Dengue outbreaks: unpredictable incidence time series.

Dengue fever is a disease with increasing incidence, now occurring in some regions which were not previously affected. Ribeirão Preto and São Paulo, municipalities in São Paulo state, Brazil, have been highlighted due to the high dengue incidences especially after 2009 and 2013. Therefore, the current study aims to analyse the temporal behaviour of dengue cases in the both municipalities and forecast the number of disease cases in the out-of-sample period, using time series models, especially SARIMA model. We fitted SARIMA models, which satisfactorily meet the dengue incidence data collected in the municipalities of Ribeirão Preto and São Paulo. However, the out-of-sample forecast confidence intervals are very wide and this fact is usually omitted in several papers. Despite the high variability, health services can use these models in order to anticipate disease scenarios, however, one should interpret with prudence since the magnitude of the epidemic may be underestimated.

First detection and molecular characterization of Nebovirus in Brazil.

Nebovirus is a new genus of viruses belonging to the Caliciviridae family recently characterized in cattle, and is associated with gastrointestinal disorders, such as diarrhoea, anorexia and intestinal lesions particularly in calves. The aim of this study was to investigate the prevalence of neboviruses in Brazilian cattle and analyse phylogenetically the virus strains detected. A prevalence of 4·8% of neboviruses in faecal samples from 62 head of cattle from different Brazilian states was detected. All positive animals were aged 96·0% nt (100% aa) sequence identity between the virus sequences in this study and >88·8% nt (>94·4% aa) identity with Newbury1/UK. Our results indicate, for the first time, the occurrence of neboviruses in Brazil as well as in South America, and the first Newbury1-like nebovirus found outside the UK.

Controversial results of therapy with mesenchymal stem cells in the acute phase of canine distemper disease.

Distemper disease is an infectious disease reported in several species of domestic and wild carnivores. The high mortality rate of animals infected with canine distemper virus (CDV) treated with currently available therapies has driven the study of new efficacious treatments. Mesenchymal stem cell (MSC)-based therapy is a promising therapeutic option for many degenerative, hereditary, and inflammatory diseases. Therefore, the aim of this study was to characterize stem cells derived from the canine fetal olfactory epithelium and to assess the systemic response of animals infected with CDV to symptomatic therapy and treatment with MSCs. Eight domestic mongrel dogs (N = 8) were divided into two groups: support group (SG) (N = 5) and support group + cell therapy (SGCT) (N = 3), which were monitored over 15 days. Blood samples were collected on days 0, 6, 9, 12, and 15 to assess blood count and serum biochemistry (urea, creatinine, alanine transferase, alkaline phosphatase, gamma-glutamyl transferase, total protein, albumin, and globulin), and urine samples were obtained on days 0 and 15 for urinary evaluation (urine I). The results showed a high mortality rate (SG = 4 and SGCT = 2), providing inadequate data on the clinical course of CDV infection. MSC therapy resulted in no significant improvement when administered during the acute phase of canine distemper disease, and a prevalence of animals with high mortality rate was found in both groups due to the severity of symptoms.

Ki-67 antigen expression in the mammary epithelium of female rats in persistent estrus treated with raloxifene.

OBJECTIVE: To evaluate Ki-67 antigen expression in the mammary epithelium of female rats in persistent estrus treated with raloxifene. MATERIALS AND METHODS: Forty-one Wistar-Hannover rats in persistent estrus induced by 1.25 mg of testosterone propionate were randomly divided into two groups: Group A (control, n = 21) in which the animals received only the vehicle (propylene glycol) and Group B (experimental, n = 20) in which the rats received 750 µg/day of raloxifene by gavage. After 21 days of treatment, all the animals were sacrificed and the first pair of abdominal-inguinal mammary glands was extirpated and fixed in 10% buffered formalin to investigate Ki-67 expression by immunohistochemistry. The data were analysed using Student's t-test (p < 0.05). RESULTS: The percentage of Ki-67-stained nuclei per 500 cells in the mammary epithelium was 42.33 ± 6.18 and 15.51 ± 3.71 [mean ± standard error of the mean (SEM)] in the control and experimental groups, respectively (p < 0.001). CONCLUSION: Raloxifene treatment significantly reduced Ki-67 expression in the mammary epithelium of rats in persistent estrus.

