A Pilot Study on the Feasibility of an Extended Suckling System for Pasture-Based Dairies.

This study investigated cow-calf productivity in a 10-week, pasture-based, extended suckling system featuring part-time cow-calf contact and once-a-day milking. A total of 30 dairy cows and their calves were assigned to two treatments: (1) cow and calf managed in an extended suckling system; or (2) cow and calf separated at birth and managed as usual. Cow-calf pairs grazed together during the day and spent the night separated by fence-line contact. The dams were reunited with the calves after once-a-day milking every morning. The commercial treatment pairs were separated after birth, and cows were milked twice a day and managed within the farm herd. Commercial calves were reared and managed as per commercial Australian practices. Cow-calf dams yielded 9 L/cow/day less saleable milk (p < 0.001), and their milk had lower fat (p = 0.04) but a higher protein percentage (p < 0.001) than commercial cows during pre-weaning. However, milk yield and composition were comparable post-weaning. Dam-suckled calves gained weight faster and were therefore weaned 2 weeks earlier than commercial calves, which were offered 8 L/day milk. This study has demonstrated a novel system of extended cow-calf suckling that could be practical to implement in pasture-based dairies. The long-term effects and scalability of the extended suckling system described here require further validation.

Are indigenous territories effective natural climate solutions? A neotropical analysis using matching methods and geographic discontinuity designs.

Indigenous Territories (ITs) with less centralized forest governance than Protected Areas (PAs) may represent cost-effective natural climate solutions to meet the Paris agreement. However, the literature has been limited to examining the effect of ITs on deforestation, despite the influence of anthropogenic degradation. Thus, little is known about the temporal and spatial effect of allocating ITs on carbon stocks dynamics that account for losses from deforestation and degradation. Using Amazon Basin countries and Panama, this study aims to estimate the temporal and spatial effects of ITs and PAs on carbon stocks. To estimate the temporal effects, we use annual carbon density maps, matching analysis, and linear mixed models. Furthermore, we explore the spatial heterogeneity of these estimates through geographic discontinuity designs, allowing us to assess the spatial effect of ITs and PAs boundaries on carbon stocks. The temporal effects highlight that allocating ITs preserves carbon stocks and buffer losses as well as allocating PAs in Panama and Amazon Basin countries. The geographic discontinuity designs reveal that ITs' boundaries secure more extensive carbon stocks than their surroundings, and this difference tends to increase towards the least accessible areas, suggesting that indigenous land use in neotropical forests may have a temporarily and spatially stable impact on carbon stocks. Our findings imply that ITs in neotropical forests support Nationally Determined Contributions (NDCs) under the Paris Agreement. Thus, Indigenous peoples must become recipients of countries' results-based payments.

The role of forest conversion, degradation, and disturbance in the carbon dynamics of Amazon indigenous territories and protected areas.

Maintaining the abundance of carbon stored aboveground in Amazon forests is central to any comprehensive climate stabilization strategy. Growing evidence points to indigenous peoples and local communities (IPLCs) as buffers against large-scale carbon emissions across a nine-nation network of indigenous territories (ITs) and protected natural areas (PNAs). Previous studies have demonstrated a link between indigenous land management and avoided deforestation, yet few have accounted for forest degradation and natural disturbances-processes that occur without forest clearing but are increasingly important drivers of biomass loss. Here we provide a comprehensive accounting of aboveground carbon dynamics inside and outside Amazon protected lands. Using published data on changes in aboveground carbon density and forest cover, we track gains and losses in carbon density from forest conversion and degradation/disturbance. We find that ITs and PNAs stored more than one-half (58%; 41,991 MtC) of the region's carbon in 2016 but were responsible for just 10% (-130 MtC) of the net change (-1,290 MtC). Nevertheless, nearly one-half billion tons of carbon were lost from both ITs and PNAs (-434 MtC and -423 MtC, respectively), with degradation/disturbance accounting for >75% of the losses in 7 countries. With deforestation increasing, and degradation/disturbance a neglected but significant source of region-wide emissions (47%), our results suggest that sustained support for IPLC stewardship of Amazon forests is critical. IPLCs provide a global environmental service that merits increased political protection and financial support, particularly if Amazon Basin countries are to achieve their commitments under the Paris Climate Agreement.

