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1.
J Helminthol ; 94: e71, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31409433

RESUMO

Human ascariasis is a neglected tropical disease of great relevance to public health and is considered the most frequent helminthiasis in poor regions. Accurately diagnosing this parasite has been challenging due to limitations of current diagnostic methods. Immunoglobulin Y (IgY) technology is a very effective alternative for the production of highly specific and profitable antibodies. This study aimed to produce and apply anti-Ascaris suum IgY antibodies in the immunodiagnosis of human ascariasis. Five immunizations comprising total saline extract from A. suum adult life forms were given at 14-day intervals to Gallus gallus domesticus hens of the Isa Brown line. Eggs and blood samples were collected weekly and fortnightly, respectively, to monitor the production of antibodies. The specificity of antibodies was confirmed by dot-blot, kinetic enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence antibody tests. The application for disease diagnosis was performed through the detection of immune complexes in human serum samples by sandwich ELISA. Peaks of IgY anti-A. suum production occurred at weeks 6 and 8. IgY showed high avidity levels after the second dose of immunization, ranging from 64% to 93%, with a mean avidity index of 78.30%. Purified IgY recognized 12 bands of proteins from A. suum saline extract. Eggs, the uterine portion and cuticles of A. suum female adult are reactive in immunofluorescence. The detection of immune complexes showed diagnostic values of 80% sensitivity and 90% specificity. In conclusion, specific IgY have been shown to be a potential immunodiagnostic tool with promising future applications in human ascariasis.


Assuntos
Anticorpos Anti-Helmínticos/biossíntese , Ascaríase/diagnóstico , Ensaio de Imunoadsorção Enzimática , Imunoglobulinas/biossíntese , Animais , Complexo Antígeno-Anticorpo , Antígenos de Helmintos/administração & dosagem , Antígenos de Helmintos/imunologia , Ascaris suum , Galinhas , Feminino , Humanos , Imunização , Testes Imunológicos/métodos , Sensibilidade e Especificidade
2.
J Med Primatol ; 47(6): 412-415, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-29956824

RESUMO

BACKGROUND: The taxonomic classification of squirrel monkeys is often controversial issue offering many different information. The classification of captive animals is difficult due to the phenotypic similarities between the presented species, which is observed mainly in coat coloration. METHODS: The objective of this study was to analyze the chromosome pattern of one squirrel monkey with off standard physical characteristics, which is kept in the Laboratory Animals Breeding Center in Rio de Janeiro State, Brazil, and try to establish some correlations. Chromosomes were obtained using lymphocyte culture technique. RESULTS AND CONCLUSIONS: Evaluation of G bands showed a terminal deletion in one chromosome of pair 13. The association of the results found with the different phenotypic characteristics led us to classify it as a Saimiri sciureus specimen with a structural chromosomal change, possibly allowing the expression of hemizygous alleles.


Assuntos
Deleção Cromossômica , Cromossomos de Mamíferos/genética , Saimiri/genética , Animais , Animais de Laboratório/genética , Brasil , Masculino , Fenótipo
3.
J Med Primatol ; 46(2): 51-55, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-28185292

RESUMO

Osteosarcoma (OS) is the most common type of bone cancer, especially in young. Telangiectatic osteosarcoma (TO) is a rare variant of OS, and hence, its occurrence, presentation, and prognosis are poorly understood. A 4-year-old female rhesus monkey presenting lameness and swelling was examined for a mass on the right humerus. Radiography revealed fracture and disorganized structure of bone tissue. Histopathological examination revealed malignant neoplasm composed of anaplastic osteoblasts, which invaded the bone marrow and surrounded blood-filled cysts in the epiphysis and diaphysis forming septa. Cytogenetic analysis showed aneuploid cells, supernumerary AgNORs, and a marker fragment. The neoplasm was diagnosed as TO. To our knowledge, the occurrence of TO and its cytogenetic analysis were reported for the first time in non-human primates.


