Carotid Intima-Media Thickness, Overweight, and Poor Sleep Quality in Commercial Airline Pilots: A Cross-Sectional Study.

Background Cardiovascular disease is a leading cause of premature career termination in commercial airline pilots (APs). In this cross-sectional study, we sought to investigate the relationship between intima-media thickness (IMT), a marker of subclinical atherosclerosis, and cardiovascular risk factors in APs, focusing on overweight status and sleep quality. Methods A total of 140 male APs were categorized into four groups based on body mass index (BMI) and Pittsburgh Sleep Quality Index (PSQI) score: overweight poor sleepers (OW-PS), overweight good sleepers (OW-GS), normal weight poor sleepers (NW-PS), and normal weight good sleepers (NW-GS). IMT was quantified in the common carotid artery (CCA) and carotid bulb using ultrasound, yielding a composite IMT (IMTcom) measure. Common cardiovascular risk factors were assessed in all participants. Results The prevalence of overweight and poor sleep quality was 43.6% and 32.9%, respectively. The OW-PS group had significantly higher age, heart rate, total cholesterol, and low-density lipoprotein (LDL) cholesterol compared to other groups (p<0.05). Overweight pilots, regardless of sleep quality, had increased IMTcom compared to normal-weight pilots (p<0.001). Age and LDL cholesterol were independent predictors of IMTcom in the OW-PS and OW-GS groups (p<0.05). Conclusions Overweight status, irrespective of sleep quality, is associated with increased IMT in APs, suggesting a higher burden of subclinical atherosclerosis. Interventions focused on reducing LDL cholesterol levels and managing age-related cardiovascular risk factors could be advantageous in mitigating the risk of atherosclerotic vascular disease in overweight pilots.

Untargeted blood serum proteomics identifies novel proteins related to neurological recovery after human spinal cord injury.

BACKGROUND: The discovery of new prognostic biomarkers following spinal cord injury (SCI) is a rapidly growing field that could help uncover the underlying pathological mechanisms of SCI and aid in the development of new therapies. To date, this search has largely focused on the initial days after the lesion. However, during the subacute stage of SCI (weeks to months after the injury), there remains potential for sensorimotor recovery, and numerous secondary events develop in various organs. Additionally, the confounding effects of early interventions after the injury are less likely to interfere with the results. METHODS: In this study, we conducted an untargeted proteomics analysis to identify biomarkers of recovery in blood serum samples during the subacute phase of SCI patients, comparing those with strong recovery to those with no recovery between 30 and 120 days. We analyzed the fraction of serum that is depleted of the most abundant proteins to unmask proteins that would otherwise go undetected. Linear models were used to identify peptides and proteins related to neurological recovery and we validated changes in some of these proteins using Enzyme-linked Immunosorbent Assay (ELISA). RESULTS: Our findings reveal that differences in subacute recovery after SCI (from 30 to 120 days) are associated with an enrichment in proteins involved in inflammation, coagulation, and lipid metabolism. Technical validation using commercial ELISAs further confirms that high levels of SERPINE1 and ARHGAP35 are associated with strong neurological recovery, while high levels of CD300a and DEFA1 are associated with a lack of recovery. CONCLUSIONS: Our study identifies new candidates for biomarkers of neurological recovery and for novel therapeutic targets after SCI.

Microbiota Analysis of Ejaculated Honey Bee Drone Semen and the Effect of Semen Collection Method on Bacterial Loads.

Artificial insemination in queen honey bees is the only tool that provides complete control over mating for research and breeding purposes, making it essential in genetic improvement and conservation programs in this species. The aims of this study were to characterize drone semen bacterial loads by culture-dependent and independent methods and to describe their variation depending on the method of semen collection, the colony and the apiary. In the first experiment, the bacterial loads of semen collected from the seminal vesicles or from ejaculates was studied using culture-dependent methods. The collection method had a significant influence on the overall bacterial count in semen. Out of the 42 semen samples analyzed, 26 (61.9%) tested positive for bacterial isolation. This encompassed the entirety of samples obtained from the seminal vesicles (21 of 21), whereas only 23.8% of those derived from ejaculates (5 out of 21) showed bacterial isolation. In the second experiment, next-generation sequencing techniques were used to describe the microbiome of ejaculated drone semen for the first time. The most abundant phyla were Proteobacteria, Firmicutes, Bacteroidota and Actinobacteriota, while the most abundant genera were Lactobacillus, Staphylococcus, Prevotella, Alloprevotella and Streptococcus. The results showed that the apiary had a significant effect on the community structure composition and abundance of the seminal microbiota, and significative differences in abundance were observed for the genera Sphingomonas, Methylobacterium-Methylorubrum, Bifidobacterium and Alloprevotella. Significant differences were also observed in the richness of the microbiota between apiaries and colonies.

Brushing Away the Blues: Self-Reported Oral Hygiene Practices Are Associated With Mild Depressive Symptoms in Airline Pilots.

