Inštitut za predklinične vede

Vpliv taninov na žleze slinavke pri merjaščkih

lukam — Mon, 21 Nov 2022 07:45:15 +0000

Influence of tannin dietary inclusion on boar salivary glands lukam Mon, 11/21/2022 - 08:57 Inštitut za predklinične vede Prašiči Prehranski dodatki tanini podušesna slinska žleza imunohistokemija histologija Research

Salivary gland adaptation to dietary inclusion of hydrolysable tannins in boars.

Tannins have traditionally been avoided in animal nutrition due to their anti-nutritive effects however, recent studies have reported beneficial antimutagenic, anticancerogenic, antidiarrheal and antiulcerogenic effects of tannins on animals. In a comprehensive study investigating the effect of dietary tannins in feed on boar taint in entire males, a significant enlargement of the parotid salivary gland (parotidomegaly) was observed. In contrast, mass of the mandibular glands showed no significant changes between the experimental groups. Pigs were divided into four groups: control group (T0) and three experimental groups fed with 1% (T1), 2% (T2) or 3% (T3) of tannins. In our study, we have performed a histological analysis of salivary glands to determine the morphological and immunohistochemical properties leading to parotidomegaly. We have observed the enlargement of acinar area and of the nucleus to cytoplasm ratio in the serous cells in T3 group. Moreover, we have observed a linear trend towards enlargement of lobular area, acinar area and the number of ducts per lobule in all three experimental groups. Additionally, by immunohistochemistry, using antibodies against acid and basic proline rich proteins (PRPs), we have shown significantly higher amounts of PRPs in parotid glands of pigs fed with high tannin concentrations (T3 group). Consistent with the effect of tannins on mandibular gland mass, we did not detect any significant morphological changes. Therefore, the results of our study demonstrate the importance of increased functional activity of the parotid salivary glands as the first and most important line of defence against high dietary tannin concentrations and their potential negative effects.

Authors: Maša Mavri, Marjeta Čandek-Potokar, Gregor Fazarinc, Martin Škrlep, Catrin S. Rutland, Božidar Potočnik, Nina Batorek-Lukač in Valentina Kubale.

Original article published in https://www.mdpi.com/2076-2615/12/17/2171/pdf?version=1661342653

Srčna kost - pregledni članek

lukam — Mon, 26 Sep 2022 08:02:58 +0000

Review of os cordis lukam Mon, 09/26/2022 - 10:15 Inštitut za predklinične vede srčna kost osifikacija ektopična kost Research

Anatomy, histology, morphology, development and functions of os cordis: A review

Figure: Relative position of Ossa cordis in the heart, dorsal aspect with of ventriculi cordis with the atria removed (drawing by Samo Krivec, Veterinary faculty, University of Ljubljana; adopted from Habel et al., 2018, Illustrated Nomina Anatomica veterinaria ( 4th ed.).

Anatomy, histology, morphology, development and functions of os cordis: A review

Systematic review regarding heart skeleton published in Anatomia Histologia Embriologia journal, highlights the similarities and variations in ossa cordis prevalence, morphology, histology and anatomical location between differing veterinary species and in humans. It also identifies associated factors such as aging and cardiovascular disease for each species in relation to functional roles and developmental mechanisms that these bone structures may play. The potential functions of os cordis are presented, ranging from aiding cardiac contraction and conduction, providing cardiac structure, and protecting components of the heart, through to counteracting high mechanical stress. Furthermore, this review discusses the evidence and rationale behind the theories regarding the formation and development of ossa cordis in different veterinary species and in people.

The article is also very useful in pedagogical work with students. Samo Krivec, a student at the Veterinary faculty, University in Ljubljana, participated in the article as the author of one of the figures, which was drawn according to the NAV model (Nomina anatomica veterinaria illustrated) exclusively for this article.

Authors: Adam Best, Monika Egerbacher, Sophia Swaine, William Pérez, Aziza Alibhai, Paul Rutland, Valentina Kubale, Samir A.A. El-Gendy, Mohamed, A.M. Alsafy, Kerstin Baiker, Craig J Sturrock, and Catrin Sian Rutland

Original article published in: https://onlinelibrary.wiley.com/doi/10.1111/ahe.12861

Imunokastracija odraslih merjascev

lukam — Thu, 28 Jul 2022 05:14:09 +0000

Immunocastration in adult boars lukam Thu, 07/28/2022 - 07:20 Inštitut za predklinične vede Prašiči imunokastracija protitelesa proti GnRH morfometrija mod qPCR Research

Immunocastration in adult boars as a model for late-onset hypogonadism.

