Uncomplicated falciparum malaria among schoolchildren in Bajil district of Hodeidah governorate, west of Yemen: association with anaemia and underweight.

BACKGROUND: Malaria, malnutrition and anaemia are major public health problems in Yemen, with Hodeidah being the most malaria-afflicted governorate. To address the lack of relevant studies, this study was conducted to determine the prevalence of Plasmodium falciparum and its relation to nutritional status and haematological indices among schoolchildren in Bajil district of Hodeidah governorate, west of Yemen. METHODS: A cross-sectional study was conducted among 400 schoolchildren selected randomly from four schools in Bajil district. Data about demographic characteristics, risk factors and anthropometric measurements of age, height and weight were collected. Duplicate thick and thin blood films were prepared, stained with Giemsa and examined microscopically for malaria parasites. The density of P. falciparum asexual stages was estimated on thick films. EDTA-blood samples were examined for the haematological indices of haemoglobin (Hb) and blood cell counts. RESULTS: Plasmodium falciparum was prevalent among 8.0% (32/400) of schoolchildren with a mean parasite density of 244.3 ± 299.3/µL of blood and most infections showing low-level parasitaemia, whereas Plasmodium vivax was detected in one child (0.25%). Residing near water collections was a significant independent predictor of falciparum malaria [adjusted odds ratio (AOR) = 2.6, 95.0% CI 1.20-5.72; p = 0.016] in schoolchildren. Mild anaemia was prevalent among more than half of P. falciparum-infected schoolchildren and significantly associated with falciparum malaria (AOR = 5.8, 95.0% CI 2.39-14.17; p < 0.001), with a mean Hb concentration of 10.7 ± 1.0 g/dL. Although the mean values of the total white blood cells, monocytes and platelets were significantly lower in infected than non-infected schoolchildren, they were within normal ranges. More than half of the children were malnourished, with stunting (39.3%) and underweight (36.0%) being the most prevalent forms of malnutrition; 6.3% of children were wasted. Underweight (AOR = 5.3, 95.0% CI 2.09-13.62; p < 0.001) but not stunting or wasting, was a significant predictor of falciparum malaria among schoolchildren. CONCLUSION: Asymptomatic falciparum malaria is prevalent among schoolchildren in Bajil district of Hodeidah Governorate, with predominance of low parasitaemic infections and significant association with mild anaemia and underweight. Residence near water collection is a significant predictor of infection with falciparum malaria among schoolchildren. Further studies among children with severe malaria and those with high parasite densities are recommended.

Association of mutation and low expression of the CTCF gene with breast cancer progression.

BACKGROUND: CTCF encodes 11-zinc finger protein which is implicated in multiple tumors including the carcinoma of the breast. The Present study investigates the association of CTCF mutations and their expression in breast cancer cases. METHODS: A total of 155 breast cancer and an equal number of adjacent normal tissue samples from 155 breast cancer patients were examined for CTCF mutation(s) by PCR-SSCP and automated DNA sequencing. Immunohistochemistry (IHC) method was used to analyze CTCF expression. Molecular findings were statistically analyzed with various clinicopathological features to identify associations of clinical relevance. RESULTS: Of the total, 16.1% (25/155) cases exhibited mutation in the CTCF gene. Missense mutations Gln > His (G > T) in exon 1 and silent mutations Ser > Ser (C > T) in exon 4 of CTCF gene were analyzed. A significant association was observed between CTCF mutations and some clinicopathological parameters namely menopausal status (p = 0.02) tumor stage (p = 0.03) nodal status (p = 0.03) and ER expression (p = 0.04). Protein expression analysis showed 42.58% samples having low or no expression (+), 38.0% with moderate (++) expression and 19.35% having high (+++) expression for CTCF. A significant association was found between CTCF protein expression and clinicopathological parameters include histological grade (p = 0.04), tumor stage (p = 0.04), nodal status (p = 0.03) and ER status (p = 0.04). CONCLUSIONS: The data suggest that CTCF mutations leading to its inactivation significantly contribute to the progression of breast cancer.

AA genotype of cyclin D1 G870A polymorphism increases breast cancer risk: Findings of a case-control study and meta-analysis.

