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Nuclear proliferation marker MIB-1 (Ki-67) immunohistochemistry (IHC) is used to examine tumor cell proliferation. However, the diagnostic or prognostic value of the Ki-67 nuclear staining intensity and location, defined as nuclear gradient (NG), has not been assessed. This study examined the potential association between Ki-67 NG and cell cycle phases and its effect on the prognosis of pulmonary typical carcinoid (PTC) tumors. We propose a method for classifying the NG of Ki-67 during the cell cycle and compare the results between PTC, pulmonary adenocarcinoma (PAD), and breast ductal carcinoma (BDC). A literature review and objective analysis of IHC-stained paraffin sections were used to determine the Ki-67 labeling index and composed a stratification of the NG into NG1, NG2, and NG3/4 categories. A semi-automated image analysis protocol was established to determine the Ki-67 NG in PTC, PAD, and BDC. High intraobserver consistency and moderate interobserver agreement were achieved in the determination of Ki-67 NG in tumor specimens. NG1 and NG2 were lower in PTC than in PAD and BDC. Cox multivariate analysis of PTC after adjusting for age and number of metastatic lymph nodes showed that Ki-67 NG1 and NG2 significantly predicted clinical outcomes. The semi-automated method for quantification of Ki-67 nuclear immunostaining proposed in this study could become a valuable diagnostic and prognostic tool in PTC.

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Nuclear proliferation marker MIB-1 (Ki-67) immunohistochemistry (IHC) is used to examine tumor cell proliferation. However, the diagnostic or prognostic value of the Ki-67 nuclear staining intensity and location, defined as nuclear gradient (NG), has not been assessed. This study examined the potential association between Ki-67 NG and cell cycle phases and its effect on the prognosis of pulmonary typical carcinoid (PTC) tumors. We propose a method for classifying the NG of Ki-67 during the cell cycle and compare the results between PTC, pulmonary adenocarcinoma (PAD), and breast ductal carcinoma (BDC). A literature review and objective analysis of IHC-stained paraffin sections were used to determine the Ki-67 labeling index and composed a stratification of the NG into NG1, NG2, and NG3/4 categories. A semi-automated image analysis protocol was established to determine the Ki-67 NG in PTC, PAD, and BDC. High intraobserver consistency and moderate interobserver agreement were achieved in the determination of Ki-67 NG in tumor specimens. NG1 and NG2 were lower in PTC than in PAD and BDC. Cox multivariate analysis of PTC after adjusting for age and number of metastatic lymph nodes showed that Ki-67 NG1 and NG2 significantly predicted clinical outcomes. The semi-automated method for quantification of Ki-67 nuclear immunostaining proposed in this study could become a valuable diagnostic and prognostic tool in PTC.

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Knowledge of plant responses to environmental heterogeneity during ontogeny is important to elucidate the changes that occur to promote resource capture in tropical forests. We tested the hypothesis that expression changes in leaf metamer traits of Brazilian rosewood (Dalbergia nigra), from seedlings to emergent canopy trees, occur as new microclimate environments are achieved. We also tested the hypothesis that increased light heterogeneity in the understorey leads to higher plasticity in leaf traits of seedlings and saplings than in sun-exposed metamers of emergent trees subject to stressful conditions. We compared leaf metamer traits of 53 individuals including seedlings, saplings and emergent trees. We also evaluated the light heterogeneity in vertical strata and the variations in leaf traits within individuals (among metamers of the same individual). These were associated with height of the individuals. Compared to understorey plants, emergent trees presented larger metamers, with lower specific leaf area (SLA), lower investment in leaf area per total dry mass of metamer (LARm ), lower specific petiole length (SPL) and lower specific internode length (SIL). Higher phenotypic variation within individuals was observed in seedlings, which decreased as the trees grew taller. The results suggest the integration of ontogenetic changes in leaf traits under new microclimate conditions as the plants reach different vertical strata in the forest. Additionally, our results support the hypothesis that increased light heterogeneity in the understorey shaped higher phenotypic variation within individuals in juveniles and that stressful conditions in sun-exposed leaf metamers of emergent trees led to increased phenotypic stability.

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Baculoviruses have been applied for biocontrol of agricultural pests, such as velvetbean caterpillar (Anticarsia gemmatalis) and fall armyworm (Spodoptera frugiperda). Cell culture is an interesting approach for large-scale production of these viruses. Co-infection of a host cell with two distinct viruses can contribute to reduce costs due to saving cell culture media, bioreactor space and the resulting co-occluded polyhedra may help to reduce final biopesticide costs. The baculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) and Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) were chosen to test a model for in vitro co-infection in SF21 cells. Different proportions of SfMNPV/AgMNPV were evaluated along three in vitro passages by optical microscopy analysis of cells and real-time PCR (qPCR) of DNA obtained from budded viruses (BVs) and occlusion bodies (OBs). The kinetics of viral protein synthesis was carried out for analysis of the co-infection in first passage and bioassays with the resulting OBs were performed against A. gemmatalis and S. frugiperda larvae. The results demonstrated successful co-infection in these cells. The quantity of SfMNPV and AgMNPV in supernatants and sediments tends to be maintained stable during the three passages, although the amount of AgMNPV was higher than SfMPNV in most of the experiments. Analysis of the kinetics of radiolabed proteins showed that the cell protein synthesis was shut off and two distinct bands of about 30 kDa, regarded to be the polyhedrin of each virus, were strongly detected at 48 and 72 hp.i. Although the pathogenicity of the produced viruses was not completely satisfactory, the bioassays confirmed occurrence of co-infected larvae with disproportional amount of each virus.

