Localization of TFF1 and TFF3 proteins at different stages of the mouse embryonic development

Cilj istraživanja: TFF proteini uključeni su u razne fiziološke procese u probavnom, dišnom, mokraćnom i živčanom sustavu. Osim toga, prisutni su i u patološkim stanjima. Utječu na migraciju, diferencijaciju i apoptozu stanica, kao i na angiogenezu i imunološki odgovor te se intenzivno istražuju u zdravlju i bolesti. S obzirom da ima vrlo malo podataka o ekspresiji TFF proteina tijekom embrionalnog razvoja, u ovom radu imunohistokemijskom metodom istražena je prisutnost TFF1 i TFF3 proteina u tkivima mišjih zametaka starosti od 14 do 18 dana. Analizirani su i histomorfološki parametri epifizne ploče rasta i sekundarnih centara okoštavanja na proksimalnom dijelu tibije divljeg tipa i miševa s isključenim genom TFF3 starih mjesec dana. Materijal i metode: Imunohistokemijski pokusi izvedeni su na preparatima 23 zametka CD1 miševa fiksiranih 4%-nim paraformaldehidom i uklopljenih u parafin, starosti od 14 do 18 dana. Histomorfometrijski parametri analizirani su na tibijama miševa divljeg tipa visoko srođenih životinja razvijenih križanjem sojeva 129/Sv i C57BL/6j te miševa s isključenim genom TFF3. Korišteno je 5 miševa po skupini, a histološki preparati tibija obojeni su trikromnim bojenjem po Massonu. Rezultati: Prisutnost TFF 1 i 3 proteina u mišjim zamecima uočena je u sluznici probavnog i dišnog sustava, gušterači, kanalićima bubrega, epidermisu kože, živčanom sustavu te okoštavanju, a pojavljivala se ovisno o stadiju razvoja miša i stupnju diferencijacije tkiva. Smještaj TFF1 i TFF3 proteina u tkivima mišjih zametaka nije bio povezan s podrijetlom tkiva prema embrionalnim zametnim listićima. Nisu pronađene značajne histomorfološke razlike u debljini epifizne ploče rasta, pojedinih zona enhondralnog okoštavanja niti u gustoći hondrocita po jedinici površine između divljeg tipa miševa i miševa s isključenim genom TFF3, međutim, u sekundarnim centrima okoštavanja u epifizama mišjih tibija u u životinja s isključenim genom TFF3 vrijednosti gustoće volumena, gustoće površine i broja koštanih gredica bile su znatno manje, a razdvojenost koštanih gredica znatno veća. Zaključak: TFF1 i TFF3 protein zbog svojih brojnih fizioloških uloga važni su u razvoju zametka te u nekim slučajevima mogu poslužiti kao markeri fiziološkog sazrijevanja tkiva. TFF3 vjerojatno ima važan učinak na stvaranje i kvalitetu spužvaste kosti u sekundarnim centrima okoštavanja.Objectives: TFF proteins are involved in different physiological processes in the digestive, respiratory, urinary and nervous system and are also present in different pathological conditions. These proteins influence cell migration, differentiation and apoptosis, as well as angiogenesis and immune response and are under active research in health and disease. Since very limited data are available on the expression of TFF proteins in the embryonic tissues, presence of TFF1 and TFF3 in the tissues of 14 to 18-day old mice was investigated in this work. Also, histomorphological parameters of the epiphyseal growth plate and of the secondary ossification centers were investigated in the proximal tibia of the wild type and TFF3 knock-out mice. Material and methods: Immunohistochemical experiments were done manually, using 23 formalin fixed, paraffin embedded CD1 mouse embryos, 14 to 18-day old. Histomorphometric parameters were analyzed in the tibiae of inbred wild-type mice developed by crossing 129/Sv and C57BL/6j strain and TFF3 knock-out mice. 5 mice per group were used, and histological sections of tibiae were stained using Masson’s trichrome stain. Results: Presence of TFF1 and TFF3 proteins was detected in the gastrointestinal and respiratory mucosa, pancreas, kidney tubules, epidermis, nervous system and areas of ossification of mouse embryos and occurred depending on the stage of embryonic development and tissue differentiation. Localization of TFF1 and TFF3 protein in mouse embryonic tissues didn’t depend on the germinal layer from which the tissue originated. No significant histomorphological differences between wild-type mice and TFF3 knock out mice were found in epiphyseal plate thickness, in the thickness of different zones of endochondral ossification, and in the chondrocyte density. However, cancellous bone in the secondary ossification centers of mouse tibia epiphyses showed significantly lower values of volume density, surface density and trabecular number, while trabecular separation was significantly higher. Conclusion: Because of their numerous physiological roles, TFF1 and TFF3 are important in the embryonic development, and in some cases may serve as early markers of tissue maturation. TFF3 probably has an important effect on the formation and quality of the cancellous bone in the secondary ossification centers

