User:Dr. Georges Raad (PhD)/sandbox

'Introduction'
Dr. Georges Raad is an experienced Lebanese Embryologist with a demonstrated history of working in the hospital & health care industry. Dr. Georges Raad is a Lecturer. Skilled in in-vitro fertilization (IVF), intracytoplamsic sperm injection (ICSI), andrology, Clinical Research, Real-Time Polymerase Chain Reaction (qPCR), next-generation sequencing, and histology. Strong healthcare services professional with a Doctor of Philosophy (PhD) focused in Reproductive Epigenetics from Université Nice Sophia Antipolis.

== 'Publications' ==

1- High level of DNA fragmentation in sperm of Lebanese infertile men using Sperm Chromatin Dispersion test
Abstract: Assessment of male fertility has been based on routine semen analysis established by the World Health Organization (WHO), evaluating sperm concentration, motility and morphology. Actually, these parameters become less reliable markers to evaluate male fertility potential. Therefore, a search for better markers has led to an increased focus on sperm chromatin integrity testing in fertility work-up and assisted reproductive technologies (ART). In this study, we evaluate sperm DNA fragmentation in 185 Lebanese infertile patients attending fertility clinics all over the country, in comparison with 30 control men of proven fertility, using the sperm chromatin dispersion test (SCD).

2- Paternal obesity: How bad is it for sperm quality and progeny health?
Abstract: There is substantial evidence that paternal obesity is associated not only with an increased incidence of infertility, but also with an increased risk of metabolic disturbance in adult offspring. Apparently, several mechanisms may contribute to the sperm quality alterations associated with paternal obesity, such as physiological/hormonal alterations, oxidative stress, and epigenetic alterations. Along these lines, modifications of hormonal profiles namely reduced androgen levels and elevated estrogen levels, were found associated with lower sperm concentration and seminal volume. Additionally, oxidative stress in testis may induce an increase of the percentage of sperm with DNA fragmentation. The latter, relate to other peculiarities such as alteration of the embryonic development, increased risk of miscarriage, and development of chronic morbidity in the offspring, including childhood cancers. Undoubtedly, epigenetic alterations (ie, DNA methylation, chromatin modifications, and small RNA deregulation) of sperm related to paternal obesity and their consequences on the progeny are poorly understood determinants of paternal obesity-induced transmission. In this review, we summarize and discuss the data available in the literature regarding the biological, physiological, and molecular consequences of paternal obesity on male fertility potential and ultimately progeny health.

3- Cryopreservation media differentially affect sperm motility, morphology and DNA integrity
Abstract: Introduction Human sperm freezing is very widely used for male fertility preservation. This procedure consists in adding cryoprotectants to the spermatozoa followed by cooling and storing the spermatozoa at a subzero temperature. Many standardized cryopreservation media are available on the market. However, these media differ in their chemical composition and there are no sufficient data to optimize their classification. Therefore, the aim of this study was to compare five commercially available sperm cryopreservation media, which have not been compared together, in terms of motility, morphology and DNA integrity. Materials and methods One‐hundred semen samples were obtained from 10 fertile participants and 90 infertile men. Each sample was evaluated before freezing for motility, morphology and DNA fragmentation index (DFI). Then, it was equally divided into five aliquots. Each aliquot was cryopreserved using one of the five media (A, B, C, D, and E). The same parameters were re‐evaluated after the addition of the cryopreservation media in the fertile group, and after sperm thawing in fertile and infertile groups. Results The results showed that the five selected cryopreservation media had negative effects on sperm motility and morphology per se. In the infertile group, the cryosurvival factor was significantly lower in cryomedium A when compared to the four other media (p < 0.001). In addition, a significantly higher percentage of sperm with coiled tail was detected in cryomedium E compared to cryomedium A (p < 0.05) and to cryomedium B (p < 0.001) after thawing, in the infertile group. Furthermore, the sperm DFI was significantly higher in cryomedia A (p < 0.001), B (p < 0.001), C (p < 0.01), D (p < 0.01) and E (p < 0.05) compared to that of the fresh semen derived from infertile participants. Conclusion This study indicates that the recovery rate of competent spermatozoa, after cryopreservation, is still critical in infertile men. Therefore, frozen semen sample should be used only when necessary.

4- Adverse effects of paternal obesity on the motile spermatozoa quality
Growing evidence suggests that paternal obesity may decrease male fertility potential. During infertility treatment with intra-cytoplasmic sperm injection (ICSI), a morphologically normal motile spermatozoon is injected into a mature egg, when possible. However, sperm motility and morphology per se do not reflect the sperm molecular composition. In this study, we aimed to assess the quality of motile spermatozoa in the context of obesity by analysing their conventional and molecular characteristics as well as their ability to promote early embryonic development. A prospective study was conducted on 128 infertile men divided into three groups: 40 lean, 42 overweight, and 46 obese men. Conventional sperm parameters (concentration, motility and morphology) and sperm molecular status (chromatin composition and integrity, 5-methycytosine (5-mC) and 5-hydroxycytosine (5-hmC) contents and oxidative stress level) were analysed on raw semen and/or on motile spermatozoa selected by density gradient or swim-up techniques. Morphokinetic analysis of the embryos derived from ICSI was performed using the Embryoviewer software. Our results showed that the motile sperm-enriched fraction from obese men exhibited higher levels of retained histones (p<0.001), elevated percentage of altered chromatin integrity (p<0.001), and decreased contents of 5-hmC (p<0.001), and 5-mC (p<0.05) levels as compared to that from lean men. Importantly, there were no statistically significant correlations between these molecular parameters and the percentages of morphologically normal motile spermatozoa. Regarding embryo morphokinetics, the CC1 (p<0.05) and CC3 (p<0.05) embryonic cell cycles were significantly delayed in the cleavage embryos of the obese group as compared to the embryos of the lean group. Our data is of particular interest because, besides demonstrating the negative impacts of obesity on motile spermatozoa molecular composition, it also highlights the possible risk of disturbing early embryonic cell cycles kinetics in the context of paternal obesity.

5- What are the effects of vitamin C on sperm functional properties during direct swim-up procedure?
Abstract: Direct 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.