A redox-active crosslinker reveals an essential and inhibitable oxidative folding network in the endoplasmic reticulum of malaria parasites.

Malaria remains a major global health problem, creating a constant need for research to identify druggable weaknesses in P. falciparum biology. As important components of cellular redox biology, members of the Thioredoxin (Trx) superfamily of proteins have received interest as potential drug targets in Apicomplexans. However, the function and essentiality of endoplasmic reticulum (ER)-localized Trx-domain proteins within P. falciparum has not been investigated. We generated conditional mutants of the protein PfJ2-an ER chaperone and member of the Trx superfamily-and show that it is essential for asexual parasite survival. Using a crosslinker specific for redox-active cysteines, we identified PfJ2 substrates as PfPDI8 and PfPDI11, both members of the Trx superfamily as well, which suggests a redox-regulatory role for PfJ2. Knockdown of these PDIs in PfJ2 conditional mutants show that PfPDI11 may not be essential. However, PfPDI8 is required for asexual growth and our data suggest it may work in a complex with PfJ2 and other ER chaperones. Finally, we show that the redox interactions between these Trx-domain proteins in the parasite ER and their substrates are sensitive to small molecule inhibition. Together these data build a model for how Trx-domain proteins in the P. falciparum ER work together to assist protein folding and demonstrate the suitability of ER-localized Trx-domain proteins for antimalarial drug development.

An Endoplasmic Reticulum CREC Family Protein Regulates the Egress Proteolytic Cascade in Malaria Parasites.

The endoplasmic reticulum (ER) is thought to play an essential role during egress of malaria parasites because the ER is assumed to be required for biogenesis and secretion of egress-related organelles. However, no proteins localized to the parasite ER have been shown to play a role in egress of malaria parasites. In this study, we generated conditional mutants of the Plasmodium falciparumendoplasmic reticulum-resident calcium-binding protein (PfERC), a member of the CREC family. Knockdown of the PfERC gene showed that this gene is essential for asexual growth of P. falciparum Analysis of the intraerythrocytic life cycle revealed that PfERC is essential for parasite egress but is not required for protein trafficking or calcium storage. We found that PfERC knockdown prevents the rupture of the parasitophorous vacuole membrane. This is because PfERC knockdown inhibited the proteolytic maturation of the subtilisin-like serine protease SUB1. Using double mutant parasites, we showed that PfERC is required for the proteolytic maturation of the essential aspartic protease plasmepsin X, which is required for SUB1 cleavage. Further, we showed that processing of substrates downstream of the proteolytic cascade is inhibited by PfERC knockdown. Thus, these data establish that the ER-resident CREC family protein PfERC is a key early regulator of the egress proteolytic cascade of malaria parasites.IMPORTANCE The divergent eukaryotic parasites that cause malaria grow and divide within a vacuole inside a host cell, which they have to break open once they finish cell division. The egress of daughter parasites requires the activation of a proteolytic cascade, and a subtilisin-like protease initiates a proteolytic cascade to break down the membranes blocking egress. It is assumed that the parasite endoplasmic reticulum plays a role in this process, but the proteins in this organelle required for egress remain unknown. We have identified an early ER-resident regulator essential for the maturation of the recently discovered aspartic protease in the egress proteolytic cascade, plasmepsin X, which is required for maturation of the subtilisin-like protease. Conditional loss of PfERC results in the formation of immature and inactive egress proteases that are unable to breakdown the vacuolar membrane barring release of daughter parasites.

Role of progesterone receptor isoforms in female sexual behavior induced by progestins in rats.

