Effect of Helicobacter pylori infection and eradication therapy on interleukin-6 levels in patients with Familial Mediterranean Fever.

It is being questioned if Helicobacter pylori infection, which causes a chronic inflammatory response, can increase the frequency and severity of attacks in patients with Familial Mediterranean Fever (FMF) and if the impact of inflammatory response can be diminished by eradication of the infection. To evaluate if there is difference in interleukin (IL)-6 levels of H. pylori-positive and -negative patients both before and during FMF attacks; if there is a change in IL-6 levels following successful eradication treatment; and if MEFV gene mutations have an effect on IL-6 levels. IL-6 levels were evaluated in 47 FMF patients before and during FMF attacks. Genetic testing to determine M694V, M694I, E148Q, V726V, M680I mutations was also performed in all patients. IL-6 levels were also determined after successful eradication of the infection in H. pylori-positive patients. IL-6 levels were compared in H. pylori-positive and -negative patients, and before and after eradication treatment in patients who cleared the infection. Number of patients in tested mutation groups was not enough to compare IL-6 levels in these groups. Thirty-four patients (73.9%) were H. pylori-positive. Before FMF attack there was no statistically significant difference in IL-6 levels of H. pylori-positive and -negative groups. IL-6 levels were significantly higher in both groups during the attacks than before the attacks (p < 0.05). There was a statistically significant decline in IL-6 levels both before and during FMF attacks, following eradication therapy in patients who cleared the infection (p < 0.05). In patients with homozygous M694V mutation, IL-6 levels before and during the FMF attacks were not significantly different in H. pylori-positive and -negative groups, despite a much lower level found in H. pylori-negative group (p > 0.05). Comparisons were not performed in other mutation groups because of small number of patients in each group. C-reactive protein (CRP) and fibrinogen levels were not significantly different between the groups (p > 0.05). We believe that the observation of IL-6 levels are lower both before and during FMF attacks both in H. pylori-negative patients and in patients who cleared the infection after eradication therapy is very important in the determination of the role of eradication of H. pylori on decreasing the frequency and severity of FMF attacks. As for today, the correlation between H. pylori infection and FMF seems unlikely; however, studies evaluating the interaction of cytokines in both diseases and their relations and roles will be needed to reach better conclusions.

Diagnostic value of ischaemia-modified albumin for predicting myocardial ischaemia during myocardial perfusion scintigraphy.

During acute ischaemia the N-terminal site of albumin is altered, reducing its binding capacity; the modified protein is termed ischaemia-modified albumin (IMA). IMA is a sensitive marker of acute myocardial ischaemia but its diagnostic value in chronic angina pectoris patients is unclear. We investigated changes in blood levels of IMA during myocardial perfusion scintigraphy in patients with chronic angina pectoris in a study including 26 male and 20 female patients, with mean age 60 years. Technetium 99m perfusion imaging detected myocardial ischaemia in 26 patients. Coronary angiography was carried out in these 26 ischaemic patients. Mean IMA values at rest and peak exercise were significantly higher in the ischaemic than the nonischaemic group. All IMA values were considered negative for cardiac ischaemia. Peak exercise IMA was significantly lower than the pre-exercise level only in the non-ischaemic group. IMA measurements during myocardial perfusion scintigraphy are not helpful in the diagnosis of myocardial ischaemia in patients with stable angina pectoris.

Molecular characterisation of six patients with prolidase deficiency: identification of the first small duplication in the prolidase gene and of a mutation generating symptomatic and asymptomatic outcomes within the same family.

Prolidase deficiency (PD) is a rare autosomal recessive connective tissue disorder caused by mutations in the prolidase gene. The PD patients show a wide range of clinical outcomes characterised mainly by intractable skin ulcers, mental retardation and recurrent respiratory infections. Here we describe five different PEPD mutations in six European patients. We identified two new PEPD mutant alleles: a 13 bp duplication in exon 8, which is the first reported duplication in the prolidase gene and a point mutation resulting in a change in amino acid E412, a highly conserved residue among different species. The E412K substitution is responsible for the first reported phenotypic variability within a family with severe and asymptomatic outcomes.

Vitamin B12, folate, homocysteine and urinary methylmalonic acid levels in infants.

