Detection of HIV and HCV RNA in semen from Brazilian coinfected men using multiplex PCR before and after semen washing

INTRODUCTION: Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests. METHODS: After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen. RESULTS: In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions. CONCLUSIONS: Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim-up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals.

Carbon isotopic ratio analysis by gas chromatography/combustion/isotope ratio mass spectrometry for the detection of gamma-hydroxybutyric acid (GHB) administration to humans.

Since GHB (gamma-hydroxybutyric acid) is naturally produced in the human body, clinical and forensic toxicologists must be able to discriminate between endogenous levels and a concentration resulting from exposure. To suggest an alternative to the use of interpretative concentration cut-offs, the detection of exogenous GHB in urine specimens was investigated by means of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). GHB was isolated from urinary matrix by successive purification on Oasis MCX and Bond Elute SAX solid-phase extraction (SPE) cartridges prior to high-performance liquid chromatography (HPLC) fractioning using an Atlantis dC18 column eluted with a mixture of formic acid and methanol. Subsequent intramolecular esterification of GHB leading to the formation of gamma-butyrolactone (GBL) was carried out to avoid introduction of additional carbon atoms for carbon isotopic ratio analysis. A precision of 0.3 per thousand was determined using this IRMS method for samples at GHB concentrations of 10 mg/L. The (13)C/(12)C ratios of GHB in samples of subjects exposed to the drug ranged from -32.1 to -42.1 per thousand, whereas the results obtained for samples containing GHB of endogenous origin at concentration levels less than 10 mg/L were in the range -23.5 to -27.0 per thousand. Therefore, these preliminary results show that a possible discrimination between endogenous and exogenous GHB can be made using carbon isotopic ratio analyses.

Labile and stable fractions of soil organic matter under management systems and native cerrado

Soil organic matter can be analyzed on the basis of the different fractions. Changes in the levels of organic matter, caused by land use, can be better understood by alterations in the different compartments. The aim of this study was to evaluate the effect of different management systems on the labile and stable organic matter of a dystrophic Red Latosol (Oxisol). The following properties were determined: total organic C and total N (TOC and TN), particulate organic C and particulate N (POC and PN), organic C and N mineral-associated (MOC and NM) and particulate organic C associated with aggregate classes (POCA). Eight treatments were used: seven with soil management systems and one with native Cerrado as a reference. The experiment was designed to study the dynamics of systems of tillage and crop rotation, alternating in time and space. The experimental design was a randomized block design with three replications. The soil samples were collected from five depths: 0-5, 5-10, 10-20, 20-30 and 30-40 cm. Changes in organic C by land use occurred mainly in the fraction of particulate organic matter (> 53 mm). Proper management of grazing promoted increased levels of particulate organic matter by association with larger aggregates (2-8 mm), demonstrating the importance of the formation of this aggregate class for C protection in pasture.