Trombolisis intravenosa en ataque cerebrovascular isquémico agudo en Santa Cruz Bolivia: análisis retrospectivo de los primeros 18 casos / Intravenous thrombolysis in acute ischemic stroke in Santa Cruz Bolivia: retrospective analysis of the first 18 cases

OBJETIVOS: describir la experiencia en la implementación del protocolo de trombolisis i.v. en 18 pacientes con Accidente Cerebrovascular (ACV) isquémico en la Unidad de ACV de la Clínica Foianini, concretamente, entre septiembre de 2013 y julio de 2017. MÉTODOS: el estudio tiene un enfoque cuantitativo, tipo descriptivo y retrospectivo. Se revisó las historias clínicas de los pacientes con diagnóstico de ACV isquémico agudo, que fueron tratados con trombolitico intravenoso (Alteplasa) en el servicio UTAC, entre septiembre de 2013 y julio 2017 en Santa Cruz, Bolivia. RESULTADOS: el Tiempo Síntoma-Puerta alcanzo a 62,7 +/- 38 min, mientras que el Tiempo Puerta-Aguja fue de 53,6 +/- 15 min. y Tiempo Síntoma-Aguja 114,6 +/- 43 min. Se registraron complicaciones en 6 (33,3%). promedio de 4,6 +/- 3 días de internación en la clínica, pasando un promedio de 2,8 +/- 2 días en la Unidad de ACV. La tasa de mortalidad fue de 16,6%. Según la Escala Modficada de Rankin, tuvieron un puntaje de 0-1 mRS. CONCLUSIONES: el tiempo Puerta-Aguja en nuestro establecimiento fue de un promedio de 57 minutos, cumpliendo de esta forma la recomendación de la American Heart Association Guidelines 2013, la cual recalca que éste debe ser <60 min13.

OBJECTIVE: describe the experience in the implementation of the thrombolysis intravenous protocol in eighteen patients with acute ischemic stroke in the Stroke unit from the Foianini Clinic specifically, between September of 2013 and July of 2017. METHODS: the study has a quantitative, descriptive and retrospective approach. The medical record of the patients with the acute ischemic stroke diagnosis treated with intravenous thrombolytic (Alteplasa) in the UTAC service between September of 2013 and July 2017 in Santa Cruz Bolivia was revised. RESULTS: symptoms-Door time reached to 62.7 +/- 38 minutes. While the time Door- needle time was 53.6 +/- 15 minutes and the time symptoms- needle 114.6 +/- 3 days of internment in the clinic, passing out an average of 2.8 +/- 2 days in the stroke unit. The mortality tax was 16.6%. According to the modified Rankin Scale, it had a 0-1 mRS score. CONCLUSIONS: the time Door-needle in our establishment was an average of 57 minutes, fulfilling in this way the recommendation of the American Heart Association Guidelines 2013, which emphasize that it has to be < 60 minutes.

Medidas declaradas por padres y cuidadores de lactantes y preescolares para prevenir la exposición a humo de tabaco ambiental / Measures stated by parents and caregivers of infants, toddlers, and preschoolers to prevent environmental tobacco smoke exposure

Introducción. La exposición a humo de tabaco ambiental constituye un problema de salud pública por sus potenciales riesgos en los niños. Objetivo. Determinar las acciones preventivas adoptadas por padres y/o cuidadores de lactantes y preescolares respecto de la exposición a humo de tabaco ambiental. Métodos. Estudio transversal analítico en padres y cuidadores de lactantes y preescolares escolarizados, residentes en la ciudad de Valdivia, Chile. Se aplicó un cuestionario sobre medidas preventivas del humo de tabaco ambiental. Se analizó con estadística descriptiva y de asociación usando el test exacto de Fisher y la regresión logística para calcular odds ratio (OR) y sus respectivos intervalos de confianza del 95% (IC95%) como estimación de riesgo de no adopción de medidas preventivas en el hogar. Resultados. Participaron 469 padres y cuidadores. La prevalencia de hogares con un miembro fumador fue 51,8%. La mayoría fumaba fuera del hogar (92,2%). La medida más adoptada fuera del hogar fue evitar lugares con gente fumando. En este ambiente, hubo diferencia significativa entre hombres y mujeres en no prevenir la exposición (7,6% vs. 2,2%; p= 0,040). Se constató mayor riesgo de no prevenir el consumo en el hogar (dentro o fuera) cuando hubo algún miembro fumador (OR 3,55; IC95%: 2,05-6,14). Y menor riesgo cuando hubo dos o más niños en el hogar (OR 0,46; IC95%: 0,26-0,83). Conclusiones. La mayoría de los padres y cuidadores piden que se fume fuera del hogar y evitan los lugares con gente fumando, aunque, en los hogares con miembros fumadores, la prevención es menor.

