Mpox outbreak in Rio de Janeiro, Brazil: A translational approach.

Mpox is a zoonotic disease historically reported in Africa. Since 2003, limited outbreaks have occurred outside Africa. In 2022, the global spread of cases with sustained interhuman transmission and unusual disease features raised public health concerns. We explore the mpox outbreak in Rio de Janeiro (RJ) state, Brazil, in an observational study of mpox-suspected cases from June to December 2022. Data collection relied on a public healthcare notification form. Diagnosis was determined by MPXV-PCR. In 46 confirmed cases, anti-OPXV IgG was determined by ELISA, and seven MPXV genomes were sequenced. A total of 3095 cases were included, 816 (26.3%) with positive MPXV-PCR results. Most positive cases were men in their 30 s and MSM. A total of 285 (34.9%) MPXV-PCR+ patients live with HIV. Eight were coinfected with varicella-zoster virus. Anogenital lesions and adenomegaly were associated with the diagnosis of mpox. Females and individuals under 18 represented 9.4% and 5.4% of all confirmed cases, respectively, showing higher PCR cycle threshold (Ct) values and fewer anogenital lesions compared to adult men. Anti-OPXV IgG was detected in 29/46 (63.0%) patients. All analyzed sequences belonged to clade IIb. In RJ state, mpox presented a diverse clinical picture, represented mainly by mild cases with low complication rates and prominent genital involvement. The incidence in females and children was higher than usually reported. The observation of a bimodal distribution of Ct values, with few positive results, may suggest the need to review the diagnostic criteria in these groups.

Crack cocaine inhalation increases seizure susceptibility by reducing acetylcholinesterase activity.

Crack cocaine is a highly addictive and potent stimulant drug. Animal studies have shown that the cholinergic system plays a role in neurotoxicity induced by cocaine or its active metabolites inhalation. Behavioral alterations associated with crack cocaine use include hyperactivity, depressed mood, and decreased seizure threshold. Here we evaluate the acetylcholinesterase (AChE) and reactive oxygen species (ROS) activity, behavioral profile, and the threshold for epileptic seizures in rats that received intrahippocampal pilocarpine (H-PILO) followed by exposure to crack cocaine (H-PILO + CRACK). Animals exposed to H-PILO + CRACK demonstrated increased severity and frequency of limbic seizures. The AChE activity was reduced in the groups exposed to crack cocaine alone (CRACK) and H-PILO + CRACK, whereas levels of ROS remained unchanged. In addition, crack cocaine exposure increased vertical locomotor activity, without changing water and sucrose intake. Short-term memory consolidation remained unchanged after H-PILO, H-PILO + CRACK, and CRACK administration. Overall, our data suggest that crack cocaine inhalation reduced the threshold for epileptic seizures in rats submitted to low doses of pilocarpine through the inhibition of AChE. Taken together, our findings can be useful in the development of effective strategies for preventing and treating the harmful effects of cocaine and crack cocaine on the central nervous system.

Supplementation strategies for ewes during gestation and lactation.

Concentrated supplementation of ewes is a strategy to increase productivity. The objective was to evaluate the effects of supplementation in the diet of ewes before, during and in the final third of pregnancy and lactation on, the performance and production and composition of colostrum and milk. Forty animals were distributed in a completely randomized design, into the following treatments: CONT = control treatment with mineral salt supplementation only, SSREPRO = supplementation started 20 days before the estrus synchronization (ES) protocol, SSPREG = suplementação a partir da confirmação da gestação (60 days after ES), and SEPREG = supplementation in the third end of pregnancy (90 days after ES). Supplementation adoption changed (P < 0.05) the intake of organic matter, crude protein, neutral detergent fiber, the percentage of total digestible nutrients, and digestibility of dry matter, acid detergent fiber, non-fiber carbohydrates, and ether extract with their respective intake, in addition to colostrum and milk production and composition and animal performance. No difference (P > 0.05) was observed for organic matter, crude protein, and neutral detergent fiber digestibility and total digestible nutrient intake. Thus, supplementation in the final third of pregnancy may result in heavier lambs at weaning, reduction in slaughter time.

