Molecular cloning, polymorphism, and expression analysis of the LKB1/STK11 gene and its association with non-specific digestive disorder in rabbits.

Liver kinase B1 (LKB1, also called STK11) encodes a serine/threonine kinase mutated in Peutz-Jeghers cancer syndrome characterized by gastrointestinal polyposis. Although LKB1 plays an important role in regulating energy homeostasis, cell growth, and metabolism via activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK), nothing is known about its molecular characteristics and possible involvement in non-specific digestive disorder (NSDD) of rabbits. In the present study, we first cloned the coding sequence (CDS) of rabbit LKB1, which consisted of 1317 bp encoding 438 amino acids (AAs) and contained a highly conserved S_TKc kinase domain. Its deduced AA sequence showed 87.93-91.10% similarities with that of other species. In order to determine its involvement in NSDD, a NSDD rabbit model was built by a dietary fiber deficiency. The polymorphic site of LKB1 was then investigated in both healthy and NSDD groups using directing sequencing. Our results suggested that a synonymous variant site (840 c. G > C, CCC→CCG) existed in its S_TKc domain, which was associated with susceptibility to NSDD. Furthermore, qPCR was utilized to examine the mRNA levels of LKB1 and its downstream targets (i.e., PRKAA2, mTOR and NF-kß) in several intestinal-related tissues from both healthy and NSDD groups. Significant changes in their expression levels between two groups indicated that impaired LKB1 signaling contributed to the intestinal abnormality in NSDD rabbits. Taken together, it could be concluded that LKB1 might be a potential candidate gene affecting the occurrence of rabbit NSDD. This information may serve as a basis for further investigations on rabbit digestive diseases.

[Antibodies to enzymes of purine metabolism as a factor of gastrointestinal tract lesions in systemic scleroderma].

Antibodies to Adenosine Deaminase (AD) and Guanin Deaminase (GDA) were founded in sera of systemic sclerosis (SS) patients, their concentration positive correlated with activity of disease. Increased levels of antibodies to AD and GDA correlated with the severity of gastrointestinal tract injury (especially pancreatitis and hepatitis). It is shown that level of antibodies to AD and GDA differs reliable from contents of these antibodies, in sera of healthy persons. It is shown that AD enzymatic activity was decreased, while activity of GDA was increased. The antibodies to enzymes may be one of the possible causes of change of enzymes activity in sera of SS patients. The method of immunoenzyme determination of level of antibodies to AD, G on the basis of immobilized form of magnet sorbent was developed for immune diagnostics of SS.

Nox enzymes and oxidative stress in the immunopathology of the gastrointestinal tract.

Chronic inflammation caused by Helicobacter pylori infection or inflammatory bowel disease (IBD) is closely linked to cancer development. Innate immune abnormalities and enhanced production of reactive oxygen species through a phagocyte NADPH oxidase (Nox2) are key issues in understanding the pathogenesis of inflammation-dependent carcinogenesis. Besides Nox2, functionally distinct homologues (Nox1, Nox3, Nox4, Nox5, Duox1, and Duox2) have been identified. Nox1 and Duox2 are highly expressed in the gastrointestinal tract. Although the functional roles of Nox/Duox in the gastrointestinal tract are still unclear, we will review their potential roles in the gastrointestinal immunopathology, particularly in H. pylori-induced inflammation, IBD, and malignancy.

Anti-inflammatory drugs in the 21st century.