Assuntos
Neoplasias Ósseas/veterinária , Macaca mulatta , Doenças dos Macacos/diagnóstico por imagem , Osteossarcoma/veterinária , Telangiectasia/veterinária , Animais , Neoplasias Ósseas/diagnóstico por imagem , Análise Citogenética/veterinária , Feminino , Osteossarcoma/diagnóstico por imagem , Radiografia/veterinária , Telangiectasia/diagnóstico por imagem
4.
J Appl Microbiol ; 123(2): 450-463, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28543932

RESUMO

AIMS: Evaluating the winemaking stress tolerance of a set of both Saccharomyces eubayanus and Saccharomyces uvarum strains from diverse Patagonian habitats. METHODS AND RESULTS: Yeast strains growth was analysed under increasing ethanol concentrations; all of them were able to grow until 8% v/v ethanol. The effect of different temperature and pH conditions as well as at SO2 and hexose concentrations was evaluated by means of a central composite experimental design. Only two S. uvarum strains (NPCC 1289 and 1321) were able to grow in most stress conditions. Kinetic parameters analysed (µmax and λ) were statistically affected by temperature, pH and SO2 , but not influenced by sugar concentration. The obtained growth model was used for predicting optimal growth conditions for both strains: 20°C, 0% w/v SO2 and pH 4·5. CONCLUSIONS: Strains from human-associated environments (chichas) presented the highest diversity in the response to different stress factors. Two S. uvarum strains from chichas demonstrated to be the most tolerant to winemaking conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This work evidenced the potential use of two S. uvarum yeast strains as starter cultures in wines fermented at low temperatures. Saccharomyces eubayanus was significantly affected by winemaking stress conditions, limiting its use in this industry.


Assuntos
Etanol/metabolismo , Saccharomyces/metabolismo , Vinho/microbiologia , Reatores Biológicos , Fermentação , Saccharomyces cerevisiae/metabolismo , Temperatura , Vinho/análise
5.
J Fish Biol ; 90(4): 1265-1282, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27859226

RESUMO

This study examined the fish communities of Peri Lagoon in southern Brazil to aid in the development of an effective management plan because the area is under threat from human activities. Sampling of fish fauna, ichthyoplankton and limnological data were compared between sites, differing by habitat type and characteristics such as depth, substratum composition and vegetation type. Results were significantly related to site, with the highest diversity and abundance recorded at shallow vegetated sites. A total of 14 fish species were recorded throughout the lagoon, with the most abundant being Hyphessobrycon luetkenii. Of the 14 species, half were sampled at their larval stage, suggesting a healthy and protected system. Significantly more larvae and eggs were collected during colder months (autumn to winter) and at sites closer to stream flow, possibly owing to increased food sources and habitat protection. This study highlights the importance of Peri Lagoon as a nursery ground for a wide range of fish species, providing essential information for incorporation into the future protection of fish stocks throughout Brazil.


Assuntos
Biodiversidade , Peixes/classificação , Água Doce , Água do Mar , Animais , Brasil , Conservação dos Recursos Naturais , Peixes/fisiologia , Óvulo , Estações do Ano
6.
Oral Dis ; 22(4): 303-12, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26788715

RESUMO

OBJECTIVES: To analyse the expression of the CD44v6, p63, podoplanin and MMP-9, and their prognostic significance in patients with oral squamous cell carcinomas (OSCC). MATERIAL AND METHODS: Immunohistochemistry technique was performed on 60 OSCC for detection of CD44v6, p63, podoplanin and MMP-9 proteins. Extent and intensity of staining were evaluated in tumour cells and were compared with patients' clinical-pathological characteristics and survival. RESULTS: CD44v6 expression was detected at the membrane of tumour cells of 94% cases. Nuclear expression of p63 protein was present in 96.5%. Podoplanin was observed at the membrane of tumour cells of 94% cases. MMP-9 was found in the cytoplasm of tumour cells in 83.7% cases. A high level of expression (67%-89%) in all four proteins was noted. Podoplanin was associated with the expression of MMP-9 (P = 0.010) and both were associated with lymph node metastasis (P = 0.011 and P = 0.018, respectively). Co-expression of podoplanin/MMP-9 was an adverse independent prognostic factor for cancer-specific survival (P = 0.008) and recurrence-free survival (P = 0.042). CONCLUSION: Podoplanin and MMP-9 together could contribute to tumour progression and dissemination of OSCC. Their combined overexpression showed an adverse effect on survival, suggesting that they could be regarded as important prognostic biomarkers in OSCC.


