Effects of recreational scuba diving on erythropoiesis-"normobaric oxygen paradox" or "plasma volume regulation" as a trigger for erythropoietin?

PURPOSE: Previous studies have shown an increase in erythrocyte lipid peroxidation and a decrease in red blood cell (RBC) count, hemoglobin, and hematocrit after only one recreational scuba diving session. The aim of this study was to examine the effect of repetitive scuba diving on RBC parameters and erythropoiesis. METHODS: Divers (N = 14) conducted one dive per week over 5 weeks at a depth of 20-30 m for 30 min. For measuring RBC parameters, erythropoietin, iron, and ferritin, blood samples were collected before and after the first, third, and fifth dive. RESULTS: Between pre- and post-dive results, a statistically significant increase in RBC count, hemoglobin, hematocrit, mean corpuscular volume (MCV), RBC distribution width (RDW), iron, and ferritin was observed. Analysis of the results between the first, third, and fifth dive showed that the erythropoietin increase at the third (pre-dive p = 0.009; post-dive p = 0.004) and fifth dive (pre-dive p < 0.001; post-dive p = 0.003) was not accompanied by changes in RBC count, hemoglobin, iron, and ferritin. In parallel, a continuous increase in hematocrit, MCV, and RDW was observed, whereas mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) decreased. CONCLUSIONS: Changes in RBC indices and EPO elevation indicate that the occasional switch from hyperoxia to normoxia or mechanisms for plasma volume regulation may be a step in the maintenance of erythropoiesis.

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.

Influence of hemolysis on clinical chemistry parameters determined with Beckman Coulter tests - detection of clinically significant interference.

The aim of this study was to examine the influence of hemolysis on 25 clinical chemistry parameters and to compare the resulting bias with clinically significant differences and the manufacturer's specifications. Using freeze-thawing of the treated blood aliquot of each subject (N = 17), four hemolysis levels were prepared with hemolysis index (HI) and hemoglobin concentration as follows: (+)=0.5-0.99 g/L, (2+)=1-1.99 g/L, (3+)=2-2.99 g/L and (4+)=3-4.99 g/L. All analytes were tested on the Beckman Coulter AU480 analyzer using proprietary reagents. It was considered that the interference was detected if the 95% confidence interval for mean differences (%) between hemolyzed and non-hemolyzed samples did not include zero. Clinically significant interference was judged against reference change value (RCV). Hemolysis interference was detected for: alpha-amylase, alkaline phosphatase (ALP), aspartate aminotransferase (AST), total and conjugated bilirubin, creatine kinase (CK), CK-MB, ɣ-glutamyltransferase (GGT), iron, lactate dehydrogenase (LD), magnesium, potassium, total protein and uric acid at HI=(1+); alanine aminotransferase (ALT) and phosphate at HI=(2+); urea at HI=(3+); albumin and cholinesterase at HI=(4+). Even at the greatest hemolysis degree, HI=(4+), no interference was detected for calcium, chloride, creatinine, C-reactive protein (CRP), glucose and sodium. Clinically significant difference was exceeded for LD at HI=(1+); CK-MB at HI=(2+); AST and potassium at HI=(3+); total bilirubin at HI=(4+). The presented results did not support the manufacturer's claim for CK and GGT. Establishing HI thresholds for reporting or suppressing test results is the responsibility of each laboratory, taking into account the manufacturer's data, but also its own investigations.

Effects of recreational SCUBA diving practiced once a week on neurohormonal response and myokines-mediated communication between muscles and the brain.

