Building Programs to Eradicate Toxoplasmosis Part I: Introduction and Overview.

Purpose of Review: Review building of programs to eliminate Toxoplasma infections. Recent Findings: Morbidity and mortality from toxoplasmosis led to programs in USA, Panama, and Colombia to facilitate understanding, treatment, prevention, and regional resources, incorporating student work. Summary: Studies foundational for building recent, regional approaches/programs are reviewed. Introduction provides an overview/review of programs in Panamá, the United States, and other countries. High prevalence/risk of exposure led to laws mandating testing in gestation, reporting, and development of broad-based teaching materials about Toxoplasma. These were tested for efficacy as learning tools for high-school students, pregnant women, medical students, physicians, scientists, public health officials and general public. Digitized, free, smart phone application effectively taught pregnant women about toxoplasmosis prevention. Perinatal infection care programs, identifying true regional risk factors, and point-of-care gestational screening facilitate prevention and care. When implemented fully across all demographics, such programs present opportunities to save lives, sight, and cognition with considerable spillover benefits for individuals and societies. Supplementary Information: The online version contains supplementary material available at 10.1007/s40124-022-00269-w.

Building Programs to Eradicate Toxoplasmosis Part IV: Understanding and Development of Public Health Strategies and Advances "Take a Village".

Purpose of Review: Review international efforts to build a global public health initiative focused on toxoplasmosis with spillover benefits to save lives, sight, cognition and motor function benefiting maternal and child health. Recent Findings: Multiple countries' efforts to eliminate toxoplasmosis demonstrate progress and context for this review and new work. Summary: Problems with potential solutions proposed include accessibility of accurate, inexpensive diagnostic testing, pre-natal screening and facilitating tools, missed and delayed neonatal diagnosis, restricted access, high costs, delays in obtaining medicines emergently, delayed insurance pre-approvals and high medicare copays taking considerable physician time and effort, harmful shortcuts being taken in methods to prepare medicines in settings where access is restricted, reluctance to perform ventriculoperitoneal shunts promptly when needed without recognition of potential benefit, access to resources for care, especially for marginalized populations, and limited use of recent advances in management of neurologic and retinal disease which can lead to good outcomes. Supplementary Information: The online version contains supplementary material available at 10.1007/s40124-022-00268-x.

Comparison of PvLAP5 and Pvs25 qRT-PCR assays for the detection of Plasmodium vivax gametocytes in field samples preserved at ambient temperature from remote malaria endemic regions of Panama.

BACKGROUND: As the elimination of malaria in Mesoamerica progresses, detection of Plasmodium vivax using light microscopy (LM) becomes more difficult. Highly sensitive molecular tools have been developed to help determine the hidden reservoir of malaria transmission in low transmission settings. In this study we compare the performance of PvLAP5 and Pvs25 qRT-PCR assays to LM for the detection of Plasmodium vivax gametocytes in field samples preserved at ambient temperature from malaria endemic regions of Panama. METHODS: For this purpose, we collected a total of 83 malaria field samples during 2017-2020 preserved in RNAprotect (RNAp) of which 63 (76%) were confirmed P. vivax by LM and selected for further analysis. Additionally, 16 blood samples from local healthy malaria smear negative volunteers, as well as, from 15 malaria naïve lab-bred Aotus monkeys were used as controls. To optimize the assays, we first determined the minimum blood volume sufficient for detection of PvLAP5 and Pv18SrRNA using P. vivax infected Aotus blood that was preserved in RNAp and kept either at ambient temperature for up to 8 days before freezing or was snap-frozen at -80° Celsius at the time of bleeding. We then compared the mean differences in gametocyte detection rates of both qRT-PCR assays to LM and performed a multivariate correlation analysis of study variables. Finally, we determined the sensitivity (Se) and specificity (Sp) of the assays at detecting gametocytes compared to LM. RESULTS: Blood volume optimization indicated that a blood volume of at least 60 µL was sufficient for detection of PvLAP5 and Pv18SrRNA and no significant differences were found between RNA storage conditions. Both PvLAP5 and Pvs25 qRT-PCR assays showed a 37-39% increase in gametocyte detection rate compared to LM respectively. Strong positive correlations were found between gametocytemia and parasitemia and both PvLAP5 and Pvs25 gametocyte markers. However, no significant differences were detected in the Se and Sp of the Pvs25 and PvLAP5 qRT-PCR assays, even though data from control samples suggested Pvs25 to be more abundant than PvLAP5. CONCLUSIONS: This study shows that the PvLAP5 qRT-PCR assay is as Se and Sp as the gold standard Pvs25 assay and is at least 37% more sensitive than LM at detecting P. vivax gametocytes in field samples preserved in RNAp at ambient temperature from malaria endemic regions of Panama. AUTHOR SUMMARY: Plasmodium vivax is one of the five species of malaria (P. falciparum, P. malariae, P. ovale and P. knowlesi) that are transmitted to man by the bite of female anopheles mosquitoes. It causes ~14.3 million cases mainly in Southeast Asia, India, the Western Pacific and the Americas annually. In the Americas, malaria remains a major problem in underdeveloped areas and indigenous communities in the Amazon region and eastern Panama, where it is endemic and difficult to eliminate. As malaria elimination progresses, detection of P. vivax by light microscopy (LM) becomes more difficult. Therefore, highly sensitive molecular tools have been developed that use genetic markers for the parasite to help determine the hidden reservoir of malaria transmission. This study compares the performance of two molecular assays based on the genetic markers of mature gametocytes PvLAP5 and Pvs25 with LM. The study shows that the PvLAP5 qRT-PCR assay is as sensitive and specific as the gold standard Pvs25 assay and is at least 37% more sensitive than LM at detecting P. vivax gametocytes. These data suggest that the PvLAP5 qRT-PCR assay can be a useful tool to help determine the hidden reservoir of transmission in endemic foci approaching elimination.

