Establishment and Validation of Pathogenic CS17+ and CS19+ Enterotoxigenic Escherichia coli Challenge Models in the New World Primate Aotus nancymaae.

Enterotoxigenic Escherichia coli (ETEC) is a common cause of diarrheal illness in the military, travelers, and children living in low- to middle-income countries. Increased antibiotic resistance, the absence of a licensed vaccine, and the lack of broadly practical therapeutics perpetuate the significant health and financial burden resulting from ETEC infection. A critical step in the evaluation of vaccines and therapeutics is preclinical screening in a relevant animal disease model that closely replicates human disease. We previously developed a diarrheal model of class 5a colonization factor (CF) CFA/I-expressing ETEC in the New World owl monkey species Aotus nancymaae using ETEC strain H10407. In order to broaden the use of the model, we report here on the development of A. nancymaae models of ETEC expressing the class 5b CFs CS17 and CS19 with strains LSN03-016011/A and WS0115A, respectively. For both models, we observed diarrheal attack rates of ≥80% after oral inoculation with 5 × 1011 CFU of bacteria. These models will aid in assessing the efficacy of future ETEC vaccine candidates and therapeutics.

Establishment, Validation, and Application of a New World Primate Model of Enterotoxigenic Escherichia coli Disease for Vaccine Development.

The establishment of an animal model that closely approximates enterotoxigenic Escherichia coli (ETEC) disease in humans is critical for the development and evaluation of vaccines against this enteropathogen. Here, we evaluated the susceptibility of Aotus nancymaae, a New World monkey species, to ETEC infection. Animals were challenged orogastrically with 109 to 1011 CFU of the human pathogenic CFA/I+ ETEC strain H10407 and examined for evidence of diarrhea and fecal shedding of bacteria. A clear dose-range effect was obtained, with diarrheal attack rates of 40% to 80%, validated in a follow-on study demonstrating an attack rate of 80% with 1011 CFU of H10407 ETEC. To determine whether this model is an effective approach for assessing ETEC vaccine candidates, we used it to evaluate the ability of the donor strand-complemented CFA/I adhesin CfaE (dscCfaE) to protect against H10407 challenge. In a series of experiments, animals were intranasally vaccinated with dscCfaE alone, dscCfaE with either cholera toxin B-subunit (CTB) or heat-labile toxin (LTB), or phosphate-buffered saline (PBS) alone and then challenged with 1011 CFU of H10407. Control animals vaccinated with PBS had attack rates of 70 to 90% on challenge. Vaccination with dscCfaE, or dscCfaE admixed with CTB or LTB, resulted in a reduction of attack rates, with vaccine efficacies of 66.7% (P = 0.02), 77.7% (P = 0.006), and 42.9% (P = 0.370) to 83.3% (P = 0.041), respectively. In conclusion, we have shown the H10407 ETEC challenge of A. nancymaae to be an effective, reproducible model of ETEC disease, and importantly, we have demonstrated that in this model, vaccination with the prototype vaccine candidate dscCfaE is protective against CF-homologous disease.

Antimicrobial susceptibility of Brucella melitensis isolates in Peru.

Brucellosis is an important public health problem in Peru. We evaluated 48 human Brucella melitensis biotype 1 strains from Peru between 2000 and 2006. MICs of isolates to doxycycline, azithromycin, gentamicin, rifampin, ciprofloxacin, and trimethoprim-sulfamethoxazole were determined by the Etest method. All isolates were sensitive to tested drugs during the periods of testing. Relapses did not appear to be related to drug resistance.

Enhanced Immunogenicity and Protective Efficacy of a Campylobacter jejuni Conjugate Vaccine Coadministered with Liposomes Containing Monophosphoryl Lipid A and QS-21.

