Microbial dysbiosis in roots and rhizosphere of grapevines experiencing decline is associated with active metabolic functions.

When grapevine decline, characterized by a premature decrease in vigor and yield and sometimes plant death, cannot be explained by pathological or physiological diseases, one may inquire whether the microbiological status of the soil is responsible. Previous studies have shown that the composition and structure of bacterial and fungal microbial communities in inter-row soil are affected in areas displaying vine decline, compared to areas with non-declining vines within the same plot. A more comprehensive analysis was conducted in one such plot. Although soil chemical parameters could not directly explain these differences, the declining vines presented lower vigor, yield, berry quality, and petiole mineral content than those in non-declining vines. The bacterial and fungal microbiome of the root endosphere, rhizosphere, and different horizons of the bulk soil were explored through enzymatic, metabolic diversity, and metabarcoding analysis in both areas. Despite the lower microbial diversity and richness in symptomatic roots and soil, higher microbial activity and enrichment of potentially both beneficial bacteria and pathogenic fungi were found in the declining area. Path modeling analysis linked the root microbial activity to berry quality, suggesting a determinant role of root microbiome in the berry mineral content. Furthermore, certain fungal and bacterial taxa were correlated with predicted metabolic pathways and metabolic processes assessed with Eco-Plates. These results unexpectedly revealed active microbial profiles in the belowground compartments associated with stressed vines, highlighting the interest of exploring the functional microbiota of plants, and more specifically roots and rhizosphere, under stressed conditions.

Paternal Age Amplifies Cryopreservation-Induced Stress in Human Spermatozoa.

The global fall in male fertility is a complicated process driven by a variety of factors, including environmental exposure, lifestyle, obesity, stress, and aging. The availability of assisted reproductive technology (ART) has allowed older couples to conceive, increasing the average paternal age at first childbirth. Advanced paternal age (APA), most often considered male age ≥40, has been described to impact several aspects of male reproductive physiology. In this prospective cohort study including 200 normozoospermic patients, 105 of whom were ≤35 years (non-APA), and 95 of whom were ≥42 years (APA), we assessed the impact of paternal age on different endpoints representative of sperm quality and cryopreservation tolerance. Non-APA patients had superior fresh semen quality; DNA fragmentation was notably increased in APA as compared to non-APA individuals (21.7% vs. 15.4%). Cryopreservation further increased the DNA fragmentation index in APA (26.7%) but not in non-APA patients. Additionally, APA was associated with increased mtDNAcn in both fresh and frozen/thawed sperm, which is indicative of poorer mitochondrial quality. Cryopreservation negatively impacted acrosome integrity in both age groups, as indicated by reduced incidences of unreacted acrosome in relation to fresh counterparts in non-APA (from 71.5% to 57.7%) and APA patients (from 75% to 63%). Finally, cryopreservation significantly reduced the phosphorylation status of proteins containing tyrosine residues in sperm from young males. Therefore, the present findings shed light on the effects of paternal age and cryopreservation on sperm quality and serve as valuable new parameters to improve our understanding of the mechanisms underlying sperm developmental competence that are under threat in current ART practice.

A Novel Electronic Record System for Documentation and Efficient Workflow for Community Health Workers: Development and Usability Study.

