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Background: The aim of this work was to investigate the serum amino acid (AA) changes after a breath-hold diving (BH-diving) training session under several aspects including energy need, fatigue tolerance, nitric oxide (NO) production, antioxidant synthesis and hypoxia adaptation. Twelve trained BH-divers were investigated during an open sea training session and sampled for blood 30 min before the training session, 30 min and 4 h after the training session. Serum samples were assayed for AA changes related to energy request (alanine, histidine, isoleucine, leucine, lysine, methionine, proline threonine, valine), fatigue tolerance (ornithine, phenylalanine, tyrosine), nitric oxide production (citrulline), antioxidant synthesis (cystine, glutamate, glycine) and hypoxia adaptation (serine, taurine)., Main Results: Concerning the AA used as an energy support during physical effort, we found statistically significant decreases for all the investigated AA at T1 and a gradual return to the basal value at T2 even if alanine, proline and theonine still showed a slight significant reduction at this time. Also, the changes related to the AA involved in tolerance to physical effort showed a statistically significant decrease only at T1 respect to pre-diving value and a returned to normal value at T2. Citrulline, involved in NO production, showed a clear significant reduction both at T1 and T2. Concerning AA involved in endogenous antioxidant synthesis, the behaviour of the three AA investigated is different: we found a statistically significant increase in cystine both at T1 and T2, while glycine showed a statistically significant reduction (T1 and T2). Glutamate did not show any statistical difference. Finally, we found a statistically significant decrease in the AA investigated in other hypoxia conditions serine and taurine (T1 and T2)., Conclusions: Our data seem to indicate that the energetic metabolic request is in large part supported by AA used as substrate for fuel metabolism and that also fatigue tolerance, NO production and antioxidant synthesis are supported by AA. Finally, there are interesting data related to the hypoxia stimulus that indirectly may confirm that the muscle apparatus works under strong exposure conditions notwithstanding the very short/low intensity of exercise, due to the intermittent hypoxia caused by repetitive diving., (© 2022. The Author(s).)

Background: Breath-hold diving (BH-diving) is associated to extreme environmental conditions, prolonged physical activity, and complex adaptation mechanisms to supply enough O 2 to vital organs. Consequently, one of the biggest effects could be an increased exercise-induced muscle fatigue, in both skeletal and cardiac muscles that can induce an increase of muscles injury markers including creatine kinase (CK), aspartate transferase (AST), and alanine transferase (ALT) when concerning the skeletal muscle, cardiac creatine kinase isoenzyme (CK-MBm) and cardiac troponin I (cTnI) when concerning the cardiac muscle, and lactate dehydrogenase (LDH) as index of muscle stress. The aim of this study is to investigate serum cardiac and skeletal muscle markers before and after a BH-diving training session., Results: We found statistically significant increases of CK (T0: 136.1% p < 0.0001; T1: 138.5%, p < 0.0001), CK-MBm (T0: 145.1%, p < 0.0001; T1: 153.2%, p < 0.0001) LDH (T0: 110.4%, p < 0.0003; T1: 110.1%, p < 0.0013) in both T0 and T1 blood samples, as compared to basal value. AST showed a statistically significant increase only at T0 (106.8%, p < 0.0007) while ALT did not exhibit statistically significant changes. We did not find any changes in cTnI levels between pre-dive and post-dive samples., Conclusions: Our data seem to indicate that during a BH-diving training session, skeletal and cardiac muscles react to physical effort releasing stress-related substances. Although the peculiar nature of BH-diving makes it difficult to understand if our results are related only to exercise induced muscle adaptation or whether acute hypoxia or a response to environmental changes (pressure) play a role to explain the observed changes, further studies are needed to better understand if these biomarker changes are linked to physical exercise or to acute hypoxia, or if both conditions play a role., (© 2021. The Author(s).)

