Effect of salinity on ccmfn gene RNA editing of mitochondria in wild barley and uncommon types of RNA editing.

The primary function of mitochondria is cellular respiration and energy production. Cytochrome C complex is an essential complex that transports electrons in the respiratory chain between complex III and complex IV. One of this complex's main subunits is CcmFN, which is believed to be crucial for holocytochrome assembly. In wild-type plant Hordeum vulgare subsp. spontaneum, four ccmfn cDNAs are subjected to high salt stress (500 mM salinity), 0 h (or control) (GenBank accession no. ON764850), after 2 h (GenBank accession no. ON7648515), after 12 h (GenBank accession no. ON764852), and after 24 h (GenBank accession no. ON764853) and mtDNA of ccmfn gene (GenBank accession no. ON764854). Using raw data from RNA-seq, 47 sites with nucleotide and amino acid modifications were detected. There were ten different RNA editing types, with most of them are C to U. Unusual editing types in plants have also been found, such as A to C, C to A, A to G, A to U, T to A, T to C, C to G, G to C, and T to G. High levels of editing were observed in control as well as treatments of salinity stress. Amino acid changes were found in 43 sites; nearly all showed hydrophilic to hydrophilic alterations. Only C749 showed regulation under salinity stress.

The first report of RNA U to C or G editing in the mitochondrial NADH dehydrogenase subunit 5 (Nad5) transcript of wild barley.

BACKGROUND: Nad dehydrogenase complex in mtDNA has a significant role in cellular respiration. One of the largest subunits in the complex is subunit 5 (Nad5). METHODS AND RESULTS: Four cDNAs of the Hordeum vulgare subsp. spontaneum nad5 gene have been characterized and subjected to four phases of 0.5 M salinity, at 0 h (control, accession no. MT235236), after 2 h (acc. no. MT235237), after 12 h (acc. no. MT235238) and after 24 h (acc. no. MT235239). Utilizing raw data from RNA-seq, ten RNA editing sites were reported. Seven sites have common editing from C to U in positions (C1490, C1859, C1895, C1900, C1901, C1916, C1918). A rare editing event U to C was detected in two positions (U1650 and U1652) and a novel editing event U to G was for the first time in positions nad5-U231. The highest editing level was shown in 2 and 12 h after salinity exposure. After 24 h, these edits were disrupted, possibly due to the launch of the programed cell death mechanism. However, the RNA editing in positions U1650, U1652 and U231 was fixed at all exposure times. CONCLUSIONS: Although study clarified the role of salinity stress in nad5 RNA editing sites, the main achievements are first report of U to G RNA editing in plants at position U231 and first report of U to C editing in the nad5 gene at U1650 and U1652.

Ethylene responsive transcription factor ERF109 retards PCD and improves salt tolerance in plant.

BACKGROUND: The ultimate goal of this work was to detect the role of transcription factors (TFs) concordantly expressed with genes related to programmed cell death (PCD) during PCD and salt stress. This work was based on the hypothesis that TFs and their driven genes likely co-express under different stimuli. The conserved superfamily ethylene responsive factor (AP2/ERF) draw attention of the present study as it participates in the response to biotic and abiotic stimuli as well as to program cell death (PCD). RESULTS: RNA-Seq analysis was done for tobacco (N. benthamiana) leaves exposed to oxalic acid (OA) at 20 mM for 0, 2, 6, 12 and 24 h to induce PCD. Genes up-regulated after 2 h of OA treatment with known function during PCD were utilized as landmarks to select TFs with concordant expression. Knockdown mutants of these TFs were generated in tobacco via virus induced gene silencing (VIGS) in order to detect their roles during PCD. Based on the results of PCD assay, knockout (KO) T-DNA insertion mutants of Arabidopsis as well as over-expression lines of two selected TFs, namely ERF109 and TFIID5, analogs to those in tobacco, were tested under salt stress (0, 100, 150 and 200 mM NaCl). CONCLUSIONS: Results of knockdown mutant tobacco cells confirmed the influence of these two TFs during PCD. Knockout insertion mutants and over-expression lines indicated the role of ERF109 in conferring salt tolerance in Arabidopsis.

