Applicability of MHC loci as markers for detecting hybridization between wolf (Canis lupus) and dog (Canis lupus familiaris)

Vukovi su česta pojava u blizini ljudskih naselja, koja naseljavaju i psi. Sve vrste roda Canis mogu stvarati plodno potomstvo, što znači da i vukovi i psi mogu stvarati hibride. Hibridizacija između divljih i udomaćenih vrsta, iako može imati i pozitivne posljedice, uglavnom se percipira kao prijetnja bioraznolikosti, najviše zbog introgresije alela domesticiranih vrsta, koji su nastali umjetnom selekcijom, u divlje populacije. Hibridi se ne mogu pouzdano identificirati samo po morfološkim obilježjima nego se koriste i molekularne metode. Uz uobičajenu genetičku analizu mitohondrijske DNA, Y kromosoma i mikrosatelita, 2012. godine prvi put je uspješno upotrijebljena analiza MHC lokusa kao metoda otkrivanja hibridizacije. U svrhu provjere primjenjivosti ove metode na hibride između vukova i pasa analizirani su aleli i haplotipovi DRB, DQA i DQB lokusa glavnog sustava tkivne podudarnosti 13 potencijalnih hibrida od kojih su tri bila ranije potvrđena kao hibridi nastali povratnim križanjem s vukom, i uspoređeni s podacima dobivenim za vuka i psa iz literature. U dva uzorka MHC lokusi nisu se pokazali informativnima za otkrivanje hibridizacije, dok su za jedan uzorak bili informativni. MHC lokusi mogu biti dobri biljezi za otkrivanje hibridizacije vuka i psa jedino u slučaju kad potecijalni hibridi nose privatne alele za pojedinu vrstu.Wolves can often be seen near human settlements, where dogs live. All Canis species are closely related and can produce fertile progeny. Hybridization between wild and domesticated species can have a positive outcome, but it is usually percieved as threat to biodiversity, mostily because of the possibility of introgression of artificially selected domestic alleles into wild populations. Hybrids can't be identified solely based on morphological features, so using molecular methods is necessary. In 2012, along with the usual analysis of the mithochondrial DNA, Y chromosome and microsatellites, MHC loci analysis was used to detect hybrids for the first time. In order to find out if this method is applicable in detecting wolf-dog hybrids, DRB, DQA, DQB alleles and haplotypes of 13 suspected hybrids, three of which were earlier confirmed as back – crosses with wolves, were analysed and compared to available data from existing literature. MHC loci were informative in detecting hybridization for one of those samples, but weren't for other two. MHC loci are effective markers only when potential hybrids are carrying private alleles for each species

Why humans have no fur?

Nakon što se evolucijska linija čovjeka odvojila od predaka čimpanzi, započela je preobrazba u „golog majmuna”. Čovjek je izgubio krzno zbog promjene klime. Zahvaljujući globalnom zahladnjenju, klima je postala suša, čovjek je izašao u sunčanu savanu i počeo živjeti aktivnijim životom. Pod prijetnjom pregrijavanja izgubio je krzno i povećao broj ekrinih žlijezda u koži. To mu je omogućilo da znojenjem efikasno ohladi tijelo. Ipak, na pojedinim dijelovima tijela ostale su dlake, s razlikama između populacija i muškaraca i žena. Ovaj rad razmatra kako, kada i zašto je čovjek ostao bez krzna, koje su posljedice i koje velike evolucijske promjene je ovaj događaj omogućio. Većinski golo tijelo bilo je okidač za ključne korake u daljnjoj evoluciji čovjeka. Omogućilo je jasno odvajanje čovjeka od ostalih životinja.After human evolutionary lineage parted ways with that of chimpanzee's ancestors, transformation into a naked ape began. Humans lost their fur due to a climate change. Thanks to the global cooling, climate became more arid, people came out to sunny savannas and started living a more active lifestyle. Under threat of overheating, they lost their fur and increased the number of ecrine glands in their skin. That enabled efficiant cooling down of the body by sweating. Still, some parts of the human body kept their hair, with some differences between populations and between males and females. This paper considers how, when and why humans lost their fur, what are the consequences and which great changes could now happen. Mostly naked body triggered key events in future human evolution. It made clear separation of people and other animals possible

