Serum lipids and chronic hepatitis C genotype 4: interaction and significance

Background &; aim. Metabolic abnormalities are common in chronic hepatitis C infection (CHC). However, the genotypic differences of these disarrangements in patients infected with CHC genotype 4 (HCV-4) and its association with liver histology and viral loads remain unknown.Material and methods. We consecutively enrolled 183 HCV-4 patients and 106 healthy matched controls; to compare metabolic profiles and assess pattern of association of HCV RNA levels as well as histological factors with the serum lipid profile.Results. HCV-4 infection is associated with higher homeostasis model assessment of insulin resistance (HOMA-IR) index, despite that, a favourable lipid pattern, consisting of an elevation in HDLC and a reduction in serum cholesterol (TC), LDL-C and triglyceride (TG) levels, in comparison with normal matched adults. Significant fibrosis was independently associated with HOMA-IR, portal/periportal inflammation grade, serum cholesterol and age. Univariate association was elucidated between lower LDL-C and TC and Metavir activity score and between higher TG and TC and steatosis. In multivariate analysis, severe hepatitis activity, milder hepatic fibrosis, and triglyceride levels are associated with higher HCV RNA levels.Conclusion. HCV-4 is associated with wide metabolic changes. A proportional relationship is found between serum lipid profiles and hepatitis C viral load and liver histology in patients with HCV-4.     

Association of microRNAs genes polymorphisms with rheumatoid arthritis in Egyptian female patients

ObjectiveTo investigate whether miRNA-499 (rs3746444) and miRNA-146a (rs2910164) genes polymorphisms are independent factors for rheumatoid arthritis (RA) in Egyptians, and whether they influence disease severity and activity.MethodsTwo hundred and seventeen RA patients and 245 healthy controls were enrolled in this study. Polymorphisms of miRNA-146a and miRNA-499 genes were detected using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP).ResultsThe miRNA-499 CT genotype was an independent factor of RA. The miRNA-499 CT, CC genotypes and C allele frequencies were significantly increased in erosive RA group. Moreover, the heterozygote CT had more severe and more active form of the disease compared with homozygote CC or TT. However, we did not find any significant association of miRNA-146a polymorphism with RA risk, severity, and activity.ConclusionThe miRNA-499 polymorphism is an independent factor of RA, and influences disease severity and activity.     

Multicompartmental Analysis of Cholesterol Metabolism in Man: CHARACTERIZATION OF THE HEPATIC BILE ACID AND BILIARY CHOLESTEROL PRECURSOR SITES

The present report has presented the first clear evidence in man for the existence of specific hepatic cholesterol precursor sites associated with the formation and secretion of bile acids and biliary cholesterol. These hepatic compartments derive virtually all their cholesterol from newly synthesized and lipoprotein free cholesterol. The model which is presented was formulated on current concepts of cholesterol metabolism in man and is concerned, at this initial stage, with the elucidation of the bile acid and biliary cholesterol compartments. The complexity of cholesterol metabolism in man necessitated an initial approach that would minimize the number of inputs of cholesterol into the system, allow for the sampling of several cholesterol compartments, and permit the simultaneous labeling of newly synthesized cholesterol and preformed cholesterol. To achieve these objectives, we studied the patient with a total bile fistula. Six patients were administered simultaneously pulse injections of labeled mevalonic acid and [(14)C]cholesterol. The qualitative features of the specific activity time course curves after labeled mevalonic acid revealed no precursor-product relationship between bile acid, biliary cholesterol, and plasma free cholesterol. The peak specific activity of the bile acids was reached in approximately 100 min and was higher than the biliary cholesterol, which was higher than the plasma free cholesterol. The plasma free cholesterol specific activity became higher than the other lipids after 12 h and remained higher throughout the period of study. Similar related observations were made with [(14)C]cholesterol. The data were then subjected to simulation analysis and modeling using the SAAM-27 computer program. Computer least-square fits of the data were obtained after the model was evolved. During the model development, the least number of compartments and transport pathways were introduced consistent with a good fit of the data. Of particular importance was the constraint that the model fit the data obtained from both [(14)C]cholesterol and labeled mevalonic acid. The same parameter values were used to fit the data from both tracers. The fluxes arrived at in the model indicate that 31% and 20%, respectively, of the cholesterol input into the bile acid and biliary cholesterol precursor sites were derived directly from the newly synthesized hepatic cholesterol. The remainder had its origin predominantly from lipoprotein free cholesterol. Plasma esterified cholesterol (as free) made a small contribution (11%) to the bile acid compartment. Similarly, 10% of the biliary cholesterol arose from an unknown hepatic site.The present report has provided the basis for a new procedure for studying in vivo cholesterol metabolism in man. Examination of the derived cholesterol flux rates between the compartments suggests the presence of an important mechanism regulating the partitioning of lipoprotein free cholesterol between the bile acid and biliary cholesterol precursor sites. Aberrations in the proportioning of precursor cholesterol between these sites could be a causative factor precipitating the excessive secretion of biliary cholesterol and the production of lithogenic bile.     