While immunocastration has been studied in male pre-pubertal pigs, data on older, sexually mature animals are limited. To understand the physiological effects of androgen deprivation in the late sexual development phase, we compared mature immunocastrated boars (n = 19) to young male immunocastrated pigs (n = 6) and young entire males (n = 6) as positive and negative controls, respectively. We hypothesized that the timing of gonadotropin-releasing hormone suppression (early or late sexual development phases) influences the extent of reproductive function inhibition, histological structure of testicular tissue, and expression levels of selected genes related to steroid metabolism. Antibody titer, hormonal status, and histomorphometric analysis of testicular tissue were subjected to principal component analysis followed by hierarchical clustering. In mature post-pubertal boars immunocastration resulted in progressive but variable regression of testicular function. We hypothesize that this is due to local activation of the GnRH2-GnRHR2 system in the testes, which is independent of the hypothalamic-pituitary-gonadal axis. Further research is needed to undoubtedly confirm this hypothesis.

This study contributes significantly to the understanding of the effects of immunocastration in adult boars at physiological and cellular levels, while serving as an animal model for late hypogonadism in translational medicine. We have also identified parameters that could be used to evaluate the success of immunocastration in boars.

Authors: dr. Nina Batorek Lukač, dr. Kevin Kress, dr. Marjeta Čandek-Potokar, dr. Gregor Fazarinc, dr. Martin Škrlep, Klavdija Poklukar, dr. Raffael Wesoly, dr. Volker Stefanski in dr. Milka Vrecl

Original article published in: https://onlinelibrary.wiley.com/doi/full/10.1111/andr.13219

Molekulska opredelitev receptorskih ortologov BILF1

lukam — Fri, 10 Jun 2022 11:32:03 +0000

Molecular characterization of BILF1 receptor orthologues lukam Fri, 06/10/2022 - 13:38 Inštitut za predklinične vede Epstein-Barr virus porcine lymphotropic herpesvirus BILF1 G protein signaling; MHC class I drug target post-transplant lymphoproliferative disease in vivo model Research

Epstein-Barr virus-encoded BILF1 orthologues from porcine lymphotropic herpesviruses display common molecular functionality.

Epstein-Barr virus (EBV; γ-herpesvirus) is an oncovirus, that is prevalent in approximately 95% of the world’s population and is a driving factor of numerous, mainly lymphoproliferative diseases. Despite the importance of EBV as a pathogenic and etiologic agent, there is still no vaccine or antiviral drug against the virus. One reason for this is that the virus has evolved many different strategies to avoid host immune system. One of these mechanisms is the expression of a viral G protein coupled receptor (vGPCR) BILF1 in the infected cells. vGPCRs are receptors that have been adopted by the host through years of coevolution and modified by the virus to serve its survival. vGPCRs also mimic the functional properties of endogenously expressed GPCRs. BILF1 encoded in EBV (EBV-BILF1) is known for its ability to downregulate surface expressed major histocompatibility complex I molecules (MHC-I) at the cell surface and cell transformation through constitutive Gαi activity. In addition to EBV, BILF1 is also encoded in three porcine γ-herpesviruses named porcine lymphotropic herpesviruses 1-3 (PLHV1-3). Based on the importance of PLHV1 in the development of post-transplant lymphoproliferative disease (PTLD), which resembles morphologic and histologic features of EBV driven human disease, pigs infected with PLHV1 were proposed as a model to study this disease and EBV driven pathogenesis. Lately, vGPCRs are drawing attention as drug targets for antiviral drugs based on their exclusive expression on infected cells and thereby possibility for specific targeting. The proposed porcine model would therefore allow the investigation of BILF1 as a drug target, which requires an in depth pharmacological characterization of the receptor. In our study, we have focused on characterization of BILF1 orthologues from all four viruses (EBV, PLHV1, PLHV2 in PLHV3) and comparison of their cellular localization, constitutive signaling, constitutive internalization and the ability to avoid host immune system. Using state of the art methodology, we have observed that these properties are conserved in all BILF1 orthologues. Some discrepancy was found in the ability to activate downstream transcription factors NFAT and NF-κB, which play important roles in tumour formation by other vGPCRs. Importantly, with our study we have shown progressive expression of PLHV1-BILF1 in tissue samples from PTLD diseased pigs, suggesting the importance of the receptor in the PTLD aetiology.

