Secretory Activity Research Articles

Morphology of the epididymal duct of the domestic pig (Sus scrofa domesticus).

The study provides a general overview of the morphology of the epididymal duct in pigs. Four epididymides from two sexually mature boars were dissected into 32 segments and examined histologically. Duct lumen and wall thickness were measured and relative surface area of different components was assessed by Chalkley's random hit method. The epithelial lining was characterized at X1000. Lumen diameter and wall thickness of efferent ductules averaged 177 and 30 µm, respectively. Of the epididymal duct from caput to distal corpus the luminal diameter was 332 µm, with a narrower section in the proximal corpus. Wall thickness averaged 70 µm. In the cauda, luminal diameter and wall thickness increased to 717and 751 µm, respectively. The epithelial lining of the efferent ductules consists of a single layer of columnar cells with average height 21 µm. The lining of the epididymal duct consists of ciliated, pseudo-stratified columnar epithelium composed of "basal cells" and "principal cells." Particularly tall principal cells (96 µm) were found in the proximal caput. Height decreased to 40 µm at the distal cauda. Microvilli from principal cells were 14-17 µm long in the distal caput but decreased to 5 µm in the distal cauda. The epithelial lining was folded in the proximal caput and more so in the distal cauda. Secretory granules (epididymosomes) were present in small amounts in efferent ductules and epididymal duct; the largest quantities occurred in the distal cauda. Leukocytes were present throughout the duct, albeit in insignificant numbers. Chalkley's random hit method showed rapid spermatozoan transport through efferent ductules and proximal caput in large amounts of fluid. Sperm concentration increased due to fluid resorption in the proximal caput, was highest from caput flexure to proximal cauda and decreased at the caudal flexure, indicating secretory activity.

Dynamics of the Secretory Activity of the Female Moose Mammary Gland During the First Lactation at the Sumarokovskaya Moose Farm

For the first time, comprehensive research have been conducted to study the morphology and function of the mammary gland of domesticated first-calf moose. The mammary gland is an organ of dense consistency, consisting of four grouped hills divided into left and right halves, in which, in turn, the anterior and posterior lobes are distinguished, consisting of glandular (62.8%) and connective tissues (37.2%). Data on the secretory function duration, which varies from 120 to 42 days, and the productive qualities of the mammary gland (175.7 and 20.3 liters, respectively) in female moose with different activities of coming to the farm for milking are presented and systematized. The clarified features of the activity of the secretory system of the mammary gland of the moose-heifer represent the “standard.” Thus, they are essential to analyze species characteristics, comparative morphology, and physiology of lactation as a standard criterion for improving animal productivity at the stage of domestication, as well as to organize measures for the prevention of breast diseases in moose. The object of the study is elk cows of the first lactation (n= 221 heads). Depending on the activity of moose coming to the farm during lactation, they were divided into four groups: 1st - with 100% activity, 2nd – 99-80%, 3rd - 79-50%, and 4th – with 49% activity or less. The scientific information presented in the paper was obtained using morphological, histological, zootechnical, and statistical methods. Morphometric analysis showed the ratio of parenchyma and stroma in the mammary gland in domesticated moose 1.7:1. The glandular apparatus of the udder in the studied female moose secreted significantly higher in animals of the first (duration of lactation 120 days, lactation milk yield – 175.7±3.36 l) and the second groups (109 days and milk yield 122.9 ± 2.31 l) compared to the animals of the third (96 days and a yield of 80.4 ± 3.77 l) and fourth (42 days and a yield of 20.3 ± 1.06 l) groups. The analysis of the secretory activity of the mammary gland in female moose with different activities of coming to milking during the first lactation showed that domestication is a complex and lengthy process. There is still a long and purposeful work to be done to improve the issues of morphology, physiology of the lactation process, and regulation of growth and development of the mammary gland. Moreover, it is essential to organize and improve measures for the raising of moose replacements to transfer them to a milking herd. This will not only domesticate the moose on the farm and strengthen their attachment to humans but also prepare the animals for the upcoming productive period.

Specifics of Cryopreservation of Hydrogel Biopolymer Scaffolds with Encapsulated Mesenchymal Stem Cells.