Efficient production of lycopene in Saccharomyces cerevisiae by expression of synthetic crt genes from a plasmid harboring the ADH2 promoter.

Lycopene is an effective antioxidant proposed as a possible treatment for some cancers and other degenerative human conditions. This study aims at generation of a yeast strain (Saccharomyces cerevisiae) of efficient productivity of lycopene by overexpressing synthetic genes derived from crtE, crtB and crtI genes of Erwinia uredovora. These synthetic genes were constructed in accordance with the preferred codon usage in S. cerevisiae but with no changes in amino acid sequences of the gene products. S. cerevisiae cells were transformed with these synthetic crt genes, whose expression was regulated by the ADH2 promoter, which is de-repressed upon glucose depletion. The RT-PCR and Western blotting analyses indicated that the synthetic crt genes were efficiently transcribed and translated in crt-transformed S. cerevisiae cells. The highest level of lycopene in one of the transformed lines was 3.3mglycopene/g dry cell weight, which is higher than the previously reported levels of lycopene in other microorganisms transformed with the three genes. These results suggest the excellence of using the synthetic crt genes and the ADH2 promoter in generation of recombinant S. cerevisiae that produces a high level of lycopene. The level of ergosterol was reversely correlated to that of lycopene in crt-transformed S. cerevisiae cells, suggesting that two pathways for lycopene and ergosterol syntheses compete for the use of farnesyl diphosphate.

Calotropis procera accumulates Uzarigenin and Calotropagenin in response to environmental lighting and drought.

Calotropis procera (C. procera) was evaluated as a pharmaceutically useful plant and for its therapeutic effects in the most significant studies. Uzarigenin and Calotropagenin are significant components of this plant that have pharmacological effects on certain systems, including the digestive, immunological, and focal, and peripheral sensory systems. In this study, pathway genes are extracted from high throughput data acc.no. SRR1554320. Seven critical enzymes are involved in studying the effects of sunlight on the formation of Uzaragenin and Calotropagenin in C. procera before and after irrigation. Molecular identification and NCBI submission of six enzyme genes were successful; HSD (acc.no. OQ091761) for 3ß-hydroxystroid dehydrogenase, OR (acc.no. OQ091762) for 5beta-pregnan oxidoreductase, MO (acc.no. OQ091763) for Pregnan monooxygenase, HOX (acc.no. OQ091764) for Steroid hydroxylase, MAT (acc.no. OQ091765) for Melonyletransferase, UHOX (acc.no. OQ091766) for Uzarigenin hydroxylase. During dawn after irrigation, the Uzargenin pathway showed the highest activity, however midday after irrigation was the lowest. The most period that showed high activity for the Uzargenin pathway was dawn after irrigation, however, midday after irrigation was the lowest. This data is confirmed by chromatography analysis (UPLC) to calculate the accumulation of Uzarigenin and Calotropagenin in different periods.

Promoter Methylation-Regulated Differentially Expressed Genes in Breast Cancer.

Background: Breast cancer is one of the most common malignancies among women. Recent studies revealed that differentially methylated regions (DMRs) are implicated in regulating gene expression. The goal of this research was to determine which genes and pathways are dysregulated in breast cancer when their promoters are methylated in an abnormal way, leading to differential expression. Whole-genome bisulfite sequencing was applied to analyze DMRs for eight peripheral blood samples collected from five Saudi females diagnosed with stages I and II of breast cancer aligned with three normal females. Three of those patients and three normal samples were used to determine differentially expressed genes (DEG) using Illumina platform NovaSeq PE150. Results: Based on ontology (GO) and KEGG pathways, the analysis indicated that DMGs and DEG are closely related to associated processes, such as ubiquitin-protein transferase activity, ubiquitin-mediated proteolysis, and oxidative phosphorylation. The findings indicated a potentially significant association between global hypomethylation and breast cancer in Saudi patients. Our results revealed 81 differentially promoter-methylated and expressed genes. The most significant differentially methylated and expressed genes found in gene ontology (GO) are pumilio RNA binding family member 1 (PUM1) and zinc finger AN1-type containing 2B (ZFAND2B) also known as (AIRAPL). Conclusion: The essential outcomes of this study suggested that aberrant hypermethylation at crucial genes that have significant parts in the molecular pathways of breast cancer could be used as a potential prognostic biomarker for breast cancer.