Partikkelikokoanalyysimenetelmän kehittäminen

Päällystettyä paperia valmistettaessa syntyy päällystettyä hylkyä, joka kierrätetään takaisin prosessiin raaka-aineen tehokkaaksi hyödyntämiseksi. Hylyn mukana takaisin paperikoneen lyhyeen kiertoon päätyy myös pigmenttiaines levymäisinä partikkeleina. Nämä partikkelit rejektoituvat lyhyen kierron pyörrepuhdistimilla. Raaka-aineen hävikin pienentämiseksi käytetään lyhyessä kierrossa täyteaineen talteenottojärjestelmää, jonka tehtävänä on hienontaa päällystysainepartikkelit tasakokoisiksi, jotta ne voitaisiin palauttaa prosessiin. Talteenottolaitteistojen toiminnan tarkkailun kannalta on keskeistä tietää pyörrepuhdistuslaitoksen eri jakeiden partikkelikokojakaumat juuri pigmenttipartikkelien osalta. Tätä määritystä häiritsee näytteissä oleva kuitu. Tässä työssä pyrittiin löytämään partikkelikokoanalyysimenetelmä, jolla pigmenttien partikkelikokojakauma saataisiin selvitettyä kuidusta huolimatta. Aiemmin käytetty näytteen tuhkaus esikäsittelynä ennen partikkelikokoanalyysiä laserdiffraktiometrillä on osoittautunut toimimattomaksi. Kokeiden pääpaino keskittyi näytteen esikäsittelyyn fraktioinnilla ennen laserdiffraktioanalyysiä ja virtaussytometriamittauksiin. Fraktiointiin käytettiin DDJ-laitetta (dynamic drainage jar), joka oli varustettu metalliviiralla. Kumpikaan menetelmistä ei ollut täysin toimiva partikkelikokoanalyysiin, fraktioinnilla saadaan vähennettyä kuidun partikkelikokojakaumaan aiheuttamaa virhettä, mutta sen toimivuus riippuu paljolti näytteestä. Virtaussytometrialla väriainetta SYTO13 käyttämällä saadaan pigmenttipartikkelit tunnistettua ja näin rajattua kuidut pois mittauksista, mutta pigmenttiä ei saada erotettua puuperäisestä hienoaineesta, mikä vääristää mittaustulosta.

Massan fraktioiden vaikutus paperin bulkin ja sileyden väliseen yhteyteen

Diplomityön tavoitteena oli selvittää paperin valmistuksessa käytettävien mekaanisten massojen eri kuitufraktioiden vaikutusta SC-paperin ominaisuuksiin. Aikakausipaperin vaatimuksena on hyvä paperin sileys, kiilto ja bulkki. Sileys ja kiilto on helppo saavuttaa kalanteroimalla, mutta tällöin menetetään paperin bulkkia. Tässä työssä keskitytäänkin etsimään keinoja paperin bulkin säästämiseksi fraktioinnin avulla. Työn kirjallisuusosassa käydään läpi teoriaa paperin valmistuksesta ja paperin ominaisuuksista. Työn kokeellisessa osassa on esitetty tehdyt tutkimukset laboratoriomenetelmin kalanterointiolosuhteiden ja massojen eri fraktioiden vaikutuksesta paperin ominaisuuksiin. Saadut tulokset olivat samantyyppisiä mitä edellisissä tutkimuksissa samasta aiheesta oli saatu. Saatujen tulosten mukaan paperin pitkäkuitufraktion osuutta kasvattamalla bulkki ja taivutusjäykkyys paranivat, mutta toisaalta sileys ja kiilto huononivat. Paperin hienoainefraktion osuutta kasvattamalla kaikki painatusominaisuudet paranivat kuten sileys, kiilto ja valonsirontakerroin, mutta samalla huononivat paperin bulkki ja taivutusjäykkyys.

Perfil eletroforético de proteínas séricas do sangue do cordão umbilical de cães

A importância do estudo do sangue do cordão umbilical (SCU) tem sido verificada pela presença de células-tronco hematopoéticas no SCU humano. O modelo experimental em cão tem propiciado informações importantes nos transplantes em humanos. Entretanto, apesar da importância do SCU e do modelo experimental em cães, não existem estudos sobre a eletroforese de proteínas séricas do cordão umbilical canino. No presente protocolo experimental, foi feita a colheita de SCU de 20 neonatos caninos, o qual foi utilizado para o fracionamento eletroforético de suas proteínas. Os valores médios absolutos obtidos para proteínas séricas totais, albumina, alfa, beta e gamaglobulinas no sangue do cordão umbilical de cães ao nascimento foram 3,09±0,59; 1,51±0,38; 0,87±0,17; 0,68±0,12 e 0,03±0,01, respectivamente. Todos os resultados apresentaram-se abaixo dos níveis reportados para animais adultos, devido à passagem das proteínas e suas frações ocorrerem principalmente através do colostro e pela imaturidade hepática. em todos os traçados eletroforéticos do SCU de cães, foi observada uma pequena curva entre alfaglobulina 2 e betaglobulina 1, não relatada no soro de cães adultos. Portanto, neste experimento, foram observadas diferenças quantitativas no traçado eletroforético das proteínas séricas do SCU de cães, ao nascimento, diferindo-os, neste particular, de cães em diferentes fases da vida pós-colostral.