Validation of two real-time PCRs targeting the PE-PGRS 20 gene and the region of difference 4 for the characterization of Mycobacterium bovis isolates.

This study aimed to develop and validate real-time PCR for the diagnosis of Mycobacterium bovis isolates. Two hundred and seventy-four M. bovis isolates and 156 M. tuberculosis isolates were tested. Both qPCRs amplified all of the 274 M. bovis samples, but none of the 156 M. tuberculosis samples. The qPCR for PE-PGRS 20 had 91% efficiency and a detection limit of 0.32 ng (sensitivity and specificity for qPCR "Mbovis.100" were 99.64 and 100%, respectively). The qPCR for RD4 had 100% efficiency, and a detection limit of 4 pg (diagnostic sensitivity and specificity were 100 and 100%. The qPCR tests were performed using 4 extraction sets, 3 qPCR kits, and with a range of equipment; yet, all combinations produced similar results in a diagnostic test, demonstrating the robustness of this method. The techniques proved to be efficient, robust, sensitive, and specific for the diagnosis of M. bovis.

Cerebrovascular risk factors and their time-dependent effects on stroke survival in the EMMA cohort study.

To investigate the time-dependent effects of traditional risk factors on functional disability in all-cause mortality post-stroke, we evaluated data from a long-term stroke cohort. Baseline cerebrovascular risk factors (CVRF) and functionality at 1 and 6 months were evaluated in survivors from a prospective stroke cohort using the modified Rankin scale (m-RS), which classifies participants as improvement of disability, unchanged disability (at least moderate), and worsening disability. Cox regression models considering baseline risk factors, medication use, and functionality 6 months after stroke were fitted to identify their time-dependent effects up to 12 years of follow-up. Adjusted hazard ratios (HR) with 95% confidence intervals (CI) are presented. Among 632 survivors (median age 68, 54% male, 71% first-ever episode), age and functional disability (unchanged and worsening) 6 months after ischemic stroke had time-dependent effects on all-cause mortality risk up to 12 years of follow-up. The most impacting risk factors were unchanged (at least moderate) (HR, 2.99; 95%CI: 1.98-4.52) and worsening disability (HR, 2.85; 95%CI: 1.26-6.44), particularly in the first two years after a stroke event (Time 1: ≥6 mo to <2.5 y). Worsening disability also impacted mortality in the period from ≥2.5 to <7.5 years (Time 2) of follow-up (HR, 2.43 (95%CI: 1.03-5.73). Other baseline factors had a fixed high-risk effect on mortality during follow-up. Post-stroke and continuous medication use had a fixed protective effect on mortality. Functional disability was the main contributor with differential risks of mortality up to 12 years of follow-up.

Effects of high-fat chow on heart tissue in acute and chronic experimental murine schistosomiasis mansoni.

This study aimed to investigate myocardial injuries in mice infected with Schistosoma mansoni and fed a high-fat chow. Sections of myocardial tissue from S. mansoni-infected mice, and controls that had been killed 9 and 17 weeks post-infection, were stained with H&E and Picrosirius red. Histopathological examination, stereological design-based method (optical disector) and morphometry (vessels, cardiomyocytes and an amount of collagen) were used. Data were analysed using two-way ANOVA. Regardless of time of infection, myocardial tissue from the infected mice fed high-fat chow showed myocarditis characterized by a higher number of inflammatory foci, several areas displaying coagulation of cardiac fibres, a greater loss of cardiomyocytes and fibroblast proliferation than in the standard chow control. Comparing infected mice from acute and chronic infections, a higher cardiomyocyte hyperplasia (P < 0·0001) and higher amounts of collagen (P < 0·05) were observed than in standard chow control. In addition, all animals fed high-fat chow showed lower numerical density and total number of cardiomyocytes (P < 0·05), thicker vessel walls and narrowed luminal intramyocardial vessels (P > 0·05) than in the standard chow control. Altogether the data supported the view that a double burden has a synergistic deleterious effect on the myocardial tissue.

