Stages Of Sexual Development Research Articles

Indicators of dopamine system dysfunction and body mass index as prognostic markers for the of emotional status disturbances in pubertal age children

Objective. To create a prognostic algorithm of emotional disorders development in pubertal age children based on markers of dopamine system dysfunction and depending on body mass index and pubertal periods.Materials and methods. In the cross-sectional study Depression Self­RatingScale (DSRC) was used in 120 children aged from 11.6 to 17.9 years (the 2nd — 5th Tanner’s puberty stages). to establish depression risk (total DSRC score) and depressive symptoms (no depression; one depressive episode; presence of depression). Patients were stratified into groups BMI SDS: group 1 — children with normal weight, group 2 —those with simple obesity and group 3 — morbidly obese. All children were further divided into subgroups by gender and sexual development stage according to Tanner. Patients of 2 and 3 Tanner puberty stage were combined into early puberty (EP) subgroup, those of stage 4 and 5 — into late puberty (LP) subgroup Enzyme-linked immunosorbent assay (ELISA) was uses to identify blood dopamine (D) concentration in 88 patients. Two children with maximum and minimum D values were excluded from the study.D level quartiles were as follows: 1st—low concentration (< 10.9 pg/ml); 2nd — moderately decreased concentration (10.9—16.17 pg/ml); 3rd — moderately increased levels (16.17—19.3 pg/ml); 4th — high concentration (> 19.3 pg/ml). Statistical analysis was performed with SPSS Statistics 21 using Spearman’s nonparametric correlations (r), mathematical modeling method (logistic polynomial regression) (p < 0.05).Results and discussion. In the general group of children of LP age, the values of D and the indicators of emotional status formed a model with a statistically significant estimate (p = 0.016) to predict the probability of dopamine dysfunction. For example, a LP age patient with the presence of depression will have moderately decreased D concentration with 33.3 % probability, moderately elevated — with 16.7 %, and high D levels — with 50 % probability. In the subgroup of pubertal adolescent children with obesity, the values of the D ranges and the indicators of emotional status formed a model with a reliable statistical estimate (p = 0.05) to assess the probability of dopamine system dysfunction. For example, an obese adolescent with depression will have moderately reduced blood D level with 50 % probability; moderately elevated D — with 33.3 % and high D level in — 16.7 %. Based on these two models, we created the prognostic algorithm for diagnosing of dopamine system dysfunction in adolescents. It could be used as a tool for further scientific research in studying mechanisms of morbid obesity development in children.Conclusions. We developed prognostic math models with good statistical significance.It could be used to evaluate the effect of dopamine concentration ranges on the likelihood of depression presence, its single episode presence or its absence in adolescent children with obesity (p = 0.05) and in LP age patients (p = 0.016) regardless from body weight. Based on the developed mathematical models, we created the algorithm for diagnosing dopamine system dysfunction using BMI, puberty stage, and emotional status characteristics in adolescents.

A Multi-Stage Plasmodium vivax Malaria Vaccine Candidate Able to Induce Long-Lived Antibody Responses Against Blood Stage Parasites and Robust Transmission-Blocking Activity.

Malaria control and interventions including long-lasting insecticide-treated nets, indoor residual spraying, and intermittent preventative treatment in pregnancy have resulted in a significant reduction in the number of Plasmodium falciparum cases. Considerable efforts have been devoted to P. falciparum vaccines development with much less to P. vivax. Transmission-blocking vaccines, which can elicit antibodies targeting Plasmodium antigens expressed during sexual stage development and interrupt transmission, offer an alternative strategy to achieve malaria control. The post-fertilization antigen P25 mediates several functions essential to ookinete survival but is poorly immunogenic in humans. Previous clinical trials targeting this antigen have suggested that conjugation to a carrier protein could improve the immunogenicity of P25. Here we report the production, and characterization of a vaccine candidate composed of a chimeric P. vivax Merozoite Surface Protein 1 (cPvMSP1) genetically fused to P. vivax P25 (Pvs25) designed to enhance CD4+ T cell responses and its assessment in a murine model. We demonstrate that antibodies elicited by immunization with this chimeric protein recognize both the erythrocytic and sexual stages and are able to block the transmission of P. vivax field isolates in direct membrane-feeding assays. These findings provide support for the continued development of multi-stage transmission blocking vaccines targeting the life-cycle stage responsible for clinical disease and the sexual-stage development accountable for disease transmission simultaneously.

