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1.
Australas Phys Eng Sci Med ; 29(2): 203-5, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16845926

RESUMO

We have a developed a novel and practical method of imaging the cornea under ultraviolet (UV) light using a digital medium. Wavelengths of 336-371 nm were used to illuminate the cornea. Images were recorded using a UV Nikkor lens and a digital charged coupled device (CCD). Images obtained showed ferritin lines not visible under white light. This study concluded that 336-371 nm is comparable to shorter wavelengths for the imaging of ferritin in the cornea and that a digital image capture system was comparable to that of film.


Assuntos
Córnea/anatomia & histologia , Aumento da Imagem/instrumentação , Oftalmoscópios , Oftalmoscopia/métodos , Fotografação/instrumentação , Processamento de Sinais Assistido por Computador/instrumentação , Raios Ultravioleta , Desenho de Equipamento , Análise de Falha de Equipamento , Humanos , Aumento da Imagem/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
2.
Br J Ophthalmol ; 90(5): 586-92, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16622088

RESUMO

BACKGROUND: Despite the importance of optical quality of an intraocular lens (IOL) on visual outcomes following cataract surgery, objective data on their optical quality are not readily available, and manufacturing standards are industry regulated. The star test is a classic test of optical quality based on examination of the Airy disc and expanded diffraction rings of a point source of light, used mainly for telescope and microscope objectives. METHODS: A physical model eye cell allowed star testing of IOLs under conditions similar to the optical environment in which they operate. 18 IOLs were tested and results compared to actual images produced by these lenses in the model eye cell. Quantitative measures of star testing performance were developed. RESULTS: The optical performance of the IOLs varied, some performing very poorly. Most lenses (13/17) performed better in reverse orientation, while aberrations induced by the haptics of foldable IOLs were also detected. There was excellent correlation between actual images formed and star testing parameters. CONCLUSION: Star testing IOLs was a novel biomedical application of a centuries old, inexpensive method. A concerning variation of optical quality was found, suggesting IOL optical performance data should be more readily available. Independent, authority mandated IOL optical quality standards should be developed, and results readily available to ophthalmologists.


Assuntos
Lentes Intraoculares/normas , Teste de Materiais , Óptica e Fotônica , Resinas Acrílicas , Sensibilidades de Contraste , Humanos , Modelos Anatômicos , Oftalmologia/métodos , Optometria/normas , Fotografação , Polimetil Metacrilato , Desenho de Prótese , Refração Ocular , Refratometria , Silicones , Acuidade Visual
3.
Thorax ; 61(1): 75-9, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16227331

RESUMO

BACKGROUND: Lower respiratory tract infections (LRTI) are a common reason for consulting general practitioners (GPs). In most cases the aetiology is unknown, yet most result in an antibiotic prescription. The aetiology of LRTI was investigated in a prospective controlled study. METHODS: Eighty adults presenting to GPs with acute LRTI were recruited together with 49 controls over 12 months. Throat swabs, nasal aspirates (patients and controls), and sputum (patients) were obtained and polymerase chain reaction (PCR) and reverse transcriptase polymerase chain reaction (RT-PCR) assays were used to detect Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydia pneumoniae, Legionella pneumophila, influenza viruses (AH1, AH3 and B), parainfluenza viruses 1-3, coronaviruses, respiratory syncytial virus, adenoviruses, rhinoviruses, and enteroviruses. Standard sputum bacteriology was also performed. Outcome was recorded at a follow up visit. RESULTS: Potential pathogens were identified in 55 patients with LRTI (69%) and seven controls (14%; p<0.0001). The identification rate was 63% (viruses) and 26% (bacteria) for patients and 12% (p<0.0001) and 6% (p = 0.013), respectively, for controls. The most common organisms identified in the patients were rhinoviruses (33%), influenza viruses (24%), and Streptococcus pneumoniae (19%) compared with 2% (p<0.001), 6% (p = 0.013), and 4% (p = 0.034), respectively, in controls. Multiple pathogens were identified in 18 of the 80 LRTI patients (22.5%) and in two of the 49 controls (4%; p = 0.011). Atypical organisms were rarely identified. Cases with bacterial aetiology were clinically indistinguishable from those with viral aetiology. CONCLUSION: Patients presenting to GPs with acute adult LRTI predominantly have a viral illness which is most commonly caused by rhinoviruses and influenza viruses.


