PSMA-PET and micro-ultrasound potential in the diagnostic pathway of prostate cancer.

PURPOSE: To compare the diagnostic performance of 68Ga-PSMA PET/TC with PRI-MUS (prostate risk identification using micro-ultrasound) in the primary diagnosis of prostate cancer (PCa). METHODS: From September till December 2018, we prospectively enrolled 25 candidates to 68Ga-PSMA PET/TRUS (transrectal ultrasound) fusion biopsy and compared them with PRI-MUS. This included patients with persistently elevated PSA and/or PHI (prostate health index) suspicious for PCa, negative digital rectal examination, with either negative or contraindication to mpMRI, and at least one negative biopsy. The diagnostic performance of the two modalities was calculated based on pathology results. RESULTS: Overall, 20 patients were addressed to 68Ga-PSMA PET/TRUS fusion biopsy. Mean SUVmax and SUVratio for PCa lesions resulted significantly higher than in benign lesions (p = 0.041 and 0.011, respectively). Using optimal cut-off points, 68Ga-PSMA PET/CT demonstrated an overall accuracy of 83% for SUVmax ≥ 5.4 and 94% for SUVratio ≥ 2.2 in the detection of clinically significant PCa (GS ≥ 7). On counterpart, PRI-MUS results were: score 3 in nine patients (45%), score 4 in ten patients (50%), and one patient with score 5. PRI-MUS score 4 and 5 demonstrated an overall accuracy of 61% in detecting clinically significant PCa. CONCLUSION: In this highly-selected patient population, in comparison to PRI-MUS, 68Ga-PSMA PET/CT shows a higher diagnostic performance.

Lianas, tree ferns and understory species: indicators of conservation status in the Brazilian Atlantic Rainforest remnants, southeastern Brazil / Lianas, fetos arborescentes e espécies de sub-bosque: indicadores do estado de conservação da Mata Atlântica brasileira

Abstract Indicators are applied faster and at lower costs than conventional surveys, providing quick and efficient responses that can facilitate protected areas management. Our aim was to select indicators from vegetation to monitor protected areas. For this purpose, we analyzed understory and quantified lianas and tree ferns in protected and non-protected areas, in order to find indicator species. Our study areas are located in Vale do Ribeira, southeastern São Paulo state, Brazil. One of the areas is under two protection categories (IUCN's categories II and V), and the other is a privately owned farm. Lianas with large diameters (> 13 cm) and tree ferns with great heights (> 19 m) were considered indicators of undisturbed areas (protected areas) because their growth is directly related to forest successional stage. Indicator species within the protected area were shade tolerant species, such as Bathysa australis (A.St.-Hil.) K.Schum., whereas outside the protected area were pioneer species, such as Pera glabrata (Schott) Poepp. ex Baill. e Nectandra oppositifolia Ness. All of the suggested indicators can be used in management actions, especially in protected areas, to guarantee forest maintenance and ensure fulfillment of the conservation objectives of these areas.

Resumo Os indicadores são aplicados com menores custos e mais rapidez em comparação com levantamentos convencionais, fornecendo respostas rápidas e eficientes que podem facilitar a gestão das áreas protegidas. Nosso objetivo foi selecionar indicadores de vegetação para monitorar áreas protegidas. Para este fim, analisamos o sub-bosque e quantificamos as lianas e fetos arborescentes em áreas protegidas e não-protegidas em busca de espécies indicadoras. Nossas áreas de estudo estão localizadas na região do Vale do Ribeira, sudeste do Estado de São Paulo, Brasil. Uma delas é uma área protegida (categorias da IUCN II e V) e outra uma fazenda de propriedade privada. Lianas com grandes diâmetros (>13 cm) e fetos arborescentes com grandes alturas (>19 m) foram considerados indicadores de áreas não perturbadas (área protegida), pois seu crescimento está diretamente associado com o estágio sucessional das florestas. As espécies indicadoras da área protegida são não pioneiras, como Bathysa australis (A.St.-Hil.) K.Schum., já as da fazenda (não protegida) são pioneiras, como Pera glabrata (Schott) Poepp. ex Baill. e Nectandra oppositifolia Ness. Todos indicadores sugeridos podem ser utilizados em ações de gestão e conservação, especialmente em áreas protegidas, para garantir a manutenção dos remanescentes florestais e para assegurar o cumprimento dos objetivos destas áreas.

