RESUMEN
BACKGROUND: T cells that express Cutaneous Lymphocyte-Associated antigen (CLA) have the potential of migrating to the skin, and are hypothesized to play a role in cutaneous atopic disease. AIM: To investigate the immune phenotype and cytokine responses to Der p 1 stimulation of CLA+ T cells in extrinsic atopic dermatitis (EAD). DESIGN: In vitro testing, with controls. METHODS: Peripheral blood mononuclear cells (PBMC) were obtained from EAD patients (n=27) and non-atopic healthy individuals (n=22). Phenotypic analysis of naive, CLA+ and non-CLA+ memory/effector CD4+ and CD8+ T cells used markers of cell activation, differentiation, adhesion, apoptosis and chemokine receptor expression. Cytokine responses in these cells were studied following Der p 1 stimulation. RESULTS: CLA+ T cells from EAD patients expressed significantly higher levels of CD25, HLA-DR, CD38, CD71, CXCR1, CXCR2 and lower levels of bcl2, CCR5, CCR7, CXCR3, and CD62L (p<0.05). DISCUSSION: In EAD patients, CLA+ T cells express increased levels of markers associated with activation, adhesion and apoptosis, show differences in the level of expression of differentiation markers and display a distinct chemokine receptor preference, compared with cells from healthy controls. These data suggest a significant role for CLA+ T cells in the pathogenesis of cutaneous atopic disease.
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Antígenos de Neoplasias/inmunología , Dermatitis Atópica/inmunología , Glicoproteínas de Membrana/inmunología , Receptores Mensajeros de Linfocitos/análisis , Piel/inmunología , Linfocitos T/inmunología , Adulto , Antígenos Dermatofagoides/inmunología , Antígenos de Diferenciación de Linfocitos T , Proteínas de Artrópodos , Cisteína Endopeptidasas , Femenino , Humanos , MasculinoRESUMEN
BACKGROUND: Several studies have investigated levels of T-cell-derived interleukin (IL)-10 in individuals with atopic dermatitis, with conflicting results. AIMS/HYPOTHESIS: In order to address whether stratification of disease severity may help resolve the different findings, the hypothesis was tested that individuals with severe atopic dermatitis have a lower frequency of circulating IL-10-producing, allergen-specific CD4+ T cells than do individuals with mild disease. METHODS: Using peripheral blood mononuclear cells derived from individuals with severe (n=12) and mild atopic dermatitis (n=10) and from nonatopic controls (n=10), we investigated production by CD4+ T cells of tumour necrosis factor (TNF)-alpha, IL-4, IL-5, IL-13 and IL-10 in response to phorbol myristate acetate/ionomycin and Der p1 allergen. RESULTS: It was observed that there were significantly higher frequencies of allergen-specific circulating CD4+ T cells producing TNF-alpha- IL-4-, IL-5- and IL-13, and lower frequencies of these cells producing IL-10 in individuals with severe atopic dermatitis compared with mildly affected individuals and nonatopic controls (P<0.01 for all comparisons). Furthermore, the Der p1-specific CD4+ T cells were enriched within the subset of cells positive for cutaneous lymphocyte-associated antigen. CONCLUSIONS: Analysis of levels of allergen-specific CD4+ T-cell production of IL-10 in relation to disease severity argues in favour of a role for IL-10 in the control of atopic dermatitis.
