5-aminosalicylic acid inhibits the expression of oncomiRs and pro-inflammatory microRNAs: an in vitro study.

5-aminosalicylic acid (5-ASA) is commonly used as the first-line treatment for ulcerative colitis (UC). In this study, we show that the mechanism responsible for the protective effect of 5-ASA is associated with the modulation of non-coding microRNA molecule (miRNA) expression. Stimulation of human intestinal epithelial cells (Caco-2) with 1000 µM of 5-ASA suppressed the levels of miR-125b, miR-150, miR-155, miR-346 and miR-506, which are known to be involved in the regulation of colitis and/or colorectal cancer in patients with inflammatory bowel disease. The 5-ASA-induced inhibitions of these miRNAs were associated with significant inductions of their target genes such as vitamin D receptor (VDR), suppressor of cytokine signaling (SOCS1), Forkhead box O (FOXO3a) and DNA methyltransferase 1 (DNMT1). The relationships between the selected miRNAs and their target genes were further confirmed in Caco-2 cells transfected of with specific miRNA inhibitors or miRNA mimics. Moreover, we showed that 5-ASA has the potential to hinder miR-155 expression induced by the transfection of miR-155 mimic into Caco-2 cells. These findings underline the anti-inflammatory and chemoprotective effects of 5-ASA treatment.

Clinical and diagnostic approach in unsolved CDG patients with a type 2 transferrin pattern.

Dysmorphic features, multisystem disease, and central nervous system involvement are common symptoms in congenital disorders of glycosylation, including several recently discovered Golgi-related glycosylation defects. In search for discriminative features, we assessed eleven children suspected with a Golgi-related inborn error of glycosylation. We evaluated all genetically unsolved patients, diagnosed with a type 2 transferrin isofocusing pattern in the period of 1999-2009. By combining biochemical results with characteristic clinical symptoms, we used a diagnostic flow chart to approach the underlying defect in patients with congenital disorders of glycosylation-IIx. According to specific symptoms and laboratory results, we initiated additional, targeted biochemical and genetic studies. We found a distinctive spectrum of congenital disorders of glycosylation type 2-associated anomalies including sudden hearing loss, brain malformations, wrinkled skin, and epilepsy in combination with skeletal dysplasia, dilated cardiomyopathy, sudden cardiac arrest, abnormal copper and iron metabolism, and endocrine abnormalities in our patients. One patient with severe cortical malformations and mild skin abnormalities was diagnosed with a known genetic syndrome, due to an ATP6V0A2 defect. Here, we present unique congenital disorders of glycosylation type 2-associated anomalies, including both ATPase-related and unrelated cutis laxa and sensorineural hearing loss, a recently recognized symptom of congenital disorders of glycosylation. Based on our findings, we recommend clinicians to consider congenital disorders of glycosylation in patients with cardiac rhythm disorders, spondylodysplasia and biochemical abnormalities of the copper and iron metabolism even in absence of intellectual disability.

Ophthalmological abnormalities in children with congenital disorders of glycosylation type I.

BACKGROUND: Children with congenital disorders of glycosylation (CDG) type Ia frequently present with ocular involvement and visual loss. Little is known, however, about the occurrence of ophthalmological abnormalities in other subtypes of CDG syndrome. METHODS: We evaluated 45 children sequentially diagnosed with CDG type I for the presence of ocular abnormalities at the time of the diagnosis and during follow-up. We compared the various ophthalmic findings in the different CDG subgroups. RESULTS: Of the 45 patients, 22 had CDG type Ia, nine had CDG type Ic and 14 had a so-far undiagnosed biochemical background (CDG type Ix). We found ocular anomalies in 28 of the 45 children. Three had unique findings, including congenital cataract, retinal coloboma and glaucoma. A few CDG type Ia patients showed a sequential occurrence of symptoms, including retinitis pigmentosa or cataract. CONCLUSIONS: Ophthalmic findings are frequent in CDG syndrome involving both the anterior and posterior segment of the eye. The disorder might lead to abnormal development of the lens or the retina, cause diminished vision, alter ocular motility and intraocular pressure. We suggest routine screening and follow-up for ophthalmological anomalies in all children diagnosed with CDG syndrome to provide early treatment and adequate counselling.

