Mitochondrial DNA diversity and maternal origins of Pakistani donkey / Diversidade de DNA mitocondrial e origens maternas do burro do Paquistão

Domestic donkey plays a key role as a draft animal in rural economy of Pakistan where its population is increasing every year. The complete mtDNA control region of forty randomly sampled donkeys was PCR- amplified and sequenced bi-directionally using specific primers. Distinct mtDNA haplotypes obtained in the current study (KY446001−KY446011) were subjected to haplotype (h) and nucleotide diversity (π) measures using DnaS as well as to phylogenetic, Network, and AMOVA analyses. There were a total 27 polymorphic sites present within 11 unique mtDNA haplotypes from the studied 40 animals from different regions. Neighbor-joining network and median-joining network both illustrated the splitting of all these haplotypes into two well-defined Nubian and Somali lineages, confirming African maternal origin of Pakistani domestic donkey. Diversity parameters h (0.967± 0.037) and π (0.02917± 0.00307) were found to reveal high levels of genetic diversity in Pakistani donkeys. AMOVA demonstrated only 1% of genetic differences between two mtDNA maternal lineages, pointing to lack of population substructure in Pakistani donkeys as is the case with worldwide domestic donkey population. Pakistani donkeys have African maternal origin and high levels of mtDNA diversity. High genetic diversity may be due to non-selective breeding and heteroplasmy. We herein provide the first report on mtDNA diversity of control region in Pakistani domestic donkey.

O burro doméstico possui um papel fundamental como animal de tração na economia rural do Paquistão, onde a população desse animal está aumentando a cada ano. A região de controle de mtDNA completa de 40 burros amostrados aleatoriamente foi ampliada por PCR e sequenciada bidirecionalmente por intermédio de primers específicos. Haplótipos distintos de mtDNA obtidos no estudo atual (KY446001 − KY446011) foram submetidos a medidas de haplótipo (h) e diversidade de nucleotídeos (π) por meio de DnaS, bem como análises filogenéticas, de rede e AMOVA. Havia um total de 27 sítios polimórficos presentes em 11 haplótipos de mtDNA exclusivos dos 40 animais estudados de diferentes regiões. A rede de união de vizinhos e a rede de união mediana ilustram a divisão de todos esses haplótipos em duas linhagens núbias e somalis bem definidas, confirmando a origem materna africana do burro doméstico do Paquistão. Os parâmetros de diversidade h (0,967 ± 0,037) e π (0,02917 ± 0,00307) revelaram altos níveis de diversidade genética em burros paquistaneses. AMOVA demonstrou apenas 1% de diferenças genéticas entre as duas linhagens maternas de mtDNA, apontando a falta de subestrutura populacional em burros paquistaneses, como é o caso da população mundial de burros domésticos. Os burros paquistaneses têm origem materna africana e altos níveis de diversidade de mtDNA. A alta diversidade genética pode ser por causa da reprodução não seletiva e de heteroplasmia. Aqui, fornecemos o primeiro relatório sobre a diversidade do mtDNA da região de controle em burros domésticos do Paquistão

Mitochondrial DNA diversity and maternal origins of Pakistani donkey

Abstract Domestic donkey plays a key role as a draft animal in rural economy of Pakistan where its population is increasing every year. The complete mtDNA control region of forty randomly sampled donkeys was PCR- amplified and sequenced bi-directionally using specific primers. Distinct mtDNA haplotypes obtained in the current study (KY446001KY446011) were subjected to haplotype (h) and nucleotide diversity () measures using DnaS as well as to phylogenetic, Network, and AMOVA analyses. There were a total 27 polymorphic sites present within 11 unique mtDNA haplotypes from the studied 40 animals from different regions. Neighbor-joining network and median-joining network both illustrated the splitting of all these haplotypes into two well-defined Nubian and Somali lineages, confirming African maternal origin of Pakistani domestic donkey. Diversity parameters h (0.967± 0.037) and (0.02917± 0.00307) were found to reveal high levels of genetic diversity in Pakistani donkeys. AMOVA demonstrated only 1% of genetic differences between two mtDNA maternal lineages, pointing to lack of population substructure in Pakistani donkeys as is the case with worldwide domestic donkey population. Pakistani donkeys have African maternal origin and high levels of mtDNA diversity. High genetic diversity may be due to non-selective breeding and heteroplasmy. We herein provide the first report on mtDNA diversity of control region in Pakistani domestic donkey.