Assuntos
Carcinoma de Células Escamosas/química , Receptores de Hialuronatos/análise , Metaloproteinase 9 da Matriz/análise , Glicoproteínas de Membrana/análise , Proteínas de Membrana/análise , Neoplasias Bucais/química , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Taxa de Sobrevida
7.
Oral Dis ; 21(6): 713-20, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25754611

RESUMO

OBJECTIVES: BubR1 and Mad2 are central components of the mitotic checkpoint complex that inhibits anaphase onset until all chromosomes are correctly aligned at the metaphase plate. We propose to analyse the combined expression of BubR1 and Mad2 and assess its significance to oral squamous cell carcinoma (OSCC) diagnosis and prognosis. MATERIALS AND METHODS: BubR1 and Mad2 expression was assessed by real-time PCR in OSCC cell lines and in normal human oral keratinocytes, and by immunohistochemistry in 65 patients with OSCC. The results were compared regarding clinicopathological parameters, proliferative activity and survival. RESULTS: BubR1 and Mad2 transcripts were overexpressed in OSCC cell lines which also exhibited attenuated spindle assembly checkpoint activity. BubR1 and Mad2 were also overexpressed in patients with OSCC. BubR1 expression was associated with advanced stages and larger tumour size in univariate analysis, and with shorter overall survival both in univariate and multivariate analysis. Mad2 overexpression was associated with that of BubR1 and, importantly, high expression of Mad2 and BubR1 was associated with increased cellular proliferation. CONCLUSION: Our data propose a role for BubR1 and Mad2 in OSCC cellular proliferation, progression and prognosis.


Assuntos
Carcinoma de Células Escamosas/química , Proteínas Mad2/análise , Neoplasias Bucais/química , Proteínas Serina-Treonina Quinases/análise , RNA Mensageiro/metabolismo , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Expressão Gênica , Humanos , Queratinócitos/metabolismo , Pontos de Checagem da Fase M do Ciclo Celular/genética , Proteínas Mad2/genética , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/genética , Neoplasias Bucais/patologia , Proteínas Serina-Treonina Quinases/genética , Taxa de Sobrevida
8.
Med Oral Patol Oral Cir Bucal ; 19(2): e120-6, 2014 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-24121907

RESUMO

OBJECTIVES: Our aim was to analyze the clinical, pathological, and outcome characteristics of oral squamous cell carcinomas (OSCC) from a population of the North of Portugal. MATERIAL AND METHODS: We conducted a descriptive study of 128 OSCC diagnosed between the years of 2000 and 2010 in the Centro Hospitalar do Porto. Through of the review of the clinical records we studied several clinical, pathological, and outcome variables. The overall survival (OS) and disease-free survival (DFS) were analyzed by Kaplan-Meier method and log-rank test. Cox regression method was used for multivariate analysis. RESULTS: Of 128 patients with OSCC, 83 (64.8%) were male and 45 (35.2%) were female, (mean age of 62.13±15.57 years). The most affected location was the tongue (n=52; 40.6%). The most common cause of reference was a non-healing ulcer (n=35; 28.9%) followed by oral pain (n=27; 22.3%). Sixty (60.6%) patients were tobacco consumers and 55 (57.3%) alcohol consumers. The cumulative 3-years OS rate was 58.6% and DFS was 55.4%. In multivariable analysis for OS, we found an adverse independent prognostic value for advanced tumour size (p<0.001) and for the presence of perineural permeation (p=0.012). For DFS, advanced stage tumours presented adverse independent prognostic value (p<0.001). CONCLUSION: OSCC occurred most frequently in males, in older patients, and in patients with tobacco and/or alcohol habits. TNM and tumour stage additionally to the perineural permeation were the most important prognostic factor for the survival of these patients, contributing to identify high-risk subgroups and to guide therapy.


