Determinação simultânea de ácido kójico e hidroquinona por espectrofotometria visível e calibração multivariada

A multivariate spectrophotometric method was developed for analysis of kojic acid/hydroquinone associations in skin whitening cosmetics. The method is based on the reaction between kojic acid and Fe3+ and on the reduction of Fe3+ by hydroquinone and further complexation of Fe2+ with 1,10-phenanthroline. The multivariate model was developed by Partial Least Squares Regression (PLSR), using 25 synthetic mixtures and mean-centered spectral data (350-380 nm). The use of 3 (kojic acid) and 2 (hydroquinone) latent variables permits the observation of mean errors of about 5% in the external validation phase.

Monitorando atividades enzimáticas com sondas fluorogênicas

Latent fluorogenic probes are essential tools for molecular and chemical biology, providing valuable information about enzymatic activity and occurrence. In this review, a brief outline of fluorophores and latent fluorogenic probes is given. Furthermore, advances and challenges in the development of fluorogenic chemical probes to visualize enzymatic activities (hydrolases and oxidoreductases) of biotechnological and biomedical interest are highlighted, including some methodologies for intracellular imaging.

Garlic viral complex: identification of Potyviruses and Carlavirus in Central Brazil

Garlic viruses often occur in complex infections in nature. In this study, a garlic virus complex, collected in fields in Brazil, was purified. RT-PCR was performed using specific primers designed from the consensus regions of the coat protein genes of Onion yellow dwarf virus, a garlic strain (OYDV-G) and Leek yellow stripe virus (LYSV). cDNA of Garlic common latent virus (GCLV) was synthesized using oligo-dT and random primers. By these procedures individual garlic virus genomes were isolated and sequenced. The nucleotide sequence analysis associated with serological data reveals the presence of two Potyvirus OYDV-G and LYSV, and GCLV, a Carlavirus, simultaneously infecting garlic plants. Deduced amino acid sequences of the Brazilian isolates were compared with related viruses reported in different geographical regions of the world. The analysis showed closed relations considering the Brazilian isolates of OYDV-G and GCLV, and large divergence considering LYSV isolate. The detection of these virus species was confirmed by specific reactions observed when coat protein genes of the Brazilian isolates were used as probes in dot-blot and Southern blot hybridization assays. In field natural viral re-infection of virus-free garlic was evaluated.

Sequence analysis of the capsid protein gene of an isolate of Apple stem grooving virus, and its survey in Southern Brazil

Apple stem grooving virus (ASGV) is one of the most important viruses infecting fruit trees. This study aimed at the molecular characterization of ASGV infecting apple (Malus domestica) plants in Santa Catarina (SC). RNA extracted from plants infected with isolate UV01 was used as a template for RT-PCR using specific primers. An amplified DNA fragment of 755 bp was sequenced. The coat protein gene of ASGV isolate UV01 contains 714 nucleotides, coding for a protein of 237 amino acids with a predicted Mr of approximately 27 kDa. The nucleotide and the deduced amino acid sequences of the coat protein gene showed identities of 90.9% and 97.9%, respectively, with a Japanese isolate of ASGV. Very high amino acid homologies (98.7%) were also found with Citrus tatter leaf capillovirus (CTLV), a very close relative of ASGV. These results indicate low coat protein gene variability among Capillovirus isolates from distinct regions. In a restricted survey, mother stocks in orchards and plants introduced into the country for large scale fruit production were indexed and shown to be infected by ASGV (20%), usually in a complex with other (latent) apple viruses (80%).