Melatonin improves the viability and ultrastructure of bovine oocyte-granulosa complexes of in vitro cultured early antral follicles.

This study aims to investigate the effects of melatonin on follicular growth, viability and ultrastructure, as well as on the levels of mRNA for antioxidant enzymes, reactive oxygen species (ROS) and meiotic progression in oocytes from in vitro cultured bovine early antral follicles. To this end, isolated early antral follicles (500-600 µm) were cultured in TCM-199+ alone or supplemented with 10-6 , 10-7 or 10-8 M melatonin at 38.5°C with 5% CO2 for 8 days. Follicle diameters were evaluated at days 0, 4 and 8 of culture. At the end of culture, ultrastructure, chromatin configuration, viability (calcein-AM and ethidium homodimer-1 staining), and the levels of ROS and mRNA for catalase (CAT), superoxide dismutase (SOD) and peroxiredoxin 6 (PRDX6) and glutathione peroxidase (GPx) were investigated in oocyte-granulosa cell complexes (OGCs). The results showed that early antral follicles cultured with 10-6 and 10-8 M melatonin had a progressive and significant increase in their diameters throughout the culture period (p < .05). Additionally, oocytes from follicles cultured with 10-7 or 10-8 M melatonin had increased fluorescence for calcein-AM, while those cultured with 10-6 or 10-7 M had reduced fluorescence for ethidium homodimer-1. Different from follicles cultured in other treatments, those cultured with 10-8 M melatonin had well-preserved ultrastructure of oocyte and granulosa cells. Melatonin, however, did not influence the levels of ROS, the mitochondrial activity, oocyte meiotic resumption and expression mRNA for SOD, CAT, GPX1 and PRDX6. In conclusion, the presence of 10-8 M melatonin in culture medium improves viability and preserves the ultrastructure of oocyte and granulosa cells of early antral follicles cultured in vitro.

A Survey of PAPR Techniques Based on Machine Learning.

Orthogonal Frequency Division Multiplexing (OFDM) is the modulation technology used in Fourth Generation (4G) and Fifth Generation (5G) wireless communication systems, and it will likely be essential to Sixth Generation (6G) wireless communication systems. However, OFDM introduces a high Peak to Average Power Ratio (PAPR) in the time domain due to constructive interference among multiple subcarriers, increasing the complexity and cost of the amplifiers and, consequently, the cost and complexity of 6G networks. Therefore, the development of new solutions to reduce the PAPR in OFDM systems is crucial to 6G networks. The application of Machine Learning (ML) has emerged as a promising avenue for tackling PAPR issues. Along this line, this paper presents a comprehensive review of PAPR optimization techniques with a focus on ML approaches. From this survey, it becomes clear that ML solutions offer customized optimization, effective search space navigation, and real-time adaptability. In light of the demands of evolving 6G networks, integration of ML is a necessity to propel advancements and meet increasing prerequisites. This integration not only presents possibilities for PAPR reduction but also calls for continued exploration to harness its potential and ensure efficient and reliable communication within 6G networks.

Influence of build direction, post-polymerization time, and aging on the fracture resistance and marginal and internal discrepancies of three-dimensional printed provisional crowns.

