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1.
Bone Joint J ; 98-B(11): 1450-1454, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27803219

RESUMO

AIMS: Total hip arthroplasty (THA) has well known subjective benefits, but little is known objectively about the recovery of mobility in the early post-operative period. PATIENTS AND METHODS: A total of 33 patients aged > 60 years who underwent elective primary THA had their activity monitored for 30 days post-operatively using an at-home (Fitbit) ankle accelerometer. Their mean age was 70.7 years (61 to 86); 15 (45.5%) were female. The rate of compliance and the mean level of activity were determined. Comparisons between subgroups based on age, body mass index (BMI), surgical approach, and the destination of the patients when discharged were also performed. RESULTS: The mean compliance over the 30 days was 26.7 days (16 to 30; 89%) of use. The mean number of steps increased from 235 (5 to 1152) to 2563 (87 to 7280) (p < 0.001) between the first and the 30th post-operative day. Age < 70 years and an anterior surgical approach were significantly associated with higher levels of activity (1600 to 2400 (p = 0.016 to 0.031) and 1000 to 1800 (p = 0.017 to 0.037) more steps per day, respectively) between the second and the fourth week post-operatively. There was also a trend towards higher levels of activity in those who were discharged to their home rather than to a nursing facility (a mean of 1500 more steps per day, p = 0.02). BMI greater or less than 30 kg/m2 was not predictive of activity (p = 0.45 to 0.98). CONCLUSION: At-home remote mobility monitoring using existing commercially available technology is feasible in patients who have undergone THA. It showed a clear trend towards increased activity with the passage of time. Additionally, the remote device was able to detect differences in levels of activity clearly between patients in relation to variables of interest including age, BMI, surgical approach, and the destination of the patient at the time of discharge from hospital. Such monitoring may allow for the early identification and targeted intervention in patients who recover slowly. Cite this article: Bone Joint J 2016;98-B:1450-4.


Assuntos
Artroplastia de Quadril/reabilitação , Exercício Físico , Cuidados Pós-Operatórios/métodos , Telemetria/métodos , Acelerometria/métodos , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Artroplastia de Quadril/métodos , Estudos de Viabilidade , Feminino , Serviços Hospitalares de Assistência Domiciliar , Humanos , Masculino , Pessoa de Meia-Idade , Cooperação do Paciente , Projetos Piloto , Estudos Prospectivos , Recuperação de Função Fisiológica
2.
Diabetes ; 47(4): 632-9, 1998 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-9568697

RESUMO

Glucagon-like peptide-1 (GLP-1) acts to control blood glucose via multiple mechanisms, including regulation of insulin and glucagon secretion, gastric emptying, satiety, and peripheral insulin sensitivity. However, the relative importance of these actions for regulation of blood glucose remains unclear. We demonstrate here a gene dosage effect for the incretin action of GLP-1, as heterozygous GLP-1R +/- mice exhibit an abnormal glycemic response to oral glucose challenge in association with reduced circulating levels of glucose-stimulated insulin. In contrast, GLP-1 signaling is not required for normal control of fasting and postabsorptive glucagon levels, and no significant changes were detected in the tissue content of pancreatic and intestinal proglucagon mRNA, glucagon-like immunoreactivity, or GLP-1 in GLP-1R -/- or +/- mice. Despite the demonstration that GLP-1 stimulates proinsulin gene transcription, pancreatic insulin mRNA transcripts were similar in wild-type and GLP-1R -/- mice. Furthermore, despite suggestions that GLP-1 regulates peripheral glucose disposal, whole-body glucose utilization was similar in wild-type and GLP-1R -/- mice under both basal and hyperinsulinemic conditions. These observations demonstrate that of the numerous physiological activities ascribed to GLP-1, only the incretin effect on pancreatic beta-cells appears essential for regulation of glucose homeostasis in vivo.


Assuntos
Glicemia/metabolismo , Glucagon/fisiologia , Homeostase , Fragmentos de Peptídeos/fisiologia , Precursores de Proteínas/fisiologia , Receptores de Glucagon/fisiologia , Transdução de Sinais/fisiologia , Alelos , Animais , Feminino , Glucagon/biossíntese , Glucagon/sangue , Peptídeo 1 Semelhante ao Glucagon , Receptor do Peptídeo Semelhante ao Glucagon 1 , Teste de Tolerância a Glucose , Insulina/biossíntese , Insulina/sangue , Mucosa Intestinal/metabolismo , Masculino , Camundongos , Mutação , Pâncreas/metabolismo , Peptídeos/metabolismo , Proglucagon , Precursores de Proteínas/biossíntese , RNA Mensageiro/biossíntese , Radioimunoensaio , Receptores de Glucagon/genética , Transdução de Sinais/genética
3.
Diagn Mol Pathol ; 2(2): 116-9, 1993 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-8269275