Introduction Airline pilots are susceptible to mental health issues, with depression prevalence ranging from 1.9% to 12.6%. Recent research in the general population indicates a potential link between depression and oral health. In this cross-sectional study, we sought to investigate the association between self-reported oral hygiene practices and depressive symptoms among airline pilots. Methods One hundred actively working male airline pilots of Caucasian descent voluntarily enrolled in the study during routine occupational health visits. Depressive symptoms were assessed using the Beck Depression Inventory II (BDI-II). Self-reported oral hygiene practices, including toothbrushing frequency and mouthwash usage, were examined. Univariable and multivariable logistic regression analyses were performed to investigate associations between depressive symptoms and oral hygiene practices. Results Twelve pilots (12%) demonstrated mild depressive symptomatology (BDI-II scores 14-19). Pilots with mild depression reported significantly lower rates of brushing teeth twice or more per day (33.3% vs. 80.7%) and higher rates of rarely brushing (16.7% vs. 1.1%) compared to those with minimal depressive symptoms (p < 0.001). Nonuse of mouthwash was more prevalent among pilots with mild depression (66.6% vs. 23.9%, p = 0.008). Multivariable logistic regression analysis revealed that pilots who rarely brushed their teeth (adjusted odds ratio (OR) = 14.6; 95% confidence interval (CI) = 1.3-197.9; p < 0.05) or did not use mouthwash (adjusted OR = 5.7; 95% CI = 1.4-25.2; p < 0.05) had significantly higher odds of mild depressive symptoms. Conclusions Self-reported oral hygiene habits may serve as a proxy indicator for mild depressive symptoms among airline pilots. Incorporating oral health assessments into routine aeromedical examinations could provide a practical method of identifying pilots at risk for depression, supporting timely interventions and enhancing flight safety.

Prospective Study of Homologous Recombination Repair Gene Mutation Prevalence in Patients With Advanced Prostate Cancer From Latin America: Challenges and Future Approaches.

PURPOSE: The prevalence of homologous recombination repair gene mutations (HRRm) in patients with metastatic castration-resistant prostate cancer (mCRPC) in Latin America and the Caribbean (LAC) is unknown. Prevalence of homologous Recombination repair (HRR) gene mutatiOns in patientS with metastatic castration resistant ProstatE Cancer in LaTin America (PROSPECT) aimed to determine this prevalence and to describe the demographic and clinical characteristics of the participants. MATERIALS AND METHODS: This was a prospective, cross-sectional, multicenter study across 11 cancer centers in seven LAC countries. After informed consent, all eligible participants underwent genomic testing by provided blood samples for germline HRR testing; they also provided PC tissue blocks if available for somatic HRR testing. RESULTS: Between April 2021 and April 2022, 387 patients (median age, 70 years [49-89], 94.3% Eastern Cooperative Oncology Group 0-1) with mCRPC were enrolled in the study. Almost 40% of them had a family history of cancer, and the overall time from their initial PC and mCRPC diagnosis was 3 years and 1 year, respectively. The overall prevalence of germline HRRm was 4.2%. The mutations detected included the genes CHEK2 (n = 4, 1%), ATM (n = 3, 0.8%), BRCA2 (n = 3, 0.8%), BRIP1 (n = 2, 0.5%), RAD51B (n = 2, 0.5%), BRCA1 (n = 1, 0.3%), and MRE11 (n = 1, 0.3%). The prevalence of somatic HRRm could not be assessed because of high HRR testing failure rates (79%, 199/251) associated with insufficient DNA, absence of tumor cells, and poor-quality DNA. CONCLUSION: Despite the study's limitations, to our knowledge, PROSPECT was the first attempt to describe the prevalence of HRRm in patients with PC from LAC. Notably, the germline HRRm prevalence in this study was inferior to that observed in North American and European populations. The somatic HRR testing barriers identified are being addressed by several projects to improve access to HRR testing and biomarker-based therapies in LAC.

Efficacy and Safety of Two Chondroprotective Supplements in Patients With Knee Osteoarthritis: A Randomized, Single-Blind, Pilot Study.

Background: Hyaluronic acid (HA), glucosamine (Glc), and chondroitin sulfate (CS) are key ingredients commonly incorporated into dietary chondroprotective supplements for the management of osteoarthritis (OA). Despite their widespread use, there is a paucity of published data regarding their efficacy and safety, necessitating rigorous investigation in clinical settings. To address this knowledge gap, we conducted a randomized, single-blind pilot study to evaluate the effects of two commercially available multi-ingredient supplements on patients with mild-to-moderate knee OA. Methods: A total of 51 patients diagnosed with mild-to-moderate knee OA were enrolled in a four-week randomized study and allocated equally (1:1:1 ratio) into three groups: a control group (n = 17) that received no treatment, an HA group (n = 17) given Syalox® 300 Plus (1 tablet/day) containing HA (300 mg) and Boswellia serrata extract (100 mg), and a Glc + CS group (n = 17) given Cartijoint® Forte (1 tablet/day) containing Glc (415 mg), CS (400 mg), and curcuminoids from rhizomes of Curcuma longa L (50 mg).Physicians conducting evaluations were blinded to group assignments, whereas patients were not. All participants underwent assessments of pain relief, functional capacity improvement, and serum adropin levels, an emerging biomarker of knee OA, at baseline and after the four-week intervention period. Results: Both the HA and the Glc + CS groups exhibited improvements at the end of the study relative to baseline, with statistically significant differences (p < 0.05) observed in pain at rest, pain during movement, range of motion, and the overall Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores, including its pain, stiffness, and physical function subscales. Notably, the HA group outperformed the Glc + CS group in the alleviation of pain at rest, pain during movement, and on the WOMAC pain subscale, with all differences being statistically significant (p < 0.05). Additionally, both groups showed a significant elevation in serum adropin levels from baseline (p < 0.05), with the HA group experiencing a more substantial increase when compared to the Glc + CS group (p < 0.05). Both supplements showed a limited number of treatment-emergent adverse events. Conclusion: Oral supplementation with either HA or Glc + CS demonstrated potential benefits for managing symptoms of mild-to-moderate knee OA. Notably, HA supplementation was associated with greater improvements in pain relief and higher elevations in serum adropin levels compared to Glc + CS supplementation. However, larger-scale and longer-term studies are necessary to further evaluate the safety and efficacy of these dietary supplements within the clinical management arsenal for knee OA.

