Does Infliximab Influence Surgical Morbidity of Ileal Pouch-Anal Anastomosis in Patients with Ulcerative Colitis?

Purpose Since infliximab has been approved for treatment of patients with refractory ulcerative colitis, surgeons will be increasingly faced with operating on patients who have failed therapy with this potent immunosuppressant. This study was designed to compare short-term complications in patients with ulcerative colitis who were treated with and without infliximab before colectomy. Methods The charts of patients undergoing ileal pouch-anal anastomosis or subtotal colectomy for refractory ulcerative colitis during the five-year period ending October 2005 were reviewed. Postoperative medical and surgical complications were assessed. Results Seventeen patients had failed infliximab treatment and 134 patients were never treated with infliximab. Ileal pouch-anal anastomosis was performed in 112 patients (74 percent) and subtotal colectomy in 39 patients (36 percent). There were no deaths. Postoperative complications were observed in 43 patients (28 percent), with no significant difference observed between infliximab-treated (37 percent) and infliximab-untreated patients (27 percent). Of 61 patients (40 percent) treated with preoperative cyclosporine A, 5 patients also had been treated with infliximab. The infliximab and cyclosporine A-treated patient group had an 80 percent complication rate, significantly higher than the 29 percent complication rate noted in the cyclosporine A only-treated group (P = 0.04). Conclusions Although preoperative treatment with infliximab alone does not significantly increase the incidence of postoperative complications, using both inflixiamb and cyclosporine A before colectomy in refractory ulcerative colitis is associated with high surgical morbidity. Read at the meeting of The American Society of Colon and Rectal Surgeons, Seattle, Washington, June 3 to 7, 2006. Reprints are not available.     

Bile Acid Malabsorption as a Cause of Chronic Diarrhea Diagnostic Value of 7α-Hydroxy-4-Cholesten-3-One in Serum

To evaluate the usefulness of7-hydroxy-4-cholesten-3-one (HCO) serumconcentrations as a diagnostic marker of bile acidmalabsorption, we determined the reference range of HCOin 106 normal subjects (age 40.2 ± 16.8 years; 55 women, 51men) and conducted a utility study in 23 patients withchronic diarrhea of unknown origin (age 49.4 ±15.3 years, 13 women, 10 men). The diagnosis of bileacid malabsorption was made on the basis of a decreased retentionof [⁷⁵Se]homocholyltaurine after oralapplication (⁷⁵SeHCAT test). HCO (referencerange: 6-48 ng/ml) and the ⁷⁵SeHCAT testyielded the same results in 19/23 (83%) patients. Bile acid malabsorption wasidentified by an increase of HCO in serum with asensitivity of 90% and a specificity of 79%. Analysis ofHCO in serum may serve as a novel, simple, and sensitive method for the detection of bile acidmalabsorption in patients with chronic diarrhea ofunknown origin.     

Determinants of Osteopenia in Male Renal-Stone–Disease Patients with Idiopathic Hypercalciuria

Summary

Background and objectives

Bone demineralization is frequent in renal-stone formers with hypercalciuria. Although this pathologic link has been recognized for decades, the underlying mechanisms and risk factors associated with osteopenia/osteoporosis in this population remain partially understood.

Design, setting, participants, & measurements

This study retrospectively analyzed determinants of low bone mineral density (BMD) in 65 idiopathic hypercalciuric male renal-stone formers. Clinical and biologic evaluation included BMD measurement, bone-remodeling markers, analysis of calcium metabolism with oral calcium load test, and dietary inquiry.

Results

Patients with osteopenia (n = 23, 35% of the population) presented significantly higher fasting calciuria as compared with normal bone density patients (n = 42) (calcium/creatinine ratio was 0.32 versus 0.24 mmol/mmol; P = 0.006). Analysis of the whole population revealed a negative association between fasting hypercalciuria and BMD (P = 0.003), independent of confounding variables including body-mass index and tobacco consumption. The fasting calcium/creatinine ratio above 0.25 mmol/mmol was associated with a 3.8-fold increase in the risk of low BMD.