Introduction. The exposure to environmental tobacco smoke is a public health problem because of its potential risks for children. Objective. To determine the preventive actions taken by parents and/or caregivers of infants, toddlers, and preschoolers regarding environmental tobacco smoke exposure. Methods. Cross-sectional, analytical study in parents and caregivers of infants, toddlers, and preschoolers from Valdivia, Chile. A questionnaire on preventive measures regarding environmental tobacco smoke was administered. The questionnaire was analyzed with descriptive and association statistics using Fisher's exact test and logistic regression to calculate the odds ratios (ORs) and their corresponding 95% confidence intervals (95% CIs) as an estimation of the risk for not taking preventive measures at home. Results. A total of 469 parents and caregivers participated. The prevalence of households with one member who smoked was 51.8%. Most of them smoked outside the house (92.2%). The most common measure taken outside the household was to avoid places where people smoked. In this setting, a significant difference was observed between men and women in relation to avoiding exposure (7.6% versus 2.2%; p= 0.040). A higher risk for not avoiding tobacco use inside or outside the housewas confirmed when a household member smoked (OR: 3.55; 95% CI: 2.05-6.14). Also, a lower risk was observed when there were two or more children in the household (OR: 0.46; 95% CI: 0.26-0.83). Conclusions. Most parents and caregivers ask others to smoke outside and avoid places where people smoke; however, the level of prevention is lower among households with smokers.

Measures stated by parents and caregivers of infants, toddlers, and preschoolers to prevent environmental tobacco smoke exposure. / Medidas declaradas por padres y cuidadores de lactantes y preescolares para prevenir la exposición a humo de tabaco ambiental.

INTRODUCTION: The exposure to environmental tobacco smoke is a public health problem because of its potential risks for children. Objective. To determine the preventive actions taken by parents and/or caregivers of infants, toddlers, and preschoolers regarding environmental tobacco smoke exposure. Methods. Cross-sectional, analytical study in parents and caregivers of infants, toddlers, and preschoolers from Valdivia, Chile. A questionnaire on preventive measures regarding environmental tobacco smoke was administered. The questionnaire was analyzed with descriptive and association statistics using Fisher's exact test and logistic regression to calculate the odds ratios (ORs) and their corresponding 95% confidence intervals (95% CIs) as an estimation of the risk for not taking preventive measures at home. RESULTS: A total of 469 parents and caregivers participated. The prevalence of households with one member who smoked was 51.8%. Most of them smoked outside the house (92.2%). The most common measure taken outside the household was to avoid places where people smoked. In this setting, a significant difference was observed between men and women in relation to avoiding exposure (7.6% versus 2.2%; p= 0.040). A higher risk for not avoiding tobacco use inside or outside the housewas confirmed when a household member smoked (OR: 3.55; 95% CI: 2.05-6.14). Also, a lower risk was observed when there were two or more children in the household (OR: 0.46; 95% CI: 0.26-0.83). CONCLUSIONS: Most parents and caregivers ask others to smoke outside and avoid places where people smoke; however, the level of prevention is lower among households with smokers.

Introducción. La exposición a humo de tabaco ambiental constituye un problema de salud pública por sus potenciales riesgos en los niños. Objetivo. Determinar las acciones preventivas adoptadas por padres y/o cuidadores de lactantes y preescolares respecto de la exposición a humo de tabaco ambiental. Métodos. Estudio transversal analítico en padres y cuidadores de lactantes y preescolares escolarizados, residentes en la ciudad de Valdivia, Chile. Se aplicó un cuestionario sobre medidas preventivas del humo de tabaco ambiental. Se analizó con estadística descriptiva y de asociación usando el test exacto de Fisher y la regresión logística para calcular odds ratio (OR) y sus respectivos intervalos de confianza del 95% (IC95%) como estimación de riesgo de no adopción de medidas preventivas en el hogar. Resultados. Participaron 469 padres y cuidadores. La prevalencia de hogares con un miembro fumador fue 51,8%. La mayoría fumaba fuera del hogar (92,2%). La medida más adoptada fuera del hogar fue evitar lugares con gente fumando. En este ambiente, hubo diferencia significativa entre hombres y mujeres en no prevenir la exposición (7,6% vs. 2,2%; p= 0,040). Se constató mayor riesgo de no prevenir el consumo en el hogar (dentro o fuera) cuando hubo algún miembro fumador (OR 3,55; IC95%: 2,05-6,14). Y menor riesgo cuando hubo dos o más niños en el hogar (OR 0,46; IC95%: 0,26-0,83). Conclusiones. La mayoría de los padres y cuidadores piden que se fume fuera del hogar y evitan los lugares con gente fumando, aunque, en los hogares con miembros fumadores, la prevención es menor.

Health benefits of hard martial arts in adults: a systematic review.

