Studying the pathogenicity of 26 variants characterized in the first molecular analyses of Egyptian aplastic anemia patients.

BACKGROUND: Aplastic anemia (AA) is a bone marrow disorder characterized by peripheral pancytopenia and marrow hypoplasia which can lead to life-threatening complications. Our objective was to study the telomerase genes (TERT and TERC) variants, explore their relationship to telomere shortening and TERT gene expression, and to identify variants in the MPL gene within Egyptian AA patients. METHODS: Forty AA patients and 40 sex- and age-matched healthy individuals as the control group were studied through sequencing of TERT, TERC, and MPL genes. Quantitative real-time PCR (qRT-PCR) was used for measuring TERT gene expression. Telomere length (TL) was measured using the Quantitative Fluorescence In Situ Hybridization (Q-FISH) technique. In silico analysis was performed for the prediction of the pathogenicity of resultant variants. RESULTS: Sequencing of MPL, TERT, and TERC genes identified 26 variants. Eleven variants were identified in the MPL gene. Three of them are pathogenic: two missense [c.305 G>A, c.1589 C>T] and one splice site [g.9130T>G]. TERT gene sequencing showed thirteen variants, among them, four novel [c.484G>A, c.499G>A, c.512G>A, c.3164C>G] and two previously reported [c.835G>A, c.2031C>T] were predicted to be pathogenic. Two variants were characterized within the TERC gene; n.514A>G and n.463 C>T. TERT gene expression was downregulated in 70% of studied patients and the Q-FISH technique detected telomere shortening in 82.5% of patients. CONCLUSIONS: Twenty-six pathogenic and benign variants within the TERC, TERT, and MPL genes were identified among the studied AA patients that were in several cases associated with shortened telomeres and/or lower TERT gene expression. Genotype/phenotype correlation in AA patients is of great importance in explaining the disease severity and guiding therapeutic decisions.

A Review of Chitosan and Chitosan Nanofiber: Preparation, Characterization, and Its Potential Applications.

Chitosan is produced by deacetylating the abundant natural chitin polymer. It has been employed in a variety of applications due to its unique solubility as well as its chemical and biological properties. In addition to being biodegradable and biocompatible, it also possesses a lot of reactive amino side groups that allow for chemical modification and the creation of a wide range of useful derivatives. The physical and chemical characteristics of chitosan, as well as how it is used in the food, environmental, and medical industries, have all been covered in a number of academic publications. Chitosan offers a wide range of possibilities in environmentally friendly textile processes because of its superior absorption and biological characteristics. Chitosan has the ability to give textile fibers and fabrics antibacterial, antiviral, anti-odor, and other biological functions. One of the most well-known and frequently used methods to create nanofibers is electrospinning. This technique is adaptable and effective for creating continuous nanofibers. In the field of biomaterials, new materials include nanofibers made of chitosan. Numerous medications, including antibiotics, chemotherapeutic agents, proteins, and analgesics for inflammatory pain, have been successfully loaded onto electro-spun nanofibers, according to recent investigations. Chitosan nanofibers have several exceptional qualities that make them ideal for use in important pharmaceutical applications, such as tissue engineering, drug delivery systems, wound dressing, and enzyme immobilization. The preparation of chitosan nanofibers, followed by a discussion of the biocompatibility and degradation of chitosan nanofibers, followed by a description of how to load the drug into the nanofibers, are the first issues highlighted by this review of chitosan nanofibers in drug delivery applications. The main uses of chitosan nanofibers in drug delivery systems will be discussed last.

MBL2 gene variants and susceptibility to meningitis in Egyptian patients.

BACKGROUND: Meningitis is inflammation of the membranes enclosing the brain and spinal cord. It is a fatal disease with severe morbidity and mortality. Mannose binding lectin (MBL) encoded by MBL2 gene activates complement system through lectin pathway in innate immunity to defense against the infections. OBJECTIVE: The current study aimed to investigate the promoter and exon 1 variants of MBL2 gene among Egyptian patients having meningitis to explore their role in disease susceptibility. PATIENTS AND METHODS: This case-control study, included 53 patients and 50 sex and age matched controls. MBL2 genotyping was done using Sanger sequencing. RESULTS: The frequency of one promoter (c.-290C > G) and four in exon 1 (c.161G > A, c.170G > A, c.154C > T and c.132C > T) as well as another one located in its 5'utranslated part (c.-66C > T) variants were estimated. The incidence of the four individual exonic variants was not significantly different between cases and healthy individuals (all P > 0.05). The promoter variant, c.-290C > G was found in all examined patients (84.9% of the patients in homozygote state and 15.1% of patients in heterozygous state) with a highly significant variance in the prevalence of this variant between cases and control group (p = 0.0001). Additionally, UTR variant (c.-66C > T) was also significantly higher in patients than controls (P = 0.033).In comparison with clinical outcome, it was found that c.170G > A variant named C allele was associated with favorable outcome in the studied patients (P = 0.025). CONCLUSION: The results obtained showed that the Promoter (c.-290C > G) and UTR (c.-66C > T) variants of MBL2 gene may be potential risk factors for disease susceptibility in Egyptian cases with meningitis. Our results also proposed that c.170G > A (C allele and CC genotype) could affect the severity and play a protective role in these patients. The other genetic variants of MBL2 gene, including c.132C > T, c.161G > A (A > B), and c.154C > T (A > D) that were investigated, did not show any association with susceptibility or severity of meningitis.

