Impairment of the Hif-1α regulatory pathway in Foxn1-deficient (Foxn1-/- ) mice affects the skin wound healing process.

Hypoxia and hypoxia-regulated factors (eg, hypoxia-inducible factor-1α [Hif-1α], factor inhibiting Hif-1α [Fih-1], thioredoxin-1 [Trx-1], aryl hydrocarbon receptor nuclear translocator 2 [Arnt-2]) have essential roles in skin wound healing. Using Foxn1-/- mice that can heal skin injuries in a unique scarless manner, we investigated the interaction between Foxn1 and hypoxia-regulated factors. The Foxn1-/- mice displayed impairments in the regulation of Hif-1α, Trx-1, and Fih-1 but not Arnt-2 during the healing process. An analysis of wounded skin showed that the skin of the Foxn1-/- mice healed in a scarless manner, displaying rapid re-epithelialization and an increase in transforming growth factor ß (Tgfß-3) and collagen III expression. An in vitro analysis revealed that Foxn1 overexpression in keratinocytes isolated from the skin of the Foxn1-/- mice led to reduced Hif-1α expression in normoxic but not hypoxic cultures and inhibited Fih-1 expression exclusively under hypoxic conditions. These data indicate that in the skin, Foxn1 affects hypoxia-regulated factors that control the wound healing process and suggest that under normoxic conditions, Foxn1 is a limiting factor for Hif-1α.

In Silico Identification of lncRNAs Regulating Sperm Motility in the Turkey (Meleagris gallopavo L.).

Long non-coding RNAs (lncRNAs) are transcripts not translated into proteins with a length of more than 200 bp. LncRNAs are considered an important factor in the regulation of countless biological processes, mainly through the regulation of gene expression and interactions with proteins. However, the detailed mechanism of interaction as well as functions of lncRNAs are still unclear and therefore constitute a serious research challenge. In this study, for the first time, potential mechanisms of lncRNA regulation of processes related to sperm motility in turkey were investigated and described. Customized bioinformatics analysis was used to detect and identify lncRNAs, and their correlations with differentially expressed genes and proteins were also investigated. Results revealed the expression of 863 new/unknown lncRNAs in ductus deferens, testes and epididymis of turkeys. Moreover, potential relationships of the lncRNAs with the coding mRNAs and their products were identified in turkey reproductive tissues. The results obtained from the OMICS study may be useful in describing and characterizing the way that lncRNAs regulate genes and proteins as well as signaling pathways related to sperm motility.

Characterization and biological role of cysteine-rich venom protein belonging to CRISPs from turkey seminal plasma†.

Turkey semen contains cysteine-rich secretory proteins (CRISPs) that belong to the dominant seminal plasma proteins. We aimed to isolate and characterize CRISP from turkey seminal plasma and evaluate its possible involvement in yellow semen syndrome (YSS). YSS, which is well characterized, causes reduced fertility and hatchability. The protein was purified using hydrophobic interaction, gel filtration, and reverse phase chromatography. It then was subjected to identification by mass spectrometry, analysis of physicochemical properties, and specific antibody production. The biological function of the isolated protein was tested and included its effects on sperm motility and migration and sperm-egg interactions. Sperm motility was measured with the CASA system using Hobson Sperm Tracker. The reproductive tract of turkey toms was analyzed for gene expression; immunohistochemistry was used for protein localization in the male reproductive tract, spermatozoa, and inner perivitelline layer. The isolated protein was identified as cysteine-rich venom protein-like isoform X2 (CRVP X2; XP_010706464.1) and contained feature motifs of CRISP family proteins. Turkey CRVP X2 was present in both spermatozoa and seminal plasma. The extensive secretion of CRVP X2 by the epithelial cells of the epididymis and ductus deferens suggests its involvement in post-testicular sperm maturation. The internally localized CRVP X2 in the proximal part of the sperm tail might be responsible for stimulation of sperm motility. CRVP X2 on the sperm head might be involved in several events prior to fusion and may also participate in gamete fusion itself. Although the mechanisms by which CRVP X2 mediates fertilization are still unknown, the involvement of complementary sites cannot be excluded. The disturbance of CRVP X2 expression can serve as an etiologic factor of YSS in the turkey. This study expands the understanding of the detailed mechanism of fertilization in birds by clarifying the specific role of CRVP X2.

