Genomic analysis of SARS-CoV-2 omicron sublineage BA.5.2.1 in Erbil/Iraq.

Due to several mutations in its genomic sequence, particularly in the spike protein region, the recently-discovered SARS-CoV-2 variant B.5.2.1 has alarmed health policy authorities worldwide. The World Health Organization (WHO) has labelled it "Omicron" and classified it as a worldwide variant of concern (VOC). Following the appearance of Omicron in Iraq, new cases were also detected and analyzed in Kurdistan regions. Two hundred patients were recruited in this study from Erbil/Iraq. The RNA genome samples were extracted,  the qRT-PCR performed, and 10 samples were sequenced. The sample sequence was published (EPI ISL 15921492) in the GISAID international gene bank for COVID-19. When compared to the BA.1 Omicron sublineage, 17 new mutations and five deletions in the  Omicron subvariant BA.5.2.1 sequence were detected. The spike region includes eight of these variations and one deletion. Overall, 30 substitutions were shared between those previously seen in the BA.1 sublineage and the newly-detected BA.5.2.1 Omicron subvariant. We detected eight new substitutions in our BA.5.2.1 subvariants (T112I, A27S, V213G, T376F, D405N, R408S, L452R, F486V), which were not mentioned previously, should be cause for concern and may be related to immune escape or viral oligomerization. Omicron might be more immune-escape-capable than the current VOCs/VOIs. However, the predicted mutational research shows no conclusive evidence that the Omicron variant may be more virulent or fatal than other variations, including Delta. The greater capacity for immunological evasion may cause the current increase in Omicron cases in Erbil/Iraq.

IL1ß, IL-6, and TNF-α cytokines cooperate to modulate a complicated medical condition among COVID-19 patients: case-control study.

The cytokine storm is a complication related to SARS-CoV-2 infection worldwide. This study aimed to address the level of three cytokines which were interleukin-1-beta (IL-1ß), interleukin-6 (IL-6), and tumour necrosis factor-alpha (TNF-α), with different blood parameters to the formation of cytokine storm or any complication among COVID-19 patients. Method and material: A total of 104 serum samples were included for this purpose, and they were divided into three categories the healthy control group (n=30), mild COVID-19 patients (n=23), and severe cases of COVID-19 patients (n=51). The cytokine concentration was measured by enzyme-linked immunosorbent assays (ELISA). Serum ferritin, C-reactive protein (CRP) levels, and erythrocyte sedimentation rate were also evaluated and compared with the concentration of the proinflammatory cytokines. Result: The data analysis showed a significant relationship between the serum IL-6 level with serum ferritin and CRP and the progression to the severity of SARS-CoV-2 infection. The IL-6 level was increased in mild COVID-19 patients and was significantly elevated in severe COVID-19 patients. Patients in the severe group had significantly higher serum ferritin, CRP, and erythrocyte sedimentation rate levels than those in the mild and healthy groups. The IL-1ß and TNF-α were not significantly different in the groups compared with the healthy control group. Conclusion: This study revealed that the proinflammatory cytokines and biochemical laboratory tests are promising biomarkers for detecting the severity of COVID-19 cases.

SARS-CoV-2 Omicron Variant Genomic and Phylogenetic Analysis in Iraqi Kurdistan Region.

Omicron variants have been classified as Variants of Concern (VOC) by the World Health Organization (WHO) ever since they first emerged as a result of a significant mutation in this variant, which showed to have an impact on transmissibility and virulence of the virus, as evidenced by the ongoing modifications in the SARS-CoV-2 virus. As a global pandemic, the Omicron variant also spread among the Kurdish population. This study aimed to analyze different strains from different cities of the Kurdistan region of Iraq to show the risk of infection and the impact of the various mutations on immune responses and vaccination. A total of 175 nasopharyngeal/oropharyngeal specimens were collected at West Erbil Emergency Hospital and confirmed for SARS-CoV-2 infection by RT-PCR. The genomes of the samples were sequenced using the Illumina COVID-Seq Method. The genome analysis was established based on previously published data in the GISAID database and compared to previously detected mutations in the Omicron variants, and that they belong to the BA.1 lineage and include most variations determined in other studies related to transmissibility, high infectivity and immune escape. Most of the mutations were found in the RBD (receptor binding domain), the region related to the escape from humoral immunity. Remarkably, these point mutations (G339D, S371L, S373P, S375F, T547K, D614G, H655Y, N679K and N969K) were also determined in this study, which were unique, and their impact should be addressed more. Overall, the Omicron variants were more contagious than other variants. However, the mortality rate was low, and most infectious cases were asymptomatic. The next step should address the potential of Omicron variants to develop the next-generation COVID-19 vaccine.

