Marker discovery and associations with ß-carotene content in Indian dairy cattle and buffalo breeds.

Vitamin A is essential for human health, but current intake levels in many developing countries such as India are too low due to malnutrition. According to the World Health Organization, an estimated 250 million preschool children are vitamin A deficient globally. This number excludes pregnant women and nursing mothers, who are particularly vulnerable. Efforts to improve access to vitamin A are key because supplementation can reduce mortality rates in young children in developing countries by around 23%. Three key genes, BCMO1, BCO2, and SCARB1, have been shown to be associated with the amount of ß-carotene (BC) in milk. Whole-genome sequencing reads from the coordinates of these 3 genes in 202 non-Indian cattle (141 Bos taurus, 61 Bos indicus) and 35 non-Indian buffalo (Bubalus bubalis) animals from several breeds were collected from data repositories. The number of SNP detected in the coding regions of these 3 genes ranged from 16 to 26 in the 3 species, with 5 overlapping SNP between B. taurus and B. indicus. All these SNP together with 2 SNP in the upstream part of the gene but already present in dbSNP (https://www.ncbi.nlm.nih.gov/projects/SNP/) were used to build a custom Sequenom array. Blood for DNA and milk samples for BC were obtained from 2,291 Indian cows of 5 different breeds (Gir, Holstein cross, Jersey Cross, Tharparkar, and Sahiwal) and 2,242 Indian buffaloes (Jafarabadi, Murrah, Pandharpuri, and Surti breeds). The DNA was extracted and genotyped with the Sequenom array. For each individual breed and the combined breeds, SNP with an association that had a P-value <0.3 in the first round of linear analysis were included in a second step of regression analyses to determine allele substitution effects to increase the content of BC in milk. Additionally, an F-test for all SNP within gene was performed with the objective of determining if overall the gene had a significant effect on the content of BC in milk. The analyses were repeated using a Bayesian approach to compare and validate the previous frequentist results. Multiple significant SNP were found using both methodologies with allele substitution effects ranging from 6.21 (3.13) to 9.10 (5.43) µg of BC per 100 mL of milk. Total gene effects exceeded the mean BC value for all breeds with both analysis approaches. The custom panel designed for genes related to BC production demonstrated applicability in genotyping of cattle and buffalo in India and may be used for cattle or buffalo from other developing countries. Moreover, the recommendation of selection for significant specific alleles of some gene markers provides a route to effectively increase the BC content in milk in the Indian cattle and buffalo populations.

Impact of maternal hepatitis B virus coinfection on mother-to-child transmission of HIV.

OBJECTIVES: Despite high hepatitis B virus (HBV) endemicity in various resource-limited settings (RLSs), the impact of maternal HIV/HBV coinfection on infant health outcomes has not been defined. We aimed to assess the prevalence of HBV coinfection among HIV-infected pregnant women and its impact on HIV transmission and infant mortality. METHODS: In this study, the seroprevalence of HBV coinfection was determined among HIV-infected pregnant women enrolled in the Six-Week Extended-Dose Nevirapine (SWEN) India trial. The impact of maternal HIV/HBV coinfection on mother-to-child transmission (MTCT) of HIV and infant mortality was assessed using univariate and multivariate logistic regression analysis. RESULTS: Among 689 HIV-infected pregnant Indian women, 32 (4.6%) had HBV coinfection [95% confidence interval (CI) 3.4%, 5.3%]. HBV DNA was detectable in 18 (64%) of 28 HIV/HBV-coinfected women; the median HBV viral load was 155 copies/mL [interquartile range (IQR) < 51-6741 copies/mL]. Maternal HIV/HBV coinfection did not increase HIV transmission risk [adjusted odds ratio (aOR) 1.06; 95% CI 0.30, 3.66; P = 0.93]. Increased odds of all-cause infant mortality was noted (aOR 3.12; 95% CI 0.67, 14.57; P = 0.15), but was not statistically significant. CONCLUSIONS: The prevalence of active maternal HBV coinfection in HIV-infected pregnant women in India was 4.6%. HIV/HBV coinfection was not independently associated with HIV transmission.

Differentiation of Staphylococcus aureus and Staphylococcus epidermidis by PCR for the fibrinogen binding protein gene.

