IFN-γ stimulates Paneth cell secretion through necroptosis mTORC1 dependent.

Immune mediators affect multiple biological functions of intestinal epithelial cells (IECs) and, like Paneth and Paneth-like cells, play an important role in intestinal epithelial homeostasis. IFN-γ a prototypical proinflammatory cytokine disrupts intestinal epithelial homeostasis. However, the mechanism underlying the process remains unknown. In this study, using in vivo and in vitro models we demonstrate that IFN-γ is spontaneously secreted in the small intestine. Furthermore, we observed that this cytokine stimulates mitochondrial activity, ROS production, and Paneth and Paneth-like cell secretion. Paneth and Paneth-like secretion downstream of IFN-γ, as identified here, is mTORC1 and necroptosis-dependent. Thus, our findings revealed that the pleiotropic function of IFN-γ also includes the regulation of Paneth cell function in the homeostatic gut.

First-in-Human PET Imaging of [18F]SDM-4MP3: A Cautionary Tale.

[18F]SynVesT-1 is a PET radiopharmaceutical that binds to the synaptic vesicle protein 2A (SV2A) and serves as a biomarker of synaptic density with widespread clinical research applications in psychiatry and neurodegeneration. The initial goal of this study was to concurrently conduct PET imaging studies with [18F]SynVesT-1 at our laboratories. However, the data in the first two human PET studies had anomalous biodistribution despite the injected product meeting all specifications during the prerelease quality control protocols. Further investigation, including imaging in rats as well as proton and carbon 2D-NMR spectroscopic studies, led to the discovery that a derivative of the precursor had been received from the manufacturer. Hence, we report our investigation and the first-in-human study of [18F]SDM-4MP3, a structural variant of [18F]SynVesT-1, which does not have the requisite characteristics as a PET radiopharmaceutical for imaging SV2A in the central nervous system.

Neuroinflammation After COVID-19 With Persistent Depressive and Cognitive Symptoms.

Importance: Persistent depressive symptoms, often accompanied by cognitive symptoms, commonly occur after COVID-19 illness (hereinafter termed COVID-DC, DC for depressive and/or cognitive symptoms). In patients with COVID-DC, gliosis, an inflammatory change, was suspected, but measurements of gliosis had not been studied in the brain for this condition. Objective: To determine whether translocator protein total distribution volume (TSPO VT), a marker of gliosis that is quantifiable with positron emission tomography (PET), is elevated in the dorsal putamen, ventral striatum, prefrontal cortex, anterior cingulate cortex, and hippocampus of persons with COVID-DC. Design, Setting, and Participants: This case-control study conducted at a tertiary care psychiatric hospital in Canada from April 1, 2021, to June 30, 2022, compared TSPO VT of specific brain regions in 20 participants with COVID-DC with that in 20 healthy controls. The TSPO VT was measured with fluorine F 18-labeled N-(2-(2-fluoroethoxy)benzyl)-N-(4-phenoxypyridin-3-yl)acetamide ([18F]FEPPA) PET. Main Outcomes and Measures: The TSPO VT was measured in the dorsal putamen, ventral striatum, prefrontal cortex, anterior cingulate cortex, and hippocampus. Symptoms were measured with neuropsychological and psychological tests, prioritizing outcomes related to striatal function. Results: The study population included 40 participants (mean [SD] age, 32.9 [12.3] years). The TSPO VT across the regions of interest was greater in persons with COVID-DC (mean [SD] age, 32.7 [11.4] years; 12 [60%] women) compared with healthy control participants (mean [SD] age, 33.3 [13.9] years; 11 [55%] women): mean (SD) difference, 1.51 (4.47); 95% CI, 0.04-2.98; 1.51 divided by 9.20 (17%). The difference was most prominent in the ventral striatum (mean [SD] difference, 1.97 [4.88]; 95% CI, 0.36-3.58; 1.97 divided by 8.87 [22%]) and dorsal putamen (mean difference, 1.70 [4.25]; 95% CI, 0.34-3.06; 1.70 divided by 8.37 [20%]). Motor speed on the finger-tapping test negatively correlated with dorsal putamen TSPO VT (r, -0.53; 95% CI, -0.79 to -0.09), and the 10 persons with the slowest speed among those with COVID-DC had higher dorsal putamen TSPO VT than healthy persons by 2.3 (2.30 divided by 8.37 [27%]; SD, 2.46; 95% CI, 0.92-3.68). Conclusions and Relevance: In this case-control study, TSPO VT was higher in patients with COVID-DC. Greater TSPO VT is evidence for an inflammatory change of elevated gliosis in the brain of an individual with COVID-DC. Gliosis may be consequent to inflammation, injury, or both, particularly in the ventral striatum and dorsal putamen, which may explain some persistent depressive and cognitive symptoms, including slowed motor speed, low motivation or energy, and anhedonia, after initially mild to moderate COVID-19 illness.

