ABSTRACT
KEY MESSAGE: An efficient and improved transformation method for functional genetics studies in S. italica, being a boon for the Setaria scientific community and for crop improvement. Foxtail millet (Setaria italica) is a short life cycle C4 plant, with sequenced genome, and a potential model plant for C4 species. S. italica is also important on a global food security and healthiness context due to its importance in arid and semi-arid areas. However, despite its importance, there are just few transformation protocols directed to this species. The current protocols reached about 5.5-9% of efficiency, which do not make it a valuable model organism. Different types of explants were used in the above mentioned methods, such as immature and mature inflorescence and shoot apex. However, these techniques have many limitations, such as unavailability of explants throughout the year and a crucial, laborious and considerable time-consuming selection. Aiming a simplified and efficient methodology, we adopted dry mature seeds as explants, which are available in abundance, are constant along the year and well responsive to tissue culture, in addition to a differentiated approach that reaches on an average 19.2% transformation efficiency of S. italica. Thus, we propose a protocol that optimizes the transformation efficiency of this cereal crop allowing a high increase on transformation and regeneration rates. Our transformation protocol provides an interesting tool for Setaria community research as well as enables new strategies for breeding enhanced productivity in the species.
Subject(s)
Regeneration/genetics , Setaria Plant/genetics , Transformation, Genetic , Agrobacterium tumefaciens/genetics , Cell Culture Techniques/methods , Cells, Cultured , Edible Grain/genetics , Edible Grain/metabolism , Genetic Techniques , Genetic Vectors , Phenotype , Plant Breeding , Plant Leaves/genetics , Plant Leaves/growth & development , Plant Leaves/metabolism , Plants, Genetically Modified , RNA Interference , Regeneration/physiology , Seeds/drug effects , Seeds/genetics , Setaria Plant/metabolism , Setaria Plant/microbiology , Setaria Plant/physiologyABSTRACT
With the objective of characterizing Canine parvovirus (CPV) from some suspected fecal samples of dogs collected from the Veterinarian Hospital in Belém city, five positive samples were found by PCR assay and an update molecular characterization was provided of the CPV-2 circulation in Belém. Through sequencing of the complete DNA sequences (NS1, NS2, VP1, and VP2 genes), the CPV-2 strain was identified as CPV-2b (Asn426Asp) circulating in Belém. The CPV-2b strain with a different change at the position Tyr324Leu was detected in all samples assessed and thus reported for the first time for the scientific community. Phylogenetic analysis indicated that Belém CPV-2b and CPV-2a strains would be related to a cluster with samples after the 1990s, suggesting that CPV-2b in Belém originated from CPV-2a circulating in Brazil after the 1990s. Potential recombination events were analyzed using RDP4 and SplitsTree4; therefore, results suggest that CPV-2 sequences here described were not potentially recombination events. Continuous monitoring and molecular characterization of CPV-2 samples are needed not only to identify possible genetic and antigenic changes that may interfere with the effectiveness of vaccines but also to bring a better understanding of the mechanisms that drive the evolution of CPV-2 in Brazil.
Subject(s)
Genome, Viral , Parvovirus, Canine/genetics , Polymorphism, Genetic , Animals , Brazil , Dogs/virology , Feces/virology , Parvoviridae Infections/veterinary , Parvoviridae Infections/virology , Parvovirus, Canine/classification , Parvovirus, Canine/isolation & purification , Phylogeny , Recombination, GeneticABSTRACT
Sexually transmitted infections are an important cause of morbidity among sexually active women worldwide, and have been implicated as cofactors in the pathogenesis of cervical cancer. We investigated the prevalence of human papillomavirus (HPV), Chlamydia trachomatis (CT), and Trichomonas vaginalis (TV), and accessed the diversity of HPV in women with normal and abnormal cytology in Manaus, Brazil. We used polymerase chain reaction and HPV genotyping by direct sequencing. The chi-square test was used to calculate the absolute and relative frequencies of the categorical variables, and Fisher's test was used when P < 0.05. The level of significance was set at 5%. All statistical analyses were performed using R 2.9.0. There were statistically significant differences in age (P = 0.0395), education level (P = 0.0131), sexual partners (P = 0.0211), condom use (P = 0.0039), marital status (P < 0.0001), and pregnancy (P = 0.0003) between the normal and abnormal groups. HPV DNA was found in 36.56 and 93.88% of subjects in the normal and abnormal groups, respectively. A total of 19 genotypes were detected; HPV16 was the most common, followed by HPV58. The percentages of TV and CT DNA were 18.04 and 9.02% in the normal group, respectively. The percentages of HPV/TV and HPV/CT coinfection were 12.5% each in women with normal cytology. These findings improve our understanding of HPV, CT, and TV, and the distribution of HPV types, which may be relevant to vaccination strategies for protecting women from the north of Brazil from cervical cancers and precancerous lesions.
