Tissue transglutaminase (tTG) autoantibodies decline after gluten-free diet in patients with coeliac disease. We tested the hypothesis that gluten-free diet-induced change in tTG autoantibody levels affects subsets of peripheral blood lymphocytes. Peripheral blood was obtained from 20 children with biopsy-proven active coeliac disease. Gluten-free diet was initiated and the children examined again after three and six months. tTG autoantibodies were measured in radioligand binding assays and lymphocyte subsets by flow cytometry. IgA-tTG levels at diagnosis, 2204 U/ml (median, range 113-24990), were reduced over six months to 76 U/ml (median, range 1-1261) (P < 0.001). At six months, 12/20 (60%) children had reduced their IgA-tTG levels to < 100 U/ml and these children showed a decrease in B cells (mean change -3.8%, P = 0.014), CD4+ T cells (mean -4.32%, P = 0.011) and CD4+ T cells expressing CD25high (mean change -0.62%, P = 0.036). In contrast, the CD4+CD25(high)CCR4+ T cell population increased during the same period (mean change 11.5%, P = 0.0036). The decline in IgA-tTG levels correlated to the decrease in B cells (r = 0.56, P = 0.01), CD4+ T cells (r = 0.66, P = 0.004) as well as CD4+CD25high T cells (r = 0.59, P = 0.01). A negative correlation was found between the decline in IgA-tTG and CD4+CD25high T cells expressing CD45RO (r = -0.49, P = 0.03) and CCR4 (r = -0.54, P = 0.01). This is the first observational study on the effect of gluten-free diet on concurrent changes of tTG autoantibodies and specific peripheral blood lymphocyte subsets. Our data suggest that flow cytometry may be a useful complement to tTG autoantibodies when studying the effects of gluten-free diet in children with coeliac disease.

Celiac disease (CD) is an (auto)immunologically mediated intestinal intolerance against proteins from wheat (gluten) and related cereal proteins. Tissue transglutaminase (tTG) has been identified as the autoantigen in CD. Although ultimate diagnosis is based on histological analysis of small intestinal mucosa obtained via tissue biopsy, assessment of autoantibodies can provide substantial help in the evaluation of CD. Gliadin antibodies are directed against the native disease-provoking cereal proteins. Despite their initial usefulness, these antibodies have lost diagnostic importance due to their poor specificity and sensitivity as CD markers. Recently, it was found, however, that gliadin antibodies from sera of patients with active CD preferentially recognized deamidated gliadin peptides. The use of deamidated gliadin peptides in immunoassays has significantly improved the usefulness of gliadin antibodies in diagnosis of CD to that observed with autoantibody assay methods (endomysium antibodies, antibodies against tTG). The antibody epitopes (B-cell epitopes) reflect substrate specificity of tTG and resemble peptide sequences known to be strongly T-cell stimulatory (T-cell epitopes) in CD. The assay applying deamidated gliadin peptides measures a new species of antibodies, which is different from conventional gliadin antibodies as well as from autoantibodies and will likely provide new information on pathophysiological mechanisms of CD.

BACKGROUND

Celiac disease (CD) is induced by wheat gluten and related prolamines. Its prevalence may be underestimated in many geographic regions and populations, and has recently increased in several countries. In 1998 and 1999, a random sample of Estonian schoolchildren was screened with IgA-type tissue transglutaminase antibodies (IgA-tTG) for CD. The results revealed a CD prevalence of 0.34%, which is lower compared with many other European countries.

OBJECTIVE

We rescreened the same population for CD using IgA-tTG after a 10-year interval.

METHODS

A total of 891 patients from the initial sample were rescreened using the IgA-tTG assay for a participation rate of 76.8% (median age, 24.3 years). As in the initial study, the IgA-tTG results were evaluated by ImmunoCAP EliA Celikey using an IgG-tTG and deamidated gliadin antibody assay for IgA-deficient cases.

RESULTS

No new cases of CD were found in this follow-up study. Of note, 75% of patients with initial IgA-tTG-positive results and biopsy-proven CD remained seropositive. One patient with a negative seroconversion at the time of rescreening followed a strict gluten-free diet during the follow-up years.

CONCLUSIONS

In a 10-year follow-up period, no new cases of CD were found in this Estonian population of school-children and young adults. Therefore, we presume no increase in CD during the last decade among this age group in Estonia. Additional studies are needed to determine whether similar results would be obtained in other age groups, because of differences in the CD prevalence between Estonian and other European populations.

