Admission hyperglycemia an independent predictor of outcome in acute ischemic stroke: A longitudinal study from a tertiary care hospital in South India.

BACKGROUND: Stroke is one of the important causes of long-term disability-related deaths worldwide. Incidence and prevalence of stroke have been steadily increasing in India. Increasing interest has been focused on the role of admission hyperglycemia in the evolution of acute ischemic stroke. Very few studies were conducted in south India; hence, we intended to do this study. METHODS: A total of 198 patients with acute ischemic stroke were included in a hospital-based longitudinal study to identify the independent factors (demographic, clinical, and biochemical parameters) associated with poor outcome (functional impairment-mRS ≥ 3 and mortality at 90 days of follow-up. RESULTS: Nearly, 75% of the patients presented with moderate-to-severe stroke. Out of the 198 patients, 117 (59%) had severe disability at admission. At 90 days of follow-up, it was observed that only 10 (5.5%) had severe disability. Patients with hyperglycemia exhibited greater functional impairment, that is, 96 out of 111 study subjects had modified Rankin score (mRS) ≥3, than those with normoglycemia (P < 0.0001). Mortality was high in hyperglycemics when compared with normoglycemics, that is, out of the 20 deaths, 13 patients had hyperglycemia and seven had normoglycemia at presentation (P < 0.015). Logistic regression analysis predicted that higher capillary blood glucose at first presentation, moderate-to-severe stroke, poor drug compliance, stress hyperglycemia, and newly detected diabetes mellitus were associated with poor functional outcome at 90 days of follow-up. CONCLUSION: Stress hyperglycemia in stroke was associated with higher risk of poor functional outcome in acute ischemic stroke. Hyperglycemia at stroke onset without prior history of diabetes mellitus have particularly poor prognosis, than those with hyperglycemia in known diabetes.

Midwives in India: a delayed cord clamping intervention using simulation.

BACKGROUND: Iron deficiency is a prevalent health problem in India affecting women and newborns. Delayed umbilical cord clamping at birth is a safe and effective means for increasing serum iron levels in newborns up to 6 months of age. AIM: The study aim was to increase the utilization of delayed cord clamping in a group of midwives working in Hyderabad, India. METHODS: A single group pre- and post-test design was used to evaluate knowledge, beliefs and practice before and after a delayed cord clamping intervention including follow-up at 10 months after the original intervention. The intervention included lectures and simulation. RESULTS: Results show significant increases in knowledge and positive beliefs about the practice of delayed cord clamping. Simulation was effective for eliciting important feedback related to learning. LIMITATIONS: Results represent a small group of midwives working with a non-profit foundation in Southern India. Language discordancy and cultural norms in this group of midwives may have influenced results. CONCLUSIONS: Knowledge, beliefs and practice related to delayed cord clamping were all significantly improved after the intervention. The Knowledge to Action framework using simulation is an effective cross-cultural method for implementing education about evidence-based practice. Midwives are invested in learning practices that promote public health. IMPLICATIONS FOR NURSING AND HEALTH POLICY: Changing institutional policy may have limitations without first considering normative practice. Using simulation combined with institutional health policy appears to result in significant uptake of practice change. Qualitative studies exploring the interconnections between cultural norms and decision making may be informative about promoting practice change particularly in this setting. Upscaling midwifery has been recommended to improve maternal and child health in India.

Management of airway and feeding difficulties in a neonate with Pierre Robin sequence: a case report.

Pierre Robin sequence is a congenital disorder that affects neonates, and is characterised by the classic triad of micrognathia, glossoptosis, and cleft of the secondary palate. We present the case of a child with Pierre Robin sequence who had severe respiratory distress and feeding problems, and describe a simple technique for forward traction of the mandible to relieve the respiratory distress and activate the tongue.

WRKY: its structure, evolutionary relationship, DNA-binding selectivity, role in stress tolerance and development of plants.

The plants during their sessile, autotrophic lifestyle are affected by wide range of environmental signals and regulate complex patterns of gene expression with the help of transcription factors. The WRKY transcription factors are considered as plant-specific, however, are also reported in protist, slime mold, fern and pine. The WRKY name is coined from its highly conserved 60 amino acid long WRKY domain. These TFs show W box specific binding which is also influenced by the W box flanking sequence. During evolution, the family has expanded in different patterns to facilitate distinct cellular, developmental, and physiological role in plants. The WRKY TFs form one of the largest families in flowering plants, and play a broad spectrum regulatory role as positive and negative regulators of plant defense regulation, abiotic stresses and also involved in growth and development of plants.