Historically, anti-inflammatory drugs had their origins in the serendipitous discovery of certain plants and their extracts being applied for the relief of pain, fever and inflammation. When salicylates were discovered in the mid-19th century to be the active components of Willow Spp., this enabled these compounds to be synthesized and from this, acetyl-salicylic acid or Aspirin was developed. Likewise, the chemical advances of the 19th-20th centuries lead to development of the non-steroidal anti-inflammatory drugs (NSAIDs), most of which were initially organic acids, but later non-acidic compounds were discovered. There were two periods of NSAID drug discovery post-World War 2, the period up to the 1970's which was the pre-prostaglandin period and thereafter up to the latter part of the last century in which their effects on prostaglandin production formed part of the screening in the drug-discovery process. Those drugs developed up to the 1980-late 90's were largely discovered empirically following screening for anti-inflammatory, analgesic and antipyretic activities in laboratory animal models. Some were successfully developed that showed low incidence of gastro-intestinal (GI) side effects (the principal adverse reaction seen with NSAIDs) than seen with their predecessors (e.g. aspirin, indomethacin, phenylbutazone); the GI reactions being detected and screened out in animal assays. In the 1990's an important discovery was made from elegant molecular and cellular biological studies that there are two cyclo-oxygenase (COX) enzyme systems controlling the production of prostanoids [prostaglandins (PGs) and thromboxane (TxA2)]; COX-1 that produces PGs and TxA2 that regulate gastrointestinal, renal, vascular and other physiological functions, and COX-2 that regulates production of PGs involved in inflammation, pain and fever. The stage was set in the 1990's for the discovery and development of drugs to selectively control COX-2 and spare the COX-1 that is central to physiological processes whose inhibition was considered a major factor in development of adverse reactions, including those in the GI tract. At the turn of this century, there was enormous commercial development following the introduction of two new highly selective COX-2 inhibitors, known as coxibs (celecoxib and rofecoxib) which were claimed to have low GI side effects. While found to have fulfilled these aims in part, an alarming turn of events took place in the late 2004 period when rofecoxib was withdrawn worldwide because of serious cardiovascular events and other coxibs were subsequently suspected to have this adverse reaction, although to a varying degree. Major efforts are currently underway to discover why cardiovascular reactions took place with coxibs, identify safer coxibs, as well as elucidate the roles of COX-2 and COX-1 in cardiovascular diseases and stroke in the hope that there may be some basis for developing newer agents (e.g. nitric oxide-donating NSAIDs) to control these conditions. The discovery of the COX isoforms led to establishing their importance in many non-arthritic or non-pain states where there is an inflammatory component to pathogenesis, including cancer, Alzheimer's and other neurodegenerative diseases. The applications of NSAIDs and the coxibs in the prevention and treatment of these conditions as well as aspirin and other analogues in the prevention of thrombo-embolic diseases now constitute one of the major therapeutic developments of the this century. Moreover, new anti-inflammatory drugs are being discovered and developed based on their effects on signal transduction and as anti-cytokine agents and these drugs are now being heralded as the new therapies to control those diseases where cytokines and other nonprostaglandin components of chronic inflammatory and neurodegenerative diseases are manifest. To a lesser extent safer application of corticosteroids and the applications of novel drug delivery systems for use with these drugs as well as with NSAIDs also represent newer technological developments of the 21st century. What started out as drugs to control inflammation, pain and fever in the last two centuries now has exploded to reveal an enormous range and type of anti-inflammatory agents and discovery of new therapeutic targets to treat a whole range of conditions that were never hitherto envisaged.

[Role of nitric oxide (NO) in the etiopathogenesis of certain alimentary diseases].

Nitric oxide is one of essential regulators of the physiological functions of the gastrointestinal tract including vascular tone, transfer of nerve impulses, protection of mucous coats, motor activity and secretion. The review of literature presents data on sources of nitric oxide, its molecular and cellular interaction, role in the genesis of different diseases of the gastrointestinal tract. The experimental and clinical material provides grounds for approaches to the regulation of this signaling pathway with the help of nitric oxide donors, activators and inhibitors of its formation, arginine, antioxidants, phosphodiesterase and guanylate cyclase inhibitors. The bibliography includes 93 references.

[Digestive pathology and digestive function of small intestine mucous layer]. / Pishchevaritel'naia funktsiia pristenochnogo slizistogo sloia tonkoi kishki pri patologii organov pishchevareniia.

The research of small intestine mucous layer at children and adults is carried out. It is revealed high meanings enzyme distribution in human small intestine mucous layer and their change at celiac disease, food allergy, short-bowel syndrome.

Different patterns in immunoreactive anionic and cationic trypsinogen in urine and serum in human acute pancreatitis.

