Intestinal polyparasitism and levels of mucosal anthelmintic SIgA in children from endemic areas in Northeastern Brazil.

Interactions between parasites during co-infections are often complex and can impact immunization and treatment programmes, as well as disease outcomes and morbidity. However, little is known about these interactions and the mechanisms involved. In this study, a coproparasitological survey was carried out in school-age children living in endemic areas of parasitic infection in the state of Sergipe, Northeastern Brazil. Anti-helminth-specific and total secretory immunoglobulin-A (SIgA) levels were measured in stool and saliva samples and were compared in children presenting monoparasitism, polyparasitism (helminths and/or intestinal protozoa) and no infections. The survey showed that protozoa were more prevalent than helminths, and that there was a high frequency of polyparasitism in the studied population, mainly from combinations of protozoan species. Although less frequent, combinations between species of protozoa and helminths were also observed. The levels of salivary SIgA in these co-infected individuals were lower than the average observed in infections with helminths alone. Although the children participating in this survey were asymptomatic, and it was, therefore, not possible to evaluate the impact of salivary SIgA reduction on the diseases, and the study highlights the need for further investigations of co-infections by intestinal parasites and the effects on immune response induced by the interactions between different parasites.

Changes in the epidemiological profile of intestinal parasites after a school-based large-scale treatment for soil-transmitted helminths in a community in northeastern Brazil: Epidemiological profile after large-scale school-based treatment for STH.

Intestinal parasites cause a significant public health problem worldwide due to the associated morbidities, mainly in infected school-aged children (SAC). The strategy of large-scale deworming in SAC to control the transmission of soil-transmitted helminths (STH) has been advocated by the World Health Organization and was recently adopted in Brazil; however, the long-term effects of mass deworming on the larger parasitological profile have been less studied. After a five-year period of school-based large-scale treatment for STH using an annual single dose of albendazole in a community of Sergipe state, Brazil, a marked reduction in prevalence was observed (15.4%% vs.7.4% for Ascaris sp., 6.0%%  vs. 0.4% for hookworm, and 12.8%%  vs. 4.5%% for Trichuris trichiura), with the exception of Strongyloides stercoralis, which had no statistically significant change in prevalence. There was, however, an increase in the prevalence of intestinal protozoans, specifically Entamoeba histolytica/E. dispar (0.0%% vs. 36.0%), Blastocystis hominis (0.0%%  vs. 40.1%), and Giardia duodenalis (5.6%% vs. 14.5%). Although the findings showed a dramatic reduction in the prevalence of STH after four rounds of preventive chemotherapy, there was an increase in intestinal protozoan infections, indicating a change in the epidemiological profile.

Protocolo de Controle de tronco em Ambiente Aquático para Crianças com Paralisia Cerebral: Ensaio Clínico Randomizado / Protocol of Control of Trunk in Aquatic Environment for Children with Cerebral Palsy: Randomized Clinical Trial

Objetivos: Avaliar os efeitos de um protocolo de controle de tronco em ambiente aquático e sua repercussão na funcionalidade de indivíduos com paralisia cerebral (PC) diparéticoespástico, classificados no nível IV do GMFCS (Gross Motor Function Classification System). Métodos: ensaio clínico controlado, randomizado, cego, descritivo-analítico, quantitativo. Foram triados 92 prontuários, 24 crianças foram incluídas e 22 finalizaram o estudo. Os pacientes foram alocados em grupo controle (GC), que realizou terapias convencionais e grupo intervenção (GI) que realizou o protocolo de exercícios aquáticos. Os grupos foram avaliados pré e pós-intervenção através da Trunk Control Measurement Scale (TCMS), Pediatric Reach Test (PRT), Eletromiografia de Superfície (EMG) dos músculos reto abdominal e latíssimo do dorso. Para análises estatísticas foram utilizados o teste de Kolmogorov-Smirnov no momento pré e pós intervenção, teste de Mann-Whitney para a análise intragrupo e teste de Wilcoxon para análise intergrupo. A Correlação de Spearman foi utilizada para observar o grau de associação entre duas variáveis. Foi considerado um intervalo de confiança (IC) de 95%, nível de significância de p<0,05. Resultados: na análise intragrupo constatou-se melhora no item reação de equilíbrio da TCMS, GI (p=0,019) e o GC (p=0,004). No PRT, GC apresentou maior deslocamento pós-intervenção (p=0,006) que o GI. No item 07 da TCMS houve melhora da ativação muscular do reto abdominal (RA) do GI (p=0,047). Conclusão: No presente estudo, observou-se que a fisioterapia aquática trouxe resultados positivos e ganhos motores relacionados ao controle de tronco e funcionalidade para crianças com paralisia cerebral diparética espásticas GMFCS nível IV. (AU)

