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Objective: To describe the work experience and respiratory health of women coal miners in the United States using Coal Workers' Health Surveillance Program (CWHSP) data. Methods: Analysis included CWHSP participants with self-reported sex of female between January 1, 1970, and December 31, 2022, and examined radiographic surveillance, demographics, and job history. National Institute for Occupational Safety and Health-certified physicians classified chest radiographs. Results: Among 8,182 women participants, most worked <10 years and a majority reported working in non-dusty jobs. Among 3,392 with ≥1 year of coal mining tenure, 18 (0.5%) had evidence of pneumoconiosis, with no cases of progressive massive fibrosis. Conclusion: Women coal miners participating in the CWSHP had short mining careers and low pneumoconiosis prevalence. Few worked in the most dusty jobs, indicating limited exposure to coal mine dust. This underscores the need to explore women's roles in mining, and for improved gender-specific employment reporting. Such changes can enhance health and work conditions for women in male-dominated industries.
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Plant microbiomes are known to serve several important functions for their host, and it is therefore important to understand their composition as well as the factors that may influence these microbial communities. The microbiome of Thalassia testudinum has only recently been explored, and studies to-date have primarily focused on characterizing the microbiome of plants in a single region. Here, we present the first characterization of the composition of the microbial communities of T. testudinum across a wide geographical range spanning three distinct regions with varying physicochemical conditions. We collected samples of leaves, roots, sediment, and water from six sites throughout the Atlantic Ocean, Caribbean Sea, and the Gulf of Mexico. We then analyzed these samples using 16S rRNA amplicon sequencing. We found that site and region can influence the microbial communities of T. testudinum, while maintaining a plant-associated core microbiome. A comprehensive comparison of available microbial community data from T. testudinum studies determined a core microbiome composed of 14 ASVs that consisted mostly of the family Rhodobacteraceae. The most abundant genera in the microbial communities included organisms with possible plant-beneficial functions, like plant-growth promoting taxa, disease suppressing taxa, and nitrogen fixers.
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Malaria-causing Plasmodium parasites first replicate as liver stages (LS), which then seed symptomatic blood stage (BS) infection. Emerging evidence suggests that these stages impact each other via perturbation of host responses, and this influences the outcome of natural infection. We sought to understand whether the parasite stage interplay would affect live-attenuated whole parasite vaccination, since the efficacy of whole parasite vaccines strongly correlates with their extend of development in the liver. We thus investigated the impact of BS infection on LS development of genetically attenuated and wildtype parasites in female rodent malaria models and observed that for both, LS infection suffered severe suppression during concurrent BS infection. Strikingly and in contrast to previously published studies, we find that the BS-induced iron-regulating hormone hepcidin is not mediating suppression of LS development. Instead, we demonstrate that BS-induced host interferons are the main mediators of LS developmental suppression. The type of interferon involved depended on the BS-causing parasite species. Our study provides important mechanistic insights into the BS-mediated suppression of LS development. This has direct implications for understanding the outcomes of live-attenuated Plasmodium parasite vaccination in malaria-endemic areas and might impact the epidemiology of natural malaria infection.
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Hepatopatías , Vacunas contra la Malaria , Malaria , Plasmodium , Femenino , Humanos , Hepcidinas , Malaria/parasitología , HígadoRESUMEN
Climate change is altering the functioning of foundational ecosystems. While the direct effects of warming are expected to influence individual species, the indirect effects of warming on species interactions remain poorly understood. In marine systems, as tropical herbivores undergo poleward range expansion, they may change food web structure and alter the functioning of key habitats. While this process ('tropicalization') has been documented within declining kelp forests, we have a limited understanding of how this process might unfold across other systems. Here we use a network of sites spanning 23° of latitude to explore the effects of increased herbivory (simulated via leaf clipping) on the structure of a foundational marine plant (turtlegrass). By working across its geographic range, we also show how gradients in light, temperature and nutrients modified plant responses. We found that turtlegrass near its northern boundary was increasingly affected (reduced productivity) by herbivory and that this response was driven by latitudinal gradients in light (low insolation at high latitudes). By contrast, low-latitude meadows tolerated herbivory due to high insolation which enhanced plant carbohydrates. We show that as herbivores undergo range expansion, turtlegrass meadows at their northern limit display reduced resilience and may be under threat of ecological collapse.
