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Islam has divided the obligatory duties into Huquq ul Allah and Huquq–al-Ibad­ and complying with them guaranteed the success in this world and the hereafter. Islam not only connects rights and responsibilities with others, but also determines their priorities. Those nations where imbalance is created in discharging duties and rights get caught up in mischief and trouble as an unavoidable consequence as if human beings play the main role in the construction and destruction of the society peace. Maintaining justice as a whole is a vital part of an Islamic state; Allah (SWT) has ordered to show justice even with enemies. Justice cannot be denied in any case. In Islam the implementation of justice is as much important as the wreaking of a father cannot be taken from his son. Other religions also give importance to justice. Before Islam people had forgotten these principles but with the advent of Islam, the discrimination between rich and poor, master and slave was removed because favoring one side would create hatred and encourage class discrimination and would ultimately result in the decline of any state. Therefore, to protect the society from all such evils, Islam has laid importance on following rules of justice. In this perspective, the article describes the importance of justice, equality, reconstruction of judiciary and judicial system in an Islamic state., http://ojs.uopisl.pk/index.php/peshawarislamicus/article/view/80

The objective of the study is to measure product diversification’s impact on insurance firm’s financial performance in Pakistan. Analysis are carried out to examine how ownership structure, capitalization, group membership, firm size, diversification across business lines, industry concentration affects firm’s financial performance. Data from 2009-2019 is collected to measure the impact of diversification (entropy) on the risk- adjusted returns. Findings of the study reveal that business line diversification has strong positive effect on firm performance (for both ROA and ROE) which means that diversified firms perform better than non-diversified firms. For managers these findings are useful as they propose the need for diversification, capitalization, increase in size and group affiliation to enhance firm profitability.

Introduction The COVID-19 pandemic has had an extensive impact on the frail older population, with significant rates of COVID-related hospital admissions and deaths amongst this vulnerable group. There is little evidence comparing the prevalence and impact of frailty amongst patients hospitalised with COVID-19 in wave 1 vs wave 2 of the pandemic. Methods Prospective observational study of all consecutive patients admitted to Salford Royal NHS Foundation Trust (SRFT) between 27th February and 28th of April 2020 (wave 1), and 1st October to 10th November 2020 (wave 2) with a diagnosis of COVID-19. The primary endpoint was in-hospital mortality. Patient demographics, co-morbidities, biochemical parameters, and frailty (using the Clinical Frailty Scale, score 1–4 = not frail, score 5–9 = frail) were collected. A Cox proportional hazards model associating wave and frailty with mortality was used. A logistic regression model was used to associate patient characteristics with wave. Both models adjusted for patient characteristics. Results A total of 700 patients were included (N = 429, wave 1; N = 271, wave 2). In wave 1, 42% (N = 180) were female; median age was 72; 37% (N = 160) were non-survivors, 49% (N = 212) were frail (CFS 5–9). In wave 2, 38% (N = 104) were female; median age was 73; 30% (N = 80) were non-survivors, 39% (N = 106) were frail. There was a reduction in mortality in wave 2, aHR = 0.71 (95% CI 0.53–0.94). Frailty was associated with increased mortality, after adjustment for age, wave and other patient characteristics. Patients were more frail in wave 1, and the effect of frailty was more pronounced in wave 1 vs wave 2. Conclusion Frailty is highly prevalent amongst patients of all ages admitted to SRFT with COVID-19. Higher scores of frailty are associated with increased mortality.

Femoral neck fractures are one of the most common fractures treated by an Orthopaedic surgeon. Arthroplasty is the recommended management for intracapsular neck of femur fractures in the elderly population owing to the high risk of avascular necrosis of the femoral head following an internal fixation. Elderly patients with intracapsular fractures deemed high risk for anaesthesia (American Society of Anaesthesiology Grade more than 2) are recommended a hip hemiarthroplasty. Routine practice throughout the United Kingdom is to obtain a postoperative check radiograph for all hip hemiarthroplasty patients prior to their discharge from the hospital. This may be done for various reasons like checking the presence of any peri-prosthetic fracture, the position of the components along with the presence of any dislocation. However, it is unclear whether a radiograph is the sole identifier of such complications. Through this study, we aim to analyse whether routine recommendation of post-operative radiographs following hip hemiarthroplasty affects the clinical outcome, and whether it is effective in identifying potential complications before the patients report any signs or symptoms.

Objective The goal of the study is to find out the treatment of choice for ankle fractures involving syndesmotic injury based on level of experience of orthopaedic surgeons. Methods A survey was undertaken to analyse the management used for ankle fractures with syndesmotic injuries AO 44c in a 35-year-old patient. Surgeons attending an orthopaedic course were invited to take part in a survey sorted into groups: junior surgeons middle grades, and experienced. Pictures of an x ray were shown to the participants and treatment options were asked. Results 100 surgeons from 20 nations took part in the event. Juniors made up 39%, registrars made up 38%, and experienced doctors were 29%. Screws, were reported by 93% for syndesmosis fixation. 66% of surgeons who used screws for syndesmosis fixation favoured a single screw over two screws.3-4 cortices were virtually evenly divided in choice, with 54% preferring three and 46% preferring four cortices. Only 22% of the time did they utilise a washer with their screws. With 52% of patients, the most typical time for permitting them to weight bear was 4–6 weeks after surgery. At 1–2 months postoperatively, 34% preferred to remove the screw, and at 2-3 months postoperatively, 29% preferred to remove the screw. Conclusion Data show that the majority of junior level doctors handle their patients according to AO principles. The majority prefer one 3.5 mm screw positioned between 2 and 4 cm above the ankle joint, with three cortices being somewhat preferred. Despite the lack of data to support one procedure, the majority of people remove their screws within 1–3 months, Highlights • Various techniques have been described in literature for fixation of ankle fractures with syndesmotic injury. • To assess the management of AO 44c syndesmotic injury, a survey of participants at an international AO course was done. • The goal is to identify the treatment of choice based on surgeon’s experience to treat ankle fracture with syndesmotic injury.

