Progress in development of vaccine against Hyalomma anatolicum anatolicum-Indian scenario.
Hyalomma anatolicum anatolicum, a three host tick vector transmitting the causative agent of bovine tropical theileriosis, is widely distributed throughout India. As a component of integrated control measures against the tick vector, attempts have been made to identify candidate protein molecules for development of an anti-tick vaccine in the different stages of this tick species. By strategic methods of isolation of the targeted molecules using affinity purification of proteins showing reactivity with immunoglobulins of animals previously immunized with different sources of tick antigen, six proteins were isolated in a significantly pure form. The recovery percentage of the candidate proteins was very low in the range of 1.8-8.0%. The protective potentiality of the antigens was tested in immunization and challenge trials and maximum potential was observed in the proteins isolated from total larval extracts, nymphal extracts and in larval glycoprotein. One of the antigens with a molecular weight of 37kDa isolated from larvae of H. a. anatolicum was found to have some adverse effect on development of Theileria annulata in the vector tick. The progress in the development of immunoprophylactic measures against H. a. anatolicum is discussed.

title: Upcoming and future strategies of tick control: a review.
abstract: Ticks are distributed worldwide and significantly impact human and animal health. Due to severe problems associated with the continuous use of acaricides on animals, integrated tick management is recommended. Increasing public health concern over the tick-borne diseases demands the strategic control of ticks on animals that transmit diseases to human beings. Immunological control of tick vector of Kyasanur Forest Disease (KFD) on cattle and other wild reservoir hosts is one of the possible alternative strategy for reducing the transmission of KFD to man. Chemical-vaccine synergies have been demonstrated and a combination of chemical and vaccine for tick and tick-borne disease control has been identified as a sustainable option. Studies have suggested the possibility of vaccine strategies directed towards both tick control and transmission of pathogens. Besides tick vaccine, use of endosymbionts, which are essential for the survival of arthropod hosts, for the control of tick vectors will be one of the targeted areas of research in near future. India with huge natural resources of herbs and other medicinal plants, the possibilities of developing herbal acaricides is discussed. The future of research directed towards target identification is exciting because of new and emerging technologies for gene discovery and vaccine formulation.

title: In vitro and in vivo efficacy of Acorus calamus extract against Rhipicephalus (Boophilus) microplus
abstract: To develop a environment friendly control measure against cattle tick, Rhipicephalus (Boophilus) microplus, medicinally important plants were identified and extracts were prepared. Twelve 95% ethanolic, thirteen 50% hydroethanolic and nine hot water extracts were prepared and tested against laboratory reared homogenous colony of R. (B.) microplus. Amongst the 34 extracts, 26 extracts showed no mortality within 72 h of application while 12.0 ± 4.9% to 35.0 ± 9.6% mortality of treated ticks was recorded in other extracts. Of the effective extracts, the extract prepared from rhizome of Acorus calamus proved highly efficacious and 100% final mortality within 14 DPT was recorded. The LC85 value of the extract was determined as 11.26. In vivo experiments confirmed the efficacy of the extract up to 42%, and repeat application was required after 7 DPT. The extract was found safe and no reaction was observed when animals were treated with 50% of the concentration, which was five times of the concentration used for in vivo studies. The possibility of using the extract for the control of cattle tick is discussed.

title: Comparative immunopotentiating properties of saponin and incomplete Freund's adjuvant coupled to affinity purified larval antigen of Hyalomma anatolicum anatolicum.
abstract: Comparative immunostimulatory properties of saponin and incomplete Freund's adjuvant (IFA) were studied using affinity purified 39 kDa larval antigen of Hyalomma anatolicum anatolicum. Significant antibody response to 39 kDa antigen was detected in the sera of rabbits immunized with both 39 kDa plus saponin and 39 kDa plus IFA in comparison to their corresponding control animals. Insignificant differences were noted in the antibody response between the animals of two immunized groups. Upon challenge, the animals immunized with 39 kDa plus IFA rejected 76.2 +/- 9.7% of larvae and 45.8 +/- 4.1% of adults while in group of animals injected with 39 kDa plus saponin rejected 80.9 +/- 11.2% of larvae and 47.2 +/- 5.7% of adults.

