Gastrointestinal toxicity of systemic oncology immunotherapy.

BACKGROUND: Systemic anti-cancer immunotherapy provides a substantial progress in options of current oncology treatment. Yet, this therapeutic approach is potentially associated with a significant gastrointestinal toxicity. AIM: The purpose of this paper is to provide a comprehensive review on pathogenesis, clinical features, dia gnostics and therapy of these toxicities. Review of current knowledge: Check-point inhibitors brought a major progress in anti-cancer immunotherapy and improved significantly prognosis of several malignancies (e. g. metastatic malignant melanoma, non-small-cell lung cancer, gastric and colorectal cancers in high-risk population associated with presence of pathogenic mutations, renal cell carcinoma, squamous cell carcinoma of the head and neck and urothelial carcinoma). They include monoclonal antibodies targeting cytotoxic T lymphocyte-associated antigen 4 (CTLA4; e. g. ipilimumab, tremelimumab), programmed death-1 receptor (PD-1; e. g. pembrolizumab, nivolumab) and its ligand PD-L1 (e. gatezolizumab, avelumab). Chimeric antigen receptor (CAR) T-cell therapy is another new option for haematological malignancies and metastatic colorectal cancer. Major symptoms of gastrointestinal toxicity caused by systemic immunotherapy include diarrhoea (20-50%), entero-colitis (1-10%) and laboratory or clinical signs of hepatopathy (~10%). Anti-cancer immunotherapy can be also complicated by infections (Clostridium difficile, Mycoplasma and/ or cytomegalovirus). There is no data on other possible complications so far. However, it can be assumed that these will also include bile acid malabsorption as well as small intestinal bacterial overgrowth syndrome. Treatment of gastrointestinal complications of immunotherapy should be graded according to their severity. It includes symptomatic medications (e. g. loperamide), systemic glucocorticoids and anti-TNF monoclonal antibodies (alone or together with mycofenolate mofetil or tacrolimus in the most severe cases). CONCLUSIONS: Awareness of possible complications of systemic anti-cancer immunotherapy is crucial for patients safety. It is mandatory to consider immune-related adverse events, complicating infections, bile acids malabsorption and small intestinal bacterial overgrowth syndrome. Prompt proper dia gnostics and immediate vigorous therapy infl uence the outcome of patients signifi cantly. A strictly individualized approach is indispensable.

Anti-Outer membrane protein C antibodies in colorectal neoplasia.

Sporadic colorectal cancer (CRC) represents an enormous problem worldwide. Large intestinal microbiota play an important role in the colorectal carcinogenesis. The aim of the study was to investigate anti-Outer membrane protein C (anti-OmpC) antibodies, aimed at porin C, which is embedded in the outer membrane of gram-negative bacteria, in patients with colorectal adenoma (CRA), CRC and controls. The study included 22 patients with CRA (11 men, 11 women, aged 26-79, mean 65 ± 12), 11 patients with CRC (9 men, 2 women, aged 50-83, mean 66 ± 11) and 45 controls, blood donors (24 men, 21 women, aged 20-58, mean 38 ± 10). Serum anti-OmpC antibodies were investigated by means of ELISA. Values of 0-20 U/mL were considered to be negative; values >25 U/mL were assessed as positive. A total of 9/11 (82 %) patients with CRC had positive anti-OmpC antibodies. Anti-OmpC antibodies were negative or grey-zone in 37/45 (82 %) controls. Serum anti-OmpC were found to be significantly higher in patients with CRC (median 42.4, interquartile range (IQR) 22.2) compared to controls (median 18.3, IQR 12.4), p < 0.001. No statistically significant difference in anti-OmpC was found between controls (median 18.3, IQR 12.4) and CRA patients (median 17.7, IQR 16.5), p = 0.326. Anti-OmpC were significantly higher in patients with CRC (median 42.4, IQR 22.2) compared to patients with CRA (median 17.7, IQR 16.5), p = 0.011. Positivity of anti-OmpC antibodies was found in patients with CRC, which supports the contribution of gram-negative large intestinal microbiota to the pathogenesis of CRC.

Use of electrogastrography in preclinical studies of cholinergic and anticholinergic agents in experimental pigs.

