RESUMEN

INTRODUCTION: In the last decades sports practice in children has increased, thus increasing the number of musculoskeletal injuries. There are no validated scales in Spanish for the functional evaluation of knee aspects in children. The validation and cross-cultural adaptation of the Pedi-IKDC scale to Spanish, used for this purpose, was carried out. METHODOLOGY: The scale was applied to 50 patients that suffered traumatic knee injuries between 2016 and 2021 and underwent surgical interventions. The validation process of the scale was carried out after a pilot test, cross-cultural adaptation of words into Spanish, translation-retranslation, statistical, determination of internal consistency of the instrument, intraclass correlation, test-retest and evaluation of data dispersion. RESULTS: The internal consistency of the evaluated instrument is good according to the Gregory scale with a Cronbach's alpha of 0.82 The intraclass correlation was considered substantial (0.624) and the test-retest correlation, showed a coefficient of 0.91. The Bland-Altman graph showed a low dispersion among the data. CONCLUSION: The Pedi-IKDC scale can be a useful tool to assess functionality in children who have undergone knee surgery, it is considered valid, with adequate reliability and with the advantage of easy application.

RESUMEN

INTRODUCTION: In the last decades sports practice in children has increased, thus increasing the number of musculoskeletal injuries. There are no validated scales in Spanish for the functional evaluation of knee aspects in children. The validation and cross-cultural adaptation of the Pedi-IKDC scale to Spanish, used for this purpose, was carried out. METHODOLOGY: The scale was applied to 50 patients that suffered traumatic knee injuries between 2016 and 2021 and underwent surgical interventions. The validation process of the scale was carried out after a pilot test, cross-cultural adaptation of words into Spanish, translation-retranslation, statistical, determination of internal consistency of the instrument, intraclass correlation, test-retest and evaluation of data dispersion. RESULTS: The internal consistency of the evaluated instrument is good according to the Gregory scale with a Cronbach's alpha of 0.82 The intraclass correlation was considered substantial (0.624) and the test-retest correlation, showed a coefficient of 0.91. The Bland-Altman graph showed a low dispersion among the data. CONCLUSION: The Pedi-IKDC scale can be a useful tool to assess functionality in children who have undergone knee surgery, it is considered valid, with adequate reliability and with the advantage of easy application.

RESUMEN

Resumen Los tumores cerebrales son una causa importante de las epilepsias de difícil manejo, corresponden a un 20-30 % de los casos de cirugía de epilepsia refractaria. En este grupo de pacientes los tumores neuroepiteliales de bajo grado asociados a epilepsia (LEAT) son la principal causa, siendo los más frecuentes los tumores neuroepiteliales disembrioplásticos (DNT) y ganglioglioma (GG). En el presente artículo revisamos los cambios en la definición de epilepsia refractaria, avances en el diagnóstico por imágenes y el diagnóstico histopatológico con los nuevos marcadores moleculares, que han permitido un diagnóstico cada vez más precoz y certero. Se revisa también la cirugía resectiva que permite en estos casos una libertad de crisis cercana un 70-90% de los pacientes. Los mejores resultados en términos de control de crisis, se pueden alcanzar cuando la cirugía es precoz.

Brain tumors are an important cause of epilepsy that is difficult to manage, accounting for 20-30% of cases of refractory epilepsy surgery. In this group of patients, low-grade epilepsy-associated neuroepithelial tumors (LEAT) are the main cause and the most frequent being dysembryoplastic neuroepithelial tumors (DNT) and ganglioglioma (GG). In this article, we review the changes in the definition of refractory epilepsy, advances in diagnostic imaging, and histopathological diagnosis with new molecular markers, which have allowed for an increasingly early and accurate diagnosis. Resective surgery is also reviewed, allowing in these cases a seizure freedom close to 70-90% of patients. The best outcome in terms of seizure control can be achieved when early surgery is performed.

