Dynamic and broad expression of adamts9 in developing and adult zebrafish.

BACKGROUND: Previous studies showed that Adamts9 is involved in multiple functions including ovulation, spine formation, primordial germ cell migration, and development of primary ovarian follicles in animals. However, systemic examination and high-resolution analyses of adamts9 expression are missing due to lack of a sensitive reporter assay. RESULTS: In the present study, we created a new transgenic zebrafish reporter line Tg(adamts9:EGFP) and assayed its expression in various tissues and cells during development and in adults at high-resolution using confocal imaging. Reporter expression was validated with real-time quantitative PCR, whole mount in situ hybridization, and immunohistochemistry for endogenous adamts9. Strong expression of the adamts9:EGFP transgene was found in a wide range of adult and embryonic zebrafish tissues/cells including ovaries, testes, brains, eyes, pectoral fins, intestine, skin, gill, muscle, and heart; while lower expression was observed in the liver and growing ovarian follicles (stages II and III). CONCLUSIONS: Our results of a broad and dynamic expression pattern for this evolutionary conserved metalloprotease suggest involvement of adamts9 in the development and physiological functions of various tissues in animals.

Pediatric Eosinophilia: A Review and Multiyear Investigation into Etiologies.

OBJECTIVES: To identify the etiology of peripheral eosinophilia in a large pediatric population and to develop a diagnostic algorithm to help guide diagnosis and management of peripheral eosinophilia in the outpatient pediatric population. STUDY DESIGN: We performed a retrospective chart review of children presenting to Texas Children's Hospital in Houston with peripheral eosinophilia between January 1, 2011 and December 31, 2019. Eosinophilia was classified as mild (absolute eosinophil count [AEC] >500 and <1500 cells/µL), moderate (AEC >1500 and <4500 cells/µL), or severe (AEC >4500 cells/µL). Demographic information and diagnostic workup data were collected. RESULTS: A total of 771 patients aged <18 years were evaluated. The most common cause of eosinophilia was allergy (n = 357; 46%), with atopy (n = 296) and drug reaction (n = 54) the most common subcauses. This was followed by unknown etiology (n = 274; 36%), infectious causes (n = 72; 9%), and eosinophilic disorders (n = 47; 6%). Many patients with an unknown cause (n = 202; 74%) had limited or no follow-up testing. CONCLUSIONS: More information on the etiology of pediatric eosinophilia and workup data could help identify the causes. This study provides important information on the evaluation of eosinophilia in the US pediatric population, including a diagnostic algorithm to guide primary care pediatricians.

The morphological and functional diversity of apical microvilli.

Sensory neurons use specialized apical processes to perceive external stimuli and monitor internal body conditions. The apical apparatus can include cilia, microvilli, or both, and is adapted for the functions of the particular cell type. Photoreceptors detect light through a large, modified cilium (outer segment), that is supported by a surrounding ring of microvilli-like calyceal processes (CPs). Although first reported 150 years ago, CPs remain poorly understood. As a basis for future study, we therefore conducted a review of existing literature about sensory cell microvilli, which can act either as the primary sensory detector or as support for a cilia-based detector. While all microvilli are finger-like cellular protrusions with an actin core, the processes vary across cell types in size, number, arrangement, dynamics, and function. We summarize the current state of knowledge about CPs and the characteristics of the microvilli found on inner ear hair cells (stereocilia) and cerebral spinal fluid-contacting neurons, with comparisons to the brush border of the intestinal and renal epithelia. The structure, stability, and dynamics of the actin core are regulated by a complement of actin-binding proteins, which includes both common components and unique features when compared across cell types. Further, microvilli are often supported by lateral links, a glycocalyx, and a defined extracellular matrix, each adapted to the function and environment of the cell. Our comparison of microvillar features will inform further research into how CPs support photoreceptor function, and also provide a general basis for investigations into the structure and functions of apical microvilli found on sensory neurons.

A sensation for inflation: initial swim bladder inflation in larval zebrafish is mediated by the mechanosensory lateral line.