title: Assessment of duration of immunity in crossbred cattle immunized with glycoproteins isolated from Hyalomma anatolicum anatolicum and Boophilus microplus
abstract: To develop immunoprophylactic measures against multi-tick infestation, two glycoproteins of 34 and 29 kDa were isolated from the larvae of Hyalomma anatolicum anatolicum and Boophilus microplus, respectively, and assessed for their efficacy against experimental challenge infestations. The synergistic effect of the antigens in the presence of incomplete Freund’s adjuvant was found to confer protection (DT%) in animals against 56.48% of larvae and 52% of adults of H. a. anatolicum, while the effect was 40% against adults of B. microplus. The efficacy (E%) of the antigens in combination against larvae and adults of H. a. anatolicum was calculated as 70% and 64.3%, respectively, and 63% against adults of B. microplus. A direct correlation between anti-glycoprotein antibody response and protection against infestation was observed. Western blot analysis detected specific antigen in the sera of animals of group A. The antigens in combination with incomplete Freund’s adjuvant could protect animals from H. a. anatolicum and B. microplus infestations for at least 30 weeks. The possibility of employing the vaccination strategy in Indian conditions is discussed.

title: Comparative diagnostic potentiality of ELISA and dot-ELISA in prepatent diagnosis of experimental Fasciola gigantica infection in cattle.
abstract: A glycoprotein (27 kDa) was isolated from crude somatic antigen of Fasciola gigantica by two steps affinity chromatography and was used in early detection of experimental fasciolosis in cattle by indirect ELISA and in dot-ELISA formats. Although, anti-27 kDa antibodies could be detected after 3 weeks post infection (WPI) by dot - ELISA which was one week later than indirect ELISA. The test, dot-ELISA, was more convenient in field application. By the test (dot-ELISA) the infection could be equally detected in animals infected with 100, 200 and 300 metacercariae of F. gigantica with high sensitivity. Further, the antigen (27 kDa) was not found to react with goat sera infected with Paramphistomum epiclitum, which are giving strong reaction to homologous immature and mature fluke antigens of P. epiclitum.

title: Comparison of the cortical connections of areas 4 gamma and 4 delta in the cat cerebral cortex.
abstract: Area 4 of the cat cerebral cortex has been subdivided into several regions based on cytoarchitectonic studies: areas 4 gamma, 4 delta, 4sfu, and 4fu (Hassler and Muhs-Clement [1964] J. Hirnforsch. 6:377-420). In a previous study, we found separate representations of contralateral limb movements in areas 4 gamma and 4 delta (Ghosh [1997] ). To investigate the relationship between these representations, the ipsilateral cortical connections of area 4 gamma and 4 delta were compared by the use of the retrograde neural tracers. After intracortical microstimulation of area 4, tracer was injected into one or two of the following regions: the forelimb regions of the rostral and caudal subdivisions of areas 4 gamma and 4 delta (r4 gamma, r4 delta, c4 gamma, c4 delta, separated by the cruciate sulcus) and the hindlimb regions of c4 gamma and c4 delta. Retrogradely labeled neural profiles were counted in every fourth section of the coronal series and located in cytoarchitectonic areas of the ipsilateral cortex. We found topographically organized reciprocal connections between areas 4 gamma and 4 delta; these connections were part of a rich network of interconnections between the cytoarchitectonic subdivisions of area 4. The forelimb regions of c4 gamma and c4 delta, of r4 gamma and c4 delta, and of r4 gamma and r4 delta were interconnected. These findings support the location of a secondary motor area in 4 delta. No interconnections between the forelimb regions of r4 gamma and c4 gamma, and of r4 delta and c4 gamma, could be found. Area 6 (particularly area 6a gamma) was found to project strongly to the forelimb regions of r4 gamma and r4 delta and relatively weakly to the forelimb region of c4 delta. Retrogradely labeled neurons were also detected in areas 3a, 3b, 1, 2pri, 5, 7, and insula after tracer injections in area 4.