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Linfocitos T CD4-Positivos/citología , Dermatitis Atópica/inmunología , Interleucina-10/biosíntesis , Linfocitos T CD4-Positivos/inmunología , Citometría de Flujo/métodos , Humanos , Interleucinas/biosíntesis , Estimulación QuímicaRESUMEN
Factors controlling the expression of cutaneous lymphocyte-associated antigen (CLA) by T cells are poorly understood, but data from murine and human CD4(+) T cell systems have suggested that cytokines play an important role. However, there are no data examining the influence of cytokines on the expression of CLA by human antigen-specific CD8(+) T cells. Peripheral blood mononuclear cells (PBMC) were isolated from 10 HLA-A*0201-positive healthy individuals. Using HLA-peptide tetrameric complexes refolded with immunodominant peptides from Epstein-Barr virus (EBV), cytomegalovirus (CMV) and influenza A virus, we investigated the temporal associations of CLA expression by viral-specific CD8(+) T cells following stimulation with antigen. Ex vivo influenza matrix-specific CD8(+) T cells expressed significantly (P < 0.05) greater levels of CLA than EBV BMLF1 and CMV pp65-specific CD8(+) T cells (mean 9.7% influenza matrix versus 1.4% BMLF1 versus 1.1% pp65) and these differences were sustained on culture. However, regardless of viral specificity, interleukin (IL)-12 and IL-4 induced significant (P < 0.05) dose-dependent up-regulation and down-regulation of CLA expression, respectively, with IL-4 showing a dominant negative effect. In many cases, IL-4 resulted in complete abrogation of detectable CLA expression by the viral-specific CD8(+) T cells. Overall these data demonstrate that CLA expression by human viral-specific CD8(+) T cells is highly dynamic and that IL-4 causes significant down-regulation. Disorders associated with a type 2 cytokine shift may reduce the efficiency of skin homing by viral-specific CD8(+) T cells. Furthermore, the ability to modify the local and systemic microenvironment may offer novel therapeutic strategies that influence tissue-specific T cell homing.
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Linfocitos T CD8-positivos/inmunología , Interleucina-4/inmunología , Glicoproteínas de Membrana/metabolismo , Receptores Mensajeros de Linfocitos/metabolismo , Enfermedades Cutáneas Virales/inmunología , Antígenos de Diferenciación de Linfocitos T , Antígenos de Neoplasias , Células Cultivadas , Citomegalovirus/inmunología , Dermatitis Atópica/complicaciones , Dermatitis Atópica/inmunología , Regulación hacia Abajo/inmunología , Epítopos de Linfocito T/inmunología , Citometría de Flujo/métodos , Herpesvirus Humano 4/inmunología , Humanos , Interleucina-12/inmunología , Orthomyxoviridae/inmunología , Piel/inmunología , Enfermedades Cutáneas Virales/etiologíaRESUMEN
STUDY DESIGN: A basic science animal investigation. OBJECTIVES: To determine if bipedal rats differ in upright posture compared with quadrupedal rats. SUMMARY OF BACKGROUND DATA: It has been reported that surgically induced bipedalism in the rat leads to habitual upright posture. Based on this finding, bipedal rats have been used to study the changes erect posture induces in bone, ligament, muscle, and intervertebral discs. Previous studies have used direct observation as a means to describe posture. This study is the first to quantify postural differences between bipedal and quadrupedal rats. METHODS: Eleven bipedal rats were created by forelimb and tail amputation within 24 hours of birth. Eleven quadrupedal rats served as controls. Specialized cages were used with infrared sensors, and a computer program measured the total amount of time in the upright stance, the number of stands, and the amount of horizontal movement in the upright stance. Statistical comparisons were made between bipedal and quadrupedal rats hourly and over a 24-hour period of time. RESULTS: Quadrupedal rats assumed an upright posture for a significantly greater amount of time than bipedal rats when monitored over 24 hours (P = 0.016). Quadrupedal and bipedal rats did not differ in the number of stands (P = 0.63) or in the amount of horizontal movement in the upright stance (P = 0.34) over 24 hours. Similar results were obtained when comparing hourly intervals. CONCLUSION: This study quantifiably indicates that bipedal rats do not assume a more erect posture and spend no more time in an upright position compared with quadrupedal rats. The upright posture may not be the cause of some previously reported anatomic changes observed in the bipedal rat.
