[Changing clinical image of celiac sprue in childhood]. / Menící se klinický obraz celiakální sprue v detském veku.

BACKGROUND: Celiac sprue is considered to be the second most common chronic disease in childhood after allergic diseases. At present, the prevalence of this disease is stated as high as approximately 1% in inhabitants of the North America and Western Europe. Aetiology of celiac sprue is multifactorial as it is in other chronic diseases. Pathogenetically, it is an autoimmune disease whose main autoantigene is the tissue transglutaminase. It affects those individuals carrying HLA-DO2 or HLA-DO8 gene and those who were exposed to wheat gliadine or similar amino acids (prolamines) in rye and barley. The purpose of the present study was to find out whether clinical manifestation of celiac sprue changed in our group of patients in course of 23 years. METHODS AND RESULTS: In forty-eight children celiac sprue was diagnosed according to histopathological and histochemical findings in the small intestine mucosa. In the children examined within 5 years in 1982-1987, main clinical symptom of celiac sprue was diarrhoea. In the control group of children examined within 7 months in 2004-2005, intestinal and extraintestinal symptoms in celiac sprue were equally distributed; the so-called sleeping forms occurred too. CONCLUSIONS: It is discussed what are the trigger mechanisms and the possible danger of celiac sprue manifestations and why celiac sprue is diagnosed in the older children at present time compared with the past years.

Analysis of facial skeleton shape in patients with complete unilateral cleft lip and palate: geometric morphometry.

The study is based on a longitudinal cephalometric follow-up of X-ray films of patients with a complete unilateral cleft lip and palate during puberty. Carthesian x, y coordinates of the total of 22 landmarks on the lateral films of 25 patients with the same diagnosis and therapy were established, which served as outcome data for shape analysis. The changes in facial shape experienced by patients between their 10th and 15th year were evaluated by the thin-plate splines (TPS) method. We used the TPS method to find a function which transforms the shape of the face at the age of 10 as well as the square TPS grid into the facial shape at the age of 15 years with the deformed TPS grid. With the help of Geometric PCA for Bookstein's coordinates we found individuals with different shapes who develop abnormally during puberty and whose development is not well predictable. During puberty the face becomes relatively elongated. The most pronounced deformation is caused by the shift of the first permanent molar anteriorly, or a relative shortening of the frontal part of the dental arch. In the anterior direction the position of landmarks changed only in the area of nose; the alveolar process shows retrusion. The position of the mandible in relation to the maxilla change mainly in the inferior direction; the mandibular angle shifts posteriorly. The shape of skeletal profile was compared with a group of patients with the same diagnosis who underwent different therapy.

[Helicobacter pylori infection in a group of symptomatic and asymptomatic children and adolescents in the Czech Republic]. / Infekce Helicobacter pylori u souboru symptomatických a asymptomatických detí a dospívajících v Ceské republice.

BACKGROUND: There exist only inconsistent results of studies on the relationship between Helicobacter pylori (H. pylori) infection and functional dyspepsia and/or recurrent abdominal pain in children and adolescents. The answer could bring a comparison of the epidemiological features of H. pylori infection between children and adolescents with dyspepsia and/or recurrent abdominal pain (symptomatic) and without these symptoms (asymptomatic), living in the Czech republic. METHODS AND RESULTS: Study includes children and adolescents (2-18 years) with upper gastrointestinal symptoms visiting paediatric gastroenterology department between 1994 and 1999. Age, sex, socioeconomic level matched control (asymptomatic) group consisted of children and adolescents visiting the general paediatric service with symptoms not related to the upper gastrointestinal tract. Demographic and socioeconomic factors, including parent(s) educational level, place of residence, living conditions, type of drinking water and pets in their household were evaluated. Serum of the children and adolescents was tested for H. pylori IgG by enzyme linked immunosorbent assay. In addition, the symptomatic children and adolescents underwent endoscopic examination and biopsies from gastric antrum and corpus were taken for the quick urease test, histology/histoscopy, and H. pylori cultivation. H. pylori infection in this group was stated when at least two tests were positive. Altogether 829 children and adolescents were examined, 624 cases were symptomatic and 205 represented controls (asymptomatic). The prevalence of infection was 33% among symptomatic children vs. 7.5% among controls (OR = 6.2, p.001) and was similar among boys and girls (32% vs. 34.5%, respectively). H. pylori prevalence increased with age among symptomatic children (10% for children below 6 years and 37% between 11 to 16 years) (p.001). In contrary, prevalence tended to fall with age among asymptomatic children (11% in children below 6 years vs. 6% in children over 10 years (OR = 2.0, 95% CI = 0.7-6.2). In both groups, an inverse correlation between H. pylori occurrence and mother's educational level was observed. The H. pylori incidence was 10% among asymptomatic children who drank water from municipal water supply or from a well vs. 3% among those who drank bottled water (OR = 4, 95% CI = 1.1 to 18, p.05). CONCLUSIONS: H. pylori infection was more prevalent among symptomatic children and adolescents vs. asymptomatic children and adolescents within the same population. H. pylori incidence increased with age among symptomatic children and adolescents and tended to fall among controls, showing deep differences between the two groups. With exception of the bottled water drinking, presence of any other factor studied represented significant risk for acquiring the H. pylori infection.