Asunto(s)

RESUMEN

Resumen: Introducción: La evaluación SANE (Single Assessment Numeric Evaluation) es un tipo de desenlace reportado por el paciente, corto y fácil de administrar que se correlaciona con otras evaluaciones funcionales del hombro más extensas. Hasta la fecha no se encuentra validada en nuestra lengua. Objetivo: Traducir, validar y adaptar la evaluación SANE al idioma español. Material y métodos: Se realizó la traducción y contra-traducción por dos evaluadores expertos bilingües y un traductor oficial. Se determinó la validez de constructo con un grupo de expertos en cirugía de hombro, la validez de criterio con la escala de American Shoulder and Elbow Surgeons (ASES) previamente validada en español y la reproducibilidad de la prueba. Resultados: Se evaluaron 113 pacientes con un promedio de edad de 53.6 años, siendo 56% mujeres. El diagnóstico más frecuente fue síndrome de manguito rotador en un 78.6% seguido por artrosis glenohumeral y acromioclavicular, hombro congelado e inestabilidad del hombro. El índice de correlación de Pearson entre el SANE y el ASES fue de 0.699. La reproducibilidad de la prueba fue alta con un coeficiente de correlación intraclase de 0.86. Conclusiones: La evaluación SANE es una medida de desenlace centrada en el paciente fiable y válida, que tiene una buena correlación con puntajes funcionales previamente validados al español más extensos. Se presenta la versión traducida y adaptada al español, que puede ser utilizada como medida de desenlace para intervenciones quirúrgicas o no quirúrgicas en patologías de hombro.

Abstract: Introduction: The Single Assessment Numeric Evaluation (SANE) score is a brief method of evaluating patients' perception of the shoulder joint and has proven high correlation with other outcome measures that are more lengthy and time-consuming in daily practice. The SANE score to date has not been validated in Spanish. Objective: To translate, adapt, and validate the SANE score in Spanish. Material and methods: An initial translation and counter-translation was conducted by two bilingual evaluators and an official translator. Content validity was evaluated by a group of experts in shoulder surgery. The criteria validity was determined by determining correlation with the previously translated and validated ASES score. Reliability of the test was determined. Results: 113 patients were evaluated, with an average age of 53.6 being 56% female. The most frequent diagnosis was rotator cuff syndrome in 78.6% followed by frozen shoulder, glenohumeral and acromioclavicular osteoarthritis and shoulder instability and acromioclavicular dislocation. Correlation between the SANE and ASES results was 0.699. The test was highly reliable with an intraclass correlation coefficient of 0.86. Conclusion: The SANE score is a valid and reliable patient centered outcome measure that has a good correlation with other previously validated scores in Spanish that are less practical. We present a translated valid version of the SANE score in Spanish that can be used as a patient reported outcome measure for shoulder pathologies.

RESUMEN

INTRODUCTION: The Single Assessment Numeric Evaluation (SANE) score is a brief method of evaluating patients' perception of the shoulder joint and has proven high correlation with other outcome measures that are more lengthy and time-consuming in daily practice. The SANE score to date has not been validated in Spanish. OBJECTIVE: To translate, adapt, and validate the SANE score in Spanish. MATERIAL AND METHODS: An initial translation and counter-translation was conducted by two bilingual evaluators and an official translator. Content validity was evaluated by a group of experts in shoulder surgery. The criteria validity was determined by determining correlation with the previously translated and validated ASES score. Reliability of the test was determined. RESULTS: 113 patients were evaluated, with an average age of 53.6 being 56% female. The most frequent diagnosis was rotator cuff syndrome in 78.6% followed by frozen shoulder, glenohumeral and acromioclavicular osteoarthritis and shoulder instability and acromioclavicular dislocation. Correlation between the SANE and ASES results was 0.699. The test was highly reliable with an intraclass correlation coefficient of 0.86. CONCLUSION: The SANE score is a valid and reliable patient centered outcome measure that has a good correlation with other previously validated scores in Spanish that are less practical. We present a translated valid version of the SANE score in Spanish that can be used as a patient reported outcome measure for shoulder pathologies.

