Establishing RTS,S/AS01 as a benchmark for comparison to next-generation malaria vaccines in a mouse model.

New strategies are needed to reduce the incidence of malaria, and promising approaches include vaccines targeting the circumsporozoite protein (CSP). To improve upon the malaria vaccine, RTS,S/AS01, it is essential to standardize preclinical assays to measure the potency of next-generation vaccines against this benchmark. We focus on RTS,S/AS01-induced antibody responses and functional activity in conjunction with robust statistical analyses. Transgenic Plasmodium berghei sporozoites containing full-length P. falciparum CSP (tgPb-PfCSP) allow two assessments of efficacy: quantitative reduction in liver infection following intravenous challenge, and sterile protection from mosquito bite challenge. Two or three doses of RTS,S/AS01 were given intramuscularly at 3-week intervals, with challenge 2-weeks after the last vaccination. Minimal inter- and intra-assay variability indicates the reproducibility of the methods. Importantly, the range of this model is suitable for screening more potent vaccines. Levels of induced anti-CSP antibody 2A10 equivalency were also associated with activity: 105 µg/mL (95% CI: 68.8, 141) reduced liver infection by 50%, whereas 285 µg/mL (95% CI: 166, 404) is required for 50% sterile protection from mosquito bite challenge. Additionally, the liver burden model was able to differentiate between protected and non-protected human plasma samples from a controlled human malaria infection study, supporting these models' relevance and predictive capability. Comparison in animal models of CSP-based vaccine candidates to RTS,S/AS01 is now possible under well controlled conditions. Assessment of the quality of induced antibodies, likely a determinant of durability of protection in humans, should be possible using these methods.

Clinical profile of severe hypercholesterolemia in 156,000 adults in primary care. / Perfil clínico de la hipercolesterolemia severa en 156.000 adultos en atención primaria.

OBJECTIVE: To examine the frequency of severe hypercholesterolemia (HS) and its clinical profile, and the phenotype of familial hypercholesterolemia (FH), in the primary-care setting in a large health area of the Community of Madrid (CAM). MATERIAL AND METHODS: Multicenter study of subjects with a health card assigned to 69 health centers (Northwest/CAM area). HS was defined as cholesterol ≥300mg/dL or LDL-cholesterol ≥220mg/dL in any analysis performed (1-1-2018 to 12-30-2021); and FH phenotype as c-LDL ≥240mg/dL (≥160mg/dL if lipid-lowering treatment) with triglycerides <200mg/dL and TSH <5µIU/mL. RESULTS: 156,082 adults ≥18years with an available lipid profile were analyzed. 6187 subjects had HS (3.96% of the laboratory tests studied, 95%CI: 3.87-4.06%). The mean evolution time of the diagnosis of hyperlipidemia in the computerized clinical record was 10.8years, 36.5% had hypertension, 9.5% diabetes and 62.9% overweight/obesity. 83.7% were taking lipid-lowering drugs (65.7% low/moderate and 28.6% high/very high intensity). 6.1% had cardiovascular disease (94.2% treated with lipid-lowering agents), with LDL-cholesterol <55, <70 and <100mg/dL of 1.8%, 5.8% and 20.2%, respectively (vs. 1%, 2.3% and 11.2% if no cardiovascular disease). 1600 subjects had FH phenotype (95%CI: 1.03%, 0.98-1.08%). CONCLUSIONS: Four out of 100 patients analyzed in primary care have HS, with high treatment level, but insufficient intensity, and poor achievement of treatment goals. One in 100 have the FH phenotype. The identification of both dyslipidemias by computerized records would allow their more precise and early detection and establish cardiovascular preventive strategies.

Genistein-mediated thermogenesis and white-to-beige adipocyte differentiation involve transcriptional activation of cAMP response elements in the Ucp1 promoter.

Genistein is an isoflavone present in soybeans and is considered a bioactive compound due to its widely reported biological activity. We have previously shown that intraperitoneal genistein administration and diet supplementation activates the thermogenic program in rats and mice subcutaneous white adipose tissue (scWAT) under multiple environmental cues, including cold exposure and high-fat diet feeding. However, the mechanistic insights of this process were not previously unveiled. Uncoupling protein 1 (UCP1), a mitochondrial membrane polypeptide responsible for dissipating energy into heat, is considered the most relevant thermogenic marker; thus, we aimed to evaluate whether genistein regulates UCP1 transcription. Here we show that genistein administration to thermoneutral-housed mice leads to the appearance of beige adipocyte markers, including a sharp upregulation of UCP1 expression and protein abundance in scWAT. Reporter assays showed an increase in UCP1 promoter activity after genistein stimulation, and in silico analysis revealed the presence of estrogen (ERE) and cAMP (CRE) response elements as putative candidates of genistein activation. Mutation of the CRE but not the ERE reduced genistein-induced promoter activity by 51%. Additionally, in vitro and in vivo ChIP assays demonstrated the binding of CREB to the UCP1 promoter after acute genistein administration. Taken together, these data elucidate the mechanism of genistein-mediated UCP1 induction and confirm its potential applications in managing metabolic disorders.

