[The new blood test bio-marker SEPT9 and colorectal carcinoma screening]. / Nový krevní test bio-markeru SEPT9 a screening kolorektálního karcinomu.

Due to its high incidence and mortality rates, the colorectal carcinoma represents a crucial medical issue. However, when it is detected in early stage there is high rate of successful treatment. Thats why, early stage cancer screening programmes were introduced into the clinical practice. They focus on the finding of hidden bleeding, using various laboratory techniques, sigmoidoscopy, and, primarily, colonoscopy. However, screening programmes have not yet reached the effect required. New techniques are therefore being developed, such as the detection of blood bio-markers. This group includes also methylated SEPT9 (mSEPT9) detection in blood. We applied this test on 57 patients; we divided the group into two parts. There were 33 asymptomatic individuals in the first group. In this group, we were got only one positive mSEPT9 result. The consequent colonoscopies were negative. The other group had 24 proven carcinomas. Of them, two had negative mSEPT9 results. The remaining in all 22 patients was tested mSEPT9 positive. After its efficiency is tested by further studies, this test may be used especially for patients with low compliance, as it only requires routine blood drawing.

Probiotic treatment with Probioflora in patients with predicted severe acute pancreatitis without organ failure.

BACKGROUND: We previously demonstrated that probiotic prophylaxis, in patients with predicted severe pancreatitis, did not prevent infectious complications but unexpectedly increased the risk of bowel ischemia and mortality. The suggestion that these negative findings are only observed in the presence of organ failure at the start of probiotic treatment has not been confirmed. METHODS: In a retrospective analysis, all patients with predicted severe acute pancreatitis without initial organ failure admitted to a medium care facility of a teaching hospital in Prague from January 2003 to December 2010 were included. All patients routinely received probiotic treatment with Probioflora. Total parenteral nutrition (TPN) was routinely started and shifted toward total enteral nutrition. Infectious complications, mortality and the incidence of bowel ischemia were recorded. RESULTS: 99 consecutive patients, mean age 56 years, were included. Infectious complications occurred in 42 patients (42%), consisting of bacteremia (n = 40), pneumonia (n = 11) and infected necrosis (n = 11). Bowel ischemia was detected in two patients (2%). Overall mortality was 8%. CONCLUSION: In this retrospective study no apparent positive or negative impact of probiotic treatment with Probioflora was demonstrated when administered to patients with predicted severe acute pancreatitis without initial organ failure.

Calreticulin is a B cell molecular target in some gastrointestinal malignancies.

Calreticulin, upon translocation to the cell surface, plays a critical role in the recognition of tumour cells and in experimentally induced cellular anti-tumour immunity. However, less is known about anti-calreticulin antibodies and their role in malignancies. Using enzyme-linked immunosorbent assay (ELISA), we found immunoglobulin (Ig)A and/or IgG anti-calreticulin antibodies in sera of approximately 63% of patients with hepatocellular carcinoma (HCC), 57% of patients with colorectal adenocarcinoma (CRA) and 47% of patients with pancreatic adenocarcinoma (PACA), while healthy controls, patients with viral hepatitis C and with chronic pancreatitis reached only 2%, 20% and 31% seropositivity, respectively. We found significantly elevated mean levels of IgA anti-calreticulin antibodies (P < 0.001) in patients with HCC (78.7 +/- 52.3 AU, mean +/- standard deviation), PACA (66.5 +/- 30.9 AU) and CRA (61.8 +/- 25.8 AU) when compared to healthy controls (41.4 +/- 19.2 AU). Significantly elevated mean levels of IgG anti-calreticulin antibodies (P < 0.001) were detected in patients with HCC (121.9 +/- 94.2 AU), gall bladder adenocarcinoma (118.4 +/- 80.0 AU) and PACA (88.7 +/- 55.6 AU) when compared to healthy controls (56.7 +/- 22.9 AU). Pepscan analysis revealed a large number of antigenic epitopes of calreticulin recognized by both IgA and IgG antibodies of patients with HCC and PACA, indicating robust systemic immune response. Moreover, significantly elevated levels of antibodies against peptide KGEWKPRQIDNP (P < 0.001) in these patients, tested by ELISA, confirmed the distinct character of antibody reactivity against calreticulin. The high occurrence and specificity of serum anti-calreticulin autoantibodies in the majority of patients with some gastrointestinal malignancies provide the evidence for their possible clinical relevance.

