzyme might be associated with its allosteric binding. Also, MM-GBSA (Molecular Mechanics-Generalized Born Surface Area) binding free energies of salazinic acid (11) and norlobaridone (12) were superior to acarbose and may have contributed to their high activity compared to acarbose.

Changes in m6A RNA methylation of goat lung following PPRV infection.

Peste des petits ruminants (PPR) is an acute, highly contagious viral disease of goats and sheep, caused by the Peste des petits ruminants virus (PPRV). Earlier studies suggest the involvement of diverse regulatory mechanisms in PPRV infection. Methylation at N6 of Adenosine called m6A is a type RNA modification that influences various physiological and pathological phenomena. As the lung tissue represents the primary target organ of PPRV, the present study explored the m6A changes and their functional significance in PPRV disease pathogenesis. m6A-seq analysis revealed 1289 m6A peaks to be significantly altered in PPRV infected lung in comparison to normal lung, out of which 975 m6A peaks were hypomethylated and 314 peaks were hypermethylated. Importantly, hypomethylated genes were enriched in Interleukin-4 and Interleukin-13 signaling and various processes associated with extracellular matrix organization. Further, of the 843 differentially m6A-containing cellular transcripts, 282 transcripts were also found to be differentially expressed. Functional analysis revealed that these 282 transcripts are significantly enriched in signaling by Interleukins, extracellular matrix organization, cytokine signaling in the immune system, signaling by receptor tyrosine kinases, and Toll-like Receptor Cascades. We also found m6A reader HNRNPC and the core component of methyltransferase complex METTL14 to be highly upregulated than the m6A readers - HNRNPA2B1 and YTHDF1 at the transcriptome level. These findings suggest that alteration in the m6A landscape following PPRV is implicated in diverse processes including Interleukin signaling.

First report of whole genome sequence of septicemic Pasteurella multocida serovar B:2 'Soron' strain isolated from swine.

Pig pasteurellosis, caused by Pasteurella multocida, is an acute infection that also has economic implications for pig farmers. We report the complete genome sequence of a P. multocida, serovar B:2 'Soron' strain isolated from the blood of a pig that had died of pasteurellosis in India. The isolate was not found to be haemorrhagic septicaemia (HS) specific B:2 by the PCR assay. The genome of 'Soron' strain is a single circular chromosome of 2,272,124 base pairs in length and contains 2014 predicted coding regions, 4 ribosomal RNA operons, and 52 tRNAs. It has 1812 protein-coding genes that were also found in reference sequence PmP52Vac. Phylogenetic analysis revealed that Pm_P52VAc and P. multocida 'Soron' serovar B:2 were clustered in different clades. Pasteurella multocida 'Soron' serovar B:2 was found to cluster with the same ancestor of Pm70, which is of avian origin. The genome was found to contain regions that encode proteins which may confer resistance to various antibiotics including cephalosporin, which is used to treat pasteurellosis. The isolate was also found to harbour a phage region. This strain represents a novel multi-locus sequence type (MLST) that has not been previously identified, as all of the alleles used for MLST were found, but did not match any of the alleles in the database with 100% nucleotide identity. The most closely related ST was ST221. This is the first whole-genome sequence from P. multocida serovar B:2 of pig origin.

Effects of Various Copper Sources and Concentrations on Performance, Skeletal Growth, and Mineral Content of Excreta in Broiler Chickens.