INTRODUCCIÓN: La evaluación SANE (Single Assessment Numeric Evaluation) es un tipo de desenlace reportado por el paciente, corto y fácil de administrar que se correlaciona con otras evaluaciones funcionales del hombro más extensas. Hasta la fecha no se encuentra validada en nuestra lengua. OBJETIVO: Traducir, validar y adaptar la evaluación SANE al idioma español. MATERIAL Y MÉTODOS: Se realizó la traducción y contra-traducción por dos evaluadores expertos bilingües y un traductor oficial. Se determinó la validez de constructo con un grupo de expertos en cirugía de hombro, la validez de criterio con la escala de American Shoulder and Elbow Surgeons (ASES) previamente validada en español y la reproducibilidad de la prueba. RESULTADOS: Se evaluaron 113 pacientes con un promedio de edad de 53.6 años, siendo 56% mujeres. El diagnóstico más frecuente fue síndrome de manguito rotador en un 78.6% seguido por artrosis glenohumeral y acromioclavicular, hombro congelado e inestabilidad del hombro. El índice de correlación de Pearson entre el SANE y el ASES fue de 0.699. La reproducibilidad de la prueba fue alta con un coeficiente de correlación intraclase de 0.86. CONCLUSIONES: La evaluación SANE es una medida de desenlace centrada en el paciente fiable y válida, que tiene una buena correlación con puntajes funcionales previamente validados al español más extensos. Se presenta la versión traducida y adaptada al español, que puede ser utilizada como medida de desenlace para intervenciones quirúrgicas o no quirúrgicas en patologías de hombro.

Asunto(s)

RESUMEN

The current study used clinical and statistical significance tests to investigate the effects of two forms (didactic or interactive) of a universal prevention program on attitudes about shape and weight, eating behaviors, the influence of body aesthetic models, and self-esteem. Three schools were randomly assigned to one, interactive, didactic, or a control condition. Children (61 girls and 59 boys, age 9-11 years) were evaluated at pre-intervention, post-intervention, and at 6-month follow-up. Programs comprised eight, 90-min sessions. Statistical and clinical significance tests showed more changes in boys and girls with the interactive program versus the didactic intervention and control groups. The findings support the use of interactive programs that highlight identified risk factors and construction of identity based on positive traits distinct to physical appearance.

Asunto(s)

RESUMEN

Los trastornos de la audición presentan una alta prevalencia en la población infantil con graves consecuencias para la comunicación, y para el desarrollo cognitivo y emocional del niño. En los últimos años se han generado importantes avances en las tecnologías disponibles para la detección temprana de las pérdidas auditivas, lo cual ha permitido la creación de programas de cribado auditivo en numerosos países según sus propias características organizativas y tecnológicas. Objetivo. Se describe y fundamenta el modelo adoptado por el programa cubano de cribado auditivo, con múltiples grupos de riesgo, en funcionamiento desde 1983. Desarrollo. Se presentan datos acerca de la prevalencia de los trastornos auditivos, edad media de detección, cobertura del cribado, programas de estimulación temprana utilizados y sus efectos sobre el desarrollo cognitivo y progreso escolar del niño. Finalmente, se discuten los resultados de la introducción de una nueva tecnología diagnóstica para la evaluación objetiva de la audición en toda la gama de frecuencias audibles, basada en el registro de potenciales evocados auditivos de estado estable (PEAee) mediante el sistema cubano AUDIX. Conclusiones. El modelo cubano de cribado auditivo puede considerarse una alternativa válida para la detección temprana de trastornos auditivos en condiciones de limitados recursos. Durante su período de funcionamiento óptimo logró reducir a 10 meses la edad media de detección. El grupo de niños identificados e intervenidos tempranamente logró un mejor desarrollo cognitivo, afectivo y lingüístico. El uso de los PEAee a múltiples frecuencias en un protocolo de cribado auditivo puede aportar valiosa información audiométrica(AU)

Asunto(s)

