Extracellular Vesicles in Environmental Toxicological Studies: Association between Urinary Concentrations of Phthalate Metabolites and Exosomal miRNA Expression Profiles.

Phthalates are chemical compounds, mainly used as additives in plastics, which are known to induce harmful impacts to the environment and human health due to their ability to act as hormone-mimics. Few studies have been reported on the relationship between human exposure to phthalates and the level of circulating microRNAs (miRs), especially those miRs encapsulated in extracellular vesicles/exosomes or exosome-like vesicles (ELVs). We examined the relationship of ELV-miR expression patterns and urine of adult men with five phthalate metabolites (i.e., mono isobutyl phthalate, mono-n-butyl phthalate, mono benzyl phthalate, mono-(2-ethyl-5-oxohexyl) phthalate, mono-(2-ethylhexyl) phthalate) to identify potential biomarkers and relevant pathways. We found significant positive associations which were further confirmed by multivariable analysis. Overall, our analyses showed that the Σ phthalate metabolite concentration was associated with a significant increase in the expression level of two miRs found in ELV: miR-202 and miR-543. Different pathways including cancer and immune-related responses were predicted to be involved in this relationship. Analyzing the specific downstream target genes of miR-202 and miR-543, we identified the phosphatase and tensin homolog (PTEN) as the key gene in several converging pathways. In summary, the obtained results demonstrate that exposure to environmental phthalates could be related to altered expression profiles of specific ELV-miRs in adult men, thereby demonstrating the potential of miRs carried by exosomes to act as early effect biomarkers.

Endurance-dependent urinary extracellular vesicle signature: shape, metabolic miRNAs, and purine content distinguish triathletes from inactive people.

Extracellular vesicles (EVs) enriched with bioactive molecules have gained considerable attention in nanotechnology because they are critical to intercellular communication while maintaining low immunological impact. Among biological matrices, urine has emerged as a noninvasive source of extracellular-contained liquid biopsy, currently of interest as a readout for physiological adaptations. Therefore, we aimed to evaluate chronic adaptations of endurance sport practice in terms of urinary EV parameters and evaluated by food consumption assessment. Two balanced groups of 13 inactive controls vs. triathlon athletes were enrolled; their urinary EVs were obtained by differential ultracentrifugation and analyzed by dynamic light scattering and transmission electron and atomic force microscopy. The cargo was analyzed by means of purine and miRNA content through HPLC-UV and qRT-PCR. Specific urinary EV signatures differentiated inactive versus endurance-trained in terms of peculiar shape. Particularly, a spheroid shape, smaller size, and lower roughness characterize EVs from triathletes. Metabolic and regulatory miRNAs often associated with skeletal muscle (i.e., miR378a-5p, miR27a-3p, miR133a, and miR206) also accounted for a differential signature. These miRNAs and guanosine in urinary EVs can be used as a readout for metabolic status along with the shape and roughness of EVs, novel informative parameters that are rarely considered. The network models allow scholars to entangle nutritional and exercise factors related to EVs' miRNA and purine content to depict metabolic signatures. All in all, multiplex biophysical and molecular analyses of urinary EVs may serve as promising prospects for research in exercise physiology.

Neurovegetative and Emotional Modulation Induced by Mozart's Music.

INTRODUCTION: Since decades, the "Mozart effect" has been studied. However, the diverse effects of Mozart's music components have not been yet defined. Authors aimed to identify a differential response to short-term exposure to Mozart's music, or to its rhythmic signature only, on subjective and objective measures. METHODS: The Mozart Sonata in A major K 331 (Mozart), the same piece consisting only of beat (Destructured), and duration-matched silence were administered to 25 healthy young adults, stood supine in a relaxing setting. The Italian Mood Scale questionnaire was administered before and after each listening. Heart rate variability (HRV) metrics were calculated from ECG recording, and breath flow was registered during experiments. RESULTS: After Destructured, there was no change of fatigue and tension. After Mozart, fatigue was significantly reduced (and a tendency appeared for tension), whereas vigor was not. Breathing rate tended to be higher during Mozart. The nonlinear parameter HFD of HRV analysis, even though not significantly, was slightly lower during Destructured; Poincaré plots SD1 and SD2 tended to be lower during Mozart. DISCUSSION/CONCLUSION: Mozart's music may allow to maintain arousal during relaxing condition. Psychological response of music and physiological dynamics were not necessarily entangled. Musical pieces based on individual physiological signature may lead musical psychological interventions.

