Regulatory Role of Promoter and 3′ UTR Variants of Vitamin D Receptor Gene on Cytokine Response in Pulmonary Tuberculosis

Vitamin D receptor (VDR) gene variants are shown to regulate immune response in tuberculosis. We studied the influence of VDR promoter (Cdx-2 and A1012G), 3' untranslated region (Apa I, Bsm I, and Taq I) and start codon (Fok I) polymorphisms on 1,25(OH)(2)D(3)-modulated IL-12p40, IFN-gamma, IL-10, and IL-5 response to live Mycobacterium tuberculosis and its culture filtrate antigen (CFA) in 60 normal healthy subjects and 51 pulmonary tuberculosis patients. In peripheral blood mononuclear cell cultures with CFA and 1,25(OH)(2)D(3), IL-12p40, and IFN-gamma levels were significantly decreased (p < 0.05) and IL-10 levels were significantly increased (p < 0.05) in patients with GG genotype. The extended genotype bbaaTT (baT haplotype) was associated with decreased IL-12p40 and IFN-gamma levels and significantly increased IL-10 levels (p < 0.05). The Cdx-2 GG genotype and baT haplotype are associated with a suppressed Th1 and increased IL-10 response, which suggests that 1,25(OH)(2)D(3) probably through the VDR polymorphic variants augments the anti-inflammatory response at the site of M. tuberculosis infection.     

Genetic Variation in the 3′ Untranslated Region of the Neurofibromatosis 1 Gene: Application to Unequal Allelic Expression

Neurofibromatosis type 1 (NF1) is a common genetic disorder caused by inactivation of neurofibromin, a protein capable of modulating signal transduction by activating Ras-GTPase activity. We have used cDNA cloning and Northern blot analysis to confirm the NF1 gene produces alternatively polyadenylated mRNAs with 3 untranslated regions (3UTR) that show striking evolutionary conservation. Scanning of the 3UTRs for genetic variation revealed three common sequence polymorphisms (>30% heterozygosity), one less informative polymorphism (5% heterozygosity) and one rare variant (1/144 chromosomes). These differences were used to examine relative levels of expression of normal and mutant NF1 alleles in lymphoblast cell lines and in one case, autopsy tissue, from patients with NF1. Unequal allelic expression (up to 4-fold) was observed in a subset of both sporadic and familial NF1 cases. Where linkage phase could be determined, the allele segregating with the disorder displayed a relative reduction in expression. However, the magnitude of this effect was variable suggesting the operation of additional, non-genetic factors in determining the degree of relative expression of the mutant allele.     

Association of Crohn’s Disease with Aryl Hydrocarbon Receptor Gene Polymorphisms in Patients from Southeast China

ABSTRACT

Aims The aryl hydrocarbon receptor (AhR) plays a pivotal role in regulating the innate and the acquired immune systems. The present study aimed to investigate the association of Crohn’s disease (CD) with AhR polymorphisms in a cohort of patients from Southeast China.

Methods An improved multiple ligase detection reaction technique was applied to examine the polymorphisms of rs2158041, rs2066853, and rs10249788 in 310 patients with CD and 573 controls.

Results Compared to the controls, the variant allele (T) and genotype (CT+TT) of rs2158041 were less frequent in patients with CD (both p < 0.05). Similar conclusions were drawn from patients with ileal CD and with stricture CD as compared to the controls (all p < 0.0083). However, no significant differences were observed in allele and genotype frequencies of rs2066853 and rs10249788 between patients with CD and the controls (all p > 0.05). Although rs2158041 and rs10249788 were in complete linkage disequilibrium with rs2066853, respectively, only the frequency of haplotype (TG) formed by rs2158041 and rs2066853 was significantly lower in patients with CD than that in the controls (p < 0.05).

Conclusions AhR (rs2158041) might be a susceptible locus for CD, especially for the two subtypes: ileal CD and stricture CD.     