Trefoil Factor 3 Protein and Sepsis – A Review

Sepsis is one of the most common causes of death in hospitalized patients. Disruption of intestinal barrier homeostasis is one of its main hallmarks. Trefoil factor family proteins are known for their role in protecting and repairing the intestinal mucosa. It has been repeatedly shown that the TFF3 protein is involved in maintaining the intestinal barrier. For that reason, it has been studied in the search for objective measures to predict the onset or outcome of sepsis. Several studies have been performed on rodent sepsis models and on sepsis patients, both children and adults. From the limited research available to date, it appears that TFF3 is involved in the pathogenesis of sepsis, but the exact mechanism is not yet clear. Its potential as a sepsis biomarker has so far been low, but more extensive studies on its role in predicting disease severity and outcome, as well as organ dysfunction, may lead to finding specific patient groups or sepsis stages for which it would be suitable

Tff3 Deficiency Protects against Hepatic Fat Accumulation after Prolonged High-Fat Diet

Trefoil factor 3 (Tff3) protein is a small secretory protein expressed on various mucosal surfaces and is involved in proper mucosal function and recovery via various mechanisms, including immune response. However, Tff3 is also found in the bloodstream and in various other tissues, including the liver. Its complete attenuation was observed as the most prominent event in the early phase of diabetes in the polygenic Tally Ho mouse model of diabesity. Since then, its role in metabolic processes has emerged. To elucidate the complex role of Tff3, we used a new Tff3-deficient mouse model without additional metabolically relevant mutations (Tff3-/-/C57BL/6NCrl) and exposed it to a high-fat diet (HFD) for a prolonged period (8 months). The effect was observed in male and female mice compared to wild-type (WT) counter groups (n = 10 animals per group). We monitored the animals’ general metabolic parameters, liver morphology, ultrastructure and molecular genes in relevant lipid and inflammatory pathways. Tff3-deficient male mice had reduced body weight and better glucose utilization after 17 weeks of HFD, but longer HFD exposure (32 weeks) resulted in no such change. We found a strong reduction in lipid accumulation in male Tff3-/-/C57BL/6NCrl mice and a less prominent reduction in female mice. This was associated with downregulated peroxisome proliferator-activated receptor gamma (Pparγ) and upregulated interleukin-6 (Il-6) gene expression, although protein level difference did not reach statistical significance due to higher individual variations. Tff3-/-/C57Bl6N mice of both sex had reduced liver steatosis, without major fatty acid content perturbations. Our research shows that Tff3 protein is clearly involved in complex metabolic pathways. Tff3 deficiency in C57Bl6N genetic background caused reduced lipid accumulation in the liver ; further research is needed to elucidate its precise role in metabolism-related events

Effect of Tff3 Deficiency and ER Stress in the Liver

Endoplasmic reticulum (ER) stress, a cellular condition caused by the accumulation of unfolded proteins inside the ER, has been recognized as a major pathological mechanism in a variety of conditions, including cancer, metabolic and neurodegenerative diseases. Trefoil factor family (TFFs) peptides are present in different epithelial organs, blood supply, neural tissues, as well as in the liver, and their deficiency has been linked to the ER function. Complete ablation of Tff3 expression is observed in steatosis, and as the most prominent change in the early phase of diabetes in multigenic mouse models of diabesity. To elucidate the role of Tff3 deficiency on different pathologically relevant pathways, we have developed a new congenic mouse model Tff3-/-/C57BL6/N from a mixed background strain (C57BL6/N /SV129) by using a speed congenics approach. Acute ER stress was evoked by tunicamycin treatment, and mice were sacrificed after 24 h. Afterwards the effect of Tff3 deficiency was evaluated with regard to the expression of relevant oxidative and ER stress genes, relevant proinflammatory cytokines/chemokines, and the global protein content. The most dramatic change was noticed at the level of inflammation-related genes, while markers for unfolded protein response were not significantly affected. Ultrastructural analysis confirmed that the size of lipid vacuoles was affected as well. Since the liver acts as an important metabolic and immunological organ, the influence of Tff3 deficiency and physiological function possibly reflects on the whole organism

Impact of High Salt Diet on Cerebral Vascular Function and Stroke in Tff3−/−/C57BL/6N Knockout and WT (C57BL/6N) Control Mice