E Actitrode: The new selective stimulation interface for functional movements in hemiplegics patients

We describe the new multi-contact electrode-array for surface electrical stimulation, and the corresponding interface device that allows on-line selection of the conductive fields during the application of the system. This new device has a specific value for therapeutic applications of electrical stimulation since it allows effective generation of desired functional movements. The user-friendly interface also allows patients at home to select the optimal electrode array; thereby, to receive therapies out of the clinical environment. The electrode was tested in three post-stroke hemiplegics patients. The pilot experiments showed that system works sufficiently good for control of fingers during grasp and release functions without the interference of the wrist movement. The use of electrode is also envisioned for many other applications (foot-drop fitness, shoulder subluxation, etc)

Raspodjela rizičnih čimbenika za moždani udar u istočnoj Hrvatskoj

The aim of this study was to determine the distribution of risk factors according to age, gender, subtypes and recurrence of stroke in eastern Croatia. The study included 250 acute stroke patients admitted to University Department of Neurology, Osijek University Hospital Centre in 2011. Patients were grouped according to age, gender, subtypes and recurrence of stroke. The study showed significant differences in the distribution of cigarette smoking, diabetes, cardiomyopathy and hyperuricemia according to patient age. According to gender, male patients had a significantly higher prevalence of smoking and alcohol abuse, whereas in female patients the prevalence of arterial hypertension, atrial fibrillation and hyperuricemia was significantly higher. Regarding stroke subtypes, significant differences were noticed in the prevalence of arterial hypertension, atrial fibrillation, cardiomyopathy and cerebral blood vessel stenosis. Atrial fibrillation was significantly more common in first-ever than in recurrent stroke. Study results identified the groups of patients requiring special attention regarding particular risk factors in eastern Croatia and emphasized the need of developing regional strategies of screening, prevention and holistic care for stroke patients.Cilj ovoga istraživanja bio je odrediti raspodjelu rizičnih čimbenika u bolesnika s moždanim udarom u istočnoj Hrvatskoj prema dobi, spolu, podtipovima i ponovnom javljanju. U istraživanje bilo je uključeno 250 bolesnika koji su pretrpjeli akutni moždani udar i primljeni su na Kliniku za neurologiju Kliničkoga bolničkog centra Osijek u 2011. godini. Bolesnici su podijeljeni prema dobi, spolu, podtipovima i ponovnom javljanju. Istraživanje je pokazalo značajne razlike u raspodjeli pušenja cigareta, dijabetesa, kardiomiopatije i hiperuricemije prema dobi bolesnika. Prema spolu, učestalost pušenja i zlouporabe alkohola bila je značajno veća kod muških bolesnika, dok je učestalost arterijske hipertenzije, atrijske fibrilacije i hiperuricemije bila značajno veća kod žena. Prema podtipovima moždanog udara, značajne razlike uočene su kod učestalosti arterijske hipertenzije, atrijske fibrilacije, kardiomiopatije i stenoze cerebralnih krvnih žila. Atrijska fibrilacija bila je značajno češća u prvom nego u ponovljenom moždanom udaru. Naši rezultati pokazuju koje skupine bolesnika u istočnoj Hrvatskoj zahtijevaju posebnu pozornost s obzirom na pojedine rizične čimbenike te naglašavaju potrebu osmišljavanja regionalnih strategija za probir, prevenciju i holističku brigu za oboljele od moždanog udara

The use of honey in contemporary wound treatment

Med je prirodan pčelinji proizvod složenoga kemijskoga sastava i zanimljivih fizikalnih i bioloških svojstava. Premda sadrži mikroorganizme u dormantnom stanju, odlikuje se snažnim antimikrobnim učinkom. Suvremena medicina sve obilnije koristi antimikrobna svojstva meda u terapiji rana različite etiologije, na primjer inficiranih kirurških rana i kroničnih venskih ulkusa. Sve brojnija klinička istraživanja izvještavaju o uspješnom korištenju medicinskih pripravaka meda u liječenju inficiranih rana kod kojih je prethodno neuspješno korištena antibiotska terapija. Premda su potrebna dodatna istraživanja, sasvim je jasno da će zbog odličnih rezultata, ekonomičnosti i zadovoljstva pacijenata uporaba meda u liječenju rana zauzimati sve istaknutije mjesto u medicinskoj praksi.Honey is a natural bee product, with complex chemical composition and interesting physical and biological properties. Although it contains dormant microbes, honey has strong antimicrobial effect. This effect is increasingly used in modem medicine for treating wounds of versatile etiology, for example infected surgical wounds and chronic venous ulcers. Numerous clinical trials have reported successful usage of medical-grade honey for treating infected wounds in which antibiotic therapy had previously been unsuccessful. Though additional research is needed, it is clear that the use of honey will take notable place in medical practice due to its excellent results, patient’s satisfaction and for being economic