Objective: During physical activity, activation of muscular, endocrine, and nervous systems, results in intensive crosstalk between muscles and other organs, which enables response to physiological stress. In SCUBA diving, extreme environmental conditions represent an additional challenge for homeostasis maintenance, but underlying mechanisms are largely unknown. We aimed to contribute to the understanding of neurohormonal response and muscle-brain crosstalk by measuring the concentrations of the selected hormones secreted by the pituitary-target organ axis and myokines involved in the muscle-brain endocrine loop in recreational SCUBA (rSCUBA) divers. Methods: Fourteen male divers performed five open-water recreational dives (one per week, depth of 20-30 m, lasting 30 min, between 9 and 10 am), after a winter non-diving period of 5 months. Blood samples were collected immediately before and after the first, third, and fifth dives. Adrenocorticotropic hormone (ACTH), cortisol, thyroid-stimulating hormone (TSH), free thyroxine (fT4), prolactin, total testosterone, growth hormone (GH), insulin-like growth factor-1 (IGF-1), irisin, brain-derived neurotrophic factor (BDNF), S100B, glial fibrillary acidic protein (GFAP), and neuron-specific enolase (NSE) were measured using commercially available immunoassays. Results: Cortisol and ACTH levels decreased after every dive, while total testosterone decreased only after the first dive. No significant changes in post-dive values, as well as the cumulative effect on any other measured hormone, were observed. Although irisin and BDNF levels decreased after the first and third dives, the fifth dive caused a significant increase in both myokines. Changes in IGF-1 levels were not observed. All three dives caused a significant increase in S100B levels. A statistically significant decrease in GFAP concentration was observed after every dive, while NSE pre-dive concentration declined over the studied period. The cumulative effect on myokine levels was reflected in a continuous decline in irisin and BDNF pre-dive levels throughout the studied period, but an increasing trend after the fifth dive was observed. Conclusions: Observed changes in myokines and hormone levels point to a specific response to rSCUBA practiced once a week, most likely due to extreme environmental conditions. Further studies on communication between muscles and other organ systems, particularly on the muscle-brain endocrine loop, are required for a deeper understanding of the adaptation mechanisms to this kind of physiological stress.

Changes in Specific Biomarkers Indicate Cardiac Adaptive and Anti-inflammatory Response of Repeated Recreational SCUBA Diving.

Objective: Recreational SCUBA (rSCUBA) diving has become a highly popular and widespread sport. Yet, information on molecular events underlying (patho)physiological events that follow exposure to the specific environmental conditions (hyperbaric conditions, coldness, immersion, and elevated breathing pressure), in which rSCUBA diving is performed, remain largely unknown. Our previous study suggested that repeated rSCUBA diving triggers an adaptive response of cardiovascular and immune system. To elucidate further molecular events underlying cardiac and immune system adaptation and to exclude possible adverse effects we measured blood levels of specific cardiac and inflammation markers. Methods: This longitudinal intervention study included fourteen recreational divers who performed five dives, one per week, on the depth 20-30 m that lasted 30 min, after the non-dive period of 5 months. Blood samples were taken immediately before and after the first, third, and fifth dives. Copeptin, immunoglobulins A, G and M, complement components C3 and C4, and differential blood count parameters, including neutrophil-to-lymphocyte ratio (NLR) were determined using standard laboratory methods. Cell-free DNA was measured by qPCR analysis and N-glycans released from IgG and total plasma proteins (TPP), were analyzed by hydrophilic interaction ultra-performance liquid chromatography. Results: Copeptin level increased after the first dive but decreased after the third and fifth dive. Increases in immunoglobulins level after every dive and during whole studied period were observed, but no changes in C3, C4, and cfDNA level were detected. NLR increased only after the first dive. IgG and TPP N-glycosylation alterations toward anti-inflammatory status over whole studied period were manifested as an increase in monogalyctosylated and core-fucosylated IgG N-glycans and decrease in agalactosylated TPP N-glycans. Conclusion: rSCUBA diving practiced on a regular basis promotes anti-inflammatory status thus contributing cardioprotection and conferring multiple health benefits.

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.

Time-dependent variation of ionized calcium in serum samples.