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Movimiento/fisiología , Orientación/fisiología , Postura/fisiología , Amputación Quirúrgica , Animales , Conducta Animal , Femenino , Miembro Anterior/cirugía , Embarazo , Ratas , Ratas WistarAsunto(s)
Electrocirugia/métodos , Papillomaviridae/genética , Infecciones por Papillomavirus/cirugía , Infecciones Tumorales por Virus/cirugía , Displasia del Cuello del Útero/virología , Neoplasias del Cuello Uterino/virología , Adolescente , Adulto , Colposcopía , Conización , ADN Viral/análisis , Femenino , Humanos , Persona de Mediana Edad , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/cirugía , Displasia del Cuello del Útero/patología , Displasia del Cuello del Útero/cirugíaRESUMEN
HIV-positive patients are at increased risk of developing adenovirus infection, particularly of the gastrointestinal tract and with unusual subgenus D strains. To investigate humoral immunity to these strains of adenoviruses, the humoral immune response was examined in longitudinal samples of serum against isolates collected from a prospective study of HIV-positive patients with subgenus D adenovirus infection. Of 10 HIV-positive patients developing adenovirus infection, 3 had chronic infection (8->27 months) with one serotype, 3 had chronic infection (>/=10 months) with changing serotypes and 4 had acute and self-limiting adenovirus infection (<1 month). Fifty-one sera were tested, and samples collected before adenovirus infection were available in 8 patients. Neutralising assays were performed against the patient's own isolate (adenoviruses 9, 17, 19, 19/23, 19/37, 23, 26, 23/26, 43 and 46) and common circulating strains of adenovirus 1-5. Indirect immunofluorescence tests were carried out against the autologous isolate and complement-fixation tests were undertaken using a standard assay. Immunofluorescence test antibodies were detected (titre >/=160) in all patients, and present to high titre (>/=320) in 8/10 patients. Complement-fixing antibodies were not detected in significant titre. Of particular note, there was no significant neutralising antibody response to the patient's own isolate after acute infection. Neutralising antibody to adenovirus 3 (titre 20) was transiently detected in two patients. In the remaining patients neutralising antibody directed against adenoviruses 1-5 was not detected. Persistent carriage of subgenus D adenoviruses in HIV-positive patients is probably the result of failure of cell-mediated immune responses to clear primary infection. Nevertheless, there is marked impairment of B cell responses resulting in poor neutralising and complement-fixing antibody production even though immunofluorescence test determined antibodies are produced in high titre. These possibly reflect impairment of effective B cell priming mechanisms within the germinal centres of lymph nodes, or the polyclonal activation of B cells driven by HIV infection.
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Infecciones Oportunistas Relacionadas con el SIDA/inmunología , Infecciones por Adenovirus Humanos/inmunología , Anticuerpos Antivirales/inmunología , Infecciones Oportunistas Relacionadas con el SIDA/sangre , Infecciones Oportunistas Relacionadas con el SIDA/virología , Infecciones por Adenovirus Humanos/sangre , Infecciones por Adenovirus Humanos/complicaciones , Infecciones por Adenovirus Humanos/virología , Adenovirus Humanos/inmunología , Adenovirus Humanos/aislamiento & purificación , Anticuerpos Antivirales/sangre , Pruebas de Fijación del Complemento , Técnica del Anticuerpo Fluorescente Indirecta , Humanos , Pruebas de Neutralización , Estudios ProspectivosRESUMEN
Subgenus identification of adenoviruses is of clinical importance and is as informative as identification by serotype in most clinical situations. A PCR-based identification of adenovirus subgenera A, B, C, D, E, and F and sometimes serotypes is described. The PCR uses nonnested primer pair ADRJC1-ADRJC2, which targets a highly conserved region of the adenovirus hexon gene, has a sensitivity of 10 to 40 copies of adenovirus type 2 (Ad2) DNA, and generates 140-bp PCR products from adenovirus serotypes representative of all the subgroups. The PCR products of all subgroups can be differentiated on the basis of the restriction fragment patterns produced by a total of five restriction endonucleases. In addition, serotypes Ad40 and Ad41 (subgroup F) and important serotypes of subgroup D (Ad8, Ad10, Ad19, and Ad37) can easily be differentiated, but serotypes within subgroups B and C cannot. The method was assessed by blind subgenus identification of 56 miscellaneous clinical isolates of adenoviruses. The identities of these isolates at the subgenus level by the PCR correlated 91% (51 of 56) with the results of serotyping by the neutralization test, and 9% (5 of 56) of clinical isolates produced discordant results.
