Genetic characterization of the highlander Tibetan population from Qinghai-Tibet Plateau revealed by X chromosomal STRs.

Tibetans are considered an East Asian ethnic group and primarily live in the high Tibetan plateau, the western Sichuan and Yunnan mountains of central and southern China, and areas throughout the Himalayas and around the Tibetan plateau. These people exhibit rare molecular machinery that allows them to adapt to hypoxic environments in the Qinghai-Tibet Plateau and make them a potential candidate for providing insights related to medical genetic, molecular medicine and human population studies. In the current study, we have genotyped 549 individuals with Investigator Argus X-12 Kit. For 12 X-STRs, a total of 174 unique alleles were found, among them DXS10134 and DXS10135 were the most polymorphic loci. All of the loci were in Hardy-Weinberg Equilibrium (HWE). The numbers of observed haplotypes in Highlander Tibetans males were 161,112, 96 and 108, respectively, whereas haplotype diversities (HD) were 0.9959, 0.9880, 0.9809 and 0.9873, respectively. The combined discrimination power for males (PDm) was 0.999 999 99701 and for females (PDf) was 0.999 999 999 999 9958. This study represents an extensive report on X chromosomal STR markers variation in the Highlander Tibetans population for forensic applications and population genetic studies.

A preliminary molecular survey of Babesia divergens and first evidence of Theileria annulata in cattle from Saudi Arabia.

BACKGROUND AND AIM: Babesia divergens causes human babesiosis in Europe where the parasite utilizes cattle as animal reservoir and Ixodes ricinus as tick vector. Importation of infected animals and passive carriage of infected ticks through migratory birds can lead to tick/pathogen geographic expansion and emergence of diseases in naïve land. Given the information that Saudi Arabia imports cattle from the European countries and that two global bird flyways pass through the country geographic coordinates, we speculate that B. divergens might be introduced into the Kingdom. Therefore, the aim of this preliminary study was to molecularly detect and characterize B. divergens and other piroplasms (including Theileria spp.) in cattle from Taif district, Kingdom of Saudi Arabia. MATERIALS AND METHODS: Blood samples from 20 cattle residing Taif district were collected, and polymerase chain reaction tested using wide and species-specific primers. Amplicons from a positive genus-wide reaction were purified, sequenced, and analyzed. Phylogenetic trees were constructed, and similarity to existing GenBank zoonotic piroplasms was also assessed. RESULTS: All samples were negative for B. divergens, and only one sample proved positive for Theileria annulata in a wide reaction. Phylogeny clustered our strain with T. annulata from Spanish dog and another one detected in a cow from France. BLAST analysis showed genetic distance from zoonotic piroplasms with identity ranged from 88% to 91%. CONCLUSION: Although B. divergens was not detected, we are not able to rule out or affirm the existence of the pathogen in the country. On the other hand, identifying T. annulata strain with a southern European origin strongly supports our speculation that bovine zoonotic Babesia might be introduced into KSA. This study is not only the first molecular survey of B. divergens but also the first report of the molecular identity of T. annulata in Saudi Arabia. A national-wide bovine and tick surveillance are needed to further prove our speculation.

Physiological and molecular study on the anti-obesity effects of pineapple (Ananas comosus) juice in male Wistar rat.

The present study was performed to assess anti-obesity effects of raw pineapple juice in high fat diet (HFD)-induced fatness. Based on food type, rats were divided into normal diet and HFD groups. When animals of HFD group become obese, they were given pineapple juice along with either HFD or normal diet. Blood biochemistry, liver and muscle gene expressions were analyzed. HFD induced significant elevations in body weight, body mass index (BMI), body fat accumulation, liver fat deposition and blood lipids while juice restored these parameters near to their normal values. Juice significantly decreased serum insulin and leptin while adiponectin was increased. Juice administration downregulated the increment of FAS and SERBP-1c mRNA expression in liver and upregulated HSL and GLUT-2 expressions. The muscular lipolytic CPT-1 expression was upregulted by juice treatment. Pineapple juice, therefore, may possibly be used as anti-obesity candidate where it decreased lipogenesis and increased lipolysis.

