Plasma Lipoproteins as Crucial Components of Host Defence Against Infections

Interactions between lipoproteins and infectious microorganisms are diverse and often multifaceted. There is a growing body of evidence which suggests that circulating plasma lipoproteins play an important role in warding off various infections. They are increasingly recognized as vital components of the host immune system. The purpose of this chapter is to provide the reader with an overview of this emerging domain. We review the anti-infective role of different lipoprotein particles and their components and further highlight the known molecular mechanisms involved therein. Instances where lipoproteins facilitate infections instead of protecting against them are also summarized. Finally, broad implications for the future in this active line of research are discussed

Detection of foot-and-mouth disease virus type O in recovered as well as healthy cattle to study carrier status in Assam

200-206Foot and mouth disease (FMD), one of the most contagious diseases of animals, affects different host species including wild animals. Asymptomatic FMD recovered animals may remain as carrier, which may be threat to other healthy animals. Hence, it is necessary to monitor the carrier status of the FMD recovered animals to effectively prevent further spread of the disease. Out of all the seven serotypes of FMD, O serotype is most commonly found in livestock. Therefore, in the present study, we chose to detect serotype ‘O’ in oropharyngeal fluid (OP) and to quantify cytokines, viz. IL-1α, IL-1β and IL-2. A total of 30 OP fluids and 30 blood samples were collected from 10 animals (1 in-contact healthy animal) for 3 months post infection. FMD O serotype could be detected in all the animals (100%). The RQ values were found to be 0.014 to 63.118 and 0.162 to 46.889 for IL-1α and IL-1β genes respectively, while insignificant RQ values were obtained for IL-2. In the second and third months, two animals showed down regulation for IL-1α gene, while IL-1β and IL-2 genes were down regulated in 7 animals and in all 10 animals, respectively for all the three months

Procjena orfvirusa (orfv) in vitro i in vivo elektronskom mikroskopijom

Transmission electron microscopy (TEM) was employed for describing skin and scab lesions in goats affected by orf virus and to demonstrate the parapoxvirus from clinical suspensions by negative staining and ORFV confirmation by immunogold electron microscopy. All samples were confirmed as parapoxvirus by semi-nested PCR amplification of partial gene encoding for the B2L envelope protein. Skin lesions were characterized by ballooning degeneration and loss of desmosomes of the spinosum cells, cytolysis and vesicle formation. Nuclear changes included chromatin margination and increase in electron density. Cytoplasmic changes were typical of cell swelling, vacuolation and the presence of uniform, moderately electron dense viroplasm, situated in the perinuclear region. Various intracellular forms including immature virions (IV), mature virions (IMV) and wrapped virions (WV) were observed in the cytoplasm. All these forms of ORFV were observed morphologically akin to vaccinia virus (VACV). Negative staining of clinical samples and viral suspensions showed typical parapoxvirus morphology with a characteristic criss-cross tubular surface pattern. Both ‘capsule’ form (‘C’ form) and ‘mulberry’ form (‘M’ form) of the virus were demonstrated. ORFV speciation was confirmed by immunogold tagging using a monoclonal antibody in ultra-thin skin section.U radu je za opisivanje lezija na koži i krastama kod koza inficiranih orfvirusom primijenjena transmisijska elektronska mikroskopija (TEM). Dokaz parapoksvirusa iz suspenzije i potvrda ORFV-a obavljeni su tehnikom negativnog bojenja te imunogold elektronskom mikroskopijom. Svi su uzorci potvrđeni kao parapoksvirusi pomoću poluugniježđene lančane reakcije polimerazom kojom je umnožen dio gena B2L odgovornog za protein ovojnice. Lezije na koži karakterizirane su balonskom degeneracijom i gubitkom dezmosoma spinoznih stanica, citolizom te stvaranjem vezikula. Promjene u jezgri uključivale su rub kromatina i povećanje elektronske gustoće. Citoplazmatske promjene bile su tipične za bubrenje stanica, vakuolaciju i prisutnost uniformnog, umjereno elektronički gustog viroplazma smještenog u međumembranskom području. U citoplazmi su zabilježeni različiti unutarstanični oblici, uključujući nezrele virione (IV), zrele virione (IMV) i omotane virione (WV). Svi su oblici ORFV-a pokazali morfološku srodnost s virusom vakcinije (VACV). Negativno bojenje uzoraka i suspenzija pokazali su tipičnu građu parapoksvirusa s karakterističnim križno-cjevastim uzorkom na površini. Također, prikazana su oba oblika virusa, kapsula (oblik C) i dud (oblik M). Specijacija ORFV-a potvrđena je imunogold obilježavanjem pri čemu su korištena monoklonska protutijela u ultratankom rezu kože

Association of Decreased High-Density Lipoprotein Cholesterol (HDL-C) With Obesity and Risk Estimates for Decreased HDL-C Attributable to Obesity

