A Diversity Covering (DiCo) Plasmodium vivax apical membrane antigen-1 vaccine adjuvanted with RFASE/RSL10 yields high levels of growth-inhibitory antibodies.

Plasmodium vivax malaria is increasingly recognized as a major global health problem and the socio-economic impact of P.vivax-induced burden is huge. Vaccine development against P. vivax malaria has been hampered by the lack of an in vitro culture system and poor access to P. vivax sporozoites. The recent generation of Plasmodium falciparum parasites that express a functional P. vivax AMA1 molecule has provided a platform for in vitro evaluation of PvAMA1 as a potential blood stage vaccine. Three so-called PvAMA1 Diversity Covering (DiCo) proteins were designed to assess their potential to induce a functional and broad humoral immune response to the polymorphic PvAMA1 molecule. Rabbits were immunized with the mixture of three, Pichia-produced, PvAMA1 DiCo proteins, as well as with 2 naturally occurring PvAMA1 alleles. For these three groups, the experimental adjuvant raffinose fatty acid sulfate ester (RFASE) was used, while in a fourth group the purified main mono-esterified constituent (RSL10) of this adjuvant was used. Animals immunized with the mixture of the three PvAMA1 DiCo proteins in RFASE showed high anti-PvAMA1 antibody titers against three naturally occurring PvAMA1variants while also high growth-inhibitory capacity was observed against P. falciparum parasites expressing PvAMA1. This supports further clinical development of the PvAMA1 DiCo mixture as a potential malaria vaccine. However, as the single allele PvAMA1 SalI-group showed similar characteristics in antibody titer and inhibition levels as the PvAMA1 DiCo mixture-group, this raises the question whether a mixture is really necessary to overcome the polymorphism in the vaccine candidate. RFASE induced strong humoral responses, as did the animals immunized with the purified component, RSL10. This suggests that RSL10 is the active ingredient. However, one of the RSL10-immunized animal showed a delayed response, necessitating further research into the clinical development of RSL10.

Accelerated phase Ia/b evaluation of the malaria vaccine candidate PfAMA1 DiCo demonstrates broadening of humoral immune responses.

Plasmodium falciparum apical membrane antigen 1 (PfAMA1) is a candidate malaria vaccine antigen expressed on merozoites and sporozoites. PfAMA1's polymorphic nature impacts vaccine-induced protection. To address polymorphism, three Diversity Covering (DiCo) protein sequences were designed and tested in a staggered phase Ia/b trial. A cohort of malaria-naive adults received PfAMA1-DiCo adjuvanted with Alhydrogel® or GLA-SE and a cohort of malaria-exposed adults received placebo or GLA-SE adjuvanted PfAMA1 DiCo at weeks 0, 4 and 26. IgG and GIA levels measured 4 weeks after the third vaccination are similar in malaria-naive volunteers and placebo-immunised malaria-exposed adults, and have a similar breadth. Vaccination of malaria-exposed adults results in significant antibody level increases to the DiCo variants, but not to naturally occurring PfAMA1 variants. Moreover, GIA levels do not increase following vaccination. Future research will need to focus on stronger adjuvants and/or adapted vaccination regimens, to induce potentially protective responses in the target group of the vaccine.

A Bacterially-Expressed Recombinant Envelope Protein from Usutu Virus Induces Neutralizing Antibodies in Rabbits.

BACKGROUND: Recently, an emerging flavivirus, Usutu virus (USUV), has caused an epidemic among birds in Europe, resulting in a massive die-off in Eurasian blackbirds. Currently found only in Europe and Africa, it can be envisioned that Usutu virus will follow the path of other flaviviruses, like West Nile virus and Zika virus, and will spread via its mosquito vectors and bird hosts to other parts of the world. Several cases of human infections by Usutu virus have already been published. Anticipating this spread, development of an efficacious vaccine would be highly desirable. METHOD: This study describes the production in E. coli, purification, and refolding of a partial USUV envelope protein. Prior to immunization, the protein was characterized using size exclusion chromatography, transmission electron microscopy and dynamic light scattering, showing the limited presence of virus-like structures, indicating that the protein solution is probably a mixture of mono and multimeric envelope proteins. RESULTS: Immunizations of two rabbits with the refolded E-protein fraction, mixed with a strong adjuvant, resulted in the generation of neutralizing antibodies, as evidenced in an in vitro assay. DISCUSSION: The way forward towards a subunit vaccine against Usutu virus infection is discussed.