Mastitis is one of the most common and burdensome diseases afflicting dairy animals. Among other causes of mastitis, staphylococci are frequently associated with clinical and subclinical mastitis. Although Staphylococcus aureus is the predominant species involved, Staphylococcus epidermidis and other coagulase-negative staphylococci are increasingly being isolated from cases of bovine mastitis. Although Staph. aureus and Staph. epidermidis can be easily differentiated based on their biochemical properties, such phenotypic identification is time consuming and laborious. This study aimed to rapidly identify Staph. aureus and Staph. epidermidis. Accordingly, a multiplex PCR was developed and we found that a single gene encoding the adhesin fibrinogen binding protein could be used to identify and differentiate the two species. Consequently, a multiplex reaction combining a triplex PCR for Staph. aureus and a duplex PCR for Staph. epidermidis was standardized, first using bacterial cultures and then with pasteurized milk spiked with live organisms or DNA extracted from the organisms. The test could specifically detect Staph. aureus and Staph. epidermidis even in the presence of a dozen other organisms. The limit of detection for detecting Staph. aureus and Staph. epidermidis separately was 10 to 100 cfu/mL for simplex PCR and 10(4)cfu/mL for multiplex PCR. Conversely, the limit was 10(6)cfu/mL by multiplex PCR for simultaneous detection of both the organisms when spiked into culture medium or pasteurized milk. Overnight enrichment enhanced the assay sensitivity 100-fold. The assay had a high diagnostic sensitivity and specificity. The application of the test was verified on 602 field isolates of staphylococci that had been characterized earlier by phenotypic methods. Importantly, 25 coagulase-negative isolates were identified as Staph. aureus by the multiplex PCR. The test could be adapted for use in clinical diagnostic laboratories.

A high voltage programmable ramp generator.

In this paper, a ramp generator with programmable slope is presented. It consists of a high voltage step generator, followed by integrator. The capacitor and inductor in the integrator are designed such that they can be varied by a microcontroller. This circuit generates two bipolar ramps with fastest speed <1 ns and provides continuous speed variation from 6 to 30 ns for a ramp of 500 V. This is being developed as a part of automated streak camera for deflection of electron beam.

Tribocatalytic behaviour of a TiO2 atmospheric plasma spray (APS) coating in the presence of the friction modifier MoDTC: a parametric study.

Recent engine design and emission trends have led to the commercial use of Atmospheric Plasma Spray (APS) coatings for cylinder liner applications like the TiO2 APS coating. It was shown in our previous work that this type of coating showed better friction results compared to steel lubricated with MoDTC. To further investigate this feature, a parametric study was carried out involving the effect of MoDTC concentration, test temperature, Hertzian contact pressure and the change of counterpart materials from steel balls to ceramic balls (Al2O3 and ZrO2). Ball-on-flat tribotests were carried out on a reciprocating (ball-on-flat) tribometer lubricated with base oil containing MoDTC. Results show that for all the test conditions used including the concentration of MoDTC, test temperature and the contact pressure, lower friction and wear is observed for the TiO2 APS coating compared to reference steel. To explain the low friction behavior, tribofilm compositions were investigated and it was observed that MoS2 is always formed in the case of TiO2 APS with no oxysulphide species. For the reference steel, MoO x S y species are mainly detected in the tribofilms. XPS analyses performed on TiO2 APS flats when the counterpart material was changed from steel balls to ceramic balls suggested the formation of MoS2 (Mo in +iv oxidation state) and Mo-C (Mo in +iv or +ii oxidation state) species with a negligible amount of MoO3 (Mo in +vi oxidation state). It was also shown that a significant amount of molybdenum atoms inside the tribofilm, originating from MoDTC (Mo in +v oxidation state) were reduced in the tribological contact. A mechanism for the decomposition of MoDTC on the basis of tribocatalytic behaviour hypothesized in our previous work was proposed and discussed.

Isolation and characterization of some potential impurities in ropinirole hydrochloride.

Three impurities in ropinirole hydrochloride drug substance at levels approximately 0.06-0.15% were detected by reverse-phase high performance liquid chromatography (HPLC). These impurities were isolated from the drug substance. These impurities were analyzed using reverse-phase HPLC. Based on the spectral data (IR, NMR and MS), structures of these impurities were characterized as 4-[2-(propylamino) ethyl]-1,3-dihydro-2H-indol-2-one hydrochloride (impurity-A), 5-[2-(diropylamino) ethyl]-1,4-dihydro-3H-benzoxazin-3-one hydrochloride (impurity-B) and 4-[2-(diropylamino) ethyl]-1H-indol-2,3-dione hydrochloride (impurity-C). Synthesis of these impurities is discussed.

Household food insecurity is associated with low interferon-gamma levels in pregnant Indian women.