BACKGROUND: Acute pancreatitis (AP) results in elevated concentrations of trypsinogen (T) isoenzymes in serum. Immunoreactive anionic trypsinogen in urin (irAT/u) is elevated in AP, and has recently been proposed as a rapid diagnostic instrument and severity predictor. These results have not been confirmed by other groups, and irAT/u has not been further characterized. The concentration of immunoreactive cationic trypsinogen in urine (irCT/u) and the serum irAT/irCT ratio in AP have not been extensively examined. METHODS: Levels of irAT and irCT were studied in urine and serum from 50 AP patients and in urine from 41 non-AP patients. Severity was assessed according to the Atlanta classification. irAT/u was characterized by gel filtration. RESULTS: Gel filtration revealed only AT in the urine. Highly significant differences in irAT/u were seen between AP/non-AP (p < 0.0001) and mild/severe disease (p = 0.0012). The irAT/irCT ratio in serum changed from normal 0.8 to 1.3 in AP. CONCLUSIONS: IrAT and only traces of irCT were found in the urine in AP. IrAT/u was higher in AP than in other acute abdominal disorders (non-AP) and also higher in severe than in mild AP. IrAT in serum (irAT/s) increased proportionally more than irCT/s in AP, but did not discriminate mild from severe forms. High levels of irAT/u in some non-AP cases and a wide range in AP cases make the clinical value of the test questionable.

[Metabolic disorders in digestive and hepatobiliary pathology in those who worked in the cleanup of the aftermath of the accident at the Chernobyl Atomic Electric Power Station]. / Narusheniia metabolizma pri digestivnoi i gepatobiliarnoi patologii u likvidatorov posledstvii avarii na ChAES.

A clinical observation over 57 liquidators of the Chernobyl accident aftermath (ChAAL) with associated diseases of the digestive and hepato-biliary system revealed abnormalities in blood enzymic activity, presenting as elevation of, in particular, alanine aminotransferase as well as of asparagine aminotransferase, gamma-glutaminetranspeptidase, alkaline phosphatase, lactate dehydrogenase, and of blood mineral composition at the expense of an increase in certain minerals. Disturbances in underlying process of bodily metabolism promote the development and association of erosive and ulcerous lesions of the stomach and duodenum with chronic noncalculous cholecystitis, persistent hepatitis in ChAAL.

Expression of pancreatic digestive enzymes in normal and pathologic epithelial cells of the human gastrointestinal system.

Pancreatic digestive enzymes have rarely been reported in human nonpancreatic organs. We examined their expression in the epithelial cells of the nonpancreatic gastrointestinal organs, looking for pancreatic alpha-amylase, trypsin, chymotrypsin and pancreatic lipase. Western blotting, enzyme assay and pancreatic alpha-amylase mRNA were also used in selected specimens. In normal tissues, immunoreactivity of one or more of these enzymes was frequently noted in cells of the salivary glands, stomach, duodenum, large pancreatic ducts, extrahepatic bile ducts and gall bladder. The epithelium of the normal oesophagus, small intestine and colon were consistently negative for these enzymes. In pathologic tissues, immunoreactivity for one or more enzymes was present in epithelial cells of pleomorphic adenomas of the salivary glands, oesophageal squamous cell carcinoma, gastric adenoma and adenocarcinoma, pancreatic adenocarcinoma, cholecystitis, adenocarcinoma of the gall bladder and extrahepatic bile duct, and colon adenoma and adenocarcinoma. Western blotting showed a specific band of each enzyme in some specimens of normal stomach. In situ hybridization for pancreatic alpha-amylase mRNA showed specific signals in the normal stomach, but not in the normal colon. Reverse transcriptase polymerase chain reaction analysis for pancreatic alpha-amylase mRNA revealed specific signals in the normal stomach. Enzyme assay revealed that the stomach and gall bladder showed these activities. The data suggest that pancreatic digestive enzymes are produced by several epithelial cell types of the nonpancreatic gastrointestinal organs, that the organs positive for pancreatic enzyme have a common cell lineage, and that neoplasms continue to express or neoexpress these enzymes after neoplastic transformation.