Building Programs to Eradicate Toxoplasmosis Part II: Education.

Purpose of Review: Review work to create and evaluate educational materials that could serve as a primary prevention strategy to help both providers and patients in Panama, Colombia, and the USA reduce disease burden of Toxoplasma infections. Recent Findings: Educational programs had not been evaluated for efficacy in Panama, USA, or Colombia. Summary: Educational programs for high school students, pregnant women, medical students and professionals, scientists, and lay personnel were created. In most settings, short-term effects were evaluated. In Panama, Colombia, and USA, all materials showed short-term utility in transmitting information to learners. These educational materials can serve as a component of larger public health programs to lower disease burden from congenital toxoplasmosis. Future priorities include conducting robust longitudinal studies of whether education correlates with reduced adverse disease outcomes, modifying educational materials as new information regarding region-specific risk factors is discovered, and ensuring materials are widely accessible.

Building Programs to Eradicate Toxoplasmosis Part III: Epidemiology and Risk Factors.

Purpose of Review: Review comprehensive data on rates of toxoplasmosis in Panama and Colombia. Recent Findings: Samples and data sets from Panama and Colombia, that facilitated estimates regarding seroprevalence of antibodies to Toxoplasma and risk factors, were reviewed. Summary: Screening maps, seroprevalence maps, and risk factor mathematical models were devised based on these data. Studies in Ciudad de Panamá estimated seroprevalence at between 22 and 44%. Consistent relationships were found between higher prevalence rates and factors such as poverty and proximity to water sources. Prenatal screening rates for anti-Toxoplasma antibodies were variable, despite existence of a screening law. Heat maps showed a correlation between proximity to bodies of water and overall Toxoplasma seroprevalence. Spatial epidemiological maps and mathematical models identify specific regions that could most benefit from comprehensive, preventive healthcare campaigns related to congenital toxoplasmosis and Toxoplasma infection.

Population genomics of Plasmodium vivax in Panama to assess the risk of case importation on malaria elimination.

