Dissection of two drug-targeted regions of Hepatitis C virus subtype 4a infecting Egyptian patients.

Recently, treatment of HCV infection has been improved after the development of direct acting antivirals (DAAs) which target different viral proteins (NS3-4A, NS5A and NS5B). The activity and effectiveness of these DAAs are affected by the presence of resistance associated substitutions (RASs). This study aimed to characterize HCV genotypes circulating among Egyptian HCV patients, to dissect the full sequences of HCV NS3-4A and NS5B regions, and to characterize RASs associated with NS3-4A and NS5B inhibitors in HCV treatment-naïve patients. Genotyping of 80 HCV samples from treatment-naïve patients was done using restriction fragment length polymorphism and phylogenetic analysis based on some full NS5B sequences. Results showed the prevalence of HCV subtype 4a. Twenty four new full sequences of NS3-4A and NS5B regions of subtype 4a were deposited in the GenBank database. In general, the substitutions associated with NS3-4A-targeting drugs were absent predicting possible responsiveness of Egyptian HCV patients to these drugs. In addition, the absence of amino acid substitutions associated with resistance to Sofosbuvir may predict good response to treatment with Sofosbuvir. Some amino acid substitutions associated with resistance to different classes of non-nucleoside inhibitors were detected. Further investigations on treated Egyptian HCV patients may evaluate the effectiveness of the massively used drugs. Many predicted T-cell-binding epitopes in NS3-4A and NS5B regions were found to be highly conserved in the currently studied isolates; a finding that might be important for HCV vaccine development. We demonstrated potential NS3 epitopes that could be used in engineering T cells against HCV epitopes.

A comprehensive evaluation and first molecular report of Theileria ovis infection in small ruminants in Saudi Arabia.

A total of 1000 clinically healthy small ruminants comprising 500 sheep and 500 goats from five districts within Riyadh Province in Saudi Arabia were investigated by routine Giemsa staining for hematozoan parasites. Out of these, 100 sheep and 95 goat samples were investigated by PCR using three pairs of hemoprotozoan-specific primers. Based on microscopic examination, 33.2% of sheep and 25.2% of goats were found infected with hemoprotozoan parasites, while PCR detected hematozoan infection in 46% of sheep and 33.7% of goats. Extensive molecular characterization of hematozoan infection using six pairs of species-specific primers revealed the dominance of Theileria ovis, rather than any other species, which is recorded for the first time in small ruminants in Saudi Arabia. Prevalence of T. ovis in sheep and goats was found to be the highest in Riyadh (32, 48%) followed by AL-Kharj (31, 35%), Ad-Dawadimi (31, 33%), AL-Majmaah (15, 27%), and Rumah (17, 23%). The highest parasite prevalence was recorded in the 3 years of age and > 4 years of age ruminants, while the lowest prevalence was recorded in < 1 year of age ruminants. No noticeable differences in parasite prevalence between male or female ruminants were recorded. Partial sequencing of 18S rRNA gene revealed the infection of the studied ruminants with four new isolates of T. ovis. Further characterization of the pathogenicity and the clinical effects of these T. ovis isolates in sheep and goats is highly needed. The current results can be helpful in protecting and improving livestock industry in the countries that depend on a high number of small ruminants.

Protozoan communities serve as a strong indicator of water quality in the Nile River.

The relationship between the protozoan communities and environmental variables was studied in the Nile River to evaluate their potential as water quality indicators. Protozoans were sampled monthly at six sampling sites in the Nile's Damietta Branch across a spatial gradient of environmental conditions during a 1-year cycle (February 2016-January 2017). The Protozoa community was comprised of 54 species belonging to six main heterotrophic Protozoa phyla. The abundance (average, 1089 ± 576.18 individuals L-1) and biomass (average, 86.60 ± 106.13 µg L-1) were comparable between sites. Ciliates comprised the majority of protozoan species richness (30 species), abundance (79.72%), and biomass (82.90%). Cluster analysis resulted in the distribution of protozoan species into three groups, with the most dominant species being the omnivorous ciliate Paradileptus elephantinus. Aluminium, fluoride, and turbidity negatively affected abundance and biomass, while dissolved oxygen and potassium positively impacted biomass. Of the dominant species recorded over the study area, the amoebozoa Centropyxis aculeata was associated with runoff variables, while the bacterivorous ciliates Colpidium colpoda, Glaucoma scintillans, and Vorticella convallaria were related to the abundance of heterotrophic bacteria, phytoplankton biomass, and total organic carbon. Total dissolved salts, PO4, NH3, NO2, dissolved oxygen, and total organic carbon were the strongest causative factors for protozoa distribution. The α-Mesosaprobic environment at site VI confirmed a high load of agricultural runoffs compared to other sites. This study demonstrates that protozoans can be a potential bioindicator of water quality status in this subtropical freshwater river system.

