Chromosomal abnormalities and Y chromosome microdeletions in infertile men from Morocco.

BACKGROUND: Male infertility is responsible for 50% of infertile couples. Thirty percent of male infertility is due to cytogenetic and genetic abnormalities. In Arab and North African populations, several studies have shown the association of these chromosomal abnormalities with male infertility. Our objective is to evaluate the frequency of chromosomal abnormalities and Y chromosome microdeletions in infertile men from Morocco. METHODS: A total of 573 Moroccan infertile men (444 azoospermic and 129 oligozoospermic men) referred for cytogenetic analysis to the Department of Cytogenetics of the Pasteur Institute of Morocco, were screened for the presence of chromosomal abnormalities and Y chromosome microdeletions. RESULTS: Chromosomal abnormalities accounted for approximately 10.5% (60/573). Fifty six cases among them have sex chromosome abnormalities (93.34%), including Klinefelter's syndrome in 41 patients (68.34%). Autosomal chromosome abnormalities (6.66%) were observed in 4 patients. Chromosomal abnormalities were more prevalent in azoospermic men (13.06%) than in oligospermic men (1.55%). Y microdeletions were detected in 16 of 85 patients (AZFc: 14.12%, AZFbc: 4.70%), most of them where azoospermic men with no chromosomal abnormality. CONCLUSIONS: These results highlighted the need for efficient molecular genetic testing in male infertility diagnosis. In addition, a genetic screening should be performed in infertile men before starting assisted reproductive treatments.

Common loss-of-function variants of the epidermal barrier protein filaggrin are a major predisposing factor for atopic dermatitis.

Atopic disease, including atopic dermatitis (eczema), allergy and asthma, has increased in frequency in recent decades and now affects approximately 20% of the population in the developed world. Twin and family studies have shown that predisposition to atopic disease is highly heritable. Although most genetic studies have focused on immunological mechanisms, a primary epithelial barrier defect has been anticipated. Filaggrin is a key protein that facilitates terminal differentiation of the epidermis and formation of the skin barrier. Here we show that two independent loss-of-function genetic variants (R510X and 2282del4) in the gene encoding filaggrin (FLG) are very strong predisposing factors for atopic dermatitis. These variants are carried by approximately 9% of people of European origin. These variants also show highly significant association with asthma occurring in the context of atopic dermatitis. This work establishes a key role for impaired skin barrier function in the development of atopic disease.

Assessment of Knowledge about First Aid Methods, Diagnosis, and treatment of scorpion stings among health workers in Ouarzazate region, Morocco: A Cross-Sectional Study.

Scorpion envenomation (SE) is a critical and immediate medical emergency. It represents a major public health concern with significant cause of illness and death, particularly in rural regions where scorpions are more prevalent. Of note, administering appropriate first aid to scorpion sting victims is one of the most effective ways to reduce mortality. Therefore, it is crucial to understand the level of knowledge of the healthcare workers in managing such cases. This work aims to assess the levels of knowledge on the diagnosis and management of scorpion stings, as well as the attitudes toward scorpion stings, among doctors and nurses in the Ouarzazate region of Morocco. Our results showed that the prevalent health care age group is 30-39 years, and in terms of experience, the majority had less than 5 years. Additionally, the majority of the participants had no knowledge regarding scorpions' identification. Moreover, only half participants (healthcare workers) received basic training in the management of scorpion stings and almost all of them have not benefited from continuing training. Hence, the current study showed that the understanding of scorpion sting management among healthcare providers in Ouarzazate province is inadequate to ensure proper care for sting patients, and thus including scorpion sting management in the university programs of health care provides along with clinical practice are highly required.

Multifactorial hypovitaminosis D among women in the South-East Oases of Morocco.