Assuntos
Infecções Bacterianas/complicações , Infecções Respiratórias/microbiologia , Viroses/complicações , Doença Aguda , Adulto , Idoso , Antibacterianos/uso terapêutico , Infecções Bacterianas/tratamento farmacológico , Estudos de Casos e Controles , Infecções Comunitárias Adquiridas/microbiologia , Infecções Comunitárias Adquiridas/virologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infecções Respiratórias/tratamento farmacológico , Infecções Respiratórias/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Escarro/microbiologia , Viroses/tratamento farmacológico
4.
Eye (Lond) ; 19(6): 683-5, 2005 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15192693

RESUMO

AIMS: Pressure phosphene tonometry is said to assess intraocular pressure by inducing a pressure phosphene. This study compared the results of this relatively new technique with Goldmann applanation tonometry. METHODS: A total of 100 patients (196 readings) in a general ophthalmology clinic at Dunedin Hospital who consented to take part in this study were randomised to receive by different examiners either pressure phosphene tonometry by a Proview eye pressure monitor (Bausch & Lomb Inc., Tampa, FL, USA) or Goldmann tonometry first. There was no communication between the examiners regarding results. RESULTS: Of the 196 attempted readings, pressure phosphene tonometer readings were only able to be obtained for 136 eyes (69%) compared to all 196 (100%) eyes with the Goldmann tonometer. The mean IOPs were 18.5 mmHg using the pressure phosphene tonometer and 16.0 mmHg using the Goldmann tonometer. The mean difference was +2.43 mmHg (95% confidence interval: 10.37 mmHg below to 15.22 mmHg above Goldmann readings). CONCLUSION: This study found that 31% of patients could not perceive a pressure phosphene using the Proview eye pressure monitor. Data obtained from those who could perceive the phosphene indicated that large discrepancies between pressure phosphene tonometry and Goldmann tonometry were common.


Assuntos
Fosfenos , Tonometria Ocular/métodos , Percepção Visual , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Avaliação como Assunto , Glaucoma/diagnóstico , Humanos , Pressão Intraocular , Pessoa de Meia-Idade , Pressão , Reprodutibilidade dos Testes
5.
Am J Respir Crit Care Med ; 164(9): 1618-23, 2001 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-11719299

RESUMO

The effects of respiratory viral infection on the time course of chronic obstructive pulmonary disease (COPD) exacerbation were examined by monitoring changes in systemic inflammatory markers in stable COPD and at exacerbation. Eighty-three patients with COPD (mean [SD] age, 66.6 [7.1] yr, FEV(1), 1.06 [0.61] L) recorded daily peak expiratory flow rate and any increases in respiratory symptoms. Nasal samples and blood were taken for respiratory virus detection by culture, polymerase chain reaction, and serology, and plasma fibrinogen and serum interleukin-6 (IL-6) were determined at stable baseline and exacerbation. Sixty-four percent of exacerbations were associated with a cold occurring up to 18 d before exacerbation. Seventy-seven viruses (39 [58.2%] rhinoviruses) were detected in 66 (39.2%) of 168 COPD exacerbations in 53 (64%) patients. Viral exacerbations were associated with frequent exacerbators, colds with increased dyspnea, a higher total symptom count at presentation, a longer median symptom recovery period of 13 d, and a tendency toward higher plasma fibrinogen and serum IL-6 levels. Non-respiratory syncytial virus (RSV) respiratory viruses were detected in 11 (16%), and RSV in 16 (23.5%), of 68 stable COPD patients, with RSV detection associated with higher inflammatory marker levels. Respiratory virus infections are associated with more severe and frequent exacerbations, and may cause chronic infection in COPD. Prevention and early treatment of viral infections may lead to a decreased exacerbation frequency and morbidity associated with COPD.


Assuntos
Doença Pulmonar Obstrutiva Crônica/virologia , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Viroses/epidemiologia , Idoso , Resfriado Comum/epidemiologia , Feminino , Fibrinogênio/metabolismo , Humanos , Inflamação/sangue , Interleucina-6/sangue , Londres/epidemiologia , Masculino , Prontuários Médicos , Reação em Cadeia da Polimerase/métodos , Estudos Prospectivos , Doença Pulmonar Obstrutiva Crônica/imunologia , Mecânica Respiratória , Infecções por Vírus Respiratório Sincicial/epidemiologia , Infecções Respiratórias/diagnóstico , Risco , Estatísticas não Paramétricas , Viroses/diagnóstico
6.
J Biol Chem ; 276(20): 17172-80, 2001 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-11278563

