Calcium imaging in intact mouse acinar cells in acute pancreas tissue slices.

The physiology and pathophysiology of the exocrine pancreas are in close connection to changes in intra-cellular Ca2+ concentration. Most of our knowledge is based on in vitro experiments on acinar cells or acini enzymatically isolated from their surroundings, which can alter their structure, physiology, and limit our understanding. Due to these limitations, the acute pancreas tissue slice technique was introduced almost two decades ago as a complementary approach to assess the morphology and physiology of both the endocrine and exocrine pancreas in a more conserved in situ setting. In this study, we extend previous work to functional multicellular calcium imaging on acinar cells in tissue slices. The viability and morphological characteristics of acinar cells within the tissue slice were assessed using the LIVE/DEAD assay, transmission electron microscopy, and immunofluorescence imaging. The main aim of our study was to characterize the responses of acinar cells to stimulation with acetylcholine and compare them with responses to cerulein in pancreatic tissue slices, with special emphasis on inter-cellular and inter-acinar heterogeneity and coupling. To this end, calcium imaging was performed employing confocal microscopy during stimulation with a wide range of acetylcholine concentrations and selected concentrations of cerulein. We show that various calcium oscillation parameters depend monotonically on the stimulus concentration and that the activity is rather well synchronized within acini, but not between acini. The acute pancreas tissue slice represents a viable and reliable experimental approach for the evaluation of both intra- and inter-cellular signaling characteristics of acinar cell calcium dynamics. It can be utilized to assess many cells simultaneously with a high spatiotemporal resolution, thus providing an efficient and high-yield platform for future studies of normal acinar cell biology, pathophysiology, and screening pharmacological substances

Inhibition of NHE-1 Increases Smoke-Induced Proliferative Activity of Barrett’s Esophageal Cell Line

Several clinical studies indicate that smoking predisposes its consumers to esophageal inflammatory and malignant diseases, but the cellular mechanism is not clear. Ion transporters protect esophageal epithelial cells by maintaining intracellular pH at normal levels. In this study, we hypothesized that smoking affects the function of ion transporters, thus playing a role in the development of smoking-induced esophageal diseases. Esophageal cell lines were treated with cigarettesmoke extract (CSE), and the viability and proliferation of the cells, as well as the activity, mRNA and protein expression of the Na(+)/H(+) exchanger-1 (NHE-1), were studied. NHE-1 expression was also investigated in human samples. For chronic treatment, guinea pigs were exposed to tobacco smoke, and NHE-1 activity was measured. Silencing of NHE-1 was performed by using specific siRNA. CSE treatment increased the activity and protein expression of NHE-1 in the metaplastic cells and decreased the rate of proliferation in a NHE-1-dependent manner. In contrast, CSE increased the proliferation of dysplastic cells independently of NHE-1. In the normal cells, the expression and activity of NHE-1 decreased due to in vitro and in vivo smoke exposure. Smoking enhances the function of NHE-1 in Barrett’s esophagus, and this is presumably a compensatory mechanism against this toxic agent

The Importance of Aquaporin 1 in Pancreatitis and Its Relation to the CFTR Cl- Channel

Aquaporins (AQPs) facilitate the transepithelial water flow involved in epithelial fluid secretion in numerous tissues;however, their function in the pancreas is less characterized. Acute pancreatitis (AP) is a serious disorder in which specific treatment is still not possible. Accumulating evidence indicate that decreased pancreatic ductal fluid secretion plays an essential role in AP;therefore, the aim of this study was to investigate the physiological and pathophysiological role of AQPs in the pancreas. Expression and localization of AQPs were investigated by real-time PCR and immunocytochemistry, whereas osmotic transmembrane water permeability was estimated by the dye dilution technique, in Capan-1 cells. The presence of AQP1 and CFTR in the mice and human pancreas were investigated by immunohistochemistry. Pancreatic ductal HCO3- and fluid secretion were studied on pancreatic ducts isolated from wild-type (WT) and AQP1 knock out (KO) mice using microfluorometry and videomicroscopy, respectively. In vivo pancreatic fluid secretion was estimated by magnetic resonance imaging. AP was induced by intraperitoneal injection of cerulein and disease severity was assessed by measuring biochemical and histological parameters. In the mice, the presence of AQP1 was detected throughout the whole plasma membrane of the ductal cells and its expression highly depends on the presence of CFTR Cl- channel. In contrast, the expression of AQP1 is mainly localized to the apical membrane of ductal cells in the human pancreas. Bile acid treatment dose- and time-dependently decreased mRNA and protein expression of AQP1 and reduced expression of this channel was also demonstrated in patients suffering from acute and chronic pancreatitis. HCO3- and fluid secretion significantly decreased in AQP1 KO versus WT mice and the absence of AQP1 also worsened the severity of pancreatitis. Our results suggest that AQP1 plays an essential role in pancreatic ductal fluid and HCO3- secretion and decreased expression of the channel alters fluid secretion which probably contribute to increased susceptibility of the pancreas to inflammation

Aktív sztrecsingtechnika csoportos alkalmazásának tapasztalatai = Experiences of Active Stretching Technique Application in Group Setting