To evaluate the effect of build direction, post-polymerization time, and aging on the fracture resistance, failure mode, margin quality and Marginal (MD) and Internal Discrepancies (ID), and degree of conversion of tridimensional (3D) printed provisional crowns using the liquid crystal display technique (LCD). A left mandibular first molar was prepared and scanned. After, a full crown was designed (ExoCad) and exported to the LCD-printer software. One hundred and eighty crowns were printed at two build directions (30°, 90°) and post-polymerized for different periods (15, 30, 45 min). Half of the samples were aged (ST) in distilled water (37 °C/90 days). The marginal quality of the crowns was evaluated by the Schriwer method under a stereomicroscope (40x). The replica technique was used to measure the MD and ID and measured under a stereomicroscope (10x), through 20 reference points defined in 5 regions: Occlusal (O), Cusp (CP), Axial (AX), Chamfer (CH), and Finish Line (F). After, the crowns were cemented onto the respective dies using temporary cement and submitted to compression test (ISO150, 1 mm/min, 100 kgf).The failure mode was classified by Burke's fracture mode. The degree of conversion was evaluated through Fourier Transform Infrared Spectroscopy. The data (µm) of MD and ID and fracture resistance (N) were subjected to ANOVA (3 factors) and Tukey's test (5%); Weibull analysis was also performed for fracture resistance data (N). For MD, ID and fracture resistance ANOVA revealed that all factors (P < .001) were significant. For aged groups, the crowns printed at 30° showed lower MD (Tukey). The O_90_30 min (172.13A µm) and O_90_15 min (170.20A µm) groups showed the highest ID values. Higher resistance values were observed for the 30_45 min (844.30A N), 30_30 min (835.35A N), and 90_30 min (820.62A N) groups (Tukey). In the margin analysis, 98.6% of the crowns printed at 30° showed smooth margins without defects. The most prevalent fracture mode (41.7%) was Burke type 5. The degree of conversion (DC) increased with increasing post-polymerization time and aging. Printing provisional crowns at 30° provided lower crown MD and ID, as well as higher fracture resistance. A post-polymerization time of 30 min resulted in higher crown fracture resistance, while aging reduced the resistance of the crowns.

The oral health impact profile and well-being on mothers and preschool children.

OBJECTIVES: to verify the contribution of mothers' oral health impact profile to their children's oral health profile and the contribution of mothers' well-being and the caries index (dmft) to children's well-being. METHODS: This is a cross-sectional observational study. Mothers and pre-school children enrolled in public schools in the municipality of Araraquara-SP took part. The Oral Health Impact Profile Questionnaire (OHIP-14), Satisfaction with Life Scale (SWLS), Early Childhood Oral Health Impact Scale (ECOHIS) and the Autoquestionnaire Qualité de Vie Enfant Imagé (AUQEI) were used. To diagnose caries, a clinical examination was carried out to calculate the dmft index. Path analysis was carried out and the path coefficients were estimated (ß) and evaluated using the z-test (α = 5%). RESULTS: 443 children took part (5.19 ± 0.64 years; 52.4% boys) with an mean dmft of 1.31 ± 2.19. The mean age of the mothers was 33.4 ± 7.01 years. There was a significant impact of dmft and maternal well-being on the child's subjective well-being (s2explained = 43%). The mothers' oral health impact profile and the child's caries experience had a significant influence on both the child (OHIP: ß = 0.22; p < 0.001; dmft: ß = 0.48; p < 0.001) and the family (OHIP: ß = 0.29; p < 0.001; dmft: ß = 0.32; p < 0.001). The child's dmft (ß=-0.10; p = 0.005) and the mothers' subjective well-being (ß=-0.61; p < 0.001) had a significant impact on the child's subjective well-being. CONCLUSION: The mothers' oral health impact profile and the child's caries experience had an impact on both the child and the family. Mothers' subjective well-being and caries experience should be considered when assessing the subjective well-being of Brazilian preschool children.

Effect of different surface treatments and adhesive cementation on the surface topography and flexural strength of translucent and ultra-translucent monolithic zirconia.