Characterization of inflammatory cell infiltrate in human dental pulpitis.

INTRODUCTION: To evaluate the microscopic characteristics and densities (per mm(2) ) of tryptase(+) mast cells, CD4(+) T helper lymphocytes, CD45RO(+) memory T lymphocytes, foxp3(+) T regulatory lymphocytes, CD20(+) B lymphocytes, CD68(+) macrophages, and CD31(+) blood vessels in human dental pulpitis (n=38) and healthy pulpal tissue (n=6). METHODOLOGY: The pulps of 38 human teeth with a clinical diagnosis of irreversible pulpitis were removed by pulpectomy. The pulp tissue was immersed in 10% buffered formalin for evaluation using light microscopy. Tryptase, CD4, CD45RO, foxp3, CD20, CD68, and CD31 expressions were analysed using immunohistochemistry; other microscopic features, such as intensity of inflammatory infiltrate and collagen deposition, were evaluated using haematoxylin and eosin stain. Wilcoxon and Mann-Whitney tests were used for statistical analysis. The significance level was set at α=5%. RESULTS: Two microscopic patterns of pulpitis were found: group 1 (G1) (n=15) had an intense inflammatory infiltrate and mild collagen deposition; conversely, group 2 (G2) (n=23) had a scarce inflammatory infiltrate and intense collagen deposition. The numbers of CD68(+) macrophages (P=0.004) and CD20(+) B (P=0.068) lymphocytes and the density of blood vessels (P=0.002) were higher in G1 than in G2. However, a similar number of CD4(+) and CD45RO(+) T lymphocytes was found in both groups (P>0.05). When present, tryptase(+) mast cells were equally distributed in G1 and G2, whereas foxp3(+) T regulatory lymphocytes were detected in 59% and 14% of the samples of G1 and G2. Controls exhibited lower numbers of foxp3, tryptase, CD4, CD45RO, CD68 and CD20 positive cells than G1 and G2. CONCLUSIONS: Irreversible pulpitis had distinct microscopic features with important quantitative and qualitative differences in inflammatory cell infiltration.

Digital subtraction radiography evaluation of the bone repair process of chronic apical periodontitis after root canal treatment.

AIM: To monitor radiographically the progress of bone repair within chronic periapical lesions after root canal treatment using digital subtraction radiography (DSR). METHODOLOGY: Twelve patients with 17 single-rooted teeth with chronic apical periodontitis associated with an infected necrotic pulp were selected for root canal treatment. Periapical radiographs were taken before treatment (baseline) and immediately post-treatment, 45, 90, 135 and 180 days after treatment. The radiographic protocol included the use of individualized film holders with silicone bite blocks. The six radiographic images were digitized and submitted to digital subtraction using DSR software, resulting in five subtracted images (SI). Quantitative analysis of these SI was performed using Image Tool software to assess pixel value changes, considering a step-wedge as the gold standard and a cut-off value of 128 pixels. The aim was to identify any increase or decrease in mineral density in the region of the periapical lesion. RESULTS: A minor decrease in mineral density at the canal filling session and a significant progressive mineral gain in the following evaluations (P < 0.001) occurred. Pairwise comparison of pixel grey values revealed that only the 180-day follow-up differed significantly from the previous SI. CONCLUSION: Digital subtraction radiography is a useful method for evaluating the progress of bone repair after root canal treatment. Noticeable mineral gain was observed approximately 90 days after root canal filling and definite bone repair after 180 days.

Comparison of three vascular endothelial markers in the evaluation of microvessel density in breast cancer.