RESUMO

There have been conflicting reports regarding the association of cytomegalovirus (CMV) and recurrent spontaneous abortions. It is difficult to assess the role of CMV in the endometrium by histology alone, since the characteristic cytomegalic virocytes are often scarce or absent in this site. Our purpose was to use the polymerase chain reaction (PCR) to detect cytomegalovirus in gestational tissue of women with recurrent spontaneous abortions. DNA was extracted from 25 samples of paraffin-embedded, formalin-fixed gestational tissue from 21 women with at least three unexplained spontaneous abortions (mean, 3.4). DNA from an unstained paraffin section of each specimen was amplified using nested, multiplex PCR specific for the late antigen and the major immediate early genes of CMV. The assay used has a demonstrated level of sensitivity on the order of 10(-2) virocytes per square centimeter of 4-microM paraffin section. Intact DNA was successfully isolated from 21 specimens in 18 patients. Histologic features of CMV infection were completely absent from these cases, and none of these specimens contained evidence of cytomegalovirus DNA. These findings suggest that CMV infection of gestational tissue is not a common direct cause of recurrent spontaneous abortions.


Assuntos
Aborto Habitual/microbiologia , Citomegalovirus/isolamento & purificação , DNA Viral/genética , Reação em Cadeia da Polimerase , Adulto , Sequência de Bases , Vilosidades Coriônicas/microbiologia , Citomegalovirus/genética , Embrião de Mamíferos/microbiologia , Endométrio/microbiologia , Feminino , Humanos , Dados de Sequência Molecular , Gravidez , Sensibilidade e Especificidade , Fixação de Tecidos
4.
Diagn Mol Pathol ; 3(1): 53-8, 1994 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8162256

RESUMO

The purpose of this study was to develop a simple protocol of nested reamplification polymerase chain reaction (PCR) to detect and characterize diverse mycobacterial species. DNA extracted from 126 pure mycobacterial cultures isolated from clinical specimens was amplified by nested PCR with use of a novel set of oligonucleotide primers specific for the 65-kDa antigen gene of mycobacteria. The PCR products were each digested with three restriction enzymes and electrophoresed on an agarose gel. The observed DNA fragment sizes of the different species with each enzyme were compiled into a simple algorithm. This method can rapidly detect and characterize a wide variety of mycobacterial species, including the most common pathogens Mycobacterium tuberculosis, Mycobacterium avium-intracellulare, and Mycobacterium kansasii, without hybridization to labeled probes. The application of this method to surgical pathology was demonstrated by amplification and identification of atypical mycobacteria, including M. kansasii and Mycobacterium leprae, in formalin-fixed paraffin-embedded tissue. This protocol broadens the diagnostic potential of PCR for rapidly diagnosing mycobacterial infection in clinical samples, particularly in paraffin-embedded tissue sections.


Assuntos
Micobactérias não Tuberculosas/classificação , Micobactérias não Tuberculosas/genética , Técnicas de Tipagem Bacteriana , Sequência de Bases , Humanos , Dados de Sequência Molecular , Mycobacterium tuberculosis/genética , Micobactérias não Tuberculosas/isolamento & purificação , Reação em Cadeia da Polimerase
5.
J Clin Pharmacol ; 29(2): 174-84, 1989 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-2715375

RESUMO

The plasma and milk disposition of flurbiprofen (FB) was assessed in healthy women during the early post-partum period after multiple doses of FB. The results confirmed that a pragmatic study design is an attainable requirement for definitive statements about the excretion of FB in transitional milk. Nine doses of FB (50 mg per dose) were administered during three days. Paired milk and plasma samples were obtained during this period of dosing as well as after the last dose. The plasma data were used to derive an equation, which was then used to simulate cumulative plasma profiles for multiple doses given at unequal time intervals. The observed data corresponded to the simulated cumulative profiles of FB in plasma. The plasma elimination half-life of FB during early lactation was slightly prolonged (mean 4.8 hrs) as compared to reported values for normal adult men. The peak plasma concentrations of FB were comparable to those reported for healthy volunteers. In 10 of 12 women (3-5 days post-partum) the FB concentration in breast milk was less than 0.050 micrograms/ml. In two women the milk concentrations of FB were 0.06, 0.07 and 0.07 micrograms/ml as found in only three samples. We conclude that, on the basis of dose found in milk, FB is safe for women breast feeding their infants in the early post-partum period.


