Effects of left bundle branch block on echocardiographic coronary flow assessment: A systematic review.

This systematic review investigates the diagnostic and prognostic utility of coronary flow reserve (CFR) assessment through echocardiography in patients with left bundle branch block (LBBB), a condition known to complicate the clinical evaluation of coronary artery disease (CAD). The literature search was performed on PubMed, EMBASE, Web of Science, Scopus, and Google Scholar, was guided by PRISMA standards up to March 2024, and yielded six observational studies that met inclusion criteria. These studies involved a diverse population of patients with LBBB, employing echocardiographic protocols to clarify the impact of LBBB on coronary flow dynamics. The findings emphasize the importance of CFR in stratifying cardiovascular risk and guiding clinical decision-making in patients with LBBB. Pooled results reveal that patients with LBBB and significant left anterior descending (LAD) artery stenosis exhibited a marked decrease in stress-peak diastolic velocity (MD = -19.03 [-23.58; -14.48] cm/s; p < .0001) and CFR (MD = -.60 [-.71; -.50]; p < .0001), compared to those without significant LAD lesions, suggesting the efficacy of stress echocardiography CFR assessment in the identification of clinically significant CAD among the LBBB population. This review highlights the clinical relevance of echocardiography CFR assessment as a noninvasive tool for evaluating CAD and stratifying risk in the presence of LBBB and underscores the need for standardized protocols in CFR measurement.

Balneotherapy and cortisol levels: an updated systematic review and meta-analysis.

The main objective of this review is to examine the impact of balneotherapy on serum and salivary cortisol concentrations. A systematic search was conducted in PubMed, Scopus, Web of Science, PEDro, and Google Scholar. The databases were screened from inception up until April 2024. After screening the scientific literature, 845 articles were retrieved and 17 studies, involving a total of 765 participants, were eventually included in the review. Among them, four were randomized controlled trials, five were non-randomized studies, and eight were pre-post studies with no control group. The evidence gathered in this review indicates a significant short-term reduction in cortisol levels in healthy individuals undergoing balneotherapy, particularly those experiencing high levels of stress. Conversely, in patients with rheumatic conditions (especially if elderly), increases in cortisol levels induced by balneotherapy can act as beneficial hormetic stress, reducing inflammatory mediators and improving pain and functional quality of life. The meta-analysis shows an overall trend of reduction in stress hormone levels, more pronounced in the intervention group undergoing mud-balneotherapy compared to the control group, a finding that, however, does not reach statistical significance (g=-0.11 [95% CI: -0.30; 0.08]; p > 0.05). Current scientific evidence demonstrates that balneotherapy has a positive impact on the regulation of cortisol levels. The regulation of the hypothalamic-pituitary-adrenal axis and the beneficial effects observed on health parameters and quality of life allow mud-balneotherapy to be classified as eustressful stimuli useful in preventing stress-related pathologies in healthy individuals and in alleviating symptoms in patients with chronic conditions. Future research on the topic is advised.

Defining a new perspective in Environmental Health: the healing environment.

With pollution-related health problems on the rise, the focus of modern Environmental Health (EH) has mostly been placed on toxicology and exposure science. Despite the importance of toxicological aspects, the environment should be studied not only to identify pollution-related hazards, but also to investigate potentially therapeutic and health-enhancing effects of its elements. Generally speaking, it is possible to benefit from a natural environment with a full-immersion experience or with a single-element interaction. Recently, scientific evidence is accumulating on the beneficial effects of natural settings for well-being promotion and psycho-physical health, especially for stress reduction and prevention of stress-related conditions. In light of these considerations, the paradigm of EH can change: the environment we live in should be considered not only as a precious resource to be protected against pollution (thus preventing the consequent health hazards), but, in a proactive vision, also as a potential source of elements capable of actively maintaining and promoting health and well-being.

Effects of forest bathing (shinrin-yoku) on individual well-being: an umbrella review.

The aim of this review was to outline the most relevant benefits of forest bathing for well-being promotion. This study was designed as an umbrella literature review. Medline (via PubMed), EMBASE, Web of Science, Cochrane Library, Google Scholar, Scopus, PsycINFO, CINAHL and the DOAJ were systematically searched for relevant reviews up to February 2021. After article selection, 16 systematic reviews met inclusion criteria. Overall, the best available evidence supports the use of forest bathing as a complementary practice for the promotion of psychophysical well-being, whereas evidence for its use as a therapeutic practice for the improvement of organic diseases needs to grow before clear and specific clinical indications can be formulated. The positive impact of forest bathing on individual quality of life, along with its favorable cost-effectiveness profile, may justify its possible adoption for public health strategies of well-being promotion. Further investigations on the topic are advised.

