Novel PATL2 variants cause female infertility with oocyte maturation defect.

PURPOSE: Oocyte maturation defect (OOMD) is a rare cause of in vitro fertilization failure characterized by the production of immature oocytes. Compound heterozygous or homozygous PATL2 mutations have been associated with oocyte arrest at the germinal vesicle (GV), metaphase I (MI), and metaphase II (MII) stages, as well as morphological changes. METHODS: In this study, we recruited three OOMD cases and conducted a comprehensive multiplatform laboratory investigation. RESULTS: Whole exome sequence (WES) revealed four diagnostic variants in PATL2, nonsense mutation c.709C > T (p.R237*) and frameshift mutation c.1486_1487delinsT (p.A496Sfs*4) were novel mutations that have not been reported previously. Furthermore, the pathogenicity of these variants was predicted using in silico analysis, which indicated detrimental effects. Molecular dynamic analysis suggested that the A496S variant disrupted the hydrophobic segment, leading to structural changes that affected the overall protein folding and stability. Additionally, biochemical and molecular experiments were conducted on cells transfected with wild-type (WT) or mutant PATL2 (p.R237* and p.A496Sfs*4) plasmid vectors. CONCLUSIONS: The results demonstrated that PATL2A496Sfs*4 and PATL2R237* had impacts on protein size and expression level. Interestingly, expression levels of specific genes involved in oocyte maturation and early embryonic development were found to be simultaneously deregulated. The findings in our study expand the variation spectrum of the PATL2 gene, provide solid evidence for counseling on future pregnancies in affected families, strongly support the application of in the diagnosis of OOMD, and contribute to the understanding of PATL2 function.

Biomechanical study of extramedullary and intramedullary fixation in the treatment of unstable intertrochanteric reversed-tilt fractures of the femur.

Background: To investigate the efficacy of the 135° hip screw, 95° intramedullary hip screw (IMHS) and 95° hip screw in the treatment of intertrochanteric reverse dip fracture of the femur. Methods: We retrospectively analyzed 125 matched pairs of human femurs (median age 64 years) which were osteotomized at a 33° angle in the left femur and extended downward from the minor trochanter to simulate a reverse oblique intertrochanteric fracture. The right femur served as a control. The left femur (n=4) was implanted with a 135° hip screw, 95° hip screw, or IMHS. A strain detector was placed distal to the fracture site to monitor fragment strain. The lateral displacement of the proximal femur was measured by a linear variable differential transformer. An Instron tester measured stiffness, strain, and lateral displacement at 25° adduction, and 90° adduction with vertical loads on the femoral head. A 2 cm gap was then formed at the fracture site to simulate comminution and the mechanical test was repeated. Results: Before the formation of the gap, there was no significant difference in stiffness among different bone structures (P>0.05), but after the formation of the gap, the stiffness of all the adduction structures decreased (P=0.03), and the difference in adduction was statistically significant (135° hip screw: 46.6%±3%; 95° hip screw: 22.9%±2%; IMHS: 53.7%±7.8%; P<0.05). Similar results were found for the abduction and buckling positions. There was no significant difference in the lateral displacement of the gap before (P=0.92) and after (P=0.26), but a significant difference in the failure load was found (135° hip screw: 1,222±560 N; 95° hip screw: 2,566±283 N; IMHS: 4,644±518 N; P=0.02). Conclusions: There was no statistically significant difference in stiffness among different structures (P>0.05). However, in the presence of gaps, IMHS bone implant structures are much stiffer than 135° and 95° structures and have a greater destructive load.

Novel Biallelic DNAH1 Variations Cause Multiple Morphological Abnormalities of the Sperm Flagella.

Sperm motility is vital to human reproduction, and malformed sperm flagella can cause male infertility. Individuals with multiple morphological abnormalities of the flagella mostly have absent, short, coiled, bent, and/or irregular-caliber flagella. In this study, a patient with male infertility underwent a physical examination along with his wife. Genetic testing was performed by whole-exome sequencing of the couple, and Sanger sequencing was performed for validation. Novel biallelic variations in the DNAH1: (NM_015512.4) gene consisting of c.1336G>C (p.E446Q) and c.2912G>A (p.R971H) were identified. In silico structural analysis revealed that the amino acid residues affected by the variation were evolutionarily conserved, and the variant p.R971H influenced the stability of the DNAH1 protein. Morphological studies of the patient's sperm showed defects in its flagella. Results of Papanicolaou staining and scanning electron microscopy demonstrated coiled and short flagella with multiple anomalies. Transmission electron microscopy of the sperm flagella showed that the inner dynein arm and radial spoke were absent, and the dense fiber and microtubule doublets were displaced. Quantitative PCR of the mRNA of the patient's sperm showed that the expression of DNALI1 was dramatically reduced. Collectively, these findings elucidated the genetic cause of the family's infertility and provided insight into the functioning of the DNAH1 gene.

