Stages Of Sexual Development Research Articles

Heterochromatin dynamics during the initial stages of sexual development in Plasmodium falciparum

Asexual replication of Plasmodium falciparum in the human blood results in exponential parasite growth and causes all clinical symptoms of malaria. However, at each round of the replicative cycle, some parasites convert into sexual precursors called gametocytes, which develop through different stages until they become infective to mosquito vectors. The genome-wide distribution of heterochromatin, a type of chromatin generally refractory to gene expression, is identical at all asexual blood stages, but is altered in stage II/III and more mature gametocytes. However, it is not known if these changes occur concomitantly with sexual conversion or at a later time during gametocyte development. Using a transgenic line in which massive sexual conversion can be conditionally induced, we show that the genome-wide distribution of heterochromatin at the initial stages of sexual development (i.e., sexual rings and stage I gametocytes) is almost identical to asexual blood stages, and major changes do not occur until stage II/III. However, we found that at loci with heterochromatin alterations, transcriptional changes associated with sexual development typically precede, rather than follow, changes in heterochromatin occupancy.

Peculiarities of the menstrual cycle in girls living in the Murmansk region

The aim of the study. To assess the characteristics of the menstrual cycle and ovulation disorders in girls aged 15–17 years.Materials and methods. The study was carried out in 2022–2023 in the Murmansk region in Apatity. A total of 225 girls with average age of 16.5 ± 0.03 years took part in the study. The characteristics of the menstrual cycle and ovulation disorders were studied according to the guidelines of the International Federation of Gynecology and Obstetrics.Results of the study. In the study group, the onset of menarche (13.1 years) is observed at a later age compared to the girls living in other regions of Russia and neighboring countries (12.6–12.9 years). Every seventh girl has an irregular menstrual cycle, every fifth girl has a cycle duration of more than 38 days. Heavy menstruation occurs in every third girl, intermenstrual uterine bleeding – in every thirteenth girl. Half of the girls have painful periods (dysmenorrhea). Among ovulation disorders, the most common is abnormal uterine bleeding – 34.7 %. Also, intermenstrual uterine bleeding (8.0 %), primary amenorrhea (2.7 %) and polycystic ovary syndrome (1.3 %) are registered. Among gynecological pathology, in addition to ovulation disorders, we registered vulvovaginal candidiasis and paraurethral cyst.Conclusion. The analysis of the obtained results showed that only 20.4 % of girls do not have gynecological pathology, which to a certain extent indicates a low level of reproductive health of girls. Asaresult, it is necessary to diagnose and treat diseases which can cause the development of ovulation disorders more carefully and at earlier stages of sexual development. The work was carried out as part of the research topic No. FMEZ-2022-0012.

Investigating Development and Defense Systems in Early Reproductive Stages of Male and Female Gonads in Black Scorpionfish Scorpaena porcus (Linnaeus, 1758).

One of the most crucial biological indicators in tracking long-term variations in the reproductive cycle is sexual development. Scorpaena porcus (Linnaeus, 1758), commonly known as the black scorpionfish, is a small teleost from the family Scorpaenidae. Much is known about its ecology, but data on its reproductive and defense systems are still lacking. Antimicrobial peptides (AMPs), such as piscidins, are integral components of the innate immune system in fish. These peptides exhibit a wide range of activity against bacteria, fungi, viruses, and protozoa and act as the first line of host defense. This study aims to investigate the primary sexual development stages in male and female gonads of black scorpionfish, providing additional knowledge on the reproductive biology of this teleost while evaluating concomitant changes in the expression of a Piscidin-1 antimicrobial peptide. The results show a histological, morpho-structural change from the immature stage to the developing virgin stage. Immunohistochemical analyses show that germinal and somatic cells are strongly reactive to Piscidin-1 in both gonads at an early ontogeny stage. These data suggest that Piscidin-1 may play a key role in the local defense system of scorpionfish gonads at this delicate stage, which is critical for the continuation and maintenance of the species. The present findings are potentially useful for a better understanding of the reproductive cycle of this fish, improving our knowledge of the interaction between the immune system and reproduction.

