Gil K

Telocytes: new insight into the pathogenesis of gallstone disease

The major mechanisms of gallstone formation include biliary cholesterol hypersecretion, supersaturation and crystallization, mucus hypersecretion, gel formation and bile stasis. Gallbladder hypomotility seems to be a key event that triggers the precipitation of cholesterol microcrystals from supersaturated lithogenic bile. Telocytes, a new type of interstitial cells, have been recently identified in many organs, including gallbladder. Considering telocyte functions, it is presumed that these cells might be involved in the signalling processes. The purpose of this study was to correlate the quantity of telocytes in the gallbladder with the lithogenicity of bile. Gallbladder specimens were collected from 24 patients who underwent elective laparoscopic cholecystectomy for symptomatic gallstone disease. The control group consisted of 25 consecutive patients who received elective treatment for pancreatic head tumours. Telocytes were visualized in paraffin sections of gallbladders with double immunofluorescence using primary antibodies against c‐Kit (anti‐CD117) and anti‐mast cell tryptase. Cholesterol, phospholipid and bile acid levels were measured in gallbladder bile. The number of telocytes in the gallbladder wall was significantly lower in the study group than that in the control group (3.03 ± 1.43 versus 6.34 ± 1.66 cell/field of view in the muscularis propria, P < 0.001) and correlated with a significant increase in the cholesterol saturation index. The glycocholic and taurocholic acid levels were significantly elevated in the control subjects compared with the study group. The results suggest that bile composition may play an important role in the reduction in telocytes density in the gallbladder.

Loss of gallbladder interstitial Cajal-like cells in patients with cholelithiasis

Background  Interstitial cells of Cajal (ICCs) play an important role in the regulation of gut motility. There is growing evidence that interstitial Cajal‐like cells (ICLCs) are present in the gallbladder wall. We hypothesize that changes in the density of ICLCs in the gallbladder wall may lead to the development of cholelithiasis due to the impairment of the gallbladder motility. The purpose of this study was to identify ICLCs in the gallbladders of patients with gallstones and to assess their densities.

Evidence of interstitial Cajal-like cells in human gallbladder.

The aim of this study was to assess the presence of interstitial Cajal-like cells (ICLCs) in the human gallbladder and to determine their distinctive characteristics on the basis of double immunohistochemical stain- ing. Gallbladder specimens were obtained from 30 patients subjected to elective laparoscopic cholecystectomy for symptomatic gallstone disease. Tissue samples were fixed in 4% phosphate-buffered paraformaldehyde, processed, embedded in paraffin, and, after sectioning, routinely stained with HE. Tissue antigens were re- trieved using the heat-induced epitope retrieval (HIER) method. For simultaneous visualisation of two anti- gens, an indirect double immunofluorescence procedure was applied. ICLCs were defined as CD117-immunop- ositive and tryptase-immunonegative objects. They were predominantly fusiform in shape with sparse branches that were visible in some sections. ICLCs were observed throughout the organ including the gallbladders fun- dus, body (corpus) and neck, being most numerous in the corpus. The ICLCs were detected almost exclusively within the muscularis propria and they were arranged parallel to smooth muscle cells. The following subpopula- tions of ICLCs were observed: ICLC-IM (intramuscular ICLCs) localised between smooth muscle fibres form- ing one muscle bundle; and ICLC-IB (interbundle ICLCs) localised within the connective tissue separating smooth muscle bundles. Thus, the presence of ICLCs in the human gallbladder was clearly identified, demon- strated by double immunohistochemistry which was found to be a reliable method for differentiating ICLCs from mast cells. (Folia Histochemica et Cytobiologica 2012, Vol. 50, No. 4, 581-585)

Exogenous Ghrelin Accelerates the Healing of Acetic Acid-Induced Colitis in Rats

Previous studies have shown that ghrelin reduces colonic inflammation induced by trinitrobenzene sulfonic acid and dextran sodium sulfate. In the present study we determined the effect of treatment with ghrelin on the course of acetic acid-induced colitis in rats. Rectal administration of 3% acetic acid solution led to induction of colitis in all animals. Damage of the colonic wall was accompanied by an increase in mucosal concentration of pro-inflammatory interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), as well mucosal activity of myeloperoxidase. Moreover, induction of colitis led to a reduction in colonic blood flow and DNA synthesis. Administration of ghrelin after induction of colitis led to faster regeneration of the colonic wall and reduction in colonic levels of IL-1β, TNF-α, and myeloperoxidase. In addition, treatment with ghrelin improved mucosal DNA synthesis and blood flow. Our study disclosed that ghrelin exhibits a strong anti-inflammatory and healing effect in acetic acid-induced colitis. Our current observation in association with previous findings that ghrelin exhibits curative effect in trinitrobenzene sulfonic acid- and dextran sodium sulfate-induced colitis suggest that therapeutic effect of ghrelin in the colon is universal and independent of the primary cause of colitis.