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Adenovirus Humanos/clasificación , Proteínas de la Cápside , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Infecciones por Adenovirus Humanos/virología , Adenovirus Humanos/genética , Secuencia de Bases , Cápside/genética , Secuencia Conservada , Cartilla de ADN , ADN Viral/aislamiento & purificación , Humanos , Datos de Secuencia Molecular , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido NucleicoRESUMEN
OBJECTIVES: To determine the prevalence of high grade anal intraepithelial neoplasia (HGAIN), the value of anal cytology in screening for HGAIN, and the characterisation of epidemiological factors and human papillomavirus (HPV) types. METHODS: Prospective cohort study of HIV positive homosexual men. Subjects were interviewed, underwent STD, anal cytological, and HPV screening at enrolment and at subsequent follow up visits with anoscopy and biopsy at the final visit. 57 enrolled, average CD4 count 273 x 10(6)/l (10-588); 41 completed the cytological surveillance over the follow up period (181 visits, average follow up 17 months), 38 of these had anoscopy and anal biopsy. RESULTS: Oncogenic HPV types were detected in 84% and high grade dyskaryosis in 10.5% (6/57) at enrollment. There was a 70% incidence of high grade dyskaryosis during follow up in patients with negative/warty or low grade dyskaryosis at enrollment. Anoscopy correlated with histology in high grade AIN lesions (sensitivity 91%, specificity 54%) and cytology was 78% sensitive (18/23) for HGAIN on biopsy. CONCLUSIONS: AIN and infection with multiple oncogenic HPV types are very common among immunosuppressed HIV positive homosexual men. Apparent progression from low to high grade cytological changes occurred over a short follow up period, with no cases of carcinoma. All 23 cases of HGAIN were predicted by cytology and/or anoscopy. Future studies focusing on the risk of progression to carcinoma are needed before applying anal cytology as a screening tool for AIN in this population.
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Neoplasias del Ano/virología , Carcinoma in Situ/virología , Infecciones por VIH/complicaciones , Homosexualidad Masculina , Papillomaviridae , Infecciones por Papillomavirus/complicaciones , Adulto , Neoplasias del Ano/epidemiología , Neoplasias del Ano/patología , Carcinoma in Situ/epidemiología , Carcinoma in Situ/patología , Estudios de Cohortes , ADN Viral/análisis , Infecciones por VIH/epidemiología , Homosexualidad Masculina/estadística & datos numéricos , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Papillomaviridae/genética , Infecciones por Papillomavirus/epidemiología , Infecciones por Papillomavirus/virologíaRESUMEN
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.
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Infecciones por Adenovirus Humanos/diagnóstico , Adenovirus Humanos/genética , Conjuntivitis Viral/diagnóstico , ADN Viral/análisis , Infecciones por Adenovirus Humanos/virología , Adenovirus Humanos/aislamiento & purificación , Células Cultivadas , Conjuntiva/citología , Conjuntiva/virología , Conjuntivitis Viral/virología , Cartilla de ADN/química , Humanos , Reacción en Cadena de la Polimerasa/métodos , Estudios Prospectivos , Estudios Retrospectivos , Sensibilidad y EspecificidadRESUMEN
We evaluated Cobas Amplicor, a highly automated polymerase chain reaction (PCR) system, to test first-void urine (FVU) and urethral swab specimens for Chlamydia trachomatis and Neisseria gonorrhoeae in men attending a sexually transmitted infection (STI) clinic. Results were compared against an in-house radioimmune dot blot (DB) test for C. trachomatis and selective culture for N. gonorrhoeae. Three hundred and ninety sets of specimens were obtained from 378 consecutive new and returned-new patients. Gonorrhoea prevalence was 9.49%, with no significant difference in sensitivity or specificity between culture and PCR. Chlamydia prevalence was 15.4%, with sensitivities of: DB 55%, PCR of FVU 86.7%, urethral swab PCR 90%. The specificity of PCR on FVU and urethral swabs was 100%. We have shown that Cobas Amplicor PCR is highly sensitive and specific in the diagnosis of chlamydia and gonorrhoea in men attending an STI clinic. Further economic and scientific studies are needed to determine the cost-effectiveness of this technique for screening in primary care settings.