Participation in organized sports is promoted as a means of increasing physical activity levels and reducing chronic disease risk in adults. Hard martial arts practice (i.e. using body contact techniques), has gained in popularity over time. This review explores the evidence for health benefits of "hard" martial arts practice within the adult population. A systematic electronic database search was conducted, and quality assessments applied the Effective Public Health Practice Project tool. Twenty-eight studies met the inclusion criteria, examining balance, cognitive function, muscular skeletal status, psychological, cardiovascular fitness, and metabolic effects. The majority of studies reported positive effects resulting from hard martial arts practice, showing some improvement and maintenance of balance, cognitive function and psychological health. Benefits may be obtained regardless of the age of practice commencement. However, quality of the evidence is affected by methodological weaknesses across the studies. "Hard" martial arts seem to have potential to improve balance and cognitive functions that decline with age, which can lead to poorer health outcomes among the elderly (e.g. cognitive decline, falls and fractures). Benefits should be further investigated with improved intervention studies, representative samples and longer follow-up periods in order to establish associations with morbidity and mortality in the long term.

Characterization of a novel modification of a CHO-produced mAb: Evidence for the presence of tyrosine sulfation.

Herein we describe the investigation of a Chinese hamster ovary (CHO)-expressed human mAb molecule found partially modified by a +80 Da adduct. This mass difference, suggestive of a single sulfation or phosphorylation addition, was observed by mass analysis of the intact and reduced molecule by mass spectrometry (MS). The modification was located on tyrosine 31 (Y31) of the light chain in the complementarity-determining region 1 by liquid chromatography (LC)-MS peptide mapping and electron transfer dissociation fragmentation. The complete loss of the 80 Da modification moiety during collision induced dissociation fragmentation suggested this modification could not be a tyrosine phosphorylation. Treatment of the mAb with alkaline phosphatase confirmed our hypothesis. Western blot experiment using anti-tyrosine sulfation antibody and LC retention time correlation with corresponding synthetic sulfated peptides further confirmed the identification of tyrosine sulfation on the light chain. The unique sequence motif with neighboring acidic amino acids and local secondary structure might play a role to make Y31 a substrate residue for sulfation. This type of modification, to our knowledge, has not been previously reported for CHO-produced human IgG antibodies.

Treatment of post-transplantation lymphoproliferative disorders after kidney transplant with rituximab and conversion to m-TOR inhibitor / Tratamiento de la enfermedad linfoproliferativa post-trasplante renal con Rituximab y conversión a inhibidor m-TOR

Abstract Background: Post-transplantation lymphoproliferative disorders are serious complications of organ transplantation which treatment is not yet standardized. Objective: To describe the clinical response, overall and graft survival of patients in our center with this complication after kidney transplantation, which received rituximab as part of their treatment as well as conversion to m-TOR. Methods: Retrospective study, which included patients, diagnosed with post-transplant lymphoproliferative disorders after kidney transplantation from January 2011 to July 2014. Results: Eight cases were found with a wide spectrum of clinical presentations. Most had monomorphic histology, 85% were associated with Epstein-Barr virus, 25% of patients had tumor involvement of the renal graft, and 12.5% ​​had primary central nervous system lymphoma. All patients were managed with reduction of immunosuppression, conversion to m-TOR (except one who lost the graft at diagnosis) and rituximab-based therapy. The overall response rate was 87.5% (62.5% complete response, 25% partial response). Survival was 87.5% with a median follow-up of 34 months. An additional patient lost the graft, with chronic nephropathy already known. All the remaining patients had stable renal function. Conclusions: There are no standardized treatment regimens for lymphoproliferative disorders after kidney transplantation, but these patients can be managed successfully with reduction of immunosuppression, conversion to m-TOR and rituximab-based schemes.

Resumen Antecedente: La enfermedad linfoproliferativa post-trasplante es una complicación grave del trasplante de órganos cuyo tratamiento aún no se encuentra estandarizado. Objetivo: Describir la respuesta clínica, supervivencia global y del injerto en pacientes con esta complicación post trasplante renal en nuestro centro y que recibieron rituximab como parte de su tratamiento y la conversión a m-TOR. Métodos: Estudio retrospectivo que incluyó pacientes con diagnóstico de enfermedad linfoproliferativa postrasplante renal entre enero de 2011 y julio de 2014. Resultados: Se encontraron ocho casos, con presentaciones clínicas variables. La mayoría correspondieron a histología monomórfica, en 85% se asoció con virus de Epstein-Barr, 25% de los pacientes tenían compromiso tumoral del injerto renal y 12.5% linfoma primario de sistema nervioso central. Todos los pacientes se manejaron con reducción de inmunosupresión, conversión a m-TOR (excepto uno que perdió el injerto al diagnóstico) y tratamiento basado en rituximab. La tasa de respuesta global fue del 87.5% (62.5% respuesta completa, 25% respuesta parcial). La supervivencia fue del 87.5% con una mediana de seguimiento de 34 meses. Un paciente adicional perdió el injerto renal, con nefropatía crónica ya conocida. Los pacientes restantes con función renal estable. Conclusiones: No existen esquemas estandarizados de tratamiento para la enfermedad linfoproliferativa post-trasplante renal, pero estos pacientes pueden ser manejados de forma exitosa con reducción de la inmunosupresión, conversión a m-TOR y esquemas basados en rituximab.

