Alternative approaches to the therapeutic drug monitoring of prophylactic posaconazole in haematology patients.

OBJECTIVE: The intention of the study was to assess whether a unique daily specimen is adequate for prophylactic posaconazole TDM in haematology patients and if bioassay and HPLC produce similar results and could be equally used in clinical setting. METHOD: Serum specimens from thirty haematology patients were collected at the end of the first and second week of treatment, just before the morning dose, 2 and 6 to 8hours afterwards. Levels were measured by bioassay in 157 specimens and additionally by HPLC in 51 of them. RESULTS: Bioassay levels were correlated inter and intra daily, with no statistical difference between them, irrespective of the timing. The same was true for HPLC measurements. There was no statistical difference between bioassay (median: 1.60mg/L, interquartile range: 0.60-2.30) and HPLC levels (median: 1.16mg/L, interquartile range: 0.56-1.72), while they were significantly correlated. CONCLUSION: In clinically stable haematology patients, a random specimen on any day after steady state serum concentrations have been achieved is probably adequate in order to monitor posaconazole levels. In the case of monotherapy, a bioassay is an acceptable alternative to HPLC.

A Divalent PAMAM-Based Matrix Metalloproteinase/Carbonic Anhydrase Inhibitor for the Treatment of Dry Eye Syndrome.

Synthetic sulfonamide derivatives are a class of potent matrix metalloproteinase inhibitors (MMPI) that have potential for the treatment of diseases related to uncontrolled expression of these enzymes. The lack of selectivity of the large majority of such inhibitors, leading to the inhibition of MMPs in tissues other than the targeted one, has dramatically reduced the therapeutic interest in MMPIs. The recent development of efficient drug delivery systems that allow the transportation of a selected drug to its site of action has opened the way to new perspectives in the use of MMPIs. Here, a PAMAM-based divalent dendron with two sulfonamidic residues was synthesized. This nanomolar inhibitor binds to the catalytic domain of two MMPs as well as to the transmembrane human carbonic anhydrases (hCAs) XII, which is present in the eye and considered an antiglaucoma target. In the animal model of an experimental dry eye, no occurrence of dotted staining in eyes treated with our inhibitor was observed, indicating no symptoms of corneal desiccation.

Epidemiology of superficial mycoses in Northern Greece: a 4-year study.

BACKGROUND: Superficial mycoses are defined as the fungal infections of skin, hair or nails that are caused by dermatophytes, yeasts and non-dermatophytic moulds. Dermatophytes are the most frequently isolated fungi from specimens of patients with superficial mycoses. OBJECTIVE: Studying the possible alteration of the epidemiology of superficial mycoses in Northern Greece during the last two to three decades. MATERIALS AND METHODS: Data concerning the superficial mycoses from patients coming mainly from the region of Macedonia, Northern Greece, between January 2010 and January 2014 were recorded and analysed. They included specimens from 438 patients (146 M/292 F), within an age range of 2-85 years old. 503 samples were collected from skin (81, 16.1%), hair (18, 3.6%) and nails (fingernails 84, 16.7%, toenails 320, 63.6%) lesions. RESULTS: Of a total of 222 positive cultures, 50 were considered as yielding clinically non-significant isolates (saprophytes). Among the rest (172), dermatophytes were the most prevalent isolates (102, 59.3%), followed by yeasts (51, 29.7%) and non-dermatophytic moulds (19, 11%). Trichophyton rubrum (55, 53.9%), Trichophyton mentagrophytes (18, 17.6%) and Microsporum canis (23, 22.5%) were the most common isolates among dermatophytes (total = 102). Candida parapsilosis (26, 51%), and Candida albicans (10, 19.6%) among yeasts (total = 51) whereas Fusarium (6, 31.6%) and Acremonium species (3, 15.8%) among the non-dermatophytic moulds (total=19). CONCLUSION: Compared to previous studies from Northern Greece, the epidemiology of superficial mycoses in the specific geographic region seems not to have been altered the last two to three decades.

HLA-DQ relative risks for coeliac disease in European populations: a study of the European Genetics Cluster on Coeliac Disease.

Coeliac disease is an enteropathy due to an intolerance to gluten. The association between HLA-DQ genes and CD is well established. The majority of patients carry the HLA-DQ heterodimer encoded by DQA1*05/DQB1*02, either in cis or in trans. The remaining patients carry either part of the DQ heterodimer or DQA1*03-DQB1*0302. The aim of the study was to estimate the risks associated with different DQ genotypes in European populations. HLA information was available for 470 trio families from four countries: France (117), Italy (128), and Norway and Sweden (225). Five DQA1-DQB1 haplotypes were considered and control haplotype frequencies were estimated from the set of parental haplotypes not transmitted to the affected child. The possible genotypes were grouped into five genotype groups, based on the hierarchy of risk reported in the literature. A north-south gradient in the genotype group frequencies is observed in probands: homogeneity is strongly rejected between all country pairs. For each country, the relative risks associated with each genotype group were computed taking into account the control haplotype frequencies. Homogeneity of relative risks between countries was tested pairwise by maximum likelihood ratio statistics. The hypothesis of homogeneity of relative risks is rejected (P is approximately 10(-6)) for all country pairs. In conclusion, the gradient in the genotype group frequencies in probands is not only due to differences in haplotype frequencies but also due to differences in genotype relative risks in the studied populations; the relative risks associated with each DQ genotype group are different between northern and southern European countries; neither are they ordered in the same way.

