Metabolomics-a powerful tool in livestock research.

Advancements in the Nuclear Magnetic Resonance (NMR) and Mass Spectrometry (MS) along with recent developments in omics sciences have resulted in a better understanding of molecular mechanisms and pathways associated with the physio-pathological state of the animal. Metabolomics is a post-genomics tool that deals with small molecular metabolites in a given set of time which provides clear information about the status of an organism. Recently many researchers mainly focus their research on metabolomics studies due to its valuable information in the various fields of livestock management and precision dairying. The main aim of the present review is to provide an insight into the current research output from different sources and application of metabolomics in various areas of livestock including nutri-metabolomics, disease diagnosis advancements, reproductive disorders, pharmaco-metabolomics, genomics studies, and dairy production studies. The present review would be helpful in understanding the metabolomics methodologies and use of livestock metabolomics in various areas in a brief way.

Biological methylation: redefining the link between genotype and phenotype.

The central dogma of molecular biology is responsible for the crucial flow of genetic information from DNA to protein through the transcription and translation process. Although the sequence of DNA is constant in all organs, the difference in protein and variation in the phenotype is mainly due to the quality and quantity of tissue-specific gene expression and methylation pattern. The term methylation has been defined and redefined by various scientists in the last fifty years. There is always huge excitement around this field because the inheritance of something is beyond its DNA sequence. Advanced gene methylation studies have redefined molecular genetics and these tools are considered de novo in alleviating challenges of animal disease and production. Recent emerging evidence has shown that the impact of DNA, RNA, and protein methylation is crucial for embryonic development, cell proliferation, cell differentiation, and phenotype production. Currently, many researchers are focusing their work on methylation to understand its significant role in expression, disease-resistant traits, productivity, and longevity. The main aim of the present review is to provide an overview of DNA, RNA, and protein methylation, current research output from different sources, methodologies, factors responsible for methylation of genes, and future prospects in animal genetics.

Defaecation disorders among information technology personnel: A cross-sectional study.

Background Individuals working in the information technology (IT) industry are likely to develop lifestyle disorders. We aimed to determine the presence of defaecation-related disorders in IT personnel in Chennai. Methods This cross-sectional, questionnaire-based study was done from June to December 2018. We included employees between 18 and 60 years of age and excluded those with <1 year stay in Chennai, pregnancy and those who sent incomplete responses. The cohort was classified as normal, irritable bowel syndrome (IBS), faecal evacuation disorder (FED) and a combination of the latter two. Results The overall response rate was 95.6%. The study included 54.7% of men, and the median age was 31 years. The majority of respondents used an Indian toilet (554; 58%). Almost all (96.2%) passed stools daily, and stool consistency was soft in 59%. Based on the study criteria, 180 (18.8%) had FED, 56 (5.9%) had IBS and a small group had a combination of symptoms of IBS and FED (20; 2.1%). Respondents with symptoms of IBS had a higher proportion of comorbid states (p<0.0001), lesser stool frequency (p<0.0001) and required more toilet time (p<0.00001). Conclusion Over one-fourth (26.7%) of the respondents had defaecation-related issues, 18.8% had symptoms suggestive of FED and 5.6% that of IBS, often above 30 years of age.

Healthy broilers disseminate antibiotic resistance in response to tetracycline input in feed concentrates.

Wide varieties of antibiotics are used in poultry farms to improve the growth and also to control the infection in broiler chicken. To identify the seriousness of the same in the poultry sector, current study has been designed to analyze the presence of tetracycline in poultry feed and also the tetracycline resistance among the bacteria released through the excreta of poultry. In the study, 27 bacteria belonging to the Escherichiacoli and Klebsiellapneumoniae. were isolated from the faecal samples collected from five different farms. Antibiotic susceptibility analysis showed 77% of E. coli and 100% of the K. pneumoniae. to be resistant to tetracycline. Further, molecular screening for tetA and tetB genes showed 85.18% of isolates to have tetA and 22.22% with tetB. The presence of tetracycline in collected feed samples was also analysed quantitatively by Liquid chromatography-mass spectrometry (LC-MS). Here, three out of five feed samples were found to be positive for tetracycline. The study showed a direct correlation between the antibiotic supplemented feed and the emergence of antimicrobial resistance among the intestinal microflora. The results of the study indicate the need for strict control over antibiotic use in animal feed to limit the rapid evolution and spread of antimicrobial resistance.

