Mutation in LIM2 Is Responsible for Autosomal Recessive Congenital Cataracts

PurposeTo identify the molecular basis of non-syndromic autosomal recessive congenital cataracts (arCC) in a consanguineous family.MethodsAll family members participating in the study received a comprehensive ophthalmic examination to determine their ocular phenotype and contributed a blood sample, from which genomic DNA was extracted. Available medical records and interviews with the family were used to compile the medical history of the family. The symptomatic history of the individuals exhibiting cataracts was confirmed by slit-lamp biomicroscopy. A genome-wide linkage analysis was performed to localize the disease interval. The candidate gene, LIM2 (lens intrinsic membrane protein 2), was sequenced bi-directionally to identify the disease-causing mutation. The physical changes caused by the mutation were analyzed in silico through homology modeling, mutation and bioinformatic algorithms, and evolutionary conservation databases. The physiological importance of LIM2 to ocular development was assessed in vivo by real-time expression analysis of Lim2 in a mouse model.ResultsOphthalmic examination confirmed the diagnosis of nuclear cataracts in the affected members of the family; the inheritance pattern and cataract development in early infancy indicated arCC. Genome-wide linkage analysis localized the critical interval to chromosome 19q with a two-point logarithm of odds (LOD) score of 3.25. Bidirectional sequencing identified a novel missense mutation, c.233G>A (p.G78D) in LIM2. This mutation segregated with the disease phenotype and was absent in 192 ethnically matched control chromosomes. In silico analysis predicted lower hydropathicity and hydrophobicity but higher polarity of the mutant LIM2-encoded protein (MP19) compared to the wild-type. Moreover, these analyses predicted that the mutation would disrupt the secondary structure of a transmembrane domain of MP19. The expression of Lim2, which was detected in the mouse lens as early as embryonic day 15 (E15) increased after birth to a level that was sustained through the postnatal time points.ConclusionA novel missense mutation in LIM2 is responsible for autosomal recessive congenital cataracts.     

Selection of thermotolerant yeast strains for biomass production from sudanese molasses

Summary Yeast isolates were obtained from different stages in the sugar refining process in an attempt to isolate thermotolerant strains which would grow on a molasses urea medium. Several strains which gave biomass yields of 30–41% at 40° were isolated and identified. Four of these strains were shown to be more resistant to a 15 minute incubation at 55° than three mesophilic strains.     

Receptors on phaeochromocytoma cells for two members of the PP-fold family--NPY and PP

Pancreatic polypeptide (PP) and neuropeptide Y (NPY) belong to a family of regulatory peptides which hold a distinct tertiary structure, the PP-fold, even in dilute aqueous solution. High-affinity receptors, specific for both PP and NPY, are described on the rat phaeochromocytoma cell line, PC-12. The binding of [125I-Tyr36]PP to PC-12 cells was inhibited by concentrations of unlabeled PP which correspond to physiological concentrations of the hormone, 10(-11)-10(-9) mol/l. The affinity of the receptor for the neuropeptide, NPY, was 10(2)-times lower than that of the PP receptor. C-terminal fragments of both PP (PP24-36) and NPY (NPY13-36) were between 10(2)- and 10(3)-times less potent in displacing the radiolabeled 36-amino-acid peptides from their respective receptors. It is concluded that PC-12 cells are suited for structure-function studies of the PP-fold peptides and studies on the cellular events following cellular binding of PP-fold peptides.     

Biotransformation of 7, 12-dimethylbenz (a) anthracene(DMBA): On the genesis of DMBA-trans-dihydrodiols and DMBA- 7- and 12-methyl hydroxylated metabolites

Biotransformation of DMBA in various induced and uninduced S-10 fractions in the presence and absence of hepatic mixed function oxidase inhibitors (α-naphthoflavone, metyrapone, SKF-525A and CS₂) is described. Particular attention is paid to the production of dihydrodiols (DHD) and DMBA-7-and 12-methyl hydroxylated metabolites. It is suggested that DMBA-3, 4-DHD, 8,9-DHD and 5, 6-DHD formation and DMBA-7-and 12-methyl hydroxylations proceed predominantly via different enzyme systems.     

The effect of aging and increasing ascorbate concentrations on respiratory chain activity in cultured human fibroblasts

The specific activities of Complexes I‐III, II‐III, and IV of the respiratory chain, and citrate synthase, were determined in mitochondrial sonicates of six control passage 5 fibroblast cultures, cultivated in growth medium containing fetal calf serum as the only source of ascorbate. The enzymes were also assayed in serially subcultured fibroblasts which were characterized as aged at passage 20 and beyond. Results indicated a significant loss of all enzyme activities in aged cells at passage 20, 25, and 30. Further studies involved maintenance of serially subcultured cells in serum free media to which increasing ascorbate concentrations (100, 200, and 300 µmol 1^?1) were added. Results indicated that ascorbate at 100 µmol 1^?1 was not sufficient to restore any of the enzyme activities in aged cells. An ascorbate concentration of 200 µmol 1^?1 however, could totally restore Complex IV and citrate synthase activities, but had no effect on complexes I‐III and II‐III activities which required 300 µmol 1^?1 ascorbate to be partially or totally restored respectively. In conclusion, this study demonstrates an age related drop in mitochondrial respiratory chain activity in cultured human fibroblasts. Enzyme activities could be completely or partially restored in the presence of double or triple normal human plasma ascorbate concentrations. Copyright © 2010 John Wiley & Sons, Ltd.     