BACKGROUND: The COVID-19 pandemic added to the decades of evidence that public health institutions are routinely stretched beyond their capacity. Community health workers (CHWs) can be a crucial extension of public health resources to address health inequities, but systems to document CHW efforts are often fragmented and prone to unneeded redundancy, errors, and inefficiency. OBJECTIVE: We sought to develop a more efficient data collection system for recording the wide range of community-based efforts performed by CHWs. METHODS: The Communities Organizing to Promote Equity (COPE) project is an initiative to address health disparities across Kansas, in part, through the deployment of CHWs. Our team iteratively designed and refined the features of a novel data collection system for CHWs. Pilot tests with CHWs occurred over several months to ensure that the functionality supported their daily use. Following implementation of the database, procedures were set to sustain the collection of feedback from CHWs, community partners, and organizations with similar systems to continually modify the database to meet the needs of users. A continuous quality improvement process was conducted monthly to evaluate CHW performance; feedback was exchanged at team and individual levels regarding the continuous quality improvement results and opportunities for improvement. Further, a 15-item feedback survey was distributed to all 33 COPE CHWs and supervisors for assessing the feasibility of database features, accessibility, and overall satisfaction. RESULTS: At launch, the database had 60 active users in 20 counties. Documented client interactions begin with needs assessments (modified versions of the Arizona Self-sufficiency Matrix and PRAPARE [Protocol for Responding to and Assessing Patient Assets, Risks, and Experiences]) and continue with the longitudinal tracking of progress toward goals. A user-specific automated alerts-based dashboard displays clients needing follow-up and upcoming events. The database contains over 55,000 documented encounters across more than 5079 clients. Available resources from over 2500 community organizations have been documented. Survey data indicated that 84% (27/32) of the respondents considered the overall navigation of the database as very easy. The majority of the respondents indicated they were overall very satisfied (14/32, 44%) or satisfied (15/32, 48%) with the database. Open-ended responses indicated the database features, documentation of community organizations and visual confirmation of consent form and data storage on a Health Insurance Portability and Accountability Act-compliant record system, improved client engagement, enrollment processes, and identification of resources. CONCLUSIONS: Our database extends beyond conventional electronic medical records and provides flexibility for ever-changing needs. The COPE database provides real-world data on CHW accomplishments, thereby improving the uniformity of data collection to enhance monitoring and evaluation. This database can serve as a model for community-based documentation systems and be adapted for use in other community settings.

Exploring fibroblast interactions on nanocrystalline surfaces in physiological environments through a phenomenological lens.

The cytological behaviour and functional dynamics (adhesion, spreading, synthesis of proteins) of fibroblasts when interacting with biomedical surfaces are intricately influenced by the inherent nature of surface (nanocrystalline or microcrystalline), where the nanocrystalline (NC) surface is preferred in relation to the microcrystalline (MC) surface. This preference is a direct consequence of the distinct differences in physical and chemical characteristics between NC and MC surfaces, which include crystal boundary bio-physical attributes, electron work function, surface energy, and charge carrier density. The observed variances in cytological behaviour at the interfaces of NC and MC bio-surfaces can be attributed to these fundamental differences, particularly accounting for the percentage and nature of crystal boundaries. Recognising and understanding these physical and chemical characteristics establish the groundwork for formulating precise guidelines crucial in the development of the forthcoming generation of biomedical devices.

The significance of nanoscale surface in favourably modulating the cellular functionality is described with the aim to provide the solution to the current day challenges in the biomedical arena. Furthermore, the perspective presented advances the nano-bio science forward by implying that the nanoscale structure induces chemical and physical changes that can be considered responsible for favourable modulation of cellular activity in the living organism.

Exploring the Effects of Major Depressive Disorder on Daily Occupations and the Impact of Psychotherapy: A Literature Review.

Major depressive disorder (MDD) is a prevalent psychological mood disorder that can disrupt one's functioning and result in decreased engagement in daily activities. Psychotherapy, in different approaches, is a common approach for individuals experiencing MDD. Nevertheless, a literature review of the research supporting the effectiveness of psychotherapeutic interventions in patients with MDD-impacted areas of their daily occupations, such as back to work, cognitive deficits, and well-being, has not been conducted. A literature review was carried out to evaluate the effectiveness of psychotherapy on daily occupations for individuals diagnosed with MDD. Due to variations in study design and outcome measures, a best evidence synthesis was carried out instead of a meta-analysis. Forty-one identified articles were fully assessed in total. These studies were conducted in various countries so that a global approach could be considered comprehensive. The findings showed strong evidence supporting the effectiveness of psychotherapy on return-to-work interventions in improving depressive symptoms. There was limited evidence for the effectiveness of psychotherapy on lifestyle interventions in reducing anxiety and suicidal ideation, as well as limited evidence for enhancing work participation. Notably, there were no studies evaluating individualized client-centered psychotherapy interactions with occupations, revealing a research gap. Challenges such as incomplete reporting within studies and study heterogeneity prevented a meta-analysis. While the overall evidence base for the effectiveness of psychotherapy for MDD in treating functionality is limited, the findings provide strong support for the efficacy of occupational therapy return-to-work interventions. This is particularly important given the economic costs associated with mental health issues and work-related absences. Further research is required to strengthen the existing evidence base.

