The IMSI procedure improves poor embryo development in the same infertile couples with poor semen quality: A comparative prospective randomized study

<p>Abstract</p> <p>Background</p> <p>Sperm of poor quality can negatively affect embryo development to the blastocyst stage. The aim of this comparative prospective randomized study was to evaluate the role of an intracytoplasmic morphologically selected sperm injection (IMSI) in the same infertile couples included in the programme of intracytoplasmic sperm injection (ICSI) due to their indications of male infertility which had resulted in all arrested embryos following a prolonged 5-day culture in previous ICSI cycles.</p> <p>Methods</p> <p>Couples exhibiting poor semen quality and with all arrested embryos following a prolonged 5-day culture in previous ICSI cycles were divided into two groups: <it>Group 1: IMSI group </it>(n = 20) with IMSI performed in a current attempt and <it>Group 2: ICSI group </it>(n = 37) with a conventional ICSI procedure performed in a current attempt of <it>in vitro </it>fertilization. Fertilization rate, embryo development, implantation, pregnancy and abortion rates were compared between current IMSI and conventional ICSI procedures, and with previous ICSI attempts.</p> <p>Results</p> <p>The <it>IMSI group </it>was characterized by a higher number of blastocysts per cycle than the <it>ICSI group </it>(0.80 vs. 0.65) after a prolonged 5-day embryo culture. There was a significantly lower number of cycles with all arrested embryos and cycles with no embryo transfer in the <it>IMSI group </it>versus the <it>ICSI group </it>(0% vs. 27.0%, p = 0.048). After the transfer of embryos at the blastocyst or morula stage (on luteal day 5) a tendency toward higher implantation and pregnancy rates per cycle was achieved in the <it>IMSI group </it>compared to the <it>ICSI group </it>(17.1% vs. 6.8%; 25.0% vs. 8.1%, respectively), although not statistically significant. After IMSI, all pregnancies achieved by the blastocyst transfer were normally on-going, whereas after ICSI, two of three pregnancies ended in spontaneous abortion. After IMSI, two pregnancies were also achieved by the morula stage embryos, whereas after the conventional ICSI procedure, embryos at the morula stage did not implant.</p> <p>Conclusions</p> <p>The IMSI procedure improved embryo development and the laboratory and clinical outcomes of sperm microinjection in the same infertile couples with male infertility and poor embryo development over the previous ICSI attempts.</p

Transcriptomic signatures for human male infertility

Introduction: Male infertility is a common, complex disorder. A better understanding of pathogenesis and etiology is needed for timely diagnosis and treatment. The aim of this study, therefore, was to identify genes involved in the pathogenesis of idiopathic male infertility based on data from transcriptomic level supported with data from genomic level.Materials and methods: First, we performed whole gene expression analysis in 20 testis biopsy samples of patients with severely impaired (10) and normal spermatogenesis (10). Further, we have performed systematic review of comparable male infertility studies and overlapped the most significantly expressed genes identified in our study with the most differentially expressed genes from selected studies. Gene Ontology analysis and KEGG functional enrichment have been performed with Enrichr analysis tool. Additionally, we have overlapped these genes with the genes where rare variants have been identified previously.Results: In 10 patients with severely impaired spermatogenesis and 10 controls, we identified more than 1,800 differentially expressed genes (p &lt; 0.001). With the systematic review of three previously performed microarray studies that have met inclusion criteria we identified 257 overlapped differentialy expressed genes (144 downregulated and 113 upregulated). Intersection of genes from transcriptomic studies with genes with identified rare variants revealed a total of 7 genes linked with male infertility phenotype (CYP11A1, CYP17A1, RSPH3, TSGA10, AKAP4, CCIN, NDNF).Conclusion: Our comprehensive study highlighted the role of four genes in pathogenesis of male infertility and provided supporting evidence for three promising candidate genes which dysfunction may result in a male infertility disorder

Mammal responses to global changes in human activity vary by trophic group and landscape