Lianas, tree ferns and understory species: indicators of conservation status in the Brazilian Atlantic Rainforest remnants, southeastern Brazil / Lianas, fetos arborescentes e espécies de sub-bosque: indicadores do estado de conservação da Mata Atlântica brasileira

Indicators are applied faster and at lower costs than conventional surveys, providing quick and efficient responses that can facilitate protected areas management. Our aim was to select indicators from vegetation to monitor protected areas. For this purpose, we analyzed understory and quantified lianas and tree ferns in protected and non-protected areas, in order to find indicator species. Our study areas are located in Vale do Ribeira, southeastern Sمo Paulo state, Brazil. One of the areas is under two protection categories (IUCN's categories II and V), and the other is a privately owned farm. Lianas with large diameters (> 13 cm) and tree ferns with great heights (> 19 m) were considered indicators of undisturbed areas (protected areas) because their growth is directly related to forest successional stage. Indicator species within the protected area were shade tolerant species, such as Bathysa australis (A.St.-Hil.) K.Schum., whereas outside the protected area were pioneer species, such as Pera glabrata (Schott) Poepp. ex Baill. e Nectandra oppositifolia Ness. All of the suggested indicators can be used in management actions, especially in protected areas, to guarantee forest maintenance and ensure fulfillment of the conservation objectives of these areas.(AU)

Os indicadores são aplicados com menores custos e mais rapidez em comparação com levantamentos convencionais, fornecendo respostas rápidas e eficientes que podem facilitar a gestão das áreas protegidas. Nosso objetivo foi selecionar indicadores de vegetação para monitorar áreas protegidas. Para este fim, analisamos o sub-bosque e quantificamos as lianas e fetos arborescentes em áreas protegidas e não-protegidas em busca de espécies indicadoras. Nossas áreas de estudo estão localizadas na região do Vale do Ribeira, sudeste do Estado de São Paulo, Brasil. Uma delas é uma área protegida (categorias da IUCN II e V) e outra uma fazenda de propriedade privada. Lianas com grandes diâmetros (>13 cm) e fetos arborescentes com grandes alturas (>19 m) foram considerados indicadores de áreas não perturbadas (área protegida), pois seu crescimento está diretamente associado com o estágio sucessional das florestas. As espécies indicadoras da área protegida são não pioneiras, como Bathysa australis (A.St.-Hil.) K.Schum., já as da fazenda (não protegida) são pioneiras, como Pera glabrata (Schott) Poepp. ex Baill. e Nectandra oppositifolia Ness. Todos indicadores sugeridos podem ser utilizados em ações de gestão e conservação, especialmente em áreas protegidas, para garantir a manutenção dos remanescentes florestais e para assegurar o cumprimento dos objetivos destas áreas.(AU)

Lianas, tree ferns and understory species: indicators of conservation status in the Brazilian Atlantic Rainforest remnants, southeastern Brazil.

Indicators are applied faster and at lower costs than conventional surveys, providing quick and efficient responses that can facilitate protected areas management. Our aim was to select indicators from vegetation to monitor protected areas. For this purpose, we analyzed understory and quantified lianas and tree ferns in protected and non-protected areas, in order to find indicator species. Our study areas are located in Vale do Ribeira, southeastern São Paulo state, Brazil. One of the areas is under two protection categories (IUCN's categories II and V), and the other is a privately owned farm. Lianas with large diameters (> 13 cm) and tree ferns with great heights (> 19 m) were considered indicators of undisturbed areas (protected areas) because their growth is directly related to forest successional stage. Indicator species within the protected area were shade tolerant species, such as Bathysa australis (A.St.-Hil.) K.Schum., whereas outside the protected area were pioneer species, such as Pera glabrata (Schott) Poepp. ex Baill. e Nectandra oppositifolia Ness. All of the suggested indicators can be used in management actions, especially in protected areas, to guarantee forest maintenance and ensure fulfillment of the conservation objectives of these areas.

Antigenicity and immunogenicity of rotavirus VP6 protein expressed on the surface of Lactococcus lactis.

Group A rotaviruses are the major etiologic agents of acute gastroenteritis worldwide in children and young animals. Among its structural proteins, VP6 is the most immunogenic and is highly conserved within this group. Lactococcus lactis is a food-grade, Gram-positive, and nonpathogenic lactic acid bacteria that has already been explored as a mucosal delivery system of heterologous antigens. In this work, the nisin-controlled expression system was used to display the VP6 protein at the cell surface of L. lactis. Conditions for optimal gene expression were established by testing different nisin concentrations, cell density at induction, and incubation times after induction. Cytoplasmic and cell wall protein extracts were analyzed by Western blot and surface expression was confirmed by flow cytometry. Both analysis provided evidence that VP6 was efficiently expressed and displayed on the cell surface of L. lactis. Furthermore, the humoral response of mice immunized with recombinant L. lactis was evaluated and the displayed recombinant VP6 protein proved to be immunogenic. In conclusion, this is the first report of displaying VP6 protein on the surface of L. lactis to induce a specific immune response against rotavirus. These results provide the basis for further evaluation of this VP6-displaying L. lactis as a mucosal delivery vector in a mouse model of rotavirus infection.