Introduction: Nitric oxide (NO) is an essential signaling molecule modulating the endothelial adaptation during breath-hold diving (BH-diving). This study aimed to investigate changes in NO derivatives (NOx) and total antioxidant capacity (TAC), searching for correlations with different environmental and hyperbaric exposure., Materials and Methods: Blood samples were obtained from 50 breath-hold divers (BH-divers) before, and 30 and 60 min after the end of training sessions performed both in a swimming pool or the sea. Samples were tested for NOx and TAC differences in different groups related to their hyperbaric exposure, experience, and additional genetic polymorphism., Results: We found statistically significant differences in NOx plasma concentration during the follow-up (decrease at T30 and increase at T60) compared with the pre-dive values. At T30, we found a significantly lower decrease of NOx in subjects with a higher diving experience, but no difference was detected between the swimming pool and Sea. No significant difference was found in TAC levels, as well as between NOx and TAC levels and the genetic variants., Conclusion: These data showed how NO consumption in BH-diving is significantly lower in the expert group, indicating a possible training-related adaptation process. Data confirm a significant NO use during BH-diving, compatible with the well-known BH-diving related circulatory adaptation suggesting that the reduction in NOx 30 min after diving can be ascribed to the lower NO availability in the first few minutes after the dives. Expert BH-divers suffered higher oxidative stress. A preliminary genetic investigation seems to indicate a less significant influence of genetic predisposition., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Cialoni, Brizzolari, Samaja, Bosco, Paganini, Sponsiello, Lancellotti and Marroni.)

Background: The physiological and pathophysiological mechanisms that govern diving, both self-contained underwater breathing apparatus (SCUBA) and breath-hold diving (BH-diving), are in large part well known, even if there are still many unknown aspects, in particular about cell metabolism during BH-diving. The scope of this study was to investigate changes in glycemia, insulinemia, and the catecholamine response to BH-diving, to better understand if the insulin-stimulated glucose uptake mechanism is involved in cellular metabolism in this sport., Methods: Twenty male experienced healthy breath-hold divers were studied. Anthropometric information was obtained. Glycemia, insulinemia, and catecholamine response were investigated before and after the series of BH-diving., Results: We found a statistically significant decrease in the blood glucose levels between before and after dives (mean 94.3 ± 11.6 vs. 83.5 ± 12.5 mg/dl) P = 0.001 and a statistically significant increase in blood insulin value (median 4.5 range 3.4/6.4 vs. 7.0 range 4.2/10.2 mcgU/ml) P < 0.0001. Also, we found a statistically significant increase of catecholamine production (median 14.0 range 8/18 vs. 15.5 range 10.0/21.0 μg) P < 0.0001., Conclusions: The increase in blood insulin during BH-diving associated with the decrease of blood glucose levels could indicate that the upregulating cellular uptake is not caused by activation of the specific glucose transporters. Particular diving-related conditions such as the diving reflex, the intermittent hypoxia/hyperoxia, and the particular environmental condition could play an important role in the mechanism involved in glycemia decrease in BH-diving. Our data confirm that the adaptations to BH-diving are caused by complex mechanisms and involve many peculiar responses still in large part unknown.

