Mature oocyte dysmorphisms may be associated with progesterone levels, mitochondrial DNA content, and vitality in luteal granulosa cells.

PURPOSE: To identify whether follicular environment parameters are associated with mature oocyte quality, embryological and clinical outcomes. METHODS: This retrospective study examined 303 mature oocytes from 51 infertile women undergoing ICSI cycles between May 2018 and June 2021. Exclusion criteria consisted of advanced maternal age (> 36 years old), premature ovarian failure, obesity in women, or use of frozen gametes. Luteal granulosa cells (LGCs) were analyzed for mitochondrial DNA/genomic (g) DNA ratio and vitality. The relationships between hormone levels in the follicular fluid and oocyte features were assessed. Quantitative morphometric measurements of mature oocytes were assessed, and the association of LGC parameters and oocyte features on live birth rate after single embryo transfer was examined. RESULTS: Results indicated an inverse correlation between the mtDNA/gDNA ratio of LGCs and the size of polar body I (PBI). A 4.0% decrease in PBI size was observed with each one-unit increase in the ratio (p = 0.04). Furthermore, a 1% increase in LGC vitality was linked to a 1.3% decrease in fragmented PBI (p = 0.03), and a 1 ng/mL increase in progesterone levels was associated with a 0.1% rise in oocytes with small inclusions (p = 0.015). Associations were drawn among LGC characteristics, perivitelline space (PVS) debris, cytoplasmic inclusions, PBI integrity, and progesterone levels. Certain dysmorphisms in mature oocytes were associated with embryo morphokinetics; however, live birth rates were not associated with follicular parameters and oocyte quality characteristics. CONCLUSION: Follicular markers may be associated with mature oocyte quality features.

Identifying predictors of Day 5 blastocyst utilization rate using an artificial neural network.

RESEARCH QUESTION: Can artificial intelligence identify predictors of an increased Day 5 blastocyst utilization rate (D5BUR), which is one of the most informative key performance indicators in an IVF laboratory? DESIGN: This retrospective, multicentre study evaluated six variables for predicting D5BUR using an artificial neural network (ANN): number of metaphase II (MII) oocytes injected (intracytoplasmic sperm injection); use of autologous/donated gametes; maternal age at oocyte retrieval; sperm concentration; progressive sperm motility rate; and fertilization rate. Cycles were divided into training and testing sets through stratified random sampling. D5BUR on Day 5 was grouped into <60% and ≥60% as per the Vienna consensus benchmark values. RESULTS: The area under the receiver operating characteristic curve (AUC) to predict the D5BUR groups was 80.2%. From the ANN model, all six independent variables were found to be of significant value for the prediction of D5BUR (P<0.0001), with the most important variable being the number of MII oocytes injected. Investigation of the effect of MII oocytes injected on D5BUR indicated an inverse correlation, with injection of an increasing number of MII oocytes resulting in a decreasing D5BUR (r=-0.344, P<0.001) and injection of up to six oocytes resulting in D5BUR ≥60%. CONCLUSION: The number of MII oocytes injected is the most important predictor of D5BUR. Exploration of additional variables and further validation of models that can predict D5BUR can guide the way towards personalized treatment and increased safety.

Do follicle-stimulating hormone receptor polymorphisms in infertile men influence intracytoplasmic sperm injection outcomes? A prospective cohort study.

