¿Agua para todos? La intermitencia en el suministro de agua en los hogares en México.

OBJETIVO: Medir el acceso a través de la intermitencia en el suministro de agua potable en hogares mexicanos. Material y métodos. A través de la Encuesta Nacional de Salud y Nutrición 2022 (Ensanut 2022), se recolectó información sobre intermitencia en días por semana y horas por día durante las últimas cuatro semanas y el suministro de agua durante el año para la temporada de mayor escasez. RESULTADOS: 31.5% de los hogares recibieron agua los siete días de la semana, las 24 horas del día. De estos, 17.4% no tuvo escasez en los últimos 12 meses. La intermitencia es más común entre hogares de las regiones en el sur del país y entre los más pobres. El 81% de las familias almacena agua y 16% almacena en contenedores portátiles como cubetas. Conclusión. En este artículo se presentan por primera vez patrones de intermitencia en el suministro de agua a nivel nacional en México. La gran mayoría de las familias no reciben agua de forma continua y tienen que almacenar agua. El almacenamiento podría disminuir la calidad del agua y la falta de confianza para su consumo con consecuencias para la salud. La conexión al sistema potable no refleja el acceso real de las familias al agua.

Daylight photodynamic therapy as a treatment for for actinic field change in patients diagnosed with Oculocutaneous albinism in Sub-Saharan Africa.

BACKGROUND: Oculocutaneous albinism (OCA) is a genetically inherited condition, whereby melanin is reduced or absent in the skin. A lack of melanin predisposes patients to actinic damage and skin malignancies. In Tanzania, a resource-limited country, the treatment of choice for pre-cancerous skin lesions is cryotherapy. OBJECTIVES: To investigate whether daylight PDT is a safe and well-tolerated treatment for actinic field change in the OCA population in Tanzania. METHODS: 12 participants with actinic damage were recruited from a Standing Voice skin surveillance clinic and treated with dPDT. Study participants completed tolerability and acceptability questionnaires at day 5 and 3-months post-treatment. A dermatologist assessed clinical response to dPDT at 3 months. RESULTS: Daylight PDT was well-tolerated and acceptable to the majority of patients. Actinic damage was reduced by 25-90%. No skin cancers developed during the treatment. CONCLUSIONS: This pilot study suggests that dPDT is a safe and tolerable treatment for actinic damage in the OCA population in Tanzania. Further work is required to compare the efficacy of dPDT against other topical therapies for actinic field change.

The Frequency of Symptoms in Patients With a Diagnosis of Degenerative Cervical Myelopathy: Results of a Scoping Review.

STUDY DESIGN: Delayed diagnosis of degenerative cervical myelopathy (DCM) is associated with reduced quality of life and greater disability. Developing diagnostic criteria for DCM has been identified as a top research priority. OBJECTIVES: This scoping review aims to address the following questions: What is the diagnostic accuracy and frequency of clinical symptoms in patients with DCM? METHODS: A scoping review was conducted using a database of all primary DCM studies published between 2005 and 2020. Studies were included if they (i) assessed the diagnostic accuracy of a symptom using an appropriate control group or (ii) reported the frequency of a symptom in a cohort of DCM patients. RESULTS: This review identified three studies that discussed the diagnostic accuracy of various symptoms and included a control group. An additional 58 reported on the frequency of symptoms in a cohort of patients with DCM. The most frequent and sensitive symptoms in DCM include unspecified paresthesias (86%), hand numbness (82%) and hand paresthesias (79%). Neck and/or shoulder pain was present in 51% of patients with DCM, whereas a minority had back (19%) or lower extremity pain (10%). Bladder dysfunction was uncommon (38%) although more frequent than bowel (23%) and sexual impairment (4%). Gait impairment is also commonly seen in patients with DCM (72%). CONCLUSION: Patients with DCM present with many different symptoms, most commonly sensorimotor impairment of the upper extremities, pain, bladder dysfunction and gait disturbance. If patients present with a combination of these symptoms, further neuroimaging is indicated to confirm the diagnosis of DCM.

The value of Clinical signs in the diagnosis of Degenerative Cervical Myelopathy - A Systematic review and Meta-analysis.