RebL1 is required for macronuclear structure stability and gametogenesis in Tetrahymena thermophila

Histone modification and nucleosome assembly play important roles in chromatin-related processes. Histone chaperones form different complexes and coordinate histone transportation and assembly. Various histone chaperone complexes have been identified in different organisms. The ciliate protozoa (ciliates) have various chromatin structures and different nuclear morphology. However, histone chaperone components and functions of different subunits remain unclear in ciliates. Tetrahymema thermophila contains a transcriptionally active macronucleus (MAC) and a transcriptionally inactive micronucleus (MIC) which exhibit multiple replication and various chromatin remodeling progresses during vegetative growth and sexual developmental stages. Here, we found histone chaperone RebL1 not only localized evenly in the transcriptionally active MAC but also dynamically changed in the MIC during vegetative growth and sexual developmental stages. REBL1 knockdown inhibited cellular proliferation. The macronuclear morphology became bigger in growing mutants. The abnormal macronuclear structure also occurred in the starvation stage. Furthermore, micronuclear meiosis was disturbed during sexual development, leading to a failure to generate new gametic nuclei. RebL1 potentially interacted with various factors involved in histone-modifying complexes and chromatin remodeling complexes in different developmental stages. REBL1 knockdown affected expression levels of the genes involved in chromatin organization and transcription. Taken together, RebL1 plays a vital role in maintaining macronuclear structure stability and gametogenesis in T. thermophila.

Plasmodium falciparum contains functional SCF and CRL4 ubiquitin E3 ligases, and CRL4 is critical for cell division and membrane integrity.

Protein ubiquitination is essential for cellular homeostasis and regulation of several processes, including cell division and genome integrity. Ubiquitin E3 ligases determine substrate specificity for ubiquitination, and Cullin-RING E3 ubiquitin ligases (CRLs) make the largest group among the ubiquitin E3 ligases. Although conserved and most studied in model eukaryotes, CRLs remain underappreciated in Plasmodium and related parasites. To investigate the CRLs of human malaria parasite Plasmodium falciparum, we generated parasites expressing tagged P. falciparum cullin-1 (PfCullin-1), cullin-2 (PfCullin-2), Rbx1 (PfRbx1) and Skp1 (PfSkp1). PfCullin-1 and PfCullin-2 were predominantly expressed in erythrocytic trophozoite and schizont stages, with nucleocytoplasmic localization and chromatin association, suggesting their roles in different cellular compartments and DNA-associated processes. Immunoprecipitation, in vitro protein-protein interaction, and ubiquitination assay confirmed the presence of a functional Skp1-Cullin-1-Fbox (PfSCF) complex, comprising of PfCullin-1, PfRbx1, PfSkp1, PfFBXO1, and calcyclin binding protein. Immunoprecipitation, sequence analysis, and ubiquitination assay indicated that PfCullin-2 forms a functional human CRL4-like complex (PfCRL4), consisting of PfRbx1, cleavage and polyadenylation specificity factor subunit_A and WD40 repeat proteins. PfCullin-2 knock-down at the protein level, which would hinder PfCRL4 assembly, significantly decreased asexual and sexual erythrocytic stage development. The protein levels of several pathways, including protein translation and folding, lipid biosynthesis and transport, DNA replication, and protein degradation were significantly altered upon PfCullin-2 depletion, which likely reflects association of PfCRL4 with multiple pathways. PfCullin-2-depleted schizonts had poorly delimited merozoites and internal membraned structures, suggesting a role of PfCRL4 in maintaining membrane integrity. PfCullin-2-depleted parasites had a significantly lower number of nuclei/parasite than the normal parasites, indicating a crucial role of PfCRL4 in cell division. We demonstrate the presence of functional CRLs in P. falciparum, with crucial roles for PfCRL4 in cell division and maintaining membrane integrity.

RNA-seq analysis of parasitism by Intoshia linei (Orthonectida) reveals protein effectors of defence, communication, feeding and growth.