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Ecosistema , Herbivoria , Cadena Alimentaria , Bosques , Cambio Climático , PlantasRESUMEN
OBJECTIVE: The aim of the study is to summarize Coal Workers' Health Surveillance Program findings since 2014, focusing on prevalence of radiographic pneumoconiosis and abnormal spirometry by region. METHODS: Analysis included the most recent Coal Workers' Health Surveillance Program encounter for working miners during October 1, 2014-June 30, 2022. Central Appalachia consists of Kentucky, Virginia, and West Virginia. RESULTS: Pneumoconiosis prevalence was highest among underground miners, with 318 (6.2%) having radiographic evidence of disease. Central Appalachian miners were more likely to have evidence of pneumoconiosis (relative risk = 4.1 [3.4-5.0]) and abnormal spirometry (relative risk = 1.4 [1.2-1.6]) compared with all others. CONCLUSIONS: Central Appalachia is a hotspot for pneumoconiosis, as well as for other indicators of respiratory impairment in coal miners. Outreach for occupational respiratory health surveillance should focus on those areas most heavily affected, ensuring that miners are not hindered by perceived or actual barriers to this secondary intervention.
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Minas de Carbón , Neumoconiosis , Humanos , Evaluación de Síntomas , Neumoconiosis/diagnóstico por imagen , Radiografía , Espirometría , Prevalencia , Carbón MineralRESUMEN
BACKGROUND: Infection precautions (IP) facilitate standardized and safe patient care. Research has demonstrated several barriers to IP adherence among health care personnel (HCP) but potential exposure risk to SARS-CoV-2 and job role has not been considered. METHODS: Researchers used self-reported baseline surveys with 191 HCPs at a university medical center to examine factors that may have affected IP adherence (eg, personal protective equipment [PPE] and hand hygiene errors) over the 2 weeks prior to the survey. Chi-square tests were used to determine if differences existed first, among job role and IP adherence, and second, the potential risk of exposure to SARS-CoV-2 and IP adherence. A binary logistic regression estimated if PPE nonadherence was associated with COVID-19 stress, job role, and potential exposure risk to SARS-CoV-2. RESULTS: PPE nonadherence varied by job role. Those in the Other group (ie, nonphysician/non-nursing HCP) reported significantly fewer errors (9.6%) compared to Physicians (26.5%) and Registered Nurses (33.3%). Hand/glove hygiene errors between COVID-19 patient rooms varied by job role. Respondents who had higher risks of exposure to SARS-CoV-2 were 5.74 times more likely to experience errors. CONCLUSIONS: The results provide implications for adopting systems-level approaches to support worker knowledge and engagement across job roles to improve IP adherence.
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COVID-19 , SARS-CoV-2 , Humanos , COVID-19/epidemiología , COVID-19/prevención & control , Transmisión de Enfermedad Infecciosa de Paciente a Profesional/prevención & control , Equipo de Protección Personal , Personal de SaludRESUMEN
A working group from the Global Library of Underwater Biological Sounds effort collaborated with the World Register of Marine Species (WoRMS) to create an inventory of species confirmed or expected to produce sound underwater. We used several existing inventories and additional literature searches to compile a dataset categorizing scientific knowledge of sonifery for 33,462 species and subspecies across marine mammals, other tetrapods, fishes, and invertebrates. We found 729 species documented as producing active and/or passive sounds under natural conditions, with another 21,911 species deemed likely to produce sounds based on evaluated taxonomic relationships. The dataset is available on both figshare and WoRMS where it can be regularly updated as new information becomes available. The data can also be integrated with other databases (e.g., SeaLifeBase, Global Biodiversity Information Facility) to advance future research on the distribution, evolution, ecology, management, and conservation of underwater soniferous species worldwide.