STUDY DESIGN: Delayed diagnosis of degenerative cervical myelopathy (DCM) is likely due to a combination of its subtle symptoms, incomplete neurological assessments by clinicians and a lack of public and professional awareness. Diagnostic criteria for DCM will likely facilitate earlier referral for definitive management. OBJECTIVES: This systematic review aims to determine (i) the diagnostic accuracy of various clinical signs and (ii) the association between clinical signs and disease severity in DCM? METHODS: A search was performed to identify studies on adult patients that evaluated the diagnostic accuracy of a clinical sign used for diagnosing DCM. Studies were also included if they assessed the association between the presence of a clinical sign and disease severity. The QUADAS-2 tool was used to evaluate the risk of bias of individual studies. RESULTS: This review identified eleven studies that used a control group to evaluate the diagnostic accuracy of various signs. An additional 61 articles reported on the frequency of clinical signs in a cohort of DCM patients. The most sensitive clinical tests for diagnosing DCM were the Tromner and hyperreflexia, whereas the most specific tests were the Babinski, Tromner, clonus and inverted supinator sign. Five studies evaluated the association between the presence of various clinical signs and disease severity. There was no definite association between Hoffmann sign, Babinski sign or hyperreflexia and disease severity. CONCLUSION: The presence of clinical signs suggesting spinal cord compression should encourage health care professionals to pursue further investigation, such as neuroimaging to either confirm or refute a diagnosis of DCM.

Toward Shared Decision-Making in Degenerative Cervical Myelopathy: Protocol for a Mixed Methods Study.

BACKGROUND: Health care decisions are a critical determinant in the evolution of chronic illness. In shared decision-making (SDM), patients and clinicians work collaboratively to reach evidence-based health decisions that align with individual circumstances, values, and preferences. This personalized approach to clinical care likely has substantial benefits in the oversight of degenerative cervical myelopathy (DCM), a type of nontraumatic spinal cord injury. Its chronicity, heterogeneous clinical presentation, complex management, and variable disease course engenders an imperative for a patient-centric approach that accounts for each patient's unique needs and priorities. Inadequate patient knowledge about the condition and an incomplete understanding of the critical decision points that arise during the course of care currently hinder the fruitful participation of health care providers and patients in SDM. This study protocol presents the rationale for deploying SDM for DCM and delineates the groundwork required to achieve this. OBJECTIVE: The study's primary outcome is the development of a comprehensive checklist to be implemented upon diagnosis that provides patients with essential information necessary to support their informed decision-making. This is known as a core information set (CIS). The secondary outcome is the creation of a detailed process map that provides a diagrammatic representation of the global care workflows and cognitive processes involved in DCM care. Characterizing the critical decision points along a patient's journey will allow for an effective exploration of SDM tools for routine clinical practice to enhance patient-centered care and improve clinical outcomes. METHODS: Both CISs and process maps are coproduced iteratively through a collaborative process involving the input and consensus of key stakeholders. This will be facilitated by Myelopathy.org, a global DCM charity, through its Research Objectives and Common Data Elements for Degenerative Cervical Myelopathy community. To develop the CIS, a 3-round, web-based Delphi process will be used, starting with a baseline list of information items derived from a recent scoping review of educational materials in DCM, patient interviews, and a qualitative survey of professionals. A priori criteria for achieving consensus are specified. The process map will be developed iteratively using semistructured interviews with patients and professionals and validated by key stakeholders. RESULTS: Recruitment for the Delphi consensus study began in April 2023. The pilot-testing of process map interview participants started simultaneously, with the formulation of an initial baseline map underway. CONCLUSIONS: This protocol marks the first attempt to provide a starting point for investigating SDM in DCM. The primary work centers on developing an educational tool for use in diagnosis to enable enhanced onward decision-making. The wider objective is to aid stakeholders in developing SDM tools by identifying critical decision junctures in DCM care. Through these approaches, we aim to provide an exhaustive launchpad for formulating SDM tools in the wider DCM community. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/46809.

Preventative Management of Sepsis-Induced Acute Respiratory Distress Syndrome in the Geriatric Population.