Nanotechnology holds great potential in advancing wastewater treatment to improve treatment efficiency and to supplement the water supply. Nanomaterials based on metallic, carbonaceous, polymeric, and magnetic nanoadsorbents are discussed in detail in this chapter, along with nanofiltration membranes, thin-film composite membranes, fabricated membranes, nanocomposite membranes, polymeric nanocomposite membranes, and enzyme incorporated nanotechnology-based processes. Some conventional treatment processes including coagulation-flocculation for removal of organic and inorganic pollutants along with suspended solid particles are still in use for water treatment. The recent advancements in the area of coagulation-flocculation for wastewater treatment are critically reviewed and evaluated. This chapter also emphasizes destabilization of colloids, removal of inorganic and organic matter (particulate and/or dissolved), removal of metals and anions, and removal of microorganisms using coagulation-flocculation. Various chemical or natural coagulants, flocculants and their derivatives and the use of low-cost biopolymeric materials such as algin, guar gum pectin, starch, chitin, dextran, and pullulan derivatives are also described in detail.

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Changing times require industries to adopt ever-changing technologies in order to stay afloat and compete at the global stage. The logistics industry is experiencing a global boom due to progress in the e-commerce sector and ever-increasing consumer needs of the modern world. Oman’s long-term vision includes logistic sector as one the main contributors to the country’s GDP. Sea Ports such as Sohar and Al Duqm are another indication of the Sultanate’s ambitions to diversify the economy for the post-oil era. The logistics sector in Oman needs to adopt technology that not only helps it with improving its operations but helps reshape the industry standards in Oman. Robot adoption is the latest trend in the global logistics industry and Oman’s leading logistics companies need to adopt it too. The combination of logistic robots and right artificial intelligent programming can do wonders for a company. Artificial intelligence complements robotics to achieve their tasks more efficiently and helps with better robot maintenance. Oman’s logistics sector can benefit greatly from the introduction of logistic robots on both practical fronts as well as the marketing front. A research was conducted based on one of Oman’s leading and most prominent logistics company to identify the level of robotics and artificial intelligence in use. The descriptive approach of research was applied to attain the required data through both quantitative and qualitative means. Interviews and surveys were conducted in order to justify the requirement of the proposed research. The aim of the study was to build a case for implementation of robotics in conjunction with artificial intelligence in Oman’s logistics sector in order for Oman’s logistics sector to fulfil its potential and compete at global stage.

Background In orthopedic surgery, bleeding is an inevitable side effect. The study's aim was to provide estimated blood loss values in various orthopedic procedures and take a step towards developing statistically reliable formulae. This can provide blood loss values in orthopedic surgery, which will be a very good tool for operative planning. Materials and methods We reviewed case notes of 282 patients in a UK based trauma center from December 2020 to March 2021,who had undergone a various orthopedic procedures. The results were analyzed using SPSS version 25. Results Most common fracture was neck of femur (37.5%)followed by intertrochanteric fractures(27.6%). Paired t-test was used, and there is good evidence (t281 = 14.957, p = 0.000) that intraoperative transfusions increased HB levels in patients (t281 = 14.957, p = 0.000) by an average of 1.331 points, with a 95% confidence interval of 1.156–1.506. As a result, the variation between the Pre-op and Post-op HB levels is statistically important but minimal. We can see that the mean blood loss is statistically different in different age groups (0.03) of patients and by the existence of co-morbids using analysis of variance (0.04). The average number of days spent in the hospital varies by surgical type (0.01) performed on patients. Conclusion Orthopedic surgery can be associated with high levels of blood loss. There is a significant relation between fracture form and age groups, change of wound dressing (COD), use of a tourniquet, and drain insertion, no connection was noted between gender and fracture types., Highlights • Bleeding is an unavoidable side effect of most surgical operations, including orthopaedic surgery. • There is a scarcity of literature that can provide us an estimate of how much blood will be lost during various orthopaedic operations. • Goal of the study was to offer estimated blood loss estimates in various orthopaedic operations. • There is a significant relationship between fracture type and age groups, change of wound dressing (COD), use of a tourniquet.

Background Consent is a process of communication and the consent form is an important legal document of the evidence of discussion between doctor and patients. We observed frequent use of abbreviations in the consent forms in our department that can result in misunderstanding and miscommunication when consenting patients for orthopaedic procedures. Methods We completed an audit cycle starting by reviewing a total of 350 consent forms retrospectively in level one trauma centres in October–November of 2019 for different orthopaedic trauma procedures. The standards for the project were guidelines published by the general medical council (GMC), The royal college of surgeons (RCS) Glasgow, and the British orthopaedic association (BOA).The results were presented at our mortality and morbidity meeting. Written Feedback was obtained from the attending members on how a change can be implemented to increase ccompliance in filling consent forms. A generic email was sent to all medical professionals to avoid the use of abbreviations on the document and encourage colleagues to point out errors if they spot them. The use of full medical terms and to avoid abbreviations in consent form was well advertised, The re-audit was performed for the period of January & February 2020 that included 400 consent forms. The results were analysed and compared with our original audit results. Results The use of abbreviations declined from 54% in first audit to 22% in the re-audit. DVT and PE were the most common abbreviations. Conclusion This audit cycle has shown the importance of education and reminders to the health professionals in achieving better adherence to the guidelines and improves patient care., Highlights • Consent is as important as any basic principle on which surgical practice is carried out. • There are many different guidelines in place to prevent using abbreviations. • Use of simple language is important for good understanding of the procedure especially in consenting. • Our study looked at different abbreviations used and how they can be prevented.