Predictive Factors of Functional Outcomes and Quality of Life in Patients with Ankle Fractures: A Systematic Review.

Background: Evaluating the predictors of unfavorable outcomes in patients with ankle fractures is crucial for identifying high-risk patients and implementing personalized treatment strategies. This study aimed to analyze factors that influence quality of life in patients with ankle fractures. Methods: Four databases were consulted. The main outcomes were functionality and quality of life scales combined using the standard mean difference (SMD) (Review Manager 5.4). Results: Eight studies with 2486 patients were included. A significant correlation was found between female sex and worse functionality scores (beta 4.15, 95% CI 1.84-6.46). Additionally, older age was correlated with worse functionality scores (beta -0.24, 95% CI -0.29 to -0.19). Patients with diabetes or metabolic syndrome also had worse outcomes (SMD 0.27, 95% CI 0.18-0.36). High BMI and obesity were also associated with worse quality of life scores (beta 2.62, 95% CI 0.77-4.48). Smokers had greater disability in the analyzed scales (SMD 0.22, 95% CI 0.05-0.39). No significant differences were observed with respect to syndesmotic involvement. Conclusions: Age, sex, diabetes, high BMI, and smoking negatively impact functional outcomes and quality of life in patients with ankle fractures.

Functional Outcomes in Upper Limb Replantation-A Systematic Review.

Functionality after upper limb replantation is a decisive factor when considering the success of the intervention. Therefore, its evaluation is fundamental. The aim of this article was to conduct a systematic review of upper limb functions after replantation or reimplantation, seeking to identify reported functional outcomes as well as the level and mechanism of injury. To achieve this objective, a literature search was conducted in PubMed, ScienceDirect, Cochrane and Web of Science. Studies from the last 10 years which included patients with upper limb replantation and reported their functional outcomes were included. Out of 523 articles, 12 studies (n = 607) were finally included. DASH and CISS were the most commonly used assessments to report functional outcomes. In conclusion, functional outcomes after replantation are assessed using widely varying scales; therefore, due to this methodological variability, it is difficult to compare functional success between studies and further studies on functionality are needed to provide new data.

Long-term Success of Regenerative Endodontic Procedures.

Background: Regenerative endodontic procedures (REPs) have emerged as a transformative approach to treating immature permanent teeth with necrotic pulp tissue. Materials and Methods: A prospective study was conducted, enrolling 100 patients with immature permanent teeth requiring REPs. All procedures were performed by a single experienced endodontist following established protocols. Patients were followed up for a minimum of 5 years' post-treatment. Clinical examinations, radiographic assessments, and patient-reported outcomes were recorded at regular intervals. Data were analyzed using statistical methods to determine the success rates, complications, and factors influencing long-term outcomes. Results: The results of this original research reveal a significant and sustained success rate for REPs. After a minimum follow-up period of 5 years, an arbitrary value of 92% for tooth survival was achieved. Radiographic assessments demonstrated consistent healing of apical lesions, and continued root development was observed in the majority of cases. Patient-reported outcomes indicated a high level of satisfaction with the procedure. Complications such as crown discoloration and tooth fracture occurred in a minority of cases but were effectively managed without compromising the overall success of REPs. Conclusion: This original research provides strong evidence for the long-term success of REPs in the treatment of immature permanent teeth with necrotic pulp tissue. The high tooth survival rate, continued root development, and patient satisfaction support the efficacy of REPs as a reliable treatment option.

Effects of combination of strength and balance training on postural control and functionality in people with chronic ankle instability: a systematic review and meta analysis.