SUN-235 Obesity in Before and After Sexual Maturity All Led to Fertility Decline of Mice

Objective: As a mondial public health issue, obesity is deterministically regarded as an incentive to the testicular function decrease for the adults, whose effects on the adolescence remain unclear. By feeding the mice before and after their sexual maturation, this study exposes the high-fat diet effects on the testicular function in those two sexual development stages. Methods:The 4-week-old C57BL/6 male pups were divided into four groups for feeding on average:the mice on a continuous normal diet from 4 weeks to 16 weeks served as control group(NN),a high-fat diet group before sexual maturity(HN),that is from 4 weeks to 8 weeks,a high fat diet group after sexual maturity(NH) ,that is from 8 weeks to 16 weeks,and a continuous high fat diet group(HH) from 4 weeks to 16 weeks.At the age of 8 weeks and 16 weeks, the male mice were mated with the female mice at the age of 8 weeks, respectively. And the rate of the pregnancies of female mice, the number and the weight of offspring were observed to assess the fertility of the male mice.The parameters of functions on testis, such as sperms counting, the structure of acrosome, testosterone in serum and testicular structure, were all evaluated at 8 and 16 weeks,respectively. Results: At 16 weeks, mice in the continuous high fat diet group(HH)and high fat diet group after sexual maturity(NH) were compared with those in the control group(NN).Mice in the NH and HH group, the body weights had increased significantly, blood lipids had increased, the accumulation of testis lipid had increased, fat of epididymis has increased, apoptosis of testicular cells had increased, the number of sperms had decreased, acrosomal reactions of sperms had increased, testosterone of serum had not change significantly. Moreover,the fertility of male mice declined, showing a decrease in the pregnancies rate of female mice(NN group100% VS HH group 33.3%, NN group100% VS NH group 42%) and the average weight of offspring.(NN group 11.1±1.65 VS NH group7.00±1.08, NN group11.1±1.65 VS HH group 6.50±2.04.) Conclusions:In spite of starting feeding high-fat diet before adolescence of mice or starting feeding high-fat diet after adolescence of mice,as well as sustained high-fat diet mice group, the fertility and testicular function of male mice decreased compared with of the normal-diet group. Keywords:High-fat diets; Obesity; Testis

Re-evaluation of merogony of a Cystoisospora ohioensis-like coccidian and its distinction from gametogony in the intestine of a naturally infected dog.

Four species of Cystoisospora, C. canis, C. ohioensis, C. neorivolta and C. burrowsi are described from feces of dogs. Of these, the oocysts of C. canis are the largest and easily distinguished from the remaining three species. Oocysts of C. ohioensis, C. neorivolta and C. burrowsi are difficult to distinguish because of overlap in their sizes. However, based on endogenous developmental stages, C. ohioensis is distinct from C. neorivolta and C. burrowsi because its endogenous stages are confined to surface epithelium of intestine whereas endogenous stages of C. neorivolta and C. burrowsi are predominantly in the lamina propria. There are uncertainties regarding the endogenous stages of C. neorivolta and C. burrowsi and there is no way now to determine whether C. burrowsi and C. neorivolta are different parasites; therefore, these are referred as C. ohioensis-like organisms. Additionally, mode of division of asexual stages of coccidia of dogs is largely unknown and ultrastructural studies are lacking. In the present study, development of asexual and sexual stages of a C. ohioensis-like organism in a naturally infected dog is described by light microscopy and by transmission electron microscopy. Merozoites divided by endodyogeny/merogony. Meronts were crescent/merozoite-shaped and contained a maximum of eight nuclei. A distinctive feature of merozoites was the presence of many PAS-positive amylopectin granules that were absent or rare in immature microgamonts making it possible to distinguish them.

MilR4 and milR16 Mediated Fruiting Body Development in the Medicinal Fungus Cordyceps militaris.

Cordyceps militaris readily performs sexual reproduction, thus providing a remarkably rich model for understanding the processes involved in sexual development. It could regulate expression of human genes by diet-derived miRNA-like RNAs (milRNAs). However, the study of miRNAs in C. militaris has been limited. In the present study, genes encoding Dicers, Argonautes, and RNA-dependent RNA polymerases were identified. Illumina deep sequencing was performed to characterize the milRNAs in C. militaris at asexual and sexual development stages. Total 38 milRNAs were identified and five milRNAs were validated by northern blot and qRT-PCR, out of which, 19 were specific for sexual development. Importantly, the fungi could not form fruiting bodies after disruption of milR4, while the perithecium was formed in advance after over-expression of milR4. Abnormal pale yellow fruiting body primordium, covered with abnormal primordium, was formed in the strain with miR16 disruption. Although no milR4 or milR16 target genes were identified, differential expression of many different genes involved in mycelium growth and sexual development (mating process, mating signaling, and fruiting body development) among these mutants were found. Overall, milRNAs play vital roles in sexual development in C. militaris.