High salt (HS) dietary intake leads to impaired vascular endothelium-dependent responses to various physiological stimuli, some of which are mediated by arachidonic acid (AA) metabolites. Transgenic Tff3−/− gene knockout mice (Tff3−/−/C57BL/6N) have changes in lipid metabolism which may affect vascular function and outcomes of stroke. We aimed to study the effects of one week of HS diet (4% NaCl) on vascular function and stroke induced by transient occlusion of middle cerebral artery in Tff3−/− and wild type (WT/C57BL/6N) mice. Flow-induced dilation (FID) of carotid artery was reduced in WT-HS mice, but not affected in Tff3−/−-HS mice. Nitric oxide (NO) mediated FID. NO production was decreased with HS diet. On the contrary, acetylcholine-induced dilation was significantly decreased in Tff3−/− mice on both diets and WT-HS mice. HS intake and Tff3 gene depletion affected the structural components of the vessels. Proteomic analysis revealed a significant effect of Tff3 gene deficiency on HS diet-induced changes in neuronal structural proteins and acute innate immune response proteins’ expression and Tff3 depletion, but HS diet did not increase the stroke volume, which is related to proteome modification and upregulation of genes involved mainly in cellular antioxidative defense. In conclusion, Tff3 depletion seems to partially impair vascular function and worsen the outcomes of stroke, which is moderately affected by HS diet

Applicability of MHC loci as markers for detecting hybridization between wolf (Canis lupus) and dog (Canis lupus familiaris)

Vukovi su česta pojava u blizini ljudskih naselja, koja naseljavaju i psi. Sve vrste roda Canis mogu stvarati plodno potomstvo, što znači da i vukovi i psi mogu stvarati hibride. Hibridizacija između divljih i udomaćenih vrsta, iako može imati i pozitivne posljedice, uglavnom se percipira kao prijetnja bioraznolikosti, najviše zbog introgresije alela domesticiranih vrsta, koji su nastali umjetnom selekcijom, u divlje populacije. Hibridi se ne mogu pouzdano identificirati samo po morfološkim obilježjima nego se koriste i molekularne metode. Uz uobičajenu genetičku analizu mitohondrijske DNA, Y kromosoma i mikrosatelita, 2012. godine prvi put je uspješno upotrijebljena analiza MHC lokusa kao metoda otkrivanja hibridizacije. U svrhu provjere primjenjivosti ove metode na hibride između vukova i pasa analizirani su aleli i haplotipovi DRB, DQA i DQB lokusa glavnog sustava tkivne podudarnosti 13 potencijalnih hibrida od kojih su tri bila ranije potvrđena kao hibridi nastali povratnim križanjem s vukom, i uspoređeni s podacima dobivenim za vuka i psa iz literature. U dva uzorka MHC lokusi nisu se pokazali informativnima za otkrivanje hibridizacije, dok su za jedan uzorak bili informativni. MHC lokusi mogu biti dobri biljezi za otkrivanje hibridizacije vuka i psa jedino u slučaju kad potecijalni hibridi nose privatne alele za pojedinu vrstu.Wolves can often be seen near human settlements, where dogs live. All Canis species are closely related and can produce fertile progeny. Hybridization between wild and domesticated species can have a positive outcome, but it is usually percieved as threat to biodiversity, mostily because of the possibility of introgression of artificially selected domestic alleles into wild populations. Hybrids can't be identified solely based on morphological features, so using molecular methods is necessary. In 2012, along with the usual analysis of the mithochondrial DNA, Y chromosome and microsatellites, MHC loci analysis was used to detect hybrids for the first time. In order to find out if this method is applicable in detecting wolf-dog hybrids, DRB, DQA, DQB alleles and haplotypes of 13 suspected hybrids, three of which were earlier confirmed as back – crosses with wolves, were analysed and compared to available data from existing literature. MHC loci were informative in detecting hybridization for one of those samples, but weren't for other two. MHC loci are effective markers only when potential hybrids are carrying private alleles for each species

Tff3 deficiency differentially affects the morphology of male and female intestines in a long-term high-fat-diet-fed mouse model