A randomized, double-blind, placebo-controlled trial was conducted to (1) evaluate efficacy and safety of transdermal testosterone gel (AndroGel) for hypogonadal men in Taiwan, and (2) observe improvements in sexual function through international index of erectile function (IIEF) scores. Eligible hypogonadal men were randomized to receive 50 mg/day transdermal testosterone gel (TTG) or placebo for 3 months. Primary end point was change from baseline in total testosterone (TT) and free testosterone (FT). Secondary end points were change from baseline in serum hormone levels (such as dihydrotestosterone (DHT), estradiol (E2), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and sex-hormone-binding globulin (SHBG)) and changes in IIEF scores. Safety evaluations included adverse events (AEs) and skin irritation assessment. Compared with baseline, the TTG group (n=20) had statistically significant increases in mean TT levels at month 1 (P=0.024) and month 2 (P=0.025), but no significant changes at month 3. TT levels in the placebo group (n=18) showed no statistically significant change at any visit. Changes in FT levels paralleled changes in TT levels in both groups. TTG group IIEF scores were significantly increased at month 3 (P=0.01), compared with a decline in placebo scores. No drug-related AEs occurred in the TTG group; the placebo group had 2 AEs (mild skin rash). In conclusion, TTG effectively restores serum TT and FT levels to a normal physiological range for hypogonadal men in Taiwan and improves sexual function.

Celiac disease (CD) is a chronic inflammatory enteropathy of the small bowel resulting from a local TH1-mediated reaction to wheat gliadins and barley, rye and oat prolamins with the development of auto-antibodies to transglutaminases. As well as for other chronic inflammatory diseases, genetic background and environmental factors participate to pathogenesis. An increased traffic of CD34+ hemopoietic precursor and stem cells (HPC) has been reported in peripheral blood (PB) of subjects with allergic diseases that share in their pathogenesis immuno-mediated reactions, genetic and environmental factors. The aim of the present work was to investigate the CD34+ cell traffic and H2/H1 polarization of lymphoid T-cell lineage, in the peripheral blood of subjects with CD, by means of flow-cytometric techniques. Group A of control was of 20 healthy subjects, aged 5 to 58 years. Study population (Group B) was of twenty-eight patients, all females aged 13 to 70, receiving firstly a CD diagnosis at the SS Annunziata Hospital Digestive Physiopathology Out-standings' by means of clinical, serologic and small intestinal biopsy findings. Peripheral CD34+ HPCs were significantly increased in Group B (median value 0.16) when compared with Group A (median value 0.03) (p 0.0001) but did not correlate either with anti-transglutaminase (tTG) antibody levels (IgA: p 0.226; IgG: p 0.810) or with histological damage severity (p 0.41) that, on the contrary, was significantly related with anti-tTG IgA antibodies (p 0.027). Celiac circulating CD3+CD4+ lymphocytes expressed a chemokine-receptor pattern Th2-skewed in all but three patients investigated. Concluding, the CD34+ HPC highly increased peripheral traffic observed in celiac disease appears more related to a basic and emerging as common defect shared by chronic inflammatory diseases than to the gliadin-specific Th1 local reactions. Data are consistent with a potential NFkappaB deficiency and consequent prevalence of apoptotic versus survival programs leading to excessive cell-death; to replace lost cells a supplementary bone-marrow derived precursors supply, further to that physiologically provided by the gut stem cell "niches" that are cryptopatches, could be required.

OBJECTIVE

The aim of this article is to determine the prevalence of celiac disease (CD), Helicobacter pylori (H. pylori) and gastroesophageal reflux (GER) in patients with resistant iron deficiency anemia (IDA).

METHODS

The study included 25 patients <18 years of age with refractory IDA (not responding to iron therapy for 3 months in a dose of 6 mg elemental iron/kg/day).

METHODS

All patients included in the study were subjected to careful history taking and thorough clinical examination. Blood sample was taken for analysis of antibodies for CD including: antigliadin antibody (AGA), antiendomysial antibody (EMA), antireticulin antibody (ARA) and antitissue Transglutaminase (tTg) IgG antibody. Anti-H. pylori IgG antibodies and a (13)C-urea breath test (UBT) was done to all patients to diagnose H. pylori. Upper gastrointestinal tract endoscopy was done for all patients to evaluate for the presence of some etiologies of intractable anemia as chronic blood loss. These included: CD, H. pylori infection and GER. The upper gastrointestinal tract endoscopy was also done to evaluate the presence of bleeding spots, ulcers or angiomatous malformations. In addition, gastric antral biopsies were taken for diagnosis of H. pylori infection by the following tests: rapid urease test, histopathological examination and culture.