Evaluation of Chimpanzee Adenovirus and MVA Expressing TRAP and CSP from Plasmodium cynomolgi to Prevent Malaria Relapse in Nonhuman Primates.

Plasmodium vivax is the world's most widely distributed human malaria parasite, with over 2.8 billion people at risk in Asia, the Americas, and Africa. The 80-90% new P. vivax malaria infections are due to relapses which suggest that a vaccine with high efficacy against relapses by prevention of hypnozoite formation could lead to a significant reduction in the prevalence of P. vivax infections. Here, we describe the development of new recombinant ChAdOx1 and MVA vectors expressing P. cynomolgi Thrombospondin Related Adhesive Protein (PcTRAP) and the circumsporozoite protein (PcCSP). Both were shown to be immunogenic in mice prior to their assessment in rhesus macaques. We confirmed good vaccine-induced humoral and cellular responses after prime-boost vaccination in rhesus macaques prior to sporozoite challenge. Results indicate that there were no significant differences between mock-control and vaccinated animals after challenge, in terms of protective efficacy measured as the time taken to 1st patency, or as number of relapses. This suggests that under the conditions tested, the vaccination with PcTRAP and PcCSP using ChAdOx1 or MVA vaccine platforms do not protect against pre-erythrocytic malaria or relapses despite good immunogenicity induced by the viral-vectored vaccines.

Agenesia congénita de clítoris y labios menores en una niña de 8 años / Congenital agensis of clitoris and labia minora in 8-year-old girl

Resumen Antecedentes: La agenesia congénita del clítoris y los labios menores es una malformación poco común y los casos clínicos reportados en la bibliografía son escasos. Caso clínico: Paciente de ocho años llevada al pediatra por probable infección de vías urinarias por leucocituria. Le prescribieron un antibiótico y debido a que persistió la leucocituria le revisaron los genitales externos que se encontraron con enrojecimiento de los labios mayores y del vestíbulo vaginal, con leucorrea, sin clítoris ni labios menores. Conclusiones: Se trata de un caso de agenesia congénita de clítoris y labios menores que se suma a los escasos reportes de esta malformación.

Abstract Background: Congenital agenesis of clitoris and labia minora is a rare malformation, and clinical cases reported in the medical literature are scares. Clinical case: Female from 8 years of age, with suspicion of urinary tract infection by leucocyturia, before the failure of treatment with antibiotic and persistence of the leukocyturia, the external genital review, finding redness of labia majora and the vaginal vestibule and leukorrhea, but there no clitoris or labia minora. Conclusions: This clinical case of congenital and labia minora, joins the few scientific reports of malformation. Embryological review of how this anomaly is carried out is done.

[About the thesis of specialization in Medicine]. / A propósito de las tesis de especialidad en medicina.

No Abstract.

Sin abstract.

Low levels of polymorphisms and no evidence for diversifying selection on the Plasmodium knowlesi Apical Membrane Antigen 1 gene.

Infection with Plasmodium knowlesi, a zoonotic primate malaria, is a growing human health problem in Southeast Asia. P. knowlesi is being used in malaria vaccine studies, and a number of proteins are being considered as candidate malaria vaccine antigens, including the Apical Membrane Antigen 1 (AMA1). In order to determine genetic diversity of the ama1 gene and to identify epitopes of AMA1 under strongest immune selection, the ama1 gene of 52 P. knowlesi isolates derived from human infections was sequenced. Sequence analysis of isolates from two geographically isolated regions in Sarawak showed that polymorphism in the protein is low compared to that of AMA1 of the major human malaria parasites, P. falciparum and P. vivax. Although the number of haplotypes was 27, the frequency of mutations at the majority of the polymorphic positions was low, and only six positions had a variance frequency higher than 10%. Only two positions had more than one alternative amino acid. Interestingly, three of the high-frequency polymorphic sites correspond to invariant sites in PfAMA1 or PvAMA1. Statistically significant differences in the quantity of three of the six high frequency mutations were observed between the two regions. These analyses suggest that the pkama1 gene is not under balancing selection, as observed for pfama1 and pvama1, and that the PkAMA1 protein is not a primary target for protective humoral immune responses in their reservoir macaque hosts, unlike PfAMA1 and PvAMA1 in humans. The low level of polymorphism justifies the development of a single allele PkAMA1-based vaccine.