Introduction: Breath-hold diving-induced hemoptysis (BH-DIH) has been reported in about 25% breath-hold divers (BHD) and is characterized by dyspnea, coughing, hemoptysis and chest pain. We investigated whether eNOS G894T, eNOS T786C and ACE insertion/deletion I/D genetic variants, are possible BH-DIH risk factors., Methods: 108 experienced healthy instructor BHDs with the same minimum requirements (102 male, six female; mean age 43.90 ± 7.49) were studied. We looked for different eNOS G894T, eNOS T786C and ACE insertion/ deletion genetic variants between BH-DIH-positive and BH-DIH-negative subjects to identify the variants most frequently associated with BH-DIH., Results: At least one BH-DIH episode was reported by 22.2% of subjects, while 77.7% never reported BH-DIH. The majority of BH-DIH-positive subjects showed eNOS G894T (p = 0.001) and eNOS-T786C (p = 0.001) genotype "TT" (high-risk profile). Prevalence of BH-DIH was higher in subjects with eNOS G894T TT genotype (50%) than in subjects with GT (9.5%, p < 0.001) and GG (24%, (p = 0.0002) genotype (low-risk profile). Similar results were observed for eNOS T786C: BH-DIH prevalence was higher in the TT genotype (41.2%) group than in the CT (15.4%, p < 0.001) and CC genotype (9.1%, p < 0.001) groups. BH-DIH prevalence was significantly higher in subjects showing ACE ID genotype (34.5%) than II (0%, p < 0.001) and DD (10.5%, p = 0.0002). Of the ACE "II" genotype group, 100% never developed BH-DIH., Discussion: eNOS-G894T, eNOS-T786C and ACE influence NO availability and regulation of peripheral vascular tone and blood flow. Different genetic variants of eNOS-G894T, eNOS-T786C and ACE appear significantly related to the probability to develop BH-DIH (p < 0.001).

Introduction: Hyperoxia causes oxidative stress. Breath-hold diving is associated with transient hyperoxia followed by hypoxia and a build-up of carbon dioxide (CO₂), chest-wall compression and significant haemodynamic changes. This study analyses variations in plasma oxidative stress markers after a series of repetitive breath-hold dives., Methods: Thirteen breath-hold divers were asked to perform repetitive breath-hold dives to 20 metres' depth to a cumulative breath-hold time of approximately 20 minutes over an hour in the open sea. Plasma nitric oxide (NO), peroxinitrites (ONOO⁻) and thiols (R-SH) were measured before and after the dive sequence., Results: Circulating NO significantly increased after successive breath-hold dives (169.1 ± 58.26% of pre-dive values; P = 0.0002). Peroxinitrites doubled after the dives (207.2 ± 78.31% of pre-dive values; P = 0.0012). Thiols were significantly reduced (69.88 ± 19.23% of pre-dive values; P = 0.0002)., Conclusion: NO may be produced by physical effort during breath-hold diving. Physical exercise, the transient hyperoxia followed by hypoxia and CO₂ accumulation would all contribute to the increased levels of superoxide anions (O₂²⁻). Since interaction of O₂²⁻ with NO forms ONOO⁻, this reaction is favoured and the production of thiol groups is reduced. Oxidative stress is, thus, present in breath-hold diving.

Objective: Scuba and breath-hold divers are compared to investigate whether endothelial response changes are similar despite different exposure(s) to hyperoxia., Design: 14 divers (nine scuba and five breath-holding) performed either one scuba dive (25m/25 minutes) or successive breath-hold dives at a depth of 20 meters, adding up to 25 minutes of immersion time in a diving pool. Flow-mediated dilation (FMD) was measured using echography. Peripheral post-occlusion reactive hyperemia (PORH) was assessed by digital plethysmography and plasmatic nitric oxide (NO) concentration using a nitrate/nitrite colorimetric assay kit., Results: The FMD decreased in both groups. PORH was reduced in scuba divers but increased in breath-hold divers. No difference in circulating NO was observed for the scuba group. Opposingly, an increase in circulating NO was observed for the breath-hold group., Conclusion: Some cardiovascular effects can be explained by interaction between NO and superoxide anion during both types of diving ending to less NO availability and reducing FMD. The increased circulating NO in the breath-hold group can be caused by physical exercise. The opposite effects found between FMD and PORH in the breath-hold group can be assimilated to a greater responsiveness to circulating NO in small arteries than in large arteries.