Background: Obesity is an important risk factor for decrease in high-density lipoprotein cholesterol (HDL-C) levels, which predisposes to cardiovascular diseases. But, the relative contribution of obesity toward decreased HDL-C and the risk estimates of decreased HDL-C attributable to obesity are unavailable. Such measures will help in understanding the extent by which the burden of decreased HDL-C can be reduced by tackling obesity. Objectives: The objectives of this study were to ( a ) determine the association between decreased HDL-C and obesity and ( b ) estimate the attributable risk proportion (ARP) and population attributable risk proportion (PARP) for decreased HDL-C due to obesity. Methods: Body mass index (BMI) and waist circumference (WC) were measured as indices of overweight (or generalized obesity) and central obesity, respectively in 190 subjects (95 cases with low HDL-C and 95 healthy controls with normal HDL-C) from Guwahati city. Crude odds ratio (OR) and adjusted OR with 95% confidence interval (CI) were calculated along with the risk estimates (ARP and PARP). Results: People with overweight or generlized obesity (adjusted OR = 4.90, 95% CI = 3.59-6.68), and people with central obesity (adjusted OR = 3.33, 95% CI = 2.39-4.64) had significantly greater odds of developing decreased HDL-C. Among the exposed, 79.8% of the decreased HDL-C cases could be attributed to overweight (or generalized obesity), while 72.8% cases could be attributed to central obesity. In the overall population, the corresponding figures were 57.1% and 36%, respectively. Conclusion: Decreased HDL-C is strongly associated with and largely attributable to obesity

A simplified multiplex PCR-based typing method for common Salmonella enterica serovars supported by online server-based detection system

Background & objectives: A rapid and simple alternative method is needed to replace the laborious, time-consuming Salmonella serotyping. The objective of the present study was to improve and simplify a previously reported multiplex polymerase chain reaction (PCR)-based method and to create an online server to enable rapid determination of serovars. Methods: A method of multiplex PCR-based genome typing (MPGT) was standardized using 59 Salmonella isolates of 31 serovars. Several previously reported primers were modified to obtain a more accurate performance. The screen was separated into four different multiplex reactions distinguishable on standard electrophoresis. A blind study was subsequently performed with 81 isolates of 10 serovars most prevalent in India. Whole genome information from 440 Salmonella isolates was used to confirm the usefulness of this method and concurrence of in silico predictions and PCR results were investigated. A public server (http://www.mpgt-salmonella.res.in) was established for data storage and determination of closest previously observed Salmonella isolates based on obtained MPGT patterns. Results: The 16 target genes amplified showed variability in their presence in strains from different serotypes. Hence, identical amplification patterns suggested genetic relatedness of strains and usually identical serological behaviour. The observed absence/presence patterns of genes were converted to an MPGT code. Altogether, 83 different codes were predicted in silico based on the whole genome information of 440 strains. Results confirmed that major serovars usually displayed unique MPGT codes. Interpretation & conclusions: The multiplex PCR assay resulted in specific binary codes for isolates from each of the 31 Salmonella serovars tested. The online server allowed the user to compare obtained PCR results with stored previous patterns. Simplicity, speed and cost-effectiveness make this tool useful for quick outbreak management

Molecular dynamics approach to probe the antigenicity of PagN – an outer membrane protein of <i>Salmonella</i> Typhi

<p>PagN is a highly immunogenic 27-kDa outer membrane adhesin present in <i>Salmonella</i> Typhi. It plays a major role in the pathogenesis of typhoid fever and has emerged as a strong vaccine candidate. In this report, we predict the three-dimensional structure of PagN and describe the conformational dynamics associated with its four extracellular loops based on two 100-ns molecular dynamics simulations at 300 and 310 K. The formation and deformation of the secondary structures on these loops were also investigated during the simulations which revealed loops L1 and L2 to be highly flexible, whereas the relative flexibility of loops L3 and L4 was minimal. Essential dynamics and principal component analysis deciphered more realistic dynamic behaviours of the loops, particularly at 310 K. Moreover, our epitope predictions suggest that the antigenic peptides for B-cell recognition are located within the loops L1 and L2, while those for T-cell recognition are located within the loops L3 and L4. The binding specificities of the antigenic peptides towards specific human MHC-I and MHC-II HLA alleles closely resembled the stability of the loops L3 and L4 inferred from the simulations. Finally, we identified potential antigenic peptides in the flexible (L1 and L2) as well as stable (L3 and L4) regions of PagN for both B- and T-cell recognitions, which can help in developing effective sub-unit vaccines.</p

Molekularna karakterizacija gena za mitohondrijsku 16S rRNA goveda, bivola i jaka.