A 5-year randomized controlled, double-blind study of glycosaminoglycan polysulphuric acid complex (Rumalon) as a structure modifying therapy in osteoarthritis of the hip and knee.

OBJECTIVE: To determine the structure (disease) modifying effect of a glycosaminoglycan polypeptide association complex (GP-C; Rumalon) in patients with knee and hip osteoarthritis (OA). METHODS: Double-blind, randomized, placebo-controlled five-year study. Primary assessment criterion was change in radiographic joint space width between baseline and follow-up at 5 years. Secondary outcome criteria included Lequesne algofunctional index (LAI), pain on passive motion and consumption of non-steroidal antiinflammatory drugs (NSAIDs). The patients received 10 courses of injections of placebo or GP-C 2 ml intramuscularly in 5 years (two courses each year). Each course included 15 injections administered twice weekly. RESULTS: There were 277 patients with knee OA and 117 patients with hip OA. Control and GP-C treated groups were comparable as to sex, age, duration of disease, body weight, X-ray stage and value of LAI at the baseline. Knee joint space at 5 years decreased 0.37+/-0.08 (mean+/-standard deviation) mm for GP-C and 0.42+/-0.08 mm for placebo groups (P=0.68). Hip joint space at 5 years decreased 0.21+/-0.08 mm for GP-C and 0.22+/-0.08 mm for placebo groups (P=0.53). In a subset of patients with hip OA, Kellgren-Lawrence> or =2 and JSW> or =1 mm, there was a trend in favor of GPC for lower joint space narrowing in 5 years (P=0.11). In addition, there were no statistical differences between the treatment groups in LAI, pain on passive motion and consumption of NSAIDs. Side-effects after GP-C (14.5%) were rare, mild and not more frequent than in the placebo group (15%). CONCLUSION: We were not able to demonstrate a structure modifying effect of GP-C in OA of the hip or knee. Radiographic progression of OA in both knee and hip OA was lower than expected in both study groups.

Preferential silver reaction of nucleolar regions adjacent to fibrillar centers in ring shaped nucleoli of leukemic lymphocytes.

Silver stained proteins (SSPs) characteristic for interphasic nucleolus organizer regions (NORs) associated with fibrillar centers (FCs) and adjacent nucleolar regions of ring shaped nucleoli in leukemic lymphocytes exhibit a different sensitivity to the mild acid extraction including that with HCl. Such extractions permit a preferential visualization of fibrillar centers adjacent regions (FCARs) which are believed to represent sites of the ribosomal RNA (rRNA) transcription. The resistance of SSPs in FCARs to the extraction with HCl seems to be due to their binding to other components present in these regions. The extractibility of SSPs with HCl was influenced by the fixatives used. The largest resistance of SSPs to the extraction with HCl was noted after fixation with glutaraldehyde. In contrast, the largest extractibility of these proteins was observed after fixation with unbuffered formaldehyde.

[The effect of spermatozoal ultrastructure on the outcome of fertilization in vitro]. / Vliv ultrastruktury spermií na výsledek fertilizace in vitro.

UNLABELLED: The ultrastructure of sperm cells in cumulus oophorus of the human oocyte 20 hrs after in vitro fertilization (IVF) was studied. MATERIALS AND METHODS: Mechanically removed cumular cells were processed for electron microscopy and the spermatozoa deposited in the intracellular spaces as well as those penetrating into follicular cells were evaluated. The samples were divided into three groups. Group I contained cumuli from unfertilized oocytes, group II involved those from fertilized oocytes, but not pregnancy after embryo transfer. Group III was formed by cumuli from fertilized oocytes of patients, where pregnancy had been achieved. RESULTS: In group I numerous anomalies of spermatozoal ultrastructure were found, such as head malformations, incompleted chromatin condensation, large cytoplasmatic droplets, etc. In groups II and III, about 70% normal sperm cells were found. CONCLUSIONS: These findings provide evidence of the important role of the morphological quality of spermatozoa in the failure of in vitro fertilization, also in cases where anomalies could not be detected by the usual methods of sperm examination.