PURPOSE: To evaluate the effect of different surface treatments and adhesive cementation on the miniflexural strength (MFS) of monolithic zirconia. MATERIALS AND METHODS: Two-hundred and forty (240) sintered bars of translucent zirconia (ZT) and ultra-translucent zirconia (ZUT) were obtained (8 mm ×2 mm ×1 mm). The bars were divided into 16 groups (n = 15) according to the factors "Zirconia" (ZT and ZUT), "Cementation" (Cem) and "surface treatment" (Ctrl:Control, Al:Aluminum oxide/Al2O3 50 µm, Si:Silica/SiO2 coated alumina particles oxide 30 µm, Gl:Glazing+hydrofluoric acid). Half of the bars received an adhesive layer application, followed by application of resin cement and light curing. The surface roughness was measured in non-cemented groups. All the bars were subjected to the MFS test (1.0 mm/min; 100 kgf). Scanning electron microscopy was used for qualitative analyses. MFS data (MPa) and roughness (µm) were statistically evaluated by three-way and two-way ANOVA respectively and Tukey's test (5%). RESULTS: The surface treatment and the interaction were significant for roughness. Glazing promoted less roughness compared to silicatization. Regarding MFS, only the zirconia and surface treatment factors were significant. For ZT, the sandblasted groups had an increase in MFS and glazing reduced it. There was no difference between the groups without cementation for the ZUT; however, ZUT.Si/Cem, and ZUT.Al/Cem obtained superior MFS among the cemented groups. CONCLUSIONS: Sandblasting increases the flexural strength for ZT, while glaze application tends to reduce it. Applying resin cement increases the flexural strength of ZUT when associated with sandblasting. Sandblasting protocols promote greater surface roughness.

Citrus essential oil: would it be feasible as antimicrobial in the bioethanol industry?

Essential oils (EOs) extracted from Citrus peels contain 85%-99% volatile components (a mixture of monoterpenes, sesquiterpenes, and their oxygenated derivatives) and 1%-15% non-volatile compounds. Citrus EOs have been long known for their antimicrobial properties, owing to which these EOs have a diverse range of applications. However, no studies have reported the applicability of Citrus EOs for the control of bacterial and yeast contaminants in the bioethanol industry. In this regard, the present review aimed to explore the feasibility of Citrus EOs in this industry. The Web of Science database was searched for reports that described the association of Citrus EOs with the most common microorganisms in the bioethanol industry to evaluate the efficacy of these EOs as antimicrobial agents in this context. The objective of the review was to suggest a novel antimicrobial that could replace sulfuric acid and antibiotics as the commonly used antimicrobial agents in the bioethanol industry. Citrus EOs exhibit antibacterial activity against Lactobacillus, which is the main bacterial genus that contaminates this fermentation process. The present report also confirms the selective action of these EOs on the contaminating yeasts and not/less on ethanol-producing yeast Saccharomyces cerevisiae, however further studies should be conducted to investigate the effects of Citrus EOs in yeast-bacterium co-culture.

Long-term effectiveness and safety of ustekinumab in bio-naïve and bio-experienced anti-tumor necrosis factor patients with Crohn's disease: a real-world multicenter Brazilian study.

BACKGROUND: The effectiveness of ustekinumab (UST) in the treatment of Crohn's disease (CD) has been demonstrated in the pivotal Phase 3 UNITI 1 and 2 and IM-UNITI studies in both anti-TNF-naïve and anti-TNF-exposed patients. Given the selective nature of pivotal trial designs, real-world effectiveness and safety studies are warranted. We report our experience with UST treatment in a large, real-world multicenter cohort of Brazilian patients with CD. METHODS: We performed a retrospective multicenter study including patients with CD, predominantly biologically refractory CD, who received UST. The primary endpoint was the proportion of patients in clinical remission at weeks 8, 24 and 56. Possible predictors of clinical and biological response/remission and safety outcomes were also assessed. RESULTS: Overall, 245 CD (mean age 39.9 [15-87]) patients were enrolled. Most patients (86.5%) had been previously exposed to biologics. According to nonresponder imputation analysis, the proportions of patients in clinical remission at weeks 8, 24 and 56 were 41.0% (n = 98/239), 64.0% (n = 153/239) and 39.3% (n = 94/239), respectively. A biological response was achieved in 55.4% of patients at week 8, and 59.3% were in steroid-free remission at the end of follow-up. No significant differences in either clinical or biological remission were noted between bio-naïve and bio-experienced patients. Forty-eight patients (19.6%) presented 60 adverse events during the follow-up, of which 8 (13.3%) were considered serious adverse events (3.2% of 245 patients). Overall, a proximal disease location, younger age, perianal involvement, and smoking were associated with lower rates of clinical remission over time. CONCLUSIONS: UST therapy was effective and safe in the long term in this large real-life cohort of Brazilian patients with refractory CD, regardless of previous exposure to other biological agents.