Label-free quantitative proteomics analysis reveals distinct molecular characteristics in endocardial endothelium.

Endocardial endothelium, which lines the chambers of the heart, is distinct in its origin, structure, and function. Characterization studies using genomics and proteomics have reported molecular signatures supporting the structural and functional heterogeneity of various endothelial cells. However, though functionally very important, no studies at protein level have been conducted so far characterizing endocardial endothelium. In this study, we used endothelial cells from pig heart to investigate if endocardial endothelial cells are distinct at the proteome level. Using a high-throughput liquid chromatography-tandem mass spectrometry for proteome profiling and expression, we identified sets of proteins that belong to specific biological processes and metabolic pathways in endocardial endothelial cells supporting its specific structural and functional roles. The study also identified several transcription factors and cell surface markers, which may have roles in the specificity of endocardial endothelium. The detection of sets proteins preferentially expressed in endocardial endothelium offers new insights into its role in the regulation of cardiac function. Data are made available through ProteomeXchange with identifier PXD009194.

Immunohistochemical Expression of Cathepsin D in Primary and Recurrent Squamous Cell Carcinoma.

AIM: The aim of this study is to analyze and compare the immunohistochemical expression of cathepsin B in primary oral squamous cell carcinoma (OSCC) and recurrent OSCC. MATERIALS AND METHODS: A total of 50 cases were studied immunohistochemically for rabbit polyclonal antihuman cathepsin D expression. A total of 10 cases of breast carcinoma were taken as positive controls. Immunohistochemical staining was performed using labeled streptavidin-biotin technique. RESULTS: All the 45 cases of OSCC, both primary and recurrent cases included, showed varying grades of cathepsin D immu-noreactivity. Statistical significance at 5% level was observed in cathepsin D expression between the different grades of well, moderate, and poorly differentiated primary squamous cell carcinomas. In the comparison of cathepsin D staining intensity among primary squamous cell carcinomas with and without recurrence, a statistical significance between the groups was observed when the p-value was at 10%, but the same comparison was not significant when the p-value was at 5%. CONCLUSION: Cathepsin D expression in primary squamous cell carcinomas with recurrences was very variable as compared with primary squamous cell carcinomas without recurrences. Comparison of cathepsin D expression in primary with their recurrent counterparts showed mostly similar intensity of expression in recurrent carcinomas, thus suggesting its limited usefulness in predicting recurrence. CLINICAL SIGNIFICANCE: Although cathepsin D might have shown limited usefulness in predicting cancer recurrence, it, however, is a proven valuable tool to detect the aggressiveness of various other tumors, and if corroborated with a larger sample may hold the key to early, more effective, and more specific treatment modalities for cases of oral cancer also.

Proteomic Signature of Endothelial Dysfunction Identified in the Serum of Acute Ischemic Stroke Patients by the iTRAQ-Based LC-MS Approach.

Acute ischemic stroke (AIS) is a devastating cerebrovascular disorder that leads to permanent physical and neurological disabilities in adults worldwide. Proteins associated with stroke pathogenesis may appear in the serum of AIS patients due to blood-brain barrier dysfunction, thus permitting the development of blood-based biomarkers for early diagnosis of stroke. These biomarkers could perhaps be an adjunct to the existing imaging modalities and aid in better management and therapeutic intervention during the course of the disease. For this exploratory study, a combination of multiplexed isobaric tagging using iTRAQ reagents and high resolution tandem mass spectrometry was used to identify differentially expressed proteins in serum samples from AIS patients. The quantitative proteomic analysis of serum from both AIS and control subjects revealed 389 high confidence protein identifications and their relative levels. Among them, 60 proteins showed a ≥1.5-fold change in the AIS subjects. We verified the altered serum levels of candidate proteins such as vWF, ADAMTS13, S100A7, and DLG4 through ELISA, and the results also corroborate with the experimental findings. vWF and ADAMTS13 are key players that regulate blood hemostasis, and their altered concentration may contribute to endothelial dysfunction. S100A7 is a novel candidate protein identified in this study that is also known to mediate inflammation, endothelial proliferation, and angiogenesis. The current study provided a potential and novel biomarker panel that may in turn provide diagnostic aid to the existing imaging modalities for the rapid diagnosis of ischemic stroke.

