Deletion Mutants of the Attenuated Recombinant ASF Virus, BA71ΔCD2, Show Decreased Vaccine Efficacy.

African swine fever (ASF) has become the major threat to the global swine industry. Lack of available commercial vaccines complicates the implementation of global control strategies. So far, only live attenuated ASF viruses (ASFV) have demonstrated solid protection efficacy at the experimental level. The implementation of molecular techniques has allowed the generation of a collection of deletion mutants lacking ASFV-specific virulence factors, some of them with promising potential as vaccine candidates against the pandemic genotype II ASFV strain currently circulating in Africa, Europe, Asia and Oceania. Despite promising results, there is room for improvement, mainly from the biosafety point of view. Aiming to improve the safety of BA71∆CD2, a cross-protective recombinant live attenuated virus (LAV) lacking the ASFV CD2v gene (encoding ß-glucuronidase as a reporter gene) available in our laboratory, three new recombinants were generated using BA71∆CD2 as a template: the single mutant BA71∆CD2f, this time containing the fluorescent mCherry reporter gene instead of CD2v, and two double recombinants lacking CD2v and either the lectin gene (EP153R) or the uridine kinase (UK) gene (DP96R). Comparative in vivo experiments using BA71∆CD2f, BA71∆CD2DP96R and BA71∆CD2EP153R recombinant viruses as immunogens, demonstrated that deletion of either DP96R or EP153R from BA71∆CD2f decreases vaccine efficacy and does not improve safety. Our results additionally confirm ASFV challenge as the only available method today to evaluate the protective efficacy of any experimental vaccine. We believe that understanding the fine equilibrium between attenuation and inducing protection in vivo deserves further study and might contribute to more rational vaccine designs in the future.

M448R and MGF505-7R: Two African Swine Fever Virus Antigens Commonly Recognized by ASFV-Specific T-Cells and with Protective Potential.

African swine fever (ASF) is today's number one threat for the global swine industry. Neither commercial vaccine nor treatment is available against ASF and, thus far, only live attenuated viruses (LAV) have provided robust protection against lethal ASF virus (ASFV) challenge infections. Identification of ASFV proteins inducing protective immune responses is one of the major challenges to develop safer and efficient subunit vaccines. Immunopeptidomic studies recently performed in our laboratory allowed identifying ASFV antigens recognized by ASFV-specific CD8+ T-cells. Here, we used data from the SLAI-peptide repertoire presented by a single set of ASFV-infected porcine alveolar macrophages to generate a complex DNA vaccine composed by 15 plasmids encoding the individual peptide-bearing ORFs. DNA vaccine priming improved the protection afforded by a suboptimal dose of the BA71ΔCD2 LAV given as booster vaccination, against Georgia2007/1 lethal challenge. Interestingly, M448R was the only protein promiscuously recognized by the induced ASFV-specific T-cells. Furthermore, priming pigs with DNA plasmids encoding M488R and MGF505-7R, a CD8+ T-cell antigen previously described, confirmed these two proteins as T-cell antigens with protective potential. These studies might be useful to pave the road for designing safe and more efficient vaccine formulations in the future.

Identification of Promiscuous African Swine Fever Virus T-Cell Determinants Using a Multiple Technical Approach.

The development of subunit vaccines against African swine fever (ASF) is mainly hindered by the lack of knowledge regarding the specific ASF virus (ASFV) antigens involved in protection. As a good example, the identity of ASFV-specific CD8+ T-cell determinants remains largely unknown, despite their protective role being established a long time ago. Aiming to identify them, we implemented the IFNγ ELISpot as readout assay, using as effector cells peripheral blood mononuclear cells (PBMCs) from pigs surviving experimental challenge with Georgia2007/1. As stimuli for the ELISpot, ASFV-specific peptides or full-length proteins identified by three complementary strategies were used. In silico prediction of specific CD8+ T-cell epitopes allowed identifying a 19-mer peptide from MGF100-1L, as frequently recognized by surviving pigs. Complementarily, the repertoire of SLA I-bound peptides identified in ASFV-infected porcine alveolar macrophages (PAMs), allowed the characterization of five additional SLA I-restricted ASFV-specific epitopes. Finally, in vitro stimulation studies using fibroblasts transfected with plasmids encoding full-length ASFV proteins, led to the identification of MGF505-7R, A238L and MGF100-1L as promiscuously recognized antigens. Interestingly, each one of these proteins contain individual peptides recognized by surviving pigs. Identification of the same ASFV determinants by means of such different approaches reinforce the results presented here.

