Different Roles of Beclin1 in the Interaction Between Glia and Neurons after Exposure to Morphine and the HIV- Trans-Activator of Transcription (Tat) Protein.

Previously we showed that Beclin1 has a regulatory role in the secretion of inflammatory molecules in glia after exposure to morphine and Tat (an HIV protein). Here we show increased secretion of neuronal growth factors and increased neuronal survival in Beclin1-deficient glia. However, without glia co-culture, neurons deficient in Beclin1 showed greater death and enhanced dendritic beading when compared to wild-type neurons, suggesting that glial-secreted growth factors compensate for the damage reduced autophagy causes neurons. To assess if our ex vivo results correlated with in vivo studies, we used a wild-type (Becn1+/+) and Beclin1-deficient (Becn1+/+) mouse model and intracranially infused the mice with Tat and subcutaneously administered morphine pellets. After morphine implantation, significantly impaired locomotor activities were detected in both Becn1+/+ and Becn1+/- mice, irrespective of Tat infusion. After induction of pain, morphine-induced antinociception was detected. Interestingly, co-exposure to morphine and Tat increased sensitivity to pain in Becn1+/+ mice, but not in similarly treated Becn1+/- mice. Brain homogenates from Becn1+/+ mice exposed to Tat, alone and in combination with morphine, showed increased secretion of pro-inflammatory cytokines and reduced expression of growth factors when compared to similarly treated Becn1+/- mice. Likewise, increased neuronal loss was detected when both Tat and morphine were administered to Becn1+/+ mice, but not in similarly treated Becn1+/- mice. Overall, our findings show that there is a Beclin1-driven interaction between Tat and morphine in glia and neurons. Moreover, reduced glial-Beclin1 may provide a layer of protection to neurons under stressful conditions, such as when exposed to morphine and Tat.

Effect of paddy straw plus nonforage fiber sources based complete rations with different levels of neutral detergent fiber on hemato-biochemical and mineral profile of lactating dairy cows.

AIM: This study was conducted to assess the effect of feeding paddy straw plus nonforage fiber sources based complete rations with different levels of neutral detergent fiber (NDF) on hemato-biochemical and mineral parameters of lactating dairy cows. MATERIALS AND METHODS: The study was conducted for 6 months in 18 lactating dairy cows, divided into three groups of six each, by feeding them on paddy straw plus nonforage fiber sources based complete rations containing different levels of NDF, in two phases of 3 months (90 days) each, being the early and mid lactation phases, respectively. Three isonitrogenous and isocaloric complete rations, T1, T2 and T3 with 25%, 30% and 35% NDF, respectively, were fed to the experimental animals. Blood samples were collected at the beginning and the end of each of the two phases to estimate the different hematological, plasma protein, and mineral parameters to know the overall health status of the animals and standard methods were followed to analyze the samples. RESULTS: There was no significant difference (p>0.05) in various hematological parameters such as hemoglobin, glucose, and blood urea nitrogen (BUN) in blood; plasma protein parameters such as total protein, albumin, globulin and albumin: globulin ratio and mineral parameters such as plasma calcium and phosphorus levels at the beginning and end of Phase I (1st day and 90th day) and Phase II (91st day and 180th day) as well as between the three dietary treatments, with all the values being in the normal range for lactating dairy cows. Even though nonsignificant (p>0.05), the BUN values of animals fed on ration T1, both at the beginning and end of Phase I, were higher than that of animals fed on rations T2 and T3 because the diet T1 with lowest NDF and the highest soluble carbohydrate content underwent rapid fermentation in the rumen, produced more energy, which was utilized by the rumen microbes to degrade the protein in the feed to ammonia, the excess ammonia being transported to the liver and excreted through the blood resulting in a higher BUN content. CONCLUSION: Feeding of paddy straw plus nonforage fiber sources based complete rations with different levels of NDF had no effect on hemato-biochemical and mineral profile as well as overall health status of lactating dairy cows. However, the higher, BUN values found in cows fed on diet T1 with 25% NDF as compared to those fed on T2 and T3 with 30% and 35% NDF, respectively, indicate more wastage of protein in T1 as compared to T2 and T3, in early lactation.

Nucleotide diversity of Pita, a major blast resistance gene and identification of its minimal promoter.