Introduction: After repetitive deep dives, breath-hold divers are often affected by a syndrome characterized by typical symptoms such as cough, sensation of chest constriction, blood-striated expectorate (hemoptysis) and, rarely, an overt acute pulmonary edema syndrome, often together with various degrees of dyspnea. The aim of this work is an epidemiological investigation to evaluate the prevalence of acute respiratory symptoms (ARS) in breath-hold divers (BHDs) in practicing breath-hold diving., Materials and Methods: A retrospective investigation has been performed using specific questionnaires completed by a selected sample of free-divers (212 breath-hold diving instructors--194 male, 18 female; mean age 34 +/- 6.91 years); affiliated with Apnea Academy, (International School for Education and Research of Free-Diving). We also investigated possible risk factors for post-dive acute respiratory symptoms. Furthermore, the authors report that a severe case of acute pulmonary edema occurred to a healthy and experienced breath-hold diving instructor. We reported detailed CT scan and follow-up CT scans three days later, with another scan reported 10 days later as well., Result: A total of 56 subjects (26.4%) reported previous events such as cough, thoracic constraint, hemoptysis, associated with various degrees of dyspnea as confirmation of pulmonary involvement. Forty-five of them (82%) reported signs of true hemoptysis and a high degrees of dyspnea. A CT scan revealed the presence of patchy bilateral lung opacities at the level of superior and parahilar zones; follow-up CT scans three days later and 10 days later are also reported., Conclusion: Our data show that this is a common condition among experienced BHDs. In our opinion, this is particularly interesting for the free-diving community.

The purpose of the study was to analyze the ultrasound lung comets (ULCs) variation, which are a sign of extra-vascular lung water. Forty-two healthy individuals performed breath-hold diving in different conditions: dynamic surface apnea; deep variable-weight apnea and shallow, face immersed without effort (static maximal and non-maximal). The number of ULCs was evaluated by means of an ultrasound scan of the chest, before and after breath-hold diving sessions. The ULC score increased significantly from baseline after dynamic surface apnea (p = 0.0068), after deep breath-hold sessions (p = 0.0018), and after static maximal apnea (p = 0.031). There was no statistically significant difference between the average increase of ULC scores after dynamic surface apnea and deep breath-hold diving. We, therefore, postulate that extravascular lung water accumulation may be due to other factors than (deep) immersion alone, because it occurs during dynamic surface apnea as well. Three mechanisms may be responsible for this. First, the immersion-induced hydrostatic pressure gradient applied on the body causes a shift of peripheral venous blood towards the thorax. Second, the blood pooling effect found during the diving response Redistributes blood to the pulmonary vascular bed. Third, it is possible that the intense involuntary diaphragmatic contractions occurring during the "struggle phase" of the breath-hold can also produce a blood shift from the pulmonary capillaries to the pulmonary alveoli. A combination of these factors may explain the observed increase in ULC scores in deep, shallow maximal and shallow dynamic apneas, whereas shallow non-maximal apneas seem to be not "ULC provoking".

To define the dynamics of cardiovascular adjustments to apnoea during immersion, beat-to-beat heart rate (HR) and systolic (SBP) and diastolic (DBP) blood pressures were recorded in six divers during and after prolonged apnoeas while resting fully immersed in 27 degrees C water. Apnoeas lasted 215 +/- 35 s. Compared to control values, HR decreased by 20 beats min(-1) and SBP and DBP increased by 23 and 17 mmHg, respectively, in the initial 20 +/- 3 s (phase I). Both HR and BP remained stable during the following 92 +/- 15 s (phase II). Subsequently, during the final 103 +/- 29 s, SBP and DBP increased linearly to values about 60% higher than control, whereas HR remained unchanged (phase III). Cardiac output (Q') decreased by 35% in phase I and did not further change in phases II and III. Compared to control, total peripheral resistances were twice and three times higher than control, respectively, at the end of phases I and III. After resumption of breathing, HR and BP returned to control values in 5 and 30 s, respectively. The time courses of cardiovascular adjustments to immersed breath-holding indicated that cardiac response took place only at the beginning of apnoea. In contrast, vascular responses showed two distinct adjustments. This pattern suggests that the chronotropic control via the baroreflex is modified during apnoea. These cardiovascular changes during immersed static apnoea are in agreement with those already reported for static dry apnoeas.