RESUMO
Creutzfeldt-Jakob disease (CJD) is a rare, fatal neurodegenerative disease that occurs in sporadic, genetic, variant, and iatrogenic forms. The transformation of normal prion protein (PrP(C)) to the abnormal form (PrP(Sc)) is a key step in the pathogenesis of CJD and leads to the accumulation of amyloid and spongiform changes in the brain. The presence of PrP(Sc) in tissue is a surrogate marker for CJD infectivity. Sporadic CJD, whose cause is unknown, is by far the most frequent form with 1-2 cases per million population occurring every year-the genetic forms of CJD are rather rarer. The majority of variant CJD cases have occurred in the United Kingdom, where there have been four reports of transmission of vCJD by blood transfusion. The great majority of iatrogenic transmissions of CJD have resulted from the use of pituitary-derived hormones or dura mater with only a very few cases attributable to neurosurgical instruments or corneal transplants. In the absence of a validated test for CJD infectivity in eye donors, the application of appropriate donor selection criteria and the use of single-use instruments in eye banks are currently the most effective means of reducing the risk of CJD transmission. Onward transmission by reusable ophthalmic surgical instruments has not been reported, but the risk cannot be excluded. Use of appropriate cleaning and disinfection protocols and the ability to identify and quarantine instruments that may have been used on an infected patient are important safeguards.
Assuntos
Transplante de Córnea/efeitos adversos , Síndrome de Creutzfeldt-Jakob/transmissão , Infecção Hospitalar/prevenção & controle , Síndrome de Creutzfeldt-Jakob/epidemiologia , Humanos , Retina/transplante , Instrumentos Cirúrgicos , Reino Unido/epidemiologiaRESUMO
PURPOSE: To describe the incidence and current management of fungal keratitis in the United Kingdom. METHODS: Cases were identified prospectively through the British Ophthalmologic Surveillance Unit (BOSU) from December 2003 to November 2005. Questionnaire data were requested at diagnosis and at 6 months follow-up. Inclusion criteria were a positive culture or microsopic proof from a scraping or biopsy, and a normal residence in the United Kingdom. RESULTS: Data were available on 39 confirmed cases at diagnosis and 34 cases at follow-up. The minimum average annualised incidence was 0.32 (95% CI, 0.24-0.44) cases per million individuals. In 22 cases (56%), only Candida was isolated and 14 of these (63%) had prior ocular surface disease treated with topical steroid. A filamentary fungus infection was more common in male patients (P=0.02), often following trauma, and the differences in risk factors between types of fungal infection was statistically significant (P<0.001). One case had a mixed yeast and filamentary fungus infection. The most frequent initial topical therapies were amphotericin B (38%) or econazole (28%). In addition, oral fluconazole was used in 11 (31%) patients and oral itraconazole in six (15%). At follow-up, the vision in 15 eyes (44%) was <6/60 including three eyes eviscerated. CONCLUSIONS: This study provides data on the minimum incidence of fungal keratitis in the United Kingdom. It provides evidence of frequent delay in diagnosis after presentation to eye departments, inconsistent management, and poor outcome. Issues that can now be addressed.
Assuntos
Infecções Oculares Fúngicas/epidemiologia , Infecções Oculares Fúngicas/terapia , Ceratite/epidemiologia , Ceratite/terapia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antifúngicos/uso terapêutico , Infecções Oculares Fúngicas/microbiologia , Feminino , Humanos , Incidência , Ceratite/microbiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Risco , Reino Unido/epidemiologia , Adulto JovemRESUMO
AIMS: To determine the epithelial proliferative capacity of organ cultured limbal tissue and correlate this with various donor and eye banking factors. METHODS: 24 corneoscleral limbal (CSL) rims left over from penetrating keratoplasty were split in half and set up as in vitro explant cultures. Corneal epithelial proliferative potential (CEPP) was assessed by the number of "cycles" of growth achieved before explants underwent exhaustion and failure to generate an epithelium to subconfluence. The dependence of CEPP on the age of the donor, time of death to enucleation, time of enucleation to organ culture, and time in organ culture in the eye bank was determined. RESULTS: CSL rims were capable of up to four cycles of culture with a wide variation between tissue samples. Of the various factors examined, death to enucleation time was the only statistically significant factor affecting the CEPP (regression coefficient: -0.062 (cycles/hour), CI -0.119 to -0.004, p = 0.037). Time in organ culture had little effect on CEPP. CONCLUSIONS: Preselected organ cultured CSL rims from eye banks may offer a viable alternative tissue source for use in allo-limbal transplantation.
