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1.
J Dairy Sci ; 102(11): 10039-10055, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31477308

ABSTRACT

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.


Subject(s)
Buffaloes/genetics , Cattle/genetics , Genetic Markers , Milk/chemistry , beta Carotene/analysis , Alleles , Animals , Female , Genotype , India , Polymorphism, Single Nucleotide , Pregnancy , Species Specificity , beta Carotene/genetics
2.
RSC Adv ; 8(27): 15056-15068, 2018 Apr 18.
Article in English | MEDLINE | ID: mdl-35541360

ABSTRACT

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.

3.
Int J Tuberc Lung Dis ; 21(7): 797-803, 2017 07 01.
Article in English | MEDLINE | ID: mdl-28633705

ABSTRACT

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.


Subject(s)
Food Supply , HIV Infections/epidemiology , Interferon-gamma/blood , Mycobacterium tuberculosis/immunology , Antigens, Bacterial/immunology , Cross-Sectional Studies , Female , Humans , India , Interferon-gamma Release Tests , Logistic Models , Multivariate Analysis , Pregnancy , Pregnancy Complications, Infectious/epidemiology , Pregnancy Complications, Infectious/immunology , Prospective Studies , Surveys and Questionnaires
4.
Springerplus ; 5(1): 1625, 2016.
Article in English | MEDLINE | ID: mdl-27722044

ABSTRACT

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.

5.
Indian J Pharm Sci ; 77(3): 249-57, 2015.
Article in English | MEDLINE | ID: mdl-26180269

ABSTRACT

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.

6.
HIV Med ; 15(6): 347-54, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24422893

ABSTRACT

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.


Subject(s)
HIV Infections/transmission , Hepatitis B/epidemiology , Infectious Disease Transmission, Vertical/statistics & numerical data , Adult , Coinfection , Female , HIV Infections/mortality , HIV Infections/virology , HIV-1 , Hepatitis B/virology , Humans , India/epidemiology , Infant , Infant Mortality , Logistic Models , Mothers , Odds Ratio , Pregnancy , Prevalence , Seroepidemiologic Studies , Viral Load , Young Adult
7.
Burns ; 39(6): 1170-7, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23597846

ABSTRACT

Over the past two decades a range of 3D models for human skin have been described. Some include native collagen and intrinsic basement membrane proteins and fibroblasts, others are based on xenogeneic collagen or synthetic supports often without fibroblasts. The aim of this study was to look at the influence of media calcium, basement membrane and fibroblasts on the quality of 3D tissue engineered skin produced using human de-epidermized acellular dermis. In this study we deliberately used Euro skin as the source of the donor dermis to examine to what extent this could provide an effective dermal substrate for producing 3D skin for clinical use. Keratinocytes were cultured in the presence and absence of fibroblasts and both with and without basement membrane on decellularized dermis at calcium concentrations ranging from 250µM to 1.6mM over a period of 14 days. Results showed the formation of a well attached epithelium with many of the features of normal skin in the presence of a basement membrane. This was largely independent of the presence of fibroblasts and not greatly influenced by the concentration of calcium in the media. However there was a clear requirement for physiological levels of calcium in the formation of a stratified epithelium in the absence of a basement membrane.


Subject(s)
Cell Culture Techniques/methods , Skin Transplantation/methods , Skin, Artificial , Tissue Engineering/methods , Analysis of Variance , Basement Membrane/cytology , Calcium/administration & dosage , Calcium/physiology , Collagen/metabolism , Fibroblasts/cytology , Humans , Immunohistochemistry , Keratinocytes/cytology
8.
J Dairy Sci ; 96(5): 2857-65, 2013 May.
Article in English | MEDLINE | ID: mdl-23477820

ABSTRACT

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.