HLA in coeliac disease: unravelling the complex genetics of a complex disorder.

Coeliac disease (gluten sensitive enteropathy) is a common, polygenic and multifactorial disorder that serves as a pioneering model for the study of inflammatory disease. A major environmental factor is known (ingested gluten from wheat), and there is unprecedented genetic and functional evidence pinpointing HLA-DQA1*05-DQB1*02 ( DQ2) and DQA1*03-DQB1*0302 ( DQ8) in disease predisposition. We discuss the current state of play in coeliac disease genetics, focussing particularly on the HLA complex. Emerging evidence suggests that additional HLA risk loci exert weak effects, independent of DQA1*05-DQB1*02, on the B8-DR3-DQ2 haplotype. There is also good evidence from linkage studies of disease gene(s) on chromosome 5q. We discuss the role and implications of linkage disequilibrium and haplotype blocks in complex disease gene mapping. We briefly address findings from studies of animal models for chronic inflammatory disease, and consider roles for both common genes associated with multiple inflammatory diseases, and genes unique to coeliac disease. The coeliac genetics research community has established a sound foundation for the identification of additional disease genes in the not-too-distant future. Functional studies will play a critical role, and coeliac disease has a promising future in this respect. Coeliac disease continues to function as a model disorder, facilitating the development and implementation of complex disease gene mapping strategies.

HLA in coeliac disease families: a novel test of risk modification by the 'other' haplotype when at least one DQA1*05-DQB1*02 haplotype is carried.

Predisposition to coeliac disease (CD) involves HLA genes. We investigated whether any haplotypes modify risk when carried trans to a known high-risk haplotype, DQA1*05-DQB1*02. Earlier attempts to rank levels of risk contributed by the 'other' haplotype were burdened by use of case-control populations; haplotype frequencies were estimated and homozygosity was only presumed. In contrast, exact haplotypes can be determined and allele transmission can be traced in families. A similar study in narcolepsy reported strata of different degrees of predisposition, attributable to the 'other' haplotype. A gene dosage effect similar to that described for DQB1*02 in CD, has also been reported in narcolepsy. We genotyped 439 simplex/multiplex trios for DQA1 and DQB1. We designed a new statistic to test risk modulation by the trans haplotype, even if the affected offspring was homozygous. We tested for significant deviation in transmission of the 'other' haplotype, i.e., modification of DQA1*05-DQB1*02 risk. We also addressed the proposed difference in risk, between DQA1*05-DQB1*02 homozygotes and DQA1*05-DQB1*02/DQA1*0201-DQB1*02 heterozygotes, reported in Southern Europe. We confirmed a DQB1*02 gene dosage effect. However, no haplotypes were found to modify risk when carried trans to DQA1*05-DQB1*02, except DQA1*05-DQB1*02 and DQA1*0201-DQB1*02 which were already known. We did not find credible evidence for a difference in risk conferred by DQA1*05-DQB1*02 and DQA1*0201-DQB1*02, when carried with DQA1*05-DQB1*02. The new test, which directly inspects haplotype transmissions rather than estimated haplotype frequencies, was used to demonstrate that the 'other' haplotype (except DQA1*05-DQB1*02 and DQA1*0201-DQB1*02) does not modify risk conferred by DQA1*05-DQB1*02. The test is applicable to other diseases.

Coeliac disease candidate genes: no association with functional polymorphisms in matrix metalloproteinase 1 and 3 gene promoters.

BACKGROUND: Coeliac disease is polygenic with a large genetic component. Matrix metalloproteinase-1 (MMP-1) and MMP-3 degrade extracellular matrix; expression levels are increased in the coeliac lesion where tissue damage is observed. Polymorphisms associated with elevated expression (MMP-3 -1171 allele 5A; MMP-1 -1607 2G), at 11q22.2, a region repeatedly showing evidence of linkage in coeliac disease, are associated with other chronic inflammatory disorders which may share a common molecular pathology. We tested for an association between these candidate gene polymorphisms and coeliac disease. METHODS: Two independent collections of 225 and 102 combined (Norwegian and Swedish) simplex families, and 160 independent healthy controls from the Norwegian Bone Marrow Donor Registry were used. Each individual was genotyped by PCR and fragment length analysis on an automated sequencer. The transmission/disequilibrium test was applied. Odds ratios were calculated employing probands or affected sibs where available, as cases versus independent controls. RESULTS: MMP-1 allele 2G did not show evidence of association in any tests undertaken. Neither did we find evidence for association of MMP-3 allele 5A, except among the combined family data: a non-significant tendency toward reduced risk was observed among males carrying MMP-3 allele 5A (40.2% transmission, Pc = 0.2). Further testing to clarify this observation did not reveal a significant association (odds ratio = 0.67 (95% confidence interval: 0.42-1.07), P = 0.08). CONCLUSIONS: We did not find significant evidence to support an association of MMP-3 allele 5A or MMP-1 allele 2G with coeliac disease in Norwegian and Swedish populations.