Enhancement of Antimicrobial and Dyeing Properties of Cellulosic Fabrics via Chitosan Nanoparticles.

The primary goal of this study is to prepare chitosan nanoparticles (CSNPs) by the ionic gelation method via the treatment of chitosan (0.2 wt.%) with tripolyphosphate (0.2 wt.%) ultrasonically for 45 min. FT-IR spectroscopy and TEM images were used to characterize and validate CSNP production. Cellulosic materials with different concentrations of CSNPs have better antibacterial and colouring characteristics. The treated cellulosic fabrics were analyzed by FT-IR spectroscopy, SEM, and thermogravimetric analysis. Colourimetric data measurements expressed in K/S values were used to evaluate the impact of CSNPs on the dyeing affinity of cellulosic materials. In addition, antibacterial activity against bacteria and fungi was tested on the treated cellulosic fabrics. According to the K/S values, cellulosic textiles treated with CSNPs (0.3 wt.%) had a better affinity for acid dyeing. These textiles also offer better antibacterial properties and are more resistant to washing, light, and rubbing. A cytotoxicity study found that CSNPs give cellulosic materials antibacterial and acid dyeing properties, which is good for the environment.

Narrative role of vitamin D receptor with osteoporosis and obesity in a sample of Egyptian females: a pilot study.

BACKGROUND: Vitamin D receptor (VDR) is known as one of the cellular regulators for several metabolic pathways indicating its pivotal role in the pathological pathway of numerous diseases. Considering the high frequency of osteoporosis and obesity among women, the current study aimed to explore the prospective assembly of the most frequent two VDR loci, single nucleotide polymorphism SNPs rs731236 (TaqI) and rs7975232 (ApaI) with a genetic predisposition to osteoporosis (skeletal) and obesity (chronic non-skeletal disorders), in Egyptian women. This was a cross-sectional study, including 97 Egyptian females (25-65 years), randomly chosen, from all employees and workers of the National Research Centre, Egypt. Anthropometric measurements (weight, height, BMI), dual-energy X-ray absorptiometry (DEXA), and molecular genetic analysis were done. RESULTS: The variation of ApaI genotype between the normal and osteoporotic groups denotes a remarkable association of the homozygote ApaI genotype with osteoporosis risk. Among the normal weight group, bone mineral density (BMD) was significantly associated with TaqI VDR gene polymorphism as the presence of the heterozygote genotype was accompanied with higher BMD while the homozygote one was detected with lower BMD. Also, TaqI VDR gene polymorphism was significantly associated with BMI when participants were divided according to the presence of osteoporosis; increased BMI was expressed in the non-osteoporotic women group carrying the homozygote genotype of Taq I VDR gene while the presence of the heterozygote genotype (TaqI) in the osteoporotic group was associated with increased BMI. CONCLUSIONS: The heterozygote TaqI genotype is protective against the osteoporosis phenotype and accompanied with increased BMI among osteoporotic women, while the homozygote ApaI genotype has a significant association with osteoporosis risk.

Defining the molecular pathology and consequent phenotypes in Egyptian HB patients.