Diversity covering AMA1-MSP119 fusion proteins as malaria vaccines.

To overcome polymorphism in the malaria vaccine candidate Plasmodium falciparum apical membrane antigen 1 (PfAMA1), fusion protein chimeras comprised of three diversity-covering (DiCo) PfAMA1 molecules (D1, D2, and D3) and two allelic variants of the C-terminal 19-kDa region of merozoite surface protein 1 (MSP119) (variants M1 and M2) were generated. A mixture of fusion proteins (D1M1/D2M2D3) and the D1M1D2M2D3 fusion were compared to a single-unit mixture (D1/D2/D3/M1) in an immunological study in groups of rabbits. Following immunization, titers of antibodies (Abs) against four naturally occurring PfAMA1 alleles were high for all groups, as were growth inhibition assay (GIA) levels against two antigenically distinct laboratory parasite strains. Fusion of AMA1 to MSP119 did not suppress levels of antibodies against the AMA1 component. In addition, the breadth of antibody responses was unaffected. Anti-AMA1 antibodies were largely responsible for parasite growth inhibition, as shown in reversal-of-inhibition experiments by adding competing AMA1 antigen. For all groups, titration of the MSP119 antigen into the GIA led to only a small decrease in parasite inhibition, although titers of antibodies against MSP119 were increased 15-fold for the groups immunized with fusion proteins. GIA with affinity-purified anti-MSP119 antibodies showed that the 50% inhibitory concentrations of the anti-MSP119 antibody preparations were in the same order of magnitude for all animals tested, leading to the conclusion that fusing MSP119 to PfAMA1 leads to a small but significant increase in functional antibody levels. This study shows that combination of multiple vaccine candidates in fusion proteins may lead to improved characteristics of the vaccine.

Anemia del embarazo en mujeres que viven a nivel del mar / Pregnancy anemia in women living at sea level

La anemia del embarazo sigue siendo un problema de salud pública, razón por la cual en el presente trabajo se determinó la prevalencia de anemia, niveles de hemoglobina y su comportamiento por trimestre durante el embarazo en mujeres que viven a nivel del mar mediante la realización de un estudio descriptivo, transversal y prospectivo, en mujeres embarazadas de Minatitlán, Veracruz. Se estudiaron 321 mujeres embarazadas. El promedio de hemoglobina fue de 10,8 ± 0,9 g/dl. La prevalencia de anemia se presentó en 60 (18,7%) tomando como valor de referencia una hemoglobina ≤ a 10 g/dl. Las mujeres embarazadas que se encontraban en el primer trimestre presentaron en promedio una mayor cantidad de hemoglobina comparada con los dos últimos trimestres del embarazo (p<0,001). La media de hemoglobina fue de 11,5 ± 0,8 g/dl en el primer trimestre, de 10,7 ± 0,8 g/dl en el segundo trimestre y de 10,6 ± 1 g/dl en el tercer trimestre.

The main objective of this study was to determine, in pregnant women living at sea level, the prevalence of anemia (hemoglobin <10 g/dL) and hemoglobin levels duringdifferenttrimesters of pregnancy, for which it was designed an observational, cross-sectional analytical study on a secondary hospital in Minatitlan Veracruz. Were been studied 321 women, who had mean of hemoglobin 10,8 ± 0,9 g / dL, the prevalence of anemia was 18,7%. Women who were in the first trimester of pregnancy had high level of hemoglobin 11,5 ± 0,8 g / dl, which was statistically significant (p <0,001) when they were compared with the mean hemoglobin in the second (10,7 ± 0,8 g / dL) and third trimester (10,6 ± 1,0g/dL).

[Oral signs in patients with type-2 diabetes, hypertension and obesity]. / Manifestaciones bucales en pacientes con diabetes mellitus tipo 2, hipertensión y obesidad.