Assuntos
Transplante de Córnea/métodos , Epitélio Corneano/citologia , Bancos de Olhos , Limbo da Córnea/citologia , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Proliferação de Células , Meios de Cultura , Humanos , Pessoa de Meia-Idade , Técnicas de Cultura de Órgãos , Transplante de Células-Tronco , Células-Tronco/citologia , Fatores de Tempo , Preservação de Tecido/métodos , Coleta de Tecidos e ÓrgãosRESUMO
PURPOSE: To describe the strategy used for large-scale ophthalmological monitoring in the clinical development of the novel anticancer agent gefitinib ('Iressa', ZD1839), an epidermal growth factor receptor tyrosine kinase inhibitor, which had demonstrated ocular effects in preclinical animal models. METHODS: In this extensive clinical trial programme, patients in Phase I and II trials underwent frequent and intensive ophthalmological monitoring at baseline and during the trials. Data were reviewed by an external independent Ophthalmology Advisory Board. RESULTS: Ophthalmological data for 221 patients in Phase I trials of gefitinib and 425 patients in Phase II trials revealed no evidence of any consistent or drug-related ophthalmological toxicity. Interestingly, the baseline data revealed that, in an asymptomatic population, transient ophthalmological events are identified during monitoring. CONCLUSIONS: This study reports the methodology and normative data in an ophthalmological screening programme that should prove useful for future studies.
Assuntos
Antineoplásicos/efeitos adversos , Receptores ErbB/antagonistas & inibidores , Oftalmopatias/induzido quimicamente , Inibidores de Proteínas Quinases/efeitos adversos , Quinazolinas/efeitos adversos , Adulto , Idoso , Idoso de 80 Anos ou mais , Monitoramento de Medicamentos , Feminino , Gefitinibe , Humanos , Masculino , Pessoa de Meia-Idade , Transtornos da Visão/induzido quimicamenteRESUMO
PURPOSE: To determine the prevalence and clinical characteristics of external ocular infections caused by methicillin-resistant Staphylococcus aureus (MRSA) in an ophthalmic hospital in the UK. METHODS: A retrospective analysis of the case notes of patients who had culture proven external ocular Staphylococcal infections during a 44-month period was undertaken. RESULTS: There were a total of 548 external eye infections caused by Staphylococcus aureus. Of these, 17 (3%) were MRSA positive. The most common presentation was conjunctivitis seen in six patients. All MRSA isolates were sensitive to chloramphenicol. Ofloxacin resistance was observed in all isolates from patients over the age of 50 years. All patients had an underlying history of either an ocular surface disease, malignancy, or a debilitating medical illness. CONCLUSIONS: MRSA is as yet an infrequent cause of external ocular infections. Patients typically have underlying ocular risk factors and/or are medically debilitated. Different strains infect young and old age groups with characteristic antimicrobial sensitivity. This study highlights the need for more work to establish the role of MRSA commensals and ocular infections.
Assuntos
Infecções Oculares Bacterianas/microbiologia , Resistência a Meticilina , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/efeitos dos fármacos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Conjuntivite Bacteriana/epidemiologia , Conjuntivite Bacteriana/microbiologia , Infecção Hospitalar/microbiologia , Inglaterra/epidemiologia , Infecções Oculares Bacterianas/epidemiologia , Humanos , Ceratite/epidemiologia , Ceratite/microbiologia , Pessoa de Meia-Idade , Prevalência , Estudos Retrospectivos , Infecções Estafilocócicas/epidemiologiaRESUMO
AIMS: To describe the clinical features of two patients with paraproteinaemia and necrobiotic xanthogranulomatosis together with detailed immunohistochemistry of the lesions in one. METHODS: The clinical history and results of biochemical investigations of the patients were retrieved from the files. Immunohistochemistry was used to investigate the expression of macrophage and mast cell markers, amyloid A and P, S-100 protein, and apolipoprotein AI and B in xanthogranulomatous skin lesions from patient 2. In addition, protein A-sepharose chromatography was used to separate serum from patient 2 and apolipoprotein B and the IgG paraprotein were measured in the fractions eluted. RESULTS: Monocytes/macrophages comprised the major cellular component of the lesion, and unusually for xanthomata, areas of collagen necrosis were also seen. Activated mast cells were present at the margins of macrophage clusters and adjacent to areas of collagen necrosis. Serum paraprotein was bound to low density lipoproteins as judged by protein A-sepharose chromatography, and was also located within macrophagic foam cells of the lesion on immunohistochemistry. CONCLUSIONS: These observations demonstrate many features similar to atherosclerosis including collagen necrosis and mast cell activation.