The experiment was designed to study the effect of supplemental sources and concentrations of copper on the performance and development and mineralization of tibia bones in broiler chickens. A 42-day feeding experiment was conducted utilising three copper sources, including copper sulphate (CuS), copper chloride (CuCl), and copper propionate (CuP), each with four different concentrations, i.e. 8, 100, 150, and 200 mg/kg. The body weight gain with 200 mg Cu/kg food was noticeably higher during the first 4-6 weeks of age. Due to the interaction between Cu sources and levels, there was no significant change in the body weight gained. The feed intake during various growing phases did differ significantly neither the main effect nor the interaction between different copper sources and levels. A CuP-supplemented diet (200 mg/kg food) considerably (P ≤ 0.05) improved the feed conversion ratio between 4-6 and 0-6 weeks. At the end of the experiment, a total of 72 tibia bones, i.e. six for each treatment were collected. A metabolic trial was conducted to look into mineral retention in broiler chickens on the final 3 days of the trial (40-42 days). Increased tibia bone zinc (Zn) levels were seen with the addition of 8 mg Cu/kg of Cu chloride, 100 mg Cu/kg of Cu propionate, 8 mg Cu/kg of Cu sulphate, and 8 mg/kg of Cu propionate to the diet. At higher levels of Cu (150 and 200 mg/kg diet), there was a significantly (P ≤ 0.01) reduced tibia Zn content. Cu sulphate treatment group had higher (P ≤ 0.01) tibia Cu content (8 mg Cu/kg diet). Cu sulphate supplemented diet had a greater excreta Zn content (P ≤ 0.01) than Cu chloride supplemented diet, and Cu propionate supplemented diet had the lowest excreta Zn content. Excreta with a higher Fe concentration were found in diets supplemented with copper sulphate and copper chloride (P ≤ 0.05) than in diets supplied with copper propionate. Thus, it may be concluded that feeding dietary Cu concentrations up to 200 mg Cu/kg diet, regardless of the different sources, had no negative effects on bone morphometry and mineralization parameters with the exception of a decrease in the tibia's zinc content.

Effects of Different Concentration of Copper on Performance, Immunity and Carcass Traits in Broiler Japanese Quails.

In this study, the effects of dietary copper supplements on broiler Japanese quail growth performance, immune response, blood biochemistry, and carcass quality were examined. Two copper sources (copper sulphate-CuS, and copper methionine-CuM), each at five distinct dietary dosages of 5, 10, 15, 100, and 150 mg/kg, were used. A total of 280 (10 × 4 × 7) day-old quail chicks of uniform body weight were randomly distributed into 10 treatments with 4 replicates each and having 7 chicks in each replicate. In comparison to CuS-supplemented diets, CuM-supplemented diets (100 mg Cu/kg diet) considerably (P ≤ 0.01) increased body weight gain and improved feed conversion ratio (FCR). In the 150-mg CuM/kg diet, the cell-mediated immune response (foot web index to PHAP) was considerably (P ≤ 0.01) greater. The humoral immune response (HA titre to SRBC) was substantially (P ≤ 0.01) lower with CuS-supplemented meals than with CuM-supplemented diets. When compared to CuS source, the weight of the bursa and spleen from CuM source was considerably (P ≤ 0.01) higher. The 100- and 150-mg CuM/kg diets considerably (P ≤ 0.01) reduced serum cholesterol levels. Thus, it may be concluded that dietary supplementation of copper methionine as a source of Cu @ 100 mg Cu/kg diet to broiler Japanese quails was more effective in improving growth performance, immunological response, carcass quality features, and serum cholesterol reduction.

Effects of paraprobiotic as replacements for antibiotic on performance, immunity, gut health and carcass characteristics in broiler chickens.

This study sought to determine the effects of dietary paraprobiotic (PPB) on broiler chicken performance, immunity, gut health, and carcass traits. A total of 240 day-old CARIBRO Vishal commercial broiler chicks of identical body weight randomly divided into six treatment groups, each with five replicates and eight chicks in each replicate. Six dietary treatments were preapared: T1 = (control diet), T2 = T1 + 0.02% (w/v) chlortetracycline (CTC), T3 = T1 + 0.2% (w/v) PPB, T4 = T1 + 0.4% (w/v) PPB, T5 = T1 + 0.6% (w/v) PPB and T6 = T1 + 0.8% (w/v) PPB, respectively. Body weight gain (BWG) significantly (P ≤ 0.05) increased in the T5 (0.6% PPB) and T6 (0.8% PPB) group. At the same time the feed intake significantly (P ≤ 0.05) decreased and the feed conversion ratio (FCR) significantly (P ≤ 0.05) improved in T5 and T6 group. There was a significant (P ≤ 0.05) increase in cell-mediated immunity and haem-agglutination titre (HA titre) in the 0.6% and 0.8% PPB supplemented groups compare to the control group (T1). The percentage of carcass traits and organ weights did not significantly differ between the PPB-supplemented and control groups, but the percentage of live weight in cut up parts showed a significant improvement (P ≤ 0.05) in the PPB-supplemented group. At 42 days, villus height, width, and crypt depth all significantly (P ≤ 0.05) increased in the groups supplemented with 0.6 and 0.8% para-probiotics (T5 and T6). The results show that para-probiotics can be added to broiler diets at a rate of 0.6% (w/v) to enhance performance, immunity, gut health, and breast yield. The para-probiotic may therefore be a useful substitution for antibiotic growth promoters in the diet of chickens.