Sepsis and its treatment are the most common etiologies of acute respiratory distress syndrome (ARDS), which has a disturbing mortality rate. Sepsis management relies heavily on the introduction of resuscitative fluids. However, when fluids are paired with the circulating inflammatory mediators of sepsis, patients are prone to lung damage. Survivors of sepsis-induced ARDS become plagued with functional and/or psychological sequelae such as impaired memory, difficulty in concentrating, and decreased mental processing speed. Specific techniques can be implemented when diagnosing and treating elderly patients with sepsis to prevent the onset of ARDS, including bed elevation and early antibiotics. Additionally, albumin infusion may be beneficial; however, more research must be conducted. Finally, inflammatory mediators, including serum mannose biomarkers and extracellular histone therapy, show a promising avenue for future treatment. Although there is limited research on osteopathic manipulative medicine (OMT) on ARDS or sepsis-induced ARDS, OMT that focuses on alleviating rib and thoracic somatic dysfunctions has been used as an adjunct therapy to treat other respiratory diseases, such as pneumonia and chronic obstructive pulmonary disease (COPD). The results of these studies may garner interest in whether the use of OMT as an adjunct therapy may be beneficial for patients with ARDS or sepsis-induced ARDS. This paper is intended to review the current guidelines for sepsis and ARDS management in elderly patients to identify measures to prevent sepsis-induced ARDS.

Clinical and epidemiologic features of persons accessing emergency departments for dog and cat bite injuries in California (2005-2019).

OBJECTIVE: To describe the clinical and epidemiologic features of persons with dog and cat bite injuries who presented to emergency departments. SAMPLE: Records of 648,492 dog and cat bite-related emergency department visits in California from 2005 to 2019. PROCEDURES: Visits were selected by standardized International Classification of Diseases, Ninth or Tenth Revision, Clinical Modification codes that indicated a bite as an external cause of injury in the medical record. Incidence rates were calculated for patient demographics, location and month of bite incident, characteristics of bite injury, infection, patient outcome/disposition, and expected source of payment. Cross-sectional descriptive analysis was performed. RESULTS: The average annual incidence of dog bites was highest in children aged < 10 years and males, while that of cat bites was highest in adults aged ≥ 80 years and females. Bites were more likely to occur in rural settings, in private residences, and during the summer. The median household income for zip codes in which animal bite patients resided was lower than the statewide median household income. Both dog and cat bite injuries were more likely to occur to upper limbs. Bacteria were isolated from 3% of dog bite injuries and 21.5% of cat bite injuries at initial presentation. CLINICAL RELEVANCE: Epidemiologic findings about persons presenting to emergency departments for animal bite injuries can inform bite prevention efforts by identifying at-risk populations. Effective animal bite prevention demands an ongoing multisectoral program of veterinarians and other health professionals, collaborating with community and governmental organizations, to develop and implement integrated strategies within the context of other socially contributory factors.

"We're Constantly Learning": Identifying and Disrupting White Supremacy Within a Victim Services Organization.

Many victim service organizations are seeking to realign service delivery around principles of racial equity. Dismantling institutional racism is a complex, intensive, and long-term process. Therefore, despite this imperative from the field, our knowledge about how social service organizations can effectively advance anti-oppressive practice is limited. This study examined victim advocate perspectives on the role institutional racism played within their work and the supports needed to undo institutional racism within their organization. Six focus groups were conducted with a meaningful cross section of staff members (n = 53) across the organization. Semi-structured interview guides included questions in four domains: (1) racism within client work, (2) challenges to addressing racism, (3) effective solutions, and (4) helpful organizational supports. Transcripts were thematically analyzed using modified constructivist grounded theory methods. Two overarching themes, Identifying Institutional Racism in the Workplace and Advancing Anti-racist Practice, and six subthemes emerged from the analysis. Advocates identified that naming and becoming comfortable talking about race was essential. Further, they believed it was important to acknowledge the ways in which that racism was implicitly built into helping systems at large. Advocates explored how internalized racial stereotypes influenced interactions between black, indigenous, and people of color (BIPOC) and white advocates and their clients in complex ways. Advocates highlighted organizational efforts that supported ongoing personal reflection, the creation of an accountable community, and staff empowerment within the organization as being critical to advancing anti-racist practice. Some advocates also wanted to see the organization move further in the direction of standing with BIPOC communities, particularly around criminal justice concerns. Findings provide important timely insights into how institutional racism manifests within victim service organizations and what organizational actions encourage anti-oppressive practices and culture.