RESULTS

CD was positive in 11 out of 25 patients (44%), H. pylori infection in 12 out of 25 patients (48%), while GER was diagnosed in 11 out of 25 patients (44%). Patients with CD had age of presentation < or =2 years in two patients (18.2%) while the remaining nine patients (81.8%) had age of presentation >2 years and it was statistically significant (p = 0.05*). Also patients with H. pylori had age of presentation < or =4 years in five patients (41.7%) and the remaining seven patients (81.8%) had age of presentation >4 years and it was statistically significant (p = 0.03*). Logistic regression analysis demonstrated that the risk factors for severity of anemia were age of patients and duration of anemia. On the other hand, other parameters have no significant influence on the severity of anemia. Also risk factors of short stature were age of presentation of anemia, degree of anemia and H. pylori infection. AGA had the highest sensitivity (100%) followed by antiendomysium antibody (81.8%) while the tTG antibody had the highest specificity (85.7%) for diagnosis of CD. UBT and histopathology had the highest sensitivity (100%) for diagnosis of H. pylori while rapid urease test, culture, H. pylori stool antigen and anti-H. pylori IgG antibody had the highest specificity (100%). In conclusion, refractory IDA may be due to clinically unapparent H. pylori gastritis and CD. CD is one of the most common causes of intestinal malabsorption during childhood which leads to impairment of iron absorption. Apart from offering them gluten-free diet rich in iron, early detection and treatment of IDA and prophylactic iron and folic acid supplementation will go a long way to optimize their mental and psychological functions. Eradication of H. pylori infection with concomitant iron therapy should correct the anemia.

Partial mitochondrial (mt) genome sequence (10,486bp) from the parasitic nematode Toxocara vitulorum was determined and its organization and structure compared with those of T. cati, T. canis and T. malaysiensis. The obtained mt genome sequence of T. vitulorum contains 10 protein-coding genes (cytochrome c oxidase subunits 1-3, Nicotinamide adenine dinucleotide dehydrogenase subunits 1-5, ATP synthase subunit 6 and cytochrome b), 14 transfer RNA genes and the large ribosomal RNA gene (rrnL), non-coding regions. ORF encoding for ATPase subunit 8 is not found in this partial mtDNA sequence. Five translation initiation codons were inferred, ATT, ATG, GTG, GTT and TTG. Most of the genes used TAG or TAA as a stop codon and two genes ended with a T. The gene arrangement and composition of the T. vitulorum mt genome is very similar to that of other Toxocara species mitochondrial genomes sequenced thus far. All genes are transcribed in the same direction, as other Toxocara species. This genome has a high A+T content (67.5%) and low G+C content (32.5%). Phylogenetic reconstruction based on aligned nucleotide sequences of seven taxa provided strong support that Toxocara vitulorum is more closely related to T. malaysiensis than to T. canis and T. cati.

BACKGROUND

Gastrointestinal (GI)-related autoantibodies (Abs) are rarely evaluated in autoimmune diseases (AID) other than inflammatory bowel disease, autoimmune hepatitis and celiac disease. Our aim was to determine the prevalence of these antibodies in a wide spectrum of AID.

METHODS

We examined 923 serum samples representing 18 AID and compared them with 338 samples from healthy subjects. We used the BioPlex 2200-immunoassay (Bio-Rad, USA) to test samples for the presence of IgA and IgG directed at gliadin (AGA), tissue-transglutaminase (tTG), and Saccharomyces cerevisiae (ASCA).

RESULTS

Prevalence of IgA AGA was significantly higher in antiphospholipid syndrome (APS) (7.1 %, P=0.012) and in pemphigus vulgaris (25%, P =0.008) patients, as compared with healthy controls. Presence of IgG-AGA was more common among Crohn's disease (20.5%, P = 0.023) and rheumatoid arthritis (6.5%, P=0.027) patients. IgG anti tTG were frequently observed in APS (6.1%, P=0.012), in giant cell arteritis (11.5%, P=0.013) and in ulcerative colitis (11.1%, P=0.018) patients, and as expected, higher prevalence of ASCA (IgA 19.3% and IgG 27.7%) was found in Crohn's disease. IgG ASCA were also found in systemic lupus erythematosus (SLE) (4.5%, P=0.01), in Graves' disease (5.7%, P=0.018), in cryoglobulinemia (7.1%, P=0.006), and in patients with vasculitides (6.5%, P=0.002). In contrast, lower prevalence of IgG type AGA was found in SLE (P=0.034), cryoglobulinemia (P=0.019) and vasculitides (P=0.013) patients.