Serum vitamin B12 and folate, and their functional markers, plasma homocysteine and urinary methylmalonate (uMMA) were measured in 204 healthy, term infants at birth, and at 2 and 6 months. Compared with infants receiving formula food, those fed mother's milk had lower vitamin B12 and folate at 2 and 6 months. In infants receiving mother's milk, vitamin B12 levels were similar at birth (238 pg/ml) and 2 months (243 pg/ml), whereas with formula milk the level was significantly higher at 2 months (558 pg/ml) than at birth (257 pg/ml). Vitamin B12 was negatively correlated with homocysteine at birth and 6 months. The level of uMMA (mmol/mol creatinine) was higher at 2 (mother's milk, 25.5; formula, 23.97) and 6 months (19.77; 15) than at birth (11.97; 10.88), and was not correlated with vitamin B12 levels. Homocysteine may be a reliable marker of vitamin B12 status in neonates and infants; however, uMMA is not suitable as a marker of vitamin B12 status.

Co-existence of Helicobacter pylori infection in patients with Familial Mediterranean Fever (FMF) and the effect of Helicobacter pylori on the frequency and severity of FMF attacks.

BACKGROUND: The inflammatory reactions both in Familial Mediterranean Fever and in Helicobacter pylori infection have similarities. Whether there is interactions in case of co-existence of both diseases has not been evaluated. AIM.: To evaluate, if there is a significant relation between H. pylori infection and Familial Mediterranean Fever; if H. pylori has an effect on the frequency and severity of Familial Mediterranean Fever attacks; and if eradication treatment has any affects. METHODS: Thirty-two Familial Mediterranean Fever patients were tested for H. pylori infection. Acute phase responses were evaluated and attack frequency and severity were determined in both H. pylori-positive and H. pylori-negative groups. Same determinations were done after the eradication treatment in H. pylori-positive patients. Levels of acute phase determinants as well as frequency and severity of attacks were compared in H. pylori-positive and -negative groups. RESULTS: C-reactive protein, erythrocyte sedimentation rate, white blood count and fibrinogen levels were significantly (p<0.01) higher during the attacks than before the attacks in all patients. However, there was no difference between the groups. H. pylori-positive patients have a higher frequency and a longer duration of attacks when compared to H. pylori-negative patients before treatment (p<0.05). The frequency was also significantly lower and duration was shorter in patients whose infections were eradicated (p<0.05). CONCLUSION: H. pylori infection was not significantly frequent in our group of Familial Mediterranean Fever patients. H. pylori can decrease both the frequency and the duration of the attacks. Studies that will evaluate the relationship of H. pylori and MEFV gene along with the roles of yet unknown cytokines, which can presumably play a role in the pathogenesis of both diseases, are needed to reach better conclusions.

Transport of glutathione conjugate in erythrocytes from aged subjects and susceptibility to oxidative stress following inhibition of the glutathione S-conjugate pump.

The aim of the present study was to investigate the effect of donor aging on the glutathione conjugate transport in erythrocytes and whether it plays a role in the resistance to oxidative stress of the erythrocytes of aging subjects. In our comparative study on intact erythrocytes of healthy aging and young adults, in which 2,4-dinitrophenyl-S-glutathione (DNP-SG) was used as model glutathione S-conjugate, we found that the efflux of DNP-SG remained unchanged in the aged subjects. This result suggests that the detoxification function is maintained against the chemical stress employed in erythrocytes of aging subjects. In the assay conditions used, which were optimized to obtain maximal inhibition of glutathione S-conjugate transport, our results also indicated that the susceptibility of erythrocytes to in vitro lipid peroxidation generated by cumene hydroperoxide was enhanced by pretreatment with DNP-SG inhibitors in both age groups. However, the difference in susceptibility was not a function of aging. Further, the results suggested that inhibition of glutathione S-conjugate pump may impair cellular protection of the erythrocytes against oxidative damage.

Urine methylmalonic acid measurements for the assessment of cobalamin deficiency related to neuropsychiatric disorders.

