Association of Renin-Angiotensin Pathway Gene Polymorphisms with COVID-19 Susceptibility and Severity in Moroccans: A Case-Control Study.

Infection by the recent SARS-CoV-2 virus causes the COVID-19 disease with variable clinical manifestations ranging from asymptomatic or mild respiratory symptoms to severe respiratory distress and multiorgan failure. The renin-angiotensin system, responsible for maintaining homeostasis and governing several critical processes, has been considered the main system involved in the pathogenesis and progression of COVID-19. Here, we aimed to assess the possible association between variants in the RAS-related genes and COVID-19 susceptibility and severity in a sample of the Moroccan population. A total of 325 individuals were recruited in this study, with 102 outpatients, 105 hospitalized patients, and 118 healthy controls negative for SARS-CoV-2 infection, and subjected to NGS gene panel sequencing containing eleven RAS pathway genes. A total of 65 functional variants were identified, including 63 missenses, 1 splice, and 1 INDEL. Most of them were rare, with 47 (72%) found in a single individual. According to the common disease/common variant hypothesis, five common candidate variants with MAF > 10% were identified (ACE2 rs2285666, TMPRSS2 rs12329760, AGT rs699 genes, ACE rs4341, and ACE rs4343). Statistical analysis showed that the ACE rs4343 AA genotype was associated with a 2.5-fold increased risk of severe COVID-19 (p = 0.026), and the T genotype of the ACE2 rs2285666 variant showed a borderline association with susceptibility to SARS-CoV-2 in males (p = 0.097). In conclusion, our results showed that the RAS pathway genes are highly conserved among Moroccans, and most of the identified variants are rare. Among the common variants, the ACE rs4343 polymorphism would lead to a genetic predisposition for severe COVID-19.

Prevalence of the protective OAS1 rs10774671-G allele against severe COVID-19 in Moroccans: implications for a North African Neanderthal connection.

The clinical presentation of COVID-19 shows high variability among individuals, which is partly due to genetic factors. The OAS1/2/3 cluster has been found to be strongly associated with COVID-19 severity. We examined this locus in the Moroccan population for the occurrence of the critical variant rs10774671 and its respective haplotype blocks. The frequency of single-nucleotide polymorphisms (SNPs) in the cluster of OAS immunity genes in 157 unrelated individuals of Moroccan origin was determined using an in-house exome database. OAS1 exon 6 of 71 SARS-CoV-2-positive individuals with asymptomatic/mild disease and 74 with moderate/severe disease was sequenced by the Sanger method. The genotypic, allelic, and haplotype frequencies of three SNPs were compared between these two groups. Finally, males in our COVID-19 series were genotyped for the Berber-specific marker E-M81. The prevalence of the OAS1 rs10774671-G allele in present-day Moroccans was found to be 40.4%, which is similar to that found in Europeans. However, it was found equally in both the Neanderthal GGG haplotype and the African GAC haplotype, with a frequency of 20% each. These two haplotypes, and hence the rs10774671-G allele, were significantly associated with protection against severe COVID-19 (p = 0.034, p = 0.041, and p = 0.008, respectively). Surprisingly, in men with the Berber-specific uniparental markers, the African haplotype was absent, while the prevalence of the Neanderthal haplotype was similar to that in Europeans. The protective rs10774671-G allele of OAS1 was found only in the Neanderthal haplotype in Berbers, the indigenous people of North Africa, suggesting that this region may have served as a stepping-stone for the passage of hominids to other continents.

Genome sequencing of severe acute respiratory syndrome COVID-19 EF.1 variant strain obtained from a Moroccan patient.

Here, we report the identification and coding-complete genome sequence of a severe acute respiratory syndrome COVID-19 (SARS-CoV-2) strain obtained from a Moroccan patient. The detected strain EF.1 belongs to the BQ1.1 subvariant of the BA.5 Omicron variant.

Germline mutations of B-Raf proto-oncogene and pathological implications in prostate cancer: observational study.