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Biodiversidad , Ecología , Animales , Cetáceos , Peces , SonidoRESUMEN
Despite the importance of acoustic signaling in fishes, the prevalence of the behavioral contexts associated with their active (i.e., intentional) sound production remains unclear. A systematized review was conducted to explore documented acoustic behaviors in marine, subtropical fishes and potential influences affecting their relative pervasiveness. Data were collected on 186 actively soniferous fish species studied across 194 publications, identified based on existing FishSounds and FishBase datasets. Disturbance was the most common behavioral context associated with active sound production-reported for 140 species or 75% of the species studied-and then aggression (n = 46 species, 25%) and reproduction (n = 34 species, 18%). This trend, however, somewhat differed when examined by research effort, study environment, and fish family, such as reproductive sounds being more commonly reported by studies conducted in the wild. The synthesis of fish sound production behaviors was in some ways stymied by the fact that many species' sound production did not have discernible associated behavioral contexts and that some investigations did not clearly identify the study environments in which active sound production was observed. These findings emphasize the importance of context-behavioral or otherwise-when studying acoustic behaviors in fishes.
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Acústica , Sonido , Animales , Peces , AgresiónRESUMEN
Pelvic organ prolapse (POP) and stress urinary incontinence (SUI) are two common disorders that affect the anterior compartment of the pelvic floor in women. These can be treated conservatively or surgically. Among patients treated surgically, a substantial number present with pain, recurrent POP or SUI, or other conditions that warrant additional interventions. In many of these cases, imaging is key to identifying and characterizing the type of procedure performed, locating synthetic materials that may have been placed, and characterizing complications. Imaging may be particularly helpful when prior surgical records are not available or a comprehensive physical examination is not possible. US and MRI are the most commonly used modalities for such patients, although radiopaque surgical materials may be visible at voiding cystourethrography and CT. The authors summarize commonly used surgical treatment options for patients with SUI and POP, review imaging techniques for evaluation of such patients, and describe the normal imaging appearance and complications of pelvic floor surgical repair procedures in the anterior compartment of the pelvis. ©RSNA, 2023 Quiz questions for this article are available in the supplemental material.
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Prolapso de Órgano Pélvico , Procedimientos de Cirugía Plástica , Incontinencia Urinaria de Esfuerzo , Humanos , Femenino , Diafragma Pélvico/diagnóstico por imagen , Diafragma Pélvico/cirugía , Prolapso de Órgano Pélvico/diagnóstico por imagen , Prolapso de Órgano Pélvico/cirugía , Incontinencia Urinaria de Esfuerzo/diagnóstico por imagen , Incontinencia Urinaria de Esfuerzo/cirugía , Incontinencia Urinaria de Esfuerzo/complicaciones , Imagen MultimodalRESUMEN
BACKGROUND: Mandatory examination requirements for US coal miners newly entering the workforce have been in place since the 1969 Coal Act mandated chest radiographs and were updated to include spirometry with promulgation of the 2014 Mine Safety and Health Administration Dust Rule. Compliance with the mandatory respiratory screening series is described using data from the National Institute for Occupational Safety and Health Coal Workers' Health Surveillance Program (CWHSP). METHODS: Among all radiographic and spirometry submissions to the CWHSP during 30 June 1971-15 March 2022, new underground coal miners who began work in the industry after 30 June 1971, and new underground, surface miners and contractors who began work after new regulations were implemented 1 August 2014, were identified and included in analysis. RESULTS: Of the 115 093 unique miners who participated in the CWHSP and whose estimated entry into mining occurred during 30 June 1971-15 March 2019, 50 487 (43.9%) received their initial mandatory radiograph, and 15 452 (13.4%) submitted their initial and 3-year mandatory radiographs. Since new regulations were implemented, compliance with initial radiographs appeared to improve (80%) but compliance with 3-year radiographs remained low (11.6%). Compliance with spirometry testing was also low for initial (17.1%) and follow-up screenings (2.7%). CONCLUSIONS: The majority of new coal miners eligible for health surveillance did not receive a baseline radiograph or spirometry test through the CWHSP even though coal mine operators are required by law to provide these. Ensuring coal miners' regular participation in health surveillance from early in their careers is an important way to monitor and protect their respiratory health.