RESUMO

Tankyrase is an ankyrin repeat-containing poly(ADP-ribose) polymerase originally isolated as a binding partner for the telomeric protein TRF1, but recently identified as a mitogen-activated protein kinase substrate implicated in regulation of Golgi vesicle trafficking. In this study, a novel human tankyrase, designated tankyrase 2, was isolated in a yeast two-hybrid screen as a binding partner for the Src homology 2 domain-containing adaptor protein Grb14. Tankyrase 2 is a 130-kDa protein, which lacks the N-terminal histidine/proline/serine-rich region of tankyrase, but contains a corresponding ankyrin repeat region, sterile alpha motif module, and poly(ADP-ribose) polymerase homology domain. The TANKYRASE 2 gene localizes to chromosome 10q23.2 and is widely expressed, with mRNA transcripts particularly abundant in skeletal muscle and placenta. Upon subcellular fractionation, both Grb14 and tankyrase 2 associate with the low density microsome fraction, and association of these proteins in vivo can be detected by co-immunoprecipitation analysis. Deletion analyses implicate the N-terminal 110 amino acids of Grb14 and ankyrin repeats 10-19 of tankyrase 2 in mediating this interaction. This study supports a role for the tankyrases in cytoplasmic signal transduction pathways and suggests that vesicle trafficking may be involved in the subcellular localization or signaling function of Grb14.


Assuntos
Cromossomos Humanos Par 10 , Poli(ADP-Ribose) Polimerases/genética , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas/metabolismo , Tanquirases , Proteínas Adaptadoras de Transdução de Sinal , Sequência de Aminoácidos , Sítios de Ligação , Linhagem Celular , Cromatografia de Afinidade , Mapeamento Cromossômico , Clonagem Molecular , Biblioteca Gênica , Glutationa Transferase/metabolismo , Humanos , Hibridização in Situ Fluorescente , Dados de Sequência Molecular , Poli(ADP-Ribose) Polimerases/química , Proteínas/química , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/metabolismo , Saccharomyces cerevisiae/genética , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos
7.
Clin Exp Ophthalmol ; 29(6): 416-25, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11778814

RESUMO

AIM: To determine what New Zealand ophthalmologists, general practitioners and optometrists consider important ophthalmic topic areas requiring emphasis in the medical undergraduate curriculum. METHOD: A total of 793 questionnaires related to the content and teaching of undergraduate ophthalmology were sent to ophthalmologists, general practitioners and optometrists. Results were analysed separately for the three respondent groups and as a whole. RESULTS: Four hundred and fourteen questionnaires were returned (52% return rate). Overall responses of the three participant groups were similar and agreed favourably with the current curriculum. The ability to measure visual acuity (97%) and pupillary reflexes (93%), perform ophthalmoscopy (92%), and assess visual fields (68%) were regarded as 'important or essential' by the majority of respondents. Only 53% of respondents consider the ability to diagnose chronic open angle glaucoma as important. The respondents stressed the importance of the diagnosis of predominantly anterior segment disease contrasting with the traditional bias towards the teaching of ophthalmoscopy and posterior segment disease. The majority of respondents stressed the importance of graduating medical students being able to treat bacterial and allergic conjunctivitis, styes, blepharitis, corneal abrasion, and corneal and conjunctival foreign bodies, areas present but not normally emphasized in current curricula. CONCLUSION: The findings of this study provided additional data to facilitate curriculum design and illustrated the value of an integrated problem-based learning approach in ophthalmology undergraduate teaching.


Assuntos
Currículo/normas , Educação de Graduação em Medicina/normas , Medicina de Família e Comunidade/educação , Oftalmologia/educação , Optometria/educação , Competência Clínica/normas , Oftalmopatias/diagnóstico , Oftalmopatias/terapia , Humanos , Aprendizagem , Nova Zelândia , Inquéritos e Questionários , Ensino
8.
J Infect Dis ; 181(6): 1875-84, 2000 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10837165

RESUMO

Rhinoviruses are the major cause of the common cold and a trigger of acute asthma exacerbations. Whether these exacerbations result from direct infection of the lower airway or from indirect mechanisms consequent on infection of the upper airway alone is currently unknown. Lower respiratory infection was investigated in vitro by exposing primary human bronchial epithelial cells to rhinoviruses and in vivo after experimental upper respiratory infection of human volunteers. Bronchial infection was confirmed by both approaches. Furthermore, rhinoviruses induced production of interleukin-6, -8, and -16 and RANTES and were cytotoxic to cultured respiratory epithelium. This evidence strongly supports a direct lower respiratory epithelial reaction as the initial event in the induction of rhinovirus-mediated asthma exacerbations. The frequency of infection and the nature of the inflammatory response observed are similar to those of the upper respiratory tract, suggesting that rhinovirus infections may be one of the most important causes of lower in addition to upper respiratory disease.