Programming the Cellular Uptake of Protein-Based Viromimetic Nanoparticles for Enhanced Delivery.

Viral mimetics is a noteworthy strategy to design efficient delivery systems without the safety drawbacks and engineering difficulties of modifying viral vectors. The triblock polypeptide CSB was previously designed de novo to self-assemble with DNA into nanocomplexes called artificial virus-like particles (AVLPs) due to their similarities to viral particles. Here, we show how we can incorporate new blocks into the CSB polypeptide to enhance its transfection without altering its self-assembly capabilities and the stability and morphology of the AVLPs. The addition of a short peptide (aurein) and/or a large protein (transferrin) to the AVLPs improved their internalization and specific targeting to cells by up to 11 times. Overall, these results show how we can further program the cellular uptake of the AVLPs with a wide range of bioactive blocks. This can pave the way to develop programmable and efficient gene delivery systems.

Multiplex gRNAs Synergically Enhance Detection of SARS-CoV-2 by CRISPR-Cas12a.

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) diagnostic methods have a large potential to effectively detect SARS-CoV-2 with sensitivity and specificity nearing 100%, comparable to quantitative polymerase chain reaction. Yet, there is room for improvement. Commonly, one guide CRISPR RNA (gRNA) is used to detect the virus DNA and activate Cas collateral activity, which cleaves a reporter probe. In this study, we demonstrated that using 2-3 gRNAs in parallel can create a synergistic effect, resulting in a 4.5 × faster cleaving rate of the probe and increased sensitivity compared to using individual gRNAs. The synergy is due to the simultaneous activation of CRISPR-Cas12a and the improved performance of each gRNA. This approach was able to detect as few as 10 viral copies of the N-gene of SARS-CoV-2 RNA after a preamplification step using reverse transcription loop-mediated isothermal amplification. The method was able to accurately detect 100% of positive and negative clinical samples in ∼25 min using a fluorescence plate reader and ∼45 min with lateral flow strips.

A case of facultative ophthalmomyiasis externa due to Calliphoridae and review of the literature.

Purpose: To describe a case of bilateral facultative ophthalmomyiasis externa due to Calliphoridae in a 30-year-old male assault victim at a suburban hospital in New York and review the relevant literature. Observations: An adult male was found to have maggot infestation of both eyes and severe secondary injury to the left cornea and ocular surface. He was treated with manual larvae removal, oral ivermectin, broad spectrum IV antibiotics, and topical antibiotics. Anterior segment reconstruction was required. Conclusions and importance: We report the first case of ophthalmomyiasis due to Calliphoridae in the United States and document the vision threatening potential of this rare condition. Timely examination by an ophthalmologist with early debridement may help prevent vision-threatening sequelae.

Dog hair volatiles attract Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) females.

Rhipicephalus sanguineus sensu lato (s.l.) (Latreille) feeds on domestic dogs worldwide. This tick species uses dog volatiles during host-seeking behavior. In this study, we identified volatile compounds from dog hairs involved in the host location of R. sanguineus s.l. The R. sanguineus s.l. females, but not males, were attracted to hair samples and Super Q extracts from Schnauzer dogs in Y-tube olfactometer bioassays. A total of 54 compounds from dog hair extracts were identified by gas chromatography coupled to mass spectrometry, including hydrocarbons, aldehydes, alcohols, ketones, and carboxylic acids. Screening the identified compounds by the single sensillum recording technique showed that isovaleric acid, hexanal, heptanal, and sucaltone (6-methyl-5-hepten-2-one) significantly stimulated the olfactory receptor neurons of the basiconic, chaeticum, and trichodeum sensilla of female ticks. When synthetic compounds were evaluated alone, or in binary, tertiary, or quaternary mixtures, female ticks were only attracted to isovaleric acid and 1 tertiary mixture (hexanal, heptanal, and isovaleric acid). We conclude that isovaleric acid functions as an attractant for R. sanguineus s.l. These findings contribute to the understanding of the chemical ecology of ticks during host location.