The follicle-stimulating hormone (FSH) and its receptor regulate the quantity and quality of spermatozoa production. Several studies have analyzed the effect of single nucleotide polymorphisms (SNPs) in exon 10 of the FSH receptor (FSHR) on basic semen parameters without yet reaching a firm consensus. The aim of this study was to evaluate the effect of p.Thr307Ala and p.Asn680Ser polymorphisms in exon 10 of the FSHR gene, in infertile men, on intracytoplasmic sperm injection (ICSI) outcomes. This study was conducted between March 2019 and February 2020 on infertile couples undergoing ICSI at Al Hadi Laboratory and Medical Center, Lebanon. Couples with severe infertility factors that may impair gametogenesis/embryogenesis (e.g. advanced maternal age, premature ovarian failure, underwent gonadotoxic treatments, etc.) were excluded from the study. Semen and blood samples were collected from infertile men on the day of oocyte collection. Infertile men (n = 173) were screened for FSHR variants using polymerase chain reaction-restriction fragment length polymorphism. Moreover, fertilization rates, embryo quality, and pregnancy outcomes were evaluated. Higher sperm concentrations were found in the p.Thr307Ala group than the p.Thr307Thr (P < 0.01) and p.Ala307Ala (P < 0.05) groups. Furthermore, fertilization rate was significantly lower in the p.Ala307Ala genotype than in the p.Thr307Thr genotype (P < 0.05). We showed that FSHR variants in infertile men undergoing ICSI could affect sperm concentration, motility, and fertilization rates. Therefore, it will be important to confirm these results in further studies using a larger sample size.

Stress Management during the Intracytoplasmic Sperm Injection Cycle May Slow Down First Embryo Cleavage and Accelerate Embryo Compaction: A Pilot Randomized Controlled Trial.

INTRODUCTION: A firm consensus on the effectiveness of psychological interventions during infertility treatment has not been reached yet in terms of mental health and pregnancy rates. Moreover, the influence of these interventions on embryo cleavage kinetics has not been investigated. OBJECTIVE: The aim of this work was to study whether stress management in couples undergoing an intracytoplasmic sperm injection (ICSI) cycle influences stress levels, mitochondrial DNA (mtDNA) levels in granulosa cells, and cleavage-stage embryos. METHODS: Infertile couples were randomized into a treatment as usual (TAU) group (n = 30) and stress management program (SMP) group (n = 29) at the beginning of an ICSI cycle. Couples in the SMP group attended education and relaxation sessions at each visit to the clinic for folliculometry. The perceived stress scale (PSS) was used to assess stress levels at the beginning and end of the cycle. Moreover, mtDNA levels of granulosa cells and embryo morphokinetics were evaluated. RESULTS: Post-intervention, women in the SMP group had significantly lower PSS scores than their initial PSS (p < 0.001; effect size, ES = 0.5) and than the final PSS of the TAU group (p = 0.02; ES = 0.09). Additionally, mtDNA levels were significantly lower in luteal granulosa cells of the SMP group than the TAU group (p = 0.02). An earlier time of pronuclei appearance (p = 0.03) and time to 2 cells (p = 0.015) and a faster time to full compaction (p = 0.045) were detected in the embryos of the SMP group compared with the TAU group. CONCLUSION(S): The implemented program may reduce stress levels, retard first embryo cleavage, and accelerate embryo compaction. Further studies with an active control group are needed to confirm these results.

What are the effects of vitamin C on sperm functional properties during direct swim-up procedure?

SummaryDirect swim-up procedure is widely used to separate the motile competent spermatozoa from the antioxidant-rich semen. Subsequently, spermatozoa become more vulnerable to reactive oxygen species (ROS) due to their cytological characteristics. The effect of vitamin C, a highly concentrated antioxidant in the semen, on direct swim-up-enriched sperm population is not fully investigated. Therefore, the aim of the present study was to assess the effect of vitamin C on sperm functional properties during direct swim-up procedure. Semen samples were collected from 22 participants. Each semen sample was divided into several aliquots. The first portion was overlaid with sperm medium without ascorbic acid (0 µM AA). The second and third fractions were overlaid with sperm medium supplemented with 300 µM and 600 µM AA; respectively. After 1 h of incubation, basic sperm parameters, intracellular ROS levels, acrosome reaction, chromatin integrity, and glucose uptake were assessed. Swim-up without AA significantly increased the percentage of ROS(+) spermatozoa compared with the raw semen (P<0.01). Interestingly, swim-up with 300 µM AA did not increase the percentage of ROS(+) sperm compared with the raw semen. In parallel, the percentage of sperm with altered chromatin integrity was significantly lower in the 300 µM AA group compared with that in the raw semen (P<0.05). These findings suggest that supplementation of vitamin C to sperm medium could be beneficial for direct swim-up-derived spermatozoa.

Hydatid disease: the Lebanese contribution.