Original article published in: https://www.frontiersin.org/articles/10.3389/fendo.2022.862940/full

Authors: Maša Mavri, Valentina Kubale, Daniel P. Depledge, Jianmin Zuo, Christene A. Huang, Judith Breuer, Milka Vrecl, Michael A. Jarvis, Eva Jarc Jovičić, Toni Petan, Bernhard Ehlers, Mette M. Rosenkilde, Katja Spiess

Srečanje partnerjev projekta "¨BestCHEESE" v Turčiji

lukam — Thu, 07 Oct 2021 07:29:39 +0000

The meeting of the international project "BestCHEESE" in Turkey lukam Thu, 10/07/2021 - 09:43 Inštitut za predklinične vede Erasmus+ Turčija BestCHEESE Education

From 20 to 24 September 2021, an international meeting of the project "Investigation of Traditional Cheese Production Technology and Standards in the Light of European Union - BestCHEESE" was held in Turkey within the framework of the Erasmus+ programme and under the auspices of the Eskisehir Agriculture and Forestry Directorate.

The aim of the project is to standardize the production of dairy products by developing national hygiene and production standards.

Beside the Veterinary Faculty of the University in Ljubljana, research institutions from Turkey (Eskisehir Agriculture and Forestry Directorate, Ankara University Faculty of Veterinary Medicine, Batman Agriculture and Forestry Directorate), Italy (Azienda Sanitaria Provinciale di Agrigento) and Slovakia (Slovak University of Agriculture in Nitra) are participating in the project. For Slovenia, Tilen Vake, DVM and Neža Žnidaršič, DVM from the Veterinary Faculty participated in the meeting.

The meeting was opened by Ender Muhammed Gümüş, Eskişehir Provincial Director of Agriculture and Forestry, the representative of the host institution. The project coordinator, Dr. Hatice Olcay, DVM, gave a presentation on the project activities, impacts and outcomes. During the week, the participants of the meeting had the opportunity to visit four dairy farms dedicated to the production of local cheese and other dairy products. They visited production facilities in Eskişehir (Yeşilçay Dairy Enterprise, TMR Food, Tarım Kredi Dairy Enterprise, Mahmudiye-Çifteler Milk Producers Association). In addition to the tours of the cheese production facilities, several lectures were given on technological processes and European hygiene standards in the production of traditional cheeses. Participants from the Ljubljana Veterinary Faculty gave lectures on the production methods of Tolminc, Mohant, Bovec and Nanos cheeses.

Further project meetings will take place in the European partner countries, the meeting is also planned in Slovenia. The conclusions of the project from this workshop will be the basis for the development of innovative and professional approaches that will lead to the modernization of milk processing and production in terms of quality and control standards in all EU countries. The main outcome of the project will be 10 lessons in five different languages, accessible to everyone, and two different training modules.

Primerjava mezenhimskih matičnih celic iz maščobnega tkiva psov in mačk

lukam — Mon, 25 Jan 2021 09:51:53 +0000

Comparison of Canine and Feline Adipose-Derived Mesenchymal Stem Cells/Medicinal Signaling Cells lukam Mon, 01/25/2021 - 10:55 Institute of Preclinical Sciences mesenchymal stem cells dog cat comparison proliferation differentiation cell surface marker viability Research

Comparison of Canine and Feline Adipose-Derived Mesenchymal Stem Cells/Medicinal Signaling Cells With Regard to Cell Surface Marker Expression, Viability, Proliferation, and Differentiation Potential.

Remarkable immunomodulatory abilities of mesenchymal stem cells, also called multipotent mesenchymal stromal cells or medicinal signaling cells (MSCs), have entailed significant advances in veterinary regenerative medicine in recent years. Despite positive outcomes from MSC therapies in various diseases in dogs and cats, differences in MSC characteristics between small animal veterinary patients are not well-known. We performed a comparative study of cells' surface marker expression, viability, proliferation, and differentiation capacity of adipose-derived MSCs (ADMSCs) from dogs and domestic cats. The same growth media and methods were used to isolate, characterize, and culture canine and feline ADMSCs. Adipose tissue was collected from 11 dogs and 8 cats of both sexes. The expression of surface markers CD44, CD90, and CD34 was detected by flow cytometry. Viability at passage 3 was measured with the hemocytometer and compared to the viability measured by flow cytometry after 1 day of handling. The proliferation potential of MSCs was measured by calculating cell doubling and cell doubling time from second to eighth passage. Differentiation potential was determined at early and late passages by inducing cells toward adipogenic, osteogenic, and chondrogenic differentiation using commercial media. Our study shows that the percentage of CD44+CD90+ and CD34−/− cells is higher in cells from dogs than in cells from cats. The viability of cells measured by two different methods at passage 3 differed between the species, and finally, canine ADMSCs possess greater proliferation and differentiation potential in comparison to the feline ADMSCs. The results show that in the development of new cell therapies it is necessary to take into account the animal species and the results of stem cell research cannot be easily transferred from one species to another.