The demand for regenerative medicine products is growing rapidly in clinical practice. Unfortunately, their use has certain limitations. One of these, which significantly constrains the widespread distribution and commercialization of such materials, is their short life span. For products containing suspensions of cells, this issue can be solved by using cryopreservation. However, this approach is rarely used for multicomponent tissue-engineered products due to the complexity of selecting appropriate cryopreservation protocols and the lack of established criteria for assessing the quality of such products once defrosted. Our research is aimed at developing a cryopreservation protocol for an original hydrogel scaffold with encapsulated MSCs and developing a set of criteria for assessing the quality of their functional activity in vitro. The scaffolds were frozen using two alternative types of cryocontainers and stored at either -40 °C or -80 °C. After cryopreservation, the external state of the scaffolds was evaluated in addition to recording the cell viability, visible changes during subsequent cultivation, and any alterations in proliferative and secretory activity. These observations were compared to those of scaffolds cultivated without cryopreservation. It was shown that cryopreservation at -80 °C in an appropriate type of cryocontainer was optimal for the hydrogels/adipose-derived stem cells (ASCs) tested if it provided a smooth temperature decrease during freezing over a period of at least three hours until the target values of the cryopreservation temperature regimen were reached. It was shown that evaluating a set of indicators, including the viability, the morphology, and the proliferative and secretory activity of the cells, enables the characterization of the quality of a tissue-engineered construct after its withdrawal from cryopreservation, as well as indicating the effectiveness of the cryopreservation protocol.

Boosting anaerobic digestion of long chain fatty acid with microbial electrolysis cell combining metal organic framework as cathode: Biofilm construction and metabolic pathways

The role of metal organic framework (MOF) modified cathode in promoting long chain fatty acid (LCFA) methanation was identified in microbial electrolysis cell coupled anaerobic digestion (MEC-AD) system. The maximum methane production rate of MEC-AD-MOF achieved 49.8 ± 3.4 mL/d, which increased by 41 % compared to MEC-AD-C. The analysis of bio-cathode biofilm revealed that microbial activity, distribution, population, and protein secretion prompted by MOF cathode, which in turn led to an acceleration of electron transfer between the cathode and microbes. Specifically, the relative abundance of acetate-oxidizing bacterium (Mesotoga) in MEC-AD-MOF was 1.5–3.6 times higher than that in MEC-AD-C, with a co-metabolized enrichment of Methanobacterium. Moreover, MOF cathode reinforced LCFA methanation by raising the relative abundance of genes coded key enzymes involved in CO2-reducing pathway, and elevating the tolerance of microbes to LCFA inhibition. These results indicate that MOF can enhance biofilm construction in MEC-AD, thereby improving the treatment performance of lipid wastewater.

Effect of running exercise training on inflammatory mediators and cytokines expression in testicular tissue; effect of exercise intensity

The aim of this study is to investigate the impact of running exercise training protocols (ETPs) with varying intensities on inflammatory responses, with a specific focus on the interactions between inflammatory mediators, cytokines, and Leydig cell steroidogenic activity, as well as testosterone secretion. To this end, 24 Wistar rats were subdivided into sedentary control, low (LICT), moderate (MICT), and high (HICT) intensity continuous running ETP groups. After 8 weeks, the expression levels of Toll-like receptor-4 (TLR-4), nuclear factor-kappa-B (NF-KB), interleukin-6 (IL-6), interleukin-10 (IL-10), Tumor necrosis factor alpha (TNF-α), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and the testicular nitric oxide (NO) content were assessed and compared between groups. Moreover, the mean distributions of Leydig cells/mm2 of interstitial connective tissue, their steroidogenic activity, and serum level of testosterone were assessed. The LICT did not show any significant (p > 0.05) change in the expression levels of all aforementioned biomarkers. In contrast, both the MICT and HICT groups demonstrated a significant (p < 0.05) increase in the expression levels of TLR-4, NFK-B, IL-6, TNF-α, iNOS, and COX-2 at both the mRNA and protein levels. The testicular NO has increased in HICT and MICT groups. Despite a decrease in the distribution of Leydig cells in both the MICT and HICT groups, the HICT group exhibited a significant (p < 0.05) reduction in Leydig cell steroidogenic activity and serum testosterone levels. In conclusion, our findings revealed that ETPs can influence Leydig cell steroidogenic activity and testosterone secretion, contingent on their intensity. These effects are attributed to alterations in the expression levels of pro-inflammatory mediators and cytokines.