Assuntos
Brônquios/virologia , Infecções por Picornaviridae/virologia , Rhinovirus/fisiologia , Células Cultivadas , Citocinas/genética , Efeito Citopatogênico Viral , Células Epiteliais/virologia , Humanos , Hibridização In Situ , RNA Mensageiro/análise , RNA Viral/análise , Rhinovirus/isolamento & purificação
9.
J Wildl Dis ; 36(1): 111-23, 2000 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10682752

RESUMO

Changes in hematological and serum biochemistry parameters in female zinc (Zn)-dosed farm-raised mallards (Anas platyrhynchos) fed four different diets were examined. Sixty ducks received an average dose of 0.97 g of Zn in the form of eight, 3.30-mm diameter shot pellets containing 98% Zn and 2% tin, and another 60 ducks were sham-dosed as controls. Fifteen ducks from each of the two dosing groups were assigned to one of four dietary treatments: corn only, corn with soil, commercial duck ration only, or commercial duck ration with soil. Shot-pellet dissolution rates ranged from 7 mg/Zn/day to 27 mg/Zn/day. Regardless of diet, the Zn dose resulted in mortality; incoordination; paralysis and anorexia; decreased body, liver, pancreas, gonad, and gizzard weight; increased kidney weight; and macroscopic lesions. Zn-dosed ducks had a lower mean erythrocyte packed cell volume (PCV), higher mean reticulocyte count, and a greater number of individuals with immature and/or abnormal erythrocytes, than did control mallards. Mean total leucocyte counts were higher in Zn-dosed ducks than in controls. Zn-dosed ducks that had soil available had higher leucocyte counts than those without soil. Zn-dosed ducks were characterized by a marked heterophilia and relative lymphopenia. In Zn-dosed ducks, the mean lymphocyte count was highest in those provided a commercial duck ration, and lowest in those fed corn. In control ducks, the mean lymphocyte count was highest in ducks fed corn, and lowest in those provided soil along with a commercial duck ration. Zn-dosed mallards had higher serum aspartate aminotransferase and amylase levels, and lower alkaline phosphatase activities than control ducks. Serum phosphorus and uric acid concentrations were higher, and calcium, glucose, and total protein levels lower, in Zn-dosed ducks than in control ducks. Diet did affect serum calcium, phosphorus, total protein, and uric acid concentrations. Differences in erythrocyte and leucocyte parameters, serum enzyme activities, and metabolite concentrations were associated with dose and diet effects. Diets high in protein and other organic matter and calcium and phosphorus did not prevent or substantially alleviate Zn toxicosis in farm-raised mallard ducks.


Assuntos
Doenças das Aves/induzido quimicamente , Dieta/veterinária , Patos/sangue , Zinco/toxicidade , Ração Animal , Animais , Doenças das Aves/sangue , Doenças das Aves/patologia , Contagem de Células Sanguíneas/veterinária , Análise Química do Sangue/veterinária , Peso Corporal/efeitos dos fármacos , Estudos de Coortes , Enzimas/sangue , Enzimas/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Hematócrito/veterinária , Testes Hematológicos/veterinária , Leucócitos/efeitos dos fármacos , Distribuição Aleatória
10.
Eur Respir J ; 16(5): 980-5, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11153603

RESUMO

Rhinovirus (RV) colds are associated with asthma exacerbations and experimental infections are commonly used to investigate the mechanisms involved. However, a temporal association between experimental RV infections and falls in peak expiratory flow (PEF) have not been demonstrated. PEF was measured in 22 volunteers (11 normal, five atopic, six atopic asthmatic) who developed RV serotype 16 colds after inoculation. PEF was measured twice daily for 2 weeks prior and 6 weeks after RV infection and episodes of respiratory morbidity based on changes in PEF were defined using validated criteria. Six significant reductions in PEF were temporally related to the RV infections (in two (18%) normal, one (20%) atopic, three (50%) atopic asthmatic subjects, p=0.1) and occurred 4-9 days (median 6) after inoculation. Mean+/-SEM PEF at day 6 was 87.8+/-1.8% of the predicted value in the six subjects with reductions versus 99.4+/-1.4% pred in those without (p=0.01). Symptom scores were significantly different at day 6 in the two groups (10.6+/-1.9 versus 6.8+/-1.0, p=0.03), but no differences were noted in the viral culture scores and changes in nasal albumin. In subjects with significant PEF reduction, the decrease in the provocative concentration causing a 20% fall in the forced expiratory volume in one second (FEV1) (PC20) was 1.7+/-1.3 mg x mL(-1) versus 1.2+/-1.1 mg x mL(-1) in the negative group (p=0.06). The degree of seroconversion to RV was significantly higher in the group with reduced PEF (median change dilutions 8 versus 4, p=0.02). The results of the present study suggest that rhinovirus-associated, respiratory morbidity occurs during experimental infection in some but not all normal and asthmatic subjects and also that experimental colds are a valid model for the study of rhinovirus-associated airway symptoms and asthma exacerbations.