Wildlife must adapt to human presence to survive in the Anthropocene, so it is critical to understand species responses to humans in different contexts. We used camera trapping as a lens to view mammal responses to changes in human activity during the COVID-19 pandemic. Across 163 species sampled in 102 projects around the world, changes in the amount and timing of animal activity varied widely. Under higher human activity, mammals were less active in undeveloped areas but unexpectedly more active in developed areas while exhibiting greater nocturnality. Carnivores were most sensitive, showing the strongest decreases in activity and greatest increases in nocturnality. Wildlife managers must consider how habituation and uneven sensitivity across species may cause fundamental differences in human–wildlife interactions along gradients of human influence.Peer reviewe

Adapted methods for scanning electron microscopy (SEM) in assessment of human sperm morphology

Infertility is a widespread problem, and in some cases, the routine basic semen analysis is not sufficient to detect the cause of male infertility. The use of the scanning electron microscope (SEM) could provide a detailed insight into spermatozoa morphology, but it requires specific sample preparation techniques. The purpose of this study was to select, adjust, and optimize a method for the preparation of spermatozoa samples prior to SEM analysis, and to establish the protocol required for its use in clinical practice. We examined sperm samples of 50 men. The samples were fixed with modified iso-osmolar aldehyde solution followed by osmium post-fixation. In the first method, dehydration of the cells and subsequent critical point drying (CPD) were performed on a coverslip. In the second method, the samples were dehydrated in centrifuge tubes; hexamethyldisilazane (HMDS) was used as a drying agent instead of CPD, and the samples were air-dried. The third procedure was based on a membrane filter. The samples were dehydrated and dried with HMDS in a Gooch crucible, continuously, without centrifugation or redispersion of the sample. Our results showed that the fixation with modified iso-osmolar aldehyde solution followed by osmium post-fixation, and combined with dehydration and CPD on a coverslip, is the most convenient procedure for SEM sample preparation. In the case of small-size samples or low sperm concentration, dehydration and drying with HMDS on the membrane filter enabled the best reliability, repeatability, and comparability of the results. The presented procedures are suitable for routine use, and they can be applied to confirm as well as to correct a diagnosis

PSYCHOSOCIAL CHARACTERISTICS OF FAMILIES WITH ICSI CHILDREN AND THESE CHILDREN’S DEVELOPMENT

Background. The aim of the study was to find whether children conceived through ICSI and their families differ from the children conceived in normally fertile families. Methods. The study group (SG) consisted of 41 children aged 3 years conceived through ICSI: 22 boys, 19 girls, 6 pairs of twins, and the control group (CG) of 41 children matched for sex, age and twin pairs. Family characteristics were assessed using a self-administered questionnaire on family characteristics, conception, pregnancy, labour and delivery, and postpartum period. The children’s development was assessed using the Developmental Čuturić Scale, and the parents filled in the questionnaire from SPP-3 on signs of inadequate child’s adaptation. The mother’s personality was assessed using the BFQ. Differences between the groups were analyzed using SPSS for Windows. Results. Comparison between the SG and CG showed the following statistically significant differences: in the SG mothers were older, the ICSI child was more frequently the only child, the mean duration of infertility treatment was 4 years, in most couples pregnancy occurred by the 3rd ICSI attempt, the family had a better socio-economic status. During the pregnancy, the SG mothers were less ambivalent towards pregnancy and the relationship between the partners improved, after delivery their psychic condition deteriorated, they were more concerned whether their child would develop normally. In the SG parents’ opinion, the quality of life improved after their baby’s birth; they described their children as more demanding and more restless. More ICSI children were cared after by their grand parents than their CG peers that mostly attended kindergarten. The development of all children was normal. Although the sum of signs of inadequate adaptation was comparable between the groups, the ICSI children demonstrated stronger intensity of individual sings. Factor analysis showed very heterogenic latent structure with many components that explained low percentage of variance; there was a clear connection between child’s signs of inadequate adaptation and maternal personality characteristics. The SG mothers achieved on BFQ a statistically lower score in agreeableness (P = 0.025) and openness (P = 0.008), and a higher score in the unsincerity scale (P = 0.001), i.e. they were more rigid, less open to diversity, less tolerant, tending more to perfection and providing more socially acceptable answers. These characteristics highly correlated with experiencing infertility as an extreme stress and were also in relationship with maternal age. Conclusions. The development of ICSI children is within the normal range; however, they tend to be at increased risk for emotional problems. There exists a strong correlation between the child’s characteristics and the mother’s personality that has been marked with the experience of infertility and their age

