SUMMARY PCR has revolutionized the field of infectious disease diagnosis. To overcome the inherent disadvantage of cost and to improve the diagnostic capacity of the test, multiplex PCR, a variant of the test in which more than one target sequence is amplified using more than one pair of primers, has been developed. Multiplex PCRs to detect viral, bacterial, and/or other infectious agents in one reaction tube have been described. Early studies highlighted the obstacles that can jeopardize the production of sensitive and specific multiplex assays, but more recent studies have provided systematic protocols and technical improvements for simple test design. The most useful of these are the empirical choice of oligonucleotide primers and the use of hot start-based PCR methodology. These advances along with others to enhance sensitivity and specificity and to facilitate automation have resulted in the appearance of numerous publications regarding the application of multiplex PCR in the diagnosis of infectious agents, especially those which target viral nucleic acids. This article reviews the principles, optimization, and application of multiplex PCR for the detection of viruses of clinical and epidemiological importance.
The importance of activins and follistatin in liver diseases has recently emerged. The aim of the present study was to measure the influence of chronic infection with viral hepatitis C (CHC) genotype 1 and 4 on serum levels of activin-A, activin-B and follistatin, and to determine their correlations with viral load, liver damage, interleukin-6 (IL-6) and tumour necrosis factor (TNF)-α. Sera samples collected from 20 male and 20 female treatment naïve CHC genotype 1 and 4 Saudi patients (ten males and ten females for each genotype), and 40 gender- and age-matched healthy participants were analysed for activin-A, activin-B and follistatin using enzyme-linked immunosorbent assay and their levels were correlated with IL-6, TNF-α, viral load and AST platelet ratio index (APRI). Serum activin-A, activin-B, IL-6 and TNF-α were significantly increased, while serum follistatin was significantly decreased, in both genders of CHC patients compared with control subjects, In both viral genotypes, activin-A was strongly and positively correlated with the viral load, APRI, IL-6 and TNF-α, and negatively with albumin (P < 0.01). Activin-B showed the same correlations of activin-A only in CHC genotype 1 patients, but it was weaker than activin-A. No correlation was detected with follistatin. Serum activins, particularly activin-A, and follistatin are significantly altered by CHC genotype 1 and 4. This dysregulation of activins/follistatin axis may be associated with viral replication, host immune response and liver injury. Further studies are needed to illustrate the definite role(s) and clinical value of activins and follistatin in CHC.
To measure the prevalence of cytomegalovirus (CMV) infection in ectopic pregnancy (EP) and its effect on the expression of inducible and endothelial nitric oxide synthases (iNOS, eNOS) by Fallopian tubes (FT) bearing an EP. This was a prospective case-control study. Blood and tubal samples were collected from 84 Eps and 51 controls (20 total abdominal hysterectomy (TAH) during the luteal phase and another 31 tubal ligations). CMV IgM and IgG antibodies were measured by ELISA, and an IVD CE PCR kit was used to detect CMV in the FTs. iNOS and eNOS were measured by immunohistochemistry and quantitative RT-PCR in FTs obtained from CMV-positive EP (n = 12), and the results were compared with those obtained from CMV-negative EP (n = 11) and TAH (n = 8). The frequencies of CMV IgM (51.2 % vs 17.6 %), IgG (77.4 % vs 52.9 %) or both antibodies (41.6 % vs 11.7 %) were significantly higher in EP compared with control. CMV was more common by PCR in FTs from EP (21.4 %) than controls (5.9 %). Twelve women from the PCR positive EP cases (66.6 %) were also simultaneously positive for both CMV IgM & IgG antibodies and had higher expression of eNOS and iNOS at the protein and gene levels compared with negative EP and TAH. Tubal infection with CMV may lead to EP by increasing the production of endothelial and inducible NOS by the FT epithelial cells. Further studies are required to illustrate the role of CMV in the pathogenesis of EP.