Effect of nickel on regeneration in Jatropha curcas L. and assessment of genotoxicity using RAPD markers.

The aim of the present study was to determine the effect of nickel on shoot regeneration in tissue culture as well as to identify polymorphisms induced in leaf explants exposed to nickel through random amplified polymorphic DNA (RAPD). In vitro leaf explants of Jatropha curcas were grown in nickel amended Murashige and Skoog (MS) medium at four different concentrations (0, 0.01, 0.1, 1 mM) for 3 weeks. Percent regeneration, number of shoots produced and genotoxic effects were evaluated by RAPD using leaf explants obtained from the first three treatments following 5 weeks of their subsequent subculture in metal free MS medium. Percent regeneration decreased with increase in addition of nickel to the medium up to 14 days from 42.31% in control to zero in 1.0 mM. The number of shoot buds scored after 5 weeks was higher in control as compared to all other treatments except in one of the metal free subculture medium wherein the shoot number was higher in 0.01 mM treatment (mean = 7.80) than control (mean = 7.60). RAPD analysis produced only 5 polymorphic bands (3.225%) out of a total of 155 bands from 18 selected primers. Only three primers OPK-19, OPP-2, OPN-08 produced polymorphic bands. The dendrogram showed three groups A, B, and C. Group A samples showed 100% genetic similarity within them. Samples between groups B and C were more genetically distant from each other as compared to samples between groups A and B as well as groups A and C. Cluster analysis based on RAPD data correlated with treatments.

Development and validation of high-throughput liquid chromatography-tandem mass spectrometric method for simultaneous quantification of clopidogrel and its metabolite in human plasma.

A simple, sensitive and reliable method is described for simultaneous quantification of Clopidogrel and its metabolite in human plasma by using HTLC-MS/MS. The analytical procedure involves on-line coupling of extraction with Cyclone P (50 mm x 0.5 mm 50 microm) HTLC column by injecting 15 microL sample and chromatographic separation is performed with Cohesive Propel C18 (5 microm, 3.0 mm x 50 mm), followed by quantification with mass detector in SRM mode using ESI as an interface. The calibration curves were linear over a concentration range of 0.1-8 ng/mL of Clopidogrel and 70 ng/mL to 6 microg/mL of its metabolite using 20 mL human plasma per batch. The total run time of analysis was 7.5 min and the lower limits of quantification were 0.1 ng/mL for Clopidogrel and 70 ng/mL for its metabolite. The method validation was carried out in terms of specificity, sensitivity, linearity, precision, accuracy and stability. The validated method was applied in bioavailability and bioequivalence study.

Spectrophotometric estimation of bromide ion in excess chloride media.

The redox reaction between bromate and chloride ions in the presence and the absence of two or less equivalents of bromide ion ascertaining the formation of bromine chloride species of type BrCl and BrCl(2)(-) in subsequent reactions in 4% H(2)SO(4), has been studied by spectrophotometry. Calibration graphs for the bromide ion estimation in 0.1% KBrO(3)-4% H(2)SO(4) medium are determined separately in the presence of known amounts of NaCl. The effect of Cl(-) ion percentage on the determination of Br(-) ion is studied and reported herewith a suitable equation for a precise, reliable and quick spectrophotometric estimation.

Modification of the Fc region of a primatized IgG antibody to human CD4 retains its ability to modulate CD4 receptors but does not deplete CD4(+) T cells in chimpanzees.

Keliximab, a Primatized IgG1 CD4 mAb, was reconfigured to an IgG4 antibody. The gamma4 constant region was further modified by substituting glutamic acid for serine at position 235 in the CH2 domain (IgG4-E), to remove residual binding to Fcgamma receptors, and substitution of serine with proline at position 228 in the hinge region (IgG4-PE) for greater stability. Pharmacokinetic analysis in rats gave a t(1/2) of approximately 4 days for IgG4-E and 9 days for IgG4-PE, consistent with a greater stability of the IgG4-PE molecule. The effects on T cell subsets were assessed in chimpanzees given escalating doses of IgG4-PE: 0.05 mg/kg on Day 16, 1.5 mg/kg dose on Day 43, and 15 mg/kg on Day 85. Receptor modulation was observed at the two highest doses, but no depletion of T cells at any dose. The in vitro and in vivo results demonstrate the potential of this IgG4-PE mAb for use in human trials.

Temperature-sensitive differential affinity of TRAIL for its receptors. DR5 is the highest affinity receptor.