Detecção de um isolado de Grapevine virus A e caracterização do gene da proteína capsidial

O Grapevine virus A (GVA) está associado à "Acanaladura do lenho de Kober", uma doença do complexo rugoso da videira (Vitis spp.). Neste trabalho, um isolado brasileiro de GVA (GVA-RS) foi caracterizado biologicamente por transmissão mecânica para cinco hospedeiras herbáceas e por enxertia na videira indicadora cv. Kober 5BB, e também por sorologia. O RNA total foi extraído de videira infetada cv. Pirovano 65. Para a RT-PCR, dois pares de oligonucleotídeos foram utilizados. Dois fragmentos de DNA, 430 e 451 pb, apresentando sobreposição parcial de nucleotídeos, foram amplificados por PCR. A seqüência do gene da proteína capsidial do GVA-RS com 597 nucleotídeos e 198 aminoácidos deduzidos, com massa molecular calculada de 21,6 kDa, foi alinhada a outros isolados virais. As seqüências de nucleotídeos e aminoácidos deduzidos do GVA-RS apresentaram maior identidade, 91,4% e 95,4%, respectivamente, com um isolado italiano. O GVA-RS apresentou expressiva divergência dos Vitivirus Heracleum latent virus (HLV), Grapevine virus B (GVB) e Grapevine virus D (GVD), com identidade de nucleotídeos variando de 76% a 83,1%.

Polyclonal antibodies to the coat protein of Apple stem grooving virus expressed in Escherichia coli: production and use in immunodiagnosis

The coat protein gene of Apple stem grooving virus (ASGV) was amplified by RT-PCR, cloned, sequenced and subcloned in the expression vector pMal-c2. This plasmid was used to transform Escherichia coli BL21c+ competent cells. The ASGV coat protein (cp) was expressed as a fusion protein containing a fragment of E. coli maltose binding protein (MBP). Bacterial cells were disrupted by sonication and the ASGVcp/MBP fusion protein was purified by amylose resin affinity chromatography. Polyclonal antibodies from rabbits immunized with the fusion protein gave specific reactions to ASGV from infected apple (Malus domestica) cv. Fuji Irradiada and Chenopodium quinoa at dilutions of up to 1:1,000 and 1:2,000, respectively, in plate trapped ELISA. The ASGVcp/MBP fusion protein reacted to a commercial antiserum against ASGV in immunoblotting assay. The IgG against ASGVcp/MBP performed favorably in specificity and sensitivity to the virus. This method represents an additional tool for the efficient ASGV-indexing of apple propagative and mother stock materials, and for use in support of biological and molecular techniques.

Epidemiological analysis of clones and cultivars of potato in soil naturally infested with Ralstonia solanacearum biovar 2

The objectives of this study were to evaluate the progress of Ralstonia solanacearum bacterial potato wilt biovar 2 (race 3) in 14 potato (Solanum tuberosum L.) cultivars or clones, the resistance of potato clone MB 03 (selected in Brasília, Brazil) to race 1 of R. solanacearum, and the occurrence of the pathogen in tubers harvested from asymptomatic potato plants. During the spring (September to the end of November in the southern hemisphere) of 1999 and 2000, 14 cultivars or clones were grown in a field naturally infested with R. solanacearum biovar 2, in Caxias do Sul, RS. The number of wilted potato plants was recorded each week and a disease progress curve plotted, the resistance of the potato genotypes to bacterial wilt being evaluated by determining the area under the curve. Various models were evaluated to fit the curves, with the logistic model being the best fit. At the end of each growing season tubers produced by asymptomatic plants were harvested and stored until budding and then tested for the presence of R. solanacearum. Cultivar Cruza 148 and clone MB 03 were the most resistant but both showed tubers with latent infections. The epidemiological implications of the incidence of R. solanacearum biovar 2 (race 3) in potato crops, as well as the resistance of certain genotypes that may harbor latent infections, are important aspects to be considered in the integrated management of bacterial wilt.