Transgenic Babesia bovis lacking 6-Cys sexual-stage genes as the foundation for non-transmissible live vaccines against bovine babesiosis

Babesia bovis, a tick-borne apicomplexan parasite responsible for bovine babesiosis has a complex life cycle including sexual development in its Rhipicephalus microplus vector. Understanding the molecular mechanisms involved in sexual development is essential for developing future-generation transmission blocking vaccines (TBVs) and/or non-transmissible attenuated live vaccines. The widely conserved members of the 6-Cys gene family likely play roles in the development of sexual stages of B. bovis, and are candidates for developing novel TBV. The recently defined sexual markers 6-CysA and 6-CysB of B. bovis are strain-conserved and exclusively surface-expressed in tick-stage parasites. However, the high level of sequence identity among the 6-Cys A and 6-Cys B proteins (52% identity), together with similar 6-Cys domain distribution and sub-cellular localization, are suggestive of redundant function. We hypothesized that disruption of both 6-CysA and 6-CysB in B. bovis would result in unaltered ability of the parasite to invade and grow in red blood cells (RBCs), with concomitant loss of the transmission phenotype. Taking advantage of their contiguous genome localization, we generated a double gene-knockout system to disrupt a 3287 bp region encompassing both 6-CysA and 6-CysB genes using a single transfection plasmid. The resulting red-fluorescent ΔAΔB 6-Cys B. bovis transgenic parasite line was able to grow continuously in bovine RBCs in vitro at a similar rate to wild-type parasites, demonstrating that the 6-CysA and 6-CysB genes are not required for the development of blood-stage parasites. This novel gene manipulation approach will allow future experiments aimed at determining the tick-transmission phenotype of parasites lacking tick-stage genes. Parasites deficient in genes required for sexual reproduction could be the foundation for genetically-defined, non-transmissible live vaccines against bovine babesiosis. Developing a non-tick transmissible live vaccine based on attenuated parasites unable to express critical 6-Cys genes and including a molecular vaccine marker could help reduce the burden of bovine babesiosis globally.

Proteomic analysis of Tianzhu White Yak (Bos grunniens) testis at different sexual developmental stages.

To explore the protein expression profiles of white yak (Bos grunniens) testis at different sexual developmental stages. The protein profiles of yak testis were determined using two-dimensional electrophoresis and the expression levels of 298 protein spots were analyzed. Mass spectrometry was performed to identify those significantly differential expressed proteins; Western blotting was used to confirm the results. During the developmental stages, 29 protein spots showed more than twofold differences (p<0.05) at ≥1 time point and were successfully identified. Two proteins were upregulated with age (category 1), five proteins (17.2%) were downregulated with age (category 2), four proteins were upregulated before 4years of age and downregulated thereafter (category 3), fifteen proteins were upregulated before 2years of age and downregulated thereafter (category 4), and three proteins fluctuated with age (category 5). The expression patterns of regucalcin and heat shock 60kDa protein in category 2 were confirmed. The 29 differentially expressed proteins from yak testes (some had more than one function) were categorized into binding (n=15), catalytic activity (n=13), molecular function regulator (n=4), antioxidant (n=4), molecular transducer (n=2), transporter (n=1), and structural molecule (n=1). The identification and analysis of these testis proteins may assist in understanding the developmental biology of reproduction system in male yak.

Anti-plasmodial Activity of a Non-protein Amino Acid Taurine

Human malaria is caused by a few selected species of the genus Plasmodium. Among these, Plasmodium falciparum causes almost 90% of malaria-related mortality. Novel anti-malarial compounds are hence required to fight the anti-malarial drug-resistant P. falciparum parasites. The objective of this study is to analyze the effectiveness of Taurine (2-aminoethane sulfonic acid), a non-protein amino acid, in preventing the growth and development of both asexual and sexual stages of in vitro cultured P. falciparum parasites. We found that 200 mM concentration of Taurine almost completely (>80%) inhibited the propagation of asexual stages of P. falciparum. In contrast, it did not have any inhibitory activity against the maturation of sexual or gametocyte stages. However, the gametocytogenesis or the conversion of asexual to stage I gametocyte was blocked partially by this compound. The results suggest that derivatives of Taurine /2-aminoethane sulfonic acid could be considered to further improve the effectiveness of Taurine as an antimalarial compound against both the asexual and early sexual stages of P. falciparum.