Localization, traffic and function of Rab34 in adipocyte lipid and endocrine functions

BackgroundExcessive lipid accumulation in the adipose tissue in obesity alters the endocrine and energy storage functions of adipocytes. Adipocyte lipid droplets represent key organelles coordinating lipid storage and mobilization in these cells. Recently, we identified the small GTPase, Rab34, in the lipid droplet proteome of adipocytes. Herein, we have characterized the distribution, intracellular transport, and potential contribution of this GTPase to adipocyte physiology and its regulation in obesity.Methods3T3-L1 and human primary preadipocytes were differentiated in vitro and Rab34 distribution and trafficking were analyzed using markers of cellular compartments. 3T3-L1 adipocytes were transfected with expression vectors and/or Rab34 siRNA and assessed for secretory activity, lipid accumulation and expression of proteins regulating lipid metabolism. Proteomic and protein interaction analyses were employed for the identification of the Rab34 interactome. These studies were combined with functional analysis to unveil the role played by the GTPase in adipocytes, with a focus on the actions conveyed by Rab34 interacting proteins. Finally, Rab34 regulation in response to obesity was also evaluated.ResultsOur results show that Rab34 localizes at the Golgi apparatus in preadipocytes. During lipid droplet biogenesis, Rab34 translocates from the Golgi to endoplasmic reticulum-related compartments and then reaches the surface of adipocyte lipid droplets. Rab34 exerts distinct functions related to its intracellular location. Thus, at the Golgi, Rab34 regulates cisternae integrity as well as adiponectin trafficking and oligomerization. At the lipid droplets, this GTPase controls lipid accumulation and lipolysis through its interaction with the E1-ubiquitin ligase, UBA1, which induces the ubiquitination and proteasomal degradation of the fatty acid transporter and member of Rab34 interactome, FABP5. Finally, Rab34 levels in the adipose tissue and adipocytes are regulated in response to obesity and related pathogenic insults (i.e., fibrosis).ConclusionsRab34 plays relevant roles during adipocyte differentiation, including from the regulation of the oligomerization (i.e., biological activity) and secretion of a major adipokine with insulin-sensitizing actions, adiponectin, to lipid storage and mobilization from lipid droplets. Rab34 dysregulation in obesity may contribute to the altered adipokine secretion and lipid metabolism that characterize adipocyte dysfunction in conditions of excess adiposity.

How osteons form: A quantitative hypothesis-testing analysis of cortical pore filling and wall asymmetry

Osteon morphology provides valuable information about the interplay between different processes involved in bone remodelling. The correct quantitative interpretation of these morphological features is challenging due to the complexity of interactions between osteoblast behaviour, and the evolving geometry of cortical pores during pore closing. We present a combined experimental and mathematical modelling study to provide insights into bone formation mechanisms during cortical bone remodelling based on histological cross-sections of quiescent human osteons and hypothesis-testing analyses. We introduce wall thickness asymmetry as a measure of the local asymmetry of bone formation within an osteon and examine the frequency distribution of wall thickness asymmetry in cortical osteons from human iliac crest bone samples from women 16–78 years old. Our measurements show that most osteons possess some degree of asymmetry, and that the average degree of osteon asymmetry in cortical bone evolves with age. We then propose a comprehensive mathematical model of cortical pore filling that includes osteoblast secretory activity, osteoblast elimination, osteoblast embedment as osteocytes, and osteoblast crowding and redistribution along the bone surface. The mathematical model is first calibrated to symmetric osteon data, and then used to test three mechanisms of asymmetric wall formation against osteon data: (i) delays in the onset of infilling around the cement line; (ii) heterogeneous osteoblastogenesis around the bone perimeter; and (iii) heterogeneous osteoblast secretory rate around the bone perimeter. Our results suggest that wall thickness asymmetry due to off-centred Haversian pores within osteons, and that nonuniform lamellar thicknesses within osteons are important morphological features that can indicate the prevalence of specific asymmetry-generating mechanisms. This has significant implications for the study of disruptions of bone formation as it could indicate what biological bone formation processes may become disrupted with age or disease.