BACKGROUND: Hemophilia B (HB) (also known as Christmas disease) is a rare X-linked recessive disorder characterized by spontaneous or prolonged hemorrhages caused by mutations in Factor 9 (F9) gene leading to deficient or defective coagulation F9. Our study aimed at identifying the causative mutations within a sample of HB Egyptian patients. The present study comprised clinical data of eleven HB patients descending from six unrelated families and a seventh family including a carrier mother with a history of deceased HB sibling. Sequencing of F9 gene was performed. RESULTS: The study revealed four mutations; two missense NM_000133.3:c.676C>G, (P.Arg226Gly) and NM_000133.3:c.1305T>G, (p.Cys435Trp), and two nonsense mutations NM_000133.3:c.880C>T, (p.Arg294*) and NM_000133.3:c.1150C>T, (p.Arg384*), identified mutations spanned exons 6 and 8 of which a total of three mutations are located in hotspot exon 8 of F9 gene. CONCLUSIONS: Reviewing the literature, this is the first molecular analysis of F9 gene in HB Egyptian patients. Consistent genotype/phenotypic severity correlation could be concluded, helping proper genetic counseling and prenatal decision taking.

Effect of Environmental Contaminants on Antioxidant Defense System in Fringe-Lip Mullet (Crenimugil crenilabis) from Suez Canal, Egypt.

Fish are widely used as model organisms for the assessment of the quality of aquatic environment and can therefore serve as bio-indicators of environmental contamination. The current research investigates the eco-physiological damage to fishes in Suez Canal for enhancing the biomonitoring of this area through the biochemical investigation, were estimated in the gills, kidney and liver of Mullet, (Crenimugil crenilabis) from Suez Canal, Egypt. This area gets a lot of wastes discharged from several industries, and it is considered as the main routes of many tankers. Crenimugil crenilabis weigh up approximately 350-600 g were fished from Nabq Managed Resource Protected Area as a control area and Suez Canal as a polluted area. Compared with control, a significant changing in the biochemical analysis results, shows that there was an initiation of oxidative stress in the tissue of Mullet from the Suez Canal which indicating the contamination status of this area.

Urtica dioica extracts abolish scopolamine-induced neuropathies in rats.

Alzheimer's disease (AD) is characterized by alterations in monoamines, oxidative stress, and metabolic dysfunctions. We aim to assess the therapeutic impacts of roots or leaf extract from Urtica dioica (UD; stinging nettle) against scopolamine (SCOP)-induced memory dysfunction, amnesia, and oxidative stress in rats. Spatial memory was assessed by Y maze test. Tissue analyses of norepinephrine (NE), dopamine (DA), serotonin (5-HT), malondialdehyde (MDA), nitric oxide (NO), glutathione (GSH, GSSG), AMP, ADP, and ATP were assessed by HPLC. mRNA levels of Tau and Hsp70 were estimated by PCR. UD extracts particularly nettle root (NR) significantly normalized the SCOP-induced memory deficits even more potent than sermion (SR) and donepezil (DON). Similarly, NR had potent therapeutic impacts on the levels of cortical and hippocampal monoamines e.g. DA, NE, and 5-HT. SCOP induced a dramatic oxidative stress as measured by MDA, NO, and GSSG levels; however, UD extracts showed significant anti-oxidative stress impacts. Additionally, UD extracts restored ATP levels and reduced the levels of AMP and ADP compared to SCOP-treated rats. Furthermore, cortical Tau and hippocampal Hsp70 were modulated by UD extracts particularly NR compared to the SCOP group. In conclusion, UD extracts particularly roots have potential therapeutic impacts against SCOP-induced neuroinflammatory and/or Alzheimer-like phenotype in rats.

Genomic alterations in the F8 gene correlating with severe hemophilia A in Egyptian patients.

BACKGROUND: Hemophilia A (HA) is an inherited X-linked recessive coagulation disorder caused by factor VIII (F8) deficiency. F8 rearrangements involving intron 22 (int22) and intron 1 (int1) account for almost half of severe HA phenotype also a hotspot exon 14 provides numerous mutational patterns. This study aims to identify F8 gene mutations among Egyptian HA patients. METHODS: DNA samples from 60 HA patients were screened for int22 and int1 rearrangements using simplified inverse shifting PCR (IS-PCR) followed by exon 14 sequencing. Also, four uncharacterized patients were studied by targeted exome sequencing. RESULTS: In 33.3% of the studied patients, we identified three int22 rearrangements, three exon 14 mutations (two frameshift; one novel (NM_000132.3:c.2734_2735delAA, p.(N912Ffs*6)), a second reported mutation (NM_000132.3:c.3091_3094delAGAA, p.(K1031Lfs*9)), and one nonsense mutation (NM_000132.3:c.2440C>T, p.(R814*)). All identified mutations were detected in patients with severe HA phenotype. Targeted exome sequencing could not detect any known pathogenic variants. CONCLUSION: Intron 22 rearrangement and exon 14 mutations correlate with most severe hemophilia A Egyptian patients.

Obesity phenotype in relation to gene polymorphism among samples of Egyptian children and their mothers.