Trafficking of Plasmodium falciparum chimeric rhoptry protein with Brefeldin A.

Trafficking of the rhoptry chimeric protein RhopH2-GFP, which contains RhopH2 signal peptide plus the downstream five amino acids, was dissected by treating parasites with Brefeldin A at three different time points. Twenty eight hrs-stage trophozoites accumulated the chimera within the parasite endoplasmic reticulum. In 32 hrs-stage schizonts, the chimera was distributed in the parasite cytoplasm but not in the parasitophorous vacuole. In 36 hrs stage-schizonts, the chimera was detected in individual structures around the developing merozoites and, in contrary to non-treated parasites, no immature rhoptry vesicles could be detected in the cytoplasm of immature merozoites. These data show that this chimera is trafficked to the rhoptries via Brefeldin A-sensitive pathway indicating that this trafficking is similar to that of the endogenous rhoptry proteins, and that the five amino acids downstream of the signal peptide cleavage site may contain the sorting signal required for rhoptry targeting.

Evaluation of optical coherence tomography angiography findings in patients with multiple sclerosis.

Purpose: To evaluate optical coherence tomography angiography findings in patients with multiple sclerosis (MS). Methods: This prospective noninterventional study was conducted on 30 eyes of relapsing-remitting MS patients. Group (1) included 10 eyes with a history of optic neuritis (ON), group (2) included 10 eyes without any history of optic neuritis (MS-ON), and group (3) included 10 eyes of normal age/sex/refraction matched participants. Optical coherence tomography (OCT) and OCT-A (ZEISS Cirrus™ HD-OCT Model 4000 (Carl Zeiss-Meditec, Dublin, CA) of the optic disc were done for all patients. Results: The best-corrected visual acuity was diminished in MS cases, especially in patients with ON with P value <0.001. The retinal nerve fiber layer (RNFL) thickness showed a significant decrease in the average thickness and in all quadrants, notably the temporal quadrant in group 1 (P < 0.001). Ganglion cell layer thickness was diminished in average thickness and in all quadrants in both groups of MS, but only the first group showed statistical significance with P value <0.001). In respect to optic disc perfusion, Average, superficial, and deep vascular density index (AVDI, VDI 1, VDI 2) were statistically significantly lower in groups 1, 2 with (P-value < 0.001). Conclusion: Decreased vascular perfusion of the optic nerve in MS patients, especially in those with ON is strongly correlated with the damage of RNFL and ganglion cell layer detected by OCT.

Vision adaptation in the laughing dove (Streptopelia senegalensis, Linnaeus, 1766) inferred from structural, ultrastructural, and genetic characterization.

Neuroanatomy of the retina reflects adaptation and acclimation for dark and light conditions. Retinal cells and genes must be functionally adjusted to various environmental luminosities. Opsins are brilliant molecules to assess the adaptations at the genetic and phenotypic levels. Photic adaptations may reveal new mechanisms that enhance vision abilities. Through the investigation of histological, ultrastructural constituents of the whole retinal layers, and the sequencing of shortwave length sensitive opsin 1 (SWS1) of the laughing dove (Streptopelia senegalensis), the current study confirms the plasticity of the retina in response to the natural photic conditions. Retinal pigmented epithelium has plentiful melanosomes, signifying a highly adapted eye for maximum light perception and protection. Variously colored oil droplets signify high color vision ability. Stratified outer nuclear layer with many Müller cells suggests high sensitivity to dim conditions and high retinal regeneration. The penetration of outer limiting membrane by photoreceptor nuclei and Müller cell microvilli could minimize the light reflection. Oligodendrocytes establish thick myelination demanded for a keen visual acuity. A functional violet sensitive SWS1 with crucial Ser90 is presumed. Molecular phylogeny showed a secondary loss as color vision was shifted back from ultraviolet (UV) sensitivity to the ancestral avian violet sensitivity, thus improving visual resolution. However, SWS1 has some UV sensitive residues. These findings implicate not only spectral adaptations with high color vision ability and acuity but also photoinduced structural reorganizations. Further studies are needed to assess the secrets between photons and the structural genes.

Comparison of corneal biomechanical changes after LASIK and F-SMILE with CorVis ST.