Assuntos
Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/patologia , Neoplasias Bucais/mortalidade , Neoplasias Bucais/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Portugal , Prognóstico , Estudos Retrospectivos , Taxa de Sobrevida , Adulto Jovem
9.
J Appl Microbiol ; 114(4): 1066-74, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23311591

RESUMO

AIMS: Dekkera bruxellensis and Pichia guilliermondii are contaminating yeasts in wine due to the production of phenolic aromas. Although the degradation pathway of cinnamic acids, precursors of these phenolic compounds has been described in D. bruxellensis, no such pathway has been described in P. guilliermondii. METHODS AND RESULTS: A molecular and physiological characterization of 14 D. bruxellensis and 15 P. guilliermondii phenol-producing strains was carried out. Both p-coumarate decarboxylase (CD) and vinyl reductase (VR) activities, responsible for the production of volatile phenols, were quantified and the production of 4-vinylphenol and 4-ethylphenol were measured. All D. bruxellensis and some P. guilliermondii strains showed the two enzymatic activities, whilst 11 of the 15 strains of this latter species showed only CD activity and did not produce 4-EP in the assay conditions. Furthermore, PCR products obtained with degenerated primers showed a low homology with the sequence of the gene for a phenyl acrylic acid decarboxylase activity described in Saccharomyces cerevisiae. CONCLUSIONS: D. bruxellensis and P. guilliermondii may share a similar metabolic pathway for the degradation of cinnamic acids. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first work that analyses the CD and VR activities in P. guilliermondii, and the results suggest that within this species, there are differences in the metabolization of cinnamic acids.


Assuntos
Cinamatos/metabolismo , Dekkera/metabolismo , Pichia/metabolismo , Vinho/microbiologia , Carboxiliases/metabolismo , Dekkera/genética , Contaminação de Alimentos , Oxirredutases/metabolismo , Fenóis/análise , Pichia/genética , Reação em Cadeia da Polimerase , Técnica de Amplificação ao Acaso de DNA Polimórfico
10.
Reprod Fertil Dev ; 25(6): 955-65, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23050662

RESUMO

The aim of the present study was to evaluate the effects of growth differentiation factor 9 (GDF-9) and FSH on the in vitro development of caprine preantral follicles cultured for 16 days. Ovarian fragments were cultured in αMEM⁺ (α-minimum essential medium, pH 7.2-7.4, 10 µg mL⁻¹ insulin, 5.5 µg mL⁻¹ transferrin, 5.0 ng mL⁻¹ selenium, 2 mM glutamine, 2 mM hypoxanthine and 1.25 mg mL⁻¹ bovine serum albumin) in the absence or presence of 200 ng mL⁻¹ GDF-9 and/or 50 ng mL⁻¹ FSH added during the first (Days 0-8) and/or second (Days 8-16) half of the culture period. Non-cultured and cultured fragments were processed for histological and ultrastructural analyses. After 16 days, all treatments using GDF-9 or FSH showed higher rates of follicular survival compared with αMEM⁺ alone. Compared with non-cultured control, sequential culture media containing GDF-9 and/or FSH significantly increased the percentage of developing follicles and follicle diameter. Moreover, a progressive increase in oocyte diameter was observed only with sequential culture medium containing GDF-9 until Day 8 followed by FSH (GDF-9/FSH) in the second half of the culture period. After 16 days of culture, ultrastructural analysis confirmed the integrity of follicles cultured in the presence of GDF-9/FSH. In conclusion, a dynamic medium containing GDF-9 and FSH (GDF-9/FSH) maintained follicular integrity and promoted activation of primordial follicles and growth during long-term in vitro culture of goat preantral follicles.


Assuntos
Fármacos para a Fertilidade Feminina/farmacologia , Hormônio Foliculoestimulante/farmacologia , Cabras/fisiologia , Fator 9 de Diferenciação de Crescimento/farmacologia , Técnicas de Maturação in Vitro de Oócitos/veterinária , Folículo Ovariano/efeitos dos fármacos , Matadouros , Animais , Brasil , Bovinos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cruzamentos Genéticos , Feminino , Hormônio Foliculoestimulante/genética , Fator 9 de Diferenciação de Crescimento/genética , Humanos , Oócitos/efeitos dos fármacos , Oócitos/fisiologia , Oócitos/ultraestrutura , Oogênese/efeitos dos fármacos , Folículo Ovariano/fisiologia , Folículo Ovariano/ultraestrutura , Proteínas Recombinantes/farmacologia , Fatores de Tempo , Técnicas de Cultura de Tecidos/veterinária
11.
Cells Tissues Organs ; 195(3): 260-71, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-21494023