Larval zebrafish achieve neutral buoyancy by swimming up to the surface and taking in air through their mouths to inflate their swim bladders. We define this behavior as 'surfacing'. Little is known about the sensory basis for this underappreciated behavior of larval fish. A strong candidate is the mechanosensory lateral line, a hair cell-based sensory system that detects hydrodynamic information from sources such as water currents, predators, prey and surface waves. However, a role for the lateral line in mediating initial inflation of the swim bladder has not been reported. To explore the connection between the lateral line and surfacing, we used a genetic mutant (lhfpl5b-/-) that renders the zebrafish lateral line insensitive to mechanical stimuli. We observed that approximately half of these lateral line mutants over-inflate their swim bladders during initial inflation and become positively buoyant. Thus, we hypothesized that larval zebrafish use their lateral line to moderate interactions with the air-water interface during surfacing to regulate swim bladder inflation. To test the hypothesis that lateral line defects are responsible for swim bladder over-inflation, we showed that exogenous air is required for the hyperinflation phenotype and transgenic rescue of hair cell function restores normal inflation. We also found that chemical ablation of anterior lateral line hair cells in wild-type larvae causes hyperinflation. Furthermore, we show that manipulation of lateral line sensory information results in abnormal inflation. Finally, we report spatial and temporal differences in the surfacing behavior between wild-type and lateral line mutant larvae. In summary, we propose a novel sensory basis for achieving neutral buoyancy where larval zebrafish use their lateral line to sense the air-water interface and regulate initial swim bladder inflation.

Incorporating COVID-19 into Acute Febrile Illness Surveillance Systems, Belize, Kenya, Ethiopia, Peru, and Liberia, 2020-2021.

Existing acute febrile illness (AFI) surveillance systems can be leveraged to identify and characterize emerging pathogens, such as SARS-CoV-2, which causes COVID-19. The US Centers for Disease Control and Prevention collaborated with ministries of health and implementing partners in Belize, Ethiopia, Kenya, Liberia, and Peru to adapt AFI surveillance systems to generate COVID-19 response information. Staff at sentinel sites collected epidemiologic data from persons meeting AFI criteria and specimens for SARS-CoV-2 testing. A total of 5,501 patients with AFI were enrolled during March 2020-October 2021; >69% underwent SARS-CoV-2 testing. Percentage positivity for SARS-CoV-2 ranged from 4% (87/2,151, Kenya) to 19% (22/115, Ethiopia). We show SARS-CoV-2 testing was successfully integrated into AFI surveillance in 5 low- to middle-income countries to detect COVID-19 within AFI care-seeking populations. AFI surveillance systems can be used to build capacity to detect and respond to both emerging and endemic infectious disease threats.

Maintaining health professional education during war: A scoping review.

PURPOSE: War negatively impacts health professional education when health care is needed most. The aims of this scoping review are to describe the scope of barriers and targeted interventions to maintaining health professional education during war and summarise the research. METHODS: We conducted a scoping review between 20 June 2018 and 2 August 2018. The search was restricted to English publications including peer-reviewed publications without date ranges involving war and health professional education (medical school, residency training and nursing school), with interventions described to maintain educational activities. Two independent reviewers completed inclusion determinations and data abstraction. Thematic coding was performed using an inductive approach allowing dominant themes to emerge. The frequency of barrier and intervention themes and illustrative quotes were extracted. Articles were divided into modern/postmodern categories to permit temporal and historical analysis of thematic differences. RESULTS: Screening identified 3271 articles, with 56 studies meeting inclusion criteria. Publication dates ranged from 1914-2018 with 17 unique wars involving 17 countries. The studies concerned medical students (61.4%), residents (28.6%) and nursing students (10%). Half involved the modern era and half the postmodern era. Thematic coding identified five categories of barriers and targeted interventions in maintaining health professions education during war: curriculum, personnel, wellness, resources, and oversight, with most involving curriculum and personnel. The distribution of themes among various health professional trainees was similar. The frequency and specifics changed temporally reflecting innovations in medical education and war, with increased focus on oversight and personnel during the modern era and greater emphasis on wellness, curriculum, and resources during the postmodern era. CONCLUSIONS: There are overarching categories of barriers and targeted interventions in maintaining health professional education during war which evolve over time. These may serve as a useful framework to strategically support future research and policy efforts.