Congenital disorder of glycosylation type Ix: review of clinical spectrum and diagnostic steps.

Congenital disorder of glycosylation type I (CDG I) represent a rapidly growing group of inherited multisystem disorders with 13 genetically established subtypes (CDG Ia to CDG Im), and a high number of biochemically unresolved cases (CDG Ix). Further diagnostic effort and prognosis counselling are very challenging in these children. In the current study, we reviewed the clinical records of 10 CDG Ix patients and compared the data with 13 CDG Ix patients published in the literature in search for specific symptoms to create clinical subgroups. The most frequent findings were rather nonspecific, including developmental delay and axial hypotonia. Several features were found that are uncommon in CDG syndrome, such as elevated creatine kinase or arthrogryposis. Distinct ophthalmological abnormalities were observed including optic nerve atrophy, cataract and glaucoma. Two subgroups could be established: one with a pure neurological presentation and the other with a neurological-multivisceral form. The first group had a significantly better prognosis. The unique presentation of microcephaly, seizures, ascites, hepatomegaly, nephrotic syndrome and severe developmental delay was observed in one child diagnosed with CDG Ik. Establishing clinical subgroups and increasing the number of patients within the subgroups may lead the way towards the genetic defect in children with a so far unsolved type of the congenital disorders of glycosylation. Raising awareness for less common, non-CDG specific clinical features such as congenital joint contractures, movement disorders or ophthalmological anomalies will encourage clinicians to think of CDG in its more unusual presentation. Clinical grouping also helps to determine the prognosis and provide better counselling for the families.

Defining the phenotype in an autosomal recessive cutis laxa syndrome with a combined congenital defect of glycosylation.

Autosomal recessive cutis laxa is a genetically heterogeneous condition. Its molecular basis is largely unknown. Recently, a combined disorder of N- and O-linked glycosylation was described in children with congenital cutis laxa in association with severe central nervous system involvement, brain migration defects, seizures and hearing loss. We report on seven additional patients with similar clinical features in combination with congenital disorder of glycosylation type IIx. On the basis of phenotype in 10 patients, we define an autosomal recessive cutis laxa syndrome. The patients have a complex phenotype of neonatal cutis laxa, transient feeding intolerance, late closure of the fontanel, characteristic facial features including down-slanting palpebral fissures, short nose and small mouth, and developmental delay. There is a variable degree of the central nervous system involvement and variable systemic presentation. The biochemical analysis using transferrin isoelectric focusing gives false negative results in some of the youngest patients. Analysis of the apolipoprotein C-III isoelectric focusing, however, is diagnostic in all cases.

Transferrin hypoglycosylation in hereditary fructose intolerance: using the clues and avoiding the pitfalls.

Hereditary fructose intolerance (HFI) is caused by a deficiency of aldolase B due to mutations of the ALDOB gene. The disease poses diagnostic problems because of unspecific clinical manifestations. We report three cases of HFI all of whom had a chronic disease with neurological, nephrological or gastroenterological symptoms, whereas nutritional fructose intolerance, the pathognomonic sign of HFI, was apparent only in retrospect. In all patients a hypoglycosylated pattern of transferrin isoforms was found but was misinterpreted as a sign of CDG Ix. The correct diagnosis was achieved with marked delay (26, 36 and 24 months, respectively) by sequencing of the ALDOB gene two common mutations were identified on both alleles or on one (A150P/A175D, A150P/-, and A150P/A175D). The diagnosis was further supported by normalization of transferrin isoforms on a fructose-free diet. Data available in two patients showed that following the fructose restriction the type I pattern of carbohydrate-deficient transferrin detectable on fructose-containing diet disappeared after 3-4 weeks. These cases illustrate that in the first years of life HFI may show misleading variability in clinical presentation and that protein glycosylation analysis such as transferrin isofocusing may give important diagnostic clues. However, care should be taken not to misinterpret the abnormal results as CDG Ix as well as to remember that a normal profile does not exclude HFI due to the possibility of spontaneous fructose restriction in the diet. The presented data also emphasize the usefulness of ALDOB mutation screening for diagnosis of HFI.

