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1.
Tech Coloproctol ; 24(3): 215-220, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31965400

RESUMO

BACKGROUND: The aim of this study was to report the outcomes of sacral neuromodulation (SNM) in chronic pelvic pain (CPP) patients in the Finnish national cohort. METHODS: This was a register-based retrospective study, involving all the centers that provide SNM treatment in Finland. The data of all patients treated with SNM for CPP were gathered from Oulu-, Turku-, Tampere- and Helsinki University Hospitals, as well as Jyväskylä and Seinäjoki Central Hospitals. All patients who had been tested for SNM implantation prior to April 2017 were included in the study. RESULTS: A total of 51 patients were selected for SNM treatment due to CPP from 2004 until 2017. The mean follow-up time was 13.8 months (SD 22.9 months). A total of 28 patients (57%) advanced from testing to permanent stimulator implantation. There were 21 patients (41%) who had a working modulator implanted at the end of follow-up. Patients with endometriosis-related pain had a significantly higher permanent implantation rate than the overall implantation rate (88% vs. 57%; p = 0.01). The endometriosis patients also had a higher overall success rate by the end of the follow-up (75% vs. 41%; p = 0.026) CONCLUSIONS: SNM may be a viable treatment option for patients with CPP due to endometriosis. Further research on SNM treatment for endometriosis patients with refractory CPP is needed.


Assuntos
Terapia por Estimulação Elétrica , Plexo Lombossacral , Eletrodos Implantados , Feminino , Finlândia , Humanos , Dor Pélvica/etiologia , Dor Pélvica/terapia , Estudos Retrospectivos , Sacro , Resultado do Tratamento
2.
Bioelectrochemistry ; 120: 27-42, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29154214

RESUMO

In this work, six common stainless steel grades were compared with respect to ennoblement characteristics, corrosion performance and tendency to biofouling in brackish sea water in a pilot-scale cooling water circuit. Two tests were performed, each employing three test materials, until differences between the materials were detected. Open circuit potential (OCP) was measured continuously in situ. Potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) measurements were conducted before and after the tests. Exposed specimens were further subjected to examinations by scanning electron microscopy (SEM) and energy-dispersive spectroscopy (EDS), and the biofouling was studied using epifluorescence microscopy, quantitative polymerase chain reaction (qPCR) and high-throughput sequencing (HTP sequencing). The results revealed dissimilarities between the stainless steel grades in corrosion behaviour and biofouling tendency. The test material that differed from the most of the other studied alloys was grade EN 1.4162. It experienced fastest and most efficient ennoblement of OCP, its passive area shrank to the greatest extent and the cathodic reaction was accelerated to a significant degree by the development of biofilm. Furthermore, microbiological analyses revealed that bacterial community on EN 1.4162 was dominated by Actinobacteria, whereas on the other five test materials Proteobacteria was the main bacterial phylum.


Assuntos
Incrustação Biológica , Aço Inoxidável/química , Actinobacteria/isolamento & purificação , Corrosão , Espectroscopia Dielétrica , Proteobactérias/isolamento & purificação , Água do Mar/química , Água do Mar/microbiologia
3.
J Environ Radioact ; 178-179: 343-348, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28947086

RESUMO

Uranium is naturally present in seawater at trace levels and may in some cases be present at higher concentrations, due to anthropogenic nuclear activities. Understanding uranium speciation in seawater is thus essential for predicting and controlling its behavior in this specific environmental compartment and consequently, its possible impact on living organisms. The carbonato calcic complex Ca2UO2(CO3)3 was previously identified as the main uranium species in natural seawater, together with CaUO2(CO3)32-. In this work, we further investigate the role of the alkaline earth cation in the structure of the ternary uranyl-carbonate complexes. For this purpose, artificial seawater, free of Mg2+ and Ca2+, using Sr2+ as a spectroscopic probe was prepared. Combining TRLIF and EXAFS spectroscopy, together with DFT and theoretical thermodynamic calculations, evidence for the presence of Sr alkaline earth counter ion in the complex structure can be asserted. Furthermore, data suggest that when Ca2+ is replaced by Sr2+, SrUO2(CO3)32- is the main complex in solution and it occurs with the presence of at least one monodentate carbonate in the uranyl coordination sphere.


