Species identification of Malayan Gaur, Kedah-Kelantan and Bali cattle using polymerase chain reaction-restricted fragment length polymorphism.

Mitochondrial DNA (mtDNA) is a useful genetic marker that can be used for species identification. The cytochrome b (Cyt b) gene is a suitable mtDNA candidate gene for use in phylogenetic analyses due to its sequence variability, which makes it appropriate for comparisons at the subspecies, species, and genus levels. This study was conducted to develop a rapid molecular method for species identification of Malayan gaur (Bos gaurus hubbacki), Kedah-Kelantan (KK) (Bos indicus), and Bali (Bos javanicus) cattle in Malaysia. DNA was extracted from blood samples of 8 Malayan gaurs, 30 KK, and 28 Bali cattle. A set of both specific and universal primers for the Cyt b gene were used in PCR amplification. DNA sequences obtained were then analyzed using BioEdit and Restriction Mapper softwares. The PCR products obtained from Cyt b gene amplification were then subjected to restriction enzyme digestion. The amplification, using both specific and universal primers, produced a 154- and a 603-bp fragment, respectively, in all three species. Two restriction enzymes, NlaIV and SspI, were used to obtain specific restriction profiles that allowed direct identification of Malayan gaur, KK, and Bali cattle. Our findings indicate that all three species can be identified separately using a combination of universal primers and the restriction enzyme SspI.

Rescue by 4-phenylbutyrate of several misfolded creatine transporter-1 variants linked to the creatine transporter deficiency syndrome.

Diseases arising from misfolding of SLC6 transporters have been reported over recent years, e.g. folding-deficient mutants of the dopamine transporter and of the glycine transporter-2 cause infantile/juvenile Parkinsonism dystonia and hyperekplexia, respectively. Mutations in the coding sequence of the human creatine transporter-1 (hCRT-1/SLC6A8) gene result in a creatine transporter deficiency syndrome, which varies in its clinical manifestation from epilepsy, mental retardation, autism, development delay and motor dysfunction to gastrointestinal symptoms. Some of the mutations in hCRT-1 occur at residues, which are highly conserved across the SLC6 family. Here, we examined 16 clinically relevant hCRT-1 variants to verify the conjecture that they were misfolded and that this folding defect was amenable to correction. Confocal microscopy imaging revealed that the heterologously expressed YFP-tagged mutant CRTs were trapped in the endoplasmic reticulum (ER), co-localised with the ER-resident chaperone calnexin. In contrast, the wild type hCRT-1 reached the plasma membrane. Preincubation of transiently transfected HEK293 cells with the chemical chaperone 4-phenylbutyrate (4-PBA) restored ER export and surface expression of as well as substrate uptake by several folding-deficient CRT-1 mutants. A representative mutant (hCRT-1-P544L) was expressed in rat primary hippocampal neurons to verify pharmacochaperoning in a target cell: 4-PBA promoted the delivery of hCRT-1-P544L to the neurite extensions. These observations show that several folding-deficient hCRT-1 mutants can be rescued. This proof-of-principle justifies the search for additional pharmacochaperones to restore folding of 4PBA-unresponsive hCRT-1 mutants. Finally, 4-PBA is an approved drug in paediatric use: this provides a rationale for translating the current insights into clinical trials. This article is part of the issue entitled 'Special Issue on Neurotransmitter Transporters'.

Thickness and optical constants calculation for chalcogenide-alkali metal Se80Te8(NaCl)12 thin film.

Chalcogenide-alkali metal semiconducting thin films of four different thicknesses of Se80Te8(NaCl)12 are deposited from bulk by thermal evaporation technique. The crystallinity of the film improves with increasing of thickness as indicated by the recorded X-ray diffraction patterns. The transmission and reflection spectra are measured in the wavelength range of the incident photons from 250 to 2500nm. The thickness and optical constants of the films are calculated based on Swanepeol method using the interference patterns appeared in the transmission spectra. It is found that the films have absorption mechanism which is an indirect allowed transition. The effect of the film thickness on the refractive index and the high-frequency dielectric constant are studied. With increasing the film thickness, both the absorption coefficient and high-frequency dielectric constant increase while the single-oscillator energy, optical band gap and extinction coefficient decrease.