Pesticide Exposure and Heat Exhaustion in a Migrant Agricultural Worker: A Case of Labor Trafficking.

To our knowledge, there are no published reports of heat exhaustion and pesticide exposure in a labor trafficked patient in the literature. Here we represent the case of J.C.J.L., who was labor trafficked. He presented to a local emergency department with heat exhaustion and pesticide exposure related to working conditions in a Mississippi corn field. Unfortunately, while he received medical treatment, his labor trafficking condition was missed. Emergency departments should be equipped to assess for human trafficking and connect trafficked persons with the resources they need. Emergency physicians should maintain a high index of suspicion for human trafficking among migrant workers presenting with occupation-related complaints.

Environmental health effects attributed to toxic and infectious agents following hurricanes, cyclones, flash floods and major hydrometeorological events.

Extreme hydrometeorological events such as hurricanes and cyclones are increasing in frequency and intensity due to climate change and often associated with flash floods in coastal, urbanized and industrial areas. Preparedness and response measures need to concentrate on toxicological and infectious hazards, the potential impact on environmental health, and threat to human lives. The recognition of the danger of flood water after hurricanes is critical. Effective health management needs to consider the likelihood and specific risks of toxic agents present in waters contaminated by chemical spills, bio-toxins, waste, sewage, and water-borne pathogens. Despite significant progress in the ability to rapidly detect and test water for a wide range of chemicals and pathogens, there has been a lack of implementation to adapt toxicity measurements in the context of flash and hurricane-induced flooding. The aim of this review was to highlight the need to collect and analyze data on toxicity of flood waters to understand the risks and prepare vulnerable communities and first responders. It is proposed that new and routinely used technologies be employed during disaster response to rapidly assess toxicity and infectious disease threats, and subsequently take necessary remedial actions.

Functional Analysis of the Transmembrane and Cytoplasmic Domains of Pcdh15a in Zebrafish Hair Cells.

Protocadherin 15 (PCDH15) is required for mechanotransduction in sensory hair cells as a component of the tip link. Isoforms of PCDH15 differ in their cytoplasmic domains (CD1, CD2, and CD3), but share the extracellular and transmembrane (TMD) domains, as well as an intracellular domain known as the common region (CR). In heterologous expression systems, both the TMD and CR of PCDH15 have been shown to interact with members of the mechanotransduction complex. The in vivo significance of these protein-protein interaction domains of PCDH15 in hair cells has not been determined. Here, we examined the localization and function of the two isoforms of zebrafish Pcdh15a (CD1 and CD3) in pcdh15a-null mutants by assessing Pcdh15a transgene-mediated rescue of auditory/vestibular behavior and hair cell morphology and activity. We found that either isoform alone was able to rescue the Pcdh15a-null phenotype and that the CD1- or CD3-specific regions were dispensable for hair bundle integrity and labeling of hair cells with FM4-64, which was used as a proxy for mechanotransduction. When either the CR or TMD domain was deleted, the mutated proteins localized to the stereocilial tips, but were unable to rescue FM4-64 labeling. Disrupting both domains led to a complete failure of Pcdh15a to localize to the hair bundle. Our findings demonstrate that the TMD and cytoplasmic CR domains are required for the in vivo function of Pcdh15a in zebrafish hair cells.SIGNIFICANCE STATEMENT Tip links transmit force to mechanotransduction channels at the tip of hair bundles in sensory hair cells. One component of tip links is Protocadherin 15 (PCDH15). Here, we demonstrate that, when transgenically expressed, either zebrafish Pcdh15a-cytodomain 1 (CD1) or Pcdh15a-CD3 can rescue the phenotype of a pcdh15a-null mutant. Even when lacking the specific regions for CD1 or CD3, truncated Pcdh15a that contains the so-called common region (CR) at the cytoplasmic/membrane interface still has the ability to rescue similar to full-length Pcdh15a. In contrast, Pcdh15a lacking the entire cytoplasmic domain is not functional. These results demonstrate that the CR plays a key role in the mechanotransduction complex in hair cells.

