Non-rigid connector: The wand to allay the stresses on abutment

The use of rigid connectors in 5-unit fixed dental prosthesis with a pier abutment can result in failure of weaker retainer in the long run as the pier abutment acts as a fulcrum. Non-rigid connector placed on the distal aspect of pier seems to reduce potentially excess stress concentration on the pier abutment

Gene-Gene Interaction and Functional Impact of Polymorphisms on Innate Immune Genes in Controlling <em>Plasmodium falciparum</em> Blood Infection Level

<div><p>Genetic variations in toll-like receptors and cytokine genes of the innate immune pathways have been implicated in controlling parasite growth and the pathogenesis of <em>Plasmodium falciparum</em> mediated malaria. We previously published genetic association of <em>TLR4</em> non-synonymous and <em>TNF-α</em> promoter polymorphisms with <em>P.falciparum</em> blood infection level and here we extend the study considerably by (i) investigating genetic dependence of parasite-load on interleukin-12B polymorphisms, (ii) reconstructing gene-gene interactions among candidate <em>TLR</em>s and cytokine loci, (iii) exploring genetic and functional impact of epistatic models and (iv) providing mechanistic insights into functionality of disease-associated regulatory polymorphisms. Our data revealed that carriage of AA (P = 0.0001) and AC (P = 0.01) genotypes of <em>IL12B</em> 3′UTR polymorphism was associated with a significant increase of mean log-parasitemia relative to rare homozygous genotype CC. Presence of <em>IL12B+1188</em> polymorphism in five of six multifactor models reinforced its strong genetic impact on malaria phenotype. Elevation of genetic risk in two-component models compared to the corresponding single locus and reduction of <em>IL12B</em> (2.2 fold) and lymphotoxin-α (1.7 fold) expressions in patients'peripheral-blood-mononuclear-cells under <em>TLR4Thr399Ile</em> risk genotype background substantiated the role of Multifactor Dimensionality Reduction derived models. Marked reduction of promoter activity of <em>TNF-α</em> risk haplotype (C-C-G-G) compared to wild-type haplotype (T-C-G-G) with (84%) and without (78%) LPS stimulation and the loss of binding of transcription factors detected <em>in-silico</em> supported a causal role of <em>TNF-1031</em>. Significantly lower expression of <em>IL12B+1188</em> AA (5 fold) and AC (9 fold) genotypes compared to CC and under-representation (P = 0.0048) of allele A in transcripts of patients' PBMCs suggested an Allele-Expression-Imbalance. Allele (<em>A+1188C</em>) dependent differential stability (2 fold) of <em>IL12B</em>-transcripts upon actinomycin-D treatment and observed structural modulation (P = 0.013) of RNA-ensemble were the plausible explanations for AEI. In conclusion, our data provides functional support to the hypothesis that de-regulated receptor-cytokine axis of innate immune pathway influences blood infection level in <em>P. falciparum</em> malaria.</p> </div

<i>IL12B</i> 3′UTR based AEI analysis.

<p>(A) Representative electropherograms showing the peak heights of the sequence encompassing <i>IL12B+1188</i> locus in gDNA (lower panel) and cDNA (upper panel) extracted from the heterozygous (AC) patients' PBMC samples. The arrowhead indicates the +1188 site. Here A and C alleles are represented as green and blue peaks respectively. Difference in peak heights in sequences between cDNA and gDNA was evident. (B) Bar graph displays the A/C peak height ratios in individual heterozygous samples (N = 22). The dotted line indicates the baseline where the A/C ratio is one. (C) The departure of A/C peak height ratios from baseline in gDNA and cDNA were shown in the form of bar diagram. Each bar represents the mean height and corresponding standard deviation. The statistical difference of this distribution was measured by Sign test. P value has been indicated. (D) Dose-response and time-course assay of actinomycin-D treatment by real time PCR. (E) HepG2 cells (1×10⁵cells/ml) were treated with the optimum dose of actinomycin-D (5 µg/ml), harvested after 0 and 24 hours after treatment and <i>IL12B</i> mRNA levels were measured and corrected for <i>GAPDH</i> mRNA for both the wild-type and variant pSiCHECK2-3′UTR constructs. Statistical significance was measured by t-test. (*) indicates the P values to be statistically significant.</p

Association between <i>IL12B+1188</i> genotypes with blood infection level and <i>IL12B</i> gene-expression represented in Box plots.

<p>(A) Diagram represented the distribution of log-parasitemia across three genotypes: 11 (AA), 12 (AC) & 22 (CC) and (B) Diagram represented the comparison of log-parasitemia of minor homozygous genotype (CC) with AA and AC genotypic groups pooled. Statistical significance between pairwise comparisons was mentioned. (C) The ΔC_t distribution of <i>IL12B</i> gene expression across AA (N = 24), AC (N = 28) and CC (N = 12) genotypes and (D) comparison of gene expression between <i>IL12B+1188CC</i> genotype and with that of AA and AC individuals pooled together determined by quantitative real time PCR. Statistical significance was determined by the Mann Whitney U test. P values and fold changes obtained for each pairwise combination were appended in each plot. (*) indicates these differences to be statistically significant. The bottom, middle line, and top of each box correspond to the 25^th percentile, median, and the 75^th percentile, respectively. Bars extend to the lowest value and to the highest value of each group.</p