Assuntos
Resfriado Comum/fisiopatologia , Pico do Fluxo Expiratório , Rhinovirus , Adulto , Asma/complicações , Resfriado Comum/complicações , Feminino , Volume Expiratório Forçado , Humanos , Hipersensibilidade/complicações , Masculino , Pessoa de Meia-Idade , Valores de Referência
11.
J Virol Methods ; 82(1): 9-18, 1999 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10507408

RESUMO

Parainfluenza viruses (PIV) are an important cause of respiratory morbidity. Conventional diagnostic methods for detection of PIV are time consuming or lack sensitivity. A multiplex PCR that detects PIV 1-3 was developed using novel primers for PIV viruses 1 and 2 and primers for PIV 3 described previously. Following RNA extraction a single multiplex reverse transcription was undertaken using antisense primers specific for each virus type. This was followed by a 40-cycle multiplex PCR using primers directed towards the haemagglutinin-neuraminidase coding region of each virus type. Products were probed with type-specific fluorescein labelled internal probes and detected by chemiluminescence. Cultured PIV viruses were detectable to a sensitivity of 1 TCID50. The technique was applied to 57 nasal aspirates taken from children presenting with various acute respiratory conditions and analysed previously by culture, immunofluorescence and/or serology. It was possible to detect PIV 1, 2 or 3 in 13/13 samples found previously positive for PIV by tissue culture, 13/15 found previously positive by immunofluorescence and 6/10 that coincided with positive serology. None of the samples found previously positive for other viruses (26) or negative to virus detection (6) were found positive by RT-PCR. It is concluded that this method is as sensitive as combined immunofluorescence and tissue culture for the detection of the PIV viruses 1-3 and should be useful for rapid diagnosis of PIV 1-3 infections.


Assuntos
Crupe/virologia , Vírus da Parainfluenza 1 Humana/isolamento & purificação , Vírus da Parainfluenza 2 Humana/isolamento & purificação , Vírus da Parainfluenza 3 Humana/isolamento & purificação , Infecções por Respirovirus/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Linhagem Celular , Criança , Crupe/patologia , Técnicas de Cultura/métodos , Imunofluorescência , Humanos , Macaca mulatta , Vírus da Parainfluenza 1 Humana/genética , Vírus da Parainfluenza 2 Humana/genética , Vírus da Parainfluenza 3 Humana/genética , RNA Viral/análise , Infecções por Respirovirus/patologia , Sensibilidade e Especificidade
12.
J Virol Methods ; 80(2): 179-85, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10471027

RESUMO

Rhinoviruses are the main cause of the common cold and precipitate the majority of asthma exacerbations. RT-PCR followed by internal probe hybridisation or Southern blotting, or nested PCRs are currently the most sensitive methods for their identification. However, none of the published techniques can differentiate satisfactorily rhinoviruses from other picornaviruses. Examination of the restriction maps of sequenced rhinoviruses, revealed a highly conserved BglI restriction site (GCCnnnnnGGC), located exactly in the middle of the 380-bp amplicon generated with the OL26-OL27 primer pair, which has been used extensively in the past to identify picornaviruses. Such a site was either not present, or positioned differently in other picornaviruses of known sequence. It was, therefore, considered that digestion of rhinovirus amplicons with this enzyme would result in two equal length fragments, generating a single 190-bp band in gel electrophoresis. In contrast, either one undigested 380-bp band or a double-band pattern would appear in amplicons from other picornaviruses. To test this hypothesis, Bgl digestions of OL26-OL27 amplicons from cultured and wild-type rhinoviruses, whose identity was confirmed by acid lability, as well as from echo, polio and coxsackie viruses were carried out. All rhinovirus samples were digested successfully generating single bands. Among the other picornaviruses, only 6.6% presented a single band pattern, while the rest were as predicted from the model. With a sensitivity of 100% and a specificity over 90%, the method described, which is rapid and remarkably easy to perform, can be used to distinguish rhinoviruses from other picornaviruses to a considerable extent.