Improvement of host plant resistance is one of the best methods to protect the yield from biotic stresses. Incorporation of major resistance genes or their variants into elite rice varieties will enhance the host plant resistance and its durability. Allele mining is a preferred choice to discover the novel allelic variants of major genes from wide range of germplasm. 'True' allele mining includes coding and noncoding regions, which are known to affect the plant phenotype, eventually. In this study, major blast resistance gene, Pita was analyzed by allele and promoter mining strategy and its different allelic variants were discovered from landraces and wild Oryza species. Polymorphisms at allelic sequences as well as transcription factor binding motif (TFBM) level were examined. At motif level, MYB1AT is present in Pita(Tadukan) and other resistance alleles, but was absent in the susceptible allele. Core promoter was demarked with 449 bp, employing serial promoter deletion strategy. Promoter with 1592 bp upstream region could express the gfp two fold higher than the core promoter. The identified Pita resistance allele (Pita(Konibora)) can be directly used in rice blast resistance breeding programs. Moreover, characterization of Pita core promoter led to deeper understanding of resistance gene's regulation and the identified core promoter can be utilized to express similar genes in rice.

Frequency and predictors for the risk of stroke recurrence up to 10 years after stroke: the South London Stroke Register.

BACKGROUND: Data estimating the risk of, and predictors for, long-term stroke recurrence are lacking. METHODS: Data were collected from the population-based South London Stroke Register. Patients were followed up for a maximum of 10 years. Kaplan-Meier estimates and Cox proportional hazards models were used to assess the cumulative risk of and predictors for first stroke recurrence. Variables analysed included sociodemographic factors, stroke subtype (defined as cerebral infarction, intracerebral haemorrhage and subarachnoid haemorrhage), stroke severity markers and prior-to-stroke risk factors. RESULTS: Between 1995 and 2004, 2874 patients with first-ever stroke were included. The mean follow-up period was 2.9 years. During 8311 person-years of follow-up, 303 recurrent events occurred. The cumulative risk of stroke recurrence at 1 year, 5 years and 10 years was 7.1%, 16.2% and 24.5% respectively. No differences in stroke recurrence were noted between the stroke subtypes. Factors increasing the risk of recurrence at 1 year were previous myocardial infarction (HR 1.73; 95% CI 1.08 to 2.78) and atrial fibrillation (HR 1.61; 95% CI 1.04 to 4.27); at 5 years, hypertension (HR 1.47; 95% CI 1.08 to 1.99) and atrial fibrillation (HR 1.79; 95% CI 1.29 to 2.49); and at 10 years, older age (p = 0.04), and hypertension (HR 1.38, 95% CI 1.04 to 1.82), myocardial infarction (HR 1.50, 95% CI 1.06 to 2.11) and atrial fibrillation (HR 1.51, 95% CI 1.09 to 2.09). CONCLUSIONS: Very-long-term risk of stroke recurrence is substantial. Different predictors for stroke recurrence were identified throughout the follow-up period. Risk factors prior to initial stroke have a significant role in predicting stroke recurrence up to 10 years.

Metabolic abnormalities in HIV type 1-infected children treated and not treated with protease inhibitors.

Our objective was to determine whether HIV-infected children treated with protease inhibitors (PIs) have different blood lipid, insulin, and glucose levels and body composition than HIV-infected children not treated with PIs. A cross-sectional cohort study was performed; in which 23 children were treated with combination antiretroviral therapy including a PI for at least 6 months and 12 children were treated with nucleoside reverse transcriptase inhibitors only (no-PI group). Levels of lipids, apolipoprotein B (apoB), insulin, and glucose were determined in the fasting state. Body composition and fat distribution were determined by anthropometric measurements and dual energy X-ray absorptiometry (DEXA) scan. Total cholesterol levels were higher in the PI-treated children (5.33 +/- 0.87 mM) than in the no-PI children (3.69 +/- 0.59 mM) (p < 0.0001). Similarly, low-density lipoprotein (LDL) levels were also elevated in the PI-treated children (3.27 +/- 0.76 vs. 2.14 +/- 0.51 mM) (p < 0.0001). ApoB and high-density lipoprotein (HDL), and to a lesser degree triglyceride levels, were also increased in the PI-treated children. Apart from percent arm fat as measured by DEXA, there were no differences between the two groups in measures of body composition or in their fasting glucose and insulin levels. The results from this cross-sectional cohort study suggest that the predominant lipid abnormalities associated with treatment with combination antiretroviral therapy including a PI in HIV-1-infected children are elevated total and LDL cholesterol.