Correlation between rotavirus A genotypes detected in hospitalized children and sewage samples in 2006, Córdoba, Argentina.

Routine rotavirus A (RV-A) surveillance is based on clinical cases, so only symptomatic infections are reported. The objective of this study was to determine whether the RV-A genotypes and cold seasonal pattern described in patients with diarrhea is reflected by sewage surveillance, which could be representative of the RV-A genotypes circulating in the population. The genotype distribution of RV-A in effluent samples from a local sewage treatment plant was compared to those from local clinical cases. A total of 52 sewage samples and 70 stool specimens from children with acute non-bacterial diarrhea were collected from January to December 2006. The effluent specimens were concentrated and RNA extracts from concentrated sewage and clinical samples were genotyped for the rotavirus VP7 gene. The proportional distribution of the RV-A G-genotypes in sewage and clinical samples during the cold season was similar: G1 accounted for 26.6% of the typed sewage isolates and 28.8% of the clinical infections; G3 type accounted for 21.9% and 25.8%; G2 type 15.6% and 10.6%; G4 type 17.2% and 21.2%; G8 type 1.6% and 0%; and the G9 type 17.2% and 13.6%, respectively. A similar picture of RV-A genotype detection was obtained in sewage samples collected during the cold and warm seasons. The results indicate that there is a correlation between genotypes of RV-A isolates from human diarrheic patients and of those from sewage samples. In addition, sewage monitoring highlighted the uniform all-year RV-A circulation, which was in contrast to the peak incidence of RV-A infection in the community.

Rotavirus VP7-gene selection during coinfections in CaCo-2 cells.

A previous rotavirus epidemiological survey in Córdoba, Argentina, revealed an unusually high frequency of mixed G-type infections (41.5%). The genotype distribution of those mixed infections showed that the most prevalent G-type combinations were G1+G4 (65.0%), G1+G2 (12.5%), G2+G4 (3.1%) and G1+G9 (2.5%). In the present study we analyzed the competitive growth in CaCo-2 cell cultures of strains from the most frequent rotavirus G-type coinfections in order to explain some aspect of the dynamic of G-type replacement along the time. Our results indicated that G1-type was preferentially selected compared with G2 and G9-genotypes, meanwhile, G1-G4 coinfections showed an efficient co-amplification of both types. Interestingly, this mirrored the high detection rates of both genotypes as single and mixed infections (G1+G4, 65.0%) in our region. On the other hand, G2-type revealed a better amplification rate with respect to G4-type. Fluctuant rates in the prevalence of different genotypes usually observed along the time could, in part, be explained by successive replacement of strains with different growth characteristics. We hypothesized that one aspect of these different fitnesses can be measured as differential growth in culture of the strains contained in the sample of a mixed infection. Our findings here provide the first data supporting the validity of the competitive replication in vitro to better understand rotavirus G-type circulation patterns.

Irrigação do timo em aves da linhagem Paraíso Pedrês (Gallus gallus domesticus) / Irrigation of thymus in Paraiso Pedres lineage (Gallus gallus domesticus) birds

A vascularização arterial do timo (número, origem e ordenação) bem como a distribuição parenquimal dos ramos penetrantes foram estudadas em 40 aves da linhagem Paraíso Pedrês. Trinta aves foram injetadas com látex e dissecadas, enquanto 10 aves tiveram seus sistemas arteriais injetados com resina (metil metacrilato e mercox) para a preparação de moldes vasculares. A principal fonte de irrigação encontrada foi a associação dos ramos oriundos das artérias comuns do nervo vago, tireóideas e ingluviais, sendo que seus ramos penetravam o parênquima dos lobos, principalmente pelas suas extremidades cranial e caudal. A partir da penetração, os ramos tímicos apresentavam distribuição predominante para a periferia do lobo, formando uma trama capilar poligonal, com espaços irregulares, característica de um órgão linforreticular.(AU)