B-Raf proto-oncogene has been found in a variety of neoplasms. BRAF stimulation can promote tumour proliferation through the activation of the MAP/ERK kinase pathway. This study aimed to determine the germline spectra of BRAF and the association with pathological criteria of prostate tumours. Methods: Fifty blood samples from men treated with prostate cancer were analyzed for BRAF germline mutations and confirmed by Sanger sequencing, in addition, to establishing the frequencies and clinical correlations of frequent mutations in the BRAF gene for both exon 11 and exon 15. The frequency and distribution of high-frequency mutations were analyzed according to the pathological criteria of the patients. Results: Frameshift mutations: c.1628_1629insA and c.1624_1625insT with a frequency of (46%) and (18%), respectively, Nonsense mutations: c.1181C>A (p.Ser394Ter) was detected in one patient, missense mutations: c.1226A>G (p.Gln409Arg), c.1270T>C (p.Trp424Arg), c.1270_1271delins2 (p.Trp424Leu), with a frequency of (4%) were detected. There was no significant difference between mutation carriers and non-carriers regarding medical and surgical history, but prostate-specific antigen concentration was significantly different between the two groups. Conclusion: The results of this study elucidate the presence and involvement of germline mutations in prostate cancer, which could serve as a potential indicator for the diagnosis and therapeutic management of prostate cancer in the population studied.

Advanced Non-small Cell Lung Cancer: EGFR Mutation Analysis Using Pyrosequencing and the Fully Automated qPCR-Based IdyllaTM System.

BACKGROUND: Epidermal growth factor receptor (EGFR) mutation status is of a major clinical significance in non-small cell lung cancer (NSCLC) management, as it guides therapeutic decision making to target patients for a better response to therapy. This implicates the introduction of EGFR mutation analysis as the standard of care for Moroccan NSCLC patients, which in itself entails the implementation of targeted methods for routine EGFR mutation analysis in our laboratories. In this study, we aimed to present 2 targeted methods for EGFR mutation identification and to determine the prevalence and spectrum of EGFR mutations in NSCLC Moroccan patients. METHODS: A retrospective investigation of a cohort of 340 patients was undertaken to analyze somatic EGFR mutations in exons 18 to 21 using pyrosequencing and the IdyllaTM system. RESULTS: Of the enrolled patients, 70.9% were males and 29.1% were females. Predominately, 92% of cases had adenocarcinoma, and 53.7% of patients self-reported a history of smoking. Overall, 73 patients (21.7%) harbored an EGFR mutation, the most prevalent of which were the exon 19 deletions (53.4%) followed by exon 21 substitutions (31%). Exon 18 mutations and exon 20 alterations occurred in 8.1% and 6.7% of the positive EGFR mutation cases, respectively. Of the analyzed cases, all of the EGFR-mutated patients had adenocarcinoma. EGFR mutation prevalence was significantly higher in females (females vs males: 38.4% vs 14.5%, P < .001) and non-smokers (non-smokers vs non-smokers: 36% vs 10.3%, P < .001). The featured pyrosequencing and the IdyllaTM system are targeted methods endowed with high sensitivity and specificity as well as other compelling characteristics which make them great options for routine EGFR mutation testing for advanced NSCLC patients. CONCLUSION: These findings underline the imperious need for implementing quick and efficient targeted methods for routine EGFR mutation testing among NSCLC patients, which is particularly useful in determining patients who are more likely to benefit from targeted therapy.

Coding-Complete Genome Sequences of an Omicron Subvariant (BA.5.2.20) of SARS-CoV-2.

Here, we present the complete coding sequences of two severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains that were recovered from a nasopharyngeal swab from a female patient and the second viral passage in cell culture. After testing, both strains were identified as BA.5.2.20, a subvariant of Omicron.

Neutralising antibodies against SARS-CoV-2 give important information on Covid-19 epidemic evolution in Rabat, Morocco, March 2020-February 2021.

Background: The SARS-CoV-2 is an extremely contagious and acute viral disease mainly affecting humans. Objective: To estimate seroprevalence of SARS-CoV-2 neutralizing antibodies (NAbs) for illegible armed force individuals living in Rabat, Morocco. Method: A convenience sample (N = 2662) was conducted from May 2020 to February 2021. We used the standard neutralization assay to quantify the NAbs titers. A serum was positive when the titer was 1:4. High positive NAbs titers were defined when ≥ 1:32. Results: Demographic and socioeconomic status did not affect seroprevalence data. An overall seroprevalence of 24,9% was found. Sera from blood donors, young recruits and auto-immune population had lower NAbs titers. However, titers were above 1:16 in 9% of the population with high risk of SARS-CoV-2 exposure. Seropositivity increased over time with values reaching peaks after the epidemic waves (2.4% in May 2020; 16.2% in August 2020; 22.7% in December 2020 and 37% in February 2021). Conclusion: And increase of NAbs was observed over time and correlated with the post-epidemic waves of COVID-19 in Morocco.

Prevalence and patterns of mutations in RAS/RAF/MEK/ERK/MAPK signaling pathway in colorectal cancer in North Africa.