[Ultrastructural findings in the gastric mucosa in children and adolescents with chronic Helicobacter pylori-positive gastritis]. / Ultrastrukturální nálezy v zaludecní sliznici detí a mladistvých s chronickou Helicobacter pylori pozitivní gastritidou.

Gastric mucosa ultrastructure was studied in 32 children and adolescents with chronic superficial active and inactive gastritis. Helicobacter pylori was found most frequently free in mucous layer without direct contact with the mucosa. But in 7 children (21.9%), it occurred quite close to superficial epithelial cells and influenced their outlook. In two cases, it was registered in intracytoplasmic channels of intact parietal cells. Another two children with massive helicobacterial infection showed superficial epithelial cells with big mucus vacuoles and sequestration of apical cytoplasm.

[Helicobacter pylori infections: familial clustering and transmission]. / Infekce Helicobacter pylori: shlukování a prenos v rodinách.

BACKGROUND: The importance of Helicobacter pylori for the development of gastritis and gastro-duodenal ulceration is generally accepted. The objective of the presented work was to find out whether its transmission is involved in families of children examined on account of upper dyspeptic syndrome. Aggregation and transmission of Helicobacter pylori infection were not described so far in the Czech Republic. METHODS AND RESULTS: The authors examined IgG antibodies (ELISA) against Helicobacter pylori in 411 subjects incl. 102 children with non-ulcerative or ulcerative dyspepsia and in 115 children of similar age but without gastrointestinal complaints and in 194 family members of children with dyspepsia. The specificity of the ELISA test is 97.1% and the sensitivity 92.1% (discrimination analysis). Children with dyspepsia had more frequently a positive finding of IgG antibodies against H. pylori (p = 0.0001), as compared with the control group of children of the same age but without dyspepsia. A statistically significant source of infection with H. pylori for dyspeptic children was the mother (p = 0.0006) and both parents (p = 0.0069), who were also a source of infection for other family members (p = 0.0003 and 0.0084). The most frequent positive IgG finding was in fathers (76.7%), but the father was not a statistically significant source of infection for the examined children (p = 0.6120) nor for other members of the family (p = 0.2330). IgG positive mothers had 85.2% positive husbands, IgG negative mothers had 68.4% positive husbands (p = 0.1601). The frequency of IgG antibodies in the mother and father (in couples) was thus statistically independent. However, there was a significant correlation (p = 0.01) of (P/N) IgG antibodies in couples--i.e. father and mother of the examined children. Siblings of IgG positive children had a significantly higher mean value of antibodies (p = 0.05) and were older. CONCLUSIONS: The presented facts provide evidence that in the investigated group the Helicobacter pylori infection is more frequent in children with the upper dyspeptic syndrome than in children of similar age without gastrointestinal complaints. In families of children with dyspepsia the infection aggregates and is transmitted. The mother and both parents are involved in the transmission to children. Nevertheless it is probable that there is also mutual transmission in children and from adults to children outside the family and in another environment.

Distribution of silver stained proteins (SSPs) in ring-shaped nucleoli of leukemic lymphocytes.

The distribution of silver stained proteins (SSPs) was investigated in ring shaped nucleoli of lymphocytes of the peripheral blood of patients suffering from chronic lymphocytic leukemia (B type). As expected, SSPs were present in fibrillar centers and adjacent nucleolar regions which appeared as bud and protrusion-like structures. The inhibition of the nucleolar RNA transcription by cultivation of leukemic lymphocytes at low temperature (4 degrees C) resulted in a significant reduction of these structures at fibrillar centers of ring-shaped nucleoli and decreased the number of cells which contained a large number of such structures at fibrillar centers of their ring shaped nucleoli. There was no substantial difference in the visualization and distribution of SSPs after fixation with phosphate-buffered glutaraldehyde or formaldehyde in distilled water and postfixation in ethanol or methanol-glacial acetic acid mixture.

Isolation and preliminary evaluation of a low-molecular-mass antigen preparation for improved detection of Helicobacter pylori immunoglobulin G antibodies.