Subject(s)
Chlamydia Infections/epidemiology , Chlamydia trachomatis/genetics , Genotype , Papillomaviridae/genetics , Papillomavirus Infections/epidemiology , Trichomonas Vaginitis/epidemiology , Trichomonas vaginalis/genetics , Adolescent , Adult , Brazil , Chlamydia Infections/microbiology , Chlamydia Infections/pathology , Chlamydia trachomatis/isolation & purification , Female , Humans , Middle Aged , Papillomaviridae/isolation & purification , Papillomavirus Infections/pathology , Papillomavirus Infections/virology , Prevalence , Trichomonas Vaginitis/parasitology , Trichomonas Vaginitis/pathology , Trichomonas vaginalis/isolation & purificationABSTRACT
Sapindus saponaria L. of Sapindaceae family is popularly known as soldier soap and is found in Central and South America. A study of such medicinal plants might reveal a more complex diversity of microorganisms as compared to non-medicinal plants, considering their metabolic potential and the chemical communication between their natural microbiota. Rhizosphere is a highly diverse microbial habitat with respect to both the diversity of species and the size of the community. Rhizosphere bacteriome associated with medicinal plant S. saponaria is still poorly known. The objective of this study was to assess the rhizosphere microbiome of the medicinal plant S. saponaria using pyrosequencing, a culture-independent approach that is increasingly being used to estimate the number of bacterial species present in different environments. In their rhizosphere microbiome, 26 phyla were identified from 5089 sequences of 16S rRNA gene, with a predominance of Actinobacteria (33.54%), Acidobacteria (22.62%), and Proteobacteria (24.72%). The rarefaction curve showed a linear increase, with 2660 operational taxonomic units at 3% distance sequence dissimilarity, indicating that the rhizosphere microbiome associated with S. saponaria was highly diverse with groups of bacteria important for soil management, which could be further exploited for agricultural and biotechnological purposes.
Subject(s)
Microbiota , Plants, Medicinal/microbiology , Rhizosphere , Sapindus/genetics , Sapindus/microbiology , Sequence Analysis, DNA/methods , Temperature , Bacteria/classification , Biodiversity , Phylogeny , Plants, Medicinal/genetics , RNA, Ribosomal, 16S/genetics , Soil MicrobiologyABSTRACT
BACKGROUND: Endoscopic submucosal dissection (ESD) of extensive superficial cancers of the esophagus may progress with high rates of postoperative stenosis, resulting in significantly decreased quality of life. Several therapies are performed to prevent this, but have not yet been compared in a systematic review. METHODS: A systematic review of the literature and meta-analysis were performed using the MEDLINE, Embase, Cochrane, LILACS, Scopus, and CINAHL databases. Clinical trials and observational studies were searched from March 2014 to February 2015. Search terms included: endoscopy, ESD, esophageal stenosis, and esophageal stricture. Three retrospective and four prospective (three randomized) cohort studies were selected and involved 249 patients with superficial esophageal neoplasia who underwent ESD, at least two-thirds of the circumference. We grouped trials comparing different techniques to prevent esophagus stenosis post-ESD. RESULTS: We conducted different meta-analyses on randomized clinical trials (RCT), non-RCT, and global analysis. In RCT (three studies, n = 85), the preventive therapy decreased the risk of stenosis (risk difference = -0.36, 95 % CI -0.55 to -0.18, P = 0.0001). Two studies (one randomized and one non-randomized, n = 55) showed that preventative therapy lowered the average number of endoscopy dilatations (mean difference = -8.57, 95 % CI -13.88 to -3.25, P < 0.002). There were no significant differences in the three RCT studies (n = 85) in complication rates between patients with preventative therapy and those without (risk difference = 0.02, 95 % CI -0.09 to 0.14, P = 0.68). CONCLUSIONS: The use of preventive therapy after extensive ESD of the esophagus reduces the risk of stenosis and the number of endoscopic dilatations for resolution of stenosis without increasing the number of complications.
Subject(s)
Adenocarcinoma/surgery , Carcinoma, Squamous Cell/surgery , Endoscopic Mucosal Resection/methods , Esophageal Neoplasms/surgery , Esophageal Stenosis/prevention & control , Esophagoscopy/methods , Postoperative Complications/prevention & control , Endoscopic Mucosal Resection/adverse effects , Esophageal Stenosis/etiology , Esophagoscopy/adverse effects , Humans , Postoperative Complications/etiology , Quality of LifeABSTRACT
CONTEXT: No long-term studies have compared centrally acting drugs for treating obesity. OBJECTIVE: To compare the efficacy and safety of diethylpropion (DEP), fenproporex (FEN), mazindol (MZD), fluoxetine (FXT) and sibutramine (SIB) in promoting weight loss. DESIGN AND SETTING: A prospective, randomized, placebo (PCB)-controlled study conducted at a single academic institution. PATIENTS: A total of 174 obese premenopausal women. INTERVENTION: Participants randomly received DEP 75 mg (n=28), FEN 25 mg (n=29), MZD 2 mg (n=29), SIB 15 mg (n=30), FXT 20 mg (n=29) or PCB (n=29) daily over 52 weeks. Diet and physical activity were encouraged. MAIN OUTCOME MEASURES: The primary endpoints were changes in body weight and the proportion of women who achieved at least 5% weight loss by week 52 in the intent-to-treat population. Other measurements included anthropometry, safety, metabolic and cardiovascular parameters. RESULTS: Weight loss was greater than PCB (-3.1±4.3 kg) with DEP (-10.0±6.4 kg; P<0.001), SIB (-9.5±5.9 kg; P<0.001), FEN (-7.8±6.9 kg; P<0.01) and MZD (-7.4±4.9 kg; P<0.01) but not with FXT (-2.5±4.1 kg). Ten (33.3%) women lost⩾5% of their initial weight with PCB, compared with 20 (71.4%; P<0.001) with DEP, 20 (69%; P<0.02) with FEN, 21 (72.4%; P<0.01) with MZD, 22 (73.3%; P<0.001) with SIB and 10 (35.5%) with FXT. Each medically treated group experienced more adverse events compared with PCB (P<0.001). Compared with PCB, constipation was more prevalent with DEP, SIB and MZD (P<0.01); anxiety was more prevalent with DEP (P=0.01); and irritability occurred more frequently with DEP and FEN (P=0.02). Significant improvements in the depression and anxiety scores, binge-eating episodes and quality of life correlated with weight loss. CONCLUSION: The centrally acting drugs DEP, FEN, MZD and SIB were more effective than PCB in promoting weight loss in obese premenopausal women, with a satisfactory benefit-risk profile.