Biological properties and characterization of several variations of a clinical human plasma-based skin substitute model and its manufacturing process.

Human plasma is a natural biomaterial that due to their protein composition is widely used for the development of clinical products, especially in the field of dermatology. In this context, this biomaterial has been used as a scaffold alone or combined with others for the development of cellular human plasma-based skin substitutes (HPSSs). Herein, the biological properties (cell viability, cell metabolic activity, protein secretion profile and histology) of several variations of a clinical HPSS model, regarding the biomaterial composition (alone or combined with six secondary biomaterials - serine, fibronectin, collagen, two types of laminins and hyaluronic acid), the cellular structure (trilayer, bilayer, monolayer and control without cells) and their skin tissue of origin (abdominal or foreskin cells) and the manufacturing process [effect of partial dehydration process in cell viability and comparison between submerged (SUB) and air/liquid interface (ALI) methodologies] have been evaluated and compared. Results reveal that the use of human plasma as a main biomaterial determines the in vitro properties, rather than the secondary biomaterials added. Moreover, the characteristics are similar regardless of the skin cells used (from abdomen or foreskin). However, the manufacture of more complex cellular substitutes (trilayer and bilayer) has been demonstrated to be better in terms of cell viability, metabolic activity and wound healing protein secretion (bFGF, EGF, VEGF-A, CCL5) than monolayer HPSSs, especially when ALI culture methodology is applied. Moreover, the application of the dehydration, although required to achieve an appropriate clinical structure, reduce cell viability in all cases. These data indicate that this HPSS model is robust and reliable and that the several subtypes here analysed could be promising clinical approaches depending on the target dermatological disease.

ULTRASTRUCTURAL CHANGES IN RATS SKIN SEBOCYTES DURING EXPERIMENTAL CHRONODESTRUCTION

Purpose of the study ultrastructural changes in rat skin sebocytes during experimental chronodestruction. In the experiments, 22 white outbred male rats with a body weight of 170-220 g were used. The experimental animals, in accordance with the experimental design, were randomly divided into 3 groups: group 1 - intact (n = 8) - animals under standard fixed conditions lighting (12 hours light/12 hours dark); group 2 (n=8) – animals with simulated light deprivation in conditions of round-the-clock darkness (24-hour darkness) for 21 days; group 3 (n=6) – animals with simulated dark deprivation under conditions of round-the-clock lighting (24 hours light) for 21 days. For morphometric assessment, 30 non-overlapping fields of view were analyzed in each preparation at a magnification of 100,000. The number of mitochondria and lysosomes was calculated per 100 μm2 area of the cytoplasm of cells of a certain morphological type. Using the ImageScopeM application program, the average cross-sectional area of mitochondria (μm2) and the average relative electron density of their matrix were determined. During chronodestruction, significant ultrastructural changes are observed in sebocytes, which indicate a change in the functional activity of the cells. During dark deprivation, the functional activity of sebocytes increases, and this, in turn, leads to a significant change in their secretory activity. At the same time, light deprivation leads to an increase in proliferative activity in the sebaceous gland with a simultaneous change in the chemical composition of sebum (polymorphism of granules).

Endogenous bystander killing mechanisms enhance the activity of novel FAP-specific CAR-Tcells against glioblastoma.

CAR-T cells are being investigated as a novel immunotherapy for glioblastoma, but clinical success has been limited. We recently described fibroblast activation protein (FAP) as an ideal target antigen for glioblastoma immunotherapy, with expression on both tumor cells and tumor blood vessels. However, CAR-T cells targeting FAP have never been investigated as a therapy for glioblastoma. We generated a novel FAP targeting CAR with CD3ζ and CD28 signalling domains and tested the resulting CAR-T cells for their lytic activity and cytokine secretion function invitro (using real-time impedance, flow cytometry, imaging and bead-based cytokine assays), and in vivo (using a xenograft mimicking the natural heterogeneity of human glioblastoma). FAP-CAR-T cells exhibited target specificity against model cell lines and potent cytotoxicity against patient-derived glioma neural stem cells, even when only a subpopulation expressed FAP, indicating a bystander killing mechanism. Using co-culture assays, we confirmed FAP-CAR-T cells mediate bystander killing of antigen-negative tumor cells, but only after activation by FAP-positive target cells. This bystander killing was at least partially mediated by soluble factors and amplified by IL-2 which activated the non-transduced fraction of the CAR-T product. Finally, a low dose of intravenously administered FAP-CAR-T cells controlled, without overt toxicity, the growth of subcutaneous tumors created using a mixture of antigen-negative and antigen-positive glioblastoma cells. Our findings advance FAP as a leading candidate for clinical CAR-T therapy of glioblastoma and highlight under-recognised antigen nonspecific mechanisms that may contribute meaningfully to the antitumor activity of CAR-T cells.