Protective and curative effects of the sea cucumber Holothuria atra extract against DMBA-induced hepatorenal diseases in rats.

Oxidative stress is a common mechanism contributing to the initiation and progression of hepatic damage. Hence there is a great demand for the development of agents with potent antioxidant effect. The aim of the present study is to evaluate the efficacy of Holothuria atra extract (HaE) as an antioxidant against 7,12-dimethylbenz[a]anthracene- (DMBA-) induced hepatorenal dysfunction. Experimental animals were divided into two main groups: protective and curative. Each group was then divided into five subgroups pre- or posttreated either with distilled water (DMBA subgroups) or with HaE (200 mg/kg body weight) for seven and fourteen days. Single oral administration of DMBA (15 mg/kg body weight) to Wistar rats resulted in a significant increase in the serum liver enzymes and kidney function's parameters. DMBA increased level of liver malondialdehyde (MDA), decreased levels of reduced glutathione (GSH), glutathione-S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT) in the liver tissue, and induced liver histopathological alterations. Pre- or posttreatment with HaE orally for 14 days significantly reversed the hepatorenal alterations induced following DMBA administration. In conclusion, HaE exhibits good hepatoprotective, curative, and antioxidant potential against DMBA-induced hepatorenal dysfunction in rats that might be due to decreased free radical generation.

Preliminary molecular identification of a predatory bug, Orius albidipennis, collected from ornamental plants.

Orius albidipennis (Reuter) (Hemiptera: Anthocoridae) is a generalist predator used for biological control of insects attacking ornamental plants. Molecular identification of this species using internal transcribed spacer 1 (ITS1) of ribosomal DNA was conducted for the first time. The complete sequence of ITS1 and fragments of its flanking 18S and 5.8S rDNA genes are reported herein. The estimated length of ITS1 of O. albidipennis was 305 bp. This spacer was nearly identical to its counterpart of Orius sp-Taif strain in spite of the difference in their length. The phylogentic relationships were determined using the maximum-likelihood method supported with strong bootstrap probabilities clustering of both taxa together. Further molecular markers could be useful to identify the Taif strain and support its sister relationship to the Egyptian O. albidipennis.

Efficient newly designed primers for the amplification and sequencing of bird mitochondrial genomes.

In the present study, 27 mitochondrial genomes of diverse avian supra-orders were collected from the Genbank database and their genes were aligned separately. From the alignments, the conserved sequences were selected to design novel conserved primers for amplification and sequencing of the different mitochondrial genes. The reproducibility of these primers was tested in the amplification and sequencing of diverse avian supra-order mitochondrial genomes and was confirmed. This method helped in designing a new set of primers to accelerate both the amplification and the sequencing of bird mitogenomes. It also aids in building mitogenome markers in studying the genetic framework of endemic birds as a preliminary strategy for conservation management of them.

Mitochondrial DNA Variability within Uromastyx ornata philbyi (Agamidae: Squamata) from Southwestern Saudi Arabia.

Approximately 2.4 kbp of mitochondrial DNA was sequenced from 9 individuals of Uromastyx ornata philbyi originating from Taif, Namas, Al-Baha, and Jazan in southwestern Saudi Arabia. The sequenced regions cover eight tRNA genes (tRNA(Gln), tRNA(Ile), tRNA(Met), tRNA(Trp), tRNA(Ala), tRNA(Asn), tRNA(Cys), and tRNA(Tyr)) and two protein-coding genes (NADH dehydrogenase subunit 2 and cytochrome b). U. ornata philbyi had an insertion of 170 bp length between tRNA(Gln) and tRNA(Ile) genes. The first 128 bp of this insertion was similar to the one identified earlier in U. ornata ornata and can be folded into a stem-and-loop structure, which was less stable in U. ornata philbyi than in U. ornata ornata, or the second tRNA(Gln) gene. The next 42 bp of the insertion was unique in U. ornata philbyi and additionally retained a stable stem-and-loop structure. Most base substitutions found in the sequenced genes were synonymous transitions rather than transversions. Tree analyses supported the sister group relationship between the two U. ornata subspecies and divided U. ornata philbyi into two groups: Taif+Namas group in the east of Sarawat and Al-Baha+Jazan group in the west of Sarawat. These molecular data are in agreement with current classification of U. ornata.