Hydatid disease (HD) or cystic echinococcosis (CE) has been an important zoonotic disease causing medical, economic and public health problems in many parts of the world, including South America, North Africa, Turkey, and Middle Eastern countries. Humans as well as animals, primarily sheep and cattle, are infected by the ingestion of food, usually leafy vegetables, contaminated with the eggs (oncospheres) of the dog tapeworm Echinococcus granulosus. Hydatid cysts, which are the larval stage of the parasite, are located mostly in the liver and lungs of the infected host. Because of its chronic endemicity in Lebanon and neighboring countries, this disease has constituted an integral part of research studies conducted by medical doctors and researchers in Lebanon, mostly spearheaded by those at the American University of Beirut (AUB) and its medical center (AUBMC) since the early turn of the last century (1920s). Over 130 wide ranging studies were published; some were innovative e.g. the introduction of the once famous Indirect Haemagglutination (IHA) test for serodiagnosis, and the use of dilute cetrimide as a protoscolicidal agent during surgery. Although the incidence of HD is decreasing in our country, it has acquired increasing public health concern and is considered as an emerging or re-emerging disease in many parts of the world. In this review, we shed light on the numerous studies/publications done in Lebanon as a tribute to those researchers who have impacted the literature of HD in many aspects. The latter include epidemiology and ways of transmission, clinical features and radiological tools for diagnosis, serodiagnosis and immunology, and investigation of different therapeutic modalities for different aspects of the disease. Moreover, consolidating these studies in this review would hopefully represent the historic foundation for interested researchers and investigators, especially in this country, to pursue and build on such studies. The advances in technology, and the availability and utilization of new methodologies will hopefully help find more reliable and efficient ways for the diagnosis, and management of this disease.

Prevalence and Time Trends of Low Serum B12 Levels and Inadequate B12 Dietary Intake in Lebanese Adults amidst the Food Insecurity Situation: Findings from a Nationally Representative Cross-Sectional Study.

Rising food insecurity (FI) and the increased prices of animal-based foods could compromise vitamin B12 (B12) intake and serum levels in food-insecure people. Our study aims to determine the prevalence of low levels of serum B12 and its inadequate dietary intake among a nationally representative sample of Lebanese adults aged 18 to 64 years, while exploring the impact of FI and identifying other potential predictors. The B12 intake was assessed using a food frequency questionnaire and the mean of two non-consecutive 24 h recalls. The B12 serum levels were also examined. To examine the time trends in the B12 serum levels and dietary intake, the data from a large representative digitized database of 6290 participants were collected, along with the examination of FAOSTAT food consumption data both before and during the economic crisis period. Our findings revealed that 47.3% of households in Lebanon experienced FI. The inadequate intake of B12 food sources was prevalent in 52.5% of participants, and 61.1% presented with low (23%) or intermediate (38.1%) levels of serum B12. The food-secure households had a higher proportion of B12 intake from meats (p = 0.004), while traditional foods contributed more to food-insecure households (p = 0.000). The participants who were female, unemployed, food-insecure, and had low dietary diversity were about two to three times more likely to have inadequate B12 intake. The predictors of low levels of serum B12 included residing in Beqaa (OR = 2.856 and 95% CI = 1.018-8.01) and having inadequate B12 intake (OR = 1.788 and 95% CI = 1.2-2.65). The findings from observing the time trends in the consumption and the serum levels of B12 indicate a considerable decline in the consumption of most animal-based foods compared to the pre-crisis period, associated with a decline in the serum levels of B12. In conclusion, an alarming prevalence of low dietary intake and low serum levels of B12 was revealed among Lebanese adults, and the indirect effect of FI on B12 serum levels mediated through B12 intake was inferred.

Functional histology of human scrotal wall layers and their overlooked relation with infertility: a narrative review.