The thymus arterial vascularization (number, origin and organization) as well as the parenchymal distribution of the penetrative branches were studied in 40 birds of Paraiso Pedres lineage birds. From this total, 30 were injected with latex and dissected, while 10 had their arterial vessels injected with resin (methylmethacrylate and mercox) for the vascular matrix preparation. The main source of irrigation found was the association of branches originated from the common arteries of the vago nerve, thyroid and ingluvial, and their branches penetrated in the parenchyma of the lobes especially through the caudal and cranial endings. From this penetration, the thymic branches showed most of their distribution to the lobe periphery, creating a polygonal capilar web with irregular, spaces which is a characteristic of a linforeticular organ.(AU)

Irrigação do timo em aves da linhagem Paraíso Pedrês (Gallus gallus domesticus) / Irrigation of thymus in Paraiso Pedres lineage (Gallus gallus domesticus) birds

A vascularização arterial do timo (número, origem e ordenação) bem como a distribuição parenquimal dos ramos penetrantes foram estudadas em 40 aves da linhagem Paraíso Pedrês. Trinta aves foram injetadas com látex e dissecadas, enquanto 10 aves tiveram seus sistemas arteriais injetados com resina (metil metacrilato e mercox) para a preparação de moldes vasculares. A principal fonte de irrigação encontrada foi a associação dos ramos oriundos das artérias comuns do nervo vago, tireóideas e ingluviais, sendo que seus ramos penetravam o parênquima dos lobos, principalmente pelas suas extremidades cranial e caudal. A partir da penetração, os ramos tímicos apresentavam distribuição predominante para a periferia do lobo, formando uma trama capilar poligonal, com espaços irregulares, característica de um órgão linforreticular.

The thymus arterial vascularization (number, origin and organization) as well as the parenchymal distribution of the penetrative branches were studied in 40 birds of Paraiso Pedres lineage birds. From this total, 30 were injected with latex and dissected, while 10 had their arterial vessels injected with resin (methylmethacrylate and mercox) for the vascular matrix preparation. The main source of irrigation found was the association of branches originated from the common arteries of the vago nerve, thyroid and ingluvial, and their branches penetrated in the parenchyma of the lobes especially through the caudal and cranial endings. From this penetration, the thymic branches showed most of their distribution to the lobe periphery, creating a polygonal capilar web with irregular, spaces which is a characteristic of a linforeticular organ.

Molecular epidemiology of group A rotavirus diarrhea among children in Buenos Aires, Argentina, from 1999 to 2003 and emergence of the infrequent genotype G12.

To examine the epidemiology of rotaviruses in Buenos Aires, Argentina, we screened 1,212 stool samples from children with diarrhea in the southern district of Buenos Aires from 1999 to 2003. We identified 187 samples (15.4%) that were positive for group A rotavirus by use of antigen enzyme-linked immunosorbent assay. Among these specimens, 112 were available for typing: 93 (83.0%) were single-type infections, 9 (8.0%) were mixed-type infections with more than one G or P type, and 10 (8.9%) were G and/or P nontypeable. In contrast to the findings in our last study, from 1996 to 1998, genotype P[4], G2 strains were almost completely absent and P[8], G1 and P[8], G4 strains were dominant, representing more than 80% of the G and P types found. Genotypes G2 and G9 were detected in few samples, and type G3 was completely absent. We identified several uncommon genotype G12 strains, representing the first detections outside of Asia and the United States, by sequencing. Using a genotype G12-specific reverse transcription-PCR, we identified eight (6.7%) positive samples for the 1999 to 2003 period. The high degree of sequence identity between recent G12 isolates from Argentina, the United States, and Asian countries suggests a relatively recent introduction(s) of these strains into humans from a common progenitor. The Argentinean G12 strains belonged to genotype P[9], similar to most of the recently described Asian G12 strains. The finding of G12 strains in several other regions of the world raises the possibility that G12 may be emerging globally and suggests that surveillance for this strain should be conducted routinely.

Detection of group a human rotavirus G9 genotype circulating in Córdoba, Argentina, as early as 1980.

The incidence of human rotavirus G types was determined over a 25-year period (1979-2003) by using reverse transcription-PCR (RT-PCR) to examine 519 stool specimens found to be positive for rotavirus by enzyme linked immunosorbent assay (ELISA) or polyacrylamide gel electrophoresis (PAGE). These stool samples were obtained from children under 3 years old who had been treated for acute diarrhea at public hospitals in Córdoba, Argentina. The present study describes the continued circulation of the common human G types G1 (53.8%), G2 (10.2%), G3 (4.4%), and G4 (27%), and also the detection of the unusual types G8 (0.5%) and G9 (4.2%). Genotype G9 was detected during the 1980-1988 and 1997-2003 periods at relatively low rates. Rotavirus G types distribution was independent of age (1-18 months), gender or out-patient or in-patient status. Unexpectedly, 44.6% of mixed infections were detected, involving common and unusual genotypes. Overall, 95.4% of the typed strains belonged to the most prevalent human serotypes (G1-G4) but the detection of G9 infection throughout this study period highlights the importance of this serotype as a human pathogen.