Comprehensive Lichenometabolomic Exploration of Ramalina conduplicans Vain Using UPLC-Q-ToF-MS/MS: An Identification of Free Radical Scavenging and Anti-Hyperglycemic Constituents.

In this study, we propose ultra-performance liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UPLC-QToF-MS/MS)-guided metabolite isolation as a choice analytical approach to the ongoing structure−activity investigations of chemical isolates from the edible lichen, Ramalina conduplicans Vain. This strategy led to the isolation and identification of a new depside (5) along with 13 known compounds (1−4, 6−14), most of which being newly described in this lichen species. The structures of the isolates were established by detailed analysis of their spectral data (IR, NMR, and Mass). The acetone extract was further analyzed by UPLC-Q-ToF-MS/MS in a negative ionization mode, which facilitated the identification and confirmation of 18 compounds based on their fragmentation patterns. The antioxidant capacities of the lichen acetone extract (AE) and isolates were measured by tracking DPPH and ABTS free radical scavenging activities. Most isolates displayed marked radical scavenging activities against ABTS while moderate activities were observed against DPPH radical scavenging. Except for atranol (14), oxidative DNA damage was limited by all the tested compounds, with a marked protection for the novel isolated compound (5), as previously noted for the acetone extract (p < 0.001). Furthermore, compound (4) and acetone extract (AE) have inhibited intestinal α-glucosidase enzyme significantly (p < 0.01). Although some phytochemical studies were already performed on this lichen, this study provided new insights into the isolation and identification of bioactive compounds, illustrating interest in future novel analytical techniques.

Chemical profiling of marine seaweed Halimeda gracilis using UPLC-ESI-Q-TOF-MSE and evaluation of anticancer activity targeting PI3K/AKT and intrinsic apoptosis signaling pathway.

Marine seaweeds are predominant for nutraceuticals and have been food source since ancient times and are currently being used in Chinese medicines and Japanese traditional medicines. In the present study, the chemical profile analysis of Halimeda gracilis was performed using UPLC-ESI-Q-TOF-MSE analysis and assessed for its anticancer activity. A cursory investigation of the total ion chromatograms of the both methanol (MHG) and ethyl acetate (EAHG) of H. gracilis extracts reveals that both extracts have different kind of metabolites including phenols and its derivatives, diterpenes, cinnamic acids derivatives etc. The in vitro anticancer activity of EAHG and MHG exhibiting significant activity and induced apoptosis against skin cancer cells by generating excessive ROS, damaging the mitochondrial membrane and nuclear components. Further, the western blot analysis showed that the EAHG and MHG downregulates the oncoproteins PI3K, AKT, p-AKT, and BcL2 and upregulates the apoptotic proteins Bax, Cyto C, p21, p53, Caspase 9 and Caspase 3. To best of our knowledge, this is the first report which implies the UPLC-ESI-Q-TOF-MSE based chemical profiling of H. gracilis and can be used for the treatment of cancer.

Bombax ceiba calyx displays antihyperglycemic activity via improving insulin secretion and sensitivity: Identification of bioactive phytometabolomes by UPLC-QTof-MS/MS.