Subject(s)
Amphetamines/therapeutic use , Anti-Obesity Agents/therapeutic use , Cyclobutanes/therapeutic use , Diethylpropion/therapeutic use , Fluoxetine/therapeutic use , Mazindol/therapeutic use , Obesity/drug therapy , Weight Loss/drug effects , Adult , Body Mass Index , Brazil , Diet, Reducing , Female , Follow-Up Studies , Humans , Obesity/prevention & control , Prospective Studies , Surveys and Questionnaires , Treatment OutcomeABSTRACT
Thirty-four microsatellite markers (SSRs) were identified in EST and BAC clones from Musa acuminata burmannicoides var. Calcutta 4 and validated in 22 Musa genotypes from the Banana Germplasm Bank of Embrapa-CNPMF, which includes wild and improved diploids. The number of alleles per locus ranged from 2 to 14. The markers were considered highly informative based on their polymorphism information content values; more than 50% were above 0.5. These SSRs will be useful for banana breeding programs, for studies of genetic diversity, germplasm characterization and selection, development of saturated genetic linkage maps, and marker assisted selection.
Subject(s)
Genetic Markers , Microsatellite Repeats/genetics , Musa/genetics , Base Sequence , DNA Primers , DNA, Plant/genetics , Polymerase Chain ReactionABSTRACT
Superparamagnetic iron-oxide (SPIO) particles were synthesized by the co-precipitation method and the oleic acid-coated SPIO (OA-SPIO) was then obtained by a surface grafting procedure. A stock sample of magnetic oil (MO) with 1.6% particle volume fraction (VF) was obtained by dispersing the OA-SPIO in insulating naphthenic oil. The MO stock sample was diluted in the same naphthenic oil to yield MO with 0.1, 0.04, 0.02, and 0.01% VF. Moreover, the 0.04% VF MO sample was manipulated to yield MO samples with water content of 26, 37, and 63 mg L(-1). The spinel structure of OA-SPIO was assessed by XRD and the average diameter of 8.3 nm was provided by TEM analysis. The saturation magnetization at room temperature (RT) was 70 emu/g and no remanence or coercivity was observed. The average hydrodynamic diameter (D(H)) of the colloidal particles suspended within the 0.04% VF MO sample was 58 nm. After aging for 30 days at RT no change was observed for the lowest water content MO sample (26 mg L(-1)). However, D(H) equals to 270 nm was observed for the highest water content MO sample (63 mg L(-1)). The MO samples with 26 mg L(-1) water content were found stable under heating at 90 degrees C for all VF investigated. We found the insulation resistance dropping significantly as VF and temperature increases. The lowest value found was 11 GOhms for the 0.1% VF at 60 degrees C, which is an acceptable value for MO.
ABSTRACT
A recombinant bovine herpesvirus 5 lacking thymidine kinase and glycoprotein E genes (BoHV-5gEΔTKΔ) was evaluated as a live experimental vaccine. In a first experiment, ten-months-old calves were vaccinated intramuscularly (n=9) or remained as controls (n=8) and 42 days later were challenged with BoHV-5 or BoHV-1 intranasally. The four control calves challenged with BoHV-5 developed severe depression and neurological signs and were euthanized in extremis at days 13 and 14 pos-infection (pi); the five vaccinated animals challenged with BoHV-5 remained healthy. The titers of virus shedding were reduced (p<0.01) from days 3 to 7 post-infection (pi) in vaccinated animals. Control and vaccinated calves challenged with BoHV-1 presented mild transient respiratory signs; yet the magnitude of virus shedding was reduced (p<0.05) in vaccinated animals (days 5, 9 and 11pi). In a second experiment, young calves (100-120 days-old) were vaccinated (n=15) or kept as controls (n=5) and subsequently challenged with a BoHV-1 isolate. Control calves developed moderate to severe rhinitis and respiratory distress; two were euthanized in extremis at days 5 and 9 pi, respectively. In contrast, vaccinated animals were protected from challenge and only a few developed mild and transient nasal signs. The duration and titers of virus shedding after challenge were reduced (p<0.05) in vaccinated animals comparing to controls. In both experiments, vaccinated animals developed antibodies to gE only after challenge. These results demonstrate homologous and heterologous protection and are promising towards the use of the recombinant BoHV-5gEΔTKΔ in vaccine formulations to control BoHV-5 and BoHV-1 infections.