Assuntos
Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Rhinovirus/isolamento & purificação , Eletroforese em Gel de Ágar , Eletroforese em Gel de Poliacrilamida , Amplificação de Genes/genética , Genes Virais/genética , Genoma Viral , Células HeLa , Humanos , Picornaviridae/genética , Polimorfismo de Fragmento de Restrição , Rhinovirus/classificação , Rhinovirus/genética
13.
J Med Virol ; 58(1): 100-4, 1999 May.
Artigo em Inglês | MEDLINE | ID: mdl-10223554

RESUMO

Rhinoviruses are epidemiologically connected to the majority of acute asthma exacerbations; however, their ability to infect and replicate in the lower airways is disputed. A frequent argument against this possibility involves the temperature preference for rhinovirus replication, generally accepted to be 33 degrees C, the temperature of the nasal passages. However, this argument is based on studies with a single rhinovirus serotype. In this study, differences in temperature preferences were evaluated between several serotypes and relative titers were determined than can be achieved at upper and lower airway temperatures. Rhinovirus serotypes 1b, 2, 7, 9, 14, 16, 41, and 70 were titrated in Ohio-HeLa cell cultures at either 33 degrees C or 37 degrees C. Possible selection by culture temperature was examined by continuous culture at 33 degrees C and 37 degrees C for 2-4 passages and subsequent titration at both temperatures. Finally, nasal aspirate samples derived from patients with wild-type rhinoviral common colds were cultured at 33 degrees C and 37 degrees C and RT-PCR was used to assess rhinovirus replication at each temperature. The majority of the serotypes and wild-type viruses replicated slightly better at 33 degrees C than at 37 degrees C. However, titers achieved after one or more replicative cycles at 37 degrees C were still high enough to initiate infection. Furthermore, in some instances equal or even better replication was observed at 37 degrees C. It is concluded that temperature preferences may vary between rhinoviruses and are not likely to be a prohibitive factor for infection of the lower airways.


Assuntos
Rhinovirus/fisiologia , Replicação Viral/fisiologia , Células HeLa , Humanos , Rhinovirus/genética , Rhinovirus/crescimento & desenvolvimento , Temperatura
14.
J Wildl Dis ; 34(4): 688-97, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9813837

RESUMO

Effects of five lead (Pb), iron (Fe), or bismuth (Bi)/tin (Sn) alloy shot embedded in the breast muscles of game-farm mallards (Anas platyrhynchos) were studied from 28 March 1994 through 27 March 1995. We detected no differences in the mean survival times, mean hematocrits, or mean body weights among the three shot types. Connective tissue encapsulated Pb and Bi/Sn shot but only slight changes occurred in tissues surrounding the shot. Recovered Pb and Bi/Sn shot were essentially unchanged in appearance and weight. A thin zone of "oxide" surrounded Fe shot with a slight inflammatory response and a small amount of scarring adjacent to the embedded shot. Fe shot decreased slightly in weight while embedded. Bacterial infections were absent in all dosed ducks. Mean weights of kidneys, livers, and gonads did not vary by type of shot. Kidneys and livers of Bi-dosed ducks had higher concentrations of Bi than in Pb- and Fe-dosed ducks. Muscle and blood showed no differences in Bi concentrations among doses. We found no histological dose-related effects in kidneys, liver, and gonads from the embedded shot.


Assuntos
Doenças das Aves/induzido quimicamente , Bismuto/toxicidade , Patos , Ferro/toxicidade , Chumbo/toxicidade , Músculos Peitorais/efeitos dos fármacos , Ligas , Animais , Doenças das Aves/mortalidade , Doenças das Aves/patologia , Bismuto/farmacocinética , Peso Corporal/efeitos dos fármacos , Feminino , Gônadas/efeitos dos fármacos , Gônadas/metabolismo , Gônadas/patologia , Ferro/farmacocinética , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Chumbo/farmacocinética , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Músculos Peitorais/metabolismo , Músculos Peitorais/patologia , Intoxicação/mortalidade , Intoxicação/patologia , Intoxicação/veterinária , Distribuição Aleatória
15.
J Virol Methods ; 67(2): 153-60, 1997 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9300380

RESUMO

We describe an in-situ RT-PCR method for the amplification of rhinovirus (RV) in fixed, paraffin-embedded HeLa cells employed as a model for human respiratory epithelium. HeLa cells were infected in-vitro with inocula of rhinovirus-16 ranging from 10(2) to 10(6) 50% tissue culture infective doses (TCID50), incubated for 18 h then fixed and processed into paraffin blocks. Sections of the cell preparation were subjected to standard RT-PCR, in-situ hybridisation (ISH) or in-situ RT-PCR using specific oligonucleotide primers or probes directed against the 5' non-coding region of RV RNA. RT-PCR was found to be capable of detecting RV16 RNA in one 8 microns-thick section of cells infected with the lowest virus titre. ISH using digoxigenin labelled oligonucleotide probes located RV16 signal in the majority of HeLa cells at the highest virus titre, but in few or no cells with the lowest virus titre. In contrast, in-situ RT-PCR detected RV16 in the majority of cells infected with this amount of RV16. There was a slight loss of morphology and fine localisation associated with the in-situ thermal cycling process. However, the sensitivity of in-situ RT-PCR is comparable to standard RT-PCR and greater than ISH for the detection of RV. In-situ RT-PCR has wide applications for sensitive localization of low copy viral and RNA sequences within cells to investigate the role of viruses in a variety of clinical conditions.