Authors: Metka Voga, Valerija Kovač, Gregor Majdič

The full article is published in Frontiers in Veterinary Science

Nitrozativni stres v možganih psov s pasjo kognitivno motnjo

lukam — Mon, 18 Jan 2021 14:28:53 +0000

Nitrosative stress in brains from dogs with canine cognitive dysfunction lukam Mon, 01/18/2021 - 15:47 Institute of Preclinical Sciences Kognitivna motnja pes nitrozativni stres nNOS eNOS iNOS 3-nitrotirozin Research

Nitrosative stress in the frontal cortex from dogs with canine cognitive dysfunction.

Canine cognitive dysfunction (CCD) is an age-related disorder similar to human Alzheimer’s disease (AD) that occurs in elderly dogs. Nitrosative stress has been implicated as one of the causes leading to neurodegenerative diseases, particularly AD. Its involvement in the development of CCD has not been studied so far. In the present study, immunohistochemical staining detected all three isoforms of nitric oxide synthases (nNOS, eNOS, and iNOS) and 3-nitrotyrosine (3-NT) in brains from CCD-affected dogs and non-demented control dogs in all layers of the canine frontal cortex. In CCD-affected and non-demented brains, nNOS was highly expressed in pyramidal-like neurons in the upper cortical layers. nNOS has also been observed in astrocytes in the CCD frontal cortex. The nNOS immunohistochemical staining was statistically significantly elevated in dogs with CCD in comparison to non-demented dogs. Blood vessel wall cells were positive for eNOS, which was also expressed in astrocytes and neurons. Intense 3-NT immunoreactivity was observed in the upper cortical layers, where amyloid-beta deposits spread in the last stage of CCD. Brain cells in the same area were highly
immunoreactive for iNOS. This infers that neuroinflammation and nitrosative stress might exacerbate the neurodegenerative process in CCD-affected brains, ultimately leading to cognitive impairment.

Slika 1. nNOS protein v pasjih možganih. Slika prikazuje značilno obarvane živčne celice (rjavo) v čelnem režnju 16 letnega shih-tzuja s pasjo kognitivno motnjo. Merili sta dolgi 50 µm.

Slika 2. 3-NT v pasjih možganih. 3-NT (zeleno obarvanje) je prisoten v celicah, ki obkrožajo amiloide (Aβ) plake (rdeče obarvanje) pa tudi izven celic v območju plaka. Celična jedra so obarvana modro. Prikazan je čelni reženj 16 letnega jazbečarja s kognitivno motnjo. Merilo je dolgo 10 µm.

The full article is published in Frontiers in Veterinary Science

Authors: Sonja Prpar Mihevc, Maja Zakošek Pipan, Malan Štrbenc, Boris Rogelj, Gregor Majdič

Poliklorirani bifenili (PCB): prepovedani več kot 40 let, a zelo obstojni in še aktualni

lukam — Tue, 22 Dec 2020 08:55:52 +0000

Polychlorinated biphenyls (PCB): highly persistent chemicals banned for more than 40 years lukam Tue, 12/22/2020 - 10:06 Institute of Preclinical Sciences poliklorirani bifenili PCB trabekularna kostnina epifizni rastni hrustanec histomorfometrija podgana Research

Effects of PBC-169 and PCB-155 on microscopical structure of femur bone tissue of rat offspring.

Polychlorinated biphenyls (PCBs) are ubiquitous, persistent, organic pollutants also considered endocrine-disrupting chemicals. Our study examined the effects of lactational exposure to nondioxin-like PCB-155 and/or dioxin-like PCB-169 on longitudinal femur growth at the distal epiphyseal growth plate (EGP) in young rats at three different ages [postnatal days (PNDs) 9, 22, and 42]. After delivery, lactating rats were divided into four groups (PCB-169, PCB-155, PCB-155 + 169, and control) and administered PCBs intraperitoneally.