Trefoil factor family protein 3 affects cancellous bone formation in the secondary centers of ossification of mouse tibiae

Background and Purpose: Trefoil factor family (TFF) is a small family of peptides, which comprises three peptides, TFF1, TFF2 and TFF3. The primary sites of TFF expression and synthesis are mucous epithelia, with gastrointestinal mucosa being their main localization. They have a role in mucosal restitution and strengthening of the normal mucosal barrier. Although not present in healthy articular cartilage, TFF3 is expressed in osteoarthritis and septic arthritis affected cartilage and promotes cartilage degradation in such conditions. Also, TFF3 is present in endochondral ossification during embryonic development. Since this implies a role for TFF3 in the ossification process, we evaluated the histomorphometric parameters of the trabecular bone in the epiphyses of mouse tibiae from wild type mice and TFF3 knock-out mice. Materials and Methods: Tibiae of wild-type and knock-out mice were isolated, fixed in 4% paraformaldehyde, paraffin embedded and cut into 6μm sections, which were stained using Masson’s trichrome stain. Digital photographs were taken for histomorphometry of the epiphyseal cancellous bone. Results: Trabecular bone volume density, trabecular bone surface density and trabecular number were significantly decreased in TFF3 knock-out mice, when compared to wild-type mice. Trabecular separation was significantly higher in TFF3 knock-out mice, and trabecular thickness did not differ significantly. Conclusions: In addition to its impact on the cartilage degradation, our present study shows that TFF3 might also have a role in the formation of cancellous bone and its properties

Trefoil factor family protein 3 affects cancellous bone formation in the secondary centers of ossification of mouse tibiae

Background and Purpose: Trefoil factor family (TFF) is a small family of peptides, which comprises three peptides, TFF1, TFF2 and TFF3. The primary sites of TFF expression and synthesis are mucous epithelia, with gastrointestinal mucosa being their main localization. They have a role in mucosal restitution and strengthening of the normal mucosal barrier. Although not present in healthy articular cartilage, TFF3 is expressed in osteoarthritis and septic arthritis affected cartilage and promotes cartilage degradation in such conditions. Also, TFF3 is present in endochondral ossification during embryonic development. Since this implies a role for TFF3 in the ossification process, we evaluated the histomorphometric parameters of the trabecular bone in the epiphyses of mouse tibiae from wild type mice and TFF3 knock-out mice. Materials and Methods: Tibiae of wild-type and knock-out mice were isolated, fixed in 4% paraformaldehyde, paraffin embedded and cut into 6μm sections, which were stained using Masson’s trichrome stain. Digital photographs were taken for histomorphometry of the epiphyseal cancellous bone. Results: Trabecular bone volume density, trabecular bone surface density and trabecular number were significantly decreased in TFF3 knock-out mice, when compared to wild-type mice. Trabecular separation was significantly higher in TFF3 knock-out mice, and trabecular thickness did not differ significantly. Conclusions: In addition to its impact on the cartilage degradation, our present study shows that TFF3 might also have a role in the formation of cancellous bone and its properties

Localization of trefoil factor family peptide 3 (TFF3) in epithelial tissues originating from the three germ layers of developing mouse embryo

Trefoil factor family (TFF) peptides are involved in the maintenance of epithelial integrity and epithelial restitution. Mature epithelial tissues originate from different embryonic germ layers. The objective of this research was to explore the presence and localization of TFF3 peptide in mouse embryonic epithelia and to examine if the occurrence of TFF3 peptide is germ layer-dependent. Mouse embryos (14-18 days old) were fixed in 4% paraformaldehyde and embedded in paraffin. Immunohistochemistry was performed with affinity purified rabbit anti-TFF3 antibody, goat anti-rabbit biotinylated secondary antibody and streptavidin-horseradish peroxidase, followed by 3, 3’-diaminobenzidine. TFF3 peptide was present in the gastric and intestinal mucosa, respiratory mucosa in the upper and lower airways, pancreas, kidney tubules, epidermis, and oral cavity. The presence and localization of TFF3 peptide was associated with the embryonic stage and tissue differentiation. TFF3 peptide distribution specific to the germ layers was not observed. The role of TFF3 peptide in cell migration and differentiation, immune response, and apoptosis might be associated with specific embryonic epithelial cells. TFF3 peptide may also be considered as a marker for mucosal maturation

Pojavnost intramiokardijalnih masnih stanica u stijenci desne pretklijetke i desne klijetke – postmortalna humana analiza