BACKGROUND: Our review discuss (i) the findings from analyzed data that have examined KRAS, NRAS and BRAF mutations in patients with colorectal cancer (CRC) in North Africa and to compare its prevalence with that shown in other populations and (ii) the possible role of dietary and lifestyle factors with CRC risk.  METHODS: Using electronic databases, a systematic literature search was performed for the KRAS, NRAS, and BRAF mutations in CRC patients from Morocco, Tunisia, Algeria and Lybia.  RESULTS: Seventeen studies were identified through electronic searches with six studies conducted in Morocco, eight in Tunisia, two in Algeria, and one in Libya. A total of 1843 CRC patients were included 576 (31.3%) in Morocco, 641 (34.8%) in Tunisia, 592 (32.1%) in Algeria, and 34 (1.8%) in Libya. Overall, the average age of patients was 52.7 years old. Patients were predominantly male (56.6%). The mutation rates of KRAS, NRAS and BRAF were 46.4%, 3.2% and 3.5% of all patients, respectively. A broad range of reported KRAS mutation frequencies have been reported in North Africa countries. The KRAS mutation frequency was 23.9% to 51% in Morocco, 23.1% to 68.2% in Tunisia, 31.4% to 50% in Algeria, and 38.2% in Libya. The G12D was the most frequently identified KRAS exon 2 mutations (31.6%), followed by G12V (25.4%), G13D (15.5%), G12C (10.2%), G12A (6.9%), and G12S (6.4%). G12R, G13V, G13C and G13R are less than 5%. There are important differences among North Africa countries. In Morocco and Tunisia, there is a higher prevalence of G12D mutation in KRAS exon 2 (≈50%). The most frequently mutation type in KRAS exon 3 was Q61L (40%). A59T and Q61E mutations were also found. In KRAS exon 4, the most common mutation was A146T (50%), followed by K117N (33.3%), A146P (8.3%) and A146V (8.3%). CONCLUSION: KRAS mutated CRC patients in North Africa have been identified with incidence closer to the European figures. Beside established anti-CRC treatment, better understanding of the causality of CRC can be established by combining epidemiology and genetic/epigenetic on CRC etiology. This approach may be able to significantly reduce the burden of CRC in North Africa.

Coding-Complete Sequence of a SARS-CoV-2 Strain from an Omicron (B.1.1.529+BA.1) Variant Detected in Morocco.

Here, we describe the coding-complete sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strain HM36, identified as a strain of concern of B.1.1.529+BA (Omicron).

Coding-Complete Genome Sequences of a Delta Subvariant (AY.33) of SARS-CoV-2 Obtained from Moroccan COVID-19 Patients.

We report here the complete genome sequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains obtained from Moroccan patients with COVID-19. The analysis of these sequences indicates that the identified strains belong to the AY.33 sublineage of the Delta variant.

Prevalence of specific and recurrent/founder pathogenic variants in BRCA genes in breast and ovarian cancer in North Africa.

BACKGROUND: Elucidation of specific and recurrent/founder pathogenic variants (PVs) in BRCA (BRCA1 and BRCA2) genes can make the genetic testing, for breast cancer (BC) and/or ovarian cancer (OC), affordable for developing nations. METHODS: To establish the knowledge about BRCA PVs and to determine the prevalence of the specific and recurrent/founder variants in BRCA genes in BC and/or OC women in North Africa, a systematic review was conducted in Morocco, Algeria, and Tunisia. RESULTS: Search of the databases yielded 25 relevant references, including eleven studies in Morocco, five in Algeria, and nine in Tunisia. Overall, 15 studies investigated both BRCA1 and BRCA2 genes, four studies examined the entire coding region of the BRCA1 gene, and six studies in which the analysis was limited to a few BRCA1 and/or BRCA2 exons. Overall, 76 PVs (44 in BRCA1 and32 in BRCA2) were identified in 196 BC and/or OC patients (129 BRCA1 and 67 BRCA2 carriers). Eighteen of the 76 (23.7%) PVs [10/44 (22.7%) in BRCA1 and 8/32 (25%) in BRCA2] were reported for the first time and considered to be novel PVs. Among those identified as unlikely to be of North African origin, the BRCA1 c.68_69del and BRCA1 c.5266dupC Jewish founder alleles and PVs that have been reported as recurrent/founder variants in European populations (ex: BRCA1 c.181T>G, BRCA1 c1016dupA). The most well characterized PVs are four in BRCA1 gene [c.211dupA (14.7%), c.798_799detTT (14%), c.5266dup (8.5%), c.5309G>T (7.8%), c.3279delC (4.7%)] and one in BRCA2 [c.1310_1313detAAGA (38.9%)]. The c.211dupA and c.5309G>T PVs were identified as specific founder variants in Tunisia and Morocco, accounting for 35.2% (19/54) and 20.4% (10/49) of total established BRCA1 PVs, respectively. c.798_799delTT variant was identified in 14% (18/129) of all BRCA1 North African carriers, suggesting a founder allele. A broad spectrum of recurrent variants including BRCA1 3279delC, BRCA1 c.5266dup and BRCA2 c.1310_1313detAAGA was detected in 42 patients. BRCA1 founder variants explain around 36.4% (47/129) of BC and outnumber BRCA2 founder variants by a ratio of ≈3:1. CONCLUSIONS: Testing BC and/or OC patients for the panel of specific and recurrent/founder PVs might be the most cost-effective molecular diagnosis strategy.

