Outbreaks of vesicular disease caused by Vaccinia virus in dairy cattle from Goiás State, Brazil (2010-2012)

Cases of vesicular and exanthematic disease by Vaccinia virus (VACV) have been reported in dairy herds of several Brazilian regions, occasionally also affecting humans. The present article describes eight outbreaks of vesicular disease caused by VACV in dairy herds of six counties of Goiás state, Midwestern Brazil (2010-2012), involving a total of 122 cows, 12 calves and 11 people. Dairy cows (3 to 9 years old) were affected in all cases and calves (2 to 9 months old) were affected in five outbreaks, presenting oral lesions. The morbidity ranged between 8 and 100% in cows, and 1.5 to 31% in calves. In the cows, the clinical signs started with vesicles (2-7mm), painful and coalescent papules (3-8 mm), which resulted in ulcers (5-25mm) and scabs in teats, and, occasionally, in the muzzle. The clinical course lasted from 16 to 26 days. The histopathology of bovine skin samples revealed superficial perivascular inflammatory infiltrate of lymphocytes, plasma cells, neutrophils, macrophages and multifocal areas of acanthosis, spongiosis, hipergranulosis and parakeratotic or orthokeratotic hyperkeratosis with adjacent focally extensive ulcers. Eosinophilic inclusion bodies were noted in the cytoplasm of the keratinocytes. PCR to vgf gene of Orthopoxvirus was positive in samples collected from all outbreaks, and in some cases, genomic VACV sequences were identified by nucleotide sequencing of the PCR amplicons. Infectious virus was isolated in cell culture from scabs from one outbreak. Antibodies to Orthopoxvirus were detected in at least 3 or 4 animals in most outbreaks, by ELISA (outbreaks 1, 2, 3, 4, 5 and 7) or virus-neutralization (outbreak 6). Neutralizing titers ranging from 8 to 64 in outbreak 6. In all outbreaks, VACV infection was suspected based on the clinical and pathological findings and it was confirmed by laboratory tests. Upon the etiological confirmation, other agents associated with vesicular disease were discarded. In all outbreaks, at least one milker who handled the affected cows developed malaise, headache, fever, painful vesico-pustular lesions mainly in the hands, but also in the neck and nose. These results confirm the circulation of VACV in the region and call attention for a correct diagnosis and the adoption of prophylactic and control measures.

Effects of using different host plants on the detected biodiversity of arbuscular mycorrhizal fungi from an agroecosystem

The influence of peanut (Arachis hypogaea L.), sorghum (Sorghum bicolor (L.) Moench) and maize (Zea mays L.) on the development and diversity of arbuscular mycorrhizal fungi (AMF) from an agrosystem was investigated. Soil collected from an agricultural field where maize had been grown was inserted into sowing holes, under the seeds of peanut, sorghum and maize those were subsequently grown in sterilised quartz sand separately in plastic boxes for five months. After this period, collections of roots and rhizospheric soil were made to evaluate the percentages of root colonization (RC), number of spores (NS) and species of AMF. Peanut showed the highest average values for RC and NS: 24.5% and 547.8/100 g of soil, respectively. Maize had an average RC of 19.7% and 415.2 spores/100g soil. Sorghum showed the lowest values: 15.9% for average RC and 349.8 spores/100 g soil. A total of fourteen species of AMF were identified. Seven species were identified from peanut rhizospheres, Entrophospora colombiana being the most abundant and frequent. In sorghum rhizospheres, twelve species were found, Glomus geosporum was the dominant taxon in terms of number of spores and frequency. Ten species were detected in maize with Acaulospora longula being the most abundant and the most frequent. It was observed that peanut was the best plant for promoting the sporulation of AMF, while sorghum favoured the establishment of most AMF species, followed by maize.

Leaf anatomy variation within and between three "restinga" populations of Erythroxylum ovalifolium Peyr: (Erythroxylaceae) in Southeast Brazil

Erythroxylum ovalifolium is a woody shrub widespread in the "restinga", i.e. the open scrub vegetation of the Brazilian coastal sandy plains. We examined leaf anatomy variation of this species both within populations and between populations of three "restingas" in the state of Rio de Janeiro. Sites were ca.100 km far from each other and differed in regard to rainfall and vegetation structure: a dry, open site; a wet, dense site and an intermediate one. Microhabitats within sites were: (i) exposed to full irradiance, outside vegetation islands; (ii) partially exposed to full irradiance, at the border of vegetation islands; (iii) shaded, inside vegetation islands. Leaf anatomy parameters were measured for five leaves collected in each of five plants per microhabitat, in each population; they were thickness of the leaf blade, of the palisade and spongy parenchyma, and of the adaxial and abaxial epidermis. Leaves from the dry, open site had narrower abaxial epidermis and a smaller contribution of spongy parenchyma to total leaf blade thickeness than the other two sites, which we attributed to water stress. Adaxial epidermis and leaf are thicker in more exposed microhabitats (i and ii, above), irrespective of site. We proposed that between-site anatomical variation in traits related to water stress, and within-site anatomical variation in traits related to light-use are indicative of ecological plasticity and might help explain the high abundance of E. ovalifolium in the studied populations and along the State of Rio de Janeiro coast.