Vatreptacog alfa (VA), a recombinant activated human factor VII (rFVIIa) variant with 3 amino acid substitutions, was developed to provide increased procoagulant activity in hemophilia patients with inhibitors to factor VIII or factor IX. In phase 3 clinical trials, changes introduced during the bioengineering of VA resulted in the development of undesired anti-drug antibodies in some patients, leading to the termination of a potentially promising therapeutic protein product. Here, we use preclinical biomarkers associated with clinical immunogenicity to validate our deimmunization strategy applied to this bioengineered rFVIIa analog. The reengineered rFVIIa analog variants retained increased intrinsic thrombin generation activity but did not elicit T-cell responses in peripheral blood mononuclear cells isolated from 50 HLA typed subjects representing the human population. Our algorithm, rational immunogenicity determination, offers a broadly applicable deimmunizing strategy for bioengineered proteins.

Background & Aims Induction of cross-reactive antibodies targeting conserved epitopes of the envelope proteins E1E2 is a key requirement for an hepatitis C virus vaccine. Conserved epitopes like the viral CD81-binding site are targeted by rare broadly neutralizing antibodies. However, these viral segments are occluded by variable regions and glycans. We aimed to identify antigens exposing conserved epitopes and to characterize their immunogenicity. Methods We created hepatitis C virus variants with mutated glycosylation sites and/or hypervariable region 1 (HVR1). Exposure of the CD81 binding site and conserved epitopes was quantified by soluble CD81 and antibody interaction and neutralization assays. E2 or E1-E2 heterodimers with mutations causing epitope exposure were used to immunize mice. Vaccine-induced antibodies were examined and compared with patient-derived antibodies. Results Mutant viruses bound soluble CD81 and antibodies targeting the CD81 binding site with enhanced efficacy. Mice immunized with E2 or E1E2 heterodimers incorporating these modifications mounted strong, cross-binding, and non-interfering antibodies. E2-induced antibodies neutralized the autologous virus but they were not cross-neutralizing. Conclusions Viruses lacking the HVR1 and selected glycosylation sites expose the CD81 binding site and cross-neutralization antibody epitopes. Recombinant E2 proteins carrying these modifications induce strong cross-binding but not cross-neutralizing antibodies. Lay summary Conserved viral epitopes can be made considerably more accessible for binding of potently neutralizing antibodies by deletion of hypervariable region 1 and selected glycosylation sites. Recombinant E2 proteins carrying these mutations are unable to elicit cross-neutralizing antibodies suggesting that exposure of conserved epitopes is not sufficient to focus antibody responses on production of cross-neutralizing antibodies.

Posttranslational modifications (PTMs) are often required for proper folding and physiological function of proteins, including the envelope glycoproteins 1 and 2 (E1 and E2) of hepatitis C virus (HCV). Commonly used expression systems such as bacteria, yeast, and baculovirus produce soluble HCV E1 and E2 at very low yields, as the cellular environment and molecular machinery necessary for PTMs may be suboptimal or missing. Here, we describe an expression system for HCV E2 ectodomain (eE2) with 11 N-linked glycans and eight disulfide bonds, which combines lentivirus transduction of mammalian cells and a continuous growth, adherent cell bioreactor. It is environmentally friendly, as well as cost- and time-efficient compared to other methods of recombinant protein expression in mammalian systems with final yields of eE2 approaching 60 mg/L of cell culture supernatant. eE2 produced by this system is amenable to a variety of biophysical studies, including structural determination by X-ray crystallography. Considering the ease of use and flexibility, this method can be applied to express an array of difficult target proteins in a variety of mammalian cell lines.

Hepatitis C virus (HCV) enters human hepatocytes using four essential entry factors, one of which is human CD81 (hCD81). The tetraspanin hCD81 contains a large extracellular loop (LEL), which interacts with the E2 glycoprotein of HCV. The role of the non-LEL regions of hCD81 (intracellular tails, four transmembrane domains, small extracellular loop and intracellular loop) is poorly understood. Here, we studied the contribution of these domains to HCV susceptibility of hepatoma cells by generating chimeras of related tetraspanins with the hCD81 LEL. Our results show that non-LEL regions in addition to the LEL determine susceptibility of cells to HCV. While closely related tetraspanins (X. tropicalis CD81 and D. rerio CD81) functionally complement hCD81 non-LEL regions, distantly related tetraspanins (C. elegans TSP9 amd D. melanogaster TSP96F) do not and tetraspanins with intermediate homology (hCD9) show an intermediate phenotype. Tetraspanin homology and susceptibility to HCV correlate positively. For some chimeras, infectivity correlates with surface expression. In contrast, the hCD9 chimera is fully surface expressed, binds HCV E2 glycoprotein but is impaired in HCV receptor function. We demonstrate that a cholesterol-coordinating glutamate residue in CD81, which hCD9 lacks, promotes HCV infection. This work highlights the hCD81 non-LEL regions as additional HCV susceptibility-determining factors.