Abordaje laparoscópico de la acalasia. Resultados clínicos,de calidad de vida y funcionales a largo plazo

Introduction

Laparoscopic Heller myotomy has become the gold standard procedure for patients with achalasia. This study evaluates the clinical status, quality of life, and functional outcomes after laparoscopic Heller myotomy.

Material and methods

We analyzed patients who underwent laparoscopic Heller myotomy with an associated anti-reflux procedure from October 1998 to December 2010. Before surgery, we administered a clinical questionnaire and as of 2002, we also evaluated quality of life using a specific questionnaire (GIQLI). In 2011, we performed a follow up for all available patients. We administered the same clinical questionnaire and quality of life test as before surgery and performed manometry and 24-hour pH monitoring. According to the length of follow up, patients were divided into 3 groups. Group 1 with a follow-up between 6 and 47 months; group 2 follow-up between 48 and 119 months, and group 3 with a follow-up of more than 120 months). Moreover, 27 patients had already been evaluated with this same protocol in 2003. Pre- and postoperative data were compared for the 3 groups and for patients who completed follow up in 2003 and 2011.

Results

Ninety-five patients underwent laparoscopic Heller myotomy. Seventy-six (80%) were available for follow-up. Mean follow-up was 56 months (range 6-143). Global improvement in dysphagia was 89%. Total DeMeester score decreased in the 3 groups. GIQLI scores improved after surgery, reaching normal values. Manometric determinations showed normal LES pressures after myotomy in the 3 groups. Ten percent of overall 24-hour pH monitoring was abnormal. The group of patients followed up in 2003 and in 2011 showed no impairment in the variables studied in the long term.

Conclusions

Long-term follow up of the laparoscopic approach to achalasia showed good results concerning clinical status and quality of life, with normal sphincteric pressures and a low incidence of gastroesophageal reflux.     

A qualitative study of stakeholder views and experiences of minor ailment services in the United Kingdom

BackgroundAn international strategy designed to promote access to primary care is the utilisation of community pharmacy to deliver structured minor ailment services (MASs). An understanding of key implementation features of MASs will support effective service delivery and implementation, promote MAS viability, sustainability and overall improvement.AimThe aim of this study is to explore the views and experiences of a range of stakeholders concerning the implementation of MASs in the United Kingdom.MethodsA qualitative approach was used to obtain data. Participants were recruited using purposeful and snowball sampling. Stakeholders from five different regions were included. Using the digital recordings of the interviews, thematic content analysis was undertaken.ResultsThirty-three participants agreed to be interviewed. Twenty-nine semi-structured interviews were conducted. Thematic content analysis yielded three major themes, including (1) benefits of MASs, (2) structural challenges associated with MAS design and (3) other implementation factors associated with MAS delivery. Stakeholders recognised the positive impact of the service to improve patient access and care, promote efficiencies, and promote the professional role of the pharmacist. Nevertheless barriers do exist to service delivery and implementation. Stakeholders identified the need to potentially increase the population groups served by MASs, increase the conditions treated and widen their formulary lists. Similarly, marketing strategies needed to be improved to enhance consumer awareness. Stakeholders presented mixed views about whether pharmacists needed to complete clinical training and the need to increase pharmacist's remuneration. In addition the level of healthcare collaboration needed to improve.ConclusionSeveral concepts emerged from the investigation to facilitate service delivery. Barriers to service implementation had a variable impact on implementation. Service delivery should function to meet all stakeholder needs and can be achieved through stakeholder collaboration. However, improved marketing to promote consumer awareness together with better collaborative processes can potentially improve MAS implementation.     