Genome Sequences of the Delta Variant (B.1.617.2) and the Kappa Variant (B.1.617.1) Detected in Morocco.

Here, we report the identification and coding-complete genome sequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains obtained from patients with COVID-19. The strains identified belong to variant of concern B.1.617.2 and variant of interest B.1.617.1.

Complete Genome Sequence of a SARS-CoV-2 Strain Sampled in Morocco in May 2020, Obtained Using Sanger Sequencing.

The complete genome sequence of a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strain was obtained. The strain was isolated from a nasopharyngeal swab specimen from a female patient in Rabat, Morocco, with coronavirus disease 2019 (COVID-19). This strain belongs to clade 20A and has 12 mutations and 8 amino acid substitutions compared to the reference strain Wuhan/Hu-1/2019.

Study of Clinical and Biological Characteristics of Moroccan Covid-19 Patients With and Without Olfactory and/or Gustatory Dysfunction.

BACKGROUND: The epidemic of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), causing coronavirus disease 2019 (COVID-19), presents a significant and urgent threat to global health. This alarming viral infection, declared as pandemic by the WHO in February 2020, has resulted millions of infected patients and thousands of deaths around the world. In Morocco, despite the efforts made by the authorities, the SARS-CoV-2 continues to spread and constitutes a burden of morbidity and mortality. The objective of this study is to describe clinical characteristics of COVID-19 Moroccan patients and to establish the relationship between specific clinical symptoms, namely ageusia and/or anosmia, with these characteristics. METHODS: We performed a descriptive, non-interventional cross-sectional study analyzing data from 108 patients admitted to the VINCI clinic, Casablanca (Morocco). The database includes 39 parameters including epidemiological characteristics, anthropometric measurements and biological analyzes. RESULTS: The average of age of the patients was 43.80 ± 15.75 years with a sex ratio of 1:1. The mean body mass index of the patients was 25.54 ± 4.63 Kg/m2. The majority of patients had, at least, one comorbidity and among 75% symptomatic patients, about 50% had, at least, three symptoms namely, fever (40.7%), cough (39.8%), myalgia (28.7%), and anosmia and/or ageusia (20.4%). From biological analyzes, we noticed lymphopenia and an elevated protein C reactive and lactate dehydrogenases levels in 24.1, 36.1, and 35.2% of patients, respectively. A disturbance in liver function markers was observed in 15.7% of cases. For the other hemostasis parameters, high levels of prothrombin and platelets were reported in 14.6 and 14.8% of patients, respectively. Comparisons related to the presence of anosmia and/or ageusia did not show any difference for demographic and anthropometric characteristics, while a possibility of a significant difference was revealed for certain biological parameters, particularly the levels of lymphocytes, D-dimer and troponin. CONCLUSION: This study provides significant findings that will be used not only to supplement previous studies carried out in Morocco in order to resume the epidemiological situation in comparison with other countries, but also to improve the quality of the diagnosis of COVID-19 patients by identifying all the symptoms of the disease and better understanding its clinical outcomes.

Role of Training and Experience in Biosafety Practices Among Nurses Working in Level 2 or 3 Patient Containment.