BACKGROUND: Cyclin D1 (CCND1) polymorphisms, a regulator of the cell cycle progress from G1 to the S phase, may lead to uncontrolled cell proliferation and lack of apoptosis. G870A, a common single-nucleotide polymorphism in CCND1 influences breast cancer risk. However, the association between G870A polymorphism and breast cancer risk is ambiguous so far. MATERIALS AND METHODS: In this case-control study, we analyzed the role of G870A polymorphism with breast cancer risk in Indian women. A meta-analysis of 18 studies was also performed to elucidate this association by increasing statistical power. RESULTS: In our case-control study, significant risk association of the CCND1 G870A AA genotype with breast cancer in total cohort (odds ratio [OR], 2.98; 95% confidence interval [CI], 1.64-5.42; P value, 4.96e-04) and premenopausal women (OR, 3.31; 95% CI, 1.54-7.08; P value, .003) was found. The results of the meta-analysis showed that AA genotype of the CCND1 G870A polymorphism significantly increases breast cancer risk in total pooled data (AA vs GG+GA: OR = 1.20; 95% CI = 1.03 to 1.39; P value, 0.016*) and Caucasian (AA vs GG+GA: OR = 1.22; 95% CI = 0.99 to 1.51; P value, .056*) but not in Asian population. Further, a significant protective association with breast cancer was also found in the GA vs AA comparison model in pooled data (OR = 0.73; 95% CI = 0.58 to 0.92; P value, .007*) as well as in Caucasian subgroup (OR = 0.62; 95% CI = 0.49 to 0.94; P value, .022*). CONCLUSION: CCND1 G870A AA genotype was found associated with breast cancer risk. Future association studies considering the environmental impact on gene expression are required to validate/explore this association.

Bioengineered intravaginal isolate of Lactobacillus plantarum expresses algal lectin scytovirin demonstrating anti-HIV-1 activity.

Efforts to develop preventatives against HIV infection through sexual route have identified, among many, algal lectins as the potent molecules for scaffolding HIV entry inhibition. Algal lectin scytovirin (SVN) from Scytonema varium, a cyanobacterium, has anti-HIV effects with the potential for use in sculpting HIV neutralization. We created a recombinant strain of human vaginal L. plantarum for extracellular expression of recombinant (r)SVN. The rSVN protein containing culture supernatant was analyzed for its binding with HIV-1 gp160, and for inhibiting infection with primary R5 and X4 HIV-1 strains in TZM-bl cells. The rSVN protein extant in recombinant L. plantarum culture supernatant binds to HIV-1 gp160 and reduces the HIV-induced cytopathic effect to nearly 56.67% and 86.47% in R5 and X4 HIV-1 infected TZM-bl cells, respectively. The fortified L. plantarum may be explored for its use as a live virucide in vaginal mucosa of high risk women to prevent HIV entry.

Cytomegalovirus aggravates the autoimmune phenomenon in systemic autoimmune diseases.

BACKGROUND: Human Cytomegalovirus (CMV), because of its ability to extensively manipulate host immunity during active infection, has been suggested to be involved in autoimmunity. However, its influence on T-cells and cytokines in systemic autoimmune diseases like systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) is indistinct. METHODS: We investigated the in-vitro response of T lymphocytes from SLE and SSc patients to CMV antigen. Functional activity of T lymphocytes was determined by estimating Th1 (IL-2 and IFN-γ) and Th2 (IL-4 and IL-10) cytokines. RESULTS: We observed that CMV antigen stimulation in-vitro resulted in significant increase in CD4:CD8 T-cell ratio in peripheral blood mononuclear cells (PBMCs) from SLE and SSc patients; response dominated by CD4+ than CD8+ memory T-cells. SSc T-cell response was differentiated by aberrant increase in CD4+CD25+ T-cells. CMV antigen caused elevation in IL-4 and IFN-γ production in both patient PBMCs, whereas IL-2 was also raised in SLE PBMCs. The development of large pool of memory T-cells and overproduction of IFN-γ may result in flare-up of autoimmunity in these patients. CONCLUSION: Our study provides an insight into the immunopathological potential of CMV-reactive immune cells to develop new potential strategies for targeted therapeutic intervention.

Assessment of the Prevalence and Incidence of COVID-19 in Saudi Arabia.