Innovation of new clinical concepts in the correction of facial deformity of patients with post-traumatic condylar hyperplasia condition.

Post traumatic condylar hyperplasia condition is an increase in the total number of cells due to increased activity, which exist only as long as the activity or the stimulus is applied. When it is removed, the tissue returns to the normal state; however, a secondary structural alteration in the general architecture due to accompanying degeneration may render a complete return to the normal state impossible. Mandibular asymmetry following condylar injury is poorly documented as a cause of facial asymmetry. leFort1 (low-level fracture) osteotomies and bilateral sagittal mandibular osteotomies, which are comprehensive surgical plans, correct the facial deformity together with the occlusion. An innovation of new clinical concepts has been used in the correction of facial deformity in 11 patients with post-traumatic condylar hyperplasia condition (syndrome) by applying a new modified allo plastic material (subperiosteal acrylic implant).

New surgical technique in restoring the chin lost due to trauma by using a subperiosteal problast implant.

Bone grafts are the most common techniques used in maxillofacial surgery in restoring any lost or defective bone. This novel surgical technique utilizes a new modified problast to restore the contour of the defected chin.

Innovation of new surgical technique to contour the missing bone in the face of survivors of traffic and war injuries by new modified Problast painted with a mixture of biocompatible materials.

The bone grafts taken from the patient himself (Auto genius graft) to restore the contour of the lost facial bone lost is the ideal technique on the road to recovery from injuries of war or traffic accidents. This novel surgical procedure utilizes a new modified Problast painted with a mixture of a biocompatible materials to restore the lost parts of the zygomatic bones, due to traffic accidents, shell injuries, or bullet injuries.

Consanguinity and Congenital Heart Disease Susceptibility: Insights into Rare Genetic Variations in Saudi Arabia.

Congenital heart disease (CHD) encompasses a wide range of structural defects of the heart and, in many cases, the factors that predispose an individual to disease are not well understood, highlighting the remarkable complexity of CHD etiology. Evidence of familial aggregation of CHD has been demonstrated in different communities and for different cardiac lesions. Consanguinity, particularly among first cousins, is an added risk factor for these families, particularly in societies where it is considered a common cultural practice, as confirmed in previous studies conducted in Saudi Arabia and other countries. Through comprehensive genetic testing of affected families, we have been able to better understand the genetic basis of the various cardiac lesions and to delineate the molecular mechanisms involved in cardiac morphogenesis. In this review, we discuss the epidemiology and genetics of CHD in consanguineous populations focusing on Saudi Arabia as an extensive study model to address current advances and challenges in the clinical genetic diagnosis and prevention of CHD.

First report of triterpenes pathway in Calotropis procera revealed to accumulate beta-amyrin.

The major reports on Calotropis procera (C. procera) indicated the importance of this plant as a resource of pharmaceutically active ingredients as well as its medical advantages. ß-amyrin (BA) is a significant substance in this plant and has a pharmacological effects in some frameworks, like focal and fringe sensory system, digestive and immune systems. In this study, the impact of sunlight before and after irrigation on the BA production in C. procera is studied its pathway with involved eight key enzymes. The eight enzymes' genes were characterized and successfully submitted to NCBI; AAS (acc.no. KU997645) for α-amyrin synthase, BAS (acc.no. MW976955) for ß-amyrin synthase, SE (acc.no. MW976956) for squalene epoxidase, SS (acc.no. MW976957) for squalene synthase, GPPS, (acc.no. MW976958) for geranyl pyrophosphate synthase, FPPS (acc.no. MW976959) for farnasyl pyrophosphate synthase, CAS1, (acc.no. MZ00598) for cycloartenol synthase1 and LS (acc.no. MZ005982) for lupeol synthase. qRT-PCR analysis revealed high expression levels of GPPS, FPPS, SS, SE, and BAS genes at all times specially midday. Otherwise, CAS1, LS and BAS expression levels were very low at all daylight periods. The UPLC ß-amyrin data are in accordance with qRT-PCR results. This indicates that triterpenes biosynthetic pathway in C. procera is going to ß-amyrin accumulation with the highest level at midday.