Conclusion

In our study, fasting hypercalciuria after a 2-day calcium-restricted diet appears as the only biologic factor associated with low BMD, suggesting a bone-calcium efflux. Our results support the view of a parathyroid-independent pathologic process that remains to be identified. Hypercalciuric patients with low BMD do not excrete more calcium in 24-hour urine samples than patients without low BMD.     

Frequency of α-Globin Gene Triplications and Coinheritance with β-Globin Gene Mutations in the Iranian Population

Abstract

Numerical variation in α-globin genes is very important due to their roles as an effective factor for phenotype presentation. An unequal crossover from misalignment of a homologous sequence of an α-globin gene during meiosis can produce a numerical alteration. A single α-globin gene deletion is the most frequent mutation in α-thalassemia (α-thal) worldwide, while the additional α-globin chain is relatively common. The excess α-globin gene plays a critical role in pathophysiology of thalassemia, especially when in coinherited with β-thalassemia (β-thal). α-Globin triplication leads to an imbalanced ratio between α- and β-globin chains, thus, it can exacerbate the clinical and hematological features of β-thal. Different studies have been performed in various countries to determine the frequency of α-globin triplication and its genotype-phenotype correlation with β-thal. In this study, we focused on the frequency of α-globin gene triplication and its characterization, either solely or in coexistence with β-globin gene mutations in Iranian populations. We have investigated the α-globin gene rearrangements in 4010 individuals from different provinces of Iran with normal to abnormal hematological parameters. In total, the frequency of the ααα^{anti 3.7} triplication was 1.7% and phenotype aggravation was observed in α-globin triplication patients who were carriers of β-thal. Therefore, identification of genotype-phenotype correlation of α-globin triplication with β-thal can be very useful for predicting the severity of clinical manifestations during genetic counseling.     

Association of IFN-¹ Gene Polymorphism with Type 1 Diabetes in Iranian Patients

Background: Type 1 Diabetes (T1D) is a chronic and progressive autoimmune disorder. Cytokines play a critical role in the pathogenesis of T1D.   Objective: IFN-¹ polymorphism was investigated in T1D and compared with normal controls.   Methods: Thirty patients suffering from T1D and 40 normal controls were studied simultaneously using PCR technique. IFN-   ¹ gene was evaluated at position 5’UTR +5644. Results: There was a significant difference between patient and control groups in TT genotype (P<0.05). Conclusion: In this study, we found a negative association between IFN-¹ gene at position 5’UTR +5644 and T1D in Iranian patients pointing to T allele as a protective factor against T1D.     

Increased Levels of γGT Suggest the Presence of Bile Duct Lesions in Patients with Chronic Hepatitis C

Damage to bile ducts in chronic hepatitis C is a characteristic histological lesion. Moreover, the presence of abnormal levels of gammaGT in these patients is also a common finding. Assessing whether the presence of bile duct lesions is indicated by biochemical abnormalities or whether virological characteristics can influence their development may help in the definition of clinical-histological relationships in chronic hepatitis C. In this study we evaluated the relationships among routine biochemical parameters, serum bile acids, and pi-class glutathione S-transferase levels, and the presence of bile duct lesions in 60 patients with chronic hepatitis C. Furthermore, we assessed whether the presence of bile duct lesion might be related to HCV genotype, HCV-RNA serum levels, and positivity for HGV-RNA. We found that gammaGT was the only parameter related to the presence of bile duct lesions. Although a trend towards higher serum bile acids and pi-class glutathione S-transferase levels was observed in patients with bile duct lesions, this trend did not reach statistical significance. Different HCV genotypes and RNA levels, and HGV-RNA positivity did not seem to influence the presence of bile duct damage. In conclusion we found that gammaGT levels point out the presence of bile duct lesions in patients with chronic hepatitis C. Since we observed a different pattern of alteration of gammaGT, serum bile acids, and pi-class glutathione S-transferase, we suggest that these various biochemical alterations reflect a more complex damage to bile duct structures, which is not likely represented by the common assessment of bile duct lesions. Viral factors such as HCV genotype and RNA levels as well as HGV-RNA positivity are probably not the main cause of this histological damage.     

Increased Frequency of Mutations in the Gene Responsible for Familial Mediterranean Fever (MEFV) in a Cohort of Patients with Ulcerative Colitis: Evidence for a Potential Disease-Modifying Effect?