Histologic findings after sodium phosphate bowel preparation for colonoscopy. Diagnostic pitfalls of colonoscopic biopsies.

Oral sodium phosphate (NaP) has been increasingly used for bowel preparation before the colonoscopy because it shows good patients tolerance and effective bowel cleansing ability. However, new studies describe that NaP can induce colonic mucosal damage. For better characterization of these changes, we examined histologically segmental colonic biopsies from 42 patients receiving NaP bowel solution before the colonoscopy. The series includes 25 male and 17 female patients in age from 19 to 81 years (average age 46.7 ys). Clinical symptoms in 37 patients included diarrhea, constipation, bleeding and abdominal cramps. The most frequent reason for colonoscopy was suspicion of microscopic colitis. Five patients underwent endoscopy to rule out the presence of neoplasia. None of the patients took drugs before the colonoscopy. Histologically, all specimens showed mild focal edema, hyperemia and hemorrhages. In addition to edema and hemorrhage, in 26 patients (61.9%), patchy mononuclear infiltration in the upper part of lamina propria and increased epithelial cell proliferation of individual crypts were seen. Mucosal structure was normal, with partial sloughing of normal or flattened surface epithelium. In 5 patients (11.9%), some biopsy samples contained scattered neutrophilic leucocytes in the lamina propria/superficial epithelium, isolated basal cryptitis, increased proliferation and apoptosis of the crypt epithelium. In two patients with focal cryptitis (4.8%), small erosions were found. Mild basal cryptitis, increased proliferation and striking apoptosis were present in two inflammatory pseudopolyps (in two patients). In 4 patients, solitary tubular adenomas with low-grade dysplasia without any reactive changes were found. In addition, 300 hyperplastic polyps removed endoscopically after the NaP application, were examined. Two polyps (0.75%) showed cryptitis and isolated multinucleated epithelial cells in the superficial part of the crypts. Our results are similar to those previously described in other studies of colonic changes after the NaP application. It reflects probably a similarity in composition of used NaP solutions.

Anti-calreticulin immunoglobulin A (IgA) antibodies in refractory coeliac disease.

Refractory coeliac disease (RCD) is a very rare and dangerous form of CD, in which gluten-free diet loses its therapeutic effect and the damage of intestinal mucosa persists. Because of the adherence to the diet, serological markers of CD [immunoglobulin A (IgA) antibodies against gliadin, tissue transglutaminase (tTG) and endomysium] are often missing in RCD patients. We found substantially elevated levels of IgA anti-calreticulin (CRT) antibodies in the sera of almost all RCD patients tested. These sera were negative for IgA antibodies to gliadin and tTG and only some of them showed IgA antibodies to enterocytes. Analysis of patients' IgA reactivity to CRT fragments (quarters and halves) by Western blotting revealed differences in the specificity of IgA antibodies between RCD and CD patients. We therefore used the Pepscan technique with synthetic overlapping decapeptides of CRT to characterize antigenic epitopes recognized by serum IgA antibodies of RCD patients. Employing this method we demonstrated several dominant antigenic epitopes recognized by IgA antibodies of RCD patients on the CRT molecule. Epitope GVTKAAEKQMKD was recognized predominantly by serum IgA of RCD patients. Our results suggest that testing for serum IgA antibodies against CRT and its selected peptide could be a very useful tool in RCD differential diagnosis.

Histopathologic changes in gastroesophageal reflux disease. A study of 126 bioptic and autoptic cases.