Subject(s)
Herpesvirus 1, Bovine/pathogenicity , Herpesvirus 5, Bovine/immunology , Infectious Bovine Rhinotracheitis/prevention & control , Thymidine Kinase/genetics , Viral Envelope Proteins/genetics , Viral Vaccines/immunology , Animals , Antibodies, Viral/blood , Antibody Formation , Cattle , Female , Herpesvirus 1, Bovine/immunology , Herpesvirus 5, Bovine/enzymology , Herpesvirus 5, Bovine/genetics , Infectious Bovine Rhinotracheitis/immunology , Male , Vaccines, Attenuated/immunology , Virus SheddingABSTRACT
This article describes an investigation on the virulence/attenuation of bovine herpesvirus type 5 (BoHV-5) recombinants deleted in the genes encoding glycoprotein E (BoHV-5gEΔ), thymidine kinase (BoHV-5TKΔ), and both gE and TK (BoHV-5gEΔTKΔ). Seronegative calves (80 to 90 days-old) inoculated with the parental strain (SV-507/99, n=5) shed virus in nasal secretions for up to 15 days (average 10.8 days). Duration of virus shedding was 11 days for BoHV-5gΔ, 9.6 days for BoHV-5TKΔ and 6.2 days for BoHV-5gEΔTKΔ groups. The highest titers were observed between days 1 and 6 post-infection (pi) for SV-507/99 (10(6.8)TCID50/mL), 10(5.1)TCID50/mL (BoHV-5gEΔ), 10(5.9)TCID50/mL (BoHV-5TKΔ) and 10(4.7)TCID50/mL (BoHV-5gEΔTKΔ). Calves inoculated with the parental virus presented anorexia, profound apathy and loss of body condition. Two calves were euthanized in extremis on days 10 and 11 pi; infectious virus was recovered from several areas of the brain. In contrast, calves inoculated with the recombinants remained healthy and a few presented a mild and transient nasal secretion. Dexamethasone (Dx) administration at day 42 pi resulted in virus shedding by all controls calves (mean duration 3.7 days), by 2/5 of BoHV-5TKΔ calves (two days) and 2/5 of BoHV-5gEΔ (one day). No virus shedding was detected in BoHV-5gEΔTKΔ calves upon Dx treatment. PCR examination of brain sections of calves euthanized at day 30 post Dx treatment revealed the presence of latent viral DNA widely distributed in the brain of SV-507/99 calves. Latent viral DNA was detected in a few sections (3/30) of the brains of BoHV-5gEΔ calves and was not detected in the brains of calves inoculated with BoHV-5TKΔ and BoHV-5gEΔTKΔ. These results show that the single BoHV-5 mutants (gE and tk-deleted) are attenuated for calves and establish and/or reactivate latent infection inefficiently. The double mutant BoHV-5gEΔTKΔ is fully attenuated and appears not to establish or not reactivate efficiently from latent infection. Thus, these recombinants, especially the double mutant BoHV-5gEΔTKΔ, display an adequate phenotype for use in modified-live vaccine formulations.(AU)
Este artigo descreve uma investigação da virulência/atenuação de recombinantes do herpesvírus bovino tipo 5 (BoHV-5) com deleções nos genes da glicoproteína E (BoHV-5gEΔ), timidina quinase (BoHV-5TKΔ), e ambos gE e TK (BoHV-5gEΔTKΔ). Bezerros soronegativos (80-90 dias de idade) inoculados com o vírus parental SV-507/99 (n=5) excretaram o vírus em secreções nasais por até 15 dias (média de 10,8 dias). Nos animais inoculados com os recombinantes, a duração da excreção viral foi de 11 dias (BoHV-5gEΔ), 9,6 dias (BoHV-5TKΔ) e 6,2 dias (BoHV-5gEΔTKΔ). Os maiores títulos foram observados entre os dias 1 e 6 pós-inoculação (pi), sendo de 10(6,8)TCID50/mL para o SV-507/99, 10(5,1)TCID50/mL (BoHV-5gEΔ), 10(5,9)TCID50/mL (BoHV-5TKΔ) e 10(4,7)TCIΔ50/mL (BoHV-5gEΔTKΔ). Os bezerros inoculados com o vírus parental apresentaram anorexia e apatia; três deles mostraram apatia profunda e perda da condição corporal. Dois bezerros foram eutanasiados in extremis nos dias 10 e 11 pi, respectivamente e o vírus foi isolado de várias regiões do encéfalo. Já os bezerros inoculados com os recombinantes permaneceram saudáveis; alguns apresentaram uma secreção nasal serosa transitória. Administração de dexametasona (Dx) no dia 42 pi resultou em excreção viral por todos os bezerros inoculados com o vírus parental (duração média de 3,7 dias), por 2 de 5 bezerros dos grupos BoHV-5TKΔ (dois dias) e BoHV-5gEΔ (um dia). Os bezerros inoculados com o duplo mutante BoHV-5gEΔTKΔ não excretaram o vírus após o tratamento com Dx. Pesquisa de DNA viral por PCR no dia 30 pós-Dx revelou uma ampla distribuição do DNA do vírus parental no encéfalo; poucas seções (3/30) foram positivas no encéfalo dos animais do grupo BoHV-5gEΔ, e não detectou-se DNA latente no encéfalo dos animais dos grupos BoHV-5TKΔ e BoHV-5gEΔTKΔ. Esses resultados demonstram que os mutantes simples (gE and tk-deletados) são atenuados para bezerros e estabelecem e/ou reativam infecção latente ineficientemente. Já o duplo mutante BoHV-5gEΔTKΔ é atenuado e parece não estabelecer e/ou não reativar eficientemente a infecção latente. Portanto, os vírus recombinantes, e em especial o duplo mutante BoHV-5gEΔTKΔ apresentam um fenótipo compatível com a sua inclusão em vacinas vivas modificadas.(AU)