Campylobacter jejuni is among the most common causes of diarrheal disease worldwide and efforts to develop protective measures against the pathogen are ongoing. One of the few defined virulence factors targeted for vaccine development is the capsule polysaccharide (CPS). We have developed a capsule conjugate vaccine against C. jejuni strain 81-176 (CPS-CRM) that is immunogenic in mice and nonhuman primates (NHPs) but only moderately immunogenic in humans when delivered alone or with aluminum hydroxide. To enhance immunogenicity, two novel liposome-based adjuvant systems, the Army Liposome Formulation (ALF), containing synthetic monophosphoryl lipid A, and ALF plus QS-21 (ALFQ), were evaluated with CPS-CRM in this study. In mice, ALF and ALFQ induced similar amounts of CPS-specific IgG that was significantly higher than levels induced by CPS-CRM alone. Qualitative differences in antibody responses were observed where CPS-CRM alone induced Th2-biased IgG1, whereas ALF and ALFQ enhanced Th1-mediated anti-CPS IgG2b and IgG2c and generated functional bactericidal antibody titers. CPS-CRM + ALFQ was superior to vaccine alone or CPS-CRM + ALF in augmenting antigen-specific Th1, Th2, and Th17 cytokine responses and a significantly higher proportion of CD4+ IFN-γ+ IL-2+ TNF-α+ and CD4+ IL-4+ IL-10+ T cells. ALFQ also significantly enhanced anti-CPS responses in NHPs when delivered with CPS-CRM compared to alum- or ALF-adjuvanted groups and showed the highest protective efficacy against diarrhea following orogastric challenge with C. jejuni This study provides evidence that the ALF adjuvants may provide enhanced immunogenicity of this and other novel C. jejuni capsule conjugate vaccines in humans.IMPORTANCECampylobacter jejuni is a leading cause of diarrheal disease worldwide, and currently no preventative interventions are available. C. jejuni is an invasive mucosal pathogen that has a variety of polysaccharide structures on its surface, including a capsule. In phase 1 studies, a C. jejuni capsule conjugate vaccine was safe but poorly immunogenic when delivered alone or with aluminum hydroxide. Here, we report enhanced immunogenicity of the conjugate vaccine delivered with liposome adjuvants containing monophosphoryl lipid A without or with QS-21, known as ALF and ALFQ, respectively, in preclinical studies. Both liposome adjuvants significantly enhanced immunity in mice and nonhuman primates and improved protective efficacy of the vaccine compared to alum in a nonhuman primate C. jejuni diarrhea model, providing promising evidence that these potent adjuvant formulations may enhance immunogenicity in upcoming human studies with this C. jejuni conjugate and other malaria and HIV vaccine platforms.

[Preliminary results of the strengthening of the national death registry information system]. / Resultados preliminares del fortalecimiento del sistema informático nacional de defunciones.

Peru has a low coverage of deaths with a cause of death (54%) and a poor-quality registration of causes of death, as about 30% of causes of death are classified as poorly-defined or not very useful for the formulation of public policies. In response to these problems, the Ministry of Health, together with other government agencies, with the support of the Bloomberg Philanthropies "Data for Health Initiative," is implementing the National Death Registry Information System (SINADEF). The objective of this article is to describe the process of strengthening the mortality information system in Peru, focused on the implementation of SINADEF. The activities that have been carried out are described in the following areas: a) Management of the mortality information system, b) Process standardization, c) Use of information and communication technology, d) Coverage of deaths with medical certificate, e) Improvement of the quality of information, f) Development of studies, and g) Monitoring of processes. Since the implementation of SINADEF in August 2016 until July 2018, 28,407 users of the SINADEF application have been created and a total of 122,411 deaths have been registered. The quality of data recording, including the cause of death, has been improved, while low coverage of deaths with a cause of death still persists.

El Perú tiene una baja cobertura de defunciones con causa de defunción (54 %) y una mala calidad del registro de las causas de defunción, mas de 45 % de las causas de muerte se clasifican como mal definidas o poco útiles para la formulación de políticas públicas. En respuesta a estos problemas, el Ministerio de Salud, junto a otras agencias gubernamentales, con el apoyo de la Iniciativa Bloomberg "Información para la Salud" está implementando el Sistema Informático Nacional de Defunciones (SINADEF). El objetivo de este artículo es describir el proceso de fortalecimiento del sistema de información de la mortalidad en Perú, centrado en la implementación del SINADEF. Se describe las actividades que se vienen realizando en los siguientes ejes: a) Gestión del sistema de información de la mortalidad, b) Estandarización de procesos, c) Uso de tecnología de información y comunicación, d) Cobertura de las defunciones con certificación médica, e) Mejora de la calidad de la información, f) Desarrollo de estudios y g) Monitoreo de los procesos. Desde el inicio de la implementación del SINADEF, en agosto de 2016 hasta julio de 2018, se han creado 28 407 usuarios del aplicativo del SINADEF y se han registrado un total de 122 411 defunciones. Se ha mejorado la calidad del registro de los datos, incluyendo la causa de defunción, pero aún persiste la baja cobertura de defunciones con causa de muerte.

