Evaluation of memory B cells among perinatally HIV infected children from Houston Texas

Background: HIV associated B cell exhaustion is a notable characteristic of HIV viremic adults. However, it is not known if such alterations are present in perinatal HIV infected children, whose viral dynamics differs from those seen in adults. In the present study we perform an analysis of B cells subsets and measure antigen-specific memory B cells (MBC) in a pediatric HIV infected cohort. ^ Methods: Peripheral mononuclear cells (PBMC) of perinatal HIV infected individuals are characterized into naïve (CD21hi/CD27−), classic (CD27+), tissue like (CD21lo/CD27 −) and activated MBC (CD27+CD21− ) by FACS. A memory ELISPOT assay is used to detect antibody secreting cells. We measure total IgG and antibodies specific for influenza, HBV, mumps, measles, rubella and VZV. Memory was expressed as spot forming cells (SPC) /million of PBMC. Wilcoxon rank-sum was used to compare unpaired groups and linear regression analysis was used to determine predictors of B cell dysfunction ^ Results: 41 HIV perinatal infected children are included (51.2% females and 65.9% Black). Age at study is median (range) 8.78 years (4.39-11.57). At the time of testing they have a CD4% of 30.9 (23.2-39.4), a viral load (VL) of 1.95 log10 copies/ml (1.68-3.29) and a cumulative VL of 3.4 log10 copy × days (2.7-4.0). Ninety two percent of the children are on cARV for > 6 months. Overall, HIV+ children compared with controls have a significant lower number of IgG and antigen specific SFC. In addition, they have a lower proportion of classical MBC 12.9 (8.09-19.85) vs 29.4 (18.7-39.05); 0.01, but a significant higher proportion of tissue like memory MBC 6.01 (2.79-12.7) vs 0.99 (0.87-1.38); 0.003, compared with controls. Patients are parsed on VL (<400 and ≥ 400 copies/ml) with the objective to evaluate the effect of VL on B cell status. Patients with a VL ≥ 400 copies/ml have a significantly lower IgG, HBV, measles, rubella and VZV SPC compared with those with a VL < 400 copies/ml. There are no significant differences in B cell subpopulations between the groups. A moderate negative correlation was observed between the time of cARV initiation and the frequency of IgG memory B cells, suggesting that early initiation of cARV appears to lead to a better functionality of the IgG memory B cells (P=0.05). A statistically significant positive correlation was observed between the total number of IgG memory cells and the number of antigen-specific memory B cells/SPCs. Suggesting that the progressive recovery of the IgG memory B cell pull goes along with a progressive increase in the number of antigen-specific SPCs. ^ Conclusion: A pediatric cohort in overall good status with respect to HIV infection and on ART has defects in B cell function and numbers (reduced total and antigen specific MBC and increased tissue like and reduced classical MBC).^

Epiphytic orchid diversity found on trees and shrubs in coffee cultivated Afromontane rainforest, SW Ethiopia

The moist evergreen Afromontane forest of SW Ethiopia has become extremely fragmented and most remnants are intensively managed for cultivation of coffee (Coffea arabica). We investigated the distributions of epiphytic orchids in shade trees and their understory in forests with contrasting management intensity to determine biodiversity losses associated with coffee cultivation and to determine the capacity of coffee shrubs to act as refugia for orchid species. We studied epiphytic orchids in managed forests and natural forests and recorded orchid diversity and abundance in different tree zones of 339 trees and in the understory. Coffee management was associated with a downward shift of orchid species as orchid species were occurring in significantly lower tree zones in managed forest. The number of shrubs in the understory of managed forest was not higher than in natural forests, yet orchid abundance was higher in the understory of managed forests. Local extinctions of epiphytic orchids and species losses in the outer tree zones (a contraction of habitat) in managed forests are most likely driven by losses of large, complex-structured climax trees, and changes in microclimate, respectively. Coffee shrubs and their shade trees in managed forests are shown here to be a suitable habitat for only a limited set of orchid species. As farmers continue to convert natural forest into managed forest for coffee cultivation, further losses of habitat quality and collateral declines in regional epiphytic orchid diversity can be expected. Therefore, the conservation of epiphytic orchid diversity, as well as other components of diversity of the coffee forests, must primarily rely on avoiding coffee management intensification in the remaining natural forest. Convincing farmers to keep forest-climax trees in their coffee forest and to tolerate orchids on their coffee shrubs may also contribute to a more favorable conservation status of orchids in Ethiopian coffee agroecosystems.