The IL12B gene does not confer susceptibility to coeliac disease.

Coeliac disease (CD) is a chronic inflammatory disorder where dietary gluten is not tolerated. In the lesion there are gluten reactive T cells predominantly secreting gamma-interferon. Both HLA and non-HLA genes contribute to CD susceptibility. Interleukin-12 (IL-12) regulates gamma-interferon production. The IL12B gene is located in a region (5q31.1-33.1) where there is evidence for linkage with CD. Allele 1 of an IL12B 3'UTR single-nucleotide polymorphism leads to increased expression of IL-12, and was recently implicated in susceptibility for type 1 diabetes (T1D). We found no evidence for association of allele 1 to CD by the transmission/disequilibrium test or case-control approach. No increased frequency was observed in patients belonging to families where the disease was linked to markers on chromosome 5q. Unlike T1D, allele 1 does not appear to confer susceptibility to CD.

Genes and environment in celiac disease.

Celiac disease is an intestinal disorder that develops as a result of interplay between genetic and environmental factors. HLA genes along with non-HLA genes predispose to the disease. Linkage studies have failed to identify chromosomal regions other than the HLA region which have major effects, indicating the existence of multiple non-HLA predisposing genes with modest effects. Association studies have shown that CTLA4 or a closely located gene is one of these genes. The primary HLA association in the majority of celiac disease patients is with DQ2 (DQA1*05/DQB1*02) and in the minority of patients with DQ8 (DQA1*0301/DQB1*0302). Gluten reactive CD4+ T cells can be isolated from small intestinal biopsies of celiac patients but not from controls. DQ2 or DQ8, but not other HLA molecules carried by patients, present peptides to these T cells. A number of distinct T cell gluten epitopes exist, most of them posttranslationally modified by deamidation. DQ2 and DQ8 bind the epitopes such that the glutamic acid residues created by deamidation are accommodated in pockets that have a preference for negatively charged side chains. There is evidence that deamidation in vivo is mediated by the enzyme tissue transglutaminase (tTG). Overall, the results point to control of the immune response to gluten by intestinal T cells restricted by the DQ2 or DQ8 molecules. This is likely to be a critical checkpoint for the development of celiac disease and could explain the dominant genetic role of HLA in this disorder. The products of the other predisposing genes may participate in pathway(s) that lead(s) to lesion formation. The minor genetic effects of the non-HLA genes could indicate a lack of critical checkpoints along these pathways, or that there are several pathways leading to the lesion formation.

Genome-wide linkage analysis of Scandinavian affected sib-pairs supports presence of susceptibility loci for celiac disease on chromosomes 5 and 11.

Celiac disease (CD) is a common chronic inflammatory disorder of the small intestine with a multifactorial aetiology. HLA is a well-known risk factor, but other genetic factors also influence disease susceptibility. To identify the genes involved in this disorder, we performed a genome-wide scan on 106 well-defined Swedish and Norwegian families with at least two affected siblings. We investigated familial segregation of 398 microsatellite markers, and utilised non-parametric linkage analysis. The strongest linkage with disease was found to the HLA locus (6p) (P<0.000006). There were eight regions besides HLA with a point wise P value below 0.05. Among these eight regions were 11q and 5q, both of which have been suggested in several linkage studies of independent celiac disease families. We also performed a stratification analysis of families according to their HLA genotypes. This resulted in significant differences on chromosome 2q. These results indicate that 11q, 5q and possibly also 2q are true susceptibility regions in CD.

Effect of bonded amalgam restorations on microleakage.

The purpose of this study was to evaluate quantitatively the effectiveness of adhesive dentin bonding systems in decreasing microleakage at the tooth-amalgam restoration interface. The results indicated that microleakage was significantly reduced when Amalgambond Plus or All-Bond 2 was used as liners in comparison to either Copalite varnish or no linear under amalgam restorations. No significant difference was found between the two dentin bonding systems at all time periods studied.

Esthetic treatment of anterior teeth. Technique choices.

The esthetic treatments of anterior teeth offer a variety of challenges. The dentist's artistic talents, understanding of the principles of esthetics and color are key points in planning and performing a successful treatment. The availability of different modalities of treatment should help provide the patient's needs and expectations. A thorough knowledge of the advantages and limitations of each treatment modality, and each clinical situation, should help in making appropriate treatment decisions.

A laboratory evaluation of the effect of two surface-wetting treatments on soft denture liners.

The wettability of acrylic and silicone soft denture-liners was improved by two methods of surface treatment. They consisted of formation of a thin film of silica from the application of silicon tetrachloride, and by cross-linking the surface molecules and bombarding with hydroxyl free radicals. Softness and water sorption were monitored throughout the study.