Disseminated Intravascular Coagulation in Varying Age Groups Based on Clinical Conditions.

Disseminated intravascular coagulation (DIC) is a serious syndrome characterized by the systemic activation of blood coagulation resulting in the thrombosis of vessels leading to organ dysfunction and severe bleeding. When physicians try to treat DIC, it is imperative to diagnose and treat the underlying conditions. Anyone can be affected by DIC, but vulnerable groups such as pediatric populations, pregnant women and the elderly may be at higher risk. In this review, the current literature on DIC in pregnancy, the pediatric population, and the elderly is reported. This review also highlights the similarities and differences in the etiology, clinical presentation, diagnosis, and management of DIC in the aforementioned groups (i.e., pediatrics, pregnant women, and the elderly). Findings from this study may help increase awareness about various presentations of DIC in these groups to facilitate rapid recognition of symptoms leading to correct diagnoses.

Improving patient-centred care through a tailored intervention addressing nursing clinical handover communication in its organizational and cultural context.

AIMS: To increase the quality and safety of patient care, many hospitals have mandated that nursing clinical handover occur at the patient's bedside. This study aims to improve the patient-centredness of nursing handover by addressing the communication challenges of bedside handover and the organizational and cultural practices that shape handover. DESIGN: Qualitative linguistic ethnographic design combining discourse analysis of actual handover interactions and interviews and focus groups before and after a tailored intervention. METHODS: Pre-intervention we conducted interviews with nursing, medical and allied health staff (n = 14) and focus groups with nurses and students (n = 13) in one hospital's Rehabilitation ward. We recorded handovers (n = 16) and multidisciplinary team huddles (n = 3). An intervention of communication training and recommendations for organizational and cultural change was delivered to staff and championed by ward management. After the intervention we interviewed nurses and recorded and analyzed handovers. Data were collected from February to August 2020. Ward management collected hospital-acquired complication data. RESULTS: Notable changes post-intervention included a shift to involve patients in bedside handovers, improved ward-level communication and culture, and an associated decrease in reported hospital-acquired complications. CONCLUSIONS: Effective change in handover practices is achieved through communication training combined with redesign of local practices inhibiting patient-centred handovers. Strong leadership to champion change, ongoing mentoring and reinforcement of new practices, and collaboration with nurses throughout the change process were critical to success. IMPACT: Ineffective communication during handover jeopardizes patient safety and limits patient involvement. Our targeted, locally designed communication intervention significantly improved handover practices and patient involvement through the use of informational and interactional protocols, and redesigned handover tools and meetings. Our approach promoted a ward culture that prioritizes patient-centred care and patient safety. This innovative intervention resulted in an associated decrease in hospital-acquired complications. The intervention has been rolled out to a further five wards across two hospitals.

Sleep Difficulties among Mexican Adolescents: Subjective and Objective Assessments of Sleep.

OBJECTIVE/BACKGROUND: Self-reported sleep difficulties, such as insomnia symptoms, have been reported among adolescents. Yet, studies of their prevalence and correlates are scarce among Latin Americans. This study sought (1) to describe associations between sociodemographic and lifestyle factors with self-reported sleep difficulties and (2) to examine associations between self-reported sleep difficulties and actigraphy-based sleep. PARTICIPANTS: Participants included 477 Mexican adolescents from the ELEMENT cohort. METHODS: Over 7 days, self-reported sleep measures (hard time falling asleep, overall sleep difficulties, and specific types of sleep difficulties) were obtained from daily sleep diaries. Actigraphy-based sleep measures (duration, i.e. sleep onset to morning wake, midpoint, and fragmentation) were concurrently assessed using a wrist actigraph. RESULTS: Mean (SD) age was 15.9 (2.2) years, and 53.5% were females. Mean (SD) sleep duration was 8.5 (1.2) h/night. Half reported a hard time falling asleep at least 3 days, and 25% had sleep difficulties at least 3 days over 7 days. The 3 types of sleep difficulties commonly reported among the entire cohort were insomnia/restlessness (29%), environmental (27%), and mental/emotional difficulties (19%). Female sex, smoking behavior, and socioeconomic indicators were among the most consistent factors associated with sleep difficulties. Subjective sleep difficulties were associated with shorter sleep duration (ß = -20.8 [-35.3, -6.2] min), while subjective hard time falling asleep was associated with longer sleep duration (ß = 11.3 [4.6, 27.2] min). CONCLUSION: A high proportion of Mexican adolescents in the sample reported sleep difficulties. Findings demonstrate the importance of obtaining subjective and objective sleep measures for a more comprehensive assessment of adolescent sleep.