PRDM16, LRP1 and TRPM8 genetic polymorphisms are risk factor for Pakistani migraine patients

BackgroundMigraine is a chronic neurovascular condition characterized by recurring attacks of pulsating headaches. Genome-wide association studies (GWAS) identified many potential loci associated with migraine. To check the association of polymorphisms of PRDM16 (rs2651899), LRP1 (rs11172113), and TRPM8 (rs10166942) with migraine, the first time a case-control study was conducted in understudied Pakistani population.MethodsThe study included 127 migraine patients (21 in migraine with aura and 106 with migraine without aura group) and 120 healthy control subjects from different areas of Punjab, Pakistan. Blood samples were collected from all the participants, and DNA was isolated from the lymphocytes by the modified organic method. Sanger’s sequencing was done for PRDM16 (rs2651899), LRP1 (rs11172113), and TRPM8 (rs10166942) in all the samples to check the genotype. Logistic regression analysis was done using SPSS 20.0 to check the association of these SNPs with migraine susceptibility.ResultsWe found statistically significant differences between case and control group for PRDM16 (rs2651899) at genotypic level (p < 0.001), allelic level (p < 0.001; OR 3.088; 95% CI 2.082–4.579) and for dominant model (p < 0.001; OR 5.437; 95% CI 3.112–9.498). The major findings of this study suggested that PRDM16 rs2651899 is strongly associated with migraine in overall and subgroup analysis of genotypes. LRP1 (rs11172113) showed significant association with migraine except in subgroup comparison. A similar trend of association was found for TRPM8 (rs10166942) however, significant association was found only at the allelic level but no significant difference was seen at the genotypic level between case and control. One novel mutation c.67 + 4436_67 + 4438delA was also identified in the current study near LRP1 (rs11172113) polymorphic site.ConclusionIn this first-ever replication report from Pakistan, PRDM16 (rs2651899) was found as a potential genetic marker in migraine susceptibility while LRP1 (rs11172113) and TRPM8 (rs10166942) showed partial association in subgroup analysis.     

Glicentin is present in the pig pancreas

Specimens from porcine pancreas and ileal mucosa were extracted in acid/ethanol, subjected to gel permeation chromatography, ion-exchange chromatography, enzymatic peptide degradation, reverse-phase HPLC, and analysed for glucagon-like and glicentin-like immunoreactivity by region-specific radioimmunoassays. Results obtained with all methods were consistent with the hypothesis that glicentin is present in the pig pancreas in small amounts.     

e Antigen-antibody system as indicator of liver damage in patients with hepatitis-B antigen.

The clinical relevance of the e antigen-antibody system was investigated in 61 people persistently positive for hepatitis-B surface antigen, including 22 healthy carriers. The e antigen was not detectable in any of the healthy carriers, whereas it was found in 15 out of 28 patients with chronic aggressive hepatitis and two out of 11 with chronic persistent hepatitis. Its presence therefore indicates chronic liver disease but its absence does not exclude it. It may prove to be a particularly useful prognostic aid in chronic persistent hepatitis, since one of the two patients in whom it was found later developed aggressive hepatitis. In contrast, e antibody is of little diagnostic help, for, though it was found mostly in healthy carriers (18;82%), it was also detectable in 9 (23%) of the patients with chronic hepatitis. In 13 (76%) of the patients positive for e antigen Dane particles were seen on electron microscopy, but these were also present in 5 (19%) of the patients positive for e antibody. These findings are consistent with other evidence suggesting that e antigen is not a surface component of the Dane particle, but rather an independent soluble protein manufactured by the host in response to infection with the hepatitis-B virus.     

Phytosociological behaviour and mineral allocation in Panicum turgidum Forssk. along the coast of the Arabian Gulf,Kuwait

This study aims at analyzing the phytosociological behaviour, nutrient element allocation and soil‐plant relations of the Panicum turgidum, which inhabits the arid region of Failaka Island and Jal Az‐Zor along the coastal plain of Kuwait. The above and underground parts of P. turgidum, at 21 different sites distributed across two different localities, as well as surrounding soil, were analyzed. TWINSPAN, DCA and CCA multivariate techniques were applied to the plant species and soil data set, with all statistical calculations using SAS software. Multivariate analysis led to the identification of 4 vegetation groups associated with the distribution of P. turgidum. These groups were: Stipagrostis ciliata‐Cakile arabica, Plantago boissieri, Cyperus conglomeratus and Alhagi graecorum. The above and underground parts of P. turgidum in Failaka showed the presence of a wider range of mineral nutrients, whereas in Jal Az‐Zor there was a greater concentration of heavy metals in the analysed plant parts. These results suggested that Failaka, as an isolated island, exhibited better environmental conditions for the growth of P. turgidum with a more efficient uptake, use and recycling of minerals compared with the Jal Az‐Zor in which heavy metals tended to accumulate in the plants and soil (Br, Zn, Fe and Cu). (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)