Histologic and radiologic studies describe intramyocardial fat tissue as a normal finding or as part of cardiac pathology. The role of fat cells within the myocardium is not fully understood. The aim of this study was to assess fat tissue distribution in the myocardium of right atrium (RA) and right ventricle (RV) and age differences in subjects free from cardiac disease. The study included 10 males without cardiac disease divided into two groups according to age (below/above 50 years). Three cross sections were performed (RV free wall and apex and RA free wall) with histomorphological analysis on digital photographs. The shares of total myocardial fat (TMF), perivascular fat (PVF) and non-perivascular (nPVF) fat were calculated. Samples from the older group had larger amounts of fat in the epicardium and myocardium, without statistically significant difference (TMF p=0.847, PVF p=0.4 and nPVF p=0.4). The largest quantities of fat tissue were found in the RV apex samples (14.9%), followed by RV free wall (7.5%) and RA (4.5%), where total apical RV fat share was significantly larger than in RA sample (p=0.044). Intramyocardial fat cells were present within the non-diseased RA and RV in all samples, mostly in the apex. Further investigations on age difference, effect of visceral obesity and sex differences are needed.Dosadašnja histološka i radiološka istraživanja opisuju pojavnost intramiokardijalnog masnog tkiva kao dio normalnog nalaza ili kao dio patološkog nalaza srca. Uloga intramiokardijalnih masnih stanica nije još razjašnjena. Cilj ovoga istraživanja bio je utvrditi raspodjelu masnog tkiva u miokardu desne pretklijetke (DP) i desne klijetke (DK) ovisno o dobi i lokalizaciji kod ispitanika bez kardijalnog morbiditeta. Patohistološka postmortalna studija uključila je 10 muškaraca bez kardijalnih bolesti podijeljenih u dvije skupine prema dobi (ispod/iznad 50 godina). Uzorci su uzeti u tri presjeka (projekcija slobodnog zida DK, slobodni zid vrha DK te stijenka DP) s histomorfološkom analizom na digitalnim fotografijama. Izračunati su udjeli ukupnog masnog tkiva unutar srčanog mišića (UMT), perivaskularnog masnog tkiva (PMT) i ne-perivaskularnog masnog tkiva (nPMT). U uzorcima starijih ispitanika prisutna je veća količina masnog tkiva u epikardu i miokardu, bez statistički značajne razlike (UMT p=0,847, PMT p=0,4, nPMT p=0,4). Najveće količine masnog tkiva pronađene su u uzorcima vrha DK (14,9%), nakon čega slijedi stijenka slobodnog zida DK (7,5%) i DP (4,5%), gdje je ukupni udio masnog tkiva DK bio znatno veći nego kod uzorka DP (p=0,044). Intramiokardijalne masne stanice sastavni su dio histološkog nalaza DP i DK u svim uzorcima bez prisutnih kardijalnih bolesti, dominantno u vrhu DK. Neophodna su daljnja istraživanja utjecaja dobi, spola i visceralne pretilosti na pojavnost intramiokardijalnih masnih stanica unutar stijenke desnog srca

Floristic composition of permanent grassland in the nature park Stara Planina (Serbia)

The five most important meadow associations on the Stara Planina Mountain in National Park "Stara Planina" were analysed in order to define initial parameters for determination of grassland potential, as well as natural resources for production of biologically valuable and high quality food and revitalization of agricultural production in hilly-mountainous region. Floristic composition of five meadow associations is presented: Medicago falcate - Festucetum rubrae at three locations around the Pirot, Triflolio - Agrostietum stoloniferae Lj. Mark. 1973, Agrostietum vulgaris Z. Pavl. 1955, two ass. Festucetum vallesiaceae - Agrostietum vulgaris Danon et Blaz. 1978 and Festuco-Agrostietum Horv. (1951) 1982.em Trinajest.1972 one. All five associations are around Dimitrovgrad. First association of Festucetum vallesiaceae - Agrostietum vulgaris Danon et Blaz. 1978 had the highest diversity with the total number of 75 species. The number of species in investigated associations was: grasses from 8 to 23, legumes from 8 to 17 and other species from 21 to 34. Participation of major meadow species based on their mass is presented in the paper. On the basis of their productivity the species belong to the associations of useful grasses, useful legumes and other useful species and weeds. The study of grasslands was done in order to establish production potentials and quality of grasslands used for grazing of small ruminants, i.e. sheep and goats

The effect of nitrogen fertilization on quality and yield of grass-legume mixtures

Studies were carried out in pure stands of three grasses: cocksfoot, meadow fescue and perennial ryegrass, and lucerne, as well as their mixtures with different ratios of individual components. The trial was carried out on an experimental field of the Institute for Animal Husbandry (Belgrade, Serbia). Fertilization with N caused a significant increase of yield. The highest yields were determined in treatments with 100 kg N ha(-1), and the lowest in treatments without fertilization. Fertilization increased significantly the CP content, but without impact on other quality parameters. CP, NDF and ADF differed significantly in various crops