Environmental and Ecological Effects of Climate Change on Venomous Marine and Amphibious Species in the Wilderness.

INTRODUCTION: Recent analyses of data show a warming trend in global average air and sea surface ocean temperatures. The atmosphere and ocean have warmed, the amounts of snow and ice have diminished, the sea level has risen, and the concentrations of greenhouse gases have increased. This article will focus on climate change and projected effects on venomous marine and amphibious creatures with the potential impact on human health. METHODS: Retrospective analysis of environmental, ecological, and medical literature with a focus on climate change, toxinology, and future modeling specific to venomous aquatic and amphibious creatures. Species included venomous jellyfish, poisonous fish, crown-of-thorns starfish, sea snakes, and toxic frogs. RESULTS: In several projected scenarios, rising temperatures, weather extremes, and shifts in seasons will increase poisonous population numbers, particularly with certain marine creatures like jellyfish and crown-of-thorns starfish. Habitat expansions by lionfish and sea snakes are projected to occur. These phenomena, along with increases in human populations and coastal development will likely increase human-animal encounters. Other species, particularly amphibious toxic frogs, are declining rapidly due to their sensitivity to any temperature change or subtle alterations in the stability of their environment. If temperatures continue to rise to record levels over the next decades, it is predicted that the populations of these once plentiful and critically important animals to the aquatic ecosystem will decline and their geographic distributions will shrink. CONCLUSION: Review of the literature investigating the effect and forecasts of climate change on venomous marine and amphibious creatures has demonstrated that temperature extremes and changes to climatic norms will likely have a dramatic effect on these toxicological organisms. The effects of climate change on these species through temperature alteration and rising coastal waters will influence each species differently and in turn potentially affect commercial industries, travel, tourism, and human health.

Potential Environmental and Ecological Effects of Global Climate Change on Venomous Terrestrial Species in the Wilderness.

INTRODUCTION: Climate change has been scientifically documented, and its effects on wildlife have been prognosticated. We sought to predict the overall impact of climate change on venomous terrestrial species. We hypothesize that given the close relationship between terrestrial venomous species and climate, a changing global environment may result in increased species migration, geographical redistribution, and longer seasons for envenomation, which would have repercussions on human health. METHODS: A retrospective analysis of environmental, ecological, and medical literature was performed with a focus on climate change, toxinology, and future modeling specific to venomous terrestrial creatures. Species included venomous reptiles, snakes, arthropods, spiders, and Hymenoptera (ants and bees). Animals that are vectors of hemorrhagic infectious disease (eg, mosquitos, ticks) were excluded. RESULTS: Our review of the literature indicates that changes to climatic norms will have a potentially dramatic effect on terrestrial venomous creatures. Empirical evidence demonstrates that geographic distributions of many species have already shifted due to changing climatic conditions. Given that most terrestrial venomous species are ectotherms closely tied to ambient temperature, and that climate change is shifting temperature zones away from the equator, further significant distribution and population changes should be anticipated. For those species able to migrate to match the changing temperatures, new geographical locations may open. For those species with limited distribution capabilities, the rate of climate change may accelerate faster than species can adapt, causing population declines. Specifically, poisonous snakes and spiders will likely maintain their population numbers but will shift their geographic distribution to traditionally temperate zones more often inhabited by humans. Fire ants and Africanized honey bees are expected to have an expanded range distribution due to predicted warming trends. Human encounters with these types of creatures are likely to increase, resulting in potential human morbidity and mortality. CONCLUSIONS: Temperature extremes and changes to climatic norms may have a dramatic effect on venomous terrestrial species. As climate change affects the distribution, populations, and life histories of these organisms, the chance of encounters could be altered, thus affecting human health and the survivability of these creatures.

Apostle Islands National Lakeshore: A Review of Search and Rescue and Emergency Medical Services Operations, 2006-2015.