RIG-I (Retinoic Acid Inducible Gene-I) is a cytosolic innate immune receptor that detects atypical features in viral RNAs as foreign to initiate a Type I interferon signaling response. RIG-I is present in an autoinhibited state in the cytoplasm and activated by blunt-ended double-stranded (ds)RNAs carrying a 5′ triphosphate (ppp) moiety. These features found in many pathogenic RNAs are absent in cellular RNAs due to post-transcriptional modifications of RNA ends. Although RIG-I is structurally well characterized, the mechanistic basis for RIG-I's remarkable ability to discriminate between cellular and pathogenic RNAs is not completely understood. We show that RIG-I's selectivity for blunt-ended 5′-ppp dsRNAs is ≈3000 times higher than non-blunt ended dsRNAs commonly found in cellular RNAs. Discrimination occurs at multiple stages and signaling RNAs have high affinity and ATPase turnover rate and thus a high katpase/Kd. We show that RIG-I uses its autoinhibitory CARD2-Hel2i (second CARD-helicase insertion domain) interface as a barrier to select against non-blunt ended dsRNAs. Accordingly, deletion of CARDs or point mutations in the CARD2-Hel2i interface decreases the selectivity from ≈3000 to 150 and 750, respectively. We propose that the CARD2-Hel2i interface is a ‘gate’ that prevents cellular RNAs from generating productive complexes that can signal.

SummaryHepatitis C virus (HCV) enters human hepatocytes through a multistep mechanism involving, among other host proteins, the virus receptor CD81. How CD81 governs HCV entry is poorly characterized, and CD81 protein interactions after virus binding remain elusive. We have developed a quantitative proteomics protocol to identify HCV-triggered CD81 interactions and found 26 dynamic binding partners. At least six of these proteins promote HCV infection, as indicated by RNAi. We further characterized serum response factor binding protein 1 (SRFBP1), which is recruited to CD81 during HCV uptake and supports HCV infection in hepatoma cells and primary human hepatocytes. SRFBP1 facilitates host cell penetration by all seven HCV genotypes, but not of vesicular stomatitis virus and human coronavirus. Thus, SRFBP1 is an HCV-specific, pan-genotypic host entry factor. These results demonstrate the use of quantitative proteomics to elucidate pathogen entry and underscore the importance of host protein-protein interactions during HCV invasion.

Non-alcoholic fatty liver disease prevalence has not been well established. The aims of this study was to define prospectively non-alcoholic steatohepatitis (NASH) and non-alcoholic fatty liver disease (NAFLD) prevalence in hospitalized and ambulatory patients 20-65 years old during June 2013 to June 2014 were selected from Combined Military Hospital Peshawar Cantonment area. A base line questionnaire and right upper quadrant ultrasound was completed by all patients. On identifications of fatty liver among the selected cases further lab test data and liver biopsy reports were obtained. Mean BMI of female was 29.9 + 5.65 while prevalence of hypertension and diabetes was 49.8% and 16.6% respectively. Among all patients 62% were Punjabies, 23% were Pathans while 12% were Sindhies. Overall NAFLD prevalence was 47% while NASH was confirmed in 20 patients (12.3% of total and 30%of ultrasound positive patients). Pathans had the highest prevalence of NAFLD (58.5%) as compared to Punjabies (44.5%) and Sindhies (35.3%). Pathans also had a higher prevalence of NASH compared with Punjabies (19.5% VS 10%: P= 0.03). In general, NAFLD patients were more prevalent among male (59%), Diabetic (P0.00005), hypertensive (P0.00005) and older (P =0.005). They consumed more fast food (P=.049) had a higher BMI (P0.0005) and had little or no exercise as compared to their normal or non NAFLD counter parts (P=0.02). NAFLD was found in 75% and NASH in 22.5% among the 26 diabetic patients. ALT, AST, BMI, insulin, quantitative insulin sensitivity checks index and cytokeratin - 18 correlated with NASH. It was concluded that NAFLD and NASH prevalence is higher than estimated previously, Pathans and Patients with diabetes are at high risk.

The hepatitis C virus (HCV) glycoprotein E2 is the major target of neutralizing antibodies and is therefore highly relevant for vaccine design. Its structure features a central immunoglobulin (Ig)-like β-sandwich that contributes to the binding site for the cellular receptor CD81. We show that a synthetic peptide corresponding to a β-strand of this Ig-like domain forms an α-helix in complex with the anti-E2 antibody DAO5, demonstrating an inside-out flip of hydrophobic residues and a secondary structure change in the composite CD81 binding site. A detailed interaction analysis of DAO5 and cross-competing neutralizing antibodies with soluble E2 revealed that the Ig-like domain is trapped by different antibodies in at least two distinct conformations. DAO5 specifically captures retrovirus particles bearing HCV glycoproteins (HCVpp) and infectious cell culture-derived HCV particles (HCVcc). Infection of cells by DAO5-captured HCVpp can be blocked by a cross-competing neutralizing antibody, indicating that a single virus particle simultaneously displays E2 molecules in more than one conformation on its surface. Such conformational plasticity of the HCV E2 receptor binding site has important implications for immunogen design., IMPORTANCE Recent advances in the treatment of hepatitis C virus (HCV) infection with direct-acting antiviral drugs have enabled the control of this major human pathogen. However, due to their high costs and limited accessibility in combination with the lack of awareness of the mostly asymptomatic infection, there is an unchanged urgent need for an effective vaccine. The viral glycoprotein E2 contains regions that are crucial for virus entry into the host cell, and antibodies that bind to these regions can neutralize infection. One of the major targets of neutralizing antibodies is the central immunoglobulin (Ig)-like domain within E2. We show here that this Ig-like domain is conformationally flexible at the surface of infectious HCV particles and pseudoparticles. Our study provides novel insights into the interactions of HCV E2 with the humoral immune system that should aid future vaccine development.