The MEFV gene, responsible for familial Mediterranean fever (FMF), is involved in inflammatory reactions through altered leukocyte apoptosis, secretion of interleukin (IL)-1β, and activation of the NF-κ B pathway. Ulcerative Colitis (UC) and FMF are both characterized by a recurrent pattern of presentation with periods of remission and flares associated with neutrophilic infiltration at the site of injury. The aim of this study was to investigate the possible correlation between UC and MEFV gene alterations. Twenty-five consecutive, first-diagnosed and untreated UC patients, 28 control patients with rheumatoid arthritis, and 65 normal individuals were analyzed. Nonisotopic RNase Cleavage Assay (NIRCA) was applied as a first-step mutational screening method of exons 10 and 2 of MEFV gene; direct sequencing was subsequently performed to confirm the results. MEFVmutations were identified in 7 (3 M694V/0, 2 M680I/0, 1 E148Q/E148Q, and 1 A744S/0) out of 25 UC patients versus 1 (M694V/0) out of 28 rheumatoid arthritis patients (P = .0199) and 1 (M694V/0) out of 65 healthy controls (P = .0004). Four out of 7 patients with MEFVmutations had inflammatory arthritis, a clinical finding that was not observed in the 18 UC patients with unmutated MEFV (P = .0028). Patients with UC almost universally carried the T A C G MEFV exon 2 haplotype in contrast with normal individuals (P < .0001) and FMF patients (P = .0310). In conclusion the increased frequency of mutations of MEFV in UC patients, especially in those with episodic arthritis, suggests a possible modifying effect of MEFV in the disease process and its localization within the joint. The difference in distribution of MEFV exon 2 haplotypes between UC patients and both FMF patients and normal individuals, suggests that UC patients constitute a genetically distinct population. Larger, longitudinal studies are needed to confirm these initial findings.     

Clinical Features and Quality of Life in Patients with Different Phenotypes of Crohn’s Disease of the Ileal Pouch

Purpose Crohn’s disease of the pouch can occur in patients with colectomy and ileal pouch-anal anastomosis performed for ulcerative colitis. The clinical features of inflammatory, fibrostenotic, and fistulizing Crohn’s disease have not been characterized. Methods A total of 73 eligible patients with Crohn’s disease of the pouch, who were seen in the Pouchitis Clinic, were enrolled: 25 with inflammatory Crohn’s disease, 17 with fibrostenotic Crohn’s disease, and 31 with fistulizing Crohn’s disease. The clinical phenotypes of Crohn’s disease were based on a combined assessment of clinical, endoscopic, radiographic, and histologic features. Clinical symptoms, endoscopic and histologic features, and health-related quality-of-life scores were assessed. Results Demographic and clinical features, including preoperative and postoperative parameters, were similar between the three phenotypes of Crohn’s disease of the pouch. The use of nonsteroidal anti-inflammatory drugs, neuropsychiatric drugs, antidiarrheal agents, and Crohn’s disease medicines was not different between the three groups. Predominant symptoms, as expected, were significantly different between the three phenotypes: diarrhea and/or pain in 92 percent of patients with inflammatory Crohn’s disease, obstructive symptoms in 64.7 percent of patients with fibrostenotic Crohn’s disease, and fistular drainage in 51.6 percent of those with fistulizing Crohn’s disease (P < 0.0001). There was no statistical difference in quality-of-life scores between the three phenotypes, adjusted for disease activity. There was no significant correlation between quality-of-life and symptom scores in any of the three groups. Although not statistically significant, patients with fistulizing Crohn’s disease (16.1 percent) tended to have an increased risk for pouch failure compared with inflammatory (8 percent) or fibrostenotic (5.9 percent) Crohn’s disease. Conclusions Predominant symptoms were different in clinical phenotypes of Crohn’s disease. Each of the three phenotypes of Crohn’s disease similarly affected quality-of-life. Fistulizing Crohn’s disease may be associated with a higher risk for pouch failure. Supported by NIH R03 DK 067275 and an American College of Gastroenterology Clinical Research Award. Reprints are not available.     

p53 Mutations are Associated with Dysplasia and Progression of Dysplasia in Patients with Crohn’s Disease