The femurs of offspring were used to estimate growth rate (µm/day), and histomorphometric analysis on the distal femur included the thickness of the EGP and zones of proliferation and hypertrophy with calcification. Stereometry was used to determine trabecular bone volume density. In the PCB-169 and PCB-155 + 169 groups, PCB-169 affected longitudinal bone growth in the early postnatal period by interfering with chondrocytes in the EGP zone of proliferation and, to a lesser extent, the zone of hypertrophy. Morphometric alterations in EGP structure diminished until puberty. A slow growth rate persisted in the PCB-169 group until PND 42, while in the PCB-155 group, a fast growth rate between PNDs 9 to 22 was significantly reduced between PNDs 22 to 42. Sterometric assessment showed decreased trabecular bone volume in the PCB-155 + 169 group compared with that in the control on PND 9 and increased in the PCB-169 group compared with that in the PCB-155 group on PND 42. To summarize, studied PCB congeners exerted congener- and age-dependent effects on femur growth rate and its histomorphometric characteristics.

The full article is published in Scientific Reports (Nature).

Authors: Jana Brankovič, Gregor Fazarinc, Janja Jan in Milka Vrecl

Kakšne poti endocitoze uporabljajo herpesvirusni, s proteinom G sklopljeni receptorji in kako jih ugotavljamo?

lukam — Mon, 14 Dec 2020 14:13:44 +0000

Which endocytotic pathways do herpesvirus encoded G-protein coupled receptors use and how do we study them? lukam Mon, 12/14/2020 - 15:23 Institute of Preclinical Sciences endocytosis G-protein coupled receptors herpesvirus Research

Methods for Studying Endocytotic Pathways of Herpesvirus Encoded G Protein-Coupled Receptors.

In mammalian cell, different physiological and pathological cellular processes are regulated by G protein-coupled receptors (GPCRs), a family of receptors expressed at the cell surface. Intriguingly, during evolution, several herpesviruses adopted genes for such receptors (viral GPCRs) from their hosts to benefit their own survival during infection. Viruses use these receptors to evade host immune system, to enter the cell and to transform infected cells and thereby establish widespread and lifelong infection; processes where receptor signaling and/or endocytosis seem central. Endocytosis is a fundamental process involved in trafficking of various extracellular and transmembrane molecules from the cell surface to its interior. This enables cells to communicate and respond to external environments, maintaining cellular homeostasis, and transduce signals. vGPCRs are rising as potential drug targets as exemplified by the cytomegalovirus-encoded receptor US28, where its constitutive internalization has been used for selective drug delivery in virus infected cells. This review provides an overview of the current knowledge of endocytic and cell localization properties of vGPCRs and methodological approaches used for studying receptor internalization. Using novel approaches, we show constitutive internalization of the BILF1 receptor from human and porcine γ-1 herpesviruses and we present motifs from the eukaryotic linear motif (ELM) resources with importance for vGPCR endocytosis. The fact that vGPCRs are only expressed at the surface of infected cells makes them attractive targets for antiviral therapy. With increasing knowledge on their trafficking and signaling properties, we are making step forward in discovering their druggable potential.

Authors: Maša Mavri, Katja Spiess, Mette Marie Rosenkilde, Catrin Sian Rutland, Milka Vrecl, Valentina Kubale

The full article is published in MDPI Molecules

Kakšno vlogo igrajo receptorji s sedmimi transmembranskimi območji pri zdravem in bolnem srcu?

lukam — Tue, 10 Nov 2020 06:08:16 +0000

Which role do seven transmembrane receptors have in healthy and broken heart? lukam Tue, 11/10/2020 - 07:15 Inštitut za predklinične vede angiotensin adrenorergic receptors cardiomyopathy heart disease endothelin-1 muscarinic receptors Research

The function of seven transmembrane receptors in the cardiovascular system and their role in the development of cardiomyopathy.

Avtor fotografije: Catrin Sian Rutland

The function of seven transmembrane receptors in the cardiovascular system and their role in the development of cardiomyopathy.

The G-protein-coupled receptors (GPCRs), also termed seven-transmembrane receptors (7TM-Rs) are a conserved family of transmembrane receptors, which are essential not only in the healthy heart and blood vessels but also for treatment of cardiovascular disease and failure. Heart failure is a leading global cause of morbidity and death and as such understanding 7TM-Rs, their functions, structures and potential for therapy essential. This review summarizes the roles of 7TM-Rs in the healthy functioning cardiovascular system and cardiac disorders with an emphasis in cardiomyopathy. It also discusses the significance of autoantibodies against the 7TM-Rs in cardiomyopathy.

Authors: Valentina Kubale, Ewelina Prozorowska, Kristýna Glocová, Lucy Slater, Catrin Sian Rutland

The full article is published in IntechOpen.