Clinical efficacy of medical hydrology: an umbrella review.

The aim of this research was to summarize available scientific evidence on the efficacy of medical hydrology for the management of any health condition. The search was conducted on 26th March 2021, in the following databases: Medline (via PubMed), EMBASE, Web of Science, Cochrane Library, and Google Scholar. All relevant literature reviews investigating the clinical efficacy of interventions characterized by the use of natural mineral waters and muds were included. The quality of studies was assessed with the "AMSTAR 2" tool. After article screening, 49 reviews were included in this work. Overall, retrieved scientific evidence suggests that spa therapy is beneficial for patients affected by some specific musculoskeletal conditions, with improvements potentially lasting up to 9 months. Moreover, balneotherapy can be an integrative support for the management of chronic venous insufficiency and some inflammatory skin diseases like psoriasis. The role of spa therapy in rehabilitation appears relevant as well. More limited, although interesting evidence exists for inhalation and hydropinic therapies. Globally, retrieved evidence suggests that, besides individual wellbeing, medical hydrology can be useful for public health. In particular, higher-quality studies seem to support the integrative use of spa-related interventions for conditions like osteoarthritis, fibromyalgia, low back pain of rheumatic origin, and chronic venous insufficiency. However, the body of evidence has some limitations and further clinical trials should be designed for each relevant application to consolidate and expand acquired knowledge.

Respiratory rehabilitation for post-COVID19 patients in spa centers: first steps from theory to practice.

With this correspondence, we would like to briefly outline a practical perspective about a possible integrative and effective management in spa settings of COVID-19 long-term sequelae, with a keen focus on post-infective lung damage and fibrosis, which is expected to become epidemiologically relevant in the general population. In order to outline a standard/baseline model of care, we think that it can be useful to refer to already existing rehabilitative plans with a long-standing tradition in Italy, such as those ones prescribed for work-related respiratory diseases like pneumoconiosis, in which long-term outcomes share some clinical characteristics with post-infective lung fibrosis. Such programs include diagnostic procedures (spirometry, ECG, blood tests) and treatments like respiratory physio-kinesiotherapy and postural drainage of the lungs; mechanical pulmonary ventilation for rehabilitative purposes, with or without drugs, along with standard medical and, when required, oxygen therapy; inhalation therapies with mineral waters; physical activity and psychological support. In conclusion, we believe that spa facilities can be a proper setting for respiratory rehabilitation and that already existing programs employed in occupational medicine can be a good starting point to plan rehabilitative strategies for post-COVID-19 patients. In particular, health spa centers can be useful not only to offer tailored programs of physical rehabilitation but also to provide patients with a psychologically supportive and health-promoting environment. Further studies on the topic are advised to properly assess and quantify with adequate outcome measurements the beneficial effect of a spa-based rehabilitative program in post-COVID-19 patients.

Effects of forest bathing (shinrin-yoku) on levels of cortisol as a stress biomarker: a systematic review and meta-analysis.

Forest bathing is a traditional practice characterized by visiting a forest and breathing its air. This review aims to investigate the effects of forest bathing on levels of salivary or serum cortisol as a stress biomarker in order to understand whether forest bathing can reduce stress. Medline/PubMed, Embase, Scopus, Web of Science, Cochrane Library, and Google Scholar were systematically searched for relevant articles. The quality of included trials was assessed following the criteria of the NIH dedicated tools. Afterwards, a qualitative and quantitative synthesis of retrieved evidence was performed. A total of 971 articles were screened; 22 of them were included in the systematic review and 8 in the meta-analysis. In all but two included studies, cortisol levels were significantly lower after intervention in forest groups if compared with control/comparison groups, or a significant pre-post reduction of cortisol levels was reported in the forest groups. The main results of the meta-analysis showed that salivary cortisol levels were significantly lower in the forest groups compared with the urban groups both before (MD = - 0.08 µg/dl [95% CI - 0.11 to - 0.05 µg/dl]; p < 0.01; I2 = 46%) and after intervention (MD = - 0.05 µg/dl [95% CI - 0.06 to - 0.04 µg/dl]; p < 0.01; I2 = 88%). Overall, forest bathing can significantly influence cortisol levels on a short term in such a way as to reduce stress, and anticipated placebo effects can play an important role in it. Further research is advised because of the limited available data.

