Semen analysis in adolescents with familial Mediterranean fever.

BACKGROUND: Little is known about sperm health in male patients with familial Mediterranean fever (FMF). In this study, the authors aimed to search the frequency of sperm abnormalities of adolescent boys with FMF and also to investigate whether disease activity or colchicine treatment have negative effects on sperm parameters. METHOD: The male adolescents older than 14 years with a diagnosis of FMF were investigated retrospectively. Tel Hashomer and pediatric FMF clinical criteria were used for diagnosis of FMF. Patients who had semen analysis were included in the study. RESULT: Mean age at the diagnosis was 11.13 ± 3.82 years, and mean age at the study was 14.50 ± 0.70 years. The mean sperm concentration was found as 66.26 ± 41.02 million/ml (N > 15 million/ml), the mean total sperm count 113.42 ± 132.39 million (N > 39 million), and the mean sperm motility 51.78 ± 23.70% (N > 40%). Only 8 of 19 (42.1%) patients had normal sperm parameters. Sperm concentration was reduced in two cases, total sperm count was reduced in four patients, and motility was reduced in nine cases. The presence of FMF attacks under treatment was found to be a risk factor for decreased motility in the study group by multivariate regression analysis (odds ratio 0.076, [95% confidence interval 0.005-0.648], P = 0.031). Erythrocyte sedimentation rate at the time of diagnosis was high in patients with low sperm counts compared with those with normal sperm counts (56.00 ± 8.51 vs 24.35 ± 6.32, P: 0.03). Mean colchicine dose at the time of sperm analysis was higher in patients with low sperm motility than that with normal sperm motility (1.72 ± 0.18 vs 1.25 ± 0.08, P: 0.02). CONCLUSION: Sperm abnormalities of male patients with FMF is not infrequent, and it is linked to both inflammation due to uncontrolled disease and colchicine therapy.

[Treated cases of retinopathy of prematurity in Germany : 5-year data from the Retina.net ROP registry]. / Behandelte Frühgeborenenretinopathie in Deutschland : 5-Jahres-Daten des Retina.net ROP-Registers.

BACKGROUND: Retinopathy of prematurity (ROP) is one of the main reasons for childhood blindness. The number of infants requiring treatment, however, is low for individual centers. The Retina.net ROP registry has been founded to allow a joint analysis of treatment patterns and courses post treatment. OBJECTIVE: This paper reports treatment patterns over 5 years. MATERIAL AND METHODS: All infants born between January 2011 and December 2015 who were entered into the treatment registry by one of the 12 participating centers were analyzed. RESULTS: The data of 150 infants (292 eyes) were analyzed and ROP 3+ in zone II was the most prevalent treatment indication. Gestational age and birth weight remained stable over the years. The treatment patterns, however, changed with anti-VEGF treatment (bevacizumab or ranibizumab) accounting for only 10% of treated eyes in 2011 but for 56% and 30% in 2014 and 2015, respectively. Almost all eyes with AP-ROP or zone I disease received anti-VEGF treatment. Zone II disease was predominantly treated with laser photocoagulation. Recurrences were more common and appeared later in the anti-VEGF group compared to the laser group (23%/interval 60 days vs. 17%/interval 23 days). Perioperative complications were evenly distributed across treatment groups. CONCLUSION: The data in this analysis represent about 10-15% of treated infants in Germany. The results provide evidence for an increasing use of anti-VEGF agents for ROP. The data reflect a selection bias for anti-VEGF treatment in eyes with a more aggressive disease. This needs to be considered when interpreting data such as disease recurrence rates. The risk for late recurrences after anti-VEGF treatment is of particular clinical significance.

Renal Transplantation With Size-Matched End-to-End Venous Anastomosis in Children With Inferior Vena Cava Thrombosis.