The major group human rhinovirus type 8 can enter cells via heparan sulphate. When internalized into ICAM-1 negative rhabdomyosarcoma (RD) cells, HRV8 accumulated in the cells but caused CPE only after 3days when used at high MOI. Adaptation by three blind passages alternating between RD and HeLa cells resulted in variant HRV8v with decreased stability at acidic pH allowing for productive infection in the absence of ICAM-1. HRV8v produced CPE at 10 times lower MOI within 1day. Confocal fluorescence microscopy colocalization and the use of pharmacological and dominant negative inhibitors revealed that viral uptake is clathrin, caveolin, and flotillin independent. However, it is blocked by dynasore, amiloride, and EIPA. Furthermore, HRV8v induced FITC-dextran uptake and colocalized with this fluid phase marker. Except for the complete inhibition by dynasore, the entry pathway of HRV8v via HS is similar to that of HRV14 in RD cells that overexpress ICAM-1.

Stress, work-family conflicts and flexible scheduling are three of the most important elements in organizational studies. The focus of current study is to understand the effect of Stress,work family conflicts and flexible scheduling on employee’s performance and also to understand whether flexible scheduling helps in reducing stress and work-family conflicts or not. The back bone of this study is the secondary data comprised of comprehensive literature review. A survey has also been conducted to strengthen the idea comprising of a sample of 70 employees from different organizations. 53 of them responded and the respond rate was 75%. Descriptive statistics is used to analyze the data. Results show that stress and work family conflict negatively affect the employee performance and flexible scheduling has a positive effect on employee performance. Primary study as well as literature review showed that flexible scheduling also helps in reducing stress and work-family conflicts. However, results are strongly based on the literature review i.e. secondary data.

RNAs with 5′-triphosphate (ppp) are detected in the cytoplasm principally by the innate immune receptor Retinoic Acid Inducible Gene-I (RIG-I), whose activation triggers a Type I IFN response. It is thought that self RNAs like mRNAs are not recognized by RIG-I because 5′ppp is capped by the addition of a 7-methyl guanosine (m7G) (Cap-0) and a 2′-O-methyl (2′-OMe) group to the 5′-end nucleotide ribose (Cap-1). Here we provide structural and mechanistic basis for exact roles of capping and 2′-O-methylation in evading RIG-I recognition. Surprisingly, Cap-0 and 5′ppp double-stranded (ds) RNAs bind to RIG-I with nearly identical Kd values and activate RIG-I’s ATPase and cellular signaling response to similar extents. On the other hand, Cap-0 and 5′ppp single-stranded RNAs did not bind RIG-I and are signaling inactive. Three crystal structures of RIG-I complexes with dsRNAs bearing 5′OH, 5′ppp, and Cap-0 show that RIG-I can accommodate the m7G cap in a cavity created through conformational changes in the helicase-motif IVa without perturbing the ppp interactions. In contrast, Cap-1 modifications abrogate RIG-I signaling through a mechanism involving the H830 residue, which we show is crucial for discriminating between Cap-0 and Cap-1 RNAs. Furthermore, m7G capping works synergistically with 2′-O-methylation to weaken RNA affinity by 200-fold and lower ATPase activity. Interestingly, a single H830A mutation restores both high-affinity binding and signaling activity with 2′-O-methylated dsRNAs. Our work provides new structural insights into the mechanisms of host and viral immune evasion from RIG-I, explaining the complexity of cap structures over evolution.

Detailed, country‐specific epidemiological data are needed to characterize the burden of chronic hepatitis C virus (HCV) infection around the world. With new treatment options available, policy makers and public health officials must reconsider national strategies for infection control. In this study of 15 countries, published and unpublished data on HCV prevalence, viraemia, genotype, age and gender distribution, liver transplants and diagnosis and treatment rates were gathered from the literature and validated by expert consensus in each country. Viraemic prevalence in this study ranged from 0.2% in Iran and Lebanon to 4.2% in Pakistan. The largest viraemic populations were in Pakistan (7001000 cases) and Indonesia (3187000 cases). Injection drug use (IDU) and a historically unsafe blood supply were major risk factors in most countries. Diagnosis, treatment and liver transplant rates varied widely between countries. However, comparison across countries was difficult as the number of cases changes over time. Access to reliable data on measures such as these is critical for the development of future strategies to manage the disease burden.

The hepatitis C virus (HCV) epidemic was forecasted through 2030 for 15 countries in Europe, the Middle East and Asia, and the relative impact of two scenarios was considered: increased treatment efficacy while holding the annual number of treated patients constant and increased treatment efficacy and an increased annual number of treated patients. Increasing levels of diagnosis and treatment, in combination with improved treatment efficacy, were critical for achieving substantial reductions in disease burden. A 90% reduction in total HCV infections within 15 years is feasible in most countries studied, but it required a coordinated effort to introduce harm reduction programmes to reduce new infections, screening to identify those already infected and treatment with high cure rate therapies. This suggests that increased capacity for screening and treatment will be critical in many countries. Birth cohort screening is a helpful tool for maximizing resources. Among European countries, the majority of patients were born between 1940 and 1985. A wider range of birth cohorts was seen in the Middle East and Asia (between 1925 and 1995).