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Minas de Carbón , Mineros , Neumoconiosis , Estados Unidos , Humanos , Polvo , Tamizaje Masivo , Espirometría , Carbón Mineral , Neumoconiosis/epidemiologíaRESUMEN
Malaria-causing Plasmodium parasites have a complex life cycle and present numerous antigen targets that may contribute to protective immune responses. The currently recommended vaccine-RTS,S-functions by targeting the Plasmodium falciparum circumsporozoite protein (CSP), which is the most abundant surface protein of the sporozoite form responsible for initiating infection of the human host. Despite showing only moderate efficacy, RTS,S has established a strong foundation for the development of next-generation subunit vaccines. Our previous work characterizing the sporozoite surface proteome identified additional non-CSP antigens that may be useful as immunogens individually or in combination with CSP. In this study, we examined eight such antigens using the rodent malaria parasite Plasmodium yoelii as a model system. We demonstrate that despite conferring weak protection individually, coimmunizing each of several of these antigens alongside CSP could significantly enhance the sterile protection achieved by CSP immunization alone. Thus, our work provides compelling evidence that a multiantigen preerythrocytic vaccine approach may enhance protection compared to CSP-only vaccines. This lays the groundwork for further studies aimed at testing the identified antigen combinations in human vaccination trials that assess efficacy with controlled human malaria infection. IMPORTANCE The currently approved malaria vaccine targets a single parasite protein (CSP) and results in only partial protection. We tested several additional vaccine targets in combination with CSP to identify those that could enhance protection from infection upon challenge in the mouse malaria model. In identifying several such enhancing vaccine targets, our work indicates that a multiprotein immunization approach may be a promising avenue to achieving higher levels of protection from infection. Our work identified several candidate leads for follow-up in the models relevant for human malaria and provides an experimental framework for efficiently carrying out such screens for other combinations of vaccine targets.
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INTRODUCTION: The Hall technique is a biological caries management approach commonly used within the field of paediatric dentistry. The technique involves the placement of a preformed metal crown (PMC) without local anaesthetic, caries removal or tooth preparation. Despite being widely taught across dental schools in the United Kingdom, evidence from the literature suggests that PMCs are seldom used in general dental practice, even by newly qualified dentists. This study aimed to evaluate the experience and self-reported confidence of Foundation Dentists (FDs) in the North West of England in the placement of preformed metal crowns using the Hall technique. MATERIALS AND METHODS: An anonymous questionnaire was distributed to 117 Foundation Dentists in the North West of England. RESULTS: Forty-six questionnaires were returned, giving a response rate of 39%. The results revealed that 80.4% of FDs felt "somewhat," "quite" or "extremely" confident with the Hall technique at the point of graduation. This increased to 85.3% upon completion of Dental Foundation Training. CONCLUSION: Foundation Dentists' self-reported confidence in the Hall technique was reasonably high both at the point of qualification and upon completing Dental Foundation Training. Nevertheless, many of the Foundation Dentists felt that they would benefit from further postgraduate training on the technique.
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Caries Dental , Diente Primario , Niño , Humanos , Educación en Odontología , Inglaterra , Caries Dental/terapia , Encuestas y Cuestionarios , Coronas , Metales , Odontólogos , Pautas de la Práctica en OdontologíaRESUMEN
Background: Retinoblastoma is a rare form of pediatric eye cancer for which enucleation is a common treatment modality. There is an increasing focus upon the impact of enucleation on children and families. This study aimed to explore the experiences of children and their families following enucleation to consider the barriers that may be encountered when adjusting to living with an artificial eye and identifying the support services and strategies used to address these barriers. Methods: Using a descriptive qualitative approach, interviews were conducted with 12 parents and seven children and thematic analysis was used to identify four themes representing the perspective of parents and children. Results: Parent themes identified were (a) entry into the world of retinoblastoma; (b) the importance of specialist support; (c) a family learning to cope; and (d) navigating school. The perspectives of children were (a) the importance of preparation and play; (b) positive reinforcement and hospital support; (c) support and openness at home; and (d) the importance of good school planning. Two overarching themes related to parent and child adaptation were also identified. Discussion: The study findings reveal that the process of adapting to living with an artificial eye extended to influencing decisions around parenting, learning to advocate for their child, and supporting children through school and peer relationships. For children, the importance of ongoing support and information was vital to enhance understanding, adaptation, and development of independence. Overall, the study suggests that children and families undergoing enucleation need continuing, individualized, and specialist support.