The global pandemic of coronavirus disease 2019 (COVID 19) is reported to have started in Wuhan City, Hebei Province, China. It has spread rapidly all over the world, including Saudi Arabia, having a severe health emergency. This new virus was named as the 2019 novel coronavirus (2019-nCoV), and now severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) based on previous practice and phylogenetic and taxonomic investigations. SARS-CoV-2 belongs to the family of Coronaviridae, Betacoronavirus, Sarbecovirus subgenus, genome ß. Throughout the COVID 19 pandemic, several strains of SARS-CoV-2 have been recognized around the world. The SARS-CoV-2 variants have caused significant morbidity and mortality worldwide and in Saudi Arabia as well. The rate at which COVID-19 spread across borders and affected countries has highlighted the significance of health care systems to nations and global operations. This review focuses on the origin, epidemiology, pathophysiology, transmission, and the impact of this disease, while highlighting the knowledge about SARS-CoV-2 variants.

Mutational Analysis of Circulating Omicron SARS-CoV-2 Lineages in the Al-Baha Region of Saudi Arabia.

Purpose: Omicron (B.1.1.529) is one of the highly mutated variants of concern of SARS-CoV-2. Lineages of Omicron bear a remarkable degree of mutations leading to enhanced pathogenicity and upward transmission trajectory. Mutating Omicron lineages may trigger a fresh COVID-19 wave at any time in any region. We aimed at the whole-genome sequencing of SARS-CoV-2 to determine variants/subvariants and significant mutations which can foster virus evolution, monitoring of disease spread, and outbreak management. Methods: We used Illumina-NovaSeq 6000 for SARS-CoV-2 genome sequencing, MEGA 10.2 and nextstrain tools for phylogeny; CD-HIT program (version 4.8.1) and MUSCLE program for clustering and alignment. At the same time, UCSF Chimera was employed for protein visualization. Results: Predominant Omicron pango lineages in Al-Baha were BA.5.2/B22 (n=4, 57%), and other lineages were BA.2.12/21L (n=1, 14.28%), BV.1/22B (n=1, 14.28%) and BA.5.2.18/22B (n=1, 14.28%). 22B nextstrain clade was predominant, while only one lineage showed 21L. BA.5.2/22B, BA.5.2/22B harbored a maximum of n=24 mutations in the spike region. Twelve crucial RBD mutations: D405N, R408S, K417N, N440K, L452R, S477N, T478K, E484A, F486V, Q498R, N501Y, and Y505H were identified except the lineage BA.5.2/22B in which F486V mutation was not observed. Critical deletions S106 in membrane protein NSP6, E31in nucleocapsid, and L24 in spike region were observed in all the lineages. Furthermore, we identified common mutations of Omicron variants of SARS-CoV-2 in therapeutic hot spot spike region: T19I, D405N, R408S, K417N, N440K, L452R, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, A653V, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K, D1146D, L452R, F486V, N679K and D796Y. The effect of RBD-targeted mutations on neutralizing (NAbs) binding was considerable. Conclusion: The outcome of this first report on SARS-CoV-2 variants identification and mutation in the Al-Baha region could be used to lay down the policies to manage and impede the regional outbreak of COVID-19 effectively.

Evaluation of Extended-Spectrum Beta-Lactamase Resistance in Uropathogenic Escherichia coli Isolates from Urinary Tract Infection Patients in Al-Baha, Saudi Arabia.

Urinary tract infections (UTIs) caused by extended-spectrum beta-lactamase (ESBL)-producing organisms are prevalent in both outpatient and inpatient settings, representing the most often encountered forms of infection. This research aimed to estimate the prevalence of ESBL-UTIs along with other uropathogens in the adult population and to assess the antibiotic activity against Escherichia coli extended-spectrum beta-lactamase (E. coli ESBL) isolates from patient samples in Al-Baha. A retrospective cross-sectional study included patients who presented to King Fahad Hospital in Al-Baha with clinical suspicion of UTI between 1 January 2019 and 30 September 2022. A total of 4406 urine samples with significant microbial growth were included in the scope of this investigation. A collective count of 1644 incidents of Escherichia coli (E. coli) was observed, wherein E. coli constituted 85% of the cases, while the remaining 15% comprised E. coli ESBL producers. The prevalence of E. coli ESBL was observed to be 64.7% in females and 35.3% in males, with a majority (67%) of the affected individuals being over the age of 50. The incidence of E. coli infections in the outpatient setting was found to be greater than that observed in the inpatient setting. E. coli ESBL were sensitive to colistin, tigecycline, amikacin, meropenem, imipenem, and nitrofurantoin by 100% and 93.3-100%, 95-99.6%, 95-99.06%, and 81-91%, respectively. On the other hand, the most resistant agents for E. coli ESBL were the group of cephalosporins, aztreonam, and ampicillin with 100% resistance, ciprofloxacin with 56-74% resistance, and cotrimoxazole with a 45-53% resistance level. ESBL-resistant E. coli strains are moderately prevalent in community- and hospital-acquired UTIs, especially in females and elderly patients (>50 years).