Progesterone and its ring A reduced metabolites regulate female sexual behavior through the direct or indirect activation of progesterone receptor (PR) which has two isoforms with different function and regulation: PR-A and PR-B. The contribution of each PR isoform to the regulation of lordosis in rats is unknown. We explored the role of PR isoforms in lordosis display induced by progesterone and two of its ring A reduced metabolites: 5alpha-pregnan-3,20-dione (5alpha-DHP), and 5beta,3beta-pregnan-20-one (5beta,3beta-Pgl) in adult ovariectomized rats. Two weeks after ovariectomy, the animals were injected subcutaneously with 5 microg of estradiol benzoate (EB), and 40 h later, progestins were injected intracerebroventricularly. PR-B and total PR (PR-A + PR-B) sense or antisense oligonucleotides were administered intracerebroventricularly immediately before EB injection and 24 h later. Lordosis was evaluated 30, 120 and 240 min after progestin administration. Western blot analysis of both PR isoforms was performed in the hypothalamus and preoptic area 24 h after lordosis tests. All progestins induced maximal lordosis 120 min after administration, and antisense oligonucleotides against both PR isoforms inhibited lordosis in all animals. PR-B antisense oligonucleotides also inhibited lordosis induced by progesterone and 5alpha-DHP although with less efficacy than total PR antisense oligonucleotides, but the former inhibited lordosis induced by 5beta,3beta-Pgl in a similar manner as total PR antisense oligonucleotides. In the hypothalamus and preoptic area, the content of both PR isoforms or PR-B alone was diminished by the administration of total or PR-B antisense oligonucleotides, respectively. These results suggest that the PR-B isoform is essential for the display of the lordosis behavior in rats.

Síndromes geriátricos y abandono

Fundamento: el abandono en la vejez es un condicionante de muchos trastornos de salud y constituye en mayor medida una condición de gran incidencia a nivel mundial. Objetivo: comparar los síndromes geriátricos y el abandono en pacientes adultos mayores. Métodos: se realizó un estudio transversal, descriptivo, analítico y correlacional, en la Unidad de Medicina Familiar No. 33 del Instituto Mexicano del Seguro Social de Tabasco. La muestra se conformó con 203 pacientes de 60 años y más, que acudieron a consulta de Medicina Familiar y Atención Médica Continua. Se aplicaron 3 instrumentos: la escala de Pfeiffer, el International Consultation on Incontinence Questionnaire Short-Form, la Escala de Percepción de Abandono del Adulto Mayor y se interrogó sobre el consumo de medicamentos prescritos y automedicados. Resultados: existió relación entre los síndromes geriátricos y el abandono, como se comprobó en la correlación entre las variables analizadas. El deterioro cognitivo leve, moderado y severo, se encontró en bajas proporciones. El 40,7 % de los pacientes aceptó tener incontinencia urinaria. La presencia de polifarmacia se encontró en altas proporciones. Conclusiones: los síndromes geriátricos presentes en el estudio fueron: deterioro cognitivo, polifarmacia e incontinencia urinaria, más la presencia de ser soltero, viudo o divorciado muestran una relación significativa para sufrir abandono.

Foundation: abandonment in old age is a condition of many health disorders and is, to a greater extent, a condition of high incidence worldwide. Objective: to compare geriatric syndromes and abandonment in older adult patients. Methods: a cross-sectional, descriptive, analytical and correlational study was carried out in the Family Medicine Unit No. 33 of the Mexican Social Security Institute of Tabasco. The sample was made up of 203 patients aged 60 and over, who attended Family Medicine and Continuing Medical Care consultations. Three instruments were applied: the Pfeiffer scale, the International Consultation on Incontinence Questionnaire Short-Form, the Perception of Abandonment Scale for the Elderly, and questions were asked about the consumption of prescribed and self-medicated medications. Results: there is a relationship between geriatric syndromes and abandonment, as verified in the correlation between the variables analyzed. Mild, moderate and severe cognitive impairment was found in low proportions. 40.7 % of the patients accepted having urinary incontinence. The presence of polypharmacy was found in high proportions. Conclusions: the geriatric syndromes present in the study such as: cognitive impairment, polypharmacy and urinary incontinence, plus the presence of being single, widowed or divorced show a significant relationship to suffering abandonment.

Diagnosis and outcomes of pregnant women with Zika virus infection in two municipalities of Risaralda, Colombia: Second report of the ZIKERNCOL study.