Assuntos
Flurbiprofeno/farmacocinética , Leite Humano/metabolismo , Propionatos/farmacocinética , Adulto , Mama/metabolismo , Cromatografia Líquida de Alta Pressão , Feminino , Flurbiprofeno/sangue , Humanos , Lactação , Período Pós-Parto , Gravidez , Espectrofotometria Ultravioleta
6.
Pharmacol Biochem Behav ; 65(4): 743-50, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10764932

RESUMO

Repeated injections of a subconvulsant dose of pentylenetetrazole (PTZ, 30 mg/kg IP three times weekly for 13 injections) in Wistar and hooded Lister rats resulted in kindled seizures, the extent of which varied between strains. Wistar rats achieved stage 4 of clonic-tonic seizures, whereas hooded Lister rats only reached stage 2 of convulsive waves axially through the body. Rats were killed 10 days after their final injection, and radioligand binding was used to measure the expression of NMDA receptors in cortex and hippocampus using [3H]MK-801 and [3H]L-689,560, the latter binding specifically to the NR1 subunit. [3H]Ro 15-1788 measured expression of GABA(A)-benzodiazepine binding sites containing alpha1, alpha2, alpha3, or alpha5 subunits. Specific analysis of GABA(A) receptors containing the alpha5 subunit, which are preferentially localized in the hippocampus, was assessed with [3H]L-655,708. In the cortex, there was no effect of strain or treatment on the K(D) or B(max) of any of the ligands. Similarly, there was no effect of strain or treatment on hippocampal [3H]L-689,560 or [3H]Ro 15-1788 binding. However, in the hippocampus there was a significant, albeit modest, effect of treatment on the B(max) of [3H]MK-801 binding and the B(max) and K(D) of [3H]L-655,708 binding, i.e., PTZ-treated rats had fewer [3H]MK-801 and [3H]L-655,708 binding sites (NMDA and alpha5-containing GABA(A) receptors, respectively), but, these reductions were significant only in the relatively seizure-insensitive hooded Lister strain. This suggests that the increased susceptibility to kindling in Wistar rats is not directly related to alterations in the expression of NMDA or GABA(A) receptors.


Assuntos
Convulsivantes/farmacologia , Excitação Neurológica/efeitos dos fármacos , Pentilenotetrazol/farmacologia , Receptores de GABA-A/biossíntese , Receptores de N-Metil-D-Aspartato/biossíntese , Aminoquinolinas/farmacologia , Animais , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Maleato de Dizocilpina/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Flumazenil/farmacologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Imidazóis/farmacologia , Masculino , Ensaio Radioligante , Ratos , Ratos Wistar , Receptores de GABA-A/efeitos dos fármacos , Receptores de N-Metil-D-Aspartato/efeitos dos fármacos , Especificidade da Espécie , Fatores de Tempo
7.
J Vet Intern Med ; 8(1): 18-25, 1994.
Artigo em Inglês | MEDLINE | ID: mdl-8176658

RESUMO

Serum erythropoietin (Epo) concentrations were measured by radioimmunoassay (RIA) in normal, polycythemic, and anemic dogs and cats. The serum Epo concentration in normal dogs (n = 25) ranged from 7 to 37 mU/mL (median, 20 mU/mL); and in normal cats (n = 11) ranged from 9 to 38 mU/mL (median, 18 mU/mL). Polycythemic animals (PCV > 55% in dogs, > 45% in cats) were classified as those with primary (polycythemia vera), secondary, or polycythemia of uncertain etiology. Dogs with polycythemia vera (PV, n = 8) had a median serum Epo concentration in the normal range (17 mU/mL); cats with PV (n = 7) also had a median serum Epo concentration that was within the normal range (10 mU/mL). In the category of secondary polycythemias, dogs (n = 7) (median, 30.7 mU/mL) and cats (n = 2) had normal Epo concentrations. The median serum Epo concentration was significantly decreased (P < .05) in dogs with PV compared with dogs with secondary polycythemias. The median serum Epo concentrations in dogs (n = 13) and cats (n = 5) with anemias not due to chronic renal disease were significantly increased (P < .05) compared with normal dogs and cats. In cats with anemias due to chronic renal disease (n = 5) the median serum Epo concentration was not significantly different from normal cats. The measurement of the serum EPO concentration may be useful in assessment of anemia or polycythemia but the overlap of values with the normal range in all groups evaluated limit its diagnostic use.