Is the infertility in hypothyroidism mainly due to ovarian or pituitary functional changes?

The objective of the present study was to examine whether hypothyroidism affects the reproductive system of adult female rats by evaluating ovarian morphology, uterus weight and the changes in serum and pituitary concentrations of prolactin and gonadotropins. Three-month-old female rats were divided into three groups: control (N = 10), hypothyroid (N = 10), treated with 0.05% 6-propyl-2-thiouracil (PTU) in drinking water for 60 days, and T4-treated group (N = 10), receiving daily sc injections of L-thyroxine (0.8 µg/100 g body weight) during the last 10 days of the experiment. At the end of 50 days of hypothyroidism no hypothyroid animal showed a regular cycle, while 71% of controls as well as the T4-treated rats showed regular cycles. Corpora lutea, growing follicles and mature Graafian follicles were found in all ovaries studied. The corpora lutea were smaller in both the hypothyroid and T4-replaced rats. Graafian follicles were found in 72% of controls and only in 34% of hypothyroid and 43% of T4-treated animals. Serum LH, FSH, progesterone and estradiol concentrations did not differ among the three groups. Serum prolactin concentration and the pituitary content of the three hormones studied were higher in the hypothyroid animals compared to control. T4 treatment restored serum prolactin concentration to the level found in controls, but only partially normalized the pituitary content of gonadotropins and prolactin. In conclusion, the morphological changes caused by hypothyroidism can be a consequence of higher prolactin production that can block the secretion and action of gonadotropins, being the main cause of the changes observed.

A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide

We report here the construction of a vector derived from pET3-His and pRSET plasmids for the expression and purification of recombinant proteins in Escherichia coli based on T7 phage RNA polymerase. The resulting pAE plasmid combined the advantages of both vectors: small size (pRSET), expression of a short 6XHis tag at N-terminus (pET3-His) and a high copy number of plasmid (pRSET). The small size of the vector (2.8 kb) and the high copy number/cell (200-250 copies) facilitate the subcloning and sequencing procedures when compared to the pET system (pET3-His, 4.6 kb and 40-50 copies) and also result in high level expression of recombinant proteins (20 mg purified protein/liter of culture). In addition, the vector pAE enables the expression of a fusion protein with a minimal amino-terminal hexa-histidine affinity tag (a tag of 9 amino acids using XhoI restriction enzyme for the 5'cloning site) as in the case of pET3-His plasmid and in contrast to proteins expressed by pRSET plasmids (a tag of 36 amino acids using BamHI restriction enzyme for the 5'cloning site). Thus, although proteins expressed by pRSET plasmids also have a hexa-histidine tag, the fusion peptide is much longer and may represent a problem for some recombinant proteins.

Age-related changes in radiation-induced micronuclei among healthy adults

The aim of the present study was to establish the extent of in vitro radioresponse of lymphocytes among 62 healthy adults of both genders and to estimate the distribution of baseline micronuclei and radiosensitivity among individuals of the study population using the cytochalasin block micronucleus test. A younger study group consisted of 10 males (mean age, 22.4 years; range, 21-27) and 12 females (mean age, 24.8 years; range, 20-29), whereas an older study group consisted of 18 males (mean age, 35.1 years; range, 30-44) and 22 females (mean age, 38.5 years; range, 30-48). For evaluation of radiosensitivity blood samples were irradiated in vitro using 60Co g-ray source. The radiation dose employed was 2 Gy, the dose rate 0.45 Gy/min. The study revealed a significant gender effect on baseline micronuclei favoring females (Z = 3.25, P < 0.001), while yields of radiation-induced micronuclei did not differ significantly (Z = 0.56, P < 0.56) between genders. The distribution of baseline micronuclei among the individuals tested followed Poisson distribution in both study groups and in both genders, whereas the distribution of radiosensitivity among individuals of the older study group did not fulfill Poisson expectations (Kolmogorov-Smirnof test, P < 0.01). In contrast to a nonsignificant difference in radiosensitivity between males and females of the same age group (Z = 1.97, P < 0.56), a statistically significant difference in radiosensitivity between younger and older group for both genders was found (Z = 3.03, P < 0.03). Since the individuals tested were healthy, the observed variability in radiation response is considered to be an early effect of ageing.

Patterns of intracellular cytokines in CD4 and CD8 T cells from patients with mycobacterial infections

Using a short-term bulk culture protocol designed for an intracellular-staining method based on a flow cytometry approach to the frequencies of cytokine-producing cells from tuberculosis and leprosy patients, we found distinct patterns of T cell subset expression. The method also reveals the profile of peak cytokine production and can provide simultaneous information about the phenotype of cytokine-producing cells, providing a reliable assay for monitoring the immunity of these patients. The immune response of Mycobacterium leprae and purified protein derivative (PPD) in vitro to a panel of mycobacteria-infected patients from an endemic area was assessed in primary mononuclear cell cultures. The kinetics and source of the cytokine pattern were measured at the single-cell level. IFN-gamma-, TNF-alpha-, IL-4- and IL-10-secreting T cells were intracytoplasmic evaluated in an attempt to identify M. leprae- and PPD-specific cells directly from the peripheral blood. The analysis by this approach indicated that TNF-alpha was the first (8 h) to be produced, followed by IFN-gamma (16 h), IL-10 (20 h) and IL-4 (24 h), and double-staining experiments confirmed that CD4+ were a greater source of TNF-alpha than of CD8+ T cells (P < 0.05). Both T cell subsets secreted similar amounts of IFN-gamma. We conclude that the protocol permits rapid evaluation of cytokine production by different T cell populations. The method can also be used to define immune status in non-infected and contact individuals.