This study assessed the vitamin D status among women in South-East Oases of Morocco. Two hundred thirty-three healthy volunteer women were recruited at five urban and rural health centers. Hypovitaminosis D is very common in this region and correlated to age, monthly income, skin color, daily sun exposure, BMI, and body fat percentage. PURPOSE: This study assessed the vitamin D status among women in the Draa-Tafilalet region and its relationship with clinical, anthropometric, and behavioral parameters. METHODS: The study was carried out with 233 women in consultation at five urban and rural health centers in the South-East region of Morocco. The data collected relate to age, monthly income, educational level, BMI, body fat percentage, daily sun exposure, physical activity level, veil wear, and skin color. The plasma determination of vitamin D was measured by immunofluorescence. Statistical analyses were performed using Python and Jamovi. RESULTS: The median 25(OH)D plasma concentration was 9.95 ng/mL (IQR: 8-13.18). A total of 50.64% of women had 25(OH)D levels below 10 ng/ml and 47.21% had levels between 10 and 30 ng/mL. Statistical tests showed an association between hydroxyvitamin D plasma levels and age (r = - 0.139; p = 0.034), monthly income (p = 0.001), BMI (r = - 0.200; p = 0.002), body fat percentage (r = - 0.131; p = 0.049), daily sun exposure (r = 0.165; p = 0.012), and skin color (p < 0.001). Binomial logistic regression showed that darker skin was associated with vitamin D deficiency (< 10 ng/mL). It also showed that high income and longer sun exposure could be protecting factors against vitamin D deficiency. CONCLUSION: The plasma concentrations of vitamin D in women in the Draa-Tafilalet region were low compared to expected results for a high sunshine time area.

Epidemiological Profile of Drug Overdose Reported in South-East Morocco from 2004 to 2016.

BACKGROUND: Drug overdose, either accidental or intentional, is the most common cause of acute poisoning worldwide. OBJECTIVE: The aim of this study was to determine the epidemiological profile of drug poisoning recorded in the south east of Morocco and to identify the proportion of intentional versus accidental drug overdose. METHODS: This was an epidemiological study of 180 cases of medicinal poisoning registered with the Provincial Delegation of Health in Errachidia between January 2004 and December 2016. Information on demographic and drug overdose characteristics was obtained from the regional poison center. Drugs were categorized according to the anatomical therapeutic chemical (ATC) classification system. RESULTS: Adults were the most affected group, with a median age of 21 years and a sex ratio of three females to every male. Drug poisoning mainly occurred in urban areas (83% of cases). Regarding clinical signs, 55.2% of patients presented with digestive signs and 27.6% with neurological signs. Other signs were also present: respiratory (5.1%), combined neurological and digestive (4.5%), cardiovascular (3.8%), and general (3.8%). Women represented 88.9% of those who had intentionally overdosed and 64.3% of those who had unintentionally overdosed. Benzodiazepine derivatives and other related drugs were involved in 21.5% of cases of drug poisoning, with other drugs found in patients with drug overdose as follows: paracetamol 3.3%, ethinyl estradiol/levonorgestrel 5%, and cyproheptadine 1.6%. CONCLUSION: Our results indicate that the number of reported cases of drug poisoning in south-east Morocco increased between 2004 and 2016. The intentional use of drugs in overdose was mostly  among adults, especially women. The drugs involved were predominantly psycholeptic drugs, followed by analgesics. Mortality was low, but investigation in a representative sample will show  the real severity and outcomes of drug overdoses.

No association between T222P/LGR8 mutation and cryptorchidism in the Moroccan population.

BACKGROUND: Cryptorchidism is the most common genital anomaly in men. The INSL3/LGR8 system is involved in testicular descent via gubernacular development. INSL3 binds with high affinity to its receptor LGR8 and receptor activation is associated with cAMP signaling. Analysis of human INSL3 and LGR8 mutations confirms that some cases of cryptorchidism are caused by mutations in these genes. The T222P mutation is the only one within the LGR8 gene associated with the cryptorchidism phenotype. A strong association of the T222P mutation with cryptorchidism was found in an Italian population. Due to the same mutation being found in patients within the Mediterranean area, a possible founder effect of this mutation is supposed. METHODS: We screened 109 patients with cryptorchidism and 250 controls in a Moroccan population. RESULTS: We found that 3 of the 109 patients tested carry the T222P mutation and 4 individuals in the control group also carry the mutation. CONCLUSIONS: Our results show in fact that the same mutation is present in the Moroccan population, but an association between cryptorchidism and the T222P mutation was not found.

AZF microdeletions and partial deletions of AZFc region on the Y chromosome in Moroccan men.