Phenotypic and genotypic antimicrobial resistance of Listeria spp. in Spain.

Listeriosis is a zoonotic disease caused by Listeria monocytogenes and Listeria ivanovii. The genus Listeria currently includes 27 recognized species and is found throughout the environment. The number of systematic studies on antimicrobial resistance in L. monocytogenes isolates from domestic farms using antimicrobial substances is limited. Importantly, dairy ruminant farms are reservoir of hypervirulent lineage I L. monocytogenes isolates, previously associated with human clinical cases. Considering that the classes of antibiotics used in food-producing domestic animals are frequently the same or closely related to those used in human medicine, studies about the impact of antibiotic use on the acquisition of antibiotic resistance in Listeria spp. in domestic animal farms are, therefore, of high importance. Here, susceptibility to 25 antibiotics was determined. Eighty-one animal-related, 35 food and 21 human pathogenic Listeria spp. isolates and 114 animal-related non-pathogenic Listeria spp. isolates were tested. Whole genome sequencing data was used for molecular characterization. Regarding L. monocytogenes, 2 strains from the clinical-associated linage I showed resistance to erythromycin, both related to dairy ruminants. Acquired resistance to one antibiotic was exhibited in 1.5% of L. monocytogenes isolates compared with 14% of non-pathogenic Listeria spp. isolates. Resistance to tetracycline (7.9%), doxycycline (7.9%), penicillin (4.4%), and ampicillin (4.4%) were the most frequently observed in non-pathogenic Listeria spp. While resistance to two or more antibiotics (5.6%) was most common in Listeria spp., isolates, resistance to one antibiotic was also observed (1.6%). The present results show that non-pathogenic Listeria spp. harbour antimicrobial resistance genes.

Occupational Health Challenges for Aviation Workers Amid the Changing Climate: A Narrative Review.

Although there are many forecasts regarding the impact of climate change on the aviation sector, a critical but frequently neglected dimension is the occupational safety risks faced by aviation professionals. This narrative review explores the potential impacts of the changing climate on the health and safety of aviation personnel. Furthermore, we examine the significance of resilience in helping these workers adapt and effectively manage climate-related challenges in their professional lives. Climate change poses increasing threats to the well-being of flight personnel through elevated temperatures, heightened ultraviolet radiation exposure, increased mental workload from extreme weather events, and other psychological stressors. Building resilience through workforce training, planning, and adaptation can reduce vulnerability. In future research, the iterative process of selecting measurement components to gauge the impact of climate change should balance feasibility, relevance for stakeholders, and accurately capturing exposure effects. For instance, while salivary cortisol measures stress biologically, assessments of depression or burnout may provide more nuanced insights on pilot health for industry decision-makers managing climate impacts. In conclusion, a strategic emphasis on enhancing the physical and psychological well-being of the aviation workforce is imperative for facilitating a more efficient adaptation within the sector. This is of paramount importance, considering the critical function that aviation serves in fostering human connectivity. Consequently, it is essential for regulatory bodies and policymakers to prioritize the safeguarding of employee health in the face of climate change challenges.

Serum Prestin and Otolin-1 Levels in Pilots of Helicopter-Based Emergency Medical Services: Potential Markers for Ear Injury.

INTRODUCTION: Occupational noise exposure is a major public health concern, impacting a large workforce worldwide. In this study, we sought to evaluate the serum concentrations of prestin, a cochlear protein that diminishes following noise exposure, and otolin-1, a protein secreted into the bloodstream subsequent to inner ear damage, among three diverse professional categories, each exposed to varying degrees of noise. Helicopter emergency medical service (HEMS) pilots and construction workers were considered high-risk groups due to their elevated exposure to occupational noise, whereas office workers were regarded as a low-risk group, reflecting their comparatively minimal noise exposure. METHODS: The study sample included 60 males, encompassing helicopter pilots, construction laborers, and office workers (n=20, each). Recruitment occurred during standard occupational health visits, with all participants presenting normal clinical audiograms. Serum levels of prestin and otolin-1 were measured in duplicate using commercially available immunoassays and compared across the three professional categories. RESULTS: HEMS pilots had the lowest mean serum prestin level at 211±27 pg/mL, followed by construction workers at 234±29 pg/mL, and office workers at 269±42 pg/mL (p<0.001, one-way analysis of variance), with all inter-group differences statistically significant (p<0.05, Tukey's post hoc tests). For otolin-1, HEMS pilots showed the highest mean at 216±20 pg/mL, with construction workers at 196±22 pg/mL, and office workers at 181±20 pg/mL (p<0.001, one-way analysis of variance). Statistically significant differences were found between HEMS pilots and both other groups for otolin-1 levels (p<0.05, Tukey's post hoc tests), but not between construction workers and office workers. CONCLUSIONS:  Serum concentrations of prestin and otolin-1 may differ among healthy individuals according to their occupational noise exposure and have the potential to act as indicators of subclinical inner ear injury. To substantiate these preliminary observations, incorporating exposure assessment, especially via direct measurements of noise and vibration exposure, would markedly improve the reliability of our findings.