Male infertility currently contributes to nearly half of the reported infertility cases. Scrotal wall layers play a cardinal role in regulating testicular physiology. However, few studies have focused on the functional histology of these layers and their relations with infertility in humans. The objective of the present narrative review is to collate novel insights into the functional histology of the human scrotal wall layers and their relation with infertility. The data was extracted from articles published between 1946 and 2021. The study was performed between January and December 2021. 71 original studies have been included in this review. Despite the fact that few studies have presented detailed functional histology of the human scrotal wall layers, this narrative review elucidates the possible influence of scrotal histology on infertility. Scrotal wall layers-associated pathologies may induce infertility by various mechanisms. They can impose mechanical forces that may affect the testicular histology and stimulate testicular inflammation. Moreover, they may induce testicular hyperthermia. Various unanswered clinical questions have been identified in this narrative review. More clinical studies are needed to assess the effect of alterations in the components of the scrotal wall layers on fertility (e.g., due to the exposure to metabolic and/or psychological stressors). In addition, testing the effectiveness of various pharmacological/surgical interventions to treat scrotal wall layers-associated pathologies will provide more insights into infertility treatment.

Lactobacillus plantarum secretions may exert a cryoprotective effect on human sperm motility: A prospective in vitro study.

BACKGROUND: Semen cryopreservation is a widely used procedure for fertility preservation, despite some level of cryodamage that may occur in spermatozoa after thawing. However, there is some evidence that lactobacilli, one of the bacteria found in semen, might benefit sperm quality. OBJECTIVES: This study aims to determine whether the addition of Lactobacillus plantarum secretions to sperm freezing medium has an impact on sperm motility, morphology, and DNA fragmentation. MATERIALS AND METHODS: This is a prospective auto-controlled study. It was conducted on 30 raw semen samples from 30 infertile men attending a fertility center for semen analysis. Before freezing, all the samples were analyzed for motility, morphology, and DNA fragmentation percentages. Each sample was then divided equally into three aliquots. Cryopreservation was performed on each aliquot using one of the following three media: without Lactobacillus plantarum secretions (control group) or with 107 or 108 colony-forming units/mL Lactobacillus plantarum secretions. Sperm motility, morphology, and DNA integrity were evaluated after the cryopreservation media were added and after semen thawing. RESULTS: The results of this study indicated that after thawing, no statistically significant decrease in progressive motility and non-progressive percentages were detected in the sperm freezing medium supplemented with 108 colony-forming units/mL Lactobacillus plantarum secretions than the fresh raw semen. Moreover, multivariate linear regression model analyses showed that the progressive motility (p = 0.02), non-progressive motility (p = 0.016), and non-motile spermatozoa (p = 0.012) percentages were significantly decreased in the freezing medium (without Lactobacillus plantarum secretions) compared to the fresh raw semen. DISCUSSION AND CONCLUSION: To the best of our knowledge, this is the first study showing that Lactobacillus plantarum secretions had a cryoprotective effect on sperm motility when added to the sperm freezing medium. Furthermore, Lactobacillus plantarum secretions were found to protect sperm DNA integrity more effectively than the freezing medium without Lactobacillus plantarum secretions in non-normozoospermia group. Cryopreservation procedures must therefore be optimized to minimize any iatrogenically induced sperm DNA damage, given the correlation between sperm DNA damage and increased mutation loads in progeny.

Lebanese SARS-CoV-2 genomics: 24 months of the pandemic.

The COVID-19 pandemic continues to pose a global health concern, despite the ongoing vaccination campaigns, due to the emergence and rapid spread of new variants of the causative agent SARS-CoV-2. These variants are identified and tracked via the marker mutations they carry, and the classification system put in place following tremendous sequencing efforts. In this study, the genomes of 1,230 Lebanese SARS-CoV-2 strains collected throughout 2 years of the outbreak in Lebanon were analyzed, 115 of which sequenced within this project. Strains were classified into seven GISAID clades, the major one being GRY, and 36 Pango lineages, with three variants of concern identified: alpha, delta and omicron. A time course distribution of GISAID clades allowed the visualization of change throughout the two years of the Lebanese outbreak, in conjunction with major events and measures in the country. Subsequent phylogenetic analysis showed the clustering of strains belonging to the same clades. In addition, a mutational survey showed the presence of mutations in the structural, non-structural and accessory proteins. Twenty five (25) mutations were labeled as major, i.e. present in more than 30% of the strains, such as the common Spike_D614G and NSP3_T183I. Whereas 635 were labeled as uncommon, i.e. found in very few of the analyzed strains as well as GISAID records, such as NSP2_I349V. Distribution of these mutations differed between 2020, and the first and the second half of 2021. In summary, this study highlights key genomic aspects of the Lebanese SARS-CoV-2 strains collected in 2020, the first year of the outbreak in Lebanon, versus those collected in 2021, the second year of COVID-19 in Lebanon.