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Baculovirus natural populations are known to be genetically heterogeneous and such genotypic diversity could have implications in the performance of biocontrol agents. The Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) has been widely used to control the velvetbean caterpillar, Anticarsia gemmatalis, in Brazil. In the present work, morphological and molecular analyses as well as the biological activity of AgMNPV genotypes derived from a Brazilian field isolate (AgMNPV-79) were carried out. The existence of genotypic variants in the population was confirmed by DNA restriction analysis. Although difference in virulence was observed among the variants, the most (Ag79-01) and the least (AgL-16) virulent clones do not show any morphological and cytopathological changes when compared to the most studied isolate (AgMNPV-2D). The complete genome analysis of the two viral clones showed the presence of single open reading frames (ORFs) of the pe-38 and he65 genes, which contrasts with the two split ORFs present in the genome of the AgMNPV-2D isolate. The viral clone AgL-16 has many variations in the ie-2 and pe-38 genes, which are transcription regulatory genes responsible for the regulation of viral early gene expression during insect cell infection. Furthermore, other genes showed alterations like the odv-e56, which have an essential role in the maturation and envelopment of the ODVs, and bro-a and bro-b genes which were fused to form a single ORF. For the Ag79-01, although the total number of single nucleotide variants (SNVs) was more prominent in the pe-38 gene, its genome showed very few modifications in comparison to the AgMNPV-2D genome.

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The maintenance of wild animals in captivity can be a source for transmission of infectious and zoonotic diseases. In February 2016, blue-fronted amazon parrots that were kept at the Centro de Reabilitação de Animais Silvestres (CRAS) of Campo Grande - MS died suddenly. The specimens were sent to the Universidade Federal de Mato Grosso do Sul (UFMS), Faculdade de Medicina Veterinária e Zootecnia (FAMEZ) to be necropsied. Anatomopathological exams were performed and organ fragments from all systems were collected for histopathological exams. Liver and spleen fragments were also collected for bacteriological exams. Histopathological exam revealed bacterial aggregates distributed through the organs. Bacteriological exam isolated Salmonella spp. from the fragments of liver and spleen. The confirmation of the genus was made by serum agglutination on slide and by the polymerase chain reaction (PCR). The genetic sequencing identified the sample as Salmonella Typhimurium.(AU)

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The maintenance of wild animals in captivity can be a source for transmission of infectious and zoonotic diseases. In February 2016, blue-fronted amazon parrots that were kept at the Centro de Reabilitação de Animais Silvestres (CRAS) of Campo Grande - MS died suddenly. The specimens were sent to the Universidade Federal de Mato Grosso do Sul (UFMS), Faculdade de Medicina Veterinária e Zootecnia (FAMEZ) to be necropsied. Anatomopathological exams were performed and organ fragments from all systems were collected for histopathological exams. Liver and spleen fragments were also collected for bacteriological exams. Histopathological exam revealed bacterial aggregates distributed through the organs. Bacteriological exam isolated Salmonella spp. from the fragments of liver and spleen. The confirmation of the genus was made by serum agglutination on slide and by the polymerase chain reaction (PCR). The genetic sequencing identified the sample as Salmonella Typhimurium.(AU)

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OBJECTIVE: To compare the efficacy of a new dentifrice containing 1.5% arginine and 1450 ppm fluoride to a positive control dentifrice containing 1450 ppm fluoride alone in arresting and reversing primary root caries lesions in adults. STUDY DESIGN: A total of 3779 subjects from Piracicaba, Säo Paulo, Brazil were screened; 284 had at least one leathery primary root caries lesion and were eligible for the study. The new dentifrice contained 1.5% arginine, an insoluble calcium compound, and 1450 ppm fluoride, as sodium monofluorophosphate; the matched positive control dentifrice contained 1450 ppm fluoride. One lesion for each subject was selected for inclusion in the study and was examined at baseline, 3 and 6 months. RESULTS: A total of 253 subjects completed the study with 129 of 144 subjects included in the final statistical analysis for the test dentifrice and 124 of 140 for the positive control. The mean age of subjects was 45.7 (±9.19) years and 56.5% were female. After 6 months product use, 70.5% of root caries lesions improved for subjects using the arginine-containing dentifrice compared to 58.1% for subjects using the positive control. The difference in the number of root caries lesions becoming hard in the two groups was statistically significant (p=0.038). CONCLUSION: A new dentifrice containing 1.5% arginine, an insoluble calcium compound, and 1450 ppm fluoride, as sodium monofluorophosphate, provided statistically significantly superior efficacy in arresting and reversing active root caries lesions in adults compared to a matched positive control dentifrice containing fluoride alone.

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In a benthic community on a continuous flat granite substrate in a third-order coastal forest stream, the dominant chironomid (Cricotopus) increased in number when shrimps (Macrobrachium olfersi and Potimirim glabra) and baetid ephemeropterans were excluded by electricity. The response appeared to be mediated by an increase in periphyton and sediments, rather than a reduction of direct predation or interference. Chironomids, periphyton and sediments decreased significantly compared to the control when shrimps only were excluded. Baetid ephemeropteran appeared to be the most important determinants of periphyton and sediment mass; the density of chironomids appeared to follow the quantity of periphyton and sediments.

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