Ensayos acelerados de fiabilidad de células solares de concentración

SUMMARY Concentration Photovoltaic Systems (CPV) have been proposed as an alternative to conventional systems. During the last years, there has been a boom of the CPV industry caused by the technological progress in all the elements of the system. and mainly caused by the use of multijunction solar cells based on III-V semiconductors, with efficiencies exceeding to 43%. III-V solar cells have been used with high reliability results in a great number of space missions without concentration. However, there are no previous results regarding their reliability in concentration terrestrial applications, where the working conditions are completely different. This lack of experience, together with the important industrial interest, has generated the need to evaluate the reliability of the cells. For this reason, nowadays there are several research centers around the undertaking this task. The evaluation of the reliability of this type of devices by means of accelerated tests is especially problematic when they work at medium or high concentration, because it is practically impossible to emulate real working conditions of the cell inside climatic chambers. In fact, as far as we know, the results that appear in this Thesis are the first estimating the Activation Energy of the failure mechanism involved, as well as the warranty of the III-V concentrator solar cells tested here. To evaluate the reliability of III-V very high concentrator solar cells by means of accelerated tests, a variety of activities, described in this Thesis have been carried out. The First Part of the memory presents the theoretical part of the Doctoral Thesis. After the Introduction, chapter 2 presents the state of the art in degradation and reliability of CPV systems and solar cells. Chapter 3 introduces some reliability definitions and the application of specific statistical functions to the evaluation of the reliability and parameters. From these functions, important parameters will be calculated to be used later in the experimental results of Thesis. The Second Part of the memory contains the experimental. Chapter 4 shows the types of accelerated tests and the main goals pursuit with them when carried out over CPV systems and solar cells. In order to evaluate quantitatively the reliability of the III-V concentrator solar cells used in these tests, some modifications have been introduced which discussion will be tackled here. Based on this analysis the working plan of the tests carried out in this Doctoral Thesis is presented. Chapter 5 presents a new methodology as well as the necessary instrumentation to carry out the tests described here. This new methodology takes into account the adaptation, improvement and novel techniques needed to test concentrator solar cells. The core of this memory is chapter 6, which presents the results of the characterization of the cells during the accelerated life tests and the analysis of the aforementioned results with the purpose of getting quantitative values of reliability in real working conditions. The acceleration factor of the accelerated life tests, under nominal working conditions has been calculated. Accordingly, the validity of the methodology as well as the calculations based on the reliability assessment, have also been demonstrated. Finally, quantitative values of degradation, reliability and warranty of the solar cells under field nominal working conditions have been calculated. With the development of this Doctoral Thesis the reliability of very high concentrator GaAs solar cells of small area has been evaluated. It is very interesting to generalize the procedures described up to this point to III-V multijunction solar cells of greater area. Therefore, chapter 7 develops this generalization and introduces also a useful thermal modeling by means of finite elements of the test cells’ circuits. In the last chapter, the summary of the results and the main contributions of this Thesis are outlined and future research activities are identified. RESUMEN Los Sistemas Fotovoltaicos de Concentración (SFC) han sido propuestos como una alternativa a los sistemas convencionales de generación de energía. Durante los últimos años ha habido un auge de los SFC debido a las mejoras tecnológicas en todos los elementos del sistema, y principalmente por el uso de células multiunión III-V que superan el 43% de rendimiento. Las células solares III-V han sido utilizadas con elevada fiabilidad en aplicaciones espaciales sin concentración, pero no existe experiencia de su fiabilidad en ambiente terrestre a altos niveles de concentración solar. Esta falta de experiencia junto al gran interés industrial ha generado la necesidad de evaluar la fiabilidad de las células, y actualmente hay un significativo número de centros de investigación trabajando en esta área. La evaluación de la fiabilidad de este tipo de dispositivos mediante ensayos acelerados es especialmente problemática cuando trabajan a media o alta concentración por la casi imposibilidad de emular las condiciones de trabajo reales de la célula dentro de cámaras climáticas. De hecho, que sepamos, en los resultados de esta Tesis se evalúa por primera vez la Energía de Activación del mecanismo de fallo de las células, así como la garantía en campo de las células de concentración III-V analizadas. Para evaluar la fiabilidad de células solares III-V de muy alta concentración mediante ensayos de vida acelerada se han realizado diversas actividades que han sido descritas en la memoria de la Tesis. En la Primera Parte de la memoria se presenta la parte teórica de la Tesis Doctoral. Tras la Introducción, en el capítulo 2 se muestra el estado del arte en degradación y fiabilidad de células y Sistemas Fotovoltaicos de Concentración. En el capítulo 3 se exponen de forma resumida las definiciones de fiabilidad y funciones estadísticas que se utilizan para la evaluación de la fiabilidad y sus parámetros, las cuales se emplearán posteriormente en los ensayos descritos en este Tesis. La Segunda Parte de la memoria es experimental. En el capítulo 4 se describen los tipos y objetivos de los ensayos acelerados actualmente aplicados a SFC y a las células, así como las modificaciones necesarias que permitan evaluar cuantitativamente la fiabilidad de las células solares de concentración III-V. En base a este análisis se presenta la planificación de los trabajos realizados en esta Tesis Doctoral. A partir de esta planificación y debido a la necesidad de adaptar, mejorar e innovar las técnicas de ensayos de vida acelerada para una adecuada aplicación a este tipo de dispositivos, en el capítulo 5 se muestra la metodología empleada y la instrumentación necesaria para realizar los ensayos de esta Tesis Doctoral. El núcleo de la memoria es el capítulo 6, en él se presentan los resultados de caracterización de las células durante los ensayos de vida acelerada y el análisis de dichos resultados con el objetivo de obtener valores cuantitativos de fiabilidad en condiciones reales de trabajo. Se calcula el Factor de Aceleración de los ensayos acelerados con respecto a las condiciones nominales de funcionamiento a partir de la Energía de Activación obtenida, y se demuestra la validez de la metodología y cálculos empleados, que son la base de la evaluación de la fiabilidad. Finalmente se calculan valores cuantitativos de degradación, fiabilidad y garantía de las células en condiciones nominales en campo durante toda la vida de la célula. Con el desarrollo de esta Tesis Doctoral se ha evaluado la fiabilidad de células III-V de área pequeña, pero es muy interesante generalizar los procedimientos aquí desarrollados para las células III-V comerciales de área grande. Por este motivo, en el capítulo 7 se analiza dicha generalización, incluyendo el modelado térmico mediante elementos finitos de los circuitos de ensayo de las células. En el último capítulo se realiza un resume del trabajo y las aportaciones realizadas, y se identifican las líneas de trabajo a emprender en el futuro.