Objectives: To evaluate the effects of a trunk control protocol in the aquatic environment and its impact on the functionality of individuals with spastic diparesis type of CP (Cerebral Palsy) classified as level IV on GMFCS. Methods: A quantitative, controlled clinical trial, randomized, blind, of descriptive and analytical character. 92 records were screened, 24 children were enrolled and 22 completed the study. Patients were allocated in control group (CG), which held conventional therapies and intervention group (IG) which performed the aquatic exercise protocol. Groups were evaluated pre and post intervention through Trunk Control Measurement Scale (TCMS), Pediatric Reach Test (PRT), Surface Electromyography (EMG) of rectus abdominis and latissimus dorsi muscles. For statistical analysis, it was used the Kolmogorov-Smirnov test for pre and post intervention analysis, Mann-Whitney and Wilcoxon test for intragroup and intergroup analysis, respectively. Spearman correlation was used to observe the rate of association between two variables. A confidence interval (CI) of 95% was considered, level of significance of p <0.05. Results: Intragroup analysis showed improvement in TCMS equilibrium reactions item in GI (p = 0.019) and CG (p = 0.004). In the PRT, GC presented greater post-intervention displacement (p = 0.006) than the GI. In item 07 of the TCMS there was improvement of rectus abdominis muscle activation in GI (p = 0.047). Conclusion: In the present study there were positive results and motor gains on trunk control and functionality for individuals with spastic diparesis type of CP classified as level IV on GMFCS after aquatic therapy intervention. (AU)

Gender Identity and Sexual Function in 46,XX Patients with Congenital Adrenal Hyperplasia Raised as Males.

In individuals with congenital adrenal hyperplasia (CAH) and 46,XX karyotype, androgens produced by the adrenal glands during the intrauterine development promote virilization of the genitals, which may even result in the development of a well-formed penis. Some of these children with late diagnosis are registered as males after birth. After obtaining approval from the internal review board, we evaluated gender identity and sexual function in four 46,XX severely virilized patients with CAH, who were originally registered and raised as males, assisted in our Disorders of Sexual Development Clinic. The evaluation consisted of questionnaires to assess gender identity and sexual activity and interview with the multidisciplinary team that provides care for these patients. The patients underwent surgery to remove uterus, ovaries, and remaining vaginal structures, in addition to implantation of testicular prosthesis and correction of hypospadias, when necessary. All four patients have developed a clear male gender identity, and when evaluated for sexual activity, they have reported having erections, libido, orgasms, and sexual attraction to women only. Two of these 4 patients had satisfactory sexual intercourses when assessed using the International Index of Erectile Function questionnaire. The other two patients who never had sexual intercourse reported not having a partner for sexual activity; one is 18 years old, and the other is 14 years old. This study showed that this group of 46,XX severely virilized patients with CAH, registered and raised as males, adapted well to the assigned male gender, with satisfactory sexual function in patients who had sexual intercourse.

Modeling mutant/wild-type interactions to ascertain pathogenicity of PROKR2 missense variants in patients with isolated GnRH deficiency.

A major challenge in human genetics is the validation of pathogenicity of heterozygous missense variants. This problem is well-illustrated by PROKR2 variants associated with Isolated GnRH Deficiency (IGD). Homozygous, loss of function variants in PROKR2 was initially implicated in autosomal recessive IGD; however, most IGD-associated PROKR2 variants are heterozygous. Moreover, while IGD patient cohorts are enriched for PROKR2 missense variants similar rare variants are also found in normal individuals. To elucidate the pathogenic mechanisms distinguishing IGD-associated PROKR2 variants from rare variants in controls, we assessed 59 variants using three approaches: (i) in silico prediction, (ii) traditional in vitro functional assays across three signaling pathways with mutant-alone transfections, and (iii) modified in vitro assays with mutant and wild-type expression constructs co-transfected to model in vivo heterozygosity. We found that neither in silico analyses nor traditional in vitro assessments of mutants transfected alone could distinguish IGD variants from control variants. However, in vitro co-transfections revealed that 15/34 IGD variants caused loss-of-function (LoF), including 3 novel dominant-negatives, while only 4/25 control variants caused LoF. Surprisingly, 19 IGD-associated variants were benign or exhibited LoF that could be rescued by WT co-transfection. Overall, variants that were LoF in ≥ 2 signaling assays under co-transfection conditions were more likely to be disease-associated than benign or 'rescuable' variants. Our findings suggest that in vitro modeling of WT/Mutant interactions increases the resolution for identifying causal variants, uncovers novel dominant negative mutations, and provides new insights into the pathogenic mechanisms underlying heterozygous PROKR2 variants.

Molecular cloning, biochemical characterization, and partial protective immunity of the heme-binding glutathione S-transferases from the human hookworm Necator americanus.