AIM: To evaluate for the first time the frequency of Y chromosome microdeletions and the occurrence of the partial deletions of AZFc region in Moroccan men, and to discuss the clinical significance of AZF deletions. METHODS: We screened Y chromosome microdeletions and partial deletions of the AZFc region of a consecutive group of infertile men (n = 149) and controls (100 fertile men, 76 normospermic men). AZFa, AZFb, AZFc and partial deletions of the AZFc region were analyzed by polymerase chain reaction (PCR) according to established protocols. RESULTS: Among the 127 infertile men screened for microdeletion, four subjects were found to have microdeletions: two AZFc deletions and two AZFb+AZFc deletions. All the deletions were found only in azoospermic subjects (4/48, 8.33%). The overall AZFc deletion frequency was low (4/127, 3.15%). AZF microdeletions were not observed in either oligoasthenoteratozoospermia (OATS) or the control. Partial deletions of AZFc (gr/gr) were observed in a total of 7 of the 149 infertile men (4.70%) and 7 partial AZFc deletions (gr/gr) were found in the control group (7/176, 3.98%). In addition, two b2/b3 deletions were identified in two azoospermic subjects (2/149, 1.34%) but not in the control group. CONCLUSION: Our results suggest that the frequency of Y chromosome AZF microdeletions is elevated in individuals with severe spermatogenic failure and that gr/gr deletions are not associated with spermatogenic failure.

Mutation rate at 17 Y-STR loci in "Father/Son" pairs from moroccan population.

Precise knowledge of mutation rate at Y-STRs loci is essential for a correct evaluation of typing results in forensic casework and specially kinship genetic studies. In this study, we have examined 252 confirmed and unrelated father/son sample pairs from Moroccan population using the 17 Y-STR markers DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385a, DYS385b, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, and Y-GATA-H4 of the AmpFlSTR Yfiler™ kit used in routine casework. We observed a total of 15 single repeat mutations between fathers and sons as mutational events. Nine mutations resulted in the gain of a repeat in the son and six resulted in a loss of a repeat. The average mutation rate in the studied sample is 3.5×10(-3) (95% CI 2-5.8×10(-3)). Furthermore, Y-STRs mutation occurrence seems to be 4 times more frequent than autosomal STRs mutation in this sample.

Association of spermatogenic failure with the b2/b3 partial AZFc deletion.

Infertility affects around 1 in 10 men and in most cases the cause is unknown. The Y chromosome plays an important role in spermatogenesis and specific deletions of this chromosome, the AZF deletions, are associated with spermatogenic failure. Recently partial AZF deletions have been described but their association with spermatogenic failure is unclear. Here we screened a total of 339 men with idiopathic spermatogenic failure, and 256 normozoospermic ancestry-matched men for chromosome microdeletions including AZFa, AZFb, AZFc, and the AZFc partial deletions (gr/gr, b1/b3 and b2/b3).AZFa and AZFc deletions were identified in men with severe spermatogenic failure at similar frequencies to those reported elsewhere. Gr/gr deletions were identified in case and control populations at 5.83% and 6.25% respectively suggesting that these deletions are not associated with spermatogenic failure. However, b2/b3 deletions were detected only in men with spermatogenic failure and not in the normospermic individuals. Combined with our previous data this shows an association of the b2/b3 deletion (p = 0.0318) with spermatogenic failure in some populations. We recommend screening for this deletion in men with unexplained spermatogenic failure.

Association of the MTHFR A1298C variant with unexplained severe male infertility.

The methylenetetrahydrofolate reductase (MTHFR) gene is one of the main regulatory enzymes involved in folate metabolism, DNA synthesis and remethylation reactions. The influence of MTHFR variants on male infertility is not completely understood. The objective of this study was to analyze the distribution of the MTHFR C677T and A1298C variants using PCR-Restriction Fragment Length Polymorphism (RFLP) in a case group consisting of 344 men with unexplained reduced sperm counts compared to 617 ancestry-matched fertile or normozoospermic controls. The Chi square test was used to analyze the genotype distributions of MTHFR polymorphisms. Our data indicated a lack of association of the C677T variant with infertility. However, the homozygous (C/C) A1298C polymorphism of the MTHFR gene was present at a statistically high significance in severe oligozoospermia group compared with controls (OR = 3.372, 95% confidence interval CI = 1.27-8.238; p = 0.01431). The genotype distribution of the A1298C variants showed significant deviation from the expected Hardy-Weinberg equilibrium, suggesting that purifying selection may be acting on the 1298CC genotype. Further studies are necessary to determine the influence of the environment, especially the consumption of diet folate on sperm counts of men with different MTHFR variants.