Introduction: This study was conducted to assess the difference in biosafety behaviors among nurses working in a levels 2 and 3 hospital unit in Morocco and the need for additional biosafety training before working in a high-containment facility. Methods: A prospective cross-sectional and observational study was conducted among new and experienced nurses in the Center of Virology and Tropical Infectious Diseases of the Military Teaching Hospital Mohammed V of Rabat, Morocco, between December 1, 2017 and February 28, 2018 (3 months). We compared behaviors in biosafety practices (donning gloves, handwashing, etc) between the 2 groups, and data were collected by observing workers directly or via camera. Results: A total of 31 nurses, 14 newly graduated nurses (45.2%) and 17 experienced and trained (54.8%) nurses, were observed during 89 entries into high-containment hospital rooms. Among the behaviors observed, only donning an inner pair of gloves was observed consistently between the 2 groups. Experienced and trained nurses washed their hands (100%) and observed the antiseptic contact time (71%), whereas recently graduated nurses ignored these activities significantly (79% and 32%, respectively). During handling of the patient, the trained and experienced nurses were significantly better than the new ones in the practices of biosafety and personal protection. Gaps in biosafety were observed more frequently among newly graduated workers, especially donning outer gloves before patient contact, nurses touching their faces, and the act of wearing jewelry while working. Conclusion: Safety training should begin early in the training curriculum with theoretical and practical courses. It must be introduced into the general biosafety policy as part of a safety and standard working culture. It is recommended that personnel be required to pass a proficiency exam in critical biosafety tasks before working with high-risk patients.

[Peripheral neuropathy during Infliximab therapy: a case study]. / Neuropathie périphérique sous Infliximab: étude d'une observation.

Anti TNF alpha treatments are wide spectrum therapies. Multiple side effects have been reported in recent years, particularly peripheral neuropathies. We report a case of axonal neuropathy occurring three months after starting treatment with Infliximab. Our study focused on a 60-year old female patient treated for therapy-resistant hemorrhagic rectocolitis, requiring treatment with infliximab. Three months later, the patient had sensory axonal neuropathy. Etiologic assessment remained negative and dose reduction was accompanied by an improvement in symptoms. The time between initiation of treatment with Infliximab and the onset of clinical manifestations as well as improvement after dose reduction advocate the responsibility of infliximab in the occurrence of sensory neuropathy. Its management is not standardized and should be discussed case by case.

[Granulomatous spondylodiscitis: due mainly to tuberculosis but lymphoma cannot be excluded]. / Spondylodiscite granulomateuse: surtout la tuberculose mais ne pas omettre le lymphome.

Lower back pain is due to multiple etiologies that make diagnosis difficult. Primitive spinal lymphoma is rare and its diagnosis often requires ultrasound-guided biopsy. A 30-year old man hospitalized for inflammatory lumbago evolving within the context of an impaired general condition. Phisical examination revealed pain on palpation of the L2-L3 vertebral apophysis without peripheral tumor syndrome. Laboratory tests showed an inflammatory syndrome. Morphological assessment was in favour of a spondylodiscitis. The first biopsy showed granulomatous osteitis.Clinical and radiological worsening during antibacillary treatment led to reconsider the original diagnosis and a second biopsy confirmed the diagnosis of lymphoma. The diagnosis of skeletal tuberculosis in particular spinal tuberculosis requires bacteriological or histological confirmation in order not to overlook a primitive bone lymphoma.

Acquired ichthyosis disclosing non-Hodgkin's malignant lymphoma.

When ichthyosis occurs in adulthood, it may signify internal disease, especially malignant hemopathy. A 49-year-old man was admitted to hospital with diarrhea, fever, weight loss and abdominal pain. Physical examination showed generalized ichthyosis with palmoplantar hyperkeratosis and multiple inguinal lymph nodes. Laboratory investigations showed severe pancytopenia. Ultrasonography and CT scan demonstrated intraabdominal lymph node enlargement. Osteomedullary biopsy and open abdominal lymph node biopsy revealed an anaplastic large cell lymphoma. The lymph node involvement subsequently became worse and more severe, and the patient's general condition deteriorated. He died before receiving chemotherapy.

[Acquired ichthyosis revealing hepatic and lymph node tuberculosis]. / Ichtyose acquise révélatrice d'une tuberculose hépatique et ganglionnaire.

INTRODUCTION: Acquired ichthyosis is an uncommon disease usually associated with different systemic diseases. Its characteristic clinical sign is symmetrical scaling of the skin. We report a case of acquired ichthyosis revealing hepatic and lymph node tuberculosis. CASE: A 41-year-old man was admitted to our hospital with weight loss and generalized acquired ichthyosis. Ultrasonography and computer tomography showed enlargement of abdominal lymph nodes. Lymph node and liver biopsy samples were taken during exploratory laparotomy and revealed multiple lymphoepitheliomas with Langhans-type giant cells and central caseous necrosis. The skin lesions resolved progressively after 5 months of antituberculous therapy. DISCUSSION: Ichthyosis occurring in adulthood is often a sign of internal disease, especially malignant conditions. It has also been associated with autoimmune and infectious diseases, sarcoidosis and drugs. It is only rarely reported among patients with tuberculosis but it is possible that the ichthyosis in this case was due to weight loss and deterioration of the patient's general condition.