The use of nuclear morphometry for the prediction of survival in patients with advanced cancer of the larynx.

Abstract We examined retrospectively whether the quantitative morphometric analysis of nuclear shapes in patients with advanced cancer of the larynx could be used as a prognostic factor. In all, specimens were taken from 90 patients treated by surgery in the Department of Otolaryngology, Jagiellonian University, Cracow, Poland, between 1987 and 1988. The follow-up period was no shorter than 5 years. In the group examined there were 59 patients with T3 tumors and 31 with T4 tumors. A neck dissection was performed on one or both sides in each case. Metastases in regional lymph nodes were found in 26 patients. Histologic grading was assessed in all cases. Fourteen parameters of nuclear shape were studied using a computer-assisted system of image analysis. Morphometric data were compared with patients’ survival rates. The worse survival rates were found to be linked with a nuclear area (NA) ≥ 64.82 μm2 and its standard deviation (SDNA) ≥ 20.10 μm2, a nuclear perimeter (NP) ≥ 32.45 μm and its variation (SDNP) ≥ 4.77 μm, nuclear density (ND) ≥ 22 215.63 and its variation (SDND) ≥ 6930.85 and nuclear roundness (NR) ≥ 0.76. By using multivariate Cox regression analysis the SDND, presence of metastases in lymph nodes and low tumor differentiation were found to be independent prognostic factors. No statistically significant correlation was found between the parameters examined, lymph node status and tumor differentiation.

Is Urinary NGAL Determination Useful for Monitoring Kidney Function and Assessment of Cardiovascular Disease? A 12-Month Observation of Patients with Type 2 Diabetes

Background. Diabetic kidney disease (DKD) may start as glomerular or tubular damage. We assessed kidney function during one-year-long observation of patients with type 2 diabetes mellitus (T2DM) after initiation of nephroprotective treatment, with emphasis on the changes in urinary neutrophil gelatinase-associated lipocalin (uNGAL), and evaluated the association between tubular damage and cardiovascular complications of T2DM. Materials and Methods. Adult T2DM patients (55) were assessed initially and 30 patients after 1 year. Albumin and uNGAL and creatinine were measured in first morning urine. Albumin/creatinine (uACR) and uNGAL/creatinine (uNCR) ratios were calculated. Results. In logistic regression, both uACR above 30 mg/g and uNCR the median (21.3 μg/g) were associated with cardiovascular complications, independently of classical risk factors and diabetes duration. One year after initiation of treatment, a significant reduction in HbA1c was observed. BMI and lipid profiles did not change. Increase in serum creatinine and reduction in eGFR occurred, along with decrease in uNGAL and uNCR. Increasing uNCR and uACR were associated with higher control HbA1c. The increase in uNCR was more frequent in patients with hypertension. Conclusions. Better glycemic control in T2DM patients results in improved tubular function, as reflected by reduced uNCR and uNGAL. First morning urine uNGAL and uNCR may be useful to assess renal function and cardiovascular risk, along with albuminuria and eGFR.

Essential Role of Growth Hormone and IGF-1 in Therapeutic Effect of Ghrelin in the Course of Acetic Acid-Induced Colitis

Previous studies have shown that ghrelin exhibits a protective and therapeutic effect in the gut. The aim of the present study was to examine whether administration of ghrelin affects the course of acetic acid-induced colitis and to determine what is the role of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) in this effect. In sham-operated or hypophysectomized male Wistar rats, colitis was induced by enema with 1 mL of 3% solution of acetic acid. Saline or ghrelin (given at the dose of 8 nmol/kg/dose) was administered intraperitoneally twice a day. Seven days after colitis induction, rats were anesthetized and the severity of the colitis was assessed. Treatment with ghrelin reduced the area of colonic mucosa damage in pituitary-intact rat. This effect was associated with increase in serum levels of GH and IGF-1. Moreover, administration of ghrelin improved blood flow in colonic mucosa and mucosal cell proliferation, as well as reduced mucosal concentration of proinflammatory interleukin-1β (IL-1β) and activity of myeloperoxidase. Hypophysectomy reduced serum levels of GH and IGF-1 and increased the area of colonic damage in rats with colitis. These effects were associated with additional reduction in mucosal blood follow and DNA synthesis when compared to pituitary-intact rats. Mucosal concentration of IL-1β and mucosal activity of myeloperoxidase were maximally increased. Moreover, in hypophysectomized rats, administration of ghrelin failed to affect serum levels of GH or IGF-1, as well as the healing rate of colitis, mucosal cell proliferation, and mucosal concentration of IL-1β, or activity of myeloperoxidase. We conclude that administration of ghrelin accelerates the healing of the acetic acid-induced colitis. Therapeutic effect of ghrelin in experimental colitis is mainly mediated by the release of endogenous growth hormone and IGF-1.