Gender-related Distribution of the Interleukin-1β and Interleukin-1 Receptor Antagonist Gene Polymorphisms in Patients with End-stage Liver Disease

Interleukin-1β (IL-1β) genetic polymorphisms and IL-1 receptor antagonist (IL1RN) variable number tandem repeat (VNTR) seem to be related with the occurrence of chronic diseases. This study aimed to verify whether IL-1β -511>C/T, -31>T/C, +3953>C/T and IL1RN VNTR were associated to the development of liver cirrhosis. Two hundred forty cirrhotic patients were involved in the study. A significant trend was detected, for increasing cirrhosis frequencies, grouping the patients as follows: females and males carrying neither the IL-1β (-511 -31) T-C/T-C or T-C/(T-T or C-C) diplotypes nor any IL1RN A2 allele (138/292), males carrying either the IL-1β T-C/T-C or T-C/(T-T or C-C) diplotypes or at least one IL1RN A2 allele (74/147) and males carrying either the IL-1β T-C/T-C or T-C/(T-T or C-C) diplotypes and at least one IL1RN A2 allele (28/37) (p?<?0.01). IL-1β polymorphisms are associated with the occurrence of end stage liver disease. IL-1β inflammatory activity appears more pronounced in males.     

Genetic diversity of 3′ region of glycoprotein D gene of bovine herpesvirus 1 and 5

Bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5) are closely related alphaherpesviruses of cattle. While BoHV-1 is mainly associated with respiratory/genital disease and rarely associated with neurological disease, BoHV-5 is the primary agent of meningoencephalitis in cattle. The envelope glycoprotein D of alphaherpesviruses (BoHV-1/gD1 and BoHV-5/gD5) is involved in the early steps of virus infection and may influence virus tropism and neuropathogenesis. This study performed a sequence analysis of the 3′ region of gD gene (gD3′) of BoHV-1 isolates recovered from respiratory/genital disease (n = 6 and reference strain Cooper) or from neurological disease (n = 7); and from seven typical neurological BoHV-5 isolates. After PCR amplification, nucleotide (nt) sequencing, and aminoacid (aa) sequence prediction; gD3′ sequences were compared, identity levels were calculated, and selective pressure was analyzed. The phylogenetic reconstruction based on nt and aa sequences allowed for a clear differentiation of BoHV-1 (n = 14) and BoHV-5 (n = 7) clusters. The seven BoHV-1 isolates from neurological disease are grouped within the BoHV-1 branch. A consistent alignment of 346 nt revealed a high similarity within each viral species (gD1 = 98.3 % nt and aa; gD5 = 97.8 % nt and 85.8 % aa) and an expected lower similarity between gD1 and gD5 (73.7 and 64.1 %, nt and aa, respectively). The analysis of molecular evolution revealed an average negative selection at gD3′. Thus, the phylogeny and similarity levels allowed for differentiation of BoHV-1 and BoHV-5 species, but not further division in subspecies. Sequence analysis did not allow for the identification of genetic differences in gD3′ potentially associated with the respective clinical/pathological phenotypes, yet revealed a lower level of gD3′ conservation than previously reported.     

Deglycosylation of Serum Vitamin D3-Binding Protein by α-N-Acetylgalactosaminidase Detected in the Plasma of Patients with Systemic Lupus Erythematosus

A serum glycoprotein, Gc protein (vitamin D₃-binding protein), can be converted by β-galactosidase of B cells and sialidase of T cells to a potent macrophage-activating factor (MAF), a protein withN-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is the precursor for MAF. Treatment of Gc protein with immobilized β-galactosidase and sialidase generates a remarkably high titered macrophage-activating factor (GcMAF). When peripheral blood monocytes/macrophages (designated macrophages) of 33 systemic lupus erythematosus patients were incubated with GcMAF (100 pg/ml), the macrophages of all patients were activated as determined by superoxide generation. However, the precursor activity of patient plasma Gc protein was lost or reduced in these patients. Loss of the precursor activity was the result of deglycosylation of plasma Gc protein by α-N-acetylgalactosaminidase activity found in the patient plasma. Levels of plasma α-N-acetylgalactosaminidase activity in individual patients had an inverse correlation with the MAF precursor activity of their plasma Gc protein. Deglycosylated Gc protein cannot be converted to macrophage-activating factor. The resulting defect in macrophage activation may lead to an inability to clear pathogenic immune complexes. Thus, elevated plasma α-N-acetylgalactosaminidase activity resulting in the loss of MAF precursor activity and reduced macrophage activity may play a role in the pathogenesis of systemic lupus erythematosus.     