Live Attenuated African Swine Fever Viruses as Ideal Tools to Dissect the Mechanisms Involved in Cross-Protection.

African swine fever (ASF) has become the major threat for the global swine industry. Furthermore, the epidemiological situation of African swine fever virus (ASFV) in some endemic regions of Sub-Saharan Africa is worse than ever, with multiple virus strains and genotypes currently circulating in a given area. Despite the recent advances on ASF vaccine development, there are no commercial vaccines yet, and most of the promising vaccine prototypes available today have been specifically designed to fight the genotype II strains currently circulating in Europe, Asia, and Oceania. Previous results from our laboratory have demonstrated the ability of BA71∆CD2, a recombinant LAV lacking CD2v, to confer protection against homologous (BA71) and heterologous genotype I (E75) and genotype II (Georgia2007/01) ASFV strains, both belonging to same clade (clade C). Here, we extend these results using BA71∆CD2 as a tool trying to understand ASFV cross-protection, using phylogenetically distant ASFV strains. We first observed that five out of six (83.3%) of the pigs immunized once with 106 PFU of BA71∆CD2 survived the tick-bite challenge using Ornithodoros sp. soft ticks naturally infected with RSA/11/2017 strain (genotype XIX, clade D). Second, only two out of six (33.3%) survived the challenge with Ken06.Bus (genotype IX, clade A), which is phylogenetically more distant to BA71∆CD2 than the RSA/11/2017 strain. On the other hand, homologous prime-boosting with BA71∆CD2 only improved the survival rate to 50% after Ken06.Bus challenge, all suffering mild ASF-compatible clinical signs, while 100% of the pigs immunized with BA71∆CD2 and boosted with the parental BA71 virulent strain survived the lethal challenge with Ken06.Bus, without almost no clinical signs of the disease. Our results confirm that cross-protection is a multifactorial phenomenon that not only depends on sequence similarity. We believe that understanding this complex phenomenon will be useful for designing future vaccines for ASF-endemic areas.

Working Conditions, Workplace Violence, and Psychological Distress in Andean Miners: A Cross-sectional Study Across Three Countries.

BACKGROUND: Psychosocial working conditions are well-known determinants of poor mental health. However, studies in mining populations where employment and working conditions are frequently precarious have, to our knowledge, only focused on occupational accidents and diseases. OBJECTIVES: The aim of this study was to assess psychosocial working conditions and psychological distress in Andean underground miners. METHODS: The study population consisted of 153 Bolivian miners working in a silver mining cooperative, 137 Chilean informal gold miners, and 200 formal Peruvian silver miners employed in a remote setting. High work demands, minimal work control, minimal social support at work, and workplace exposure to violence and bullying were assessed using the Spanish short form of the European Working Condition Survey. A general health questionnaire score >4 was used as cutoff for psychological distress. Associations between psychosocial work environment and psychological distress were tested using logistic regression models controlling for potential confounding and effect modification by country. FINDINGS: Prevalence of psychological distress was 82% in the Bolivian cooperative miners, 29% in the Peruvian formal miners, and 22% in the Chilean informal miners (pχ(2) < 0.001). 55% of the miners had suffered violence during the 12-months before the survey. Workplace demands were high (median 12.5 on a scale from 7-14), as was social support (median 5.5 on a scale from 3-6). After adjustment for country and other relevant exposure variables and considering interactions between country and job strain, miners in active (odds ratio [OR], 6.8; 95% confidence interval [CI] 2.1-22.7) and high strain jobs (OR, 7.2; 95% CI, 1.7-29.9) were at increased odds of distress compared with those in low strain jobs. Violence at work also contributed to increased odds of distress (OR, 1.86; 95% CI, 1.1-3.1). CONCLUSIONS: Psychological distress is associated with the psychosocial work environment in Andean underground miners. Interventions in mining populations should take the psychosocial work environment into account.

Utilidad del Índice de Pronóstico Inflamatorio y Nutricional (PINI) en el diagnóstico de niños con desnutrición grave / Usefulness of Inflammatory Prognostic Index and Nutrition (PINI) in the diagnosis of children with severe malnutrition