Previously, immunoglobulin G (IgG) antibodies to five antigens with a relative molecular mass of between 15 and 30 kDa from Helicobacter pylori were found to be significantly more frequent in H. pylori-infected patients than in noninfected patients. In this study, these specific low-molecular-mass (LMW) antigens were separated by ultrafiltration of whole-cell sonicates. The LMW antigen preparation was evaluated by enzyme-linked immunosorbent assay with serum samples from 76 children with abdominal symptoms and 151 adults with dyspeptic symptoms. H. pylori was cultured or seen in 40 (53%) children and 83 (55%) adults. Increased antibody levels to H. pylori were found in serum from 35 (46%) children and 88 (58%) adults. Values for sensitivity, specificity, and predictive value of positive and negative results of the test were higher with LMW antigens than with the heat-stable antigen previously described. The low specificity and predictive value of a positive result were due to seropositive results for 21 persons with a negative culture for H. pylori and negative microscopy results for Helicobacter-like organisms in biopsies from gastric mucosa. Histologically, chronic gastritis was demonstrated in 43% of these persons, and 19% had peptic ulcer, indicating that they have or have had H. pylori infection. Specific antibodies to H. pylori were confirmed in all 21 patients by the Western immunoblot technique. Use of the LMW antigen improved the IgG antibody detection in patients with H. pylori infection, even though the results reflect the difficulties in establishing a true gold standard for diagnosis of H. pylori infection.

Glycosaminoglycan polysulfuric acid (GAGPS) in osteoarthritis of the knee.

We studied the efficacy and tolerability of glycosaminoglycan polysulfuric acid (GAGPS) in 80 patients with osteoarthritis (OA) of the knee. Patients received two series of five intra-articular injections, at 1-week intervals, of 25 mg (0.5 ml) GAGPS into the knee in a double-blind, parallel, randomized, placebo-controlled trial. There was an immediate decrease in pain after the injections of 43% with GAGPS and 33% with placebo (P = 0.047) (Jezek pain index). Pain relief of GAGPS vs placebo was not different at other intervals (10, 14, 22, 26 weeks after start of treatment). At 6 weeks the Lequesne index decreased 20% after GAGPS and 9% after placebo (P = 0.17). At 10 weeks the Lequesne index decreased 24% after GAGPS and 13% after placebo (P = 0.20). The decrease in Lequesne index at 14 weeks was 31% after GAGPS and 15% after placebo (P = 0.06). The other measured parameters tended to be more favorably influenced by GAGPS than placebo. GAGPS was well tolerated, with associated mild adverse reactions in 8% of cases. GAGPS may have a role as a symptomatic slow acting drug for OA. Further study appears appropriate.

Correlation between knee roentgenogram changes and clinical symptoms in osteoarthritis.

The main purpose of this study was to analyze the relationship between clinical symptoms and roentgenographic changes in patients with incipient osteoarthritis of the knees (OA). One hundred and sixty-two patients (126 women, 36 men, mean age 59 years) with established osteoarthritis of the knee were examined. Patients performed subjective self-assessment using Lequesne's questionnaire (index of severity for OA of the knee) and had frontal and lateral roentgenograms of both knees. Validity of the indices as reflections of individual symptoms was documented. A statistically significant correlation was found between clinical and patient indices (r = 0.327, p < 0.0001) as well as a very weak correlation between the clinical and roentgenogram indices (r = 0.196, p = 0.286). However, there was no correlation between the patient index and roentgenogram index (r = 0.065, p = 0.47). For the assessment of severity and progression of the disease, items reflecting acute irritation (swelling, effusion, increased joint temperature, pain during passive motion) were more reliable than items reflecting chronicity (muscular atrophy, decreased range of motion, crepitus). This study confirmed that Lequesne's questionnaire is a suitable tool for assessing subjective symptoms as well as for generating a condensed measure of disease severity. In addition, sclerosis, narrowing and osteophytes were proved to be important features characterizing the OA process.

[Serologic detection of antibodies to Helicobacter pylori in children and adolescents using ELISA]. / Sérologický prukaz protilátek proti Helicobacter pylori u deti a dospívajících metodou ELISA.

Serum antibodies anti-Helicobacter pylori (H.p.) which are specific against antigenic determinants of the bacterial body, were determined in 183 non-ulcer dyspeptic children and adolescents, aged 7- to 20 years. 108 patients (59%) of the same group underwent fibroendoscopy for direct detection of H.p. in bioptic specimens of the stomach and duodenum microbiological methods. Discrimination analysis revealed a overall positive correlation between the results of the two methods: P < 0.01. For detection of specific IgG antibodies against H.p., evaluation by the P/N ratio was used, i.e. the absorbance of the examined serum (P) was compared with the mean absorbance of three negative serum controls (N). The specificity of the ELISA test in this examination was 96.1%, the sensitivity of 90.3%. The mentioned ELISA method for detection of specific IgG antibodies against H.p. is comparable with direct microbiological prove of H.p. presence in mucus specimens in our group examined. It is a very important non-invasive method useful for primary detection of H.p. infection.