Per- and Polyfluoroalkyl Substances (PFAS) in Street Sweepings.

One hundred and seventeen street sweeping samples were collected and analyzed for per- and polyfluoroalkyl substances (PFAS). Fifty-six samples were collected in one city (Gainesville, Florida) allowing for an in-depth city-wide characterization. Street sweepings from five other urban areas, (Orlando, n = 15; Key West, n = 15; Pensacola, n = 12; Tampa, n = 13; and Daytona Beach, n = 6) were analyzed to provide a city-to-city comparison of PFAS. Within our analytical workflow, 37 PFAS were quantified across all samples, while the maximum number of PFAS quantified at one site was 26. Of those PFAS quantified in Gainesville, 60% were perfluoroalkyl acids (PFAAs) and 33% were precursors to PFAA. Among the PFAAs, short-chain perfluoroalkyl carboxylic acids (PFCAs) were the dominant class representing 26% of the total PFAS by concentration. In the comparison across different urban cities, the dominant compound by concentration and frequency of detection varied; however, perfluorooctanoic acid (PFOA) and linear perfluorooctanesulfonic acid (PFOSlin) were the two PFAS that were detected the most frequently. This study documents the first-time detection of hexadecafluorosebacic acid and perfluoro-3,6,9-trioxaundecane-1,11-dioic acid within environmental samples.

Biodegradation of the pyrethroid cypermethrin by bacterial consortia collected from orange crops.

Pyrethroids, such as cypermethrin (CYP), are widely employed in agriculture, promoting environmental pollution and the need for efficient decontamination methods. In this study, bacteria from orange crops were explored for CYP biodegradation. Among 40 tested bacterial strains, 20 grew in the presence of CYP and 19 performed statistically significant CYP biodegradation in 5 days (20.5%-97.8%). In addition, 3-phenoxybenzoic acid, the main metabolite from CYP, was quantified ranging from 1.1 mg.L-1 to 32.1 mg.L-1. The five most efficient strains, and consortia composed of 5, 10 and 20 bacteria biodegraded the CYP formulation as sole carbon source in phosphate buffer and in minimum mineral medium. Under optimized conditions determined employing Response Surface Methodology, Bacillus sp. CSA-1 and the consortium composed of 10 strains biodegraded 71.0% and 71.6% CYP in 24 h, respectively. Moreover, metabolite identification enabled the proposal of an extended biodegradation pathway with 29 identified compounds, including different new amide and amine derivatives that expanded the knowledge about the fate of this compound in the environment. Experiments of bioaugmentation in soil using Bacillus sp. CSA-1 and the consortium of 10 bacterial strains resulted in faster CYP biodegradation than natural attenuation, showing that the selection of efficient strains for composing a consortium is an interesting approach for bioremediation of pyrethroids.

Increased levels of perfluorooctanesulfonic acid (PFOS) during Hurricane Dorian on the east coast of Florida.

Per- and polyfluoroalkyl substances (PFAS) are a class of synthetic chemicals commonly found in everyday consumer products and are an emerging concern due to their ubiquitous presence in ecosystems around the world. PFAS exposure, which often occurs through contaminated water, has been linked to several adverse health effects in humans and wildlife. PFAS can be transported in surface water and storm runoff in the nearshore environment. Episodic events, such as hurricanes, are projected to increase in frequency and intensity, and a critical unanswered question is: how do episodic events influence the concentrations and distributions of emerging contaminants, such as PFAS, in coastal systems? Here, we investigated the impact of the 2019 Hurricane Dorian on the Florida coast to assess how natural disasters, such as hurricanes, influence the fate and transport of PFAS in surface water. Water samples collected throughout the St. Augustine Intracoastal waterway before, during, and after the storm were analyzed and compared with baseline concentrations. Ultra-high-pressure liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) was used in the detection and quantification of 23 and 17 PFAS, respectively. Perfluorooctane sulfonic acid (PFOS) was the compound with the highest concentration across all sampling sites. Mean PFOS levels showed the highest increase of 177% during the hurricane and returned to baseline levels after two days. Our findings highlight the need for continued research focused on understanding how large storms near all coastlines can impact the transport of environmental pollutants, such as PFOS, that can have adverse effects on human and environmental health. Further monitoring of PFAS in coastal systems is necessary to identify potential PFAS hotspots, investigate the impacts of episodic events on PFAS transport, develop mitigation practices capable of reducing the risk of PFAS exposure.

