Adaptive response triggered by the repeated SCUBA diving is reflected in cardiovascular, muscular, and immune biomarkers.

It has been shown that one recreational SCUBA (rSCUBA) diving session is sufficient to cause changes in plasma level of cardiovascular (CV) and muscular biomarkers. To explore whether repetitive rSCUBA diving triggers an adaptive response of the CV, muscular, and immune system, we measured the cardiac damage (NT-proBNP, hs-TnI, and CK-MB), muscle damage (myoglobin (Mb), galectin-3, CK, and LDH), vascular endothelial activation (ET-1 and VEGF), and inflammatory (leukocyte count (Lkc), CRP, and IL-6) biomarkers. A longitudinal intervention study included divers (N = 14) who conducted one dive per week over 5 weeks at the depth of 20-30 m for 30 min after a non-dive period of 5 months. The blood samples were collected before and after the first, third, and fifth dives and specific biomarkers were measured in plasma or serum by the standard laboratory methods. The concentrations of the majority of measured biomarkers increased after every single dive; the exception was ET-1 concentration that decreased. The cumulative effect of five dives has been reflected in diminishing changes in hs-TnI, Mb, galectin-3, ET-1, VEGF, and IL-6 levels, and more pronounced increases in NT-proBNP and hs-CRP levels. The median values of all measured biomarkers in all time points, except Mb, remained within the corresponding reference range. Repeatedly performed rSCUBA diving activates an adaptive response of the CV, muscular, and immune system that is reflected in changes in the specific biomarker concentration.

Galectin-3 and Cardiovascular Biomarkers Reflect Adaptation Response to Scuba Diving.

To understand better the adaptation response of the cardiovascular system (CVS) to self-contained underwater breathing apparatus (SCUBA) diving, Galectin-3 (Gal-3) and specific CVS biomarkers were measured in plasma of 16 male recreational divers before and after (30 min, 3 and 6 h) diving (total time of 30 min at 30 m depth) undertaken a after long non-dive period. The one-time SCUBA dive caused a significant increase in Gal-3, N-terminal prohormone of brain natriuretic peptide (NT-proBNP), high-sensitivity troponin-I (hs-TnI), and myoglobin immediately after diving. Whereas Gal-3 and myoglobin dropped down to the basal levels during the recovery period, NT-proBNP and hs-TnI concentration continued to increase. An immediate increase of vascular endothelial growth factor, detected immediately after diving, was followed by a significant decrease and return to the basal level, 3 and 6 h after diving, respectively. After a significant initial decrease, endothelin-1 increased during the recovery period, but did not return to the basal level. The observed changes in these biomarkers reflect comprehensive, but transient adaptation of CVS and muscular system to the specific environmental conditions during the SCUBA dive. Whether the recurrent activation of these adaption mechanisms due to repetitive dives has positive or negative effects on CVS remains to be elucidated.

Differential expression of heat shock proteins and activation of mitogen-activated protein kinases in A549 alveolar epithelial cells exposed to cigarette smoke extract.

NEW FINDINGS: What is the central question of this study? What is the effect of cigarette smoke on cell death, oxidative damage, expression of heat shock proteins (HSPs) and activation of mitogen-activated protein kinases (MAPKs) in A549 alveolar epithelial cells? What is the main finding and its importance? Cigarette smoke induces cytotoxicity and oxidative damage to A549 cells, increases expression of different HSPs and activates MAPK signalling pathways. This could be related to inflammatory response and apoptosis observed in lungs of patients with smoking-related diseases. ABSTRACT: Cigarette smoking is one of the main risk factors for development of chronic obstructive pulmonary disease (COPD). We previously reported that cigarette smoke (CS) induces damage to proteins and their ineffective degradation. Here, we hypothesize that CS could induce oxidative stress and cytotoxicity in lung epithelial cells through alterations of heat shock protein (HSP) expression and mitogen-activated protein kinase (MAPK) signalling pathways. We exposed A549 alveolar epithelial cells to various concentrations of cigarette smoke extract (CSE). Higher concentrations of CSE caused apoptosis of A549 cells after 4 h, while after 24 h cell viability was decreased, and lactate dehydrogenase in cell culture medium was increased as well as the number of necrotic cells. Concentrations of malondialdehyde (MDA) were elevated, while total thiol groups were decreased. Changes in the expression of HSPs (HSP70, HSP32 and HSP27) were time-dependent. After 6 h, CSE caused an increase in the expression of HSP70 and HSP32, while after 8 h all examined HSPs were up-regulated and remained increased up to 48 h. Treatment of A549 cells with CSE stimulated phosphorylation of extracellular signal-regulated kinase and p38 in a dose-dependent manner, while c-Jun N-terminal kinase activation was not detected. By using specific inhibitors, we demonstrated that MAPKs and HSPs interplay in CSE effects. In conclusion, our results show that MAPKs and HSPs are involved in the mechanism underlying CSE-induced cytotoxicity and oxidative damage to A549 alveolar epithelial cells. These processes could be related to inflammatory response and apoptosis observed in lungs of patients with smoking-related diseases, such as COPD.

Clinical and Biochemical Influence of Prostatic Stones.