Minden olyan lágyrészdiszfunkció, ami az ízület mozgásterjedelmének korlátozásával, az izomerő és állóképesség csökkenésével jár, az ízületek mobilitását behatároló tényezők kialakulásához fognak vezetni. Szerzők célja volt megvizsgálni fiatal, egészséges nők körében a contract-relax autosztrecsing eljárás csoportos formában való alkalmazásának eredményességét. A kutatásban 15 fő, női egyetemi hallgató (23,30 ±2,29 év) vett részt. Az állapotfelmérés során vizsgálták az alsó végtagi antigravitációs izmok megnyújthatóságát és erejét, valamint az állóképességet. A terápia során egy aktív sztrecsingtechnikát, a contract-relax technikát alkalmazták csoportos formában. A program 5 héten keresztül zajlott, hetente 3 alkalommal. Egy nyújtó kezelés 45 perces volt. Az adatok feldolgozása SPSS 24.0 statisztikai program segítségével zajlott. Az adatok bemutatása átlagban és szórásban történt, a változások mértékének értékeléséhez párosított t-próbát alkalmaztak p ≤ 0,05 szignifikanciaszinttel. A jelentősen csökkent állóképességi kategóriába tartozott a csoport 80%-a. Az antigravitációs izmok izomegyensúlyának összetett hibái és a dinamikus izomerőcsökke-nés mindhárom állóképességi kategóriában jelentős arányban mutatkozott meg. A mobilitás csökkenése kategóriánként változó volt. A nyújtóprogram után az állóképesség szignifikán-san javult (ingafutás teszt t.e. 23,73 ±14,69; t.u. 27,00 ±15,38; p < 0,001). A jelentős rövidülést hordozók aránya minden állóképességi kategóriában csökkent. A dinamikus izomerő szignifikáns növekedését mérték (m. quadriceps femoris jobb t.e. 39,27 ±10,84; t.u. 59,67 ±13,63; p < 0,001; m. quadriceps femoris bal t.e. 46,53 ±9,91; t.u. 63,07 ±12,85; p < 0,001) a statikusét azonban nem. A normál dinamikus izomerővel rendelkezők aránya minden kategóriában magas lett. Fiatal egészséges nők körében a jelen levő súlyos izomegyensúlyi hibák is eredményesen kezelhetők aktív autosztrecsing-technikával csoportos formában. Az eljárás könnyen tanítható, a résztvevők életkori és nemi elvárásaihoz ideálisan illeszthető. Any soft tissue dysfunction that occurs alongside the limitation of the joint range of motion, loss of muscle strength and endurance, will lead to the emergence of factors limiting joint mobility. The authors aimed to investigate the efficiency of the contract-relax autostretching method in a group setting, among young, healthy women. The study involved 15 female university students (23,30 ±2,29 years). During the health assessment the extensibility and strength of the lower limb antigravity muscles, as well as endurance, were tested. The therapy implemented an active stretching technique, the contract-relax technique, in a group form. The program lasted 5 weeks, 3 times per week. One stretching session was 45 minutes. Data were processed using SPSS 24.0 statistical software. Data were presented as mean and standard deviation, to assess the extent of the changes a paired t-test with a significance level of p ≤ 0,05. 80% of the group belonged to the significantly deteriorated endurance category. Compound defects in muscle balance of antigravity muscles and dynamic muscle strength loss were present in salient proportions in all three endurance categories. The decline of mobility varied by category. Stamina improved significantly after the stretching program (shuttle test b.t. 23.73 ±14.69; a.t. 27.00 ±15.38; p < 0.001). The proportion of individuals carrying substantial shortening decreased in every endurance category. The ratio of individuals with normal dynamic muscle strength grew high in every category. The significant rise of dynamic muscle strength was measured (m. quadriceps femoris right b.t. 39,27 ±10,840; a.t. 59,67 ±13,63; p < 0,001; m. quadriceps femoris left b.t. 46,53 ±9,91; a.t. 63,07 ±12,85; p < 0,001), however, the static was not measured. Among young, healthy women even severe muscle imbalances can be treated effectively with active autostretching techniques in a group setting. The method can be taught easily, it can adhere to the expecta-tions of the participants based on their age and gender

Mechanisms of Post-Pancreatitis Diabetes Mellitus and Cystic Fibrosis-Related Diabetes

Anatomical proximity and functional correlations between the exocrine and endocrine pancreas warrant reciprocal effects between the two parts. Inflammatory diseases of the exocrine pancreas, such as acute or chronic pancreatitis, or the presence of cystic fibrosis disrupt endocrine function, resulting in diabetes of the exocrine pancreas. Although novel mechanisms are being increasingly identified, the intra- and intercellular pathways regulating exocrine-endocrine interactions are still not fully understood, making the development of new and more effective therapies difficult. Therefore, this review sought to accumulate current knowledge regarding the pathogenesis of diabetes in acute and chronic pancreatitis, as well as cystic fibrosis

Mouse organoid culture is a suitable model to study esophageal ion transport mechanisms

Altered esophageal ion transport mechanisms play a key role in inflammatory and cancerous diseases of the esophagus, but epithelial ion processes have been less studied in the esophagus because of the lack of a suitable experimental model. In this study, we generated 3D esophageal organoids (EOs) from two different mouse strains and characterized the ion transport processes of the EOs. EOs form a cell-filled structure with a diameter of 250-300 µm and generated from epithelial stem cells as shown by FACS analysis. Using conventional PCR and immunostaining, the presence of Slc26a6 Cl-/HCO3- anion exchanger (AE), Na+/H+ exchanger (NHE), Na+/HCO3- cotransporter (NBC), cystic fibrosis transmembrane conductance regulator (CFTR) and anoctamin 1 Cl- channels were detected in EOs. Microfluorimetric techniques revealed high NHE, AE, and NBC activities, whereas that of CFTR was relatively low. In addition, inhibition of CFTR led to functional interactions between the major acid-base transporters and CFTR. We conclude that EOs provide a relevant and suitable model system for studying the ion transport mechanisms of esophageal epithelial cells, and they can be also used as preclinical tools to assess the effectiveness of novel therapeutic compounds in esophageal diseases associated with altered ion transport processes