Orthonectida is a group of multicellular endoparasites of a wide range of marine invertebrates. Their parasitic stage is a multinuclear shapeless plasmodium infiltrating host tissues. The development of the following worm-like sexual generation takes place within the cytoplasm of the plasmodium. The existence of the plasmodial stage and the development of a sexual stage within the plasmodium are unique features to Bilateria. However, the molecular mechanisms that maintain this peculiar organism, and hence enable parasitism in orthonectids, are unknown. Here, we present the first-ever RNA-seq analysis of the plasmodium, aimed at the identification and characterization of the plasmodium-specific protein-coding genes and corresponding hypothetical proteins that distinguish the parasitic plasmodium stage from the sexual stage of the orthonectid Intoshia linei Giard, 1877,parasite of nemertean Lineus ruber Müller, 1774.We discovered 119 plasmodium-specific proteins, 82 of which have inferred functions based on known domains. Thirty-five of the detected proteins are orphans, at least part of which may reflect the unique evolutionary adaptations of orthonectids to parasitism. Some of the identified proteins are known effector molecules of other endoparasites suggesting convergence. Our data indicate that the plasmodium-specific proteins might be involved in the plasmodium defense against the host, host-parasite communication, feeding and nutrient uptake, growth within the host, and support of the sexual stage development. These molecular processes in orthonectids have not been described before, and the particular protein effectors remained unknown until now.

Psychosocial dysfunction and delayed sexual development among adolescents living with HIV in Lagos, Nigeria.

This was a cross-sectional study conducted at the Lagos University Teaching Hospital (LUTH), Nigeria and it involved 144 ALHIV and an equal number of HIV-negative controls who were matched for age, sex and social class. Information was obtained from participants using interviewer-administered questionnaires; their stages of sexual development and their psychosocial function were assessed using Tanner staging criteria and the Paediatric Symptom Checklist tool respectively. Data were analysed using the Statistical Package for Social Sciences software version 23. The mean (±SD) age of ALHIV and the HIV-negative controls was 14.8 (±3.0) and 14.8 (±2.9) years respectively. All the ALHIV were on HAART and 99.3% were in clinical stage 1. There was no significant difference between the prevalence of DSD among the ALHIV (9.4%) and the HIV-negative controls (6.4%) (p= 0.402). The prevalence of PSD in ALHIV and HIV-negative controls were 4.9% and 5.6% respectively (p=0.791). There was no significant association between PSD and DSD in both groups of study participants (p=0.459 and p=0.301). The prevalence of PSD and DSD were low and similar among adolescents with and without HIV, and no association was found between PSD and DSD. However, routine screening of adolescents for PSD should be practised for early identification and prompt management where indicated.

MRNA vaccines expressing malaria transmission-blocking antigens Pfs25 and Pfs230D1 induce a functional immune response

Malaria transmission-blocking vaccines (TBV) are designed to inhibit the sexual stage development of the parasite in the mosquito host and can play a significant role in achieving the goal of malaria elimination. Preclinical and clinical studies using protein–protein conjugates of leading TBV antigens Pfs25 and Pfs230 domain 1 (Pfs230D1) have demonstrated the feasibility of TBV. Nevertheless, other promising vaccine platforms for TBV remain underexplored. The recent success of mRNA vaccines revealed the potential of this technology for infectious diseases. We explored the mRNA platform for TBV development. mRNA constructs of Pfs25 and Pfs230D1 variously incorporating signal peptides (SP), GPI anchor, and Trans Membrane (TM) domain were assessed in vitro for antigen expression, and selected constructs were evaluated in mice. Only mRNA constructs with GPI anchor or TM domain that resulted in high cell surface expression of the antigens yielded strong immune responses in mice. These mRNA constructs generated higher transmission-reducing functional activity versus the corresponding alum-adjuvanted protein-protein conjugates used as comparators. Pfs25 mRNA with GPI anchor or TM maintained >99% transmission reducing activity through 126 days, the duration of the study, demonstrating the potential of mRNA platform for TBV.

Underlying Mechanisms of the Hedgehog-Like Panicle and Filamentous Leaf Tissue Symptoms Caused by Sclerospora graminicola in Foxtail Millet.