Successful Management of Chronic Wounds by an Autophagy-Activating Magnetized Water-Based Gel in Elderly Patients: A Case Series.

Chronic wounds pose a significant threat to human health, particularly for the elderly, and require extensive healthcare resources globally. Autophagy, a key molecular player in wound healing, not only offers a defense against infections but also contributes to the deposition of the extracellular matrix during the proliferative phase. Additionally, it promotes the proliferation and differentiation of endothelial cells, fibroblasts, and keratinocytes. We have recently shown that applying magnetized saline water topically can trigger autophagy in intact skin. In this case series, we document the successful management of five non-infected, difficult-to-heal wounds in elderly patients using a topical autophagy-stimulating gel containing 95% magnetized saline water. The treated wounds included pressure ulcers, venous ulcers, and trauma-related injuries that had shown minimal or no improvement with standard wound therapies over a prolonged period. Application of the autophagy-stimulating gel promoted wound healing, as indicated by reduced fibrous and necrotic tissue, granulation tissue formation, re-epithelialization, and partial or complete wound closure. These preliminary case studies suggest that a topical gel containing magnetized saline water, which promotes autophagy, may aid healing of chronic wounds in elderly patients. Further investigation is warranted to explore the potential of this novel approach, as it may offer a valuable addition to the existing arsenal of wound care treatments for the aging population, particularly in addressing difficult-to-heal wounds.

Fracturas de la extremidad proximal de húmero en tres y cuatro fragmentos en el anciano: ¿Tratamiento conservador o artroplastia reversa? / Fractures of the humeral proximal extremity in three and four fragments in the elderly patient: conservative treatment or reverse arthroplasty?

OBJETIVO Comparar los resultados funcionales obtenidos en dos cohortes de pacientes ancianos tratados de forma quirúrgica (artroplastia reversa de hombro) versus tratamiento conservador. MATERIAL Y METODOS Estudio ambispectivo, no aleatorizado. Se incluyeron pacientes de 75 años o más, 15 tratados de forma quirúrgica y 16 siguieron tratamiento conservador. Seguimiento mínimo de 12 meses. A los 3, 6 y 12 meses de la fractura evaluamos las escalas ASES, Constant-Murley and Disabilities of the Arm, Shoulder and Hand score (DASH) y EVA. Además evaluamos los resultados radiológicos y las complicaciones. RESULTADOS No encontramos diferencias significativas para las escalas ASES, DASH, ni EVA .El grupo tratado de forma quirúrgica obtuvo a los 12 meses una puntuación media en la escala Constant mayor, diferencia estadísticamente significativa(75.1 þ/-10.3 vs. 51.9 þ/-12.4 p » 0.001). Además presentaban mayor rango de movilidad para flexión y rotación externa (128.9 þ/-17 versus 99.3 þ/-20.1 p » 0.001, and 35.7 þ/-13.9 vs. 23.4 þ/-15.5 p » 0.032). El 43% de los pacientes tratados mediante artroplastia reversa presentaban tuberosidades normoposicionadas y mejores resultados en las escalas versus pacientes con tuberosidades ausentes o malposicionadas. El grupo sometido a cirugía no presentó mayor tasa de complicaciones.CONCLUSIONES El tratamiento conservador en las fracturas de EPH en tres y cuatro fragmentos en pacientes ancianos ofrece buenos resultados en cuanto a dolor y funcionalidad en la mayoría de pacientes. Falta por definir qué pacientes por tener alta demanda funcional serían candidatos a tratamiento quirúrgico de entrada