Subject(s)
Bacterial Proteins/genetics , Carrier Proteins/genetics , Genes, Bacterial/genetics , Multiplex Polymerase Chain Reaction/methods , Staphylococcus aureus/genetics , Staphylococcus epidermidis/genetics , Animals , Cattle , Female , Mastitis, Bovine/diagnosis , Mastitis, Bovine/microbiology , Milk/microbiology , Sensitivity and Specificity , Staphylococcal Infections/microbiology , Staphylococcal Infections/veterinary
9.
J Perinatol ; 33(4): 307-12, 2013 Apr.
Article in English | MEDLINE | ID: mdl-22955288

ABSTRACT

OBJECTIVE: To characterize the relationship between peripherally inserted central catheters (PICC) tip positions and associated complications in neonates. STUDY DESIGN: Catheter tip position for 319 infants was classified into superior vena cava (SVC, n=131), inferior vena cava (IVC, n=72), brachiocephalic (BC, n=59), midclavicular (MC, n=49) or iliac. Duration of catheter stay and complication profile was compared between central (SVC/IVC) vs non-central PICC, and between SVC vs IVC, SVC vs BC and SVC vs MC. Kaplan-Meier survival analysis and regression models were used. RESULT: Overall length of catheter stay was similar between central and non-central group. Non-central catheters (n=116) had higher complication rates (47 vs 29%; P=0.001), non-elective removals (45 vs 27%; P=0.002) and shorter time to complication (6.2 vs 11.4 days; P=0.043). This difference was primarily due to the complications encountered in MC group, which had the highest rate of infiltration (P<0.001) and mechanical complications while outcomes were similar among other subgroups. Interestingly, catheter survival probability was similar in all groups for first 4 days. Rate and types of blood stream infections were not related to catheter tip position. CONCLUSION: Non-central PICCs are associated with higher rates of infiltration and mechanical complications when the tip is in MC region. BC catheters may have comparable outcomes to SVC in neonates. A careful risk-benefit analysis is warranted when MC catheters are used in neonates.


Subject(s)
Catheter Obstruction , Catheter-Related Infections , Catheterization, Central Venous , Catheterization, Peripheral , Catheters, Indwelling/adverse effects , Canada , Catheter Obstruction/etiology , Catheter Obstruction/statistics & numerical data , Catheter-Related Infections/epidemiology , Catheter-Related Infections/etiology , Catheterization, Central Venous/adverse effects , Catheterization, Central Venous/methods , Catheterization, Central Venous/statistics & numerical data , Catheterization, Peripheral/adverse effects , Catheterization, Peripheral/methods , Catheterization, Peripheral/statistics & numerical data , Equipment Failure Analysis , Female , Humans , Infant, Newborn , Infant, Premature , Intensive Care, Neonatal/methods , Kaplan-Meier Estimate , Male , Outcome Assessment, Health Care , Retrospective Studies , Risk Assessment
10.
J Environ Sci Eng ; 55(2): 245-52, 2013 Apr.
Article in English | MEDLINE | ID: mdl-25464701

ABSTRACT

The petroleum products have wide range of volatility and are required to be stored in bulk. The evaporation losses are significant and it is a economic as well as environmental concern, since evaporative losses of petroleum products cause increased VOC in ambient air. Control of these losses poses a major problem for the storage tank designers. Ever rising cost of petroleum products further adds to the gravity of the problem. Condensation is one of the technologies for reducing volatile organic compounds emissions. Condensation is effected by condenser, which is basically a heat exchanger and the heat exchanger configuration plays an important role. The horizontal spiral coil heat exchanger is a promising configuration that finds an application in VOC control. This paper attempts to understand underlying causes of emissions and analyse the option of horizontal spiral coil heat exchanger as vent condenser.


Subject(s)
Air Pollution/prevention & control , Petroleum Pollution/prevention & control , Volatile Organic Compounds/chemistry , Equipment and Supplies , Hot Temperature
11.
Indian J Med Microbiol ; 29(4): 406-10, 2011.
Article in English | MEDLINE | ID: mdl-22120803