INTRODUCTION: Apostle Islands National Lakeshore (APIS) lies at the northern tip of Wisconsin and is home to a network of 21 islands along Lake Superior. The goal of this report is to investigate search and rescue (SAR) and emergency medical services (EMS) trends at APIS in an effort to improve visitor safety and resource allocation. METHODS: This study is a retrospective analysis reviewing APIS SAR reports and annual EMS summary reports from January 1, 2006, to December 31, 2015. Information related to incident type, incident date, individual demographic characteristics and activities, injury/illness type, cost, and contributing factors were recorded and analyzed in frequency tables. RESULTS: From 2006 to 2015, APIS SAR conducted 133 total missions assisting 261 individuals-200 not injured/ill, 57 injured/ill, and 4 fatalities. Median cost per SAR incident involving aircraft totaled $21,695 (range: $2,993-141,849), whereas incidents not involving aircraft had a median cost of $363 (range: $35-8,830). Nonmotorized boating was the most common activity resulting in SAR incidents. All 4 fatalities were attributed to drowning while kayaking or swimming. Cold-related injury/illness accounted for nearly half of all injuries/illnesses (45%) with the most commonly reported contributing factor being high winds. EMS responded to a total of 134 incidents. Trauma and first aid accounted for 43% and 34% of EMS workload, respectively. CONCLUSIONS: Overall, this study highlights the hazards associated with the frigid and rough conditions of Lake Superior. The reported results aim to help APIS personnel more saliently convey risks to visitors and plan appropriately in an effort to decrease the need for future rescues.

Newly Recognized Pediatric Cases of Typhus Group Rickettsiosis, Houston, Texas, USA.

An increase in typhus group rickettsiosis and an expanding geographic range occurred in Texas, USA, over a decade. Because this illness commonly affects children, we retrospectively examined medical records from 2008-2016 at a large Houston-area pediatric hospital and identified 36 cases. The earliest known cases were diagnosed in 2011.

Typhus Group Rickettsiosis, Texas, USA, 2003-2013.

We characterized the epidemiology of typhus group rickettsiosis in Texas, USA. During 2003-2013, a total of 1,762 cases were reported to the state health department. The number of diagnosed cases and geographic expansion increased over time. Physician awareness is critical to diagnose and effectively treat rickettsial infections.

Tip-link protein protocadherin 15 interacts with transmembrane channel-like proteins TMC1 and TMC2.

The tip link protein protocadherin 15 (PCDH15) is a central component of the mechanotransduction complex in auditory and vestibular hair cells. PCDH15 is hypothesized to relay external forces to the mechanically gated channel located near its cytoplasmic C terminus. How PCDH15 is coupled to the transduction machinery is not clear. Using a membrane-based two-hybrid screen to identify proteins that bind to PCDH15, we detected an interaction between zebrafish Pcdh15a and an N-terminal fragment of transmembrane channel-like 2a (Tmc2a). Tmc2a is an ortholog of mammalian TMC2, which along with TMC1 has been implicated in mechanotransduction in mammalian hair cells. Using the above-mentioned two-hybrid assay, we found that zebrafish Tmc1 and Tmc2a can interact with the CD1 or CD3 cytoplasmic domain isoforms of Pcdh15a, and this interaction depends on the common region shared between the two Pcdh15 isoforms. Moreover, an interaction between mouse PCDH15-CD3 and TMC1 or TMC2 was observed in both yeast two-hybrid assays and coimmunoprecipitation experiments. To determine whether the Pcdh15-Tmc interaction is relevant to mechanotransduction in vivo, we overexpressed N-terminal fragments of Tmc2a in zebrafish hair cells. Overexpression of the Tmc2a N terminus results in mislocalization of Pcdh15a within hair bundles, together with a significant decrease in mechanosensitive responses, suggesting that a Pcdh15a-Tmc complex is critical for mechanotransduction. Together, these results identify an evolutionarily conserved association between the fish and mouse orthologs of PCDH15 and TMC1 and TMC2, supporting the notion that TMCs are key components of the transduction complex in hair cells.

Health and Safety Threats to Ukraine From Nonconventional Weapons: A Clear and Present Danger.

This Viewpoint discusses the potential for use of nonconventional warfare threats (chemical, biological, radiological, nuclear, and explosives) in the conflict in Ukraine and how health care professionals need to recognize and respond to these threats.

