Unprecedented 2D GNR-CoB nanocomposite for detection and degradation of malachite green - A computational prediction of degradation pathway and toxicity.

In the present work, we have synthesized a novel 2D GNR-CoB composite and was applied it for electrochemical sensing and photocatalytic degradation of the malachite green (MG). The physicochemical properties of the 2D GNR-CoB were analyzed using X-ray diffraction, Transmission electron microscopy, Energy dispersive X-ray diffraction which depicts the morphological and crystalline nature of the prepared composite. The pencil graphite electrode modified with 2D GNR-CoB composite showed excellent electrochemical response for MG detection with a LOD of 1.92 nM, linear range of 25-350 nM with a high sensitivity of 1.714 µA µM-1 cm-2. Besides, the 2D GNR-CoB modified PGE exhibited good recovery for the detection of MG in real samples such as green peas and lady's fingers. Furthermore, the 2D GNR-CoB modified electrode showed excellent photocatalytic activity for the degradation of MG. It suggests that under visible light, GNR-CoB material generates superoxide (·O2-) and hydroxyl (·OH) radicals for MG degradation. The prepared composite showed an efficiency of 91.28% towards the degradation of MG. Based on the experimental analysis and density functional theory calculations, a photocatalytic degradation mechanism pathway for MG is proposed. A quantitative structure-activity relationship study was used to examine the toxicity of the degradation intermediates.

Graphene nanoribbon: An emerging and efficient flat molecular platform for advanced biosensing.

Graphene nanoribbons (GNRs) are lengthened one-dimensional monolayer strips of graphene and have a hexagonal honeycomb lattice structure. The captivating properties like electrical conductivity, emerging band gap, optical property, thermal conductivity, high mechanical strength, and ultrahigh surface area make them a better candidate for biomedical applications. The properties can be significantly reformed and controlled by altering the edge functionalities and geometry. The exhibition of a wide potential window coupled with an ultra-high surface area to host sensing element makes GNR an excellent biosensing platform. Consequently, biosensing is one of the most explored applications of GNR. This review presents an overview of the characteristics, methods of synthesis, and biosensing applications of GNR. Overall, GNR is considered a promising platform for efficient signal transduction compared to conventional biosensing platforms. Further, it offers high electrical conductivity, large surface area, high adsorption, synergistic effects with combined materials, fast response, sensitivity, and selectivity.

Gene therapy with plasmids encoding IFN-ß or IFN-α14 confers long-term resistance to HIV-1 in humanized mice.

Because endogenous interferon type I (IFN-I) produced by HIV-1 infection might complicate the analysis of therapeutically administered IFN-I, we tested different humanized mouse models for induction of IFN-I during HIV-1 infection. While HIV-1 induced high levels of IFN-α in BLT mice, IFN-I was undetectable following infection in the Hu-PBL mouse model, in which only T cells expand. We therefore tested the effect of treatment with Pegylated IFN-2 (pegasys), in Hu-PBL mice. Pegasys prevented CD4 T cell depletion and reduced the viral load for 10 days, but the effect waned thereafter. We next expressed IFN-I subsets (IFN-α2, -α6, -α8, -α14, and -ß) in Hu-PBL mice by hydrodynamic injection of plasmids encoding them and 2 days later infected the mice with HIV-1. CD4 T cell depletion was prevented in all subtypes of IFN-I-expressing mice by day 10. However, at day 40 post-infection, protection was seen in IFN-ß- and IFN-α14-expressing mice, but not the others. The viral load followed an inverse pattern and was highest in control mice and lowest in IFN-ß- and IFN-α14-expressing mice until day 40 after infection. These results show that gene therapy with plasmids encoding IFN-ß and -α14, but not the commonly used -α2, confers long-term suppression of HIV-1 replication.

Vacuum epidural cyst with acute neurological presentation. A case report.

The widespread use of MRI in the assessment of low back pain has led to increased detection of degenerative cysts of the spine, which was essentially a surgical diagnosis earlier. The awareness of degenerative cysts, the significance of their role in the etiology of radicular and back pain and their effective management is evolving. We describe a case of bilateral, gas-filled lumbar facet synovial/ ganglion cysts causing focal arachnoid inflammation and lateral lumbar canal stenosis.