Wnt signaling and the transcription factor Foxn1 contribute to cutaneous wound repair in mice.

Aim: The transcription factor Foxn1 is a regulator of scar-ended cutaneous wound healing in mice. However, the link between Foxn1 and Wnt signaling has not been explored in the context of cutaneous repair. Here, we investigate the effects of ß-catenin-dependent and -independent Wnt signaling represented by Wnt10a and Wnt11, respectively, in healing of full-thickness cutaneous wounds in C57BL/6 mice. Material and Methods: Quantitative polymerase chain reaction, western blot, and immunostaining were performed to assess the spatial and temporal distribution of Wnt10a, Wnt11, and ß-catenin in skin during wound healing. A co-culture system consisting of keratinocytes transfected with an adenoviral vector carrying Foxn1-GFP and dermal fibroblasts (DFs) was employed to determine the influence of epidermal signals on the capacity of DFs to produce extracellular matrix (ECM) proteins in vitro. The levels of types I and III collagen in conditioned media from DFs cultures were examined via enzyme-linked immunosorbent assay. Results: The expression of Wnt10a, Wnt11, and ß-catenin increased at post-wounding days 14 and 21 when tissue remodeling occurred. Foxn1::Egfp transgenic mice experiments demonstrated that Wnts were abundant in the epidermis adjacent to the wound margin and to a lesser extent in the dermis. The Wnt10a signal colocalized with Foxn1-eGFP in the epithelial tongue and neo-epidermis during the initial stage of wound healing. Foxn1 overexpression in keratinocytes affected DFs function related to collagen synthesis. Conclusions: Wnt ligands contribute to cutaneous wound repair, predominantly by engagement in ECM maturation. The data indicates a possible relationship between Foxn1 and Wnts in post-traumatic skin tissue.

Age, Diet and Epidermal Signaling Modulate Dermal Fibroblasts' Adipogenic Potential.

The recognition of a distinct fat depot, the dermal white adipose tissue (dWAT), points out the complexity of the interaction among skin resident cells: keratinocytes, dermal fibroblasts (DFs) and adipocytes in response to physiological (diet, age) and pathological (injury) stimulations. dWAT has been recognized as a significant contributor to thermoregulation, hair cycle, immune response, wound healing and scarring. In this study, we examined age- and diet-related changes in dWAT modulation and DFs' adipogenic potential. The data showed that diet modulates dWAT expansion predominantly by hypertrophy, whereas age affects the pool of adipocyte progenitor cells in the skin indicating its role in dWAT hyperplasia. Analysis of DFs' migratory abilities in the model of skin explants isolated from the skin of young, old, low (LFD)- or high (HFD)-fat diet C56BL/6 mice revealed that HFD, regardless of animal age has the most profound stimulatory impact of DF migration. We determined that the adipogenic potential of DFs is comparable to stromal vascular fraction (SVF) of inguinal fat depot and ear mesenchymal stem cells (EMSC). We also showed the stimulatory role of epidermally expressed transcription factor Foxn1 on adipogenic signaling: bone morphogenetic protein 2 (Bmp2) and insulin-like growth factor 2 (Igf2) in keratinocytes.

Effect of TGFß1, TGFß3 and keratinocyte conditioned media on functional characteristics of dermal fibroblasts derived from reparative (Balb/c) and regenerative (Foxn1 deficient; nude) mouse models.

Skin injuries in mammals are healed through repair or regeneration. Our previous studies demonstrated that deficient expression of the transcription factor Foxn1 in epidermis of nude mice accounts for their skin's pronounced regenerative properties. Since homeostasis within the skin depends on complex interactions between the epidermal and underlying dermal layers, the present study characterizes and compares isolated dermal fibroblasts (DFs) between regenerative nude (Foxn1 deficient) mice and their wild-type Balb/c counterparts. Nude DFs exhibited a higher cumulative number of population doublings (cumulative PD) at low seeding density and increased adipogenic differentiation capacity relative to their Balb/c DF counterparts. Nude DFs displayed reduced migration and gel contraction, functional features associated with wound healing. The comparison of transforming growth factor ß family (TGFß) expression showed significantly higher levels of Tgfß3 transcript between nude and Balb/c mice but no differences were detected for Tgfß1. Nude DFs were specifically sensitive to the presence of the pro-regenerative TGFß3 isoform, showing increased collagen I deposition and alpha smooth muscle actin expression. Viability of Balb/c DFs was stimulated by keratinocyte conditioned media (KCM) from Balb/c (Foxn1 active) but inhibited by nude (Foxn1 deficient) KCM. In contrast, nude DFs did not respond to either KCMs with respect to their metabolic activity. Collectively, the enhanced plasticity and greater sensitivity of nude DFs to TGFß3 stimulation are indicative of and consistent with their pro-regenerative characteristics. These data support the hypothesis that epidermal Foxn1 plays a critical role in determining the DFs regenerative phenotype.