Amyloidogenic Processing of Amyloid Precursor Protein Drives Stretch-Induced Disruption of Axonal Transport in hiPSC-Derived Neurons.

Traumatic brain injury (TBI) results in disrupted brain function following impact from an external force and is a risk factor for sporadic Alzheimer's disease (AD). Although neurologic symptoms triggered by mild traumatic brain injuries (mTBI), the most common form of TBI, typically resolve rapidly, even an isolated mTBI event can increase the risk to develop AD. Aberrant accumulation of amyloid ß peptide (Aß), a cleaved fragment of amyloid precursor protein (APP), is a key pathologic outcome designating the progression of AD following mTBI and has also been linked to impaired axonal transport. However, relationships among mTBI, amyloidogenesis, and axonal transport remain unclear, in part because of the dearth of human models to study the neuronal response following mTBI. Here, we implemented a custom-microfabricated device to deform neurons derived from human-induced pluripotent stem cells, derived from a cognitively unimpaired male individual, to mimic the mild stretch experienced by neurons during mTBI. Although no cell lethality or cytoskeletal disruptions were observed, mild stretch was sufficient to stimulate rapid amyloidogenic processing of APP. This processing led to abrupt cessation of APP axonal transport and progressive formation of aberrant axonal accumulations that contained APP, its processing machinery, and amyloidogenic fragments. Consistent with this sequence of events, stretch-induced defects were abrogated by reducing amyloidogenesis either pharmacologically or genetically. In sum, we have uncovered a novel and manipulable stretch-induced amyloidogenic pathway directly responsible for APP axonal transport dysregulation. Our findings may help to understand and ultimately mitigate the risk of developing AD following mTBI.SIGNIFICANCE STATEMENT Mild traumatic brain injury is a risk factor for sporadic Alzheimer's disease (AD). Increased amyloid ß peptide generation after injury may drive this risk. Here, by using a custom-built device to impose mild stretch to human neurons, we found that stretch triggers amyloid precursor protein (APP) cleavage, and thus amyloid ß peptide generation, consequently disrupting APP axonal transport. Compellingly, protecting APP from cleavage was sufficient to spare axonal transport dysregulation and the consequent aberrant axonal accumulation of APP. Supporting such protective mechanism, the expression of the AD-protective APPA673T genetic variant conferred protection against stretch-induced APP axonal transport phenotypes. Our data reveal potential subcellular pathways contributing to the development of AD-associated phenotypes following mild traumatic brain injury, and putative strategies for intervening in these pathways.

Desafíos de medición para la investigación de la obesidad infantil en y entre América Latina y Estados Unidos.

La obesidad infantil es un grave problema de salud pública en Latinoamérica y Estados Unidos y para luchar contra ella se precisan instrumentos de medición válidos y fiables y que tengan en cuenta los aspectos culturales. Para avanzar en este campo sería útil mejorar las mediciones en los distintos grupos de edades y países del continente americano, tanto en las comunidades de origen como en las de acogida. Además, una mayor precisión y comparabilidad de las mediciones contribuiría a acelerar la colaboración y el aprendizaje transfronterizos. En este documento presentamos (1) los marcos conceptuales que han influido en nuestra perspectiva sobre la obesidad infantil y las necesidades relacionadas con su medición en el continente americano; (2) un resumen de los recursos y guías existentes sobre los instrumentos de medida y su adaptación a la investigación de la obesidad infantil; (3) tres áreas clave que plantean desafíos y oportunidades para avanzar en la medición de la obesidad infantil, incluidas las conductas parentales, la aculturación y el potencial de utilizar métodos etnográficos para identificar los factores clave relacionados con la economía y la globalización. El progreso para reducir la obesidad infantil en las Américas podría acelerarse mediante mayor colaboración transnacional destinada a optimizar la medición para una mejor vigilancia, desarrollo y evaluación de intervenciones, investigación de implementación y evaluación de los experimentos naturales. Además, es necesario mejorar la formación sobre los instrumentos de medición y el acceso a medidas válidas y fiables tanto en español como en otros idiomas de uso frecuente en las Américas.