Passenger-car equivalents of trucks in composite traffic. Final report.

Federal Highway Administration, Office of Program and Policy Planning, Washington, D.C.

Preliminary human factors guidelines for automated highway system designers. Volume I: guidelines for AHS designers.

Federal Highway Administration, Washington, D.C.

Assessment of the benefits of two-way center left turn lanes. Phase I: report (revised).

Michigan Department of Transportation, Lansing

Planning for court monitoring. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Emergency care systems demonstration projects. Volume 2: the emergency care system. Final report.

National Highway Safety Bureau, Washington, D.C.

Operational and geometric evaluation of exclusive truck lanes. Final report.

Texas State Department of Highways and Public Transportation, Transportation Planning Division, Austin

Identification and assessment of organizations that would distribute traffic safety information to older drivers and pedestrians. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Evaluation of rail rapid transit and express bus service in the urban commuter market. Final report.

Transportation Department, Office of Transportation Planning Analysis, Washington, D.C.

The application of proteomics to the human alcoholic brain

Alcoholism results in changes in the human brain that reinforce the cycle of craving and dependency, and these changes are manifest in the pattern of expression of proteins in key cells and brain areas. Described here is a proteomics-based approach aimed at determining the identity of proteins in the superior frontal cortex (SFC) of the human brain that show different levels of expression in autopsy samples taken from healthy and long-term alcohol abuse subjects. Soluble protein fractions constituting pooled samples combined from SFC biopsies of four well-characterized chronic alcoholics (mean consumption > 80 g ethanol/day throughout adulthood) and four matched controls (< 20 g/day) were generated. Two-dimensional electrophoresis was performed in triplicate on alcoholic and control samples and the resultant protein profiles analyzed for differential expression. Overall, 182 proteins differed by the criterion of twofold or more between case and control samples. Of these, 139 showed significantly lower expression in alcoholics, 35 showed significantly higher expression, and 8 were new or had disappeared. To date, 63 proteins have been identified using MALDI-MS and MS-MS. The finding that the expression level of differentially expressed proteins is preponderantly lower in the alcoholic brain is supported by recent results from parallel studies using microarray mRNA transcript.