RESUMEN

Introduction. Infant hearing loss is a highly prevalent disorder which untreated can severely disrupt normal brain development. As a result there is a significant delay in language acquisition as well as many cognitive and emotional problems in the child. Over the last decades important advances have occurred in the available technology for early detection and assessment of hearing impairment. Therefore many countries worldwide have become aware of the need for hearing screening programs. The optimal protocols depending on the local conditions of health care as well as the availability of technological and human resources. Aim. To summarize the results obtained over the last 20 years by an ongoing hearing screening protocol. Development. Data on the coverage program, sensitivity and specificity, age of identification of hearing losses, diagnostic and intervention stages will be summarized and discussed. Also the long terms effects of early detection on the child cognitive and language development are analyzed. Finally, the possible role of a new technique based on the recording of multiple auditory steady state potentials with Cuban equipment (AUDIX system) was evaluated within this context. Conclusions. The Cuban targeting multiple high risk hearing screening program is a useful alternative to early detection of hearing losses. The average detection age of hearing loss was 10 months during the period of optimal functioning. Cognitive, emotional and linguistic development are improved by early detection and intervention. The multiple auditory steady state responses can provide valuable audiometric information within a screening context (AU)

Introducción. Los trastornos de la audición presentan una alta prevalencia en la población infantil con graves consecuencias para la comunicación, y para el desarrollo cognitivo y emocional del niño. En los últimos años se han generado importantes avances en las tecnologías disponibles para la detección temprana de las pérdidas auditivas, lo cual ha permitido la creación de programas de cribado auditivo en numerosos países según sus propias características organizativas y tecnológicas. Objetivo. Se describe y fundamenta el modelo adoptado por el programa cubano de cribado auditivo, con múltiples grupos de riesgo, en funcionamiento desde 1983. Desarrollo. Se presentan datos acerca de la prevalencia de los trastornos auditivos, edad media de detección, cobertura del cribado, programas de estimulación temprana utilizados y sus efectos sobre el desarrollo cognitivo y progreso escolar del niño. Finalmente, se discuten los resultados de la introducción de una nueva tecnología diagnóstica para la evaluación objetiva de la audición en toda la gama de frecuencias audibles, basada en el registro de potenciales evocados auditivos de estado estable (PEAee) mediante el sistema cubano AUDIX. Conclusiones. El modelo cubano de cribado auditivo puede considerarse una alternativa válida para la detección temprana de trastornos auditivos en condiciones de limitados recursos. Durante su período de funcionamiento óptimo logró reducir a 10 meses la edad media de detección. El grupo de niños identificados e intervenidos tempranamente logró un mejor desarrollo cognitivo, afectivo y lingüístico. El uso de los PEAee a múltiples frecuencias en un protocolo de cribado auditivo puede aportar valiosa información audiométrica (AU)

Asunto(s)

RESUMEN

INTRODUCTION: Infant hearing loss is a highly prevalent disorder which untreated can severely disrupt normal brain development. As a result there is a significant delay in language acquisition as well as many cognitive and emotional problems in the child. Over the last decades important advances have occurred in the available technology for early detection and assessment of hearing impairment. Therefore many countries worldwide have become aware of the need for hearing screening programs. The optimal protocols depending on the local conditions of health care as well as the availability of technological and human resources. AIM: To summarize the results obtained over the last 20 years by an ongoing hearing screening protocol. DEVELOPMENT: Data on the coverage program, sensitivity and specificity, age of identification of hearing losses, diagnostic and intervention stages will be summarized and discussed. Also the long terms effects of early detection on the child cognitive and language development are analyzed. Finally, the possible role of a new technique based on the recording of multiple auditory steady state potentials with Cuban equipment (AUDIX system) was evaluated within this context. CONCLUSIONS: The Cuban targeting multiple high risk hearing screening program is a useful alternative to early detection of hearing losses. The average detection age of hearing loss was 10 months during the period of optimal functioning. Cognitive, emotional and linguistic development are improved by early detection and intervention. The multiple auditory steady state responses can provide valuable audiometric information within a screening context.