Exhausted signature and regulatory network of NK cells in myasthenia gravis.

NK cells are dysfunctional in myasthenia gravis (MG), but the mechanism is unclear. This study aims to measure associations and underlying mechanisms between the NK cells and the development of MG. Twenty healthy controls (HCs) and 53 MG patients who did not receive glucocorticoids and immunosuppressants were collected. According to the Myasthenia Gravis Foundation of America (MGFA) classification, MG patients were categorized into MGFA I group (n = 18) and MGFA II-IV group (n = 35). Flow cytometry, cell sorting, ELISA, mRNA-sequencing, RT-qPCR, western blot, and cell culture experiments were performed to evaluate the regulatory mechanism of exhausted NK cells. Peripheral NK cells in MGFA II-IV patients exhibit exhausted phenotypes than HCs, marked by the dramatic loss of total NK cells, CD56dimCD16- NK cells, elevated PD1 expression, reduced NKG2D expression, impaired cytotoxic activity (perforin, granzyme B, CD107a) and cytokine secretion (IFN-γ). Plasma IL-6 and IL-21 are elevated in MG patients and mainly derived from the aberrant expansion of monocytes and Tfh cells, respectively. IL-6/IL-21 cooperatively induced NK-cell exhausted signature via upregulating SOCS2 and inhibiting the phosphorylation of STAT5. SOCS2 siRNA and IL-2 supplement attenuated the IL-6/IL-21-mediated alteration of NK-cell phenotypes and function. Inhibition of IL-6/IL-21/SOCS2/STAT5 pathway and recovery of NK-cell ability to inhibit autoimmunity may be a new direction in the treatment of MG.

Extracellular electrophysiology on clonal human β-cell spheroids.

Pancreatic islets are important in nutrient homeostasis and improved cellular models of clonal origin may very useful especially in view of relatively scarce primary material. Close 3D contact and coupling between β-cells are a hallmark of physiological function improving signal/noise ratios. Extracellular electrophysiology using micro-electrode arrays (MEA) is technically far more accessible than single cell patch clamp, enables dynamic monitoring of electrical activity in 3D organoids and recorded multicellular slow potentials (SP) provide unbiased insight in cell-cell coupling. We have therefore asked whether 3D spheroids enhance clonal β-cell function such as electrical activity and hormone secretion using human EndoC-βH1, EndoC-βH5 and rodent INS-1 832/13 cells. Spheroids were formed either by hanging drop or proprietary devices. Extracellular electrophysiology was conducted using multi-electrode arrays with appropriate signal extraction and hormone secretion measured by ELISA. EndoC-βH1 spheroids exhibited increased signals in terms of SP frequency and especially amplitude as compared to monolayers and even single cell action potentials (AP) were quantifiable. Enhanced electrical signature in spheroids was accompanied by an increase in the glucose stimulated insulin secretion index. EndoC-βH5 monolayers and spheroids gave electrophysiological profiles similar to EndoC-βH1, except for a higher electrical activity at 3 mM glucose, and exhibited moreover a biphasic profile. Again, physiological concentrations of GLP-1 increased AP frequency. Spheroids also exhibited a higher secretion index. INS-1 cells did not form stable spheroids, but overexpression of connexin 36, required for cell-cell coupling, increased glucose responsiveness, dampened basal activity and consequently augmented the stimulation index. In conclusion, spheroid formation enhances physiological function of the human clonal β-cell lines and these models may provide surrogates for primary islets in extracellular electrophysiology.