Obesity is complex heterogeneous disease controlled by genes, environmental factors, and their interaction. Genetic factors account for 40-90% of the body mass index variations. Body mass index (BMI) of children correlates more closely with maternal than paternal BMI. So, this studu was aimed to investigate the role of leptin receptor LEPR Gln223Arg, the uncoupling protein 2 (UCP2 G 866 A) and insulin receptor gene (INSR exon 17) polymorphisms in the pathogenesis of obesity. A cross-sectional study executed on 130 children and their obese mothers; classified into 2 groups according to their BMI. The 2 groups were evaluated regarding the anthropometry. Restriction fragment length analysis for LEPR Gln223Arg, UCP2 -866 G/A and INSR exon 17 polymorphisms were applied. It was reported that increased risk of obesity was found in LEPR AG + AA genotype and the A allele. Significant statistical difference was detected only in female children. Concerning UCP2, the AG followed by the GG genotype was the most frequent in all groups and the G allele was the mostly present in obese mothers and obese male children but with no statistical significance. There was difference in the INSR genotype and alleles between groups, but this difference was not statistically significant. This study concluded that the LEPR Gln223Arg, UCP2 G 866 A and INSR exon 17 polymorphisms are related to obesity in Egyptian population. Further researches on larger population are recommended to ascertain the implications of LEPR, UCP2 and INSR polymorphisms in obesity.

MTDH and MAP3K1 are direct targets of apoptosis-regulating miRNAs in colorectal carcinoma.

Artificially designed miRNAs mimics and inhibitors that specifically target known oncogenes have attracted significant research attention. Herein, we aimed to explore whether MIR-375, MIR-145, and MIR-224 are involved in induction of apoptosis of CRC cells by regulating apoptosis-mediating genes MTDH, MAP3K1, PDK1, BAX, and BCL-XL. MTT assay was used to assess cell growth. Apoptosis was determined in terms of caspase activity measurement and phosphatidylserine detection using annexin V staining by flow cytometry. Quantitative real time PCR, Western blotting, and luciferase reporter assay were carried out to validate genes regulation and targeting by miRNAs. We found that ectopic expression of MIR-375 and MIR-145, and inhibition of MIR-224 can decrease cell growth and induce cell ability to undergo early apoptosis. At mRNA level, transfected cells displayed down-regulation of MTDH, PDK1 and BCL-XL, while BAX and MAP3K1 were up-regulated. Protein expression of MTDH was decreased in cells transfected with MIR-145 mimic and MIR-224 inhibitor but remained unchanged in MIR-375 mimic-transfected cells. Furthermore, MAP3K1 protein expression exibited a decreased level after MIR-375 transient expression with no significant change after MIR-145 mimic or MIR-224 inhibitor transfection. Luciferase reporter assay revealed that MIR-375 and MIR-145 can bind to 3'UTR of MTDH, supporting that MTDH is directly targeted by both miRNAs. Similarly, MAP3K1 was found to be directly regulated by MIR-375. The study concluded that the expression modulation of tumor suppressors MIR-375 and MIR-145, and oncomiR MIR-224 have the ability to induce apoptosis of CRC cells through regulation of apoptosis mediating genes MTDH, MAP3K1, PDK1, BCL-XL and BAX.

Influence of Interleukin-6 (174G/C) Gene Polymorphism on Obesity in Egyptian Children.

BACKGROUND: Obesity is a multi-factorial chronic disorder. A considerable number of studies have been performed to figure out whether there is an association between obesity and polymorphisms of gene IL-6 (174G/C), but the results are equivocal. AIM: This study aimed to find out whether the IL-6 (174G/C) gene was associated with the risk of developing obesity in Egyptian children. SUBJECTS AND METHODS: The study included 149 children and adolescents with age ranged between 9.5 - 18 years. Eighty-five of them were obese which BMIZ-score is > 2, and sixty-four children with BMIZ-score ≤ 2 served as control group. Serum level of IL-6 and genetic analysis for IL-6 (174G/C) gene polymorphism were done. RESULTS: Obese children had significantly higher serum levels of IL-6 as compared to those of control children (P = 0.003). A high percentage of IL-6 polymorphism GC was found in obese subjects (93.7%), while the control group had a higher percentage of IL-6 polymorphism GG (70.6 %). CONCLUSION: Our study showed that carriers of the C allele for the IL-6 (174G/C) polymorphism have higher BMI. As the G174C polymorphism is likely to affect IL-6 expression and its physiological regulation; consequently this polymorphism may affect adiposity.