Study of Streptococcus mutans in Early Biofilms at the Surfaces of Various Dental Composite Resins.

BACKGROUND: Biofilm deposit on the composite restoration is a common phenomenon and bacterial growth follows the deposition. The study aims to evaluate Streptococcus mutans (S. mutans) early biofilm formation on the surfaces of various dental composite resins by using the real-time quantitative polymerase chain reaction (qPCR) technique. MATERIALS AND METHODS: Thirty-two discs, where eight discs were in each group of Filtek Supreme Ultra (FSU; 3M, St. Paul, MN), Clearfil AP-X (APX; Kuraray Noritake Dental Inc., Tokyo, Japan), Beautifil II (BE2; Shofu, Inc., Kyoto, Japan), and Estelite Sigma Quick (ESQ; Tokuyama Dental, Tokyo, Japan), were fabricated and subjected to S. mutans biofilm formation in an oral biofilm reactor for 12 hours. Contact angles (CA) were measured on the freshly fabricated specimen. The attached biofilms underwent fluorescent microscopy (FM). S. mutans from biofilms were analyzed using a qPCR technique. Surface roughness (Sa) measurements were taken before and after biofilm formation. Scanning electron microscopy (SEM), including energy dispersive X-ray spectrometer (EDS) analysis, was also performed for detecting relative elements on biofilms. RESULTS: The study showed that FSU demonstrated the lowest CA while APX presented the highest values. FM revealed that condensed biofilm clusters were most on FSU. The qPCR results indicated the highest S. mutans DNA copies in the biofilm were on FSU while BE2 was the lowest (p < 0.05). Sa test signified that APX was significantly the lowest among all materials while FSU was the highest (p < 0.05). SEM displayed areas with apparently glucan-free S. mutans more on BE2 compared to APX and ESQ, while FSU had the least. Small white particles detected predominantly on the biofilms of BE2 appeared to be Si, Al, and F extruded from the resin. CONCLUSION: Differences in early biofilm formation onto various composite resins are dependent on the differences in material compositions and their surface properties. BE2 showed the lowest quantity of biofilm accumulation compared to other resin composites (APX, ESQ, and FSU). This could be attributed to BE2 proprieties as a giomer and fluoride content.

Human polyomaviruses JCPyV and MCPyV in urothelial cell carcinoma: a single institution experience.

Objective: Urothelial cell carcinoma (UCC) is the most common type of urinary bladder. JCPyV and BKPyV have been detected in the urine and tissue of urothelial cell carcinomas (UCC) in immunocompetent patients. Here, we investigated the presence of several HPyVs in UCC samples using diverse molecular techniques to study the prevalence of HPyVs in UCC. Methods: A large single-institution database of urine cytology specimens (UCS; n = 22.867 UCS) has previously been searched for decoy cells (n = 30), suggesting polyomavirus infection. The available urine sediments and formalin-fixed paraffin-embedded (FFPE) tissue samples of UCC patients were tested for the presence of JCPyV-LTAg expression by immunohistochemistry (IHC) labeled with SV40-LTAg antibody (clone: PAb416) and subsequent PCR followed by sequencing. In addition, the presence of the oncogenic Merkel cell polyomavirus (MCPyV) and the presence of human polyomavirus 6 (HPyV6) and 7 (HPyV7) DNA were tested with DNA PCR or IHC. Results: Of the 30 patients harboring decoy cells, 14 were diagnosed with UCC of the urinary bladder (14/30; 46.6%) before presenting with decoy cells in the urine. The SV40-LTAg IHC was positive in all 14 UCC urine sediments and negative in the FFPE tissues. JCPyV-DNA was identified in all five available UCS and in three FFPE samples of UCC (three of 14; 21.4%). Two UCC cases were positive for MCPyV-DNA (two of 14; 14.3%), and one of them showed protein expression by IHC (one of 14; 7.1%). All specimens were HPyV6 and HPyV7 negative. Conclusion: Our findings show the presence of JCPyV in the urine and UCC of immunocompetent patients. Moreover, MCPyV was detected in two UCC cases. In total, five UCC cases showed the presence of either JCPyV or MCPyV. The evidence here supports the hypothesis that these viruses might sporadically be associated with UCC. Further studies are needed to confirm the relevance of JCPyV or MCPyV as a possible risk factor for UCC development.