Engineered DNA-encoded monoclonal antibodies targeting Plasmodium falciparum circumsporozoite protein confer single dose protection in a murine malaria challenge model.

Novel approaches for malaria prophylaxis remain important. Synthetic DNA-encoded monoclonal antibodies (DMAbs) are a promising approach to generate rapid, direct in vivo host-generated mAbs with potential benefits in production simplicity and distribution coupled with genetic engineering. Here, we explore this approach in a malaria challenge model. We engineered germline-reverted DMAbs based on human mAb clones CIS43, 317, and L9 which target a junctional epitope, major repeat, and minor repeat of the Plasmodium falciparum circumsporozoite protein (CSP) respectively. DMAb variants were encoded into a plasmid vector backbone and their expression and binding profiles were characterized. We demonstrate long-term serological expression of DMAb constructs resulting in in vivo efficacy of CIS43 GL and 317 GL in a rigorous mosquito bite mouse challenge model. Additionally, we engineered an Fc modified variant of CIS43 and L9-based DMAbs to ablate binding to C1q to test the impact of complement-dependent Fc function on challenge outcomes. Complement knockout variant DMAbs demonstrated similar protection to that of WT Fc DMAbs supporting the notion that direct binding to the parasite is sufficient for the protection observed. Further investigation of DMAbs for malaria prophylaxis appears of importance.

Evaluation of serological diagnostic tests of human brucellosis for prevention and control in Mexico.

Brucellosis is a zoonosis mainly present in developing countries. The WHO reports 500,000 new cases every year. From 2012 to 2016, 13,677 cases were reported in Mexico, with 2.00 to 2.64 rate per 100,000 inhabitants. To analyze the diagnostic algorithm of brucellosis in Mexico, we compared the commercial laboratory tests ELISA, Brucellacapt®, and lateral flow test (LFT) in a study of 473 individuals from two endemic Mexican populations. All patients were treated in first-level medical units for presenting brucellosis compatible symptoms and without a history of the disease. Clinical-epidemiological information was gathered and initial serum samples were obtained to react with anti-Brucella antibodies; subsequent samples were collected at follow-up treatment visits. Using the Rose Bengal screening, we found 165 negative samples and 308 positive reactive samples, of which 222 cases were confirmed and 234 were positive on at least one marker (IgG or IgM) or LFT. When Brucellacapt® was used, similar results to those observed with the conventional algorithm were found as judged by the Cohen's kappa coefficient (κ) (0.813, 95% CI 0.7788-0.8472). Similar κ indices between conventional algorithm and ELISA pair were found, 0.7038 (95% CI 0.6555-0.7521), representing high similarity between both groups of diagnosis. We conclude that conventional serodiagnoses, Brucellacapt® and LFT, presented inconclusive results and poor correlation between them. By contrast, ELISA test pair (IgG + IgM) presented high correlation with the conventional algorithm and greater capacity for correct positive and negative classification.

TcG2/TcG4 DNA Vaccine Induces Th1 Immunity Against Acute Trypanosoma cruzi Infection: Adjuvant and Antigenic Effects of Heterologous T. rangeli Booster Immunization.

