Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding

The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120SU plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this “glycan shield” can help protect the virus from antibody-mediated neutralization. In recent years, however, it has become clear that gp120 glycosylation can also be included in the targets of recognition by some of the most potent broadly neutralizing antibodies. Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. These results should help guide the design of vaccine immunogens

HIV-1 is budded from CD4+ T lymphocytes independently of exosomes

The convergence of HIV-1 budding and exosome biogenesis at late endosomal compartments called multivesicular bodies has fueled the debate on whether HIV-1 is budded from its target cells and transmitted in the form of exosomes. The point of contention appears to primarily derive from the types of target cells in question and lack of a well-defined protocol to separate exosomes from HIV-1. In this study, we adapted and established a simplified protocol to define the relationship between HIV-1 production and exosome biogenesis. Importantly, we took advantage of the newly established protocol to unequivocally show that HIV-1 was produced from CD4+ T lymphocytes Jurkat cells independently of exosomes. Thus, this study not only presents a simplified way to obtain highly purified HIV-1 virions for identification of host proteins packaged into virions, but also provides a technical platform that can be employed to define the relationship between exosome biogenesis and budding of HIV-1 or other viruses and its contributions to viral pathogenesis

Current clinical applications of spectral tissue Doppler echocardiography (E/E' ratio) as a noninvasive surrogate for left ventricular diastolic pressures in the diagnosis of heart failure with preserved left ventricular systolic function

Congestive heart failure with preserved left ventricular systolic function has emerged as a growing epidemic medical syndrome in developed countries, which is characterized by high morbidity and mortality rates. Rapid and accurate diagnosis of this condition is essential for optimizing the therapeutic management. The diagnosis of congestive heart failure is challenging in patients presenting without obvious left ventricular systolic dysfunction and additional diagnostic information is most commonly required in this setting. Comprehensive Doppler echocardiography is the single most useful diagnostic test recommended by the ESC and ACC/AHA guidelines for assessing left ventricular ejection fraction and cardiac abnormalities in patients with suspected congestive heart failure, and non-invasively determined basal or exercise-induced pulmonary capillary hypertension is likely to become a hallmark of congestive heart failure in symptomatic patients with preserved left ventricular systolic function. The present review will focus on the current clinical applications of spectral tissue Doppler echocardiography used as a reliable noninvasive surrogate for left ventricular diastolic pressures at rest as well as during exercise in the diagnosis of heart failure with preserved left ventricular systolic function. Chronic congestive heart failure, a disease of exercise, and acute heart failure syndromes are characterized by specific pathophysiologic and diagnostic issues, and these two clinical presentations will be discussed separately

Direct and indirect interactions between ants ( Pheidole megacephala ), scales ( Coccus viridis ) and plants ( Pluchea indica )

This study investigated direct and indirect interactions between the ant, Pheidole megacephala (Fabr.), the green scale, Coccus viridis (Green), and the scale's host plant, Pluchea indica (L.). To examine the influence of ants on scales and host plants, scale population densities, scale mortality rates, and plant performance were studied on control host plants with ants and host plants from which ants had been removed. Plants with ants present had significantly greater scale population densities and scale reproductive rates than did plants without ants. Scale mortality from both parasitism and other causes was increased on plants without ants relative to plants with ants. Predator introduction experiments showed that P. megacephala removes predatory coccinellid larvae, even when they are covered with a protective coating. Host plants from which ants had been removed had significantly higher degrees of honeydew accumulation, which resulted in greater colonization by sooty mold and greater rates of leaf death and abscission. Ants also removed herbivorous lepidopteran larvae from plants. Results are discussed in terms of the potential of P. megacephala to exert direct and indirect positive effects on scale populations and an indirect positive effect on Pluchea indica .Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47790/1/442_2004_Article_BF00325261.pd

Newborn Genetic Screening for Hearing Impairment: A Preliminary Study at a Tertiary Center

Universal newborn hearing screening (UNHS) is of paramount importance for early identification and management of hearing impairment in children. However, infants with slight/mild, progressive, or late-onset hearing impairment might be missed in conventional UNHS. To investigate whether genetic screening for common deafness-associated mutations could assist in identifying these infants, 1017 consecutive newborns in a tertiary hospital were subjected to both newborn hearing screening using a two-step distortion-product otoacoustic emissions (DPOAE) screening and newborn genetic screening (NGS) for deafness. The NGS targeted 4 deafness-associated mutations commonly found in the Taiwanese population, including p.V37I (c.109G>A) and c.235delC of the GJB2 gene, c.919-2A>G of the SLC26A4 gene, and mitochondrial m.1555A>G of the 12S rRNA gene. The results of the NGS were then correlated to the results of the NHS. Of the 1017 newborns, 16 (1.6%) had unilateral DPOAE screening failure, and 22 (2.2%) had bilateral DPOAE screening failure. A total of 199 (19.6%) babies were found to have at least 1 mutated allele on the NGS for deafness, 11 (1.1%) of whom were homozygous for GJB2 p.V37I, 6 (0.6%) compound heterozygous for GJB2 p.V37I and c.235delC, and 1 (0.1%) homoplasmic for m.1555A>G, who may potentially have hearing loss. Among them, 3 babies, 5 babies, and 1 baby, respectively, passed the NHS at birth. Comprehensive audiological assessments in the 9 babies at 3 months identified 1 with slight hearing loss and 2 with mild hearing loss. NGS for common deafness-associated mutations may identify infants with slight/mild or potentially progressive hearing impairment, thus compensating for the inherent limitations of the conventional UNHS