Table of contents O1 Regulation of genes by telomere length over long distances Jerry W. Shay O2 The microtubule destabilizer KIF2A regulates the postnatal establishment of neuronal circuits in addition to prenatal cell survival, cell migration, and axon elongation, and its loss leading to malformation of cortical development and severe epilepsy Noriko Homma, Ruyun Zhou, Muhammad Imran Naseer, Adeel G. Chaudhary, Mohammed Al-Qahtani, Nobutaka Hirokawa O3 Integration of metagenomics and metabolomics in gut microbiome research Maryam Goudarzi, Albert J. Fornace Jr. O4 A unique integrated system to discern pathogenesis of central nervous system tumors Saleh Baeesa, Deema Hussain, Mohammed Bangash, Fahad Alghamdi, Hans-Juergen Schulten, Angel Carracedo, Ishaq Khan, Hanadi Qashqari, Nawal Madkhali, Mohamad Saka, Kulvinder S. Saini, Awatif Jamal, Jaudah Al-Maghrabi, Adel Abuzenadah, Adeel Chaudhary, Mohammed Al Qahtani, Ghazi Damanhouri O5 RPL27A is a target of miR-595 and deficiency contributes to ribosomal dysgenesis Heba Alkhatabi O6 Next generation DNA sequencing panels for haemostatic and platelet disorders and for Fanconi anaemia in routine diagnostic service Anne Goodeve, Laura Crookes, Nikolas Niksic, Nicholas Beauchamp O7 Targeted sequencing panels and their utilization in personalized medicine Adel M. Abuzenadah O8 International biobanking in the era of precision medicine Jim Vaught O9 Biobank and biodata for clinical and forensic applications Bruce Budowle, Mourad Assidi, Abdelbaset Buhmeida O10 Tissue microarray technique: a powerful adjunct tool for molecular profiling of solid tumors Jaudah Al-Maghrabi O11 The CEGMR biobanking unit: achievements, challenges and future plans Abdelbaset Buhmeida, Mourad Assidi, Leena Merdad O12 Phylomedicine of tumors Sudhir Kumar, Sayaka Miura, Karen Gomez O13 Clinical implementation of pharmacogenomics for colorectal cancer treatment Angel Carracedo, Mahmood Rasool O14 From association to causality: translation of GWAS findings for genomic medicine Ahmed Rebai O15 E-GRASP: an interactive database and web application for efficient analysis of disease-associated genetic information Sajjad Karim, Hend F Nour Eldin, Heba Abusamra, Elham M Alhathli, Nada Salem, Mohammed H Al-Qahtani, Sudhir Kumar O16 The supercomputer facility “AZIZ” at KAU: utility and future prospects Hossam Faheem O17 New research into the causes of male infertility Ashok Agarwa O18 The Klinefelter syndrome: recent progress in pathophysiology and management Eberhard Nieschlag, Joachim Wistuba, Oliver S. Damm, Mohd A. Beg, Taha A. Abdel-Meguid, Hisham A. Mosli, Osama S. Bajouh, Adel M. Abuzenadah, Mohammed H. Al-Q...
BackgroundActivin-A and follistatin regulate the liver and the immune system.AimsTo measure the effects of treatment with pegylated-interferon-α (Peg-IFN-α) and ribavirin on the concentrations of mature activin-A and follistatin in serum and liver tissue homogenates in rats.MethodsA total of 28 male Wistar rats were divided equally into four groups as follow: ‘Control group’ (n = 7), ‘PEG only group’ consisted of those that only received a weekly injection of Peg-IFN-α (6 µg/rat) for 4 weeks, ‘RBV only group’ received ribavirin only (4 mg/rat/day) orally for 35 days and the last group received both Peg-IFN-α and ribavirin ‘PEG & RBV group’. The concentrations of candidate proteins in serum and liver samples were measured using ELISA.ResultsPegylated-interferon-α decreased activin-A and increased follistatin significantly in serum and liver of ‘PEG only’ and ‘PEG & RBV’ groups compared with the ‘Control’ and ‘RBV only’ groups (P < 0.05). There was no significant difference between the ‘RBV only’ and ‘Control’ groups (P > 0.05) in the concentrations of candidate proteins. A significant positive correlations between serum and liver activin-A (r = 0.727; P = 0.02 × 10−3) and follistatin (r = 0.540; P = 0.01) was also detected.ConclusionPegylated-interferon-α modulates the production of activin-A and follistatin by the liver, which is reflected and can be detected at the serum level. Further studies are needed to explore the role of Peg-IFN-α based therapy on the production of activins and follistatin by the liver and immune cells.