Reliability of long vs short COI markers in identification of forensically important flies

Aim

To compare the reliability of short and long cytochrome oxidase I gene fragment (COI) in identification of forensically important Diptera from Egypt and China.

Methods

We analyzed 50 specimens belonging to 18 species. The two investigated markers were amplified by polymerase chain reaction (PCR) followed by direct sequencing. Nucleotide sequence divergences were calculated using the Kimura two-parameter (K2P) distance model and neighbor-joining (NJ) phylogenetic trees.

Results

Although both tested fragments showed an overlap between intra and interspecific variations, long marker had greater completeness of monophyletic separation with high bootstrap support. Moreover, NJ tree based on the long fragment clustered species more in accordance with their taxonomic classification than that based on the short fragment.

Conclusion

In dipterous identification, it is recommended to use the long COI marker due to its greater reliability and safety.Necrophagous insects can serve as a valuable source of information for estimation of minimum post-mortem interval (PMI) in legal medicine. Most suitable for forensic purposes are species from the order Diptera (eg, Calliphoridae, Muscidae, and Sarcophagidae) (1-4). In PMI estimation, an important initial step is correct identification of these insects, which may be difficult by using the traditional morphology-based approach (5,6), because several forensically important fly species can hardly be distinguished morphologically (7-9). The limitations of morphological method can be overcome by gene sequences analysis, a fast and accurate method of species identification. Molecular analysis requires small tissue samples and is relatively insensitive to preservation conditions (1,10). Different mitochondrial (mt) and nuclear (nu) DNA markers are investigated as forensic tools. However, mtDNA is preferred because it can be easily extracted even from small or degraded samples (10). In addition, because of its strictly maternal inheritance and lack of genetic recombination, mtDNA haplotype is a good candidate for evolutionary and population genetics study.Mitochondrial cytochrome c oxidase subunit I (COI) sequences are a rapid and powerful tool for accurate identification of species across various taxa (7,11-14). Although COI has been extensively studied by forensic entomologists, resulting in a vast amount of DNA data, there is little agreement as to which portion of the gene needs to be sequenced. Although the 5′ end of COI is also the site of the proposed universal animal DNA “barcode” (11) and it has been successfully used in the identification of many blowfly species (12), this approach cannot identify some closely related species (12,15). Therefore, to optimize discrimination power between closely related species some authors suggested multi-gene approach (16,17). Surprisingly, a recent study using this approach revealed that phylogenetic tree based on COI fragment was similar to that based on 3 different gene fragments (16).Fragments of the COI sequence that show low sequence divergence within species but high divergences among species can be employed as taxon “barcodes,” and unknown samples can be accurately grouped to species with reference sequences of the “barcode library” (14,18,19). Therefore, it is paramount to evaluate not only discrimination power of these COI fragments between closely related species but also between species belonging to more than one family, because in a database an unknown sample will be compared to all reference samples. In the absence of an appropriate reference sample, unknown samples will simply group with the most closely matched reference sample (20). Thus, it is important to confirm that the investigated marker will not only be correctly assigned to a species but also that it will be in accordance with the traditional morphological classification. Therefore, we evaluated the discrimination power of the short (272-bp) COI fragment in the identification of the most forensically relevant flies (Calliphoridae, Sarcophagidae, and Muscidae) originating from Egypt and China in comparison to the long (1173-bp) COI fragment, and aimed to gather genetic data on common forensically important Diptera.     

