Candida albicans PROTEIN PROFILE CHANGES IN RESPONSE TO THE BUTANOLIC EXTRACT OF Sapindus saponariaL.

Candida albicans is an opportunistic human pathogen that is capable of causing superficial and systemic infections in immunocompromised patients. Extracts of Sapindus saponaria have been used as antimicrobial agents against various organisms. In the present study, we used a combination of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to identify the changes in protein abundance of C. albicans after exposure to the minimal inhibitory concentration (MIC) and sub-minimal inhibitory concentration (sub-MIC) of the butanolic extract (BUTE) of S. saponaria and also to fluconazole. A total of six different proteins with greater than 1.5 fold induction or repression relative to the untreated control cells were identified among the three treatments. In general, proteins/enzymes involved with the glycolysis (GPM1, ENO1, FBA1), amino acid metabolism (ILV5, PDC11) and protein synthesis (ASC1) pathways were detected. In conclusion, our findings reveal antifungal-induced changes in protein abundance of C. albicans. By using the previously identified components of the BUTE of S. saponaria(e.g., saponins and sesquiterpene oligoglycosides), it will be possible to compare the behavior of compounds with unknown mechanisms of action, and this knowledge will help to focus the subsequent biochemical work aimed at defining the effects of these compounds.

Critical steps in the early evolution of the isocortex: Insights from developmental biology

This article proposes a comprehensive view of the origin of the mammalian brain. We discuss i) from which region in the brain of a reptilian-like ancestor did the isocortex originate, and ii) the origin of the multilayered structure of the isocortex from a simple-layered structure like that observed in the cortex of present-day reptiles. Regarding question i there have been two alternative hypotheses, one suggesting that most or all the isocortex originated from the dorsal pallium, and the other suggesting that part of the isocortex originated from a ventral pallial component. The latter implies that a massive tangential migration of cells from the ventral pallium to the dorsal pallium takes place in isocortical development, something that has not been shown. Question ii refers to the origin of the six-layered isocortex from a primitive three-layered cortex. It is argued that the superficial isocortical layers can be considered to be an evolutionary acquisition of the mammalian brain, since no equivalent structures can be found in the reptilian brain. Furthermore, a characteristic of the isocortex is that it develops according to an inside-out neurogenetic gradient, in which late-produced cells migrate past layers of early-produced cells. It is proposed that the inside-out neurogenetic gradient was partly achieved by the activation of a signaling pathway associated with the Cdk5 kinase and its activator p35, while an extracellular protein called reelin (secreted in the marginal zone during development) may have prevented migrating cells from penetrating into the developing marginal zone (future layer I).

The calcium-dependent protease of Loxosceles gaucho venom acts preferentially upon red cell band 3 transmembrane protein

Eighty micrograms red blood cell (RBC) ghosts from patients who had previously exhibited the cutaneous form of loxoscelism (presenting localized dermonecrosis) and the viscerocutaneous form of loxoscelism (presenting dermonecrosis, hemoglobinuria, hematuria, and jaundice) and from controls were incubated with 2.5 µg crude Loxosceles gaucho venom in 5 mM phosphate buffer, pH 7.4, at 37ºC. Among all membrane proteins, quantitative proteolysis of the important integral transmembrane protein 3 increased with venom dose and with incubation time from 30 to 120 min, as demonstrated by gel densitometry. Similar quantitative data were obtained for RBC ghosts from patients and from control subjects, a fact that argues against the possibility of genetic factors favoring the hemolytic viscerocutaneous form. These data suggest that the clinical forms may be different types of the same disease, with the viscerocutaneous form being the result of large amounts of intravascularly injected venom and the superficial form being the result of in situ venom action. Since protein 3 is a housekeeping integral membrane protein, whose genetic deficiency leads to hemolytic anemia, it is reasonable to relate it to the hemolysis which occurs in the viscerocutaneous form of loxoscelism. The venom protease responsible for the process was not inhibited after 120-min incubation by 0.2 mM paramethylsulfonyl fluoride or by 0.2 mM N-ethylmaleimide but was inhibited by 25 mM ethylenediaminetetraacetic acid (a calcium-chelating agent) in 5 mM phosphate buffer at pH 7.4, which suggests that the enzyme is a calcium-dependent metalloprotease.