PURPOSE: To evaluate changes in corneal biomechanics after LASIK and F-SMILE. SETTING: Elite Eye Center, Tanta, Egypt. DESIGN: Prospective, randomized, unmasked, interventional comparative case series. PATIENTS AND METHODS: A total of 60 eyes were equally divided into two groups; group A: received LASIK and group B: received F-SMILE. Pentacam and CorVis ST parameters were compared before and 6 months after procedures. P ⩽ 0.05 was used for significance of results. RESULTS: bIOP decreased by 0.762 ± 1.211 mmHg in group A (p = 0.092), and by 2.52 ± 1.389 mmHg in group B (p < 0.001) and the difference between groups became significant (p = 0.001). A1 increased significantly in group A only (p = 0.036); while A2 decreased insignificantly in both groups. DAR increased significantly in both groups (p < 0.001, p = 0.022), but the difference between groups remained insignificant (p = 0.461). IR increased significantly in group A only (p < 0.001) and the difference between groups became significant (p = 0.026). ARTH decreased in both groups (p = 0.245, p = 0.695) and the difference remained insignificant (p = 0.928). SP-A1 decreased in group A by 8.89 ± 1.38 mmHg/mm (p = 0.637), and by 32.6 ± 4.39 mmHg/mm in group B (p < 0.001) and the postoperative difference between groups was significant (p = 0.013). CBI increased significantly in group A (p < 0.001), but insignificantly in group B (p = 0.098) and postoperative difference between groups was highly significant (p < 0.001). A significant correlation was found between change in CCT and corresponding changes in DAR, IR, and ARTH after surgery in both groups. CONCLUSION: The significant differences between groups postoperatively as regards bIOP (p = 0.001), IR (p = 0.026), SP-A1 (p = 0.013), and CBI (p < 0.001) indicate stiffer corneas after F-SMILE and suggest less influence on corneal biomechanics than LASIK.

Induction of fetal abnormalities and genotoxicity by molybdenum nanoparticles in pregnant female mice and fetuses.

Increasing the uses of molybdenum (Mo) nanoparticles in a wide range of applications including food, industry, and medicine, resulted in increased human exposure and necessitated the study of their toxic effects. However, almost no studies are available on their genotoxic effects, especially on pregnant females and their fetuses. Therefore, this study was undertaken to estimate the possible induction of genotoxicity and fetal abnormalities, especially fetal malformations and skeletal abnormalities by Mo nanoparticle administration in mice. Oral administration of Mo nanoparticles resulted in significant decreases in the maternal body weight, the number and length of fetuses as well as skeletal abnormalities mainly less ossification and less chondrification. Administration of Mo nanoparticles also caused DNA damage induction which elevated the expression levels of p53, the vital gene in maintaining the genomic stability and cell differentiation in both maternal and fetus tissues. Similarly, the expression levels of E-Cad and N-Cad genes that control skeleton development have also been increased in the tissues of female mice administered Mo nanoparticles and their fetuses. Thus, we concluded that oral administration of Mo nanoparticles induced genotoxic effects and fetal abnormalities that necessitated further studies on the possible toxic effects of Mo nanoparticles.

5' UTR and NS5B-based genotyping of hepatitis C virus in patients from Damietta governorate, Egypt.

Chronic hepatitis C virus (HCV) infection is a main health problem in Egypt causing high rates of mortalities. Egypt has the highest HCV prevalence in the world, with specific HCV subtypes epidemic and circulating extensively in the country. Different antiviral therapy protocols have been implemented for treating Egyptian HCV patients. Due to the limited data about HCV in Egypt, this study aimed to genotype HCV strains circulating in the Nile Delta Damietta governorate and to investigate the variation in the nonstructural 5B (NS5B) region targeted by the newly approved antiviral drugs. Thirty HCV samples from treatment-naïve patients were genotyped by restriction fragment length polymorphism. Some samples were genotyped by direct sequencing of their 5' untranslated region (UTR) and NS5B regions. Phylogenetic analysis was also performed on the sequences of their NS5B regions. Fourteen new sequences have been deposited in the GenBank database. Results showed that subtype 4a was prevalent in addition to subtype 1g. None of the previously reported NS5B substitutions were detected in the sequenced isolates from treatment-naïve patients, which may be a good predictor for efficient treatment of HCV Egyptian patients with Sofosbuvir. Further studies on Sofosbuvir treated-HCV Egyptian patients are required to investigate whether any NS5B substitutions can confer resistance to treatment.

Dielectric and thermodynamic study of Hb exploring Moringa Oleifera leaves extract potential against radiation damage.

This work has a major interest of the use of Moringa oleifera leaves extract as protective and protective-therapeutic natural supplement helping the biosystem to stand against irradiation induced damage. Rats were orally administered with Moringa oleifera leaves extract by two modalities. The dielectric properties were measured over the frequency range 106 - 2 × 108 Hz and in the temperature window 283-313 K. The dielectric parameters were used to estimate some thermodynamic state functions and evaluate the energy within the Hb molecule. Information source will be the least invasive way consuming haemoglobin from blood sample. Results indicated that some physical parameters were better indicators than others and different modalities showed variant outcomes. In addition, administration of moringa leaves extract proved to have a significant effect that can be used to reduce radiation damage which may be due to constituents of Moringa.