Immunogenicity of a prototype enterotoxigenic Escherichia coli adhesin vaccine in mice and nonhuman primates.

Enterotoxigenic Escherichia coli (ETEC) are the most common cause of bacterial diarrhea in young children in developing countries and in travelers. Efforts to develop an ETEC vaccine have intensified in the past decade, and intestinal colonization factors (CFs) are somatic components of most investigational vaccines. CFA/I and related Class 5 fimbrial CFs feature a major stalk-forming subunit and a minor, antigenically conserved tip adhesin. We hypothesized that the tip adhesin is critical for stimulating antibodies that specifically inhibit ETEC attachment to the small intestine. To address this, we compared the capacity of donor strand complemented CfaE (dscCfaE), a stabilized form of the CFA/I fimbrial tip adhesin, and CFA/I fimbriae to elicit anti-adhesive antibodies in mice, using hemagglutination inhibition (HAI) as proxy for neutralization of intestinal adhesion. When given with genetically attenuated heat-labile enterotoxin LTR192G as adjuvant by intranasal (IN) or orogastric (OG) vaccination, dscCfaE exceeded CFA/I fimbriae in eliciting serum HAI titers and anti-CfaE antibody titers. Based on these findings, we vaccinated Aotus nancymaae nonhuman primates (NHP) with dscCfaE alone or admixed with one of two adjuvants, LTR192G and cholera toxin B-subunit, by IN and OG administration. Only IN vaccination with dscCfaE with either adjuvant elicited substantial serum HAI titers and IgA and IgG anti-adhesin responses, with the latter detectable a year after vaccination. In conclusion, we have shown that dscCfaE elicits robust HAI and anti-adhesin antibody responses in both mice and NHPs when given with adjuvant by IN vaccination, encouraging further evaluation of an ETEC adhesin-based vaccine approach.

[Information system of the national network of public health laboratories in Peru (Netlab)]. / Sistema de información de la red nacional de laboratorios de salud pública en el Perú (Netlab).

Clinical laboratory information systems produce improvements in the quality of information, reduce service costs, and diminish wait times for results, among other things. In the construction process of this information system, the National Institute of Health (NIH) of Peru has developed and implemented a web-based application to communicate to health personnel (laboratory workers, epidemiologists, health strategy managers, physicians, etc.) the results of laboratory tests performed at the Peruvian NIH or in the laboratories of the National Network of Public Health Laboratories which is called NETLAB. This article presents the experience of implementing NETLAB, its current situation, perspectives of its use, and its contribution to the prevention and control of diseases in Peru.

Resultados preliminares del fortalecimiento del sistema informático nacional de defunciones / Preliminary results of the strengthening of the national death registry information system

El Perú tiene una baja cobertura de defunciones con causa de defunción (54 %) y una mala calidad del registro de las causas de defunción, mas de 45 % de las causas de muerte se clasifican como mal definidas o poco útiles para la formulación de políticas públicas. En respuesta a estos problemas, el Ministerio de Salud, junto a otras agencias gubernamentales, con el apoyo de la Iniciativa Bloomberg «Información para la Salud¼ está implementando el Sistema Informático Nacional de Defunciones (SINADEF). El objetivo de este artículo es describir el proceso de fortalecimiento del sistema de información de la mortalidad en Perú, centrado en la implementación del SINADEF. Se describe las actividades que se vienen realizando en los siguientes ejes: a) Gestión del sistema de información de la mortalidad, b) Estandarización de procesos, c) Uso de tecnología de información y comunicación, d) Cobertura de las defunciones con certificación médica, e) Mejora de la calidad de la información, f) Desarrollo de estudios y g) Monitoreo de los procesos. Desde el inicio de la implementación del SINADEF, en agosto de 2016 hasta julio de 2018, se han creado 28 407 usuarios del aplicativo del SINADEF y se han registrado un total de 122 411 defunciones. Se ha mejorado la calidad del registro de los datos, incluyendo la causa de defunción, pero aún persiste la baja cobertura de defunciones con causa de muerte.