Evaluation of Two Hepatitis C Virus Genotyping Assays Based on the 5′ Untranslated Region (UTR): the Limitations of 5′ UTR-Based Assays and the Need for a Supplementary Sequencing-Based Approach

We evaluated two genotyping methodologies that characterize the 5′ untranslated region (5′ UTR) of the hepatitis C virus (HCV) genome. The limitations of these genotype assays need to be thoroughly evaluated, and sequencing-based approaches may be needed to complement these methods in clinical settings.     

Effects of μ-Opioid Receptor Gene Polymorphism on Postoperative Nausea and Vomiting in Patients Undergoing General Anesthesia with Remifentanil: Double Blinded Randomized Trial

Association between postoperative nausea and vomiting (PONV) and µ-opioid receptor A118G single nucleotide polymorphism (SNP) is undefined and might underlie inconsistent results of studies on PONV occurrence in patients undergoing general anesthesia with the opioid, remifentanil. Four hundred and sixteen Korean women undergoing breast surgery with general anesthesia were randomized to receive remifentanil 10 ng/mL (plasma-site, Minto model) using a target-controlled infusion device and either propofol for total intravenous anesthesia (T group) or sevoflurane for inhalation anesthesia (I group) with bispectral index values maintained between 40 and 60. Blood specimens were collected after anesthesia induction for A118G SNP analysis. PONV and postoperative pain were evaluated. A118G SNP type distribution among Korean female adults studied was AG (n=195)>AA (n=158)>GG (n=63). Regardless of anesthetic technique, patients with GG types had lower PONV scale on arrival at postoperative care unit (PACU) (P=0.002), while T group showed lower PONV scale than I group up to 6 hr after PACU discharge in AA and AG types. No differences were apparent for postoperative pain among opioid receptor polymorphism. PONV occurrence differs according to opioid receptor polymorphism and anesthetic technique in patients undergoing general anesthesia with remifentanil.

Graphical Abstract

相似文献   

Fascin Regulates TLR4/PKC-mediated Translational Activation Through miR-155 and miR-125b,which Targets the 3′ Untranslated Region of TNF-α mRNA

Fascin is a well-known cytoskeletal regulatory protein that, as a substrate of protein kinase C (PKC), is involved in PKC-mediated translational regulation of TNF-α in macrophages stimulated with lipopolysaccharide (LPS). The regulatory effects of fascin targeted the 3′-untraslated region (UTR) of the TNF-α mRNA, and suppression of PKC activity or fascin expression resulted in specific blockage of the LPS-induced translational activation of the mRNA. In an effort to identify the molecular mechanism of this fascin-mediated translational regulation, the expression levels of micro-RNA (miRNA) after stimulation of the toll-like receptor 4 (TLR4) signaling pathways were analyzed in cells with down-regulation of fascin. The LPS-induced translation of TNF-α is known to be regulated by miR-155 and miR-125b, which have positive and negative effects, respectively. Interestingly, suppression of fascin expression reversed LPS-induced down-regulation of miR-125b and abolished the LPS-induced increase in miR-155. Furthermore, introduction of miR-155 precursor, blocking of miR-125b activity, or introduction of a mutation into the miR-125b binding site of the TNF-α 3′-UTR restored translational activation in cells with suppressed fascin expression. These data indicate that fascin regulates translation through miR-155 and miR-125b, which target 3′ UTR in TNF-α mRNA.     

Phylogenetic Analysis of 5′-UTR and P1 Protein of Indian Common Strain of Potato Virus Y Reveals its Possible Introduction in India

The 5' untranslated region (UTR) and P1 region of the Indian strain of potato virus Y ordinary strain (PVYO) was cloned and sequenced for the first time. Database searches and multiple sequence alignment showed the highest sequence similarity with the PVYO strains of European origin. Based on the phylogenetic analysis and multiple sequence alignment, the possible evolution of PVYN from PVYO is predicted. PVYO strains from China and India were perhaps introduced into these countries from a similar geographical location. All major PVY strains available in the database can be classified into two major subgroups of North American and European origin. The Chinese and Indian PVYO strains fall within the European union subgroup suggesting a long association since potato was introduced from Europe into these countries by two separate independent events. The possible function of P1 protein in plant virus replication is suggested due to in-silico prediction of nuclear localization signal (NLS) and other phosphorylation regulatory domains at the vicinity of the NLS.     

