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This communication reports about heat shock protein response after arsenic exposure in broiler chickens in vivo and in vitro both. Splenocytes harvested in presence of sodium arsenite expressed Heat shock protein 70 (HSP 70) which could be identified on the basis of relative migration pattern and western blot analysis. Serum levels of HSP 70 in broiler chicken also increased after continuous feeding of sodium arsenite in drinking water. This particular observation may be attributed towards systematic inflammation, oxidative stress and hepatocellular injury. In vitro relative quantification of transcription level of HSP 70 revealed that splenocytes harvested in presence of sodium arsenite expressed (final concentration 3 and 7 μM/ml) more HSP 70 in comparison to cells harvested without sodium arsenite and the values were statistically significant ( P < 0.001) when compared to untreated control. Collectively this result indicated that, HSP 70 level increased both in vivo and in vitro trials and may be used as potential molecular and toxicological biomarker. [ABSTRACT FROM AUTHOR]

Comparison of nucleotide sequences at the VP1 coding region of foot-and-mouth disease serotype Asia1 viruses from India has revealed two genetic lineages with emergence of a genetically divergent group in recent years. In this study a simple, fast, relatively costeffective multi-primer RT-PCR assay to differentiate genetic lineages of type Asia1 viruses was developed. Efforts were made in the design of novel lineage-specific primers and in optimization of the multi-primer assay protocol in conjunction with the use of the serotype specific primer for confirmation of serotype Asia1 virus. This assay promises to be an effective tool in molecular epidemiological investigation of FMD in the country. [ABSTRACT FROM AUTHOR]

The amino acid composition of the two surface proteins of peste-des-petits ruminants vaccine virus belonging to lineage four from India were deduced from the nucleotide sequence. The fusion (F) protein gene of PPRV Sungri/96 is 2405 nucleotides long and in relation to the length, it is 80 nucleotides longer than that of PPRV Nigeria/75/1 which are found to be present at the 5′UTR of this virus. The complete F gene alignment with other morbillivirus reveals a homology of 89% with PPRV/Nigeria/75/1 and 48–51% with other morbilliviruses. The F protein of PPRV Sungri/96 exhibited characteristics similarity to those of other morbillivirus F proteins. The overall amino acid similarity with its counterpart PPRV Nigeria/75/1 was 96%; with other morbilliviruses it is 65–74%. The PPRV Sungri/96 haemagglutinin (H) protein gene is 1954 nucleotides long and showed a sequence homology of 90.7% with PPRV/Nigeria/75/1 and with other morbilliviruses it ranged from 33% to 45%. At amino acids level, PPRV Sungri/96 showed a homology of 92.3% with PPRV/Nigeria/75/1 and 34–49% with other morbilliviruses. The phylogenetic tree constructed for F and H gene reveals four separate groups which is very similar to that found in other genes. To the best of our knowledge this is the first report describing the F and H genes of an Indian isolate. [ABSTRACT FROM AUTHOR]

The complete nucleotide sequence of the large polymerase (L) protein of the peste-des-petits ruminants (PPR) vaccine virus (PPRV Sungri/96) belonging to the Asian lineage was determined. The gene was 6643 nucleotides in length from the gene-start to the gene-end and encoded a polypeptide of 2183 amino acids. The PPRV Sungri/96 has a nucleotide homology of 94.1% for PPRV Nigeria 75/1 to 64.4% for Canine distemper virus. At amino acid level PPRV Sungri/96 has an amino acid identity of 96.2% with PPRV Nigeria 75/1 and 70.4% to 74.8% with other morbilliviruses. All the established domains in L protein characteristic of paramyxoviruses were also found to be present in PPRV Sungri/96. Phylogenetic analysis of different L proteins of morbilliviruses revealed five well-defined clusters as observed previously. The 3′ trailer sequence of PPRV Sungri/96 is of 37 nucleotides long which is very similar to that of other morbilliviruses. To the best of our knowledge this is the first report describing the polymerase gene sequence of PPRV Indian isolate. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease (FMD), the most economically important disease of cloven-hoofed animals, is endemic in India. Sequence analysis revealed that phylogenetic grouping of type Asia1 field isolates on the basis of the large fragment of the 5′untranslated region (5′LF-UTR) was quite similar to that based on the sequences of the capsid-coding (VP1) region of the same viruses. The existence of two distinct lineages of type Asia1 suggested by the study on the VP1 region was further supported by the detection of a difference in length and predicted secondary structure of the 5′LF-UTR between the two lineages. Sequence variability between the isolates of the two lineages was also observed within the different domains of the internal ribosome entry site (IRES) around conserved motifs like the GNRA,- RAAA,- and the polypyrimidine tract. Certain group and lineage-specific signature nucleotides pertaining to FMDV type Asia1 in the 5′LF-UTR have been identified. The present study shows that the 5′LF-UTR of FMDV serotype Asia1 field isolates are variable in relation to the length and probable secondary structure of the IRES. [ABSTRACT FROM AUTHOR]