BACKGROUND: Detection of cobalamin deficiency is clinically important for a better understanding of neuropsychiatric diseases, and why the deficiency occurs more frequently than previously anticipated. However, serum cobalamin measurements have a limited ability to diagnose a deficiency state. OBJECTIVE: To evaluate functional cobalamin status in neuropsychiatric patients using an appropriate photometric urine methylmalonic acid (MMA) determination method that could be easily adapted to all routine clinical laboratories. METHODS: We modified the old photometric method used for determining urinary MMA concentrations. MMA measurements were made in first morning urine samples with normalizing by creatinine concentrations. The serum cobalamin, total homocysteine (tHcy), folate, red cell folate, and urinary MMA concentrations taken from 17 psychosis, 28 depression, 16 dementia patients and 47 healthy people were analyzed using the ROC, correlation and multiple regression analysis. RESULTS: The modified method was found to have better recovery (96-103%) and CV% values than the old method. Mean +/- SDs of uMMA and cobalamin concentrations were 11.49 +/- 4.93 mmol/mol creatinine, and 231 +/- 151 pg/mL in psychosis and depression group, and 6.04 +/- 1.93 mmol/mol creatinine and 308 +/- 140 pg/mL in control group, respectively. Those in the dementia group were 11.53 +/- 4.0 mmol/mol creatinine and 231 +/- 84 pg/mL, and in the control group 6.05 +/- 1.94 mmol/mol creatinine and 364 +/- 188 pg/mL. There was a good correlation between urinary MMA and serum Vitamin B(12) determinations for all groups at a confidence level (p) of 99%. The correlation between urinary MMA and red cell folate was also significant at p = 95% for depression, psychosis and control groups, and p = 99% for dementia group. In the ROC analyses, area under the curve values for uMMA, B12 and tHcy were 0.842, 0.796 and 0.728, respectively. CONCLUSIONS: A sensitive and easy photometric method has been presented. When cobalamin deficiency is suspected in neuropsychiatric patients, photometric urinary MMA determination analysis can be the first diagnostic test used. If the urinary MMA concentration is above the reference value, serum cobalamin levels can be determined for further diagnosis.

Familial Mediterranean fever. A review of the disease and clinical and laboratory findings in 105 patients.

BACKGROUND: The diagnosis of familial Mediterranean fever still remains clinical, since no specific laboratory test exists, other than a molecular genetic test which is not widely available. AIM: To evaluate the clinical findings in 105 Turkish patients; to compare these findings with those in the literature; and to make a brief review of the disease. METHODS: A total of 105 familial Mediterranean fever patients were evaluated either retrospectively (for those diagnosed before 1997), or prospectively (for those after 1997). A diagnostic criteria set was used in addition to the clinical and laboratory findings that can be seen in familial Mediterranean fever, including the newly described manifestations. Previously selected clinical and laboratory parameters were observed for three consecutive days. RESULTS: Of our patients, 88.5% were of Turkish, 3.8% of Armenian and 7. 6% of Jewish origin. Family history was positive in 87 (82.8%) patients. Involved site was peritoneum in 97 (92%), joints in 45 (42.8%) and pleura in 19 (18%). Frequency of myalgia/arthralgia was 24.7%, and skin findings were observed in 16. 1% of patients. Splenomegaly, not related to amyloidosis, was present in 21 (20%) patients. Meningeal, retinal or ovarian/testicular involvement was not observed. CONCLUSION: Identification of familial Mediterranean fever gene has led to the application of a molecular genetic test for the diagnosis of Familial Mediterranean Fever. Until genetic methods become widely available, diagnosis will remain clinical. Thus, awareness of various clinical forms and of the correct usage of diagnostic criteria in various patient populations is important.

Are individuals with glutathione S-transferase GSTT1 null genotype more susceptible to in vitro oxidative damage?

Recent epidemiological studies proposed that glutathione S-transferase (GST) T1 null genotype was correlated with an increased susceptibility to diseases associated with oxidative stress, including cancer. A comparative study using erythrocytes from individuals with GSTT1 null genotype was carried out to determine how resistance to oxidative stress is affected by lack of this gene, and whether the GST status of a person is an important factor in risk toward oxidant chemicals. Malondialdehyde and carbonyl levels and fluorescence and chemiluminescence formation were used as biomarkers of oxidative stress in erythrocytes exposed in vitro to cumene hydroperoxide (CumOOH), an oxidizing agent. When peroxidation-dependent changes in these parameters were compared between GSTT1 null genotype and controls, who are both GSTM1 and GSTT1 positive, no significant differences were found between the two genotypes, although the erythrocytes of the GSTT1 null group had lower GSTT1 activity toward CumOOH. Our results indicate that erythrocytes from individuals with GSTT1 null genotype are not abnormally susceptible to CumOOH-induced oxidant challenge.

The presence of prolidase activity in amniotic fluid and its evaluation as a maturity test.