Bone Marrow Adipocyte Developmental Origin and Biology.

PURPOSE OF REVIEW: This review explores how the relationships between bone marrow adipose tissue (BMAT) adipogenesis with advancing age, obesity, and/or bone diseases (osteopenia or osteoporosis) contribute to mechanisms underlying musculoskeletal pathophysiology. RECENT FINDINGS: Recent studies have re-defined adipose tissue as a dynamic, vital organ with functions extending beyond its historic identity restricted solely to that of an energy reservoir or sink. "State of the art" methodologies provide novel insights into the developmental origin, physiology, and function of different adipose tissue depots. These include genetic tracking of adipose progenitors, viral vectors application, and sophisticated non-invasive imaging modalities. While constricted within the rigid bone cavity, BMAT vigorously contributes to local and systemic metabolic processes including hematopoiesis, osteogenesis, and energy metabolism and undergoes dynamic changes as a function of age, diet, bone topography, or sex. These insights will impact future research and therapies relating to osteoporosis.

Foxn1 in Skin Development, Homeostasis and Wound Healing.

Intensive research effort has focused on cellular and molecular mechanisms that regulate skin biology, including the phenomenon of scar-free skin healing during foetal life. Transcription factors are the key molecules that tune gene expression and either promote or suppress gene transcription. The epidermis is the source of transcription factors that regulate many functions of epidermal cells such as proliferation, differentiation, apoptosis, and migration. Furthermore, the activation of epidermal transcription factors also causes changes in the dermal compartment of the skin. This review focuses on the transcription factor Foxn1 and its role in skin biology. The regulatory function of Foxn1 in the skin relates to physiological (development and homeostasis) and pathological (skin wound healing) conditions. In particular, the pivotal role of Foxn1 in skin development and the acquisition of the adult skin phenotype, which coincides with losing the ability of scar-free healing, is discussed. Thus, genetic manipulations with Foxn1 expression, specifically those introducing conditional Foxn1 silencing in a Foxn1+/+ organism or its knock-in in a Foxn1−/− model, may provide future perspectives for regenerative medicine.

Foxn1 and Mmp-9 expression in intact skin and during excisional wound repair in young, adult, and old C57Bl/6 mice.

The transcription factor Foxn1 is essential for skin development. Our previous studies performed on young C57BL/6J mice model showed that Foxn1 acts as regulator of the skin wound healing process. The present study extended our initial research regarding the expression and potential role of Foxn1 in the intact and wounded skin as a function of animal age and stage of the wound healing process. We analyzed Foxn1 and Mmp-9 expression in the intact and postinjured skin of young, adult, and old C57BL/6J and transgenic Foxn1::Egfp mice. The similar levels of epidermal Foxn1 mRNA expression were detected in young and adult C57BL/6J mice and higher levels in old animals. Postinjured skin tissues displayed a gradual decrease of Foxn1 mRNA expression at Days 1, 5, and 7 after injury. Foxn1-eGFP positive cells were abundant at wound margin and in re-epithelialized epidermis at postwounded Days 1, 5, and 7 and colocalized with E-cadherin and Mmp-9. Postwounded skin at Days 14-36 displayed Foxn1-eGFP cells in the epidermis and in the dermal part of the skin (papillary dermis). A subset of Foxn1-eGFP positive cells in the papillary dermis expressed the myofibroblast marker αSMA. Flow cytometric analysis of cells isolated from postwounded (Day 7) skin tissues showed a significant increase in the percentage of Foxn1-eGFP positive cells with phenotype of double positivity for E-cadherin/N-cadherin (epithelial/mesenchymal markers). Collectively, these data identify the transcription factor Foxn1 as a potential key epidermal regulator modifying both epidermal and dermal healing processes after cutaneous wounding.