Due to surgical technical difficulties, inferior vena cava (VCI) thrombosis is contraindicated for renal transplantation in pediatric patients. Of 287 pediatric renal transplantations, 3 patients (9, 12, and 19 kg, respectively) with end-stage renal failure, who had VCI thrombosis at the level of renal vein, underwent end-to-end anastomosis to the proximal aspect of VCI for venous drainage. The latest creatinine values of the patients, who were in the postoperative 56(th), 28(th), and 14(th) months, were 0.6, 0.4, and 0.3 mg/dL, respectively, with graft and patient survival rates of 100%. We think that end-to-end venous drainage into the proximal caval system is the most appropriate surgical approach in pediatric recipients, who have an open suprarenal VCI and a small intra-abdominal cavity, in the presence of an appropriate size-matched graft.

A phase II clinical trial of weekly paclitaxel and carboplatin in combination with panitumumab in metastatic triple negative breast cancer.

PURPOSE: Women with metastatic triple negative breast cancer (TNBC) can have a poor prognosis with treatment limited to cytotoxic chemotherapy. The identification of effective therapies that may limit exposure to cytotoxic chemotherapy and lead to prolonged survival is an unmet medical need. We tested an inhibitor of the epidermal growth factor receptor, panitumumab in combination with chemotherapy. METHODS: We conducted a single arm clinical trial in women with metastatic or locally advanced TNBC to paclitaxel 80 mg/m2 and carboplatin AUC of 2 on days 1, 8, and 15 and panitumumab 6 mg/kg on days 1 and 15 for a cycle length of 28 days. The objectives were to evaluate the response rate and safety of the combination in comparison to historical controls. RESULTS: Fourteen patients with TNBC were enrolled with a median age of 53 years. The majority of women were African American (64.3%) with visceral metastasis (64.2%). Hematologic toxicities, particularly neutropenia and thrombocytopenia, were a major cause of missed chemotherapy and delayed treatment in this study. The overall response rate (complete and partial response) of the 13 evaluable patients was 46%. The median time to best response was 2.4 months and the median time to disease progression was 3.6 months. We were able to perform the PAM50 analysis on tumors from 7 of our subjects. All the samples tested clustered within the basal-like subtype. CONCLUSIONS: In our experience the response rate of carboplatin, paclitaxel and panitumumab was consistent with other reports of response for cytotoxic chemotherapy in metastatic TNBC.

Vitamin D supplementation during the first two years of life in Izmir, Turkey.

AIM: The aim of the present study was to determine the prevalence of vitamin D supplementation and risk factors for non-supplemented infants in Izmir, Turkey. METHODS: This cross-sectional study was carried out in Well-Child Care Clinics in Izmir, Turkey. Parents filled a questionnaire about socio-demographic characteristics and the use of vitamin D. SPSS version 16.0 was used for statistical analysis. RESULTS: A total of 1002 children (aged 1 to 24 months) were enrolled in the study. The supplementation rates of vitamin D were 77% in the first 3 months of life, 57% at 10 to 12 months. Economic status of family, education of parents, occupational status of mothers and parity were associated factors with the use of vitamin D supplements in infants (P<0.05). However, when the data were analyzed using logistic regression analysis, only education of mother seems to be statistically significant independent variable in decreasing non-supplementation/ irregular supplementation. CONCLUSION: Vitamin D supplementation rates seem to be not satisfactory in Izmir. Therefore, the importance of vitamin D supplementation in infants should be emphasized in every well-child care visit to prevent vitamin D deficiency.

The core control system of intracellular iron homeostasis: a mathematical model.

Iron is a metal essential for cellular metabolism. However, excess iron available for reactions contributes to the formation of dangerous reactive oxygen species, such as the hydroxyl radical, via the Fenton reaction. Therefore, intracellular iron levels are tightly constrained by a control system of proteins. This paper contains a mathematical model, in the form of a system of five ordinary differential equations, of the core of this control system, including the labile iron pool as well as proteins that regulate uptake, storage, and export and are connected through negative feedback loops. The model is validated using data from an overexpression experiment with cultured human breast epithelial cells. The parameters in the mathematical model are not known for this particular cell culture system, so the analysis of the model was done for a generic choice of parameters. Through a mixture of analytical arguments and extensive simulations it is shown that for any choice of parameters the model reaches a unique stable steady state, thereby ruling out oscillatory behavior. It is shown furthermore that the model parameters are identifiable through suitable experiments.