Therapeutic efficacy of orally administered pollen for nonallergic diseases: An umbrella review.

Pollen has been used for centuries as a tonic and a multipurpose remedy in traditional medicine. The present umbrella review aims to qualitatively assess the therapeutic efficacy of orally administered pollen in the management of nonallergic diseases. MEDLINE via PubMed, Embase, Scopus, Web of Science, CINAHL, Cochrane Library, and Google Scholar were systematically searched for relevant systematic reviews and meta-analyses. Articles were independently screened and selected, then quality of evidence of included studies was evaluated with a dedicated NIH tool. Retrieved evidence was critically appraised and discussed. Two hundred four articles were found and, after selection process, five systematic reviews were included in the present work, including one with a meta-analysis. Evidence from these reviews supports the use of grass pollen extracts for symptomatic benign prostatic hyperplasia (BPH) and chronic prostatitis (CP). Additional preliminary evidence on the topic indicates the potential use of grass pollen extracts for vasomotor symptoms in women. Overall, results of the present review suggest that flower pollen extracts may be useful as a complementary remedy for the management of BPH, CP, and vasomotor symptoms. Evidence regarding bee pollen is too limited to draw any conclusion on its clinical efficacy. Further studies are needed.

Effects of balneotherapy and spa therapy on quality of life of patients with knee osteoarthritis: a systematic review and meta-analysis.

Knee osteoarthritis (OA) is a degenerative disease which is expected to become one of the leading causes of disability by the next years. This work aims to assess if balneotherapy and spa therapy can significantly improve Quality of Life (QoL) of patients with knee OA. Medline via PubMed, Scopus, Web of Science, Cochrane Library, and PEDro were systematically searched for articles about trials involving patients with knee OA and measuring the effects of balneotherapy and spa therapy on study participants' QoL with validated scales. A qualitative and quantitative syntheses were performed. Seventeen studies were considered eligible and included in the systematic review. Fourteen trials reported significant improvements in at least one QoL item after treatment. Ten studies were included in quantitative synthesis. When comparing balneological interventions with standard treatment, results favored the former in terms of long-term overall QoL [ES = - 1.03 (95% CI - 1.66 to - 0.40)]. When comparing balneological interventions with sham interventions, results favored the former in terms of long-term pain improvement [ES = - 0.38 (95% CI - 0.74 to - 0.02)], while no significant difference was found when considering social function [ES = - 0.16 (95% CI - 0.52 to 0.19)]. In conclusion, even though limitations must be considered, evidence shows that BT and spa therapy can significantly improve QoL of patients with knee OA. Moreover, reduction of drug consumption and improvement of algofunctional indexes may be other beneficial effects. Further investigation is needed because of limited available data.

Effects of balneotherapy and spa therapy on levels of cortisol as a stress biomarker: a systematic review.

Balneotherapy and spa therapy are well-known practices, even though limited evidence has been produced about their biological effects. This systematic review primarily aims at assessing if balneotherapy, mud/peloid therapy, and spa therapy may influence cortisol levels. Secondarily, it aims at understanding if these interventions may improve stress resilience. PubMed/Medline, Embase, and Cochrane Library were searched for relevant articles in English or Italian about studies involving healthy and sub-healthy subjects or patients with a diagnosed disease about effects of balneotherapy, mud/peloid therapy, and spa therapy on serum and salivary cortisol levels. Fifteen studies involving 684 subjects were included. Five studies investigated biological effects of balneotherapy alone. Two of them reported significant changes of cortisol levels in healthy participants. The other three studies reported no significant variations in patients with rheumatic conditions. No studies investigated biological effects of mud/peloid therapy alone. Ten studies investigated biological effects of spa therapy with or without included mud/peloid therapy, and in all but two studies, significant variations of cortisol levels were reported. Our main findings suggest that balneotherapy may have the potential to influence cortisol levels in healthy subjects, in such a way as to improve stress resilience. Spa therapy with or without included mud/peloid therapy demonstrated the same potential to influence cortisol levels also in sub-healthy subjects and in patients with a diagnosed disease. Therefore, balneotherapy and spa therapy may be considered as useful interventions for the management of stress conditions. Further investigation is needed because of limited available data.