Resumo O burro doméstico possui um papel fundamental como animal de tração na economia rural do Paquistão, onde a população desse animal está aumentando a cada ano. A região de controle de mtDNA completa de 40 burros amostrados aleatoriamente foi ampliada por PCR e sequenciada bidirecionalmente por intermédio de primers específicos. Haplótipos distintos de mtDNA obtidos no estudo atual (KY446001 KY446011) foram submetidos a medidas de haplótipo (h) e diversidade de nucleotídeos () por meio de DnaS, bem como análises filogenéticas, de rede e AMOVA. Havia um total de 27 sítios polimórficos presentes em 11 haplótipos de mtDNA exclusivos dos 40 animais estudados de diferentes regiões. A rede de união de vizinhos e a rede de união mediana ilustram a divisão de todos esses haplótipos em duas linhagens núbias e somalis bem definidas, confirmando a origem materna africana do burro doméstico do Paquistão. Os parâmetros de diversidade h (0,967 ± 0,037) e (0,02917 ± 0,00307) revelaram altos níveis de diversidade genética em burros paquistaneses. AMOVA demonstrou apenas 1% de diferenças genéticas entre as duas linhagens maternas de mtDNA, apontando a falta de subestrutura populacional em burros paquistaneses, como é o caso da população mundial de burros domésticos. Os burros paquistaneses têm origem materna africana e altos níveis de diversidade de mtDNA. A alta diversidade genética pode ser por causa da reprodução não seletiva e de heteroplasmia. Aqui, fornecemos o primeiro relatório sobre a diversidade do mtDNA da região de controle em burros domésticos do Paquistão.

prevalence of chlamydia trachomatis infection among Saudi women attending the infertility clinic in central Saudi Arabia

To determine the prevalence of Chlamydia trachomatis [C. trachomatis] infection among Saudi women, its clinical presentation, and its association to infertility. This study was conducted between October 2012 and July 2013 at King Khalid University Hospital and King Abdulaziz University Hospital, Riyadh, Kingdom of Saudi Arabia. Female patients aged between 19 and 46 years old with infertility problems seen at both hospitals were recruited to join the study. A separate group of female patients without infertility problems was also recruited from both hospitals to serve as controls for the study. Endocervical swabs were collected from both groups of patients, and samples were analyzed using real time polymerase chain reaction. There was a statistically significantly greater prevalence of C. trachomatis infection in infertile women [n=8, 8.0%] compared with the fertile group of women [n=1, 1.0%]. The C. trachomatis infection was significantly correlated to infertility. A significant association between infertility and increased prevalence of C. trachomatis infection is shown in this study, thus, we suggest that screening for Chlamydial infection to be part of the routine investigation for infertility

Carbon isotope fractionation studies of bacterial strain Rhodococcus rhodochrous MTCC 291.

Carbon isotope fractionation associated with the aerobic consumption of propane (C3) were determined using Rhodococcus rhodochrous MTCC 291 bacterial strain to estimate the amount of hydrocarbon oxidized using GC, fractionation of δ13C carbon isotopes of propane and CO2 using GC-C-IRMS and growth of bacteria by UV-Visible Spectrophotometer. The initial δ13C isotopic value of propane was -34 ‰ and after incubation the changes of the isotopic values have been determined on 6th, 10th, 14th, and 17th days. The propane isotopic fractionation value was found to be maximum of -38.0 ‰ with an average value of -36.5 ‰ and a standard deviation of -1.22 ‰. The initial δ13C isotopic value of CO2 was -19.601 ‰. The CO2 isotopic fractionation value was found to be maximum of -29.153 ‰ with an average value of -26.859 ‰ and a standard deviation of -28.338 ‰. The consumption of propane gas was estimated using Gas Chromatograph. The initial concentration of propane in control was found to be 53 ppm. On incubation, the consumption of the propane gas was observed to be of 26 ppm. The carbon isotope fractionation presented here may be applied to estimate the extent of C1-C4 oxidation in natural gas samples, and should prove useful in further studying the microbial oxidation of these compounds in the natural environment.

The effectiveness of using different pulp-capping agents on the healing response of the pulp.