Olfactory Response to Altitude Hypoxia: A Pilot Study During a Himalayan Trek.

The adaptation of olfaction to extreme environments is an area of limited understanding. This study aimed to get insights into the factors that constrain olfactory function at high altitudes. To this end, we compared the results of smell tests performed on the same subjects at low (665 m) and high altitude (4,780 m) during the "Kanchenjunga Exploration and Physiology" project in the Himalayas. The most distinct adaptive differences found at high altitude were reductions in the odor intensity, color-odor bimodal association, and memory, and increases in flavor perception. Physiological adaptations of the sense of smell resulting from exposure to hypoxia at altitude may have explanatory validity in unraveling the mechanisms that shape changes in olfactory function in the processes of aging and neurodegenerative diseases.

Fertility Impairment after Trekking at High Altitude: A Proof of Mechanisms on Redox and Metabolic Seminal Changes.

Many authors described negative but reversible effects of high-altitude hypoxic exposure on animal and human fertility in terms of sperm concentration, function, and biochemical alterations. The aim of this study was to evaluate the acute and chronic effects of high-altitude exposure on classical sperm parameters, redox status, and membrane composition in a group of travellers. Five healthy Italian males, all lowlanders not accustomed to the altitude, were evaluated after 19 days-trekking through low, moderate, and high altitudes in the Himalayas. Sperm samples were collected before (Pre), 10 days after (Post), and 70 days after the end of the expedition (Follow-up). Sperm concentration, cholesterol and oxysterol membrane content, and redox status were measured. Hypoxic trek led to a significant reduction in sperm concentration (p < 0.001, η2p = 0.91), with a reduction from Pre to Post (71.33 ± 38.81 to 60.65 ± 34.63 × 106/mL) and a further reduction at Follow-up (to 37.13 ± 39.17 × 106/mL). The seminal volume was significantly affected by the hypoxic trek (p = 0.001, η2p = 0.75) with a significant reduction from Pre to Post (2.86 ± 0.75 to 1.68 ± 0.49 mL) and with partial recovery at Follow-up (to 2.46 ± 0.45 mL). Moreover, subjects had an increase in ROS production (+86%), and a decrease in antioxidant capacity (−37%) in the Post period with partial recovery at Follow-up. These results integrated the hormonal response on thyroid function, hypothalamus−pituitary−gonadal axis, and the prolactin/cortisol pathways previously reported. An uncontrolled ROS production, rather than a compromised antioxidant activity, was likely the cause of impaired sperm quality. The reduction in fertility status observed in this study may lie in an evolutionary Darwinian explanation, i.e., limiting reproduction due to the "adaptive disadvantage" offered by the combined stressors of high-altitude hypoxia and daily physical exercise.

Pathophysiological Responses to a Record-Breaking Multi-hour Underwater Endurance Performance: A Case Study.