Simple, sensitive, and specific detection of human immunodeficiency virus type 1 subtype B DNA in dried blood samples for diagnosis in infants in the field.

The detection of virus is used to diagnose human immunodeficiency virus type 1 (HIV-1) infection in infants due to the persistence of maternal antibodies for a year or more. An HIV-1 DNA PCR assay with simple specimen collection and processing was developed and evaluated. Whole blood was collected on filter paper that lysed cells and bound the DNA, eliminating specimen centrifugation and extraction procedures. The DNA remained bound to the filter paper during PCR amplification. Assays of copy number standards showed reproducible detection of 5 to 10 copies of HIV-1 in 5 microl of whole blood. The sensitivity of the assay did not decrease after storage of the standards on filter paper for 3 months at room temperature or after incubation at 37 or 45 degrees C for 20 h. The primers used for nested PCR of the HIV-1 pol gene amplified templates from a reference panel of multiple HIV-1 subtypes but did not amplify a subtype A or a subtype C virus from children living in Seattle. The assay had a sensitivity of 98.4% and a specificity of 98.3% for testing of 122 specimens from 35 HIV-1-infected and 16 uninfected children and 43 seronegative adults living in Washington. The assay had a sensitivity of 99% and a specificity of 100% for testing of 102 HIV-1-positive (as determined by enzyme immunoassay) Peruvian women and 6 seropositive and 34 seronegative infants. This assay, with adsorption of whole blood to filter paper and no specimen processing, provides a practical, economical, sensitive, and specific method for the diagnosis of HIV-1 subtype B infection in infants.

The effect of flicker rate on measured visual field extent in very young children.

PURPOSE: To investigate the effect of flicker rate on measured visual field extent in toddlers. METHODS: A total of 270 full-term children (90 each at 11-, 17-, and 30-months of age) and 36 adults were tested binocularly with an LED static perimetry procedure using a black double-arc perimeter. Each subject was tested with one of three flicker rates: 0, 3, or 10 Hz. The median farthest location seen and an interpolated estimate of the location at which 50% of the subjects detected the peripheral stimulus were calculated for each age group for each flicker rate. RESULTS: For 11-, 17-, and 30-month-old subjects, but not adults, flickering stimuli produced a larger measured visual field extent than nonflickering stimuli. For the 10-Hz stimuli, measured visual field extent in children did not differ from that of adults. CONCLUSIONS: In infants and young children, binocular measured visual field extent is enhanced by peripheral stimulus flicker. Maturity of the measured visual field depends on the stimulus parameters used during testing.

Oral ganciclovir in children: pharmacokinetics, safety, tolerance, and antiviral effects. The Pediatric AIDS Clinical Trials Group.

The pharmacokinetics, safety, tolerance, and antiviral effects of ganciclovir (Gcv) administered orally were evaluated in 36 children infected with cytomegalovirus (CMV) who were severely immunocompromised by infection with human immunodeficiency virus type 1. In this dose-escalation study, 30 mg/kg of Gcv administered every 8 h produced serum levels similar to the dose (1 g/8 h) effective for maintenance treatment of CMV retinitis in adults. In older children, serum Gcv concentrations were similar after the administration of capsules and suspension. All doses (10-50 mg/kg/8 h) studied were safe and, except for the volume of suspension or number of pills, were well tolerated. Oral Gcv was associated with a decrease in the detection of CMV by culture or polymerase chain reaction. CMV disease occurred in 3 children during the study: one developed Gcv resistance, another had harbored resistant virus at study entry, and a third had wild-type CMV

Inter-rater and intra-rater reliability in the interpretation of MTI Photoscreener photographs of Native American preschool children.

PURPOSE: To evaluate inter- and intra-rater reliability for the interpretation of MTI Photoscreener photographs taken in a population of Native American preschool children with a high prevalence of astigmatism. METHODS: Photographs of 369 children were rated by 11 nonexpert and 3 expert raters. Photographs for each child were scored as pass, refer, or retake. Nonexpert raters scored photos on two separate occasions, permitting analysis of intra-rater reliability. RESULTS: Analyses of pass/refer responses only: inter-rater reliability was moderate to substantial among nonexpert raters and substantial among expert raters. Intra-rater reliability among nonexperts was substantial. Analyses of all responses (pass, refer, and retake): inter-rater reliability for pass and refer scores was moderate among nonexperts and substantial among experts; for retake scores inter-rater reliability was slight for nonexperts and moderate for experts. Intra-rater reliability among nonexperts was substantial for pass and refer scores and moderate for retake scores. CONCLUSIONS: In this population with a high prevalence of astigmatism, whether MTI photoscreening results are interpretable is much more variable among and within raters than whether an interpretable photograph should be scored as pass or refer. The level of agreement among raters in the current study was influenced by the experience of the raters. In addition, nonexpert raters were more likely to deem a photograph uninterpretable than expert raters.