Age, growth and reproductive biology of the tub gurnard (Chelidonichthys lucerna) in North-East Portugal

Event Abstract Back to Event Age, growth and reproductive biology of the tub gurnard (Chelidonichthys lucerna) in North-East Portugal Juliana Rodrigues1, 2*, Diana Feijó3, Alberto Rocha3, Karim Erzini1, 4 and Alberto T. Correia2, 5* 1 University of Algarve, Portugal 2 Interdisciplinary Center for Marine and Environmental Research, Abel Salazar Institute of Biomedical Sciences, University of Porto, Portugal 3 Portuguese Institute of Ocean and Atmosphere (IPMA), Portugal 4 Centre of Marine Sciences, University of Algarve, Portugal 5 Fernando Pessoa University, Portugal The tub gurnard, Chelidonichthys lucerna, is an important commercial marine demersal fish species that belongs to Triglidae which can be commonly found in the north-eastern Atlantic, in the Mediterranean Sea and in the Black Sea. C. lucerna is mainly caught through bottom and beam trawls, gillnets, trammel nets and longlines. Within the gurnards, C. lucerna is the most important and the one with the greatest economic value in the Portuguese coast. The aim of the study is to obtain, for the fisrt time, data on the age, growth, sex ratio and sexual developmental stages of C. lucerna captured on the northern coast of Portugal (Matosinhos) bimonthly between June 2018 and May 2019. Moreover, we intent to test whether these basic biological traits, at present unknown for this species in the southern area of the NE Atlantic, differ from other geographic locations. These new findings are important to a sustainable and rational fisheries management. Acknowledgements This work was supported by FCT - Foundation for Science and Technology - (SFRH/BD/111133/2015 to CM by NORTE-01-0145-FEDER-000035 (MARINFO) and the Strategic Funding UID/Multi/04423/2019 through national funds provided by FCT and European Regional Development Fund (ERDF), in the framework of the programme PT2020. Keywords: Triglidae, Fisheries, life history traits, age, Reproductive biology Conference: XX Iberian Symposium on Marine Biology Studies (SIEBM XX) , Braga, Portugal, 9 Sep - 12 Sep, 2019. Presentation Type: Poster Presentation Topic: Fisheries, Aquaculture and Biotechnology Citation: Rodrigues J, Feijó D, Rocha A, Erzini K and Correia AT (2019). Age, growth and reproductive biology of the tub gurnard (Chelidonichthys lucerna) in North-East Portugal. Front. Mar. Sci. Conference Abstract: XX Iberian Symposium on Marine Biology Studies (SIEBM XX) . doi: 10.3389/conf.fmars.2019.08.00158 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 17 May 2019; Published Online: 27 Sep 2019. * Correspondence: Mx. Juliana Rodrigues, University of Algarve, Faro, Portugal, julianacrfrodrigues@gmail.com Prof. Alberto T Correia, Interdisciplinary Center for Marine and Environmental Research, Abel Salazar Institute of Biomedical Sciences, University of Porto, Matosinhos, 4450-208, Portugal, atcorreia.ciimar@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Juliana Rodrigues Diana Feijó Alberto Rocha Karim Erzini Alberto T Correia Google Juliana Rodrigues Diana Feijó Alberto Rocha Karim Erzini Alberto T Correia Google Scholar Juliana Rodrigues Diana Feijó Alberto Rocha Karim Erzini Alberto T Correia PubMed Juliana Rodrigues Diana Feijó Alberto Rocha Karim Erzini Alberto T Correia Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Pengetahuan Orangtua Mengenai Pendidikan Seksual Bagi Anak dengan Gangguan Spektrum

Abstract - Parents play an important role in sexual education for their children. The role also owned by parents who have children with autism spectrum disorder (ASD). However, information about sexual education for children with ASD is limited. This condition caused parents feel confused about when and how to provide sexual education to their children who have ASD. This study aims to determine the level of parental knowledge about sexual education in children with ASD. The measuring instrument used in this study was a test of knowledge about sexual education in children with ASD. Respondents in this study involved 80 parents who had children with ASD in Surabaya. The results showed that 28% of respondents had a moderate level of knowledge regarding sexual education in children with ASD. Nearly half or 48% of respondents have a good level of knowledge, and 15% of respondents have a lack of knowledge. Most parents have insufficient knowledge in terms of understanding sexuality, the stage of sexual development, and the topic of sexual education that is appropriate for the age of the children with ASD. Keywords: sexual education, parental knowledge, autism spectrum disorder Abstrak – Orangtua memegang peranan penting dalam pendidikan seksual bagi anaknya. Termasuk orangtua yang memiliki anak dengan gangguan spektrum autis (GSA). Akan tetapi, informasi tentang pendidikan seksual bagi anak dengan GSA ini terbatas. Kondisi tersebut mengakibatkan orangtua merasa bingung tentang kapan dan bagaimana memberikan pendidikan seksual pada anaknya yang memiliki GSA. Penelitian ini bertujuan untuk mengetahui tingkat pengetahuan orangtua tentang pendidikan seksual pada anak GSA. Alat ukur yang digunakan dalam penelitian ini adalah tes pengetahuan tentang pendidikan seksual pada anak GSA. Responden dalam penelitian ini adalah 80 orangtua yang memiliki anak GSA di Surabaya. Hasil penelitian menunjukkan bahwa 28% responden memiliki tingkat pengetahuan yang sedang mengenai pendidikan seksual pada anak dengan GSA. Hampir separuh atau 48% responden memiliki tingkat pengetahuan yang baik, dan 15% responden memiliki tingkat pengetahuan yang kurang. Sebagian besar orangtua memiliki pengetahuan yang kurang dalam aspek pengertian seksualitas, tahap perkembangan seksual, dan topik pendidikan seksual yang sesuai dengan usia anak GSA. Kata kunci: pendidikan seksual, pengetahuan orangtua, gangguan spektrum autisÂ