A practical method for the determination of sulphur in coal samples by high-resolution continuum source flame atomic absorption spectrometry.

Sulphur in coal was determined using a high-resolution continuum source flame atomic absorption spectrophotometer (HR-CS-FAAS) with actylene/air flame. The C-S absorption band at 258.056 nm was found the most suitable analytical line with respect to sensitivity and spectral interferences. The instrumental parameters were optimized. The coal samples were dried and dissolved using microwave-assisted digestion technique. The validity of the method was tested using standard reference material and certified values were found in the limits of 95% confidence level. Since the concentrations of matrix elements of coal other than carbon are low enough not to cause any spectral interferences, the linear calibration method was applied in all quantifications without any problem. The calibration standards were prepared in sulphuric acid. The method was accurate, fast, simple and sensitive. The limit of detection (LOD, 3δ, N=10) and the limit of quantification (LOQ, 10δ, N=10) were found to be 0.01 and 0.03% (w/w), respectively. The sulphur concentrations of various kinds of the coal samples received around Turkey were determined. The sulphur contents of the coal samples were ranged from ≤ LOQ to 1.2%.

Diverse genotypical features and impacts on clinical course and severity of cystic fibrosis: early childhood experience.

AIM: Very little is known about the relationship between genotype and phenotype of cystic fibrosis (CF) from the Turkish children. The aim of the study was to analyze the genotype and phenotype of 24 children with CF and to investigate the correlation between type of mutation in cystic fibrosis transmembrane conductance regulator (CFTR) protein gene and clinical manifestation of the disease. METHODS: Patients were evaluated retrospectively and prospectively. History, clinical findings, sweat test and mutation analysis were used for the definitive diagnosis of CF. Phenotypical features of 24 cases were evaluated according to clinical findings. We compared the clinical phenotype and age at diagnosis, genotypic features. A total of 36 mutations were analyzed by polymerase chain reaction (PCR) and reverse hybridization methods. Statistical analysis was done by using χ2, Fisher exact and Pearson correlation tests. RESULTS: The mean age of the cases that were admitted to our out-patient clinic was 5.3±4 years. The median age of diagnosis was three months. Parents were consanguineous in 37.5% of cases and loss of a sibling at one year of age was stated in a quarter. The most frequent symptom was recurrent diarrhoea (79.2%) and there was severe growth retardation in 12 (50%) and pseudo-Bartter (PB) syndrome in 11 of the cases. The incidence of PB was higher in cases that were diagnosed at one year of age. Out of 18 cases with mutation analysis, nine (50%) were positive for DF508 mutation, and four cases were homozygous out of nine cases. Two separate mutations were determined in two cases with severe clinical picture. The incidence of respiratory tract infection during the admission was lower in DF508 positive cases (P=0.016). There was no statistically significant relation between DF508 positivity and diarrhea, severe growth retardation and PB (P>0.05). The other mutations that were determined in our patients were rarely seen mutations such as 3120+1 G-A, R347P, 1677delTA, 2789+5G-A, 2183AA-G, and R1066C. CONCLUSION: DF508 mutation rates in our cases diagnosed in early childhood were higher than the rates reported previously in Turkish children. The definition of molecular defect in CFTR gene has an impact on verifying the diagnosis and decreasing morbidity and mortality. An adequately large sample size is needed to evaluate the mutation profiles and genotype-phenotype characteristics in our country.

A case report: hepatic posttransplant lymphoproliferative disorder in a non-liver transplant patient.

