Freeze-Driven Synthesis of DNA Hairpin-Conjugated Gold Nanoparticle Biosensors for Dual-Mode Detection.

Freeze-based immobilization of deoxyribonucleic acid (DNA) oligonucleotides on gold nanoparticles (AuNPs) is highly efficient for single-stranded oligonucleotides but typically does not accommodate structures such as snap-cooled DNA hairpins (Sc-HPs) and snap-cooled molecular beacons (Sc-MBs) frequently used for biorecognition applications. Recognizing this limitation, we have developed a modified, freeze-based technique specifically designed to enable the adsorption of such hairpin oligonucleotides onto AuNP surfaces while ensuring that they retain their biosensing capabilities. Successful hairpin oligonucleotide conjugation of varying lengths to a wide range of AuNP diameters was corroborated by dynamic light scattering, ζ-potential, and UV-vis spectrophotometry. Moreover, we conducted a thorough evaluation of this modified method, confirming the retention of the sensing functions of Sc-HPs and Sc-MBs. This advancement not only offers a more efficient route for DNA hairpin conjugation but also elucidates the underlying biorecognition functions, with implications for broader applications in molecular diagnostics.

Add-on soft electronic interfaces for continuous cuffless blood pressure monitoring.

Continuous monitoring of arterial blood pressure is clinically important for the diagnosis and management of cardiovascular diseases. Soft electronic devices with skin-like properties show promise in a wide range of applications, including the human-machine interface, the Internet of things, and health monitoring. Here, we report the use of add-on soft electronic interfaces to address the connection challenges between soft electrodes and rigid data acquisition circuitry for bioimpedance monitoring of cardiac signals, including heart rate and cuffless blood pressure. Nanocomposite films in add-on electrodes provide robust electrical and mechanical contact with the skin and the rigid circuitry. We demonstrate bioimpedance sensors composed of add-on electrodes for continuous blood pressure monitoring with high accuracy. Specifically, the bioimpedance collected with add-on nanocomposite electrodes shows a signal-to-noise ratio of 37.0 dB, higher than the ratio of 25.9 dB obtained with standard silver/silver chloride (Ag/AgCl gel) electrodes. Although the sample set is low, the continuously measured systolic and diastolic blood pressure offer accuracy of -2.0 ± 6.3 mmHg and -4.3 ± 3.9 mmHg, respectively, confirming the grade A performance based on the IEEE standard. These results show promise in bioimpedance measurements with add-on soft electrodes for cuffless blood pressure monitoring.

Thermal History Mapping in Powder Bed Laser Sintering at the Micrometer Scale.

A thermal sensor was used to better understand parameters which influenced the interaction between a laser beam and a 0.5% Mn-doped ZnAl2.2O4 material, especially the laser defocusing parameter. The optical properties of the material depend on whether the Mn2+ ions occupy octahedral and/or tetrahedral sites depending on thermal history. A screen printing process is performed to obtain material thin films. Laser irradiation of the films was carried out (patterning 1 cm length single laser track) with different z defocusing heights. Luminescence properties around laser tracks led to the thermal history determination at the micrometer scale. It was shown that spatial thermal gradients defined at the micrometer scale perpendicularly to the borders of the laser tracks could be semiquantified for different z conditions. Laser defocusing leads to decrease thermal gradients as confirmed by thermal modeling studies.

Geometry design for a fully insertable glucose biosensor with multimodal optical readout.