BACKGROUND: Zika virus (ZIKV) infection has emerged as a significant threat for pregnant women and newborns in populations living in or visiting Latin America. We previously reported a preliminary analysis in Sucre, Colombia, as the first group of pregnant women with RT-PCR-confirmed ZIKV (ZIKa enEmbarazadas yReciénNacidos enCOLombia, ZIKERNCOL). METHODS: In this second report, findings of the first 86 pregnant women from La Virginia and Dosquebradas (municipalities), Risaralda, Colombia, with RT-PCR-confirmed ZIKV infection are reported. Clinical, demographical and obstetrical findings are described. RESULTS: All women reported ZIKV symptoms during pregnancy: 79.1% rash, 55.8% fever, among others. In addition to ZIKV, RT-PCR was positive for dengue in 18.6%; 45.3% Dengue IgM+; 5.8% RT-PCR positive for chikungunya; 3.6% Chikungunya IgM+. STORCH screening in mother: 11.6% IgG + anti-Toxoplasma gondii, 6% IgG + anti-rubella, 4.7% IgG + CMV. The rest of STORCH tests were negative. Microcephaly was observed in 2.4% of the newborns. No calcifications or other CNS alterations were detected. One newborn had cleft palate and one had bilateral renal ectopy. CONCLUSIONS: The rate of microcephaly in our cohort was consistent with other studies. Pregnant women in endemic areas should be followed and tested according to standard protocols, and asymptomatic ZIKV infection should be considered. Long-term follow-up of children is required in the congenital Zika syndrome (CZS) assessment.

Zika virus: clinical manifestations and treatment at a primary care institution in Colombia / Virus del Zika: manifestaciones clínicas y tratamiento en una institución de primer nivel en Colombia

Abstract Introduction: Although Zika virus cases have been reported in Colombia since 2015, its clinical and pharmacological characteristics have not yet been described. Objective: To describe the main clinical manifestations and sociodemographic characteristics of patients diagnosed with Zika and the treatment provided to them at a primary care hospital in a municipality of Colombia. Materials and methods: Descriptive cross-sectional study. The study population consisted of patients diagnosed with Zika between January 1 and July 25, 2016 at a primary care hospital. Sociodemographic, clinical, and pharmacological variables, as well as adverse clinical outcomes associated with the infection were included. Descriptive statistics were performed. A x2 test was used for categorical variables, and a multivariate analysis was conducted using Epi info 7.1 software. Results: 254 individuals infected with Zika virus during the study period were identified, and Zika diagnosis was more frequent in women (68.5%). Regarding treatment, 90.9% of the cases were treated using acetaminophen. The most commonly reported symptoms were rash (81.1%) and pruritus (55.9%). In addition, antihistamines were the most frequent comedication (31.9%). Factors such as being a woman, being pregnant and inpatient treatment were associated with adverse clinical outcomes. Conclusion: The clinical manifestations described here are similar to those reported in other populations. Furthermore, inappropriate pharmacological management practices that can lead to complications in this population, such as bleeding, were observed in some cases. Thus, educational interventions on the proper prescription of medications for treating this disease aimed at general physicians working in Zika affected areas must be implemented to improve the prognosis of these patients.

Resumen Introducción. Aunque el virus del Zika está presente en Colombia desde el 2015, sus características clínicas y farmacológicas aún no han sido descritas. Objetivo. Describir las principales manifestaciones clínicas y características sociodemográficas de pacientes diagnosticados con zika, así como el tratamiento que recibieron en un hospital de primer nivel. Materiales y métodos. Estudio descriptivo de corte transversal. La población estuvo constituida por los pacientes diagnosticados con zika entre el 1 de enero y el 25 de julio de 2016 en un hospital de primer nivel. Se incluyeron variables sociodemográficas, clínicas y farmacológicas, además de los resultados adversos clínicos y paraclínicos asociados a la infección. Se realizó estadística descriptiva; para las variables categóricas se usó la prueba x2 y el análisis multivariado se realizó a través del programa Epi Info 7.0. Resultados. Se identificaron 254 pacientes con zika, siendo más frecuente en mujeres (68.5%). El 90.9% de la población recibió tratamiento con acetaminofén. Los síntomas más comunes fueron sarpullido (81.1%) y prurito (55.9%). Además, los antihistamínicos fueron la comedicación más frecuente (31.9%). Los factores asociados con resultados clínicos y paraclínicos adversos fueron ser mujer, estar embarazada y tener manejo intrahospitalario. Conclusión. Las manifestaciones clínicas encontradas fueron similares a las reportadas en otras poblaciones. En algunos casos se observó un manejo farmacológico no recomendado, lo que puede generar complicaciones como sangrados; en consecuencia, se deben implementar intervenciones educativas sobre la prescripción adecuada de medicamentos para tratar esta enfermedad, dirigidas a médicos generales que trabajen en regiones afectadas por el zika para, así, mejorar el pronóstico de estos pacientes.