Complex Inheritance of Rare Missense Variants in PAK2, TAP2, and PLCL1 Genes in a Consanguineous Arab Family With Multiple Autoimmune Diseases Including Celiac Disease.

Background: Autoimmune diseases (AIDs) share a common molecular etiology and often present overlapping clinical presentations. Thus, this study aims to explore the complex molecular basis of AID by whole exome sequencing and computational biology analysis. Methods: Molecular screening of the consanguineous AID family and the computational biology characterization of the potential variants were performed. The potential variants were searched against the exome data of 100 healthy individuals and 30 celiac disease patients. Result: A complex inheritance pattern of PAK2 (V43A), TAP2 (F468Y), and PLCL1 (V473I) genetic variants was observed in the three probands of the AID family. The PAK2 variant (V43A) is a novel one, but TAP2 (F468Y) and PLCL1 (V473I) variants are extremely rare in local Arab (SGHP and GME) and global (gnomAD) databases. All these variants were localized in functional domains, except for the PAK2 variant (V43A) and were predicted to alter the structural (secondary structure elements, folding, active site confirmation, stability, and solvent accessibility) and functional (gene expression) features. Therefore, it is reasonable to postulate that the dysregulation of PAK2, TAP2, and PLCL1 genes is likely to elicit autoimmune reactions by altering antigen processing and presentation, T cell receptor signaling, and immunodeficiency pathways. Conclusion: Our findings highlight the importance of exploring the alternate inheritance patterns in families presenting complex autoimmune diseases, where classical genetic models often fail to explain their molecular basis. These findings may have potential implications for developing personalized therapies for complex disease patients.

The effect of cochlear implantation and post-operative rehabilitation on acoustic voice analysis in post-lingual hearing impaired adults.

Post-lingual deaf adults can develop some vocal abnormalities similar to those developed in pre-lingual deaf individuals. The aim of this work was to study the effect of cochlear implantation followed by post-operative rehabilitation on voice acoustics in post-lingual hearing impaired adults with different durations of hearing loss. The study included 35 post-lingual hearing impaired adults who underwent cochlear implantation. Patients were divided into two groups according to the duration of their hearing loss. Each group was further divided into two subgroups according to whether they received auditory rehabilitation or not. Using the Multi-Dimensional Voice Program (MDVP) parameters, comparisons were made between each subgroup of patients and the normal MDVP Saudi database, and between subgroups of patients. Most of the patients in the two groups reported significant improvement in their MDVP results post-implantation. Further, significantly deviant MDVP parameters were reported in the group of patients with longer duration of hearing loss. Patients who received rehabilitation significantly improved more than those who did not. In conclusion, it appears that cochlear implantation improves the auditory control of voice production in post-lingual deaf adults. Also, it is obvious that cochlear implantation at an early stage of hearing loss gives better results on voice control, especially if augmented with auditory rehabilitation.

Dengue and human health: A global scenario of its occurrence, diagnosis and therapeutics.

Dengue is one of the highest and rapidly spreading vector-borne viral diseases with high mortality rates. The infection causes acute febrile illness, a major public health concern in the tropics and subtropics globally. The disease is caused by an RNA virus that belongs to the Flaviviridae family. The virus is transferred to humans by the mosquito vector called Aedvrves aegypti, which is the cause of new prevalent sicknesses worldwide. These vector-borne viral diseases spread very fast and pose public health and economic challenges that deemed various prevention and control techniques. The Flavivirus genus consists of five different types of viruses starting from DENV-1 to DENV-5. Thus, the present review focuses on the origin of the virus, how the Dengue virus can be detected, infection, the morphology of the virus, its classifications as proposed by ICTV, the replication and genome of the dengue virus, translation, receptor binding, and some vaccine trial volunteers. In addition, it highlights the current challenges and limitations of effective dengue treatment.