Stimulus motion increases measured visual field extent in children 3.5 to 30 months of age.

PURPOSE: To examine the influence of stimulus motion on measured visual field extent of 3.5- to 30-month-old children and adults. METHODS: Each subject was tested with LED-hybrid and LED-kinetic perimetry procedures, using a black double-arc perimeter. Targets in both procedures were identical in size, color, luminance, contrast, and flicker rate. However, in the LED-hybrid procedure, peripheral targets were sequentially illuminated from more peripheral to more central locations, whereas in the LED-kinetic procedure, a peripheral target on a black wand was manually moved centrally along the perimeter arm. A subset of subjects was also tested with white sphere kinetic perimetry (WSKP). RESULTS: The LED-kinetic procedure produced larger measured visual field extent than the LED-hybrid procedure in 3.5-, 11-, 17-, and 30-month-olds, but not in 7-month-olds or adults. Data from subjects tested with WSKP indicated that both stimulus motion and discrepancies in scoring methods contributed to the difference reported previously between visual field measurements obtained with WSKP vs. LED-hybrid perimetry. CONCLUSION: In infants and toddlers, measured visual field extent is larger for moving than for nonmoving targets. Further research is needed to determine whether the effect of motion is related to the visual system or to attentional factors.

HIV-1 dynamics in children.

HIV-1-infected children have higher plasma viral loads and progress to disease more quickly than infected adults. To gain insight into the accelerated pathogenesis of HIV-1 in children, viral dynamics were measured following the initiation of highly active antiretroviral therapy (HAART) and compared with those reported for adults. A biphasic decline in plasma HIV-1 RNA was observed, with a rapid decrease during the first 1 to 2 weeks of therapy (phase I) followed by a slower decline (phase II). The phase I and II decay rates were not significantly different among children of different ages, pretherapy plasma HIV-1 RNA levels, or CD4 cell counts. Estimated phase I decay rates were similar to those previously reported in adults with a mean of 0.43 days(-1) and a half-life of 1.6 days. The phase II decay rates were slower in children compared with adults with a mean of 0.016 days(-1) versus 0.066 days(-1), and a half-life of 43.3 versus 14.1 days, respectively (p < .05). The mean time required to reach viral levels below detection thresholds was also longer in these children compared with that in adults. These data suggest that HIV-1 dynamics may be different in children, and that these differences may necessitate different treatment strategies.

Does rate of stimulus presentation affect measured visual field extent in infants and toddlers?

PURPOSE: To evaluate the effect of stimulus presentation rate on the measurement of visual field extent in infants and toddlers. METHODS: Visual field extent was measured for 300 children (N = 60 at 3.5, 7, 11, 17, and 30 months) and 24 adults using hybrid static-kinetic perimetry. Flickering light-emitting diode (LED) stimuli were illuminated sequentially, peripherally to centrally at 10.2 degrees intervals, along 4 diagonal meridia at 2 stimulus presentation rates: 2 s/stimulus (equivalent to 5 degrees/s) and 3 s/stimulus (equivalent to 3 degrees/s). Rate of presentation was a between-subjects variable. RESULTS: No effect of stimulus presentation rate was found for adults. The faster rate of stimulus presentation yielded smaller measured visual field extent for children between the ages of 7 and 30 months. The apparent difference seen with 3.5-month-olds did not reach significance. CONCLUSIONS: Faster rates of stimulus presentation may result in underestimation of visual field extent in children between the ages of 7 and 30 months.

Risk of mother-to-infant transmission of HIV-1 is not reduced in CCR5/delta32ccr5 heterozygotes.