OBJECTIVE: To evaluate oral health in patients with type-2 diabetes, hypertension and obesity. MATERIAL AND METHODS: Comparative cross-sectional study with a random conglomerate sample, constituted by 363 patients of primary care units in Guadalajara, with records of glycemia, blood pressure and body mass index. A questionnaire was applied to know sociodemographic data, as well as medical and oral state. It included CPO-D and functionality index. For the intergroup comparison, the chi2 and t of Student were applied, considering significant p < 0.05. RESULTS: In the sample, the average age was 50 years old, 74% were women, 75 % showed overweight-obesity, arterial hypertension or diabetes mellitus; the rest had different combinations of these pathologies. In diabetic patients, glucose was of 184 +/- 74.9 mg/dL; in hypertensive ones, blood pressure was of 132/83 mm Hg, and in patients with overweight-obesity, the BMI was 34 +/- 5.33. The CPO and the functionality indexes were inadequate in all the patients. The hypertensive group showed the most deteriorated oral state; the periodontal disease prevailed. CONCLUSIONS: We propose to emphasize clinical-preventive programs in stomatologic attention of chronic patients, and to consider them a high priority. We also suggest to train the health staff and patients in the interaction between systemic illness and oral state. Finally, we propose to integrate stomatology to the research of chronic degenerative pathologies.

[Prevalence of low weight at birth and related factors]. / Prevalencia de bajo peso al nacer y factores asociados.

OBJECTIVE: To measure the prevalence of low birth weight and its possible association with risk characteristics at an international level. PATIENTS AND METHODS: An analytical and transversal study was performed during 2003. Newborns attended at the gynecological unit of the Hospital General de Zona Num. 35 in Cosamaloapan, Veracruz, were selected. The neonates included had a neonatal screening for hypothyroidism, as well as a newborn registration format. Pediatricians and pre-graduate interns evaluated: weight at birth, sex, maternal age, weeks of gestation, number of pregnancies, place of residence and the mother's occupation. Low birth weight was defined as the newborns that weighed less than 2,500 g. RESULTS: A total of 751 newborns were evaluated. In accordance to the characteristics of the mother, the mean age was of 25.9 +/- 5.9 years, 104 (13.8%) were adolescents, and 256 (35.3%) were first time mothers. The low birth weight existed in 43 (5.7%) of the newborns, and the birth weight of 708 (94.3%) was > or = 2,500 g. The maternal age, the number of pregnancies, the place of residence and the mother's occupation were not statistically related with the low birth weight. CONCLUSIONS: The prevalence of low birth weight is lower than the data reported internationally, and it could be explained by the period in which the study was performed. None of the characteristics assessed as risk factors demonstrated to be associated.

Prevalencia de asma ocupacional en trabajadores expuestos pertenecientes al municipio Guantánamo / Occupational asthama prevalence in exposed workers belonging to Guantánamo Municipality

Se realizó un estudio de corte transversal para estimar la prevalencia de asma ocupacional en la población laboral expuesta del municipio Guantánamo durante el período de julio del 2000 a febrero del 2001. El universo de estudio estuvo integrado por 838 trabajadores pertenecientes a la industria sideromecánica y a la construcción; se trabajó con una muestra de 189 trabajadores, quienes fueron seleccionados a través de un muestreo estratificado polietápico. Se encontró que el mayor numero de trabajadores corresponden al sexo masculino, siendo la exposición ocupacional a polvos, y el tiempo de exposición elementos significativos de predicción de asma. Los síntomas y signos encontrados en la serie de estudio, en orden de frecuencia corresponden a la hiperreactividad bronquial con 58.3 (por ciento), tos 25.0 (por ciento), disnea 14.2 (por ciento) y sibilancia 2.3 (por ciento), asociándose la disfunción ventilatoria obstructiva reversible en 52.3 (por ciento) y los resultados positivos del prick- test en 66.7 (por ciento) de los individuos estudiados. Esta última constituye un elemento significativo de predicción de asma. Se detectan, según criterios de Brook, 17 trabajadores con diagnóstico de asma ocupacional, lo que representa una tasa general de 2 (por ciento)(AU)