AIM: To identify the effects of strength and balance training on dynamic balance and patient reported outcomes in people with chronic ankle instability(CAI). METHOD: Five databases(CNKI, WanFang, Web of Science, EBSCO-SPORTD and PubMed were searched in September 2022. The search was conducted on randomized controlled trials(RCTs) that the effects of strength training, balance training and combination of strength and balance training in people with chronic ankle instability compared to a control group. Using Review Manager 5.3 and Stata-SE 15 to conduct Meta-analysis on the included literature. methodological quality and risk of bias were assessed by using the PEDro scale. RESULTS: A total of 33 Chinese and English RCTs document were screened and 1154 patients with CAI were included in the study. Compared with control group, strength training, balance training and combination of strength and balance training demonstrated to be more effective in terms of improving patient reported outcomes(strength training: SMD = 0.80, 95%CI = 0.39-1.22; balance training: SMD = 0.79, 95%CI = 0.41-1.17; combination of strength and balance training: SMD = 1.28, 95%CI = 0.57, 1.99). Subgroup analysis: Intervention for 6 weeks, more than 3 times a week and more than 30 min each time were the best rehabilitation programs to improve CAI patientreported outcomes. Compared with control group, balance training demonstrated to be more effective in terms of improving Star Excursion Balance Test (SEBT)((anterior: SMD = 0.71, 95%CI = 0.03-1.40; posterolateral: SMD = 0.84, 95%CI = 0.22-1.46; posteromedial: SMD = 0.88, 95%CI = 0.45-1.32). However, strength training and combination of strength and balance training had no improvement effects on SEBT. CONCLUSIONS: Available evidence showed that, results of the comparison between balance training versus strength training suggest that the combination of strength and balance training achieves greater benefits for patient reported outcomes and balance training could bring greater benefits to dynamic balance. Strength training should be used cautiously in clinic to improve the dynamic balance in individuals with CAI. TRIAL REGISTRATION: ( http://www.crd.york.ac.uk/PROSPERO , Registration No. CRD42022371396).

Asociación entre funcionalidad familiar y deterioro cognitivo leve en la familia con adulto mayor / Association between family functionality and mild cognitive impairment in the family with the elderly

Objetivo: Determinar la asociación entre funcionalidad familiar y deterioro cognitivo leve en la familia con adulto mayor. Metodología: Diseño transversal analítico en familias con pacientes geriátricos. Se consideró familia con paciente geriátrico cuando al menos uno de sus integrantes tenía más de 60años de edad. Los grupos de comparación fueron la familia con paciente geriátrico sin deterioro cognitivo y la familia con paciente geriátrico con deterioro cognitivo leve determinada con el instrumento MoCA. La funcionalidad familiar se evaluó con el instrumento APGAR familiar, el cual identifica tres categorías: funcionalidad familiar, disfuncionalidad familiar moderada y disfuncionalidad familiar severa. El análisis estadístico incluyó chi cuadrado y prueba de Mann Whitney. Resultados: En la familia con paciente geriátrico, en el grupo sin deterioro cognitivo la prevalencia de funcionalidad familiar es del 89,7% y en el grupo con deterioro cognitivo leve la prevalencia de funcionalidad familiar es del 59,3% (MW=4,87, p<0,000). Conclusión: Existe asociación entre funcionalidad familiar y deterioro cognitivo leve.(AU)

Aim: To determine the association between family functionality and mild cognitive impairment in the family with the elderly. Methodology: Analytical cross-sectional design in families with geriatric patients. A family with a geriatric patient was considered when at least one of its members was over 60years of age. The comparison groups were the family with a geriatric patient without cognitive impairment and the family with a geriatric patient with mild cognitive impairment determined with the MoCA instrument. Family functionality was evaluated with the family APGAR instrument, which identifies three categories: family functionality, moderate family dysfunction, and severe family dysfunction. Statistical analysis included Chi square and Mann-Whitney test. Results: In the family with a geriatric patient, in the group without cognitive impairment the prevalence of family functionality is 89.7% and in the group with mild cognitive impairment the prevalence of family functionality is 59.3% (MW=4.87, P<.000). Conclusion: There is an association between family functionality and mild cognitive impairment.(AU)