RESUMO

The aim of this study was to evaluate the effects of a dynamic medium containing kit ligand (KL) and follicle-stimulating hormone (FSH) on the in vitro culture of caprine preantral follicles for 16 days. Ovarian fragments were cultured in α-MEM(+) containing or not containing KL (50 ng/ml) and/or FSH (50 ng/ml) added during the first (days 0-8) and/or second half (days 8-16) of the culture period. Noncultured (control) and cultured fragments were processed for histological and ultrastructural evaluation. After 1 day of culture, only the treatments performed with KL or FSH maintained a percentage of normal follicles similar to that of the control. After 16 days, all treatments using KL until day 8 (KL/KL, KL/FSH, and KL/FSH+KL) and only FSH during the entire culture period (FSH/FSH) showed higher rates of follicular survival compared to α-MEM(+) alone. After 1 and 8 days, the treatments initially cultured with KL increased the percentage of follicular activation in comparison to α-MEM(+) alone and other treatments. The highest follicular diameter after 16 days was observed in follicles cultured with KL until day 8 followed by FSH (KL/FSH). Furthermore, this treatment promoted, as early as after 1 day of culture, an increase in oocyte growth compared to α-MEM(+) alone. Ultrastructural analysis confirmed the integrity of follicles cultured in KL/FSH after 16 days. In conclusion, a dynamic medium containing KL and FSH maintained follicular integrity and promoted follicular activation and growth during the long-term in vitro culture of caprine preantral follicles.


Assuntos
Hormônio Foliculoestimulante/farmacologia , Cabras/fisiologia , Folículo Ovariano/efeitos dos fármacos , Fator de Células-Tronco/farmacologia , Animais , Meios de Cultura , Feminino , Humanos , Microscopia de Fluorescência , Oócitos/citologia , Oócitos/efeitos dos fármacos , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Folículo Ovariano/citologia , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/metabolismo
12.
Cell Tissue Res ; 346(3): 451-6, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22089148

RESUMO

This study investigated the effect of adding different insulin concentrations to the culture medium for goat preantral follicle development in vitro. The ovarian fragments were immediately fixed or cultured for 7 days in MEM with insulin (0, 5, 10 ng/ml and 5 or 10 µg/ml). The results showed that, after 7 days of culture, insulin at 10 ng/ml was the best concentration to preserve follicular viability and ultrastructure, resulting in the highest rates of normal follicles. After 7 days, only treatments with 10 ng/ml and 5 µg/ml of insulin increased follicular activation when compared to other concentrations. Regarding follicular and oocyte growth, the presence of 10 ng/ml of insulin promoted a larger diameter than other treatments. In conclusion, this study shows that addition of 10 ng/ml of insulin to the culture medium improved the survival and stimulated growth of goat preantral follicles.


Assuntos
Insulina/farmacologia , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/crescimento & desenvolvimento , Animais , Feminino , Cabras
13.
Cells Tissues Organs ; 192(4): 272-82, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20587997

RESUMO

The aim of this study was to investigate the effects of nerve growth factor (NGF) on the in vitro culture of goat preantral follicles. Ovarian cortex fragments were cultured in α-MEM+ supplemented with 0, 1, 10, 50, 100 or 200 ng/ml NGF for 1 or 7 days. Small fragments of noncultured ovarian tissue as well as those cultured for 1 or 7 days were processed for histology and transmission electron microscopy. The results showed that after 1 or 7 days of culture at all concentrations of NGF, except at 1 ng/ml after 1 day of culture, there was a significant reduction in the percentage of normal follicles compared to noncultured tissues. At higher NGF concentrations (100 and 200 ng/ml) after 7 days of culture, there was a significant reduction in the percentage of normal follicles compared to tissues cultured in α-MEM+ alone or at the other concentrations of NGF. It is important to note that ultrastructural and fluorescent analyses confirmed only the integrity of follicles cultured with 1 ng/ml of NGF after 7 days. In contrast to noncultured control tissues, the percentage of developing follicles was significantly increased at all concentrations of NGF after 1 or 7 days of culture. We observed that follicular diameter was greater at 1 and 10 ng/ml NGF after culture for 7 days than at the other concentrations but was similar to follicles cultured in α-MEM+ alone. In conclusion, NGF improved the survival of goat preantral follicles cultured in vitro in a dose-dependent manner.