Asunto(s)

RESUMEN

Homo neanderthalensis has a unique combination of craniofacial features that are distinct from fossil and extant 'anatomically modern' Homo sapiens (modern humans). Morphological evidence, direct isotopic dates and fossil mitochondrial DNA from three Neanderthals indicate that the Neanderthals were a separate evolutionary lineage for at least 500,000 yr. However, it is unknown when and how Neanderthal craniofacial autapomorphies (unique, derived characters) emerged during ontogeny. Here we use computerized fossil reconstruction and geometric morphometrics to show that characteristic differences in cranial and mandibular shape between Neanderthals and modern humans arose very early during development, possibly prenatally, and were maintained throughout postnatal ontogeny. Postnatal differences in cranial ontogeny between the two taxa are characterized primarily by heterochronic modifications of a common spatial pattern of development. Evidence for early ontogenetic divergence together with evolutionary stasis of taxon-specific patterns of ontogeny is consistent with separation of Neanderthals and modern humans at the species level.

Asunto(s)

RESUMEN

During virus entry, herpes simplex virus (HSV) glycoprotein D (gD) binds to one of several human cellular receptors. One of these, herpesvirus entry mediator A (HveA), is a member of the tumor necrosis factor receptor (TNFR) superfamily, and its ectodomain contains four characteristic cysteine-rich pseudorepeat (CRP) elements. We previously showed that gD binds the ectodomain of HveA expressed as a truncated, soluble protein [HveA(200t)]. To localize the gD-binding domain of HveA, we expressed three additional soluble forms of HveA consisting of the first CRP [HveA(76t)], the second CRP [HveA(77-120t)], or the first and second CRPs [HveA(120t)]. Biosensor and enzyme-linked immunosorbent assay studies showed that gD bound to HveA(120t) and HveA(200t) with the same affinity. However, gD did not bind to HveA(76t) or HveA(77-120t). Furthermore, HveA(200t) and HveA(120t), but not HveA(76t) or HveA(77-120t), blocked herpes simplex virus (HSV) entry into CHO cells expressing HveA. We also generated six monoclonal antibodies (MAbs) against HveA(200t). MAbs CW1, -2, and -4 bound linear epitopes within the second CRP, while CW7 and -8 bound linear epitopes within the third or fourth CRPs. None of these MAbs blocked the binding of gD to HveA. In contrast, MAb CW3 recognized a discontinuous epitope within the first CRP of HveA, blocked the binding of gD to HveA, and exhibited a limited ability to block virus entry into cells expressing HveA, suggesting that the first domain of HveA contains at least a portion of the gD binding site. The inability of gD to bind HveA(76t) suggests that additional amino acid residues of the gD binding site may reside within the second CRP.

Asunto(s)

RESUMEN

The human herpesvirus entry mediator C (HveC), also known as the poliovirus receptor-related protein 1 (PRR1) and as nectin-1, allows the entry of herpes simplex virus type 1 (HSV-1) and HSV-2 into mammalian cells. The interaction of virus envelope glycoprotein D (gD) with such a receptor is an essential step in the process leading to membrane fusion. HveC is a member of the immunoglobulin (Ig) superfamily and contains three Ig-like domains in its extracellular portion. The gD binding site is located within the first Ig-like domain (V domain) of HveC. We generated a panel of monoclonal antibodies (MAbs) against the ectodomain of HveC. Eleven of these, which detect linear or conformational epitopes within the V domain, were used to map a gD binding site. They allowed the detection of HveC by enzyme-linked immunosorbent assay, Western blotting, and biosensor analysis or directly on the surface of HeLa cells and human neuroblastoma cell lines, as well as simian Vero cells. The anti-HveC V-domain MAbs CK6, CK8, and CK41, as well as the previously described MAb R1.302, blocked HSV entry. Their binding to soluble HveC was blocked by the association of gD with the receptor, indicating that their epitopes overlap a gD binding site. Competition assays on an optical biosensor showed that CK6 and CK8 (linear epitopes) inhibited the binding of CK41 and R1.302 (conformational epitopes) to HveC and vice versa. Epitope mapping showed that CK6 and CK8 bound between residues 80 and 104 of HveC, suggesting that part of the gD binding site colocalizes in the same region.

