Werner protein is a target of DNA-dependent protein kinase in vivo and in vitro, and its catalytic activities are regulated by phosphorylation

Human Werner Syndrome is characterized by early onset of aging, elevated chromosomal instability, and a high incidence of cancer. Werner protein (WRN) is a member of the recQ gene family, but unlike other members of the recQ family, it contains a unique 3'-->5' exonuclease activity. We have reported previously that human Ku heterodimer interacts physically with WRN and functionally stimulates WRN exonuclease activity. Because Ku and DNA-PKcs, the catalytic subunit of DNA-dependent protein kinase (DNA-PK), form a complex at DNA ends, we have now explored the possibility of functional modulation of WRN exonuclease activity by DNA-PK. We find that although DNA-PKcs alone does not affect the WRN exonuclease activity, the additional presence of Ku mediates a marked inhibition of it. The inhibition of WRN exonuclease by DNA-PKcs requires the kinase activity of DNA-PKcs. WRN is a target for DNA-PKcs phosphorylation, and this phosphorylation requires the presence of Ku. We also find that treatment of recombinant WRN with a Ser/Thr phosphatase enhances WRN exonuclease and helicase activities and that WRN catalytic activity can be inhibited by rephosphorylation of WRN with DNA-PK. Thus, the level of phosphorylation of WRN appears to regulate its catalytic activities. WRN forms a complex, both in vitro and in vivo, with DNA-PKC. WRN is phosphorylated in vivo after treatment of cells with DNA-damaging agents in a pathway that requires DNA-PKcs. Thus, WRN protein is a target for DNA-PK phosphorylation in vitro and in vivo, and this phosphorylation may be a way of regulating its different catalytic activities, possibly in the repair of DNA dsb.     

Quantitative digital and palmar dermatoglyphics: Sexual dimorphism in the Chuvashian population of Russia

With the aim of determining sex dimorphism among the Chuvashian population of Russia, digital and palmar dermatoglyphics of 547 individuals (293 males, 254 females) were analyzed. The sex differences for PII, TRC, and AFRC are similar to Indian and Jewish populations. Correlation coefficients between individual finger ridge counts are a little lower than in Jews but are almost equal to Indian populations. The Mantel test of matrix correlation between sexes for 22 traits shows a very good similarity. However, sex differences of palmar traits display different levels when compared with other human populations. In light of this, our evidence indicates the possible role of environmental (prenatal) factors in the realization of dermatoglyphic sex differences. The development of palmar dermatoglyphics has had a relatively longer growth period compared with fingers [Cummins, H., 1929. The topographic history of the volar pads (walking pads, tast ballen) in the human embryo. Embryol. 20, 103-126]. The palmar dermatoglyphic pattern of affinities therefore corresponds better than fingers to the ethno historical background of the populations, ascertained by numerous studies.     

Dermatoglyphic sexual dimorphism: finger and palmar qualitative characteristics in five endogamous populations of West Bengal,India

Five hundred families from five different endogamous populations encompassing the main social rank in the caste hierarchy of the same geographical area of West Bengal, India, were analyzed to present variation in qualitative pattern types on fingers and palms. Sex dimorphism, homogeneous in all populations, suggests common characteristics of dermatoglyphic patterns. The pattern types are not uniformly distributed on 10 fingers and palmar configurational areas. However, most of these observations are homogeneous in nature, in both sexes among 5 populations. But the two sets of results on fingers and palms are not exactly the same. Palmar dermatoglyphic relationship reflects the better caste affinities, perhaps due to embryological development, having relatively a longer growth period compared to fingers (Cummins 1929). The present findings indicate that the qualitative dermatoglyphic affinities conform to the known ethnohistorical background of these populations, which correspond also to the results of quantitative dermatoglyphics as well as serological and biochemical markers of these populations. These observations indicate that these population groups have a common genetic background and thus traditional grouping of Indian populations on the basis of caste hierarchy may not be a reflection of the genetic origin of the population. In dermatoglyphic affinities, both qualitative and quantitative traits therefore may be quite useful in tracing the ethnohistorical background of these populations.     