Assuntos
Anemia/veterinária , Doenças do Gato/sangue , Doenças do Cão/sangue , Eritropoetina/sangue , Policitemia/veterinária , Anemia/sangue , Animais , Gatos , Cães , Feminino , Masculino , Policitemia/sangue , Radioimunoensaio/veterinária
8.
J Vet Intern Med ; 7(5): 303-8, 1993.
Artigo em Inglês | MEDLINE | ID: mdl-8263849

RESUMO

Three cats were diagnosed as hyperthyroid based on clinical signs, historical findings, laboratory abnormalities, and basal serum thyroxine (T4) concentrations, and/or nuclear thyroid scans. Additionally, a presumptive diagnosis of thyroid carcinoma with pulmonary metastasis was made in each cat based on radiographic or scintigraphic evaluation. All three cats had solitary pulmonary nodules 1.5 to 2 cm in diameter on survey thoracic radiographs; one cat also had chylous pleural effusion and pulmonary lobar consolidation. Focal pulmonary accumulation of sodium pertechnetate (99mTcO4-) and/or radioiodine (131I) corresponding to radiographic lesions were seen in all cats. Two cats were treated with single ablative doses (1111 to 1480 MBq) of 131I; the remaining cat was euthanatized. One of the treated cats died 8 days later; the other cat was euthanatized 22 weeks following treatment. Histopathologic examination of tissue obtained at necropsy confirmed metastatic thyroid carcinoma in one cat and bronchogenic adenocarcinoma in two cats. Our findings indicate that increased radionuclide uptake in focal pulmonary lesions and cytologic evaluation of tissue obtained by fine-needle aspiration are not specific for thyroid tissue.


Assuntos
Adenocarcinoma/veterinária , Carcinoma/veterinária , Doenças do Gato/diagnóstico , Hipertireoidismo/veterinária , Neoplasias Pulmonares/veterinária , Neoplasias da Glândula Tireoide/veterinária , Adenocarcinoma/diagnóstico , Adenocarcinoma/radioterapia , Adenocarcinoma/secundário , Animais , Carcinoma/diagnóstico , Carcinoma/radioterapia , Carcinoma/secundário , Doenças do Gato/radioterapia , Gatos , Feminino , Hipertireoidismo/diagnóstico , Radioisótopos do Iodo/uso terapêutico , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/radioterapia , Neoplasias Pulmonares/secundário , Pertecnetato Tc 99m de Sódio , Neoplasias da Glândula Tireoide/diagnóstico , Neoplasias da Glândula Tireoide/radioterapia
9.
J Am Vet Med Assoc ; 202(1): 107-9, 1993 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-8420894

RESUMO

Atrophic glomerulopathy resulting in chronic renal failure was diagnosed in 4 related Rottweilers, each < 1 year old. All 4 dogs had severe azotemia and massive protein-losing nephropathy. Histologically, the glomerular lesion was characterized by mild dilatation of Bowman's space, with glomerular tufts absent or markedly atrophied. The lesion is distinct from the congenital glomerular changes described in Samoyeds or Doberman Pinschers.


Assuntos
Doenças do Cão/genética , Nefropatias/veterinária , Falência Renal Crônica/veterinária , Glomérulos Renais/patologia , Proteinúria/veterinária , Animais , Atrofia , Doenças do Cão/etiologia , Doenças do Cão/patologia , Cães , Feminino , Nefropatias/complicações , Nefropatias/genética , Nefropatias/patologia , Falência Renal Crônica/etiologia , Falência Renal Crônica/patologia , Túbulos Renais/patologia , Masculino , Linhagem , Proteinúria/etiologia , Proteinúria/patologia
11.
Mod Pathol ; 9(1): 32-5, 1996 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8821953

RESUMO

Although Mycobacterium paratuberculosis has been reported by both culture and polymerase chain reaction in unfixed intestinal tissue from patients with Crohn's disease, its pathologic significance is unclear. We used polymerase chain reaction to analyze DNA for the presence of M. paratuberculosis from 27 specimens in 23 patients with Crohn's disease. Seventeen of these cases contained granulomata. DNA from unstained paraffin tissue sections was amplified using nested PCR specific for a 264-base pair fragment of IS900, a multicopy insertion element specific to M. paratuberculosis. The sensitivity of this technique was demonstrated by the amplification of small numbers of cultured organisms in a paraffin cell block in the presence of large amounts of added human DNA. M. paratuberculosis was not identified in any of the specimens from patients with Crohn's disease, nor in any of 11 normal colon sections tested. Therefore, we were unable to confirm a role for M. paratuberculosis in Crohn's disease.