CONCLUSIONS

Our findings suggest an association between GI-related- Abs and a wide spectrum of AID. The clinical implication of these findings is yet to be determined.

Comparison of the cya loci (cya codes for adenylyl cyclase (AC)) from a variety of phylogenetically divergent facultative anaerobic Gram-negative bacteria reveals conserved sequence features. The entire locus structure in enterobacteria is preserved, including two major promoters (a conserved cya strong promoter, P2, and a divergent promoter for a heme biosynthetic operon, hemCD) present in the upstream region of the cya gene. The region between hemC and cya is much longer in Proteus mirabilis than in other enterobacteria, and lacks the P1 upstream cya promoter. In Aeromonas hydrophila the cya promoter (the strong P2 promoter in E coli) is preserved, including a putative GATC methylation site situated immediately downstream from the -10 box. Each cya frame analyzed uses TTG as the translation start codon and is preceded by an unusual ribosome binding site. This suggests that a lower translation efficiency of the cya transcript could be the result of some selection pressure. This has been substantiated by in vitro mutagenesis and by selection of up mutations which all map at the cya ribosome binding site. In enterobacteria the cyaY frame is the only conserved reading frame downstream of cya, with the orientation opposite to that of cya. This organization is not preserved in Aeromonas. Experiments involving fusions with the lacZ gene demonstrated that cyaY is expressed. Finally, comparison of the different polypeptide sequences of ACs permits discussion of important features of the catalytic and regulatory centers of the protein.

OBJECTIVE

To determine the prevalence of celiac disease (CD) in siblings of patients with this disease in Punjab, where wheat is the staple diet.

METHODS

Families of 80 patients with CD diagnosed as per modified ESPGAN criteria were offered family screening. Their siblings aged 2-15 years were tested for serum IgA anti-tissue transglutaminase antibody (anti-tTG) antibody. Those with positive or borderline test and some of those with negative test underwent endoscopic duodenal biopsy. Siblings with characteristics histological findings and showing improvement on follow-up were labeled as having celiac disease.

RESULTS

Of the 63 siblings of 48 index cases studied, 15 tested positive for anti-tTG; of these 13 had celiac disease. Three tested borderline for anti-tTG; none of them had CD. Of the 45 anti-tTG-negative subjects, two agreed to undergo biopsy; one of these had features of CD. Overall, 14 of 63 (22%) siblings had CD, including 8 who had no symptoms suggestive of CD.

CONCLUSIONS

CD is common among siblings of patients with CD in Punjab and may be asymptomatic.

BACKGROUND

Data correlating anti-tissue transglutaminase (tTG) antibody titers with severity of duodenal involvement is limited.

OBJECTIVE

The aim of this study was to correlate IgA anti-tTG antibody titers with symptoms, anthropometric parameters, and duodenal histopathology.

METHODS

Consecutively diagnosed patients of celiac disease as per modified ESPGHAN criteria presenting over a year were enrolled. Demographic data, symptoms, weight-for-age z score (WAZ), height-for-age z score (HAZ), IgA anti-tTG titer, and duodenal histopathology graded as per modified Marsh criteria were recorded. Spearman rank correlation test was used for association between TTG age, WAZ, and HAZ. Receiver operating curve (ROC), sensitivity, specificity, negative predictive value, and positive predictive value were used to obtain anti-tTG cutoff value predictive of Marsh grade 3.

RESULTS

One hundred and forty-two patients with celiac disease were evaluated. tTG showed significant correlation with WAZ (r = 0.822, p = <0.001) and HAZ (r = 0.722, p = <0.001) but not with age (r = 0.202, p = 0.066). The median anti-tTG titers rose progressively with higher Marsh grade on histopathology (p = 0.001). The median anti-tTG titer was also significantly higher in patients with classic celiac disease as compared to non-diarrheal celiac disease (144 u/mL vs. 27, p = 0.02). Anti-tTG titer of 62.5 u/mL was strongly predictive of duodenal histology of Marsh grade 3a and higher with sensitivity, specificity, positive predictive value, and negative predictive value of 95.4 %, 98 %, 93.8 %, and 88.3 % respectively.