Hookworm glutathione S-transferases (GSTs) are critical for parasite blood feeding and survival and represent potential targets for vaccination. Three cDNAs, each encoding a full-length GST protein from the human hookworm Necator americanus (and designated Na-GST-1, Na-GST-2, and Na-GST-3, respectively) were isolated from cDNA based on their sequence similarity to Ac-GST-1, a GST from the dog hookworm Ancylostoma caninum. The open reading frames of the three N. americanus GSTs each contain 206 amino acids with 51% to 69% sequence identity between each other and Ac-GST-1. Sequence alignment with GSTs from other organisms shows that the three Na-GSTs belong to a nematode-specific nu-class GST family. All three Na-GSTs, when expressed in Pichia pastoris, exhibited low lipid peroxidase and glutathione-conjugating enzymatic activities but high heme-binding capacities, and they may be involved in the detoxification and/or transport of heme. In two separate vaccine trials, recombinant Na-GST-1 formulated with Alhydrogel elicited 32 and 39% reductions in adult hookworm burdens (P < 0.05) following N. americanus larval challenge relative to the results for a group immunized with Alhydrogel alone. In contrast, no protection was observed in vaccine trials with Na-GST-2 or Na-GST-3. On the basis of these and other preclinical data, Na-GST-1 is under possible consideration for further vaccine development.

Fructose-1,6-bisphosphate inhibits in vitro and ex vivo platelet aggregation induced by ADP and ameliorates coagulation alterations in experimental sepsis in rats.

Sepsis is a systemic response to an infection that leads to a generalized inflammatory reaction. There is an intimate relationship between procoagulant and proinflammatory activities, and coagulation abnormalities are common in septic patients. Pharmaceutical studies have focused to the development of substances that act on coagulation abnormalities and on the link between coagulation and inflammation. Fructose-1,6-bisphosphate (FBP) is a high-energy glycolitic metabolite that in the past two decades has been shown therapeutic effects in great number of pathological situations, including sepsis. The aims of this study were to assess the effects of FBP on platelet aggregation in vitro and ex vivo in healthy and septic rats and evaluate the use of FBP as a treatment for thrombocytopenia and coagulation abnormalities in abdominal sepsis in rat. FBP inhibited platelet aggregation (P < 0.001) in vitro in healthy rats from the smallest dose tested, 2.5 mM, in a dose-dependent manner. The mean effective dose calculated was 10.6 mM. The highest dose tested, 40 mM, completely inhibited platelet aggregation (P < 0.001) induced by ADP. Platelet aggregation in plasma from septic rats was inhibited only with higher doses of FBP, starting from 20 mM (P < 0.001). The calculated mean effective dose was 19.3 mM. Ex vivo platelet aggregation in septic rats was significantly lower (P < 0.05) than healthy rats and the treatment with FBP, at the dose of 2 g/kg, diminished the platelet aggregation at the extension of 27% (P < 0.001), suggesting that FBP is a potent platelet aggregation inhibitor in vivo. Moreover, treatment with FBP 2 g/kg prevented thrombocytopenia (P < 0.001), prolongation of prothrombin and partial thromboplastin time (P < 0.001), but not fibrinogen, in septic rats. The most important findings in this study are that FBP is a potent platelet aggregation inhibitor, in vitro and ex vivo. It presents protective effects on coagulation abnormalities, which can represent a treatment against DIC. The mechanisms for these effects remain under investigation.

An enzymatically inactivated hemoglobinase from Necator americanus induces neutralizing antibodies against multiple hookworm species and protects dogs against heterologous hookworm infection.

Hookworms digest hemoglobin from erythrocytes via a proteolytic cascade that begins with the aspartic protease, APR-1. Ac-APR-1 from the dog hookworm, Ancylostoma caninum, protects dogs against hookworm infection via antibodies that neutralize enzymatic activity and interrupt blood-feeding. Toward developing a human hookworm vaccine, we expressed both wild-type (Na-APR-1(wt)) and mutant (Na-APR-1(mut)-mutagenesis of the catalytic aspartic acids) forms of Na-APR-1 from the human hookworm, Necator americanus. Refolded Na-APR-1(wt) was catalytically active, and Na-APR-1(mut) was catalytically inactive but still bound substrates. Vaccination of canines with Na-APR-1(mut) and heterologous challenge with A. caninum resulted in significantly reduced parasite egg burdens (P=0.034) and weight loss (P=0.022). Vaccinated dogs also had less gut pathology, fewer adult worms, and reduced blood loss compared to controls but these did not reach statistical significance. Vaccination with Na-APR-1(mut) induced antibodies that bound the native enzyme in the parasite gut and neutralized enzymatic activity of Na-APR-1(wt) and APR-1 orthologues from three other hookworm species that infect humans. IgG1 against Na-APR-1(mut) was the most prominently detected antibody in sera from people resident in high-transmission areas for N. americanus, indicating that natural boosting may occur in exposed humans. Na-APR-1(mut) is now a lead antigen for the development of an antihematophagy vaccine for human hookworm disease.