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Neoplasias de la Retina , Retinoblastoma , Niño , Ojo Artificial , Humanos , Responsabilidad Parental , Padres , Investigación Cualitativa , Neoplasias de la Retina/cirugía , Retinoblastoma/cirugíaRESUMEN
Neuropathic pain affects millions of people worldwide, yet the molecular mechanisms of how it develops and persists are poorly understood. Given that males have historically been utilized as the primary sex in preclinical studies, less is known about the female neuroinflammatory response to injury, formation of pain, or response to pain-relieving therapies. Macrophages contribute to the development of neuroinflammatory pain via the activation of their cyclooxygenase-2 (COX-2) enzyme, which leads to the production of prostaglandin E2 (PGE2). PGE2 activates nociception and influences additional leukocyte infiltration. Attenuation of COX-2 activity decreases inflammatory pain, most commonly achieved by nonsteroidal anti-inflammatory drugs (NSAIDs), yet NSAIDs are considered ineffective for neuropathic pain due to off target toxicity. Using chronic constriction injury of the rat sciatic nerve, we show that males and females exhibit quantitatively the same degree of mechanical allodynia post injury. Furthermore, a low-dose nanotherapeutic containing the NSAID celecoxib is phagocytosed by circulating monocytes that then naturally accumulate at sites of injury as macrophages. Using this nanotherapeutic, we show that treated males exhibit complete reversal of hypersensitivity, while the same dose of nanotherapeutic in females provides an attenuated relief. The difference in behavioral response to the nanotherapy is reflected in the reduction of infiltrating macrophages at the site of injury. The observations contained in this study reinforce the notion that female neuroinflammation is different than males.
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Neuralgia , Caracteres Sexuales , Animales , Antiinflamatorios no Esteroideos/farmacología , Antiinflamatorios no Esteroideos/uso terapéutico , Celecoxib/farmacología , Celecoxib/uso terapéutico , Ciclooxigenasa 2 , Femenino , Humanos , Masculino , Neuralgia/tratamiento farmacológico , Enfermedades Neuroinflamatorias , Prostaglandinas E , RatasRESUMEN
BACKGROUND: Dihydrocodeine (DHC) is considered a 'weak' opioid, but there is evidence of its increasing misuse in overdose deaths. This research aims to analyse trends in DHC-related deaths in England relevant to source and dose of DHC, and decedent demographics. METHODS: Cases from England reported to the National Programme on Substance Abuse Deaths (NPSAD) where DHC was identified at post-mortem and/or implicated in death between 2001 and 2020 were extracted for analysis. RESULTS: 2071 DHC-related deaths were identified. The greatest number of deaths involved illicitly obtained DHC and a significant increase in these deaths was recorded over time (r = 0.5, p = 0.03). However, there was a concurrent decline in the implication rate of DHC in causing death (r = -0.6, p < 0.01). Fatalities were primarily due to accidental overdose (64.8%) and misuse was highly prevalent in combination with additional central nervous system depressants (95.3%), namely illicit heroin/morphine and diazepam. In contrast, when DHC was obtained over-the-counter (OTC) suicide mortality accounted for almost half of the deaths (42.5%). Differences in polysubstance use were also identified, with less heroin/morphine and benzodiazepine co-detection, but increased OTC codeine co-detection. CONCLUSIONS: DHC misuse in England is increasing. The pharmacological consideration of DHC as a 'weak' opioid may be misinterpreted by users, leading to accidental overdosing. There is an urgent need to understand increasing polypharmacy in overdose deaths. Additionally, suicides involving DHC is a potential cause for concern and a review of OTC opioid-paracetamol preparations is necessary to determine whether the benefits of these medications continue to outweigh the risks of intentional overdose.