NEW CLASSIFICATION OF SPERM MORPHOLOGY BEFORE ICSI AT MAGNIFICATION OF X6000

Background. At conventional ICSI, sperm to be injected into the oocyte is chosen at magnification of 400 times which does not permit abnormalities of fine sperm structures to be seen. By using a new method, intracytoplasmic morphologically selected injection (IMSI), motile sperm with a normal head, base and no vacuoles present in the head can be selected at magnification of 6000 times. Vacuoles in the sperm head reflect the damage of nucleus and/or DNA (fragmented or single-stranded DNA). Spermatozoa with vacuoles in the head have lower fertilization capacity and lead to abnormal embryo development, unsuccessful implantation, or spontaneous abortion. Methods. The aim of this study was to evaluate the percentage of men with abnormal sperm morphology (teratozoospermia) included in the ICSI programme, in which it is possible to find a morphologically normal sperm without vacuoles in the head using IMSI method. In each selected sperm we evaluated the morphology of head (normal = 2 points), base (normal = 1 point) and presence of vacuoles in the head (one small or no vacuoles = 3 points). Totally normal sperm (or with one small vacuole in the head) was allotted 6 points. According to this evaluation, sperm were divided into three classes: class I (6–4 points), class II (3–1 points) and class III (0 points). Sperm of class I and II were suitable for injection into the oocyte, whereas sperm of class III were not. Results. Before ICSI, the prepared semen of 13 patients which had teratozoospermia (alone or in combination with other semen abnormalities) was evaluated by the IMSI method. Sperm of class I was found in 5 men (38 %), and sperm of class II in the remaining men (62 %). Sperm of class III was found in all men. On average, 0.7 sperm of class I and 6.1 sperm of class II per man with teratozoospermia were found. Conclusions. Morphologically normal sperm without vacuoles in the head can be found in less than half of men undergoing the ICSI programme. Further research will show, whether it is possible to improve the clinical results of the ICSI method by the sperm selection with the IMSI method

Genetic variation in circadian rhythm genes CLOCK and ARNTL as risk factor for male infertility.

BACKGROUND: The circadian system has a major role in maintaining homeostasis and proper body functions including reproductive capacity. The aim of this study was to examine whether there is an association between genetic variability in the primary clock genes CLOCK and ARNTL and male infertility in humans. METHODOLOGY/PRINCIPAL FINDINGS: We performed a case-control study, where we searched for an association between polymorphisms of CLOCK and ARNTL genes and male infertility in 961 Slovenian and Serbian Caucasian men. The study group consisted of 517 patients with idiopathic infertility and a control group of 444 fertile men. A statistically significant difference was found in genotype distribution between the two groups in the CLOCK gene: rs11932595 (p = 6·10(-5), q = 4·10(-4), OR equaled 1.9 with 95% CI 1.4-2.7), rs6811520 (p = 2·10(-3), q = 8·10(-3), OR = 1.7 with 95% CI 1.2-2.2) and rs6850524 (p = 0.01, q = 0.02, OR = 1.4 with 95% CI 1.1-1.9). Further analyses of haplotypes were consistent with genotyping results. CONCLUSIONS/SIGNIFICANCE: We provide evidence that genetic variability in the CLOCK gene might be associated with male infertility warranting further confirmation and mechanistic investigations

Frequencies and distribution of probable haplotypes in the patient and control groups as predicted by haplo.stats.

<p>Frequencies and distribution of probable haplotypes in the patient and control groups as predicted by haplo.stats.</p

Genotype and allelic distribution of the CLOCK and ARNTL polymorphisms of the 517 infertile men and 444 fertile controls.

*<p>ARNTL SNP rs12363415 showed a departure from the Hardy-Weinberg equilibrium and was excluded.</p