NMDA receptor blockade alters the intracellular distribution of neuronal nitric oxide synthase in the superficial layers of the rat superior colliculus

Nitric oxide (NO) is a molecular messenger involved in several events of synaptic plasticity in the central nervous system. Ca2+ influx through the N-methyl-D-aspartate receptor (NMDAR) triggers the synthesis of NO by activating the enzyme neuronal nitric oxide synthase (nNOS) in postsynaptic densities. Therefore, NMDAR and nNOS are part of the intricate scenario of postsynaptic densities. In the present study, we hypothesized that the intracellular distribution of nNOS in the neurons of superior colliculus (SC) superficial layers is an NMDAR activity-dependent process. We used osmotic minipumps to promote chronic blockade of the receptors with the pharmacological agent MK-801 in the SC of 7 adult rats. The effective blockade of NMDAR was assessed by changes in the protein level of the immediate early gene NGFI-A, which is a well-known NMDAR activity-dependent expressing transcription factor. Upon chronic infusion of MK-801, a decrease of 47% in the number of cells expressing NGFI-A was observed in the SC of treated animals. Additionally, the filled dendritic extent by the histochemical product of nicotinamide adenine di-nucleotide phosphate diaphorase was reduced by 45% when compared to the contralateral SC of the same animals and by 64% when compared to the SC of control animals. We conclude that the proper intracellular localization of nNOS in the retinorecipient layers of SC depends on NMDAR activation. These results are consistent with the view that the participation of NO in the physiological and plastic events of the central nervous system might be closely related to an NMDAR activity-dependent function.

Rat Dlx5 is expressed in the subventricular zone and promotes neuronal differentiation

The molecular mechanisms and potential clinical applications of neural precursor cells have recently been the subject of intensive study. Dlx5, a homeobox transcription factor related to the distal-less gene in Drosophila, was shown to play an important role during forebrain development. The subventricular zone (SVZ) in the adult brain harbors the largest abundance of neural precursors. The anterior SVZ (SVZa) contains the most representative neural precursors in the SVZ. Further research is necessary to elucidate how Dlx5-related genes regulate the differentiation of SVZa neural precursors. Here, we employed immunohistochemistry and molecular biology techniques to study the expression of Dlx5 and related homeobox genes Er81 and Islet1 in neonatal rat brain and in in vitro cultured SVZa neural precursors. Our results show that Dlx5 and Er81 are also highly expressed in the SVZa, rostral migratory stream, and olfactory bulb. Islet1 is only expressed in the striatum. In cultured SVZa neural precursors, Dlx5 mRNA expression gradually decreased with subsequent cell passages and was completely lost by passage four. We also transfected a Dlx5 recombinant plasmid and found that Dlx5 overexpression promoted neuronal differentiation of in vitro cultured SVZa neural precursors. Taken together, our data suggest that Dlx5 plays an important role during neuronal differentiation.

Functional extruded snacks with lycopene and soy protein

In this work, the effects of thermoplastic extrusion process parameters (raw material moisture content and temperature) and the addition of functional ingredients (lycopene and soy protein) on quality characteristics of a base-formulation for extruded corn snacks were studied, with the objective of developing an easy-to-eat functional product. A single-screw Labor PQ 30 model Inbramaq extruder was used for extrusion and a central composite rotational design (CCRD) was followed. The independent variables were: i) percentage of soy protein isolate (0-30%); ii) percentage of lycopene preparation (0-0.1%); iii) raw material moisture content (20-30%); and iv) 5th zone temperature (100-150 °C). The expansion index reached maximum values with the lowest raw material moisture content (20%) and intermediate temperatures (approximately 125 °C). Instrumental hardness was higher with high moisture and low temperature; however, increasing the percentage of soy protein was beneficial for the texture of the product, reducing hardness. The red color intensity increased with the increase in lycopene content and moisture, and with the reduction of temperature. Sensory acceptance tests were carried out for two products, with maximum percentages of the functional ingredients, 20% moisture and temperatures of 125 and 137 °C, with greater acceptance for the product extruded at 125 °C.

Germination, initial growth and cotyledon protein content of bean cultivars under salinity stress

Excess salts in the root zone inhibit water uptake by plants, affect nutrient uptake and may result in toxicities due to individual salts in the soil solution. Excess exchangeable sodium in the soil may destroy the soil structure to a point where water penetration and root aeration become impossible. Sodium is also toxic to many plants. Beans (Phaseolus vulgaris L.) are consumed as protein source in northeastern Brazil, although little is known about common bean cultivar tolerance to salinity. The germination of bean cultivars under salt stress was studied. The cultivars 'Carioca' and 'Mulatinho' were submitted to germination test in a germinator at 25ºC, at the Seed Analysis Laboratory of the Brazilian Agricultural Research Corporation unit in the Semi- Arid region (Embrapa Semi Árido), Petrolina, Pernambuco State. These seeds were germinated on "germitest" papers imbibed in distilled water or in 10, 50, 100 e 200 mol.m-3sodium chloride (NaCl) solutions. At the first and second counts of the germination test, normal seedlings were counted, measured, weighed and dried, supplying data for vigor, total germination, fresh matter weight and dry matter weight and seedlings length. Total protein was quantified in cotyledons at 3, 6 and 9 days after sowing. The results indicated that the NaCl content influenced seed germination and concentrations above 50 mol.m-3 decreased germination and seedling growth.