The histologic diagnosis of reflux esophagitis is still complicated by the lack of a consensus opinion on what is the normal mucosa in the area of the gastroesophageal junction (GEJ). Most authors consider GEJ as the junction between the squamous and the cardiac epithelium. The cardiac mucosa is composed of mucinous or mixed mucinous-oxyntic glands. These glands are in fact indistinguishable from metaplastic mucosa that arises in the distal esophagus in consequence of gastroesophageal reflux (GER). The cardiac mucosa shows invariably chronic inflammatory changes referred to as "carditis". The cause of "carditis" is GER and/or Helicobacter pylori (HP) infection. In our series of 120 endoscopic biopsies of the GEJ and distal esophagus the cardia type mucosa (CM) was always present. In 15 cases, it was accompanied by oxyntocardiac mucosa. Both mucosa types showed chronic inflammation that is after exclusion of HP infection regarded as a strong diagnostic sign of the gastroesophageal reflux disease (GERD). In two cases with clinical symptoms of GERD, a few HP were found on the CM. Therefore we diagnosed them as GERD with secondary HP infection. In 17 cases, CM displayed intestinal metaplasia (IM) predominantly of incomplete type and no dysplasia. This IM expressed MUC6 in the glandular zone of the mucosa like it did in the neighboring glands, whereas in the surface and foveolar epithelium the MUC6 was negative or only slightly and focally positive. On the other hand, IM in the surface and foveolar epithelium was reactive for MUC5AC. The positivity and distribution of CK7 and CK20 was very similar in the Barrett's mucosa, cardiac mucosa and antral mucosa. In one specimen of esophagus resected for adenocarcinoma, CM with incomplete IM was found in the vicinity of the tumor. Squamous metaplastic epithelium was often seen near the orifices of submucosal esophageal glands in these areas, indicating the metaplastic nature of the glandular mucosa in the distal esophagus. In the GEJ of 5 autopsy cases of children with spastic quadriplegia (age range 7-10 years) CM in a short segment (0.5-3 mm in length), probably of metaplastic origin was identified, showing chronic inactive inflammation.

[Endoscopic cytoscopy in examination of the digestive tract]. / Význam endoskopické cytoskopie u vysetrení trávicího traktu.

A number of new endoscopic methods have been developed recently, which aim to allow the most accurate possible viewing of the mucosa of the digestive tract. These procedures include endoscopic cytoscopy, which together with confocal endoscopy is a technique of so-called endoscopic microscopy. By means of enodcytoscopy it is possible to view "in vivo" the morphological details of the surface of the mucosa of the digestive tract. The mucosa must, however, be thoroughly cleaned and stained with methylene blue. The paper presents our own initial experience of this method, which may lead to faster and more accurate diagnosis of pre-tumorous or tumourous inflammation processes in the mucosa of the digestive tract.

[Pseudomembranous colitis]. / Pseudomembranózní kolitida.

For some thirty years it has been known that pathologic proliferation oftoxigenic Clostridium difficile may lead to inflammation of colonic mucosa which, in its fully developed form, manifests as pseudomembranous colitis. CDAD (Clostridium difficile - Associated Disease or Diarrhoea) is the term which is generally and quite aptly used for the disease in literature on the subject. In most cases, the disease develops after the administration of broad-spectrum antibiotics, yet there are also other important pathogenetic factors involved which should not be ignored. The disease has grown in significance over recent years due to the occurrence of serious epidemics in a number of advanced countries provoked by a highly virulent strain characterised as ribotype 027, toxinotype III, pulsovar NAP1. CDAD is a potentially fatal disease, yet it can be effectively cured if diagnosed on time. Stool toxin testing plays a crucial role in the diagnostics of the disease, and the use of endoscopy has been on the rise. Administration of targeted antibiotics is of critical importance for successful therapy, metronidazol or orally administered vankomycine being considered the most reliable, and there are also other drugs with promising effects. However, the rate of recurrence of the disease is as high as 25%. In view of the realistic threat of propagation of the highly virulent strain in the territory of the Czech Republic, preparations have started for the setting up of a national reference laboratory for Clostridium difficile typification.

[Complications of laparoscopic procedures of inguinal hernias, diagnosed on colonoscopy]. / Komplikace laparoskopické operace tríselné kýly diagnostikované kolonoskopicky.