The rupture of a single liquid aluminium alloy film.

The present study is based on the idea of understanding the rupture of films in metal foams by studying free standing metallic films as a model system. Liquid dynamics, the velocity of the rupturing material as well as the behaviour of ceramic particles inside the melt were analysed optically ex situ and by synchrotron X-ray radiography in situ. It was found that the resistance of films to rupture is mainly based on the interaction between solid particles and an immobile oxide skin, the formation of which depends on the oxygen content of the surrounding atmosphere and the presence of magnesium.

Analysis of liquid metal foams through X-ray radioscopy and microgravity experiments.

Complex liquid structures such as metallic foams were produced in a furnace that allowed in situ X-ray monitoring of the evolution of the structure and distribution of the liquid in the foam. The experiments were carried out during parabolic flights which provided varying levels of gravity. The evolution of the characteristic liquid fraction profiles due to gravity induced drainage was measured and analysed in terms of the foam drainage equation, obtaining viscosity and surface tension by fitting solutions of the equation to the experimental data. The surface tension of the melt in the foam was decreased up to 40%. Effective viscosities of up to 139 times the viscosity of a pure bulk melt were observed. These effects could be attributed to the smaller influence of solid particles dispersed in the melt and the larger influence of the complex foam structure.

Inclusion complexation of phenoxyaliphatic acid derivatives of 3,3'-bis(indolyl)methanes with ß-cyclodextrin.

The inclusion complexation behavior of phenoxyaliphatic acid derivatives of 3,3'-bis(indolyl)methane (BIMs 1-5) with ß-cyclodextrin (ß-CD) were investigated in both solution and solid state by means of UV-Visible, fluorescence spectroscopy, FT-IR and (1)H NMR techniques. The nature of the host-guest inclusion complex between BIMs and ß-CD has been elucidated. The experimental results confirmed the existence of 1:1 inclusion complex of BIMs with ß-CD. The binding constants describing the extent of formation of the complexes have been determined using Benesi-Hildebrand plots using UV-Vis and fluorescence spectroscopy. BIMs exhibited an affinity for ß-CD. The spectral studies suggested the phenyl ring along with alkyl substitutions of BIMs is present inside of ß-CD cavity.

Sustainable biofuel synthesis from non-edible oils: a mesoporous ZSM-5/Ni/Pt catalyst approach.

This work examines the hydrodeoxygenation (HDO) activity of non-edible oils using a high surface area catalyst. The HDO activity was thoroughly examined and contrasted using the high surface area catalyst Ni/Pt-ZSM-5 as well as other supports like MCM-48 and H-beta. Ni/Pt bimetals supported on mesoporous ZSM-5 were created via reverse order impregnation to facilitate HDO of non-edible oils. Techniques such as XRD, FT-IR, BET, HR-TEM, HR-SEM, TPD, and TGA were used to characterize the produced catalysts. The synthesized catalysts considerably influenced the hydrodeoxygenation activities for the synthesis of lengthy chain hydrocarbons in a stainless-steel reactor with a high-pressure fixed bed between 300 and 375 °C under 10-40 bar hydrogen pressure. High levels of Ni/Pt-ZSM-5 acidity, textural, and H2 consumption qualities were discovered. Distributions of the products were also reviewed, along with comparisons of the structure-activity connections.

PEGylated dendrimer polystyrene support: synthesis, characterisation and evaluation of biologically active peptides.