ABSTRACT

BACKGROUND: An early initiation of antifungal therapy in invasive fungal infections (IFIs) is critical in reducing the high mortality rate. Current diagnosis of fungal infection relies on microscopy, culture, antigen, antibody specific tests and histological diagnosis. However, these tests either lack sensitivity or specificity. There is thus the need for a rapid, specific and accurate diagnostic method. OBJECTIVE: The aim of our study was to establish PCR for the rapid detection of Candida and Aspergillus species in clinical specimens with improved sensitivity and specificity. MATERIALS AND METHODS: A total of 71 proven cases of IFI (confirmed by culture) were collected. A total of 15 healthy, 15 patients suffering from bacterial sepsis and 15 patients with HIV, HBV viral infections were included as controls. Clinical specimens were subjected to a standardized nested amplification to produce Round I (504 bp) and Round II (150 bp) amplicons. Restriction digestion was performed on these products for further identification. RESULTS: Analytical sensitivity was determined using 106-10 CFU/ml of cell suspension. The lower detection limit of the assay was 10 CFU/ml of blood. This test was 100% sensitive and specific with a positive predictive value of 100% and a negative predictive value of 96.7%. CONCLUSION: The assay was found to be effective for the rapid detection of Candida and Aspergillus in clinical specimens.


Subject(s)
Aspergillosis/diagnosis , Candidiasis/diagnosis , Fungemia/diagnosis , Molecular Diagnostic Techniques/standards , Mycology/standards , Polymerase Chain Reaction/standards , Aspergillus/genetics , Aspergillus/isolation & purification , Candida/genetics , Candida/isolation & purification , Early Diagnosis , Humans , Molecular Diagnostic Techniques/methods , Mycology/methods , Polymerase Chain Reaction/methods , Sensitivity and Specificity
12.
Indian J Med Microbiol ; 28(4): 366-9, 2010.
Article in English | MEDLINE | ID: mdl-20966571

ABSTRACT

The present study was designed to evaluate the sensitivity and specificity of liquid culture medium (BioFM broth) for the diagnosis of tuberculous meningitis (TBM) in cerebrospinal fluid (CSF). CSF samples from 200 patients (TBM group = 150 and non-TBM group = 50) were tested for culture of Mycobacterium tuberculosis in BioFM liquid culture medium. Out of 150 TBM cases, 120 were found to be culture positive, indicating a sensitivity of 80% in BioFM broth within 2-3 weeks of inoculation. Positive cultures were also observed for CSF from 32 (64%) out of 50 non-TBM patients in BioFM liquid culture medium within 4 days of sample inoculation. Therefore, according to our study, BioFM broth system yielded 80% sensitivity [95% confidence interval (CI): 67-93%] and 36% specificity (95% CI: 57-98%) for TBM diagnosis. Our results indicate that although BioFM broth allows the detection of positive cultures within a shorter time, it has a high potential for contamination or for the coexistence of M. tuberculosis and non-tuberculous meningitis (NTM). This coexistence may go undetected or potentially lead to erroneous reporting of results.


Subject(s)
Cerebrospinal Fluid/microbiology , Culture Media , Mycobacterium tuberculosis/isolation & purification , Tuberculosis, Meningeal/diagnosis , Adolescent , Adult , Aged , Bacteriological Techniques , Female , Humans , Male , Middle Aged , Mycobacterium tuberculosis/classification , Sensitivity and Specificity , Tuberculosis, Meningeal/microbiology , Young Adult
13.
Rev Sci Instrum ; 81(8): 085106, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20815627

ABSTRACT

A microcontroller based system has been developed for automation of the S-20 optical streak camera, which is used as a diagnostic tool to measure ultrafast light phenomenon. An 8 bit MCS family microcontroller is employed to generate all control signals for the streak camera. All biasing voltages required for various electrodes of the tubes are generated using dc-to-dc converters. A high voltage ramp signal is generated through a step generator unit followed by an integrator circuit and is applied to the camera's deflecting plates. The slope of the ramp can be changed by varying values of the capacitor and inductor. A programmable digital delay generator has been developed for synchronization of ramp signal with the optical signal. An independent hardwired interlock circuit has been developed for machine safety. A LABVIEW based graphical user interface has been developed which enables the user to program the settings of the camera and capture the image. The image is displayed with intensity profiles along horizontal and vertical axes. The streak camera was calibrated using nanosecond and femtosecond lasers.