Progesterone receptor isoforms differentially regulate the expression of tryptophan and tyrosine hydroxylase and glutamic acid decarboxylase in the rat hypothalamus.

Progesterone exerts a variety of actions in the brain through the interaction with its receptors (PR) which have two isoforms with different function and regulation: PR-A and PR-B. Progesterone may modulate neurotransmission by regulating the expression of neurotransmitters synthesizing enzymes or their receptors in several brain regions. The role of PR isoforms in this modulation is unknown. We explored the role of PR isoforms in the regulation of tryptophan (TPH) and tyrosine (TH) hydroxylase, and glutamic acid decarboxylase (GAD) expression in the hypothalamus of ovariectomized rats. Two weeks after ovariectomy, animals were subcutaneously injected with 5 µg of estradiol benzoate (EB), and 40 h later, progesterone (P) was intracerebroventricularly (ICV) injected. Each animal received two ICV injections of 1 µg/µl (4 nmol) of PR-B and total PR (PR-A+PR-B) sense or antisense (As) oligonucleotides (ODNs). First injection was made immediately before sc EB injection, and 24h later animals received the second one. Twenty-four hours after P administration, rats were euthanized and brains removed to measure the expression of PR-A and PR-B, TPH, TH and GAD by Western blot. We observed that sense ODNs modified neither PR isoforms nor enzymes expression in the hypothalamus, whereas PR A+B antisense (PR A+B As) clearly decreased the expression of both PR isoforms in this region. ICV administration of PR-B As only decreased PR-B isoform expression with no significant effects on PR-A expression. A differential protein expression of TPH, TH and GAD was observed after PR isoforms antisense administration. PR-B As administration decreased the expression of TPH (65% with respect to control). In contrast, PR A+B As and PR-B As administration increased (51.6% and 34.4%, respectively) TH expression. The administration of PR A+B As and PR-B As diminished GAD expression (33.4% and 41.6%, respectively). Our findings indicate that PR isoforms play a differential role in the regulation of the content of TPH, TH and GAD in the rat hypothalamus.

Anticoagulación oral en pacientes con Insuficiencia Cardiaca Crónica con fracción de eyección reducida, experiencia en el centro Médico Quirúrgico Boliviano-Belga / Oral anticoagulation on patients with chronic hearth failure with reduced ejection fraction, experience at Centro Médico Quirúrgico Boliviano Belga

Objetivos: describir la experiencia de la anticoagulación oral con warfarina en pacientes con insuficiencia cardiaca crónica con fracción de eyección disminuida (ICC-FER) y la incidencia de ictus y mortalidad en el Centro Médico Quirúrgico Boliviano Belga durante los años 2004 - 2011. Métodos: estudio descriptivo, transversal en >18 años con ICC-FER sintomática, dividiéndolos en los que presentan ritmo sinusal y fibrilación auricular no valvular (FANV), excluyendo quienes tienen trombos intracavitarios, tromboembolia reciente, prótesis valvular y los que requirieron su implante durante el estudio. Resultados: 46 pacientes con ritmo sinusal y 18 con FANV, con fracción de eyección severamente reducida en ambos grupos (29% y 30% respectivamente), todos con tratamiento diurético y modificadores de enfermedad. Todos los pacientes con FANV recibieron tratamiento antitrombótico (anticoagulantes y/o antiagregantes plaquetarios), el 94% con indicación de anticoagulación según CHA2DS2 VASc, estaban medicados con warfarina. El 67% de los pacientes con ritmo sinusal fue anticoagulado y el 6,5% no. La incidencia de ictus fue de 0,11 en FANV y 0,15 en ritmo sinusal en 3 años de seguimiento, mientras que la mortalidad alcanzó el 40% y el 42% respectivamente en un periodo promedio de 4 años (±1,2). Conclusiones: en ICC-FER, se anticoagula a la mayoría de pacientes con FANV, que en este grupo, el riesgo embólico alto. La anticoagulación oral con warfarina en insuficiencia cardiaca crónica con FE severamente comprometida (< 30%) en ritmo sinusal es frecuente, sin reportar complicaciones mayores. La incidencia de ictus fue baja y la mortalidad global fue elevada.