Assuntos
Carbonatos/análise , Modelos Químicos , Água do Mar/química , Urânio/análise , Adsorção
4.
Bioresour Technol ; 169: 88-95, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25033328

RESUMO

Fibres fractionated from solid recovered fuel (SRF), a standardised market combustion fuel produced from sorted waste, were considered as a source of lignocellulosic fermentable sugars. The fibre yield from four samples of SRF was 25-45%, and the separated material consisted of 52-54% carbohydrates, mainly glucan, with a high content of ash (12-17%). The enzymatic digestibility of recovered fibres was studied at low and high solids loading and compared with model substrates containing only chemical and mechanical pulps. Above 80% hydrolysis yield was reached at 20% solids loading in 48 h, but variation was observed between different samples of recovered fibres. Surfactants were found to improve the hydrolysis yield of recovered fibres especially in tumbling-type of mixing at low solids loading, where hydrolysis was found to stagnate without surfactants. The results suggest that SRF is a potential source of easily digestible lignocellulosic carbohydrates for use in biorefineries.


Assuntos
Biocombustíveis , Fracionamento Químico/métodos , Hidrólise , Papel , Estresse Mecânico , Tensoativos/química , Temperatura , beta-Glucosidase/metabolismo
5.
Transl Psychiatry ; 1: e57, 2011 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-22832349

RESUMO

Mild cognitive impairment (MCI) is considered as a transition phase between normal aging and Alzheimer's disease (AD). MCI confers an increased risk of developing AD, although the state is heterogeneous with several possible outcomes, including even improvement back to normal cognition. We sought to determine the serum metabolomic profiles associated with progression to and diagnosis of AD in a prospective study. At the baseline assessment, the subjects enrolled in the study were classified into three diagnostic groups: healthy controls (n=46), MCI (n=143) and AD (n=47). Among the MCI subjects, 52 progressed to AD in the follow-up. Comprehensive metabolomics approach was applied to analyze baseline serum samples and to associate the metabolite profiles with the diagnosis at baseline and in the follow-up. At baseline, AD patients were characterized by diminished ether phospholipids, phosphatidylcholines, sphingomyelins and sterols. A molecular signature comprising three metabolites was identified, which was predictive of progression to AD in the follow-up. The major contributor to the predictive model was 2,4-dihydroxybutanoic acid, which was upregulated in AD progressors (P=0.0048), indicating potential involvement of hypoxia in the early AD pathogenesis. This was supported by the pathway analysis of metabolomics data, which identified upregulation of pentose phosphate pathway in patients who later progressed to AD. Together, our findings primarily implicate hypoxia, oxidative stress, as well as membrane lipid remodeling in progression to AD. Establishment of pathogenic relevance of predictive biomarkers such as ours may not only facilitate early diagnosis, but may also help identify new therapeutic avenues.


Assuntos
Doença de Alzheimer/metabolismo , Disfunção Cognitiva/metabolismo , Progressão da Doença , Via de Pentose Fosfato/fisiologia , Idoso , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/patologia , Biomarcadores/sangue , Biomarcadores/metabolismo , Disfunção Cognitiva/diagnóstico , Disfunção Cognitiva/patologia , Feminino , Seguimentos , Humanos , Masculino , Metaboloma/fisiologia , Valor Preditivo dos Testes
6.
Euro Surveill ; 15(45)2010 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-21087590

RESUMO

In September 2009, an outbreak of 2009 pandemic influenza A(H1N1) took place in a Finnish garrison. In November 2009, we performed a serological survey among 984 recruits undergoing their military service at the garrison and related the results to self-reported upper respiratory tract infection (URTI) with or without fever. Of 346 volunteers who donated a blood sample, 169 (49%) had pandemic influenza A(H1N1) virus-specific antibodies. Of those, 84 (50%) reported no recent history of URTI, suggesting that a major part of those infected with pandemic influenza A(H1N1) virus may be asymptomatic.