SETTING: Over 20% of tuberculosis (TB) cases during pregnancy occur in India. OBJECTIVE: To determine the association between household food insecurity and interferon-gamma (IFN-γ) levels in pregnancy. DESIGN: Pregnant women in India were administered the Household Food Insecurity Access Scale (HFIAS) questionnaire and underwent an IFN-γ release assay. Logistic regression was used to identify factors associated with food insecurity. RESULTS: Of 538 women, 60 (11%) had household food insecurity, 47 (78%) of which were moderate or severe food insecure. After mitogen stimulation, moderate or severe food insecure women had a median IFN-γ concentration of 4.2 IU/ml (IQR 2.2-9.8) vs. 8.4 IU/ml (IQR 3.0-10) in women with no or mild food insecurity (P = 0.03). In multivariate analysis, higher IFN-γ concentrations were associated with human immunodeficiency virus infection (OR 1.3, 95%CI 0.51-2.1, P = 0.001), and inversely associated with moderate or severe food insecurity (OR -1.6, 95%CI -2.9 to -0.27, P = 0.02) and the number of adults in the household (OR -0.08, 95%CI -0.16 to -0.01, P = 0.03). There was no association between food insecurity and IFN-γ response to Mycobacterium tuberculosis antigen. CONCLUSION: Food insecurity in pregnancy is associated with low IFN-γ levels. There was no association between food insecurity and IFN-γ response to M. tuberculosis antigen, but our study was underpowered to detect this outcome.

Mining precise cause and effect rules in large time series data of socio-economic indicators.

Discovery of cause-effect relationships, particularly in large databases of time-series is challenging because of continuous data of different characteristics and complex lagged relationships. In this paper, we have proposed a novel approach, to extract cause-effect relationships in large time series data set of socioeconomic indicators. The method enhances the scope of relationship discovery to cause-effect relationships by identifying multiple causal structures such as binary, transitive, many to one and cyclic. We use temporal association and temporal odds ratio to exclude noncausal association and to ensure the high reliability of discovered causal rules. We assess the method with both synthetic and real-world datasets. Our proposed method will help to build quantitative models to analyze socioeconomic processes by generating a precise cause-effect relationship between different economic indicators. The outcome shows that the proposed method can effectively discover existing causality structure in large time series databases.

Role of Components in the Formation of Self-microemulsifying Drug Delivery Systems.

Pharmaceutical research is focused in designing novel drug delivery systems to improve the bioavailability of poorly water soluble drugs. Self-microemulsifying drug delivery systems, one among the lipid-based dosage forms were proven to be promising in improving the oral bioavailability of such drugs by enhancing solubility, permeability and avoiding first-pass metabolism via enhanced lymphatic transport. Further, they have been successful in avoiding both inter and intra individual variations as well as the dose disproportionality. Aqueous insoluble drugs, in general, show greater solubility in lipid based excipients, and hence they are formulated as lipid based drug delivery systems. The extent of solubility of a hydrophobic drug in lipid excipients i.e. oil, surfactant and co-surfactant (components of self-microemulsifying drug delivery systems) greatly affects the drug loading and in producing stable self-microemulsifying drug delivery systems. The present review highlighted the influence of physicochemical factors and structural features of the hydrophobic drug on its solubility in lipid excipients and an attempt was made to explore the role of each component of self-microemulsifying drug delivery systems in the formation of stable microemulsion upon dilution.

Sequence length and error analysis of Sequenase and automated Taq cycle sequencing methods.

We have examined DNA sequence error as a function of length using both a manual method of performing reactions with Sequenase and an automated Taq cycle sequencing method. DNA fragments from both methods were separated and analyzed on a sequencer. To determine the sequence of a cosmid insert (35.3 kb), 379 sequences were obtained from a manual Sequenase method, and 354 sequences were obtained from a Taq cycle sequencing method as performed on an automated robotic workstation and sequenced on an automated fluorescent sequencer. A highly redundant consensus of these sequences was obtained and aligned with the individual sequences to determine sequence error over the length of each sequence. The results of this study indicate that error is about 1% per position over the first 350 nucleotides, but increases thereafter to about 17% at 500 nucleotides. This pattern of accuracy was nearly equivalent for manual Sequenase methods and automated Taq cycle sequencing methods. The potential of these methods in large-scale DNA sequencing projects is discussed.

Failure of autoresuscitation in weanling mice: significance of cardiac glycogen and heart rate regulation.