Perfil eletroforético do proteinograma sérico de eqüinos com obstrução experimental do cólon menor

Avaliaram-se as alterações do proteinograma sérico de eqüinos submetidos à isquemia e reperfusão do cólon menor por distensão intraluminal. Foram utilizados 10 animais submetidos à laparotomia pelo flanco, em posição quadrupedal, para a indução de obstrução no cólon menor durante um período de quatro horas. Cinco animais foram instrumentados, mas sem distensão (grupo controle - G1). em cinco outros animais, foi realizada isquemia mural por distensão do cólon menor via manguito inflado com 40mmHg (grupo distendido - G2). Foram colhidas amostras de sangue antes da intervenção cirúrgica (M1), com 4 horas da colocação do manguito (M2) e com 3 horas (M3) e 12 horas (M4) de reperfusão. Após centrifugação e fracionamento das amostras, as proteínas de fase aguda foram separadas por eletroforese em gel de poliacrilamida contendo SDS-PAGE, e suas concentrações determinadas por densitometria computadorizada. Foram encontradas 19 proteínas no fracionamento eletroforético, com peso molecular variando de 185.000 a 14.000 Daltons (Da). Os pesos moleculares encontrados, correspondentes às proteínas mais conhecidas, foram ceruloplasmina, 130.000 Da; proteína C-reativa, 122.000 Da; transferrina, 85.000 Da; α1-antitripsina, 61.000 Da; haptoglobina, 47.000 Da; e glicoproteína ácida, 40.000 Da. Os resultados mostram que proteínas de fase aguda se alteram após o trauma cirúrgico.

Phosphorus sources and fractions in an oxisol under no-tilled soybean

A dinâmica do P no solo pode ser modificada em sistemas com semeadura direta. Os agricultores brasileiros vêm aplicando fósforo na superfície do solo para otimizar a operação de máquinas, embora não seja uma prática recomendada pela pesquisa. Foi conduzido um experimento para estudar os efeitos da aplicação de duas fontes de fósforo na superfície do solo e no sulco de semeadura da soja. Os tratamentos consistiram da aplicação ou não de 80 kg ha-1 de P total como fosfato natural reativo e superfosfato, aplicados na superfície do solo, sobre os resíduos da cultura anterior. Na semeadura da soja, tratamentos adicionais (80 kg ha-1 de fosfato natural ou superfosfato) foram aplicados ao sulco de semeadura. O solo foi amostrado até 40 cm de profundidade, antes da semeadura da soja e após a colheita. Uma amostra controle foi tomada de área adjacente, sem cultivo. Houve aumento nos teores de P até 40 cm de profundidade, após a colheita da soja, de modo que o aumento foi observado principalmente no P ligado ao Ca e no P orgânico. Entretanto houve decréscimo nos teores de P ligado ao Fe. A adubação com ambas as fontes de fosfato levaram a um decréscimo nos teores de P ocluso na superfície do solo quando comparada com a área sem cultivo.

Métodos e estratégias em proteômica e suas aplicações na área vegetal

A implementação da espectrometria de massa (MS) para as análises de peptídeos (MS) e de aminoácidos (MS em tandem ou MS/MS) tornou possível a identificação de centenas de proteínas em experimentos únicos. Uma grande variedade de estratégias está disponível atualmente para o fracionamento e a purificação de amostras, a identificação de proteínas, a quantificação, a análise de modificações pós-traducionais (MPT's) e os estudos de interação. Dessa forma, a proteômica abre novas perspectivas na biologia de plantas com ênfase nos estudos de variabilidade genética, estresses fisiológicos e desenvolvimento de plantas.