OBJECTIVE This study compared functional results in two cohorts of older adults with three- and four-part proximal humeral fractures (PHFs) per Neer's classification treated with reverse shoulder arthroplasty (RSA) or nonoperative management. MATERIALS AND METHODS Ambispective, non-randomized study with two cohorts of patients aged 75 or older treated with RSA (n » 15) or nonoperative management (n » 16) with a minimum follow-up period of 12 months. We analyzed the American Shoulder and Elbow Surgeons (ASES), Constant-Murley, and Disabilities of the Arm, Shoulder, and Hand (DASH) scores and the visual analog scale (VAS) for pain at 3, 6, and 12 months. In addition, we recorded radiological findings and surgical complications. RESULTS The mean Constant-Murley score at 12 months was significantly higher for the RSA group (75.1 þ/- 10.3 vs. 51.9 þ/- 12.4, p » 0.001). There were no differences in ASES, DASH, and VAS scores. Statistically significant differences for flexion and external rotation in abduction favored the RSA group (128.9 þ/- 17.0 versus 99.3 þ/- 20.1, p » 0.001, and 35.7 þ/- 13.9 vs. 23.4 þ/- 15.5, p » 0.032, respectively). For the RSA group, tuberosity positioning was correct in 43% of subjects. These patients presented better scores than those with malpositioned or absent tuberosities but with no statistical significance. Complications in the surgical group were not higher. CONCLUSIONS Nonoperative treatment is a valid option regarding pain and functionality in elderly patients with three- and four-part PHFs. Characteristics of patients with high demands who may be candidates for the initial surgical treatment remain to be defined

Controlling Coxiella burnetii in naturally infected sheep, goats and cows, and public health implications: a scoping review.

Q fever is a worldwide zoonotic disease which domestic ruminants are the main source of infection for humans. This scoping review summarizes the control measures currently available to reduce Coxiella burnetii (Cb) infection in naturally infected sheep, goat and cattle herds. A total of 28 articles were included in the review. A lack of methodological standardization was noted in the articles analyzed. The results indicated that long-term vaccination in cows reduces bacterial excretion in milk and environmental contamination. In small ruminants, the results of vaccination in terms of efficacy are variable. In goats, there is a reduction in bacterial excretion, unlike in sheep, where a long-term vaccination program is necessary to reduce bacterial excretion. Moreover, the high persistence of viable Cb in the environment means that control measures for sheep are needed for several years. The use of antibiotics as a control measure in cows and sheep was not found to reduce excretion. However, the combination of vaccination with antibiotic therapy appears to have positive effects in small ruminants in terms of controlling outbreaks of Q fever. Hygiene and biosecurity measures are the basic means for controlling Cb infection on ruminant farms and ensuring public health.

Relation of Sleep Quality to a Panel of Plasma Cardiometabolic Markers in Airline Pilots: A Cross-Sectional Study.

Background Insomnia and poor sleep are leading modifiable risk factors for cardiovascular disease. Given the high susceptibility of airline pilots (APs) to sleep disturbances, we sought to investigate the hypothesis that poor sleep in this professional group correlates with alterations in plasma biochemical markers that would reflect critical aspects in the pathophysiology of cardiometabolic disorders. Methods In this preliminary cross-sectional study, we examined the relation of poor sleep to fourteen plasma biomarkers reflecting multiple cardiometabolic pathways in a convenience sample of 117 male APs. The Pittsburgh Sleep Quality Index (PSQI) was used to categorize the participants into good sleepers (n = 70, 59.8%; PSQI scores from 0 to 4) and poor sleepers (n = 47, 40.2%; PSQI scores of 5 or higher). The concentrations of biomarkers were compared between the two groups using both univariable and multivariable analyses. Results Compared to good sleepers, APs identified as poor sleepers exhibited significantly different levels of four plasma cardiometabolic biochemical markers in univariable analysis. However, in multivariable-adjusted analysis, only three biomarkers, adiponectin, fibroblast growth factor (FGF)-21, and growth differentiation factor (GDF)-15, remained independently associated with poor sleep. Conclusion Poor sleep quality in APs correlates with lower plasma concentrations of adiponectin and elevated levels of FGF-21 and GDF-15. Further longitudinal studies are required to elucidate the role of these biomarkers in the link between sleep disturbances and cardiometabolic risk in this professional group.

In Vitro Interaction between Mycoplasma agalactiae and Small Ruminants' Endogenous Bacterial Strains of Enterococcus spp. and Coagulase-Negative Staphylococcus.

Recently, an antimicrobial effect on Mycoplasma agalactiae (Ma), the main etiological agent of contagious agalactia (CA), was reported in vitro with strains of Enterococcus spp. from ovine and caprine milk. The aim of this work was to evaluate the interaction of Ma with the same Enterococcus spp. isolated from other anatomical locations (vagina) and other bacterial populations present in milk, such as coagulase-negative staphylococci (CNS). The vaginal Enterococcus strains and the raw milk CNS were isolated from sheep and goats. Experimental in vitro conditions were prepared to assess the growth of Ma with and without the presence of these strains. The selected vaginal strains were identified as Enterococcus (E.) hirae and E. mundtii, and the strains of CNS were identified as Staphylococcus petrasii. Different interactions of Ma with ovine and caprine wild vaginal strains of Enterococcus and dairy strains of CNS are described for the first time: Ma can grow exponentially during 15 h with the selected strains, although with certain strains, its optimal growth can be negatively affected (p < 0.05). The colonization and/or excretion of Ma could, therefore, be influenced by certain endogenous bacterial strains. Our results increase the knowledge about possible bacterial ecology dynamics surrounding CA.

Trophectoderm cells of human mosaic embryos display increased apoptotic levels and impaired differentiation capacity: a molecular clue regarding their reproductive fate?