Serum Concentrations of Angiopoietin-2 and Soluble fms-Like Tyrosine Kinase 1 (sFlt-1) Are Associated with Coagulopathy among Patients with Acute Pancreatitis

In severe acute pancreatitis (SAP), systemic inflammation leads to endothelial dysfunction and activation of coagulation. Thrombotic disorders in acute pancreatitis (AP) include disseminated intravascular coagulation (DIC). Recently, angiopoietin-2 and soluble fms-like tyrosine kinase 1 (sFlt-1) were proposed as markers of endothelial dysfunction in acute states. Our aim was to assess the frequency of coagulation abnormalities in the early phase of AP and evaluate the relationships between serum angiopoietin-2 and sFlt-1 and severity of coagulopathy. Sixty-nine adult patients with AP were recruited: five with SAP, 15 with moderately severe AP (MSAP) and 49 with mild AP. Six patients were diagnosed with DIC according to International Society on Thrombosis and Haemostasis (ISTH) score. All patients had at least one abnormal result of routine tests of hemostasis (low platelet count, prolonged clotting times, decreased fibrinogen, and increased D-dimer). The severity of coagulopathy correlated with AP severity according to 2012 Atlanta criteria, bedside index of severity in AP and duration of hospital stay. D-dimers correlated independently with C-reactive protein and studied markers of endothelial dysfunction. Angiopoietin-2, D-dimer, and ISTH score were best predictors of SAP, while sFlt-1 was good predictor of MSAP plus SAP. In clinical practice, routine tests of hemostasis may assist prognosis of AP.

Telocytes in female reproductive system (human and animal).

Telocytes (TCs) are a newly discovered type of cell with numerous functions. They have been found in a large variety of organs: heart (endo‐, myo‐, epi‐ and pericardium, myocardial sleeves, heart valves); digestive tract and annex glands (oesophagus, stomach, duodenum, jejunum, liver, gallbladder, salivary gland, exocrine pancreas); respiratory system (trachea and lungs); urinary system (kidney, renal pelvis, ureters, bladder, urethra); female reproductive system (uterus, Fallopian tube, placenta, mammary gland); vasculature (blood vessels, thoracic duct); serous membranes (mesentery and pleura); and other organs (skeletal muscle, meninges and choroid plexus, neuromuscular spindles, fascia lata, skin, eye, prostate, bone marrow). Likewise, TCs are widely distributed in vertebrates (fish, reptiles, birds, mammals, including human). This review summarizes particular features of TCs in the female reproductive system, emphasizing their involvement in physiological and pathophysiological processes.

Interstitial Cajal-Like Cells and Bile Lithogenicity in the Pathogenesis of Gall-Stone Disease

UNLABELLED Gall-stone disease constitutes a serious clinical problem and is the most frequent cause of elective cholecystectomies. There are many etiopatogenic factors however; lithogenic bile and its stasis due to gall-bladder hypomotility seem to be the most important. In recent years discovery of pacemaker function of Interstitial Cells of Cajal changed our understanding of smooth muscle physiology and helped to disclose many gastrointestinal motility disorders. THE AIM OF THE STUDY was identification and quantification of interstitial Cajal-like cells (ICLCs) in gall-bladder muscle wall from patients with cholelithiasis and in gall-stone-free controls, as well as determination of the relationship between the number of ICLCs and Cholesterol Saturation Index (CSI) of bile in both analyzed groups. MATERIAL AND METHODS 20 patients operated for symptomatic cholelithiasis were enrolled into the study group. The control group consisted of 20 patients operated for pancreatic head tumors, with no pre- and intraoperative signs of gall-stones. Identification of ICLCs in the gall-bladder was performed by means of double immunofluorescence technique with anti c-Kit and anti-mast cell tryptase antibodies. Quantitative analysis was carried out under fluorescence microscopy conjoined with image analysis software. Bile samples were used for calculation of CSI. RESULTS ICLCs were detected within gall-bladder muscle wall. Number of ICLCs was statistically significantly lower in patients from the study group as compared to control. The study also revealed statistically significantly higher CSI in the study group. CONCLUSIONS The quantity of ICLCs is diminished in the gall-bladder from patients with cholelithiasis and there is negative correlation between the number of ICLCs and CSI of bile. Regarding the role of ICCs in regulation of GI tract motility, it appears that reduction in their number may be important etiopatogenic factor of cholelithiasis.