Assuntos
Doença de Crohn/microbiologia , DNA Bacteriano/análise , Mycobacterium avium subsp. paratuberculosis/genética , Doença de Crohn/genética , Humanos , Inclusão em Parafina , Paratuberculose/genética , Paratuberculose/microbiologia , Reação em Cadeia da Polimerase
12.
Mod Pathol ; 5(4): 449-54, 1992 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-1323113

RESUMO

Using a simplified procedure, we have extracted DNA from unstained paraffin sections of needle biopsies of kidney and liver transplants and identified the presence of CMV using the polymerase chain reaction. This method utilizes oligonucleotide primers for two genes shown to be specific for cytomegalovirus (CMV) as well as an internal control gene (hemoglobin) in a single reaction. Utilizing nested PCR amplification with agarose gel electrophoresis, CMV can be detected without radioisotopes to a level of sensitivity equivalent to one one-hundredth of a cytomegalic virocyte per cm2 of a 3-microM paraffin section. This method is applicable to situations where only scarce paraffin-embedded tissue is available.


Assuntos
Citomegalovirus/isolamento & purificação , DNA Viral/genética , Sequência de Bases , Biópsia , Citomegalovirus/genética , Humanos , Rim/microbiologia , Fígado/microbiologia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade
13.
Microbiology (Reading) ; 141 ( Pt 10): 2713-8, 1995 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-7582032

RESUMO

Gel-stabilized two-dimensional gradient plates were used to study the effects of pH, salt concentration and temperature on the conjugal transfer of plasmid RP4 between strains of Escherichia coli and Pseudomonas putida. The combinations of pH and salt concentration that permitted conjugation were mapped as a two-dimensional growth area occupied by transconjugants following conjugation. This conjugation domain was less extensive than the areas that supported growth of the parental strains, and showed evidence for the interactive effects of pH and salt concentration in determination of conditions that permitted conjugation. The size and shape of the conjugation domain was influenced by time, temperature, the identities of the donor and recipient bacteria, and the combination of donor and recipient bacteria.


Assuntos
Conjugação Genética , Escherichia coli/genética , Plasmídeos/genética , Pseudomonas putida/genética , Escherichia coli/crescimento & desenvolvimento , Técnicas de Transferência de Genes , Concentração de Íons de Hidrogênio , Biologia Molecular/instrumentação , Pseudomonas putida/crescimento & desenvolvimento , Cloreto de Sódio/farmacologia , Temperatura , Fatores de Tempo
14.
J Neurochem ; 59(5): 1946-54, 1992 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1328533

RESUMO

An anion-exchange HPLC mass assay was used to characterize Swiss-Webster mouse brain and peripheral tissue inositol(1)phosphate [Ins(1)P]levels. Ins(1)P was identified in all tissues studied but Ins(4)P could be identified only in brain, and then only as a part of a peak containing an additional, unidentified component. As a result, it was not possible to quantify Ins(4)P levels. Following a single subcutaneous dose of lithium (10 mmol/kg), brain Ins(1)P levels were maximally elevated after 6 h (corresponding to peak brain lithium concentrations) and were increased to levels 35- and 20-fold higher than in saline-treated animals in cholinergic agonist (pilocarpine)-stimulated and unstimulated animals, respectively. The ED50 for the lithium-induced accumulation of brain Ins(1)P 6 h after administration was 4-6 mmol/kg. The pilocarpine stimulation of lithium-induced brain Ins(1)P accumulation had an ED50 of 22 mg/kg, with maximal accumulation occurring 120 min after pilocarpine administration. Atropine reduced Ins(1)P levels, in both the absence and the presence of lithium, by 40%, indicating that cholinergic systems contribute a large (40%) component of basal brain phosphatidylinositol (PI) cycle activity. In peripheral tissues, there were lithium-induced accumulations of Ins(1)P in kidney, heart, and liver (but not testes) but these were less than that seen in the brain, suggesting that under basal (and pilocarpine-stimulated) conditions, the brain has a higher turnover of the PI cycle than the various peripheral tissues studied. These data support the hypothesis that lithium exerts its effects in vivo via modulation of the PI cycle.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Química Encefálica/efeitos dos fármacos , Cloretos/farmacologia , Fosfatos de Inositol/metabolismo , Lítio/farmacologia , 5'-Nucleotidase/antagonistas & inibidores , Animais , Atropina/farmacologia , Encéfalo/metabolismo , Cloretos/administração & dosagem , Cloretos/metabolismo , Injeções Subcutâneas , Rim/metabolismo , Lítio/administração & dosagem , Lítio/metabolismo , Cloreto de Lítio , Fígado/metabolismo , Masculino , Camundongos , Miocárdio/metabolismo , Fatores de Tempo
15.
Bull World Health Organ ; 68(2): 171-7, 1990.
Artigo em Inglês | MEDLINE | ID: mdl-1694734