TRAIL is a member of the tumor necrosis factor (TNF) family of cytokines which induces apoptotic cell death in a variety of tumor cell lines. It mediates its apoptotic effects through one of two receptors, DR4 and DR5, which are members of of the TNF receptor family, and whose cytoplasmic regions contain death domains. In addition, TRAIL also binds to 3 "decoy" receptors, DcR2, a receptor with a truncated death domain, DcR1, a glycosylphosphatidylinositol-anchored receptor, and OPG a secreted protein which is also known to bind to another member of the TNF family, RANKL. However, although apoptosis depends on the expression of one or both of the death domain containing receptors DR4 and/or DR5, resistance to TRAIL-induced apoptosis does not correlate with the expression of the "decoy" receptors. Previously, TRAIL has been described to bind to all its receptors with equivalent high affinities. In the present work, we show, by isothermal titration calorimetry and competitive enzyme-linked immunosorbent assay, that the rank order of affinities of TRAIL for the recombinant soluble forms of its receptors is strongly temperature dependent. Although DR4, DR5, DcR1, and OPG show similar affinities for TRAIL at 4 degrees C, their rank-ordered affinities are substantially different at 37 degrees C, with DR5 having the highest affinity (K(D)

Elimination of Fc receptor-dependent effector functions of a modified IgG4 monoclonal antibody to human CD4.

Several CD4 mAbs have entered the clinic for the treatment of autoimmune diseases or transplant rejection. Most of these mAbs caused CD4 cell depletion, and some were murine mAbs which were further hampered by human anti-mouse Ab responses. To obviate these concerns, a primatized CD4 mAb, clenoliximab, was generated by fusing the V domains of a cynomolgus macaque mAb to human constant regions. The heavy chain constant region is a modified IgG4 containing two single residue substitutions designed to ablate residual Fc receptor binding activity and to stabilize heavy chain dimer formation. This study compares and contrasts the in vitro properties of clenoliximab with its matched IgG1 derivative, keliximab, which shares the same variable regions. Both mAbs show potent inhibition of in vitro T cell responses, lack of binding to complement component C1q, and inability to mediate complement-dependent cytotoxicity. However, clenoliximab shows markedly reduced binding to Fc receptors and therefore does not mediate Ab-dependent cell-mediated cytotoxicity or modulation/loss of CD4 from the surface of T cells, except in the presence of rheumatoid factor or activated monocytes. Thus, clenoliximab retains the key immunomodulatory attributes of keliximab without the liability of strong Fcgamma receptor binding. In initial clinical trials, these properties have translated to a reduced incidence of CD4+ T cell depletion.

Stress signaling through Ca2+/calmodulin-dependent protein phosphatase calcineurin mediates salt adaptation in plants.

Calcineurin (CaN) is a Ca2+- and calmodulin-dependent protein phosphatase (PP2B) that, in yeast, is an integral intermediate of a salt-stress signal transduction pathway that effects NaCl tolerance through the regulation of Na+ influx and efflux. A truncated form of the catalytic subunit and the regulatory subunit of yeast CaN were coexpressed in transgenic tobacco plants to reconstitute a constitutively activated phosphatase in vivo. Several different transgenic lines that expressed activated CaN also exhibited substantial NaCl tolerance, and this trait was linked to the genetic inheritance of the CaN transgenes. Enhanced capacity of plants expressing CaN to survive NaCl shock was similar when evaluation was conducted on seedlings in tissue culture raft vessels or plants in hydroponic culture that were transpiring actively. Root growth was less perturbed than shoot growth by NaCl in plants expressing CaN. Also, NaCl stress survival of control shoots was enhanced substantially when grafted onto roots of plants expressing CaN, further implicating a significant function of the phosphatase in the preservation of root integrity during salt shock. Together, these results indicate that in plants, like in yeast, a Ca2+- and calmodulin-dependent CaN signal pathway regulates determinants of salt tolerance required for stress adaptation. Furthermore, modulation of this pathway by expression of an activated regulatory intermediate substantially enhanced salt tolerance.

After a stroke: strategies to restore function and prevent complications.

Mortality and morbidity are high in elderly stroke patients. Early mobilization and prevention of stroke-related complications improve their ability to participate in a more intense and comprehensive rehabilitation program. An interdisciplinary approach to stroke rehabilitation restores functional loss, improves quality of life, and decreases long-term economic costs. Important parts of stroke rehabilitation include patient and family education, treatment of stroke-related complications, and prevention of recurrent stroke. A healthy and caring spouse, continence of bladder and bowel, and ability to feed oneself are the most positive predictors of stroke outcome.