Evidence for the presence of stem/progenitor cells in porcine endometrium.

A population of adult stem cells responsible for cyclic reconstructing and remodeling has been proposed to reside in the highly regenerative mammalian endometrium. Recently, stem/progenitor cells have been identified in the human and mouse endometrium, but less is known about these cells in livestock animals. Using Hoechst 33342 fluorescent dye staining and flow cytometry, we identified an emerging cell side population that may be responsible for the regeneration process of the porcine endometrium. The percentage of side-population cells on Day 19 of the estrous cycle was significantly higher than that on Days 2-4. Moreover, single cells were able to seed clones that could differentiate into three independent mesenchymal-cell lineages. We also demonstrated the expression of specific markers of self-renewal cells on these side-population cells and the presence of a population of cells among the stromal cells that possess markers for mesenchymal stem cells. These results indicate that the porcine endometrium contains a population of cells with the capacity for self-renewal and a high rate of proliferation, which depend on the phase of the estrous cycle. These cells could potentially be involved in the cyclic reconstruction of the porcine endometrium.

Isolation and characterization of an ovoinhibitor, a multidomain Kazal-like inhibitor from Turkey (Meleagris gallopavo) seminal plasma.

Turkey seminal plasma contains three serine proteinase inhibitors. Two of them, with low molecular masses (6 kDa), were identified as single-domain Kazal-type inhibitors responsible for regulating acrosin activity. Our experimental objective was to isolate and characterize the inhibitor with the high molecular weight from turkey seminal plasma. The inhibitor was purified using hydrophobic interaction and affinity chromatography. Pure preparations of the inhibitor were used for identification by mass spectrometry, for determination of physicochemical properties (molecular weight, pI, and content and composition of the carbohydrate component), for kinetic studies, and for antibacterial tests. Gene expression and immunohistochemical detection of the inhibitor were analyzed in the testis, epididymis, and ductus deferens. The inhibitor with a high molecular weight from turkey seminal plasma was identified as an ovoinhibitor, which was found in avian semen for the first time. The turkey seminal plasma ovoinhibitor was a six-tandem homologous Kazal-type domain serine proteinase inhibitor that targeted multiple proteases, including subtilisin, trypsin, and elastase, but not acrosin. Our results suggested that hepatocyte growth factor activator was a potential target proteinase for the ovoinhibitor in turkey seminal plasma. The presence of the ovoinhibitor within the turkey reproductive tract suggested that its role was to maintain a microenvironment for sperm in the epididymis and ductus deferens. The turkey seminal plasma ovoinhibitor appeared to play a significant role in an antibacterial semen defense against Bacillus subtilis and Staphylococcus aureus.

Dental Malocclusion in Mixed Dentition Children and Its Relation to Podal System and Gait Parameters.

BACKGROUND: Dental malocclusion is an increasingly frequent stomatognathic disorder in children and adolescents nowadays. The purpose of this study was to confirm or deny the correlations between body posture and malocclusion. METHODS: In the study, gait, distribution of foot pressure on the ground, and body balance were examined. The research group consisted of 76 patients aged 12-15 years. The research group was obtained from patients attending periodic dental check-ups at Healthcare Center Your Health EL who agreed to participate in the study. The patients were divided into two groups without malocclusion and with malocclusion, using Angle classification, which enabled determination of the anteroposterior relationship of the first molars. The pedobarographic mat was used to analyze the distribution of foot forces on the ground, the diagnostic system Wiva® Science was used for gait analysis, and Kineod 3D was used for posture analysis. The Shapiro-Wilk test used for analysis showed inconsistency with normal distribution for all measurement parameters. The Mann-Whitney U test was used for the analysis, and the significance level was set at p ≤ 0.05. RESULTS: Examination of the relationship between stabilometric and gait parameters showed that the position of the mandible in relation to maxilla has an important effect on gait rhythm, gait cycle duration, and right step duration time. Patients diagnosed with malocclusion showed high-speed walking rhythm in comparison to patients with Angle's class I (p = 0.010). The duration of the whole gait cycle (p = 0.007) and the duration of right step (p = 0.027) were prolonged in students without orthodontic disorders compared to the other. CONCLUSIONS: The conducted study proved that there is correlation between the presence of a stomatognathic disorder and gait cycle parameters. There is a statistically noticeable correspondence between the position of the mandible in relation to maxilla and walking rhythm, gait cycle duration, and right step duration time. Namely, students who presented malocclusion had a high-speed walking rhythm and decreased duration of the gait cycle and of the right step. On the other hand, students without disorders (Angle's class I) showed low-speed rhythm and increased duration of the gait cycle and of the right step in comparison to Angle's classes II and III.