Background: Some of the disadvantages of calcium hydroxide Ca(OH) 2 as pulp-capping material are related to the inflammatory response, and its poor sealing ability. Cyanoacrylate glue was proposed in this study as pulp-capping agent because of its sealing ability, and diprogenta, on the other hand, was proposed because of its anti-microbial and anti-inflammatory effects. The aim of this study is to evaluate and compare the histopathological response of the pulp toward cyanoacrylate and diprogenta in direct pulp capping (DPC). Materials and Methods: Cyanoacrylate, diprogenta, and calcium hydroxide were applied on 20 permanent teeth of 6 rabbits divided into four groups, each contains five teeth, as follow: G1: Diprogenta and calcium hydroxide. G2: Cyanoacrylate. G3: Diprogenta and cyanoacrylate. G4: Calcium hydroxide (control group). Results: A remarkable dentin bridge formation was found in all groups. No pulp necrosis was found in any tooth of the four groups. Pathological reactions of the pulp were found in 25%, 40%, 20%, and 100% of groups 1, 2, 3, and 4, respectively. Differences between study and control groups were found statistically significant ( P < 0.05). Conclusion: All materials used in this study were biocompatible and suggested to be studied further on human teeth. These materials when used in combination with Ca(OH) 2 , may give better results

Evaluation of potable groundwater quality in some villages of Adilabad in Andhra Pradesh, India.

Reconnaissance hydrochemical survey was conducted in some villages of Adilabad district, Andhra Pradesh to assess the quality of groundwater, which is mainly used for drinking purpose. The study consists of the determination of physico-chemical properties, trace metals, heavy metals and rare earth elements in water samples. The data showed the variation of the investigated parameters in samples as follows: pH 6.92 to 8.32, EC 192 to 2706 µS cm-1, TDS 129.18 to 1813.02 ppm. The pH of the waters was within the permissible limits whereas EC and TDS were above the permissible limits of World Health Organization (WHO). Total 27 elements (Li, Be, B, Na, Mg, Al, Si, K, Ca, V, Cr, Mn, Fe, Ni, Co, Cu, Zn, As, Se, Rb, Sr, Mo, Ag, Cd, Sb, Ba and Pb) were analyzed using Inductively Coupled Plasma-Mass Spectrometer (ICP-MS). The concentration of elements in water samples ranged between 0.063 to 0.611 mg l-1 for B, 11.273 to 392 mg l-1 for Na, 5.871 to 77.475 mg l-1 for Mg, 0.035 to 1.905 mg l-1 for Al, 0.752 to 227.893 mg l-1 for K, 11.556 to 121.655 mg l-1 for Ca and 0.076 to 0.669 mg l-1 for Fe respectively. The concentrations of Na, Mg, Al, K, Ca, and Fe exceeded the permissible limits of WHO and BIS guidelines for drinking water quality. In the present study, Bhimavaram, Kazipalli, Kannepalli and Chennur areas of the Adilabad are especially prone to geogenic contamination. Overall water quality was found unsatisfactory for drinking purposes.

Characterization of light gaseous hydrocarbons of the surface soils of Krishna-Godavari basin, India.

Several techniques are used for the exploration of hydrocarbons, of which; the geochemical techniques involving the microbiological technique use the principle of detecting the light hydrocarbon seepage activities for indication of sub-surface petroleum accumulations. A survey was carried out to characterize the light gaseous hydrocarbons seeping in oil and gas fields of Krishna-Godavari basin of Andhra Pradesh. A set of 50 sub-soil samples were collected at depths of about 3 m for geochemical analyses and 1m for microbiological analysis. The microbial prospecting studies showed the presence of high bacterial population for methane 2.5 x 102 to 6.0 x 106 cfu g-1, propane 1x102 to 8.0 x 106 cfu g-1 in soil samples. The adsorbed soil gas analysis showed the presence of moderate to low concentrations of methane (26 to 139 ppb), ethane (0 to 17 ppb), propane (0 to 8 ppb), butane (0 to 5 ppb) and pentane (0 to 2 ppb) in the soil samples of the study area. Carbon isotope analysis for methane (´13C1) ranging from -36.6 to -22.7‰ Pee Dee Belemnite (PDB) suggests these gases are of thermogenic origin. Geo-microbial prospecting method coupled with adsorbed soil gas and carbon isotope ratio analysis have thus shown good correlation with existing oil/ gas fields of Krishna-Godavari basin.

Refractory iron-deficiency anemia in a young patient a rare presentation of celiac disease

Refractory iron deficiency anemia can be an atypical presentation of celiac disease. A 24-year-old male was presented with a six year history of iron deficiency anemia, did not respond to oral iron therapy and received 21 pints of blood; upper endoscopy, duodenal biopsy and serology proved the diagnosis of celiac disease. After gluten-free diet the condition improved within two weeks and regained his normal life after 6 months with the normalization of hematological parameter and serum antibodies. The celiac disease should be considered in patient with refractory iron deficiency anemia.

Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis.

PURPOSE: We describe a simple procedure of Immunoperoxidase (IP) technique, using indigenously raised antibody, to screen corneal scrapings for Acanthamoeba cysts and trophozoites. This study sought to determine the utility of this test in the diagnosis of Acanthamoeba keratitis. METHODS: A high titre polyclonal antibody against a local clinical isolate (axenic) of Acanthamoeba species (trophozoite lysate antigen) was raised in rabbits and used for standardization of IP technique for corneal scrapings. Twenty two smears of corneal scrapings, collected from patients showing Acanthamoeba cysts in corneal scrapings stained with calcofluorwhite (pool-1) and patients showing no cysts in similar scrapings (pool-2), were coded and stained by IP technique by a masked technician. All 22 patients had also been tested for bacteria, fungus, and Acanthamoeba in their corneal scrapings by smears and cultures. IP stained smears were examined for organisms including cysts and trophozoites of Acanthamoeba and background staining by two observers masked to the results of other smears and cultures. The validity of the IP test in detection of Acanthamoeba cysts and trophozoites was measured by sensitivity, specificity, positive predictive value and negative predictive value in comparison (McNemar test for paired comparison) with calcofluor white staining and culture. RESULTS: Based on the readings of observer 1 and compared to calcofluor white staining, the IP test had a sensitivity of 100%, a specificity of 94%, positive predictive value of 80% and negative predictive value of 100%. When compared to culture, the values were 83%, 100%, 100% and 94% respectively. Trophozoites missed in calcofluor white stained smears, were detected in 2 out of 6 cases of culture-positive Acanthamoeba keratitis. The Kappa coefficient of interobserver agreement was determined as fair (30.4%). CONCLUSION: The immunoperoxidase technique is a simple and useful test in the diagnosis of Acanthamoeba keratitis. This can supplement the culture results.

Genetic profile of 81 retinoblastoma patients from a referral hospital in southern India.

PURPOSE: To determine chromosomal abnormalities and inheritance pattern in patients with retinoblastoma from a referral hospital in southern India. MATERIALS AND METHODS: Eighty-one retinoblastoma patients from 78 families were included in this study. Peripheral venous blood was taken for chromosomal analysis and pedigree was ascertained for segregation analysis. RESULTS: Male to female ratio was 1.7:1, 55.56% were bilateral retinoblastoma, the mean age of onset was 12.37 months in bilateral and 33.07 months in unilateral cases (p=0.048). Majority (90.12%) had sporadic inheritance and 6.17% had autosomal dominant inheritance. In chromosomal abnormalities, 8.33% had 13q14 deletion, three cases had de novo balanced translocations. CONCLUSION: The age of onset of the disease was much earlier in the bilateral cases compared to unilateral cases. Sporadic inheritance was predominant while only a small percentage of patients had autosomal dominant inheritance. The percentage of patients with 13q14 deletion was higher than reported in the literature and three novel chromosomal translocations were observed. This is one of the largest series of cases reported from India.

Clinico-epidemiological study of hospitalized cases of severe leptospirosis.

In an attempt to understand the clinical spectrum and pathological and biochemical abnormalities and their prognostic importance in leptospirosis, a prospective study was carried out in Port Blair during September 1996 to August 1997. Out of 80 patients suspected to have leptospirosis, 58 were proved to have current leptospiral infection using serological tests and among these, 14 died giving a case fatality rate of 24.1 per cent. The incidence of the disease showed two separate peaks roughly coinciding with the paddy sowing and harvesting season and the majority of the patients had history of exposure to wet and water logged environment prior to the attack of the disease. The disease presented as two separate clinical syndromes--the hepato-renal form and the pulmonary form though some degree of overlap was present. Hepatic and renal complications occurred in 30 patients each with 26 of them having both. These generally occurred late in the course of the disease and the mortality rate was relatively low. In contrast pulmonary complication occurred quite early and the case fatality rate in those patients was very high (6.7% vs 42.9%). The other complications encountered in the current series of cases were refractory hypotension probably due to myocarditis in 40 per cent and neck stiffness and altered sensorium due to central nervous involvement in 12.1 per cent of the patients. The chances of the patients developing complications were considerably low if treated early and very few of them developed any complications after 2 days of hospitalized treatment. The early occurrence of pulmonary complications indicates a pathogenesis totally different from that responsible for the other complications.