Systematic Review and Meta–Analysis of the Diagnostic Accuracy of Fibrosis Marker Panels in Patients with HIV/Hepatitis C Coinfection

Abstract

Background: Accurately staging hepatitis C virus (HCV)–related fibrosis is crucial for treatment decisions and prognostication. Our objective was to systematically review studies describing the accuracy of serum marker panels for predicting fibrosis in HIV/HCV–coinfected patients. Method: Studies comparing serum marker panels with biopsy in HIV/HCV-coinfected patients were identified. Random effects meta-analyses and areas under summary receiver operating characteristics curves (AUC) examined test accuracy for detecting significant fibrosis (F2–4) and cirrhosis. Heterogeneity was explored using meta-regression. Results: Five studies (n = 574) including four fibrosis measures (APRI [n = 4 studies], Forns’ [n = 2], FibroTest [n = 1], SHASTA [n = 1]) met the inclusion criteria. The prevalence of significant fibrosis and cirrhosis were 51% and 16%, respectively. For the prediction of significant fibrosis, the summary AUC was 0.82 (95% CI 0.78–86) and diagnostic odds ratio was 7.8 (5.1–11.9). For cirrhosis, these figures were 0.83 (0.69–0.97) and 11.0 (4.6–26.2), respectively. Meta–regression including study factors (methodological quality and biopsy adequacy), patient characteristics (age, gender, CD4 count), and fibrosis measure failed to identify important predictors of accuracy. Conclusion: Available fibrosis marker panels have acceptable performance for identifying significant fibrosis and cirrhosis in HIV/HCV–coinfected patients but are not yet adequate to replace liver biopsy. Additional studies are necessary to identify the optimal measure.     

Analysis of Treatment Costs for HIV RNA Reductions and CD4 Increases for Darunavir Versus Other Antiretrovirals in Treatment-Experienced,HIV–Infected Patients

Abstract

Background: For patients with preexisting HIV drug resistance, a wide range of antiretrovirals are used, with differences in efficacy and cost. The additional cost per incremental 25 cell rise in CD4 count, or 0.5 log reduction in HIV RNA, was calculated for 8 antiretrovirals using pivotal clinical trials data. Method: For approved antiretrovirals in HIV therapy–experienced patients, 24–week efficacy (benefit over control in HIV RNA and CD4 count) was extracted from pivotal trials in published reports and compared with the additional treatment cost versus the control arm of each trial (2006 US wholesale acquisition costs). Treatment costs in the POWER trials were calculated directly from the treatment use database. Results: Data were available from 11 clinical trials in more than 4,000 antiretroviral treatment-experienced patients: Gilead 907 (TDF vs. placebo), TORO1/2 (T-20/OBR vs. OBR), RESIST–1/2 (TPV/r vs. control PI), BMS-045 (ATV/r vs. LPV/r), CONTEXT (fAPV/r vs. LPV/r), CAESAR (3TC vs. placebo), CNA3002 (ABC vs. placebo), and POWER 1/2 (DRV/r vs. control PI). Additional cost per 0.5 log reduction in HIV RNA was $152 for ritonavir–boosted darunavir (DRV/r), $4,453 for lamivudine (3TC), $4,274 for abacavir (ABC), $4,641 for tenofovir (TDF), and $13,217 for enfuvirtide (T-20). Cost per 25 cell rise in CD4 ranged from $132 for darunavir/r to $16,464 for T-20. Conclusion: There is a wide range of costs associated with efficacy improvements across the classes of antiretrovirals used for antiretroviral treatment–experienced patients. This analysis does not account for differences in toxicity, use of concomitant medications, or long-term adherence, which could also influence value assessments.     

Changing use of traditional healthcare amongst those dying of HIV related disease and TB in rural South Africa from 2003 – 2011: a retrospective cohort study

Background

In 2011 there were 5.5 million HIV infected people in South Africa and 71% of those requiring antiretroviral therapy (ART) received it. The effective integration of traditional medical practitioners and biomedical providers in HIV prevention and care has been demonstrated. However concerns remain that the use of traditional treatments for HIV-related disease may lead to pharmacokinetic interactions between herbal remedies and ART drugs and delay ART initiation. Here we analyse the changing prevalence and determinants of traditional healthcare use amongst those dying of HIV-related disease, pulmonary tuberculosis and other causes in a rural South African community between 2003 and 2011. ART was made available in this area in the latter part of this period.