Subject(s)
Animals , Herpesvirus 5, Bovine/pathogenicity , Glycoproteins/adverse effects , Glycoproteins , Thymidine Kinase , Recombinant ProteinsABSTRACT
This article describes an investigation on the virulence/attenuation of bovine herpesvirus type 5 (BoHV-5) recombinants deleted in the genes encoding glycoprotein E (BoHV-5gEΔ), thymidine kinase (BoHV-5TKΔ), and both gE and TK (BoHV-5gEΔTKΔ). Seronegative calves (80 to 90 days-old) inoculated with the parental strain (SV-507/99, n=5) shed virus in nasal secretions for up to 15 days (average 10.8 days). Duration of virus shedding was 11 days for BoHV-5gΔ, 9.6 days for BoHV-5TKΔ and 6.2 days for BoHV-5gEΔTKΔ groups. The highest titers were observed between days 1 and 6 post-infection (pi) for SV-507/99 (10(6.8)TCID50/mL), 10(5.1)TCID50/mL (BoHV-5gEΔ), 10(5.9)TCID50/mL (BoHV-5TKΔ) and 10(4.7)TCID50/mL (BoHV-5gEΔTKΔ). Calves inoculated with the parental virus presented anorexia, profound apathy and loss of body condition. Two calves were euthanized in extremis on days 10 and 11 pi; infectious virus was recovered from several areas of the brain. In contrast, calves inoculated with the recombinants remained healthy and a few presented a mild and transient nasal secretion. Dexamethasone (Dx) administration at day 42 pi resulted in virus shedding by all controls calves (mean duration 3.7 days), by 2/5 of BoHV-5TKΔ calves (two days) and 2/5 of BoHV-5gEΔ (one day). No virus shedding was detected in BoHV-5gEΔTKΔ calves upon Dx treatment. PCR examination of brain sections of calves euthanized at day 30 post Dx treatment revealed the presence of latent viral DNA widely distributed in the brain of SV-507/99 calves. Latent viral DNA was detected in a few sections (3/30) of the brains of BoHV-5gEΔ calves and was not detected in the brains of calves inoculated with BoHV-5TKΔ and BoHV-5gEΔTKΔ. These results show that the single BoHV-5 mutants (gE and tk-deleted) are attenuated for calves and establish and/or reactivate latent infection inefficiently. The double mutant BoHV-5gEΔTKΔ is fully attenuated and appears not to establish or not reactivate efficiently from latent infection. Thus, these recombinants, especially the double mutant BoHV-5gEΔTKΔ, display an adequate phenotype for use in modified-live vaccine formulations.
Este artigo descreve uma investigação da virulência/atenuação de recombinantes do herpesvírus bovino tipo 5 (BoHV-5) com deleções nos genes da glicoproteína E (BoHV-5gEΔ), timidina quinase (BoHV-5TKΔ), e ambos gE e TK (BoHV-5gEΔTKΔ). Bezerros soronegativos (80-90 dias de idade) inoculados com o vírus parental SV-507/99 (n=5) excretaram o vírus em secreções nasais por até 15 dias (média de 10,8 dias). Nos animais inoculados com os recombinantes, a duração da excreção viral foi de 11 dias (BoHV-5gEΔ), 9,6 dias (BoHV-5TKΔ) e 6,2 dias (BoHV-5gEΔTKΔ). Os maiores títulos foram observados entre os dias 1 e 6 pós-inoculação (pi), sendo de 10(6,8)TCID50/mL para o SV-507/99, 10(5,1)TCID50/mL (BoHV-5gEΔ), 10(5,9)TCID50/mL (BoHV-5TKΔ) e 10(4,7)TCIΔ50/mL (BoHV-5gEΔTKΔ). Os bezerros inoculados com o vírus parental apresentaram anorexia e apatia; três deles mostraram apatia profunda e perda da condição corporal. Dois bezerros foram eutanasiados in extremis nos dias 10 e 11 pi, respectivamente e o vírus foi isolado de várias regiões do encéfalo. Já os bezerros inoculados com os recombinantes permaneceram saudáveis; alguns apresentaram uma secreção nasal serosa transitória. Administração de dexametasona (Dx) no dia 42 pi resultou em excreção viral por todos os bezerros inoculados com o vírus parental (duração média de 3,7 dias), por 2 de 5 bezerros dos grupos BoHV-5TKΔ (dois dias) e BoHV-5gEΔ (um dia). Os bezerros inoculados com o duplo mutante BoHV-5gEΔTKΔ não excretaram o vírus após o tratamento com Dx. Pesquisa de DNA viral por PCR no dia 30 pós-Dx revelou uma ampla distribuição do DNA do vírus parental no encéfalo; poucas seções (3/30) foram positivas no encéfalo dos animais do grupo BoHV-5gEΔ, e não detectou-se DNA latente no encéfalo dos animais dos grupos BoHV-5TKΔ e BoHV-5gEΔTKΔ. Esses resultados demonstram que os mutantes simples (gE and tk-deletados) são atenuados para bezerros e estabelecem e/ou reativam infecção latente ineficientemente. Já o duplo mutante BoHV-5gEΔTKΔ é atenuado e parece não estabelecer e/ou não reativar eficientemente a infecção latente. Portanto, os vírus recombinantes, e em especial o duplo mutante BoHV-5gEΔTKΔ apresentam um fenótipo compatível com a sua inclusão em vacinas vivas modificadas.