Downy mildew caused by Sclerospora graminicola is a systemic infectious disease affecting foxtail millet production in Africa and Asia. S. graminicola-infected leaves could be decomposed to a state where only the veins remain, resulting in a filamentous leaf tissue symptom. The aim of the present study was to investigate how S. graminicola influences the formation of the filamentous leaf tissue symptoms in hosts at the morphological and molecular levels. We discovered that vegetative hyphae expanded rapidly, with high biomass accumulated at the early stages of S. graminicola infection. In addition, S. graminicola could affect spikelet morphological development at the panicle branch differentiation stage to the pistil and stamen differentiation stage by interfering with hormones and nutrient metabolism in the host, resulting in hedgehog-like panicle symptoms. S. graminicola could acquire high amounts of nutrients from host tissues through secretion of β-glucosidase, endoglucanase, and pectic enzyme, and destroyed host mesophyll cells by mechanical pressure caused by rapid expansion of hyphae. At the later stages, S. graminicola could rapidly complete sexual reproduction through tryptophan, fatty acid, starch, and sucrose metabolism and subsequently produce numerous oospores. Oospore proliferation and development further damage host leaves via mechanical pressure, resulting in a large number of degraded and extinct mesophyll cells and, subsequently, malformed leaves with only veins left, that is, "filamentous leaf tissue." Our study revealed the S. graminicola expansion characteristics from its asexual to sexual development stages, and the potential mechanisms via which the destructive effects of S. graminicola on hosts occur at different growth stages.

Monilia and Monilinia : In silico genetic analysis of plant pathogenic fungi

Species of phytopathogenic fungi Monilia [J. Hill, 1751] and Monilinia [E.E. Honey, 1936] of the phylum Ascomycota have two host plant families of stone- and pome fruit trees of Rosaceae; and bushes of Ericaceae (e.g., blueberry, cranberry, heather, etc.). They infect both plants, and human (i.e., moniliasis). Due to the lack of sexual developmental stage (i.e., imperfect fungi; anamorphic fungi) the species of the order Moniliales spread by vegetative spores (i.e., conidia) and not by sexual (i.e., meiotic) ascospores. Here we report in silico data mining carried out by analyses of DNA, RNA, and protein sequences to reveal genetic distances in -and-among Monilia and Monilinia species. Sequences of the (+)ssRNA genomes of mitoviruses, which hyper parasitize the fungi cells, which parasitize the plant cells, during the ecocycles, are also analyzed. Genome size analysis of Monilinia were found to be between 30-55 x 106 bp; the cytochrome-b (cytb) genes (1543 bp of mtDNA); the transcribed cytb-mRNAs; the translated cytb proteins (381-391 aa); and the (+)ssRNA genomes of mitoviruses have showed that Botrytis cinerea (‘grey mold’) has the closest molecular similarity to Monilia and Monilinia clade (‘brown rot’). The cladogram of transcribed cytb-mRNAs (1175 nt) grouped Monilinia vaccinii-corymbosi in a distinct clade, which indicate a far genetic linkage. The application of mitovirus-infected hypovirulent phytopathogenic fungi for plant protection (i.e., virocontrol / biocontrol) is discussed.

A dual-antigen malaria vaccine targeting Pb22 and Pbg37 was able to induce robust transmission-blocking activity

