A novel deletion mutation in LIPH gene causes autosomal recessive hypotrichosis (LAH2)

Autosomal recessive hypotrichosis is a rare hereditary disorder characterized by sparse hair on scalp and rest of the body of affected subjects. Recently, three clinically similar autosomal recessive forms of hypotrichosis [localized autosomal recessive hypotrichosis (LAH)1], LAH2 and LAH3 have been mapped on chromosomes 18q12.1, 3q27.3, and 13q14.11-q21.32, respectively. For these three loci, two genes DSG4 for LAH1 and LIPH for LAH2 have been identified. To date, only five mutations in DSG4 and two in LIPH genes have been reported. In this study, we have ascertained two large unrelated consanguineous Pakistani families with autosomal recessive form of hypotrichosis. Affected individuals showed homozygosity to the microsatellite markers tightly linked to LIPH gene on chromosome 3q27. Sequence analysis of the gene in the affected subjects from both the families revealed a novel deletion mutation in exon 5 (c.659-660delTA) causing frameshift and downstream premature termination codon. All the three mutations identified in the LIPH gene, including the one in this study, are deletion mutations.     

Infection level of the Asian tapeworm (<Emphasis Type="Italic">Bothriocephalus acheilognathi</Emphasis>) in the cyprinid fish, <Emphasis Type="Italic">Schizothorax niger</Emphasis>, from Anchar Lake,relative to season,sex, length and condition factor

Various studies have shown that the Asian fish tapeworm, Bothriocephalus acheilognathi causes great economic loss in hatcheries, fish farms as well as in lakes. In order to understand the seasonal variation of infection in a nutrient-enriched lake, parasitological investigation was carried out in the indigenous cyprinid fish, Schizothorax niger Heckel 1838 from September, 2008 to August, 2009. Overall, this study revealed definite seasonality of infection (p < 0.05), with greater infection in summer (prevalence = 39.5%) and lesser in winter (prevalence = 8.1%). The prevalence among different seasons revealed significant differences (p < 0.05). Sex was not an important factor influencing the prevalence of the Asian tapeworm. A strong positive correlation (Pearson’s correlation, r = 0.7; p = 0.02) between total length of S. niger and number of Asian fish tapeworms was observed. Similarly, a strong positive correlation existed between weight of fish and number of tapeworms (Pearson’s correlation, r = 0.7; p = 0.005). Prevalence and mean abundance were positively and significantly correlated with water temperature (r = 0.8, p < 0.01 and r = 0.8, p < 0.01, respectively). Thus seasonal dynamics, total length and weight of the host significantly influenced the tapeworm infection. The above findings will be useful in devising the appropriate control strategies for the Asian tapeworm in wild fish in Kashmir valley as well as in similar climatic zones of other parts of the world. Also, information from this study will be used to assess the spread and extent of B. acheilognathi which is a potential threat to the indigenous fish fauna of Anchar Lake.     

Spatial learning in transgenic mice expressing human presenilin 1 (PS1) transgenes

Dominant mutations in the Presenilin 1 gene are linked to an aggressive, early-onset form of familial Alzheimer's Disease (FAD). Spatial memory of transgenic (Tg) mice expressing either mutant (lines Tg(M146L)1, Tg(M146L)76, Tg(L286V)198) or wild type (line Tg(PS1wt)195) human PS1 transgenes was investigated in the Morris water maze (WM) test at 6 and 9 months of age. The results showed that the mutated Tg mice had increased swim speed when compared to non-Tg littermates or Tg PS1 wild type mice. The swim speed difference did not, however, significantly affect the spatial learning in the WM test and all groups showed comparable search paths during training and similar spatial bias during probe trials. When re-tested at 9 months, all mice showed significantly improved learning acquisition of spatial information. The lack of progressive spatial learning impairment in mice expressing the mutated human PS1 transgene in the WM does not preclude impairments in other cognitive tasks but suggests that full phenotypic expression of mutant PS1 alleles may require co-expression of human versions of other AD-associated genes.     