INTRODUCTION: The study aimed to explore clinical influence of prostatic stones on lower urinary tract symptoms (LUTS), seminal plasma cytokines, and serum biomarkers. MATERIALS AND METHODS: A total of 70 men aged ≤50 years with LUTS divided into 2 groups: group with stones (GSt) and group without prostatic stones (GNoSt). All subjects completed the International Prostate Symptom Score (IPSS) questionnaire and National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) scoring questionnaire. Pre- and post-prostate massage test and uroflowmetry were performed. The serum concentration of total prostate specific antigen (PSA), free PSA, and free/total PSA (f/t PSA) ratio, seminal concentration of cytokines interleukin (IL)-1ß, IL-6, IL-8, IL-10, IL-12p70, and tumor necrosis factor-alpha were measured. RESULTS: GSt subjects had significantly more severe symptoms based on IPSS answers (p = 0.0289). All domains in NIH-CPSI scores were significantly higher in the GSt group: pain (p = 0.001), urinary symptoms (p = 0.023), quality of life (p = 0.008), and with overall (p = 0.003). GSt subjects also had significantly lower maximum urinary flow (Qmax; p = 0.011), lower f/t PSA ratio (p = 0.048), and higher concentration of IL-1ß (p = 0.011) and IL-8 (p = 0.001). CONCLUSIONS: Prostatic stones may influence the severity of LUTS and the symptoms of chronic prostatitis. They might reduce Qmax rate and lead to reduction of the f/t PSA ratio and produce more severe inflammation causing increased seminal concentration of IL-1ß and IL-8.

Recreational scuba diving: negative or positive effects of oxidative and cardiovascular stress?

Environmental conditions and increased physical activity during scuba diving are followed by increased production of free radicals and disturbed redox balance. Redox balance disorder is associated with damage of cellular components, changes of cellular signaling pathways and alterations of gene expression. Oxidative stress leads to increased expression of sirtuins (SIRTs), molecules which play an important role in the antioxidant defense, due to their sensitivity to the changes in the redox status and their ability to regulate redox homeostasis. These facts make SIRTs interesting to be considered as molecules affected by scuba diving and in that sense, as potential biomarkers of oxidative status or possible drug targets in reduction of reactive oxygen species (ROS) accumulation. In addition, SIRTs effects through currently known targets make them intriguing molecules which can act positively on health in general and whose expression can be induced by scuba diving.A demanding physical activity, as well as other circumstances present in scuba diving, has the greatest load on the cardiovascular function (CV). The mechanisms of CV response during scuba diving are still unclear, but diving-induced oxidative stress and the increase in SIRTs expression could be an important factor in CV adaptation. This review summarizes current knowledge on scuba diving-induced oxidative and CV stress and describes the important roles of SIRTs in the (patho)physiological processes caused by the redox balance disorder.

Galectin-3 endocytosis by carbohydrate independent and dependent pathways in different macrophage like cell types.

BACKGROUND: Galectin-3 (the Mac-2 antigen) is abundantly expressed in both macrophage like cells and certain non-macrophage cells. We have studied endocytosis of galectin-3 as one important step relevant for its function, and compared it between variants of a macrophage like cell line, and non-macrophage cells. METHODS: Endocytosis of galectin-3 was observed by fluorescence microscopy and measured by flow cytometry. The endocytosis mechanism was analysed using galectin-3 mutants, galectin-3 inhibitors and endocytic pathways inhibitors in the human leukaemia THP-1 cell line differentiated into naïve (M0), classical (M1) or alternatively activated (M2) macrophage like cells, and the non-macrophage cell lines HFL-1 fibroblasts and SKBR3 breast carcinoma. RESULTS: Galectin-3 endocytosis in non-macrophage cells and M2 cells was blocked by lactose and a potent galectin-3 inhibitor TD139, and also by the R186S mutation in the galectin-3 carbohydrate recognition domain (CRD). In M1 cells galectin-3 endocytosis could be inhibited only by chlorpromazine and by interference with the non-CRD N-terminal part of galectin-3. In all the cell types galectin-3 entered early endosomes within 5-10 min, to be subsequently targeted mainly to non-degradative vesicles, where it remained even after 24 h. CONCLUSIONS: Galectin-3 endocytosis in M1 cells is receptor mediated and carbohydrate independent, while in M2 cells it is CRD mediated, although the non-CRD galectin-3 domain is also involved. General significance The demonstration that galectin-3 endocytosis in M1 macrophages is carbohydrate independent and different from M2 macrophages and non-macrophage cells, suggests novel, immunologically significant interactions between phagocytic cells, galectin-3 and its ligands.

Galectin-1 and galectin-3 expression profiles in classically and alternatively activated human macrophages.