Peru has a low coverage of deaths with a cause of death (54%) and a poor-quality registration of causes of death, as about 30% of causes of death are classified as poorly-defined or not very useful for the formulation of public policies. In response to these problems, the Ministry of Health, together with other government agencies, with the support of the Bloomberg Philanthropies «Data for Health Initiative,¼ is implementing the National Death Registry Information System (SINADEF). The objective of this article is to describe the process of strengthening the mortality information system in Peru, focused on the implementation of SINADEF. The activities that have been carried out are described in the following areas: a) Management of the mortality information system, b) Process standardization, c) Use of information and communication technology, d) Coverage of deaths with medical certificate, e) Improvement of the quality of information, f) Development of studies, and g) Monitoring of processes. Since the implementation of SINADEF in August 2016 until July 2018, 28,407 users of the SINADEF application have been created and a total of 122,411 deaths have been registered. The quality of data recording, including the cause of death, has been improved, while low coverage of deaths with a cause of death still persists.

Sistema de información de la red nacional de laboratorios de salud pública en el Perú (Netlab) / Information system of the national network of public health laboratories in Peru (Netlab)

Los sistemas de información de laboratorio clínico, producen mejoras en la calidad de la información, la reducción de los costos del servicio, disminución de la espera para obtener resultados, entre otros. En el proceso de construcción de este sistema de información, el Instituto Nacional de Salud (INS) del Perú ha desarrollado e implementado un aplicativo a través de Internet basado en la web, para comunicar, al personal de salud (laboratoristas, epidemiólogos, gestores de estrategias sanitarias, médicos tratantes, etc.), los resultados de las pruebas de laboratorio que se realizan en el INS o en los laboratorios de la Red Nacional de Laboratorios de Salud Pública el cual es llamado NETLAB. Este artículo presenta la experiencia de la implementación de NETLAB, su situación actual y las perspectivas de su empleo, así como su contribución en la prevención y control de enfermedades en el Perú.

Clinical laboratory information systems produce improvements in the quality of information, reduce service costs, and diminish wait times for results, among other things. In the construction process of this information system, the National Institute of Health (NIH) of Peru has developed and implemented a web-based application to communicate to health personnel (laboratory workers, epidemiologists, health strategy managers, physicians, etc.) the results of laboratory tests performed at the Peruvian NIH or in the laboratories of the National Network of Public Health Laboratories which is called NETLAB. This article presents the experience of implementing NETLAB, its current situation, perspectives of its use, and its contribution to the prevention and control of diseases in Perú.

The effect of preparation of cebiche on the survival of enterotoxigenic Escherichia coli, Aeromonas hydrophila, and Vibrio parahaemolyticus.

BACKGROUND: Cebiche is a common dish in Latin America, prepared using raw fish mixed with vegetables and marinated with lime juice. The acidity of the lime juice is commonly believed to destroy bacteria and render cebiche as safe to eat. Little data exist concerning rates of cebiche-associated gastroenteritis outbreaks, although these may be high given the popularity of the dish. METHODS: We inoculated raw fish with Aeromonas hydrophila, Vibrio parahaemolyticus, and enterotoxigenic Escherichia coli to determine the effect of the cebiche preparation process on bacterial viability. Raw fish were exposed to a suspension of 1.0 × 10(8) colony-forming units (CFUs) of each organism in a 50-mL solution, prior to the addition of cebiche ingredients. A typical Peruvian cebiche recipe was used combining limes, onions, sweet potatoes, cilantro, and hot peppers marinated together for 30 minutes. A homogenized mixture of the dish was then evaluated for pH and bacterial counts at 0, 10, and 30 minutes. As much as 100 µL of inocula were streaked onto tryptic soy agar (TSA) agar plates and incubated for 24 hours. RESULTS: The initial average pH of the fish was 6.4 prior to adding cebiche ingredients and 5.0 immediately afterwards. The pH at 10- and 30-minute periods was 5.4 and 5.2, respectively. Little reduction in bacterial counts was observed at either the 10- or 30-minute time periods, with counts increasing at 30 minutes. CONCLUSIONS: The putative bactericidal role of lime juice in the preparation process is not sufficient to reduce the microbial population present in cebiche. Pathogens may remain viable after exposure to acidic conditions. The increasing popularity of Peruvian cuisine may also lead to cebiche-associated illness outside of Latin America.