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Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/aislamiento & purificación , Enfermedades de los Genitales Masculinos/diagnóstico , Gonorrea/microbiología , Neisseria gonorrhoeae/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Automatización , Infecciones por Chlamydia/patología , Infecciones por Chlamydia/orina , Chlamydia trachomatis/genética , Enfermedades de los Genitales Masculinos/microbiología , Enfermedades de los Genitales Masculinos/patología , Enfermedades de los Genitales Masculinos/orina , Gonorrea/patología , Gonorrea/orina , Humanos , Masculino , Neisseria gonorrhoeae/genéticaRESUMEN
Four fluorescent antibody reagents were evaluated for their suitability for the identification of adenovirus isolates by immunofluorescence. The antibodies used in the reagents consist of monoclonal antibodies against adenovirus type 3 (Ad3), Ad4, Ad8, and adenoviruses of subgroup C (Ad1,2,5,6), serotypes known to occur in outbreaks of disease. Most of the monoclonal antibodies employed were reactive against type-specific antigens found on the hexon protein. Reagents employing two noncompeting anti-hexon antibodies were more sensitive than reagents prepared with only one monoclonal antibody, although both types of reagents exhibited a high degree of specificity. Five hundred and seventeen adenovirus isolates (359 of which had previously been typed by other methods) and 46 nonadenovirus isolates were examined with all four type-specific reagents in parallel with an adenovirus group-specific reagent. The results indicate that direct typing of adenovirus isolates is feasible, leading to significant savings in time compared to other typing methods and should contribute to the management of certain adenovirus infections, particularly during outbreaks.
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Infecciones por Adenoviridae/diagnóstico , Adenovirus Humanos/clasificación , Adenovirus Humanos/aislamiento & purificación , Proteínas de la Cápside , Infecciones por Adenoviridae/epidemiología , Anticuerpos Monoclonales , Antígenos Virales/inmunología , Cápside/inmunología , Diagnóstico Diferencial , Brotes de Enfermedades , Técnica del Anticuerpo Fluorescente Directa , Humanos , Manejo de EspecímenesRESUMEN
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.
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Infecciones por Adenoviridae/virología , Adenovirus Humanos/genética , Oftalmopatías/virología , Reacción en Cadena de la Polimerasa/métodos , Adenovirus Humanos/clasificación , Adenovirus Humanos/aislamiento & purificación , Animales , Chlorocebus aethiops , Cartilla de ADN , Ojo/virología , Heces/virología , Humanos , Estudios Prospectivos , Estudios Retrospectivos , Sensibilidad y Especificidad , Células Tumorales Cultivadas , Células VeroRESUMEN
Thirty-three cadaveric dissections were performed to identify radial nerve branching patterns to the triceps brachii. Radial innervation of the long head of the triceps originated in the axilla in 88% of the cases and the brachio-axillary angle in 12%. Innervation of the medial head of the triceps originated in the spiral groove in 52% of the cases, the brachio-axillary angle in 39%, and the axilla in 9%. The lateral head was innervated by branches arising in the spiral groove in 70% of the cases, the brachio-axillary angle in 24%, and the axilla in 6%. On average, the radial nerve crossed the midline in the proximal 45% of the arm, 3 cm superior to the level of the deltoid insertion. An intramuscular tendon was present in the medial head of the triceps. The tendon, located medial to the midline of the arm, was seen in all specimens. This tendon serves as an interneural plane with nerve branches descending on either side, but never crossing from one side to the other. Due to the complexity of radial nerve branching, this tendon may be used as a reference plane for longitudinal splitting of the medial head minimizing the risk of nerve damage.