To determine if the 32-bp deletion of the chemokine receptor CCR5 (delta32ccr5) protects against mother-to-infant transmission of HIV-1, specimens from all uninfected and infected children who were perinatally exposed to HIV-1 and observed since 1988 and whose mothers did not take zidovudine were assessed for delta32ccr5. The CCR5 genotype was determined using polymerase chain reaction (PCR) for 122 subjects, of whom 73 were HIV-1 infected and 49 were perinatally exposed but uninfected; 70% and 71%, respectively, were Caucasian. Eleven of 73 (15%) infected children and 4 of 49 (8%) exposed uninfected children were CCR5/delta32ccr5 heterozygotes (p = 0.40). Among subjects who had at least one Caucasian parent or grandparent, 11 of 51 (22%) HIV-1-infected persons and 4 of 35 (11%) uninfected persons were heterozygotes. None were homozygous for the delta32ccr5 allele. The estimated relative risk for mother-to-infant HIV-1 transmission in heterozygotes was 2.0. Furthermore, the 95% confidence interval (0.6, 7.3) suggested that it is unlikely that the true relative risk was <0.6. Thus, the infant CCR5/delta32ccr5 heterozygous genotype was not associated with a diminished risk of perinatally acquired HIV-1 infection.

Clinical, virologic and immunologic responses of children with advanced human immunodeficiency virus type 1 disease treated with protease inhibitors.

OBJECTIVE: To determine the effects of combination antiretroviral therapy including a protease inhibitor (PI combination therapy) in children with advanced HIV-1 disease. STUDY DESIGN: An observational study of HIV-1 plasma RNA, lymphocyte subsets, delayed type hypersensitivity and physical growth after initiation of PI combination therapy. RESULTS: In nine children the median HIV-1 plasma RNA decreased 1.7 log10 (mean, 1.57; range, 0.7 to 2.2) following PI combination therapy and CD4 cells increased a median of 499 (mean, 528; range, 9 to 1088) cells/microl. A rebound of RNA, associated with the development of resistance to the PI, occurred in three subjects. Three of six children were no longer anergic and all nine achieved normal weight-growth velocities. Ritonavir was well-tolerated, despite its bitter taste; however, four of five children treated with indinavir developed renal complications. CONCLUSIONS: PI combination therapy in children with advanced HIV-1 disease was associated with a decrease in HIV-1 RNA, improved immunologic measures and normal or better weight gain. Of concern was the rebound in plasma HIV-1 associated with resistance to the PI observed in one-third of patients. This emphasizes the need for larger studies to define optimal PI containing regimens with long term efficacy in children.

Diminished interferon-gamma and lymphocyte proliferation in neonatal and postpartum primary herpes simplex virus infection.

Interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha production and lymphocyte proliferation in response to herpes simplex virus (HSV) antigen were assessed in 13 neonates and 3 parturient women with primary HSV infection. In comparison with 9 nonparturient adults, the neonates and parturient women showed significantly (P less than .01) diminished HSV antigen-stimulated lymphocyte proliferation and IFN-gamma production in the first 3-6 weeks after onset of infection. TNF alpha production did not differ significantly among HSV-infected groups. The impairment in neonatal cellular immunity was due, at least in part, to a specific deficit in response to HSV antigen. Lymphocyte proliferation and TNF alpha production in response to the mitogen concanavalin A (ConA) were comparable in adults and infants, but ConA-stimulated IFN-gamma production in infants was diminished throughout the study period. In contrast, HSV antigen-stimulated IFN-gamma production was comparable in infants and adults after 6 weeks. Not all patients with diminished cellular immune responses to HSV antigen manifested severe clinical disease. Nevertheless, patients with significant clinical morbidity had diminished cellular immune responses to HSV antigen. These results suggest that delayed acquisition of antigen-specific cellular immunity in primary HSV infection predisposes to more severe clinical disease.

Determination of submicrogram quantities of iron(III) by a catalytic polarographic method.

A simple and sensitive polarographic method has been developed for the determination of submicrogram quantities (0.01-0.16 mug/ml) of iron('III) based on the catalytic polarographic reduction of bromate in presence of resacetophenone isoniazid hydrazone. A 200-fold ratio of Mg(2+), Ca(2+), Cr(3+), Mn(2+), Co(2+), Ni(2+), Zn(2+), Al(3+) to Fe(3+) does not interfere. However, Cu(2+) and Cd(2+) interfere if they are present in more than 20-fold ratio to Fe(3+). Ti(4+), V(5+), Mo(6+) and W(6+) interfere even at 5-fold ratio. There is no interference from F(-), Cl(-), Br(-), I(-), SO(2-)(4), NO(-)(3) or NO(-)(2) even when present in large excess, but EDTA, CN(-), SCN(-), C(2)O(2-)(4), citrate and tartrate suppress the wave to a marked extent.