Background: Mutations in the tumor suppressor gene p53 are associated with neoplasia in ulcerative colitis, but little is understood of their significance in Crohn’s disease (CD). Purpose: To explore p53 expression as a marker of neoplasia in CD patients. Methods: This is a retrospective review of CD patients who underwent p53 IHC staining in our center between 1995 and 2003. The p53 status was correlated to the presence and grade of neoplasia at the time of staining and in subsequent follow-up. Results: Fourteen CD patients had p53 assessment: eight were p53 positive and six were p53 negative. Seven of eight p53+ had dysplasia (six LGD, one HGD); one of six p53−had dysplasia (LGD) (P = 0.03). Four p53+ patients with follow-up had persistent dysplasia and two had progression to a higher grade. Three p53− patients with follow-up remained free of dysplasia. Conclusions: This limited study shows that p53 over expression in CD patients is associated with dysplasia that may progress to a higher grade of neoplasia over time.     

Krüppel-Like Factor 1 Gene Mutations in Thalassemia Patients from North Iran: Report of a New Mutation Associated with β-Thalassemia Intermedia

Thalassemia is a hereditary disease with an autosomal recessive inheritance pattern resulting in reduced production of globin chains. Mutations in modifier genes can cause or affect thalassemia. Krüppel-like factor 1 (KLF1) is a modifier gene that was investigated in this study. Thirty-five Iranian β-thalassemia (β-thal) minor patients with hematological symptoms including Hb A₂ 3.0%, mean corpuscular volume (MCV) <75.0 fL, mean corpuscular hemoglobin (Hb) (MCH) <25.0?pg, and two β-thal intermedia (β-TI) patients in 50 subjects who carried no mutations on the HBB and HBA2 or HBA1 genes were investigated for all exons of the KLF1 gene by polymerase chain reaction (PCR) and sequencing methods. Of the 35 patients with a β-thal minor phenotype, one patient was heterozygous for the c.544T>C mutation in exon 2 of KLF1 and HBB: c.380T>G variant, Hb Dhonburi [also known as Hb Neapolis or codon 126 (T>G)]. The c.340T>C mutation was also found in exon 2 of the KLF1 gene with an allele frequency of 16.6% in the studied β-thal carriers. The two β-TI patients were homozygous for a new mutation c.942delA in exon 3 of KLF1. Mutations in modifier genes can cause or affect thalassemia. Therefore, exact investigation of globin genes and modifiers such as KLF1 is necessary in areas where globin gene disorders are most prevalent to understand the reason of clinical and hematological symptoms of thalassemia and facilitate newborn screening or prenatal diagnosis (PND) programs.     

Novel and Recurrent Mutations of STK11 Gene in Six Chinese Cases with Peutz–Jeghers Syndrome

Background

The serine/threonine kinase 11 (STK11) gene is the main causal gene in Peutz–Jeghers syndrome (PJS). Abnormal STK11 may increase cancer risk of PJS patients via affecting its target proteins such as P53, AMPK, and PTEN. In this study, we investigated the molecular basis of six Chinese PJS patients.

Materials and Methods

Blood samples were collected from four Chinese PJS families and two sporadic patients. The entire coding region of the STK11 gene was amplified by polymerase chain reaction and analyzed by direct sequencing. Functions of mutants were assessed by PolyPhen-2, Swiss-Model software, and luciferase reporter assay.

Results

Novel mutations (c.842_843insC, c.804_805insG, and c.922T>G) and recurrent mutations (c.526G>A, c.180C>G, and c.1062C>G) were identified. Missense mutation c.922T>G and c.526G>A were predicted as probably damaging by PolyPhen-2, while c.1062C>G was benign. Mutation c.108C>G was a nonsense mutation. The 284Ter mutants of c.842_843insC and c.804_805insG significantly diminished the capacity of P53 activity in 293FT cells.

Conclusions

Our results support that STK11 gene mutations underlie Chinese patients with PJS. Mutation involving partial kinase domain disrupts normal function of STK11. Our results also enlarge the spectrum of STK11 variants in PJS patients.     