Background: Chagas cardiomyopathy is caused by Trypanosoma cruzi (Tc). Two antigenic candidates, TcG2 and TcG4, are recognized by antibodies in naturally infected dogs and humans; and these vaccine candidates provided protection from Tc infection in mice and dogs. Trypanosoma rangeli (Tr) is non-pathogenic to mammals and shown to elicit cross-reactive anti-Tc antibodies. In this study, we investigated if fixed Tr (fTr) can further enhance the efficacy of the TcG2/TcG4 DNA vaccine. Methods and Results: C57BL/6 mice were immunized with TcG2/TcG4 DNA vaccine and fTr (delivered as an adjuvant or in prime-boost approach), and challenged with Tc. Serology studies showed that fTr (±quil-A) elicited Tc- and Tr-reactive IgGs that otherwise were not stimulated by TcG2/TcG4 vaccine only, and quil-A had suppressive effects on fTr-induced IgGs. After challenge infection, TcG2/TcG4-vaccinated mice exhibited potent expansion of antigen- and Tc-specific IgGs that were not boosted by fTr±quil-A. Flow cytometry analysis showed that TcG2/TcG4-induced dendritic cells (DC) and macrophages (Mφ) responded to challenge infection by expression of markers of antigen uptake, processing, and presentation, and production of pro-inflammatory cytokines. TcG2/TcG4-induced CD4+T cells acquired Th1 phenotype and expressed markers that orchestrate adaptive immunity. A fraction of vaccine-induced CD4+T cells exhibited iTreg phenotype responsible for aversion of self-injurious immune responses. Further, TcG2/TcG4-vaccinated mice exhibited potent expansion of poly-functional CD8+T cells with TNF-α/IFN-γ production and cytolytic phenotype post-infection. Subsequently, tissue parasites and pathology were hardly detectable in TcG2/TcG4-vaccinated/infected mice. Inclusion of fTr±quil-A had no clear additive effects in improving the Tc-specific adaptive immunity and parasite control than was noted in mice vaccinated with TcG2/TcG4 alone. Non-vaccinated mice lacked sufficient activation of Th1 CD4+/CD8+T cells, and exhibited >10-fold higher levels of tissue parasite burden than was noted in vaccinated/infected mice. Conclusion:TcG2/TcG4 vaccine elicits highly effective immunity, and inclusion of fTr is not required to improve the efficacy of DNA vaccine against acute Tc infection in mice.

Paracervical Block for Intrauterine Device Placement Among Nulliparous Women: A Randomized Controlled Trial.

OBJECTIVE: To investigate whether a 20-mL buffered 1% lidocaine paracervical block decreases pain during intrauterine device (IUD) placement. METHODS: In a randomized, single-blind, placebo-controlled trial, women were assigned to receive either a 20-mL buffered 1% lidocaine paracervical block or no block before IUD placement. The primary outcome was pain with IUD placement measured on a 100-mm visual analog scale. Our sample size had 80% power (α=0.05) to detect a 20-mm difference in visual analog scale scores with a SD of 28 mm. Secondary outcomes included pain with speculum placement, paracervical block administration, tenaculum placement, 5 minutes postprocedure, and overall pain perception. RESULTS: From October 7, 2014, through October 26, 2017, 64 women were enrolled and analyzed (33 in the paracervical block arm, 31 in the no-block arm). There were no differences in baseline demographics between the groups. Women who received the paracervical block reported less pain with IUD placement compared with women who received no block (median visual analog scale score of 33 mm vs 54 mm, P=.002). Pain was significantly less in the intervention group for uterine sounding (30 mm vs 47 mm, P=.005), 5 minutes after placement (12 mm vs 27 mm, P=.005), and overall pain perception (30 mm vs 51 mm, P=.015). Participants who received the paracervical block experienced more pain with block administration compared with placebo (30 mm vs 8 mm, P=.003). There was no perceived pain difference for speculum insertion (10 mm vs 6 mm, P=.447) or tenaculum placement (15 mm vs 10 mm, P=.268). CONCLUSION: A 20-mL buffered 1% lidocaine paracervical block decreases pain with IUD placement (primary outcome), uterine sounding (secondary outcome), and 5 minutes after placement (secondary outcome). Although paracervical block administration can be painful, perception of pain for overall IUD placement procedure is lower compared with no block. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, NCT02219308.

A DNA vaccine encoding for TcSSP4 induces protection against acute and chronic infection in experimental Chagas disease.

Immunization of mice with plasmids containing genes of Trypanosoma cruzi induces protective immunity in the murine model of Chagas disease. A cDNA clone that codes for an amastigote-specific surface protein (TcSSP4) was used as a candidate to develop a DNA vaccine. Mice were immunized with the recombinant protein rTcSSP4 and with cDNA for TcSSP4, and challenged with bloodstream trypomastigotes. Immunization with rTcSSP4 protein makes mice more susceptible to trypomastigote infection, with high mortality rates, whereas mice immunized with a eukaryotic expression plasmid containing the TcSSP4 cDNA were able to control the acute phase of infection. Heart tissue of gene-vaccinated animals did not show myocarditis and tissue damage at 365 days following infection, as compared with control animals. INF-γ was detected in sera of DNA vaccinated mice shortly after immunization, suggesting the development of a Th1 response. The TcSSP4 gene is a promising candidate for the development of an anti-T. cruzi DNA vaccine.