Changes in Gait Parameters and the Podal System Depending on the Presence of a Specific Malocclusion Type in School-Age Children.

BACKGROUND: The correspondence between various aspects of human postural parameters and the spatial relation of the jaws is of increasing interest among scientists. Emerging research suggests that the stomatognathic system and posture play, in a broad sense, significant roles. OBJECTIVES: The aim of the study was to analyze the relationship between various malocclusion types and gait parameters, the distribution of foot pressure on the ground, and body balance. METHODS: The study involved 155 patients aged 12-16. The subjects were divided into groups according to their malocclusion-Angle's class II (n = 32), Canine class II (n = 31), and Overbite (n = 46). The control group (n = 46) comprised children not demonstrating any defects. The study data were collected by direct observation of the oral cavity. Gait analysis was carried out using the Wiva® Science sensor, and the distribution of foot forces on the ground and body balance was determined via the E.P.S R/1 pedobarographic mat. The non-parametric Mann-Whitney U test was applied for statistical analysis. RESULTS: Analysis of the results obtained showed statistically significant differences in left step duration (p = 0.042) and the duration of the right step (p = 0.021), as well as the projection of the body's center of gravity on the left foot (p = 0.027). CONCLUSIONS: Distoocclusion in the anterior part of the mandible may cause different positioning of the head and neck, as well as varying tension of the muscles, further leading to balance disorders while walking.

Plantar pressure distribution and postural balance in beauty pageant contestants before and after wearing high-heeled shoes.

PURPOSE: The aim of this study was to assess the biomechanics of the foot in beauty pageant contestants wearing shoes that increase calcaneal height. Plantar pressure distribution and postural balance were examined in women after one hour of wearing high-heeled shoes. METHODS: Foot arch measurements were conducted using EPSR1. Based on these measurements, 18 of the 19 tested women were diagnosed with significant pes cavus. RESULTS: The experiment revealed a significant reduction in the excessively high metatarsal arch of both feet. The mean load on the metatarsus measured at rest was 0.0% in both feet, and it increased by 0.6% in the left leg (p ≤ 0.022; Z = 2.293) and by 2.7% in the right leg (p ≤ 0.023; Z = 2.271). These results suggest that excessive stretching of the plantar fascia and impaired function of the short muscles of the foot lead to a temporary reduction in the arch of the foot. This phenomenon can be compared to stretching a bowstring and its return to the shortest length under static conditions. A statistical analysis of the results of stabilographic measurements revealed significant changes only in the left foot barycenter. An analysis of the ability to maintain postural balance revealed that the observed changes contributed to postural destabilization at p ≤ 0.011 and Z = 2.535. CONCLUSIONS: Further research involving electromyography tests is needed to examine plantar muscle tension during isotonic contractions. The duration of the exercise-induced reduction in pes cavus should also be determined for therapeutic purposes.

Characterization of bovine immortalized luteal endothelial cells: action of cytokines on production and content of arachidonic acid metabolites.