Genetic determination of head-size-related anthropometric traits in an ethnically homogeneous sample of 373 Indian pedigrees of West Bengal

The substantial involvement of genetic factors in the determination of head-size and head-shape traits has been firmly established. However, there has been a lack of agreement on a number of specific issues concerning the pattern of inheritance of craniofacial features. In this study we examined some of these issues in a large, ethnically homogeneous sample of Indian pedigrees. The data included 1,263 individuals belonging to 373 nuclear families. Eleven raw head-size traits and two synthetic phenotypes, interpreted as horizontal and vertical head-size components (HOC and VEC, respectively), were used in the analysis. To establish the pattern of inheritance of head traits, we carried out univariate and bivariate analyses. Maximum heritability estimates ranged from 0.41 to 0.83 for the studied head-size phenotypes. The portion of the total residual variance attributable to putative additive genetic factors was 68.3% and 70.3% for HOC and VEC, respectively, and common familial factor effects were found to be nonsignificant. The extent of genetic influences did not differ significantly with respect to sex or between HOC and VEC. The results of bivariate variance decomposition analysis strongly suggest the existence of common genetic factors simultaneously affecting HOC and VEC; 41.8% of the two traits' total residual variance was attributable to the effect of these common genetic factors.     

Role of 67 kDa cell surface laminin binding protein of Leishmania donovani in pathogenesis.

The role that interaction with laminin may play in Leishmania donovani infection was investigated. Binding of (125)I-radiolabeled laminin, in a liquid-phase assay, by the parasite was rapid, saturable, specific, reversible, and of high affinity. Using a Western blotting procedure, a 67 kDa laminin-binding protein (LBP) was identified from the membrane of both the promastigote and amastigote forms of L. donovani. Subsequently, the protein was purified by affinity chromatography. Immunofluorescence with a polyclonal antibody against LBP as well as flow cytometric analysis demonstrated its presence at the parasite surface. After stimulation with phorbol-12-myristate-13-acetate (PMA), U937 cells exhibited the ability to adhere to laminin and LBP specifically inhibited this adhesion. The reduced parasite adhesion after tunicamycin treatment suggested the importance of sugar residues in cell adhesion. Although co-administration of either laminin or LBP or anti LBP antibody reduced parasite virulence, resulting in a lower level of infection in the BALB/c mouse model, an in vitro macrophage culture-enhanced level of infection was observed in the case of laminin-coated parasites. The results collectively suggest a role for LBP in the interaction of the parasite with extracellular matrix elements, which may constitute a basis for the homing of the parasite to its physiological address.     

Types and combinations of axial triradii among the 20 Dhangar castes of Maharashtra,India

Bilateral palmar prints of 3000 males belonging to 20 endogamous Dhangar castes of Maharashtra, India, have been analysed for types and combinations of axial triradii after Cummins and Midlo (1943). Altogether 17 types of axial triradii were found among the Dhangars, but only typest, t′, tt″ occur in appreciable frequencies. The magnitude of intercaste differentiation in respect to axial triradii is considerably low; only 13 caste-pairs (6.84%) out of 190 pairs showed significant difference at the 5% level. Compared to several other palmar elements, like palmar true patterns, main line terminations, palmar flexion creases, the axial triradius has a considerably low level of differentiation among these nomadic, seminomadic and settled Dhangar castes. This suggests that the axial triradii are perhaps more stable compared to many other palmar elements. These results need to be confirmed by data from other populations.     

Intracranial Stereotaxic Cannulation for Development of Orthotopic Glioblastoma Allograft in Sprague-Dawley Rats and Histoimmunopathological Characterization of the Brain Tumor

Glioblastoma is the most common brain tumor that causes significant mortality annually. Limitations of the current therapeutic regimens warrant development of new techniques and treatment strategies in orthotopic animal model for better management of this devastating brain cancer. There are only a few experimental orthotopic models of glioblastoma for pre-clinical testing. In the present investigation, we successfully implanted rat C6 cells via intracranial stereotaxic cannulation in adult Sprague-Dawley rats for development and histoimmunopathological characterization of an advanced orthotopic glioblastoma allograft model, which could be useful for investigating the course of glioblastoma development as well as for testing efficacy of new therapeutic agents. The orthotopic glioblastoma allograft was generated by intracerebral injection of rat C6 cells through a guide-cannula system and after 21 post-inoculation days the brain tumor was characterized by histoimmunopathological experiments. Histological staining and immunofluorescent labelings for TERT, VEGF, Bcl-2, survivin, XIAP, and GFAP revealed the distinct characteristics of glioblastoma in C6 allograft, which could be useful as a target for treatment with emerging new therapeutic agents. Our investigation indicated the successful development of intracranial cannulated orthotopic glioblastoma allograft in adult Sprague-Dawley rats, making it as a useful animal model of glioblastoma for pre-clinical evaluation of various therapeutic strategies for the management of glioblastoma. Special issue in honor of Naren Banik.