RESUMO

Data from 34 studies of the etiology of childhood diarrhoea were compiled in order to investigate the seasonal patterns of rotavirus gastroenteritis and consider their implications for transmission of the virus. Rotavirus was detected in 11-71% of children with diarrhoea, and the median rate of detection (33%) was independent of the level of economic development or geographical region of the study area, as well as of the method of detection used. While rotavirus infections have been called a winter disease in the temperate zones, we found that their incidence peaked in winter primarily in the Americas and that peaks in the autumn or spring are common in other parts of the world. In the tropics, the seasonality of such infections is less distinct and within 10 degrees latitude (north or south) of the equator, eight of the ten locations exhibited no seasonal trend. Throughout most of the world, rotavirus is present all the year round, which suggests that low-level transmission could maintain the chain of infection. The virus is spread by the faecal-oral route but airborne or droplet transmission has also been postulated. The epidemiology of rotavirus--its seasonality in the cooler months, its universal spread in temperate and tropical zones in developed and less developed settings--more closely resembles that of childhood viruses that are spread by the respiratory route (such as measles) than that of common enteric pathogens that are spread predominantly by the faecal-oral route.


Assuntos
Gastroenterite/epidemiologia , Infecções por Rotavirus/epidemiologia , Estações do Ano , América/epidemiologia , Pré-Escolar , Diarreia/microbiologia , Gastroenterite/etiologia , Humanos , Lactente , Recém-Nascido , Rotavirus/isolamento & purificação , Infecções por Rotavirus/transmissão , Clima Tropical
16.
Microbiology (Reading) ; 142 ( Pt 12): 3505-13, 1996 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9004513

RESUMO

Haemophilus influenzae acquires iron from the iron-transporting glycoprotein transferrin via a receptor-mediated process. This involves two outer-membrane transferrin-binding proteins (Tbps) termed Tbp1 and Tbp2 which show considerable preference for the human form of transferrin. Since the Tbps are attracting considerable attention as potential vaccine components, we used transferrin affinity chromatography to examine their conservation amongst 28 H. influenzae type b strains belonging to different outer-membrane-protein subtypes as well as six non-typable strains. Whole cells of all type b and non-typable strains examined bound human transferrin; whilst most strains possessed a Tbp1 of approximately 105 kDa, the molecular mass of Tbp2 varied from 79 to 94 kDa. Antisera raised against affinity-purified native H. influenzae Tbp1/Tbp2 receptor complex cross-reacted on Western blots with the respective Tbps of all the Haemophilus strains examined. When used to probe Neisseria meningitidis Tbps, sera from each of four mice immunized with the Haemophilus Tbp1/2 complex recognized the 68 kDa Tbp2 of N. meningitidis strain B16B6 but not the 78 kDa Tbp2 of N. meningitidis strain 70942. Serum from one mouse also reacted weakly with Tbp1 of strain B16B6. Apart from a weak reaction with the Tbp2 of a serotype 5 strain, this mouse antiserum failed to recognize the Tbps of the porcine pathogen A. pleuropneumoniae. However, a monospecific polyclonal antiserum raised against the denatured Tbp2 of Neisseria meningitidis B16B6 recognized the Tbps of all Haemophilus and Actinobacillus strains examined. Since H. influenzae forms part of the natural flora of the upper respiratory tract, human sera were screened for the presence of antibodies to the Tbps. Sera from healthy adults contained antibodies which recognized both Tbp1 and Tbp2 from H. influenzae but not N. meningitidis. Convalescent sera from meningococcal meningitis patients contained antibodies which, on Western blots, recognized the Tbps2s of both pathogens. These data demonstrate the existence of shared epitopes on the Tbps of H. influenzae, N. meningitidis and A. pleuropneumoniae despite their transferrin species specificity.


Assuntos
Actinobacillus pleuropneumoniae/classificação , Proteínas de Transporte/química , Proteínas de Transporte/imunologia , Haemophilus influenzae/classificação , Neisseria meningitidis/classificação , Actinobacillus pleuropneumoniae/crescimento & desenvolvimento , Actinobacillus pleuropneumoniae/isolamento & purificação , Sequência de Aminoácidos , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/imunologia , Western Blotting , Proteínas de Transporte/isolamento & purificação , Cromatografia de Afinidade , Sequência Conservada , Reações Cruzadas , Haemophilus influenzae/crescimento & desenvolvimento , Haemophilus influenzae/isolamento & purificação , Humanos , Proteínas de Ligação ao Ferro , Camundongos , Dados de Sequência Molecular , Neisseria meningitidis/crescimento & desenvolvimento , Neisseria meningitidis/isolamento & purificação , Proteínas de Ligação a Transferrina
17.
Am J Physiol ; 268(6 Pt 2): H2569-73, 1995 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-7611507