BACKGROUND: The interactions between luteal, vascular endothelial, immune cells and its products: steroids, peptide hormones, prostaglandins (PGs), growth factors and cytokines play a pivotal role in the regulation of corpus luteum (CL) function. Luteal endothelial cells undergo many dynamic morphological changes and their action is regulated by cytokines. The aims are: (1) to establish in vitro model for bovine luteal endothelial cells examination; (2) to study the effect of cytokines: tumor necrosis factor alpha (TNFalpha) and interferon gamma (IFNgamma) on cell viability, leukotrienes (LTs) and PG synthases, and endothelin-1 (EDN-1) mRNA, protein expression and their secretion in bovine immortalized luteal endothelial (EnCL-1) cells. METHODS: The primary cultures of bovine luteal endothelial cells were immortalized by transfection with vector carrying the Simian virus 40 T-antigen (SV40 T-ag) sequence. Expression of SV40 T-ag gene in EnCL-1 cells was confirmed by RT-PCR and immunofluorescence staining showed the presence of endothelial cell markers: VE-cadherin and von Willebrand factor. EnCL-1 cells were stimulated by TNFalpha with IFNgamma (50 ng/ml each) for 24 h. Cell viability, mRNA expression (real time RT-PCR), protein expression (western blotting) for LTC4 synthase (LTC4S), LTA4 hydrolase (LTA4H), PGE2 and PGF2alpha synthases and endothelin-1 (EDN-1), and levels of LTs (B4 and C4) and PGs (E2 and F2alpha) and EDN-1 in the medium (EIA) were evaluated. RESULTS: We received immortalized luteal endothelial cell line (EnCL-1). Cytokines did not change EnCL-1 cell viability but increased mRNA expression of LTC4S, LTA4H, PGE2 and PGF2alpha synthases and EDN-1. EDN-1/2/3, LTC4 and PGF2alpha synthases protein expression were elevated in the presence of TNFalpha/IFNgamma, and accompanied by increased EDN-1, LTC4 and PGF2alpha secretion. Cytokines had no effect on PGES and LTA4H protein expression, and PGE2 and LTB4 release. CONCLUSIONS: TNFalpha and IFNgamma modulate EnCL-1 cell function. Moreover, established EnCL-1 cell line appears to be a good model for investigating the molecular mechanisms related to cytokines action and aa metabolites production in cattle.

Biomechanical Aspects of the Foot Arch, Body Balance and Body Weight Composition of Boys Training Football.

Background: The aim of the study is to assess the body balance and podological parameters and body composition of young footballers in the context of the control of football training. Methods: The study examined the distribution of the pressure of the part of the foot on the ground, the arch of the foot, and the analysis of the body composition of the boys. The pressure center for both feet and the whole body was also examined. The study involved 90 youth footballers from Olsztyn and Barczewo in three age groups: 8-10 years, 11-13 years old, and 14-16 years. The study used the Inbody 270 body composition analyzer and the EPSR1, a mat that measures the pressure distribution of the feet on the ground. Results: The results showed statistically significant differences in almost every case for each area of the foot between the groups of the examined boys. The most significant differences were observed for the metatarsal area and the left heel. In the case of stabilization of the whole body, statistically significant differences were noted between all study groups. In the case of the body composition parameters, in the examined boys, a coherent direction of changes was noticed for most of them. The relationships and correlations between the examined parameters were also investigated. The significance level in the study was set at p < 0.05. Conclusions: Under the training rigor, a statistically significant increase in stability was observed with age. The total length of the longitudinal arch of both feet of the examined boys showed a tendency to flatten in direct proportion to the age of the examined boys. Mean values of the body composition parameters reflect changes with the ontogenetic development, basic somatic parameters (body height and weight) and training experience, and thus with the intensity and volume of training. This indicates a correct training process that does not interfere with the proper development of the body in terms of tissue and biochemical composition.

Adipose-Derived Stromal/Stem Cells from Large Animal Models: from Basic to Applied Science.

Adipose-derived stem cells (ASCs) isolated from domestic animals fulfill the qualitative criteria of mesenchymal stem cells, including the capacity to differentiate along multiple lineage pathways and to self-renew, as well as immunomodulatory capacities. Recent findings on human diseases derived from studying large animal models, have provided evidence that administration of autologous or allogenic ASCs can improve the process of healing. In a narrow group of large animals used in bioresearch studies, pigs and horses have been shown to be the best suited models for study of the wound healing process, cardiovascular and musculoskeletal disorders. To this end, current literature demonstrates that ASC-based therapies bring considerable benefits to animal health in both spontaneously occurring and experimentally induced clinical cases. The purpose of this review is to provide an overview of the diversity, isolation, and characterization of ASCs from livestock. Particular attention has been paid to the functional characteristics of the cells that facilitate their therapeutic application in large animal models of human disease. In this regard, we describe outcomes of ASCs utilization in translational research with pig and horse models of disease. Furthermore, we evaluate the current status of ASC-based therapy in veterinary practice, particularly in the rapidly developing field of equine regenerative medicine. In conclusion, this review presents arguments that support the relevance of animal ASCs in the field of regenerative medicine and it provides insights into the future perspectives of ASC utilization in animal husbandry.

Distribution of feet pressure on ground and maintaining body balance among 8-10-year-old children with and without external load application.