Asunto(s)

Asunto(s)

RESUMEN

Mediante el inventario de evaluación sussmilch (IES), previamente validado y confiabilizado, se estableció una diferencia significativa entre la teoría y la realidad de la calidad de la información publicada (1967-1999). Esto señala que esos resultados no pueden generalizarse ni dentro ni fuera de las muestras de estudiantes, también de medicina. En contraste, con base en el principio de que resultados negativos también son resultados, en este ejercicio de demostró empíricamente que los resultados obtenidos representan bien a las muestras de estudiantes que fueron admitidos a cinco facultades de medicina hispanoamericana (FMH). Se desarrolló el indice de alcoholismo de pregrado (IAPG) integrado por cuatro (ó cinco) reactivos con respuestas dicotómicas, capaces de discernir, con error conocido, entre estudiantes de medicina no-alcohólicos (NOH), sospechosos (SOH) y alcohólicos (OH). El perfil psicométrico del IAPG exhibió una equivalente entre las Facultades de Medicina de la Universidad Nacional Autónoma de México (UNAM), Universidad Nacional de Colombia (UNC), Universidad Peruana Cayetano Heredia(UPCH), universidad Nacional de San Antonio Abad del Cuzco (UNSAAC) y Universidad de Cuenca(UC).

Asunto(s)

RESUMEN

The human herpesvirus entry mediator C (HveC/PRR1) is a member of the immunoglobulin family used as a cellular receptor by the alphaherpesviruses herpes simplex virus (HSV), pseudorabies virus, and bovine herpesvirus type 1. We previously demonstrated direct binding of the purified HveC ectodomain to purified HSV type 1 (HSV-1) and HSV-2 glycoprotein D (gD). Here, using a baculovirus expression system, we constructed and purified truncated forms of the receptor containing one [HveC(143t)], two [HveC(245t)], or all three immunoglobulin-like domains [HveC(346t)] of the extracellular region. All three constructs were equally able to compete with HveC(346t) for gD binding. The variable domain bound to virions and blocked HSV infection as well as HveC(346t). Thus, all of the binding to the receptor occurs within the first immunoglobulin-like domain, or V-domain, of HveC. These data confirm and extend those of Cocchi et al. (F. Cocchi, M. Lopez, L. Menotti, M. Aoubala, P. Dubreuil, and G. Campadelli-Fiume, Proc. Natl. Acad. Sci. USA 95:15700, 1998). Using biosensor analysis, we measured the affinity of binding of gD from HSV strains KOS and rid1 to two forms of HveC. Soluble gDs from the KOS strain of HSV-1 had the same affinity for HveC(346t) and HveC(143t). The mutant gD(rid1t) had an increased affinity for HveC(346t) and HveC(143t) due to a faster rate of complex formation. Interestingly, we found that HveC(346t) was a tetramer in solution, whereas HveC(143t) and HveC(245t) formed dimers, suggesting a role for the third immunoglobulin-like domain of HveC in oligomerization. In addition, the stoichiometry between gD and HveC appeared to be influenced by the level of HveC oligomerization.

Asunto(s)

RESUMEN

In this study, we present a new computerized reconstruction of the Le Moustier 1 Neanderthal skull and discuss its significance for Neanderthal growth and variability. Because of the precarious state of preservation of the original material, we applied entirely noninvasive methods of fossil reconstruction and morphometry, using a combination of computed tomography, computer graphics, and stereolithography. After electronic restoration, the isolated original pieces were recomposed on the computer screen using external and internal anatomical clues to position the bone fragments and mirror images to complete missing parts. The inferred effects of general compressive deformation that occurred during fossilization were corrected by virtual decompression of the skull. The resulting new reconstruction of the Le Moustier 1 skull shows morphologic features close to the typical Neanderthal adult state. Residual asymmetry of skeletal parts can be traced to in vivo skeletal modification: the left mandibular joint shows signs of a healed condylar fracture, and the anatomy of the occipital region suggests mild plagiocephaly. Using micro-CT analysis, the left incus could be recovered from the matrix filling of the middle ear cavity. Its morphometric dimensions are similar to those of the La Ferrassie III incus. The morphometric characteristics of the inner ear deviate substantially from the condition reported as typical for Neanderthals and fall within the range of modern human variability.