DSC studies on protein isolate of guava seeds Psidium guajava

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Gas chromatographic analysis of indole alkaloids from Tabernaemontana hilariana

A fast and efficient procedure was elaborated to identify the alkaloid constituents from Tabernaemontana hilariana (Apocynaceae). The strategy based on fractioning of the crude alkaloid fraction in small silica cartridges followed by thin-layer chromatography (TLC), capillary gas chromatography-flame ionization detection as well as high-resolution gas chromatography-mass spectrometry afforded voacangine, coronaridine, ibogamine, voacangine pseudoindoxyl, voacangine hydroxyindolenine, 3-hydroxycoronaridine and 3-(2-oxopropyl)coronaridine. (C) 1997 Elsevier B.V. B.V.

Soil microbial biomass and organic matter fractions during transition from conventional to organic farming systems

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Prescriptionless light-cone integrals

Perturbative quantum gauge field theory as seen within the perspective of physical gauge choices such as the light-cone gauge entails the emergence of troublesome poles of the type (k · n)-α in the Feynman integrals. These come from the boson field propagator, where α = 1, 2, ⋯ and nμ is the external arbitrary four-vector that defines the gauge proper. This becomes an additional hurdle in the computation of Feynman diagrams, since any graph containing internal boson lines will inevitably produce integrands with denominators bearing the characteristic gauge-fixing factor. How one deals with them has been the subject of research over decades, and several prescriptions have been suggested and tried in the course of time, with failures and successes. However, a more recent development at this fronteer which applies the negative dimensional technique to compute light-cone Feynman integrals shows that we can altogether dispense with prescriptions to perform the calculations. An additional bonus comes to us attached to this new technique, in that not only it renders the light-cone prescriptionless but, by the very nature of it, it can also dispense with decomposition formulas or partial fractioning tricks used in the standard approach to separate pole products of the type (k · n)-α[(k - p) · n]-β (β = 1, 2, ⋯). In this work we demonstrate how all this can be done.

Attractiveness of citrus pulp and orange albedo extracts to Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The present study aimed to compare the attractiveness of industrial citrus pulp with the handmade orange albedo to the workers of Atta sexdens rubropilosa. For this, filter paper fragments were impregnated with organic extracts obtained through chemical extraction and sequential fractioning with hexane and dichloromethane and offered to different field nests. It was verified that the industrial citrus pulp extract is as good as the handmade orange albedo extract. This preference is discussed keeping in mind the chemical, behavioral and nutritional factors.

Os granitóides brasilianos da faixa de dobramentos paraguai, MS e MT

The Brazilian Granitic Province from southeastern Mato Grosso do Sul and Mato Grosso region, central western Brazil, can be divided into two major groups and/or magmatic events related to the evolution of the Paraguay Fold Belt. The southern portion crops out in Mato Grosso do Sul State and is constituted by the Taboco, Rio Negro, Coxim and Sonora massifs forming NE-SW oriented, elongated small intrusions. The north portion crops out in Mato Grosso State and is constituted by the São Vicente, Araguaiana and Lajinha batholiths. Lithogeochemical aspects of the northern granites point to Type-I granites ranging from K calc-alkaline to high-K, peraluminous to metaluminous in composition, generated in an environment of continental collision and/or post- collision decompression. The southern granites are Type-I, from K calc-alkaline to high-K, peraluminous to subordinate metalummous, in a syn-collision continental arc environment with the exception of some pre-collisional facies from the Rio Negro Massif. The southern granites have less SiO 2 and K 2O, and are less differentiated and evolved than granites from the northern region. The four southern granites can be grouped into two subordinate sets with the degree of differentiation increasing from South (Taboco and Rio Negro) to North (Coxim and Sonora). The granitic rocks are characterized by a magmatism generated by melting of material from the lower crust which suggests that in this province the formation from non-cogenetic magmas with diversified compositions and distinct degrees of fractioning reaching more steady consolidated environments at the end of the collisional event in the southeastern Amazonian Craton.