Electrogastrography (EGG) is a non-invasive method for the assessment of gastric myoelectrical activity. Porcine EGG is comparable with human one. The purpose of this study was to evaluate the effect of atropine and neostigmine on the EGG in experimental pigs. Adult female pigs were administrated atropine (1.5 mg i.m., n=6) and neostigmine (0.5 mg i.m., n=6) after the baseline EGG, followed by a 90-min trial recording (MMS, Enschede, the Netherlands). Running spectral analysis was used for the evaluation. The results were expressed as dominant frequency of slow waves and EGG power (areas of amplitudes). Neostigmine increased continuously the dominant frequency and decreased significantly the EGG power. Atropine did not change the dominant frequency significantly. However, atropine increased significantly the EGG power (areas of amplitudes) from basal values to the maximum at the 10-20-min interval. After that period, the areas of amplitudes decreased significantly to the lowest values at the 60-90-min interval. In conclusion, cholinergic and anticholinergic agents affect differently EGG in experimental pigs.

Preparing Triple-Compound Heterozygous Control Material for Molecular Diagnostics of TPMT Allelic Variants.

The aim of the study is to present a novel approach for preparing triple-compound heterozygous reference material (TCH-RM) for thiopurine S-methyltransferase (TPMT) genotyping by using the gene synthesis technology. The polynucleotide chain we prepared consisted of three wild-type and three mutant segments corresponding to the TPMT 238G>C, 460G>A, and 719A>G polymorphic sites. TCH-RM characteristics were assessed via four methods: reverse hybridization, real-time PCR with hydrolysis probes, real-time PCR followed by subsequent melting temperature analysis, and DNA sequencing. Consequently, we investigated the TPMT genotype of 371 patients suffering from autoimmune diseases requiring immunosuppressive therapy with thiopurine drugs, mostly inflammatory bowel disease. All methods confirmed the triple heterozygous character and commutability of TCH-RM. In evaluating its stability we obtained very comparable data before and after six months of storage at -80 °C. The determined genotypes were as follows: 352 wild-type subjects (94.8%), 17 TPMT*3A heterozygotes (460G>A and 719A>G, 4.6%), one patient heterozygous for the TPMT*2 allele (238G>C, 0.3%), and one TPMT*3C heterozygote (719A>G, 0.3%). The frequencies of TPMT*1, *3A, *3C, and *2 in the patients were 97.5%, 2.3%, 0.1%, and 0.1 %, respectively. Assembling segments of synthetic DNA into long polynucleotide chains is a universal way of obtaining compound heterozygous material for performing any simultaneous analysis of polymorphic sites in the human genome. The batches are manufactured with a perfect concentration match of wildtype and mutant fragments, and can be made in large quantities for most diagnostic techniques.

Resistance to selected beta-lactam antibiotics.

Susceptibility in vitro and trends in resistance to antimicrobials were determined by a dilution micromethod in a group of Actinobacillus pleuropneumoniae, Pasteurella multocida, Mannheimia haemolytica and Escherichia coli isolates from clinical cases of cattle and swine diseases in the Czech Republic from 2007 to 2011. A high susceptibility of pig and cattle respiratory pathogens to antimicrobials was found, with the exception of the moderate prevalence of M. haemolytica resistance to ampicillin. In contrast to respiratory pathogens, low susceptibility of E. coli of pig and cattle isolates to ampicillin and amoxicillin/clavulanic acid was noted. Regarding resistance trends, an increase in levels of resistance among E. coli isolates to ampicillin and amoxicillin/clavulanic acid was identified, but the resistance of respiratory isolates was low, with the exception of M. haemolytica. For the period of 2007-2011, there was a significant and almost continuous increase in sales (compared with population correction unit) of ceftiofur, cefquinome and other beta lactams for pigs. Consumption peaked in 2010. In the case of amoxicillin in combination with clavulanic acid, data showed a significant decrease in sales from 2007 to 2008, followed by a period of fluctuation. In cattle, within the groups of 3rd and 4th generation cephalosporins and for the whole group of other betalactams for the period of 2007-2011, there was a significant and almost continuous increase in sales (compared with population correction unit). Consumption peaked in 2010. In the case of ceftiofur, there was a huge increase noted from 2010. In the case of amoxicillin in combination with betalactamase inhibitor (clavulanic acid) data shows a significant decrease from 2007 to 2008, followed by a period of fluctuation in sales.

Comparison of male and female rats in avoidance of a moving object: more thigmotaxis, hypolocomotion and fear-like reactions in females.

Although male rats generally outperform females in many spatial tasks, sometimes gender differences are not present. This preliminary study examined gender effects in the Enemy avoidance task, in which a rat on a stable circular arena avoids approaching a small mobile robot while collecting randomly dispersed small pellets. Whenever distance between robot and the rat dropped below 25 cm, animal was punished by a mild footshock. Female rats showed thigmotaxis, hypolocomotion and avoidance of robot in the habituation phase, when approaches were not punished. No statistically significant differences in avoidance learning under reinforcement training sessions were observed; but females still spent significantly more time at periphery of the arena and foraged less than males. We conclude that females were able to perform at the same level as males under reinforcement despite different behavioral strategy. The thigmotaxic behavior appears to function as innate escape strategy in female rats triggered by the stressing effect of the moving robot rather then the presence of shocks.