Gene Expression of Atrial Calcium-Handling Proteins in Patients with Rheumatic Heart Disease and Atrial Fibrillation

Objectives To investigate the gene expression of calcium - handling proteins in patients with rheumatic heart disease (RHD) and atrial fibrillation (AF) . Methods A total of 50 patients with rheumatic mitral valve disease were included. According to cardiac rhythm and duration of episode of AF, patients were divided into four groups: sinus rhythm group, paroxysmal AF group, persistent AF for less than 6 months group and persistent AF for more than 6 months group. Atrial tissue was obtained from the right atrial appendage, the right atrial free wall and the left atrial appendage respectively during open heart surgery. Total RNA was isolated and reversly transcribed into cDNA. In a semi - quantitative polymerase chain reaction the cDNA of interest and of glyceralde-hyde3 - phosphate dehydrogenase (GAPDH) were amplified and separated by ethidium bromide - stained gel electrophoresis. Multiple liner regress was used for correlation between the mRNA amount and age, sex, right atrial diameter (RAd) and left a     

Hypogonadism in a Patient with Two Novel Mutations of the Luteinizing Hormone β-Subunit Gene Expressed in a Compound Heterozygous Form

Context: LH gene mutations are rare; only four mutations have been described. The affected individuals are hypogonadal. Patient: We describe the clinical features of a 31-yr-old man who presented with delayed puberty and azoospermia and was found to have hypogonadism associated with an absence of circulating LH. Main Outcome Measures and Results: The patient had a 12-bp deletion in exon 2 in the LH β-subunit gene and a mutation of the 5' splice site IVS2+1G→T in the same gene present in a compound heterozygous state. The first mutation predicts a deletion of four leucines of the hydrophobic core of the signal peptide. The second mutation disrupts the splicing of mRNA, generating a gross abnormality in the processing. The patient's heterozygous parents were clinically normal. The phenotype of a 16-yr-old sister of the proband, carrying the same mutations, was characterized by normal pubertal development and oligomenorrhea. Conclusion: This report unravels two novel mutations of the LH gene critical for synthesis and activity of the LH molecule. The insight gained from the study is that normal pubertal maturation in women can occur in a state of LH deficiency, whereas LH is essential for maturation of Leydig cells and thus steroidogenesis, puberty, and spermatogenesis in man. These mutations should be considered in girls and boys with selective deficiency of LH.     

Polymorphism in the First Intron of Interferon-Gamma Gene (+874T→A) in Iranian Patients with Brucellosis

Background: Brucella is a gram-negative bacterium, causing acute and chronic infection in humans and animals. Cell-mediated immunity is the main protective immune response against Brucella spp. Activation of macrophages by IFN-γ and generation of reactive oxygen intermediates and nitric oxide are the main immunologic mechanisms responsible for control of Brucella infection. Objective: To investigate the correlation between IFN-γ gene polymorphism and brucellosis. Methods: 195 patients with brucellosis, 186 healthy patients' family members and 82 healthy farmers who kept infected animals and consumed their contaminated dairy products were selected to take part in the study. IFN-γ genotyping at position +874 (T→A) was carried out by allele specific polymerase chain reaction (AS-PCR) method. Results: The frequency of AT and TT genotypes significantly increased in farmers compared to patients with brucellosis (P=0.03) while there was no significant difference in genotype distribution between patients and their healthy family members. Conclusion: IFN-γ (+874) AA genotype is probably a genetic factor that contributes to the susceptibility of the individuals to brucellosis.     

Pattern of Acid Reflux in Patients with Reflux Esophagitis ‘Resistant’ to H2-Receptor Antagonists

Ambulatory 24-h esophageal pH monitoring was carried out in 54 patients with erosive/ulcerative reflux esophagitis before a 12- to 24-week treatment with either ranitidine, 150 to 300 mg twice daily, or famotidine, 20 to 40 mg twice daily. After this period, 21 patients continued to present endoscopic evidence of esophagitis. Patients who did not respond to the therapy showed a more severe pretreatment pattern of acid reflux than those who healed, with regard to both median percentage time of reflux (16.2% versus 11.0%, respectively, p < 0.05) and median number of reflux episodes (88.0 versus 55.0; p < 0.05). Ambulatory 24-h esophageal pH-metry is therefore to be recommended in all patients with acid reflux symptoms, even in those who already show endoscopic lesions of the esophageal mucosa, since this test is a valid prognostic indicator of response to treatment.