Assessing negative priming by attended distractors in a paper-and-pencil task

The paper-and-pencil digit-comparison task for assessing negative priming (NP) was introduced, using a referent-size-selection procedure that was demonstrated to enhance the effect. NP is indicated by slower responses to recently ignored items, and proposed within the clinical-experimental framework as a major cognitive index of active suppression of distracting information, critical to executive functioning. The digit-comparison task requires circling digits of a list with digit-asterisk pairs (a baseline measure for digit-selection), and the larger of two digits in each pair of the unrelated (with different digits in successive digit-pairs) and related lists (in which the smaller digit subsequently became a target). A total of 56 students (18-38 years) participated in two experiments that explored practice effects across lists and demonstrated reliable NP, i.e., slowing to complete the related list relative to the unrelated list, (F(2, 44) = 52.42, P < 0.0001). A 3rd experiment examined age-related effects. In the paper-and-pencil digit-comparison task, NP was reliable for the younger (N = 8, 18-24 years) and middle-aged adults (N = 8, 31-54 years), but absent for the older group (N = 8, 68-77 years). NP was also reduced with aging in a computer-implemented digit-comparison task, and preserved in a task typically used to test location-specific NP, accounting for the dissociation between identity- and spatial-based suppression of distractors (Rao R(3, 12) = 16.02, P < 0.0002). Since the paper-and-pencil digit-comparison task can be administered easily, it can be useful for neuropsychologists seeking practical measures of NP that do not require cumbersome technical equipment.

Preliminary data on the prevalence of psychiatric disorders in Brazilian male and female juvenile delinquents

The aim of the present investigation was to study the prevalence of psychiatric disorders in a sample of delinquent adolescents of both genders and to compare the prevalence between genders. A total of 116 adolescents (99 males and 17 females) aged 12 to 19 on parole in the State of Rio de Janeiro were interviewed using the screening interview based on the Schedule for Affective Disorders and Schizophrenia for School-Age Children - Present and Lifetime (KSADS-PL). Data were collected between May 2002 and January 2003. Of 373 male and 58 female adolescents present in May 2002 in the largest institution that gives assistance to adolescents on parole in the city of Rio de Janeiro, 119 subjects were assessed (three of them refused to participate). Their average age was 16.5 years with no difference between genders. The screening interview was positive for psychopathology for most of the sample, with the frequencies of the suggested more prevalent psychiatric disorders being 54% for attention-deficit/hyperactivity disorder, 77% for conduct disorder, 41% for oppositional defiant disorder, 57% for anxiety disorder 57, 60% for depressive disorder 60, 63% for illicit drug abuse, and 58% for regular alcohol use. Internalizing disorders (depressive disorders, anxiety disorders and phobias) were more prevalent in the female subsample. There was no significant difference in the prevalence of illicit drug abuse between genders. There were more male than female adolescents on parole and failure to comply with the sentence was significantly more frequent in females. The high prevalence of psychopathology suggested by this study indicates the need for psychiatric treatment as part of the prevention of juvenile delinquency or as part of the sentence. However, treatment had never been available for 93% of the sample in this study.

Relative contribution of expectancy and immediate arousal to the facilitatory effect of an auditory accessory stimulus

An auditory stimulus speeds up a digital response to a subsequent visual stimulus. This facilitatory effect has been related to the expectancy and the immediate arousal that would be caused by the accessory stimulus. The present study examined the relative contribution of these two influences. In a first and a third experiment a simple reaction time task was used. In a second and fourth experiment a go/no-go reaction time task was used. In each of these experiments, the accessory stimulus preceded the target stimulus by 200 ms for one group of male and female volunteers (G Fix). For another group of similar volunteers (G Var) the accessory stimulus preceded the target stimulus by 200 ms in 25% of the trials, by 1000 ms in 25% of the trials and was not followed by the target stimulus in 50% of the trials (Experiments 1a and 1b) or preceded the target stimulus by 200 ms in 6% of the trials and by 1000 ms in 94% of the trials (Experiments 2a and 2b). There was a facilitatory effect of the accessory stimulus for G Fix in the four experiments. There was also a facilitatory effect of the accessory stimulus at the 200-ms stimulus onset asynchrony for G Var in Experiments 1a and 1b but not in Experiments 2a and 2b. The facilitatory effects observed were larger in the go/no-go task than in the simple task. Taken together, these results suggest that expectancy is much more important than immediate arousal for the improvement of performance caused by an accessory stimulus.