Biophysical and biochemical roles of Moringa oleifera leaves as radioprotector.

It has been found that medicinal plants have chemical and/or therapeutic effects on different diseases related to oxidative damage. This work investigates the use of ethanolic Moringa oleifera leaves extract; as a protective and/or therapeutic agent against damage induced by high acute dose of ionizing radiation. Also, this study aims to explore the associations of electrical properties (relaxation time and DC conductivity of bone marrow) with biochemical markers (SOD, CAT and GSH) to detect and prognosticate radiation effects. Biophysical and biochemical data revealed that Moringa extract can improve the electrical properties of bone marrow and the antioxidants levels in the blood. They also showed that the feeding of Moringa leaves extract post irradiation is preferred to recover rapidly and continuously from radiation effects.

Vector analysis of astigmatic changes after small-incision lenticule extraction and wavefront-guided laser in situ keratomileusis.

PURPOSE: To evaluate and compare the efficacy of the astigmatic correction after wavefront-guided laser in situ keratomileusis (LASIK) and small-incision lenticule extraction (SMILE) and in myopic eyes. DESIGN: Prospective case series. SETTING: Horus Vision Correction Center and Ellite Vision Correction Center, Alexandria, Egypt. METHODS: Myopic eyes with astigmatism up to 4.00 diopters (D) had wavefront-guided LASIK or small-incision lenticule extraction. Visual and refractive changes were evaluated during a 6-month follow-up. The astigmatic changes were evaluated using the Alpins method. RESULT: The study evaluated 107 eyes (55 patients), 52 eyes having wavefront-guided LASIK and 55 eyes, small-incision lenticule extraction. No statistically significant differences were found in the 6-month postoperative sphere between the 2 groups (P = .652). The postoperative manifest cylinder and spherical equivalent were significantly lower in the wavefront-guided LASIK group (P < .001). The 6-month postoperative cylinder was 0.50 D or less in all eyes in the wavefront-guided LASIK groups and in 79.8% in the small-incision lenticule extraction group (P < .001). Vector analysis showed a significantly higher difference vector (P < .001) and angle of error (P = .021) and a significantly lower correction index (P = .001) in the small-incision lenticule extraction group. The mean magnitude of error was -0.07 ± 0.20 (SD) and -0.20 ± 0.35 in the wavefront-guided LASIK group and small-incision lenticule extraction group, respectively (P = .012). CONCLUSION: Wavefront-guided LASIK and small-incision lenticule extraction were safe and effective for the correction of myopic astigmatism, although there was a trend toward undercorrection with small-incision lenticule extraction.

TNF-α gene polymorphisms and expression.

Tumor necrosis factor alpha (TNF-α) is a proinflammatory cytokine with an important role in the pathogenesis of several diseases. Its encoding gene is located in the short arm of chromosome 6 in the major histocompatibility complex class III region. Most of the TNF-α gene polymorphisms are located in its promoter region and they are thought to affect the susceptibility and/or severity of different human diseases. This review summarizes the data related to the association between TNF-α gene and its receptor polymorphisms, and the development of autoimmune diseases. Among these polymorphisms the -308G/A TNF-α promotor polymorphism has been associated several times with the the development of autoimmune diseases, however some discrepant results have been recorded. The other TNF-α gene polymorphisms had little or no association with autoimmune diseases. Current results about the molecules controlling TNF-α expression are also presented. The discrepancy between different records could be related partly to either the differences in the ethnic origin or number of the studied individuals, or the abundance and activation of other molecules that interact with the TNF-α promotor region or other elements.

Targeting tams-1 gene results in underestimation of Theileria annulata infection in diseased cattle in Egypt.

Tropical theileriosis is considered one of the most economically important tick borne diseases that cause significant mortality and morbidity to livestock. In the context of epidemiological studies on livestock in Egypt, this report investigated the spread of Theileria annulata among diseased farm cows (Bos indicus) over one year. Blood samples collected from 130 cows were investigated by routine staining and 64 samples were investigated by PCR assay using two different probes targeting tams-1 gene. Microscopy revealed the infection of 33.8% of animals with Theileria while PCR detected infection in 51% of animals with one primer pair and the other primer pair detected the infection in 31% of animals. Combined PCR results indicated the infection of 68.8% of animals with T. annulata. Seasonal fluctuation of parasite infection was evident with the highest infection percentage and prevalence recorded during summer based on both microscopy and PCR data. For the first time, the current study reports the presence of two T. annulata isolates based on tams-1 gene partial sequence in Egypt. Targeting polymorphic genes for parasite detection may result in underestimation of infection and target gene diversity has to be considered. The high infection with these pathogens in the clinically ill cows necessitates implementing serious programs to minimize their economic burden on the Egyptian farming industry.