Thermal annealing effect on the optical properties of Ag10As30S60 thin film.

Chalcogenide Ag10As30S60 thin films are prepared using the thermal evaporation technique from the bulk alloy. Deferential Scanning Calorimetry (DSC) curve reveals two crystallization stages for the bulk. The X-ray examination of the as-prepared and annealed films shows that the sample is crystallized in preferential orientations indicated with peaks corresponding the S8 and ternary AsAg3S3 phases. Transmission spectra show that the as-prepared and annealed films have highly transparent over the visible region. The presence of a sharp absorption edge for all films in the transmission spectra recommends Ag10As30S60 thin films as a good optical filter material. The improvement in transparency upon annealing is due to the enhancement in the crystallinity. The decrease in both optical band gap and refractive index of annealed films after crystallization temperatures is discussed in accordance with the structure changes upon annealing.

Congenital hearing impairment associated with rubella: lessons from Bangladesh.

Infection with rubella virus during pregnancy may cause fetal death or the multiple congenital fetal abnormalities that are known as congenital rubella syndrome (CRS). Studies have demonstrated that congenital hearing impairment is the most frequent abnormality associated with intrauterine rubella infection. In the present study, the first of its kind in Bangladesh, we investigated the presence of rubella antibody in hearing-impaired children in order to understand the possible role of rubella infection in the development of hearing impairment. A total of 198 hearing-impaired children and 200 children without hearing problems were studied. After taking a detailed history from the parents, blood samples were collected from both mothers and children; sera were subjected to enzyme-linked immunosorbent assay (ELISA) for anti-rubella IgG. Rubella antibody was detected in 74% of the hearing-impaired children and in 18% of those with normal hearing: this finding correlated with the presence of rubella antibody in the mothers (67%) of rubella seropositive hearing-impaired children. In contrast, we observed rubella antibody in only 14% of the mothers of the children without hearing problems. Consistent with the presence of antibody, 41% of the seropositive mothers who had hearing-impaired children gave a history of fever and rash during early pregnancy. Our study indicates a strong association between rubella infection and hearing impairment in Bangladeshi children. In addition, it also indicates that infection by rubella virus is common in Bangladesh: this suggests that priority should be given to implementing appropriate measures for the control of rubella.

Collection, analysis and cryopreservation of semen from Malayan gaur (Bos gaurus hubbacki): A preliminary study.

The Malayan gaur (Bos gaurus hubbacki) or Seladang is classified as vulnerable by the International Union for Conservation of Nature and Natural Resources (IUCN). The Malayan gaur is mainly distributed in the tropical woodlands of Peninsular Malaysia and Southern Thailand. The aim of this study was to collect, analyze and cryopreserve the semen of wild Malayan gaur. Transrectal massage (TM) and electroejaculation (EEJ) technique was applied in semen collection of the Malayan gaur. The semen was then cryopreserved in liquid nitrogen using slow freezing technique. Makler counting chamber was used to evaluate sperm concentration and motility, while the sperm viability and morphology of fresh and post-thaw sperm was determined using eosin-nigrosin staining protocol. As a result, we have successfully collected the Malayan gaur semen using EEJ technique. Sperm motility, viability and morphological changes of the post-thaw semen of Malayan gaur were found undesirable due to the complication of the cryopreservation process. On the basis of current study it can be concluded that Malayan gaur bulls semen can be obtain by EEJ with no evidence of rectal trauma. Optimization of the process of cryopreservation for Malayan gaur sperm is needed to maintain the cryoviability of the good sperm quality. The data generated in this study would be useful in conservation of genetic diversity program for Malayan gaur.