Objetivos: evaluar el Índice de Pronóstico Inflamatorio Nutricional (PINI) mediante la determinación de Albúmina, Prealbúmina, Alfa-1-Glicoproteína y Proteína C Reactiva como pronóstico de riesgo durante la rehabilitación nutricional de niños desnutridos graves. Métodos: se estudiaron 36 niños(as) desnutridos infectados hospitalizados en el CRIN y 30 niños(as) controles entre 6 meses y 5 años con valoración antropométrica y nutricional. Se determinaron parámetros hematológicos y proteínas nutricionales e inflamatorias por turbi-dimetría. Los resultados se expresaron en términos de media ± desviación estándar con análisis de varianza en una sola dirección (ANO-VA) y la t de Student. Con 95% de confiabilidad estadística (p < 0,05) con el paquete estadístico STATA v. 10. Resultados: la antropometría mostró diferencia significativa en todos los parámetros comparados con el grupo control y umbral de referencia del National Center for Health Statistics. Parámetros hematológicos por debajo del límite inferior de la normalidad. Sesenta y nueve por ciento con anemia, 19 niños presentaron déficit de transferrina, 25% leucocitosis, 8% eosinofilia, 14% neutrofilia y 14% parasitosis. Las concentraciones de albúmina y prealbúmina significativamente disminuidas, la Alfa-1-Glicoproteína Ácida y Proteína C Reactiva, significativamente superiores a los niños control (p<0,001). Veinticinco por ciento presentaron un PINI de riesgo débil, 42% riesgo moderado, 22% riesgo alto y 11% riesgo vital correlacionado con patologías asociadas. Los niños sanos, un PINI menor a 1. Conclusiones: el PINI como complemento de la clínica ayuda a valorar la gravedad inicial de la infección, a preverla y a seguir su evolución en la desnutrición infantil. Es un micrométodo simple, rápido que puede ser repetido regularmente.

Objectives: To evaluate the prognostic inflammatory and nutritional index (PINI) by determining Albumin, Prealbumin, Alpha-1-glyco-protein and C-reactive protein as a predictor of risk for nutritional rehabilitation of severely malnourished children. Methods: We studied 36 children (as) in hospitalized malnourished CRIN infected and 30 children (as) controls between 6 months and 5 years with anthropo-metric and nutritional assessment. Hematological parameters were determined, and nutritional and inflammatory proteins by turbidi-metry. The results were expressed as mean ± standard deviation with ANOVA one-way and Student's t test. 95% statistical confidence (P <0.05) with the statistical package STATA. 10. Results: Anthropometry showed a significant difference in all parameters compared with the control group and reference threshold of the National Center for Health Statistics. Hematologic parameters below the lower limit of normality. 69% with anemia, 19 were deficient transferrin, leukocytosis 25%, 8% eosinophilia, 14% neutrophils and 14% parasitized. The albumin and prealbumin concentrations significantly decreased, the Alpha-1-acid glycoprotein and C-reactive protein significantly hig-her than control children (p<0.001). 25% showed a weak risk PINI, 42% moderate risk, high risk 22% and 11% correlated with life-threa-tening comorbidities. Healthy children, a PINI less than 1. Conclusions: The PINI as a complement to the clinic helps to assess the initial severity of the infection, to foresee and monitor trends in child malnutrition. A micromethod is simple, fast as it can be repeated regularly.

Siringoma eruptivo familiar / Familial syringoma

Los siringomas eruptivos son dermatosis papulares relativamente raras que afectan la superficie anterior del cuerpo. Surgen comúnmente en la infancia, con un curso benigno, eventualmente regresionan y presentan pobre respuesta a los tratamientos. La forma de siringomas eruptivos familiares es una manifestación aún más rara. Se acoseja considerarlos entre los diagnósticos diferenciales cuando se observen lesiones papulares en niños, a fin de que se les identifique y los tratamientos sean los mas apropiados. Presentamos tres casos adicionales de siringoma eruptivo.

Hamartoma angiomatoso ecrino. Reporte de un caso y revisión de la literatura / Eccrine angiomatous hamartoma. Report of a case and literature review

El Hamartoma angiomatoso ecrino es una rara lesión cutánea benigna caracteirzada histológicmaente por la proliferación de glándulas ecrinas asociadas cercanamente a un estroma vascular. Típicamente es descrita como una tumoración de aspecto angiomatoso de tamaño creciente en una extremidad, dolorosa, hiperhidrótica e hipertricótica, que aparece al nacimiento o durante la infancia. Presentamos un caso en una niña de 5 años de edad que reunió todas estas características que nos ayudaron a diferenciarlo de otras anormalidades vasculares y que fue tratada quirúrgicamente con éxito.

Tiña negra palmar. Reporte de dos casos / Tinea nigra. Report of two cases

La tiña negra es una dermatomicosis superficial causada por la Phaeoannellomyces (Exophiala) werneckii. Reportamos dos casos de afectación palmar en pacientes procedentes de la selva alta del Perú. El examen directo y el cultivo confirmaron el diagnóstico. El tratamiento con imidazólicos tópicos fue suficiente y exitoso.