Subject(s)
Animals , Glycoproteins/adverse effects , Glycoproteins , /pathogenicity , Thymidine Kinase , Recombinant ProteinsABSTRACT
Infection by human papillomavirus (HPV) is one of the primary causes of mortality by cancer in northern Brazil. Sexually active women from Manaus, Amazonas, without cytological alterations and women with pre-malignant and malignant cytological alterations were examined for HPV virus, identified via PCR and sequencing. The target region for this study was part of the L1 capsid gene of HPV. Twenty-three samples that were PCR-positive were sequenced. Analysis of 336 bp demonstrated a high incidence of high-risk HPV types in the population of Manaus, identified as HPVs 16, 33, 58, 66, 68. HPV type 16 was the most prevalent, presenting two variants similar to the Asian-American (AA) and East-Asian type (As) variants. A rare HPV type 13 related to "Heck's disease" was also detected. This preliminary provides important information about the HPV circulating in Amazonas State.
Subject(s)
DNA, Viral/genetics , Human papillomavirus 16/genetics , Brazil , Capsid Proteins/genetics , Female , Human papillomavirus 16/classification , Human papillomavirus 16/pathogenicity , Humans , Polymerase Chain Reaction , Sequence Analysis, DNA , Uterine Cervical Neoplasms/virologyABSTRACT
This research evaluated the effects of Brånemark protocol on electromyography of the masseter and temporal muscles. The patients were divided into two groups: Group I: patients who wore an implant-supported prosthesis in the mandibular arch following Brånemark protocol, and maxillary removable complete dentures; Group II: dentate individuals (control). Electromyography was carried out at rest, right (RL) and left (LL) laterality, protrusion and maximum voluntary contraction (MVC). Data were compared by t-test. At rest, a higher electromyographic (EMG) activity was observed in Group I, and the values were significant in the right masseter and left temporal muscles. In RL, there were statistically significant differences for right masseter (P < 0·01), left masseter and temporal muscles and for the left temporal muscle in LL (P < 0·05). In protrusion, Group I presented a higher EMG activity, and there was a statistically significant difference for the right masseter muscle (RM) (P < 0·05). In MVC, the EMG values were higher in Group II (control), but significant just for the right temporal muscle (P < 0·05). In conclusion, individuals with mandibular fixed dentures supported according to the Brånemark protocol and maxillary removable complete dentures showed a higher activity of masticatory muscles during the mandibular postural clinical conditions examined; however, in the MVC, Group I presented lower values when compared to dentate group. Despite presenting different EMG values in all of the clinical conditions, both groups showed similar EMG patterns of muscle activation which demonstrates that the proposed treatment can be considered as a good option for oral rehabilitation.
Subject(s)
Dental Prosthesis, Implant-Supported , Denture, Complete, Upper , Masseter Muscle/physiology , Temporal Muscle/physiology , Aged , Aged, 80 and over , Brazil , Case-Control Studies , Electromyography/methods , Female , Humans , Jaw Relation Record , Male , Mastication/physiology , Middle Aged , Muscle Contraction/physiology , Practice Guidelines as TopicABSTRACT
Records of the gastropod Melanoides tuberculatus (Müller, 1774), family Thiaridae, in the Piranhas-Assu River basin in Rio Grande do Norte reveal the dispersal of this native Southeast Asian and East African species into aquatic environments of the Brazilian semiarid region, including artificial environments (reservoirs) and lotic systems. The eutrophic conditions of the local waterbodies appear to favor the present situation, where this invasive species reaches extremely high densities, sometimes over 10,000 ind x m(-2) as in Armando Ribeiro Gonçalves Reservoir. These observations indicate the immediate need for new studies on the spatial distribution of the species and its potential impact on the biodiversity and water quality of the waterbodies of the semiarid region of the state. Implantation of regular and systematic monitoring of the aquatic resources of the region is urgently required.
Subject(s)
Ecosystem , Environmental Monitoring , Snails/physiology , Animals , Brazil , Population Density , Population Dynamics , Rivers , Snails/classificationABSTRACT
The objective of this study was to provide information on the histological characteristics of the gonads of male and female Armases rubripes crabs, and to try to establish a relationship between the microscopic and macroscopic stages previously identified. Thirty-six crabs were collected by hand between February 2003 and January 2004 in banks of Spartina alterniflora on Sahy Beach in Mangaratiba, Rio de Janeiro state, Brazil. The histological analysis of the ovaries of A. rubripes demonstrated a gradual process of development of the oocytes. According to their cellular characteristics, five types of cells were distinguished: oogonia, oocyte I, oocyte II, oocyte III and oocyte IV. The ovaries showed four stages during gonadal activity: stage I (rudimentary), stage II (developing or maturing), stage III (developed or mature) and stage IV (resting). The results of the histochemical analyses showed that the ovaries vary according to the gonad development stage. The histological aspect of one section of the male gonad was always the same in all of the seminiferous tubules, where the lumen of these tubules always contained spermatozoa and/or spermatids. It was not possible to characterize the three stages of gonad development in the males. This agrees with previous reports in the literature. However, in the females there was a relationship between the gonad stages distinguished macroscopically and the results obtained through the histological and histochemical analysis, due to the presence of different cell types, as well as the lysis process and reabsorption of the oocytes in spent females.
