Deleterious genetic changes in AGTPBP1 result in teratozoospermia with sperm head and flagella defects.

Approximately 10%-15% of couples worldwide are infertile, and male factors account for approximately half of these cases. Teratozoospermia is a major cause of male infertility. Although various mutations have been identified in teratozoospermia, these can vary among ethnic groups. In this study, we performed whole-exome sequencing to identify genetic changes potentially causative of teratozoospermia. Out of seven genes identified, one, ATP/GTP Binding Protein 1 (AGTPBP1), was characterized, and three missense changes were identified in two patients (Affected A: p.Glu423Asp and p.Pro631Leu; Affected B: p.Arg811His). In those two cases, severe sperm head and tail defects were observed. Moreover, AGTPBP1 localization showed a fragmented pattern compared to control participants, with specific localization in the neck and annulus regions. Using murine models, we found that AGTPBP1 is localized in the manchette structure, which is essential for sperm structure formation. Additionally, in Agtpbp1-null mice, we observed sperm head and tail defects similar to those in sperm from AGTPBP1-mutated cases, along with abnormal polyglutamylation tubulin and decreasing △-2 tubulin levels. In this study, we established a link between genetic changes in AGTPBP1 and human teratozoospermia for the first time and identified the role of AGTPBP1 in deglutamination, which is crucial for sperm formation.

A Comprehensive Genetic Study of Microtubule-Associated Gene Clusters for Male Infertility in a Taiwanese Cohort.

Advanced reproductive technologies are utilized to identify the genetic mutations that lead to spermatogenic impairment, and allow informed genetic counseling to patients to prevent the transmission of genetic defects to offspring. The purpose of this study was to identify potential single nucleotide polymorphisms (SNPs) associated with male infertility. Genetic variants that may cause infertility are identified by combining the targeted next-generation sequencing (NGS) panel and whole exome sequencing (WES). The validation step of Sanger sequencing adds confidence to the identified variants. Our analysis revealed five distinct affected genes covering seven SNPs based on the targeted NGS panel and WES data: SPATA16 (rs16846616, 1515442, 1515441), CFTR (rs213950), KIF6 (rs2273063), STPG2 (r2903150), and DRC7 (rs3809611). Infertile men have a higher mutation rate than fertile men, especially those with azoospermia. These findings strongly support the hypothesis that the dysfunction of microtubule-related and spermatogenesis-specific genes contributes to idiopathic male infertility. The SPATA16, CFTR, KIF6, STPG2, and DRC7 mutations are associated with male infertility, specifically azoospermia, and a further examination of this genetic function is required.

Localization Patterns of RAB3C Are Associated with Murine and Human Sperm Formation.

Background and Objectives: Septins (SEPTs) are highly conserved GTP-binding proteins and the fourth component of the cytoskeleton. Polymerization of SEPTs contributes to several critical cellular processes such as cytokinesis, cytoskeletal remodeling, and vesicle transportation. In our previous study, we found that SEPT14 mutations resulted in teratozoospermia with >87% sperm morphological defects. SEPT14 interactors were also identified through proteomic assays, and one of the peptides was mapped to RAB3B and RAB3C. Most studies on the RAB3 family have focused on RAB3A, which regulates the exocytosis of neurotransmitters and acrosome reactions. However, the general expression and patterns of the RAB3 family members during human spermatogenesis, and the association between RAB3 and teratozoospermia owing to a SEPT14 mutation, are largely unknown. Materials and Methods: Human sperm and murine male germ cells were collected in this study and immunofluorescence analysis was applied on the collected sperm. Results: In this study, we observed that the RAB3C transcripts were more abundant than those of RAB3A, 3B, and 3D in human testicular tissues. During human spermatogenesis, the RAB3C protein is mainly enriched in elongated spermatids, and RAB3B is undetectable. In mature human spermatozoa, RAB3C is concentrated in the postacrosomal region, neck, and midpiece. The RAB3C signals were delocalized within human spermatozoa harboring the SEPT14 mutation, and the decreased signals were accompanied by a defective head and tail, compared with the healthy controls. To determine whether RAB3C is involved in the morphological formation of the head and tail of the sperm, we separated murine testicular tissue and isolated elongated spermatids for further study. We found that RAB3C is particularly expressed in the manchette structure, which assists sperm head shaping at the spermatid head, and is also localized at the sperm tail. Conclusions: Based on these results, we suggest that the localization of RAB3C proteins in murine and human sperm is associated with SEPT14 mutation-induced morphological defects in sperm.