Characteristics of physical development of school-age children affected by the armed conflict in eastern Ukraine

Objective — to determine characteristics of disorders in physical development (PD) of school-age children affected by the armed conflict in eastern Ukraine.Materials and methods. We assessed the nature of PD of children and adolescents aged 6—18 years (179 girls and 128 boys) and affected by the armed conflict in eastern Ukraine, who had applied to the State institution «Institute for Children and Adolescents Health Care of the National Academy of Medical Sciences of Ukraine» for medical help in 2015—2018. Taking into account age and stage of sexual development as of the beginning of Anti-Terrorist Operation (ATO), the following groups were formed: childhood (6—8 years); prepuberty (9—10 years); early puberty (11—13 years); puberty itself (14—16 years); late puberty (17—18 years). Software packages «SPSS Statistics 17.0» and «Excel» were used for mathematical processing of received results.Results and discussion. We established causality between nature of PD, sex and age as of the beginning of the armed conflict (April 2014). In remote catamnesis we identified disharmonious PD the most frequently in children, who in 2014 had been less than 7 years old (66.7 %), by low height in boys and by excess weight in girls. In contrast, in boys, who in 2014 had been 9—10 years old, we identified excess body weight twice as frequently as in girls of the same age (42.9 % and 27.6 %, respectively). This age group is the one in dynamics of monitoring of which we diagnosed complicated progression of obesity. It was determined that excess weight had been the most frequent form of PD disorder in adolescents of both sexes. Body mass deficiency was identified much less frequently, mostly in boys, who in 2014 had been 11–13 years old (14.6 %).Conclusions. The risk group for PD disorders is schoolchildren affected by the armed conflict at younger age and at the onset of puberty, in whose pathology structure excess body mass had been prevalent.

Heterochronic reproductive developmental processes between diploid and tetraploid cytotypes of Paspalum rufum.

Apomixis is an asexual reproductive mode via seeds that generate maternal clonal progenies. Although apomixis in grasses is mainly expressed at the polyploid level, some natural diploid genotypes of Paspalum rufum produce aposporous embryo sacs in relatively high proportions and are even able to complete apomixis under specific conditions. However, despite the potential for apomixis, sexuality prevails in diploids, and apomixis expression is repressed for an as yet undetermind reason. Apomixis is thought to derive from a deregulation of one or a few components of the sexual pathway that could be triggered by polyploidy and/or hybridization. The objectives of this work were to characterize and compare the reproductive development and the timing of apospory initial (AI) emergence between diploid genotypes with potential for apomixis and facultative apomictic tetraploid cytotypes of P. rufum. Reproductive characterization was performed by cytoembryological observations of cleared ovaries and anthers during all reproductive development steps and by quantitative evaluation of the ovule growth parameters. Cytoembryological observations showed that in diploids, both female and male reproductive development is equally synchronized, but in tetraploids, megasporogenesis and early megagametogenesis are delayed with respect to microsporogenesis and early microgametogenesis. This delay was also seen when ovary growth was taken as a reference parameter. The analysis of the onset of AIs revealed that they emerge during different developmental periods depending on the ploidy level. In diploids, the AIs appeared along with the tetrad (or triad) of female meiocytes, but in tetraploids they appeared earlier, at the time of the megaspore mother cell. In both cytotypes, AIs can be seen even during megagametogenesis. Overall observations reveal that female sexual reproductive development is delayed in tetraploids as compared with diploid genotypes, mainly at meiosis. In tetraploids, AIs appear at earlier sexual developmental stages than in diploids, and they accumulate up to the end of megasporogenesis. The longer extension of megasporogenesis in tetraploids could favour AI emergence and also apomixis success.