RESUMO

The purpose of this investigation was to determine if exogenous estrogen could attenuate the ventricular arrhythmias caused by myocardial ischemia and reperfusion. Conjugated equine estrogen, administered as an intravenous bolus injection (100 micrograms) to anesthetized, instrumented beagles of both genders, significantly attenuated the incidence of ventricular arrhythmias during a 20-min period of ischemia (2 +/- 1 vs. 19 +/- 16% ectopy) and in the first 5 min of reperfusion (15 +/- 9 vs. 69 +/- 20% ectopy). By 15-20 min of ischemia, ventricular salvos and nonsustained ventricular tachycardia were frequently observed in nontreated dogs. One dog in this group fibrillated during ischemia. In contrast, estrogen-treated dogs exhibited only an occasional ventricular premature beat during the same period of ischemia. When compared with baseline values, the percent ectopy during ischemia in estrogen-treated dogs was insignificant. During reperfusion, nontreated dogs displayed severe, life-threatening arrhythmias such as sustained ventricular tachycardia. In two of these dogs ventricular tachycardia deteriorated to ventricular fibrillation. In comparison, estrogen-treated dogs displayed only innocuous ventricular arrhythmias during reperfusion, i.e., ventricular premature beats, ventricular salvos, and ventricular bigeminy. In addition to the effect of estrogen on arrhythmias, there was a gradual increase in coronary blood flow on reperfusion in estrogen-treated dogs. This effect of estrogen was preceded by a significantly higher coronary perfusion pressure during ischemia (31 +/- 2 vs. 18 +/- 4 mmHg, P < 0.05). In conclusion, our findings suggest that antiarrhythmic effects of estrogen treatment might stabilize ventricular rhythmicity during ischemia and reperfusion.


Assuntos
Arritmias Cardíacas/fisiopatologia , Estrogênios Conjugados (USP)/farmacologia , Hemodinâmica/efeitos dos fármacos , Isquemia Miocárdica/fisiopatologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Disfunção Ventricular/fisiopatologia , Fibrilação Ventricular/fisiopatologia , Animais , Arritmias Cardíacas/etiologia , Pressão Sanguínea/efeitos dos fármacos , Circulação Coronária/efeitos dos fármacos , Cães , Feminino , Frequência Cardíaca/efeitos dos fármacos , Hemodinâmica/fisiologia , Masculino , Análise Multivariada , Taquicardia/etiologia , Taquicardia/fisiopatologia , Disfunção Ventricular/etiologia , Fibrilação Ventricular/etiologia
18.
J Neurochem ; 60(2): 652-8, 1993 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8380439

RESUMO

We have previously described the synthesis of bisphosphonate-containing inhibitors of inositol monophosphatase. In the present study, a more detailed examination of the in vitro and in vivo properties of one of these compounds, L-690,330, is described. L-690,330 is a competitive inhibitor of inositol monophosphatase with a Ki, depending on the source of IMPase, of between 0.2 and 2 microM. Although approximately 1,000-fold more potent in vitro than lithium, in muscarinic ml receptor-transfected Chinese hamster ovary cells prelabelled with [3H]inositol, L-690,330 only produced 40% of the accumulation of [3H]inositol monophosphates achieved by lithium at the same concentration (10 mM), suggesting that the ability of L-690,330 to cross the cell membrane is limited. Nevertheless, under conditions of cholinergic stimulation (100 mg/kg of pilocarpine s.c.), high doses of L-690,330 were able to increase brain inositol(l)phosphate levels in vivo to three- to fourfold control levels. This effect was dose dependent (ED50 = 0.3 mmol/kg s.c.) and was maximal after 1 h. In peripheral tissues, the effects of L-690,330 on inositol(l)phosphate levels mimicked those of lithium both qualitatively and quantitatively. However, in the brain, the effects of L-690,330 were much less than seen with lithium, consistent with the blood-brain barrier restricting access of the polar L-690,330 into the CNS, thereby further limiting entry of compound into cells in the brain. In the future, it may be possible to develop prodrugs of this compound, which circumvent many of the cell permeability problems inherent in bisphosphonate compounds.