A combination of polymerase chain reaction (PCR) with restriction fragment length polymorphism (RFLP) and nucleotide sequencing is the most preferred and efficient method for characterization of different species, in terms of detection power and applicability to large scale screening. The present study was carried out with the aim of developing the molecular fingerprint of the mitochondrial 16S rRNA gene of Cattle, Buffalo and Yak. Blood samples were collected randomly from ten different animals of each species for mitochondrial DNA extraction. The extracted DNA was used for the amplification of the 16S rRNA gene using universal primers. The size of the amplified products was 600bp. RFLP studies were carried out by digesting the amplicons using restriction enzymes viz. AluI, HinfI and HaeIII. The resulting RFLP pattern could easily identify and differentiate each of the species. Sequencing of the amplicons in all three species was carried out to confirm the variations at nucleotide level. Sequence analysis of the 16S rRNA gene using MEGA4 software and also PCRRFLP revealed that the 16S rRNA gene can be used as a good candidate for a molecular marker.Kombinacija lančane reakcije polimerazom (PCR) s polimorfizmom duljine restrikcijskih fragmenata (RFLP) i utvrđivanjem slijeda nukleotida (sekvenciranje) najpoželjniji je i učinkovit postupak za otkrivanje širokog raspona varijacija pri karakterizaciji različitih vrsta. Cilj ovog istraživanja bio je razviti molekularni otisak za gen koji određuje mitohondrijske 16S rRNA goveda, bivola i jaka. Za izdvajanje mitohondrijske DNA nasumično je prikupljeno po 10 uzoraka krvi različitih životinja unutar svake vrste. Izdvojena DNA iskorištena je za umnažanje 16S rRNA gena uz uporabu univerzalnih početnica. Veličina umnoženih proizvoda iznosila je 600 bp. RFLP analize provedene su digestijom amplikona pomoću restrikcijskih enzima viz. AluI, HinfI i HaeIII. Dobiveni RFLP uzorak mogao je lako prepoznati i razlikovati svaku od vrsta. Sekvenciranje amplikona u sve tri vrste provedeno je s ciljem da se potvrde varijacija na razini nukleotida. Analiza sekvencija pomoću računalnog programa MEGA4 i metoda PCR-RFLP pokazala je da se gen 16S rRNA može upotrijebiti kao dobar kandidat za molekularni biljeg

Detection of foot-and-mouth disease virus type O in recovered as well as healthy cattle to study carrier status in Assam

Foot and mouth disease (FMD), one of the most contagious diseases of animals, affects different host species including wild animals. Asymptomatic FMD recovered animals may remain as carrier, which may be threat to other healthy animals. Hence, it is necessary to monitor the carrier status of the FMD recovered animals to effectively prevent further spread of the disease. Out of all the seven serotypes of FMD, O serotype is most commonly found in livestock. Therefore, in the present study, we chose to detect serotype ‘O’ in oropharyngeal fluid (OP) and to quantify cytokines, viz. IL-1α, IL-1β and IL-2. A total of 30 OP fluids and 30 blood samples were collected from 10 animals (1 in-contact healthy animal) for 3 months post infection. FMD O serotype could be detected in all the animals (100%). The RQ values were found to be 0.014 to 63.118 and 0.162 to 46.889 for IL-1α and IL-1β genes respectively, while insignificant RQ values were obtained for IL-2. In the second and third months, two animals showed down regulation for IL-1α gene, while IL-1β and IL-2 genes were down regulated in 7 animals and in all 10 animals, respectively for all the three months

Evaluation of Pathogenicity and Structural Alterations for the Mutations Identified in the Conserved Region of the C‑Terminal Kinase Domain of Human-Ribosomal S6 Kinase 1

Human-ribosomal s6 kinase 1 (h-RSK1) is an effector kinase of the Ras/MAPK signaling pathway, which is involved in the regulation of the cell cycle, proliferation, and survival. RSKs comprise two functionally distinct kinase domains at the N-terminal (NTKD) and C-terminal (CTKD) separated by a linker region. The mutations in RSK1 may have the potential to provide an extra benefit to the cancer cell to proliferate, migrate, and survive. The present study focuses on evaluating the structural basis for the missense mutations identified at the C-terminal kinase domain of human-RSK1. A total of 139 mutations reported on RSK1 were retrieved from cBioPortal, where 62 were located at the CTKD region. Furthermore, 10 missense mutations Arg434Pro, Thr701Met, Ala704Thr, Arg725Trp, Arg726Gln, His533Asn, Pro613Leu, Ser720Cys, Arg725Gln, and Ser732Phe were predicted to be deleterious using in silico tools. To our observation, these mutations are located in the evolutionarily conserved region of RSK1 and shown to alter the inter- and intramolecular interactions and also the conformational stability of RSK1-CTKD. The molecular dynamics (MD) simulation study further revealed that the five mutations Arg434Pro, Thr701Met, Ala704Thr, Arg725Trp, and Arg726Gln showed maximum structural alterations in RSK1-CTKD. Thus, based on the in silico and MD simulation analysis, it can be concluded that the reported mutations may serve as potential candidates for further functional studies