STUDY QUESTION: Are there cell lineage-related differences in the apoptotic rates and differentiation capacity of human blastocysts diagnosed as euploid, mosaic, and aneuploid after preimplantation genetic testing for aneuploidy (PGT-A) based on concurrent copy number and genotyping analysis? SUMMARY ANSWER: Trophectoderm (TE) cells of mosaic and aneuploid blastocysts exhibit significantly higher levels of apoptosis and significantly reduced differentiation capacity compared to those of euploid blastocysts. WHAT IS KNOWN ALREADY: Embryos diagnosed as mosaic after PGT-A can develop into healthy infants, yet understanding the reasons behind their reproductive potential requires further research. One hypothesis suggests that mosaicism can be normalized through selective apoptosis and reduced proliferation of aneuploid cells, but direct evidence of these mechanisms in human embryos is lacking. Additionally, data interpretation from studies involving mosaic embryos has been hampered by retrospective analysis methods and the high incidence of false-positive mosaic diagnoses stemming from the use of poorly specific PGT-A platforms. STUDY DESIGN, SIZE, DURATION: Prospective cohort study performing colocalization of cell-lineage and apoptotic markers by immunofluorescence (IF). We included a total of 64 human blastocysts donated to research on Day 5 or 6 post-fertilization (dpf) by 43 couples who underwent in vitro fertilization treatment with PGT-A at IVI-RMA Valencia between September 2019 and October 2022. A total of 27 mosaic blastocysts were analyzed. PARTICIPANTS/MATERIALS, SETTING, METHODS: The study consisted of two phases: Phase I (caspase-3, n = 53 blastocysts): n = 13 euploid, n = 22 mosaic, n = 18 aneuploid. Phase II (terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL), n = 11 blastocysts): n = 2 euploid, n = 5 mosaic, n = 4 aneuploid. Following donation for research, vitrified blastocysts were warmed, cultured until re-expansion, fixed, processed for IF, and imaged using confocal microscopy. For each blastocyst, the following cell counts were conducted: total cells (DAPI+), TE cells (GATA3+), inner cell mass (ICM) cells (GATA3-/NANOG+), and apoptotic cells (caspase-3+ or TUNEL+). The incidence of apoptosis was calculated for each blastocyst by dividing the number of caspase-3+ cells (Phase I) or TUNEL+ cells (Phase II) by the number of TE or ICM cells. Statistical analysis was performed according to data type and distribution (P < 0.05 was considered statistically significant). MAIN RESULTS AND THE ROLE OF CHANCE: Phase I: Mosaic blastocysts displayed a similar number of total cells (49.6 ± 15 cells at 5 dpf; 58.8 ± 16.9 cells at 6 dpf), TE cells (38.8 ± 13.7 cells at 5 dpf; 49.2 ± 16.2 cells at 6 dpf), and ICM cells (10.9 ± 4.2 cells at 5 dpf; 9.7 ± 7.1 cells at 6 dpf) compared to euploid and aneuploid blastocysts (P > 0.05). The proportion of TE cells retaining NANOG expression increased gradually from euploid blastocysts (9.7% = 63/651 cells at 5 dpf; 0% = 0/157 cells at 6 dpf) to mosaic blastocysts (13.1% = 104/794 cells at 5 dpf; 3.4% = 12/353 cells at 6 dpf) and aneuploid blastocysts (27.9% = 149/534 cells at 5 dpf; 4.6% = 19/417 cells at 6 dpf) (P < 0.05). At the TE level, caspase-3+ cells were frequently observed (39% = 901/2310 cells). The proportion of caspase-3+ TE cells was significantly higher in mosaic blastocysts (44.1% ± 19.6 at 5 dpf; 43% ± 16.8 at 6 dpf) and aneuploid blastocysts (45.9% ± 16.1 at 5 dpf; 49% ± 15.1 at 6 dpf) compared to euploid blastocysts (26.6% ± 16.6 at 5 dpf; 17.5% ± 14.8 at 6 dpf) (P < 0.05). In contrast, at the ICM level, caspase-3+ cells were rarely observed (1.9% = 11/596 cells), and only detected in mosaic blastocysts (2.6% = 6/232 cells) and aneuploid blastocysts (2.5% = 5/197 cells) (P > 0.05). Phase II: Consistently, TUNEL+ cells were only observed in TE cells (32.4% = 124/383 cells). An increasing trend was identified toward a higher proportion of TUNEL+ cells in the TE of mosaic blastocysts (37.2% ± 21.9) and aneuploid blastocysts (39% ± 41.7), compared to euploid blastocysts (23% ± 32.5), although these differences did not reach statistical significance (P > 0.05). LIMITATIONS, REASONS FOR CAUTION: The observed effects on apoptosis and differentiation may not be exclusive to aneuploid cells. Additionally, variations in aneuploidies and unexplored factors related to blastocyst development and karyotype concordance may introduce potential biases and uncertainties in the results. WIDER IMPLICATIONS OF THE FINDINGS: Our findings demonstrate a cell lineage-specific effect of aneuploidy on the apoptotic levels and differentiation capacity of human blastocysts. This contributes to unravelling the biological characteristics of mosaic blastocysts and supports the concept of clonal depletion of aneuploid cells in explaining their reproductive potential. STUDY FUNDING/COMPETING INTEREST(S): This work was funded by grants from Centro para el Desarrollo Tecnológico Industrial (CDTI) (20190022) and Generalitat Valenciana (APOTIP/2019/009). None of the authors has any conflict of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Probiotic Supplementation With Saccharomyces boulardii and Enterococcus faecium Improves Gastric Pain and Bloating in Airline Pilots With Chronic Non-atrophic Gastritis: An Open-Label Study.