Assuntos
Hibridização In Situ/métodos , Inclusão em Parafina , Infecções por Picornaviridae/virologia , Reação em Cadeia da Polimerase/métodos , Rhinovirus/genética , Células HeLa , Humanos , Infecções por Picornaviridae/diagnóstico , Infecções por Picornaviridae/patologia , RNA Viral/química , RNA Viral/isolamento & purificação , DNA Polimerase Dirigida por RNA , Rhinovirus/isolamento & purificação , Sensibilidade e Especificidade
16.
Aust N Z J Ophthalmol ; 25 Suppl 1: S87-90, 1997 May.
Artigo em Inglês | MEDLINE | ID: mdl-9267637

RESUMO

Multifocal flash electroretinograms (ERG) were recorded binocularly (n = 18). Areas were equal or scaled with excentricity. The latter were expected to increase the total amplitude if ganglion cell subcomponents were involved. Amplitudes were intercorrelated and the factor structure was established. Scaling had no influence on amplitudes or on factors. Reliability and sensitivity were high. The second kernel first slice from the nine hexagons across the midline showed differences near the macula but no component increasing in latency with distance from the disc. Thus, multifocal flash ERG have linear spatial summation and no ganglion cell subcomponents.


Assuntos
Eletrorretinografia , Estimulação Luminosa , Células Ganglionares da Retina/fisiologia , Humanos , Sensibilidade e Especificidade
17.
J Biol Chem ; 272(13): 8490-7, 1997 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-9079677

RESUMO

The Src homology 2 (SH2) domain-containing protein Grb7 and the erbB2 receptor tyrosine kinase are overexpressed in a subset of human breast cancers. They also co-immunoprecipitate from cell lysates and associate directly in vitro. Whereas the Grb7 SH2 domain binds strongly to erbB2, the SH2 domain of Grb14, a protein closely related to Grb7, does not. We have investigated the preferred binding site of Grb7 within the erbB2 intracellular domain and the SH2 domain residues that determine the high affinity of Grb7 compared with Grb14 for this site. Phosphopeptide competition and site-directed mutagenesis revealed that Tyr-1139 of erbB2 is the major binding site for the Grb7 SH2 domain, indicating an overlap in binding specificity between the Grb7 and Grb2 SH2 domains. Substituting individual amino acids in the Grb14 SH2 domain with the corresponding residues from Grb7 demonstrated that a Gln to Leu change at the betaD6 position imparted high affinity erbB2 interaction, paralleled by a marked increase in affinity for the Tyr-1139 phosphopeptide. The reverse switch at the betaD6 position abrogated Grb7 binding to erbB2. This residue therefore represents an important determinant of SH2 domain specificity within the Grb7 family.


Assuntos
Proteínas/metabolismo , Receptor ErbB-2/metabolismo , Domínios de Homologia de src , Proteínas Adaptadoras de Transdução de Sinal , Sequência de Aminoácidos , Sítios de Ligação , Ligação Competitiva , Células Cultivadas , Proteína Adaptadora GRB7 , Humanos , Modelos Moleculares , Dados de Sequência Molecular , Proteínas/química , Receptor ErbB-2/química , Alinhamento de Sequência , Relação Estrutura-Atividade , Tirosina/metabolismo
18.
Eur Respir J ; 9(11): 2250-5, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8947068

RESUMO

Experimental viral disease studies in volunteers have clarified many aspects of the pathogenesis of human viral disease. Recently, interest has focused on rhinovirus-associated asthma exacerbations, and new volunteer studies have suggested that airway responsiveness (AR) is enhanced during a cold. For scientific, ethical and safety reasons, it is important to use validated methods for the preparation of a virus inoculum and that the particular virological characteristics and host responses should not be altered. We have prepared a new human rhinovirus (HRV) inoculum using recent guidelines and assessed whether disease characteristics (for example, severity of colds or changes in AR) were retained. Studies were conducted in 25 clinically healthy volunteers using a validated HRV inoculum in the first 17 and a new inoculum in the subsequent eight subjects. Severity of cold symptoms, nasal wash albumin levels and airway responsiveness were measured, and the new inoculum was prepared from nasal washes obtained during the cold. The new inoculum was tested using standard virological and serological techniques, as well as a polymerase chain reaction for Mycoplasma pneumoniae. No contaminating viruses or organisms were detected and the methods suggested were workable. Good clinical colds developed in 20 of the 25 subjects and median symptom scores were similar in the validated and new inoculum groups (18 and 17.5, respectively; p=0.19). All subjects shed virus, and there were no differences noted in viral culture scores, nasal wash albumin and rates of seroconversion in the two groups. Although airway responsiveness increased in both groups (p=0.02 and p=0.05), the degree of change was similar. We have performed experimental rhinovirus infection studies and demonstrated similar clinical disease in two inoculum groups. Amplified airway responsiveness was induced; continuing studies will define the mechanisms and suggest modes of treatment.