Asunto(s)

RESUMEN

BACKGROUND: Medical devices that are used on patients in fields containing potentially infectious body fluids can become contaminated and transmit infectious agents to other sites on the patient or to other patients if the devices are not properly cleaned and decontaminated after use on each patient treatment site. One such device is the needleless or jet injector, which is widely used in medicine and dentistry to deliver local anesthetic in procedures such as bone marrow aspirations, lumbar punctures, and cutaneous and intraoral injections. This study was conducted to determine whether cross-contamination can occur on in vitro reuse of a needleless injector and whether a manufacturer's recommended method of injector decontamination (ie, immersion sterilization) is effective in the prevention of cross-contamination. METHODS: The study was performed with new autoclaved injectors, fluorescein dye, and Streptococcus crista (the bacteria commonly found in saliva) in the field of use to determine whether these devices can become contaminated during use and carry over the contamination to other sites during immediate reuse. RESULTS: Fluorescein dye and bacteria tests with the needleless injectors showed that contamination or carryover does occur. It appeared to reduced to a minimum when a autoclaved, sterile rubber cap used over the head of the device during injection was replaced between each use, although replacement of the rubber cap alone did not prevent carryover. Immersion of the head of the injector in a 2% glutaraldehyde solution for 30 minutes followed by a sterile water rinse and the replacement of the rubber cap with a sterile cap between uses was shown to curtail bacterial growth and prevent cross-contamination on immediate reuse of the device. CONCLUSION: This study demonstrated that needleless injectors become contaminated during in vitro use and direct contact with contaminated surfaces and that needless injectors carry over the contamination to subsequent sites of release. The replacement of the injector's rubber cap with a new one after initial discharge or the removal of an exposed rubber cap and immersion of the head of the injector in 2% glutaraldehyde followed by a rinse of the head in sterile water, as recommended by one injector manufacturer, can minimize or eliminate the carryover.

Asunto(s)

RESUMEN

The herpes simplex virus (HSV) gH-gL complex is essential for virus infectivity and is a major antigen for the host immune system. The association of gH with gL is required for correct folding, cell surface trafficking, and membrane presentation of the complex. Previously, a mammalian cell line was constructed which produces a secreted form of gHt-gL complex lacking the transmembrane and cytoplasmic tail regions of gH. gHt-gL retains a conformation similar to that of its full-length counterpart in HSV-infected cells. Here, we examined the structural and antigenic properties of gHt-gL. We first determined its stoichiometry and carbohydrate composition. We found that the complex consists of one molecule each of gH and gL. The N-linked carbohydrate (N-CHO) site on gL and most of the N-CHO sites on gH are utilized, and both proteins also contain O-linked carbohydrate and sialic acid. These results suggest that the complex is processed to the mature form via the Golgi network prior to secretion. To determine the antigenically active sites of gH and gL, we mapped the epitopes of a panel of gH and gL monoclonal antibodies (MAbs), using a series of gH and gL C-terminal truncation variant proteins produced in transiently transfected mammalian cells. Sixteen gH MAbs (including H6 and 37S) reacted with the N-terminal portion of gH between amino acids 19 and 276. One of the gH MAbs, H12, reacted with the middle portion of gH (residues 476 to 678). Nine gL MAbs (including 8H4 and VIII 62) reacted with continuous epitopes within the C-terminal portion of gL, and this region was further mapped within amino acids 168 to 178 with overlapping synthetic peptides. Finally, plasmids expressing the gH and gL truncations were employed in cotransfection assays to define the minimal regions of both gH and gL required for complex formation and secretion. The first 323 amino acids of gH and the first 161 amino acids of gL can form a stable secreted hetero-oligomer with gL and gH792, respectively, while gH323-gL168 is the smallest secreted hetero-oligomer. The first 648 amino acids of gH are required for reactivity with MAbs LP11 and 53S, indicating that a complex of gH648-gL oligomerizes into the correct conformation. The data suggest that both antigenic activity and oligomeric structure require the amino-terminal portions of gH and gL.

