Fertility outcomes in microdissection testicular sperm extraction for non-obstructive azoospermia

Background: The objective of this study was to evaluate outcome of micro dissection testicular sperm extraction (micro TESE) and intracytoplasmic sperm injection (ICSI) for treatment of non-obstructive azoospermia (NOA). Methods: We retrospectively analysed data of 96 consecutive patients with clinical NOA who were treated with micro TESE by single surgeon, between January 2022 and December 2022, in Lifeline superspeciality hospital Adoor, Kerela. Embryological and clinical outcomes were demonstrated based on ICSI-IVF cycles using fresh or frozen sperms, different etiologies of NOA and various counts of sperms retrieved. Results: 96 men underwent micro TESE and 72.9% (70/96) of them had sperms retrieved. ICSI performed in 64 couples. Of those, 41 reached the stage of embryo transfer (ET). Of the couples who underwent embryo transferred, 18 (43%) resulted in biochemical pregnancies and 7 (17%) clinical pregnancies. There was a significant difference in the testicular volume and serum FSH levels between micro-TESE positive and negative groups (p=0.000). Retrieval rates were higher in group of men with normal testicular volume and FSH<12. Clinical pregnancy rate was around 11% in couples who had sperms retrieved by micro TESE. The sperm retrieval rates were higher in men with age <40 years. Similarly younger the female age more was the pregnancy rate. Out of various etiologies idiopathic NOA and Klinefelter syndrome had better sperm retrieval rate in our study which was statistically significant. Total fertilization rate and blasts rates were 79.7% and 51.6% out of ICSI. Conclusions: Microdissection testicular sperm extraction is an effective treatment for NOA with higher rate of sperm retrieval and pregnancy rate. The increasing success rates over several years indicate the importance of surgical skill and laboratory staff experience

Mix-and-Match System for the Enzymatic Synthesis of Enantiopure Glycerol-3-Phosphate-Containing Capsule Polymer Backbones from Actinobacillus pleuropneumoniae, Neisseria meningitidis, and Bibersteinia trehalosi

Capsule polymers are crucial virulence factors of pathogenic bacteria and are used as antigens in glycoconjugate vaccine formulations. Some Gram-negative pathogens express poly(glycosylglycerol phosphate) capsule polymers that resemble Gram-positive wall teichoic acids and are synthesized by TagF-like capsule polymerases. So far, the biotechnological use of these enzymes for vaccine developmental studies was restricted by the unavailability of enantiopure CDP-glycerol, one of the donor substrates required for polymer assembly. Here, we use CTP:glycerol-phosphate cytidylyltransferases (GCTs) and TagF-like polymerases to synthesize the poly(glycosylglycerol phosphate) capsule polymer backbones of the porcine pathogen Actinobacillus pleuropneumoniae, serotypes 3 and 7 (App3 and App7). GCT activity was confirmed by high-performance liquid chromatography, and polymers were analyzed using comprehensive nuclear magnetic resonance studies. Solid-phase synthesis protocols were established to allow potential scale-up of polymer production. In addition, one-pot reactions exploiting glycerol-kinase allowed us to start the reaction from inexpensive, widely available substrates. Finally, this study highlights that multidomain TagF-like polymerases can be transformed by mutagenesis of active site residues into single-action transferases, which in turn can act in trans to build-up structurally new polymers. Overall, our protocols provide enantiopure, nature-identical capsule polymer backbones from App2, App3, App7, App9, and App11, Neisseria meningitidis serogroup H, and Bibersteinia trehalosi serotypes T3 and T15

Prediction of Human Disease Genes by Human-Mouse Conserved Coexpression Analysis

One of the most limiting aspects of biological research in the post-genomic era is the capability to integrate massive datasets on gene structure and function for producing useful biological knowledge. In this report we have applied an integrative approach to address the problem of identifying likely candidate genes within loci associated with human genetic diseases. Despite the recent progress in sequencing technologies, approaching this problem from an experimental perspective still represents a very demanding task, because the critical region may typically contain hundreds of positional candidates. We found that by concentrating only on genes sharing similar expression profiles in both human and mouse, massive microarray datasets can be used to reliably identify disease-relevant relationships among genes. Moreover, we found that integrating the coexpression criterion with systematic phenome analysis allows efficient identification of disease genes in large genomic regions. Using this approach on 850 OMIM loci characterized by unknown molecular basis, we propose high-probability candidates for 81 genetic diseases