Significance: Insertable optical continuous glucose monitors (CGMs) with wearable readers are a strong option for monitoring individuals with diabetes. However, a fully insertable CGM requires a small form factor while still delivering sufficient signal to be read through tissue by an external device. Previous work has suggested that a multimodal repeating unit (barcode) approach may meet these requirements, but the biosensor geometry must be optimized to meet performance criteria. Aim: This work details in silico trials conducted to evaluate the geometry of a fully insertable multimodal optical biosensor with respect to both optical output and species diffusion in vivo. Approach: Monte Carlo modeling is used to evaluate the luminescent output of three presupposed biosensor designs based on size constraints for an injectable and logical placement of the bar code compartments. Specifically, the sensitivity of the luminescent output to displacement of the biosensor in the X and Y directions, overall size of the selected design, and size of an individual repeating unit are analyzed. Further, an experimentally validated multiphysics model is used to evaluate the diffusion and reaction of glucose and oxygen within the biosensor to estimate the occurrence of chemical crosstalk between the assay components. Results: A stacked cylinder multimodal biosensor 4.4 mm in length with repeating units 0.36 mm in length was found to yield a greater luminescent output than the current "barcode" biosensor design. In addition, it was found that a biosensor with enzymatic elements does not significantly deplete glucose locally and thus does not impact the diffusion profile of glucose in adjacent compartments containing nonenzymatic assays. Conclusions: Computational modeling was used to design the geometry of a multimodal, insertable, and optical CGM to ensure that the optical output and chemical diffusion profile are sufficient for this device to function in vivo.

Detection of Hypoglycemia and Hyperglycemia Using Noninvasive Wearable Sensors: ECG and Accelerometry.

BACKGROUND: Monitoring glucose excursions is important in diabetes management. This can be achieved using continuous glucose monitors (CGMs). However, CGMs are expensive and invasive. Thus, alternative low-cost noninvasive wearable sensors capable of predicting glycemic excursions could be a game changer to manage diabetes. METHODS: In this article, we explore two noninvasive sensor modalities, electrocardiograms (ECGs) and accelerometers, collected on five healthy participants over two weeks, to predict both hypoglycemic and hyperglycemic excursions. We extract 29 features encompassing heart rate variability features from the ECG, and time- and frequency-domain features from the accelerometer. We evaluated two machine-learning approaches to predict glycemic excursions: a classification model and a regression model. RESULTS: The best model for both hypoglycemia and hyperglycemia detection was the regression model based on ECG and accelerometer data, yielding 76% sensitivity and specificity for hypoglycemia and 79% sensitivity and specificity for hyperglycemia. This had an improvement of 5% in sensitivity and specificity for both hypoglycemia and hyperglycemia when compared with using ECG data alone. CONCLUSIONS: Electrocardiogram is a promising alternative not only to detect hypoglycemia but also to predict hyperglycemia. Supplementing ECG data with contextual information from accelerometer data can improve glucose prediction.

Mid-Infrared Serial Microring Resonator Array for Real-Time Detection of Vapor-Phase Volatile Organic Compounds.

Chip-scale infrared spectrometers consisting of a microring resonator array (MRA) were developed for volatile organic compound (VOC) detection. The MRA is serially positioned to serve as a wavelength sorting element that enables wavelength demultiplexing. Unlike conventional devices operated by a single microring, our MRA can perform multiwavelength mid-infrared (mid-IR) sensing by routing the resonant wavelength light from a broadband mid-IR source into different sensing channels. Miniaturized spectrometer devices were fabricated on mid-IR transparent silicon-rich silicon nitride (SiNx) thin films through complementary metal-oxide-semiconductor (CMOS) processes, thus enabling wafer-level manufacturing and packaging. The spectral distribution of the resonance lines and the optimization of the microring structures were designed using finite-difference time-domain (FDTD) modeling and then verified by laser spectrum scanning. Using small microring structures, the spectrum showed a large free spectral range (FSR) of 100 nm and held four spectral channels without crosstalk. Unlike near-infrared microrings using refractive index sensing, our MRA can detect hexane and ethanol vapor pulses by monitoring the intensity variation at their characteristic mid-IR absorption bands, thus providing high specificity. Applying multiwavelength detection, the sensor module can discriminate among various VOC vapors. Hence, our mid-IR MRA could be an essential component to achieve a compact spectroscopic sensing module that has the potential for applications such as remote environmental monitoring and portable health care devices.

A Computational Modeling and Simulation Workflow to Investigate the Impact of Patient-Specific and Device Factors on Hemodynamic Measurements from Non-Invasive Photoplethysmography.