BACKGROUND: Galectins have been identified as modulators of many monocyte/macrophage functions. In the response to a wide range of environmental cues macrophages may exhibit different biochemical and biological characteristics, but two main subtypes, classically (M1) and alternatively (M2) activated macrophages have been recognized. To contribute to elucidation of role and regulation of galectin-1 and galectin-3 in differently activated macrophages we explored their expression profiles in these cells. METHODS: Human monocytes obtained from blood donors were differentiated into classically (M1) and alternatively (M2a/M2c) activated macrophages. Gene and protein expression levels of intra- and extracellular galectins were investigated by qRT-PCR, Western-blot, flow cytometry, and ELISA while cytokine and surface receptor expression profiling was performed by flow cytometry. RESULTS: Differentiation/polarization of human monocytes into classically (M1) and alternatively (M2a/M2c) activated macrophages was followed by profound changes of galectin-3 expression and its proteolytic cleavage. Expression and secretion of Gal-3 was tightly regulated and significantly differed among classically (M1) and alternatively (M2a/M2c) activated macrophages, while the differences of galectin-1 expression profiles were not as pronounced. Human monocytes exhibited high amount of free galectin-3 receptors, while on both types of activated macrophages were fully saturated. CONCLUSIONS: Galectin-3 is more distinctive descriptor of macrophages differentiation/activation than galectin-1. Its specific expression and secretion pattern in M1 vs. M2a/M2c macrophages contributes to better understanding of its role and regulation in these cells. GENERAL SIGNIFICANCE: Recognition of distinct galectin-1 and galectin-3 expression profiles in differently activated macrophages provides a new insight on biological characteristics of these cells and sheds a new light of galectin-3 as a modulator of individual macrophage subset. This article is part of a Special Issue entitled Glycoproteomics.

Aberrant glycosylation of Igg heavy chain in multiple myeloma.

Although the majority of eukaryotic proteins are glycosylated, there is a dearth of knowledge regarding protein sugar moieties and their changes in disease. Most multiple myeloma cases are characterized by production of monoclonal immunoglobulins (Ig). We studied galactosylation and sialylation of IgG heavy chains in 16 patients with IgG myeloma using lectin blotting and densitometry. In comparison to age and sex matched controls, galactosylation was reduced in multiple myeloma (median 317 vs. 362, range 153-410 vs. 309-447 relative units, p = 0.015, Student's t-test). Sialylation was stage dependent; samples from patients with stage IIA had lowest amounts of sialic acid, IIIA intermediate and IIIB highest (142.6 vs. 185.9 vs. 248.5 relative units, correlation coefficient r = 0.55). Both galactosylation and sialylation levels were independent of age, sex, treatment type, response to treatment, disease duration and IgG and b2 microglobulin concentration. These data indicate that multiple myeloma is characterized by aberrant immunoglobulin glycosylation.

Glycoscience -- a new frontier in rational drug design.

Glycans are the most abundant and most diverse biopolymers in nature. Because of their highly specific interactions with physiological receptors, they participate in many crucial biological processes. All these processes are potential targets for therapeutic intervention, and carbohydrate-based drugs are rapidly being taken up by the modern biotechnology and pharmaceutical industry. Recent developments in the field of glycobiology have overcome the problem of glycan analysis and synthesis; and many compounds based on carbohydrates are now in various stages of clinical trials. This article presents glycoproteins in a new light, as an important biopharmaceutical target, giving an overview of their potential use as therapeutic glycoproteins and proteoglycans, inflammation blockers, cancer therapeutics and vaccines, inhibitors of pathogenic microbes, viral inhibitors and potential aids in the treatment of lysosomal diseases, neurological diseases and transplantation rejection.

Galectin-3: an open-ended story.

Galectins, an ancient lectin family, are characterized by specific binding of beta-galactosides through evolutionary conserved sequence elements of carbohydrate-recognition domain (CRD). A structurally unique member of the family is galectin-3; in addition to the CRD it contains a proline- and glycine-rich N-terminal domain (ND) through which is able to form oligomers. Galectin-3 is widely spread among different types of cells and tissues, found intracellularly in nucleus and cytoplasm or secreted via non-classical pathway outside of cell, thus being found on the cell surface or in the extracellular space. Through specific interactions with a variety of intra- and extracellular proteins galectin-3 affects numerous biological processes and seems to be involved in different physiological and pathophysiological conditions, such as development, immune reactions, and neoplastic transformation and metastasis. The review attempts to summarize the existing information on structural, biochemical and intriguing functional properties of galectin-3.

Galectin-3 in macrophage-like cells exposed to immunomodulatory drugs.

During the last few decades, the effects of immunomodulatory drugs on numerous molecules and biological processes have been widely studied. Nevertheless, the relationship between immunomodulatory drugs and lectin expression/function is still to be elucidated. In this study, we used THP-1-derived macrophages to investigate the effects of non-steroidal anti-inflammatory drugs (aspirin and indomethacin) and glucocorticoids (hydrocortisone and dexamethasone) on galectin-3, a multifunctional beta-galactoside binding lectin, which in general acts as a strong pro-inflammatory signal. The results showed that all immunomodulatory drugs applied in clinically relevant doses affect both the gene (LGALS3) and protein expression level of galectin-3. The provoked changes on protein level are qualitatively and quantitatively different comparing to the effects on galectin-3 mRNA level, and depend on the differentiation state of the cell, drug type and applied concentration as well as on time of the exposure. Our data revealed galectin-3 as a new target molecule of immunomodulatory drugs, thus suggesting an additional pathway of their action on immune response.