The "Endless Diving Project-Step 36" took place in the harbor waters of the town of Maratea in Italy in September 2014. The goal of the project was an attempt by an experienced male diver, equipped with a wet 7-mm suit and a normal gas tank, to set the world record-breaking of nonstop underwater performance. We studied inflammatory, hematological, and endocrine responses during the extreme condition of the attempt. Venous blood samples were collected at baseline, the day before the attempt; immediately after the return from underwater; then at Day 1, Day 4, and Day 12; and later at Month 1 and Month 41 of follow-up. We found that there was an increase in the content of blood neutrophils, monocytes, and eosinophils and a decrease in lymphocytes at Day 1 and a late increase in basophils at Day 12 after the dive. Inflammatory markers and hematocrit and hemoglobin increased immediately after the dive, dropped at Day 1, and reverted gradually to the control level from Day 4 to Day 12. Serotonin and dopamine decreased, while adrenaline increased at Day 1, gradually recovering in the days of follow-up. Insulin, luteinizing hormone, growth hormone, and prolactin increased, while testosterone, cortisol, 17ß-estradiol, thyroid-stimulating hormone, and adrenocorticotropic hormone decreased at Day 1, with a partial recovery at Day 4. We conclude that the homeostatic response to the extreme, prolonged underwater performance showed signs of psychological and pro-inflammatory stress. The hormonal response reflected an acute testicular insufficiency. These responses resembled those characteristics for ultra-endurance exercise accompanied by vasculitis and dehydration.

Effects of Physical Activity at High Altitude on Hormonal Profiles in Foreign Trekkers and Indigenous Nepalese Porters.

Altitude exposure affects hormonal homeostasis, but the adaptation of different populations is still not finely defined. This study aims to compare the mid-term effects of combining physical activity and altitude hypoxia on hormonal profiles in foreign trekkers coming from Italy versus indigenous Nepalese porters during a Himalayan trek. Participants (6 Italians and 6 Nepalese) completed a 300 km distance in 19 days of an accumulated altitude difference of 16,000 m, with an average daily walk of 6 h. The effect of high altitude on hormonal pathways was assessed by collecting blood samples the day before the expedition and the day after its completion. Foreign trekkers had an additional follow-up sample collected after 10 days. The findings revealed a different adaptation of thyroidal and gonadal axes to mid-term strenuous physical activity combined with high-altitude hypobaric hypoxia. The thyroid function shifted to the protective mechanism of low free triiodothyronine (FT3), whereas the gonadal axis was suppressed. The Italian trekkers and Nepalese porters had lower total testosterone and 17-ß-estradiol levels after the expedition. At the follow-up, the Italians had increased testosterone values. Prolactin secretion decreased in the Italians but increased in the Nepalese. We conclude that exposure to high-altitude affects the hormonal axes. The effect seems notably pronounced for the hypothalamus-pituitary gonadal axis, suppressed after high-altitude exposure.

Uroflowmetry and Altitude Hypoxia: A Report from Healthy Italian Trekkers and Nepali Porters During Himalayan Expedition.

Hypoxia alters micturition, which influences bladder function by involving different neurological and humoral systems. In this study we assessed the mid-term effects of altitude hypoxia on uroflowmetry in healthy male lowlander native Nepali porters and Italian trekkers, four each, who coattended a Himalayan expedition. All the participants completed a 19-day trek along a demanding route with ascent and descent at the Kanchenjunga Mountain. They underwent micturition and urodynamic analysis twice, at low altitude of 665 m a.s.l. and high altitude of 4,750 m a.s.l. Statistical comparisons considered the altitude effects (low vs. high) and ethnicity (Italian vs. Nepali). Food consumption was recorded, and water and energy intake were calculated. We found trends of borderline significance in the mean urinary flow rate (Qmean) (p = 0.058; effect size η2 p = 0.478) and in Qmax to the advantage of the Nepali. There was no evidence of differences when comparing time to Qmax and urine volume at Qmax and Qmean for altitude or altitude × ethnicity. In addition, there was a lonely female participant, who, analyzed as a case report, showed increased Qmean at high altitude. Older age mitigated while energy intake potentiated the ethnic differences noted in uroflowmetry. We conclude that altitude hypoxia rather inappreciably affects micturition in healthy men. However, a trend for possible ethnic differences raises worthy of note perspectives on adaptive ability of micturition. Also, dietary intake and age should be considered as confounding elements when evaluating micturition.

Urinary physiology and hypoxia: a pilot study of moderate-altitude trekking effects on urodynamic indexes.