Vegetables are considered good food for the management of hyperglycemia. Bombax ceiba L. (family: Bombacaceae) calyces are part of traditional vegetables. This study evaluated its usefulness on various parameters responsible for the development of hyperglycemia and conducted phytometabolomic analysis to identify phytochemicals responsible for the observed activities. It was found that the aqueous methanol extract of its calyces (B. ceiba calyx extract, BCE) reduced (12.4%) significantly (p < 0.05) the development of sucrose-induced postprandial hyperglycemic load in rats. In-vitro studies revealed that BCE improved glucose-stimulated insulin secretory activity in MIN6 cells plausibly by decreasing ADP/ATP ratio. BCE also augmented concentration-dependent (5 µg, 10 µg, and 20 µg) increase in glucose uptake in hyperglycemic L6 myotubes both by non-insulin-dependent manner (35%, 68%, and 132%, respectively) and insulin-dependent manner (42%, 59%, and 172%, respectively). The insulin-stimulated GLUT4 translocation was compromised (34%) significantly (p < 0.05) under hyperglycemic condition; however, it was improved by 23% and 72% (p < 0.001) when L6 myotubes were primed with 10 and 20 µg of BCE, respectively. Hyperglycemia aggravated reactive oxygen species (ROS) generation in L6 myotubes. The ROS generation was significantly (p < 0.001) reduced by priming myotubes with BCE before challenging myotubes to hyperglycemic environment, possibly by preserving cellular antioxidant enzymes catalase, glutathione peroxidase, and reduced glutathione levels. Phytometabolomic analysis disclosed a number of phytochemicals present in B. ceiba calyces known to display these activities. This is the first study reporting antihyperglycemic activity in B. ceiba calyces, its mechanisms of action, and phytometabolomic profile applying UPLC-QTof-MS/MS technique. PRACTICAL APPLICATION: B. ceiba calyces are part of traditional vegetables. Our study finds that B. ceiba calyces contain phytochemicals possessing antihyperglycemic, insulin secretory, insulin sensitization properties, and potentials for preserving hyperglycemia-induced vitiations in cellular antioxidant defense. These observations provide foundation for exploring further possibilities of B. ceiba calyces to become valuable dietary inclusion in the diet of people suffering from metabolic disorders.

Effect of dietary mannan oligosaccharides and fructo-oligosaccharides on physico-chemical indices, antioxidant and oxidative stability of broiler chicken meat.

The objective of this present study was to investigate the potentiality of prebiotics (mannan oligosaccharides-MOS and fructo-oligosaccharides-FOS) in replacement of antibiotic growth promoter and their relationship with physico-chemical indices, antioxidant and oxidative stability and carcass traits of broiler chickens meat. Accordingly, 240 day-old broiler chicks of uniform body weight divided in 6 treatment groups with 5 replicate each (5 × 6 = 30) having 8 birds in each replicate. Six corn based dietary treatments were formulated viz. T1 (control diet), T2 (T1 + Bacitracin methylene di-salicylate @ 0.002%), T3 (T1 + 0.1% MOS), T4 (T1 + 0.2% MOS), T5 (T1 + 0.1% FOS), and T6 (T1 + 0.2% FOS). Significant (p < 0.05) increase in cut up part yields (%) and reduction in cholesterol and fat content in T4 (0.2% MOS) group. The water holding capacity (WHC) and extract release volume (ERV) were increase (p < 0.05) in 0.1 or 0.2% MOS supplemented group. DPPH (1, 1-diphenyl-2-picrylhydrazy) was higher (p < 0.05) and lipid oxidation (free fatty acid and thio-barbituric acid reactive substances) was lower (p < 0.05) in T4 group. The standard plate count (SPC), staphylococcus and coliform counts were decreased (p < 0.05) in T3 or T4 group. Thus, it can be concluded that mannan oligosaccharides (MOS) may be incorporated at 0.2% level in diet for improved physico-chemical indices, antioxidant and oxidative stability and carcass characteristics of broiler chickens meat and it may be suitable replacer of antibiotic growth promoter.

Genetic diversity in the Tams1 gene of Theileria annulata (Duschunkowsky and Luhs, 1904) infecting cattle.

The present study describes the genetic diversity in the Tams1 gene (733 bp) of Theileria annulata along with the sequence, phylogenetic and haplotype analyses of the Indian isolates. The phylogenetic analyses displayed distinct clustering of the Indian isolates into three groups suggesting the presence of three genotypes, hitherto designated as T. annulata genotypes 1-3 (G1-G3). Genotype 3 seems to be novel containing only two newly generated sequences. Indian isolates displayed 88.4-100% and 82.2-100% similarity with each other at nucleotide (nt) and amino acid (aa) levels, respectively. However, the newly generated sequences (n = 36) showed 90.5-100% and 84.3-100% identity between them at nt and aa levels, respectively. The most diverse and heterogeneous genotype, G1, exhibited the highest number of polymorphic sites (S = 148), haplotypes (h = 16) and nucleotide differences (k = 43.23) besides haplotype (Hd = 0.903 ± 0.031) and nucleotide (π = 0.059 ± 0.005) diversities. Neutrality indices suggested a respective decrease and increase in population sizes of G1 and G2 genotypes in India. The nucleotide sequence analyses indicated the presence of extensive sequence variations between nucleotide positions 1-124, 194-257 and 396-494. The N-terminus of Tams1 protein displayed a considerable sequence variability with extensive variations in two regions, between amino acid positions 1-39 and 127-172, as compared to the conserved carboxyl terminus.