Malaria incidence in Panama has plateaued in recent years in spite of elimination efforts, with almost all cases caused by Plasmodium vivax. Notwithstanding, overall malaria prevalence remains low (fewer than 1 case per 1000 persons). We used selective whole genome amplification to sequence 59 P. vivax samples from Panama. The P. vivax samples were collected from two periods (2007-2009 and 2017-2019) to study the population structure and transmission dynamics of the parasite. Imported cases resulting from increased levels of human migration could threaten malaria elimination prospects, and four of the samples evaluated came from individuals with travel history. We explored patterns of recent common ancestry among the samples and observed that a highly genetically related lineage (termed CL1) was dominant among the samples (47 out of 59 samples with good sequencing coverage), spanning the entire period of the collection (2007-2019) and all regions of the country. We also found a second, smaller clonal lineage (termed CL2) of four parasites collected between 2017 and 2019. To explore the regional context of Panamanian P. vivax we conducted principal components analysis and constructed a neighbor-joining tree using these samples and samples collected worldwide from a previous study. Three of the four samples with travel history clustered with samples collected from their suspected country of origin (consistent with importation), while one appears to have been a result of local transmission. The small number of Panamanian P. vivax samples not belonging to either CL1 or CL2 clustered with samples collected from Colombia, suggesting they represent the genetically similar ancestral P. vivax population in Panama or were recently imported from Colombia. The low diversity we observe in Panama indicates that this parasite population has been previously subject to a severe bottleneck and may be eligible for elimination. Additionally, while we confirmed that P. vivax is imported to Panama from diverse geographic locations, the lack of impact from imported cases on the overall parasite population genomic profile suggests that onward transmission from such cases is limited and that imported cases may not presently pose a major barrier to elimination.

Antiviral Activity of Novel Quinoline Derivatives against Dengue Virus Serotype 2.

Dengue virus causes dengue fever, a debilitating disease with an increasing incidence in many tropical and subtropical territories. So far, there are no effective antivirals licensed to treat this virus. Here we describe the synthesis and antiviral activity evaluation of two compounds based on the quinoline scaffold, which has shown potential for the development of molecules with various biological activities. Two of the tested compounds showed dose-dependent inhibition of dengue virus serotype 2 in the low and sub micromolar range. The compounds 1 and 2 were also able to impair the accumulation of the viral envelope glycoprotein in infected cells, while showing no sign of direct virucidal activity and acting possibly through a mechanism involving the early stages of the infection. The results are congruent with previously reported data showing the potential of quinoline derivatives as a promising scaffold for the development of new antivirals against this important virus.

Phage-Displayed Peptides Selected to Bind Envelope Glycoprotein Show Antiviral Activity against Dengue Virus Serotype 2.

Dengue virus is a growing public health threat that affects hundreds of million peoples every year and leave huge economic and social damage. The virus is transmitted by mosquitoes and the incidence of the disease is increasing, among other causes, due to the geographical expansion of the vector's range and the lack of effectiveness in public health interventions in most prevalent countries. So far, no highly effective vaccine or antiviral has been developed for this virus. Here we employed phage display technology to identify peptides able to block the DENV2. A random peptide library presented in M13 phages was screened with recombinant dengue envelope and its fragment domain III. After four rounds of panning, several binding peptides were identified, synthesized, and tested against the virus. Three peptides were able to block the infectivity of the virus while not being toxic to the target cells. Blind docking simulations were done to investigate the possible mode of binding, showing that all peptides appear to bind domain III of the protein and may be mostly stabilized by hydrophobic interactions. These results are relevant to the development of novel therapeutics against this important virus.

Identification of Inhibitors of CD36-Amyloid Beta Binding as Potential Agents for Alzheimer's Disease.

Neuroinflammation is one of the hallmarks of Alzheimer's disease pathology. Amyloid ß has a central role in microglia activation and the subsequent secretion of inflammatory mediators that are associated with neuronal toxicity. The recognition of amyloid ß by microglia depends on the expression of several receptors implicated in the clearance of amyloid and in cell activation. CD36 receptor expressed on microglia interacts with fibrils of amyloid inducing the release of proinflammatory cytokines and amyloid internalization. The interruption of the interaction CD36-amyloid ß compromises the activation of microglia cells. We have developed and validated a new colorimetric assay to identify potential inhibitors of the binding of amyloid ß to CD36. We have found seven molecules, structural analogues of the Trichodermamide family of natural products that interfere with the interaction CD36-amyloid ß. By combining molecular docking and dynamics simulations, we suggested the second fatty acids binding site within the large luminal hydrophobic tunnel, present in the extracellular domain of CD36, as the binding pocket of these compounds. Free energy calculations predicted the nonpolar component as the driving force for the binding of these inhibitors. These molecules also inhibited the production of TNF-α, IL-6, and IL-1ß by peritoneal macrophages stimulated with fibrils of amyloid ß. This work serves as a platform for the identification of new potential anti-inflammatory agents for the treatment of Alzheimer's disease.

Ebony (Phitecellobium flexicaule Benth) and proteins fractionation, solubilization, characterization and production of an isolate.