Exposure to high altitude is one of the most widely used models to study the adaptive response to hypoxia in humans. However, little is known about the related effects on micturition. The present study addresses the adaptive urinary responses in four healthy adult lowlanders, comparing urodynamic indexes at Kathmandu [1,450 m above sea level (a.s.l.); K1450] and during a sojourn in Namche Bazar (3,500 m a.s.l.; NB3500). The urodynamic testing consisted of cistomanometry and bladder pressure/flow measurements. Anthropometrics, electrocardiographic, and peripheral capillary oxygen saturation data were also collected. The main findings consisted of significant reductions in bladder power at maximum urine flow by ~30%, bladder contractility index by 13%, and infused volume both at first (by 57%) and urgency sensation (by 14%) to urinate, indicating a reduced cystometric capacity, at NB3500. In addition to the urinary changes, we found that oxygen saturation, body mass index, body surface area, and median RR time were all significantly reduced at altitude. We submit that the hypoxia-related parasympathetic inhibition could be the underlying mechanism of both urodynamic and heart rate adaptive responses to high-altitude exposure. Moreover, increased diuresis and faster bladder filling at altitude may trigger the anticipation of being able to void, a common cause of urgency. We believe that the present pilot study represents an original approach to the study of urinary physiology at altitude.

Physiological and pathological levels of prostaglandin E2 in renal parenchyma and neoplastic renal tissue.

Prostaglandin (PG)E2 seems to promote tumor proliferation by regulating cell growth, inhibiting apoptosis, promoting angiogenesis, and suppressing host immune surveillance of cancer cells. The suppression of prostaglandins biosynthesis is thought to be the main molecular mechanism for non-steroidal anti-inflammatory drugs antineoplastic effect. Yet the relationship between PGE2 and human renal cell carcinoma remains unclear. The aim of our study is to evaluate the PGE2 content in human renal parenchyma and Renal Cell Carcinoma. The study was conducted on 20 consecutive patients undergoing radical nephrectomy for Renal Cell Carcinoma. In the normal renal parenchyma and in the neoplastic renal tissue the PGE2 level was 83.43 ± 5.89 pg/mg and 289.67 ± 22.2 pg/mg, respectively (P < 0.0001). There was no relationship between PGE2 content and Renal Cell Carcinoma dimension, Fuhrman grade, pathological-Tumor-Node and Metastasis (pTNM) stage and histological subtype. The PGE2 over-content in neoplastic renal tissue suggests a role of PGE2 in development and progression of renal carcinoma.

Sex Hormones Response to Physical Hyperoxic and Hyperbaric Stress in Male Scuba Divers: A Pilot Study.

The use of hyperbaric oxygen plays a significant role in many aspects of medicine. However, there are few studies that analyzed the role of hyperbaric oxygen, in addition to physical exercise, on the endocrine profile. The aim of this study was to compare changes in plasma male sex hormones after hyperbaric physical exercise with different hyperbaric oxygen pre-conditionings. We recruited six healthy, well-trained recreational male divers. Concentrations of prolactin (PRL), follicle-stimulating hormone (FSH), luteotrophic hormone (LH), cortisol, 17-ß estradiol (E2), and total testosterone (TT) were measured in venous blood immediately after four different study conditions. Exercise increased PRL and hyperbaric oxygen potentiated this effect. Hyperbaria stimulated the E2 reduction and hyperoxia partially inhibited this reduction. Hyperbaria, but not hyperoxia, stimulated the TT reduction. There were no changes in FSH, LH, and cortisol. The increase in PRL likely reflects a stress response after physical exercise, amplified by hyperbaric oxygen. TT reduction may be interpreted as an acute and transient fertility impairment. Age, blood pressure, and BMI were taken into account as covariates for statistical analyses, and they significantly affected the results, in particular TT. These data open new insight into the role of E2 and PRL in male endocrine adaptive responses.

Body Composition and Endocrine Adaptations to High-Altitude Trekking in the Himalayas.