Increased plasma glutathione peroxidase activity in patients with acute myocardial infarction.

BACKGROUND: Aim of the study was to determine the concentration of selenium (Se) and the activity of glutathione peroxidase (GSH-Px) in patients with acute myocardial infarction (AMI) and to observe the behavior of these parameters during thrombolysis therapy. MATERIAL AND METHODS: The study comprised two groups of AMI patients and a control group. The first group consisted of 49 patients from whom blood samples were taken after admission to the intensive care unit and subsequently after 3, 7, 14 and 30 days of hospitalization. In the second group of patients (n = 18) blood was taken for measuring only the GSH-Px activity in plasma. In this group blood samples were collected after admission to the hospital, 6, 12, 24, 48 hours, 3, 7, 14 and 30 days later. Control group comprised of 58 healthy subjects. Se levels in whole blood and plasma were measured spectrofluorometrically with 2,3-diaminonaphthalene as a complexing reagent. GSH-Px activity in red cells and plasma was measured spectrofluorometrically with t-butyl hydroperoxide as substrate. RESULTS: In the first group of patients Se concentrations in whole blood and plasma as well as GSH-Px activities in red cells and plasma did not differ significantly from healthy subjects. Both Se levels and GSH-Px activities were stable during the entire period of the study. In the second group of patients, however, plasma GSH-Px activity increased after admission and reached the highest value after 48 hours. This activity was significantly higher compared to healthy subjects (p < 0.004) and to the mean initial activity of this group (p < 0.02). In the later period the activity decreased to the values of healthy subjects. CONCLUSION: We suggest that the increased activity of GSH-Px in plasma of AMI patients is the response of the organism to the increased levels of reactive oxygen species produced during reperfusion and thrombolysis.

Genomic organization of the human phosphomannose isomerase (MPI) gene and mutation analysis in patients with congenital disorders of glycosylation type Ib (CDG-Ib).

CDG-Ib is the "gastro-intestinal" type of the congenital disorders of glycosylation (CDG) and a potentially treatable disorder. It has been described in patients presenting with congenital hepatic fibrosis and protein losing enteropathy. The symptoms result from hypoglycosylation of serum- and other glycoproteins. CDG-Ib is caused by a deficiency of mannose-6-phosphate isomerase (synonym: phosphomannose isomerase, EC 5.3.1.8), due to mutations in the MPI gene. We determined the genomic structure of the MPI gene in order to simplify mutation detection. The gene is composed of 8 exons and spans only 5 kb. Eight (7 novel) different mutations were found in seven patients with a confirmed phosphomannose isomerase deficiency, analyzed in the context of this study: six missense mutations, a splice mutation and one insertion. In the last, the mutation resulted in an unstable transcript, and was hardly detectable at the mRNA level. This emphasizes the importance of mutation analysis at the genomic DNA level.

[Does any relationship exist between metabolic disturbances and some markers of renal damage in patients with untreated essential hypertension?]. / Czy istnieja zaleznosci pomiedzy niektórymi zaburzeniami metabolicznymi a wybranymi wskaznikami uszkodzenia nerek u chorych z nieleczonym pierwotnym nadcisnieniem tetniczym?

The essential arterial hypertension is the second (after diabetes mellitus) cause of chronic renal failure which means a great social and economic burden to the society. It is well known that hypertension is a metabolic syndrome resulting in tissue injury. We tried to investigate the possible influence of some metabolic disturbances on renal function in nontreated essential hypertension. We have compared 25 patients with nontreated essential hypertension (11 women, 14 men) with 14 healthy volunteers (7 women, 7 men) matched for age. The patients' group was characterized by significantly higher urine excretion of NAG (N-acetyl-beta-D-glucosaminidase) (2.75 +/- 1.69 vs 1.82 +/- 1.46 p < 0.05) and a tendency to significantly higher urine fractional sodium excretion without significant difference in albumin excretion. These findings suggest that the tubular damage is present. We noticed the negative linear correlation between mean arterial pressure and (MAP) and NAG urine excretion in the group of hypertensive patients which may reflect the renal ischemia in tubulo-interstitial pathology. Our data suggests that in nontreated arterial hypertension the renal blood flow disturbances are the important cause of the deterioration of tubular function (which are earlier to glomerular damage).