Poly(N,N-bisethylamine) dendrimers with high content of poly(ethylene glycol) were synthesized on 3-(Acryloyloxy)-2-hydroxypropylmethacrylate-crosslinked polystyrene (PS-AHMA) resin and tested in various conditions of solid phase peptide synthesis. The dendritic templates were generated to the second generation on cross-linker active site of 3-(Acryloyloxy)-2-hydroxypropylmethacrylate (AHMA). First generation dendrimer was designed by series of four-stage reactions, such as Schiff base incorporation, acidolytic cleavage, diazotization and thionyl chloride treatment and same synthetic routes were followed for second generation also. Poly(ethylene glycol) (PEG1000) has been grafted to second-generation dendrimer and used to check various physico-chemical parameters in Fmoc/Boc peptide synthetic conditions. The utility of PEGylated dendrimer support was demonstrated by synthesizing biologically potent linear as well as disulfide-bonded peptide by Fmoc method.

Synthetic evaluation of disulphide-bonded sarafotoxin on a poly(oxy ether) grafted dendrimeric poly(alkyl amine) support for polymer assisted organic synthesis.

The present paper describes the synthesis, characterization and assessment of a novel class of insoluble polymeric polystyrene supports which combines polar poly(ethylene glycol)dimethacrylate as a cross-linker and poly(ethylene glycol) grafted poly(N,N-bisethylamine) as a dendritic template. Poly(N,N-bisethylamine) dendrimers were generated by a series of reactions such as Schiff base integration, acidolysis, diazotization and thionyl chloride treatment. The same successive sequences of reactions have been followed for second generation dendrimers also and subjected to PEGylation (PEG 600) to achieve the desirable physico-chemical properties. The applicability of the novel PEGylated dendrimer support was demonstrated by synthesizing linear as well as disulfide bonded peptides in high yields and purities.

Bioactive peptides laden nano and micro-sized particles enriched ECM inspired dressing for skin regeneration in diabetic wounds.

Hard to heal wounds such as diabetic wounds is one of the major problems in the healthcare sector. Delayed healing and shortfall of functional restoration at the wound site require upgraded wound management aids. In this study, we report that a nanofibrous mat enriched with bioactive peptides laden nano and microparticles achieve the requirements as an effective diabetic wound dressing. By means of electrospinning method, we fabricated Poly (lactic-co-glycolic acid)/Collagen nano-scale mat and surface functionalised with wound healing peptides, laden Chitosan nano and micro-sized particles, creating an Extracellular Matrix (ECM) -like structure with biomimetic features. The developed dressing displayed good cytocompatibility with Keratinocyte and fibroblast cells and enhanced their in-vitro cell proliferation and migration. Experiments in the streptozotocin-induced diabetic mice model showed that bioactive peptides released from Chitosan particles shorten the inflammatory stage and promote neovascularisation. The supporting nanoscale matrix promotes increased collagen deposition in the wound beds, thereby hastening the complete healing process by substantial tissue re-generation and functional restoration. The results evince that the nano/microparticles enriched nano-scale mat show potential as an effective wound repair dressing for diabetic wounds.

Dual growth factor entrapped nanoparticle enriched alginate wafer-based delivery system for suppurating wounds.

We have investigated the wound healing efficiency of calcium alginate wafer embedded with growth factor entrapped PLGA nanoparticle. Herein, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) entrapped PLGA nanoparticles were synthesized and embedded in a sodium alginate gel by freeze-drying technique. The synthesized dressing exhibited a high degree of swelling and appropriate porosity. The scaffold was characterized by Scanning Electron Microscopy (SEM) showing a highly porous morphology. Also, incorporation of growth factor loaded nanoparticles in a wafer-based delivery system resulted in localized growth factor delivery at the site of the wound in a sustained manner. The biocompatibility of the scaffold was evaluated by MTT assay, which showed a higher cell proliferation in the proposed scaffold as compared to the control. In vivo wound healing efficiency of the scaffold was evaluated using a full thickness murine wound model, which showed improved re-epithelialization, collagen deposition, and angiogenesis. These results suggest the use of the scaffold as a promising wound dressing material.

Sorafenib-Entrapped, Self-Assembled Pullulan-Stearic Acid Biopolymer-Derived Drug Delivery System to PLC/PRF/5 Hepatocellular Carcinoma Model.