Assuntos
Fator de Crescimento Neural/farmacologia , Folículo Ovariano , Ovário , Animais , Feminino , Cabras , Microscopia de Fluorescência , Oócitos/efeitos dos fármacos , Oócitos/fisiologia , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/embriologia , Folículo Ovariano/fisiologia , Ovário/efeitos dos fármacos , Ovário/embriologia , Ovário/metabolismo , Receptores de Fator de Crescimento Neural/metabolismo , Técnicas de Cultura de Tecidos
14.
J Appl Microbiol ; 109(6): 2206-13, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20860771

RESUMO

AIMS: The purpose of this study was to characterize the α-l-rhamnosidase of Pichia guilliermondii NPCC1053 indigenous wine strain from North-Patagonian region. METHODS AND RESULTS: The optimization of yeast culture conditions was carried out and the effects of oenological parameters on α-l-rhamnosidase activity were evaluated. Additionally, the effect of direct contact with must and wine on α-l-rhamnosidase activity was assayed. This strain showed an intracellular inducible α-l-rhamnosidase activity. This enzyme was active at pH, glucose and SO(2) concentrations usually found at the beginning of the fermentation as well as retained high levels of activity after 24 h of incubation in must. Furthermore, P. guilliermondiiα-l-rhamnosidase was able to release monoterpenols and alcohols from grape glycosidic extracts. CONCLUSIONS: The α-l-rhamnosidase belonging to P. guilliermondii indigenous wine yeast strain showed mainly an intracellular location and evidenced interesting oenological characteristics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to the knowledge of α-l-rhamnosidases from yeast origin because at present, there are few reports about this enzymatic activity in these micro-organisms. In addition, this work is relevant to the regional wine industry considering that this enzyme could be used in the production of more aromatic young wines.


Assuntos
Glicosídeo Hidrolases/metabolismo , Pichia/enzimologia , Vinho/microbiologia , Argentina , Meios de Cultura , Fermentação , Pichia/crescimento & desenvolvimento
15.
Lett Appl Microbiol ; 51(2): 170-6, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20565575

RESUMO

AIMS: To investigate whether the presence of Pichia guilliermondii impacts on the production of volatile phenols from mixed wine fermentations with Dekkera bruxellensis and Saccharomyces cerevisiae. METHODS AND RESULTS: Four inoculation strategies were performed in small-scale fermentations involving P. guilliermondii, D. bruxellensis and S. cerevisiae using Syrah grape juice supplemented with 100 mg l(-1) of p-coumaric acid. High pressure liquid chromatography was used for the quantification or volatile phenols. Significant high levels of 4-ethylphenol and 4-ethylguaicol (720 and 545 microg l(-1), respectively), as well as the highest levels of 4-vinylphenol (>4500 microg l(-1)), were observed when P. guilliermondii species was inoculated from the beginning of the fermentation. CONCLUSIONS: The metabolic interaction occurring between the high vinylphenol producer species P. guilliermondii and D. bruxellensis exhibiting a high vinylphenol reductase activity resulted in an increased production of volatile phenols in wine. SIGNIFICANCE AND IMPACT OF THE STUDY: Pichia guilliermondii must be considered a very important spoilage yeast in the wine industry capable of producing large amounts of volatile phenols.