Mud therapy and skin microbiome: a review.

Recent findings highlight the role of skin microbiome in modulating immune function and inflammatory response. This systematic review aims to investigate the effects of muds, already used for the treatment of several rheumatic and dermatologic conditions, on skin microbiome. Medline via PubMed, Embase, Cochrane Library, and Google Scholar were searched for articles about the effects of therapeutic muds on microorganisms of skin microbiome. Five studies were included in the review and critically appraised. Limited data suggest that muds have various properties that may explain their action on skin microbiome, with different effects on commensal and pathogenic microorganisms. Further investigation on this topic is needed to better characterize the effects of different muds on skin microbiome, thus possibly extending their indications.

[Prevalence of artificial sweeteners in multivitamin supplements registered in Italy.] / Prevalenza dei dolcificanti artificiali negli integratori multivitaminici registrati in Italia.

This study aims to evaluate the prevalence of artificial sweeteners in multivitamin supplements registered in Italy. The National Registry of Dietary Supplements was examined in December 2023 to extract data relating to supplements identified as "multivitamins". Then, for each supplement the specific composition was searched online. 179 multivitamin products were identified, and for 96 of them it was possible to access the complete list of their ingredients online. Among these, 47 supplements (49%) do not contain added sugars, while the remaining 49 (51%) have one or more sweeteners in their composition (29 of which have at least two among the ingredients). Among the 49 supplements in the second group, the greatest prevalence of artificial sweeteners is found in effervescent tablets (100%), soluble powders (100%) and gummies or chewing tablets (63%), while the lowest prevalence was detected in capsules/tablets to be swallowed (19%). In relative terms, aspartame and sucralose are mostly found in effervescent tablets and diluted powders, but other formulations are not always free of these substances. In conclusion, the significant presence of artificial sweeteners in multivitamin supplements requires attention, especially considering the long-term health impact. Consumer awareness and medical evaluation are crucial for an informed choice and the protection of health.

Evaluating qigong as integrative support for COVID-19 and Long-COVID-19 rehabilitation: a systematic review.

Introduction: Amidst the ongoing global impact of COVID-19 on public health, there is an increasing focus on holistic strategies encompassing integrative therapies and rehabilitation techniques, particularly in addressing the challenges posed by Long-COVID-19. This review investigates the potential of Qigong, an ancient Chinese practice characterized by gentle movements, controlled breathing, and meditative elements, within the context of COVID-19. Methods: A systematic search of PubMed, EMBASE, Web of Science, Scopus, and Google Scholar was conducted to identify pertinent clinical studies. Results: Following thorough database scrutiny, nine studies were identified as meeting the eligibility criteria. Across the spectrum of COVID-19 severity, individuals engaging in qigong practice exhibited notable enhancements in both physical and psychological wellbeing, evidenced by ameliorated respiratory symptoms, reduced anxiety levels, enhanced sleep quality, bolstered mental wellbeing, and augmented health-related quality of life. Moreover, qigong training, whether employed independently or in conjunction with other therapies, demonstrated beneficial effects on Long-COVID-19 symptoms, encompassing persistent respiratory issues, dizziness, sleep disturbances, and compromised health-related quality of life. Discussion: This review underscores the necessity for further investigation to quantify and standardize the contribution of Qigong to COVID-19 recovery and rehabilitation. Such endeavors aim to integrate this accessible and low-impact practice into public health strategies and comprehensive treatment regimens. Systematic review registration: The review protocol was registered in the Open Science Framework under the following doi: 10.17605/OSF.IO/7K5X6 (URL: https://osf.io/7k5x6).

Effects of Static Meditation Practice on Blood Lipid Levels: A Systematic Review and Meta-Analysis.