Assuntos
Anticorpos Antivirais/sangue , Vírus da Influenza A Subtipo H1N1 , Influenza Humana/epidemiologia , Militares , Pandemias/estatística & dados numéricos , Adulto , Reações Cruzadas , Feminino , Febre/etiologia , Finlândia/epidemiologia , Testes de Inibição da Hemaglutinação , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/imunologia , Influenza Humana/imunologia , Influenza Humana/virologia , Masculino , Prevalência , Infecções Respiratórias/epidemiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Autorrelato , Estudos Soroepidemiológicos , Inquéritos e Questionários , Adulto Jovem
7.
Med Trop (Mars) ; 70(1): 43-8, 2010 Feb.
Artigo em Francês | MEDLINE | ID: mdl-20337114

RESUMO

PURPOSE: To evaluate the efficacy of Widal-Felix serodiagnostic testing in Togo. METHOD: This study using a cohort of 200 patients recruited at hospitals in Lome and Atakpame from November 2005 to April 2006 was designed to compare the sensitivity and specificity of the routinely used technique (plate agglutination) and reference technique (tube agglutination). RESULTS: Findings showed that the prevalence of typhoid fever was 1.5% while that of paratyphoid fever was nil. The hardest-hit age group was young people between 11 to 20 years. Men were two times more likely than women to be infected (sex-ratio, 0.5). The most useful symptoms for discriminating patients seropositive for Salmonella typhi from other serological profiles were abdominal pain (p<0.034) and diarrhea (p<0.008). The thick-drop malaria test was positive in all 3 patients (100%) with proven typhoid fever in comparison with 3.9% of the 154 patients with negative Widal-Felix serodiagnostic tests and 26.2% of the 42 patients with intermediate serological profiles. The sensitivity and specificity of the plate technique were 60.0% and 98.06% respectively. The sensitivity and specificity of laboratory analysis were 60.0% and 91.08% respectively. CONCLUSION: The gap between the specificity of the routine plate technique and laboratory analysis underscores the difficulty of performing and interpreting the Widal-Felix serodiagnostic test. The low sensitivity and specificity of the Widal-Felix serodiagnostic test also cast doubt on systematic use for diagnosis in patients presenting fever and on initiation of antibiotic treatment based on agglutination of a single antigen. To improve the specificity of the Widal-Felix serodiagnostic test, we recommend standardization of interpretation criteria and use of tube agglutination. We also see the need for development of another reproducible immunologic test for the diagnosis of typhoid and paratyphoid infections.


Assuntos
Testes de Aglutinação , Febre Tifoide/diagnóstico , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Sensibilidade e Especificidade , Togo , Adulto Jovem
8.
Nutr Metab Cardiovasc Dis ; 20(4): 249-57, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-19553094

RESUMO

BACKGROUND AND AIMS: Whole-grain cereals and diets with a low glycemic index may protect against the development of type 2 diabetes and heart disease, but the mechanisms are poorly understood. We studied the effect of carbohydrate modification on serum metabolic profiles, including lipids and branched chain amino acids, and dependencies between these and specific gene expression pathways in adipose tissue. METHODS AND RESULTS: Twenty subjects with metabolic syndrome were selected from the larger FUNGENUT study population, randomized either to a diet high in oat and wheat bread and potato (OWP) or rye bread and pasta (RP). Serum metabolomics analyses were performed using ultra-performance liquid chromatography coupled to electrospray ionization mass spectrometry (UPLC/MS), gas chromatography (GC) and UPLC. In the OWP group multiple proinflammatory lysophosphatidylcholines increased, while in the RP group docosahexaenoic acid (DHA 22:6n-3) increased and isoleucine decreased. mRNA expression of stress reactions- and adipose tissue differentiation-related genes were up-regulated in adipose tissue in the OWP group. In the RP group, however, pathways related to stress reactions and insulin signaling and energy metabolism were down-regulated. The lipid profiles had the strongest association with the changes in the adipose tissue differentiation pathway when using the elastic net regression model of the lipidomic profiles on selected pathways. CONCLUSION: Our results suggest that the dietary carbohydrate modification alters the serum metabolic profile, especially in lysoPC species, and may, thus, contribute to proinflammatory processes which in turn promote adverse changes in insulin and glucose metabolism.