Methods

Data was collected during household visits and verbal autopsy interviews. InterVA-4 was used to assign causes of death. Spatial analyses of the distribution of traditional healthcare use were performed. Logistic regression models were developed to test associations of determinants with traditional healthcare use.

Results

There were 5929 deaths in the study population of which 47.7% were caused by HIV-related disease or pulmonary tuberculosis (HIV/AIDS and TB). Traditional healthcare use declined for all deaths, with higher levels throughout for those dying of HIV/AIDS and TB than for those dying of other causes. In 2003-2005, sole use of biomedical treatment was reported for 18.2% of HIV/AIDS and TB deaths and 27.2% of other deaths, by 2008–2011 the figures were 49.9% and 45.3% respectively. In bivariate analyses, higher traditional healthcare use was associated with Mozambican origin, lower education levels, death in 2003–2005 compared to the later time periods, longer illness duration and moderate increases in prior household mortality. In the multivariate model only country of origin, time period and illness duration remained associated.

Conclusions

There were large decreases in reported traditional healthcare use and increases in the sole use of biomedical treatment amongst those dying of HIV/AIDS and TB. No associations between socio-economic position, age or gender and the likelihood of traditional healthcare use were seen. Further qualitative and quantitative studies are needed to assess whether these figures reflect trends in healthcare use amongst the entire population and the reasons for the temporal changes identified.

     

Induction of an ATP-polymerizing enzyme in TMV-infected tobacco and its homology to the human 2′–5′ a synthetase

Several reports have indicated that tobacco carries an enzyme (APE) that, in the presence of poly (rI):(rC), polymerizes ATP to oligoadenylates. This paper demonstrates that the tobacco APE system comprises several proteins (estimated sizes: 32, 42, 67, and 84±10% kD). Only one of these proteins (the 67-kD form) binds to poly (rI):(rC). This APE form has been purified by affinity chromatography on a synthetic ds-RNA column. Four tobacco proteins, including the purified one, crossreact with antibodies against the human enzyme, 2–5 A synthetase. The ATP-binding capacity of some of these proteins has also been demonstrated. The amount of plant oligoadenylates obtained by polymerizing ATP with the purified APE form allows, for the first time, their direct analysis by TLC. The TLC analysis indicated that the oligomer produced by APE is not identical to the 2–5 oligoadenylate.     

Vitamin D3 and Calcipotriol Enhance the Secretion of Transforming Growth Factor-β1 and -β2 in Cultured Murine Keratinocytes

Abstract

Vitamin D₃ and its analogue calcipotriol (MC 903) inhibit the proliferation of cultured keratinocytes and induce their differentiation. Since TGFβs are very potent inhibitors of keratinocyte growth we studied the effects of vitamin D₃ and calcipotriol on the secretion of TGFβ in cultured murine keratinocytes. Vitamin D, and calcipotriol (10^?6 - 10^?9 M) inhibited the DNA-synthesis of mouse keratinocytes by 50–80% in a time and dose-dependent manner as measured by [³H]-thymidine incorporation. Analysis of the conditioned medium of the keratinocytes indicated that the cells secreted into their medium activity that inhibited the growth of indicator Mv1Lu mink lung epithelial cells. Neutralizing antibodies against TGFβ1 and TGFβ2 decreased, and when used together, prevented the observed growth inhibition of the indicator cells. Heat treatment of the conditioned medium, which activates latent forms of TGFβ, revealed higher levels of growth inhibitory activity in the medium from vitamin D₃ and calcipotriol treated than from control cultures indicating that a fraction of TGFβ was in a latent form. Active TGFβ was, however, detected considerably more in vitamin D₃ and calcipotriol treated cultures than in control cultures. Immunoblotting analysis of the medium revealed enhanced secretion of TGFβ protein. These results indicate that enhanced TGFβ1 and TGFβ2 secretion and activity is associated with vitamin D₃-mediated growth inhibition of cultured keratinocytes.

This work was presented in part at the Keystone Symposium “Negative Growth Control”, Keystone, CO, Jan. 26-Feb. 2, 1992 (Koli and Keski-Oja 1992).