Extending resolution within a single imaging frame.

The resolution of fluorescence microscopy images is limited by the physical properties of light. In the last decade, numerous super-resolution microscopy (SRM) approaches have been proposed to deal with such hindrance. Here we present Mean-Shift Super Resolution (MSSR), a new SRM algorithm based on the Mean Shift theory, which extends spatial resolution of single fluorescence images beyond the diffraction limit of light. MSSR works on low and high fluorophore densities, is not limited by the architecture of the optical setup and is applicable to single images as well as temporal series. The theoretical limit of spatial resolution, based on optimized real-world imaging conditions and analysis of temporal image stacks, has been measured to be 40 nm. Furthermore, MSSR has denoising capabilities that outperform other SRM approaches. Along with its wide accessibility, MSSR is a powerful, flexible, and generic tool for multidimensional and live cell imaging applications.

Comparison of adjuvant emulsions for their safety and ability to enhance the antibody response in horses immunized with African snake venoms.

Adjuvant emulsions are widely used to enhance the antibody response in animals used as immunoglobulin source to produce snake antivenoms. We tested the performance of four commercial emulsion adjuvants (Montanide, Freund, Carbigen, and Emulsigen-D) and an experimental adjuvant (QH-769) in the antibody response of horses towards venoms of the African snakes Bitis arietans, Echis ocellatus, Dendroaspis polylepis and Naja nigricollis. Montanide, Freund and Carbigen adjuvants generated the highest immune response but induced moderate/severe local lesions at the site of injection. In contrast, Emulsigen-D and QH-769 adjuvants generated the lowest immune response and low incidence of local lesions. No evidence of systemic alterations was observed in the horses immunized with any of the adjuvants. It is suggested that the use of Montanide or Freund-based emulsions in the first immunization steps, followed by the use of Emulsigen-D, QH-769 or similar adjuvants in the following injections, could result in a satisfactory immune response against snake venoms, while not inducing serious local deleterious effects.

Effect of Polystyrene Synthesis Method on Water Sorption and Glass Transition.

Commodity PS is synthesized via free radical polymerization, whereas PS in block copolymers (BCPs) is typically synthesized via living anionic polymerization. The purpose of this work is to investigate how the synthesis method impacts important properties such as water sorption and glass transition temperature (Tg). Water sorption is important because the performance of nanostructured polymer membranes in various applications is known to be affected by environmental conditions such as humidity. Tg is important because it dictates processing conditions, both for commodity PS as well as BCPs such as thermoplastic elastomers. Water sorption in commercial PS was found to be 0.5 mgwater/gpolymer at the highest humidities investigated (about 80%), in agreement with literature. On the other hand, syndiotactic PS synthesized anionically at low temperature absorbed more water, up to 1.5 mgwater/gpolymer, due to higher free volume. The greatest impact on water sorption was due to addition of hydrophilic hydroxyl chain ends to atactic PS, which resulted in water sorption of up to 2.3 mgwater/gpolymer. In addition to measuring water sorption and dry Tg separately, the impact of relative humidity on PS Tg was examined. Combined differential scanning calorimetry and dynamic mechanical analysis show that on going from the dry state to high humidity, the Tg of PS decreases by 5 °C. Moreover, the tensile storage modulus of PS decreases from 1.58 GPa at 0% RH to 0.53 GPa at 40% RH. In addition to the practical relevance of this study, this report fills a gap in experimental literature by using a poor solvent system, PS/water, to examine plasticization in the pure polymer limit.