Assuntos
Carboidratos da Dieta/farmacologia , Síndrome Metabólica/sangue , Síndrome Metabólica/dietoterapia , Tecido Adiposo/metabolismo , Tecido Adiposo/fisiologia , Aminoácidos de Cadeia Ramificada/sangue , Vias Biossintéticas , Cromatografia Líquida de Alta Pressão , Dieta , Ingestão de Alimentos , Ácidos Graxos/sangue , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/genética , Índice Glicêmico , Humanos , Lipídeos/sangue , Síndrome Metabólica/genética , Metabolômica , Dobras Cutâneas , Espectrometria de Massas por Ionização por Electrospray , Resultado do Tratamento
9.
Food Chem ; 110(1): 168-76, 2008 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-26050180

RESUMO

The water-insolubility of some coffee extract components is one of the major limitations in the production of instant coffee. In this work, fractions from coffee extracts and sediments were prepared, and their chemical composition determined. Based on the carbohydrate analysis, galactomannan was found to be the main polysaccharide component of the insoluble fractions and probably responsible for sediment formation. The suitability of twelve commercial enzymes for the hydrolysis of the insoluble fractions was investigated. Pectinase 444L was the most effective enzyme in releasing sugars, mainly mannose and galactose, from these substrates. Biopectinase CCM, Rohapect B1L, Pectinase 444L and Galactomannanase ACH were found to be the most effective enzymes for reducing the sediment of coffee extracts. The highest sediment reduction was obtained using Rohapect B1L and Galactomannanase ACH, at enzyme concentrations of 0.3 and 0.1mg protein/g substrate, respectively.

10.
J Ind Microbiol Biotechnol ; 33(5): 359-67, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16418870

RESUMO

In this study, polysaccharide-producing bacteria were isolated from slimes collected from two Finnish and one Spanish paper mill and the exopolysaccharides (EPSs) produced by 18 isolates were characterised. Most of the isolates, selected on the bases of slimy colony morphology, were members of the family Enterobacteriaceae most frequently belonging to the genera Enterobacter and Klebsiella including Raoultella. All of the EPSs analysed showed the presence of charged groups in the form of uronic acid or pyruvate revealing the polyanionic nature of these polysaccharides. Further results of the carbohydrate analysis showed that the EPS produced by nine of the enterobacteria was colanic acid.


Assuntos
Biofilmes , Enterobacteriaceae/metabolismo , Papel , Polissacarídeos/metabolismo , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Indústrias , Filogenia
11.
J Ind Microbiol Biotechnol ; 32(3): 109-14, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15750806

RESUMO

Development of novel enzymatic methods for slime deposit control in paper mills requires knowledge of polysaccharide-producing organisms and the polysaccharide structures present in deposits. In this work, 27 polysaccharide-producing bacteria were isolated from slime samples collected from different parts of a paper machine. Most of the isolates produced polysaccharides in liquid culture and nine of them were selected for production of polysaccharides for characterisation. The selected isolates belonged to seven different genera: Bacillus, Brevundimonas, Cytophaga, Enterobacter, Klebsiella, Paenibacillus and Starkeya. Using ribotyping, partial 16S rDNA sequencing, physiological tests and fatty acid analysis, four of the nine isolates: Bacillus cereus, Brevundimonas vesicularis, K. pneumoniae and P. stellifer were identified to the species level. Production of polysaccharides by the selected isolates varied between 0.07 and 1.20 g L(-1), the highest amount being produced by B. vesicularis. The polysaccharides were heteropolysaccharides with varying proportions of galactose, glucose mannose, rhamnose fucose and uronic acids.


Assuntos
Bactérias/metabolismo , Biofilmes , Microbiologia Industrial , Papel , Polissacarídeos Bacterianos/metabolismo , Bactérias/genética , DNA Bacteriano/análise
12.
Acta Anaesthesiol Scand ; 49(1): 62-5, 2005 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15675984

RESUMO

BACKGROUND: The number of intensive care units (ICU) using a clinical information system (CIS) is increasing. It is believed that replacing manual charting with an automatic documentation system allocates nurses more time for patient care. The objective of this study was to measure changes in nurses' working time utilization after the implementation of a CIS in a polyvalent ICU of a large Finnish central hospital. METHODS: An activity analysis-based comparison of the ICU nurses' working time utilization before and after the implementation of a CIS. RESULTS: After the implementation of a CIS the total time the nurses spent on documentation of nursing care increased by 3.6% (NS), 15 min per shift of 8 h per nurse. The total time they spent on patient care increased by 5.5% (P < 0.05), 21 min. Intensive care nursing activities increased by 3.7% (P < 0.05), 14 min. The length of the nurses' ICU experience had some effect on these figures. The demand for nurse labor remained constant. CONCLUSIONS: After the implementation of a CIS, an increase in the time nurses spent on documentation of care was detected, which suggests a need for further development of the system. As all the measured time changes were relatively small, any plans to reduce the ICU staff number with the aid of computers were not justified.


