Theranostic applications of phage display to control leishmaniasis: selection of biomarkers for serodiagnostics, vaccination, and immunotherapy

AbstractPhage display is a high-throughput subtractive proteomic technology used for the generation and screening of large peptide and antibody libraries. It is based on the selection of phage-fused surface-exposed peptides that recognize specific ligands and demonstrate desired functionality for diagnostic and therapeutic purposes. Phage display has provided unmatched tools for controlling viral, bacterial, fungal, and parasitic infections, and allowed identification of new therapeutic targets to treat cancer, metabolic diseases, and other chronic conditions. This review presents recent advancements in serodiagnostics and prevention of leishmaniasis -an important tropical parasitic disease- achieved using phage display for the identification of novel antigens with improved sensitivity and specificity. Our focus is on theranostics of visceral leishmaniasis with the aim to develop biomarker candidates exhibiting both diagnostic and therapeutic potential to fight this important, yet neglected, tropical disease.

Impact of carbon/nitrogen feeding strategy on polyhydroxyalkanoates production using mixed microbial cultures

Polyhydroxyalkanoates (PHA) production using mixed microbial cultures (MMC) requires a multi-stage process involving the microbial selection of PHA-storing microorganisms, typically operated in sequencing batch reactors (SBR), and an accumulation reactor. Since low-cost renewable feedstocks used as process feedstock are often nitrogen-deficient, nutrient supply in the selection stage is required to allow for microbial growth. In this context, the possibility to uncouple nitrogen supply from carbon feeding within the SBR cycle has been investigated in this study. Moreover, three different COD:N ratios (100:3.79, 100:3.03 and 100:2.43) were tested in three different runs which also allowed the study of COD:N ratio on the SBR performance. For each run, a synthetic mixture of acetic and propionic acids at an overall organic load rate of 8.5 gCOD L-1 d-1 was used as carbon feedstock, whereas ammonium sulfate was the nitrogen source in a lab-scale sequence batch reactor (SBR) with 1 L of working volume. Besides, a sludge retention time (SRT) of 1 d was used as well as a 6 h cycle length. The uncoupled feeding strategy significantly enhanced the selective pressure towards PHA-storing microorganisms, resulting in a two-fold increase in the PHA production (up to about 1.3 gCOD L-1). A high storage response was observed for the two runs with the COD:N ratios (gCOD:gN) of 100:3.79 and 100:3.03, whereas the lowest investigated nitrogen load resulted in very poor performance in terms of polymer production. In fact, strong nitrogen limitation caused fungi to grow and a very poor storage ability by microorganisms that thrived in those conditions. The COD:N ratio also affected the polymer composition, indeed the produced poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) showed a variable HV content (1-20 %, w/w) among the three runs, lessening as the COD:N increased. This clearly suggests the possibility to use the COD:N ratio as a tool for tuning polymer properties regardless the composition of the feedstock.

Acute disseminated encephalomyelitis following inactivated influenza vaccination in the Brazilian Amazon: a case report

AbstractHere, we describe a case of acute disseminated encephalomyelitis (ADEM) that occurred during a plausible risk interval following inactivated influenza vaccination in a previously healthy 27-year-old man from Manaus, Brazil. He was treated with intravenous methylprednisolone and immunoglobulin. One-month follow-up revealed resolution of the brain lesions, but not of the spinal cord lesions. No recurrence or progression of the main neurological symptoms was observed. After two years of monitoring, the patient continues to experience weak lower limbs and urinary retention. Thus, we recommend that ADEM should be considered in a patient presenting with neurological symptoms after influenza vaccination.

Risk-benefit assessment of Bacillus Calmette-Guérin vaccination, anti-phenolic glycolipid I serology, and Mitsuda test response: 10-year follow-up of household contacts of leprosy patients