AIM OF THE STUDY: The aim was to assess significance of colonoscopy in detection of inguinal hernias - dislocation of the mesh into the lumen of the large intestine, following inguinal hernia laparoscopic procedures (TAPP). MATERIAL, METHODOLOGY AND RESULTS: From 01-01-2004 to 31-12-2006, the authors performed 3 colonoscopies in 3 subjects after TAPP inguinal hernia laparoscopic procedures. The subjects included 2 males and 1 female, 4 to 9 months after the laparoscopic procedure, colonoscopy was performed, which was indicated for the lower abdominal pain and enterorrhagia. However, no signs of a relapsing hernia were recorded. Endoscopy detected irritation and bleeding of the sigmoid, and a mesh, which passed through the intestinal wall was detec ted in the sigmoid lumen. The subjects were then reoperated and the mesh was removed. In one subject, resection of the sigmoid had to be performed as well. The subject recovered. CONCLUSION: The postoperative complications of the TAPP procedure included dislocation of the mesh into the intestinal lumen. Detection of the mesh dislocation into the lumen of the large intestine facilitates indication for surgical management.

[Imported cryptococcal infection mimicking primary sclerosing cholangitis]. / Importovaná kryptokoková infekce imitující primární sklerozující cholangoitidu.

Expanding travel industry may increase the incidence of some rare infections diseases. One of them is cryptococcosis developing especially in immunosuppressed individuals. However, massive cryptococcal infection may lead to a severe clinical course even in people without impaired immunity. The case report describes the disease in a non-immunosuppressed 26-year-old man who returned to the Czech Republic after a 2.5-month stay in Egypt. He was hospitalized for increasing fever, weight loss and changes in behaviour. Thorough examination including thoracoscopy and histological analysis of the mediastinal lymph nodes revealed cryptococcal disease. Various organs, mainly the small bile ducts, were affected by the dissemination mimicking primary sclerosing cholangitis. The discussion is concerned with the properties of the Cryptococcus, variety of clinical manifestations, differential diagnosis and treatment.

Autoimmune gastritis. A clinicopathologic study of 25 cases.

The histopathological diagnosis of autoimmune gastritis (AG) in its early stages can be a diagnostic challenge. Even some advanced cases with complete atrophy of the corpus mucosa may be difficult to recognize. To establish the diagnosis of autoimmune gastritis, several histological features should be assessed and combined with immunostains for enterochromaffin cell-like (ECL) cells and G-cells. The main histological criteria include a mononuclear infiltrate within the lamina propria, foci of destruction of oxyntic glands, intestinal metaplasia (IM), pyloric metaplasia, and parietal cell pseudohypertrophy. These criteria were evaluated in our series of 25 patients with achlorhydria and/or megaloblastic anemia. Some of our patients presented with nonspecific gastrointestinal symptoms. The age ranged between 46 and 79 years; one male patient was only 31 years old. Histologically, the corpus mucosa displayed in all cases chronic inflammation with focal complete IM and advanced pyloric metaplasia. In 4 patients, oxyntic glands were destructed in some sites. There was a pancreatic metaplasia of acinar type in 2 patients and a minimal focal pseudohypertrophy of parietal cells in the 31-year-old man. A tubular adenoma with a low-grade dysplasia was found in one female patient. Immunohistochemically, chromogranin-A highlighted linear or nodular hyperplasia of ECL cells in 19 patients, and adenomatoid ECL hyperplasia in one case (80%). In the remaining cases hyperplasia of ECL cells could not be recognized from their normal count. In 13 cases (52%) a few ECL cells were seen also in IM. Regarding associated pathology, in one woman with nodular ECL cell hyperplasia, a gastric carcinoid was removed endoscopically. The reaction with gastrin antibody revealed in 11 cases (44%) a small number of G cells in IM in the corpus mucosa. In 18 patients, antral mucosa was examined as well. In 8 patients, the mucosa was normal; in 10 cases, a mild chronic inactive gastritis was diagnosed, and in 15 patients G-cell hyperplasia was found. In accordance with other studies, we show that the diagnosis of AG may be established microscopically in endoscopic specimens of the gastric body mucosa when histologic features and immunohistochemical detection of ECL and G cell hyperplasia are combined.