Is screening for diabetes among tuberculosis patients feasible at the field level?

SETTING: Seventeen peripheral health institutions (PHI) in Kolar district (population: 0.5 million), South India. OBJECTIVE: To assess the feasibility and results of screening patients with tuberculosis (TB) for diabetes mellitus (DM) at peripheral level. DESIGN: From January to September 2012, all TB patients were assessed for DM. Those with unknown DM status were screened for the disease (free of charge) by trained laboratory technicians at each PHI, using a glucometer supplied by the national programme on a capillary blood sample. Those with fasting blood glucose (FBG) ≥ 126 mg/dl (≥7 mM) were diagnosed as DM-positive. RESULTS: Of 362 TB patients, 358 (99%) were assessed for DM and 62 (17.1%) had the diseases-53 (14.6%) had a previous history of DM and 9 (2.9%) were newly diagnosed. All new DM patients were enrolled into DM care. Higher DM prevalence was found among TB patients aged ≥40 years, smokers and those with smear-positive pulmonary TB. To detect a new case of DM, the number needed to screen (NNS) among TB patients was 40. CONCLUSION: Screening of TB patients for DM was feasible and effective in a peripheral setting. The availability of trained laboratory technicians and free services at every PHI made the intervention feasible. The study has contributed towards a national policy decision in this regard.

Activity of hydrolytic enzymes in various regions of normal human brain tissue.

The activity of six hydrolytic enzymes-carboxyl esterase, acid phosphatase, alkaline phosphatase, ß-galactosidase, ß-glucosidase and ß-hexosaminidase, were studied in different regions of the normal human brain tissue obtained at autopsy. Protein estimation and activities of the hydrolytic enzymes with respective substrates were assayed by spectrophotometric and spectroflourometric methods. Amongst the eight regions of the brain-frontal, parietal, occipital, temporal, thalamus, cerebellum and hippocampus, the pineal gland showed highest activity for all hydrolytic enzymes studied except for carboxyl esterase. Among six hydrolases studied, hexosaminidase exhibited highest activity in all regions of the human brain while alkaline phosphatase activity was the least amongst all regions studied. A majority of the enzymes studied showed higher activity in gray matter as compared to the white matter except acid phosphatase and ß-glucosidase which exhibited higher activity in the white matter. The most significant finding in the present study was the high activity of all hydrolytic enzymes noted in the pineal gland as compared to all other regions of the human brain. Such a finding has not been hitherto reported earlier in human brain tissue samples. If the specific activities of these enzymes are to be considered as any functional index, then pineal gland may be more metabolically active tissue with respect to the hydrolytic function as compared to the other regions of the brain.

Synthesis of N-homobicyclic dideoxynucleoside analogues.

Syntheses of six N-homobicyclic dideoxynucleoside analogues are described. The reaction of mannose diacetonide with trimethylsulfoxonium iodide gave a mixture of diastereomeric hydroxymethyl mannose diacetonides in a ratio of 2:5, which was separated by fractional crystallization. The two stereoisomers were converted to bicyclic furanolactols each of which was coupled with three nucleoside bases. Further debenzylations gave the six target N-homobicyclic dideoxynucleosides.

Purification and characterization of carboxylesterase from the seeds of Jatropha curcas.