Long-term exposure to high altitude causes adaptive changes in several blood biochemical markers along with a marked body mass reduction involving both the lean and fat components. The aim of this study was to evaluate the impact of extended physical strain, due to extensive trekking at high altitude, on body composition, selected biomarkers in the blood, and the protective role of a high-protein diet in muscle dysfunction. We found that physical strain at high altitude caused a significant reduction in body mass and body fat, with a concomitant increase in the cross-sectional area of thigh muscles and an unchanged total lean body mass. Further, we found reductions in plasma leptin and homocysteine, while myoglobin, insulin, and C-reactive protein significantly increased. Creatine kinase, lactate dehydrogenase, and leptin normalized per body fat were unchanged. These findings demonstrate that high-altitude hypoxia, involving extended physical effort, has an impact on muscle function and body composition, facilitating sarcopenia and affecting body mass and fat distribution. It also activates pro-inflammatory metabolic pathways in response to muscular distress. These changes can be mitigated by a provision of a high-protein diet.

The influence of altitude hypoxia on uroflowmetry parameters in women.

There is scientific evidence to suggest a correlation between hypoxia and the physiology of micturition. During a Himalayan Scientific and Mountaineering Expedition, we performed tests to investigate the functional interactions between altitude hypoxia and uroflowmetry parameters in women. The tests were carried out in seven women (36.3 ± 7.1 yr) from normoxic [1,340 meters above sea level (m a.s.l.)] to hypoxic conditions (up to 5,050 m a.s.l.) and during the return descent. The following measures were determined: uroflowmetry parameters and saturation of peripheral oxygen (SpO2 ). As expected, SpO2 decreased from 97.7 to 77.8% with increasing altitude. Micturition flow time, flow volume, and voiding time increased with altitude (P < 0.04 for all), indicating a negative correlation with SpO2 In conclusion, in young adult women, micturition physiological parameters were affected during adaptation to hypoxia; the correlation with SpO2 strongly suggests a role of hypoxia in these changes. These data could help to support the design of new strategies for both prevention and medical treatment. An example of the latter might be hyperbaric oxygen therapy, which in some studies has proved able to reduce the symptoms in patients with hypoxic bladder.

[Steroid-Dependent Nephrotic Syndrome Due to Minimal Change Glomerulonephritis Treated with Rituximab].

47-year-old woman suffering from minimal lesion glomerulonephritis previously undergone high-dose steroid therapy and subjected to exacerbations of nephrotic syndrome after therapy discontinuation. It was decided to initiate off-label treatment with Rituximab at a dosage of 375 mg/m2 administred at zero-time, one-month and three months with good therapeutic response and resolution of the clinical laboratory picture. The therapy was well tolerated and had no side effects. This scheme could be an alternative to the conventional therapeutic scheme with steroids or other classes of immunosuppressive drugs, especially in order to avoid problems related to prolonged exposure to steroid therapy.

Myalgic encephalomyelitis/chronic fatigue syndrome from current evidence to new diagnostic perspectives through skeletal muscle and metabolic disturbances.

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a demanding medical condition for patients and society. It has raised much more public awareness after the COVID-19 pandemic since ME/CFS and long-COVID patients share many clinical symptoms such as debilitating chronic fatigue. However, unlike long COVID, the etiopathology of ME/CFS remains a mystery despite several decades' research. This review moves from pathophysiology of ME/CFS through the compelling evidence and most interesting hypotheses. It focuses on the pathophysiology of skeletal muscle by proposing the hypothesis that skeletal muscle tissue offers novel opportunities for diagnosis and treatment of this syndrome and that new evidence can help resolve the long-standing debate on terminology.

Nutritional physiology and body composition changes during a rapid ascent to high altitude.