Eficacia del protocolo Start to move en funcionalidad, DA-UCI y delirio: ensayo clínico aleatorizado / Efficacy of the «Start to move» protocol on functionality, ICU-acquired weakness and delirium: Randomized clinical trial

Objetivo Evaluar la eficacia del protocolo Start to move comparado con el tratamiento convencional en sujetos mayores de 15 años hospitalizados en la UCI sobre una mejoría en funcionalidad, disminución de debilidad adquirida en la UCI (DA-UCI), incidencia de delirio, días de ventilación mecánica (VM), estadía en la UCI y mortalidad a los 28 días. Diseño Ensayo clínico controlado aleatorizado. Ámbito Unidad de paciente crítico. Participantes Incluye adultos mayores a 15 años con VMI mayor a 48h, asignación aleatoria. Intervenciones Protocolo «Start to move» y tratamiento convencional. Variables de interés principales Se analizó funcionalidad, incidencia DA-UCI, incidencia delirio, días VM, estadía UCI y mortalidad-28 días, ClinicalTrials.gov número, NCT05053724. Resultados Sesenta y nueve sujetos fueron ingresados al estudio, 33 al grupo Start to move y 36 a tratamiento convencional, comparables clínico y sociodemograficamente. En el grupo Start to move la incidencia DAUCI al egreso de la UCI fue de 35,7 vs. 80,7% grupo tratamiento convencional (p=0,001). La funcionalidad (FSS-ICU) al egreso de la UCI corresponde a 26 vs. 17 puntos a favor del grupo Start to move (p=0,001). La diferencia en Barthel al egreso de la UCI fue del 20% a favor del grupo Start to move (p=0,006). No hubo diferencias significativas en incidencia de delirio, días de VM, estadía UCI y mortalidad-28 días. El estudio no reportó eventos adversos, ni suspensión de protocolo. Conclusiones La aplicación del protocolo Start to move en la UCI se asoció reducción en la incidencia DA-UCI, aumento en funcionalidad y menor caída en puntaje Barthel al egreso. (AU)

Objective To evaluate the efficacy of the Start to move protocol compared to conventional treatment in subjects over 15 years of age hospitalized in the ICU on an improvement in functionality, decrease in ICU-acquired weakness (IUCD), incidence of delirium, days of mechanical ventilation (MV), length of stay in ICU and mortality at 28 days. Design Randomized controlled clinical trial. Setting Intensive care unit. Participants Includes adults older than 15 years with invasive mechanical ventilation more than 48h, randomized allocation. Interventions Start to move protocol and conventional treatment. Main variables of interest Functionality, incidence of ICU-acquired weakness, incidence of delirium, days on mechanical ventilation, ICU stay and mortality-28 days, ClinicalTrials.gov number, NCT05053724. Results Sixty-nine subjects were admitted to the study, 33 to the Start to move group and 36 to conventional treatment, clinically and sociodemographic comparable. In the “Start to move” group, the incidence of IUCD at ICU discharge was 35.7% vs. 80.7% in the “conventional treatment” group (P=.001). Functionality (FSS-ICU) at ICU discharge corresponds to 26 vs. 17 points in favor of the “Start to move” group (P=.001). The difference in Barthel at ICU discharge was 20% in favor of the “Start to move” group (P=.006). There were no significant differences in the incidence of delirium, days of mechanical ventilation, ICU stay and 28-day mortality. The study did not report adverse events or protocol suspension. Conclusions The application of the “Start to move” protocol in ICU showed a reduction in the incidence of IUCD, an increase in functionality and a smaller decrease in Barthel score at discharge. (AU)

Functionality and loneliness in older people: Model of mediation by positive affect / Funcionalidad y soledad en personas mayores: Modelo de mediación por el afecto positivo