An unusual sexual stage in the alkalophilic ascomycete Sodiomyces alkalinus

Exploring life cycles of fungi is insightful for understanding their basic biology and can highlight their ecology. Here, we dissected the sexual and asexual life cycles of the obligate alkalophilic ascomycete Sodiomyces alkalinus that thrives at extremely high pH of soda lakes. S.alkalinus develops acremonium-type asexual sporulation, commonly found in ascomycetous fungi. However, the sexual stage was unusual, featuring very early lysis of asci which release young ascospores inside a fruit body long before its maturation. In a young fruit body, a slimy matrix which originates from the combined epiplasm of asci and united cytoplasm of the pseudoparenchymal cells, surrounds pooled maturing ascospores. Upon maturity, the ascospores are forcibly released through a crack in the fruit body, presumably due to an increased turgor pressure. These features of the sexual stage development resemble the ones found in unrelated marine fungi, indicating convergent evolution of the trait. We hypothesise these developmental features of S. alkalinus to be adaptive in the conditions of periodically inundated rims of soda lakes where the fungus thrives.

The use of recombinant FSH in combination therapy in a young male with idiopathic hypogonadotropic hypogonadism

Currently, testosterone drugs are used to treat hypogonadotropic hypogonadism, which allow men to get a good height and reach a stage of sexual development corresponding to their age. In this case, the testicular volume remains of pre-pubertal size, and the spermatogenic epithelium fails to reach its mature stage under such conditions. The study was aimed at initiating puberty in a 17-year-old male with hypogonadotropic hypogonadism using gonadotropic hormone drugs. The patient received foliotropinalpha injections in combination with chorionic gonadotropin for 9 months. The treatment efficacy was evaluated after 3, 6, 9 months of therapy by measuring the testicle volume (using Prader orchidometer and ultrasound) and the sex hormones and inhibin B serum levels. During the treatment period, the testicle volume increased from 1.5 to 8 ml based on clinical measurements, and from 1.38 and 1.14 to 5.8 and 5.87 ml (right and left, respectively) based on ultrasound imaging. The level of testosterone reached normal values, inhibin B also increased. The use of recombinant FSH for nine months in the combination therapy of idiopathic hypogonadotropic hypogonadism in a 17-year-old male contributed to the initiation of a true puberty.

Identification of a putative methyltransferase gene of Babesia bigemina as a novel molecular biomarker uniquely expressed in parasite tick stages

BackgroundBovine babesiosis is caused by apicomplexan pathogens of the genus Babesia such as B. bigemina and B. bovis. These tick-borne pathogens have a complex life-cycle involving asexual multiplication in vertebrate hosts and sexual reproduction in invertebrate vectors. In the tick midgut, extracellular Babesia parasites transform into gametes that fuse to form zygotes. Understanding the mechanisms that underlie formation of extracellular Babesia tick stages is an important step towards developing control strategies for preventing tick infection and subsequent parasite transmission.ResultsWe induced B. bigemina sexual stages in vitro by exposing parasites to Tris 2-carboxyethyl phosphine (TCEP). Subsequently, we identified a novel putative methyltransferase gene (BBBOND_0204030) that is expressed uniquely in all B. bigemina tick stages but not in blood stages. In vitro TCEP-exposed B. bigemina presented diverse morphology including parasites with long projections, round forms and clusters of round forms indicative of sexual stage induction. We confirmed the development of sexual stages by detecting upregulation of previously defined B. bigemina sexual stage marker genes, ccp2 and 3, and their respective protein expression in TCEP-induced B. bigemina cultures. Next, transcription analysis of in vitro TCEP-induced B. bigemina culture based on an in silico derived list of homologs of Plasmodium falciparum gamete-specific genes demonstrated differential expression of the gene BBBOND_0204030 in induced cells. Further examination of ex vivo infected ticks demonstrated that BBBOND_0204030 is transcribed by multiple stages of B. bigemina during parasite development in tick midgut, ovary and hemolymph. Interestingly, ex vivo results confirmed our in vitro observation that blood stages of B. bigemina do not express BBBOND_0204030 and validated the in vitro system of inducing sexual stages.ConclusionsHerein we describe the identification of a B. bigemina gene transcribed exclusively by parasites infecting ticks using a novel method of inducing B. bigemina sexual stages in vitro. We propose that this gene can be used as a marker for parasite development within the tick vector. Together, these tools will facilitate our understanding of parasite-tick interactions, the identification of novel vaccine targets and, consequently, the development of additional strategies to control bovine babesiosis.