Carboxylesterases are hydrolases which catalyze the hydrolysis of various types of esters. Carboxylesterase from the seeds of Jatropha curcas has been purified to homogeneity using ammonium sulfate fractionation, CM-cellulose chromatography, Sephadex G-100 chromatography and preparative polyacrylamide gel electrophoresis (PAGE). The homogeneity of the purified enzyme was confirmed by PAGE, iso-electrofocusing and SDS-PAGE. The molecular weight of the purified enzyme was determined by both gel-permeation chromatography on Sephadex G-150 and SDS-PAGE. The molecular weight determined by Sephadex G-150 chromatography and SDS-PAGE both in the presence and absence of 2-mercaptoethanol was 31 kDa. The isoelectric point of the purified enzyme was found to be 8.9. JCSE-I (J. curcas seed esterase-I) was classified as carboxylesterase on the basis of substrate and inhibitor specificity. The K(m) of JCSE-I with 1-naphthyl acetate, 1-naphthyl propionate, 1-naphthyl butyrate and 2-naphthyl acetate as substrates were found to be 0.0,794, 0.0,658, 0.0,567 and 0.1 mM, respectively. The enzyme exhibited an optimum temperature of 45 °C and an optimum pH of 6.5. The enzyme was stable up to 15 min at 65 °C. The enzyme was resistant towards carbamates (carbaryl and eserine sulfate) and sulphydryl inhibitors (p-chloromercuricbenzoate, PCMB) and inhibited by organophosphates (dichlorvos, parathion and phosphamidon).

Carboxylesterases from the seeds of an underutilized legume, Mucuna pruriens; isolation, purification and characterization.

Two carboxylesterases (ME-III and ME-IV) have been purified to apparent homogeneity from the seeds of Mucuna pruriens employing ammonium sulfate fractionation, cation exchange chromatography on CM-cellulose, gel-permeation chromatography on Sephadex G-100 and preparative PAGE. The homogeneity of the purified preparations was confirmed by polyacrylamide gel electrophoresis (PAGE), gel-electrofocussing and SDS-PAGE. The molecular weights determined by gel-permeation chromatography on Sephadex G-200 were 20.89 kDa (ME-III) and 31.62 kDa (ME-IV). The molecular weights determined by SDS-PAGE both in the presence and absence of 2-mercaptoethanol were 21 kDa (ME-III) and 30.2 kDa (ME-IV) respectively, suggesting a monomeric structure for both the enzymes. The enzymes were found to have Stokes radius of 2.4 nm (ME-III) and 2.7 nm (ME-IV). The isoelectric pH values of the enzymes, ME-III and ME-IV, were 6.8 and 7.4, respectively. ME-III and ME-IV were classified as carboxylesterases employing PAGE in conjunction with substrate and inhibitor specificity. The K(m) of ME-III and ME-IV with 1-naphthyl acetate as substrate was 0.1 and 0.166 mM while with 1-naphthyl propionate as substrate the K(m) was 0.052 and 0.0454 mM, respectively. As the carbon chain length of the acyl group increased, the affinity of the substrate to the enzyme increased indicating hydrophobic nature of the acyl group binding site. The enzymes exhibited an optimum temperature of 45°C (ME-III) and 37°C (ME-IV), an optimum pH of 7.0 (ME-III) and 7.5 (ME-IV) and both the enzymes (ME-III and ME-IV) were stable up to 120 min at 35°C. Both the enzymes were inhibited by organophosphates (dichlorvos and phosphamidon), but resistant towards carbamates (carbaryl and eserine sulfate) and sulphydryl inhibitors (p-chloromercuricbenzoate, PCMB).

Preparation and in vitro characterization of mucoadhesive hydroxypropyl guar microspheres containing amlodipine besylate for nasal administration.

Amlodipine besylate microspheres for intranasal administration were prepared with an aim to avoid first-pass metabolism, to achieve controlled blood level profiles and to improve therapeutic efficacy. Hydroxypropyl Guar, a biodegradable polymer, was used in the preparation of microspheres by employing water in oil emulsification solvent evaporation technique. The formulation variables were drug concentration, emulsifier concentration, temperature, agitation speed and polymer concentration. All the formulations were evaluated for particle size, particle shape and surface morphology by scanning electron microscopy, percentage yield, drug entrapment efficiency, in vitro mucoadhesion test, degree of swelling and in vitro drug diffusion through sheep nasal mucosa. The microspheres obtained were free flowing, spherical and the particles ranged in size from 13.4±2.38 µm to 43.4±1.92 µm very much suitable for nasal delivery. Increasing polymer concentration resulted in increased drug entrapment efficiency and increased particle size. Amlodipine besylate was entrapped into the microspheres with an efficiency of 67.2±1.18 % to 81.8±0.64 %. The prepared microspheres showed good mucoadhesion properties, swellability and sustained the release of the drug over a period of 8 h. The data obtained were analysed by fitment into various kinetic models; it was observed that the drug release was matrix diffusion controlled and the release mechanism was found to be non-Fickian. Stability studies were carried out on selected formulations at 5±3°, 25±2°/60±5% RH and 40±2°/75±5% RH for 90 days. The drug content was observed to be within permissible limits and there were no significant deviations in the in vitro mucoadhesion and in vitro drug diffusion characteristics.

