The Role of Galectin-1 and Galectin-3 in the Mucosal Immune Response to Citrobacter rodentium Infection

Despite their abundance at gastrointestinal sites, little is known about the role of galectins in gut immune responses. We have therefore investigated the Citrobacter rodentium model of colonic infection and inflammation in Galectin-1 or Galectin-3 null mice. Gal-3 null mice showed a slight delay in colonisation after inoculation with C. rodentium and a slight delay in resolution of infection, associated with delayed T cell, macrophage and dendritic cell infiltration into the gut mucosa. However, Gal-1 null mice also demonstrated reduced T cell and macrophage responses to infection. Despite the reduced T cell and macrophage response in Gal-1 null mice, there was no effect on C. rodentium infection kinetics and pathology. Overall, Gal-1 and Gal-3 play only a minor role in immunity to a gut bacterial pathogen

Galectin-4 Controls Intestinal Inflammation by Selective Regulation of Peripheral and Mucosal T Cell Apoptosis and Cell Cycle

Galectin-4 is a carbohydrate-binding protein belonging to the galectin family. Here we provide novel evidence that galectin-4 is selectively expressed and secreted by intestinal epithelial cells and binds potently to activated peripheral and mucosal lamina propria T-cells at the CD3 epitope. The carbohydrate-dependent binding of galectin-4 at the CD3 epitope is fully functional and inhibited T cell activation, cycling and expansion. Galectin-4 induced apoptosis of activated peripheral and mucosal lamina propria T cells via calpain-, but not caspase-dependent, pathways. Providing further evidence for its important role in regulating T cell function, galectin-4 blockade by antisense oligonucleotides reduced TNF-alpha inhibitor induced T cell death. Furthermore, in T cells, galectin-4 reduced pro-inflammatory cytokine secretion including IL-17. In a model of experimental colitis, galectin-4 ameliorated mucosal inflammation, induced apoptosis of mucosal T-cells and decreased the secretion of pro-inflammatory cytokines. Our results show that galectin-4 plays a unique role in the intestine and assign a novel role of this protein in controlling intestinal inflammation by a selective induction of T cell apoptosis and cell cycle restriction. Conclusively, after defining its biological role, we propose Galectin-4 is a novel anti-inflammatory agent that could be therapeutically effective in diseases with a disturbed T cell expansion and apoptosis such as inflammatory bowel disease

Epac1 mediates protein kinase A–independent mechanism of forskolin-activated intestinal chloride secretion

Intestinal Cl− secretion is stimulated by cyclic AMP (cAMP) and intracellular calcium ([Ca2+]i). Recent studies show that protein kinase A (PKA) and the exchange protein directly activated by cAMP (Epac) are downstream targets of cAMP. Therefore, we tested whether both PKA and Epac are involved in forskolin (FSK)/cAMP-stimulated Cl− secretion. Human intestinal T84 cells and mouse small intestine were used for short circuit current (Isc) measurement in response to agonist-stimulated Cl− secretion. FSK-stimulated Cl− secretion was completely inhibited by the additive effects of the PKA inhibitor, H89 (1 µM), and the [Ca2+]i chelator, 1,2-bis-(o-aminophenoxy)-ethane-N,N,N’,N’-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM; 25 µM). Both FSK and the Epac activator 8-pCPT-2’-O-Me-cAMP (50 µM) elevated [Ca2+]i, activated Ras-related protein 2, and induced Cl− secretion in intact or basolateral membrane–permeabilized T84 cells and mouse ileal sheets. The effects of 8-pCPT-2’-O-Me-cAMP were completely abolished by BAPTA-AM, but not by H89. In contrast, T84 cells with silenced Epac1 had a reduced Isc response to FSK, and this response was completely inhibited by H89, but not by the phospholipase C inhibitor U73122 or BAPTA-AM. The stimulatory effect of 8-pCPT-2’-O-Me-cAMP on Cl− secretion was not abolished by cystic fibrosis transmembrane conductance (CFTR) inhibitor 172 or glibenclamide, suggesting that CFTR channels are not involved. This was confirmed by lack of effect of 8-pCPT-2’-O-Me-cAMP on whole cell patch clamp recordings of CFTR currents in Chinese hamster ovary cells transiently expressing the human CFTR channel. Furthermore, biophysical characterization of the Epac1-dependent Cl− conductance of T84 cells mounted in Ussing chambers suggested that this conductance was hyperpolarization activated, inwardly rectifying, and displayed a Cl−>Br−>I− permeability sequence. These results led us to conclude that the Epac-Rap-PLC-[Ca2+]i signaling pathway is involved in cAMP-stimulated Cl− secretion, which is carried by a novel, previously undescribed Cl− channel