Ebola virus outbreak, updates on current therapeutic strategies.

Filoviruses are enveloped negative-sense single-stranded RNA viruses, which include Ebola and Marburg viruses, known to cause hemorrhagic fever in humans with a case fatality of up to 90%. There have been several Ebola virus outbreaks since the first outbreak in the Democratic Republic of Congo in 1976 of which, the recent 2013-2015 epidemic in Guinea, Liberia, and Sierra Leone is the largest in recorded history. Within a few months of the start of the outbreak in December 2013, thousands of infected cases were reported with a significant number of deaths. As of March 2015, according to the Centers for Disease Control and Prevention, there have been nearly 25,000 suspected cases, with 15,000 confirmed by laboratory testing, and over 10,000 deaths. The large number of cases and the high mortality rate, combined with the lack of effective Food and Drug Administration-approved treatments, necessitate the development of potent and safe therapeutic measures to combat the current and future outbreaks. Since the beginning of the outbreak, there have been considerable efforts to develop and characterize protective measures including vaccines and antiviral small molecules, and some have proven effective in vitro and in animal models. Most recently, a cocktail of monoclonal antibodies has been shown to be highly effective in protecting non-human primates from Ebola virus infection. In this review, we will discuss what is known about the nature of the virus, phylogenetic classification, genomic organization and replication, disease transmission, and viral entry and highlight the current approaches and efforts, in the development of therapeutics, to control the outbreak.

Management of Burn Injuries in the Wilderness: Lessons from Low-Resource Settings.

Burns are a common source of injuries worldwide, with a high burden of disease in low- and middle-income countries. Burns also account for 2%-8% of wilderness injuries. Although many are minor, the potential for serious morbidity and mortality exists, and standard treatments used in high-resource settings are not readily available in the backcountry. A literature review was performed to find evidence from low-resource settings that supports alternative or improvised therapies that may be adapted to care of burns in the wilderness. There is good evidence for use of oral rehydration to support volume status in burn patients. There is moderate evidence to support cold therapy as first aid and adjunct for pain control. Some evidence supports use of alternative dressings such as boiled potato peel, banana leaf, aloe vera, honey, sugar paste, and papaya when standard therapies are not available.

Identification of sensory hair-cell transcripts by thiouracil-tagging in zebrafish.

BACKGROUND: Sensory hair cells are exquisitely sensitive to mechanical stimuli and as such, are prone to damage and apoptosis during dissections or in vitro manipulations. Thiouracil (TU)-tagging is a noninvasive method to label cell type-specific transcripts in an intact organism, thereby meeting the challenge of how to analyze gene expression in hair cells without the need to sort cells. We adapted TU-tagging to zebrafish to identify novel transcripts expressed in the sensory hair cells of the developing acoustico-lateralis organs. METHODS: We created a transgenic line of zebrafish expressing the T.gondii uracil phospho-ribosyltransferase (UPRT) enzyme specifically in the hair cells of the inner ear and lateral line organ. RNA was labeled by exposing 3 days post-fertilization (dpf) UPRT transgenic larvae to 2.5 mM 4-thiouracil (4TU) for 15 hours. Following total RNA isolation, poly(A) mRNA enrichment, and purification of TU-tagged RNA, deep sequencing was performed on the input and TU-tagged RNA samples. RESULTS: Analysis of the RNA sequencing data revealed the expression of 28 transcripts that were significantly enriched (adjusted p-value < 0.05) in the UPRT TU-tagged RNA relative to the input sample. Of the 25 TU-tagged transcripts with mammalian homologs, the expression of 18 had not been previously demonstrated in zebrafish hair cells. The hair cell-restricted expression for 17 of these transcripts was confirmed by whole mount mRNA in situ hybridization in 3 dpf larvae. CONCLUSIONS: The hair cell-restricted pattern of expression of these genes offers insight into the biology of this receptor cell type and may serve as useful markers to study the development and function of sensory hair cells. In addition, our study demonstrates the utility of TU-tagging to study nascent transcripts in specific cell types that are relatively rare in the context of the whole zebrafish larvae.