Histologia da camada superficial da lâmina própria da prega vocal ao se aplicar retalho pediculado de mucosa: estudo experimental em cães

Os resultados conseguidos até hoje para a correção de sulcos vocais e lesões cicatriciais não são universalmente aceitos. A Técnica do Retalho Pediculado de Mucosa de Prega Vocal consiste na colocação de um retalho de mucosa de prega vocal com pedículo anterior na camada superficial da lâmina própria, abaixo da borda livre. OBJETIVO: Descrever os achados histológicos pós-operatórios ocorridos na camada superficial da lâmina própria de cães ao se aplicar a técnica em questão, tomando-se como parâmetro a variação dos colágenos total, tipo I, tipo III e número de núcleos celulares. FORMA DE ESTUDO: experimental. MATERIAL E MÉTODO: Foram utilizados 15 cães. Numa das pregas foi realizada a intervenção e a contralateral foi deixada como controle. Cada grupo de três cães foi sacrificado em 10, 30, 90, 180 e 360 dias após a cirurgia. As colorações utilizadas foram: H.E. e Syrius Red. RESULTADOS: Os níveis de colágeno total e tipo I apresentaram uma tendência a aumento nos grupos de intervenção nos 90º e 180º dias de pós-operatório, contudo só houve significância estatística no 180º dia (p<0,05). A área do colágeno tipo III alcançou níveis inferiores ao do grupo controle no 180º dia (p<0,05). O número de núcleos atingiu maiores níveis no grupo teste no 10º dia de pós-operatório, seguido de decréscimo após o 30º dia. DISCUSSÃO: Os resultados encontrados quanto ao colágeno total, tipo I e tipo III e número de núcleos sugerem semelhanças ao processo cicatricial pós-operatório em laringe encontrado em outros estudos experimentais. Os presentes achados deverão ser complementados por experimentos.

Comparação da anestesia geral e bloqueio do plexo cervical superficial em tireoidectomias parciais

Tireoidectomia sob efeito de bloqueio do plexo cervical superficial (BPCS) tem sofrido resistência. OBJETIVO: Comparar variáveis cirúrgicas e anestésicas, custos do tratamento e grau de satisfação de pacientes submetidos à hemitireoidectomia sob efeito de anestesia geral e BPCS. CASUÍSTICA E MÉTODOS: Foram 21 pacientes submetidos à anestesia geral (AG) e outro tanto ao BPCS. Após sedação, no grupo com BPCS, usou-se marcaína com vasoconstritor, e quando necessário, lidocaína a 2% com vasoconstritor. Sedação intra-operatória com diazepam endovenoso e metoprolol para controle da PA e FC eram administradas quando necessário. Usou-se anestesia geral (AG) segundo padronização do serviço. RESULTADOS: Foram significantes (p<0,05, teste t de Student) para o tempo de cirurgia (ag111,4:bpcs125,5 min), tempo de anestesia (ag154,1:bpcs488,6 min), tempo de permanência na sala cirúrgica (ag15:bpcs1 min), custos do tratamento (ag203,2:bpcs87,4 R$), presença de bradicardia (ag0:bpcs23,8%) e lesão laringotraqueal (ag51:bpcs0%). Como resultados não significativos tiveram: tempo de internação (ag17,3:bpcs15,1 hora); volume de sangramento (ag41,9:bpcs47,6 gr), tamanho da peça operatória (ag52,1:bpcs93,69 cm3) e grau de satisfação dos pacientes (ag3,8:bpcs3,9). CONCLUSÃO: Embora com incidência maior de bradicardia (23,8%), o bloqueio permitiu ressecar tumorações de até 348 cm3 com menor custo e sem apresentar lesões laringotraqueais, presentes em 51% dos pacientes submetidos à AG.