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Brazo/inervación , Nervio Radial/anatomía & histología , Humanos , Músculos/inervación , Tendones/anatomía & histologíaRESUMEN
Endemic (type 1, 2, 5 and 6) and epidemic (type 3, 4 and 7) respiratory adenovirus infections are associated with upper respiratory tract symptoms, pharyngoconjunctival fever, and pneumonia. Improved methods of diagnosis are needed, particularly in immunocompromized patients. We examined 93 throat swabs or nasopharyngeal aspirates from patients with acute respiratory disease using virus isolation and an adenovirus-specific polymerase chain reaction (PCR) based on consensus primers H1 and H2 derived from the hexon region DNA sequences of serotypes 2 and 5. Specimens which yielded viruses other than adenovirus in cell culture (n = 23) or which were negative for infectious viruses (n = 25) were negative in the PCR. The sensitivity of DNA amplification was 76% (34/45) in comparison with virus culture, being markedly lower with subgenus B (types 3 and 7) strains than with subgenus C (type 1, 2, 5 and 6) isolates (8/16 (50%)) vs. 26/28 (93%). P = 0.004) despite the use of a low annealing temperature to maximize detection of adenoviruses belonging to subgenera other than C. Of the 11 samples falsely negative in a single-round PCR but yielding adenovirus type 1 (n = 1), type 2 (n = 1). type 3 (n = 7), type 7 (n = 1), or untyped isolates (n = 1) in cell culture, nine (82%) gave positive results after nested DNA amplification. Possible approaches to further improving the performance of adenovirus PCR with respiratory specimens are discussed.
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Infecciones por Adenoviridae/diagnóstico , ADN Viral/análisis , Reacción en Cadena de la Polimerasa , Infecciones del Sistema Respiratorio/diagnóstico , Reacciones Falso Positivas , Humanos , Sensibilidad y EspecificidadRESUMEN
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.
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Infecciones por Adenovirus Humanos/virología , Adenovirus Humanos/aislamiento & purificación , Infecciones Virales del Ojo/virología , Ojo/virología , Reacción en Cadena de la Polimerasa/métodos , Simplexvirus/aislamiento & purificación , Infecciones por Adenovirus Humanos/diagnóstico , Secuencia de Bases , ADN Viral/análisis , Ojo/patología , Infecciones Virales del Ojo/diagnóstico , Humanos , Queratitis Herpética/diagnóstico , Queratitis Herpética/virología , Datos de Secuencia MolecularRESUMEN
Right and left upper pulmonary venous flow is usually assessed with monoplane transesophageal echocardiography (TEE) in the transverse imaging plane. Pulmonary venous flow in the transverse imaging plane may be relatively difficult to record because of the larger angle between the pulmonary vein and the transducer beam. To compare the quality of echocardiographically derived Doppler flows of the right and left upper pulmonary veins between the longitudinal and transverse imaging planes with TEE, we performed pulsed-wave Doppler TEE of both upper pulmonary veins in transverse and longitudinal imaging planes in 36 patients with various diseases. We also recorded a quality index for each flow profile and the angle between the transducer beam and the pulmonary vein. The quality index of the left pulmonary venous flow assessed with the longitudinal and transverse imaging planes was similar in 35 (95%) of 36 patients, whereas the longitudinal imaging plane was superior to the transverse plane in one patient (3%). In contrast, the quality index of the right pulmonary venous flow assessed with the longitudinal and transverse imaging planes was similar in only 19 (53%) of 36 patients, whereas in 17 patients (47%) the longitudinal imaging plane was superior to the transverse imaging plane. The quality index had a significant effect on the Doppler flow recordings; suboptimal-quality flow recordings significantly underestimated the pulmonary venous diastolic flow integrals. The left atrium was larger in those patients with unobtainable flows than in those patients with exclusively obtainable flows (p < 0.001). The angle between the sample volume and the right pulmonary vein was larger in the transverse imaging plane than in the longitudinal plane (p < 0.001). In conclusion, the longitudinal imaging plane is generally superior to the transverse imaging plane for assessing right pulmonary venous flow and is recommended for performing a comprehensive assessment of pulmonary venous flow. The ability to obtain quality images and accurate assessment of flow may be related to the size of the left atrium and angle of the pulmonary vein.