Distribution of malignant neoplasms reported at different pathology centers and hospitals in Jaipur, Rajasthan.

BACKGROUND: Cancer data from Rajasthan are limited. Only two studies, one from Western Rajasthan, and the other from Eastern Rajasthan have been published by Sharma et al. in 1992 and 1996. AIMS: To put the cancer profile from this region in proper perspective, we conducted the present study on the patterns of various malignancies in Jaipur region, i.e., Eastern Rajasthan. SETTING AND DESIGN AND MATERIAL AND METHODS: The study spans over one and half decade (1990-2004) and is based on a retrospective six-year sample analysis of approximately 200,000 histopathological and cytological reports for the years 1990, 1991, 1996, 1999, 2001 and 2004. RESULTS: A total of 21,868 cancers were recorded in the six sample years. There were 59.11% (12,926) males and 40.89% (8942) females, with the male to female ratio being 1.45:1. Organ wise, lung (8.45%), prostate (7.12%), brain (6.04%), urinary bladder (5.31%), esophagus (4.67%) and tongue (4.60%) are most common sites involved in males with regard to frequency, whereas breast (20.44%), cervix (14.99%), ovary (4.35%), brain (3.80%), esophagus (3.67%), uterus (3.01%) and rectum (2.80%) are common sites for malignancies in females. CONCLUSIONS: Significant findings were a higher frequency of cancers of the prostate, urinary bladder, and brain in males along with gall bladder cancers in females. Our figures have been compared with the national data.

Postgraduate trainees' assessment of the educational value of ward rounds in obstetrics and gynaecology.

Ward rounds have been regarded as a traditional forum for the teaching and training of trainees. The aim of this research project was to look at what role ward rounds play in the medical education of trainees in obstetrics and gynaecology. A questionnaire comprising 10 statements devised around the theme of quality of medical education and the overall experience of ward rounds was distributed at the Specialist Registrars monthly training days. A total of 46 completed questionnaires were received. The ward rounds took place at least twice weekly at the Welsh Deanery hospitals, however, none of the trainees regarded these ward rounds as 'teaching' ward rounds. A total of 32 (70%) of the trainees disagreed or were uncertain that they learnt something new on ward rounds each day and 34 (74%) of trainees agreed or strongly agreed that in the presence of a consultant, SpRs are not given a chance to lead a ward round. The educational role of the ward rounds does not seem fully utilised in the Welsh Deanery Hospitals and needs to be developed further to suit the needs of junior doctors.

A remarkable accelerating effect of Ag-salt on intramolecular cyclization of o-(1-alkynyl)benzenesulfonamides.

Herein, we report transition metal-catalyzed intramolecular cyclization of o-(1-alkynyl)benzenesulfonamides to afford 3-substituted benzothiazines regioselectively via a C-N bond forming reaction and Cu-catalyzed sequential C-N and C-C bond formation leading to the corresponding 3,4-disubstituted derivatives.

Study of hydroxy propyl guar derivative for its gelling property and it's use in the formulation of tenoxicam gels.