Assuntos
Sistemas de Informação em Laboratório Clínico , Documentação , Unidades de Terapia Intensiva/organização & administração , Enfermeiras e Enfermeiros , Cuidados Críticos , Seguimentos , Estudos de Tempo e Movimento , Recursos Humanos
13.
Lett Appl Microbiol ; 39(4): 336-40, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15355535

RESUMO

AIMS: The aim was to develop a cheap cereal-based alternative medium for the large-scale production of biopreservative Lactobacillus plantarum VTT E-79098. We examined the effect of growth medium and pH control on the cell yield of Lact. plantarum E-79098 and the antimicrobial activity of the cell-free extracts. METHODS: Fermentations using a novel Malt Sprout Extract Medium (MSE) were performed with different pH regimes. The antimicrobial activity of the cell-free extracts against Pantoea agglomerans VTT E-90396 and Fusarium avenaceum VTT D-80147 was assessed with automated turbidometry. SIGNIFICANCE AND IMPACT OF THE STUDY: When compared with MRS, the MSE medium cultures produced equal growth yields of Lact. plantarum VTT E-79098 and enhanced antimicrobial potential against the Gram-negative bacterium P. agglomerans and a Fusarium fungus. The MSE medium can be used as a low-cost alternative to MRS for producing high cell yields and good antimicrobial activity of Lact. plantarum.


Assuntos
Grão Comestível/química , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/metabolismo , Extratos Vegetais/química , Antibacterianos/farmacologia , Antifúngicos/farmacologia , Biotecnologia/métodos , Contagem de Colônia Microbiana , Meios de Cultura/química , Meios de Cultivo Condicionados/farmacologia , Fusarium/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana , Pantoea/efeitos dos fármacos
14.
Appl Microbiol Biotechnol ; 63(6): 659-65, 2004 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12910326

RESUMO

The production of nisin, biomass and lactic acid in pH-controlled and uncontrolled batch fermentation and batch fermentation (pH 5.5) with continuous removal of nisin was examined in the parent strain Lactococcus lactis N8 and LAC48. Strain LAC48 in batch fermentor (pH not controlled) gave a maximum nisin concentration of 2.5 x 10(6) IU g dcw(-1). The nisin concentration remained high (2.0 x 10(6) IU g dcw(-1)) after the logarithmic growth phase (10-22 h), whereas nisin production of strain N8 decreased after the logarithmic growth phase. The maximum nisin production of strain LAC48 was not directly related to the biomass formation and not associated with growth. In order to study end product inhibition in nisin production, a system was built for adsorption of nisin during fermentation. The adsorbent Amberlite XAD-4 was found to have an effective binding capacity for nisin. Cells of LAC48 and N8 compensated for the removal of nisin, indicating that nisin production also occurs in the stationary phase.


Assuntos
Lactococcus lactis/crescimento & desenvolvimento , Lactococcus lactis/metabolismo , Nisina/biossíntese , Adsorção , Antibacterianos/biossíntese , Antibacterianos/química , Biomassa , Reatores Biológicos , Biotecnologia/métodos , Fermentação , Glucose/metabolismo , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Nisina/química , Poliestirenos/química , Polivinil/química
15.
Otolaryngol Head Neck Surg ; 129(5): 577-81, 2003 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-14595282

RESUMO

OBJECTIVE: In our previous study, we reported that both uvulopalatopharyngoplasty (UPPP) and tonsillectomy are associated with unacceptable intense pain during the first 24 hours after surgery. To investigate the pain progression at home, we followed the same 53 patients for 14 days after surgery. Twenty-two patients underwent tonsillectomy and 31 patients underwent UPPP. STUDY DESIGN: A prospective, longitudinal, parallel-group study was conducted. METHODS: Beginning on the first postoperative day, the patients were allowed to use 50 mg ketoprofen capsules at a dosage of 5 mg/kg/24 hr. The patients evaluated their own pain four times a day for the first 5 days. Two weeks after the surgery during a follow-up visit patients reported the present pain; and a total consumption of analgesics, and all adverse events for 14 days. RESULTS: Patients in both study groups had significant pain, especially in the morning, for the first week after surgery. In half of the patients, severe pain interfered with eating and sleeping, and in one third of the patients, the pain lasted 2 weeks or longer. The need for ketoprofen was significantly higher after UPPP than that after tonsillectomy (P = 0.001). One patient after tonsillectomy and 3 patients after UPPP needed electrocautery to stop secondary bleeding. CONCLUSIONS: Both UPPP and tonsillectomy are associated with intense postoperative pain. Standard ketoprofen capsules are too short-acting to ensure undisturbed sleep.


Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Cetoprofeno/uso terapêutico , Procedimentos Cirúrgicos Otorrinolaringológicos/métodos , Dor Pós-Operatória/tratamento farmacológico , Palato Mole/cirurgia , Faringe/cirurgia , Úvula/cirurgia , Adulto , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Medição da Dor , Dor Pós-Operatória/diagnóstico , Estudos Prospectivos , Índice de Gravidade de Doença , Tonsilectomia/métodos
16.
Vet Microbiol ; 92(3): 289-93, 2003 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-12523990

RESUMO

Cases of canine distemper (CD) related to vaccination of exotic carnivores extend over three decades and have been described in at least nine different species. Our report describes a case of acute CD in a European mink, Mustela lutreola, vaccinated with live attenuated CD vaccine licensed for use in fur-farmed mink. The male mink died of an acute grey matter disease with an unusually long incubation period. A female vaccinated at the same time showed no obvious signs of illness. The diagnosis was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) and by subsequent sequencing of the PCR products. The sequenced products of the virus isolated from the mink and of the vaccine batch showed 100% identity. This is the first report in which molecular methods were used to confirm that the disease was caused by the vaccine strain. Based on our findings, it is clearly evident that current CD vaccines cannot be safely used in exotic species.


Assuntos
Vírus da Cinomose Canina/imunologia , Cinomose/etiologia , Vison/virologia , Vacinas Virais/efeitos adversos , Animais , Antígenos Virais/análise , Cinomose/patologia , Evolução Fatal , Feminino , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Masculino , Filogenia , RNA Viral/química , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Análise de Sequência de DNA
17.
Mol Hum Reprod ; 8(5): 502-10, 2002 May.
Artigo em Inglês | MEDLINE | ID: mdl-11994548

RESUMO

We have performed comparative genomic hybridization (CGH) analysis of single blastomeres from human preimplantation embryos of patients undergoing preimplantation genetic diagnosis (PGD) for inherited structural chromosome aberrations and from embryos of IVF couples without known chromosomal aberrations. The aim was to verify the PGD results for the specific translocation, reveal the overall genetic balance in each cell and visualize the degree of mosaicism regarding all the chromosomes within the embryo. We successfully analysed 94 blastomeres from 28 human embryos generated from 13 couples. The single cell CGH could verify most of the unbalanced translocations detected by PGD. Some of the embryos exhibited a mosaic pattern regarding the chromosomes involved in the translocation, and different segregation could be seen within an embryo. In addition to the translocations, we found a high degree of numerical aberrations including monosomies, trisomies and duplications or deletions of parts of chromosomes. All of the embryos (100%) were mosaic, containing more than one chromosomally uniform cell line, or even chaotic with a different chromosomal content in each blastomere.


Assuntos
Aberrações Cromossômicas , Embrião de Mamíferos/fisiologia , Hibridização In Situ/métodos , Mosaicismo , Diagnóstico Pré-Implantação/métodos , Blastocisto/patologia , Blastômeros/citologia , Blastômeros/fisiologia , Feminino , Humanos , Masculino , Translocação Genética
18.
Eur J Biochem ; 268(24): 6498-507, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11737205

RESUMO

There are currently four proteins in family 61 of the glycoside hydrolases, from Trichoderma reesei, Agaricus bisporus, Cryptococcus neoformans and Neurospora crassa. The enzymatic activity of these proteins has not been studied thoroughly. We report here the homologous expression and purification of T. reesei Cel61A [previously named endoglucanase (EG) IV]. The enzyme was expressed in high amounts with a histidine tag on the C-terminus and purified by metal affinity chromatography. This is the first time that a histidine tag has been used as a purification aid in the T. reesei expression system. The enzyme activity was studied on a series of carbohydrate polymers. The only activity exhibited by Cel61A was an endoglucanase activity observed on substrates containing beta-1,4 glycosidic bonds, e.g. carboxymethylcellulose (CMC), hydroxyethylcellulose (HEC) and beta-glucan. The endoglucanase activity on CMC and beta-glucan was determined by viscosity analysis, by measuring the production of reducing ends and by following the degradation of the polymer on a size exclusion chromatography system. The formation of soluble sugars by Cel61A from microcrystalline cellulose (Avicel; Merck), phosphoric acid swollen cellulose (PASC), and CMC were analysed on a HPLC system. Cel61A produced small amounts of oligosaccharides from these substrates. Furthermore, Cel61A showed activity against cellotetraose and cellopentaose. The activity of Cel61A was several orders of magnitude lower compared to Cel7B (previously EG I) of T. reesei on all substrates. One significant difference between Cel61A and Cel7B was that cellotriose was a poor substrate for Cel61A but was readily hydrolysed by Cel7B. The enzyme activity for Cel61A was further studied on a large number of carbohydrate substrates but the enzyme showed no activity towards any of these substrates.