CONCLUSIONS

There is a significant correlation between IgA anti-tTG titers and anthropometric parameters and severity of duodenal histopathology. With further validation, strongly positive titers may be sufficient to predict severity of this disease.

BACKGROUND

Several studies have shown the prevalence of celiac disease (CD) to be around 1% in Iran, which is similar to the worldwide prevalence. There is scant information on occult CD in apparently healthy school age children. This study, as the first such study in Iran, aims to determine the prevalence of occult CD in healthy Iranian school age children.

METHODS

In this cross-sectional study, we screened healthy school age children for CD by serum IgA and IgA anti-tissue transglutaminase antibody (tTG) levels. Measurement of these antibodies was by enzyme linked immunosorbent assay. A recheck of positive tTG tests was performed and patients who tested positive underwent endoscopic duodenal biopsies. The biopsy samples were scored according to the Marsh classification by an experienced pathologist.

RESULTS

A total of 634 children (314 males, 320 females; mean age: 12.8 years) were included in the study. All children and/or their parents completed a questionnaire and children underwent an initial physical examination to determine study eligibility. Positive serum tTG was noted in 3 (0.5%; 2 females) out of 634 patients. Duodenal biopsies were consistent with CD in these 3 subjects. The mean age of patients with CD was 14.3 years (range: 12-17 years). The female to male ratio was 2:1. These cases had no signs and symptoms, but a gluten-free diet was recommended according to pathologic changes in their small bowels and results of the tTG test.

CONCLUSIONS

The prevalence of occult CD in these children is 0.5%, which is half of the prevalence of CD in Iranian adults. The anti-tTG concentration at initial serological CD screening is highly informative in determining occult cases of CD. The question is whether all non-symptomatic cases should be treated with a gluten-free diet or not.

BACKGROUND

Celiac disease is a chronic autoimmune enteropathy caused by gluten ingestion. While its prevalence in Western countries is reported to be as high as 1%, the prevalence has not been evaluated in a large-scale study of a Japanese population. The aim of our study was to clarify the possible presence of celiac disease in a Japanese non-clinical population as well as in patients showing symptoms suggestive of the disease.

METHODS

Serum samples were collected from 2008 non-clinical adults and 47 patients with chronic unexplained abdominal symptoms between April 2014 and June 2016. The anti-tissue transglutaminase (TTG) immunoglobulin A antibody titer was determined as a screening test for celiac disease in all subjects, and individuals with a value of >2 U/mL subsequently underwent testing for the presence of serum endomysial IgA antibody (EMA) as confirmation. Those testing positive for EMA or with a high concentration (>10 U/mL) of TTG were further investigated by histopathological examinations of duodenal mucosal biopsy specimens and HLA typing tests.

RESULTS

Of the 2008 non-clinical adults from whom serum samples were collected, 161 tested positive for TTG, and all tested negative for EMA. Four subjects who had a high TTG titer were invited to undergo confirmatory testing, and the histopathological results confirmed the presence of celiac disease in only a single case (0.05%). Of the 47 symptomatic patients, one (2.1%) was found to have a high TTG titer and was diagnosed with celiac disease based on duodenal histopathological findings.

CONCLUSIONS

The presence of celiac disease in a non-clinical Japanese population was low at 0.05% and was rarely found in patients with unexplained chronic abdominal symptoms.

In order to assess the genetic diversity of Mycobacterium tuberculosis in the Novosibirsk Region and determine profiles of resistance, 106 M. tuberculosis isolates were analysed. Fifty (47 %) isolates were identified using variable number tandem repeat typing as being in the Beijing family, of which eight (16 %) were type M2 isolates with the genetic profile 233325153533424 and eight (16 %) were type M11 isolates with the genetic profile 233325173533424, both of which are widespread in Russia. Mutations associated with resistance to isoniazid and rifampicin were identified. Of 48 isolates with resistance to isoniazid, 42 (87.5 %) contained a Ser(315)→Thr substitution in the katG gene and one contained a T→A substitution at position -34 of the promoter region of the ahpC gene. Of 31 isolates with resistance to rifampicin, 19 (61 %) each contained a mutation (TCG→TTG) at codon 531 of the rpoB gene. Two isolates each contained a mutation (GAC→GTC) at codon 516 of the rpoB gene and two others each contained a substitution at codon 526 of the rpoB gene, leading to a His→Asn substitution in one case and a His→Asp substitution in another case. One isolate contained a mutation (CTG→CCG) at codon 533 of the rpoB gene. An association between the Beijing genotype and multidrug resistance was demonstrated (R = 0.2, P = 0.032). However, it was interesting to note that a significant proportion (46 %) of isolates were sensitive to all drugs tested.