Gels of tenoxicam 1% w/w were formulated using 2% w/w hydroxy propyl guar derivative and 3% w/w sodium carboxy methyl cellulose as gelling agents. A detailed rheological investigation was carried out to study the influence of preservative, drug and preservative, solvent system and the preservative, drug, solvent system and the preservative on the pseudoplastic behaviour of polymers. Hydroxy propyl guar derivative in 2% w/w strength resulted in gels with a higher pseudoplastic index value of 3.383 in contrast to an index value of 1.797 for a 3% w/w sodium carboxy methyl cellulose gels of a similar composition. The gels were stored at different temperatures and variations in pH values were recorded. Hydroxy propyl guar derivative based gels revealed variations in pH values over a narrow range in contrast to sodium carboxy methyl cellulose gels. The gels were subjected to short term stability studies by storing gels at refrigerated temperature, lab temperature, at 37 degrees C and at 45 degrees C. Gels based on hydroxy propyl guar derivative revealed better drug keeping qualities in contrast to sodium carboxy methyl cellulose stabilized gels. Release studies of tenoxicam from formulations across hairless albino mice skin revealed a zero order drug release pattern from both the formulations.

Differentiation of Boc- alpha,beta- and beta,alpha-peptides and a pair of diastereomeric beta,alpha-dipeptides by positive and negative ion electrospray tandem mass spectrometry (ESI-MS/MS).

Positive and negative ion electrospray ionization (ESI) tandem mass spectral study of a new series of hybrid peptides, viz, BocN-alpha,beta-peptides and BocN-beta,alpha-peptides, synthesized from C-linked carbo-beta3-amino acids [Caa (S)] and L-Ala has been carried out. The alpha,beta-peptides have been differentiated from beta,alpha-peptides by the collision-induced dissociation (CID) of [M + H]+ and [M - H]- ions in positive and negative ion ESI-MS respectively. The fragment ion [M + H - C(CH3)3 + H]+ formed from [M + H]+ ions by the loss of 2-methyl-prop-2-ene in alpha,beta-peptides with L-Ala at the N-terminus is insignificant or totally absent for beta,alpha-peptides which have the Caa (S) at N-terminus. The fragment ion [M - H-C(CH3)3OH - HNCO]- formed from [M - H]- of beta,alpha-peptide acids is totally absent for alpha,beta-peptide acids. This has been attributed to the absence of the beta-methylene group in alpha,beta-peptides, and the participation of the beta-methylene group in the loss of HNCO in beta,alpha-peptide acids is confirmed by the deuteration experiments. The CID of [M + H-Boc + H]+ ions of these peptides also produce characteristic fragmentation. In the CID spectra of alpha,beta-peptides, the b(n)+ ions and the resulting y(n)+ ions occur at a mass difference of 243 and 71 Da corresponding to the successive losses of Caa and L-Ala, whereas a mass difference of 71 and 243 Da is observed for beta,alpha-peptides. In contrast to the CID of protonated peptides, the CID of [M - H]- ions of the alpha,beta- and beta,alpha-peptide acids do not give b(n)- ions and show abundant z(n) (-) ions. Further, a pair of diastereomeric dipeptide esters and acids have been distinguished by the CID of [M + H]+ ions. The loss of 2-methyl-prop-2-ene is more pronounced for Boc-NH-Caa(R)-D-Ala-OCH3 (21) and Boc-NH-Caa(R)-D-Ala-OH (23) with Caa (R) at the N-terminus, whereas it is totally absent for Boc-NH-Caa (S)-D-Ala-OCH3 (22) and Boc-NH-Caa(S)-D-Ala-OH (24) peptides, which have Caa (S) at the N-terminus. Thus, on the basis of our previous and present studies, we propose that the CID of [M + H]+ ions provides a simple and useful method for distinguishing the configuration of Caa (S or R) at the N-terminus of BocN-carbo beta,alpha- and beta,beta-dipeptides.

Lentiviral delivery of short hairpin RNAs protects CD4 T cells from multiple clades and primary isolates of HIV.