Assuntos
Celulase/metabolismo , Trichoderma/enzimologia , Sequência de Bases , Western Blotting , Celulase/genética , Celulase/isolamento & purificação , Celulose 1,4-beta-Celobiosidase , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Cromatografia por Troca Iônica , Clonagem Molecular , Primers do DNA , Eletroforese em Gel de Poliacrilamida , Peso Molecular , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Especificidade por Substrato
19.
Appl Microbiol Biotechnol ; 57(1-2): 182-5, 2001 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11693917

RESUMO

Biofilm-degrading enzymes are potential agents for slime control in paper machines. In this work, extracellular polysaccharides were produced by bacteria isolated from paper machines and the isolated polysaccharides were used as substrates for the screening of polysaccharide-degrading microbes. Polysaccharide yields of 1.5-3.5 g/l were obtained by ethanol precipitation from cultures of strains of Klebsiella pneumoniae, Bacillus licheniformis and Pseudomonas fluorescens on sucrose medium. Two K. pneumoniae strains apparently produced an identical heteropolysaccharide containing galacturonic acid. Fructose-containing polysaccharides were the main products of B. licheniformis and P. fluorescens. Bacteria capable of hydrolyzing the fructose-containing polymers (levans) appeared to be relatively common among the strains selected for screening. None of the bacteria or mixed cultures screened were able to utilize the Klebsiella heteropolysaccharides.


Assuntos
Bactérias/metabolismo , Papel , Polissacarídeos/metabolismo , Bactérias/classificação , Bactérias/enzimologia , Biodegradação Ambiental , Polissacarídeos/química , Especificidade da Espécie
20.
J Histochem Cytochem ; 49(12): 1487-96, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11724896

RESUMO

Cadherin-based molecular complexes play a major role in cell-cell adhesion. At the adherens junctions the intracellular domain of cadherins specifically interacts with beta-catenin and p120ctn, members of the Armadillo repeat protein family. Differential splicing and utilization of the alternative translation initiation codons lead to many p120ctn isoforms. Two major p120ctn isoforms are expressed in mouse tissues. In this study we used indirect immunofluorescence to demonstrate significant tissue specificity in expression of the p120ctn isoforms. The short isoform is abundant at cell-cell adhesion junctions in epidermis, palatal, and tongue epithelia, in the ducts of excretory glands, bronchiolar epithelium, and in mucosal epithelia of esophagus, forestomach, and small intestine. In contrast, the long isoform, containing an amino terminus highly conserved within the p120ctn subfamily, is expressed at vascular-endothelial cell junctions in blood vessels, at cell-cell junctions in the serosal epithelium lining the internal organs, in choroid plexus of brain, in the pigment epithelium of retina, and in structures such as the outer limiting membrane of retina and intercalated discs of cardiomyocytes. The tissue- and cell type-specific expression of p120ctn isoforms suggests a role for the long p120ctn isoform in cell structures responsible for stable tissue integrity, compared to the role of the short isoform in cell-cell adhesion in the external epithelia with rapid turnover.


Assuntos
Moléculas de Adesão Celular/metabolismo , Fosfoproteínas/metabolismo , Animais , Anticorpos Monoclonais , Western Blotting , Cateninas , Moléculas de Adesão Celular/imunologia , Epitélio/metabolismo , Epitélio/ultraestrutura , Técnica Indireta de Fluorescência para Anticorpo , Junções Intercelulares/metabolismo , Camundongos , Especificidade de Órgãos , Fosfoproteínas/imunologia , Isoformas de Proteínas , delta Catenina
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