Epilepsia en el menor de un año / Epilepsy in children under one year

Se realizó un estudio descriptivo de 92 pacientes, con edad mayor de un mes y menor de un año con diagnóstico definitivo al egreso de epilepsia, pertenecientes al hospital pediátrico provincial de Sancti Spíritus en un período de 5 años (1996-2000). Las variables estudiadas fueron la edad, el sexo, los tipos de crisis que presentaron, la epilepsia o síndrome epiléptico, la etiología y los principales complementarios. Predominó la edad comprendida entre 5 y 8 meses con 54 pacientes para un 58,70 % y el sexo masculino con 58 casos (63,04 %). Los tipos de crisis epilépticas tenemos que fueron más frecuentes las parciales simples motoras con 36 pacientes (39,13 %) y los espasmos infantiles en 17 para un 18,47 %. Las epilepsias y síndromes epilépticos más frecuentes fueron las epilepsias parciales sintomáticas en 29 niños (31,52 %) y el síndrome de West en 17 (18,48 %). En cuanto a la etiología predominaron las epilepsias sintomáticas presentándose en 60 casos para un 65,22 %. Los complementarios más utilizados fueron el electroencefalograma que se le realizó a la totalidad de los casos con un 57,61 % de positividad y la tomografía axial computarizada de cráneo que se les realizó a 86 pacientes con un 67,44 % de positividad.

Tumores cerebrales en el niño. Estudio de cinco años en el Hospital Pediátrico de Sancti Spíritus / Brain tumors in children. A five year study in the Pediatric Hospital of Sancti Spíritus

Los tumores primarios del sistema nervioso central, constituyen el segundo tipo de cáncer infantil, siendo superados solamente por leucemias. Con el objetivo de descubrir un grupo de características clínico epidemiológico y de laboratorio, se realizó un estudio descriptivo retrospectivo de 13 niños pertenecientes al Hospital Pediátrico Docente Provincial de Sancti Spíritus con diagnóstico de tumor cerebral primario en el período comprendido entre 1995 a 1999. La edad más frecuente de presentación de los mismos fue la comprendida entre los 6 a 10 años con más de la mitad de los casos. Predominó el sexo masculino sobre el femenino. Los síntomas iniciales fundamentales fueron la cefalea, los trastornos de la marcha, y las náuseas y/o vómitos, en tanto que los signos fueron el papiledema, la ataxia y la paresia de nervios oculomotores. Alrededor del 70% de los tumores eran de localización infratentorial. Los tipos histológicos más frecuentes fueron el meduloblastoma de fosa posterior, el astrocitoma del cerebelo y el craneofaringioma. LaTomografía axial computarizada fue positiva en el 92,3% de los casos.

Parasomnia de la infancia / Childhood parasomnia

Este trabajo describe un grupo de características en niños egresados de la sala de neurología que presentaron parasomnias como fueron: la distribución según edad y sexo, señalando el motivo principal de ingreso en cada caso, identificando el tipo de parasomnia que presentaron y clasificando la parasomnias según el grado de las mismas.

La variante letal del síndrome Prune Belly: informe de dos casos / The lethal variant of the \"Prune Belly's\" syndrome: two cases report

En este informe se describen dos casos de la variante letal del síndrome de Prune Belly en que la obstrucción de la uretra produjo graves alteraciones del sistema urinario con distensión vesical y ureteral con repercusión secundaria en la pared abdominal y daño renal bilateral por displasia. Se comenta la etiopatogenia de este síndrome de acuerdo a la teoría embriológica del mesodermo, así como de la teoría de la secuencia de la obstrucción uretral en las etapas tempranas del desarrollo que repercuten en la pared abdominal. En estos casos descritos, el grado de obstrucción del tracto urinario repercutió en el desarrollo de malformaciones letales

Estudio de 60 pacientes pediátricos con convulsiones febriles / Study of 60 pediatric patients with febrile convulsions

Las crisis febriles son muy frecuentes en el paciente pediátrico afectando entre el 3 - 5 % de los niños entre los tres meses y los cinco años de edad. Se realizó un estudio descriptivo retrospectivo de 60 pacientes con el diagnóstico al egreso de crisis febriles pertenecientes al Hospital Pediátrico Docente de Sancti Spíritus en un período de un año con el objetivo de describir un grupo de características clínicas y electroencefalográficas. La edad de debut más frecuente fue la de 13 - 24 meses (58,34 %), la mayoría de los pacientes tenían temperatura entre 38 - 39,5 grados Celsius (60 %). Las crisis se presentaron en el 65 % de los casos en las primeras seis horas de la fiebre, la causa más común fueron las infecciones respiratorias altas (48,33 %) según las características de las crisis el 98,33 % fueron generalizadas, el 93,33 % duró menos de 15 minutos y el 95 % se presentaron en las primeras 24 horas de la fiebre. El 86,67 % de las crisis fueron simples y el 13,33 % complejas. Solamente el 36,67 % de los casos tuvo recurrencias, el EEG fue normal en el 93,33 % de los casos.