Viral heterogeneity is a major hurdle for potential therapeutic use of RNA interference (RNAi) against HIV-1. To determine the extent of RNAi tolerance to mutations, we tested 3 viral target sites with differing propensity for mutations: a highly variable rev sequence, a gag sequence conserved only among clade B isolates, and a vif sequence highly conserved across clades. Lentiviral expression of all 3 shRNAs inhibited replication of the homologous HIV(IIIB) strain. However, they differed in their ability to protect primary CD4 T cells against multiple isolates within and across HIV clades. The least conserved rev sequence inhibited only 2 of 5 clade B isolates. The gag sequence (conserved within clade B) protected 5 of 5 clade B isolates but not other clade viruses with 2 or 3 mutations in the central region. In contrast, the vif sequence, which was conserved across clades except for single mutations at positions 14 and 17, inhibited viruses from 5 different clades. Moreover, siRNAs with introduced mutations at sites of gag sequence polymorphisms showed reduced antiviral activity, whereas mutations in vif siRNA only modestly decreased silencing. Thus, although 1 or 2 mutations at peripheral sites are tolerated, mutations in the central target cleavage region abolish RNAi activity.

Genetic typing of bovine viral diarrhoea virus isolates from India.

Thirteen BVDV isolates collected in four geographic regions of India between 2000 and 2002 were typed in 5'-UTR. To confirm results of genetic typing, selected viruses were also analysed in the N(pro) region. Phylogenetic analysis revealed that all Indian BVDV isolates belong to BVDV-1b (Osloss-like group). Despite a long distance between the farms from which the viruses were isolated there was no correlation between the origin of viral isolates and their position in a phylogenetic tree. Higher genetic similarity of Indian BVDV isolates was observed most probably due to the uncontrolled movement of cattle as well as the uncontrolled use of semen from bulls for breeding of local and farm cattle in different states of India.

Affinity labeling of rat cytochrome P450C24 (CYP24) and identification of Ser57 as an active site residue.

25-hydroxyvitamin D(3)- or 1alpha,25-dihydroxyvitamin D(3)-24R-hydroxylase (cytochromeP450C24 or CYP24) has a dual role of removing 25-OH-D(3) from circulation and excess 1,25(OH)(2)D(3) from kidney. As a result, CYP24 is an important multifunctional regulatory enzyme that maintains essential tissue-levels of Vitamin D hormone. As a part of our continuing interest in structure-function studies characterizing various binding proteins in the Vitamin D endocrine system, we targeted recombinant rat CYP24 with a radiolabeled 25-OH-D(3) affinity analog, and showed that the 25-OH-D(3)-binding site was specifically labeled by this analog. An affinity labeled sample of CYP24 was subjected to MS/MS analysis, which identified Ser57 as the only amino acid residue in the entire length of the protein that was covalently modified by this analog. Site-directed mutagenesis was conducted to validate the role of Ser57 towards substrate-binding. S57A mutant displayed significantly lower binding capacity for 25-OH-D(3) and 1,25(OH)(2)D(3). On the other hand, S57D mutant strongly enhanced binding for the substrates and conversion of 1,25(OH)(2)D(3) to calcitroic acid. The affinity probe was anchored via the 3-hydroxyl group of 25-OH-D(3). Therefore, these results suggested that the 3-hydroxyl group (of 25-OH-D(3) and 1,25(OH)(2)D(3)) in the S57D mutant could be stabilized by hydrogen bonding or a salt bridge leading to enhanced substrate affinity and metabolism.

White and black blood volumetric angiographic filtering: ellipsoidal scale-space approach.

Prefiltering is a critical step in three-dimensional (3-D) segmentation of the blood vessel and its display (see the recent book by Suri et al.). This paper presents a scale-space approach for filtering the white blood and black blood angiographic volumes and its implementation issues. The raw MR angiographic volume is first converted to isotropic volume followed by 3-D higher order separable Gaussian derivative convolution with known scales to generate edge volume. The edge volume is then run by the directional processor at each voxel where the eigenvalues of the 3-D ellipsoid are computed. The vessel score per voxel is then estimated based on these three eigenvalues which suppress the nonvasculature and background structures yielding the filtered volume. The filtered volume is ray-cast to generate the maximum intensity projection images for display. The performance of the system is evaluated by computing the mean, variance, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) images. The system is run over 20 patient studies from different areas of the body such as the brain, abdomen, kidney, knee, and ankle. The computer program takes around 150 s of processing time per study for a data size of 512 x 512 x 194, which includes the complete performance evaluation. We also compare our strategy with the recently published MR filtering algorithms by Alexander et al. and Sun et al.