"Autoresuscitation" (AR) is the spontaneous recovery from hypoxic apnea by gasping. We examined aspects of heart function in two situations: 1) the maturationally acquired failure of AR that is characteristic of SWR, but not BALB/c, weanling mice and 2) AR failure in BALB/c mice induced by repeated exposures to anoxia. We determined maturational changes in heart and liver glycogen. Unlike liver glycogen levels, heart glycogen levels in SWR mice differed from those in BALB/c mice. They were consistently much lower throughout maturation and reached a nadir during the brief period when SWR weanling mice are vulnerable to AR failure. Also, rate of cardiac glycogen utilization in vulnerable SWR mice was lower than that of same-aged BALB/c mice and was nil during the latter one-half of the gasping stage when heart function is critical for AR success. Therefore, because glycogen utilization reflects cardiac work, heart failure could explain AR failure in SWR weanlings. Additionally, the increase in hypoxic heart rate that occurs with maturation is developmentally delayed in SWR mice, and this may contribute to their AR failure. Cardiac glycogen was not fully depleted in BALB/c mice during repeated anoxic exposures, indicating other reasons for AR failure. We view these findings as a potential model for the age-related peak in incidence of sudden infant death syndrome.

Widespread occurrence of the Plasmodium falciparum chloroquine resistance transporter (Pfcrt) gene haplotype SVMNT in P. falciparum malaria in India.

The Plasmodium falciparum chloroquine resistance transporter (Pfcrt) K76T mutation and haplotype (amino acids 72-76) and the P. falciparum multidrug resistance 1 (Pfmdr1) mutation (N86Y) were analyzed as markers of chloroquine resistance in the DNAs of 73 blood samples from patients with P. falciparum malaria in India. Seventy of the 73 DNAs had the Pfcrt K76T mutation. Of these, 66 had the SVMNT haplotype and four had CVIET, the African/Southeast Asian haplotype. Only 20 of 69 DNAs had the Pfmdr1 N86Y mutation. It is surprising that the Pfcrt haplotype in India is predominantly SVMNT, rather than that seen in Southeast Asia. The widespread prevalence of the Pfcrt K76T mutation is a cause for concern.

Hypertension: a cause of growth impairment.

The effects of high blood pressure on growth are not fully understood and while hypertension may be associated with failure to thrive, hypertension causing failure to thrive in children is poorly documented. We describe four children presenting with failure to thrive due to hypertension consequent to various aetiologies. Control of hypertension with appropriate therapy resulted in improved growth. The exact pathogenesis of failure to thrive in hypertensive children is not known. These cases demonstrate the importance of careful measurement of blood pressure in children with failure to thrive.

Cataract surgery performed before 800 B.C.

The general belief that the technique of cataract extraction in India in the Sushruta period (800 B.C.) was couching is no longer tenable. A study of the original text suggests that the method was more closely allied to the extracapsular extraction of recent times.

Synthesis of C-glycosylarenes by way of internal reactions of benzylated and benzoylated carbohydrate derivatives.

"Internal" C-glycosylarenes [e.g., (2R,3S,3aS,9bR)-3,3a,5,9b-tetrahydro-3-methoxy- and -3,7-dimethoxy-2-methoxymethyl-2H-furo[3,2-c][2]benzopyran] were prepared by intramolecular reactions of 2-O-benzyl derivatives of methyl 3,5-di-O-methyl-D-xylofuranoside (2) and their conversion into authentic C-glycosylated aromatic systems was investigated. The auxiliary benzylic linkage could not be cleaved by hydrogenolysis; isochroman derivatives (e.g., (3S)-3,4-dihydro-3-[(1R,2R)-2-hydroxy-1,3-dimethoxypropyl]-5-metho xy-1H- 2-benzopyran) were obtained under these conditions. However, oxidation of the primary benzylic position with ruthenium tetraoxide gave the corresponding lactone (a dihydroisocoumarin derivative, e.g., (2R,3S,3aS,9bR)-2,3,3a,9b-tetrahydro-3,7-dimethoxy-2-methoxy - methyl-5H-furo[3,2-c][2]benzopyran-5-one) which could be opened by saponification, thereby leading to a stereochemically unique C-glycosylbenzoic acid derivative. The same type of lactone was obtained directly from a derivative of 2 bearing a sufficiently reactive benzoyl group at O-2 (3,5-dimethoxybenzoyl); this process provides a useful approach to a heterocyclic system present in a variety of natural products. In related studies, the 2-O-phenyl substituent was found to be much less reactive than the 2-O-benzyl group in intramolecular Friedel-Crafts reactions of 2-O-substituted glycofuranosides. The first examples of successful internal C-arylation in the pyranoid series were achieved from 2-O-(3-methoxybenzyl)-D- mannopyranosides; the resulting "internal C-glycosides" [( 2R,3S,4S,4aS,10bS)-2,3,4,4a,6,10b- hexahydro-3,4,8-trimethoxy-2-methoxymethylpyrano[3,2-c][2]benzopyr an and 3,4-bis(benzyloxy)-2-benzyloxymethyl-8-methoxy analog] contain a heterocyclic skeleton closely related to that of the natural product bergenin.