Stroke rehabilitation.

Stroke is the leading cause of brain damage and resultant disability. Rehabilitation measures help to restore lost abilities, improve quality of life and decrease the long-term economic cost of stroke. Proper patient selection, realistic goal setting, the active participation of both the patient and family, and the use of an interdisciplinary team approach are important for the success of stroke rehabilitation. Functional demand and intensive training are believed to trigger central nervous system reorganization, which is responsible for late functional recovery after stroke. The outcome following a stroke is most likely to be positive when patients have bladder and bowel continence, are able to feed themselves and have a healthy and caring spouse. Stroke rehabilitation must include the prevention or early diagnosis of medical complications as well as patient and family education concerning the prevention of recurrent stroke.

Pyridinyl imidazoles inhibit the inflammatory phase of delayed type hypersensitivity reactions without affecting T-dependent immune responses.

The effects of pyridinyl imidazoles, specifically SK&F 105809 and its metabolite, on normal T-cell and B-cell mediated immune responses were examined and compared to the fungal macrolide immunosuppressives, cyclosporin A, FK506 and rapamycin and to the corticosteroid, dexamethasone. The orally active prodrug SK&F 105809 [2-(4-methylsulfinylphenyl)-3-(4-pyridyl)-6,7-dihydro-[5H]-pyrrolo [1,2-a] imidazole[ and its metabolite, SK&F 105561 [2-(4-methylthiophenyl)-3-(4-pyridyl)-6,7-dihydro-[5H]-pyrrolo[1,2 -a] imidazole] are dual 5-lipoxygenase (5-LO) and cycloxygenase (CO) inhibitors with potent anti-inflammatory and cytokine (IL-1/TNF) suppressive activities. The anti-inflammatory activity of SK&F 105809 and its metabolite were evaluated in an antigen-specific murine model of delayed type hypersensitivity (DTH) response, where they were found to affect only the inflammatory and not the induction phase of this response. In contrast, these compounds and other pyridinyl imidazoles (SK&F 86002 and its analog, SK&F 104351) exhibited no immunosuppressive activity under conditions where the macrolide rapamycin and the corticosteroid dexamethasone abrogated both the cellular and humoral immune responses. Thus, the ability of pyridinyl imidazoles to attenuate independently the inflammatory components of the disease without causing generalized immunosuppression enhances their profile as candidates for therapy of chronic inflammatory diseases, specifically those mediated by cytokines (e.g. IL-1, TNF) and eicosanoids.

A novel 500,000 Da, linear, single chain extracellular protein synthesized by several childhood tumors.

A high molecular weight extracellular protein has been purified from cell culture medium of Ewing's sarcoma cell lines, by high performance liquid chromatography and electroelution from SDS-PAGE electrophoresis. This protein has an apparent molecular mass of about 500,000 Da on SDS-PAGE. Immunoprecipitation studies with several extracellular matrix glycoproteins (laminin, fibronectin) specific antisera indicate it is a separate protein. Reduction of disulphide bonds with 2-ME or DTT fails to significantly alter its migration on SDS-PAGE gels, other than a slight apparent increase in molecular mass, indicating an apparent single polypeptide chain structure. The slightly greater mobility observed in unreduced gels suggests one or more regions of intrachain disulfide bonding. It is sensitive to pepsin and trypsin, but resistant to bacterial collagenase indicating that it does not contain collagenous domains. Metabolic labelling with 3H-proline, 3H-leucine, and 35S-methionine indicate that this protein is proline-poor, but leucine, and especially methionine, rich. Sodium 35S-sulfate incorporation is totally negative and treatment with glycosaminoglycan degrading enzymes has no effect on the mobility of the protein on gels, unlike typical proteoglycans. This protein appears by rotary shadowing electron microscopy as a long, thin, filamentous molecule at least 500 nm (0.5 um) in length. The tissue localization and function are unknown at this time, but are under active investigation.

Inhibition of vesicular stomatitis virus protein synthesis and infection by sequence-specific oligodeoxyribonucleoside methylphosphonates.