To detect the site of mutation in RRDR of rpo B gene for rifampicin resistance in MDR-TB by DNA sequencing. 50 MDR-TB patients were enrolled in our study after informed written consent. Mycobacterial DNA was extracted from sputum samples by Universal Sample Processing (USP) method and RRDR of rpo B gene was amplified by PCR using primers RP4T and RP8T and then sequenced by automated DNA sequencing. The nucleotide sequences of RRDR of rpo B gene were compared with the reference sequence. We observed three different types of mutation in the RRDR of rpo B gene. The frequency of mutation in codon 531 (TCG → TTG), 526 (CAC → TAC) and 516 (GAC → GTC) are 60, 26.6 and 6.6% respectively. Of the total cases studied, 6.6% cases, although resistant to rifampicin, did not show any mutation in the RRDR of rpo B gene. Codon 531 (TCG → TTG) is the most common site of mutation in RRDR of rpo B gene for rifampicin resistance in MDR-pulmonary tuberculosis followed by codon 526 (CAC → TAC) and codon 516 (GAC → GTC).

BACKGROUND

The online version of this article (doi:10.1007/s12291-010-0065-3) contains supplementary material, which is available to authorized users.

Recently we compared the lacI and Hprt mutant frequencies (MFs) and types of mutations in lymphocytes of Big Blue((R)) (BB) rats exposed to 7,12-dimethylbenz[a]anthracene (DMBA) under conditions that result in mammary gland tumors. In this study, we have examined the target mammary tissue for DMBA-induced DNA adducts, lacI MF and types of lacI mutations. Seven-week-old female BB rats were given single doses of 0, 20 or 130 mg/kg DMBA by gavage and the DNA adducts and lacI MFs in the mammary tissue were measured over a period of 14 days and 18 weeks, respectively, following treatment. The lacI MF in the mammary tissue increased for 10 weeks and then remained relatively constant; 130 mg/kg DMBA produced a 14-fold increase in the MF (255 +/- 50 x 10(-6) p.f.u.) over control MF (18. 3 +/- 4 x 10(-6) p.f.u.). (32)P-post-labeling analysis of DNA from mammary tissue and splenic lymphocytes of treated rats revealed two major adducts. Comparison of these adducts with DMBA standards indicated that the adducts formed by DMBA involved both G:C and A:T base pairs. DNA sequencing revealed that the majority of DMBA-induced lacI mutations were base pair substitutions and that A:T->>T:A (44% of the independent mutations) and G:C->>T:A (24% of the independent mutations) transversions were the predominant types. Furthermore, the mutational results revealed a 'hotspot' for a G->>T mutation in codon 95 (GTG->>TTG) of the lacI gene in mammary tissue. These results suggest that DMBA is highly mutagenic to lacI in mammary tissue and that adducts with both G:C and A:T base pairs participate in forming mutations in DMBA-treated BB rats.

BACKGROUND

It has been suggested that high titres of tTG are associated with elevated positive predictive values (PPV) for celiac disease. However, the PPV of a strongly positive tTG will depend on the celiac disease prevalence in the different risk groups of the disease

OBJECTIVE

To assess the PPV of a strongly positive tTG for celiac disease. In addition, to calculate the post-test probability for celiac disease of a strongly positive tTG in a setting of routine clinical practice.

METHODS

145 consecutive celiac disease patients with positive tTG, and with a small bowel biopsy were included. The PPV for different cut-off points of tTG levels for the diagnosis of celiac disease was assessed. In addition, the cut-offs associated with higher PPV were used to calculate the positive likelihood ratio. A simulation in a setting of routine clinical practice was performed to calculate the post-test probability of celiac disease.

RESULTS

No cut-off level was associated with a PPV of 100%. A cut-off of 80 U/mL (11.4×upper normal limit) was associated with the higher PPV value of 98.6%. In the most frequent clinical situations, which in general have a pre-test probability <10%, the post-test probability after having a strongly positive tTG was 90% or less.

CONCLUSIONS

A strongly positive tTG should not be enough to diagnose celiac disease in the most frequent clinical situations, small bowel biopsy remaining as the gold standard in these cases.