Objetivo: La soledad se asocia a la discapacidad funcional en la población mayor, aunque se sabe poco sobre el papel de la afectividad en esta relación. Nos proponemos explorar el efecto mediador de la afectividad en la relación entre funcionalidad y soledad, controlando el efecto de las variables relevantes. Métodos: Se administró la Escala de Soledad de la Universidad de California-16, la Escala de Funcionalidad Geriátrica, las Escalas de Afecto Positivo y Negativo-14 y la Escala de Depresión Geriátrica-8 a 489 adultos mayores (65–100 años), 428 de residencias geriátricas y 61 de la comunidad. Resultados: Los residentes en una institución, mujeres, viudos, con baja educación, más discapacidad funcional, más afecto negativo, menos afecto positivo y más síntomas depresivos reportaron más soledad. Controlando el efecto de los síntomas depresivos, género, situación residencial, estado civil y educación, solo el afecto positivo medió, parcialmente, la relación entre funcionalidad y soledad. Conclusión: Se sugieren intervenciones para aumentar la concienciación por parte de los adultos mayores en los perfiles personalizados de afecto positivo, aliviando así los sentimientos de soledad en los que sufren limitaciones funcionales (especialmente mujeres, con síntomas depresivos, viudos, residiendo en una institución y con bajo nivel educativo).(AU)

Objective: Research has shown that loneliness is associated with functional disability in the older population. However, little is known about the role of affectivity in this relationship. The present study explored a mediation model in which affectivity was hypothesized to mediate the re-lationship between functionality and loneliness, controlling for the effect of relevant variables.Methods:The University of California Loneliness Scale-16 items, Geriatric Functionality Scale, Positive and Negative AffectSchedule-14, and Geriatric Depression Scale-8 were administered to 489 old adults (65–100 years old), 428 from social care homes (SCH), and 61 from the community.Results:Those from SCH, women, widowed, withlow education, more functional disability, more negative affect, less posi-tive affect, and more depressive symptoms reported more loneliness. Con-trolling for the effect of depressive symptoms, gender, residential status, marital status, and education, only positive affect mediated, partially, the relationship between functionality and loneliness.Conclusion:We suggest implementing interventions that increase older adults' insights in personal-ized patterns of positive affect and, consequently, ease feelings of loneli-ness in older people suffering from functional limitations (especially wom-en, with depressive symptoms, widowed, residing in an institution, and with low educational level).(AU)

All-Inside Arthroscopic Meniscus Repair for Patients Over 40 Years of Age: Is Forty the New Twenty?

INTRODUCTION: Meniscal injuries are a common challenge in orthopedic surgery. For decades, partial or total meniscectomy has been the primary surgical treatment for meniscal tears. In recent years, the increased recognition of menisci's biomechanical importance has progressively shifted the paradigm towards meniscus repair. However, meniscus-sparing surgery remains the treatment of choice for selected lesions in young and active patients, especially for young and active patients. In this study, we evaluated the effectiveness of all-inside sutures in treating meniscus tears in patients over 40. METHODS: In our retrospective evaluation, we evaluated the clinical and functional outcomes of cases over 40 years of age with post-traumatic acute meniscus tears that received meniscus repairs using the all-inside technique. The pre-operative and post-operative functionality of treated patients were assessed using the knee injury and osteoarthritis outcome score (KOOS) score. Major complications were recorded. RESULTS: Twenty-three cases met our inclusion criteria. Eight females and fifteen males with a mean age of 44.9 were included in our study. Their mean follow-up was 35.1 months. Before surgery, our patients' mean KOOS score was 55.4 (18-80). At the patients' latest follow-up, the value had risen to 87.4 (63-100). There was no statistical correlation between patients' age and their functional recovery. No surgical failure was recorded at the latest follow-up. CONCLUSION: The all-inside suture technique can represent a suitable and reliable solution for suturable meniscal tears, even for patients over 40. Preserving the meniscus and restoring patients' functionality allows patients to return to their daily activities and promote their quality of life.

Evolution of the catalytic mechanism at the dawn of the Baeyer-Villiger monooxygenases.

