Occurrence of Chromosomal Polymorphism in a Striped Ichthyophid Caecilian (Amphibia: Gymnophiona) from Western Ghats of India

Mitotic and meiotic chromosomes prepared from specimens of Ichthyophis italics tricolor collected from either side of Palghat Gap from three different locations of Western Ghats revealed a diploid number (2n=42-44) with Fundamental Number (62-64). Highest diplod number (44) and occurrence of chromosomal polymorphism is documented for the first time for gymnophion amphibians. Role of chromosomal rearrangements involved in karyotypic derivation process is discussed

Karyological Characteristics of Two Species of Unstriped Ichthyophid Caecilians (Amphibia:Gymnophiona:Ichthyophiidae) of Western Ghats of India

Karyotypic details of two species of unicoloured caecilians namely Ichthyophis malabarensis (2n=42-44, FN=60,66) and Ichthyophis peninsularis (2n=42, FN=58) collected from Western Ghats of India, is reported for the first time. Role of chromosomal rearrangements involved in karyotypic derivation is discussed. The present chromosomal data suggest that there exists more than one species of unstriped ichthyophid caecilian species in Western Ghats of Indi

Purification and characterisation of a carboxylesterase from the latex ofSynadenium grantii Hook, ‘f’

The latex ofSynadenium grantii was found to contain esterolytic activity. Polyacrylamide gel electrophoretic study coupled with substrate and inhibitor specificity studies revealed the presence of multiple forms of carboxylesterases and cholinesterases in the latex. One of the carboxylesterases of the latex was purified by acetone fractionation, carboxymethyl-Sephadex chromatography and Sepharose-6B gel filtration. The homogeneity of the enzyme was established by polyacrylamide gel electrophoresis, isoelectric focussing and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The enzyme consists of a single polypeptide chain with a molecular weight of 14,000. The amino acid analysis of the purified enzyme revealed that it contained a greater number of neutral and acidic, compared to basic amino acid residues. The isoelectric pH of the enzyme was found to be 4.0. The enzyme was a glycoprotein as revealed by periodic acid Schiff-staining technique. Studies with different organophosphate and carbamate inhibitors showed that this enzyme was sensitive to organophosphates. The product inhibition studies with this enzyme showed linear competitive inhibition with acetate and linear non-competitive inhibition with 1-naphthol

Kvercetin potpomaže akaricidnu aktivnost ivermektina u slučaju svinja prirodno oboljelih od sarkoptoze

Sarcoptic mange triggers continuous oxidative onslaughts, resulting in severe oxidative stress in pigs and, to date, no antioxidant has been evaluated for the treatment of naturally infested pigs. This randomized clinical trial aimed to assess the ameliorative potential of the antioxidant quercetin (QR) when integrated with ivermectin (IVM) in the treatment of sarcoptic mange in pigs. The control group (T0 , n=10) consisted of healthy subjects. The first treatment group (T1 , n=10) consisted of infested pigs receiving the standard treatment (subcutaneous IVM only) while the second treatment group (T2 , n=10) consisted of infested pigs receiving integrated treatment (subcutaneous IVM plus oral QR). On day 0, the circulating malondialdehyde (MDA) was significantly higher and superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), total antioxidant capacity (TAC) and antioxidative minerals (zinc, copper, iron) were lower in all infested pigs compared to the healthy subjects. On day 14 post-treatment, maximum recovery was observed in the MDA, SOD, GSH, CAT, TAC, zinc, copper and iron in group T2 and the results returned to normal earlier in group T2 than in T1 . Likewise, more significant improvements in parasitological cure rate, scratching index and skin score were recorded after treatment in group T2 than group T1 . These results suggest the greater effectiveness of IVM plus QR than IVM alone against sarcoptic mange, and quercetin may be recommended as an ancillary therapy with IVM to negate severe oxidative stress, improve post-therapy convalescence and produce a speedy recovery in pigs.Sarkoptoza u svinja pokreće poremećaje koji rezultiraju teškim oksidacijskim stresom za koji još uvijek nije otkriven antioksidans kojim bi se prirodno infestirane svinje tretirale. Cilj je ovog randomiziranog kliničkog istraživanja bio procijeniti antioksidacijski potencijal kvercetina (QR) u kombinaciji s ivermektinom (IVM) u liječenju sarkoptoze u svinja. U kontrolnoj su skupini (T0 , n=10) bile zdrave jedinke. U prvoj su pokusnoj skupini (T1 , n = 10) infestirane svinje dobile standardnu terapiju (samo IVM primijenjen supkutano), dok su infestirane svinje u drugoj pokusnoj skupini (T2 , n = 10) primile integriranu terapiju (supkutano IVM i oralno QR). Nulti dan cirkulacijski je malondialdehid (MDA) bio znakovito veći, dok su superoksidna dismutaza (SOD), reducirani glutation (GSH), katalaza (CAT), ukupan anitoksidacijski kapacitet (TAC) i antioksidacijski minerali (cink, bakar i željezo) bili smanjeni u infestiranih svinja u usporedbi sa zdravim jedinkama. Četrnaesti dan poslije liječenja uočen je maksimalan oporavak u pogledu pokazatelja MDA, SOD, GSH, CAT, TAC, cinka, bakra i željeza u skupini T2 te njihov raniji povratak na uobičajene vrijednosti u skupini T2 u odnosu na skupinu T1. Osim toga, u skupini T2 u odnosu na skupinu T1 zapaženo je znakovito poboljšanje u stopi izliječenosti parazitoze, indeksu grebenja i bodovanju promjena na koži. Ovi rezultati upućuju na veću učinkovitost IVM-a u kombinaciji s QR-om nego IVM-a upotrijebljenog kao samostalna terapija sarkoptoze u svinja. Zaključuje se da bi kvercetin mogao biti dodatna terapija uz IVM kako bi se poništili teški učinci oksidacijskog stresa, poboljšala poslijeterapijska rekonvalescencija i ubrzao oporavak svinja