Purpose: This study aimed to design a prototypic drug delivery system (DDS) made of an amphiphilic, pullulan (Pull)-derived biodegradable polymer for targeting the asialoglycoprotein receptor (ASGPR) overexpressed in HCC. Stearic acid (SA) was conjugated to increase the hydrophobicity of pullulan (Pull-SA). Methods: Pullulan (Pull) was linked to stearic acid (SA) after functional group modifications via EDC/NHS chemistry and characterized. Sorafenib tosylate (SRFT) was entrapped in pullulan-stearic acid nanoparticles (Pull-SA-SRFT) and its particle size, zeta potential, entrapment efficiency (EE), loading capacity (LC), and release efficiency was measured. The competence of Pull-SA-SRFT over SRFT in vitro was assessed using the ASGPR over-expressing PLC/PRF/5 hepatocellular carcinoma (HCC) cell line. This was done by studying cytotoxicity by MTT assay and chromosome condensation assay, early apoptosis by annexin-Pi staining, and late apoptosis by live-dead assay. The cellular uptake study was performed by incorporating coumarin-6 (C6) fluorophore in place of SRFT in Pull-SA conjugates. A biodistribution study was conducted in Swiss-albino mice to assess the biocompatibility and targeting properties of SRFT and Pull-SA-SRFT to the liver and other organs at 1, 6, 24, and 48 h. Results: The characterization studies of the copolymer confirmed the successful conjugation of Pull-SA. The self-assembled amphiphilic nanocarrier could proficiently entrap the hydrophobic drug SRFT to obtain an entrapment efficiency of 95.6% (Pull-SA-SRFT). Characterization of the synthesized nanoparticles exhibited highly desirable nanoparticle characteristics. In vitro, apoptotic studies urged that Pull-SA-SRFT nanoparticle was delivered more efficiently to HCC than SRFT. The cellular uptake study performed, gave propitious results in 4 hrs. The biodistribution study conducted in immunocompetent mice suggested that Pull-SA-SRFT was delivered more than SRFT to the liver when compared to other organs, and that the system was biocompatible. Conclusion: Pull-SA-SRFT is a promisingly safe, biodegradable, cell-specific nanocarrier and a potential candidate to target hydrophobic drugs to HCC.

"Shimming a Plate" Technique to Correct the Coronal Malalignment in Metadiaphyseal Distal Femur Fracture: Case Report.

CASE: We present a case of 19-year-old male patient with right sided comminuted metadiaphyseal distal femur fracture. Fixation was done using distal femur condylar locking plate. Washers were used with the screws to shimming the plate to achieve the coronal alignment and prevent stress over the screws. 2-year follow-up shows good alignment, radiological union and functional outcome without any complications. "Shimming" a locking plate with washers helps to achieve the better coronal alignment in comminuted metadiaphyseal distal femur fractures with coronal malalignment. It reduces the stress over screws and prevents implant-related complications.

Local delivery of hyaluronic acid as an adjunct to scaling and root planing in the treatment of chronic periodontitis in smokers and non-smokers: A clinical and microbiological study.

Objective: The objective of the study is to assess the effects of adjunctive use of hyaluronic acid (HA), clinically and microbiologically in smokers and nonsmokers, in the management of chronic periodontitis. Material and Methods: 48 sites from 24 chronic periodontitis patients, including smokers and nonsmokers with probing depth >5 mm, were selected for the study. A split-mouth design was followed. Scaling and root planing (SRP) was performed for all the patients. The test sites received 0.8% HA gel whereas the control site received placebo gel. The clinical parameters plaque, gingival and bleeding indices, Pocket Probing Depth (PPD), and clinical attachment level (CAL) were evaluated at baseline, 1 and 3 months. Microbiological parameters were evaluated at baseline and 1 month. Results: The results showed reduction in PPD and gain in CAL in both smokers and nonsmokers at the end of 3rd month. The improvements in the test sites were statistically significant when compared with that of control sites. The microbiological analysis showed a significant reduction in Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis at the test sites when compared to the controls in both the groups. Conclusion: The findings in both groups suggest that the use of locally delivered 0.8% HA gel may be valuable as an adjunct to SRP in the treatment of chronic periodontitis.