Knowledge, Attitudes, and Practices Related to Dietary Supplementation, before and during the COVID-19 Pandemic: Findings from a Cross-Sectional Survey in the Lebanese Population.

At the start of 2020, a new coronavirus (COVID-19) invaded the world leading to the death of 3.92 million people. Sadly, to date, no remedy has been discovered for this virus. Preventive vaccines have been under investigation, but were unavailable until December 2020. Clinical deficiencies of nutrients may increase susceptibility to infections. This knowledge may have provided an incentive for some dietary supplement (DS) manufacturers to advertise their products as COVID-19 preventatives or cures without any substantiation, targeting mainly social media fans. The objective of this research was to assess the usage, knowledge and attitudes towards dietary supplementation before and during the COVID-19 pandemic among Lebanese people. A cross-sectional study was conducted based on a convenience sample (N = 2966) and information from participants aged 18 years and above was collected about periods before and during the pandemic. Our findings showed that attitudes towards DSs changed when the pandemic emerged and people believed that DSs can improve their health and strengthen their immunity. Despite the rise in DS prescription by healthcare professionals, the prevalence of DS use decreased from 73.3% before the pandemic to 69.9% during the pandemic (p < 0.001). Study results declared that the weekly or the daily estimated intake had increased during the pandemic as compared to before the pandemic, from 14% to 15.6% for antioxidants (p = 0.014), from 35.3% to 42.1% for vitamin C (p < 0.001), from 35.5% to 41% for vitamin D (p < 0.001), from 15.2% to 17.5% for vitamin E (p = 0.002), and from 18.8% to 29.3% for zinc (p < 0.001) and other vitamins and minerals (from 9% to 10.9%, p < 0.001). Binary logistic regression indicated that the use of DSs during the pandemic was 4 times higher among those infected with COVID-19, 30 times higher among those who used to take DSs before pandemic, and 1.5 times higher among those who worked in the medical sector. To conclude, there is a crucial need to increase awareness among Lebanese people regarding the use of DSs.

Differential impact of four sperm preparation techniques on sperm motility, morphology, DNA fragmentation, acrosome status, oxidative stress, and mitochondrial activity: A prospective study.

BACKGROUND: Suboptimal human semen handling in vitro may induce sperm damage. However, the effects of semen swim-up, pellet swim-up, density gradient, and density gradient followed by SU on sperm motility, morphology, DNA fragmentation, acrosome reaction, intracellular reactive oxygen species, and mitochondrial activity were not fully understood. OBJECTIVES: To study the impact of four sperm preparation techniques on sperm functional parameters. MATERIALS AND METHODS: This study was conducted on 60 infertile men with a minimum sperm concentration of 20 × 106 /ml and total sperm motility of ≥30%. Each raw semen sample was divided into four aliquots. Each aliquot was prepared by one of the tested techniques. Various sperm characteristics were assessed before and after sperm preparation. RESULTS: Density gradient and density gradient followed by SU resulted in significantly higher DNA fragmentation percentages compared with semen swim-up (p < 0.001 and p < 0.001, respectively) and pellet swim-up (p < 0.001 and p < 0.001, respectively). Significantly higher percentages of spermatozoa with intact acrosome were detected in semen swim-up (p < 0.001) and pellet swim-up (p < 0.001) compared with raw semen. The percentage of reactive oxygen species-positive spermatozoa was significantly higher after pellet swim-up (p < 0.001), density gradient (p < 0.001), and density gradient followed by SU (p < 0.001) than raw semen. In addition, the percentages of 100% stained midpiece (active mitochondria) were significantly higher in semen swim-up (p < 0.001) and pellet swim-up (p < 0.001) compared with raw semen. DISCUSSION AND CONCLUSION: To the best of our knowledge, this is the first report comparing the impact of these techniques on various sperm functional parameters. Semen swim-up was more effective than density gradient in selecting better spermatozoa in terms of DNA integrity, reactive oxygen species levels, acrosome status, and mitochondrial activity. Randomized clinical trials comparing these four techniques are required to test their impact on embryo development and pregnancy outcomes.