BackgroundDespite years of effort to develop an effective vaccine against malaria infection, a vaccine that provides individuals with sufficient protection against malaria illness and death in endemic areas is not yet available. The development of transmission-blocking vaccines (TBVs) is a promising strategy for malaria control. A dual-antigen malaria vaccine targeting both pre- and post-fertilization antigens could effectively improve the transmission-blocking activity of vaccines against the sexual stages of the parasite.MethodsA chimeric recombinant protein Pb22-Pbg37 (Plasmodium berghei 22-P. berghei G37) composed of 19–218 amino acids (aa) of Pb22 and the N-terminal 26–88 aa of Pbg37 was designed and expressed in the Escherichia coli expression system. The antibody titers of the fusion (Pb22-Pbg37) and mixed (Pb22+Pbg37) antigens, as well as those of Pb22 and Pbg37 single antigens were evaluated by enzyme-linked immunosorbent assay. Immunofluorescence and western blot assays were performed to test the reactivity of the antisera with the native proteins in the parasite. The induction of transmission-blocking activity (TBA) by Pb22-Pbg37 and Pb22+Pbg37 were evaluated by in vitro gametocyte activation, gamete and exflagellation center formation, ookinete conversion, and in the direct mosquito feeding assay.ResultsThe Pb22-Pbg37 fusion protein was successfully expressed in vitro. Co-administration of Pb22 and Pbg37 as a fusion or mixed protein elicited comparable antibody responses in mice and resulted in responses to both antigens. Most importantly, both the mixed and fusion antigens induced antibodies with significantly higher levels of TBA than did each of the individual antigens when administered alone. In addition, the efficacy of vaccination with the Pb22-Pbg37 fusion protein was equivalent to that of vaccination with the mixed single antigens.ConclusionsDual-antigen vaccines, which expand/lengthen the period during which the transmission-blocking antibodies can act during sexual-stage development, can provide a promising higher transmission-reducing activity compared to single antigens.Graphical

The global transcriptome of Plasmodium falciparum mid-stage gametocytes (stages II-IV) appears largely conserved and gametocyte-specific gene expression patterns vary in clinical isolates.

Our overall understanding of the developmental biology of malaria parasites has been greatly enhanced by recent advances in transcriptomic analysis. However, most of these investigations rely on laboratory strains (LS) that were adapted into in vitro culture many years ago, and the transcriptomes of clinical isolates (CI) circulating in human populations have not been assessed. In this study, RNA-seq was used to compare the global transcriptome of mid-stage gametocytes derived from three short-term cultured CI, with gametocytes derived from the NF54 reference laboratory strain. The core transcriptome appeared to be consistent between CI- and LS-derived gametocyte preparations, but some important differences were also observed. A majority of gametocyte-specific genes (43/53) appear to have relatively higher expression in CI-derived gametocytes than in LS-derived gametocytes, but a K-means clustering analysis showed that genes involved in flagellum- and microtubule-based processes (movement/motility) were more abundant in both groups, albeit with some differences between them. In addition, gametocytes from one CI described as CI group II gametocytes (CI:GGII) showed gene expression variation in the form of reduced gametocyte-specific gene expression compared to the other two CI-derived gametocytes (CI gametocyte group I, CI:GGI), although the mixed developmental stages used in our study is a potential confounder, only partially mitigated by the inclusion of multiple replicates for each CI. Overall, our study suggests that there may be subtle differences in the gene expression profiles of mid-stage gametocytes from CI relative to the NF54 reference strain of Plasmodium falciparum. Thus, it is necessary to deploy gametocyte-producing clinical parasite isolates to fully understand the diversity of gene expression strategies that may occur during the sequestered development of parasite sexual stages. IMPORTANCE Maturing gametocytes of Plasmodium falciparum are known to sequester away from peripheral circulation into the bone marrow until they are mature. Blocking gametocyte sequestration can prevent malaria transmission from humans to mosquitoes, but most studies aim to understand gametocyte development utilizing long-term adapted laboratory lines instead of clinical isolates. This is a particular issue for our understanding of the sexual stages, which are known to decrease rapidly during adaptation to long-term culture, meaning that many LS are unable to produce transmissible gametocytes. Using RNA-seq, we investigated the global transcriptome of mid-stage gametocytes derived from three clinical isolates and a reference strain (NF54). This identified important differences in gene expression profiles between immature gametocytes of CI and the NF54 reference strain of P. falciparum, suggesting increased investment in gametocytogenesis in clinical isolates. Our transcriptomic data highlight the use of clinical isolates in studying the morphological, cellular features and molecular biology of gametocytes.