Gene expression profiling in sinonasal adenocarcinoma

<p>Abstract</p> <p>Background</p> <p>Sinonasal adenocarcinomas are uncommon tumors which develop in the ethmoid sinus after exposure to wood dust. Although the etiology of these tumors is well defined, very little is known about their molecular basis and no diagnostic tool exists for their early detection in high-risk workers.</p> <p>Methods</p> <p>To identify genes involved in this disease, we performed gene expression profiling using cancer-dedicated microarrays, on nine matched samples of sinonasal adenocarcinomas and non-tumor sinusal tissue. Microarray results were validated by quantitative RT-PCR and immunohistochemistry on two additional sets of tumors.</p> <p>Results</p> <p>Among the genes with significant differential expression we selected <it>LGALS4, ACS5, CLU, SRI and CCT5 </it>for further exploration. The overexpression of <it>LGALS4, ACS5, SRI</it>, <it>CCT5 </it>and the downregulation of <it>CLU </it>were confirmed by quantitative RT-PCR. Immunohistochemistry was performed for LGALS4 (Galectin 4), ACS5 (Acyl-CoA synthetase) and CLU (Clusterin) proteins: LGALS4 was highly up-regulated, particularly in the most differentiated tumors, while CLU was lost in all tumors. The expression of ACS5, was more heterogeneous and no correlation was observed with the tumor type.</p> <p>Conclusion</p> <p>Within our microarray study in sinonasal adenocarcinoma we identified two proteins, LGALS4 and CLU, that were significantly differentially expressed in tumors compared to normal tissue. A further evaluation on a new set of tissues, including precancerous stages and low grade tumors, is necessary to evaluate the possibility of using them as diagnostic markers.</p

Evaluation of nutritive value of fodders from permanent grasslands in organic farms