Serum amylase and lipase in the evaluation of acute abdominal pain.

The purpose of this study was to determine 1) the incidence and magnitude of elevation in admission serum amylase and lipase levels in extrapancreatic etiologies of acute abdominal pain, and 2) the test most closely associated with the diagnosis of acute pancreatitis. Serum amylase and lipase levels were obtained in 306 patients admitted for evaluation of acute abdominal pain. Patients were categorized by anatomic location of identified pathology. Logistic regression analysis was used to compare the enzyme levels between patient groups and to determine the correlation between elevation in serum amylase and lipase. Twenty-seven (13%) of 208 patients with an extrapancreatic etiology of acute abdominal pain demonstrated an elevated admission serum amylase level with a maximum value of 385 units (U)/L (normal range 30-110 U/L). Twenty-six (12.5%) of these 208 patients had an elevated admission serum lipase value with a maximum of 3685 U/L (normal range 5-208 U/L). Of 48 patients with abdominal pain resulting from acute pancreatitis, admission serum amylase ranged from 30 to 7680 U/L and lipase ranged from 5 to 90,654 U/L. Both serum amylase and lipase elevations were positively associated with a correct diagnosis of acute pancreatitis (P < 0.001) with diagnostic efficiencies of 91 and 94 per cent, respectively. A close correlation between elevation of admission serum amylase and lipase was observed (r = 0.87) in both extrapancreatic and pancreatic disease processes. Serum amylase and lipase levels may be elevated in nonpancreatic disease processes of the abdomen. Significant elevations (greater than three times upper limit of normal) in either enzyme are uncommon in these disorders. The strong correlation between elevations in the two serum enzymes in both pancreatic and extrapancreatic etiologies of abdominal pain makes them redundant measures. Serum lipase is a better test than serum amylase either to exclude or to support a diagnosis of acute pancreatitis.

Serum trypsinogen-2 and trypsin-2-alpha(1)-antitrypsin complex in malignant and benign digestive-tract diseases. Preferential elevation in patients with cholangiocarcinomas.

Serum concentrations of trypsinogen-2 and trypsin-2-alpha(1)-antitrypsin (trypsin-2-AAT) were determined in 145 patients with malignant and 61 with benign digestive-tract diseases. The validity of these tests for detection of cancer was compared with that of CA 19-9 and CEA. Elevated levels of trypsinogen-2 (>90 micrograms/l) and trypsin-2-AAT (>25 micrograms/l) were found in 46% and 42%, respectively, of patients with malignant disease and the levels of trypsinogen-2 were significantly higher than in those with benign disease (p<0.005). High trypsinogen-2 and trypsin-2-AAT concentrations were found most often in patients with biliary and pancreatic cancer, but also in benign obstructive biliary disease. Our results suggest that trypsinogen-2 and trypsin-2-AAT are new potential markers for cholangiocarcinomas.

Quantitative and qualitative changes of extracellular-superoxide dismutase in patients with various diseases.

Extracellular-superoxide dismutase (EC-SOD) is a secretory glycoprotein that is the major SOD isozyme in extracellular fluids. It has previously been shown that EC-SOD levels in sera from healthy persons are clearly divided into two discontinuous groups: a lower group (named Group I, below 120 ng/ml) and a higher group (Group II, above 400 ng/ml). The family studies have shown that the high EC-SOD level in healthy persons is genetically transmitted. We report here on the EC-SOD levels in the sera of patients with various diseases. The EC-SOD levels were distinctly higher in patients with renal diseases and moderately higher in liver diseases and diabetes than those in normal healthy persons. In cerebrovascular diseases, heart diseases and acute digestive diseases, significant differences of EC-SOD were not observed. In patients with renal diseases, the increase of EC-SOD was accompanied by the lack of renal function. Serum EC-SOD in Group I healthy persons is known to be heterogeneous with regard to heparin affinity and can be separated into three fractions: A without affinity, B with weak affinity and C with relatively strong heparin affinity, whereas the EC-SOD in Group II is mainly one fraction of C-type. Also in the case of hemodialysis patients, serum EC-SOD in Group I or Group I' (approximately 120-400 ng/ml) was divided into three fractions. EC-SOD in Group II showed two different profiles on heparin-Sepharose column chromatographies: one consisted mainly of EC-SOD C and the other consisted of EC-SOD A and C. It is probable that the high serum EC-SOD level in hemodialysis patients was due to two possible factors: the genetic transmitted factor and unknown pathophysiological factor(s).