Effects of dissolved oxygen on the sludge dewaterability and extracellular polymeric substances distribution by bioleaching.

Bioleaching is a biological conditioning technology for sludge, which not only improves sludge dewatering performance but also removes heavy metals from sludge. As the bioleaching process is a comprehensive biological and chemical process, it is necessary to explore the effects of dissolved oxygen (DO) concentrations on bioleaching efficiency. Three bioleaching experiments with different DO concentrations (T1: 0.8-3.1 mg/L, T2: 3.1-5.5 mg/L, T3: 5.5-7.5 mg/L) were conducted for five days. The sludge dewatering efficiency was evaluated using capillary suction time (CST) and specific resistance to filtration (SRF). The relationship between sludge dewaterability and extracellular polymeric substance (EPS) fraction distribution was investigated. In the treatment with the highest DO concentration, the minimum values of SRF and CST were 4.31 × 1011 m/kg and 13.5 s, which occurred earlier than the treatment with the lower DO concentrations by approximately 24-48 h. A significant decrease (83.4-93.2%) in tightly bound EPS (TB-EPS) protein (PN) was observed in all treatments, while a positive correlation (r = 0.924, P < 0.01) was observed between SRF and PN content in TB-EPS. A relatively higher abundance of Acidithiobacillus was found with the increase in DO concentration. Additionally, other genera including Metallibacterium, Alicyclobacillus, Acidibacter, Acidocella, and Luteococcus also played important roles in EPS biodegradation. These results revealed that increasing the DO concentration could improve sludge dewatering performance and heavy metal removal by enhancing bioleaching microbial activity, the degradation of PN in TB-EPS, and sludge floc fragmentation, but only if sufficient energy sources were provided.

Letrozole ovulation induction: an effective option in endometrial preparation for frozen-thawed embryo transfer.

PURPOSE: To evaluate the clinical efficacy of letrozole on ovulation induction and hormone replacement therapy (HRT) during endometrial preparation for frozen-thawed embryo transfer (FET). METHODS: We analyzed totally 1,230 cycles of patients that underwent FET from October 2010 to September 2012. Seven hundred and thirteen cycles of patients with ovulation disorders that underwent FET were randomly assigned to two groups by case control study. 359 cycles received letrozole ovulation induction and 354 cycles received HRT during endometrial preparation for FET, respectively. In the corresponding period, 517 cycles of patients with normal ovulation in the natural cycle group for FET endometrial preparation served as controls. Reproduction-related clinical outcomes of patients in the three groups were compared. RESULTS: The embryo implantation rate of patients in letrozole group (30.4 %) was significantly higher than the HRT group (22.8 %, P < 0.05). The clinical pregnancy rate of patients in the letrozole group (53.2 %) was significantly higher than the HRT group (44.4 %, P < 0.05), while no significant difference was observed between the letrozole and natural cycle groups (51.3 %, P > 0.05). Estradiol levels on the day of human chorionic gonadotropin administration in the letrozole group were significantly lower than those in the natural cycle group (280.32 ± 125.39 pg/ml and 351.06 ± 123.03 pg/ml, respectively; P < 0.05). The live birth rate of patients in letrozole group (44.6 %) was significantly higher than the HRT group (32.5 %, P < 0.05), while abortion rate (12.0 %) was significantly lower than the HRT group (21.0 %, P < 0.05). There were no significant differences in number of mature follicles, endometrial thickness, duration of follicle growth between the letrozole and the natural cycle groups, and there were no significant differences in twin birth rate and ectopic pregnancy rate among the three groups (all P values >0.05). CONCLUSIONS: Ovulation induction with letrozole during endometrial preparation for FET has a higher rate of pregnancy success and a lower abortion rate than HRT. Letrozole treatment exhibits clinical progression and outcomes similar to those patients undergoing a natural cycle or normal ovulation cycle. Therefore, letrozole treatment may be an effective option in endometrial preparation for FET in patients with ovulation disorders or irregular menstruation.