Characterization of human group C rotavirus in Argentina.

A survey was conducted for identification of human group C rotaviruses in stool specimens taken from children suffering diarrhea in suburban Buenos Aires regions. Among 90 true negative group A samples as defined by ELISA, RT-PCR and PAGE, five were positive by group C specific RT-PCR (VP7 and VP6 genes) and three of these samples exhibited the characteristic 4-3-2-2 dsRNA pattern of group C rotavirus. These results were further confirmed by electron microscopy and by ELISA for detection of group C VP6 specific antigens. Sequence analysis of the VP7 gene from one of these isolates revealed a 97.3-98.6% nucleotide identity and up to 99.1% protein homology with human group C rotavirus strains found scattered throughout the last ten years in other countries. Conversely, similar analysis performed with porcine strains showed a much lower homology degree both at the nucleotide (75.5% nucleotide identity) and amino acid level (85.5% protein homology). Detection of group C rotavirus in children with acute diarrhea in Argentina extends the identification range of this agent in the region and is consistent with previous reported data that demonstrate a global distribution of this virus.

VP7 and VP4 genotyping of human group A rotavirus in Buenos Aires, Argentina.

Specific and sensitive tests for the detection and typing of group A rotavirus strains are needed for a more comprehensive knowledge of the epidemiology of rotaviral infection. In this study 500 stool specimens taken from 1996 to 1998 from children with acute diarrhea in Buenos Aires were examined. Group A rotavirus was unequivocally demonstrated in 62% of the samples tested by enzyme-linked immunosorbent assay (ELISA) for detection of VP6 antigen, polyacrylamide gel electrophoresis of double-stranded RNA, and reverse transcription-PCR (RT-PCR) for amplification of the VP7:G (1, 062 bp) and VP4:P (876 bp) genes. Only five positive specimens were found by RT-PCR but not by ELISA. G and P typing was carried out by nested amplification of variable sequences of the VP7 and the VP4 genes with six G- and five P-type-specific primers (multiplex PCR). Results obtained by this method showed the prevalence of the following G and P types: G1, 39%; G2, 43%; G4, 4%; P[8], 16%; P[4], 71%. Unexpectedly, the G-P type combination most frequently found was G2P[4] (43%) rather than G1P[8] (12%), which is the most commonly found worldwide. Unusual strains of the type G1P[4] accounted for 14% of the total, while mixed infections with more than one type were found in 10% of the samples. Detection of fecal rotavirus-specific immunoglobulin M (IgM) and IgA antibodies in consecutive samples of two patients taken at daily intervals demonstrated that high levels of IgM and IgA antibodies were detected on day 1 after the onset of disease and that the samples remained positive for about 10 days, after which virus shedding was no longer observed. Multiplex PCR offers a sensitive and specific alternative to determine the prevalence of group A rotavirus G and P types and to identify the emergence of uncommon strains, whereas detection of fecal IgM and IgA antibodies represents a useful supplement to virus detection for the diagnosis of current or recently acquired infections.

Hepatitis B virus antigens in peripheral blood mononuclear cells during the course of viral infection.

We studied the expression of surface (HBsAg) and core (HBcAg) proteins of hepatitis B virus (HBV) on the surface of peripheral blood mononuclear cells (PBMC) from HBV-infected patients. A total of 122 patients with different liver viral diseases was analyzed by indirect immunofluorescence with monoclonal antibodies. The 35 patients with HBV chronic active hepatitis (CAH) and 38 of 60 patients with acute hepatitis B (63%) expressed HBsAg on the PBMC. No expression was detected on the cells from both normal and HBV-unrelated viral hepatitis control groups. Serial follow-up of patients with acute hepatitis B showed that HBsAg expression by PBMC tended to be undetectable 4 months after the onset of the disease and at the same time the clinical improvement was evident. Cell cultures of EBV-transformed B lymphocytes were established from PBMC of HBV-infected patients; immunoelectron microscopy demonstrated the HBsAg on the cellular membrane. One-third of HBV-infected patients who were studied showed the expression of HBcAg by PBMC. HBcAg was detected in patients with acute hepatitis B at the early stage of infection. The cells of these patients also expressed HBsAg in PBMC. In CAH patients, a positive association was observed between the expression of HBcAg and the presence of serum HBeAg.