Human cytomegalovirus UL24 and UL43 products participate in SAMHD1 subcellular localization.

This report has analyzed the potential role of Human Cytomegalovirus (HCMV) UL24 and UL43 products in modulating the subcellular location of a host restriction factor, SAMHD1, in cells of human fibroblast origin. Recent studies have reported that the regulation of SAMHD1 is mediated by the HCMV UL97 product inside the nucleus, and by the CDK pathway when it is located in the cytoplasm of the infected cells but the viral gene products that may involve in cytosolic relocalization remain unknown yet. In the present report, we demonstrate that the HCMV UL24 product interacts with the SAMHD1 protein during infection based on mass spectrometry (MS) data and immunoprecipitation assay. The expression or depletion of the viral UL24 gene product did not affect the subcellular localization of SAMHD1 but when it coexpressed with the viral UL43 gene product, another member of the HCMV US22 family, induced the SAMHD1 cytosolic relocalization. Interestingly, the double deletion of viral UL24 and UL43 gene products impaired the cytosolic translocation and the SAMHD1 was accumulated in the nucleus of the infected cells, especially at the late stage post-infection. Our results provide evidence that the viral UL24 and UL43 gene products play a role in the SAMHD1 subcellular localization during HCMV infection. Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-022-00799-3.

Human Cytomegalovirus UL24 and UL43 Cooperate to Modulate the Expression of Immunoregulatory UL16 Binding Protein 1.

The human cytomegalovirus (HCMV) UL24 and UL43 are tegument proteins that have recently been shown to interact with each other in a yeast two-hybrid system. By their overexpression in MRC5 cells, we demonstrate that these viral proteins interact with several important host proteins, especially Dicer and trans-activation response RNA binding protein. As these hots proteins are involved in regulating the production of cellular micro-RNAs, the cytomegalovirus (CMV) proteins could interfere with their actions to favor viral replication directly or through an immune escape mechanism. Double knockout of UL24 and UL43 does not show a remarkable effect on CMV entry or replication, but it significantly downregulates the expression of CMV-encoded miR-UL59, which is thought to regulate the expression of a downstream target UL16 binding protein 1 (ULBP1). Interestingly, the double knockout increases the expression of the ULBP1 recognized by the NKG2D activating receptor of natural killer cells. This study investigates the potential role of several proteins encoded by HCMV in regulating the host cellular environment to favor escape from immunity, and it also provides some basis for the future development of RNA-targeted small molecules to control HCMV infection.

Dissemination and Genetic Relatedness of Multidrug-Resistant and Extensively Drug-Resistant Acinetobacter baumannii Isolates from a Burn Hospital in Iraq.

Acinetobacter baumannii is an aggressive opportunistic bacterial pathogen that causes severe nosocomial infections, especially among burn patients. An increasing number of hospitals-acquired infections have been reported all over the world. However, little attention has been paid to the relatedness between A. baumannii isolates from different hospital environments and patients. In this study, 27 isolates were collected from the Burn and Plastic Surgery Hospital of Al Sulaymaniyah City, Iraq, from January through December 2019 (11 from patients and 16 from the wards environment), identified to species level as A. baumannii using Vitek 2 system and molecular detection of 16S rRNA gene, and then confirmed by targeting the bla OXA-51 gene. Moreover, the isolates were characterized by means of automated antimicrobial susceptibility assay, antimicrobial-resistant patterns, a phenotypic method using a combined disk test, and molecular methods for the detection of class A and C ß-lactamase genes, and finally, the genetic relatedness was classified. Antimicrobial susceptibility testing showed that 63% (17/27) of the retrieved A. baumannii isolates were extensively drug-resistant to 8/9 antimicrobial classes. Furthermore, 37% (10/27) of the isolates were classified as multidrug-resistant; 8 isolates exhibited similar resistant patterns and the other two isolates showed 2 different patterns, while resistance was greater in isolates from patients than from the ward environment. Combined disk test showed that two isolates contained extended-spectrum ß-lactamase. All isolates carried bla TEM-1, and two copies of the bla CTX-1 gene were indicated in one isolate, while bla SHV was absent in all isolates. Twenty-four isolates carried the bla AmpC gene; among them, 3 isolates harbored the insertion sequence ISAba-1 upstream to the gene. Using Enterobacterial Repetitive Intergenic Consensus PCR, the isolates were clustered into 6 distinct types; among them, two clusters, each of four strains, were classified to contain isolates from both patients and environments. The clusters of similar genotypes were found in inpatients as well as the environments of different wards during time periods, suggesting transmission within the hospital. Identification of possible infection sources and controlling the transmission of these aggressive resistance strains should be strictly conducted.