Background Commercial airline pilots (APs) are prone to upper gastrointestinal symptoms, such as epigastric pain and bloating. These issues are often linked to occupational risk factors like irregular diet, sleep disruption, and circadian rhythm disturbance. The use of probiotics to enhance intestinal health is well established, but their efficacy in treating upper gastrointestinal diseases is still debated. This is primarily due to the stomach's small resident microbiota and its low pH, which is inhospitable to most microbes. However, emerging research suggests that specific probiotic strains, such as Enterococcus faecium, can withstand acidic environments. Moreover, certain yeast species, including Saccharomyces boulardii, can survive at a low pH. Consequently, we conducted a preliminary, three-arm, randomized, open-label, dose-finding, four-week study to compare the effects of watchful waiting (WW) with the administration of an oral probiotic supplement containing S. boulardii and E. faecium in APs diagnosed with Helicobacter pylori-negative chronic non-atrophic gastritis (CNG). Methods The study included 39 APs with CNG who were randomized into three groups with a 1:1:1 ratio. The low-dose group (n = 13) received one capsule of the probiotic supplement twice daily, before meals, for four weeks. The high-dose group (n = 13) was administered two capsules of the supplement on the same schedule. The third group (n = 13) underwent WW and served as the control arm. Blinding was maintained for the examining physicians and laboratory staff, but not for the patients. All participants self-rated their experiences of gastric pain and bloating at the beginning and conclusion of the four-week treatment period. Additionally, serum levels of pepsinogen I (PGI) and pepsinogen II (PGII) were measured at these time points. Results Supplementation with probiotics significantly outperformed WW in reducing subjective gastric pain and bloating. This effect was consistent across both tested dosages, with no significant differences observed. However, only high-dose probiotics led to a statistically significant decrease in PGII levels and an increase in the PGI/PGII ratio after the four-week study period, a result not observed with low-dose probiotics. Conclusions Oral administration of S. boulardii and E. faecium demonstrated potential efficacy in reducing gastric pain and bloating symptoms in APs with CNG, as evidenced by statistically significant symptom improvement compared to the control group that did not receive the probiotic supplementation. Notably, high-dose probiotics resulted in a significant increase in the PGI/PGII ratio, indicating potential long-term cytoprotective effects on the gastric mucosa.

Prevalence of hepatic steatosis and fibrosis in apparently healthy airline pilots: A transient elastography study.

Background and Aim: Airline pilots (APs) are often characterized by a sedentary lifestyle, predisposing them to adverse cardiometabolic consequences. In this cross-sectional study, we used transient elastography (TE) to investigate the prevalence of hepatic steatosis and fibrosis among apparently healthy APs. Materials and Methods: The study cohort consisted of 137 male APs of Caucasian descent who voluntarily underwent TE. To evaluate the extent and severity of hepatic steatosis and fibrosis, we employed established cutoff values for the controlled attenuation parameter (CAP) and liver stiffness measurement (LSM). Results: Of the APs, 34 (24.8%) were diagnosed with TE-defined steatosis. Specifically, 25 APs (18.2%) exhibited mild steatosis, 6 (4.4%) moderate steatosis, and 3 (2.2%) severe steatosis. The majority of participants (80 APs or 58.4%) showed no signs of liver fibrosis based on LSM values. However, 49 APs (35.8%) were diagnosed with mild fibrosis (F1), 7 (5.1%) with significant fibrosis (F2), and one (0.7%) with advanced fibrosis (F3). None of the pilots had F4 (cirrhosis). In multivariable linear regression analysis, BMI was the sole independent predictor of both CAP (ß=0.34, p<0.001) and LSM (ß=0.41, p<0.001) values in our sample of male APs. Conclusion: TE is a straightforward and convenient non-invasive method for detecting hepatic steatosis and fibrosis in high-risk occupational groups such as APs.

Simulations of Multifocal Vision in Patients With Previous Monofocal Intraocular Lens Implantation.

PURPOSE: To evaluate the effectiveness and patient acceptance of multifocal vision simulation in patients with previous monofocal intraocular lens (IOL) implantation, and to explore their willingness-to-pay (WTP) and willingness-to-accept (WTA) based on the perceived advantages and disadvantages of multifocal vision. METHODS: Seventeen patients with previous monofocal IOL implantation participated in this cross-sectional study. The SimVis Gekko device (2EyesVision SL) was used to simulate monofocal (Evaluation B) and multifocal (Evaluation C) visual experiences, compared to their existing vision (Evaluation A). Visual acuity at three distances and defocus curves were measured. Patients responded to inquiries about visual quality in each evaluation, bothersomeness of photic phenomena, probability to select the visual experience, and the monetary value they associated with enhanced WTP or diminished WTA visual quality. RESULTS: The simulations underestimated the visual acuity reported for the IOL in existing literature by one or two lines, depending on the testing distance. This underestimation was more pronounced in defocus curves. However, 70.6% of patients were likely or very likely to opt for multifocal vision, indicating they perceived the benefits of multifocality. The WTP for multifocal vision was twice that of monofocal vision, and the WTP/WTA ratio exceeded 1, suggesting the perceived vision benefits outweighed potential drawbacks. CONCLUSIONS: Despite underestimating the expected postoperative visual performance, the multifocal simulation enabled patients to perceive the benefits of multifocal vision to some extent. This system could be beneficial in avoiding potential postoperative complaints, but the possible rise in false-positive results should be considered and evaluated in future research. [J Refract Surg. 2023;39(12):831-839.].