Timing of a mtDNA gene rearrangement and intercontinental dispersal of varanid lizards.

The mitochondrial genomes of the Komodo monitor (Varanus komodoensis) and the Nile monitor (V. niloticus) were previously shown to have an extensive gene rearrangement. Here, we show that this gene arrangement widely occurs in varanid taxa originated from Africa, Asia and Australasia. Based on phylogenetic relationships of the varanids constructed using mitochondrial DNA sequences encoding the NADH dehydrogenase subunit 2 gene and seven flanking tRNA genes, we estimated their divergence times by the Bayesian method without assuming the molecular clock. The results suggested that the mitochondrial DNA gene rearrangement took place once in an ancestral varanid lineage in the Paleocene or earlier. Our results are more consistent with Cenozoic over-water dispersal of Southeast Asian varanids across the Indonesian Archipelago rather than the Cretaceous Gondwanan vicariance for the origin of Australasian varanids.

The mitochondrial genome of the lizard Calotes versicolor and a novel gene inversion in South Asian draconine agamids.

A complete mitochondrial DNA (mtDNA) sequence was determined for the lizard Calotes versicolor (Reptilia; Agamidae). The 16,670-bp genome with notable shorter genes for some protein-coding and tRNA genes had the same gene content as that found in other vertebrates. However, a novel gene arrangement was found in which the proline tRNA (trnP) gene is located in the light strand instead of its typical heavy-strand position, providing the first known example of gene inversion in vertebrate mtDNAs. A segment of mtDNA encompassing the trnP gene and its flanking genes and the control region was amplified and sequenced for various agamid taxa to investigate timing and mechanism of the gene inversion. The inverted trnP gene organization was shared by all South Asian draconine agamids examined but by none of the other Asian and African agamids. Phylogenetic analyses including clock-free Bayesian analyses for divergence time estimation suggested a single occurrence of the gene inversion on a lineage leading to the draconine agamids during the Paleogene period. This gene inversion could not be explained by the tandem duplication/random loss model for mitochondrial gene rearrangements. Our available sequence data did not provide evidence for remolding of the trnP gene by an anticodon switch in a duplicated tRNA gene. Based on results of sequence comparisons and other circumstantial evidence, we hypothesize that inversion of the trnP gene was originally mediated by a homologous DNA recombination and that the de novo gene organization that does not disrupt expression of mitochondrial genes has been maintained in draconine mtDNAs for such a long period of time.

Mitochondrial genome of Pogona vitticepes (Reptilia; Agamidae): control region duplication and the origin of Australasian agamids.

The complete mitochondrial DNA sequence for an Australian agamid Pogona vitticepes was determined. Twenty-two tRNA genes, two rRNA genes, thirteen protein-coding genes, and two control regions were identified in this mitochondrial genome. The second control region was inserted between NADH dehydrogenase subunits 5 and 6 genes. The duplication of the control region was found in all Australasian agamids examined and was not found in other Asian or African taxa. The two control regions had nearly identical sequences within species but they were divergent among species, suggesting their concerted sequence evolution. Phylogenetic analyses including divergence time estimation without assuming the molecular clock suggested that the duplication of the control region occurred on a lineage leading to the Australasian agamids 25-45 million years ago after their divergence from a Southeast Asian Physignathus cocincinus. Our finding thus supports the recent dispersal origin of Australasian agamids in connection with plate tectonic movement of Australia to the proximity of Southeast Asia.