Systems biology under heat stress in Indian cattle.

Transcriptome profiling of Vrindavani and Tharparkar cattle (n = 5 each) revealed that more numbers of genes were dysregulated in Vrindavani than in Tharparkar. A contrast in gene expression was observed with 18.9 % of upregulated genes in Vrindavani downregulated in Tharparkar and 17.8% upregulated genes in Tharparkar downregulated in Vrindavani. Functional annotation of genes differentially expressed in Tharparkar and Vrindavani revealed that the systems biology in Tharparkar is moving towards counteracting the effects due to heat stress. Unlike Vrindavani, Tharparkar is not only endowed with higher expression of the scavengers (UBE2G1, UBE2S, and UBE2H) of misfolded proteins but also with protectors (VCP, Serp1, and CALR) of naïve unfolded proteins. Further, higher expression of the antioxidants in Tharparkar enables it to cope up with higher levels of free radicals generated as a result of heat stress. In this study, we found relevant genes dysregulated in Tharparkar in the direction that can counter heat stress.

Canine Parvovirus and Its Non-Structural Gene 1 as Oncolytic Agents: Mechanism of Action and Induction of Anti-Tumor Immune Response.

The exploration into the strategies for the prevention and treatment of cancer is far from complete. Apart from humans, cancer has gained considerable importance in animals because of increased awareness towards animal health and welfare. Current cancer treatment regimens are less specific towards tumor cells and end up harming normal healthy cells. Thus, a highly specific therapeutic strategy with minimal side effects is the need of the hour. Oncolytic viral gene therapy is one such specific approach to target cancer cells without affecting the normal cells of the body. Canine parvovirus (CPV) is an oncolytic virus that specifically targets and kills cancer cells by causing DNA damage, caspase activation, and mitochondrial damage. Non-structural gene 1 (NS1) of CPV, involved in viral DNA replication is a key mediator of cytotoxicity of CPV and can selectively cause tumor cell lysis. In this review, we discuss the oncolytic properties of Canine Parvovirus (CPV or CPV2), the structure of the NS1 protein, the mechanism of oncolytic action as well as role in inducing an antitumor immune response in different tumor models.

Phytometabolomic analysis of boiled rhizome of Nymphaea nouchali (Burm. f.) using UPLC-Q-TOF-MSE, LC-QqQ-MS & GC-MS and evaluation of antihyperglycemic and antioxidant activities.

Phytometabolomic analysis of Nymphaea nouchali (Burm. F.) boiled rhizome was carried out utilizing UPLC-Q-TOF-MSE, LC-QqQ-MS and GC-MS techniques and evaluated for antihyperglycemic and antioxidative stress potentials. Metabolomic analysis revealed presence of multiple antidiabetic and antioxidant compounds. Boiled rhizome powder exhibited potent antihyperglycemic activity against sugar-induced postprandial hyperglycemia in rats plausibly due to the presence of intestinal α-glucosidase inhibitory and augmenting cellular glucose uptake activities. It also prevented hyperglycemia-induced hemoglobin and insulin glycation. Rhizome displayed potent reducing power, effectively scavenged various reactive oxygen species. It displayed antioxidative stress potential in assuaging H2O2 induced erythrocyte hemolysis and antioxidant activity by inhibiting membrane lipid peroxidation. Boiled rhizome was also found to preserve the loss of cellular antioxidants under H2O2 induced oxidative stress and disturbances caused to mitochondrial membrane potential. This is the first research reporting boiled N. nouchali rhizome as an ideal food material to manage the cause of hyperglycemia and resultant oxidative stress.

Evaluation of Diagnostic Accuracy of Magnetic Resonance Imaging in Posterior Ligamentum Complex Injury of Thoracolumbar Spine.