ÖZETAmaç: Kronik hepatit C (KHC) ve Peg-interferon-alfa (Peg-INF-α) serum aktivinleri ve follistatini düzenler ve tiroid hastalıkları (TH) ile birliktedir. Bu çalışmanın amacı KHC'nin indüklediği TH sıklığını belirlemek ve karaciğer hasarı, serum activin-A ve follistatin'in tiroid fonksiyonları ve tiroid otoantikorları ile ilişkisini araştırmaktır.Yöntemler: Bu çalışma kesitsel olup, 3 gruba ayrılan 132 KHC'li hastadan serumları alınmıştır: Tedavisiz 56 hasta, 24 haftalık Peg-INF-α tedavisi almış 30 hasta ve Peg-INF-α tedavisi tamamlanmış 46 hasta. Tiroid stimülan hormon (TSH), serbest tiroksin (FT4) ve tiroid otoantikorları, serum activin-A ve follistatin düzeyleri ELISA yönte-miyle ölçüldü. Bulgular:Hastaların %15'inde (n=20) tiroid hastalığı saptandı, bunların çoğunluğu (%80) otoimmün tiroidit olup, kadınlarda daha sık (%70) idi. TSH reseptör antikorları (TSHR-Abs), diğer antikorlardan anlamlı yüksek bulundu (p<0,05) ve 24 haftalık tedavi grubunda anlamlı artmıştı (p<0,05). TSH ve FT4 karaciğer enzimleri ile anlamlı kö-rele idi (p<0,05). Tiroid hastalığı olanlarla ötiroid olanlar arasında activin-A ve follistatin değerleri bakımından anlamlı fark yoktu (p>0,05). Ancak TSHR-Abs ile follistatin, activin-A ve activi/follistatin oranı arasında anlamlı korelasyonlar saptandı (p<0,05) . Sonuç:Hastalarımızda KHC ve/veya Peg-INF-α'nın indüklediği tiroid bozuklukları yaygın olup, bunların çoğu otoimmundur ve kadınlarda daha sık idi. İlaveten, activin-A ve/veya follistatin TSHR-Abs oluşumu/agrevasyonunda etkili olabilir. Sonuçlarımızı doğrulamak ve KHC'daki tiroid hastalıkları oluşum mekanizmalarını açıklamak için ilave çalışmalara gereksinim vardır.Anahtar kelimeler: Kronik hepatit C, tiroid hastalığı, activin-A, follistatin, Suudi Arabistan ABSTRACTObjectives Chronic hepatitis C (CHC) and peg-interferon-α (Peg-INF-α) modulate serum activins and follistatin and are associated with thyroid disorders (TD).The aim of this study was to determine the frequency of CHC induced TD and to investigate the correlation of liver damage, serum activin-A and follistatin with the thyroid function parameters and thyroid autoantibodies. Methods:The study was cross-sectional and sera were obtained from 132 patients with CHC who were divided into 3 groups: 56 patients with no treatment, 30 after 24 weeks of Peg-INF-α and 46 at the end of 48 weeks Peg-INF-α. Thyroid stimulating hormone (TSH), free thyroxin (FT4), thyroid antibodies (Tabs), serum activin-A and follistatin levels were measured using ELISA.Results: Thyroid disorders were detected in 15% (n=20), more frequent in females (70%) and the majority were autoimmune thyroiditis (80%). TSH receptor antibodies (TSHRAbs) were significantly prevalent compared to the other antibodies (p<0.05) and significantly increased in the 24 weeks group (p<0.05). TSH and FT4 correlated significantly with liver enzymes (p<0.05). There was no significant difference in activin-A and follistatin values between thyroid disorder and euthyroids. However, significant correlations were found between TSHR-Abs ...
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