Cardiovascular disease is the leading cause of death globally. To provide continuous monitoring of blood pressure (BP), a parameter which has shown to improve health outcomes when monitored closely, many groups are trying to measure blood pressure via noninvasive photoplethysmography (PPG). However, the PPG waveform is subject to variation as a function of patient-specific and device factors and thus a platform to enable the evaluation of these factors on the PPG waveform and subsequent hemodynamic parameter prediction would enable device development. Here, we present a computational workflow that combines Monte Carlo modeling (MC), gaussian combination, and additive noise to create synthetic dataset of volar fingertip PPG waveforms representative of a diverse cohort. First, MC is used to determine PPG amplitude across age, skin tone, and device wavelength. Then, gaussian combination generates accurate PPG waveforms, and signal processing enables data filtration and feature extraction. We improve the limitations of current synthetic PPG frameworks by enabling inclusion of physiological and anatomical effects from body site, skin tone, and age. We then show how the datasets can be used to examine effects of device characteristics such as wavelength, analog to digital converter specifications, filtering method, and feature extraction. Lastly, we demonstrate the use of this framework to show the insensitivity of a support vector machine predictive algorithm compared to a neural network and bagged trees algorithm.

Dragon 1 Protocol Manuscript: Training, Accreditation, Implementation and Safety Evaluation of Portal and Hepatic Vein Embolization (PVE/HVE) to Accelerate Future Liver Remnant (FLR) Hypertrophy.

STUDY PURPOSE: The DRAGON 1 trial aims to assess training, implementation, safety and feasibility of combined portal- and hepatic-vein embolization (PVE/HVE) to accelerate future liver remnant (FLR) hypertrophy in patients with borderline resectable colorectal cancer liver metastases. METHODS: The DRAGON 1 trial is a worldwide multicenter prospective single arm trial. The primary endpoint is a composite of the safety of PVE/HVE, 90-day mortality, and one year accrual monitoring of each participating center. Secondary endpoints include: feasibility of resection, the used PVE and HVE techniques, FLR-hypertrophy, liver function (subset of centers), overall survival, and disease-free survival. All complications after the PVE/HVE procedure are documented. Liver volumes will be measured at week 1 and if applicable at week 3 and 6 after PVE/HVE and follow-up visits will be held at 1, 3, 6, and 12 months after the resection. RESULTS: Not applicable. CONCLUSION: DRAGON 1 is a prospective trial to assess the safety and feasibility of PVE/HVE. Participating study centers will be trained, and procedures standardized using Work Instructions (WI) to prepare for the DRAGON 2 randomized controlled trial. Outcomes should reveal the accrual potential of centers, safety profile of combined PVE/HVE and the effect of FLR-hypertrophy induction by PVE/HVE in patients with CRLM and a small FLR. TRIAL REGISTRATION: Clinicaltrials.gov: NCT04272931 (February 17, 2020). Toestingonline.nl: NL71535.068.19 (September 20, 2019).

Monte Carlo method for assessment of a multimodal insertable biosensor.

SIGNIFICANCE: Continuous glucose monitors (CGMs) are increasingly utilized as a way to provide healthcare to the over 10% of Americans that have diabetes. Fully insertable and optically transduced biosensors are poised to further improve CGMs by extending the device lifetime and reducing cost. However, optical modeling of light propagation in tissue is necessary to ascertain device performance. AIM: Monte Carlo modeling of photon transport through tissue was used to assess the luminescent output of a fully insertable glucose biosensor that uses a multimodal Förster resonance energy transfer competitive binding assay and a phosphorescence lifetime decay enzymatic assay. APPROACH: A Monte Carlo simulation framework of biosensor luminescence and tissue autofluorescence was built using MCmatlab. Simulations were first validated against previous research and then applied to predict the response of a biosensor in development. RESULTS: Our results suggest that a diode within the safety standards for light illumination on the skin, with far-red excitation, allows the luminescent biosensor to yield emission strong enough to be detectable by a common photodiode. CONCLUSIONS: The computational model showed that the expected fluorescent power output of a near-infrared light actuated barcode was five orders of magnitude greater than a visible spectrum excited counterpart biosensor.