Surgical and Nonsurgical Lines of Treatment of Reinke's Edema: A Systematic Literature Review.

OBJECTIVES: The aim of this study was to carry out a systematic review of different surgical and nonsurgical lines of treatment of Reinke's edema to identify the effectiveness and the guidelines described to use each of them. METHODS: We searched PubMed and Coherence for randomized controlled trials and case reports studies did on adult humans aged from 18 to 80 years, from January 1997 to December 2018. We included studies that treated patients with Reinke's edema by one and/or combinations of the following interventions: cold steel microlaryngeal phonosurgery, microdebrider, CO2 laser, photoangiolytic laser, voice therapy, Steroid and Hyaluronidase injection. We selected studies that assessed patients pre- and post-treatment by at least one of the following measures: subjective assessment of the patient's voice by the voice handicap index, perceptual analysis of the voice by GRBAS scale, video laryngeoscopic examination of the VFs and the lesion, aerodynamic measures, and acoustic analysis of the voice by the computerized speech lab. We assessed quality of the included studies with the Cochrane risk of bias assessment Tool. RESULTS: After removal of duplicates, research yielded 262 studies. Of 217 abstracts and titles, 36 full-text articles were read, and one study was added through hand search, resulting in 10 included studies. CONCLUSIONS: Most of literature analyzed were deficient to address the effectiveness of any of the six lines included in this study. This is mainly due to the small number of the included articles and the small sample size in most of these studies. We found only 10 articles that reported the effectiveness based on comparing the results of before and after treatment. In addition, the variability of outcome measures used and the lack of the comprehensive assessment of the patient's voice, vocal image in most of the included studies made it hard to us to compare the results of any of the included studies. More researches with larger sample size and accurate randomization are needed for further accurate assessment of the effectiveness of the surgical and nonsurgical lines of treatment of Reinke's edema. The future researches should take in their consideration the use of an agreed comprehensive assessment protocol for assessing and comparing the outcome measures before and after treatment.

Knowledge of and Attitudes Toward the Use of Anabolic-Androgenic Steroids among the Population of Jeddah, Saudi Arabia.

PURPOSE: The aim of this study was to investigate the correlation between the use of anabolic-androgenic steroids (AASs) among the population of Jeddah, Saudi Arabia, and their knowledge and attitudes. METHODS: This was a community-based, cross-sectional observational study. This study was conducted using a questionnaire that was distributed among the population during the period from February 3, 2018, to February 25, 2018. This questionnaire comprised 31 questions, designed to evaluate the knowledge and attitudes toward using AASs. RESULTS: A total of 300 participants were enrolled in the study. The mean age of the population was 30.66 ± 9.2 years. Fourteen participants admitted using AASs, with a percentage of 4.7%, among whom 85.7% were male (P = 0.0005). Seventy-eight percent of AAS users believed that AASs do not cause tolerance when taken for a longtime (P = 0.023). However, the majority of both AAS users and nonusers did not agree on taking AASs for a longtime. Our results showed a strong correlation between not taking AASs and not consuming energy drinks (P = 0.0023). Half of our respondents exhibited poor knowledge regarding the side effects of AASs. The level of knowledge did not correlate with the use of AAS, gender, exercising, or consuming energy drinks. CONCLUSION: The results showed poor knowledge regarding using AASs among the population of Jeddah. Thus, we recommend having a national awareness program in order to prevent the possible side effects of misusing AASs.

Control of ß-sitosterol biosynthesis under light and watering in desert plant Calotropis procera.