Preserved xenogenic amniotic membrane as a patch on the repair of superficial corneal ulcers in rabbits

The aim of this study was to evaluate the effects of canine amniotic membrane, previously preserved in glycerin, used as a patch on the repair of experimentally-made superficial corneal ulcers and to compare corneal epithelization between the treated and non-treated groups. Xenogeneic amniotic membranes were collected aseptically and preserved in 99% glycerin at room temperature. Each animal was anesthetized and submitted to superficial corneal keratectomy of the left eye. The treated group received a fragment of canine amniotic membrane as a patch, while the control group had no treatment. The treated group showed blepharospasm, ocular discharge and conjunctival congestion. The membrane accelerated corneal repair in the beginning of the process, however, it delayed its conclusion (p<0.05). Treated eyes showed greater vessel formation and decreased corneal transparency (p<0.05). The stroma of the control group was thicker than that of the treated group (p<0.05). We suggest that amniotic membrane used in this manner can be applied as a therapy for superficial corneal ulcers in the beginning phases of the repair process.

Sugarcane productivity correlated with physical-chemical attributes to create soil management zone

The socioeconomic importance of sugar cane in Brazil is unquestionable because it is the raw material for the production of ethanol and sugar. The accurate spatial intervention in the management of the crop, resulting zones of soil management, increases productivity as well as its agricultural yields. The spatial and Person's correlations between sugarcane attributes and physico-chemical attributes of a Typic Tropustalf were studied in the growing season of 2009, in Suzanápolis, State of São Paulo, Brazil (20°28'10'' S lat.; 50°49'20'' W long.), in order to obtain the one that best correlates with agricultural productivity. Thus, the geostatistical grid with 120 sampling points was installed to soil and data collection in a plot of 14.6 ha with second crop sugarcane. Due to their substantial and excellent linear and spatial correlations with the productivity of the sugarcane, the population of plants and the organic matter content of the soil, by evidencing substantial correlations, linear and spatial, with the productivity of sugarcane, were indicators of management zones strongly attached to such productivity.

Vector competence experiments with Rocio virus and three mosquito species from the epidemic zone in Brazil

First-generation progeny of field-collected Psorophora ferox, Aedes scapularis, and Aedes serratus from the Rocio encephalitis epidemic zone in S.Paulo State, Brazil, were tested for vector competency in the laboratory. Psorophora ferox and Ae. scapularis are susceptible to per os infection with Rocio virus and can transmit the virus by bite following a suitable incubation period. Oral ID50S for the two species (10(4.1) and 10(4.3) Vero cell plaque forming units, respectively) did not differ significantly. Infection rates in Ae. serratus never exceeded 36%, and, consequently, an ID50 could not be calculated for this species. It is unlikely that Ae. serratus is an epidemiologically important vector of Rocio virus. The utility of an in vitro feeding technique for demonstrating virus transmission by infected mosquitoes and difficulties encountered in working with uncolonized progeny of field-collected mosquitoes are discussed.

Finding Aedes aegypti in a natural breeding site in an urban zone, Sao Paulo, Southeastern Brazil

ABSTRACT This is the description of how nine Aedes aegypti larvae were found in a natural breeding site in the Pinheiros neighborhood, city of Sao Paulo, SP, Southeastern Brazil. The record was conducted in December 2014, during an entomological surveillance program of dengue virus vectors, with an active search of potential breeding sites, either artificial or natural. Finding Ae. aegypti larvae in a tree hole shows this species’ ability to use both artificial and natural environments as breeding sites and habitats, which points towards the importance of maintaining continuous surveillance on this mosquito in all kinds of water-holding containers.

Immunization aganist hepatitis B in children from endemic zone: evaluation of the antibody response against the DNA recombinant vaccine (Engerix B-20 mcg)

A previous seroepidemiological study in the rural zone of Vargem Alta (ES) SouthEast of Brazil, showed a prevalence of up to 9% of hepatitis B surface antigen (HBsAg) in some areas. One hundred susceptible children aging 1 to 5 years old were selected and immunized with a recombinant DNA hepatitis B vaccine (Smith-Kline 20 mcg) using the 0-1-6 months vaccination schedule. Blood samples were collected at the time of the first vaccine dose (month 0) in order to confirm susceptible individuals and 1,3,6 and 8 months after the first dose , to evaluate the antibody response. Our results showed that two and five months after the second dose, 79% and 88% of children seroconverted respectively, reaching 97% after the third dose. The levels of anti-HBs were calculated in milli International Units/ml (mIU/ml) and demonstrated the markedly increase of protective levels of antibodies after the third dose. These data showed a good immunogenicity of the DNA recombinant hepatitis B vaccine when administered in children of endemic areas.