PURPOSE: To evaluate the microvessel density by comparing the performance of anti-factor VIII-related antigen, anti-CD31 and anti-CD34 monoclonal antibodies in breast cancer. METHODS: Twenty-three postmenopausal women diagnosed with Stage II breast cancer submitted to definitive surgical treatment were evaluated. The monoclonal antibodies used were anti-factor VIII, anti-CD31 and anti-CD34. Microvessels were counted in the areas of highest microvessel density in ten random fields (200 x). The data were analyzed using the Kruskal-Wallis nonparametric test (p < 0.05). RESULTS: Mean microvessel densities with anti-factor VIII, anti-CD31 and anti-CD34 were 4.16 +/- 0.38, 4.09 +/- 0.23 and 6.59 +/- 0.42, respectively. Microvessel density as assessed by anti-CD34 was significantly greater than that detected by anti-CD31 or anti-factor VIII (p < 0.0001). There was no statistically significant difference between anti-CD31 and anti-factor VIII (p = 0.4889). CONCLUSION: The density of stained microvessels was greater and staining was more intense with anti-CD34 compared to anti-CD31 and anti-factor VIII-related antigen.

Cerebrovascular risk factors and their time-dependent effects on stroke survival in the EMMA cohort study

To investigate the time-dependent effects of traditional risk factors on functional disability in all-cause mortality post-stroke, we evaluated data from a long-term stroke cohort. Baseline cerebrovascular risk factors (CVRF) and functionality at 1 and 6 months were evaluated in survivors from a prospective stroke cohort using the modified Rankin scale (m-RS), which classifies participants as improvement of disability, unchanged disability (at least moderate), and worsening disability. Cox regression models considering baseline risk factors, medication use, and functionality 6 months after stroke were fitted to identify their time-dependent effects up to 12 years of follow-up. Adjusted hazard ratios (HR) with 95% confidence intervals (CI) are presented. Among 632 survivors (median age 68, 54% male, 71% first-ever episode), age and functional disability (unchanged and worsening) 6 months after ischemic stroke had time-dependent effects on all-cause mortality risk up to 12 years of follow-up. The most impacting risk factors were unchanged (at least moderate) (HR, 2.99; 95%CI: 1.98-4.52) and worsening disability (HR, 2.85; 95%CI: 1.26-6.44), particularly in the first two years after a stroke event (Time 1: ≥6 mo to <2.5 y). Worsening disability also impacted mortality in the period from ≥2.5 to <7.5 years (Time 2) of follow-up (HR, 2.43 (95%CI: 1.03-5.73). Other baseline factors had a fixed high-risk effect on mortality during follow-up. Post-stroke and continuous medication use had a fixed protective effect on mortality. Functional disability was the main contributor with differential risks of mortality up to 12 years of follow-up.

Contested dominance modifies the anovulatory consequences of social subordination in female marmosets.

Dominance status among female marmosets is reflected in agonistic behavior and ovarian function. Socially dominant females receive submissive behavior from subordinates, while exhibiting normal ovulatory function. Subordinate females, however, receive agonistic behavior from dominants, while exhibiting reduced or absent ovulatory function. Such disparity in female fertility is not absolute, and groups with two breeding females have been described. The data reported here were obtained from 8 female-female pairs of captive female marmosets, each housed with a single unrelated male. Pairs were classified into two groups: "uncontested" dominance (UD) and "contested" dominance (CD), with 4 pairs each. Dominant females in UD pairs showed significantly higher frequencies (4.1) of agonism (piloerection, attack and chasing) than their subordinates (0.36), and agonistic behaviors were overall more frequently displayed by CD than by UD pairs. Subordinates in CD pairs exhibited more agonistic behavior (2.9) than subordinates in UD pairs (0.36), which displayed significantly more submissive (6.97) behaviors than their dominants (0.35). The data suggest that there is more than one kind of dominance relationship between female common marmosets. Assessment of progesterone levels showed that while subordinates in UD pairs appeared to be anovulatory, the degree of ovulatory disruption in subordinates of CD pairs was more varied and less complete. We suggest that such variation in female-female social dominance relationships and the associated variation in the degree and reliability of fertility suppression may explain variations of the reproductive condition of free-living groups of common marmosets.