The capsid-coding (P1) and 3A regions of foot-and-mouth disease virus (FMDV) type A field isolates including two vaccine strains collected during 1977-2000 were analyzed. In the phylogenies, the isolates were distributed into two previously identified genotypes VI and VII, with multiple sub-genotypes that are temporally clustered. Comparison of the antigenic relationships of field isolates with the two vaccine strains (IND 17/77 and IND 490/97) and the reference strains of the genotypes VI (IND 233/99) and VII (IND 40/00) indicated two broad antigenic groups that correlate with the phylogenetic groupings (genotypes VI and VII), and are highly divergent from the vaccine strains. The maximum likelihood method of selection analysis identified a number of amino acid sites within the P1 region to be under weak positive selection. Some of the positively selected sites were mapped at/near the antigenically critical amino acid sites of the P1 region, indicating host immune pressure as one of the important driving force behind the observed genetic and antigenic diversity in FMDV. A small number of selected sites are located in the heparan sulphate-binding pocket of the virus, suggesting a fitness advantage for cell entry of the virus. No positive selection was detected in the 3A dataset. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease viruses (FMDV) have a high rate of mutation and spontaneous mutants can be readily isolated in the laboratory. In this study, plaque purified FMDV Asia1 vaccine strains (IND 63/72 and IND 491/97) were passagedin-vitroin Baby Hamster Kidney-21 cell monolayers in the presence of sub-neutralizing levels of antiviral polyclonal sera (APS), raised in guinea pigs against the purified and inactivated whole virus particles of IND 63/72, IND 491/97 and IND 13/01. After serial passages under selective immune pressure, the viruses starts growing in the presence of undiluted sera and showed certain characteristics like an increased resistance to neutralization by APS and reduction in plaque counts on titration in plaque assay. Cross-neutralization of these viruses with above-mentioned APS revealed selection of three complete and one partial polyclonal antibody resistant (PAR) viruses based on the ‘r’ value in micro neutralization test. Alterations were detected at several amino acid residues in the structural protein-coding P1 region. Many of the residues inferred to be positively selected sites in other serotypes of this virus were also prone to substitution under immune selection pressure in Asia1 virus. The present work extends the finding that selection exerted by host antibody also plays a major role in the rapid evolution of FMDV Asia1, as observed in other serotypes. [ABSTRACT FROM AUTHOR]