OBJECTIVE

Irritable bowel syndrome (IBS) is a common functional gastrointestinal (GI) disorder and coeliac disease (CD) is an auto-immune enteropathy that can mimic almost any functional GI disorder. Both IBS and coeliac disease share common symptoms. The aim of the present study was to estimate the prevalence of CD in patients with IBS and its sub-types.

METHODS

In this cross-sectional study (2008-2010), all consecutive patients with IBS who fulfilled the Rome III criteria attending the GI units in Zahedan (Southeast Iran) were included. Patients based on the sub-type of IBS were classified as diarrhoea-predominant irritable bowel syndrome (D-IBS), constipation-predominant irritable bowel syndrome (C-IBS) and alternating symptoms (mixed type). Immunoglobulin A (IgA) tissue transglutaminase (anti-tTG) was used to screen patients for CD. In the case of positive serologic test, duodenal biopsies were taken to confirm the diagnosis.

RESULTS

A total of 364 (221 females and 143 males) patients with IBS were included. The mean ± standard deviation (SD) age of patients was 37.4 ± 12.4 years. Twenty (5.5%) patients were found to have positive IgA anti-tTG. Main symptoms of patients were diarrhoea (11/20), bloating (10/20) and abdominal distension (6/20). Thirteen (10.5%) patients were found to have positive IgA anti-tTG among the D-IBS, two (1.6%) in the C-IBS and five (4.2%) in M-IBS groups.

CONCLUSIONS

The prevalence of CD in IBS is high. IBS subjects whose main complaint is diarrhoea, bloating or even abdominal distension should be evaluated for CD.

The complete mitochondrial DNA of tube-dwelling diatom, Berkeleya fennica was sequenced and characterized. The circular mitogenome contains 63 genes in 35,509 bp (29.7% GC), including 36 protein-coding, 25 tRNA, 2 rRNA genes. Most of the protein-coding (27) genes have usual ATG start codon, except 9 genes such as ATA for rps8; ATC for rps14; ATT for rps12 and orf51; GTG for nad5; TTA for cox3, nad4 and orf147; and TTG for cob. The nad11 and rrs are the only interrupted genes in the mitogenome. Gene content and synteny of B. fennica are very similar to Phaeodactylum tricoruntum (NC_016739). Absence of repeat region in B. fennica resulted in mitogenome size difference to P. tricoruntum. A new mitogenome will provide useful information for mitochondrial genome diversity and evolution of the diatoms.

All chloroplast genes reported so far possess ATG start codons and sometimes GTGs as an exception. Sequence alignments suggested that the chloroplast infA gene encoding initiation factor 1 in the green alga Chlorella vulgaris has TTG as a putative initiation codon. This gene was shown to be transcribed by RT-PCR analysis. The infA mRNA was translated accurately from the UUG codon in a tobacco chloroplast in vitro translation system. Mutation of the UUG codon to AUG increased translation efficiency approximately 300-fold. These results indicate that the UUG is functional for accurate translation initiation of Chlorella infA mRNA but it is an inefficient initiation codon.

Among the adverse reactions caused by wheat, celiac disease (CD) is the longest studied and best-known pathology. The more recently defined non-celiac gluten sensitivity (NCGS) presents with symptoms which are often indistinguishable from CD. Diagnosis of CD is based on serologic, molecular, and bioptic testing. The IgA anti-transglutaminase (tTG) test is considered highly important, as it shows high sensitivity and specificity and its levels correlate to the degree of intestinal damage. Small bowel biopsy can be avoided in symptomatic patients with IgA anti-tTG levels above 10× the manufacturer's cut-off. Recently, tests of anti-deamidated peptides of gliadin (DGP) have replaced classic anti-native gliadin (AGA) tests. DGP assays have a considerably higher diagnostic accuracy than AGA assays, especially in the IgG class, and can replace anti-tTG tests in patients with selective IgA deficiency. The combination of IgG anti-DGP plus IgA anti-tTG assays show greater sensitivity than a single test, with very high specificity. EMA tests have great diagnostic accuracy but are not recommended by all the latest guidelines because they are observer dependent. Biopsy must still be considered the gold standard for CD diagnosis. HLA-DQ genotyping can be used to screen asymptomatic children and in cases of histology/serology disagreement. About half of NCGS patients are DQ2 positive and have IgG AGA. To diagnose NCGS, first CD and wheat allergy must be excluded; then the wheat dependence of symptoms must be verified by a gluten-free diet and subsequent gluten challenge.