A cysteine protease activity from Plasmodium falciparum cleaves human erythrocyte ankyrin

The malaria parasite Plasmodium falciparum undergoes distinct morphologic changes during its 48-h life cycle inside human red blood cells. Parasite proteinases appear to play important roles at all stages of the erythrocytic cycle of human malaria. Proteases involved in erythrocyte rupture and invasion are possibly required to breakdown erythrocyte membrane skeleton. To identify such proteases, soluble cytosolic extract of isolated trophozoites/schizonts was incubated with erythrocyte membrane ghosts or spectrin-actin depleted inside-out vesicles, which were then analyzed by SDS-PAGE. In both cases, a new protein band of 155 kDa was detected. The N-terminal peptide sequencing established that the 155 kDa band represents truncated ankyrin. Immunoblot analysis using defined monoclonal antibodies confirmed that ankyrin was cleaved at the C-terminus. While the enzyme preferentially cleaved ankyrin, degradation of protein 4.1 was also observed at high concentrations of the enzyme. The optimal activity of the purified enzyme, using ankyrin as substrate, was observed at pH 7.0–7.5, and the activity was strongly inhibited by standard inhibitors of cysteine proteinases (cystatin, NEM, leupeptin, E-64 and MDL 28 170), but not by inhibitors of aspartic (pepstatin) or serine (PMSF, DFP) proteinases. Furthermore, we demonstrate that protease digestion of ankyrin substantially reduces its interaction with ankyrin-depleted membrane vesicles. Ektacytometric measurements showed a dramatic increase in the rate of fragmentation of ghosts after treatment with the protease. Although the role of ankyrin cleavage in vivo remains to be determined, based on our findings we postulate that the parasite-derived cysteine protease activity cleaves host ankyrin thus weakening the ankyrin-band 3 binding interactions and destabilizing the erythrocyte membrane skeleton, which, in turn, facilitates parasite release. Further characterization of the enzyme may lead to the development of novel antimalarial drugs.     

A novel locus for alopecia with mental retardation syndrome (APMR2) maps to chromosome 3q26.2-q26.31

Congenital alopecia may occur either alone or in association with ectodermal and other abnormalities. On the bases of such associations, several different syndromes featuring congenital alopecia can be distinguished. Alopecia with mental retardation syndrome (APMR) is a rare autosomal recessive disorder, clinically characterized by total or partial hair loss and mental retardation. In the present study, a five-generation Pakistani family with multiple affected individuals with APMR was ascertained. Patients in this family exhibited typical features of APMR syndrome. The disease locus was mapped to chromosome 3q26.2-q26.31 by carrying out a genome scan followed by fine mapping. A maximum two-point logarithm of odds (LOD) score of 2.93 at theta=0.0 was obtained at markers D3S3053 and D3S2309. Multipoint linkage analysis resulted in a maximum LOD score of 4.57 with several markers, which supports the linkage. The disease locus was flanked by markers D3S1564 and D3S2427, which corresponds to 9.6-cM region according to the Rutgers combined linkage-physical map of the human genome (build 35) and contains 5.6 Mb. The linkage interval of the APMR locus identified here does not overlap with the one described previously; therefore, this locus has been designated as APMR2.     

A novel splice-site mutation in the CDH3 gene in hypotrichosis with juvenile macular dystrophy

Background.  Hypotrichosis with juvenile macular dystrophy (HJMD) is a rare autosomal recessive disorder characterized by sparse hair on the scalp and early blindness. Mutations in the CDH3 gene have been reported to underlie HJMD.
Aim.  To identify a gene responsible for HJMD in a large, four-generation Pakistani family.
Methods.  Genotyping of 13 members of the family, including 6 affected and 7 unaffected members, was carried out using polymorphic microsatellite markers closely linked to the CDH3 on chromosome 16q22.1. To screen for mutations in the CDH3 gene, all of its exons and splice junctions were amplified using PCR from genomic DNA and sequenced directly, using an automated DNA sequencer.
Results.  Microsatellite analysis showed linkage of the family to the CDH3 gene on chromosome 16q22.1. Sequence analysis of the CDH3 gene revealed a novel splice-site mutation (IVS10–1 G→T), leading to probable skipping of exon 11 and a shift in the reading frame.
Conclusion.  The mutation identified here represents the first mutation in the CDH3 gene causing HJMD in a Pakistani population.     

Foci of extramedullary haemopoiesis in a cerebellar haemangioblastoma.

A posterior fossa haemangioblastoma is described which contained foci of extramedullary haemopoiesis. Although secondary polycythemia has been reported with a frequency of between 5 and 30% in patients with haemangioblastomas only five cases have been described previously in the English literature where haemopoietic tissue was identified in the tumour.