Ultrastructure of the cumulus oophorus in the mouse Graafian follicle during the preovulation period.

The author studied the ultrastructure of the cumulus oophorus in three groups of Graafian follicles divided according to the degree of maturity of the oocyte: follicles containing an oocyte with a germinal vesicle (group I), follicles containing a metaphase I oocyte (group II), and follicles containing a metaphase II oocyte (group III). Loosening of contacts between the corona radiata cells and the oocyte, as well as intercellular contacts of the cumulus oophorus cells, was observed after the resumption of meiosis and the deposition of finely granular material in the intercellular spaces (expansion and mucification). The cytoplasm of all the cells contained organelles typical of protein synthesis (granular endoplasmic reticulum, a large number of free ribosomes, a Golgi apparatus). There was practically no smooth endoplasmic reticulum. The inner mitochondrial membranes had processes in the form of cristae. Lipid droplets were present in small numbers. The morphology and number of the cytoplasmic organelles remained similar during the whole of the given period, with the exception of the lipid droplets, the number of which was slightly higher in group III. No morphological signs indicative of steroid hormone synthesis were found. During the whole of the preovulation period, the character of cellular proteosynthetic activity remained the same.

[Determination of antibodies to Campylobacter pylori in children and adolescents with gastroduodenal disorders]. / Stanovení protilátek proti Campylobacter pylori u detí a dorostu s gastroduodenálními obtízemi.

Serological examination of antibodies against Campylobacter pylori was made in 195 boys and girls aged 9-20 years. In all on account of gastrointestinal complaints endoscopy was indicated. The results of microbiological and histological finding), in particular in the antrum and corpus of the stomach). Positive titres antrum and body of the stomach were compared with results of the complement fixation test and the ELISA reaction. Antibodies against C. pylori were detected in 64 of the examined subjects. The titres assessed by the complement fixation and ELISA test were in good agreement. Patients with positive sera had significantly more often a positive endoscopic finding (bacteria, chronic gastritis) and bacteriological finding), in particular in the antrum and corpus of the stomach). Positive titres of the complement fixation reaction persisted frequently for 15 months after the first examination. From the results ensues that serological examination of antibodies against C. pylori can be considered a sparing examination method which in patients with gastrointestinal complaints should precede endoscopic examination and moreover it can be used to follow up the course of the disease and success of therapy.

[Can epigastric pain and non-ulcerative dyspepsia in children and adolescents be Campylobacter pylori infection?]. / Mohou byt bolesti v epigastriu a neulcerózní dyspepsie u detí a dospívajících infekcním onemocnením Campylobacter pylori?

The authors examined 263 children and adolescents aged 5-20 years who suffered from so-called non-ulcerative dyspepsia or epigastric pain, because of the suspected presence of Campylobacter pylori. In 31.9% of the examined subjects direct microbiological methods revealed its presence in the gastroduodenal mucosa. The confidence limit of positive results of Campylobacter pylori in the population is between 27.58% and 38.84%, the probability being 95%. The authors proved a mutual correlation between endoscopic, histological, serological findings and detection of Campylobacter pylori by microbiological methods. The statistical significance of the correlation of campylobacter pylori and chronic gastritis B (mostly inactive) provides further support for the hypothesis of the aetiological role of Campylobacter pylori in the development of chronic gastritis B in children and adolescents. From the investigation it does not ensue, however, that colonization with Campylobacter pylori is associated with certain clinical symptoms in all instances. However, in the differential diagnosis of so-called non-ulcerative dyspepsia and epigastric pain in children and adolescents we must include infection with Campylobacter pylori among their possible causes.

[Changes in NK cell activity in patients with acute leukemia]. / Zmeny v aktivite NK bunek u nemocných s akutní leukémií.

Examination of the activity of NK-cells in peripheral mononuclear cells (PMNB) of patients with acute leukaemia (AL) was implemented before the onset of treatment and in the course of the disease following cytostatic treatment. The examination was made by the isotope method based on the release of 51Cr after 4-hour cultivation of effector cells with target cells of line K 562. The mean percentage of NK cellular activity in patients with AL was 9.9 and 5.9, as compared with normal values of 56 and 41 with ratios of effector and target cells of 40:1 and 20:1. The course of the disease after cytostatic treatment revealed that patients where complete remission occurred had before onset of treatment on average a higher NK-cell activity than those who responded poorly to treatment. In the majority of patients who were in complete remission the activity of NK-cells increased markedly.