Endophytic fungus Phomopsis sp. as a source of 3-nitropropionic acid with larvicidal activity against Aedes aegypti (Linnaeus 1762, Diptera: Culicidae).

BACKGROUND: Aedes aegypti is the primary vector of viruses, such as Zika, chikungunya, yellow fever, and dengue. In this context, a biomonitored chemical study was conducted to evaluate the activity of the crude extract of the endophytic fungus Phomopsis sp. against the larvae of Aedes aegypti. METHODS: Crude extract, fractions, and isolated substances were evaluated in in-vitro assays against third-stage larvae of Aedes aegypti. RESULTS: We isolated 3-nitropropionic acid with an LC50 of 15.172 ppm and LC90 of 18.178 ppm after 24 hours of larval exposure. CONCLUSIONS: The results indicated that 3-nitropropionic acid exerted larvicidal activity.

La prevención de la fiebre del dengue, reto y desafío en la Atención Primaria de Salud / The prevention of dengue fever: a challenge in primary health care

RESUMEN Es indudable que, entre las arbovirosis, la fiebre del dengue constituye un grave problema de salud, por lo que se hace necesario incentivar a la población en aras de solucionar las dificultades que impiden la labor de prevención de esta enfermedad y el establecimiento de estrategias comunitarias con acciones educativas y de comunicación social por parte del médico y de la enfermera en la Atención Primaria de Salud, que permita consolidar el trabajo interdisciplinario e intersectorial en el enfrentamiento al vector y esta enfermedad como su consecuencia. El objetivo de la investigación es destacar la importancia del trabajo de prevención de esta enfermedad como reto y desafío en la Atención Primaria de Salud.

ABSTRACT There is no doubt that, among the arboviruses, dengue fever constitutes a serious health problem, for which it is necessary to encourage the population in order to solve the difficulties that impede the work of preventing this disease and the establishment of community strategies with educational and social communication actions by the doctor and the nurse in primary health care, which allows the consolidation of interdisciplinary and intersectoral work in confronting the vector and this disease as its consequence. The objective of the research is to highlight the importance of the work of prevention of this disease as a challenge in primary health care.

Diastereomeric Separation of Chiral fac-Tricarbonyl(iminopyridine) Rhenium(I) Complexes and Their Cytotoxicity Studies: Approach toward an Action Mechanism against Glioblastoma.

A series of new (tricarbonyl)rhenium(I) complexes were synthesized using chiral bidentate ligands (+)/(-)-iminopyridines (LR/LS). The reaction yielded a mixture of mononuclear Re(I) diastereoisomers, formulated as fac-[Br(CO)3Re(S/R)L(S/R)]. Each single diastereoisomer was isolated and fully characterized. X-ray crystallography and circular dichroism spectra verified their enantiomeric nature. The cytotoxicity of each complex was evaluated against six cancer cell lines. The effect of the two complexes on viability, proliferation, and migration was analyzed on glioblastoma cell lines (U251 and LN229). Changes in the expression of histones, apoptotic, and key signaling proteins, as well as alterations in DNA structure, were also observed. These experiments showed that the chirality associated with both metal and ligand has a strong influence on cytotoxicity.

Leveraging Citizen Science and Low-Cost Sensors to Characterize Air Pollution Exposure of Disadvantaged Communities in Southern California.

Assessing exposure to fine particulate matter (PM2.5) across disadvantaged communities is understudied, and the air monitoring network is inadequate. We leveraged emerging low-cost sensors (PurpleAir) and engaged community residents to develop a community-based monitoring program across disadvantaged communities (high proportions of low-income and minority populations) in Southern California. We recruited 22 households from 8 communities to measure residential outdoor PM2.5 concentrations from June 2021 to December 2021. We identified the spatial and temporal patterns of PM2.5 measurements as well as the relationship between the total PM2.5 measurements and diesel PM emissions. We found that communities with a higher percentage of Hispanic and African American population and higher rates of unemployment, poverty, and housing burden were exposed to higher PM2.5 concentrations. The average PM2.5 concentrations in winter (25.8 µg/m3) were much higher compared with the summer concentrations (12.4 µg/m3). We also identified valuable hour-of-day and day-of-week patterns among disadvantaged communities. Our results suggest that the built environment can be targeted to reduce the exposure disparity. Integrating low-cost sensors into a citizen-science-based air monitoring program has promising applications to resolve monitoring disparity and capture "hotspots" to inform emission control and urban planning policies, thus improving exposure assessment and promoting environmental justice.