Posttransplant lymphoproliferative disorder (PTLD) is the most common malignancy in children after solid organ transplantation. We present a patient, who developed Epstein-Barr virus (EBV)-related PTLD in the liver after renal transplantation. A 10-year-old EBV-seronegative boy with cystinosis underwent a living related preemptive renal transplantation. He received antiviral prophylaxis with valacyclovir. At 5.5 months posttransplantation he displayed a primary EBV infection with an high fever, hepatosplenomegaly, monocytosis, and positive EBV DNA levels. Two months there after, a hypoechoic nodular 20-mm lesion in the left lobe of liver was detected on abdominal ultrasonography, performed because of anorexia and weight loss. EBV-DNA copy number was 7820 copies per milliliter. Liver biopsy showed a diffuse large B-cell lymphoma that was compatible with PTLD. We stopped all immunosupressive agents other than prednisolone. Chemotherapy consisting of two courses of cyclophosphamide, vincristine, prednisolone, and adriamycin was followed by rituximab. Within 2 months, the lesion resolved and within 18 months, he was free of disease.

Influence of several fibre-reinforced composite restoration techniques on cusp movement and fracture strength of molar teeth.

AIM: To compare mean cusp movement in molar teeth with endodontic access and mesial-occlusal-distal (MOD) cavities before and after restoration with several fibre-reinforced composite restoration techniques under loading and to evaluate the effect of restoration technique on fracture strength. METHODOLOGY: Reference points were marked at the mesial cusp ridges of extracted human mandibular molar teeth. Digital images were taken under loading (300 N) using a stereomicroscope (Leica MZ16A; Wetzlar, Germany). Three-dimensional (3D) distances between the reference points were recorded (Leica, Stereo-Explorer, 2.1) as controls. Standard MOD cavities were prepared and restored as follows (n = 10), group 1: composite restoration (Clearfil AP-X; Kuraray, Tokyo, Japan); group 2: cavity lined with polyethylene fibre (Ribbond, Ribbond Inc., Seattle, WA, USA) in combination with flowable resin (Protect-Liner F; Kuraray, Tokyo, Japan) before composite restoration; group 3: polyethylene fibre inserted on occlusal surface of the tooth from buccal to lingual after finishing the composite restoration; group 4: missing walls were restored with composite resin and inner surfaces of the axial walls were then reinforced with polyethylene fibre placed circumferentially before the composite restoration. The restored teeth were re-loaded, digital images were re-taken and the 3D distance between the reference points was recorded in µm. Comparisons of the restoration techniques, the effectiveness of restoration for each group were analysed statistically (Kruskall-Wallis, paired-samples t-test). The teeth were then loaded until failure (5 mm min(-1) ), the data were recorded (N) and analysed statistically (Kruskall-Wallis test). RESULTS: A significant difference occurred amongst the groups in terms of cusp movement (P = 0.018). All the groups revealed a decrease in inter-cuspal width when compared to their initial records. The mean values of these decreases were as follows: group 1 17.6 (P = 0.003), group 2 6.7 (not sig), group 3 6.6 (not sig) and group 4 0.85 (not sig) µm. No significant difference was found amongst the fracture strength values (P = 0.22). In group 1, 90% of the fractures were non-restorable, whereas in group 3 100% of the fractures were restorable. CONCLUSIONS: Regardless of restoration technique, fibre reinforcement of composite restorations decreased cusp movement in molar teeth with MOD and endodontic access cavities but did not affect fracture strength.

Slurry analysis of cadmium and copper collected on 11-mercaptoundecanoic acid modified TiO2 core-Au shell nanoparticles by flame atomic absorption spectrometry.

Separation/preconcentration of copper and cadmium using TiO(2) core-Au shell nanoparticles modified with 11-mercaptoundecanoic acid and their slurry analysis by flame atomic absorption spectrometry were described. For this purpose, at first, titanium dioxide nanoparticles were coated with gold shell by reducing the chloroauric acid with sodium borohydride and then modified with 11-mercaptoundecanoic acid. The characterization of modified nanoparticles was performed using ultra-violet spectroscopy and dynamic light scattering. Copper and cadmium were then collected on the prepared sorbent by batch method. The solid phase loaded with the analytes was separated by centrifugation and the supernatant was removed. Finally, the precipitate was slurried and directly aspirated into the flame for the determination of analytes. Thus, elution step and its all drawbacks were eliminated. The effects of pH, amount of sorbent, slurry volume, sample volume and diverse ions on the recovery were investigated. After optimization of experimental parameters, the analytes in different certified reference materials and spiked water samples were quantitatively recovered with 5% RSD. The analytes were enriched up to 20-fold. Limits of detection (N=10, 3σ) for copper and cadmium were 0.28 and 0.15 ng mL(-1), respectively.