Abstract: INTRODUCTION: Despite multidrug therapy, leprosy remains a public health issue. The intradermal Bacillus Calmette-Guérin (BCG) vaccine, Mitsuda test (lepromin skin test), and anti-phenolic glycolipid I (PGL-I) serology are widely used in leprosy studies and have shown great epidemiological value. METHODS: This longitudinal study evaluated the relative risks and benefits of these three tools by comparing results observed in household contacts (HHCs) of leprosy patients who developed leprosy with those of HHCs who did not in a population of 2,992 individuals monitored during a 10-year period. RESULTS : Seventy-five (2.5%) new leprosy cases were diagnosed, including 28 (0.9%) co-prevalent cases. Therefore, for the risk-benefit assessment, 47 (1.6%) HHCs were considered as truly diagnosed during follow-up. The comparison between healthy and affected contacts demonstrated that not only did BCG vaccination increase protection, but boosters also increased to 95% relative risk (RR) reduction when results for having two or more scars were compared with having no scars [RR, 0.0459; 95% confidence interval (CI), 0.006-0.338]. Similarly, Mitsuda reactions >7mm in induration presented 7-fold greater protection against disease development compared to reactions of 0-3mm (RR, 0.1446; 95% CI, 0.0566-0.3696). In contrast, anti-PGL-I ELISA seropositivity indicated a 5-fold RR increase for disease outcome (RR, 5.688; 95% CI, 3.2412-9.9824). The combined effect of no BCG scars, Mitsuda reaction of <7mm, and seropositivity to anti-PGL-I increased the risk for leprosy onset 8-fold (RR, 8.109; 95% CI, 5.1167-12.8511). CONCLUSIONS: The adoption of these combined assays may impose measures for leprosy control strategies.

Epidemiology of infectious meningitis in the State of Amazonas, Brazil

INTRODUCTION: In the State of Amazonas, particularly in the capital Manaus, meningitis has affected populations of different cultures and social strata over the years. Bacterial meningitis is caused by several different species and represents a major issue of public health importance. The present study reports the meningitis case numbers with different etiologies in Amazonas from January 1976 to December 2012. METHODS: Since the 1970s, the (currently named) Tropical Medicine Foundation of Doutor Heitor Vieira Dourado [Fundação de Medicina Tropical Doutor Heitor Vieira Dourado (FMT-HVD)] has remained a reference center in Amazonas for the treatment of meningitis through the diagnosis and notification of cases and the confirmation of such cases using specific laboratory tests. RESULTS: The foundation has achieved coverage of over 90% of the state medical records for many years. Between 1990 and 2012, meningitis cases caused by Haemophilus influenzae decreased with the introduction of the H. influenzae vaccine. Meningococcal disease previously had a higher frequency of serogroup B disease, but starting in 2008, the detection of serogroup C increased gradually and has outpaced the detection of serogroup B. Recently, surveillance has improved the etiological definition of viral meningitis at FMT-HVD, with enteroviruses, Epstein-Barr virus (EBV) and varicella zoster virus (VZV) prevailing in this group of pathogens. With the advent of acquired immunodeficiency syndrome (AIDS), cryptococcal meningitis has become an important disease in Amazonas. Additionally, infectious meningitis is an important burden in the State of Amazonas. CONCLUSIONS: Changes in the epidemiological profile for the different etiology-defined cases are the result of continuous epidemiological surveillance and laboratory capacity improvements and control measures, such as Haemophilus influenzae vaccination.

Prevalence and genotype distribution of human papillomavirus: implications for cancer screening and vaccination in Henan province, China

Abstract: INTRODUCTION: To provide information for cervical cancer screening and vaccination in Henan province, China, the distribution of human papillomavirus (HPV) was analyzed. METHODS: The HPV genotypes were detected using gene array and flow-through hybridization. RESULTS: Overall, 38.1% (1,536/4,033) of the women were human papillomavirus deoxyribonucleic acid (HPV DNA) positive. The prevalence of high-risk HPV types was 32.4%. HPV 16 was the most prevalent genotype (8.9%), followed by HPV 52 (5.8%) and HPV 58 (4.4%). CONCLUSIONS: The data support close surveillance of women for cervical cancer screening, and HPV prophylactic vaccines including HPV16, HPV 52, and HPV 58 might offer greater protection in this area.