This review aims to delineate the potential impact of static meditation practice on cholesterol and triglyceride levels. PubMed, EMBASE, Web of Science, Cochrane Library, and Google Scholar were systematically screened up until December 2023 to identify pertinent studies. After searching the scientific literature, 16 clinical studies (11 trials and 5 observational experiments) met the criteria for inclusion, involving a total of 1147 participants. In general, Ayurvedic-based meditation techniques were predominantly associated with lower total cholesterol levels, mindfulness-based techniques demonstrated benefits in both total cholesterol and triglyceride levels, and Eastern meditation techniques with spiritual origins were primarily linked to improved serum concentrations of HDL cholesterol. Study participants mostly engaged in meditation on a daily basis, often practicing it once or even twice a day, spanning a duration ranging from a few weeks to several months. The meta-analysis shows an association between meditation practice in healthy or sub-healthy adults and reduced cholesterol levels, with an average decrease of approximately -14 mg/dL (MD = -13.91 [-23.35; -4.47] mg/dL; p = 0.02), alongside favorable and even more pronounced impacts on triglyceride levels (MD = -32.56 [-48.44; -16.68] mg/dL; p < 0.01). In summary, regular engagement in static meditation practices can be associated with lower triglyceride and, to a lesser extent, cholesterol levels. Further studies on the topic are recommended to better investigate the relationship between meditation practice and physiological parameters.

Validation Study of the Spanish Version of the Measure of Happiness (MH) Questionnaire.

This study aims to validate the Spanish version of the Measure of Happiness (MH), a questionnaire that identifies the specific areas of an individual's life that are related to experienced happiness. The sample consisted of 458 Spanish native speakers (65.7% women, 34.3% men; mean age = 24.14, SD = 8.45) and was divided into two groups: Sample 1 (N = 226) underwent Exploratory Factor Analysis (EFA), and Sample 2 (N = 232) underwent Confirmatory Factor Analysis (CFA). The convergent and discriminant validity of the Spanish version of the MH and its subscales was assessed by calculating Pearson correlations between the MH factors and the Subjective Happiness Scale, the State-Trait Anxiety Inventory, the Beck Depression Inventory, and the WHOQOL-BREF. The EFA and CFA confirmed the original five-factor structure of the MH questionnaire. The only difference in the Spanish version of the MH is that Item 4, which was originally assigned to Factor 2 "Financial Status", was reallocated to Factor 1 "Psychophysical Status". The reliability of the Spanish version of the MH questionnaire was also confirmed, and the factors correlated in the expected direction with the constructs of quality of life, happiness, anxiety, and depression. The MH questionnaire demonstrated excellent psychometric properties among Spanish respondents.

Improved Cardiovascular Effects of a Novel Pomegranate Byproduct Extract Obtained through Hydrodynamic Cavitation.

The healthy properties of pomegranate fruit, a highly consumed food, have been known for a long time. However, the pomegranate supply chain is still rather inefficient, with the non-edible fraction, whose weight is roughly half the total and is endowed with plenty of valuable bioactive compounds, either disposed of or underutilized. A novel extract obtained from non-edible byproducts (called PPE), using hydrodynamic cavitation, a green, efficient, and scalable technique, was investigated for its cardiovascular effects in vivo. PPE showed efficacy in an acute phenylephrine (PE)-induced hypertensive rat model, similar to the extract of whole fruit (PFE) obtained using the same extractive technique, along with good intestinal bioaccessibility after oral administration. Finally, when chronically administered for 6 weeks to spontaneously hypertensive rats, PPE was shown to significantly contain the increase in systolic blood pressure, comparable to the reference drug Captopril, and at a dose remarkably lower than the reported effective dose of ellagic acid. The extract from the non-edible fraction of the pomegranate fruit also showed good anti-inflammation and anti-fibrotic effects. The findings of this study, along with the extraction technique, could contribute to enhancing the value of the pomegranate supply chain, relieve the related environmental burden, and potentially improve public health.

Olfactory training with essential oils for patients with post-COVID-19 smell dysfunction: A case series.