Assuntos
Granuloma/patologia , Transtornos Necrobióticos/patologia , Xantomatose/patologia , Idoso , Feminino , Granuloma/metabolismo , Humanos , Macrófagos/patologia , Pessoa de Meia-Idade , Monócitos/patologia , Transtornos Necrobióticos/metabolismo , Paraproteinemias/metabolismo , Paraproteinemias/patologia , Xantomatose/metabolismoAssuntos
Autoanticorpos/análise , Úlcera da Córnea/complicações , Úlcera da Córnea/imunologia , Granulomatose com Poliangiite/imunologia , Glicoproteínas de Membrana/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Córnea/imunologia , Proteína Ligante Fas , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Our purpose is to examine levels of Fas mRNA expression in blood during human corneal transplant rejection. METHODS: Fas mRNA expression was detected by reverse transcription-PCR in blood from normal controls, corneal recipients at the time of transplantation and during episodes of rejection. RESULTS: Samples taken at the time of a corneal rejection episode showed Fas mRNA levels were significantly lower in these patients than either normal controls (P = 0.017) or corneal transplant recipients not undergoing graft rejection (P = 0.00052). Serial samples from five patients who suffered an episode of rejection showed that the level of Fas mRNA is reduced during the rejection episode and subsequently recovers. CONCLUSIONS: These results indicate low levels of Fas mRNA in blood may have a role in corneal transplant rejection.
Assuntos
Transplante de Córnea/imunologia , Rejeição de Enxerto/sangue , RNA Mensageiro/sangue , Receptor fas/genética , Adulto , Humanos , Pessoa de Meia-IdadeRESUMO
Data relating to 3313 adenovirus isolates from patients in Greater Manchester, UK between 1982 and 1996 were analysed using chi2 tests and 95% confidence intervals. Of the 3098 isolates that were typed, 18.6% were serotype 2, 14.9% serotype 3, 12.1% serotype 1 and 10.9% serotype 41. There was evidence of a seasonal occurrence of serotype 7 (March-August), serotype 2 (January-April), serotype 4 (June-August) and subgenus F (September-November). Children less than 5 years old were the most common group of patients with adenovirus infection (61.3%) compared to 24.2% for adults and only 5.6% for school children (5-15 years). Gastric symptoms were the most common amongst infants (47.6%) followed by respiratory (27.5%) and general symptoms (12.9%). In adults, the overwhelming clinical condition was conjunctivitis (88.9%). Despite the traditional association with adenoviruses, remarkably few cases of pharyngoconjunctival fever were recorded (1.7%).
Assuntos
Infecções por Adenoviridae/epidemiologia , Adenoviridae/classificação , Conjuntivite/etiologia , Adenoviridae/patogenicidade , Infecções por Adenoviridae/diagnóstico , Infecções por Adenoviridae/patologia , Adolescente , Adulto , Fatores Etários , Idoso , Criança , Pré-Escolar , Inglaterra/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , SorotipagemRESUMO
AIM: To report the clinical consequences of contamination of human donor corneas by herpes simplex virus (HSV) in organ culture. METHODS: Two patients without previous history of ocular HSV infection underwent penetrating keratoplasty (PK), one for keratoconus and the other for Fuchs' endothelial dystrophy. One patient suffered primary graft failure while the other developed a persistent epithelial defect, ultimately resulting in graft failure. Viral culture of swabs taken from both corneas during the early postoperative period was undertaken. The failed donor corneas were examined histopathologically by immunohistochemistry (IHC) for HSV-1 antigens, transmission electron microscopy (TEM), and by polymerase chain reaction (PCR) for HSV DNA. Both failed corneas were replaced within 6 weeks of the initial surgery. The records of the fellow donor corneas were also examined for evidence of infection. RESULTS: HSV was cultured from both corneas during the early postoperative period. Histology of both donor corneas demonstrated a thickened corneal stroma with widespread necrosis of keratocytes and loss of endothelial cells. IHC showed keratocytes positive with antibodies to HSV-1 antigens. TEM demonstrated HSV-like viral particles within degenerating keratocytes. PCR performed on the failed corneal grafts was positive for HSV-1 DNA, whereas PCR performed on the excised host corneal buttons was negative in both patients. Records of the fellow donor corneas showed that one cornea was successfully transplanted into another recipient after 18 days in organ culture, whilst the other was discarded because of extensive endothelial cell necrosis noted after 15 days in organ culture. CONCLUSION: HSV within a donor cornea may cause endothelial destruction in organ culture and both primary graft failure and ulcerative keratitis after transplantation. Endothelial necrosis of a donor cornea in culture also raises the possibility of HSV infection within the fellow cornea.