Effect of protein supplementation on yield and milk composition of F1 Holstein x Zebu cows kept in deferred pasture of brachiaria grass / [Efeito da suplementação de proteína na produção e na composição do leite de F1 vacas Holandês x Zebu mantidas em pasto diferido de capim-braquiária]

The objective of this study was to evaluate different supplementation strategies concentrated to F1 Holstein x Zebu lactating cows managed in deferred signal grass pasture on the yield and composition of milk and body weight gain. Thirty six F1 Holstein x Zebu cows with an average lactation period of 267 ± 10 days, mean body weight of 548 ± 19kg were used following a completely randomized design in a 4 x 5 factorial scheme, being four feeding strategies and five days under evaluation. The treatments consisted of four nutritional strategies: deferred pasture as a source of roughage without supplementation (PDSS); deferred pasture as a source of roughage with protein supplement offer (PDCS); deferred pasture supplemented with 15 kilos of corn silage (natural base) + 1,200 grams of protein supplement (PDSP) and corn silage (ad libitum) + 700 grams of protein supplement (SMP). There was no interaction (P> 0.05) between the nutritional plans and days under evaluation for any of the variables. It was found that cows fed SMP showed milk production 26.06% higher than the other nutritional plans (mean of 11.46kg/day). F1 Holstein/Zebu cows handled in deferred pasture in a traditional way supplemented with protein maintains milk yield.(AU)

Objetivou-se avaliar diferentes planos nutricionais para vacas F1 Holandês/Zebu mantidas em pasto diferido de capim-braquiária sobre a produção e a composição do leite e no ganho em peso corporal. Foram utilizadas 36 vacas F1 Holandês/Zebu com período médio de lactação de 267 ± 10 dias, peso corporal médio de 548 ± 19kg, seguindo o delineamento inteiramente ao acaso, em esquema fatorial 4 x 5, sendo quatro estratégias de alimentação e cinco dias em avaliação. Os tratamentos consistiram de quatro planos nutricionais: pasto diferida como fonte de forragem sem suplementação (PDSS); pastagem diferida como fonte de forragem com oferta de suplemento de proteico (PDCS); pasto diferido suplementado com 15 quilos de silagem de milho (base natural) + 1.200 gramas de suplemento proteico (PDSP) e silagem de milho (ad libitum) + 700 gramas de suplemento de proteína (SMP). Verificou-se que as vacas alimentadas com SMP apresentaram produção de leite 26,06% superior aos demais planos nutricionais (média de 11,46kg/dia). Vacas F1 Holandês/Zebu tratadas em pastagem diferida de maneira tradicional, suplementada com proteína, mantêm o rendimento de leite.(AU)

Experimental paracoccidioidomycosis in immunosuppressed mice.

Mice were immunosuppressed by means of wholebody irradiation or cyclophosphamide, in order to investigate the influence on the initial phase of infection induced by a strain of the fungus, Paracoccidioides brasiliensis, in the yeast phase and inoculated intraperitoneally. A group of mice was irradiated with 600 rad (cobalt gamma-irradiation) 24 h before infection. Two groups were treated with cyclophosphamide (200 mg/kg intravenously), one two days before, and the other, one day after infection. A control group received the fungus, but no radiation or cyclophosphamide. All animals developed lesions at the site of inoculation. Metastatic lesions were observed in 100% of the animals in the irradiated group, 67% in each of the cyclophosphamide-treated groups and 33% in the control group. These lesions were found both in the liver and lungs, being more numerous in the irradiated group, followed by the cyclophosphamide-treated group in which the drug was given after the infection; they were slight in both viscera in the other cyclophosphamide-treated group and also slight in the liver and absent in the lungs of the controls.

Size distribution of recruited alveolar volumes in airway reopening.