The importance of upper gastrointestinal lesions detected with capsule endoscopy in patients with obscure digestive bleeding.

STUDY AIMS: Small bowel capsule endoscopy (SBCE) is the first line procedure for detecting small bowel lesions in patients with an obscure gastrointestinal bleeding (OGIB). Missed upper gastrointestinal (UGI) lesions at the initial endoscopy may account for the so-called OGIB. This retrospective study was designed to assess the role of SBCE in detecting missed UGI lesions. METHODS: All consecutive SBCE that were performed in the last year for patients with OGIB were included in our study. We evaluated the visibility of the gastric mucosa, the anatomic landmarks, the presence of UGI lesions as well as their clinical importance. The SBCE findings were compared with the reports of previous UGI endoscopies. RESULTS: 118 patients (45 males, 73 females, mean age 61 +/- 19 years) were included in the analysis. The indication for SBCE was obscure overt and occult OGIB in 60 and in 58 patients, respectively. SBCE identified lesions in the small bowel in 42% of the patients. An excellent visibility of gastric mucosa was observed in 83/118 cases (703%). SBCE identified gastric lesions with potential clinical significance (high bleed potential) in 25/118 (21.2%) patients. In 12/118 (10.2%) patients the UGI lesions detected by SBCE were considered as the only potential source of bleeding. CONCLUSIONS: In patients with OGIB, SBCE detected not only small bowel lesions but also significant UGI lesions that were missed or underestimated at the initial endoscopy in 21% of cases. It is therefore necessary to carefully read the gastric images when performing an SBCE.

[Peptic ulcer disease in patients with diabetes mellitus]. / Gastroduodenální vredová choroba u diabetika.

Peptic ulcer disease and diabetes mellitus are the serious chronic diseases with many medicals and socioeconomic consequences. Although the changes leading to the mucosal vulnerability and prolonged healing of ulcers were many times demonstrated in experiment, the clinical data does not confirmed explicit association of those diseases. The patients with diabetes mellitus are threatened by complications of peptic ulcer disease (bleeding, perforation) above all. The Helicobacter pylori prevalence is in diabetics higher, the results of eradication of Helicobacter pylori are unsatisfactory.

[Motility disorders and gastric emptying in diabetes mellitus. Current diagnostics and treatment]. / Poruchy motility a evakuace zaludku u diabetika. Soucasné moznosti diagnostiky a lécby.

Autonomic neuropathy of the gastrointestinal tract is a common complication of diabetes mellitus. However, it is seldom recognised properly as it is rarely considered. In some patients, it might be asymptomatic or hardly compensatable diabetes can be the only one sign. There are non-invasive diagnostic methods to assess gastric emptying rate (13C-octanoic acid breath test) or myoelectric activity of the stomach (electrogastrography). The principle treatment comprises optimal control of diabetes and use of prokinetics.

Biodegradable stents for the treatment of benign stenoses of the small and large intestines.

Biodegradable stents, which are made of various synthetic polymers, such as polylactide or polyglycolide, or co-polymers, such as polydioxanone, can be used for the treatment of benign refractory stenoses of the gastrointestinal tract. Here we report 11 patients (median age 41) with stenosing Crohn's disease of the small and/or large intestine. Endoscopic insertion of a biodegradable stent was successful at the first attempt in all patients except one. Subsequent follow-up was for a mean of 16 months, median 17 months, range 12-29 months. Early stent migration (between 2 days and 8 weeks) was seen in three patients. Mucosal overgrowth (epithelial hyperplasia) was not observed in any of the patients during the follow-up period. The high rate of early stent migration might be solved by appropriate tailoring and further improvements in the design of the biodegradable stents. Proof of long-term efficacy and safety requires further studies.

Morphometric analysis of the porcine gastrointestinal tract in a 10-day high-dose indomethacin administration with or without probiotic bacteria Escherichia coli Nissle 1917.