Polyhydroxyalkanoates production by photosynthetic mixed cultures

Polyhydroxyalkanoates (PHAs) are natural biologically synthesized polymers that have been the subject of much interest in the last decades due to their biodegradability. Thus far, its microbial production is associated with high operational costs, which increases PHA prices and limits its marketability. To address this situation, this thesis’ work proposes the utilization of photosynthetic mixed cultures (PMC) as a new PHA production system that may lead to a reduction in operational costs. In fact, the operational strategies developed in this work led to the selection of PHA accumulating PMCs that, unlike the traditional mixed microbial cultures, do not require aeration, thus permitting savings in this significant operational cost. In particular, the first PHA accumulating PMC tested in this work was selected under non-aerated illuminated conditions in a feast and famine regime, being obtained a consortium of bacteria and algae, where photosynthetic bacteria accumulated PHA during the feast phase and consumed it for growth during the famine phase, using the oxygen produced by algae. In this symbiotic system, a maximum PHA content of 20% cell dry weight (cdw) was reached, proving for the first time, the capacity of a PMC to accumulate PHA. During adaptation to dark/light alternating conditions, the culture decreased its algae content but maintained its viability, achieving a PHA content of 30% cdw. Also, the PMC was found to be able to utilize different volatile fatty acids for PHA production, accumulating up to 20% cdw of a PHA co-polymer composed of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (HV) monomers. Finally, a new selective approach for the enrichment of PMCs in PHA accumulating bacteria was tested. Instead of imposing a feast and famine regime, a permanent feast regime was used, thus selecting a PMC that was capable of simultaneously growing and accumulating PHA, being attained a maximum PHA content of 60% cdw, the highest value reported for a PMC thus far. The results presented in this thesis prospect the utilization of cheap, VFA-rich fermented wastes as substrates for PHA production, which combined with this new photosynthetic technology opens up the possibility for direct sunlight illumination, leading to a more cost-effective and environmentally sustainable PHA production process.

Effects of residues on the degradation of PHA produced from mixed microbial cultures and processed in extrusion

Thermal degradation upon melting is one of the major drawbacks reported for polyhydroxyalkanoates (PHA). However, the role of residues originating from the fermentation and the extraction steps on the thermal stability of this class of biopolymers still needs to be clarified. In the particular case of PHA produced from mixed microbial cultures (MMC), this topic is even less documented in the literature. Here, two polyhydroxy(butyrate-co-valerate) (PHBV) produced from MMC enriched in PHA accumulating organisms and fed with cheese whey were studied. A micro extrusion line is used to produce filaments and assess the processability and the degradation of processed PHBV. The prototype micro extrusion line allows for studying grams of materials. The two PHBV contain 18 mol% HV. PHBV was recovered with 11 wt% residues, and further submitted to a purification procedure resulting in a second biopolyester containing less than 2 wt% impurities. The thermorheological characterization of the two PHBV is first presented, together with their semicrystalline properties. Then the processing windows of the two biopolyesters are presented. Finally, the properties of extruded filaments are reported and the thermomechanical degradation of PHBV is extensively studied. The structure was assessed by wide angle X-ray diffraction, mechanical and rheological properties are reported, thermal properties are studied with differential scanning calorimetry and thermogravimetric analysis, whereas Fourier Transform Infrared spectroscopy was used to assess the impact of the extrusion on PHBV chemical structure. All results obtained with the two PHBV are compared to assess the effects of residues on both PHBV processability and degradation.

Local seismic cultures: the use of timber frame structures in the south of Portugal

Given the fact that using timber frame structures has proven to improve the seismic behavior of vernacular architecture, as has been reported in past earthquakes in many countries, its preservation as a traditional earthquake resistant practice is important. This paper firstly intends to evaluate whether the use of timber frames as a traditional seismic resistant technique for vernacular architecture in the South of Portugal, traditionally a seismic region, is still active. Secondly, the city of Vila Real de Santo António was selected as a case study because it also followed a Pombaline development contemporary to the reconstruction of Lisbon. The plan included the provision of timber frame partition walls for some of the buildings and, thus, an overview of the type of constructions originally conceived is provided. Finally, the alterations done in the original constructions and the current state of the city center are described and the effect of these changes on the seismic vulnerability of the city is discussed.

Effect of fetal bovine serum on Candida glabrata cultures

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica)

Production of secondary metabolites in cell suspension cultures of Ocimum sanctum L.

Dissertação de mestrado em Plant Molecular Biology, Biotechnology and Bioentrepreneurship

Effects of fermentation residues on the melt processability and thermomechanical degradation of PHBV produced from cheese whey using mixed microbial cultures

"Available online 28 March 2016"

Proyecto Fortalecimiento de los Gobiernos Locales y de las ONG´s para el desarrollo Medio y Socioambiental en la Región de Punilla en la Provincia de Córdoba