Enfermedad linfoproliferativa postrasplante de órgano sólido / Post-transplant lymphoproliferative disease / Doença linfoproliferativa pós-transplante de órgão sólido

Con la denominación enfermedad linfoproliferativa postrasplante (ELPT) se conoce un grupo de trastornos que se pueden presentar con posterioridad al trasplante de órganos sólidos, con incidencia variable dependiendo del tipo de órgano trasplantado. La presentación clínica inespecífica de esta enfermedad, el compromiso extranodal y su amplio espectro histopatológico hacen que tanto la clasificación como el tratamiento sean complejos. Esta revisión presenta una actualización sobre la ELPT en pacientes con trasplante de órgano sólido.

Post-transplant lymphoproliferative disease (PTLD) is a group of disorders that may occur after transplantation. Its incidence is variable according to the transplanted organ. The clinical variability of this disease, its extra-nodal involvement and its broad histopathological spectrum make both classification and management very complex. This review provides an update about PTLD in patients with solid organ transplantation.

Com a denominação doença linfoproliferativa póstransplante (ELPT) se conhece um grupo de transtornos que se podem apresentar com posterioridade ao transplante de órgãos sólidos, com incidência variável dependendo do tipo de órgão transplantado. A apresentação clínica inespecífica desta doença, o compromisso extranodal e seu amplo espectro histopatológico fazem que tanto a classificação como o tratamento sejam complexos. Esta revisão apresenta uma atualização sobre a ELPT em pacientes com transplante de órgão sólido.

Treatment of post-transplantation lymphoproliferative disorders after kidney transplant with rituximab and conversion to m-TOR inhibitor.

BACKGROUND: Post-transplantation lymphoproliferative disorders are serious complications of organ transplantation which treatment is not yet standardized. OBJECTIVE: To describe the clinical response, overall and graft survival of patients in our center with this complication after kidney transplantation, which received rituximab as part of their treatment as well as conversion to m-TOR. METHODS: Retrospective study, which included patients, diagnosed with post-transplant lymphoproliferative disorders after kidney transplantation from January 2011 to July 2014. RESULTS: Eight cases were found with a wide spectrum of clinical presentations. Most had monomorphic histology, 85% were associated with Epstein-Barr virus, 25% of patients had tumor involvement of the renal graft, and 12.5% ​​had primary central nervous system lymphoma. All patients were managed with reduction of immunosuppression, conversion to m-TOR (except one who lost the graft at diagnosis) and rituximab-based therapy. The overall response rate was 87.5% (62.5% complete response, 25% partial response). Survival was 87.5% with a median follow-up of 34 months. An additional patient lost the graft, with chronic nephropathy already known. All the remaining patients had stable renal function. CONCLUSIONS: There are no standardized treatment regimens for lymphoproliferative disorders after kidney transplantation, but these patients can be managed successfully with reduction of immunosuppression, conversion to m-TOR and rituximab-based schemes.

ANTECEDENTE: La enfermedad linfoproliferativa post-trasplante es una complicación grave del trasplante de órganos cuyo tratamiento aún no se encuentra estandarizado. OBJETIVO: Describir la respuesta clínica, supervivencia global y del injerto en pacientes con esta complicación post trasplante renal en nuestro centro y que recibieron rituximab como parte de su tratamiento y la conversión a m-TOR. MÉTODOS: Estudio retrospectivo que incluyó pacientes con diagnóstico de enfermedad linfoproliferativa postrasplante renal entre enero de 2011 y julio de 2014. RESULTADOS: Se encontraron ocho casos, con presentaciones clínicas variables. La mayoría correspondieron a histología monomórfica, en 85% se asoció con virus de Epstein-Barr, 25% de los pacientes tenían compromiso tumoral del injerto renal y 12.5% linfoma primario de sistema nervioso central. Todos los pacientes se manejaron con reducción de inmunosupresión, conversión a m-TOR (excepto uno que perdió el injerto al diagnóstico) y tratamiento basado en rituximab. La tasa de respuesta global fue del 87.5% (62.5% respuesta completa, 25% respuesta parcial). La supervivencia fue del 87.5% con una mediana de seguimiento de 34 meses. Un paciente adicional perdió el injerto renal, con nefropatía crónica ya conocida. Los pacientes restantes con función renal estable. CONCLUSIONES: No existen esquemas estandarizados de tratamiento para la enfermedad linfoproliferativa post-trasplante renal, pero estos pacientes pueden ser manejados de forma exitosa con reducción de la inmunosupresión, conversión a m-TOR y esquemas basados en rituximab.