Assuntos
Infecções por Picornaviridae/fisiopatologia , Rhinovirus , Adulto , Albuminas/análise , Animais , Anticorpos Antivirais/sangue , Testes de Provocação Brônquica , Humanos , Camundongos , Pessoa de Meia-Idade , Mycoplasma pneumoniae/isolamento & purificação , Líquido da Lavagem Nasal/química , Líquido da Lavagem Nasal/microbiologia , Líquido da Lavagem Nasal/virologia , Infecções por Picornaviridae/virologia , Rhinovirus/imunologia
19.
Am J Respir Crit Care Med ; 154(3 Pt 1): 654-60, 1996 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8810601

RESUMO

We have shown that viruses are associated with 80 to 85% of asthma exacerbations in school-age children in the community. We hypothesize that viral infections are also associated with severe attacks of asthma precipitating hospital admissions. To investigate this, we conducted a time-trend analysis, comparing the seasonal patterns of respiratory infections and hospital admissions for asthma in adults and children. During a 1-yr study in the Southampton area of the United Kingdom, 108 school-age children monitored upper and lower respiratory symptoms and took peak expiratory flow rate (PEFR) recordings. From children reporting a symptomatic episode or a decrease in PEFR, samples were taken for detection of viruses and atypical bacteria. A total of 232 respiratory viruses and four atypical bacteria were detected. The half-monthly rates of upper respiratory infection were compared with the half-monthly rates for hospital admissions for asthma (International Classification of Diseases [ICD] code 493) for the same time period for the hospitals serving the areas from which the cohort of schoolchildren was drawn. The relationships of upper respiratory infections and hospital admissions for asthma with school attendance were studied. Strong correlations were found between the seasonal patterns of upper respiratory infections and hospital admissions for asthma (r = 0.72; p < 0.0001). This relationship was stronger for pediatric (r = 0.68; p < 0.0001) than for adult admissions (r = 0.53; p < 0.01). Upper respiratory infections and admissions for asthma were more frequent during periods of school attendance (87% of pediatric and 84% of total admissions), than during school holiday periods (p < 0.001). These relationships remained significant when allowance was made for linear trend and seasonal variation using multiple regression analysis (p < 0.01). Not surprisingly, school attendance, because it is a major factor in respiratory virus transmission, was found to be a major confounding variable in children. This study demonstrates that upper respiratory viral infections are strongly associated in time with hospital admissions for asthma in children and adults. Rhinoviruses were the major pathogen implicated, and the majority of viral infections and asthma admissions occurred during school attendance.


Assuntos
Asma/epidemiologia , Hospitalização/estatística & dados numéricos , Infecções Respiratórias/epidemiologia , Adolescente , Adulto , Criança , Feminino , Humanos , Estudos Longitudinais , Masculino , Pico do Fluxo Expiratório , Prevalência , Infecções Respiratórias/virologia , Estações do Ano , Fatores de Tempo , Reino Unido/epidemiologia
20.
J Cataract Refract Surg ; 22(5): 615-9, 1996 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-8784637

RESUMO

PURPOSE: To assess depth of field in phakic and pseudophakic eyes to explain good distance and uncorrected near visual acuity in pseudophakic eyes. SETTING: Department of Ophthalmology, University of Otago Medical School, Dunedin, New Zealand. METHODS: Depth of field was measured in pseudophakic (n = 10) and phakic (n = 10) eyes for both near and distant targets. Test conditions included cycloplegia and a constant pupillary aperture using a soft contact lens with a central artificial pupil diameter of 2.5 mm. RESULTS: There was no statistically significant difference between phakic and pseudophakic eyes for any test. Depth of field for near visual acuity was +/-0.85 diopters (D), but amplitude of legibility was +/-1.94 D. Depth of field for distance visual acuity was between 0.25 and 0.50 D in 85% of eyes. CONCLUSION: In the absence of astigmatism and disease, a pseudophakic eye with -0.75 D of myopia can expect to achieve 20/30 uncorrected distance acuity and read N5 unaided if the pupil is approximately 2.5 mm.


Assuntos
Acomodação Ocular , Extração de Catarata , Percepção de Profundidade , Lentes Intraoculares , Lentes de Contato , Humanos , Cristalino/fisiologia , Erros de Refração/fisiopatologia , Acuidade Visual
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