Objectives: to describe warfarin use and the incidence of stroke and mortality in chronic hearth failure with reduced ejection fraction (HFrEF) at Centro Médico Quirúrgico Boliviano Belga between 2004 and 2011. Methods: descriptive and transversal study in > 18 years with symptomatic ICC-FER, dividing them into those with sinus rhythm and non-valvular atrial fibrillation (NVAF), excluding those with intracavitary thrombus, recent thromboembolism, prosthetic valve and requiring the implant during the study. Results: 46 patients with sinus rhythm and 18 with NVAF. 46 patients with sinus rhythm and 18 with NVAF with severely reduced ejection fraction in both groups (29% and 30% respectively), all with diuretics and disease modifiers. All NVAF patients received antithrombotic therapy (anticoagulant and/or antiplatelet agents), 94% indicating anticoagulation as VASc CHA2DS2 were medicated with warfarin.). All the patients with NVAF received antithrombotics (anticoagulants and/or and antiplatelet drugs), 94% of which required anticoagulation according to CHA2DS2 VASc, they were medicated with warfarin. 67% of patients with sinus rhythm were anticoagulated whilst 6,5% were not. The stroke incidence were of 0,11 in NVAF and 0,15 in sinus rhythm during a 3 year monitoring period, whilst mortality reached 40% and 42% respectively in an average period of 4 years (+1,2). Conclusions: in HFrEF and NVFA most patients where receiving warfarin, that in this group had high embolic risk. Oral anticoagulation with warfarin in chronic heart failure with severely compromised FE (< 30%) in sinus rhythm often, without reporting major complications. The incidence of stroke was low and the overall mortality was high.

Relación entre rritropoyetina profiláctica en aemia del prematuro de muy bajo peso y frecuencia transfusional, hospital materno infantil germán urquidi, marzo 2009 a enero 2010 / Relationship between prophylactic Erythropoietin on Anemia of prematurity in very low weights births and often transfusión, "Germán Urquidi" Maternoinfantil Hospital, March 2009 to January 2010

La anemia es una patología frecuente en el prematuro de muy bajo peso requiriendo reiteradas transfusiones sanguíneas, las cuales presentan numerosos riesgos. La Eritropoyetina influye en el desarrollo y aumento de la población de glóbulos rojos. Determinar si la administración de Eritropoyetina profiláctica en la anemia del prematuro de muy bajo peso disminuye la frecuencia transfusional. Estudio retrospectivo de cohorte interna de 24 prematuros de muy bajo peso con edad gestacional < 32 semanas: un grupo expuesto a Eritropoyetina recombinante humana y el otro no. Se excluyen defunciones, anemia previa, sepsis, retardo de crecimiento intrauterino, insuficiencia cardiaca congestiva, cirugías o malformaciones congénitas mayores. Grupo expuesto: peso 1000 g., media de 2 trasfusiones, 54 días de internación, sin anemia al alta; grupo sin exposición: 1250 g., 2,23 transfusiones y 38 días de hospitalización. La utilización de Eritropoyetina no redujo la frecuencia transfusional en el tratamiento de la anemia del prematuro de muy bajo peso, ni evitó su producción.

Very low weight preterm newborn have commonly anemia, requiring repeated blood transfusions, of which are many risks. Erythropoietin influences development and increases population of red blood cells. Determine if the use of Erythropoietin to prevent Anemia of prematurity reduces red-cell transfusions frequency. Internal retrospective cohort study. 24 low weight preterm newborn < 32 gestational weeks age. Excluding deaths, prior anemia, intraventricular hemorrhage, sepsis, delay of intrauterine growth, congestive heart failure, surgeries or major congenital malformations. Exposed group: Weight 1000 g., average of 2 transfusions and 54 days of placement; not exposed: 1250 g., 2.23 transfusions and 38 days hospitalized. The use of Erythropoietin can't reduces transfusions in Anemia 's very low weight preterm newborn treatment, neither prevent its production.