Plasmodium falciparum formins are essential for invasion and sexual stage development

The malaria parasite uses actin-based mechanisms throughout its lifecycle to control a range of biological processes including intracellular trafficking, gene regulation, parasite motility and invasion. In this work we assign functions to the Plasmodium falciparum formins 1 and 2 (FRM1 and FRM2) proteins in asexual and sexual blood stage development. We show that FRM1 is essential for merozoite invasion and FRM2 is required for efficient cell division. We also observed divergent functions for FRM1 and FRM2 in gametocyte development. Conditional deletion of FRM1 leads to a delay in gametocyte stage progression. We show that FRM2 controls the actin and microtubule cytoskeletons in developing gametocytes, with premature removal of the protein resulting in a loss of transmissible stage V gametocytes. Lastly, we show that targeting formin proteins with the small molecule inhibitor of formin homology domain 2 (SMIFH2) leads to a multistage block in asexual and sexual stage parasite development.

Plasmodium falciparum Development from Gametocyte to Oocyst: Insight from Functional Studies.

Malaria elimination may never succeed without the implementation of transmission-blocking strategies. The transmission of Plasmodium spp. parasites from the human host to the mosquito vector depends on circulating gametocytes in the peripheral blood of the vertebrate host. Once ingested by the mosquito during blood meals, these sexual forms undergo a series of radical morphological and metabolic changes to survive and progress from the gut to the salivary glands, where they will be waiting to be injected into the vertebrate host. The design of effective transmission-blocking strategies requires a thorough understanding of all the mechanisms that drive the development of gametocytes, gametes, sexual reproduction, and subsequent differentiation within the mosquito. The drastic changes in Plasmodium falciparum shape and function throughout its life cycle rely on the tight regulation of stage-specific gene expression. This review outlines the mechanisms involved in Plasmodium falciparum sexual stage development in both the human and mosquito vector, and zygote to oocyst differentiation. Functional studies unravel mechanisms employed by P. falciparum to orchestrate the expression of stage-specific functional products required to succeed in its complex life cycle, thus providing us with potential targets for developing new therapeutics. These mechanisms are based on studies conducted with various Plasmodium species, including predominantly P. falciparum and the rodent malaria parasites P. berghei. However, the great potential of epigenetics, genomics, transcriptomics, proteomics, and functional genetic studies to improve the understanding of malaria as a disease remains partly untapped because of limitations in studies using human malaria parasites and field isolates.

Preclinical evaluations of Pfs25-EPA and Pfs230D1-EPA in AS01 for a vaccine to reduce malaria transmission

Malaria transmission-blocking vaccine candidates Pfs25-EPA and Pfs230D1-EPA target sexual stage development of Plasmodium falciparum parasites in the mosquito host, thereby reducing mosquito infectivity. When formulated on Alhydrogel, Pfs25-EPA has demonstrated safety and immunogenicity in a phase 1 field trial, while Pfs230D1-EPA has shown superior activity to Pfs25-EPA in a phase 1US trial and has entered phase 2 field trials. Development continues to enhance immunogenicity of these candidates toward producing a vaccine to reduce malaria transmission (VRMT) with both pre-erythrocytic (i.e., anti-infection) and transmission-blocking components. GSK Adjuvant Systems have demonstrated successful potency in pre-erythrocytic vaccine trials and might offer a common platform for VRMT development. Here, we describe preclinical evaluations of Pfs25-EPA and Pfs230D1-EPA nanoparticles with GSK platforms. Formulations were stable after a series of assessments and induced superior antibody titers and functional activity in CD-1 mice, compared to Alhydrogel formulations of the same antigens.

Differential paired stage-specific expression of Babesia bovis cysteine-rich GCC2/GCC3 domain family proteins (BboGDP) during development within Rhipicephalus microplus