β-Cell Heterogeneity and Plasticity.

The existence of functionally diverse and plastic β cells in islets of Langerhans has been reported since the 1980s. Recently, high-resolution technologies have advanced our understanding of β-cell heterogeneity and plasticity. Here, we define plasticity broadly as dynamic changes in cellular phenotypes and heterogeneity as differences in cellular behaviors. Individual β cells react differently to environmental challenges and act together to maintain β-cell mass and glucose homeostasis within a narrow range of 70-140mg/dL. During the progress of diabetes, this elaborate balance is disrupted, and a lack of β-cell compensation leads to dysregulated blood glucose. In this chapter, we assess β-cell stress that instigates increased β-cell heterogeneity and adaptive β-cell responses such as proliferation, dedifferentiation, maturity, and insulin secretion. We also discuss the maturity, electrical activity, and insulin secretion of well-characterized β-cell subgroups. Finally, we touch upon the plasticity of other non-β pancreatic cells and their cooperation with β cells to maintain homeostasis.

NGF effects promote the maturation of rat pancreatic beta cells by regulating GLUT2 levels and distribution, and glucokinase activity.

The nerve growth factor (NGF) participates in cell survival and glucose-stimulated insulin secretion (GSIS) processes in rat adult beta cells. GSIS is a complex process in which metabolic events and ionic channel activity are finely coupled. GLUT2 and glucokinase (GK) play central roles in GSIS by regulating the rate of the glycolytic pathway. The biphasic release of insulin upon glucose stimulation characterizes mature adult beta cells. On the other hand, beta cells obtained from neonatal, suckling, and weaning rats are considered immature because they secrete low levels of insulin and do not increase insulin secretion in response to high glucose. The weaning of rats (at postnatal day 20 in laboratory conditions) involves a dietary transition from maternal milk to standard chow. It is characterized by increased basal plasma glucose levels and insulin levels, which we consider physiological insulin resistance. On the other hand, we have observed that incubating rat beta cells with NGF increases GSIS by increasing calcium currents in neonatal cells. In this work, we studied the effects of NGF on the regulation of cellular distribution and activity of GLUT2 and GK to explore its potential role in the maturation of GSIS in beta cells from P20 rats. Pancreatic islet cells from both adult and P20 rats were isolated and incubated with 5.6 mM or 15.6 mM glucose with and without NGF for 4 hours. Specific immunofluorescence assays were conducted following the incubation period to detect insulin and GLUT2. Additionally, we measured glucose uptake, glucokinase activity, and insulin secretion assays at 5.6 mM or 15.6 mM glucose concentrations. We observed an age-dependent variation in the distribution of GLUT2 in pancreatic beta cells and found that glucose plays a regulatory role in GLUT2 distribution independently of age. Moreover, NGF increases GLUT2 abundance, glucose uptake, and GSIS in P20 beta cells and GK activity in adult beta cells. Our results suggest that besides increasing calcium currents, NGF regulates metabolic components of the GSIS, thereby contributing to the maturation process of pancreatic beta cells.

МЕТАБОЛИЧЕСКИЕ РЕАКЦИИ ОРГАНИЗМА ЗДОРОВОГО ЧЕЛОВЕКА В ДИНАМИКЕ ЭКСПЕРИМЕНТА SIRIUS-21

The SIRIUS-21 study was designed as a 240-day isolation of healthy volunteers in controlled environment simulating a piloted mission to the Moon including operator's activities in lunar orbit and on the surface. The investigation involved an international crew of two women and 3 men at the age of 29 to 44 years. The object was metabolic reactions of healthy bodies to the simulated conditions. Crew members demonstrated a moderate stress reaction characterized by inhibition of the gastrointestinal tract secretory activity, reduction of glycolysis intensity, persistent hypercalcemia, a more intensive creatine kinase system functioning as a buffer, deficit of macroergic compounds, magnesium and essential microelements. On day 7 post isolation, the stress symptoms showed a partial leveling. Hard physical training had both the positive and negative effects on metabolism dynamics. The preventive purpose of training was to normalize the content of essential microelements in blood and suppress atherogenesis; however, training had also the negative effect of inducing hypohydration. In future isolation studies it would be appropriate to optimize training protocols and enhance the program of countermeasures to the negative effects of simulated factors.