Most scientific studies on Calotropis procera refer to the plant as an important source of pharmaceutical compounds and its valuable benefits in medicine. One of the most important substances in this plant is the potential immunostimulant ß-sitosterol (BS) that acts in improving human health. This study focused on the effects of lighting before and after irrigation on the BS accumulation pathway namely steroid biosynthesis. Studying the enzymes in BS biosynthetic pathway indicated the upregulation at dawn and predusk of the SMT2 and SMO2 genes encoding sterol methyltransferase 2 and methylsterol monooxygenase, two key enzymes in BS accumulation in C. procera. The results almost indicated no regulation at the different time points of the CYP710A gene encoding sterol 22-desaturase, an enzyme that acts in depleting ß-sitosterol towards the biosynthesis of stigmasterol. RNA-Seq data was validated via quantitative RT-PCR and results were positive. The data of ultra-performance liquid chromatography-tandem mass spectrometry analysis with regard to BS accumulation also aligned with those of RNA-Seq analysis. We focused on the effects of light before and after watering on BS accumulation in C. procera. Our results show that BS accumulation is high at dawn in both dehydrated and well-watered condition. While, the BS was dramatically decrease at midday in well-watered plants. This increase/decrease in BS content is correlated with rates of expression of SMT 2 gene. This gene is a key convertor between the different branches in the cardiac glycoside biosynthesis. Accordingly, it could be suggested that BS (or one of the descendent product) may play an important role in C. procera tolerance to drought/light intensity conditions.

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) in Dromedary Camels in Africa and Middle East.

: Dromedary camels are the natural reservoirs of the Middle East respiratory syndrome coronavirus (MERS-CoV). Camels are mostly bred in East African countries then exported into Africa and Middle East for consumption. To understand the distribution of MERS-CoV among camels in North Africa and the Middle East, we conducted surveillance in Egypt, Senegal, Tunisia, Uganda, Jordan, Saudi Arabia, and Iraq. We also performed longitudinal studies of three camel herds in Egypt and Jordan to elucidate MERS-CoV infection and transmission. Between 2016 and 2018, a total of 4027 nasal swabs and 3267 serum samples were collected from all countries. Real- time PCR revealed that MERS-CoV RNA was detected in nasal swab samples from Egypt, Senegal, Tunisia, and Saudi Arabia. Microneutralization assay showed that antibodies were detected in all countries. Positive PCR samples were partially sequenced, and a phylogenetic tree was built. The tree suggested that all sequences are of clade C and sequences from camels in Egypt formed a separate group from previously published sequences. Longitudinal studies showed high seroprevalence in adult camels. These results indicate the widespread distribution of the virus in camels. A systematic active surveillance and longitudinal studies for MERS-CoV are needed to understand the epidemiology of the disease and dynamics of viral infection.

Accent method of voice therapy for treatment of severe muscle tension dysphonia.

Nonorganic dysphonia may present a challenging diagnosis, and management. Here, we present a severe form of nonorganic dysphonia, which we termed as arytenoidal dysphonia. It was a severe form of muscle tension dysphonia, which was described earlier in literature although with different nomenclature. The outcome of the accent method of voice therapy was also presented. We concluded that accent method of voice therapy is proven to be an effective treatment modality of arytenoidal dystonia.

Multifunctional activities of ERF109 as affected by salt stress in Arabidopsis.

Transcriptomic analysis was conducted in leaves of Arabidopsis T-DNA insertion ERF109-knocked out (KO) mutant or plants overexpressing (OE) the gene to detect its role in driving expression of programmed cell death- (PCD-) or growth-related genes under high salt (200 mM NaCl) stress. The analysis yielded ~22-24 million reads, of which 90% mapped to the Arabidopsis reference nuclear genome. Hierarchical cluster analysis of gene expression and principal component analysis (PCA) successfully separated transcriptomes of the two stress time points. Analysis indicated the occurrence of 65 clusters of gene expression with transcripts of four clusters differed at the genotype (e.g., WT (wild type), KO ERF109 or OE ERF109 ) level. Regulated transcripts involved DIAP1-like gene encoding a death-associated inhibitor of reactive oxygen species (ROS). Other ERF109-regulated transcripts belong to gene families encoding ROS scavenging enzymes and a large number of genes participating in three consecutive pathways, e.g., phenylalanine, tyrosine and tryptophan biosynthesis, tryptophan metabolism and plant hormone signal transduction. We investigated the possibility that ERF109 acts as a "master switch" mediator of a cascade of consecutive events across these three pathways initially by driving expression of ASA1 and YUC2 genes and possibly driving GST, IGPS and LAX2 genes. Action of downstream auxin-regulator, auxin-responsive as well as auxin carrier genes promotes plant cell growth under adverse conditions.