BackgroundBabesia bovis, an intra-erythrocytic apicomplexan parasite, is one of the causative agents of bovine babesiosis, the most important tick-borne disease of cattle in tropical and subtropical regions. Babesia bovis has a complex life-cycle that includes sexual development within the tick vector. The development of a transmission blocking vaccine to control bovine babesiosis requires the identification of antigens displayed on the surface of the parasite during its development within tick vectors. Four B. bovis cysteine-rich GCC2/GCC3 domain protein (BboGDP) family members were previously identified and are differentially expressed as discrete pairs by either blood stages or kinetes. In this study we focused on two family members, BboGDP1 and -3, that are expressed by Babesia parasites during tick infection.Methods and resultsTranscription analysis using quantitative PCR demonstrated that BboGDP1 and -3 were upregulated in in vitro-induced sexual stage parasites and during parasite development in the tick midgut. Moreover, protein expression analysis of BboGDP1 and -3 during the development of sexual stages in in vitro culture was consistent with their transcription profile. Live immunofluorescence analysis using polyclonal antibodies confirmed surface expression of BboGDP1 and -3 on in vitro-induced sexual stage parasites. In addition, fixed immunofluorescence analysis showed reactivity of anti-BboGDP1 and -3 polyclonal antibodies to kinetes.ConclusionsThe collective data indicate that BboGDP1 and -3 are expressed by kinetes and on the surface of sexual stages of the parasites. The identified parasite surface membrane proteins BboGDP1 and -3 are potential candidates for the development of a B. bovis transmission blocking vaccine.Graphical

State of sexual development in children with congenital epidermolysis bullosa

Purpose of the study. Assess the state of sexual development in children with CEB.Methods and materials. The study included 50 children aged 8.11 to 17.80 years with CEB who were treated at the National Medical Research Centre for Children’s Health (Moscow, Russia) from December 2020 to April 2022. Anthropometric parameters, the stage of sexual development on the Tanner scale, bone age, the level of pituitary (FSH, LH) and sex (estradiol, testosterone) hormones, additional hormonal parameters (cortisol, DHEA-sulfate, 17OHP, ACTH, prolactin, TSH, T3, T4, insulin), ultrasound of the pelvic organs for girls and ultrasound of the scrotum organs in boys, the results of a psychological and pedagogical examination (questionnaire according to S. Bem, ‘drawing of a person’ by K. Machover, D. Wechsler’s test) were assessed in all patients. The data are presented using median (Me), quartiles of 25 and 75% [Q25; Q75] and standard error (SE). Due to the small number of observations, nonparametric statistical criteria were used to assess the significance of the differences in the obtained indicators.Results. According to the results of the study, a statistically significant relationship was established between the occurrence of deviations in sexual development in children with CEB and the clinical form of the disease (p < 0,001). The severity of clinical manifestations of CEB according to the EBDASI scale was significantly higher in children in the group with deviations in sexual development (p = 0,000) compared with the group of children with normal sexual development. Anthropometric indicators of BAZ and HAZ in the group of children with deviations in sexual development were statistically significantly reduced (p = 0.000, p = 0.000, respectively) compared with children with normal sexual development, which means a significantly higher incidence of malnutrition in this cohort of patients. Basal levels of FSH and LH (p = 0,000, p = 0,001, respectively), estradiol and testosterone (p = 0,002, p = 0,000, respectively) were significantly lower in children with abnormalities in sexual development compared with children with normal sexual development. A statistically significant relationship was revealed in the studied groups with the stage of sexual development according to Tanner scale (p = 0,032). Among all patients with CEB (n = 50), delay puberty was diagnosed in 7 patients (14%), while all patients suffered from RDEB and had low basal LH levels, reduced estradiol and testosterone values, which allowed them to be diagnosed with hypogonadotropic hypogonadism (HH).Conclusions. In patients with CEB with the most severe degree of clinical manifestations, accompanied by decompensated malnutrition of multifactorial genesis, there are deviations in sexual development and upon reaching the age of 13 years, girls and boys develop transient (symptomatic).

Sexual Maturity Scale of the Swimming Crab Callinectes pallidus (Rochebrune, 1883) from Lake Nokoué in South Benin, West Africa