Enzymes are crucial for the emergence and sustenance of life on earth. How they became catalytically active during their evolution is still an open question. Two opposite explanations are plausible: acquiring a mechanism in a series of discrete steps or all at once in a single evolutionary event. Here, we use molecular phylogeny, ancestral sequence reconstruction, and biochemical characterization to follow the evolution of a specialized group of flavoprotein monooxygenases, the bacterial Baeyer-Villiger monooxygenases (BVMOs). These enzymes catalyze an intricate chemical reaction relying on three different elements: a reduced nicotinamide cofactor, dioxygen, and a substrate. Characterization of ancestral BVMOs shows that the catalytic mechanism evolved in a series of steps starting from a FAD-binding protein and further acquiring reactivity and specificity toward each of the elements participating in the reaction. Together, the results of our work portray how an intrinsically complex catalytic mechanism emerged during evolution.

Ecology of Legionella pneumophila biofilms: The link between transcriptional activity and the biphasic cycle.

There has been considerable discussion regarding the environmental life cycle of Legionella pneumophila and its virulence potential in natural and man-made water systems. On the other hand, the bacterium's morphogenetic mechanisms within host cells (amoeba and macrophages) have been well documented and are linked to its ability to transition from a non-virulent, replicative state to an infectious, transmissive state. Although the morphogenetic mechanisms associated with the formation and detachment of the L. pneumophila biofilm have also been described, the capacity of the bacteria to multiply extracellularly is not generally accepted. However, several studies have shown genetic pathways within the biofilm that resemble intracellular mechanisms. Understanding the functionality of L. pneumophila cells within a biofilm is fundamental for assessing the ecology and evaluating how the biofilm architecture influences L. pneumophila survival and persistence in water systems. This manuscript provides an overview of the biphasic cycle of L. pneumophila and its implications in associated intracellular mechanisms in amoeba. It also examines the molecular pathways and gene regulation involved in L. pneumophila biofilm formation and dissemination. A holistic analysis of the transcriptional activities in L. pneumophila biofilms is provided, combining the information of intracellular mechanisms in a comprehensive outline. Furthermore, this review discusses the techniques that can be used to study the morphogenetic states of the bacteria within biofilms, at the single cell and population levels.

Assessing the functionality of a water-vending kiosk network with high-frequency instrumentation in Freetown, Sierra Leone.

Access to safe, reliable, and equitable water services in urban settings of low- and middle-income countries remains a critical challenge toward achieving Sustainable Development Goal 6.1, but progress has either slowed or stagnated in recent years. A pilot water kiosk network funded by the United States Millennium Challenge Corporation was implemented by the Sierra Leone Millennium Challenge Coordinating Unit into the intermittent piped water distribution network of Freetown, Sierra Leone, as a private-public partnership to improve water service provision for households without reliable piped water connections and to reduce non-revenue water. This study employs the use of high-frequency instrumentation to monitor, model, and assess the functionality of this water kiosk network over 2,947 kiosk-days. Functionality was defined via functionality levels on a daily basis through monitored stored water levels and modeled water withdrawals. The functionality levels across the kiosk network were found to be 34% operational, 30% offline, and 35% empty. Statistically significant (p<0.001) determinants of functionality were found for several predictors across the defined thresholds. Finally, modeling of water supply, water demand and withdrawal capacity, and water storage was conducted to further explain findings and provide additionally externally relevant support for kiosk operations.

From the Clinical to the Bench: Exploring the Insulin Modulation Effects of Tacrolimus and Belatacept.

Calcineurin inhibitors (CNIs) are critical in preventing rejection posttransplantation but pose an increased risk of post-transplant diabetes (PTD). Recent studies show that late conversion from CNIs to belatacept, a costimulation blocker, improves HbA1c in kidney transplant recipients with PTD or de novo diabetes. This study investigates whether the observed effects on PTD stem solely from CNI withdrawal or if belatacept influences PTD independently. The study assessed the impact of tacrolimus and belatacept on insulin secretion in MIN6 cells (a beta cell line) and rat islets. Tacrolimus and belatacept were administered to the cells and islets, followed by assessments of cell viability and insulin secretion. Tacrolimus impaired insulin secretion without affecting cell viability, while belatacept showed no detrimental effects on either parameter. These findings support clinical observations of improved HbA1c upon switching from tacrolimus to belatacept. Belatacept holds promise in islet or pancreas transplantation, particularly in patients with unstable diabetes. Successful cases of islet transplantation treated with belatacept without severe hypoglycemia highlight its potential in managing PTD. Further research is needed to fully understand the metabolic changes accompanying the transition from CNIs to belatacept. Preserving insulin secretion emerges as a promising avenue for investigation in this context.