Subject(s)
Brachyura/growth & development , Oocytes/growth & development , Ovary/growth & development , Sexual Maturation/physiology , Testis/growth & development , Animals , Brachyura/chemistry , Brachyura/cytology , Female , Histocytochemistry , Male , Oocytes/cytology , Ovary/chemistry , Ovary/cytology , Testis/chemistry , Testis/cytologyABSTRACT
The objective of this study was to provide information on the histological characteristics of the gonads of male and female Armases rubripes crabs, and to try to establish a relationship between the microscopic and macroscopic stages previously identified. Thirty-six crabs were collected by hand between February 2003 and January 2004 in banks of Spartina alterniflora on Sahy Beach in Mangaratiba, Rio de Janeiro state, Brazil. The histological analysis of the ovaries of A. rubripes demonstrated a gradual process of development of the oocytes. According to their cellular characteristics, five types of cells were distinguished: oogonia, oocyte I, oocyte II, oocyte III and oocyte IV. The ovaries showed four stages during gonadal activity: stage I (rudimentary), stage II (developing or maturing), stage III (developed or mature) and stage IV (resting). The results of the histochemical analyses showed that the ovaries vary according to the gonad development stage. The histological aspect of one section of the male gonad was always the same in all of the seminiferous tubules, where the lumen of these tubules always contained spermatozoa and/or spermatids. It was not possible to characterize the three stages of gonad development in the males. This agrees with previous reports in the literature. However, in the females there was a relationship between the gonad stages distinguished macroscopically and the results obtained through the histological and histochemical analysis, due to the presence of different cell types, as well as the lysis process and reabsorption of the oocytes in spent females.
O objetivo deste estudo foi fornecer informações sobre as características histológicas das gônadas de machos e de fêmeas de Armases rubripes, tentando estabelecer uma relação entre os estágios microscópicos e os macroscópicos anteriormente identificados. Foram coletados manualmente 36 caranguejos, durante o período de fevereiro de 2003 a janeiro de 2004, em bancos de Spartina alterniflora na praia do Sahy Mangaratiba, Estado do Rio de Janeiro. A análise histológica dos ovários de A. rubripes demonstrou um processo gradual de desenvolvimento dos oócitos. De acordo com sua característica celular, cinco tipos de células foram distinguidos: ovogônias, oócito I, oócito II, oócito III, oócito IV. Os ovários revelaram quatro estágios de atividade gonadal: estágio I (rudimentar), estágio II (em desenvolvimento ou em maturação), estágio III (desenvolvido ou maduro), estágio IV (desovada). Os resultados das análises histoquímicas permitem afirmar que os ovários variam de acordo com o estágio de desenvolvimento gonadal. O aspecto histológico de uma sessão de gônada masculina é sempre o mesmo em todos os túbulos seminíferos, onde o lúmen deste túbulo sempre contém espermatozóides e/ou espermátides. Não foi possível a caracterização de três estágios de desenvolvimento gonadal em machos, conforme descrito previamente na literatura. Entretanto, em fêmeas, houve uma relação entre os estágios gonadais distinguidos macroscopicamente e os resultados obtidos através da análise histológica e histoquímica, devido à presença de diferentes tipos celulares, assim como processo de lise e reabsorção dos oócitos em fêmeas desovadas.
Subject(s)
Animals , Female , Male , Brachyura/growth & development , Oocytes/growth & development , Ovary/growth & development , Sexual Maturation/physiology , Testis/growth & development , Brachyura/chemistry , Brachyura/cytology , Histocytochemistry , Oocytes/cytology , Ovary/chemistry , Ovary/cytology , Testis/chemistry , Testis/cytologyABSTRACT
The regional distribution and relative frequency of endocrine cells in the stomach and intestine of Phyllostomidae: Lonchorhina aurita and Molossidae: Molossus molossus bats were studied immunohistochemically. Three types of immunoreactive (IR) endocrine cells--to serotonin (5-HT), gastrin (GAS) and enteroglucagon (GLUC)--were found in the gastric mucosa and four types of IR cells were identified in the intestinal mucosa. This study showed an interespecfic difference in the regional distribution and relative frequency of endocrine cells in the Chiropteran alimentary tract.
Subject(s)
Chiroptera , Enteroendocrine Cells/cytology , Gastric Mucosa/cytology , Intestinal Mucosa/cytology , Animals , Cell Count , Enteroendocrine Cells/immunology , Female , Immune Sera/immunology , Immunohistochemistry/veterinary , MaleABSTRACT
The regional distribution and relative frequency of endocrine cells in the stomach and intestine of Phyllostomidae: Lonchorhina aurita and Molossidae: Molossus molossus bats were studied immunohistochemically. Three types of immunoreactive (IR) endocrine cells - to serotonin (5-HT), gastrin (GAS) and enteroglucagon (GLUC) - were found in the gastric mucosa and four types of IR cells were identified in the intestinal mucosa. This study showed an interespecfic difference in the regional distribution and relative frequency of endocrine cells in the Chiropteran alimentary tract.