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Sobredosis de Droga , Trastornos Relacionados con Sustancias , Suicidio , Analgésicos Opioides/efectos adversos , Codeína/efectos adversos , Codeína/análogos & derivados , Codeína/análisis , Inglaterra/epidemiología , Heroína , Humanos , Morfina , Trastornos Relacionados con Sustancias/tratamiento farmacológicoRESUMEN
OBJECTIVE: To identify potential strategies to mitigate COVID-19 transmission in a Utah meat-processing facility and surrounding community. DESIGN/SETTING: During March-June 2020, 502 workers at a Utah meat-processing facility (facility A) tested positive for SARS-CoV-2. Using merged data from the state disease surveillance system and facility A, we analyzed the relationship between SARS-CoV-2 positivity and worker demographics, work section, and geospatial data on worker residence. We analyzed worker survey responses to questions regarding COVID-19 knowledge, beliefs, and behaviors at work and home. PARTICIPANTS: (1) Facility A workers (n = 1373) with specimen collection dates and SARS-CoV-2 RT-PCR test results; (2) residential addresses of all persons (workers and nonworkers) with a SARS-CoV-2 diagnostic test (n = 1036), living within the 3 counties included in the health department catchment area; and (3) facility A workers (n = 64) who agreed to participate in the knowledge, attitudes, and practices survey. MAIN OUTCOME MEASURES: New cases over time, COVID-19 attack rates, worker characteristics by SARS-CoV-2 test results, geospatially clustered cases, space-time proximity of cases among workers and nonworkers; frequency of quantitative responses, crude prevalence ratios, and counts and frequency of coded responses to open-ended questions from the COVID-19 knowledge, attitudes, and practices survey. RESULTS: Statistically significant differences in race (P = .01), linguistic group (P < .001), and work section (P < .001) were found between workers with positive and negative SARS-CoV-2 test results. Geographically, only 6% of cases were within statistically significant spatiotemporal case clusters. Workers reported using handwashing (57%) and social distancing (21%) as mitigation strategies outside work but reported apprehension with taking COVID-19-associated sick leave. CONCLUSIONS: Mitigating COVID-19 outbreaks among workers in congregate settings requires a multifaceted public health response that is tailored to the workforce. IMPLICATIONS FOR POLICY AND PRACTICE: Tailored, multifaceted mitigation strategies are crucial for reducing COVID-19-associated health disparities among disproportionately affected populations.
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COVID-19 , Brotes de Enfermedades , Humanos , Carne , Salud Pública , SARS-CoV-2 , Utah/epidemiologíaRESUMEN
OBJECTIVES: Rates of nonalcoholic fatty liver disease (NAFLD) are increasing worldwide. The fatty liver index (FLI) is a noninvasive predictor of NAFLD. This prospective cohort study used the FLI to estimate the prevalence of NAFLD in patients attending an Irish Acute Medical Unit (AMU), and assessed the degree of fibrosis in this group using Fibroscan. METHODS: Patients attending the AMU over a 3-month period were invited to participate. Patients with excess alcohol consumption or pre-existing liver disease were excluded. Using established FLI cut-offs, 414 participants were grouped into low (FLI ≤ 30), medium (30 < FLI ≤ 60) and high (FLI > 60) risk of NAFLD. High-risk patients were offered review including liver stiffness measurement (LSM) and controlled attenuation parameter (CAP) score. RESULTS: A total of 134 patients were at low-risk, 96 at medium-risk and 184 at high-risk of NAFLD. Male sex (P < 0.0001) and increasing age (P < 0.0001) were associated with higher risk. Of the 120 high-risk patients who attended follow up, 13 participants had LSM > 7 kPa. Higher FLI scores were associated with higher CAP scores (P < 0.0001) but did not predict higher LSMs. Fasting glucose and HbA1c were found to be associated with higher LSM. CONCLUSION: About 44.4% of patients presenting to the AMU were at high risk of NAFLD according to the FLI. Only 10.8% of the high-risk group, and 3% of all those recruited had a LSM > 7 kPa suggesting development of fibrosis.