Assuntos
Sobrevivência de Enxerto , Herpes Simples/transmissão , Ceratoplastia Penetrante/métodos , Simplexvirus/isolamento & purificação , Adulto , Idoso , Idoso de 80 Anos ou mais , Endotélio Corneano/patologia , Feminino , Distrofia Endotelial de Fuchs/cirurgia , Distrofia Endotelial de Fuchs/virologia , Humanos , Ceratocone/cirurgia , Ceratocone/virologia , Masculino , Necrose , Reação em Cadeia da Polimerase , Simplexvirus/genética , Simplexvirus/imunologiaRESUMO
PURPOSE: To develop a multiplex polymerase chain reaction (PCR) for the detection of adenovirus, herpes simplex virus, and Chlamydia trachomatis in conjunctival swabs. METHODS: Oligonucleotide primers for detection of the 3 agents were combined in one reaction and evaluated for optimal performance using control DNAs of adenovirus type 2, herpes simplex virus, and C. trachomatis plasmid. The multiplex PCR was evaluated prospectively against its corresponding uniplex PCRs, virus isolation, Chlamydia Amplicor PCR, and an immunoassay technique (immune dot blot test) in a total of 805 conjunctival swabs from patients with suspected viral and chlamydial keratoconjunctivitis. RESULTS: The multiplex PCR was as sensitive as uniplex PCRs for the detection of the agents in clinical specimens. In the prospective study, 48 of 49 (98%) clinical specimens were positive for adenovirus by the multiplex PCR compared with 26 of 49 (53%) by adenovirus isolation. For herpes simplex virus detection, the multiplex PCR had a sensitivity of 92% (34/37) compared with 94.5% (35/37) by cell culture. The multiplex PCR produced identical results to the Amplicor PCR (21/21; 100%) compared with 71% (15/21) by the immune dot blot test. CONCLUSIONS: With clinical specimens the multiplex PCR was as sensitive as its respective uniplex PCRs but more sensitive than adenovirus isolation and as sensitive as herpes simplex virus isolation or C. trachomatis Amplicor PCR. It has the potential to replace several diagnostic tests with consequent savings in cost. The test also reduces the risk of misdiagnosis by the clinicians.
Assuntos
Infecções por Adenovirus Humanos/diagnóstico , Infecções por Chlamydia/diagnóstico , Infecções Oculares/diagnóstico , Infecções por Herpesviridae/diagnóstico , Ceratoconjuntivite/diagnóstico , Reação em Cadeia da Polimerase/métodos , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/genética , Adenovírus Humanos/isolamento & purificação , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/genética , Chlamydia trachomatis/isolamento & purificação , Túnica Conjuntiva/virologia , Primers do DNA/química , DNA Viral/análise , Infecções Oculares/virologia , Infecções por Herpesviridae/virologia , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/isolamento & purificação , Humanos , Ceratoconjuntivite/microbiologia , Estudos Prospectivos , Sensibilidade e EspecificidadeRESUMO
PURPOSE: In a previous investigation it was demonstrated that circulating antibodies to a 66-kDa corneal epithelial antigen (BCEA-A) are associated with peripheral ulcerative keratitis (PUK) in patients with Wegener's granulomatosis (WG). The aim of this study was to identify BCEA-A. METHODS: The 66-kDa antigen was purified from a bovine corneal epithelial protein extract, using DE52 ion exchange chromatography. Purified protein was used to raise rabbit polyclonal antibodies. These antibodies were used to screen a bovine corneal epithelial cDNA expression library. Positive clones were purified and sequenced. Clones were identified by DNA sequence homology searches of the GenBank DNA database. RESULTS: A cDNA clone that demonstrated strong binding to both the rabbit polyclonal antibody and patient sera, showed 85% homology to rabbit cytokeratin 3 (K3). K3 is a basic cytokeratin specific to corneal epithelium. No bovine DNA sequence for K3 is available. However, bovine K3 is larger than rabbit K3, with a molecular weight of 66 kDa. Immunofluorescence using both patient sera and the rabbit antibody demonstrated a cytoplasmic binding pattern on human corneal epithelium. CONCLUSIONS: This evidence suggests that the 66-kDa autoantigen (BCEA-A) associated with PUK in WG is cytokeratin 3, and this may form the basis of a diagnostic/prognostic test.