PURPOSE: The aim of this study was to analyse the impact of applying an external load on the distribution of pressure on the plantar side of the foot and maintaining body balance, using the podobarographic platform. METHODS: The study was conducted on 130 school children aged 8-10: girls (n = 68, body mass = 22.8 ± 6.0 kg, body height = 129.3 ± 7.5 cm) and boys (n = 62, body mass = 31.1 ± 6.5 kg, body height 134.4 ± 7.3 cm). The study involved 2 trials. At first, children stood on the platform assuming a natural position. Then, they put on a 5-kg backpack and stood on the platform once more. RESULTS: The results indicate that after backpack loading, for the total research group of girls and boys, statistically significant differences were found in the distribution of foot force on the ground in the left forefoot ( p = 0.008), metatarsus ( p = 0.000) and heel areas ( p = 0.002). While in the right foot, these differences were noted for the forefoot ( p = 0.024) and metatarsus ( p = 0.000). The results of balance testing were also statistically significant. They concerned measurements of the body barycentre area (cop-bars p = 0.003), the barycentre area of the left foot (l-bars p = 0.034) and the parameter comparing distance to surface ratio (cop-lsf p = 0.000). CONCLUSIONS: It may be concluded that prolonged overloading with backpacks affects movement patterns, which may further lead to the acquisition and consolidation of postural defects.

The Transcription Factor FOXN1 Regulates Skin Adipogenesis and Affects Susceptibility to Diet-Induced Obesity.

FOXN1, a transcription factor expressed in the epidermis, regulates keratinocyte differentiation and participates in skin wound healing. In this study, we explored the impact of FOXN1 insufficiency on diet-stimulated weight gain and dermal white adipose tissue regulation in the intact and wounded skin of FOXN1eGFP/+ (heterozygotes, FOXN1-insufficient) mice in the context of age and diet. The results showed that on a high-fat diet, FOXN1eGFP/+ mice gained significantly less body weight than their FOXN1+/+ counterparts (FOXN1-sufficient mice). The intact and wounded skin of FOXN1eGFP/+ mice displayed abrogated expression of the master regulators of adipogenesis, PPARγ, FABP4, and leptin, which decreased with age in FOXN1+/+ mice. FOXN1 insufficiency also resulted in a decreased percentage of adipocyte-committed precursor cells (CD24+) in the skin. The proadipogenic pathway genes Bmp2, Igf2, and Mest showed a gradual decrease in expression that accompanied the gradual inactivation of FOXN1 in the skin of FOXN1+/+, FOXN1eGFP/+, and FOXN1eGFP/eGFP (lack of FOXN1) mice. Bone morphogenetic protein 2 and insulin-like growth factor 2 signals colocalized with FOXN1-eGFP in the epidermis and in hair follicles. These data demonstrated that FOXN1 initiates the cascade of adipogenic signaling that regulates skin homeostasis and wound healing and affects susceptibility to diet-induced obesity.

The effect of hypoxia on the proteomic signature of pig adipose-derived stromal/stem cells (pASCs).

Human adipose-derived stem cells (ASCs) have potential to improve wound healing; however, their equivalents from domestic animals have received less attention as an alternative cell-based therapy for animals or even humans. Hypoxia is essential for maintaining stem cell functionality in tissue-specific niches. However, a cellular response to low oxygen levels has not been demonstrated in pig ASCs. Hence, the goal of our study was to characterize ASCs isolated from the subcutaneous fat of domestic pigs (pASCs) and examine the effect of hypoxia on their proteome and functional characteristics that might reproduce pASCs wound healing ability. Analysis of immunophenotypic and functional markers demonstrated that pASCs exhibited characteristics of mesenchymal stem cells. Proteomic analysis revealed 70 differentially abundant proteins between pASCs cultured under hypoxia (1% O2) or normoxia (21% O2). Among them, 42 proteins were enriched in the cells exposed to low oxygen, whereas 28 proteins showed decrease expression following hypoxia. Differentially expressed proteins were predominantly involved in cell metabolism, regulation of focal and intracellular communication, and attributed to wound healing. Functional examination of hypoxic pASCs demonstrated acquisition of contractile abilities in vitro. Overall, our results demonstrate that hypoxia pre-conditioning impacts the pASC proteome signature and contractile function in vitro and hence, they might be considered for further cell-based therapy study on wound healing.