Trefoil factor family protein 3 (Tff3) is a small peptide (59 amino acids7 kDa) that is a member of the trefoil factor family proteins (Tffs), comprising the Tff1, Tff2, and Tff3 proteins [1]. Tffs share a conserved motif of a well-defined three-loop structure, reminiscent of a trefoil stabilized by three disulfide bonds. Tffs are predominantly expressed in the gastrointestinal tract, where they protect the mucosal surface [2]. The presence of Tff3 in the bloodstream and various other organs, including the liver, brain, pancreas, and lymphoid tissue, indicates its general importance in organisms [3,4]. The mode of action of Tffs ranges from a simple increase in mucous viscosity, cytoprotection, and antiapoptotic and chemotactic effects to more complex immune regulatory functions [5,6]. Tff expression patterns are altered in various tumors, implying a strong association with tumorigenesis [7]. Tffs were long thought to be involved in mucosal repair by promoting cell migration (“restitution”) via their weak chemotactic and anti-apoptotic effects. Recent data on the molecular forms of human Tffs have revealed a more complex situation. Tff1 and Tff3 occur in vivo in different molecular forms and can form disulfide-linked heterodimers. In the intestine, Tff3 occurs as a Tff3-FCGBP (Fc fragment of the IgG binding protein (FCGBP)) heterodimer, Tff3-Tff3 homodimer, and Tff3 monomer [8,9,10,11]. This indicates that the biological functions of Tff peptides are complex [12]. Additionally, minimal amounts of Tff peptides are secreted in an endocrine manner, for example, by the central nervous system (CNS) [12] and immune system [13,14]. The potential role of the Tff3 gene in a metabolism-related condition called diabesity (a combination of diabetes and obesity) was first revealed by quantitative trait locus (QTL) analysis of a mouse model of diabetes, the Tally Ho mouse strain [14], in which the complete reduction of Tff3 in the liver was the most dramatic change in early diabetes. Regulation of Tff3 with appropriate food intake and improvement in glucose tolerance in a diet-induced obesity model raises additional questions regarding the involvement of Tff3 in metabolic pathways [15]. This raises the question of whether Tff3 participates in the gut–liver–brain (GLB) axis. Animal mouse models of Tff protein deficiency are crucial for elucidating the physiological functions of specific proteins. Tff3-deficient mice show markedly increased sensitivity in a dextran sulfate sodium (DSS) colitis model, and these animals are particularly sensitive to radiation-induced mucosal injury and chemotherapy [16,17]. Tff3-deficient mice with a mixed genetic background (sv129/C57BL/6J) exhibit altered liver lipid metabolism [18]. It is well known that the phenotype of a given single gene mutation in mice is modulated by the genetic background of the inbred strain in which the mutation is maintained. This effect is attributable to the modifier genes, which act in combination with the causative gene [19]. A previously existing Tff3-deficient mouse strain had a mixed [18] or C57BL/6J background [16]. C57BL/6J mice have additional genetic variations in the nicotinamide nucleotide transhydrogenase (Nnt) gene, which encodes a mitochondrial redox-induced proton pump that links NADPH synthesis to the mitochondrial metabolic pathway. The Nnt mutation itself modulates metabolism [20,21] and the immune response [22] and hides the physiological function of the investigated candidate protein. The mixed genetic background poses the problem of ensuring a proper wild-type (Wt) control group due to random combinations of genetic variations [23]. Using the Tff3-/-//C57BL/6N mouse model, we overcame the issues of mixed genetic backgrounds and additional mutations in the C57BL/6J strain. Since the Tff3 protein is present in the blood circulation, intestine, liver, and brain, its role in these organs and the pathology of the GLB axis is of great interest. This study aimed to determine the effect of Tff3 deficiency on the intestines of male and female mice under metabolically relevant conditions of long-term consumption of a high-fat diet (HFD) known to have pathological effects on the GLB axis. To exclude the effects of additional mutations and mixed genetic variations, we used a new congenic Tff3-/- strain on the C57BL/6NCrl genetic background with no additional metabolism-relevant mutations [24]. As existing data link Tff3 to metabolism, it is of great interest to investigate the systemic role of Tff3 deficiency in long-term HFD feeding (36 weeks). In this study, we monitored the effects of long-term HFD feeding on intestinal morphology and microbiome-relevant short-chain fatty acid (SCFA) content. The effect of Tff3 deficiency on the expression of genes involved in Tff3 function-related pathways (oxidative and endoplasmic reticulum (ER) stress, apoptosis, and inflammation) was determined using quantitative PCR (qPCR). Since Tff3 is regulated by estrogens [25] and metabolic events are influenced by sex, we monitored the impact of long-term HFD consumption on Wt and Tff3-deficient male and female mice