Foliar Application of Copper Oxide Nanoparticles Suppresses Fusarium Wilt Development on Chrysanthemum.

Micronutrients applied as nanoparticles of metal oxides have shown efficacy in vegetable and other crops for improving yield and reducing Fusarium diseases, but their role in ornamental crop management has not been investigated. In 2017, 2018, and 2020, nanoparticles of CuO, Mn2O3, or ZnO were foliarly applied at 500 µg/mL (0.6 mg/plant) to chrysanthemum transplants and planted in potting soil noninfested or infested with Fusarium oxysporum f. sp. chrysanthemi. An untreated control and a commercial fungicide, Fludioxonil, was also included. Chrysanthemums treated with nanoscale CuO had a 55, 30, and 32% reduction in disease severity ratings compared to untreated plants in 2017, 2018, and 2020, respectively. Specifically, the average dry biomass for the three years was reduced 22% by disease, but treatment with nanoscale CuO led to a 23% increase when compared to controls. Similar trends with plant height were observed. Horticultural quality was improved 28% with nano CuO and was equal to the fungicide. Nanoscale Mn2O3 and the fungicide did not consistently reduce disease ratings or increase dry biomass each year. Nanoscale ZnO was ineffective. Nanoscale CuO-treated plants had 24 to 48% more Cu/g tissue than controls (P < 0.001). These findings agree with past reports on food crops where single applications of nanoscale CuO improved plant health, growth, and yield and could offer significant impacts for managing plant diseases on ornamentals.

Measurement challenges for childhood obesity research within and between Latin America and the United States.

Childhood obesity is a major public health challenge across Latin America and the United States. Addressing childhood obesity depends on valid, reliable, and culturally sensitive measurements. Such progress within and between countries of the Americas could be enhanced through better measurement across different age groups, different countries, and in sending and receiving communities. Additionally, better and more comparable measurements could accelerate cross-border collaboration and learning. Here, we present (1) frameworks that influenced our perspectives on childhood obesity and measurement needs across the Americas; (2) a summary of resources and guidance available concerning measurement and adaptation of measures for childhood obesity research; and (3) three major areas that present challenges and opportunities for measurement advances related to childhood obesity, including parental behavior, acculturation, and the potential to incorporate ethnographic methods to identify critical factors related to economics and globalization. Progress to reduce childhood obesity across the Americas could be accelerated by further transnational collaboration aimed at improving measurement for better surveillance, intervention development and evaluation, implementation research, and evaluation of natural experiments. Additionally, there is a need to improve training related to measurement and for improving access to valid and reliable measures in Spanish and other languages common in the Americas.

Diverse Molecular Mechanisms Contribute to Differential Expression of Human Duplicated Genes.

Emerging evidence links genes within human-specific segmental duplications (HSDs) to traits and diseases unique to our species. Strikingly, despite being nearly identical by sequence (>98.5%), paralogous HSD genes are differentially expressed across human cell and tissue types, though the underlying mechanisms have not been examined. We compared cross-tissue mRNA levels of 75 HSD genes from 30 families between humans and chimpanzees and found expression patterns consistent with relaxed selection on or neofunctionalization of derived paralogs. In general, ancestral paralogs exhibited greatest expression conservation with chimpanzee orthologs, though exceptions suggest certain derived paralogs may retain or supplant ancestral functions. Concordantly, analysis of long-read isoform sequencing data sets from diverse human tissues and cell lines found that about half of derived paralogs exhibited globally lower expression. To understand mechanisms underlying these differences, we leveraged data from human lymphoblastoid cell lines (LCLs) and found no relationship between paralogous expression divergence and post-transcriptional regulation, sequence divergence, or copy-number variation. Considering cis-regulation, we reanalyzed ENCODE data and recovered hundreds of previously unidentified candidate CREs in HSDs. We also generated large-insert ChIP-sequencing data for active chromatin features in an LCL to better distinguish paralogous regions. Some duplicated CREs were sufficient to drive differential reporter activity, suggesting they may contribute to divergent cis-regulation of paralogous genes. This work provides evidence that cis-regulatory divergence contributes to novel expression patterns of recent gene duplicates in humans.