Assuntos
Dekkera/metabolismo , Fenóis/análise , Pichia/metabolismo , Saccharomyces cerevisiae/metabolismo , Compostos Orgânicos Voláteis/análise , Vinho/microbiologia , Cromatografia Líquida de Alta Pressão , Dekkera/crescimento & desenvolvimento , Pichia/crescimento & desenvolvimento , Saccharomyces cerevisiae/crescimento & desenvolvimento
16.
Rev Argent Microbiol ; 42(4): 298-306, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21229201

RESUMO

A new optimized semiquantitative yeast killer assay is reported for the first time. The killer activity of 36 yeast isolates belonging to three species, namely, Metschnikowia pulcherrima, Wickerhamomyces anomala and Torulaspora delbrueckii, was tested with a view to potentially using these yeasts as biocontrol agents against the wine spoilage species Pichia guilliermondii and Pichia membranifaciens. The effectiveness of the classical streak-based (qualitative method) and the new semiquantitative techniques was compared. The percentage of yeasts showing killer activity was found to be higher by the semiquantitative technique (60%) than by the qualitative method (45%). In all cases, the addition of 1% NaCl into the medium allowed a better observation of the killer phenomenon. Important differences were observed in the killer capacity of different isolates belonging to a same killer species. The broadest spectrum of action was detected in isolates of W. anomala NPCC 1023 and 1025, and M. pulcherrima NPCC 1009 and 1013. We also brought experimental evidence supporting the importance of the adequate selection of the sensitive isolate to be used in killer evaluation. The new semiquantitative method proposed in this work enables to visualize the relationship between the number of yeasts tested and the growth of the inhibition halo (specific productivity). Hence, this experimental approach could become an interesting tool to be taken into account for killer yeast selection protocols.


Assuntos
Meios de Cultura/farmacologia , Microbiologia Industrial/métodos , Fatores Matadores de Levedura/análise , Testes de Sensibilidade Microbiana/métodos , Micologia/métodos , Vinho/microbiologia , Leveduras/isolamento & purificação , Relação Dose-Resposta a Droga , Fermentação , Controle Biológico de Vetores , Tolerância ao Sal , Cloreto de Sódio/farmacologia , Leveduras/efeitos dos fármacos , Leveduras/metabolismo
17.
Rev Argent Microbiol ; 41(3): 177-84, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19831317

RESUMO

Yeasts belonging to the genus Dekkera/Brettanomyces, especially the species Dekkera bruxellensis, have long been associated with the production of volatile phenols responsible for off-flavour in wines. According to recent reports, the species Pichia guilliermondii could also produce these compounds at the initial stages of fermentation. Based on the abundance of P. guilliermondii in Patagonian winemaking, we decided to study the relevance of indigenous isolates belonging to this species as wine spoilage yeast. Twenty-three indigenous isolates obtained from grape surfaces and red wine musts were analyzed in their capacity to produce volatile phenols on grape must. The relationship between molecular Random Amplified Polymorphic DNA (RAPD) and physiological (killer biotype) patterns detected in indigenous populations of P. guilliermondii and volatile phenol production was also evaluated. Different production levels of 4-ethylphenol, 4-vinylguaiacol and 4-ethylguaiacol were detected among the isolates; however, the values were always lower than those produced by the D. bruxellensis reference strain in the same conditions. High levels of 4-vinylphenol were detected among P. guilliermondii indigenous isolates. The combined use of RAPD and killer biotype allowed us to identify the isolates producing the highest volatile phenol levels.


Assuntos
Fenóis/análise , Pichia/isolamento & purificação , Compostos Orgânicos Voláteis/análise , Vinho/microbiologia , Argentina , Dekkera/metabolismo , Fermentação , Guaiacol/análogos & derivados , Guaiacol/análise , Fatores Matadores de Levedura/farmacologia , Técnicas de Tipagem Micológica , Pichia/efeitos dos fármacos , Pichia/genética , Pichia/metabolismo , Reação em Cadeia da Polimerase , Técnica de Amplificação ao Acaso de DNA Polimórfico , Vitis/microbiologia , Vinho/análise
18.
Ann Biomed Eng ; 47(4): 1153-1164, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30710185

RESUMO

This study analyzes the effects of the Rapha® system, which employs red light therapy (RLT) and a natural latex biomembrane in the healing of neuropathic ulcers associated with the diabetic foot. We conducted a randomized, controlled, blinded clinical trial with 15 participants that were divided into three groups (GI, GII and GIII): (i) Rapha® system application by the participant and a health professional at home, with clinical status evaluation every 2 weeks at the hospital (GI); (ii) standard protocol used in Brazil, performed by a health professional at the hospital (GII; control); and (iii) the Rapha® system applied by the participant at home and clinical status evaluation every 2 weeks at the hospital (GIII). We used image processing techniques on photographic recordings of the lesions, and several statistical tests were used to analyze the data, allowing for the comparison of the average results for all groups. The average healing rates of GI, GII, and GIII were 77.0, 51.4, and 80%, respectively. The granulation tissue evaluation indicated a higher efficacy in the tissue repair of lesions treated with the Rapha® system. In conclusion, the Rapha® system proved to be an effective healing system, even when self-applied at the patient's home.