Oligodeoxyribonucleoside methylphosphonates which have sequences complementary to the initiation codon regions of N, NS, and G vesicular stomatitis virus (VSV) mRNAs were tested for their ability to inhibit translation of VSV mRNA in a cell-free system and in VSV-infected mouse L cells. In a rabbit reticulocyte lysate cell-free system, the oligomers complementary to N (oligomer I) and NS (oligomer II) mRNAs inhibited translation of VSV N and NS mRNAs whereas oligomer III had only a slight inhibitory effect on N protein synthesis. At 100 and 150 microM, oligomer I specifically inhibited N protein synthesis in the lysate. In contrast, at 150 microM, oligomer II inhibited both N and NS protein synthesis. This reduced specificity of inhibition may be due to the formation of partial duplexes between oligomer II and VSV N mRNA. The oligomers had little or no inhibitory effects on the synthesis of globin mRNA in the same lysate system. Oligomers I-III specifically inhibited the synthesis of all five viral proteins in VSV-infected cells in a concentration-dependent manner. The oligomers had no effects on cellular protein synthesis in uninfected cells nor on cell growth. An oligothymidylate which forms only weak duplexes with poly(rA) had just a slight effect on VSV protein synthesis and yield of virus. Oligomers I-III have extensive partial complementarity with the coding regions of L mRNA. The nonspecific inhibition of viral protein synthesis in infected cells may reflect the role of N, NS, and/or L proteins in the replication and transcription of viral RNA or result from duplex formation between the oligomers and complementary, plus-strand viral RNA.(ABSTRACT TRUNCATED AT 250 WORDS)

A guide to early mobilization of bedridden elderly.

A vigorous, functionally oriented program should be started as early as the patient's medical or surgical condition permits. This comprehensive approach requires the efforts of a team of health professionals coordinated by the patient's primary care physician. Restoration of strength, flexibility, coordination, and balance are the goals of successful ambulation. When an elderly person has difficulty in walking, the physician and therapist must search for the cause and take corrective measures.

Solid-phase syntheses of oligodeoxyribonucleoside methylphosphonates.

Oligodeoxyribonucleoside methylphosphonates of defined sequence of the type d-Np(NP)nN, where n is 6-13, are readily prepared on insoluble polystyrene supports by use of protected 5'-(dimethoxytrityl)deoxyribonucleoside 3'-(methylphosphonic imidazolides) as synthetic intermediates. The imidazolides are prepared in situ by reaction of protected 5'-(dimethoxytrityl)deoxyribonucleoside with methylphosphonic bis(imidazolide) and can be stores in the reaction solution for up to 2 weeks at 4 degrees C with no loss in activity. The condensation reaction is accelerated by the presence of tetrazole, which appears to act as an acid catalyst. The half-life for dimer formation on the polystyrene support is 5 min, and the reaction is 95% complete after 60 min. Although similar kinetics are observed when controlled pore glass is used as the support, the extent of the reaction does not go beyond 78%, even after prolonged incubation. In order to simplify purification and sequence analysis of the oligomer, the 5'-terminal nucleoside unit is linked via a phosphodiester bond. This linkage may be introduced by either an o-chlorophenyl phosphotriester method or a cyanoethyl phosphoramidite method. The latter procedure simplifies the deprotection step, since the cyanoethyl group is readily cleaved by ethylenediamine, which also removes the base protecting groups and cleaves the oligomer from the support. The singly charged oligomers are easily purified by affinity chromatography on DEAE-cellulose. The chain lengths of the oligomers were confirmed after 5'-end labeling with polynucleotide kinase by partial hydrolysis of the methylphosphonate linkages with 1 M aqueous piperidine followed by polyacrylamide gel electrophoresis of the hydrolysate.(ABSTRACT TRUNCATED AT 250 WORDS)

Antiviral effect of an oligo(nucleoside methylphosphonate) complementary to the splice junction of herpes simplex virus type 1 immediate early pre-mRNAs 4 and 5.

Selective inhibition of regulatory immediate early (IE) genes of herpes simplex virus type 1 (HSV-1) should inhibit virus growth. Treatment of HSV-1-infected cells with the oligo(nucleoside methylphosphonate) d(TpCCTCCTG) (deoxynucleoside methylphosphonate residues in italic), which is complementary to the acceptor splice junction of HSV-1 IE pre-mRNA 4 and 5, before (1-24 hr) or at the time of infection caused a dose-dependent inhibition in virus replication. Virus titers were decreased 50% and 90% in cells treated with 25 microM and 75 microM oligomer, respectively; at 300 microM, a 99% reduction in virus production was observed. Viral DNA synthesis was reduced 70-75% and there was a 90% reduction in synthesis of viral proteins, including other IE species and viral functional (130-kDa major DNA-binding) and structural (glycoprotein gB) proteins. In the same concentration range, d(TpCCTCCTG) caused a minimal reduction (0-30%) in protein synthesis and growth rates (less than 40%) of uninfected cells. The data suggest that oligo(nucleoside methylphosphonate)s may be effective in antiviral chemotherapy.