In silico epitope-based vaccine design against influenza a neuraminidase protein: Computational analysis established on B- and T-cell epitope predictions.

Objective: Influenza A virus belongs to the most studied virus and its mutant initiates epidemic and pandemics outbreaks. Inoculation is the significant foundation to diminish the risk of infection. To prevent an incidence of influenza from the transmission, various practical approaches require more advancement and progress. More efforts and research must take in front to enhance vaccine efficacy. Methods: The present research emphasizes the development and expansion of a universal vaccine for the influenza virus. Research focuses on vaccine design with high efficacy. In this study, numerous computational approaches were used, covering a wide range of elements and ideas in bioinformatics methodology. Various B and T-cell epitopic peptides derived from the Neuraminidase protein N1 are recognized by these approaches. With the implementation of numerous obtained databases and bioinformatics tools, the different immune framework methods of the conserved sequences of N1 neuraminidase were analyzed. NCBI databases were employed to retrieve amino acid sequences. The antigenic nature of the neuraminidase sequence was achieved by the VaxiJen server and Kolaskar and Tongaonkar method. After screening of various B and T cell epitopes, one efficient peptide each from B cell epitope and T cell epitopes was assessed for their antigenic determinant vaccine efficacy. Identical two B cell epitopes were recognized from the N1 protein when analyzed using B-cell epitope prediction servers. The detailed examination of amino acid sequences for interpretation of B and T cell epitopes was achieved with the help of the ABCPred and Immune Epitope Database. Results: Computational immunology via immunoinformatic study exhibited RPNDKTG as having its high conservancy efficiency and demonstrated as a good antigenic, accessible surface hydrophilic B-cell epitope. Among T cell epitope analysis, YVNISNTNF was selected for being a conserved epitope. T cell epitope was also analyzed for its allergenicity and cytotoxicity evaluation. YVNISNTNF epitope was found to be a non-allergen and not toxic for cells as well. This T-cell epitope with maximum world populace coverages was scrutinized for its association with the HLA-DRB1*0401 molecule. Results from docking simulation analyses showed YVNISNTNF having lower binding energy, the radius of gyration (Rg), RMSD values, and RMSE values which make the protein structure more stable and increase its ability to become an epitopic peptide for influenza virus vaccination. Conclusions: We propose that this epitope analysis may be successfully used as a measurement tool for the robustness of an antigen-antibody reaction between mutant strains in the annual design of the influenza vaccine.

Prevalence of adenoviruses as ocular disease causatives in Saudi Arabia.

Although Human Adenoviruses outbreaks are rare, there still could be a potential chance for those viruses to mutate and spread quickly in human populations with severe public health and socioeconomic consequences. Outbreaks often spread fairly quickly with considerable morbidity/mortality. Saudi Arabia's geopolitical and religious significance bring with it, millions of pilgrims, and tourists yearly. This presents a significant potential for HAdVs epidemics. This review shows that even with the mushrooming serotypes and genotypes, the scholarly knowledge on the nature, structure, transmission, and management of HAdVs is already well-established. Significant research is ongoing on pharmacological interventions, which, presently remain speculative and lacking in effectiveness. This review similarly uncovers a shortage of literature, both recent and dated, on epidemic keratoconjunctivitis in either Saudi Arabia or the Middle East.

Inappropriate Dental Antibiotic Prescriptions: Potential Driver of the Antimicrobial Resistance in Albaha Region, Saudi Arabia.

PURPOSE: The aim of this study was to analyze and evaluate dental antibiotic prescriptions in Albaha Region, Saudi Arabia. PATIENTS AND METHODS: A two-year retrospective cohort study was conducted between September 1, 2017 and September 1, 2019 in children and adults. Data collected from the patients' medical records were analyzed using SPSS. The Z-test with Bonferroni correction and descriptive proportions were utilized to compare several levels of categorical variables. RESULTS: Of the 43,255 dental visits, antibiotics were provided during 12,573 (29.1%). The commonly prescribed antibiotics were amoxicillin and amoxicillin combined with metronidazole (56.3% and 16.9%, respectively). Alarmingly, antibiotics were provided in several conditions for which they are medically neither recommended nor indicated; together, they represented 27.8% of those consultations in which antibiotics were prescribed. Female dentists prescribed more antibiotics than male dentists (30%, P = < 0.000), with male patients receiving more antibiotics than female patients (36%, P = <0.0001). CONCLUSION: Unnecessary prescription of antibiotics was observed in the present study. Improving knowledge and awareness of Saudi dentists on dental antibiotic prescription is warranted.