Detection of volatile organic compounds using mid-infrared silicon nitride waveguide sensors.

Mid-infrared (mid-IR) sensors consisting of silicon nitride (SiN) waveguides were designed and tested to detect volatile organic compounds (VOCs). SiN thin films, prepared by low-pressure chemical vapor deposition (LPCVD), have a broad mid-IR transparent region and a lower refractive index (nSiN = 2.0) than conventional materials such as Si (nSi = 3.4), which leads to a stronger evanescent wave and therefore higher sensitivity, as confirmed by a finite-difference eigenmode (FDE) calculation. Further, in-situ monitoring of three VOCs (acetone, ethanol, and isoprene) was experimentally demonstrated through characteristic absorption measurements at wavelengths λ = 3.0-3.6 µm. The SiN waveguide showed a five-fold sensitivity improvement over the Si waveguide due to its stronger evanescent field. To our knowledge, this is the first time SiN waveguides are used to perform on-chip mid-IR spectral measurements for VOC detection. Thus, the developed waveguide sensor has the potential to be used as a compact device module capable of monitoring multiple gaseous analytes for health, agricultural and environmental applications.

A portable brightfield and fluorescence microscope toward automated malarial parasitemia quantification in thin blood smears.

Malaria is often most endemic in remote regions where diagnostic microscopy services are unavailable. In such regions, the use of rapid diagnostic tests fails to quantify parasitemia measurements which reflect the concentration of Plasmodium parasites in the bloodstream. Thus, novel diagnostic and monitoring technologies capable of providing such information could improve the quality of treatment, monitoring, and eradication efforts. A low-cost, portable microscope for gathering quantitative parasitemia data from fluorescently stained thin blood smears is presented. The system employs bimodal imaging using components optimized for cost savings, system robustness, and optical performance. The microscope is novel for its use of monochromatic visible illumination paired with a long working distance singlet aspheric objective lens that can image both traditionally mounted and cartridge-based blood smears. Eight dilutions of red blood cells containing laboratory cultured wild-type P. falciparum were used to create thin smears which were stained with SYBR Green-1 fluorescent dye. Two subsequent images are captured for each field-of-view, with brightfield images providing cell counts and fluorescence images providing parasite localization data. Results indicate the successful resolution of sub-micron sized parasites, and parasitemia measurements from the prototype microscope display linear correlation with measurements from a benchtop microscope with a limit of detection of 0.18 parasites per 100 red blood cells.

Skin optical properties in the obese and their relation to body mass index: a review.

SIGNIFICANCE: Obesity is a worldwide epidemic contributing directly to several cardiovascular risk factors including hypertension and type 2 diabetes. Wearable devices are becoming better at quantifying biomarkers relevant for the management of health and fitness. Unfortunately, both anecdotal evidence and recent studies indicate that some wearables have higher levels of error when utilized by populations with darker skin tones and high body mass index (BMI). There is an urgent need for a better evaluation of the limits of wearable health technologies when used by obese individuals. AIMS: (1) To review the current know-how on changes due to obesity in the skin epidermis, dermis, and subcutis that could affect the skin optical properties; (2) for the green wavelength range, to evaluate the difference in absorption and scattering coefficients from the abdominal skin between individuals with and without elevated BMI. The changes include alterations in layer thickness and cell size, as well as significant differences in chromophores and scatterer content, e.g., water, hemoglobin, collagen, and lipids. APPROACH: We have summarized literature pertaining to changes in skin and its components in obesity and report the results of our search using articles published between years 1971 and 2020. A linear model was used to demonstrate the absorption and reduced scattering coefficient of the abdominal skin of individuals with and without elevated BMI in the green wavelength range (530 to 550 nm) that is typically found in most wearables. RESULTS: The general trends indicate a decrease in absorption for both dermis and subcutis and an increase in reduced scattering for both epidermis and dermis. At 544-nm wavelength, a typical wavelength used for photoplethysmography (PPG), the absorption coefficient's relative percentage difference between high and low BMI skin, was 49% in the subcutis, 19% in the dermis, and negligible in the epidermis, whereas the reduced scattering coefficient relative difference was 21%, 29%, and 165% respectively. CONCLUSIONS: These findings suggest that there could be significant errors in the output of optical devices used for monitoring health and fitness if changes due to obesity are not accounted for in their design.