STUDY DESIGN: Prospective diagnostic imaging study. PURPOSE: The stability of the thoracic and lumbar spine depends significantly on the posterior ligamentum complex (PLC). Therefore, it is essential to diagnose PLC injuries accurately before deciding on a treatment plan for thoracolumbar injury patients. However, the efficacy of magnetic resonance imaging (MRI) in diagnosing PLC remains undetermined. OVERVIEW OF LITERATURE: MRI has become the ultimate tool in diagnosing spine injury cases, as previous literature suggests that it has very high sensitivity and specificity. But this is still controversial and as many surgeons rely on just MRI for selecting the patient for surgery, it becomes important to know the diagnostic accuracy of it. METHODS: Patients who sustained injuries from T1 to L3 and required posterior surgery were prospectively studied. The treating surgeon and musculoskeletal radiologist participating in the study reviewed preoperative MRI images to characterize the level(s) of injury and the integrity of the six components of the PLC. These were classified as intact, incompletely disrupted, or disrupted. During the surgical procedure, the surgeon also classified each component of the PLC, and the radiologist's and surgeon's findings were compared. RESULTS: Out of 66 patients, 46 were males (69.7%) and 20 were females (30.3%), and the average age was 34.12 years. According to the kappa score, there was a moderate level of agreement between the radiologist's interpretation and the intraoperative findings for all PLC components except for the thoracolumbar fascia and ligamentum flavum for which there was a slight agreement. The sensitivity for the intact PLC components ranged from 100% (supraspinous ligament) to 66.67% (ligamentum flavum). The specificity ranged from 100% (interspinous ligament) to 52% (thoracolumbar fascia). The Spearman's rank correlation ranged from 0.061 for the thoracolumbar fascia to 0.918 for the interspinous ligament, and the percentage agreement ranged from 81.82% (interspinous ligament to 36.36% (thoracolumbar fascia). CONCLUSIONS: The sensitivity and specificity of MRI for diagnosing injury of the PLC in this study were lower than those previously reported in the literature. The integrity of the PLC as determined by MRI should not be used in isolation to determine treatment.

A novel quantitative assay for analysis of GLUT4 translocation using high content screening.

Insulin resistance is associated with obesity and can lead to several metabolic disorders including type II diabetes, nonalcoholic fatty liver disease and cardiovascular problems. Search for the small molecules which can either induce or mimic the insulin action are of great interest and can be utilized to manage insulin resistance. There are several dietary phytochemicals which can potentially have insulinomimetic action. Nevertheless, high throughput screening methods to test efficiency of small molecules to act as an insulinomimetic are not fully established. In this paper we have performed chemical screen analysis based on GLUT4 translocation using a cell line CHO-HIRC-myc-GLUT4 eGFP that expresses GLUT4-GFP in association with human Insulin receptor. We have established a high content screening-based method which can track and quantify the GLUT4 translocation from perinuclear area to the cell membrane. The assay involves measuring fluorescence intensity in a defined perinuclear area and a defined area along the cell membrane; and the results are expressed as the ratio of fluorescence intensity in the perinuclear to membrane area. The assay could collect real time data of GLUT4 translocation from thousand of cells/ sample and from many such samples in one experiment. We validated the assay using Insulin, insulin mimics/sensitizers and insulin inhibitors. The agonist or antagonists were analyzed for their ability to enhance or block the GLUT4 translocation independent of insulin. The outcome of the assay was correlated by performing glucose uptake assay using differentiated 3T3L1 cells. Using this platform we further identified several plant extracts which had the insulin mimetic action. We confirmed that these plant extracts were non-toxic to the beta cells using RIN mf5cells and 3T3L1 cells. We have identified plant extracts with the potential insulinomimetic action using novel high-content screening approach; these can be further tested for their efficiency in-vivo in pre-clinical trials.

Stability of live attenuated classical swine fever cell culture vaccine virus in liquid form for developing an oral vaccine.