Assessment of Connexin43 Hemichannel Functionality Based on Cytosolic Uptake of Yo-Pro1.

Connexin proteins are the building blocks of gap junctions and connexin hemichannels. Both provide a pathway for cellular communication. Gap junctions support intercellular communication mechanisms and regulate homeostasis. In contrast, open connexin hemichannels connect the intracellular compartment and the extracellular environment, and their activation fuels inflammation and cell death. The development of clinically applicable connexin hemichannel blockers for therapeutic purposes is therefore gaining momentum. This chapter describes a well-established protocol optimized for assessing connexin hemichannel activity by using the reporter dye Yo-Pro1.

Measurement of Ca2+ Uptake Through Connexin Hemichannels.

Increasing evidence points to deregulated flux of ionized calcium (Ca2+) mediated by hyperactive mutant connexin (Cx) hemichannels (HCs) as a common gain-of-function etiopathogenetic mechanism for several diseases, ranging from skin disorders to nervous system defects. Furthermore, the opening of nonmutated Cx HCs is associated with an impressive list of widespread diseases including, but not limited to, ischemia/stroke, Alzheimer's disease, and epilepsy. HC inhibitors are attracting a growing attention due to their therapeutic potential for numerous pathologies. This chapter describes a quantitative method to measure Ca2+ uptake though HCs expressed in cultured cells. The assay we developed can be used to probe HC activity as wells as to test HC inhibitors. Furthermore, with minor changes it can be easily adapted to high-throughput high-content platforms and/or primary cells and microtissues.

Role of Cytoskeletal Elements in Regulation of Synaptic Functions: Implications Toward Alzheimer's Disease and Phytochemicals-Based Interventions.

Alzheimer's disease (AD), a multifactorial disease, is characterized by the accumulation of neurofibrillary tangles (NFTs) and amyloid beta (Aß) plaques. AD is triggered via several factors like alteration in cytoskeletal proteins, a mutation in presenilin 1 (PSEN1), presenilin 2 (PSEN2), amyloid precursor protein (APP), and post-translational modifications (PTMs) in the cytoskeletal elements. Owing to the major structural and functional role of cytoskeletal elements, like the organization of axon initial segmentation, dendritic spines, synaptic regulation, and delivery of cargo at the synapse; modulation of these elements plays an important role in AD pathogenesis; like Tau is a microtubule-associated protein that stabilizes the microtubules, and it also causes inhibition of nucleo-cytoplasmic transportation by disrupting the integrity of nuclear pore complex. One of the major cytoskeletal elements, actin and its dynamics, regulate the dendritic spine structure and functions; impairments have been documented towards learning and memory defects. The second major constituent of these cytoskeletal elements, microtubules, are necessary for the delivery of the cargo, like ion channels and receptors at the synaptic membranes, whereas actin-binding protein, i.e., Cofilin's activation form rod-like structures, is involved in the formation of paired helical filaments (PHFs) observed in AD. Also, the glial cells rely on their cytoskeleton to maintain synaptic functionality. Thus, making cytoskeletal elements and their regulation in synaptic structure and function as an important aspect to be focused for better management and targeting AD pathology. This review advocates exploring phytochemicals and Ayurvedic plant extracts against AD by elucidating their neuroprotective mechanisms involving cytoskeletal modulation and enhancing synaptic plasticity. However, challenges include their limited bioavailability due to the poor solubility and the limited potential to cross the blood-brain barrier (BBB), emphasizing the need for targeted strategies to improve therapeutic efficacy.