Spectrally multiplexed assay using gap enhanced nanoparticle for detection of a myocardial infarction biomarker panel.

Multiplexed assays are essential for the detection of biomarker panels. Differentiating signals from different biomarkers in a single test zone makes the detection more efficient. In this paper, a new method is designed for the synthesis of gap-enhanced nanoparticles (GeNPs) using Raman reporter molecules (RRM) and 6-amino-1-hexanethiol (6-AHT) as the spacer. The GeNPs show a nanometer-size gap, generate strong surface-enhanced Raman scattering (SERS) attributed to the gap, and exhibit discriminative spectral peaks. The strong Au-S bonds on both core and shell sides and the covalent bond between RRM and 6-AHT led to a stable structure, which ensured the stable SERS signal generation from the GeNPs. Using the GeNPs, a spectrally multiplexed assay for the detection of a biomarker panel is developed. The biomarker panel is composed of cardiac troponin I (cTnI), copeptin, and heart-type fatty acid-binding protein (h-FABP), which improves myocardial infarction (MI) diagnostic performance. A paper-based platform that is more amenable to point-of-care diagnostic analysis is used. The developed single biomarker assay achieves limits of detection of 0.01 ng mL-1, 0.86 ng mL-1, 0.004 ng mL-1 for cTnI, h-FABP, and copeptin in buffer solutions. The dynamic range of the assay in human serum samples also covers the clinically relevant range of the biomarkers. The cross interference in the multiplexed assay is low. These results show the strong potential of the developed GeNPs in multiplexed detection of biomarkers and the developed simple-to-use multiplexed assay in the diagnosis of MI at the point of care.

Cooperation between specialized livestock and crop farms can reduce environmental footprints and increase net profits in livestock production.

The rapid specialization of livestock production in China has contributed to spatially decoupled crop and livestock production, leading to various environmental pollution issues. Some regional agro-environmental policies have recently promoted the coupling of specialized crop and livestock farms through cooperation. However, the environmental and economic performances of such cooperation remain unclear. This study investigated multiple environmental footprints of two contrasting production systems: cooperative crop-livestock systems (CCLS) and decoupled specialized livestock systems (DSLS), using survey data of 87 ruminant farms in Northwest China. Results show that farms in CCLS had lower net greenhouse gas (GHG) emissions (12-29%), lower reactive nitrogen (Nr) emissions (21-40%), lower phosphorus footprints (PF) (41-54%), and used less cropland (24-31%) per kg animal product, compared to those in DSLS. The large differences in GHG emissions between the two systems were mainly related to enteric fermentation and resource production (used for feed production). The differences in Nr emissions and PF were mainly related to manure management. Net profits per kg animal product were higher in CCLS (13-35%) than in DSLS, and most profits originated from lower purchasing costs of feed and young livestock. Net profits and environmental footprints were negatively correlated, suggesting an environmental and economic win-win situation for CCLS. The possible obstacles to recoupling specialized crop and livestock farms through cooperation have been discussed, including farm size, contract stability, and local policies. Our study provides science-based evidence to support policymakers and specialized farms to close nutrient loops between crop and livestock production sectors through regional cooperation.

Predicting the Macronutrient Composition of Mixed Meals From Dietary Biomarkers in Blood.