Strong promoters were isolated from Flavobacterium johnsoniae in a promoter-trap vector incorporating a gfp reporter system, and were used to express fluorescent protein markers (including GFP, YFP, mOrange and mStrawberry) and insecticidal protein genes in Flavobacterium strains. Sequence analysis of trapped DNA fragments showed conserved Bacteroidetes promoter motifs (TTG-N(19)-TAnnTTTG) located upstream of putative open reading frames. Plasmids harboring these genomic DNA fragments from F. johnsoniae promoted strong production of fluorescent proteins in Flavobacterium hibernum but not in Escherichiacoli. The most potent promoter (PompA) identified in this work was cloned upstream of genes encoding fluorescent proteins, and these were co-expressed in Flavobacterium strains. The p42 and p51 genes (binary toxins from Bacillus sphaericus) when translationally fused to the 3'-end of gfp showed strong expression. Flavobacteria expressing these genes exhibited toxicity against larvae of the mosquitoes Culex quinquefasciatus, Anopheles gambiae, and Ochlerotatus triseriatus. However, transformants with the transcriptional fusion construct between cry11A with p20 from Bacillus thuringiensis did not express Cry11A protein indicating that constitutive expression of cry11A may be problematic in Flavobacterium.

OBJECTIVE

To establish the prevalence of celiac disease (CD) in girls with Turner syndrome (TS) in British Columbia.

METHODS

Forty-five girls with TS were prospectively screened for CD using blinded testing with the current 'gold standard' - immunoglobulin A (IgA) endomysium antibody (EmA) and the novel IgA tissue transglutaminase antibody (tTG). Those with positive results were offered small bowel biopsies, and a gluten-free diet was recommended if CD was confirmed.

RESULTS

One asymptomatic prepubertal East Indian girl was positive for EmA, had an elevated tTG concentration of 560 U/mL and histological evidence of CD. Seven girls were negative for EmA but had elevated tTG concentrations (175 to 250 U/mL); five were white, one was Asian and one was East Indian. Small bowel biopsies were performed on three girls, and the histologies were normal. The remaining four patients declined biopsy.

CONCLUSIONS

One girl with TS was identified with CD from 45 screened, giving an overall biopsy-confirmed prevalence of 2.2%. This study confirms previous observations placing girls with TS at higher risk for CD and suggests a similar high prevalence in British Columbia.

OBJECTIVE

Transaminasemia develops via different pathways in patients with celiac disease; no information is available on risk factors specifically attributable to celiac disease.

METHODS

We analyzed data collected from consecutive patients referred from January 1997 through December 2009 to the celiac disease clinic at the Spedali Civili of Brescia, Italy. We assessed the factors affecting hypertransaminasemia in 683 patients with celiac disease (based on serologic and biopsy analysis, cohort A; 34 ± 14 years of age) and 304 with functional syndromes (cohort B; 37 ± 13 years of age).

RESULTS

Hypertransaminasemia was detected in 138 patients in cohort A (20%). It was associated with malabsorption (odds ratio [OR], 2.22; P = .004), diarrhea (OR, 1.72; P = .005), and increasing severity of mucosal lesion (Marsh-Oberhuber class; OR, 1.46; P = .001) but not with body mass index (BMI) or the serum level of tissue-transglutaminase antibodies (tTG). Hypertransaminasemia was detected in 22 patients in cohort B (7%) and was associated with the World Health Organization's BMI categories (OR, 7.9; P < .001). In subsets of patients studied with the same analytical method (313 of cohort A and 188 of cohort B), the level of tTG was significantly higher in cohort A at baseline (25.2 ± 16.9 U/L aspartate aminotransferase [AST]) than in cohort B (20.6 ± 9.9 U/L AST, P < .0001) and was related to BMI in cohort B (P = .0012) but not cohort A. When patients were placed on gluten-free diets, the levels of AST decreased from 25.2 ± 16.9 U/L to 19.9 ± 6.6 U/L (P < .0001), independently of the changes of duodenal histology and tTG and correlated with BMI (P = .0007); the prevalence of hypertransaminasemia decreased from 13% to 4%.

CONCLUSIONS

Patients with celiac disease have a higher prevalence of hypertransaminasemia than controls (patients with functional syndromes). Hypertransaminasemia is related to the severity of the duodenal lesion and malabsorption but not BMI. By contrast, there was a positive correlation between the levels of AST and BMI in controls; this relationship was restored when patients with celiac disease were placed on gluten-free diets.