The total number, morbidity and mortality attributed to viraemic hepatitis C virus (HCV) infections change over time making it difficult to compare reported estimates from different years. Models were developed for 15 countries to quantify and characterize the viraemic population and forecast the changes in the infected population and the corresponding disease burden from 2014 to 2030. With the exception of Iceland, Iran, Latvia and Pakistan, the total number of viraemic HCV infections is expected to decline from 2014 to 2030, but the associated morbidity and mortality are expected to increase in all countries except for Japan and South Korea. In the latter two countries, mortality due to an ageing population will drive down prevalence, morbidity and mortality. On the other hand, both countries have already experienced a rapid increase in HCV‐related mortality and morbidity. HCV‐related morbidity and mortality are projected to increase between 2014 and 2030 in all other countries as result of an ageing HCV‐infected population. Thus, although the total number of HCV countries is expected to decline in most countries studied, the associated disease burden is expected to increase. The current treatment paradigm is inadequate if large reductions in HCV‐related morbidity and mortality are to be achieved.

Hepatitis C virus (HCV) is continuing to spread worldwide, adding three million new infections each year. Currently approved therapies are highly effective; however, access to them is limited due to the high cost of treatment. Therefore, a cost effective vaccine and alternative antivirals remain essential. HCV envelope glycoproteins, E1 and E2, heterodimerize on the virion surface and are the major determinant for virus pathogenicity and host immune response. Recent structural insights into amino-terminal domain of E1 and core of E2 have revealed unexpected folds not present in glycoproteins from related viruses. Here we discuss these structural findings with respect to their role in HCV entry and impact on potential vaccine design and new antivirals.

Hepatitis C Virus (HCV) infects 200 million individuals worldwide. Although several FDA approved drugs targeting the HCV serine protease and polymerase have shown promising results, there is a need for better drugs that are effective in treating a broader range of HCV genotypes and subtypes without being used in combination with interferon and/or ribavirin. Recently, two crystal structures of the core of the HCV E2 protein (E2c) have been determined, providing structural information that can now be used to target the E2 protein and develop drugs that disrupt the early stages of HCV infection by blocking E2’s interaction with different host factors. Using the E2c structure as a template, we have created a structural model of the E2 protein core (residues 421–645) that contains the three amino acid segments that are not present in either structure. Computational docking of a diverse library of 1,715 small molecules to this model led to the identification of a set of 34 ligands predicted to bind near conserved amino acid residues involved in the HCV E2: CD81 interaction. Surface plasmon resonance detection was used to screen the ligand set for binding to recombinant E2 protein, and the best binders were subsequently tested to identify compounds that inhibit the infection of Huh-7 cells by HCV. One compound, 281816, blocked E2 binding to CD81 and inhibited HCV infection in a genotype-independent manner with IC50’s ranging from 2.2 µM to 4.6 µM. 281816 blocked the early and late steps of cell-free HCV entry and also abrogated the cell-to-cell transmission of HCV. Collectively the results obtained with this new structural model of E2c suggest the development of small molecule inhibitors such as 281816 that target E2 and disrupt its interaction with CD81 may provide a new paradigm for HCV treatment.