The effect of different posts on fracture strength of roots with vertical fracture and re-attached fragments.

The aim of this in vitro study was to test the effect of different post systems on fracture strength of roots with re-attached fragments. Root canals of eighty extracted single-rooted human teeth were instrumented (ProFile) and randomly divided into two groups. The roots in the first group were vertically cracked, and the fragments were re-attached using Super Bond C&B (Sun Medical, Tokya, Japan). The roots in the second group were kept sound. Obturation of the roots was performed with MetaSEAL (Sun Medical) and gutta-percha. Post spaces were prepared, and the roots were restored with one of the followings: UniCore (Ultradent), Everstick (Stick Tech), Ribbond (Ribbond), ParaPost (Coltene/Whaledent) (n = 10). Four mm high build-ups were created (Clearfil DC Bond Core; Kuraray, Tokyo, Japan). Compressive loading of the samples was performed after 24 h (1 mm min(-1)). Mean load necessary to fracture each sample was recorded (Newton) and statistically analysed (One-way anova, t-tests). ParaPost showed the highest fracture strength among the roots with re-attached fragments (P < 0.05). UniCore and ParaPost systems showed similar fracture strength in the sound roots (P > 0.05). Re-attached fragments significantly reduced the fracture strength of roots in UniCore group (P = 0.000). Ribbond post showed mostly repairable fractures. Metal post (ParaPost) showed the highest fracture strength in the roots with re-attached fragments; however, fracture pattern was 41% non-repairable. Re-attached fragments significantly reduced the fracture strength of the roots in UniCore group. Prefabricated posts showed similar fracture strength in the sound roots. Customized post systems EverStick and Ribbond showed mostly repairable failure after loading in sound roots or roots with re-attached fragments.

A novel slurry sampling analysis of lead in different water samples by electrothermal atomic absorption spectrometry after coprecipitated with cobalt/pyrrolidine dithiocarbamate complex.

A preconcentration/separation technique based on the coprecipitation of lead with cobalt/pyrrolidine dithiocarbamate complex (Co(PDC)(2)) and subsequently its direct slurry sampling determination by electrothermal atomic absorption spectrometry (AAS) was described. For this purpose, at first, lead was coprecipitated with cobalt/pyrrolidine dithiocarbamate complex formed using ammonium pyrrolidine dithiocarbamate (APDC) as a chelating agent and cobalt as a carrier element. The supernatant was then separated and the slurry of the precipitate prepared in Triton X-100 was directly analyzed by electrothermal atomic absorption spectrometry with respect to lead concentration. The effects of experimental conditions on coprecipitation of lead with gathering precipitate as well as homogeneity and stability of the slurry were investigated. After the optimization of experimental parameters, a 100-fold enrichment of the analyte with quantitative recovery (>90%) and high precision (<10% R.S.D.) were obtained. By using the proposed technique, the lead concentrations in heavy matrices of Certified Sea-water and wastewater samples could be practically and rapidly determined in the range of 95% confidence level. The detection limit of the described method for lead using sample-matching blanks was 1.5 ng/L (3 sigma, N=10).

Survey of lead and copper in Turkish raisins.

Lead and copper levels in various types of Turkish raisins, collected from the most important production centers, were determined by electrothermal atomic absorption spectrometry. Samples were principally the products of 2005; however, two different and important raisin types produced between 2003 and 2006 were also analyzed. To investigate the source of pollution, the lead and copper content of all samples were separately determined after successive treatment with water, then with acetone and, finally, complete decomposition in a HNO(3)/H(2)SO(4)/H(2)O(2) mixture. Metal levels in raisin seeds were also determined. The results were statistically evaluated, indicating that the raisins taken from different regions and years contained a mean (range) of 0.056 (0.012-0.359) mg kg(-1) lead and 2.542 (0.770-4.706) mg kg(-1) copper. Lead concentrations in Turkish raisins were significantly lower than those found in a previous study.