El presente proyecto plantea la profundización de una línea de investigación ya existente en el ICDA que cuenta con apoyo del VRMU de la UCC y aval de la SECyT, sobre la identificación de problemas y necesidades de los gobiernos locales y las ONG´s de la Región de Punilla de la Provincia de Córdoba en torno a la problemática ambiental y la promoción de prácticas participativas para su abordaje y resolución. Este proyecto tiene como objetivo inmediato promover el fortalecimiento de los Gobiernos locales y de las ONGs, siguiendo las líneas de trabajo del ICDA a través de sus centros especializados CIPEAP–Profim y CIPAC–Compás respectivamente, en lo referido a gestión sustentable del medio ambiente y la utilización y/o conservación de los recursos naturales, principalemente el agua. La investigación procura identificar las áreas problema en el marco de la gestión socio-ambiental focalizada en la gestión estratégica del recurso hídrico con el fin de sistematizar los aspectos que facilitan u obstaculizan el acuerdo, la gestión asociada y la administración eficiente de los recursos naturales en la región. Entre las necesidades prioritarias vinculadas a la gestión socio – ambiental se delimitaron los núcleos del problema de la gestión estratégica del recurso hídrico, como principal eje de desarrollo de la región. En este sentido Punilla enfrenta diferentes niveles de problemáticas; en primer lugar, se encuentra la contaminación de la principal cuenca hídrica que provee agua potable para consumo humano a los mayores centros urbanos con consecuencias directas sobre la salud ambiental; de esta manera el saneamiento y las metodologías a aplicarse, representan un espacio de confrontación entre los actores tanto gubernamentales como no gubernamentales por implementar prácticas de saneamiento sin una visión de desarrollo integral. En segundo lugar la región enfrenta la amenaza de la escasez del recurso, como consecuencia de la creciente urbanización y de las barreras geográficas que en muchos casos impiden obras de infraestructura que garanticen el acceso igualitario al mismo. Finalmente las consecuencias y efectos desde el punto de vista de la salud de las personas junto a la disminución de oportunidades y los problemas del desarrollo socio-económico de la región, representan un tercer eje de investigación en consonancia con las Políticas y Estrategias de Investigación de la UCC (RR 492/07). Dada la complejidad del proyecto, resulta fundamental el apoyo que brinda la Escuela Andaluza de Salud Pública (EASP), como socia en la presentación a la convocatoria realizada para obtener el financiamento por parte de la Agencia Española de Cooperación Internacional para el Desarrollo (AECID).

Factores críticos de desarrollo de comunidades locales y regionales de la zona pampeana de la Provincia de Córdoba

El proyecto es la primera de tres etapas previstas para determinar factores críticos que inciden en el desarrollo local y regional integrado en la Provincia de Córdoba. Las poblaciones del interior de la Provincia de Córdoba, con exclusión de la Capital y de las principales ciudades del interior, han padecido los procesos de concentración geográfica del ingreso, la riqueza, los presupuestos oficiales y las migraciones. Muchas veces librada a su suerte, la sociedad civil de estas comunidades desarrolló y capitalizó una red de instituciones que prestan los servicios que el Estado no cubrió. Los gobiernos locales, a la vez, intentan, con éxitos limitados, detener el procesos de descapitalización social -financiera y humana de sus comunidades. Las empresas, con ciclos recurrentes de crecimiento y decrecimiento vinculados a los mercados, se debaten en la búsqueda de los escasos recursos, financieros y humanos, que les permitan consolidar un desarrollo a mediano y largo plazo. Juntos tienen, especialmente en la región pampeana de la Provincia, una nueva oportunidad de hacerlo. Según nuestra hipótesis, incidirían en estos procesos como factores críticos: 1. Aspectos exógenos que descapitalizan las comunidades como los tributarios (impuestos y gasto geográfico), financieros (colocaciones y préstamos), educativos (migración de los miembros más capacitados), sanitarios (por la concentración de las prestaciones de mayor complejidad), y de prestación de servicios públicos (por las mayores tarifas y menores prestaciones) de las empresas de capital concentrado. 2. Aspectos endógenos que producen des-economías, ineficiencias y desaprovechamiento de los recursos (humanos, productivos, tecnológicos, organizativos y financieros) locales y regionales, como: la falta de un proyecto local y/o regional común; dinámicas no participativas; insuficiencia, dispersión y desconocimiento de información clave para la planeación; diferencias personales y conflictos de roles; carencia de cadenas productivas integradas verticalmente El proyecto propone como objetivos: 1. Identificar y, en la medida de lo posible, cuantificar los factores críticos exógenos y endógenos para el desarrollo local y regional integrado; 2. elaborar sobre esta base modelos de desarrollo, proyectos interinstitucionales y propuestas de cadenas productivas que disminuyan los problemas y potencien las oportunidades de las comunidades regionales estudiadas