14.
Int J Tuberc Lung Dis ; 14(8): 1032-8, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20626949

ABSTRACT

SETTING: Polymerase chain reaction (PCR) offers great promise for the rapid, sensitive and specific diagnosis of tuberculous meningitis (TBM). However, the isolation of DNA of high quantity and quality from cerebrospinal fluid (CSF) samples is critical for successful PCR assays. OBJECTIVE: To develop and use a single-tube method for the isolation of PCR-compatible DNA from Mycobacterium tuberculosis using Chelex-100 chelating resin, which does not require organic solvents or detergents. DESIGN: The study focused on the standardisation of a suitable Chelex protocol and its evaluation in 32 CSF samples from TBM and non-TBM subjects. A simultaneous comparison was made with the conventional phenol/chloroform extraction method. RESULT: PCR was found to be more sensitive, more rapid and less technically demanding with the Chelex protocol than the conventional phenol/chloroform extraction method (sensitivity 84.2% vs. 73.6%). CONCLUSION: The single-tube method and the simplicity of the procedure permits early and reliable diagnosis of TBM and makes it an attractive method for routine laboratory assays.


Subject(s)
Cerebrospinal Fluid/microbiology , DNA, Bacterial/analysis , Mycobacterium tuberculosis/genetics , Polymerase Chain Reaction/methods , Resins, Synthetic , Tuberculosis, Meningeal/diagnosis , Adolescent , Adult , Chelating Agents , Child , Diagnosis, Differential , Female , Humans , Male , Middle Aged , Mycobacterium tuberculosis/isolation & purification , Polymerase Chain Reaction/standards , Reproducibility of Results , Sensitivity and Specificity , Tuberculosis, Meningeal/cerebrospinal fluid , Tuberculosis, Meningeal/microbiology , Young Adult
15.
Rev Sci Instrum ; 79(5): 054701, 2008 May.
Article in English | MEDLINE | ID: mdl-18513082

ABSTRACT

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.

16.
Indian J Pharm Sci ; 70(2): 233-6, 2008.
Article in English | MEDLINE | ID: mdl-20046720

ABSTRACT

Duloxetine hydrochloride is a potent dual reuptake inhibitor of serotonin and norepinephrine used to treat major depressive disorders. The present work describes a simple, precise and accurate HPTLC method for its estimation as bulk and in tablet dosage form. The chromatographic separation was carried out on precoated silica gel 60 F254 aluminium plates using mixture of chloroform:methanol (8:1 v/v) as mobile phase and densitometric evaluation of spots was carried out at 235 nm using Camag TLC Scanner-3 with win CAT 1.3.4 version software. The experimental parameters like band size of the spot applied, chamber saturation time, solvent front migration, slit width etc. were critically studied and optimum conditions were evolved. The drug was satisfactorily resolved with Rf value 0.11+/-0.01. The accuracy and reliability of the proposed method was ascertained by evaluating various validation parameters like linearity (40-200 ng/spot), precision (intra-day RSD 0.46-0.75%, inter-day RSD 0.46-1.59%), accuracy (98.72+/-0.20) and specificity according to ICH guidelines. The proposed method can analyse ten or more formulation units simultaneously on a single plate and provides a faster and cost-effective quality control tool for routine analysis of duloxetine hydrochloride as bulk drug and in tablet formulation.

17.
Indian J Pharm Sci ; 70(3): 357-61, 2008.
Article in English | MEDLINE | ID: mdl-20046745

ABSTRACT

A sensitive, simple, specific, precise, accurate and rugged method for determination of enantiomeric purity of S-(-)-1-cyclopropyl-6-fluoro-1,4-dihydro-8-methoxy-7-{4-amino-3,3-dimethylpiperidin-1-yl}-4-oxo-quinoline-3-carboxylic acid hydrochloride monohydrate, WCK 1152, a new drug substance has been developed. The method is based on prederivatization of analyte to diastereomer followed by RP-HPLC using endcapped C-18 stationary phase. Column was maintained at 30 degrees C. The UV/Vis detector was operated at 290 nm. Flow rate of the mobile phase was 1.25 ml/min. The method offers excellent separation of two enantiomers with resolution more than 4 and tailing factor less than 1.5. The method was validated for the quantification of R-(+)-enantiomer impurity, WCK 1153 in the bulk drug. Calibration curves showed excellent linearity over the concentration range of 0.1 to 1.5 mg/ml for WCK 1152 and 0.01 to 0.15 mg/ml for WCK 1153. Precision of the method was 1.13%. Limit of detection and limit of quantitation of the method for WCK 1152 were 0.0006 mg/ml and 0.0018 mg/ml and for WCK 1153 were 0.0007 mg/ml and 0.0021 mg/ml, respectively. Average recovery of the WCK 1153 in WCK 1152 was 94.4%. This method was employed in determining enantiomeric purity of clinical trial batches of WCK 1152.