Manejo no farmacológico del dolor en pacientes en posoperatorio de revascularización miocárdica en UCI / Non-pharmacological pain management in postoperative myocardial revascularization patients in the ICU

Este es un estudio descriptivo, que tuvo como objetivo principal documentar y describir el manejo no farmacológico que le dan los enfermeros a los pacientes en PORVM en la UCI, determinar los métodos que utilizan y las características de la aplicación de las terapias no farmacológicas en este tipo de pacientes. Los principales resultados documentan que las técnicas no farmacológicas para el manejo del dolor en pacientes en posoperatorio de revascularización miocárdica, son utilizadas de forma poco frecuente y con limitaciones tales como: la falta de formación y educación en el tema, la falta de indicación terapéutica, las múltiples actividades en el horario laboral y por supuesto la falta de protocolos institucionales y guías de manejo donde se incluyan técnicas no farmacológicas, que estandaricen el manejo del dolor en el paciente en PORVM.

This is a descriptive study 's main objective was to document and describe the nonpharmacological management that give nurses to patients in the ICU PORVM determine the methods used and the characteristics of the implementation of non-pharmacological therapies in this patients. The main results documented that non -pharmacological pain management in patients with postoperative myocardial revascularization techniques are used infrequently and limitations such as lack of training and education on the subject, the lack of therapeutic indication, multiple activities in working hours and of course the lack of institutional management protocols and guidelines where no pharmacological techniques, to standardize the management of pain in patients in PORVM included.

ESR1 gene promoter region methylation in free circulating DNA and its correlation with estrogen receptor protein expression in tumor tissue in breast cancer patients.

BACKGROUND: Tumor expression of estrogen receptor (ER) is an important marker of prognosis, and is predictive of response to endocrine therapy in breast cancer. Several studies have observed that epigenetic events, such methylation of cytosines and deacetylation of histones, are involved in the complex mechanisms that regulate promoter transcription. However, the exact interplay of these factors in transcription activity is not well understood. In this study, we explored the relationship between ER expression status in tumor tissue samples and the methylation of the 5' CpG promoter region of the estrogen receptor gene (ESR1) isolated from free circulating DNA (fcDNA) in plasma samples from breast cancer patients. METHODS: Patients (n = 110) with non-metastatic breast cancer had analyses performed of ER expression (luminal phenotype in tumor tissue, by immunohistochemistry method), and the ESR1-DNA methylation status (fcDNA in plasma, by quantitative methylation specific PCR technique). RESULTS: Our results showed a significant association between presence of methylated ESR1 in patients with breast cancer and ER negative status in the tumor tissue (p = 0.0179). There was a trend towards a higher probability of ESR1-methylation in those phenotypes with poor prognosis i.e. 80% of triple negative patients, 60% of HER2 patients, compared to 28% and 5.9% of patients with better prognosis such as luminal A and luminal B, respectively. CONCLUSION: Silencing, by methylation, of the promoter region of the ESR1 affects the expression of the estrogen receptor protein in tumors of breast cancer patients; high methylation of ESR1-DNA is associated with estrogen receptor negative status which, in turn, may be implicated in the patient's resistance to hormonal treatment in breast cancer. As such, epigenetic markers in plasma may be of interest as new targets for anticancer therapy, especially with respect to endocrine treatment.

Protection provided by a herpes simplex virus 2 (HSV-2) glycoprotein C and D subunit antigen vaccine against genital HSV-2 infection in HSV-1-seropositive guinea pigs.

A prophylactic vaccine for genital herpes disease remains an elusive goal. We report the results of two studies performed collaboratively in different laboratories that assessed immunogenicity and vaccine efficacy in herpes simplex virus 1 (HSV-1)-seropositive guinea pigs immunized and subsequently challenged intravaginally with HSV-2. In study 1, HSV-2 glycoproteins C (gC2) and D (gD2) were produced in baculovirus and administered intramuscularly as monovalent or bivalent vaccines with CpG and alum. In study 2, gD2 was produced in CHO cells and given intramuscularly with monophosphoryl lipid A (MPL) and alum, or gC2 and gD2 were produced in glycoengineered Pichia pastoris and administered intramuscularly as a bivalent vaccine with Iscomatrix and alum to HSV-1-naive or -seropositive guinea pigs. In both studies, immunization boosted neutralizing antibody responses to HSV-1 and HSV-2. In study 1, immunization with gC2, gD2, or both immunogens significantly reduced the frequency of genital lesions, with the bivalent vaccine showing the greatest protection. In study 2, both vaccines were highly protective against genital disease in naive and HSV-1-seropositive animals. Comparisons between gD2 and gC2/gD2 in study 2 must be interpreted cautiously, because different adjuvants, gD2 doses, and antigen production methods were used; however, significant differences invariably favored the bivalent vaccine. Immunization of naive animals with gC2/gD2 significantly reduced the number of days of vaginal shedding of HSV-2 DNA compared with that for mock-immunized animals. Surprisingly, in both studies, immunization of HSV-1-seropositive animals had little effect on recurrent vaginal shedding of HSV-2 DNA, despite significantly reducing genital disease.