BACKGROUND: Nonsteroidal anti-inflammatory drugs may cause severe injury to all parts of the gastrointestinal tract. It has been hypothesised that probiotic bacteria might reduce this adverse effect. The aim of this study was to perform a morphometric evaluation of the gastrointestinal tract in experimental pigs treated using a 10-day high-dose of indomethacin with or without Escherichia coli Nissle 1917 (EcN). METHODS: Twenty-four healthy mature pigs were included: Group A (controls; 6 animals), Group B (EcN; n = 6), Group C (indomethacin; n = 6) and Group D (EcN & indomethacin; n = 6). EcN (3.5 × 10(10) live bacteria/day for 14 days) and/or indomethacin (15 mg/kg/day for 10 days) were administered. Specimens of the stomach, small and large bowel were routinely processed for microscopic examination. The height of glandular mucosa, height and width of interfoveolar spaces and villi and basement size of epithelial cells were evaluated. RESULTS: Different effects of indomethacin and EcN on particular parts of the gastrointestinal tract were shown. The indomethacin and probiotics group demonstrated a significantly lower height of cryptal mucosa and colonocytes and widening of the basement size of colonocytes compared to controls (p = 0.004; p < 0.001; p = 0.025). The height of cryptal mucosa was significantly higher in the EcN group compared to controls (p = 0.001). CONCLUSIONS: Indomethacin alone induced marked adaptation of the gastric mucosa. EcN alone provided a significant favourable trophic effect on the colonic mucosa. However, indomethacin and probiotics administered together comprise the worst impact on all porcine stomach, small and large bowel.

[Epidemiology of Helicobacter pylori infection]. / Epidemiologie Helicobacter pylori.

Helicobacter pylori (Hp) is the most common chronic bacterial infection in humans worldwide. The prevalence of Hp infection is high in developing countries (80-90%) and lower in developed countries (10-30%). In vast majority of infected individuals, the infection is acquired early in life. The risk of Hp infection is related to low socio-economic status and living conditions at early childhood (density of housing, overcrowding, number of siblings, sharing a bed, and lack of running water, low education of parents). Smoking is a risk factor of Hp infection in adults. In developed countries including the Czech Republic, the overall prevalence of Hp infection has fallen dramatically over recent decades. This decrease can be explained mostly by the relatively favourable and improving socio-economic conditions. However, it is necessary to consider also the fundamental determinants of "modern times" that could cause gradual disappearing of Hp from the human "microbiome".

[Inadequate screening and low awareness about risk factors among first-degree relatives of colorectal cancer patients]. / Nedostatecný screening a malá informovanost o rizikových faktorech a príbuzných I. stupne nemocných s kolorektálním karcinomem.

INTRODUCTION: First-degree relatives ofcolorectal cancer patients have higher risk of disease, thus colonoscopic screening should be started at the age of 40 in this group. Goal of the study was to assess their awareness of the colorectal cancer risk factors and their compliance with colorectal cancer screening programme. METHODS: 62 patients after colorectal cancer resection and their 67 first-degree relatives (42% out of 160 addressed) answered structured questionnaire and counselling focused on colorectal cancer risk factors, and screening participation. RESULTS: Before diagnosis of colorectal cancer only 18% of colorectal cancer patients (11/62) were aware of its risk factors, after diagnosis their awareness increased to 65% (40/62, p < 0.001). Before questionnaire 46% of first-degree relatives (31/67) were aware of colorectal cancer risk factors, after counselling awareness increased to 66% (44/67, p = 0.024). 79% (53/67) offirst-degree relatives were aware of their increased familial risk. 28% (19/67) of first-degree relatives enrolled for colorectal cancer screening. Most frequent screening method was fecal occult blood test (FOBT) in 21% (14/67) of first-degree relatives, colonoscopy was performed only in 7% (8/67) of first-degree relatives. Average age of screening participation was 53 years. 22% (15/67) of first-degree relatives refused any screening in the future. CONCLUSIONS: We have found low awareness of colorectal cancer risk factors in colorectal cancer patients and their relatives before their diagnosis of disease respectively before their counselling. First-degree relatives are aware of their increased risk, however their screening participation is unsatisfactory. Screening is performed only in minority of first-degree relatives, often late and with inappropriate modality (FOBT). 22% of first-degree relatives refused any screening in spite of their increased colorectal cancer risk.

[Inflammatory bowel disease--familial and sporadic form]. / Idiopatické strevní zánety--familiární a sporadická forma.