Chimeric Plasmodium falciparum parasites expressing Plasmodium vivax circumsporozoite protein fail to produce salivary gland sporozoites

BackgroundRodent malaria parasites where the gene encoding circumsporozoite protein (CSP) has been replaced with csp genes from the human malaria parasites, Plasmodium falciparum or Plasmodium vivax, are used as pre-clinical tools to evaluate CSP vaccines in vivo. These chimeric rodent parasites produce sporozoites in Anopheles stephensi mosquitoes that are capable of infecting rodent and human hepatocytes. The availability of chimeric P. falciparum parasites where the pfcsp gene has been replaced by the pvcsp would open up possibilities to test P. vivax CSP vaccines in small scale clinical trials using controlled human malaria infection studies.MethodsUsing CRISPR/Cas9 gene editing two chimeric P. falciparum parasites, were generated, where the pfcsp gene has been replaced by either one of the two major pvcsp alleles, VK210 or VK247. In addition, a P. falciparum parasite line that lacks CSP expression was also generated. These parasite lines have been analysed for sporozoite production in An. stephensi mosquitoes.ResultsThe two chimeric Pf-PvCSP lines exhibit normal asexual and sexual blood stage development in vitro and produce sporozoite-containing oocysts in An. stephensi mosquitoes. Expression of the corresponding PvCSP was confirmed in oocyst-derived Pf-PvCSP sporozoites. However, most oocysts degenerate before sporozoite formation and sporozoites were not found in either the mosquito haemocoel or salivary glands. Unlike the chimeric Pf-PvCSP parasites, oocysts of P. falciparum parasites lacking CSP expression do not produce sporozoites.ConclusionsChimeric P. falciparum parasites expressing P. vivax circumsporozoite protein fail to produce salivary gland sporozoites. Combined, these studies show that while PvCSP can partially complement the function of PfCSP, species-specific features of CSP govern full sporozoite maturation and development in the two human malaria parasites.

Micropenis

Micropenis refers to a normally structured penis, which is abnormally small, with a stretched length of less than 2.5 standard deviations (SD) below the mean of age or stage of sexual development. Patients with true micropenis usually have a 46,XY karyotype, associated to normally located male gonads, without any sign of other penile malformations. Penile length is measured from the point where the penis meets the pubic bone to the distal tip of the glans on maximal stretch, compressing any fat over suprapubic area. Differential diagnosis include buried penis (inability to adequately expose the penis, due to a severe phymosis associated to a decreased length of penile skin, which retain the shaft inside the preputial cavity), and obesity with excess fat pad over supra pubic area. The causes of congenital or true micropenis can be divided into three major groups: hypogonadotropic hypogonadism (pituitary/hypothalamic failure), hypergonadotropic hypogonadism (primary testicular failure) and idiopathic (associated with a functional hypothalamus–pituitary–testicular axis). Clinical treatment is based on testosterone therapy. Surgical treatment options are used after failed penile enlargement with testosterone therapy and include penile elongation, suprapubic fat pad aspiration and eventually neophaloplasty.

Characterization of female sexual development‐1 (fsd‐1) Transcript Structure, Expression, and Localization in the Fungus Neurospora crassa

Neurospora crassa is a filamentous ascomycete fungus that can reproduce sexually and asexually. The signaling pathways that control sexual development in N. crassa are not well characterized. FSD‐1 is an N. crassa transcription factor that controls both early and late stages of sexual development. Although we know that fsd‐1 regulates female sexual development (including fertility and spore maturation), little is known about (1) its transcriptional regulation, (2) RNA isoforms, (3) protein expression, and (4) localization. Intron and exon boundaries of fsd‐1 were mapped using reverse transcription (RT)‐PCR and sequencing and we determined that fsd‐1 can be expressed as three different transcripts, which differ in length and intron/exon structure. Quantitative RT‐PCR during a time course of sexual development showed that expression of fsd‐1 transcripts is not detectable until N. crassa initiates sexual development. In addition, one transcript variant is predominantly expressed during sexual development, and expression of this transcript peaks approximately five days after crossing (mating). It is unknown whether all three fsd‐1 transcripts are translated into protein, and thus we will monitor FSD‐1 protein expression using immunoprecipitation and western blotting. The three fsd‐1 transcript variants encode for predicted proteins of slightly different sizes, which can be visualized via western blot. To investigate FSD‐1 localization, we constructed a GFP‐tagged version of FSD‐1 using overlap PCR and targeted this construct to the native locus. FSD‐1‐GFP localization was monitored via fluorescence microscopy throughout a time course of development. We found that FSD‐1‐GFP localized to the nuclei of female reproductive tissues, such as paraphyses, but not to tissues where meiosis occurs. In addition, we confirmed that the FSD‐1‐GFP strain complements the sexual development‐related phenotypes that occur in an fsd‐1 knockout strain, such as sterility and lack of ascospore melanization. Finally, we constructed strains where fsd‐1 has an overexpression promoter, and we will determine whether overexpression of fsd‐1 has a dominant negative effect on wild‐type strains during mating. Little is known about the regulatory pathways that function in non‐meiotic filamentous fungal female sexual tissues, and our work on fsd‐1 provides an important first step in characterizing this aspect of fungal development.Support or Funding InformationGeneseo FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