Different combinations of pHs (2 to 12) and temperatures (25, 30 and 35 degrees C) were tested to obtain a protein isolate from ebony (Pithecellobium flexicaule, Benth) seeds. Seed proteins contained 54.6% albumins, 32% globulins, 5.7% glutelins and 1.3% prolamins. The isoelectric points for albumins, globulins and glutelins were in the pH range of 2.3-2.7. The average molecular weight of albumins ranged from 92 to 100 kDa and for the four globulin subunits in the range of 28.4 to 57.3 kDa. For isolate production, proteins were sequentially extracted with distilled water and a 5% NaCl solution. The resulting supernatants were mixed. The best extraction was achieved at pH 11 and 25 degrees C. 45.6% of the total seed protein was precipitated at pH 2.6 yielding an isolate with 90% protein (N x 6.25). The isolate contained high quantities of lysine, leucine, threonine and phenylalanine but were low in sulfur containing amino acids methionine and cysteine. The extraction process reduced tannins, phytates and trypsin inhibitor in 53, 70 and 70%, respectively. In vivo protein digestibility of the protein isolate was 85.4% and the corrected digestibility essential amino acid score was of 44% due to the lack of sulfur containing amino acids. In order to upgrade the protein quality of ebony isolate it is recommend to supplement with methionine or sulfur containing rich foods.

Análisis descriptivo de los pacientes del departamento 6, varones, del Instituto Psiquiátrico Dr. José Horwitz Barak / Patients' descriptive analysis of departament 6 of men at the Instituto Psiquiátrico Dr. José Horwitz Barak

Se presentan los resultados de un estudio descriptivo realizado por el equipo profesional del Departamento 6 varones de larga estadía del Instituto Psiquiátrico, con la intensión de revisar parámetro, tanto clínicos como sociales y de rehabilitación, para someterlos a un estudio comparativo y confirmar la hipótesis que sugiere que la situación biopsicosocial y de rehabilitación de estos pacientes es significativamente mejor que antes. El estudio incluye resultados obtenidos en 15 pacientes a los cuales se ha incorporado a su esquema terapéutico, el fármaco clozapina. Se demuestran importantes cambios favorables. Se describen además, resultados electroencefalográficos en este grupo. Los resultados corroboran lo planteado en la hipótesis de trabajo. Se verifican avances significativos en su rehabilitación, estado clínico y parámetros sociales

Efecto de la cocción sobre la composición química y valor nutricio de la semilla de Pithecellobium flexicaule (Bent) / Effect of cooking on the chemical and nutritional value of the Pithecellobium flexicaule (Bent) seed

El Pithecellobium flexicaule (Bent) o ébano, leguminosa arbórea de alta productividad distribuida en el Noreste de México, produce semillas tradicionalmente consumidas por comunidades de la región, cocidas tiernas y maduras tostadas. Muestra de tres localidades presentó en las semillas maduras 35.3 por ciento de proteína, 25 por ciento de grasa y 13,2 por ciento de fibra dietaria total (FDT). El tostado por 10 minutos a 80-90 graus Celsius mejora la digestibilidad verdadera de 79.3 a 91.8 por ciento, disminuye hasta 35 por ciento los fitatos y 96 por ciento los inhibidores de tripsina, sin embargo, los taninos como mg equivalentes de catequina/100g se incrementaron de 12.4 a 235.6, ya que éstos pasan durante el tostado de la testa a los cotiledones y la pérdida de aminoácidos azufrados por efecto del tostado ocasiona la disminución del puntaje químico (P.Q.) corregido con digestibilidad verdadera (D.V.) de 44.4 a 34.9. Las semillas tiernas tienen 12.7 por ciento de proteína, 6.6 por ciento de grasa y 3.5 por ciento de FDT. Al cocinarlas tradicionalmente hervidas en su vaina (SVT1), aunque los fitatos se reducen en un 72 por ciento, la digestibilidad es de 85.8 por ciento, igual a la semilla cruda ya que los taninos se incrementaron de 4.9 a 226 mg. Por lo que se probó hervirlas libres de vaina y testa, logrando una reducción de 73.5 por ciento en los taninos, de 88.6 por ciento en los fitatos y digestibilidad de 94.5 por ciento. Igual que en otras leguminosas, los aminoácidos limitantes son los azufrados; el puntaje químico corregido con digestibilidad proteica verdadera de 50.6 por ciento en SVT1, similar al frijol pinto.