Human Ex vivo Wound Model and Whole-Mount Staining Approach to Accurately Evaluate Skin Repair.

Chronic non-healing wounds, which primarily affect the elderly and diabetic, are a significant area of clinical unmet need. Unfortunately, current chronic wound treatments are inadequate, while available pre-clinical models poorly predict the clinical efficacy of new therapies. Here we describe a high throughput, pre-clinical model to assess multiple aspects of the human skin repair response. Partial thickness wounds were created in human ex vivo skin and cultured across a healing time course. Skin wound biopsies were collected in fixative for the whole-mount staining procedure. Fixed samples were blocked and incubated in primary antibody, with detection achieved via fluorescently conjugated secondary antibody. Wounds were counterstained and imaged via confocal microscopy before calculating percentage wound closure (re-epithelialization) in each biopsy. Applying this protocol, we reveal that 2 mm excisional wounds created in healthy donor skin are fully re-epithelialized by day 4-5 post-wounding. On the contrary, closure rates of diabetic skin wounds are significantly reduced, accompanied by perturbed barrier reformation. Combining human skin wounding with a novel whole-mount staining approach allows a rapid and reproducible method to quantify ex vivo wound repair. Collectively, this protocol provides a valuable human platform to evaluate the effectiveness of potential wound therapies, transforming pre-clinical testing and validation.

Cellular benefits of single-use negative pressure wound therapy demonstrated in a novel ex vivo human skin wound model.

Negative pressure wound therapy is a widely used treatment for chronic, nonhealing wounds. Surprisingly, few studies have systematically evaluated the cellular and molecular effects of negative pressure treatment on human skin. In addition, no study to date has directly compared recently available single-use negative pressure modalities to traditional negative pressure devices in a controlled setting. Here we developed a novel large-scale ex vivo human skin culture system to effectively evaluate the efficacy of two different negative pressure wound therapy modalities. Single-use and traditional negative pressure devices were applied to human ex vivo wounded skin sheets cultured over a period of 48 hours. Cellular tissue response to therapy was evaluated via a combination of histological analysis and transcriptional profiling, in samples collected from the wound edge, skin adjacent to the wound, and an extended skin region. Single-use negative pressure wound therapy caused less damage to wound edge tissue than traditional application, demonstrated by improved skin barrier, reduced dermal-epidermal junction disruption and a dampened damage response. Transcriptional profiling confirmed significantly less activation of multiple pro-inflammatory markers in wound edge skin treated with single-use vs traditional negative pressure therapy. These findings may help to explain the greater efficacy of sNPWT in the clinic, while offering a noninvasive system to develop improved NPWT-based therapies.

Perturbation of in vivo Neural Activity Following α-Synuclein Seeding in the Olfactory Bulb.

BACKGROUND: Parkinson's disease (PD) neuropathology is characterized by intraneuronal protein aggregates composed of misfolded α-Synuclein (α-Syn), as well as degeneration of substantia nigra dopamine neurons. Deficits in olfactory perception and aggregation of α-Syn in the olfactory bulb (OB) are observed during early stages of PD, and have been associated with the PD prodrome, before onset of the classic motor deficits. α-Syn fibrils injected into the OB of mice cause progressive propagation of α-Syn pathology throughout the olfactory system and are coupled to olfactory perceptual deficits. OBJECTIVE: We hypothesized that accumulation of pathogenic α-Syn in the OB impairs neural activity in the olfactory system. METHODS: To address this, we monitored spontaneous and odor-evoked local field potential dynamics in awake wild type mice simultaneously in the OB and piriform cortex (PCX) one, two, and three months following injection of pathogenic preformed α-Syn fibrils in the OB. RESULTS: We detected α-Syn pathology in both the OB and PCX. We also observed that α-Syn fibril injections influenced odor-evoked activity in the OB. In particular, α-Syn fibril-injected mice displayed aberrantly high odor-evoked power in the beta spectral range. A similar change in activity was not detected in the PCX, despite high levels of α-Syn pathology. CONCLUSION: Together, this work provides evidence that synucleinopathy impacts in vivo neural activity in the olfactory system at the network-level.