Relationships between TIMP-1, CAIX,and clinical outcomes in Egyptian breast cancer

Serum levels of carbonic anhydrase IX (CAIX) and the tissue inhibitor of metalloproteinase 1 (TIMP-1) have been demonstrated to be associated with clinical outcomes. Elevated TIMP-1 levels have been associated with a poor prognosis of breast cancer. Therefore, this study was performed to explore the relationships between serum CAIX, TIMP-1, and clinical outcomes. One hundred and five female breast cancer patients, who do not receive adjuvant therapy at Mansoura University Hospital were recruited with informed consent for this study. Preoperative serum levels of CAIX and TIMP-1 were measured using an enzyme-linked immunosorbent assay. Among the breast cancer patients, 16.1 and 20.9 % had elevated TIMP-1 and CAIX l levels, respectively. In addition, 13.3 % had elevated both CAIX and TIMP-1 levels. Elevated serum levels CAIX was significantly associated with advanced histological grade, estrogen receptor status and progesterone receptor status. Moreover, Elevated TIMP-1 was significantly associated with number of tumor-positive lymph nodes and advanced histological grade. In Kaplan–Meier analysis, the disease-free survival (DFS) was lower in patients with high serum levels of CAIX and TIMP-1 compared to patients with normal serum concentration of CAIX and TIMP-1. Including TIMP-1, CAIX, and established prognostic factors in the multivariate analysis, the presence of elevated CAIX and TIMP-1 levels remained independent predictors of overall survival time (OS). A high serum TIMP-1 and CAIX levels were significantly associated with poorer DFS and OS among Egyptian primary breast cancer patients with elevated serum levels of CAIX and TIMP-1.     

In silico identification of potential SARS COV-2 2′-O-methyltransferase inhibitor: fragment-based screening approach and MM-PBSA calculations

In the present era, there are many efforts trying to face the emerging and successive waves of the COVID-19 pandemic. This has led to considering new and unusual targets for SARS CoV-2. 2′-O-Methyltransferase (nsp16) is a key and attractive target in the SARS CoV-2 life cycle since it is responsible for the viral RNA protection via a cap formation process. In this study, we propose a new potential inhibitor for SARS COV-2 2′-O-methyltransferase (nsp16). A fragment library was screened against the co-crystal structure of the SARS COV-2 2′-O-methyltransferase complexed with Sinefungin (nsp16 – PDB ID: 6WKQ), and consequently the best proposed fragments were linked via a de novo approach to build molecule AP-20. Molecule AP-20 displayed a superior docking score to Sinefungin and reproduced the key interactions in the binding site of 2′-O-methyltransferase. Three molecular dynamic simulations of the 2′-O-methyltransferase apo structure and its complexed forms with AP-20 and Sinefungin were performed for 150 nano-seconds to provide insights on the dynamic nature of such setups and to assess the stability of the proposed AP-20/enzyme complex. AP-20/enzyme complex demonstrated better stability for the ligand–enzyme complex compared to Sinefungin in a respective setup. Furthermore, MM-PBSA binding free energy calculations showed a better profile for AP-20/enzyme complex compared to Sinefungin/enzyme complex emphasizing the potential inhibitory effect of AP-20 on SARS COV-2 2′-O-methyltransferase. We endorse our designed molecule AP-20 to be further explored via experimental evaluations to confront the spread of the emerging COVID-19. Also, in silico ADME profiling has ascribed to AP-20 an excellent safety and metabolic stability profile.

The identification of AP-20 as a potential SARS COV-2 2′-O-methyltransferase inhibitor: fragment-based screening approach and MM-PBSA calculations.