Immunopharmacological consequences of immune responses to therapeutic interferon beta

Protein therapeutics or biologics represent 30 % of current licensed pharmaceutical products. In general, biologics offer superior safety profiles compared to small molecules. However, significant clinical concerns have emerged in terms of development of anti-drug antibodies (ADAs), a phenomenon that is covered under the term, immunogenicity. Anti-drug antibodies can alter pharmacokinetics, reduce efficacy of the therapeutic and also can in some cases induce allergic reactions. Human recombinant interferon beta (IFN-β) is a biologic used for the treatment of multiple sclerosis (MS) – a chronic, inflammatory and demyelinating disease of the central nervous system. Long-term treatment with IFN-β has been shown to lead to the development of anti-IFN-β antibodies that can cause total loss or reduced efficacy. Anti-drug antibodies can be non-neutralising (N-NAb) and neutralising (NAb) depending on the site to which they bind. This study aimed to conduct a systematic review to determine factors affecting the formation of neutralising antibodies against three different formulations of IFN-β Avonex™, Rebif™ and Betaseron/Betaferon™. Findings from the systematic review highlight the relative differences in immunogenicity risk of different IFN-β formulations Avonex™, Rebif ™ and Betaseron/Betaferon™ with Avonex™ having the lowest risk, Rebif™ has moderate risk and Betaseron/Betaferon™ has high risk. Characterising the immunoglobulin profile of the IFN-β ADAs from the plasma of ADA positive MS patients revealed that IFN-β ADAs are predominantly of the IgG1 and IgG4 subclass. We also characterised the neutralising potential of the major ADA IgG4 subclass using a IFN-β bioactivity assay and show that IgG4 antibodies may likely contribute to the neutralisation activity. The potential of the neutralising ADAs to cross-react with endogenous IFN-β was investigated using an in vitro bioactivity assay. Findings from this set of experiments revealed varying degrees of neutralisation of endogenous IFN-β. We next explored the potential immunological consequence of ADA with regards to formation of immune complexes and activation of complement. The interaction of ADAs with the biologic can result in formation of immune complex. Immune complexes can activate the complement system. The data revealed IFN-β ADAs can form immune complexes with IFN-β and therefore activate complement. We also attempted to identify IFN-β linear epitopes that the ADAs had the ability to bind. However, a combination of multipin peptide technology and in vitro peptide competitive binding assay failed to reveal a definitive linear epitope although there was some evidence for the existence of potential linear epitopes. We also examined the involvement of T helper cells and T regulatory cells (Tregs) in ADA development. The data revealed no significant differences in the frequency of Tregs among IFN-β ADAs positive, negative and healthy donors. Attempts were also made to identify T helper epitopes within IFN-β that could potentially drive immunogenicity. Using T-cell epitope prediction tools (IEDB-AR and ProPred) and T-cell functional assays we identified an immunogenic sequence of 36 amino acids within IFN-β (position 130-166). Our data revealed that one IFN-β peptide within this sequence is a potential T-cell immunogenic epitope. In addition we identified a possible association of one IFN-β derived peptide with the DRB1*1501HLA haplotype. In summary, the results presented in this thesis have provided essential information on subclass profile of IFN-β ADAs, the possible involvement of T helper cells and potential antibody epitopes within IFN-β. Future studies should be aimed at providing greater detail on the evolution of the ADA response and test strategies to remove immunogenic determinants from IFN-β