A systematic analysis of secondary DNA transfer.

The Nature letter by R. van Oorschot and M. Jones (1) addressed two topics: the primary transfer of DNA from person to person or to various objects, and the secondary transfer of DNA through an intermediary. Forensic scientists have described the primary transfer of DNA and other biological evidence for many years. However, the authors also reported detecting secondary transfer of DNA from an object to a person's hands, which could adversely affect DNA typing in the forensic context. The prospect of secondary transfer raises questions of interest to both the legal and forensic communities. Therefore, we sought to evaluate parameters potentially leading to secondary DNA transfer. Our data do not support the conclusion that secondary transfer will compromise DNA typing results under typical forensic conditions.

A systematic analysis of PCR contamination.

In light of the strict legal scrutiny surrounding DNA typing at this time, it has become necessary to systematically address the issue of PCR contamination. To precisely define the parameters affecting PCR contamination under casework analysis conditions, PCR amplification reactions were intentionally compromised by employing sub-standard laboratory technique and by introducing secondary sources of DNA. The PCR parameters considered for potential sources of contamination include amplification set-up, amplification product handling, aerosol DNA and storage. In addition, analyst technique was evaluated by modifying or eliminating standard safeguards. Under the circumstances normally encountered during casework analysis, PCR contamination was never noted. Significantly, using the dot blot detection method, contamination was never observed when nanogram quantities of genomic DNA were mishandled or aerosolized. Contamination occurred only when amplification product was carelessly manipulated or purposefully sprayed near or directly into open tubes containing water or genomic DNA. Although standard precautions should be employed during PCR-based DNA typing, our data indicates that contamination during amplification procedures is not prevalent when detected by dot blot analysis.

Histological examination of the topography of the atrioventricular nodal artery within the triangle of Koch.

UNLABELLED: The treatment of choice in patients with drug-resistant atrioventricular nodal reentry tachycardia is radiofrequency fast or slow pathway ablation. Ablation of the reentrant circuit in the region of the His bundle, when approached from the anterior-superior region (fast pathway); can result in complete AV block. This is less likely if the posterior-inferior (in the region of coronary sinus ostium) approach is used (slow pathway ablation). The possibility that radiofrequency energy may damage the vascular supply to the AV node must be considered. In order to confirm this hypothesis observation was conducted on the autopsy material of 50 human hearts (20 F, 30 M) from 18 to 81 years of age. Specimens were taken containing the triangle of Koch (the apex- right fibrous trigone, the base- coronary sinus ostium). These histological blocks were sectioned in the frontal plane and stained using Masson's method. Koch's triangle was divided in the sagittal plane into 3 parts: inferior (between the base and the attachment of the tricuspid valve), central (between the base and the apex of the right fibrous trigone) and superior (between this trigone and the tendon of Todaro). It was observed that the AVN artery at the coronary sinus ostium level (the base of the triangle of Koch) was positioned in 68% in the central and in 32% in the inferior part of Koch's triangle. The AVN artery in the central part was removed from the endocardium 1 mm (18%), 2 mm (42%), 3 mm (22%), 4 mm (18%). In the inferior part 1 mm (26%), 2 mm (37%), 3 mm (37%). No statistically significant relationship was observed between those groups. CONCLUSIONS: 1) in 20% of examined hearts the AVN artery lay just beneath the endocardium near the coronary sinus ostium 2) there is a risk of the AVN artery coagulation during radiofrequency ablation in the slow pathway region.