Badania prowadzono w latach 2004-2007 w 34 ekologicznych gospodarstwach łąkarskich prowadzących produkcję zwierzęcą. W ramach badań analizowano próbki zielonek z łąk i pastwisk, próbki siana oraz kiszonek z runi łąkowej. Oceniano zawartość wybranych składników mineralnych (N, P, K, Ca, Mg i Na) i składników pokarmowych (popiół surowy, białko ogólne, włókno surowe i tłuszcz surowy). Wartość energetyczną pasz wyrażono w MJ oraz wg systemu INRA - w JPM i JPŻ. Zarówno zawartość składników pokarmowych jak i wartość energetyczna analizowanych pasz średnio mieściły się w granicach wartości uznawanych za optymalne. Stwierdzono jednakże bardzo duże ich wahania, szczególnie w odniesieniu do białka ogólnego i włókna surowego. Średnia zawartość fosforu była zadowalająca w zielonce pastwiskowej oraz nieznacznie niedoborowa w pozostałych paszach. Średnia zawartośćpotasu i magnezu była zadowalająca we wszystkich rodzajach pasz. Zawartość wapnia była minimalnie niedoborowa tylko w sianie, natomiast zawartość sodu wyjątkowo mała we wszystkich rodzajach pasz. W porównaniu z wynikami badań siana z gospodarstw konwencjonalnych w Polsce w badaniach w gospodarstwach ekologicznych stwierdzono większy udziałpróbek o większej zawartości fosforu, wapnia i magnezu, o zbliżonym udziale potasu i znacznie mniejszej zawartości sodu. W celu lepszego wykorzystania pasz z trwałych użytków zielonych i poprawy ich jakości, konieczne jest doskonalenie metod użytkowania łąk, w tym gospodarki nawozowej, oraz technologii sporządzania pasz, szczególnie w gospodarstwach o dużym udziale TUZ, w których produkcja zwierzęca stanowi podstawowe źródło ich dochodu.The studies in 34 organic grassland farms with livestock production were carried out in the years 2004-2007. Samples of meadow sward, pasture green fodder, hay and grass silage were analysed in the study. The content of mineral (N, P, K, Ca, Ma and Na) and nutritive components (crude ash, total protein, crude fibre and crude fat) was analysed. Energetic value of forages was expressed in MJ and according to the INRA system – in UFL and UVF. Both the content of nutritive components and the energetic value of analysed fodders fell within the values considered the optimum. However, very large variation was found of these values, especially in total protein and crude fibre contents. The average content of phosphorus in pasture green fodder was satisfactory and slightly deficient in other fodders. The average content of potassium and magnesium in all kinds of fodders was satisfactory. The calcium content was slightly deficient only in hay, while the sodium content was exceptionally low in all kinds of fodders. More hay samples from organic farms showed higher content of phosphorus, calcium and magnesium, similar content of potassium and considerably smaller content of sodium than hay samples from conventional farms in Poland. For the purpose of better utilisation of fodders from permanent grasslands and improvement of their quality, it is necessary to improve the methods of meadows utilisation, including fertilisation, and technologies of fodders production, especially in farms with large share of permanent grasslands, where animal production determines the basic source of income

Nitrogen, phosphorus and potassium balances in ecological farms of differential areas

Badania ankietowe przeprowadzone w wybranych łąkarskich gospodarstwach ekologicznych w siedmiu województwach naszego kraju umożliwiła dokonanie oceny gospodarki azotem, fosforem i potasem poprzez bilanse tych składników. Salda bilansowe były zróżnicowane w poszczególnych grupach obszarowych. Najbardziej korzystne salda bilansu azotu i fosforu oraz znaczne wyrównanie sald potasu, zarówno w 2005 oraz 2006 r. stwierdzono w grupie obszarowej gospodarstw od 10,1 do 20,0 ha. Gospodarstwa w tej grupie charakteryzowały się dużą obsadą zwierząt, ok. 0,8 DJPźha-1, mniejszym udziałem użytków zielonych w UR oraz dużą różnorodnością upraw polowych na gruntach ornych. Najwyższe salda bilansowe potasu (najwyższe nadmiar), stwierdzono w 2006 r. w grupie gospodarstw najmniejszych (1,0-10,0 ha), o jednocześnie największej obsadzie zwierząt gospodarskich. Gospodarstwa z grup obszarowych od 20,1 do 50,0 ha i >50 ha charakteryzowały się mniejszą obsadą zwierząt, dominującym udziałem zbóż w strukturze zasiewów oraz najbardziej niekorzystnymi saldami bilansowymi azotu i fosforu oraz potasu, głównie w 2005 r.Selected ecological grassland farms located in seven Polish provinces were inquired to make possible to evaluate the nitrogen, phosphorus and potassium management through balances of these nutrients. Balances in particular area groups were clearly varied. The most profitable nitrogen and phosphorus balances with considerable equal potassium balances were noticed in 10.1-20.0 ha area group both in 2005 and 2006. Farms in this group had high livestock density, about 0.8 LUźha-1, lower share of grasslands in croplands and high diversity of arable farming. The highest balances of potassium (the highest surpluses) were noticed in 2006 in 1.0-10.0 ha area group with the highest livestock density. The farms of 20.1-50.0 ha and > 50 ha area groups had lower livestock density, with greater cereals share in cropping system and the most unprofitable nitrogen, phosphorus and potassium balances, especially in 2005