Blood parasites of passerines in the Brazilian Pampas and their implications for a potential population supplementation program for the endangered Yellow Cardinal (Gubernatrix cristata).

Espinilho savanna ("seasonal steppe savanna") is a unique vegetation formation of the Pampas biome that is found near the tri-border of Brazil, Uruguay, and Argentina. The Yellow Cardinal (Gubernatrix cristata) is a flagship species of this ecosystem, but it is classified as "critically endangered" in Brazil due to habitat loss and poaching for the illegal trade. Population supplementation through the release of individuals that were captive-bred or apprehended by authorities from the illegal trade has been considered as a conservation strategy for this species; however, the risk of pathogen introduction is a critical concern. We used microscopy and molecular methods to investigate the occurrence of blood parasites in wild passerines (n = 64, including three Yellow Cardinals) at Espinilho State Park, Rio Grande do Sul, Brazil, and in captive Yellow Cardinals (n = 30) at three facilities in Brazil. Haemosporidian parasites were detected in the blood smears of 10.9% of the wild passerines, comprising the morphospecies Haemoproteus erythrogravidus in Rufous-collared Sparrow (Zonotrichia capensis), H. quiscalus in Grayish Baywing (Agelaioides badius), and H. tyranni in Great Kiskadee (Pitangus sulphuratus); these are the southernmost records for these morphospecies and their first record for the Pampas biome. No haemosporidian parasites were detected in the blood smears of the Yellow Cardinals, wild or captive. Microfilariae were detected in the blood smears of 14.1% of the wild passerines, including all wild Yellow Cardinals, and in 43.3% of captive Yellow Cardinals. Trypanosoma sp. was detected in the blood smear of one captive Yellow Cardinal. Nested PCR and gene sequencing of the cyt-b gene of Haemoproteus/Plasmodium was used to test a subset of wild passerines and captive Yellow Cardinals, allowing for the molecular barcoding of H. quiscalus lineage AGEBAD04 and H. tyranni lineage PITSUL01; additionally, DNA identical to that of lineage PITSUL01 was detected in the blood of one captive Yellow Cardinal. This study provides valuable data to support the conservation management of the Yellow Cardinal and other threatened passerines from the Pampas and highlights the need for further studies on the epidemiology and pathology of filarioid worms and trypanosomes in passerines from this biome.

N-Acetyl cysteine reduces the levels of reactive oxygen species and improves in vitro maturation of oocytes from medium-sized bovine antral follicles.

This study aims to evaluate the effects of N-acetylcysteine (NAC) on bovine oocyte maturation, mitochondrial activity and transzonal projections (TZP), as well as on the levels of reactive oxygen species (ROS) and messenger RNA (mRNA) for catalase (CAT) superoxide dismutase (SOD), periredoxin-6 (Prdx6), glutathione peroxidase (GPx), growth and differentiation factor-9 (GDF9), histone H1Foo, cyclin B1 (CCNB1) and c-Mos. Bovine cumulus-oocyte complexes (COC) of medium-sized antral follicles (3.0-6.0 mm) were prematured in TCM-199 for 8 h at 38.5°C in 5% CO2. After prematuration in the presence of forskolin and C-type natriuretic peptide, COCs were matured in TCM-199 alone or with 0.1, 0.5 or 2.5 mM NAC. Then, oocytes were classified according to the stage of chromatin. Furthermore, mitochondrial activity and intracellular levels of ROS and TZP were also evaluated. The levels of mRNAs for CAT, SOD, Prdx6, GPx, GDF9, H1Foo, CCNB1 and c-Mos were evaluated using real-time polymerase chain reaction (RT-PCR). The results showed that NAC significantly increased the percentages of oocytes with resumption of meiosis when compared with those oocytes matured in control medium. Oocytes had homogeneous mitochondrial distribution, and those cultured with 0.1 and 0.5 mM NAC had lower levels of ROS when compared with the control. In addition, 0.5 mM NAC reduced TZP and the levels of mRNA for CCNB1. In contrast, NAC did not influence the expression of CAT, GPx, Prdx6, SOD, GDF9, H1Foo, and c-Mos. In conclusion, 0.5 mM NAC reduced the levels of ROS, TZP and mRNA for CCNB1, and improved in vitro resumption of meiosis in oocytes from medium-sized bovine antral follicles.