Outcome of pediatric liver transplant recipients in Turkey: single center experience.

To summarize the evolution of the pediatric liver transplant program in a developing country. Between April 1997, and September 2003, 32 cadaveric (CD) and 35 living donor (LD) liver transplantations were performed on 61 children (median age 3.8 yr, range 0.5-16) at Ege University Organ Transplantation and Research Center. The patient's charts were reviewed retrospectively. The outcome of patient and graft survival was analyzed and the incidence of graft loss, complications and rejections was calculated. Indications for liver transplantation were metabolic liver disease (n = 17), biliary atresia (n = 14), viral hepatitis (n = 4), autoimmune hepatitis (n = 6), cryptogenic cirrhosis (n = 11), fulminant liver failure (n = 5) and others (n = 5). Seven of 61 children with chronic liver disease had hepatocellular carcinoma concomitantly. Median pediatric end-stage liver disease score was 23 (range 1-54). Seven children (11.4%) were UNOS status I, 44 (72%) were UNOS status II and 10 (16.6%) were UNOS status III. The median follow-up of the study population was 3.6 yr (range 0.5-6). Actuarial patient survival rates at 1, 2, 3 and 4 yr were 86, 86, 71.3 and 65% in the CD group vs. 80, 76, 67 and 67% in the LR group, respectively (p = NS). Patients listed as UNOS status 1 had lower survival rates than patients listed as UNOS status 2 and 3 (p < 0.05). The mortality rate was 26.2%. Graft survival rates were 81, 81, 75 and 64% at 1, 2, 3 and 4-yr respectively. Six patients (9%) underwent retransplantation. The main complications were infections (64.7%) and surgical complications (43.2%) (including biliary complication, vascular problems, postoperative bleeding, small for size and large for size). The incidence of acute cellular rejection was 39.3%, whereas chronic rejection was 7.4%. The result of liver transplantation in Turkish children was slightly inferior to those reported for North American and European children. However, an important characteristic of these patients that distinguishes them from Europe and North America is that most were UNOS status IIa and UNOS status I (44%). Despite technical and medical progress, infectious and biliary problems have continued to be an important cause of mortality in these patients.

The interference effect of a mixture of magnesium, aluminium, sulfate and chloride on the atomization and vaporization of manganese in graphite furnace atomic absorption spectrometry.

In this study, the interference effects of Al3+, Mg2+, Cl- and SO4(2-) ions on the determination of manganese by graphite furnace atomic absorption spectrometry (GFAAS) were investigated. At first, the interferences caused by Al2(SO4)3, AlCl3, MgCl2 and MgSO4, which are the most possible major compounds for the combinations of the ions mixed, were individually considered. Then, the effects caused by mixtures containing various amounts of MgSO4 and AlCl3 were studied. If the pyrolysis temperature is below 800 degrees C, AlCl3 changes the vaporization mechanism of manganese. These interferences disappear at higher pyrolysis temperatures. At the same time, aluminum salts may cause the formation of refractory compounds between aluminum and manganese (like spinel MnAl2O4) that shift the absorption signals of manganese to higher temperatures. Magnesium sulfate, by itself, does not cause any depression of manganese signals. In fact, it acts as a modifier, preventing volatilization losses of manganese during the pyrolysis step. A conclusion was reached that detailed investigation of the interferences in a complex media is a very difficult experimental and theoretical task. To solve practical problems, one may better follow the general notions developed in GFAAS toward complex matrices.

An outbreak of Aseptic Meningitis due to echovirus type 30 in two cities of Turkey.

An outbreak of aseptic meningitis due to echovirus 30 occurred in Ankara and Antalya in Turkey, during June to September 1999, with 176 cases fitting the clinical case definition. Cases were ascertained from attendees of the three hospitals in Ankara and one in Antalya. Medical files were reviewed and evaluated retrospectively. Clinical presentation and laboratory findings were typical of viral meningitis. Cerebrospinal fluid and/or stool samples of 86 patients were cultured for enteroviruses. Echovirus type 30 was isolated in 38 patients (44%). This is the first report and epidemiologic data about an aseptic meningitis outbreak due to echovirus type 30 from Turkey.