Summary Changes in the nucleotide sequence of the 1D gene of two vaccine strains (IND 63/72 and IND 491/97) of Foot-and-mouth disease virus (FMDV) serotype Asia1 during serial cytolytic infections in cell culture have been analyzed. Sequence comparisons revealed a majority of transition mutations in IND 491/97. The mutation frequency of the 1D gene of IND 491/97 was about 4.5 to 6.0 fold higher than that of IND 63/72. At the amino acids 40–60 and 140–160 regions the mutation frequency was higher compared to the whole VP1. Both viruses showed a constant change at certain residues of the G-H loop region with an accumulation of amino acid replacements during serial cytolytic passages in cell culture. The critical residues (145 and 153) identified previously using mAbs recognizing trypsin-sensitive epitopes were not substituted in the absence of immune selection but changes were observed at positions 142 and 148. Non-reactivity of IND 63/72 after 50th passage level onwards with a panel of mAbs indicated an alteration in the antigenic specificity of the virus. Comparison of amino acid sequences in the entire capsid coding region of the naturally occurring field isolates with that of the 50th and 100th passage level viruses of IND 63/72 revealed that the residues 56 and 74 of VP2 could be involved in mAb binding. The results suggest that fixation of amino acid replacements occurs in VP1 of Asia1 virus, which could play an important role in antigenic variation by modulating different antigenic epitopes located on the surface of the virus. [ABSTRACT FROM AUTHOR]

Summary. We have analyzed isolates of two recently circulating genotypes (genotypes VI and VII) of type A foot-and-mouth disease virus (FMDV) from India. Maximum-likelihood models provided support for the presence of positively selected sites in the capsid-coding (P1) region. Positive selection was detected at a number of amino acid positions behind a background of strong purifying selection. Among the positively selected sites, four were identified at known critical antigenic residues (VP2 79, VP3 59 and 70 and, VP1 83), suggesting that FMDVs are under pressure from the immune system. Two residues (VP2 134 and VP3 59) that are part of the heparan sulfate-binding pocket in subtype A22 FMDV are also inferred to be under positive selection. Antigenic divergence was observed between and within the genotypes in neutralization tests with sera raised against the representative isolates from genotypes VI and VII. The two vaccine strains showed one-way antigenic relationships (r value) of <0.2 with 64% of the isolates, whereas, with genotypes VI and VII an r value of >0.4 was observed with 24% and 64% of the isolates, respectively. No correlation could be deduced from the amino acid substitutions at specific critical residues and lower r values in the field isolates. [ABSTRACT FROM AUTHOR]

Cyclophosphamide (Cy) is widely used as an effective cytotoxic drug, but its use is limited because of its toxicity. In this report, we describe for the first time the ability of purified protein A (P) of Staphylococcus aureus to reduce Cy-induced toxicity in rats. Protein A-treated animals recover quickly from the toxic effects of Cy. The antitumor property of Cy is not reduced in the P + Cy group. In fact, the latter showed a persistent decrease in tumor volume compared with the Cy group. Protein A may prove to be an effective agent in increasing the therapeutic index of Cy. [ABSTRACT FROM AUTHOR]

Background: Accessory breast tissue can be present anywhere in the milk ridge because of incomplete involution. However, lactating accessory breast in the groin is rare. Methods: Case report. Results: We report a 36-year-old para 2, gravid 2, woman presenting with lactating accessory breast in the groin. She had similar symptoms during breastfeeding her first child but the symptoms subsided after she stopped breastfeeding only to recur during nursing her second child born 5 years after. Biochemical testing of the secretion confirmed it as milk. Aspiration cytology by fine needle of the swelling confirmed the breast tissue. She had no urogenital anomaly. Excision of the accessory tissue gave her relief. Conclusions: Awareness of this rare but interesting abnormality in embryological development is important. One should look for the associated abnormalities. If conservative measures fail, excision is the treatment of choice. [ABSTRACT FROM AUTHOR]

Plastic flow behaviour of low carbon steel has been studied at room temperature during tensile deformation by varying the initial strain rate of 3.3 x 10-4 s-1 to a final strain rate ranging from 1.33 x 10-3 s-1 to 2 x 10-3 s-1 at a fixed engineering strain of 12%. Haasen plot revealed that the mobile dislocation density remained almost invariant at the juncture where there was a sudden increase in stress with a change in strain rate and the plastic flow was solely dependent on the velocity of mobile dislocations. In that critical regime, the variation of stress with time was fitted with a Boltzmann type Sigmoid function. The increase in stress was found to increase with final strain rate and the time elapsed in attaining these stress values showed a decreasing trend. Both of these parameters saturated asymptotically at a higher final strain rate. [ABSTRACT FROM AUTHOR]