Assuntos
Autoantígenos/metabolismo , Úlcera da Córnea/metabolismo , Epitélio Corneano/metabolismo , Granulomatose com Poliangiite/metabolismo , Queratinas/metabolismo , Sequência de Aminoácidos , Animais , Bovinos , Cromatografia por Troca Iônica , Primers do DNA/química , Eletroforese em Gel de Poliacrilamida , Epitélio Corneano/química , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Queratinas/genética , Queratinas/isolamento & purificação , Dados de Sequência Molecular , Peso Molecular , Coelhos , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
PURPOSE: To evaluate newly designed primers in a polymerase chain reaction (PCR) for the detection of adenovirus DNA in conjunctival swabs. METHODS: Oligonucleotides were derived from the adenovirus hexon gene and modified such that a maximum of only two mismatches occurred with adenovirus types 2 through 5, 7, and 16. Specificity was determined against adenovirus types 2 through 4, 7, 8 through 11, 14, 19, 37, 40, and 41 and from non-adenoviral DNA and the sensitivity by PCR amplification of purified adenovirus type 2 DNA. The assay was compared retrospectively with cell culture and a PCR with different primers on 59 stored conjunctival swab samples. The new PCR also was used prospectively in comparison with cell culture on 2743 conjunctival swabs. RESULTS: The 140-bp product was amplified from all the adenovirus serotypes tested except types 40 and 41, which have not been isolated from the eye. There were no amplified products from the non-adenoviral DNA tested. With adenovirus type 2 DNA, despite two deliberate mismatches, 40 copies of the target were detectable after PCR and ethidium bromide-staining. In the retrospective study, 51 of 55 (92.7%) were positive by this new PCR compared with 42 of 55 (76.4%) by the older PCR and 40 of 55 (72.7%) by cell culture. In the prospective study, the new PCR detected 386 of 415 (93%) adenovirus-positive specimens compared with 248 of 415 (59.8%) by cell culture. Of 167 specimens positive for herpes simplex virus by cell culture, none were positive by the adenovirus PCR. CONCLUSIONS: PCR with the newly designed primers shows a much increased sensitivity over cell culture and previous PCRs for the detection of adenoviruses in conjunctival swabs.
Assuntos
Infecções por Adenovirus Humanos/diagnóstico , Adenovírus Humanos/genética , Conjuntivite Viral/diagnóstico , DNA Viral/análise , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/isolamento & purificação , Células Cultivadas , Túnica Conjuntiva/citologia , Túnica Conjuntiva/virologia , Conjuntivite Viral/virologia , Primers do DNA/química , Humanos , Reação em Cadeia da Polimerase/métodos , Estudos Prospectivos , Estudos Retrospectivos , Sensibilidade e EspecificidadeRESUMO
PURPOSE: In a previous investigation into corneal autoimmunity, it was demonstrated that a putative autoantigen, a protein of 66 kDa, present in bovine corneal epithelium, binds circulating autoantibodies in approximately 60% of patients with Wegener's granulomatosis (WG). The aim of the present study was to characterize and identify the 66-kDa protein. METHODS: A purification protocol was established for the 66-kDa protein using standard chromatography techniques. During the purification procedure it became clear that the 66-kDa protein detected in patients' sera was in fact two proteins, both running at 66 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, that eluted in different fractions on DE-52 chromatography columns. These two proteins have been labeled bovine corneal epithelial antigen-A and -B (BCEA-A and BCEA-B). Further investigations of antibody binding have demonstrated that patients' sera bind to either one or the other of these proteins with no cross-reactivity between them. Separated BCEA-A and BCEA-B protein extracts were immunoblotted with 27 WG patients' sera, 10 Churg-Strauss syndrome (CSS) patients' sera, 31 rheumatoid arthritis (RA) patients' sera, and 40 healthy control subjects' sera from the blood bank. RESULTS: Forty-six percent of WG patients' sera had antibodies to one of the 66-kDa antigens, whereas none of the healthy control subjects' sera had 66-kDa antibodies (P < 10(-5)). In the WG group, 31% were positive to BCEA-A (versus controls, P = 0.0023), and 15% were positive to BCEA-B. WG patients with peripheral ulcerative keratitis (PUK) had a significant association with anti-BCEA-A antibodies when compared with healthy control subjects (50%, P < 10(-6)). However, in the RA group with no eye disease there was an association with BCEA-A (25%, P = 0.011) but not in the RA group with PUK. The frequency of anti-BCEA-B antibodies was significantly increased in patients with CSS (60%, P < 10(-7)). CONCLUSIONS: In summary, it has been shown that vasculitis patients have antibodies to two 66-kDa corneal antigens and that autoantibodies to these antigens are mutually exclusive. It has also been shown that antibodies to BCEA-B are associated with CSS, whereas BCEA-A antibodies are associated with WG and RA.