MicroRNAs in the development of potential therapeutic targets against COVID-19: A narrative review.

BACKGROUND: As the therapeutic regimens against the COVID-19 remain scarce, the microRNAs (miRNAs) can be exploited to generate efficient therapeutic targets. The miRNAs have been found to play pivotal roles in the several regulatory functions influencing the prognosis of viral infection. The miRNAs have a prospective role in the up and down regulation of the ACE2 receptors. This review examines the clinical applications, as well as the possible threats associated with the use of miRNAs to combat the deleterious consequences of SARS-CoV-2 infection. METHODOLOGY: This article was compiled to evaluate how the miRNAs are involved in the SARS-CoV-2 pathogenesis and infection, and their potential functions which could help in the development of therapeutic targets against the COVID-19. The sources of the collected information include the several journals, databases and scientific search engines such as the Google scholar, Pubmed, Science direct, official website of WHO, among the other sites. The investigations on the online platform were conducted using the keywords miRNA biogenesis, miRNA and ACE2 interaction, therapeutic role of miRNAs against SARS-CoV-2 and miRNA therapy side effects. RESULTS: This review has highlighted that the miRNAs can be exploited to generate potential therapeutic targets against the COVID-19. Changes in the miRNA levels following viral replication are an essential component of the host response to infection. The collection and modification of miRNA modulates may help to minimize the deleterious consequences of SARS-CoV-2 infection, such as by controlling or inhibiting the generation of cytokines and chemokines. The degradation of viral RNA by the cellular miRNAs, along with the reduced expression of ACE2 receptors, can substantially reduce the viral load. Specific miRNAs have been found to have an antiviral influence, allowing the immune system to combat the infection or forcing the virus into a latency stage. CONCLUSION: This review summarizes several studies revealing the involvement of miRNAs in diverse and complex processes during the infection process of SARS-CoV-2. The miRNAs can substantially reduce the viral load by degradation of viral RNA and reduced expression of ACE2 receptors, besides mitigating the deleterious consequences of the exaggerated secretion of cytokines. Extensive investigations need to be done by the scientific community to utilize the miRNA based strategies for the development of effective therapeutic targets against the COVID-19.

SARS-CoV-2 vaccine breakthrough reinfection in a health-care worker of Iraq: A case report.

The COVID-19 pandemic has severely affected the entire globe since the first isolation of SARS-CoV-2 from patients with severe respiratory illness in Wuhan, China. Although the global vaccination drive is in full swing, many cases of reinfection have also been reported after vaccination. Currently, there is a scarcity of data available on the reinfection and vaccine breakthrough infections in Iraq. In this letter, we have presented a case report on the SARS-CoV-2 vaccine breakthrough reinfection in a health-care worker after completion of the double-dose vaccination. An increased symptom severity was reported on the second infection, which was confirmed to be of Delta variant. Such vaccine breakthrough infection reports have raised important questions regarding the duration of vaccine-mediated immunity and vaccine effectiveness against all circulating variants. These have further emphasized the importance of following non-pharmaceutical interventions by fully vaccinated individuals, especially at health-care settings.

Strategies to overcome the main challenges of the use of CRISPR/Cas9 as a replacement for cancer therapy.

CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats-associated protein 9) shows the opportunity to treat a diverse array of untreated various genetic and complicated disorders. Therapeutic genome editing processes that target disease-causing genes or mutant genes have been greatly accelerated in recent years as a consequence of improvements in sequence-specific nuclease technology. However, the therapeutic promise of genome editing has yet to be explored entirely, many challenges persist that increase the risk of further mutations. Here, we highlighted the main challenges facing CRISPR/Cas9-based treatments and proposed strategies to overcome these limitations, for further enhancing this revolutionary novel therapeutics to improve long-term treatment outcome human health.

Olfactory dysfunction as a post-infectious symptom of SARS-CoV-2 infection.