Diet monitoring is an essential intervention component for a number of diseases, from type 2 diabetes to cardiovascular diseases. However, current methods for diet monitoring are burdensome and often inaccurate. In prior work, we showed that continuous glucose monitors (CGMs) may be used to predict meal macronutrients (e.g., carbohydrates, protein, fat) by analyzing the shape of the post-prandial glucose response. In this study, we examine a number of additional dietary biomarkers in blood by their ability to improve macronutrient prediction, compared to using CGMs alone. For this purpose, we conducted a nutritional study where (n = 10) participants consumed nine different mixed meals with varied but known macronutrient amounts, and we analyzed the concentration of 33 dietary biomarkers (including amino acids, insulin, triglycerides, and glucose) at various times post-prandially. Then, we built machine learning models to predict macronutrient amounts from (1) individual biomarkers and (2) their combinations. We find that the additional blood biomarkers provide complementary information, and more importantly, achieve lower normalized root mean squared error (NRMSE) for the three macronutrients (carbohydrates: 22.9%; protein: 23.4%; fat: 32.3%) than CGMs alone (carbohydrates: 28.9%, t(18) =1.64, p =0.060; protein: 46.4%, t(18) =5.38, p 0.001; fat: 40.0%, t(18) =2.09, p =0.025). Our main conclusion is that augmenting CGMs to measure these additional dietary biomarkers improves macronutrient prediction performance, and may ultimately lead to the development of automated methods to monitor nutritional intake. This work is significant to biomedical research as it provides a potential solution to the long-standing problem of diet monitoring, facilitating new interventions for a number of diseases.

A new species of Colobothea Lepeletier & Audinet-Serville, 1825 (Coleoptera, Cerambycidae, Lamiinae) from Peru, French Guiana, and Brazil, with notes on Colobothea hirtipes (Degeer, 1775).

Colobothea batesi sp. nov. is described based on specimens from Peru, French Guiana, and Brazil. Photographs of the male holotype and one female paratype are provided, as well as photographs of a male and a female of C. hirtipes (Degeer, 1775) to facilitate comparison. Notes on host plants of C. hirtipes are also provided.

Mongooses (Urva auropunctata) as reservoir hosts of Leptospira species in the United States Virgin Islands, 2019-2020.

During 2019-2020, the Virgin Islands Department of Health investigated potential animal reservoirs of Leptospira spp., the bacteria that cause leptospirosis. In this cross-sectional study, we investigated Leptospira spp. exposure and carriage in the small Indian mongoose (Urva auropunctata, syn: Herpestes auropunctatus), an invasive animal species. This study was conducted across the three main islands of the U.S. Virgin Islands (USVI), which are St. Croix, St. Thomas, and St. John. We used the microscopic agglutination test (MAT), fluorescent antibody test (FAT), real-time polymerase chain reaction (lipl32 rt-PCR), and bacterial culture to evaluate serum and kidney specimens and compared the sensitivity, specificity, positive predictive value, and negative predictive value of these laboratory methods. Mongooses (n = 274) were live-trapped at 31 field sites in ten regions across USVI and humanely euthanized for Leptospira spp. testing. Bacterial isolates were sequenced and evaluated for species and phylogenetic analysis using the ppk gene. Anti-Leptospira spp. antibodies were detected in 34% (87/256) of mongooses. Reactions were observed with the following serogroups: Sejroe, Icterohaemorrhagiae, Pyrogenes, Mini, Cynopteri, Australis, Hebdomadis, Autumnalis, Mankarso, Pomona, and Ballum. Of the kidney specimens examined, 5.8% (16/270) were FAT-positive, 10% (27/274) were culture-positive, and 12.4% (34/274) were positive by rt-PCR. Of the Leptospira spp. isolated from mongooses, 25 were L. borgpetersenii, one was L. interrogans, and one was L. kirschneri. Positive predictive values of FAT and rt-PCR testing for predicting successful isolation of Leptospira by culture were 88% and 65%, respectively. The isolation and identification of Leptospira spp. in mongooses highlights the potential role of mongooses as a wildlife reservoir of leptospirosis; mongooses could be a source of Leptospira spp. infections for other wildlife, domestic animals, and humans.