New World monkey Aotus nancymae as a model for Campylobacter jejuni infection and immunity.

Three groups of six monkeys (Aotus nancymae) each were inoculated intragastrically with increasing doses of Campylobacter jejuni. Infection resulted in fecal colonization (100% of monkeys), dose-related diarrhea, and robust immune responses. Colonization duration and diarrhea rate were reduced upon secondary challenge. A. nancymae may be useful for studying anti-Campylobacter vaccine efficacy.

The New World primate, Aotus nancymae, as a model for examining the immunogenicity of a prototype enterotoxigenic Escherichia coli subunit vaccine.

The colonization factors (CF) of enterotoxigenic Escherichia coli (ETEC) are being targeted for inclusion in a multi-subunit ETEC vaccine. This study was designed to examine the preclinical safety and immunogenicity of CF CS6, encapsulated in a biodegradable poly(DL-lactide-co-glycolide) (meCS6), and administered in the presence or absence of a mutated heat-labile enterotoxin, LT(R192G), in the non-human primate, Aotus nancymae. A. nancymae were inoculated intranasally (IN) with meCS6 (200 microg; positive control), or intragastrically (IG) with meCS6 (200 or 1000 microg) with or without 2 microg LT(R192G) in three doses given at 2-week intervals. In a second experiment, A. nancymae were inoculated IG with 950 microg of meCS6 with or without 2 microg LT(R192G) in four doses given every 48 h. Blood was collected to assess anti-CS6 and -LT serum immunoglobulin G (IgG) and IgA responses and safety variables (complete blood count and chemistry). Safety parameters were unchanged from baseline following all vaccinations. In Experiment 1, a dose-related serologic response to CS6 was observed; 78.6 and 57.1% of monkeys given 1000 microg meCS6 (n = 14) had a serum IgG and IgA response, respectively, compared to only 28.6% of monkeys given 200 microg meCS6 (n = 14) with a serum IgG and IgA response. No significant effect on the number of responders or the magnitude of responses was observed with the addition of LT(R192G). The three-dose, 2-week regimen with 1000 microg meCS6 was more effective at eliciting an immune response than the four-dose, 48-h regimen with 950 microg meCS6. Results from this study indicate that A. nancymae provide a useful ETEC preclinical safety and immunogenicity model.

N-Aryl-gamma-lactams as integrin alphavbeta3 antagonists.

Novel alphavbeta3 antagonists based on the N-aryl-gamma-lactam scaffold were prepared. SAR studies led to the identification of potent antagonists for alphavbeta3 receptor with excellent selectivity against the structurally related alpha(IIb)beta3 receptor. Additional interactions of N-aryl-gamma-lactam derivatives with alphavbeta3 were found when compared to c(-RGDf[NMe]V-) peptide antagonist. The effects of the conformation and configuration of the gamma-lactam core on the binding were also assessed.

Modelo de organización y funciones de la Oficina/Unidad Regional de Investigación en Salud / Model of organization and functions of the Office / Regional Health Research Unit

El documento técnico, plasma una propuesta de organización, funciones y requisitos, que deberían cumplir las oficinas o unidades de investigación; con el objetivo de promover la investigación en salud para generar evidencias que permitan una mejor toma de decisiones, teniendo como base la descentralización defunciones y las prioridades de investigación en salud a nivel regional y nacional; así como también el respeto de las normas éticas, las buenas prácticas clínicas, normas nacionales relacionadas y los estándares dentro de las prácticas de investigación en salud