ACTN4 Mediates SEPT14 Mutation-Induced Sperm Head Defects.

Septins (SEPTs) are highly conserved GTP-binding proteins and the fourth component of the cytoskeleton. Polymerized SEPTs participate in the modulation of various cellular processes, such as cytokinesis, cell polarity, and membrane dynamics, through their interactions with microtubules, actin, and other cellular components. The main objective of this study was to dissect the molecular pathological mechanism of SEPT14 mutation-induced sperm head defects. To identify SEPT14 interactors, co-immunoprecipitation (co-IP) and nano-liquid chromatography-mass spectrometry/mass spectrometry were applied. Immunostaining showed that SEPT14 was significantly localized to the manchette structure. The SEPT14 interactors were identified and classified as (1) SEPT-, (2) microtubule-, (3) actin-, and (4) sperm structure-related proteins. One interactor, ACTN4, an actin-holding protein, was selected for further study. Co-IP experiments showed that SEPT14 interacts with ACTN4 in a male germ cell line. SEPT14 also co-localized with ACTN4 in the perinuclear and manchette regions of the sperm head in early elongating spermatids. In the cell model, mutated SEPT14 disturbed the localization pattern of ACTN4. In a clinical aspect, sperm with mutant SEPT14, SEPT14A123T (p.Ala123Thr), and SEPT14I333T (p.Ile333Thr), have mislocalized and fragmented ACTN4 signals. Sperm head defects in donors with SEPT14 mutations are caused by disruption of the functions of ACTN4 and actin during sperm head formation.

Feasibility and Safety of Absorbable Knotless Wound Closure Device in Laparoscopic Myomectomy.

Purpose. Myomectomy has been performed through laparoscopy. Suturing is known as rate-limiting step in laparoscopic myomectomy. The present study was aimed at comparing the clinical outcomes of absorbable knotless wound closure device with the results of conventional suturing. Methods. This prospective study included 62 women who underwent laparoscopic myomectomy at Taipei City Hospital, Zhongxiao Branch, from January 2010 through to August 2012. The patients were randomized into two groups according to suturing materials, the knotless group and the 2-0 Vicryl suture group. Patient demographics, overall operative time, and intraoperative blood loss were compared between two groups. Results. Demographic characteristics and laboratory variables before surgery were comparable. Operative time was significantly shorter in knotless group compared with that in 2-0 Vicryl suture group (112 ± 47 versus 147 ± 63 minutes; p < 0.05). The results revealed a significant difference in intraoperative blood loss between two groups (knotless versus 2-0 Vicryl: 112.8 ± 54.2 versus 143.6 ± 64.9). Use of absorbable knotless wound closure device was associated with greater hemostasis compared with that of 2-0 Vicryl. During a 2-year follow-up period, 12 patients (46.2%) from the group with absorbable knotless wound closure device and 14 patients (38.9%) from 2-0 Vicryl suture group became pregnant. Conclusion. Closure of myometrium using absorbable knotless wound closure device after laparoscopic myomectomy resulted in a shorter operative time and less blood loss.

Differential spermatozoal protein expression profiles in men with varicocele compared to control subjects: upregulation of heat shock proteins 70 and 90 in varicocele.

OBJECTIVE: To identify candidate proteins with potential roles in varicocele. METHODS: This case-control study recruited 20 patients with varicocele (grade II) and 20 age-matched healthy control subjects. Two-dimensional gel electrophoreses (2-DE) and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) were used to screen for and identify differentially expressed proteins in sperm samples from the 40 study subjects. RESULTS: A total of 15 proteins were repeatedly detected with significant differences in percent spot volume (P <.05) and were identified as proteins of interest. Of these, 4 showed significantly lower expression in the varicocele group compared to the control group, and 11 showed significantly higher expression in the varicocele group, including heat shock proteins (HSPs) 70 and 90. Significant upregulation of HSP 70 and HSP 90 in subjects with varicocele was further validated by Western blot and immunocytochemistry. CONCLUSION: These results suggest that increased expression of HSP 70 and HSP 90 may be associated with infertility in varicocele. Delineation of a potential subset of heat-related proteins specifically regulated in the context of varicocele may allow for the detailed elucidation of the mechanism underlying infertility in this condition.