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Diagnóstico por Imagen de Elasticidad , Enfermedad del Hígado Graso no Alcohólico , Humanos , Hígado/diagnóstico por imagen , Hígado/patología , Cirrosis Hepática/complicaciones , Cirrosis Hepática/diagnóstico por imagen , Cirrosis Hepática/epidemiología , Masculino , Enfermedad del Hígado Graso no Alcohólico/complicaciones , Enfermedad del Hígado Graso no Alcohólico/diagnóstico por imagen , Enfermedad del Hígado Graso no Alcohólico/epidemiología , Prevalencia , Estudios ProspectivosRESUMEN
Gametocytes of the malaria parasite Plasmodium are taken up by the mosquito vector with an infectious blood meal, representing a critical stage for parasite transmission. Calcium-independent protein kinases (CDPKs) play key roles in calcium-mediated signaling across the complex life cycle of the parasite. We sought to understand their role in human parasite transmission from the host to the mosquito vector and thus investigated the role of the human-infective parasite Plasmodium falciparum CDPK4 in the parasite life cycle. P. falciparum cdpk4- parasites created by targeted gene deletion showed no effect in blood stage development or gametocyte development. However, cdpk4- parasites showed a severe defect in male gametogenesis and the emergence of flagellated male gametes. To understand the molecular underpinnings of this defect, we performed mass spectrometry-based phosphoproteomic analyses of wild-type and Plasmodium falciparum cdpk4- late gametocyte stages to identify key CDPK4-mediated phosphorylation events that may be important for the regulation of male gametogenesis. We further employed in vitro assays to identify these putative substrates of Plasmodium falciparum CDPK4. This indicated that CDPK4 regulates male gametogenesis by directly or indirectly controlling key essential events, such as DNA replication, mRNA translation, and cell motility. Taken together, our work demonstrates that PfCDPK4 is a central kinase that regulates exflagellation and thereby is critical for parasite transmission to the mosquito vector. IMPORTANCE Transmission of the malaria parasite to the mosquito vector is critical for the completion of the sexual stage of the parasite life cycle and is dependent on the release of male gametes from the gametocyte body inside the mosquito midgut. In the present study, we demonstrate that PfCDPK4 is critical for male gametogenesis and is involved in phosphorylation of proteins essential for male gamete emergence. Targeting PfCDPK4 and its substrates may provide insights into achieving effective malaria transmission-blocking strategies.
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Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , Gametogénesis/fisiología , Mosquitos Vectores , Plasmodium falciparum/enzimología , Plasmodium falciparum/metabolismo , Animales , Señalización del Calcio , Proteínas Quinasas Dependientes de Calcio-Calmodulina/genética , Culicidae , Gametogénesis/genética , Células Germinativas/metabolismo , Estadios del Ciclo de Vida , Malaria Falciparum/parasitología , Malaria Falciparum/transmisión , Masculino , Fosforilación , Plasmodium falciparum/genética , Proteínas Protozoarias/genéticaRESUMEN
INTRODUCTION: Because of phenotypic overlap between monogenic urinary stone diseases (USD), gene-specific analyses can result in missed diagnoses. We used targeted next generation sequencing (tNGS), including known and candidate monogenic USD genes, to analyze suspected primary hyperoxaluria (PH) or Dent disease (DD) patients genetically unresolved (negative; N) after Sanger analysis of the known genes. Cohorts consisted of 285 PH (PHN) and 59 DD (DDN) families. METHODS: Variants were assessed using disease-specific and population databases plus variant assessment tools and categorized using the American College of Medical Genetics (ACMG) guidelines. Prior Sanger analysis identified 47 novel PH or DD gene pathogenic variants. RESULTS: Screening by tNGS revealed pathogenic variants in 14 known monogenic USD genes, accounting for 45 families (13.1%), 27 biallelic and 18 monoallelic, including 1 family with a copy number variant (CNV). Recurrent genes included the following: SLC34A3 (n = 13), CLDN16 (n = 8), CYP24A1 (n = 4), SLC34A1 (n = 3), SLC4A1 (n = 3), APRT (n = 2), CLDN19 (n = 2), HNF4A1 (n = 2), and KCNJ1 (n = 2), whereas ATP6V1B1, CASR, and SLC12A1 and missed CNVs in the PH genes AGXT and GRHPR accounted for 1 pedigree each. Of the 48 defined pathogenic variants, 27.1% were truncating and 39.6% were novel. Most patients were diagnosed before 18 years of age (76.1%), and 70.3% of biallelic patients were homozygous, mainly from consanguineous families. CONCLUSION: Overall, in patients suspected of DD or PH, 23.9% and 7.3% of cases, respectively, were caused by pathogenic variants in other genes. This study shows the value of a tNGS screening approach to increase the diagnosis of monogenic USD, which can optimize therapies and facilitate enrollment in clinical trials.