Assuntos
Autoanticorpos/análise , Autoantígenos/metabolismo , Conjuntivite/metabolismo , Úlcera da Córnea/metabolismo , Epitélio Corneano/metabolismo , Vasculite/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Artrite Reumatoide/imunologia , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Autoantígenos/imunologia , Autoantígenos/isolamento & purificação , Bovinos , Conjuntivite/imunologia , Conjuntivite/patologia , Úlcera da Córnea/imunologia , Úlcera da Córnea/patologia , Reações Cruzadas , Eletroforese em Gel Bidimensional , Epitélio Corneano/química , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Immunoblotting , Pessoa de Meia-Idade , Peso Molecular , Coelhos , Vasculite/imunologia , Vasculite/patologiaRESUMO
AIMS/BACKGROUND: In adult tissues the expression of tenascin-cytotactin (TN-C), an extracellular matrix glycoprotein, is limited to tumours and regions of continuous renewal. It is also transiently expressed in cutaneous and corneal wound healing. There are limited data regarding its expression in inflammation and scarring of the adult human cornea. In this study, TN-C expression patterns in normal, inflamed, and scarred human corneas have been examined. METHODS: Penetrating keratoplasty specimens were selected from cases of herpes simplex keratitis, herpes zoster ophthalmicus, rheumatoid arthritis ulceration, bacterial keratitis, rosacea keratitis, interstitial keratitis, and previous surgery so as to encompass varying degrees of active and chronic inflammation and scarring. TN-C in these and in normal corneas was immunodetected using TN2, a monoclonal antibody to human TN-C. RESULTS: There was no TN2 immunopositivity in normal corneas except at the corneoscleral interface. In pathological corneas, TN2 immunopositivity was localised in and around regions of active inflammation, fibrosis, and neovascularisation. TN2 positivity was less in acute inflammation than in active chronic inflammation. Mature, avascular scar tissue and epithelial downgrowth were TN2 negative. CONCLUSION: These results indicate that in the adult human cornea, TN-C expression is induced in regions of inflammation, fibrosis, and neovascularisation, but that expression is absent in mature, avascular scar tissue. This suggests a role for this glycoprotein in inflammation, healing, and extracellular matrix reorganisation of the cornea.
Assuntos
Cicatriz/metabolismo , Doenças da Córnea/metabolismo , Tenascina/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Criança , Pré-Escolar , Cicatriz/patologia , Córnea/metabolismo , Doenças da Córnea/patologia , Humanos , Imuno-Histoquímica , Ceratite/metabolismo , Pessoa de Meia-IdadeRESUMO
Adenoviruses of subgenus F (types 40 and 41) cause infantile gastroenteritis and adenoviruses principally of types 1-7 are found in feces during respiratory or generalized infections. Adenoviruses (mostly types 3, 4, 8, 19, or 37) are also linked with follicular or epidemic conjunctivitis. The diagnostic efficiency of the polymerase chain reaction (PCR) for adenoviruses was assessed using genus-reactive primers H1 and H2 or JCH1 and JCH2 or subgenus F-specific primers F1a and F2a. With diarrheal stool specimens containing subgenus F adenoviruses, F1a/F2a PCR achieved at least as high a positivity rate (75/76 [99%]) as electron microscopy (72/76 [95%]) and was more sensitive than polyclonal antibody-based immune electron microscopy (IEM) for subgenus identification (75/76 [99%] vs. 66/76 [87%], P = 0.008). Twenty-three subgenus F strains untypeable by monoclonal antibody-based IEM were typed as 40 (n = 4) or 41 (n = 19) by Hha I digestion of the PCR product. The genus-reactive primer pairs provided DNA amplification assays of generally equal efficiency on conjunctival swab specimens though possible nucleic acid degradation in DNA extracts during storage could have meant that JCH1/JCH2 PCR was truly the more sensitive. The use of either genus-reactive primer set on fecal specimens cannot be recommended because, although the positivity rates with subgenus F PCR positive specimens were high (70/75 [93%] for H1 and H2, 14/15 [93%] for JCH1 and JCH2), the detection rates were disappointing with similar specimens yielding nonsubgenus F adenoviruses.