Binding of DC-SIGN to glycoproteins expressed in glycoengineered Pichia pastoris.

Previous studies have shown that glycoproteins expressed in wild-type Pichia pastoris bind to Dendritic cell-SIGN (DC-Specific Intercellular adhesion molecule-3 Grabbing Nonintegrin), a mannose-binding receptor found on dendritic cells in peripheral tissues which is involved in antigen presentation and the initiation of an immune response. However, the binding of DC-SIGN to glycoproteins purified from P. pastoris strains engineered to express humanized N- and O-linked glycans has not been tested to date. In this study, the binding of glycoproteins with specific high-mannose or human N- and O-linked glycan structures to DC-SIGN was tested. Proteins with humanized N-glycans including Man5 structures and O-glycans (up to as many as 24) with single mannose chain length showed DC-SIGN binding that was comparable to that measured for a CHO-produced IgG1 which lacks O-linked mannose. Glycoproteins with wild-type N-glycans and mannotriose and higher O-glycans bound to DC-SIGN in a manner that was strongly inhibited by either the use of enzymatic N-deglycosylation or sodium meta-periodate oxidation. Mannan purified from humanized P. pastoris also showed lower ability to inhibit DC-SIGN binding to glycoproteins with wild type fungal glycosylation than mannan purified from wild type strains. This study shows that humanized P. pastoris can produce glycoproteins that do not bind to DC-SIGN.

Conocimientos, actitudes y prácticas acerca de la medicina alternativa en médicos vinculados a hospitales públicos de Cundinamarca, Colombia / Knowledge, attitudes and practice concerning complementary and alternative medicine (CAM) amongst doctors working in public hospitals in Cundinamarca, Colombia

Objetivo: Identificar conocimientos, actitudes y practicas acerca de la medicina alternativa y complementaria en médicos vinculados a hospitales públicos en el departamento de Cundinamarca. Métodos: Se realizó un estudio cuantitativo observacional descriptivo. La población estuvo conformada por médicos generales y especialistas vinculados a los 37 hospitales públicos del departamento de Cundinamarca durante el periodo diciembre de 2009 a marzo 2010. La muestra incluyó a 280 médicos con un nivel de confianza del 95 % y error del 0,5 %. Resultados: Existe un desconocimiento para los sistemas médicos complejos del 93 % y para las terapéuticas de un 91 %. Sin embargo, el 41 % de los médicos entrevistados desea capacitarse formalmente en el área de medicina alternativa y complementaria (MAC) y el 15 % tiene alguna formación en esta área. El 18 % remiten a sus pacientes para tratamientos relacionados con MAC, el 25 % ha recibido tratamiento de MAC. La mitad de los médicos considera que se debe enseñar a los estudiantes de medicina sobre MAC. Conclusiones: Existe una tendencia positiva hacia la MAC de la experiencia y edad del médico. Además se considera que poseen una evidencia científica demostrable, por lo que desean recibir una educación formal en esta área. Existe una percepción de utilidad, uso y recomendación de estas medicinas tanto para su vida personal como en la práctica médica cotidiana, haciendo explicita la necesidad de acceso a las MAC a través del plan obligatorio de salud.

Objective: Identifying the knowledge, attitudes and practices of doctors working in public hospitals in the Cundinamarca department, Colombia, regarding complementary and alternative medicine. Methods: This was a descriptive, quantitative observational study. The target population consisted of general practitioners and specialists involved in the 37 public hospitals in the Cundinamarca department from December 2009 to March 2010. The sample included 280 doctors (using 95 % confidence level and 0.5% error). Results: There was 93 % misunderstanding of complex medical systems and 91 % regarding therapeutic systems. However, 41 % of the doctors surveyed wished to receive formal training in the area of complementary and alternative medicine (CAM) and 15 % had some training in this area. 18 % referred patients for CAM-related treatment and 25 % had received MAC treatment. Half of the doctors considered that CAM should be taught to medical students. 40 % considered that CAM should be included in the compulsory health plan in Colombia. Conclusions: A positive trend regarding CAM was related to a doctor's age and experience. It was also believed that demonstrable scientific evidence was available regarding CAM so they wished to receive formal education in this area. These drugs were perceived to be useful, in current use and recommended concerning their personal life and everyday medical practice, making explicit the need for access to MAC through the compulsory Colombian health plan.