Gilead Sciences The disease burden of hepatitis C virus (HCV) is expected to increase as the infected population ages. A modelling approach was used to estimate the total number of viremic infections, diagnosed, treated and new infections in 2013. in addition, the model was used to estimate the change in the total number of HCV infections, the disease progression and mortality in 2013-2030. Finally, expert panel consensus was used to capture current treatment practices in each country. Using today's treatment paradigm, the total number of HCV infections is projected to decline or remain flat in all countries studied. However, in the same time period, the number of individuals with late-stage liver disease is projected to increase. This study concluded that the current treatment rate and efficacy are not sufficient to manage the disease burden of HCV. Thus, alternative strategies are required to keep the number of HCV individuals with advanced liver disease and liver-related deaths from increasing. Ctr Dis Anal, Louisville, CO 80027 USA Natl Liver Inst, Menoufia, Egypt JW Goethe Univ Hosp, Frankfurt, Germany Univ Calgary, Liver Unit, Div Gastroenterol & Hepatol, Calgary, AB, Canada Ankara Univ, Dept Gastroenterol, Sch Med, TR-06100 Ankara, Turkey Hosp Santo Antonio Capuchos, Dept Gastroenterol, Ctr Hosp Lisboa Cent, Lisbon, Portugal Med Univ Innsbruck, A-6020 Innsbruck, Austria Univ São Paulo, Sch Med, São Paulo, Brazil Hop Henri Mondor, Serv Hepatogastroenterol, F-94010 Creteil, France Adv Tech Hlth Serv Res TAISS, Madrid, Spain Ege Univ, Izmir, Turkey Karolinska Inst, Dept Med Huddinge, Stockholm, Sweden Karolinska Univ Hosp, Dept Gastroenterol & Hepatol Infect Dis, Stockholm, Sweden Ankara Univ, TR-06100 Ankara, Turkey Univ Leipzig, D-04109 Leipzig, Germany Osped Cantonale, Dept Gastroenterol, Bellinzona, Switzerland Univ Montreal, Dept Med, Liver Unit, Montreal, PQ H3C 3J7, Canada Fed Univ State Rio de Janeiro, Dept Gastroenterol, Rio de Janeiro, Brazil Arud Ctr Addict Med, Zurich, Switzerland Hosp Valle de Hebron, CIBERehd, Barcelona, Spain Hosp Puerta Hierro, Madrid, Spain Univ Fed Rio Grande do Sul, Hosp Clin, Porto Alegre, RS, Brazil Odense Univ Hosp, Dept Infect Dis, DK-5000 Odense, Denmark Reg Hosp Hovedstaden, Copenhagen, Denmark Univ Fed Rio de Janeiro, Dept Clin Med, Rio de Janeiro, Brazil Univ Plymouth, Peninsula Sch Med & Dent, Plymouth PL4 8AA, Devon, England Univ New S Wales, Kirby Inst, Sydney, NSW, Australia Cairo Univ, Cairo, Egypt Orebro Univ Hosp, Dept Infect Dis, Orebro, Sweden Univ Orebro, Sch Hlth & Med Sci, SE-70182 Orebro, Sweden Ain Shams Univ, Cairo, Egypt Dokuz Eylul Univ, Izmir, Turkey Karolinska Inst, Karolinska Univ Hosp, Dept Med Huddinge, Infect Dis Unit, Stockholm, Sweden Exigo Consultores, Alhos Vedros, Portugal Universidade Federal de São Paulo, Div Gastroenterol, São Paulo, Brazil Universidade Federal de São Paulo, Div Infect Dis, São Paulo, Brazil Inst Clin & Expt Med, Dept Hepatogastroenterol, Prague, Czech Republic Hosp Carlos III, CIBERehd, Madrid, Spain Univ Copenhagen, Copenhagen, Denmark Direccao Geral Saude, Lisbon, Portugal Univ Estadual Campinas, Disciplina Doencas Infecciosas, Dept Clin Med, Grp Estudo Hepatites,Fac Ciencias Med,UNICAMP, São Paulo, Brazil Wilhelminenspital Stadt Wien, Dept Internal Med 4, Vienna, Austria Univ São Paulo, Sch Med, Div Gastroenterol & Hepatol, São Paulo, Brazil Med Univ Vienna, Div Gastroenterol & Hepatol, Dept Internal Med 3, Vienna, Austria Masaryk Univ, Univ Hosp Brno, Clin Infect Dis, Brno, Czech Republic Univ Manitoba, Dept Internal Med, Sect Hepatol, Winnipeg, MB, Canada Hlth Sci Ctr, Viral Hepatitis Invest Unit, Winnipeg, MB, Canada European Liver Patients Assoc, St Truiden, Belgium Istanbul Univ, Istanbul, Turkey Univ British Columbia, British Columbia Ctr Dis Control, Vancouver, BC V5Z 1M9, Canada Aalborg Univ Hosp, Dept Med Gastroenterol, Aalborg, Denmark Aalborg Univ Hosp, Sect Mol Diagnost, Aalborg, Denmark Katholieke Univ Leuven, Univ Hosp Leuven, Louvain, Belgium Hosp Santa Maria, Dept Gastroenterol, Ctr Hosp Lisboa Norte, Lisbon, Portugal Univ Western Ontario, Div Gastroenterol, London, ON, Canada Univ Dusseldorf, Dusseldorf, Germany Univ Libre Brussels, Erasme Univ Hosp, Brussels, Belgium Univ Hosp, Div Gastroenterol & Hepatol, Geneva, Switzerland Univ Hosp, Div Clin Pathol, Geneva, Switzerland Natl Inst Publ Hlth, Natl Reference Lab Hepatitis, Prague, Czech Republic Univ Southhampton, Southampton, England Dalhousie Univ & Hepatol Serv, Capital Dist Hlth Author, Queen Elizabeth II Hlth Sci Ctr, Dept Med, Halifax, NS, Canada Dalhousie Univ & Hepatol Serv, Capital Dist Hlth Author, Queen Elizabeth II Hlth Sci Ctr, Dept Surg, Halifax, NS, Canada Univ British Columbia, Dept Gastroenterol, Vancouver, BC V5Z 1M9, Canada Assembleia Republ, Lisbon, Portugal Alfred Hosp, Melbourne, Vic, Australia Monash Univ, Melbourne, Vic 3004, Australia UCL, Div Med, UCL Inst Liver & Digest Hlth, London, England Hop Henri Mondor, Dept Sante Publ, F-94010 Creteil, France Nottingham Univ Hosp NHS Trust, Nottingham, England Biomed Res Unit, Nottingham, England Ctr Hosp Porto, Dept Infect Dis, Oporto, Portugal Cantonal Hosp St Gallen, Div Gastroenterol & Hepatol, St Gallen, Switzerland Univ Toronto, Toronto Gen Hosp, Univ Hlth Network, Toronto, ON M5G 1L7, Canada Egyptian Liver Res Inst & Hosp, Dakahliah, Egypt Monash Hlth, Melbourne, Vic, Australia Catholic Univ Louvain, Clin Univ St Luc, Brussels, Belgium Med Univ Graz, Div Gastroenterol & Hepatol, Dept Internal Med, Graz, Austria St Vincents Hosp, Dept Gastroenterol, Melbourne, Vic, Australia Univ Melbourne, Melbourne, Vic, Australia Charles Univ Prague, Dept Internal Med, Fac Med 1, Prague, Czech Republic Cent Mil Hosp, Prague, Czech Republic Univ Antwerp, B-2020 Antwerp, Belgium Deutsch Leberhilfe eV, Cologne, Germany Ghent Univ Hosp, Ghent, Belgium Univ Ghent, Belgium Hasselt Univ, Dept Hlth Econ & Patient Safety, Diepenbeek, Belgium Hannover Med Sch, Dept Gastroenterol Hepatol & Endocrinol, Hannover, Germany Copenhagen Univ Hosp, Hvidovre, Denmark Univ New S Wales, St George Hosp Clin Sch Med, Sydney, NSW, Australia Univ New S Wales, Sch Med Sci, Sydney, NSW, Australia Univ Zurich Hosp, Swiss HPB Hepatopancreatobiliary Ctr, CH-8091 Zurich, Switzerland Univ Zurich Hosp, Dept Gastroenterol & Hepatol, CH-8091 Zurich, Switzerland Universidade Federal de São Paulo, Div Gastroenterol, São Paulo, Brazil Universidade Federal de São Paulo, Div Infect Dis, São Paulo, Brazil Web of Science