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Ecocardiografía Transesofágica/métodos , Venas Pulmonares/fisiopatología , Adulto , Anciano , Anciano de 80 o más Años , Análisis de Varianza , Velocidad del Flujo Sanguíneo , Ecocardiografía Doppler de Pulso/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Venas Pulmonares/diagnóstico por imagen , Flujo Sanguíneo RegionalRESUMEN
Prospective surveillance of 63 human immunodeficiency virus (HIV)-positive patients and 9 HIV-negative partners over 5-27 months yielded 51 adenoviruses from 18 HIV-positive patients. These were serotyped and compared by restriction enzyme analysis (REA) together with 24 isolates from 19 other HIV-positive patients. The actuarial risk of infection at 1 year in HIV-positive patients was 28% (17% with entry CD4 cell count of > 200/mm3 and 38% with CD4 cell count of < or = 200/mm3, P = .03). The most frequent site of infection was gastrointestinal (17/18 patients) with mainly subgenus D adenoviruses, while urinary infection was caused by subgenus B or D. Prolonged fecal excretion (2-27 months) was associated with CD4 cell counts < 150/mm3. Identical strains were seen in 2 HIV-positive partners and 2 unrelated patients. Gastrointestinal infection was temporally associated with diarrhea in only 7 (41%) of 17 cases. The remainder (59%) were asymptomatic or minimally symptomatic, and diarrhea was often caused by other opportunistic pathogens.
Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/epidemiología , Síndrome de Inmunodeficiencia Adquirida/fisiopatología , Infecciones por Adenovirus Humanos/epidemiología , Adenovirus Humanos/aislamiento & purificación , Seropositividad para VIH/complicaciones , Complejo Relacionado con el SIDA/fisiopatología , Análisis Actuarial , Adenovirus Humanos/clasificación , Adenovirus Humanos/genética , Adulto , Estudios de Casos y Controles , Femenino , Seronegatividad para VIH , Humanos , Estudios Longitudinales , Masculino , Prohibitinas , Mapeo Restrictivo , Factores de Riesgo , SerotipificaciónRESUMEN
STUDY DESIGN: This study analyzed the anatomic relationships between bony structures and soft tissues of the cervicothoracic junction. OBJECTIVES: To provide composite reference data for intrasegmental and intersegmental gradients of anatomic variation within the cervical-thoracic junction. SUMMARY OF BACKGROUND DATA: Because the risk of soft tissue damage during posterior spinal stabilization, an understanding of bony and soft tissue changes in the cervicothoracic junction is necessary. METHODS: Three-hundred-twenty-four cross-sectional spinal segments from nine spines were analyzed to characterize cervicothoracic junctional anatomy. RESULTS: There were predictable cranial-to-caudal alterations in both bone and soft tissue anatomy of the cervicothoracic junction. Neural and vascular structures directly anterior to the lateral mass or transverse process and lateral to the pedicle tend to decrease in frequency, whereas measured parameters of the vertebrae increase in size from C5-T3, except for pedicle dimensions that tend to increase at the C7-T1 junction. CONCLUSION: The anatomic changes that occur within the cervicothoracic junction are consistent and predictable, and their recognition should lead to a better appreciation of their clinical implications.