In 11 isolated dog lung lobes, we studied the size distribution of recruited alveolar volumes that become available for gas exchange during inflation from the collapsed state. Three catheters were wedged into 2-mm-diameter airways at total lung capacity. Small-amplitude pseudorandom pressure oscillations between 1 and 47 Hz were led into the catheters, and the input impedances of the regions subtended by the catheters were continuously recorded using a wave tube technique during inflation from -5 cm H(2)O transpulmonary pressure to total lung capacity. The impedance data were fit with a model to obtain regional tissue elastance (Eti) as a function of inflation. First, Eti was high and decreased in discrete jumps as more groups of alveoli were recruited. By assuming that the number of opened alveoli is inversely proportional to Eti, we calculated from the jumps in Eti the distribution of the discrete increments in the number of opened alveoli. This distribution was in good agreement with model simulations in which airways open in cascade or avalanches. Implications for mechanical ventilation may be found in these results.

Acoustic evidence of airway opening during recruitment in excised dog lungs.

The aim of this study was to test the hypothesis that the mechanism of recruitment and the lower knee of the pressure-volume curve in the normal lung are primarily determined by airway reopenings via avalanches rather than simple alveolar recruitments. In isolated dog lung lobes, the pressure-volume loops were measured, and crackle sounds were recorded intrabronchially during both the first inflation from the collapsed state to total lobe capacity and a second inflation without prior degassing. The inflation flow contained transients that were accompanied by a series of crackles. Discrete volume increments were estimated from the flow transients, and the energy levels of the corresponding crackles were calculated from the sound recordings. Crackles were concentrated in the early phase of inflation, with the cumulative energy exceeding 90% of its final value by the lower knee of the pressure-volume curve. The values of volume increments were correlated with crackle energy during the flow transient for both the first and the second inflations (r(2) = 0.29-0.73 and 0.68-0.82, respectively). Because the distribution of volume increments followed a power law, the correlation between crackle energy and discrete volume increments suggests that an avalanche-like airway opening process governs the recruitment of collapsed normal lungs.

Amyloidosis in experimental tegumentary leishmaniasis in mice.

Renal and hepatosplenic amyloidosis was found in chronic cutaneous leishmaniasis in mice infected with 10(6) purified amastigotes from lesions produced by the H21 strain of Leishmania mexicana amazonensis. After 1 year a progressive lesion leading to metastasis was observed in most animals.

Determination of the p-monochlorophenol residue in the calcium hydroxide + P-monochlorophenol combination used as an intracanal dressing in pulpless teeth of dogs with induced chronic periapical lesion.

To determine the presence of p-monochlorophenol in the calcium hydroxide (Calen) + p-monochlorophenol combination after its use as intracanal dressing, periapical lesions were induced in 60 root canals of upper and lower premolars of four dogs. After biomechanical preparation, the root canals received the intracanal medication, which was removed from the apical third after 2, 4, 7, and 14 days for chemical analysis by spectrophotometry. The results showed a p-monochlorophenol loss of approximately 50.0% in the dressing after 48 h, with no further significant loss after longer periods of times. p-Monochlorophenol was still present in the medication after 14 days.

Behavior and progesterone levels in Callithrix jacchus females.

In callitrichids, and especially in the Callithrix and Saguinus genera, reproductive suppression maintains the breeding exclusivity of dominant females. It has been suggested that dominance may be inferred from agonistic behavior of females. The purpose of this study was to assess the adequacy of behaviors as indicators of hormonal status in Callithrix jacchus females. Two pairs of twin females were observed for affiliative, agonistic and sexual behaviors for 20 weeks, in four stages of increasingly potential competition for one male. During the same period, blood samples were taken regularly for progesterone determination by ELISA. The two pairs differed markedly as to behaviors and hormonal status: pair 1 showed a well established behavioral dominance and only one ovulating female from the beginning of the observation period; pair 2 showed competition for dominance until the last stage of observation, when one of the females started ovulating. Correlations between behaviors and progesterone levels showed differences between subordinate and dominant females mostly in pair 2. Although limited in number of animals studied, these results suggest that there is a relationship between behavior and hormones in female marmosets, but this relationship may be useful in the identification of the physiological status of females only in groups where the dominance is not well established.