Diagnostics of COVID-19 Based on CRISPR-Cas Coupled to Isothermal Amplification: A Comparative Analysis and Update.

The emergence of the COVID-19 pandemic prompted fast development of novel diagnostic methods of the etiologic virus SARS-CoV-2. Methods based on CRISPR-Cas systems have been particularly promising because they can achieve a similar sensitivity and specificity to the benchmark RT-qPCR, especially when coupled to an isothermal pre-amplification step. Furthermore, they have also solved inherent limitations of RT-qPCR that impede its decentralized use and deployment in the field, such as the need for expensive equipment, high cost per reaction, and delivery of results in hours, among others. In this review, we evaluate publicly available methods to detect SARS-CoV-2 that are based on CRISPR-Cas and isothermal amplification. We critically analyze the steps required to obtain a successful result from clinical samples and pinpoint key experimental conditions and parameters that could be optimized or modified to improve clinical and analytical outputs. The COVID outbreak has propelled intensive research in a short time, which is paving the way to develop effective and very promising CRISPR-Cas systems for the precise detection of SARS-CoV-2. This review could also serve as an introductory guide to new labs delving into this technology.

A kinetic model for the impact of packaging signal mimics on genome encapsulation.

Inspired by recent experiments on the spontaneous assembly of virus-like particles from a solution containing a synthetic coat protein and double-stranded DNA, we put forward a kinetic model that has as main ingredients a stochastic nucleation and a deterministic growth process. The efficiency and rate of DNA packaging strongly increase after tiling the DNA with CRISPR-Cas proteins at predesignated locations, mimicking assembly signals in viruses. Our model shows that treating these proteins as nucleation-inducing diffusion barriers is sufficient to explain the experimentally observed increase in encapsulation efficiency, but only if the nucleation rate is sufficiently high. We find an optimum in the encapsulation kinetics for conditions where the number of packaging signal mimics is equal to the number of nucleation events that can occur during the time required to fully encapsulate the DNA template, presuming that the nucleation events can only take place adjacent to a packaging signal. Our theory is in satisfactory agreement with the available experimental data.

Randomized comparison between perineural dexamethasone and combined perineural dexamethasone-dexmedetomidine for ultrasound-guided infraclavicular block.

BACKGROUND: This randomized trial compared perineural dexamethasone with combined perineural dexamethasone-dexmedetomidine for ultrasound-guided infraclavicular block. We hypothesized that the combination of perineural adjuvants would result in a longer motor block. METHODS: Fifty patients undergoing upper limb surgery with ultrasound-guided infraclavicular block (using 35 mL of lidocaine 1%-bupivacaine 0.25% with epinephrine 5 µg/mL) were randomly allocated to receive perineural dexamethasone (2 mg) or combined perineural dexamethasone (2 mg)-dexmedetomidine (50 µg). After the performance of the block, a blinded observer assessed the success rate (defined as a minimal sensorimotor composite score of 14 out of 16 points at 30 min), the onset time (defined as the time required to reach a minimal composite score of 14 points) as well as the incidence of surgical anesthesia (defined as the ability to complete surgery without local infiltration, supplemental blocks, intravenous opioids or general anesthesia).Postoperatively, the blinded observer contacted patients with successful blocks to inquire about the duration of motor block, sensory block and postoperative analgesia. RESULTS: No intergroup differences were observed in terms of success rate, onset time and surgical anesthesia. Compared with dexamethasone alone, combined dexamethasone-dexmedetomidine provided longer durations of motor block (21.5 (2.7) vs 17.0 (3.9) hours; p<0.001; 95% CI 2.6 to 6.4), sensory block (21.6 (3.6) vs 17.2 (3.6) hours; p<0.001; 95% CI 2.2 to 6.5), and postoperative analgesia (25.5 (9.4) vs 23.5 (5.6) hours; p=0.038; 95% CI 1.0 to 7.7). CONCLUSION: Compared with perineural dexamethasone (2 mg) alone, combined perineural dexamethasone (2 mg)-dexmedetomidine (50 µg) results in longer durations of sensorimotor block and analgesia. Further studies are required to determine the optimal dosing combination for dexamethasone-dexmedetomidine. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov identifier: NCT04875039.