Global positioning system based spatial and temporal distribution of new leaf curl begomovirus disease on sunflower in Northern Karnataka

Leaf curl disease on sunflower caused by begomovirus genus of the family geminiviridae. Present investigations on field survey for disease incidence, field diagnostic symptoms and its spatial and temporal distribution in major sunflower growing parts of North Eastern Karnataka through GPS system during 2013-14, revealed that the disease was found to occur at all the stages of sunflower under field condition and exhibited symptoms such as vein thickening (enations) on abaxial surface of the leaves, upward curling and reduction in leaf size and severe discoluration of capitulum (Head) followed by bushy appearance. GPS based survey indicated that the % disease incidence varied from location to location (spatial variation) and also from season to season (temporal variation). The low incidence was noticed during Kharif condition which is ranged between 6.34-11.16, with the average incidence of 11.2%, 7.4% and 6.3% in Koppal, Raichur and Ballari districts repectively. Whereas during Rabi/summer season, high magnitude of disease noticed in many of the locations surveyed and is recorded upto 92.9 %. The GPS maps plotted based on PDI scale (0-3) represents high risk areas of the disease in Raichur and adjacent areas of Nort Eastern Karnataka and the result shows that the disease occurrence was more in rabi as compared to Kharif situations irrespective of locations. GPS survey map is an indicator to locate the nature of disease spread so as to conclude the hotspot areas

Biological and molecular evidences on host range of leaf curl begomovirus disease of sunflower (Helianthus annuus L.)

The present study was conducted to identify the alternate hosts of new leaf curl virus disease of sunflower. In the present study several crops and weed hosts were cross inoculated with leaf curl virus of sunflower under laboratory through insect vector whitefly (Bemisia tabaci), further all inoculated samples were retested (3-4 weeks after inoculation) by molecular based Polymerse chain reaction diagnosis for the presence of virus. The results revealed that the causal virus of the disease was successfully transmitted from sunflower to sunflower (Helianthus annuus), tomato (Solanum lycopersicum) and tobacco (Nicotiana tabacum L) and weed hosts such as Acanthospermum hispidum, Amaranthus viridis and Parthenium hysterophorus in a short incubation period (2-3 weeks after inoculation), while on other hosts Chilli (Capsicum annuum L) and Datura stramonium, infection occurs in delayed incubation period. Further molecular analysis thorough polymerase chain reaction (PCR) diagnostic technique using virus specific primers also confirmed the presence of coat protein (CP) of leaf curl begomovirus invirus inoculated hosts viz., chilli, sunflower, tomato, and tobacco and weed hosts such as Acanthospermum hispidum, Amaranthus viridis, Datura stramonium and Parthenium hysterophorus. Thus, findings substantiate that the above hosts are major sources of the virus inoculum and served as potential alternate hosts of the disease during the off season

Antimicrobial, Antioxidant Activity and Phytochemical Screening of Tecoma stans (L.) Juss. ex Kunth.

The ethanol, methanol and water extracts of Tecoma stans effective against tested bacteria (Pseudomonas fluorescens, Clavibacter michiganensis sub sp. michiganensis, Xanthomonas axanopodis pv. malvacearum, Staphylococcus aureus, E. coli, Pseudomonas aeruginosa and Klebsiella pneumonia) and fungi (all species of Aspergillus and Alternaria). Phytochemical analysis revealed the presence of alkaloids, flavonoids, saponins, phenols, steroids, anthraquinones and tannins. The three extract fractions have showed highest total Phenolic content (177-216 mg gallic acid equivalent/g). These three solvent fractions possessed strong radical scavenging activity from FRAP and DPPH. It was ranged from 1433.75 to 3841.17 g/ml. The results indicate that this plant is a potential candidate to be used as an antimicrobial and antioxidant.Key words: Tecoma stans, Antimicrobial, antioxidant, phytochemicals Govindappa M et al. Antimicrobial, Antioxidant Activity and Phytochemical Screening of Tecoma stans (L.) Juss. ex Kunth. J Phytol 3/3 (2011) 68-76