Intestinal parasitoses and schistosome infections among students with special reference to praziquantel efficacy in patients with schistosomosis in Hajjah governorate, Yemen

The majority of the population in Yemen lives in rural areas and suffers from parasitic infections. Therefore, the present study aimed to determine the prevalence of intestinal parasitoses and schistosome infections among the students enrolled in the primary schools in Hajjah governorate ­ north of Yemen, along with an assessment of praziquantel (PZQ) in the treatment of microscopy-confirmed cases of Schistosoma mansoni and Schistosoma haematobium. For this purpose, 780 samples (320 stool and 460 urine) were examined microscopically. The present study revealed an overall infection rate of 75.3% (241/320) with intestinal parasites and Schistosoma mansoni. The detected parasite species included Entamoeba histolytica (27.8%), Hymenolepis nana (12.2%), Giardia lamblia (9.7%), Entamoeba coli (9.4%), S. mansoni (9.1%), Ascaris lumbricoides (6.9%), Trichuris trichiura (3.1%), Enterobius vermicularis (2.8%) and Ancylostoma duodenale (2.2%). Schistosoma haematobium was prevalent among 1.7% (8/460) of the investigated students. On the other hand, PZQ yielded a cure rate of 75.7% of Schistosoma-infected students when administered at 40 mg/kg body weight. However, a 100% cure rate was achieved when administered at 60 mg/kg body weight. Therefore, the findings of the present study highlight the importance of monitoring PZQ efficacy through large-scale studies in different settings endemic for schistosomosis in the country.

Circulating rotavirus G and P strains post rotavirus vaccination in Eastern Mediterranean Region.

OBJECTIVE: To detect changes in circulating strains of rotavirus in the Eastern Mediterranean Region post rotavirus immunization drive. METHODS: We searched MEDLINE, PubMed, ScienceDirect, and the Cochrane Library and specific database website (Nutrition and Food Sciences) for relevant articles. Our search included websites of a number of relevant organizations in addition to gray literature search. Of the 2198 articles found, we included only 35 studies after excluding irrelevant, ineligible, duplicated, and very low-quality papers. RESULTS: Thirty pre-vaccination studies reported frequent rotavirus strains among children below 5 years of age. G1P[8] has been identified as the most dominant type prior to vaccination in Eastern Mediterranean Region (EMR) countries. Five post-vaccination studies conducted in 3 countries (Saudi Arabia, Morocco, and Yemen) illustrated that G1P[8] is the most prevalent strain in Saudi Arabia, and the incidence of G2P[4] has increased from 21.6% to 33.3%. In Yemen, G1P[4] is the most prevalent strain (87.5%), followed by G9P[8] (57%) and G1P[8] (18.5%). Furthermore, in Yemen, G9P[8] were the most prevalent strains accounting to 57% and 14% in G9P[4], post vaccination. Finally, in Morocco, G1P[8] was not reported 3 years post vaccination; however, incidence of G9P[8] was reported at 67% and G2P[4] at 33%. CONCLUSIONS: Rotavirus circulating strain prevalence in EMR countries has changed post vaccination, and G9P[8], G2P[4], and G9P[4] have become more dominant. Proportion of rotavirus strains in these countries after vaccination has significantly reduced. There is an increase in circulating strain G2P[4] in the post-vaccination period, which needs further monitoring.

Evaluation of rapid PfHRP-2/pLDH-based tests in diagnosing microscopy-confirmed falciparum malaria in Hodeidah governorate, Yemen.