Reduction in treatment delay by paramedic ECG diagnosis of myocardial infarction with direct CCU admission.

OBJECTIVES: To establish the feasibility of training paramedics of diagnose acute myocardial infarction by ECG before hospital admission and whether direct paramedic coronary care admission, arranged by very high frequency (VHF) radio communication with the coronary care unit (CCU), would reduce delay of thrombolysis treatment. DESIGN: Prospective controlled study. SETTING: District general hospital CCU and a local district ambulance paramedic service. PATIENTS: 124 patients with ECG evidence of myocardial infarction or ischaemia admitted directly to the CCU by the paramedic service were compared with 123 patients admitted by the emergency department and subsequently transferred to the CCU. MAIN OUTCOME MEASURES: ECG diagnostic accuracy by paramedics, and interval durations for CCU admission and thrombolysis. RESULTS: ECG diagnostic accuracy by the paramedics was 87.5% in the training phase and 92% in admission. The total call to thrombolysis interval was reduced from 154 to 93 minutes and the "door to needle" interval was reduced from 97 to 37 minutes. CONCLUSIONS: Trained paramedics can reliably diagnose myocardial infarction by ECG. The use of a direct admission procedure, by a VHF radio link to the CCU, substantially reduces the time interval for thrombolytic treatment after acute myocardial infarction.

Inhibition of phosphomannose isomerase by fructose 1-phosphate: an explanation for defective N-glycosylation in hereditary fructose intolerance.

Isoelectrofocusing of serum sialotransferrins from patients with untreated hereditary fructose intolerance (HFI) shows a cathodal shift similar to that in carbohydrate-deficient glycoprotein (CDG) syndrome type I and in untreated galactosemia. This report is on serum lysosomal enzyme abnormalities in untreated HFI that are identical to those found in CDG syndrome type I but different from those in untreated galactosemia. CDG syndrome type I is due to phosphomannomutase deficiency, a defect in the early glycosylation pathway. It was found that fructose 1-phosphate is a potent competitive inhibitor (Ki congruent to 40 microM) of phosphomannose isomerase (EC 5.3.1.8), the first enzyme of the N-glycosylation pathway thus explaining the N-glycosylation disturbances in HFI.

[Congestive heart failure and ventricular arrhythmia in relation to magnesium, potassium and sodium in serum and erythrocytes]. / Niewydolnosc krazenia i zaburzenia rytmu serca a magnez, potas i sód w osoczu i erytrocytach.

The aim of this study was to determine magnesium, potassium and sodium in plasma and erythrocytes in patients (14 male, 12 female; the average age 60.3) with congestive heart failure (CHF) (NYHA class II/III, III, IV) who were treated with digitalis, diuretics and vasodilators. All patients were subjected to 24 h ECG monitoring and ventricular arrhythmias classified according to LOWN, were analyzed in relation to electrolyte contents in erythrocytes and serum. Control group consisted of 20 persons (17 male; 3 female: the average age 58.7 years). Plasma and erythrocyte K and Na were determined by flame spectrophotometer; Mg was assayed by atomic absorption spectrophotometer. Statistical analysis was made by t-Student test. Results obtained suggest that patients with CHF require supplementation K+ with Mg2+ (tab. I). Our study has not revealed expected interrelationships between ventricular arrhythmias and electrolyte disturbances in patients with CHF. It should be stressed, however, that the small numbers of patients in the subgroups in our study might have blurred the possible relationship.

[Hypertension in pregnancy with special reference to its treatment]. / Nadcisnienie tetnicze u ciezarnych ze szczególnym uwzglednieniem leczenia.

Definition and classification of the arterial hypertension in pregnancy are discussed. An emphasis is on the problems of differential diagnosis between pre-eclampsia and other forms of hypertension. Use of hypotensive drugs in pregnant patients with particular reference to emergencies is also discussed. The treatment of pregnant women with hypertension is still a problem which require close co-operation of both an obstetrician and internist. Follow-up after labour is GP duty to find out if the patient remains hypertensive. If so, etiology of the disease should be again searched.