Mites, rodents, and pathogens: A global review for a multi-species interaction in disease ecology.

More than ever, there is a need to understand how pathogens, vectors and hosts occur temporally and spatially to predict the occurrence of zoonotic outbreaks. Related to this, mites of the Mesostigmata and Trombidiformes orders have the potential to transmit several diseases, yet their information of occurrence, distribution and zoonotic accompanying agents have not been systematically organized. We conducted a systematic review using a combination of words through the Sysrev platform, as well as literature searches in specialised databases to identify global patterns of infections, trends in mite-rodent-pathogen research and existing knowledge gaps. The inclusion criterion was the detection of pathogens in mites, either by molecular or serological techniques or by direct observation under the microscope, which rendered 125 papers. Most works have been carried out in Asia, mainly around the transmission of Orientia tsutsugamushi. Interestingly, co-infections, i.e., the presence of more than one pathogen in an individual, are common in other Acari groups such as ticks. Moreover, this is not the case for Trombidiformes and Mesostigmata as only 4.7 percent of the articles reviewed detected more than one pathogen in these mites. It is important to include a multi-host, multi-vector and multi-pathogen approaches to understand complex systems in disease ecology. A synergy between mite taxonomists, physicians and veterinarians, decision-makers, governmental organisations, and society is needed to address the emergence of mite-borne new or neglected diseases.

Características clínicas y epidemiológicas de la estenosis hipertrófica del píloro en un hospital de Lima, Perú / Clinical and epidemiological characteristics of hypertrophic pyloric stenosis in a hospital in Lima, Perú

RESUMEN Objetivo: Determinar las características clínicas y epidemiológicas de la estenosis hipertrófica de píloro en un hospital de Lima, Perú. Materiales y métodos: Se realizó un estudio descriptivo, retrospectivo correspondiente a los años 2013 al 2019, en el servicio de Cirugía Pediátrica del Hospital Docente Madre Niño "San Bartolomé". Se recolectaron datos de las historias clínicas con diagnóstico de estenosis hipertrófica de píloro. Resultados: Participaron un total de 59 pacientes, de los cuales 79,7 % fueron varones, 68,4 % primogénitos, con una media de edad de 31.3 +/- 13.6 días de nacido. 100 % presentó vómitos, 44,1 % reptación abdominal y solo 35,6 % presentó oliva pilórica palpable. Tiempo quirúrgico promedio 51.39 minutos, estancia hospitalaria promedio de 6.35, se presentó complicaciones mayores en un 13,6 %, con una mortalidad de 3,4 %. Conclusiones: Es importante el inicio precoz de la vía oral, así como el manejo postquirúrgico en estos pacientes para disminuir su estancia hospitalaria y el desarrollo de complicaciones ventilatorias posquirúrgicas.

ABSTRACT Objective: To determine clinical and epidemiological characteristics of hypertrophic pyloric stenosis in a hospital in Lima, Peru. Materials and Methods: A descriptive and retrospective study encompassing years 2013 to 2019 was performed in the Pediatric Surgery Service of San Bartolome Teaching Mother and Child Hospital. Data from clinical records of patients diagnosed with hypertrophic pyloric stenosis was collected. Results: Records from fifty-nine patients were obtained, 79% were male, 68.4% were first born, and their mean age was 31.3 ± 13.6 days. All patients (100 %) had vomit, 44.1% had abnormal abdominal movement, and only 35.6% had palpable pyloric olive. The average surgical correction time was 51.39 minutes, and the average hospital stay was 6.35 days, major complications occurred in 13.6% of all children, and the mortality rate was 3.4%. Conclusions: Early initiation of oral intake is important, as well as post-surgical management in these patients, in order to reduce their in-hospital stay and the development of post-surgical ventilatory complications.