Asunto(s)

RESUMEN

HVEM (for herpesvirus entry mediator) is a member of the tumor necrosis factor receptor superfamily and mediates entry of many strains of herpes simplex virus (HSV) into normally nonpermissive Chinese hamster ovary (CHO) cells. We used sucrose density centrifugation to demonstrate that purified HSV-1 KOS virions bind directly to a soluble, truncated form of HVEM (HVEMt) in the absence of any other cell-associated components. Therefore, HVEM mediates HSV entry by serving as a receptor for the virus. We previously showed that soluble, truncated forms of HSV glycoprotein D (gDt) bind to HVEMt in vitro. Here we show that antibodies specific for gD, but not the other entry glycoproteins gB, gC, or the gH/gL complex, completely block HSV binding to HVEM. Thus, virion gD is the principal mediator of HSV binding to HVEM. To map sites on virion gD which are necessary for its interaction with HVEM, we preincubated virions with gD-specific monoclonal antibodies (MAbs). MAbs that recognize antigenic sites Ib and VII of gD were the only MAbs which blocked the HSV-HVEM interaction. MAbs from these two groups failed to coprecipitate HVEMt in the presence of soluble gDt, whereas the other anti-gD MAbs coprecipitated HVEMt and gDt. Previous mapping data indicated that site VII includes amino acids 11 to 19 and site Ib includes 222 to 252. The current experiments indicate that these sites contain residues important for HSV binding to HVEM. Group Ib and VII MAbs also blocked HSV entry into HVEM-expressing CHO cells. These results suggest that the mechanism of neutralization by these MAbs is via interference with the interaction between gD in the virus and HVEM on the cell. Group Ia and II MAbs failed to block HSV binding to HVEM yet still neutralized HVEM-mediated entry, suggesting that these MAbs block entry at a step other than HVEM binding.

Asunto(s)

RESUMEN

The herpes simplex virus type 1 (HSV-1) gH-gL complex which is found in the virion envelope is essential for virus infectivity and is a major antigen for the host immune system. However, little is known about the precise role of gH-gL in virus entry, and attempts to demonstrate the immunologic or vaccine efficacy of gH and gL separately or as the gH-gL complex have not succeeded. We constructed a recombinant mammalian cell line (HL-7) which secretes a soluble gH-gL complex, consisting of gH truncated at amino acid 792 (gHt) and full-length gL. Purified gHt-gL reacted with gH- and gL-specific monoclonal antibodies, including LP11, which indicates that it retains its proper antigenic structure. Soluble forms of gD (gDt) block HSV infection by interacting with specific cellular receptors. Unlike soluble gD, gHt-gL did not block HSV-1 entry into cells, nor did it enhance the blocking capacity of gD. However, polyclonal antibodies to the complex did block entry even when added after virus attachment. In addition, these antibodies exhibited high titers of complement-independent neutralizing activity against HSV-1. These sera also cross-neutralized HSV-2, albeit at low titers, and cross-reacted with gH-2 present in extracts of HSV-2-infected cells. To test the potential for gHt-gL to function as a vaccine, BALB/c mice were immunized with the complex. As controls, other mice were immunized with gD purified from HSV-infected cells or were sham immunized. Sera from the gD- or gHt-gL-immunized mice exhibited high titers of virus neutralizing activity. Using a zosteriform model of infection, we challenged mice with HSV-1. All animals showed some evidence of infection at the site of virus challenge. Mice immunized with either gD or gHt-gL showed reduced primary lesions and exhibited no secondary zosteriform lesions. The sham-immunized control animals exhibited extensive secondary lesions. Furthermore, mice immunized with either gD or gHt-gL survived virus challenge, while many control animals died. These results suggest that gHt-gL is biologically active and may be a candidate for use as a subunit vaccine.

Asunto(s)