Assuntos
Encéfalo/enzimologia , Difosfonatos/farmacologia , Lobo Frontal/enzimologia , Organofosfonatos/farmacologia , Monoéster Fosfórico Hidrolases/antagonistas & inibidores , Animais , Células CHO , Bovinos , Cricetinae , Relação Dose-Resposta a Droga , Humanos , Inositol/metabolismo , Fosfatos de Inositol/metabolismo , Rim/enzimologia , Cinética , Lítio/farmacologia , Fígado/enzimologia , Masculino , Camundongos , Ratos , Proteínas Recombinantes/antagonistas & inibidores , Testículo/enzimologia
19.
Vaccine ; 14(6): 469-77, 1996 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-8782342

RESUMO

We investigated the potential of a herpesvirus of turkey (HVT)-based recombinant virus (rHVT) as an in ovo vaccine to protect specific-pathogen-free chickens against Newcastle disease (ND) and Marek's disease (MD). The rHVT, designed to express fusion (F) and hemagglutinin-neuraminidase (HN) glycoproteins of the lentogenic Hitchner B1 strain of ND virus (NDV), as well as glycoproteins A and B of the GA strain of serotype 1 MD virus (MDV) was efficacious in protecting chickens against ND and MD. No adverse effects on hatchability or the survival of chickens were observed following in ovo vaccination with rHVT. A single administration at embryonation day 18 (ED18) or at hatch protected chickens against challenge-exposures with virulent MDV strain RB-1B and velogenic NDV strain GB-Texas (NDV-GB-TX). Vaccinated chickens developed antibodies against both viruses as detected by serological tests, namely, hemagglutination inhibition, virus neutralization and western immunoblotting for NDV, and immunofluorescence and radioimmunoprecipitation assays for MDV. PCR analysis showed that in ovo vaccination with rHVT resulted in a persistent infection leading to systemic immunity against ND for up to 8 weeks of age, the longest period of time tested in this study. However, virus isolation tests indicated that rHVT-vaccinated chickens were only partially protected from the replication of NDV-GB-TX in the trachea. The results of the study indicate that rHVT is safe for both ED18 and posthatch vaccination for ND and MD, and because the vaccine persists, it may induce longer lasting immunity than conventional live NDV vaccines.


Assuntos
Herpesvirus Galináceo 2/imunologia , Doença de Marek/prevenção & controle , Doença de Newcastle/prevenção & controle , Vacinas Sintéticas/imunologia , Vacinas Virais/imunologia , Animais , Embrião de Galinha , Galinhas , Herpesvirus Galináceo 2/genética , Doença de Newcastle/imunologia , Vírus da Doença de Newcastle/imunologia , Vacinas Sintéticas/efeitos adversos , Vacinas Virais/efeitos adversos , Vacinas Virais/genética
20.
J Neurochem ; 66(6): 2239-47, 1996 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-8632144

RESUMO

Transfection of mouse L(tk-) cells with human N-methyl-D-aspartate (NMDA) receptor subunit cDNAs under the control of a dexamethasone-inducible promoter has been used to generate two stable cell lines expressing NR1a/NR2A receptors and a stable cell line expressing NR1a/NR2B receptors. The cell lines have been characterised by northern and western blot analyses, and the pharmacology of the recombinant receptors determined by radioligand binding techniques. Pharmacological differences were identified between the two NMDA receptor subtypes. The glutamate site antagonist D, L-(epsilon)-2-[3H]amino-4-propyl-5-phosphono-3-pentanoic acid ([3H]CGP 39653) had high affinity for NR1a/NR2A receptors (KD = 3.93 nM) but did not bind to NR1a/NR2B receptors. Glycine site agonists showed a 2.6-5.4-fold higher affinity for NR1a/NR2B receptors. Data from radioligand binding studies indicated that one of the cell lines, NR1a/NR2A-I, expressed a stoichiometric excess of the NR1a subunit, which may exist as homomeric assemblies. This observation has implications when interpreting data from pharmacological analysis of recombinant receptors, as well as understanding the assembly and control of expression of native NMDA receptors.


Assuntos
Linhagem Celular/fisiologia , Receptores de N-Metil-D-Aspartato/genética , 2-Amino-5-fosfonovalerato/análogos & derivados , 2-Amino-5-fosfonovalerato/metabolismo , 2-Amino-5-fosfonovalerato/farmacologia , Aminoquinolinas/metabolismo , Aminoquinolinas/farmacologia , Animais , Sítios de Ligação/fisiologia , Northern Blotting , Maleato de Dizocilpina/metabolismo , Maleato de Dizocilpina/farmacologia , Antagonistas de Aminoácidos Excitatórios/metabolismo , Antagonistas de Aminoácidos Excitatórios/farmacologia , Expressão Gênica/fisiologia , Glutamina/metabolismo , Glutamina/farmacologia , Glicina/metabolismo , Glicina/farmacologia , Humanos , Camundongos , Ensaio Radioligante , Ratos , Receptores de N-Metil-D-Aspartato/agonistas , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Trítio/metabolismo
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