Phonon density of states of single-wall carbon nanotubes

The vibrational density of states of single-wall carbon nanotubes (SWNT) was obtained from inelastic neutron scattering data from 0 to 225 meV. The spectrum is similar to that of graphite above 40 meV, while intratube features are clearly observed at 22 and 36 meV. An unusual energy dependence below 10 meV is assigned to contributions from intertube modes in the 2D triangular lattice of SWNT bundles, and from intertube coupling to intratube excitations. Good agreement between experiment and a calculated density of states for the SWNT lattice is found over the entire energy range.

[Ulcerative colitis--contemporary morphological criteria]. / Ulcerózní kolitida--soucasná morfologická kritéria.

Regular bioptical examinations of patients with ulcerative colitis (UC) performed in recent years show that the inflammatory changes of the mucosa of the large intestine are not necessarily diffuse, and that their extent may vary in the course of the disease. To establish the diagnosis of UC and to assess the treatment efficacy it is important to examine histologically multiple mucosal specimens from different levels of the large intestine. In our series of 27 patients with ulcerative colitis (18 men and 9 women at the age of 17 to 76 years), active or active and inactive pancolitis was diagnosed in 25 cases (93%). In 11 of these, the whole of the large intestine was affected. Two patients showed diffuse pancolitis without caecal involvement, in 5 cases there was inactive inflammation in the rectum or in the sigmoid colon. Seven patients had active colitis of the rectum and sigmoid. In another 2 patients (7%), the inflammation was limited to several segments of the large intestine only (the descending colon, and the descending and transverse colon). On bioptical examination of 6 patients repeated after 2-29 months (mean 14 months), there were changes in the distribution and appearance of the inflammation. Thus our findings correspond with the results of previous studies: UC does not always affect the mucosa of the large intestine diffusely. Further, the extent and distribution of inflammatory changes vary in the course of the disease.

Occurrence of IgA and IgG autoantibodies to calreticulin in coeliac disease and various autoimmune diseases.

Calreticulin (CRT), a high-affintiy calcium binding protein and chaperone, was recently identified as one of the targets of autoantibodies in coeliac disease. We evaluated the level of IgA and IgG antibodies to CRT in sera from patients with coeliac disease and various autoimmune diseases. The level of antibodies to gliadin (shown previously to cross-react with CTR), isolated enterocytes and tissue transglutaminase were determined for comparison. The mean level of IgA antibodies to CRT was significantly higher (P< 0.001) in sera from coeliac patients with active disease (139.9+/-11.2 AU/+/-SE) than in healthy controls (20.9+/-1.7 AU). In sera of patients with systemic lupus erythematosus (SLE), insulin dependent diabetes mellitus (IDDM), multiple sclerosis (MS) and autoimmune thyroiditis (AT) or inflammatory bowel disease (IBD) the mean level (25.8+/-3.7 to 38.1+/-5.6 AU) did not exceed the cut-off value. A low level of these antibodies, however, was detected in some sera of patients with MS and IBD. The level of IgG anti-CRT antibodies was increased in coeliac patients (mean 125.4+/-8.0 AU, P< 0.001) when compared to that in healthy controls (33.9+/-2.3 AU). The IgG anti-CRT antibodies were also detected in about 30% of SLE patients sera (54.1+/-3.6 AU, P< 0.001), but the mean level reached only half that detected in coeliac patients.

Quantized phonon spectrum of single-wall carbon nanotubes

The electronic spectra of carbon nanotubes and other nanoscale systems are quantized because of their small radii. Similar quantization in the phonon spectra has been difficult to observe because of the far smaller energy scale. We probed this regime by measuring the temperature-dependent specific heat of purified single-wall nanotubes. The data show direct evidence of one-dimensional quantized phonon subbands. Above 4 kelvin, they are in excellent agreement with model calculations of individual nanotubes and differ markedly from the specific heat of two-dimensional graphene or three-dimensional graphite. Detailed modeling yields an energy of 4.3 millielectron volts for the lowest quantized phonon subband and a tube-tube (or "lattice") Debye energy of 1.1 millielectron volts, implying a small intertube coupling in bundles.