Introduction: It is estimated that up to one third of COVID-19 patients can develop long-lasting smell dysfunction. Viral infections, especially COVID-19, can cause anosmia through different pathomechanisms, and different strategies have been proposed for effectively managing post-COVID-19 olfactory dysfunction in clinical practice, with olfactory training being recommended as a first-line treatment option. Methods: This report describes a non-consecutive series of clinical cases. After COVID-19, eight cases (5 females, 3 males) of adult patients with long-lasting (3+ months) post-viral smell dysfunction followed a 30-day olfactory training protocol with a set of plant-derived essential oils. At baseline and at the end of the treatment, the patients were administered the Assessment of Self-reported Olfactory Functioning (ASOF) questionnaire, an inventory used to measure olfactory dysfunction and health-related quality of life. Results: For any of the outcomes assessed with the ASOF scale, a significant improvement from baseline was reported, even though mean value ameliorations were more pronounced for olfactory function per se (Subjective Olfactory Capability: from 3.6 to 5.6 out of 10; Self-Reported capability of Perceiving specific odors: from 1.8 to 3.0 out of 5), rather than for health-related quality of life (Olfactory-Related Quality of life: from 2.9 to 3.9 out of 6). Conclusions: It was observed that patients with long-lasting COVID-19-related smell dysfunction improved after a 30-day olfactory training protocol. Further controlled clinical studies would be useful to better investigate the role of olfactory training in patients with postviral smell dysfunction.

Silence and its effects on the autonomic nervous system: A systematic review.

This systematic review explores the influence of silence on the autonomic nervous system. The Polyvagal Theory has been used as a reference model to describe the autonomic nervous system by explaining its role in emotional regulation, social engagement, and adaptive physiological responses. PubMed, Scopus, PsycInfo, EMBASE, and Google Scholar were systematically searched up until July 2023 for relevant studies. The literature search yielded 511 results, and 37 studies were eventually included in this review. Silence affects the autonomic nervous system differently based on whether it is inner or outer silence. Inner silence enhances activity of the ventral vagus, favoring social engagement, and reducing sympathetic nervous system activity and physiological stress. Outer silence, conversely, can induce a heightened state of alertness, potentially triggering vagal brake removal and sympathetic nervous system activation, though with training, it can foster inner silence, preventing such activation. The autonomic nervous system response to silence can also be influenced by other factors such as context, familiarity with silence, presence and quality of outer noise, and empathy.

Partial articular fractures of the radial head: How to understand them using computed tomography.

BACKGROUND: The optimal treatment for different types of displaced partial articular radial head fractures is still debatable. Fractures involving the articulating portion of the radial head can be defined also as bi-articular and are often underestimated. Moreover, the complete loss of periosteal contact between fracture fragments is a marker of instability. Available classifications do not emphasize these aspects. The purpose of the present study is to describe two assessment methods to identify fractures involving the articulating portion of the radial head and complete loss of periosteal contact with the help of 2D-3D CT scan. The second purpose is to propose a classification of the displaced partial articular radial head fractures basing on these two assessment methods. HYPOTHESIS: We hypothesize that the proposed classification is reliable. PATIENTS AND METHODS: By observing the position of the bicipital tuberosity with respect to the ulna in reference to the coronal plane in the 3D-CT scan and by observing the location of the fracture fragment in the 2D axial scan, it is possible to understand if a displaced partial articular fracture involves the articulating portion of the radial head. Also, it is possible to understand the presence or absence of a complete loss of periosteal contact between the two fracture fragments by observing the coronal and sagittal 2D-CT scans. We identified 20 displaced partial articular radial head fractures on a series of 149 patients with radial head fractures. Following the above-described assessment methods, these 20 fractures were classified in three subgroups. Also, seven evaluators were asked to use these assessment methods to classify these 20 fractures in the three subgroups, in order to evaluate inter- and intra-observer agreement. RESULTS: Eight fractures involved the non-articulating portion of the radial head with complete loss of periosteal contact, eight involved the non-articulating portion of the radial head without complete loss of periosteal contact, and four involved the articulating portion of the radial head with or without complete loss of periosteal contact. The kappa for intra-observer reliability ranged from 0.46 to 0.84. The average kappa for inter-observer reliability was 0.570 (range: 0.526 to 0.676). The Kendall's coefficient for inter-observer concordance was 0.673. DISCUSSION: Prognostically, displaced partial fractures of the articulating portion of the radial head could differ from the other types, regardless of whether or not there is a complete loss of periosteal contact. Underestimating this fracture pattern can lead to poor results due to risk of forearm rotation blockage. Moreover, underestimating complete loss of periosteal contact in displaced partial fractures of the non-articulating portion of the radial head could lead to poor results. The described evaluation methods have moderate reliability, but can represent, along with other described methods, a good starting point to better understand and treat these insidious fractures. LEVEL OF EVIDENCE: III; retrospective study.