Differential Encoding of Predator Fear in the Ventromedial Hypothalamus and Periaqueductal Grey.

The ventromedial hypothalamus is a central node of the mammalian predator defense network. Stimulation of this structure in rodents and primates elicits abrupt defensive responses, including flight, freezing, sympathetic activation, and panic, while inhibition reduces defensive responses to predators. The major efferent target of the ventromedial hypothalamus is the dorsal periaqueductal gray (dPAG), and stimulation of this structure also elicits flight, freezing, and sympathetic activation. However, reversible inhibition experiments suggest that the ventromedial hypothalamus and periaqueductal gray play distinct roles in the control of defensive behavior, with the former proposed to encode an internal state necessary for the motivation of defensive responses, while the latter serves as a motor pattern initiator. Here, we used electrophysiological recordings of single units in behaving male mice exposed to a rat to investigate the encoding of predator fear in the dorsomedial division of the ventromedial hypothalamus (VMHdm) and the dPAG. Distinct correlates of threat intensity and motor responses were found in both structures, suggesting a distributed encoding of sensory and motor features in the medial hypothalamic-brainstem instinctive network.SIGNIFICANCE STATEMENT Although behavioral responses to predatory threat are essential for survival, the underlying neuronal circuits remain undefined. Using single unit in vivo electrophysiological recordings in mice, we have identified neuronal populations in the medial hypothalamus and brainstem that encode defensive responses to a rat predator. We found that both structures encode both sensory as well as motor aspects of the behavior although with different kinetics. Our findings provide a framework for understanding how innate sensory cues are processed to elicit adaptive behavioral responses to threat and will help to identify targets for the pharmacological modulation of related pathologic behaviors.

Benefits and challenges of nanomaterials in assisted reproductive technologies.

Assisted reproductive technology (ART) have contributed to preserve fertility in humans and to increase multiplication of genetically superior animals. Despite being highly practiced worldwide, ART presents some challenges, especially because gametes and embryos are kept in vitro for a variable period of time, and the oxidative stress in vitro can have negative impact on oocyte competence and embryo development. Nanotechnology needs to be considered to help overcome some of those impairments, since it can provide strategies to deliver antioxidants and hormones to gametes and embryos in vitro. The application of nanotechnology to ART can allow the development of new protocols using nanomaterials to improve in vitro oocyte competence and embryo production. This review discusses the applicability of nanomaterials to improve sperm selection, to deliver antioxidants and hormones to preantral follicles, oocytes, and embryos in vitro, as well as the concerns about using nanotechnology in ART.

Amygdala GluN2B-NMDAR dysfunction is critical in abnormal aggression of neurodevelopmental origin induced by St8sia2 deficiency.