Glutathione S-transferase activity in patients with cancer of the digestive tract.

Glutathione S-transferase (GST) and carcinoembryonic antigen were measured in the plasma of 95 patients with neoplasm of digestive tract, in 40 patients suffering from non-neoplastic diseases and in 40 healthy subjects. The mean value of the GST activity was significantly (P < 0.001) elevated in patients with gastric, liver and colorectal cancer (10.4 U/l, 14.1 U/l and 12.3 U/l respectively) as compared with the reference population (3.2 U/l). GST elevations above normal were observed in 26 (90%) patients with gastric cancer, in 18 (100%) with liver cancer and in 25 (89%) with colorectal cancer. Carcinoembryonic antigen appeared less sensitive. In 15 patients the postoperative levels of serum GST were increased after surgery then gradually declined and after 1 month showed a normalization in 10 patients. Our data suggest that GST measurement may be useful as a tumour marker in gastric, liver and colorectal cancer. Moreover the combined determination of GST and other markers increase the sensitivity for cancer detection.

[The formation of a hypokinetic syndrome of the digestive system under conditions of weightlessness]. / Formirovanie gipokineticheskogo sindroma pishchevaritel'noi sistemy v usloviiakh nevesomosti.

In space flights of various duration, the gastropancreatic complex turned out to be the most liable part of the digestive system. The main changes occurring in the gastrointestinal tract involve an increase in the peptic gastric potential, pancreatic hyperenzymemia, change of the glycemic curve forms, a decrease in the liver and pancreas density.

[The regulation of hydrolytic processes in the small and large intestines]. / O reguliatsii gidroliticheskikh protsessov v tonkoi i tolstoi kishkakh.

A study is presented of the effect of different modifiers (analogues of natural food) on the rate of starch hydrolysis in the small and large intestine. Revealed were different changes of the activity of amylase and glycyl-l-leucin-dipeptidase under the effect of B vitamins, Enzaprost F., cimetidine, obsidan, prednisolone, dopamine, ethaden, ethanol. Both in vitro and in vivo studies showed that the reactions of changes of the enzymatic activity were of the same direction.

Paneth cell degranulation and lysozyme secretion during acute equine alimentary laminitis.

The equine Paneth cell response to a shift in the microbial balance of the intestinal tract was studied by inducing an acute episode of alimentary laminitis in 6 mature ponies. The normal bacterial population of the gut was modified by administration of a carbohydrate-rich ration. During acute laminitis a dramatic degranulation of the Paneth cells occurred in the intestinal glands throughout the duodenum, jejunum, and ileum. Bacteriocidal lysozyme, which was immunohistochemically identified as a component of the Paneth cell secretory granule, was evident in the glandular lumina and in degranulated Paneth cells. These results indicate that lysozyme is secreted by the equine Paneth cell in an apparent attempt to regulate the changing microbial population induced by carbohydrate overload of the gut. From these observations, it is suggested that the Paneth cell plays a role in the mucosal defense system of the equine intestinal tract.

[Limited proteolysis activation syndrome in surgery]. / Sindrom aktivatsii ogranichennogo proteoliza v khirurgii.

Under consideration is an original author's conception, formulated previously, of the syndrome of activation of limited proteolysis as an obligatory pathogenetic link of all pathological processes including the alteration at the tissue level as applied to surgical diseases. A universal character of the syndrome in acute and chronic surgical diseases and traumas, its role in their pathogenesis was shown in 1107 patients and 130 experimental animals. Variants of the course of the syndrome in different diseases are considered both in the postoperative and posttraumatic periods. The significance of the syndrome of activation of limited proteolysis for practical surgery is discussed.