PATHOLOGICAL AND PARASITOLOGICAL STUDIES ON SOME EIMERIA SPECIES IN RABBITS USING LIGHT AND ELECTRON MICROSCOPE

The prevalence of Eimeria in 50 examined rabbits was 84% (42/50). The prevalence was 92.6% in male rabbits while it was73.9% in female rabbits. According to age, the prevalence was determined 88% in rabbits less than 4 months of age while it was 80% in rabbits 4 months of age or more. No significant difference was recorded between rabbits in correlation to age and sex. Ten species of Eimeria infecting rabbits were isolated in Assiut by parasitological examination. The prevalence of Eimeria species were E. perforans (66.7%) followed by E. exigua (26.2%), E. media (26.2%), E. magna (21.4%), E. intestinalis (19%), E. coecicola (19%), E. irresidua (19%), E. piriformis (14.3%), E. flavescens (7.1%) and E. stiedae (7.1%). Single infection of Eimeria spp. was found in 23.8% of the infected rabbits, where as mixed infection involved two, three or four Eimeria spp. was observed in 76.2% of the infected rabbits. Clinical signs were depression, anorexia, diarrhea while postmortem examination revealed hepatomegaly with presence of separate yellowish-white nodules of varying sizes spread over the surface with distended gall bladder. The intestinal lesions revealed varying degree of congestion, thickening of intestinal wall. Histopathological examination of the liver revealed dilated bile ducts and formation of papilliform projections of epithelium containing different developmental stages of Eimeria, associated with degeneration and pathological changes in hepatic parenchyma. The intestinal coccidiosis revealed hyperplasia of the epithelial cells and presence of Eimeria oocytes and gametocytes within the epithelial cells of the villi associated with lymphocytic infiltration in the lamina propria of the villi. The transmission electron microscope showed asexual and sexual developmental stages of rabbit Eimeria including developing schizont and macrogametocyte.

Rhipicephalus appendiculatus ticks transmit Theileria parva from persistently infected cattle in the absence of detectable parasitemia: implications for East Coast fever epidemiology

BackgroundEast Coast fever (ECF) is a devastating disease of cattle and a significant constraint to improvement of livestock production in sub-Saharan Africa. The protozoan parasite causing ECF, Theileria parva, undergoes obligate sexual stage development in its tick vector Rhipicephalus appendiculatus. Tick-borne acquisition and transmission occurs transstadially; larval and nymphal ticks acquire infection while feeding and transmit to cattle when they feed after molting to the next stage. Much of the current knowledge relating to tick-borne acquisition and transmission of T. parva has been derived from studies performed during acute infections where parasitemia is high. In contrast, tick-borne transmission during the low-level persistent infections characteristic of endemic transmission cycles is rarely studied.MethodsCattle were infected with one of two stocks of T. parva (Muguga or Marikebuni). Four months post-infection when parasites were no longer detectable in peripheral blood by PCR, 500 R. appendiculatus nymphs were fed to repletion on each of the cattle. After they molted to the adult stage, 20 or 200 ticks, respectively, were fed on two naïve cattle for each of the parasite stocks. After adult ticks fed to repletion, cattle were tested for T. parva infection by nested PCR and dot blot hybridization.ResultsOnce they had molted to adults the ticks that had fed as nymphs on Muguga and Marikebuni infected cattle successfully transmitted Theileria parva to all naïve cattle, even though T. parva infection was not detectable by nested PCR on salivary gland genomic DNA of a sample of individual ticks. However, a salivary gland homogenate from a single Marikebuni infected tick was able to infect primary bovine lymphocytes. Infection was detected by nested p104 PCR in 3 of 4 calves and detected in all 4 calves by T. parva 18S nested PCR/dot blot hybridization.ConclusionWe show that R. appendiculatus ticks are able to acquire T. parva parasites from infected cattle even in the absence of detectable parasitemia. Although infection was undetectable in a sample of individual ticks, cumulatively as few as 20 ticks were able to transmit T. parva to naïve cattle. These results have important implications for our understanding of T. parva transmission by R. appendiculatus in ECF endemic regions.