Exosomal Shuttle RNA

Exosomes are small membrane nanovesicles of endocytic origin that can be released by many different cells to the extracellular environment. Exosomes have been found in a number of body fluids such as blood plasma, breast milk, bronchoalveolar lavage fluid and urine, indicating relevance in vivo. Exosomes have been suggested to have a number of different functions and are believed to take part in the communication between cells. Previously, exosomes were believed to consist of a lipid bilayer and proteins, but no nucleic acids. The aim of this thesis was to assess the composition and functions of mast cell exosomes, with focus on the content of nucleic acids and cell to cell communication. We utilized exosomes released from two mast cell lines as well as mouse primary bone marrow derived mast cells. Exosomes were isolated and detected as small 40-80 nm membrane vesicles, which were positive for the tetraspanins CD9, CD63 and CD81 as assessed by electron microscopy and flow cytometry. We showed for the first time that mast cell exosomes contain RNA but no DNA. The exosomal RNA differs from the donor cell RNA. Exosomes contain very little or no ribosomal RNA but a substantial amount of small RNA. We further characterized the RNA using Affymetrix DNA microarray and microRNA array analysis, which revealed that exosomes contain a selection of both microRNA and mRNA. Interestingly, a number of mRNAs were detected in the exosomes but not in their donor cells. Transfer experiments revealed that the exosomal RNA is shuttled to other mast cells and to CD34 positive progenitor cells. Exosomal RNA is functional, as shown by in vitro translation and the translation of mouse exosomal RNA to mouse protein after transfer to a human mast cell. In summary, mast cell exosomes contain mRNAs and microRNAs, which can be delivered to another cell. Exosomal RNA shuttle may be a powerful mode of communication between cells, either in the microenvironment or over a distance. We propose that this RNA be called “exosomal shuttle RNA” (esRNA)

Caracterização da normalidade do P300 em adultos jovens Standard characterization of P300 in young adults

OBJETIVO: Comparar com a literatura os valores de latência e amplitude de P300 em adultos jovens brasileiros saudáveis a fim de estabelecer um padrão normativo para futuros estudos, e verificar se os valores encontrados na amostra possuem correlação com o sexo, situação proposta na metodologia do exame e fases do ciclo menstrual. MÉTODOS: A amostra constou de 22 indivíduos saudáveis, sem queixa de desatenção, sem problemas neurológicos e auditivos, de 18 a 30 anos de idade. Todos os indivíduos foram submetidos à avaliação audiológica básica a fim de excluir aqueles com perda de audição. As respostas do P300 foram obtidas com equipamento AMPLAID MK22, de dois canais e em ambiente silencioso. RESULTADOS: Os resultados de latência encontrados na amostra, depois de comparados à literatura, enquadraram-se melhor nos intervalos de latência entre 220 e 380 ms. Somente os valores de amplitude foram influenciados pelas variáveis sexo e fase do ciclo menstrual. CONCLUSÃO: Os valores de latência da amostra não sofreram influência da variável sexo, ciclo menstrual e situação, e aproximaram-se com mais fidedignidade aos valores de intervalos de latência entre 220 e 380 ms. Contudo, os intervalos para a amplitude sofreram influência das variáveis sexo e ciclo menstrual, mas não se enquadraram adequadamente aos valores de intervalo encontrados na literatura.<br>PURPOSE: To compare the values of latency and amplitude of P300 of healthy Brazilian young adults with the data available on the literature, in order to establish a normative standard for future studies. In addition, the aim was also to verify if the values found in the sample were correlated to gender, situation proposed in the methodology of the examination, and phase of menstrual cycle. METHODS: The sample consisted of 22 healthy individuals, without any attention complaints, neurological or auditory disorders, and with ages ranging from 18 to 30 years. All subjects carried out basic audiological evaluation in order to exclude those with hearing loss. The P300 test was carried out in a quiet room, using the two channels equipment AMPLAID MK22. RESULTS: The latency values obtained in the sample, after compared with the literature reference, were in accordance with the latency intervals between 220 e 380 ms. Only amplitude values were influenced by gender and phase of the menstrual cycle. CONCLUSION: The latency values of the sample were not influenced by the variables gender, methodology of the testing situation and menstrual cycle, and were in accordance to the latency interval values between 220 and 380 ms. Amplitude values were influenced by gender and menstrual cycle, but were not in accordance to the values found in literature