Prolidase (EC: 3.4.13.9) catalyses the hydrolysis of the peptide bond involving the imino nitrogen of proline or hydroxyproline. Because of the high proportion of imino acids in collagen, this enzyme plays an important role in its degradation. Since collagen turnover rate is expected to be high during fetal growth, the level of prolidase activity may reflect the degree of fetal maturation. In this study, amniotic fluid prolidase I activity was measured in term and preterm pregnancies. Lecithin concentration, which has been widely used for predicting fetal lung maturity, was also measured. Prolidase I activity was positively correlated with lecithin levels (n = 30; r = 0.42; p < 0.02), and also with birth weight of the babies (n = 30; r = 0.52; p < 0.01) in the term-mature group. Dysmature babies had significantly lower prolidase I activity in the amniotic fluid which was thought to be indicative of growth retardation.

The effect of hyperbaric oxygen treatment on the inflammatory changes caused by intraperitoneal meconium.

Exposure of the peritoneal cavity to meconium causes a marked inflammatory response. The effect of intraperitoneal meconium on intestinal morphology and plasma nitrite and nitrate (NO2(-) + NO3(-)) levels and how this inflammatory process is influenced by hyperbaric oxygen (HBO) treatment were investigated in this study. The purpose was to determine whether HBO treatment could be considered a useful adjunct in the resuscitative treatment of severely ill patients admitted with meconium peritonitis (MP). Rats were divided into three groups. Human meconium (MP group, n=10) and sterile saline (control group, n=10) were injected intraperitoneally for 3 days. The procedure for meconium injection was combined with HBO treatment for the HBO group (n=10). HBO was administered for 7 days. In all groups, peritoneal swap cultures, plasma NO2(-) + NO3(-) levels, intestinal diameters, and macroscopic and microscopic changes in the intestine were determined on the 8th day. Bacterial growth was not detected in the peritoneal swap cultures. There was a significant difference in NO2(-) + NO3(-) levels between the MP and HBO groups ( P<0.05), between the MP and control groups ( P<0.01), and between the HBO and control groups ( P<0.05). Thin fibrinous adhesions in both the MP and HBO groups, and thickened and dilated intestinal loops in the MP group were observed macroscopically. The intestinal diameter in the MP group was significantly greater than in the HBO and control groups. The only microscopic difference was seen in the serosal layer. Compared with the animals in the control and HBO groups, the intestine of the rats in the MP group showed prominent serosal thickening, edema, capillary proliferation and cellular infiltration. The ameliorated inflammatory changes and decreased dilatation of the intestine accompanied by a significant decrease in NO2(-) + NO3(-) levels suggest that as an adjunctive treatment, HBO may have a beneficial effect in the resuscitative treatment of meconium peritonitis.

The development and validation of a competitive, microtiter plate enzymeimmunoassay for human albumin in urine.

We have described a fast, simple and sensitive microtiter scale, solid phase, competitive enzymeimmunoassay (EIA) for the determination of urinary albumin. The albumin used in the test system was purified by the combination of PEG precipitation and DEAE-cellulose column chromatography. In this EIA, microtiter plates were coated with rabbit antihuman albumin IgG, and incubated with HRP-albumin conjugate with either sample or standards. O-phenylenediamine (OPD) and H2O2 solution was used as substrate for HRP. Results obtained correlate well (r = 0.994) with those of an in-house RIA in which same antibody and standards were used as in EIA. The present assay covers the range of 0.5 to 10 mg/L and can be performed in 2 hours. The detection limit was 0.15 mg/L of albumin. Within-assay coefficient of variation was 8.1% and 6.6% and between-assay variation was 10.6% and 8.6% at 1.25 and 2.5 mg/L respectively.

The effect of Helicobacter pylori eradication on gastric juice and blood ammonia concentrations and on visual evoked potentials in cirrhotics.

BACKGROUND: The primary source of ammonia is the gut. Ammonia can also be generated by the urease activity of Helicobacter pylori in the gastric mucosa. The aim of this study was to investigate the effect of H. pylori eradication on blood and gastric juice ammonia levels and on visual evoked potential (VEP) recordings in cirrhotic patients. MATERIALS AND METHODS: Male patients with cirrhosis and H. pylori infection were prospectively evaluated. All patients were given triple regimen for eradication for 10 days. Gastroscopy together with gastric juice sampling for ammonia and mucosal sampling for H. pylori status was performed before and after therapy. Gastric juice and blood ammonia levels were measured and VEP recordings were obtained before and after treatment. RESULTS: Twenty-seven patients were included in the study. Patients with overt clinical hepatic encephalopathy were excluded from the study. Twenty-four out of twenty-seven patients became H. pylori-negative after the treatment. Ammonia measurements and VEP recordings were evaluated in the 24 patients in whom eradication was successful. A slight but statistically significant decrease in blood and a considerable decrease in gastric juice ammonia levels were observed after treatment [from 44.23 micromol/l to 41.6 micromol/l compared with 3234 micromol/l to 2709 micromol/l, respectively (p <.05)] in patients in whom H. pylori was eradicated. VEP recordings were abnormal in 14 out of 24 patients before the treatment. Only four of these 14 patients with abnormal recordings showed improvement. (p > .05). CONCLUSIONS: Helicobacter pylori eradication in cirrhotics decreases blood and gastric juice ammonia concentrations whereas it does not provide an improvement in VEP recordings.