Motility and protein phosphorylation in healthy and asthenozoospermic sperm.

The majority of male infertility results from poor sperm motility. A direct link between altered protein phosphorylation and aberrant sperm motility has not been established. To address this issue, sperm samples obtained from 20 donors with healthy sperm and 20 donors with aberrantly motile sperm were subjected to computer assisted semen analysis (CASA), proteomic analysis, Western blot, and immunofluorescent staining. Proteomic analysis identified 12 protein spots as having differential phosphorylation, including gamma-tubulin complex associated protein 2 (GCP2). Western blot and immunofluorescence demonstrated differential expression of gamma-tubulin between healthy and aberrantly motile sperm. In conclusion, hypophosphorylated proteins and reduced expression of gamma-tubulin may be associated with low motility sperm.

Isolated torsion of the fallopian tube in a 14-year-old adolescent.

OBJECTIVE: Torsion of adnexa is relatively common, but isolated torsion of the fallopian tube is rare. It should be considered in all adolescents who present with acute pelvic pain. Laparoscopy or laparotomy is often necessary to establish the diagnosis. This report focuses on a 14-year-old girl with isolated tubal torsion who presented with acute pelvic pain. CASE REPORT: A 14-year-old adolescent was admitted to our hospital because of acute right-sided abdominal pain without vomiting and diarrhea. Pelvic ultrasound showed an adnexal mass. Conservative treatment was given but did not improve her condition. Emergent laparoscopy was performed due to persistent symptoms, which later confirmed the diagnosis of isolated torsion of the fallopian tube. Pathology showed hemosalpinx with necrosis. CONCLUSION: Isolated torsion of the fallopian tube is an uncommon event, especially in adolescents. It must be kept in mind whenever a young girl presents with low abdominal pain and pelvic mass on ultrasound. Prompt laparoscopic intervention may allow for early diagnosis, treatment and preservation of the tube if possible.

Predicted value of renin activity in a woman who had severe ovarian hyperstimulation syndrome with internal jugular vein thrombosis.

OBJECTIVE: To assess plasma renin activity in a patient with severe ovarian hyperstimulation syndrome (OHSS) and internal jugular vein thrombosis. DESIGN: Case report. SETTING: University-affiliated infertility center. PATIENT(S): A 33-year-old woman with OHSS and internal jugular vein thrombosis. INTERVENTION(S): Controlled ovulation hyperstimulation with recombinant FSH induction. MAIN OUTCOME MEASURE(S): Plasma renin activity (PRA), color Doppler ultrasound of the neck. RESULT(S): The patient had internal jugular vein thrombosis caused by severe OHSS. The PRA was significantly elevated during the acute stage and subsequently declined after resolution of the OHSS. CONCLUSION(S): In this patient elevated PRA appeared to be associated with the development of OHSS and thrombosis. The implication of the ovarian renin-angiotensin system in the development of OHSS and thrombosis is relevant.

Continuous abdominal paracentesis for management of late type severe ovarian hyperstimulation syndrome.

The ovarian hyperstimulation syndrome (OHSS) is often observed in patients undergoing assisted reproductive technology (ART). In severe form OHSS is a serious and potentially life-threatening. Here we report a 36-year-old woman with primary infertility due to endometriosis who underwent controlled ovarian hyperstimulation. Ten days later, severe late-onset ovarian hyperstimulation syndrome, severe ascites and pulmonary effusion, developed. Continuous abdominal paracentesis of 5000 mL/day was performed on the third day. With this procedure, ascitic fluid was drained efficiently and the patient's condition improved. This report suggests that early continuous abdominal paracentesis with drainage of ascitic fluid is an efficacious procedure for management of the severe ovarian hyperstimulation syndrome as soon as euvolemia is achieved clinically.