[Knowledge, attitudes and practice concerning complementary and alternative medicine (CAM) amongst doctors working in public hospitals in Cundinamarca, Colombia]. / Conocimientos, actitudes y prácticas acerca de la medicina alternativa en médicos vinculados a hospitales públicos de Cundinamarca, Colombia.

OBJECTIVE: Identifying the knowledge, attitudes and practices of doctors working in public hospitals in the Cundinamarca department, Colombia, regarding complementary and alternative medicine. METHODS: This was a descriptive, quantitative observational study. The target population consisted of general practitioners and specialists involved in the 37 public hospitals in the Cundinamarca department from December 2009 to March 2010. The sample included 280 doctors (using 95 % confidence level and 0.5% error). RESULTS: There was 93 % misunderstanding of complex medical systems and 91 % regarding therapeutic systems. However, 41 % of the doctors surveyed wished to receive formal training in the area of complementary and alternative medicine (CAM) and 15 % had some training in this area. 18 % referred patients for CAM-related treatment and 25 % had received MAC treatment. Half of the doctors considered that CAM should be taught to medical students. 40 % considered that CAM should be included in the compulsory health plan in Colombia. CONCLUSIONS: A positive trend regarding CAM was related to a doctor's age and experience. It was also believed that demonstrable scientific evidence was available regarding CAM so they wished to receive formal education in this area. These drugs were perceived to be useful, in current use and recommended concerning their personal life and everyday medical practice, making explicit the need for access to MAC through the compulsory Colombian health plan.

Diagnosis of pulmonary and extrapulmonary tuberculosis using an in-house PCR method in clinical samples from a middle-income resource setting / Diagnóstico de tuberculosis pulmonar y extrapulmonar mediante un método de PCR casera en muestras clínicas de una región de desarrollo intermedio

Introduction: PCR detection offers a good opportunity to obtain fast results which is a priority in tuberculosis control programs. Objectives: We assayed an in-house PCR method based on the detection of mycobaterial IS6110 gene in clinical samples of patients with pulmonary and extrapulmonary tuberculosis to demonstrate its usefulness and reliability in the setting of a middle-resource region with high tuberculosis prevalence. Materials and methods: Pulmonary (n=317) and extrapulmonary (n=41) samples were collected from 358 patients with clinical suspicion of tuberculosis. All samples were processed to detect acid-fast bacilli by microscopy, culture on solid media and PCR. To remove PCR inhibitors, three washing steps of the decontaminated pellet were included before mycobacterial cell lysis. Results: The overall sensitivity was 96% in clinical samples, and specificity was 100% for our in-house method in pulmonary and extrapulmonary samples. No inhibition was found among samples that were PCR negative, but culture positive for Mycobacterium tuberculosis. No false positives were found. Conclusions: In-house PCR in a middle-income setting region, with simple and strictly controlled methods, could efficiently complement conventional bacteriological tools for the rapid diagnosis of tuberculosis, especially in paucibacillary and extrapulmonary samples.

Introducción. La prueba de PCR ofrece la posibilidad de obtener resultados rápidos, una prioridad para los programas de control de la tuberculosis. Objetivos.Evaluar el método de PCR basado en la detección del gen IS6110 de las micobacterias en muestras clínicas de pacientes con tuberculosis pulmonar y extrapulmonar, para demostrar su utilidad y confiabilidad en un sitio de desarrollo medio con alta prevalencia de tuberculosis. Materiales y métodos. Se recolectaron 317 muestras pulmonares y 41 de origen extrapulmonar, de 358 pacientes con sospecha clínica de tuberculosis. Todas las muestras se procesaron para detectar bacilos ácido-alcohol resistentes por microscopía, cultivo en medio sólido y PCR. Para eliminar los inhibidores de la PCR se lavaron tres veces antes de realizar la lisis de la pared de la micobacteria. Resultados. La sensibilidad total fue de 96% y la especificidad de 100% para este método "casero" en muestras pulmonares y extrapulmonares. No se encontró inhibición entre las muestras que fueron negativas por PCR pero con cultivo positivo para Mycobacterium tuberculosis. No se hallaron falsos positivos. Conclusiones. La implementación de la PCR "casera" en regiones de desarrollo intermedio como método simple y estrictamente controlado, podría complementar eficientemente las herramientas bacteriológicas tradicionales para el diagnóstico rápido de la tuberculosis, especialmente en muestras paucibacilares y de origen extrapulmonar.