Allele frequencies and population data for 17 Y-STR loci (The AmpFlSTR® Y-filer™) in Casablanca resident population.

Allele frequencies and population data for 17 Y-STR loci included in the AmpFlSTR® Y-filer™ PCR amplification kit (Applied Biosystems, Foster City, USA), that permit the simultaneous amplification of all the markers included in the actually used European "extended haplotype", DYS19, DYS189I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385I/II, DYS438, DYS439 and also DYS437, DYS448, DYS456, DYS458, DYS635 and Y GATA H4, were obtained from a sample of 166 healthy unrelated males resident in Casablanca (from Morocco). A total of 166 haplotypes were identified, of which 142 were unique. The overall haplotype diversity for the 17 Y-STR loci reached 0.9974, and a discrimination capacity was 0.855. We report some non-standard situations, including duplications and microvariant alleles.

Y-chromosome AZFc structural architecture and relationship to male fertility.

OBJECTIVE: To determine if there is a relationship between various forms of partial AZFc deletions and spermatogenic failure. DESIGN: Case-control study. SETTING: Infertility clinic (Tenon Hospital, Paris). PATIENT(S): 557 men, comprising 364 infertile men from mixed ethnic backgrounds, and 193 men with known fertility (n = 84) and/or normospermic (n = 109). INTERVENTION(S): Characterization of 32 partial AZFc deletions. MAIN OUTCOME MEASURE(S): DAZ gene cluster divided into two families (DAZ1/2 and DAZ3/4), CDY1 gene, and Y-chromosome haplogroups. RESULT(S): We observed 18 partial AZFc deletions in 364 (4.95%) infertile men compared with 14 out of 193 (7.25%) in the control normospermic/fertile group. CONCLUSION(S): The analysis of informative Y-chromosome single nucleotide variants combined with Y-chromosome haplogroup definition enabled us to infer seven deletion classes that occur on a minimum of six Y-chromosome parental architectures. We found no relationship between either the presence or the absence of DAZ1/2, DAZ3/4, CDY1a, or CDY1b with spermatogenic failure at least on one Y-chromosome lineage. The DAZ dosage and Southern blot analyses indicated that the majority of individuals tested carried two copies of the DAZ gene, indicating a partial AZFc deletion. Our data are consistent with the hypothesis that, at least in our study populations, partial AZFc deletions may have a limited impact on fertility.

Novel mutations involving the INSL3 gene associated with cryptorchidism.

PURPOSE: Cryptorchidism affects 1% to 9% of full-term male neonates. Hypospadias is the second most frequent congenital anomaly seen in newborn males. These pathological conditions are part of the testicular dysgenesis syndrome. Insulin-like factor 3 and LGR8 (leucine-rich repeat-containing G protein-coupled receptor 8), acting as a hormone and a receptor, respectively, are involved in control of the first phase of testicular descent via gubernacular development. MATERIALS AND METHODS: The study group consisted of 184 patients, of whom 52 presented with unilateral cryptorchidism, 37 presented with bilateral cryptorchidism, 19 presented with cryptorchidism and hypospadias, 1 presented with bilateral cryptorchidism and micropenis, and 75 presented with isolated hypospadias. A control panel consisted of 270 controls, including 127 fertile, and 143 fertile noncryptorchid males. Insulin-like factor 3 mutations were analyzed by direct sequencing and restriction enzyme digestion. We analyzed the ability of the mutant insulin-like factor 3 peptides identified in this study to activate LGR8 receptor in an ex vivo assays. RESULTS: We identified 3 novel insulin-like factor 3 variants, including C-19G, V18M and R105H, in 3 of the 109 patients (2.75%) but in none of the 270 controls. The V18M mutation in the insulin-like factor 3 signal peptide had a significant deleterious effect in activating LGR8 receptor in ex vivo studies (p<0.05). To our knowledge we report the first variant in the promoter region of the insulin-like factor 3 gene in a patient with cryptorchidism in association with micropenis. CONCLUSIONS: Mutations involving the insulin-like factor 3 gene may contribute to other anomalies of male genital development, such as micropenis.

GR/GR deletions within the azoospermia factor c region on the Y chromosome might not be associated with spermatogenic failure.

No statistically significant difference in deletion frequency was found between infertile patients and a control group. It is suggested that in some populations the gr/gr deletion might be an inconsequential polymorphism.