18.
J Pharmacol Exp Ther ; 321(3): 1161-9, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17371806

ABSTRACT

The effects of nicotine on the tail-flick and hot-plate tests were determined to identify nicotinic receptor subtypes responsible for spinally and supraspinally mediated nicotine analgesia in knockin mice expressing hypersensitive alpha(4) nicotinic receptors (L9'S), in seven inbred mouse strains (C57BL/6, DBA/2, A/2, CBA/2, BALB/cByJ, C3H/HeJ, and 129/SvEv), and in two F1 hybrids (B6CBAF1 and B6D2F1). L9'S heterozygotes were approximately 6-fold more sensitive to the antinociceptive effects of nicotine than the wild-type controls in the hot-plate test but not in the tail-flick assay. Large differences in the effects of nicotine were also observed with both tests for the seven mouse strains. A/J and 129 mice were 6- to 8-fold more sensitive than CBA and BALB mice. In addition, B6CBAF1 hybrid mice were even less sensitive than CBA mice. Nicotinic binding sites were measured in three spinal cord regions and the hindbrain of the inbred strains. Significant differences in cytisine-sensitive, high affinity [(125)I]epibatidine binding site levels (alpha(4)beta(2)(*) subtypes), but not in (125)I-alpha-bungarotoxin binding (alpha(7)(*) subtypes), were observed. Significant negative correlations between cytisine-sensitive [(125)I]epibatidine binding and nicotine ED(50) for both tests were noted. Our results indicate that alpha(4)beta(2)(*) acetylcholine nicotinic receptors (nAChR) are important in mediating nicotine analgesia in supraspinal responses, while also showing that alpha(4)beta(2)(*)-nAChR and at least one other nAChR subtype appear to modulate spinal actions.


Subject(s)
Analgesics/pharmacology , Pain/physiopathology , Receptors, Nicotinic/physiology , Alkaloids/metabolism , Analgesics/metabolism , Animals , Azocines/metabolism , Binding, Competitive/drug effects , Bridged Bicyclo Compounds, Heterocyclic/metabolism , Bungarotoxins/metabolism , Cell Membrane/drug effects , Cell Membrane/metabolism , Male , Mecamylamine/pharmacology , Mice , Mice, Inbred BALB C , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Inbred DBA , Mice, Inbred Strains , Mice, Knockout , Morphine/pharmacology , Nicotine/pharmacology , Nicotinic Antagonists/pharmacology , Pain/metabolism , Pain/prevention & control , Pyridines/metabolism , Quinolizines/metabolism , Reaction Time/drug effects , Receptors, Nicotinic/genetics , Spinal Cord/metabolism
19.
J Pharm Biomed Anal ; 43(4): 1587-93, 2007 Mar 12.
Article in English | MEDLINE | ID: mdl-17207602

ABSTRACT

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.


Subject(s)
Antiparkinson Agents/analysis , Indoles/analysis , Antiparkinson Agents/chemistry , Chromatography, High Pressure Liquid/methods , Drug Contamination , Indoles/chemistry , Magnetic Resonance Spectroscopy , Mass Spectrometry , Molecular Structure , Spectrophotometry, Infrared , Spectroscopy, Fourier Transform Infrared
20.
Electromyogr Clin Neurophysiol ; 44(5): 259-64, 2004.
Article in English | MEDLINE | ID: mdl-15378863

ABSTRACT

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.


Subject(s)
H-Reflex/physiology , Motor Neurons/physiology , Muscle, Skeletal/physiology , Posture/physiology , Adult , Electromyography , Female , Humans , Leg/physiology , Male , Reference Values , Reproducibility of Results , Tibial Nerve/physiology
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