Along with the determination of malaria infection rate among suspected patients attending hospitals in Hodeidah governorate, the present study evaluated the accuracy of Plasmodium falciparum histidine-rich protein-2 (PfHRP-2)/parasite-specific lactate dehydrogenase (pLDH)-based rapid diagnostic test (RDT) for the diagnosis of microscopy-confirmed falciparum malaria. An overall malaria infection rate of 19.3% (57/295) among suspected patients attending hospitals was microscopically confirmed. The sensitivity of thin blood films for the detection of malaria parasites was 79.0% compared to thick films and was greatly affected by the parasite density, being 65.0% or less at parasite densities of ≤1000 parasites/µl of blood. Compared to light microscopy, the present study revealed sensitivity levels of 100.0% (95% CI: 92.0-100.0) vs. 94.7% (95% CI: 84.2-98.6), specificity levels of 97.3% (95% CI: 89.8-99.5) vs. 100.0% (95% CI: 93.9-100.0), positive predictive values of 89.9% (95% CI: 88.3-99.0) vs. 100.0 (95% CI: 91.6-100.0) and negative predictive values of 100.0% (95% CI: 93.9-100.0) vs. 98.7% (95% CI: 89.3-98.7) for the PfHRP-2 and pLDH components of SD BIOLINE® RDT, respectively, for falciparum malaria diagnosis. Therefore, the overall accuracy levels of the PfHRP-2 and pLDH components of the investigated RDT for the diagnosis of microscopy-confirmed falciparum malaria are 98.5% (95% CI: 94.6-99.6) and 97.7% (95% CI: 93.5-99.2), respectively.

Characterization of In vitro inhibitory effects of consensus short interference RNAs against non-structural 5B gene of hepatitis C virus 1a genotype.

PURPOSE: Chronic hepatitis C has infected approximately 170 million people worldwide. The novel direct-acting antivirals have proven their clinical efficacy to treat hepatitis C infection but still very expensive and beyond the financial range of most infected patients in low income and even resource replete nations. This study was conducted to establish an in vitro stable human hepatoma 7 (Huh-7) cell culture system with consistent expression of the non-structural 5B (NS5B) protein of hepatitis C virus (HCV) 1a genotype and to explore inhibitory effects of sequence-specific short interference RNA (siRNA) targeting NS5B in stable cell clones, and against viral replication in serum-inoculated Huh-7 cells. MATERIALS AND METHODS: In vitro stable Huh-7 cells with persistent expression of NS5B protein was produced under gentamycin (G418) selection. siRNAs inhibitory effects were determined by analysing NS5B expression at mRNA and protein level through reverse transcription-polymerase chain reaction (PCR), quantitative real-time PCR, and Western blot, respectively. Statistical significance of data (NS5B gene suppression) was performed using SPSS software (version 16.0, SPSS Inc.). RESULTS: siRNAs directed against NS5B gene significantly decreased NS5B expression at mRNA and protein levels in stable Huh-7 cells, and a vivid decrease in viral replication was also exhibited in serum-infected Huh-7 cells. CONCLUSIONS: Stable Huh-7 cells persistently expressing NS5B protein should be helpful for molecular pathogenesis of HCV infection and development of anti-HCV drug screening assays. The siRNA was effective against NS5B and could be considered as an adjuvant therapy along with other promising anti-HCV regimens.

In vitro inhibitory analysis of consensus siRNAs against NS3 gene of hepatitis C virus 1a genotype.

OBJECTIVE: To explore inhibitory effects of genome-specific, chemically synthesized siRNAs (small interference RNA) against NS3 gene of hepatitis C virus (HCV) 1a genotype in stable Huh-7 (human hepatoma) cells as well as against viral replication in serum-inoculated Huh-7 cells. METHODS: Stable Huh-7 cells persistently expressing NS3 gene were produced under antibiotic gentamycin (G418) selection. The cell clones resistant to 1000 µg antibiotic concentration (G418) were picked as stable cell clones. The NS3 gene expression in stable cell clone was confirmed by RT-PCR and Western blotting. siRNA cell cytotoxicity was determined by MTT cell proliferation assay. Stable cell lines were transfected with sequence specific siRNAs and their inhibitory effects were determined by RT-PCR, real-time PCR and Western blotting. The viral replication inhibition by siRNAs in serum inoculated Huh-7 cells was determined by real-time PCR. RESULTS: RT-PCR and Western blot analysis confirmed NS3 gene and protein expression in stable cell lines on day 10, 20 and 30 post transfection. MTT cell proliferation assay revealed that at most concentrated dose tested (50 nmol/L), siRNA had no cytotoxic effects on Huh-7 cells and cell proliferation remained unaffected. As demonstrated by the siRNA time-dependent inhibitory analysis, siRNA NS3-is44 showed maximum inhibition of NS3 gene in stable Huh-7 cell clones at 24 (80%, P = 0.013) and 48 h (75%, P = 0.002) post transfection. The impact of siRNAs on virus replication in serum inoculated Huh-7 cells also demonstrated significant decrease in viral copy number, where siRNA NS3-is44 exhibited 70% (P < 0.05) viral RNA reduction as compared to NS3-is33, which showed a 64% (P < 0.05) decrease in viral copy number. siRNA synergism (NS3-is33 + NS3-is44) decreased viral load by 84% (P < 0.05) as compared to individual inhibition by each siRNA (i.e., 64%-70% (P < 0.05)) in serum-inoculated cells. Synthetic siRNAs mixture (NS5B-is88 + NS3-is33) targeting different region of HCV genome (NS5B and NS3) also decreased HCV viral load by 85% (P < 0.05) as compared to siRNA inhibitory effects alone (70% and 64% respectively, P < 0.05). CONCLUSIONS: siRNAs directed against NS3 gene significantly decreased mRNA and protein expression in stable cell clones. Viral replication was also vividly decreased in serum infected Huh-7 cells. Stable Huh-7 cells expressing NS3 gene is helpful to develop anti-hepatitis C drug screening assays. siRNA therapeutic potential along with other anti-HCV agents can be considered against hepatitis C.