Effect of azadirachtin on Mesocyclops leuckarti sensu lato, vector of dracunculiasis.

Extension or otherwise of characteristic insect development inhibitory effects of the neem sesquiterpenoid azadirachtin have been examined on eggs and nauplii of the crustacean cyclops, M. leuckarti sensu lato, a major vector of guineaworm disease at 0.1 ppm and above doses. Azadirachtin reduced survival of early nauplii (N1 to N3) at 0.1 ppm and above doses. No such effect was observed when eggs and late nauplii (N4 to N5) were exposed. Azadirachtin did not affect the moult cycle when late nauplii (N4 to N5) were exposed continuously till adulthood. At all concentrations tested, adult cyclops exhibited normal mating.

Hyalomma anatolicum anatolicum (Koch, 1844) as a possible vector for transmission of Trypanosoma theileri, Laveran, 1902 in cattle.

Amastigotes, sphaeromastigotes, epimastigotes and trypomastigotes of Trypanosoma theileri, Laveran, 1902 have been observed in the nymphs and adults of Hyalomma anatolicum anatolicum (Koch, 1844). There ticks were obtained from engorged larvae and nymphs collected from a cross-bred bull with detectable parasitaemia. Infection with T. theileri was established in two 6-8-month old cross-bred calves by subcutaneous inoculation of Trypanosomatid flagellates collected from ticks and by feeding of the infected ticks on the ears of these calves.

DMSO resolves certain compressions and signal dropouts in fluorescent dye labeled primer-based DNA sequencing reactions.

Automated base calling algorithms are more sensitive to the quality of the DNA sequencing data than are the labor intensive visual methods of base calling. To improve this quality, data from DNA sequencing reactions have been compared in order to determine the effects of the inclusion of dimethyl sulfoxide (DMSO). Inclusion of 10% DMSO into the reaction cocktail resolves at least one type of sequence compression. This compression may be due to the lack of ability in T7 DNA polymerase to read through certain sequences correctly. The poor quality of these data is seen as radioactive bands or fluorescent signal peaks that have an abnormal alignment, either in the wrong order or as single bands/peaks. The inclusion of DMSO also resolves sequences where the peak signal is absent or severely diminished, leading to a "gap" in the chromatogram profile. DMSO is better than deaza-dITP for resolving certain compressions. Addition of DMSO is a cheaper and more efficient method for high-throughput DNA sequencing than repeating reactions with base analogs.

Soleus motor neuron excitability before and after standing.

H-reflexes have been used to assess the effect of various postures on the excitability of the soleus motor neuronal pool. The purpose of this study was to determine if the excitability of the motor neuron pool, measured via H-reflexes in a seated position, change after a standing protocol in able-bodied individuals. We hypothesized that the excitability of the motor neuronal pool is minimally affected by the standing protocol leading to a reproducible H-reflex. Ten healthy individuals (height = 69.05+/-2.27 inches, weight = 161.7+/-22.44 lbs, age = 27.7+/-7.0 years) participated in the study. Soleus H-reflex recruitment curves were established before and after a standing protocol in a seated position. The standing protocol involved 12 minutes of active upright standing interspersed with 10 minutes of relaxed passive standing in a standing frame, similar to a protocol currently used for spinal cord injured subjects. The maximum M-waves and H-reflex amplitudes were not systematically changed before and after standing. There was also a strong agreement between the H-reflexes and M-waves measured before and after standing (ICC = 0. 99 and .96, respectively). We conclude that the H-reflexes measured in this study were reproducible, indicating that standing had no long lasting effect on the motor neuronal pool excitability. The findings support that the method discussed in this report is appropriate to assess the effects of electrically induced standing on motor neuron pool excitability in individuals with spasticity from spinal cord injury.

Patent ductus arteriosus with a left to right shunt in a 64-year-old asymptomatic male.

We report here the unusual case of a 64 year old male, who was incidentally found to have a patent ductus arteriosus (PDA). The patient was asymptomatic. Echocardiography revealed a moderate sized PDA with normal left ventricular function and mild pulmonary hypertension. As the patient was asymptomatic and was not willing for any form of intervention, he was discharged and is currently under close follow-up. To the best of our knowledge this is the oldest reported case of a PDA in Indian literature.