Tumor necrosis factor-alpha as a modulator of secretory activity and protein cargo of extracellular vesicles in mesenchymal stromal cells

Tumor necrosis factor-alpha as a modulator of secretory activity and protein cargo of extracellular vesicles in mesenchymal stromal cells

Exercise During Pregnancy

It has been established that the synthetic peptide LKEKK affects the ability of IL-12 to regulate the secretory activity of keratinocytes obtained from human psoriatic skin: in the concentration range of 100-1000 nM the peptide increases in a dose-dependent manner the production of IL-10 and IFN-γ as well as suppression of the production of IL- 17 induced by IL-12 in vitro. The peptide with the inverted sequence KKEKL was inactive, which indicates the high specificity of the peptide LKEKK action. An in vivo study of the peptide LKEKK activity in a model of imiquimod (IMQ)-induced psoriasis in mice showed that its daily application to the ear (150 500 µg) together with Aldara cream containing 5% IMQ for 6 days significantly suppresses the development of the inflammatory process.

GPR55 inhibits the pro-adipogenic activity of anandamide in human adipose stromal cells

The endocannabinoid anandamide (AEA) stimulates adipogenesis via the cannabinoid receptor CB1 in adipose stromal cells (ASCs). However, AEA interacts also with nonclassical cannabinoid receptors, including transient receptor potential cation channel (TRPV)1 and G protein-coupled receptor (GPR)55. Their roles in AEA mediated adipogenesis of human ASCs have not been investigated.We examined the receptor-expressions by immunostaining on human ASCs and tested their functionality by measuring the expression of immediate early genes (IEGs) related to the transcription factor-complex AP-1 upon exposition to receptor agonists. Cells were stimulated with increasing concentrations of specific ligands to investigate the effects on ASC viability (proliferation and metabolic activity), secretory activity, and AEA mediated differentiation.ASCs expressed both receptors, and their activation suppressed IEG expression. TRPV1 did not affect viability or cytokine secretion. GPR55 decreased proliferation, and it inhibited the release of hepatocyte growth factor. Blocking GPR55 increased the pro-adipogenic activity of AEA. These data suggest that GPR55 functions as negative regulator of cannabinoid mediated pro-adipogenic capacity in ASCs.

Inferring secretory and metabolic pathway activity from omic data with secCellFie

Understanding protein secretion has considerable importance in biotechnology and important implications in a broad range of normal and pathological conditions including development, immunology, and tissue function. While great progress has been made in studying individual proteins in the secretory pathway, measuring and quantifying mechanistic changes in the pathway's activity remains challenging due to the complexity of the biomolecular systems involved. Systems biology has begun to address this issue with the development of algorithmic tools for analyzing biological pathways; however most of these tools remain accessible only to experts in systems biology with extensive computational experience. Here, we expand upon the user-friendly CellFie tool which quantifies metabolic activity from omic data to include secretory pathway functions, allowing any scientist to infer properties of protein secretion from omic data. We demonstrate how the secretory expansion of CellFie (secCellFie) can help predict metabolic and secretory functions across diverse immune cells, hepatokine secretion in a cell model of NAFLD, and antibody production in Chinese Hamster Ovary cells.

MAST CELLS IN THE MILIEU: A COMPARATIVE STUDY IN THE TONGUE AND LIVER

Mast Cells (MCs) have diverse roles in health and disease. These immune cells migrate in a tissue-specific manner to target sites where they can alert or amplify immune responses through exocytosis of their bioactive granules. Granule exocytosis can be triggered by both external immunological and non-immunological signals. Depending on the local milieu, MCs are highly heterogeneous cells with significant phenotypic plasticity and functional diversity. The aim of this study was to investigate differences in MC populations in the tongue and liver to advance the understanding of how organ-specific location may affect MC phenotypic variance and functions. MCs were investigated using light and electron microscopy in tissues obtained from rats housed in identical conditions. Connective tissue MCs were identified in both the tongue and liver and showed phenotypic differences in secretory activity and granule characteristics. These findings indicate that phenotypic variance depends on tissue location. This knowledge may help develop strategies for preventing or treating diseases.