Aggression is frequently observed in neurodevelopmental psychiatric disorders such as schizophrenia, autism, and bipolar disorder. Due to a lack of understanding of its underlying mechanisms, effective treatments for abnormal aggression are still missing. Recently, genetic variations in Sialyltransferase 2 (St8sia2) have been linked to these disorders and aggression. Here we identify abnormal aggressive behaviors and concomitant blunted fear learning in St8sia2 knockout (-/-) mice. It is worth noting that the amygdala of St8sia2-/- mice shows diminished threat-induced activation, as well as alterations in synaptic structure and function, including impaired GluN2B-containing NMDA receptor-mediated synaptic transmission and plasticity. Pharmacological rescue of NMDA receptor activity in the amygdala of St8sia2-/- mice with the partial agonist D-cycloserine restores synaptic plasticity and normalizes behavioral aberrations. Pathological aggression and associated traits were recapitulated by specific amygdala neonatal St8sia2 silencing. Our results establish a developmental link between St8sia2 deficiency and a pathological aggression syndrome, specify synaptic targets for therapeutic developments, and highlight D-cycloserine as a plausible treatment.

Microbiota identified from preserved Anopheles.

BACKGROUND: Mosquito species from the Anopheles gambiae complex and the Anopheles funestus group are dominant African malaria vectors. Mosquito microbiota play vital roles in physiology and vector competence. Recent research has focused on investigating the mosquito microbiota, especially in wild populations. Wild mosquitoes are preserved and transported to a laboratory for analyses. Thus far, microbial characterization post-preservation has been investigated in only Aedes vexans and Culex pipiens. Investigating the efficacy of cost-effective preservatives has also been limited to AllProtect reagent, ethanol and nucleic acid preservation buffer. This study characterized the microbiota of African Anopheles vectors: Anopheles arabiensis (member of the An. gambiae complex) and An. funestus (member of the An. funestus group), preserved on silica desiccant and RNAlater® solution. METHODS: Microbial composition and diversity were characterized using culture-dependent (midgut dissections, culturomics, MALDI-TOF MS) and culture-independent techniques (abdominal dissections, DNA extraction, next-generation sequencing) from laboratory (colonized) and field-collected mosquitoes. Colonized mosquitoes were either fresh (non-preserved) or preserved for 4 and 12 weeks on silica or in RNAlater®. Microbiota were also characterized from field-collected An. arabiensis preserved on silica for 8, 12 and 16 weeks. RESULTS: Elizabethkingia anophelis and Serratia oryzae were common between both vector species, while Enterobacter cloacae and Staphylococcus epidermidis were specific to females and males, respectively. Microbial diversity was not influenced by sex, condition (fresh or preserved), preservative, or preservation time-period; however, the type of bacterial identification technique affected all microbial diversity indices. CONCLUSIONS: This study broadly characterized the microbiota of An. arabiensis and An. funestus. Silica- and RNAlater®-preservation were appropriate when paired with culture-dependent and culture-independent techniques, respectively. These results broaden the selection of cost-effective methods available for handling vector samples for downstream microbial analyses.

Automated method to determine pharmaceutical compounds in wastewater using on-line solid-phase extraction coupled to LC-MS/MS.

An automated method was developed using on-line solid-phase extraction (SPE) as a sample preparation step, coupled to liquid chromatography-tandem mass spectrometry (LC-MS/MS), for determination of pharmaceutical compounds in wastewater samples at nanograms per liter to micrograms per liter concentration levels. This method is suitable for use in routine analysis, especially in environmental applications, avoiding cross-contamination and requiring minimal sample handling. Results can be obtained rapidly, with a chromatographic run time of only 24 min (including sample preparation and chromatographic analysis). Using a 50 µL injection volume, the method was validated according to international guidelines, considering parameters included in terms of method detection (MDL) and quantification limit (MQL), linearity, inter-day and intra-day precisions, and matrix effects. Assessment of chromatographic efficiency considered peak resolution and asymmetry, and carryover was evaluated to ensure analytical reliability and the ability to reuse the SPE cartridge. The intra- and inter-day precisions were lower than 10 and 17%, respectively. The MDL values ranged from 1×10-6 to 1 µg L-1, while the MQL values were from 0.001 to 3 µg L-1. Matrix effects were minimized by isotope dilution calibration. Application of the method to 20 wastewater samples showed that caffeine was the most frequently detected compound, with the highest concentration of 715 µg L-1, while other pharmaceutical compounds were detected in fewer samples and at lower concentrations (up to 8.51 µg L-1).