The unexpected onset smell and taste disability was being recognized as a COVID-19 related symptom. Loss of smell might occur alone or be followed by other COVID-19 symptoms, such as a dry cough, fever, headache, and shortness of breath. Other virus infections have been linked to anosmia (parainfluenza, rhinovirus, SARS, and others), affecting up to 20% of the adult population, which is much less common than SARS-CoV-2 infection. A hypothesis about the pathophysiology of post-infectious olfactory loss is that viruses could make an inflammatory response of the nasal mucosa or directly damage the olfactory neuroepithelium. However, in patients with COVID-19, loss of smell may occur without other rhino logic symptoms or suggestive nasal inflammation. According to evidence, anosmia-related SARS-CoV-2 could be a new viral syndrome unique to COVID-19. Furthermore, through experimental intranasal inoculation in mice, SARS-CoV-2 can be inoculated into the olfactory neural circuitry. This disease has not had the required focus, most likely because it is not life-threatening in and of itself. Though patients' quality of living is significantly reduced as their olfactory ability is lost, resulting in lowering and inadequate appetite, excessive or unbalanced food consumption, as well as an overall sense of insecurity. This review aims to give a quick overview of the latest epidemiological research, pathological mechanisms for the dysfunction of smell, and taste in patients infected with SARS-CoV-2. In addition, the initial diagnosis and treatment options for dysfunction are also discussed.

Nanopore sequencing from extraction-free direct PCR of dried serum spots for portable hepatitis B virus drug-resistance typing.

BACKGROUND: Effective drug regimens for the treatment of hepatitis B virus (HBV) infections are essential to achieve the World Health Organisation commitment to eliminate viral hepatitis by 2030. Lamivudine (3TC) is widely used in countries with high levels of chronic HBV, however resistance has been shown to occur in up to 50 % of individuals receiving continuous monotherapy for 4 years. Telbivudine (LdT) is now more commonly used in place of lamivudine but is ineffective against 3TC-resistant HBV. Genotyping and identification of resistanceassociated substitutions (RAS) is not practical in many locations. OBJECTIVES: A novel assay was designed to enable HBV genotyping and characterisation of resistance mutations directly from serum samples stored on filter paper, using Sanger and MinION sequencing. STUDY DESIGN: The assay was applied to a cohort of 30 samples stored on filter paper for several years with HBV viral loads ranging from 8.2 × 108 to 635 IU/mL. A set of 6 high-titre samples were used in a proof-of-principle study using the MinION sequencer. RESULTS: The assay allowed determination of HBV genotype and elucidation of RAS down to 600 IU/mL using a 550bp amplicon. Sequencing of a 1.2 kb amplicon using a MinION sequencer gave results consistent with Sanger sequencing and allowed the identification of minor populations of variants. CONCLUSIONS: We present two approaches for reliable HBV sequencing and RAS identification using methods suitable for resource-limited environments. This is the first demonstration of extraction-free DNA sequencing direct from DSS using MinION and these workflows are adaptable to the investigation of other DNA viruses.

Elevated serum activity of MBL and ficolin-2 as biomarkers for progression to hepatocellular carcinoma in chronic HCV infection.

Hepatocellular carcinoma (HCC) is an uncommon but significant outcome of chronic hepatitis C virus (HCV) infection. A serum biomarker for predicting progression to HCC would have a major impact on patient monitoring and clinical management. We explored circulating liver-expressed lectins, ficolin-2, ficolin-3 and mannose binding lectin (MBL), as potential biomarkers for the development of HCC. The activity of these three lectins were analysed in HCV positive patients who developed HCC (n = 31) with comparable HCV-positive HCC-negative patients (n = 106) and healthy controls (n = 79). Serum binding activity of ficolin-2 and MBL were elevated compared to controls. Analysis of pre-HCC onset samples revealed that MBL levels were significantly elevated up to 3 years, and ficolin-2 was elevated up to 1 year, prior to diagnosis of HCC over controls. This preliminary study identifies MBL and ficolin-2 as potential biomarkers for the development of HCC in chronic HCV infection.

Expression of human ficolin-2 in hepatocytes confers resistance to infection by diverse hepatotropic viruses.

The liver-expressed pattern recognition receptors mannose-binding lectin (MBL), ficolin-2 and ficolin-3 contribute to the innate immune response by activating complement. Binding of soluble ficolin-2 to viral pathogens can directly neutralize virus entry. We observed that the human hepatoma cell line HuH7.5, which is routinely used for the study of hepatotropic viruses, is deficient in expression of MBL, ficolin-2 and ficolin-3. We generated a cell line that expressed and secreted ficolin-2. This cell line (HuH7.5 [FCN2]) was more resistant to infection with hepatitis C virus (HCV), ebolavirus and vesicular stomatitis virus, but surprisingly was more susceptible to infection with rabies virus. Cell-to-cell spread of HCV was also inhibited in ficolin-2 expressing cells. This illustrates that ficolin-2 expression in hepatocytes contributes to innate resistance to virus infection, but some viruses might utilize ficolin-2 to facilitate entry.