Effect of temperature, wetness duration and cultivar on the development of anthracnose in guava fruits

The development of a large number of postharvest diseases is closely associated with fruit ripeness. Environmental conditions may affect both the pathogen development and the fruit ripening rate. The aim of this study was to determine the most favorable temperature and wetness duration to the development of anthracnose in guava fruits. Cultivars 'Kumagai' (white pulp) and 'Pedro Sato' (red pulp) were inoculated with a conidial suspension of Colletotrichum gloeosporioides and C. acutatum and incubated at constant temperature ranging from 10 to 35ºC and wetness duration of 6 and 24 hours. Disease severity and incidence were evaluated at every two days during 12 days. No infection occurred at 10 and 35ºC, regardless of the wetness duration. The optimum conditions for fruit infection were 26 and 27ºC for 'Kumagai' and 25 and 26ºC for 'Pedro Sato', adopting 24 hours of wetness. In general, the disease development in 'Kumagai' cultivar was more affected by the wetness period, compared to 'Pedro Sato'. Disease severity for 'Kumagai' fruits was maximal between 25 and 30ºC , depending on the Colletotrichum species. Regarding 'Pedro Sato', the mean diameter of lesions was greater in fruits stored at 20, 25 and 30ºC , compared to 'Kumagai' cultivar, depending on the wetness period and the species. The incubation period (between 6 and 7 days) and the latent period (between 8 and 10 days) were minimal at 30ºC. The data generated in this study will be useful either for the development of a disease warning system or for the increase in the shelf life of guavas in the postharvest.

Estimation of the potential evapotranspiration by a simplified penman method

The numerous methods for calculating the potential or reference evapotranspiration (ETo or ETP) almost always do for a 24-hour period, including values of climatic parameters throughout the nocturnal period (daily averages). These results have a nil effect on transpiration, constituting the main evaporative demand process in cases of localized irrigation. The aim of the current manuscript was to come up with a model rather simplified for the calculation of diurnal daily ETo. It deals with an alternative approach based on the theoretical background of the Penman method without having to consider values of aerodynamic conductance of latent and sensible heat fluxes, as well as data of wind speed and relative humidity of the air. The comparison between the diurnal values of ETo measured in weighing lysimeters with elevated precision and estimated by either the Penman-Monteith method or the Simplified-Penman approach in study also points out a fairly consistent agreement among the potential demand calculation criteria. The Simplified-Penman approach was a feasible alternative to estimate ETo under the local meteorological conditions of two field trials. With the availability of the input data required, such a method could be employed in other climatic regions for scheduling irrigation.

Mathematical simulation parameters for drying of cassava starch pellets

Based on experimental tests, it was obtained the equations for drying, equilibrium moisture content, latent heat of vaporization of water contained in the product and the equation of specific heat of cassava starch pellets, essential parameters for realizing modeling and mathematical simulation of mechanical drying of cassava starch for a new technique proposed, consisting of preformed by pelleting and subsequent artificial drying of starch pellets. Drying tests were conducted in an experimental chamber by varying the air temperature, relative humidity, air velocity and product load. The specific heat of starch was determined by differential scanning calorimetry. The generated equations were validated through regression analysis, finding an appropriate correlation of the data, which indicates that by using these equations, can accurately model and simulate the drying process of cassava starch pellets.

Energy balance in the poultry-shed system and its influence on broiler performance

ABSTRACTThe objective of this study was to determine the energy balance of the poultry-shed system and its effect on broiler performance during the production cycle. The experimental design was completely random with sub-divided blocks. The blocks were composed of five different types of sheds and the sub-blocks of the evaluation times (00:00 h to 23:00 h), allowing an analysis of variance and a comparison between means with the Tukey test. There were no significant differences between the mean values of the exchanges of sensible, latent and total heat between the poultry sheds but the differences for the evaluation times were significant (P<0.05). There was no significant difference between sheds 1 and 4 for broiler productive performance regarding weight gain, feed consumption and feed conversion. Bird performance was significant (P<0.05) for the remaining poultry sheds. The productive indexes remained below the ranges considered ideal for broilers and values in the final weeks were characterized by the poor installation efficiency in controlling temperature variations and, consequently, the energy balance in the system, which adversely affected bird productive performance.