Classical swine fever (CSF) is an important viral disease of pigs and controlled by vaccination. Unorganised backyard and wild pigs are difficult to vaccinate by needle vaccination. Here we formulated liquid vaccines using an Indian CSF cell culture vaccine virus and four stabilisers and evaluated their stability at 4 °C, 25 °C and 37 °C up to 24 h for use as oral vaccine. The stabilisers were Lactalbumin hydrolysate-Trehalose, Lactalbumin hydrolysate-Trehalose-Gelatin, Lactalbumin hydrolysate-Lactose-Sucrose and Lactalbumin hydrolysate-Sucrose. The liquid vaccines, with or without stabilisers, were stable at 4 °C up to 24 h, whereas, a drop of one log10 titre was observed at 25 °C during the same period. At 37 °C, the virus titre diminished by only one log10 with the Lactalbumin hydrolysate-Trehalose (LT) stabiliser up to 24 h compared to two log10 losses in virus titre with other stabilisers and virus control. We therefore conclude that for developing a CSF oral vaccine, the vaccine virus in liquid form can be used directly during the winter, whereas for developing the oral vaccine for summer, the LT stabiliser would provide maximum stability to the virus to withstand the warm temperature while maintaining adequate therapeutic titre for inducing a protective immune response.

Apoptin as a Tumor-Specific Therapeutic Agent: Current Perspective on Mechanism of Action and Delivery Systems.

Cancer remains one of the leading causes of death worldwide in humans and animals. Conventional treatment regimens often fail to produce the desired outcome due to disturbances in cell physiology that arise during the process of transformation. Additionally, development of treatment regimens with no or minimum side-effects is one of the thrust areas of modern cancer research. Oncolytic viral gene therapy employs certain viral genes which on ectopic expression find and selectively destroy malignant cells, thereby achieving tumor cell death without harming the normal cells in the neighborhood. Apoptin, encoded by Chicken Infectious Anemia Virus' VP3 gene, is a proline-rich protein capable of inducing apoptosis in cancer cells in a selective manner. In normal cells, the filamentous Apoptin becomes aggregated toward the cell margins, but is eventually degraded by proteasomes without harming the cells. In malignant cells, after activation by phosphorylation by a cancer cell-specific kinase whose identity is disputed, Apoptin accumulates in the nucleus, undergoes aggregation to form multimers, and prevents the dividing cancer cells from repairing their DNA lesions, thereby forcing them to undergo apoptosis. In this review, we discuss the present knowledge about the structure of Apoptin protein, elaborate on its mechanism of action, and summarize various strategies that have been used to deliver it as an anticancer drug in various cancer models.

Bombax ceiba (Linn.) calyxes ameliorate methylglyoxal-induced oxidative stress via modulation of RAGE expression: identification of active phytometabolites by GC-MS analysis.

Non-enzymatic reactions between proteins and methylglyoxal (MG) result in the formation of advanced glycation end products (AGEs). These AGEs play a vital role in the development of diabetic complications by stimulating oxidative stress and acting upon their receptor RAGE (Receptor for Advanced Glycation End products). This study examined the effect of aqueous methanol extract of Bombax ceiba L. calyxes (BCCE) on MG induced protein glycation and oxidative stress, followed by the identification of phytometabolites present in the calyxes using gas chromatography-mass spectrometry (GC-MS). The study revealed that priming of bovine serum albumin protein with the BCCE inhibited MG induced AGE formation in vitro and restrained AGE-induced RAGE up-regulation in HEK-293 cells. The BCCE significantly (p < 0.001) reduced the MG induced increase in reactive oxygen species (ROS), NADPH oxidase (NOX), and mitochondrial dysfunction. Improvements in the levels of antioxidant enzymes such as Mn and Cu/Zn-superoxide dismutase and glutathione reductase were also observed in HEK-293 cells. Furthermore, the decrease in primary cellular defense against AGEs, the glyoxalase 1 (Glo-1) activity, due to MG treatment was restored in BCCE treated cells. GC-MS analysis revealed the presence of antioxidant and antiglycation compounds such as myo-ionisitol, scopoletin, d-sedoheptulose, succinic acid, and xylitol in B. ceiba calyxes. The observed beneficial effect in our study might be attributed to the presence of these compounds in B. Ceiba calyxes. This is the first report presenting the antioxidant and antiglycation activities of B. ceiba calyxes and GC-MS analysis of active phytometabolites. These observations show that B. ceiba calyxes may become a potent and promising functional food to manage/control the development of diabetic complications.