SARS-CoV-2 Variants in Lebanon: Evolution and Current Situation.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has seen a worldwide spread since its emergence in 2019, including to Lebanon, where 534,968 confirmed cases (8% of the population) and 7569 deaths have been reported as of 14 May 2021. With the genome sequencing of strains from various countries, several classification systems were established via genome comparison. For instance, the GISAID clades classification highlights key mutations in the encoded proteins that could potentially affect the virus' infectivity and transmission rates. In this study, 58 genomes of Lebanese SARS-CoV-2 strains were analyzed, 28 of which were sequenced for this study, and 30 retrieved from the GISAID and GenBank databases. We aimed to classify these strains, establish their phylogenetic relationships, and extract the mutations causing amino acid substitutions within, particularly, the structural proteins. The sequenced Lebanese SARS-COV-2 strains were classified into four GISAID clades and 11 Pango lineages. Moreover, 21 uncommon mutations in the structural proteins were found in the newly sequenced strains, underlining interesting combinations of mutations in the spike proteins. Hence, this study constitutes an observation and description of the current SARS-CoV-2 genetic and clade situation in Lebanon according to the available sequenced strains.

Optimization of The Cell Aggregates Method for Isolation and Purification of Human Granulosa Cells from Follicular Fluid.

BACKGROUND: Aspirated ovarian follicular fluids (FF) contain luteal granulosa cells (LGCs) and other contaminating cell types. Several strategies, such as the antibody binding methods, the flask method, the cell strainer and positive selection of granulosa aggregates after density gradient (DG) centrifugation, were tested as LGC purification methods. Each of these strategies has its own advantages and disadvantages. Positive selection of granulosa aggregates after DG centrifugation is simple, rapid and efficient in terms of LGC recovery. However, it results in a low purity. Here, we aimed to test whether modifying the traditional protocol by collecting the aggregates from the FF, before the DG centrifugation could decrease the percentage of contaminating cells. MATERIALS AND METHODS: In the present prospective study, 32 FF, from 32 women,were randomly assigned into one of the two purification techniques: positive selection of granulosa aggregates from the FF, after DG centrifugation (DG/ Agg, n=16) or positive selection of granulosa aggregates from the FF, before DG centrifugation (Agg/DG, n=16). At the end of each procedure cell count, vitality, morphology and purity of the cell suspension were evaluated. RESULTS: No significant difference was detected in the total number of GCs between DG/Agg and Agg/DG (P>0.05). However, higher percentage of GCs with normal morphology was detected in Agg/DG compared to DG/Agg (P<0.001). Moreover, lower percentages of white blood cells (P<0.01), red blood cells (P<0.001) and epithelial cells (P<0.01) were identified in Agg/DG compared to DG/Agg. CONCLUSION: Here we showed that positive selection of granulosa aggregates from the FF prior to DG technique had a higher purity compared to the traditional protocol. Thus, it could be a method of choice to prepare GCs for research purposes in clinical in vitro fertilization settings.

Infantile Cytomegalovirus-Associated Severe Warm Autoimmune Hemolytic Anemia: A Case Report.

Autoimmune hemolytic anemia is a rare hematologic entity in children. Etiologies are mainly viruses or bacteria. We describe here a case of severe warm autoimmune hemolytic anemia (IgG- and C3d-positive direct antiglobulin test) in an immunocompetent 6-month-old infant with acute Cytomegalovirus infection that responded well to corticotherapy and intraveneous immunoglobulins without using blood component transfusion. This case demonstrates the importance of recognizing CMV in infantile Autoimmune Hemolytic Anemia, especially because hemolysis can be severe and lethal.