Hérnia diafragmática traumática crônica e fístula gastropleurocutânea

Traumatic diaphragmatic hernia is defined as a laceration of the diaphragm with an abdominal viscera herniation into the thorax. It is usually asymptomatic, with the exception of the cases with obstruction, strangulation, necrosis or perforation of the herniaded viscera. It is classified as acute, latent or chronic, in accordance with the evolutive period. At the latent phase, symptoms are indefinite and the radiological signals, which are suggestive of thoracic affections, are frequent and can induce a diagnosis error, leading to inadequate treatment.This article presents a case of chronic traumatic diaphragmatic hernia which was complicated by a gastricpleuralcutaneous fistula, due to an inadequate thoracic drainage. Considering that this is a chronic affection with an unquestionable surgical indication, due to the complications risk, it is essential to have a detailed diagnostic investigation, which aims at both avoiding an intempestive or inadequate therapeutics behaviour and reducing the affection morbimortality. Recently, the videolaparoscopic approach has proved to be more precise when compared to the other diagnostic methods, by direct visualization of the diaphragmatic laceration, allowing its correction by an immediate suture.

Hérnia diafragmática traumática

Traumatic diaphragmatic hernia is an uncommon but important problem in the patient with multiple injuries. Since diaphragmatic injuries are difficult to diagnose, those that are missed may present with latent symptoms of bowel obstruction and strangulation. The same may occur in the patients with stab wounds to the lower chest. Traumatic diaphragmatic hernia should be suspected on the basis of an abnormal chest radiograph in the trauma victim with multiple injuries. This article discuss about history, epidemiology, clinical signs and symptoms, diagnostic modalities, treatment and complications.

A Brazilian glycoprotein E-negative bovine herpesvirus type 1.2a (BHV-1.2a) mutant is attenuated for cattle and induces protection against wild-type virus challenge

The authors previously reported the construction of a glycoprotein E-deleted (gE-) mutant of bovine herpesvirus type 1.2a (BHV-1.2a). This mutant, 265gE-, was designed as a vaccinal strain for differential vaccines, allowing the distinction between vaccinated and naturally infected cattle. In order to determine the safety and efficacy of this candidate vaccine virus, a group of calves was inoculated with 265gE-. The virus was detected in secretions of inoculated calves to lower titres and for a shorter period than the parental virus inoculated in control calves. Twenty one days after inoculation, the calves were challenged with the wild type parental virus. Only mild signs of infection were detected on vaccinated calves, whereas non-vaccinated controls displayed intense rhinotracheitis and shed virus for longer and to higher titres than vaccinated calves. Six months after vaccination, both vaccinated and control groups were subjected to reactivation of potentially latent virus. The mutant 265gE- could not be reactivated from vaccinated calves. The clinical signs observed, following the reactivation of the parental virus, were again much milder on vaccinated than on non-vaccinated calves. Moreover, parental virus shedding was considerably reduced on vaccinated calves at reactivation. In view of its attenuation, immunogenicity and protective effect upon challenge and reactivation with a virulent BHV-1, the mutant 265gE- was shown to be suitable for use as a BHV-1 differential vaccine virus.

Bovine herpesvirus type 5 (BHV-5) in a calf with rabies

The brain of an one year old male calf which died with signs of neurological disease was submitted to the laboratory for rabies diagnosis. Microscopical findings included moderate mielitis, mild meningoencephalitis with perivascular cell cuffing and Negri inclusion bodies in Purkinje cells of the cerebellum. Rabies virus infection was further confirmed by the direct fluorescent antibody test as well as by mouse inoculation. In addition, a herpesvirus was isolated from brain tissues. The isolate was antigenic and genetically characterized as bovine herpesvirus type 5 (BHV-5). It was not possible to determine whether BHV-5 played an active role in the outcome of the infection, since, the virus might have been present in a latent form in neural tissues. This is the first report of a mixed rabies/ BHV-5 infection in calves.