The incidence rate of thyroid cancer among women in Saudi Arabia: an observational descriptive epidemiological analysis of data from Saudi Cancer Registry 2001-2008.

This study provides a descriptive epidemiological data of thyroid cancer cases diagnosed from 2001 to 2008 among Saudi women, including the frequency and percentage of cases, the crude incidence rate (CIR) and the age-standardised incidence rate (ASIR) stratified by the region and year of diagnosis. This is a retrospective descriptive epidemiological analysis of all Saudi thyroid cancer cases recorded in the Saudi Cancer Registry (SCR) between January 2001 and December 2008. The statistical analyses were applied using descriptive statistics with the Statistical Package for the Social Sciences version 20.0. A total of 2,930 cases were recorded in the SCR between January 2001 and December 2008. The region of Riyadh in Saudi Arabia had the highest overall ASIR at 9.43 per 100,000 women, followed by Tabuk at 7.11 and eastern region at 6.5, while Jazan and Jouf had the lowest average ASIRs at 1.97 and at 2.72, respectively. The region of Qassim recorded the greatest changes of ASIR at 5.5 per 100,000 women from 2001 to 2008. There was a slight increase in the CIRs and ASIRs for thyroid cancer in Saudi Arabia between 2001 and 2008. Riyadh, Tabuk and eastern region were the highest overall ASIR in Saudi Arabia. While, Jazan and Hail had the lowest rates. Finally, the region of Qassim had the highest changes in CIR and ASIR from 2001 to 2008. Further analytical studies are needed to determine the potential risk factors of thyroid cancer disease among Saudi women.

The pattern of Middle East respiratory syndrome coronavirus in Saudi Arabia: a descriptive epidemiological analysis of data from the Saudi Ministry of Health.

PURPOSE: This study describes the epidemiology of Middle East respiratory syndrome coronavirus (MERS-CoV) in Saudi Arabia. PATIENTS AND METHODS: Epidemiological analysis was performed on data from all MERS-CoV cases recorded by the Saudi Ministry of Health between June 6, 2013 and May 14, 2014. The frequency of cases and deaths was calculated and adjusted by month, sex, age group, and region. The average monthly temperature and humidity of infected regions throughout the year was also calculated. RESULTS: A total of 425 cases were recorded over the study period. The highest number of cases and deaths occurred between April and May 2014. Disease occurrence among men (260 cases [62%]) was higher than in women (162 cases [38%]), and the case fatality rate was higher for men (52%) than for women (23%). In addition, those in the 45-59 years and ≥60 years age groups were most likely to be infected, and the case fatality rate for these people was higher than for other groups. The highest number of cases and deaths were reported in Riyadh (169 cases; 43 deaths), followed by Jeddah (156 cases; 36 deaths) and the Eastern Region (24 cases; 22 deaths). The highest case fatality rate was in the Eastern Region (92%), followed by Medinah (36%) and Najran (33%). MERS-CoV infection actively causes disease in environments with low relative humidity (<20%) and high temperature (15°C-35°C). CONCLUSION: MERS-CoV is considered an epidemic in Saudi Arabia. The frequency of cases and deaths is higher among men than women, and those above 45 years of age are most affected. Low relative humidity and high temperature can enhance the spread of this disease in the entire population. Further analytical studies are required to determine the source and mode of infection in Saudi Arabia.