Aims: Reproductive parameters are important tools for the sustainable management of halieutic species in the water bodies of its exploitation. This study intends to characterize the sexual maturity scale of swimming crab Callinectes pallidus, perform testicular and ovarian structures in order to develop a specific maturity scale.
 Study Design: This study was carried out with macroscopic and microscopic observations of 394 specimens of crabs including 206 females and 188 males.
 Place and Duration of Study: The specimens used in this study were collected monthly from March to July 2018 from small scale fisheries of Lake Nokoué (Benin).
 Methodology: At the laboratory, each crab specimen is identified and its sex determined. Grids for describing each sexual maturity stage of the crab C. pallidus were developed based on the Zairon et al. (2015) scale. Histological sections of female and male gonads were analyzed in order to certify the different stages of sexual development noted from the macroscopic study.
 Results: Macroscopic analysis of external and internal anatomy and histological study of male and female gonads of C. pallidus allowed to establish a sexual maturity scale with 5 stages in females and 4 stages in males respectively. Macroscopic observations made with a binocular magnifying glass allowed to describe the characteristic features of the sexual development cycle of the species. The histological study confirmed the results of the macroscopic analysis. Significant difference was recorded between the mean oocytes diameters of the different oocyte development stages (p<0.05).
 Conclusion: The outcomes of this constitute an important database for the assessment of the reproductive parameters of the species in the water bodies of its exploitation. The stages IV and V are the mature steps in the female development whereas the stages III and IV are that of the male. It is recommended to allow C. pallidus reach the sexual maturity size (5,95 cm for female and 6,38 cm for male) before their exploitation to ensure the sustainable management of the species.

Quality Evaluation of Gilts and Sow’s Oocytes During In Vitro Maturation Based on Mitochondria Distribution


 Mitochondria is the most frequently studied cell organelle in relation to development competence of the oocytes both in vivo and in vitro. Developmental competence of oocyte is crucial for embryonic development, the molecular activity until the activation of the embryonic genome being based on the maternal reserves stored in the oocyte. Study was designed to qualitatively evaluate 133 COC class C1 and class C2 gilts and sow’s oocytes according to the distribution of mitochondria after IVM, based on Rhodamine 123 stain solution. Regarding mitochondrial distribution, the percentage of oocytes that had a homogeneous distribution around the germinal vesicle was higher in group C1 than C2 and also higher in sow oocytes than in gilts oocytes. For class 1, the difference was 10.58% in favor of sows, the same trend is maintained for C2, the difference being 4.37%. The results are confirmed by morphological examination, where C1 sow’s oocytes matured 26.09% more, compared to gilts oocytes, a difference maintained for C2 oocytes, being 27.4% more sow’s oocytes IVM compared to gilts. Based on mitochondrial distribution we observed that the stage of sexual development of females influences the IVM of oocytes. For pig’s IVF it’s recommended to use C1 sow oocytes.

Inhibition of sexual stage-specific proteins results in reduced numbers of sexual stages and oocysts of Cystoisospora suis (Apicomplexa: Coccidia) in vitro

Parasites of the order Coccidia (phylum: Alveolata, subphylum: Apicomplexa) have sophisticated life cycles that include a switch from asexual to sexual development, characterised by distinct cell types. During the development of gametes (gamogony), substantial changes occur at the cellular and subcellular levels, leading to cell fusion of micro- and microgametes, and the development of a zygote that forms a protective outer layer for environmental survival as an oocyst, the transmissible stage. Studies on the porcine coccidian Cystoisospora suis already identified changes in transcription profiles during different time points in the parasite’s development and identified proteins with potential roles in the sexual development of this parasite. Here, we focus on three proteins that are possibly involved in the sexual development of C. suis. Enkurin and hapless protein 2 (HAP2) play important roles in signal transduction and gamete fusion during the fertilisation process, and oocyst wall forming protein 1 (OWP1) is a homologue of oocyst wall forming proteins of related parasites. We evaluated their locations in the different life cycle stages of C. suis and their inhibition by specific antibodies in vitro. Immunolocalization detected enkurin in merozoites and sporulated oocysts, HAP2 in merozoites and microgamonts, and OWP2 in merozoites, macrogamonts, oocysts and sporozoites. Up to 100% inhibition of the development of sexual stages and oocyst formation with purified chicken immunoglobulin IgY sera against recombinant enkurin, HAP2, and especially OWP1, were demonstrated. We conclude that the three investigated sexual stage-specific proteins constitute targets for in vivo intervention strategies to interrupt parasite development and transmission to susceptible hosts.