title: The Sib TDT adjusted for age of disease onset.
abstract: Since the Sib-TDT (Spielman & Ewens, 1998) ignores age of disease onset, there could be loss of power in detecting linkage. In this article, we propose an adjustment for age of onset using the Cox Proportional Hazard Model (Cox, 1972) with the marker allele as a covariate. The test statistic for linkage is identical to the traditional Sib-TDT. Monte-Carlo simulations are performed under different disease models to assess the increase in power of the age-adjusted Sib-TDT compared to the traditional Sib-TDT. We extend our method to multiallelic markers. An application using data on Alzheimer's Disease is also presented.

title: Domain specific association of small fluorescent probe trans-3-(4-monomethylaminophenyl)-acrylonitrile (MMAPA) with bovine serum albumin (BSA) and its dissociation from protein binding sites by Ag nanoparticles: spectroscopic and molecular docking study.
abstract: Photoinduced intramolecular charge transfer produced a polar excited state in trans-3-(4-monomethylaminophenyl)acrylonitrile (MMAPA), rendering the resulting emission sensitive to the medium polarity. Strong binding interaction of silver nanoparticles with the probe was observed, causing fluorescence quenching through the static quenching process. The probe MMAPA was found to bind to the less polar hydrophobic, restricted proteinous environment of bovine serum albumin (BSA) resulting in the blue shift of the emission maximum with an increase in emission intensity and fluorescence anisotropy. Studies using site markers of flufenamic acid and phenylbutazone coupled with molecular docking results predicted that the binding site of the probe is in between subdomains IIIA and IB of BSA and is different from the conventional Sudlow sites. The denaturation of the probe-bound BSA by urea or heat released the probe from this proteinous environment to water marked by exactly reverse spectral changes. On the interaction of silver nanoparticles with the probe bound protein, the probe was observed to move from its binding site in the protein to the Ag(0) nanoparticle surface involving conformational changes of the protein near the probe binding site.

title: Analysis of the expression and function of Wnt-5a and Wnt-5b in developing and regenerating axolotl (Ambystoma mexicanum) limbs.
abstract: Urodele amphibians are unique adult vertebrates because they are able to regenerate body parts after amputation. Studies of urodele limb regeneration, the key model system for vertebrate regeneration, have led to an understanding of the origin of blastema cells and the importance of positional interactions between blastema cells in the control of growth and pattern formation. Progress is now being made in the identification of the signaling pathways that regulate dedifferentiation, blastema morphogenesis, growth and pattern formation. Members of the Wnt family of secreted proteins are expressed in developing and regenerating limbs, and have the potential to control growth, pattern formation and differentiation. We have studied the expression of two non-canonical Wnt genes, Wnt-5a and Wnt-5b. We report that they are expressed in equivalent patterns during limb development and limb regeneration in the axolotl (Ambystoma mexicanum), and during limb development in other tetrapods, implying conservation of function. Our analysis of the effects of ectopic Wnt-5a expression is consistent with the hypothesis that canonical Wnt signaling functions during the early stages of regeneration to control the dedifferentiation of stump cells giving rise to the regeneration-competent cells of the blastema.

title: Cloning of the p50 DNA binding subunit of NF-kappa B: homology to rel and dorsal.
abstract: The DNA binding subunit of the transcription factor NF-kappa B, p50, has been cloned. p50 appears to be synthesized as a larger protein that is then processed to its functional size. Sequence analysis reveals remarkable homology for over 300 amino acids at the amino-terminal end to the oncogene v-rel, its cellular homolog c-rel, and the Drosophila maternal effect gene dorsal. This establishes NF-kappa B as a member of the rel family of proteins, all of which display nuclear-cytosolic translocation. Protein sequence from the p65 polypeptide has established that it is not encoded in the same mRNA as p50. However, p65 appears homologous to c-rel, suggesting that c-rel may form heterodimers with p50 and rel may include a homodimerization motif.