INTRODUCTION: The aim of this study was to compare ionized calcium (iCa) concentrations in arterial heparinized blood and venous serum and to investigate time-dependent variation of iCa in serum samples centrifuged and analysed at different times. MATERIALS AND METHODS: Ionized calcium was measured (N = 25) in arterial blood within 20 min after puncture, and in serum within 10 min after centrifugation conducted 30 min after sampling. Effect of time between sampling and centrifugation was examined in three tubes (N = 30) centrifuged 15, 30 and 60 min after sampling, and analysed within 10 min. Effect of time between centrifugation and analysis was investigated in three tubes (N = 31) centrifuged 30 min after sampling and analysed: 0-10, 30-40 and 90-100 min after centrifugation. Ionized calcium was measured on the Siemens RapidLab 348EX analysers. Statistical significance was tested using Wilcoxon test and ANOVA analysis. Clinical significance was judged against reference change values (RCV). RESULTS: No statistically significant difference was found between iCa in arterial blood and serum (P = 0.274). A statistically significant decrease was found: in tubes centrifuged 60 and 15 min after sampling versus 30 min (P = 0.005, P = 0.003); and in tubes analysed 30-40 and 90-100 min after centrifugation versus 0-10 min (P = 0.021, P = 0.027). Clinically significant changes were observed: 60 versus 30 min (centrifugation) and 90-100 versus 0-10 and 30-40 min (analysis). CONCLUSIONS: Timely analysed arterial blood and serum samples can be used interchangeably. To avoid clinically significant variations, serum tubes should be centrifuged within 30 min after sampling, and analysis should be performed within 30 min after centrifugation.

Effect of scuba diving on the oxidant/antioxidant status, SIRT1 and SIRT3 expression in recreational divers after a winter nondive period.

The aim of this study was to examine the effects of scuba diving on oxidative damage markers in erythrocytes and plasma, antioxidant system in peripheral blood mononuclear cells (PBMCs), as well as sirtuin 1 (SIRT1) and sirtuin 3 (SIRT3) gene expressions in recreational divers after a winter nondive period (at least 5 months). For that purpose, 17 male recreational divers performed an immersion at a depth of 30 m for 30 min. Blood samples were collected immediately before and after diving, 3 and 6 h after diving. Erythrocyte lipid peroxidation measured by thiobarbituric-reactive substances (TBARS) method was significantly increased immediately after diving, but returned to the baseline 6 h after diving, while no significant change was found for plasma TBARS and protein carbonyl derivates in both plasma and erythrocytes. Diving-induced catalase (CAT), superoxide dismutase 2 (SOD2), and consequently total superoxide dismutase (SOD) activities in the PBMC samples (significantly increased immediately after diving, reached the maximum activities 3 h after diving, while 6 h after diving only CAT activity remained significantly increased). No significant change was observed for SOD1 activity and gene expression, as well as SOD2 expression, while CAT and SIRT1 expressions were slightly decreased immediately after diving and 3 h after diving. Interestingly, SIRT3 expression was significantly increased 6 h after diving. In conclusion, after the first dive to 30 m after a nondive season, activation of antioxidant defence was not sufficient to prevent oxidative damage, while SIRT3 upregulation could be a step towards an adaptive response to the diving.

Does recreational scuba diving have clinically significant effect on routine haematological parameters?

INTRODUCTION: Scuba diving represents a combination of exercise and changes in environmental conditions. This study aimed to evaluate changes in haematological parameters after recreational scuba diving in order to identify clinically significant changes. MATERIALS AND METHODS: The study included males, 17 recreational divers, median age (range) 41 (30-52) years. Blood samples were taken before diving, immediately after diving to 30 meters for 30 minutes, 3 hours and 6 hours after diving. Complete blood counts were analyzed on the Cell Dyn Ruby haematology analyzer. Statistical significance between successive measurements was tested using Friedman test. The difference between the two measurements was judged against desirable bias (DSB) derived from biological variation and calculated reference change values (RCV). The difference higher than RCV was considered clinically significant. RESULTS: A statistically significant increase and difference judging against DSB was observed: for neutrophils immediately, 3 and 6 hours after diving (18%, 34% and 36%, respectively), for white blood cells (WBCs) 3 and 6 hours after diving (20% and 25%, respectively), for lymphocytes (20%) and monocytes (23%) 6 hours after diving. A statistically significant decrease and difference judging against DSB was found: immediately after diving for monocytes (- 15%), 3 and 6 hours after diving for red blood cells (RBCs) (- 2.6% and -2.9%, respectively), haemoglobin (- 2.1% and - 2.8%, respectively) and haematocrit (- 2.4% and - 3.2%, respectively). A clinically significant change was not found for any of the test parameters when compared to RCV. CONCLUSIONS: Observed statistically significant changes after recreational scuba diving; WBCs, neutrophils, lymphocytes, monocytes increase and RBCs, haemoglobin, haematocrit decrease, probably will not affect clinical decision.

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.