Assuntos
Bandagens , Pé Diabético/terapia , Látex , Membranas Artificiais , Fototerapia , Cicatrização , Idoso , Pé Diabético/patologia , Pé Diabético/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
19.
Reprod Fertil Dev ; 20(5): 640-7, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18577361

RESUMO

The aim of the present study was to evaluate the effects of storage of goat ovarian fragments at different temperatures and for different incubation times on the viability and growth of cultured preantral follicles in vitro. Caprine ovaries were collected and divided into 19 fragments, with one fragment being fixed immediately (fresh control). The remaining fragments were placed in minimal essential medium (MEM) and maintained at 4, 20 or 35 degrees C for 2 or 4 h. After each incubation period, some of the fragments were fixed (non-cultured), whereas others were cultured in vitro for 1 or 7 days. Fragments were processed to enable routine histological and transmission electron microscopic examination. After 7 days of culture, only ovarian fragments stored at 4 degrees C for 4 h maintained a percentage of morphologically normal follicles similar to that in the fresh control. For all other treatments groups, there was a significant increase in follicular activation observed. In addition, there was an increase in oocyte and follicular diameter after culture of ovarian cortex that had been chilled previously at 4 degrees C for 2 or 4 h. In conclusion, the present study demonstrated that chilling ovarian fragments at 4 degrees C during transportation is best for maintaining follicle viability and to increase follicular growth during in vitro culture.


Assuntos
Temperatura Baixa , Cabras/fisiologia , Recuperação de Oócitos/métodos , Oócitos/crescimento & desenvolvimento , Folículo Ovariano/fisiologia , Meios de Transporte , Animais , Tamanho Celular , Sobrevivência Celular , Células Cultivadas , Feminino , Oócitos/citologia , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/ultraestrutura , Ovário/fisiologia , Temperatura
20.
Braz J Biol ; 78(3): 477-486, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29185610

RESUMO

We evaluated the reproductive dynamics of two fish species, Lycengraulis grossidens and Platanichthys platana, in a subtropical freshwater coastal lagoon (Peri Lagoon) in Brazil. Samples were collected from nine sites every two months from June 2008 to April 2012. Different fishing methods were used to capture larvae, juveniles, and adults. Limnological variables were obtained using multiparameter probe. More females than males were collected of both fish species and the chi-square test (χ2) was used to confirm that the sex ratio was female-biased. Large numbers of maturing and mature fishes were observed in almost every sampling month. The gonadosomatic index (GSI) of L. grossidens was the highest in August, October, and December for females and in October for males, and no significant difference was found across years. The GSI of P. platana showed no significant difference across months for either sex; however, significant differences were recorded in year three (June 2010-April 2011) and year four (June 2011-April 2012) for females and in year three (June 2010-April 2011) for males. For both species, GSI was negatively correlated with temperature and water level. L. grossidens larvae were more abundant in October 2010, showing a positive correlation with water transparency, whereas P. platana larvae were more abundant in June 2011, showing a positive correlation with water transparency and negative correlation with temperature and precipitation. Both species were represented by different stages, including larvae, which confirm that these species reproduce in Peri Lagoon. Reproduction was more pronounced in autumn and winter; however, reproductive activity was evident throughout the sampling period. In conclusion, our results show that abiotic factors strongly influence the temporal pattern of reproductive activity and larval assemblages of both L. grossidens and P. platana in Peri Lagoon, Brazil.


Assuntos
Água Doce , Reprodução , Estações do Ano , Animais , Brasil , Feminino , Peixes , Masculino , Razão de Masculinidade , Temperatura
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