Association between anti-endomysial antibody and total intestinal villous atrophy in children with coeliac disease.

BACKGROUND: There is growing evidence to suggest that detection of anti-gliadin antibody (AGA) and anti-endomysial antibody (EmA) can serve as sensitive markers of the degree of histological abnormalities in patients with coeliac disease. AIM: To evaluate the association between the presence of AGA and EmA and villous atrophy in intestinal biopsies of children with suspected coeliac disease. SETTINGS AND DESIGN: Intestinal samples of 46 children with failure to thrive, chronic diarrhoea, malabsorption and short stature with either AGA and/or EmA positivity were evaluated, retrospectively. The diagnosis of coeliac disease was based on ESPGHAN criteria. METHODS AND MATERIAL: Patients with total villous atrophy who fulfilled the ESPGHAN criteria for the diagnosis of coeliac disease were diagnosed to have coeliac disease. Nine patients without villous atrophy were taken as negative controls for this study. AGA-IgA was measured both by immunoflourescence (IF) and ELISA and EmA-IgA by IF while patients were on normal diet. Relationship between autoantibody positivity and intestinal total villous atrophy was evaluated. RESULTS: Overall positivity for AGA IgA was 85% (39/46) by IF+ELISA and EmA positivity was 85% (39/46) by IF within the study group. Histological examination revealed total villous atrophy with lymphocyte infiltration and crypt hyperplasia in 37 (80%) patients. AGA IgA was positive in 14 (38%) and 31 (84%) of these children by ELISA and IF, respectively. EmA positivity was detected in 35/37 (95%) cases with atrophy and 4/9 (44%) without atrophy (p=0.002). Thirty out of 37 (81%) patients with villous atrophy had both AGA IgA (IF) and EmA positivity (p=0.186). All of the sixteen patients that had both positive AGA IgA (ELISA+IF) and EmA had total villous atrophy (p=0.037). CONCLUSION: A significant association between total villous atrophy and EmA positivity has been documented in this study.

A new assay to quantify in vivo repair of G:T mispairs by base excision repair.

The double mismatch reversion (DMR) assay quantifies the repair of G:T mispairs exclusively by base excision repair in vivo. Synthetic oligonucleotides containing two G:T mispairs on opposite strands were placed into the suppressor tRNA gene supF in the shuttle plasmid pDMR. Placement of two mispairs on opposite strands of supF creates a one to one correspondence between the number of correct repair events prior to replication in which G:T mispairs are converted to G:C base pairs and the number of post-replication progeny plasmids with functional supF. Replication of unrepaired or incorrectly repaired mispairs cannot produce progeny plasmids containing functional supF. Indeed, direct transformation of Escherichia coli strain MBL50, which reports the functional status of supF, with pDMR constructs containing two G:T or G:G mispairs yielded <0.5% wild-type supF-containing colonies. In contrast, passage of G:T mispair-containing pDMR constructs through human 5637 bladder carcinoma cells for 48h prior to plasmid recovery and transformation of the reporter E. coli strain MBL50 produced 47% wild-type supF-containing colonies. This finding was indicative of repair prior to the onset of replication in 5637 cells. However, passage of G:G mispair-containing pDMR constructs through 5637 cells yielded <0.5% wild-type supF-containing colonies. Moreover, no difference was observed in the rate of G:T mispair repair by HCT 116 colorectal carcinoma cells deficient in long-patch mismatch repair and a long-patch mismatch repair proficient HCT 116 subline. These data demonstrate that repair measured by the DMR assay is exclusively attributable to short-patch pathways. The DMR assay proved useful in the analysis of the effect of the base 5' to a mispaired G on the rate of G:T base excision repair by 5637 cells, indicating the sequence preference CpG approximately 5mCpG>TpG>GpG approximately ApG, and in the comparison of G:T base excision repair rates between cell lines.