CagA status in dyspeptic patients with and without peptic ulcer disease in Turkey: association with histopathologic findings.

BACKGROUND: CagA seropositivity is closely associated with that of vacuolating cytotoxin (VacA). Helicobacter pylori strains positive for both VacA and CagA were reported to be strongly associated with peptic ulcer disease. Different results reporting that cagA gene is not associated with more serious diseases, lowers the importance of CagA protein as a marker. In this study, CagA seropositivity is examined in Turkish peptic ulcer and nonulcer dyspepsia patients; histopathologic scores of CagA (+) and CagA (-) groups were compared. MATERIALS AND METHODS: Sixty consecutive patients (one gastric ulcer, 13 duodenal ulcer and 46 nonulcer dyspepsia) (mean age 40.9 +/- 14.7; 33 women, 27 men) with dyspeptic complaints who underwent upper gastrointestinal (GI) endoscopy were included. Biopsies from the antrum and corpus were used for histopathologic examination and for rapid urease test. H. pylori-negative patients comprised the control group. Histopathologic findings were graded using a previously described grading system (for inflammation, activity, atrophy, intestinal metaplasia and H. pylori, grades from 0 to 3 were used to quantify the findings). In H. pylori-positive patients, antibodies against CagA protein were determined using an ELISA METHOD: RESULTS: H. pylori was (+) in 46 patients. One duodenal ulcer and 13 nonulcer dyspepsia patients were negative for H. pylori. CagA positivity is significantly higher in peptic ulcer patients [12/12] than in nonulcer dyspepsia patients [25/33]. While inflammation, activity and atrophy scores were significantly higher in CagA positive patients, intestinal metaplasia and H. pylori load scores were not. Although the histopathologic scores in controls were lower than CagA (-) group, statistical significance was observed only in inflammation and intestinal metaplasia scores. CONCLUSION: Development of more prominent gastritis and severe atrophy in CagA (+) patients is an indicator of the importance of CagA rather than H. pylori load. Therefore, we suggest that nonulcer dyspepsia patients should also be tested for CagA status along with the tests for H. pylori status; and a positive CagA testing should be considered as an indication for eradication treatment. If CagA is negative, further assesment should be performed to decide whether or not to treat the patient.

Effect of partial oxygen supply on mitochondrial electron transport system during complete cardiac ischemia.

During complete ischemia we assessed myocardial utilization of the small amount of oxygen available. We also determined whether blood cardioplegia has any advantage over crystalloid cardioplegia in this setting. Patients with preserved left ventricular myocardial function and without anterolateral wall infarct or aneurysm were included to the study. Intermittent cold blood and crystalloid cardioplegia were used in 10 patients (group BC) and 9 patients (group CC), respectively. From myocardial biopsies, obtained before and after ischemia, complete electron transport system (ETS) enzyme activities (NDH, SDH, NCCR, SCCR, and COX) and lactate content were analyzed. Biochemical and hemodynamic analyses also were done. Myocardial and blood temperatures were monitored. Ischemic time was longer in group CC (p < 0.05). There were no important differences in biochemical and hemodynamic variables between the two groups. In addition, there was no difference in NDH and SDH activities as well as COX/SCCR and COX/RS-NCCR ratios between the two groups before and after ischemia. After Ischemia, RS-NCCR in group CC and SCCR and COX activities in both groups were lower than the control. For all enzymes, activity change ratios were not different between groups. Myocardial lactate content was increased in both groups after ischemia. However, the increase in group BC was less (p < 0.01). Based on our findings, we believe that the superiority of blood cardioplegia over crystalloid cardioplegia does not depend on oxygen content, but on other factors such as buffering and free oxygen radical scavenger effects among others. However, with the warm and continuous blood cardioplegia technique, oxygen content might be more important.