Genes and gene expression in the brains of human alcoholics

Chronic alcohol misuse by human subjects leads to neuronal loss in regions such as the superior frontal cortex (SFC). Propensity to alcoholism is associated with several genes. γ-Aminobutyric acid (GABA)A receptor expression differs between alcoholics and controls, whereas glutamate receptor differences are muted. We determined whether genotype differentiated the regional presentation of GABAA and glutamate-NMDA (N-methyl-d-aspartate) receptors in SFC. Autopsy tissue was obtained from alcoholics without comorbid disease, alcoholics with liver cirrhosis, and matched controls. ADH1C, DRD2B, EAAT2, and APOE genotypes modulated GABAA-β subunit protein expression in SFC toward a less-effective form of the receptor. Most genotypes did not divide alcoholics and controls on glutamate-NMDA receptor pharmacology, although gender and cirrhosis did. Genotype may affect amino acid transmission locally to influence neuronal vulnerability.

Comorbidity and genotype modify receptor expression in human alcoholics

Chronic alcohol misuse leads to both widespread and localized damage in human cerebral cortex. The latter, as neuronal loss, is marked in superior frontal cortex (SFC) but milder in primary motor cortex (PMC) and elsewhere. Quantitative morphometry by Harper et al showed that neuronal loss is greater in alcoholics with comorbidity (Wernicke Korsakoff syndrome, liver cirrhosis). Previous work revealed a paradox: the marked differences in GABAA receptor density, pharmacology, and expression between alcoholics without cormorbidity and controls are muted or absent in cirrhotic alcoholics. This concurs with work by the Butterworth group on hepatic encephalopathy cases — most of whom had an alcoholic ætiology — who show only minor differences from controls. Glutamate receptor differences are muted in many autopsy studies, though we have evidence that NMDA site pharmacology may vary in cirrhotic alcoholics. Here we used Real-Time PCR normalized to GAPDH deltaCT to quantify NMDA NR1, NR2A and NR2B subunit expression in SFC and PMC samples obtained at autopsy from alcoholics with and without comorbid cirrhosis and matched controls. Overall subunit transcript expression was signifi cantly lower in alcoholic cirrhotics than in either of the other groups (F2,42 = 12.942, P < 0.001). The effect was most marked for the NR1 subunit; males differed from females, particularly in SFC. The data suggest that if excitotoxicity mediates neuronal loss in SFC, it may be implemented differently: passively in uncomplicated alcoholics, by altered GABAergic transmission; actively in cirrhotic alcoholics, by altered glutamatergic transmission. We also subdivided cases on a panel of genetic markers. Different genotypes interacted with NMDA and GABAA pharmacology and expression. Cirrhotic and uncomplicated alcoholics may differ pathogenically because of inherent characteristics in addition to possible neurotoxic sequelæ to the liver damage.

Microarray and proteomic analysis of the human alcoholic brain

The superior frontal cortex (SFC) is selectively damaged in chronic alcohol abuse, with localized neuronal loss and tissue atrophy. Regions such as motor cortex show little neuronal loss except in severe co-morbidity (liver cirrhosis or WKS). Altered gene expression was found in microarray comparisons of alcoholic and control SFC samples [1]. We used Western blots and proteomic analysis to identify the proteins that also show differential expression. Tissue was obtained at autopsy under informed, written consent from uncomplicated alcoholics and age- and sex-matched controls. Alcoholics had consumed 80 g ethanol/day chronically (often, 200 g/day for 20 y). Controls either abstained or were social drinkers ( 20 g/day). All subjects had pathological confirmation of liver and brain diagnosis; none had been polydrug abusers. Samples were homogenized in water and clarified by brief centrifugation (1000g, 3 min) before storage at –80°C. For proteomics the thawed suspensions were centrifuged (15000g, 50 min) to prepare soluble fractions. Aliquots were pooled from SFC samples from the 5 chronic alcoholics and 5 matched controls used in the previous microarray study [1]. 2-Dimensional electrophoresis was performed in triplicate using 18 cm format pH 4–7 and pH 6–11 immobilized pH gradients for firstdimension isoelectric focusing. Following second-dimension SDS-PAGE the proteins were fluorescently stained and the images collected by densitometry. 182 proteins differed by 2-fold between cases and controls. 141 showed lower expression in alcoholics, 33 higher, and 8 were new or had disappeared. To date 63 proteins have been identified using MALDI-MS and MS-MS. Western blots were performed on uncentrifuged individual samples from 76 subjects (controls, uncomplicated alcoholics and cirrhotic alcoholics). A common standard was run on every gel. After transfer, immunolabeling, and densitometry, the intensities of the unknown bands were compared to those of the standards. We focused on proteins from transcripts that showed clear differences in a series of microarray studies, classified into common sets including Regulators of G-protein Signaling and Myelin-associated proteins. The preponderantly lower level of differentially expressed proteins in alcoholics parallels the microarray mRNA analysis in the same samples. We found that mRNA and protein expression do not frequently correspond; this may help identify pathogenic processes acting at the level of transcription, translation, or post-translationally.