Postprandial concentration of circulating branched chain amino acids are able to predict the carbohydrate content of the ingested mixed meal.

BACKGROUND: The amount of the macronutrients protein and carbohydrate (CHO) in a mixed meal is known to affect each other's digestion, absorption, and subsequent metabolism. While the effect of the amount of dietary protein and fat on the glycemic response is well studied, the ability of postprandial plasma amino acid patterns to predict the meal composition is unknown. OBJECTIVE: To study the postprandial plasma amino acid patterns in relation to the protein, CHO, and fat content of different mixed meals and to investigate if these patterns can predict the macronutrient meal composition. DESIGN: Ten older adults were given 9 meals with 3 different levels (low, medium, and high) of protein, CHO, and fat in different combinations, taking the medium content as that of a standardized western meal. We monitored the postprandial plasma response for amino acids, glucose, insulin, and triglycerides for 8 h and the areas under the curve (AUC) were subsequently calculated. Multiple regression analysis was performed to determine if amino acid patterns could predict the meal composition. RESULTS: Increasing meal CHO content reduced the postprandial plasma response of several amino acids including all branched chain amino acids (BCAA) (leucine; q < 0.0001, isoleucine; q = 0.0035, valine; q = 0.0022). The plasma BCAA patterns after the meal significantly predicted the meal's CHO content (leucine; p < 0.0001, isoleucine; p = 0.0003, valine; p = 0.0008) along with aspartate (p < 0.0001), tyrosine (p < 0.0001), methionine (p = 0.0159) and phenylalanine (p = 0.0332). Plasma citrulline predicted best the fat content of the meal (p = 0.0024). CONCLUSIONS: The postprandial plasma BCAA patterns are lower with increasing meal CHO content and are strong predictors of a mixed meal protein and CHO composition, as are plasma citrulline for the fat content. We hypothesize that postprandial plasma amino acid concentrations can be used to predict the meal's macronutrient composition.

Determination of freedom-from-rabies for small Indian mongoose populations in the United States Virgin Islands, 2019-2020.

Mongooses, a nonnative species, are a known reservoir of rabies virus in the Caribbean region. A cross-sectional study of mongooses at 41 field sites on the US Virgin Islands of St. Croix, St. John, and St. Thomas captured 312 mongooses (32% capture rate). We determined the absence of rabies virus by antigen testing and rabies virus exposure by antibody testing in mongoose populations on all three islands. USVI is the first Caribbean state to determine freedom-from-rabies for its mongoose populations with a scientifically-led robust cross-sectional study. Ongoing surveillance activities will determine if other domestic and wildlife populations in USVI are rabies-free.

Simultaneous disruption of PRC2 and enhancer function underlies histone H3.3-K27M oncogenic activity in human hindbrain neural stem cells.

Driver mutations in genes encoding histone H3 proteins resulting in p.Lys27Met substitutions (H3-K27M) are frequent in pediatric midline brain tumors. However, the precise mechanisms by which H3-K27M causes tumor initiation remain unclear. Here, we use human hindbrain neural stem cells to model the consequences of H3.3-K27M on the epigenomic landscape in a relevant developmental context. Genome-wide mapping of epitope-tagged histone H3.3 revealed that both the wild type and the K27M mutant incorporate abundantly at pre-existing active enhancers and promoters, and to a lesser extent at Polycomb repressive complex 2 (PRC2)-bound regions. At active enhancers, H3.3-K27M leads to focal H3K27ac loss, decreased chromatin accessibility and reduced transcriptional expression of nearby neurodevelopmental genes. In addition, H3.3-K27M deposition at a subset of PRC2 target genes leads to increased PRC2 and PRC1 binding and augmented transcriptional repression that can be partially reversed by PRC2 inhibitors. Our work suggests that, rather than imposing de novo transcriptional circuits, H3.3-K27M drives tumorigenesis by locking initiating cells in their pre-existing, immature epigenomic state, via disruption of PRC2 and enhancer functions.