Analysis of nitrogen, phosphorus and potassium balance in meadow organic farms

Ekologiczny sposób gospodarowania ogranicza ilość składników wprowadzanych do gospodarstwa z zewnątrz i może zakłócać/utrudniać racjonalną gospodarkę tymi składnikami. Oceny gospodarowania składnikami N, P, K w wybranych, zróżnicowanych obszarowo, gospodarstwach ekologicznych z dużym udziałem trwałych użytków zielonych dokonano na podstawie badań ankietowych prowadzonych w latach 2005-2006. Opracowano bilanse tych składników wykorzystując metodę "u wrót gospodarstwa". Salda tych bilansów były wyraźnie zróżnicowane w zależności od obszaru gospodarstwa, a także obsady zwierząt, struktury zasiewów i udziału trwałych użytków zielonych w użytkach rolnych i udziału bobowatych w runi. Najwyższe salda bilansu azotu (nadwyżki), stwierdzono w grupie gospodarstw najmniejszych 1,0-10,0 ha, charakteryzujących się największą obsadą zwierząt gospodarskich, i największych, z grupy >50 ha o znacznie mniejszej obsadzie zwierząt i z dominacją zbóż w strukturze zasiewów. Najbardziej wyrównane salda bilansu azotu wystąpiły w grupie gospodarstw 10,1-20,0 ha, charakteryzujących się średnią obsadą zwierząt (0,8 DJP·ha-¹), mniejszym udziałem użytków zielonych w UR oraz dużą różnorodnością upraw polowych. Salda bilansu fosforu i potasu w większości gospodarstw były ujemne, wskazujące przede wszystkim na potrzebę zwiększenia obsady zwierząt i produkcji obornika.Ecological way of farming may limit the input of elements to the farm and may disturb/hamper rational use of these elements. N, P and K management in selected organic farms of various areas and large part of permanent grasslands was assessed based on questionnaire studies in the years 2005-2006. Nutrient balance was calculated with the "at the farm's gate" method. The balances were distinctly differentiated depending on farms' area, animal stock, crop structure, percentage share of grasslands in total croplands and the share of legumes in sward. The largest nitrogen surplus was found in smallest farms of an area from 1.0 to 10.0 ha having large animal stock and in the largest farms of an area over 50 ha with much smaller animal stock and cereals dominating in the crop structure. Most even nitrogen balance was characteristic for farms of 10.1 to 20.0 ha with medium animal stock (0.8 LU ha-¹), smaller percentage of grasslands and variable field crops. Phosphorus and potassium balance was negative in most farms indicating the need for increasing animal stock and manure production

Corrective factors of intestinal microflora disorders in the perinatal period

Intestinal microflora is found at approximately 10⁴ bacteria per ml of intestinal fluid. Although this ecosystem is very diverse with dynamic changes taking place within it, there is a certain stability in the type and number of species, referred to as the core microbiome, found in 95% of the human population. Genetic variation of microorganisms is relatively small, and their functions are strictly defined and highly conservative. The microbiome exists symbiotically with the host, protecting it against colonization by pathogenic microorganisms, providing essential metabolites, and stimulating the immune system. Colonization begins prenatally and its development is greatly influenced by the course of pregnancy, method of delivery, food supplied to the child during the first moments of life, and post-birth environment. The appropriate intestinal microflora composition is a key determinant of health and homeostasis, and any intestinal dysbiosis can be associated later in life with the development of obesity, diabetes, allergies or cancer. Due to the increasing number of hospitalised pregnancies and deliveries, affecting the intestinal microflora of a newborn, efforts are being made to minimize this process and restore the newborn’s microbiome. The use of the Vaginal seeding procedure raises up great hopes, but also some fears concerning its safety. Some very simple and most natural factors have been recently also appreciated and promoted, such as breastfeeding or direct contact of the baby’s skin with the mother’s skin, which are allies to probiotic bacteria. The purpose of this paper is to emphasize the importance of microbial colonization of the human body and to present the latest and most effective procedures that are designed to correct the existing dysbiosis or to reduce the risk of its occurrence. The literature for the compilation of this study has been obtained from databases such as PubMed, Google Scholar, Web of Science