Exposure to high altitude might cause the body to adapt with negative energy and fluid balance that compromise body composition and physical performance. In this field study involving 12 healthy adults, sex-balanced, and aged 29 ± 4 years with a body mass index of 21.6 ± 1.8 kg/m2, we investigated the effects of a 4-day trekking up to 4556 m a.s.l. on Monte Rosa (Alps, Italy). The food intake was recorded using food diaries and nutrient averages were calculated. The bio-impedance analysis was performed at low and high altitudes, and a wearable biosensor (Swemax) was used to track hydro-saline losses in two participants. Daily total energy intake was 3348 ± 386 kcal for males and 2804 ± 415 kcal for females (13%-14% protein, 35% fat, 44%-46% carbohydrates). Although there was a significant body weight loss (65.0 ± 9.3 vs. 64.2 ± 9.10 kg, p < 0.001, d = 1.398), no significant changes in body composition parameter were found but a trend in the increase of the bioelectrical phase angle in males (p = 0.059, d = -0.991). Body water percentage significantly changed (p = 0.026, η2 p = 0.440), but the absolute water did not, suggesting that the weight loss was not due to water loss. Salivary and urinary osmolality did not change. A reduction in sweat rate at higher altitudes was observed in both participants. Interestingly, salivary leptin increased (p = 0.014, η2 p = 0.510), and salivary ghrelin decreased (p = 0.036, η2 p  = 0.403). Therefore, the 4-day trekking at altitude of hypoxia exposure induced changes in satiety and appetite hormones. High altitude expeditions require more specific nutritional guidance, and using multiplex analysis could help in monitoring fluid balance and body composition.

Comments on: Electromyographic signature of isometric squat in the highest refuge in Europe.

We read the comment by Sarabon and Sasek (Eur J Transl Myol 11846, 2023 doi: 10.4081/ejtm.2023.11846) on our study on the electromyographic results during squat at high-altitude. Their highlighting of the factors that may have biased our results is commendable, despite they criticized our work as if it were a controlled laboratory study. We considered the limitations and argued our interpretation with caution. We found no changes in median frequency, frequency's slope and conduction velocity of EMG signals at high altitude, and a slight decrease of root mean square. We argued that our results may have been due to a preventative mechanism that compensates for relatively greater effort during a fatiguing strength test involving large muscle masses, or to cumulative exertional stress on the muscles. The authors of the comment themselves somehow agree with these interpretations. Guidelines for collecting, reporting and interpreting data from EMG to obtain original information on the neuromuscular system should be integrated with the effort to maintain as much as possible those field conditions that offer unique opportunities.

Sports performance adaptations through occlusal splint: Case reports of triathlon athletes.

OBJECTIVE: Occlusal disturbances affect human posture and sports performance. This study aimed to monitor biomechanical adaptations to personalized occlusal splints. METHODS: Splints were customized based on stabilometry, thermography, sEMG, and kinesiography, and administered to three triathlon athletes. They were evaluated during a 4-month period, using isokinetic indexes, running kinematics and anaerobic outputs. RESULTS: Individuality emerged as a key factor driving type, quantity, quality, and time trajectories of adaptations. The use of instrumental and clinical tests allowed the detection of static balance and biting function improvements, but not necessarily parallel to sports performance improvements. CONCLUSION: The authors argue that strength and kinematic imbalances are joint and task-specific and support the use of multi-stage monitoring of the biomechanical effect of mouthpieces. Kinematics of cycling and running may be widely assessed with ecological and inexpensive methods. Strength imbalances need to be continuously monitored due to the high informative value to injury prevention.

Investigating Auditory Perception at Europe's Highest Mountain Lodge.

Altitude hypoxia can impair sensory and cognitive functions, from causing slowed responses to sensory hallucinations. In a field study, we tested 12 expeditioners at varied altitudes (low: 1696 m; high: 4556 m) with a simple auditory detection task and a dichotic listening paradigm in which a voice was presented, lateralized, within a binaural white noise stream. Slower reaction times and a reduced right ear advantage might be expected at high (vs. low) altitude, due to hypoxia. The participants' performances on both tasks did not differ between conditions, revealing that these functions are resistant to short-term hypoxia in young healthy adults.