The crystal structure of the core domain of the hepatitis C virus surface glycoprotein E2 has been solved; the structure shows that, contrary to expectation, E2 is unlikely to be the viral fusion protein. There is currently no vaccine against hepatitis C virus, so it is important to learn more about the processes by which the virus establishes infection. Joseph Marcotrigiano and colleagues solve the crystal structure of the core domain of the hepatitis C virus surface glycoprotein E2. The structure shows that, contrary to expectation, E2 is unlikely to be the viral fusion protein. This work helps to clarify the role of E2 and the mechanism of hepatitis C virus entry. Hepatitis C virus (HCV) is a significant public health concern with approximately 160 million people infected worldwide1. HCV infection often results in chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. No vaccine is available and current therapies are effective against some, but not all, genotypes. HCV is an enveloped virus with two surface glycoproteins (E1 and E2). E2 binds to the host cell through interactions with scavenger receptor class B type I (SR-BI) and CD81, and serves as a target for neutralizing antibodies2,3,4. Little is known about the molecular mechanism that mediates cell entry and membrane fusion, although E2 is predicted to be a class II viral fusion protein. Here we describe the structure of the E2 core domain in complex with an antigen-binding fragment (Fab) at 2.4 A resolution. The E2 core has a compact, globular domain structure, consisting mostly of β-strands and random coil with two small α-helices. The strands are arranged in two, perpendicular sheets (A and B), which are held together by an extensive hydrophobic core and disulphide bonds. Sheet A has an IgG-like fold that is commonly found in viral and cellular proteins, whereas sheet B represents a novel fold. Solution-based studies demonstrate that the full-length E2 ectodomain has a similar globular architecture and does not undergo significant conformational or oligomeric rearrangements on exposure to low pH. Thus, the IgG-like fold is the only feature that E2 shares with class II membrane fusion proteins. These results provide unprecedented insights into HCV entry and will assist in developing an HCV vaccine and new inhibitors.

New knowledge on the treatment of Hepatitis C is accruing at an extraordinarily rapid rate and my aim in this editorial based on a talk I gave at the Pakistan Society of Gastroenterology Annual International Congress this year, is to outline how patients may be treated both currently and over the next two to three years.

Intercellular adhesion molecule 1 (ICAM-1) mediates binding and entry of major group human rhinoviruses (HRVs). Whereas the entry pathway of minor group HRVs has been studied in detail and is comparatively well understood, the pathway taken by major group HRVs is largely unknown. Use of immunofluorescence microscopy, colocalization with specific endocytic markers, dominant negative mutants, and pharmacological inhibitors allowed us to demonstrate that the major group virus HRV14 enters rhabdomyosarcoma cells transfected to express human ICAM-1 in a clathrin-, caveolin-, and flotillin-independent manner. Electron microscopy revealed that many virions accumulated in long tubular structures, easily distinguishable from clathrin-coated pits and caveolae. Virus entry was strongly sensitive to the Na+/H+ion exchange inhibitor amiloride and moderately sensitive to cytochalasin D. Thus, cellular uptake of HRV14 occurs via a pathway exhibiting some, but not all, characteristics of macropinocytosis and is similar to that recently described for adenovirus 3 entry via αvintegrin/CD46 in HeLa cells.

Minor group human rhinoviruses (HRVs) bind three members of the low-density lipoprotein receptor (LDLR) family: LDLR proper, very-LDLR (VLDLR) and LDLR-related protein (LRP). Whereas ICAM-1, the receptor of major group HRVs actively contributes to viral uncoating, LDLRs are rather considered passive vehicles for cargo delivery to the low-pH environment of endosomes. Since the Tyr-Trp-Thr-Asp β-propeller domain of LDLR has been shown to be involved in the dissociation of bound LDL via intramolecular competition at low pH, we studied whether it also plays a role in HRV infection. Human cell lines deficient in LDLR family proteins are not available. Therefore, we used CHO -ldla7 cells that lack endogenous LDLR. These were stably transfected to express either wild-type (wt) human LDLR or a mutant with a deletion of the β-propeller. When HRV2 was attached to the propeller-negative LDLR, a lower pH was required for conversion to subviral particles than when attached to wt LDLR. This indicates that high-avidity receptor binding maintains the virus in its native conformation. HRV2 internalization directed the mutant LDLR but not wt LDLR to lysosomes, resulting in reduced plasma membrane expression of propeller-negative LDLR. Infection assays using a CHO-adapted HRV2 variant showed a delay in intracellular viral conversion and de novo viral synthesis in cells expressing the truncated LDLR. Our data indicate that the β-propeller attenuates the virus-stabilizing effect of LDLR binding and thereby facilitates RNA release from endosomes, resulting in the enhancement of infection. This is a nice example of a virus exploiting high-avidity multimodule receptor binding with an intrinsic release mechanism.

Major group HRVs bind intercellular adhesion molecule 1 and minor group HRVs bind members of the low-density lipoprotein receptor (LDLR) family for cell entry. Whereas the former share common sequence motives in their viral capsid proteins (VPs), in the latter only a lysine residue within the binding epitope in VP1 is conserved; this lysine is also present in “K-type” major group HRVs that fail to use LDLR for infection. By using the available sequences three-dimensional models of VP1 of all HRVs were built and binding energies, with respect to module 3 of the very-low-density lipoprotein receptor, were calculated. Based on the predicted affinities K-type HRVs and minor group HRVs were correctly classified.