Topically Applied Magnetized Saline Water Activates Autophagy in the Scalp and Increases Hair Count and Hair Mass Index in Men With Mild-to-Moderate Androgenetic Alopecia.

Introduction Water is essential for life and is vital for almost all functions of the human body. Recent studies have shown that treating water with magnets can alter its physicochemical properties, including intracluster bonds and water-ion interactions. Magnetized water also undergoes modifications in its physicochemical characteristics, such as pH, salinity, and dissolved oxygen. While there is a significant amount of literature on the use of magnetized water in agricultural settings, research on its potential biomedical applications is still limited. Based on previous findings indicating a potential relationship between autophagy activation and hair loss reversal, a pilot study was designed to explore the effects of topically applied magnetized saline water in patients with androgenetic alopecia. The hypothesis was that the process of water magnetization, which promotes the creation of hydroxyl ions, could potentially induce hair growth through the induction of alkali-induced autophagy in the scalp. Methods We recruited 20 Caucasian men with mild-to-moderate androgenetic alopecia (Norwood-Hamilton stages II-III). Initially, we conducted a 12-week open-label study to evaluate the potential of a topical lotion containing 95% magnetized saline water (2 mL applied once daily) to increase hair count and hair mass index (HMI). Subsequently, we investigated the effect of the lotion on two autophagy markers (Beclin-1 and LC3B) in scalp biopsies from a subgroup of 10 men. Results Hair count significantly increased after 12 weeks of topical treatment with magnetized saline water (from 20.6 ± 9.8 at baseline to 32.5 ± 12.4 at 12 weeks, P < 0.001). Similarly, the mean HMI increased from 37.8 ± 11.4 at baseline to 45.1 ± 13.6 at 12 weeks (P < 0.01). At the molecular level, the topical lotion effectively increased Beclin-1 levels in scalp biopsies by 44% at 12 weeks as compared to the baseline. Similarly, LC3B levels increased by 36% from baseline to 12 weeks, indicating that the lotion effectively activated autophagy in the scalp. Conclusions After 12 weeks of topical treatment, a lotion containing magnetized saline water activated scalp autophagy and significantly increased hair count and HMI in men with mild-to-moderate androgenetic alopecia.

Topically Applied Magnetized Saline Water Improves Skin Biophysical Parameters Through Autophagy Activation: A Pilot Study.

Background Water exposed to a magnetic field exhibits several changes in its properties, such as increased electrical conductivity, reduced density, and low surface tension. Additionally, it has reduced dissolved oxygen levels and becomes more alkaline. Previous experimental studies have demonstrated that exposure to saline alkaline water leads to a dose-dependent increase in the expression of autophagy-related genes. Here, we hypothesize that the topical application of magnetized alkaline water to the skin can activate autophagy and improve cutaneous biophysical parameters, making it a promising strategy for enhancing skin aesthetics. Methods Two distinct substudies were undertaken. Firstly, a 12-week, uncontrolled, open-label investigation was conducted with 20 females who desired to enhance the appearance of their facial and neck skin. Secondly, a molecular study was carried out on a subset of 10 females to investigate the serum's impact on two autophagy markers (Beclin-1 and mammalian/mechanistic target of rapamycin {mTOR}) in skin biopsies taken from the posterior neck area below the hair attachment line. Results After a period of 12 weeks, the application of the serum resulted in significant improvements in skin hydration within the stratum corneum (56 ± 14 arbitrary units {a.u.}) compared to the baseline measurement (47 ± 12 a.u.; p < 0.001). Moreover, the transepidermal water loss (TEWL) decreased from 14 ± 2 g/m2/hour to 11 ± 3 g/m2/hour (p < 0.001). The results also revealed a notable reduction in sebum content from 38 ± 7 µg/cm2 to 30 ± 4 µg/cm2 after the 12-week period of serum application (<0.001). Additionally, the melanin index (p < 0.01) and erythema index (p < 0.001) were both significantly lower at 12 weeks compared to baseline. The molecular study showed a 38% increase in Beclin-1 levels after 12 weeks of serum application on the posterior neck area, as measured from skin biopsies. In contrast, mTOR levels decreased by 24% from baseline to 12 weeks. Conclusion The application of magnetized saline water topically, within a serum formulation, shows potential in improving skin biophysical parameters for females seeking to enhance the appearance of their facial and neck skin. These beneficial effects are achieved through the activation of cutaneous autophagy, as evidenced by an increase in Beclin-1 expression and a decrease in mTOR content in the skin.