Assuntos
Infecções por Adenoviridae/virologia , Adenovírus Humanos/genética , Oftalmopatias/virologia , Reação em Cadeia da Polimerase/métodos , Adenovírus Humanos/classificação , Adenovírus Humanos/isolamento & purificação , Animais , Chlorocebus aethiops , Primers do DNA , Olho/virologia , Fezes/virologia , Humanos , Estudos Prospectivos , Estudos Retrospectivos , Sensibilidade e Especificidade , Células Tumorais Cultivadas , Células VeroRESUMO
Adenoviruses and herpes simplex virus (HSV) can cause clinically indistinguishable episodes of acute eye disease. Adenovirus infection is associated with nosocomial outbreaks and HSV may result in episodes of recurrent ocular inflammation. In a comparison of multiplex PCR for the two viral DNAs and virus isolation in cell culture, identical results were obtained for 18 of 20 specimens (positive for adenovirus in 5, HSV in 5, and negative in 8). One specimen was falsely negative for each viral DNA. Inclusion of human beta-globin primers in the adenovirus-HSV reaction was precluded by a consequential 10--100-fold reduction in sensitivity for the two viral targets and by the failure of beta-globin DNA amplification at the annealing temperature (45 degrees C) required to ensure detection of adenoviruses of serotypes 7 and 11 with the selected adenovirus primers. A single-target beta-globin PCR gave positive results with 19 of the 20 specimens prepared by treatment with proteinase K lysis buffer, indicating the effectiveness of this simple DNA extraction procedure. Nonetheless, the availability of effective antiviral therapy for HSV made monitoring for extraction failure using human primers crucial to avoid false-negative results for HSV DNA. Adenovirus-HSV PCR has considerable potential for the rapid diagnosis of viral eye disease particularly if beta-globin primers can be included in the reaction.
Assuntos
Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/isolamento & purificação , Infecções Oculares Virais/virologia , Olho/virologia , Reação em Cadeia da Polimerase/métodos , Simplexvirus/isolamento & purificação , Infecções por Adenovirus Humanos/diagnóstico , Sequência de Bases , DNA Viral/análise , Olho/patologia , Infecções Oculares Virais/diagnóstico , Humanos , Ceratite Herpética/diagnóstico , Ceratite Herpética/virologia , Dados de Sequência MolecularRESUMO
Adenoviruses are associated with endemic and epidemic acute conjunctivitis, large nosocomial outbreaks reflecting virus transmission on unwashed hands or inadequately sterilised ophthalmic instruments. The polymerase chain reaction (PCR) proved more sensitive than antigen detection by immune dot-blot test for the rapid diagnosis of ocular adenovirus infection (sensitivities in a retrospective study 112/123 (91%) versus 72/123 (59%), P < 0.001). Indeed, in a prospective comparison, DNA amplification and virus isolation generated similar numbers of positive results (34 versus 32), though five PCR positive results were possibly false positives. The sensitivity of the PCR was largely independent of adenovirus subgenus or serotype, though reduced sensitivity with subgenus B strains could not be excluded. Specimen preparation for DNA amplification using a simple lysis buffer proved more effective than phenol-chloroform extraction. The immune dot-blot test gave unavoidable false positive results, but with the PCR this problem could be minimized by technical modifications. The PCR could replace antigen detection and virus isolation as the initial test for adenoviruses in conjunctival swabs, with cell culture only being retained for adenovirus serotyping in PCR positive specimens and for other viruses such as herpes simplex.