A distribuição regional e a freqüência relativa das células endócrinas no estômago e intestino dos morcegos insetívoros Phyllostomidae: Lonchorhina aurita e Mormoopidae: Molossus molossus foram estudadas pelo método de imunohistoquímica. Três tipos de células endócrinas imunorreativas (IR) à serotonina (5-HT), gastrina (GAS) e enteroglucagon (GLUC) foram localizadas na mucosa gástrica e quatro tipos de células endócrinas IR à 5-HT, GAS, colecistoquinina (CCK) e GLUC foram identificadas na mucosa intestinal. Este estudo mostrou uma diferença interespecífica na distribuição regional e na freqüência relativa das células endócrinas no trato alimentar de Chiropteros.
Subject(s)
Animals , Female , Male , Chiroptera , Enteroendocrine Cells/cytology , Gastric Mucosa/cytology , Intestinal Mucosa/cytology , Cell Count , Enteroendocrine Cells/immunology , Immune Sera/immunology , Immunohistochemistry/veterinaryABSTRACT
Measurements of ammonium and phosphate excretion by the Chironomus larvae were conducted in order to evaluate the importance of these chironomids for the internal loads of a small eutrophic urban reservoir. Ammonium and phosphate excretion rates by Chironomus larvae of small size (6-10 mm total length) were significantly higher than those of the Chironomids having medium (9-11 mm) and large (11-16 mm) sizes. A dependence in relation to temperature was recorded for the ammonium and phosphate excretions that was significantly higher at 25 degrees C than at 20 and 15 degrees C. Through a linear relation between biomass (dry weight) and total length and, between excretion and biomass and, data on chironomids densities, after an intense sampling in 33 sites distributed all along the reservoir bottom, the mean phosphate and ammonium excretion rates corresponded to 2,014 +/- 5,134 microg.m(-2)/day and 1,643 +/- 3,974 microg.m(-2)/day, respectively. Considering the mean biomass (34 mg.m(-2)) of Chironomus, the lake area (88,156 m(2)) and the mean excretion rates, the contribution of benthic chironomids to the internal loads would be 181 KgP and 147 KgN. for the sampling months (October-November 1998). These values showed that the internal loads by excretion from Chironomus larvae correspond to approximately 33% of the external loads of phosphorus in the lake and, in the case of nitrogen, to only 5%.
Subject(s)
Chironomidae/metabolism , Environmental Monitoring , Nitrogen/analysis , Phosphates/metabolism , Phosphorus/analysis , Quaternary Ammonium Compounds/metabolism , Animals , Biomass , Eutrophication , Fresh Water/chemistry , Larva/metabolism , Phosphates/analysis , Population Density , Quaternary Ammonium Compounds/analysis , TemperatureABSTRACT
Measurements of ammonium and phosphate excretion by the Chironomus larvae were conducted in order to evaluate the importance of these chironomids for the internal loads of a small eutrophic urban reservoir. Ammonium and phosphate excretion rates by Chironomus larvae of small size (6-10 mm total length) were significantly higher than those of the Chironomids having medium (9-11 mm) and large (11-16 mm) sizes. A dependence in relation to temperature was recorded for the ammonium and phosphate excretions that was significantly higher at 25 °C than at 20 and 15 °C. Through a linear relation between biomass (dry weight) and total length and, between excretion and biomass and, data on chironomids densities, after an intense sampling in 33 sites distributed all along the reservoir bottom, the mean phosphate and ammonium excretion rates corresponded to 2,014 ± 5,134 µg.m-2/day and 1,643 ± 3,974 µg.m-2/day, respectively. Considering the mean biomass (34 mg.m-2) of Chironomus, the lake area (88,156 m²) and the mean excretion rates, the contribution of benthic chironomids to the internal loads would be 181 KgP and 147 KgN. for the sampling months (October-November 1998). These values showed that the internal loads by excretion from Chironomus larvae correspond to approximately 33 percent of the external loads of phosphorus in the lake and, in the case of nitrogen, to only 5 percent.
Medidas de excreção de amônia e fosfato por larvas de Chironomus foram executadas com a finalidade de avaliar a importância desses quironomídeos nas cargas internas de um pequeno reservatório urbano eutrófico. Taxas de excreção de amônia e fosfato significativamente mais elevadas foram encontradas nas larvas de Chironomus de pequeno (6-10 mm) tamanho, em relação às de tamanho médio (9-11 mm) e grande (11-16 mm). Uma dependência em relação à temperatura foi registrada para a excreção de amônia e fosfato, significativamente mais alta a 25 °C do que a 20 e 15 °C. Depois da obtenção das relações lineares entre biomassa (peso seco) e comprimento total e excreção com biomassa e dos dados de densidade de quironomídeos, após amostragem em 33 pontos distribuídos por todo o reservatório, foi possível estimar uma taxa média de excreção de fosfato de 2014 ± 5134 µg.m-2/dia e, de amônia em 1643 ± 3974 µg.m-2/dia. Considerando a biomassa média (34 mg.m-2) de quironomídeos, a área do lago (88.156 m²) e as taxas médias de excreção, a contribuição dos quironomídeos bênticos para as cargas internas seria de 181 KgP e 147 KgN, para o mês da amostragem (outubro-novembro de 1998). Esses valores mostraram que as cargas internas por excreção pelas larvas de Chironomus correspondem aproximadamente a 33 por cento das cargas externas de fósforo para o lago e, somente 5 por cento no caso de nitrogênio.