THE AIM: The aim of this thesis was to elucidate more differences between a familial and sporadic inflammatory bowel disease by comparing certain clinical data. METHODS: We assessed 248 patients with inflammatory bowel disease (IBD) observed in 1994-2004 in the Academic Department of Gastroenterology at the Medical Faculty in Hradec Králové. To get information about the defined characters we obtained from the questionary and the hospital data. RESULTS: We did not identify any relationship between the onset of the disease and a certain age group, yet males seem to be more prone to familial Crohn's disease. The more frequent familial form of Crohn's disease was the fibro-stenotic one. There were no differences in the onset of the disease. We did not prove the differences in extraintestinal signs, alergy and comorbidities. We did not find any differences in therapy response in relation to the type of nutrition (enteral, parenteral) and the administration of immunosupresive drugs. The biological therapy in sporadic and familial Crohn's disease did not differ either. Surgical intervention was more frequent in Crohn's patients compared to the patients with ulcerative colitis; yet no difference was identified between familial and sporadic cases. Appendectomy carried out before the onset of the disease was later diagnosed as Crohn's disease in more instances than ulcerative colitis. CONCLUSION: We did not prove significant differences comparing certain clinical data in familial and sporadic form of inflammatory bowel disease, yet males seem to be more prone to familial Crohn's disease. Small bowel was involved more often in familial form of Crohn's disease than in sporadic form.

[Gastroesophageal reflux disease. Standards of the Czech Society of Gastroenterology--actualization 2009]. / Refluxní choroba jícnu. Standardy Ceské gastroenterologické spolecnosti--aktualizace 2009.

Presently, gastroesophageal reflux disease is defined as a disorder where reflux of the stomach content is bothersome and/or brings about complications. The state when macroscopically detectable erosions of mucosa are present is known as erosive reflux disease and the term non-erosive reflux disease is used for the condition with no macroscopic erosions. Reflux oesophagitis is a frequent sign of the disease. A condition, where reflux symptoms persist or new occur and oesophagitis healing fails to take place despite maximum treatment, is classified as refractory gastroesophageal reflux disease. The main symptoms of gastroesophageal reflux disease include heartburn and regurgitation. Gastroesophageal reflux disease may, less frequently, manifest itself with isolated non-oesophageal symptoms, e.g. recurring upper respiratory tract infections or bronchial asthma. Etiopathogenesis involves refluxate, motility disorders, altered anatomic proportions, protective mechanisms disorder and external factors. Diagnosis takes place on the basis of typical symptomatology and endoscopic examination. Complications include bleeding, ulceration, strictures and Barrett's oesophagus. Lifestyle and dietary measures are an important treatment approach as are pharmacological (antisecretion and prokinetic agents) as well as surgical management.

Confocal laser endomicroscopy in experimental pigs. Methods of ex vivo imaging.

BACKGROUND: Confocal laser scanning endomicroscopy (CLSE) enables online in vivo cellular surface and subsurface imaging of normal and pathological tissue at high resolution and magnification. The aim of this study was to work out a method of ex vivo in vitro CLSE in experimental pigs and to compare CLSE images with those of "classic" histology. MATERIAL AND METHODS: Five mature female pigs entered the study. CLSE on an ex vivo in vitro basis was started 10 minutes after pharmacological euthanasia and carried out for 30 minutes. Fluorescein was administrated i.v. as a fluorescence substance. RESULTS: CLSE was successful in all tissue samples of all animals (the oesophagus, stomach, small intestine, large bowel). We have succeeded to obtain high quality images within the first 30 minutes that means 40 minutes after the euthanasia of experimental animals. CLSE images corresponded well with those of haematoxylin-eosin staining. CONCLUSIONS: CLSE on an ex vivo in vitro basis in experimental pigs is feasible.

[Quality of life of Crohn's disease patients]. / Kvalita zivota u Crohnovy nemoci.

BACKGROUND: The assessment of the quality of life in chronic diseases has started to move from clinical studies to practice. The main goal of the project was to assess the quality of life of Crohn's disease patients from two Czech centres by means of Czech versions of the general World Health Organization Quality of Life--BREF and specific Inflammatory Bowel Disease Questionnaire, to compare the quality of life of patients with an active disease and those in remission and to compare the quality of life with the general Czech population. METHODS AND RESULTS: 103 patients with Crohn's disease underwent a survey performed by means of two Czech versions of quality of life questionnaire. The dataset consisted of 53 men and 50 women; the average age of patients was 42 years. Increased activity was observed in 45 patients; 58 patients were in remission. By means of WHOQOL-BREF, we found the average global score of quality of life and satisfaction with health in our group to be 3.5 (Czech standard 3.8) and 2.8 (Czech standard 3.7), respectively. The results were compared to the Czech standards. A negative influence of disease activity was statistically significant (p < 0.001) in all domains using either of the two questionnaires. Clinical factors such as the use of corticosteroids or immunosuppressives, and the history of surgery influenced some domains. CONCLUSIONS: The results from our study indicate that the disease activity may have a significant impact on the quality of life in Crohn's disease patients. A combination of the general and specific questionnaire may identify factors that would otherwise remain unappreciated properly. The quality of life of patients in remission was comparable to that of the general Czech population.