CBT+ Training Initiative in Washington State Community Mental Health: An Evaluation of Child Clinical Outcomes.

The Washington State CBT+ Initiative offers a flexible training and consultation approach for community mental health providers in evidence-based practices for four child mental health targets: cognitive behavioral therapy for depression, anxiety, trauma, and behavioral difficulties. As part of consultation, clinicians used an online system to track delivery of treatment components and clinical outcomes using standardized symptom measures. The current study used these clinician-input data to examine symptom change for children using paired sample t-tests. Additionally, we explored if time elapsed or number of sessions between measurements related to symptom change using simple linear regression. Children had significant symptom reduction across all four targets. For most measures, children did not show greater improvements with increased length of time or increased number of sessions between assessment measures. Findings suggest that children treated by a CBT+ trained clinician may demonstrate symptom reduction for their primary clinical problem. Findings add to support for flexible training approaches for community mental health clinicians.

Exploring the Effects of Rearing Densities on Epigenetic Modifications in the Zebrafish Gonads.

Rearing density directly impacts fish welfare, which, in turn, affects productivity in aquaculture. Previous studies have indicated that high-density rearing during sexual development in fish can induce stress, resulting in a tendency towards male-biased sex ratios in the populations. In recent years, research has defined the relevance of the interactions between the environment and epigenetics playing a key role in the final phenotype. However, the underlying epigenetic mechanisms of individuals exposed to confinement remain elucidated. By using zebrafish (Danio rerio), the DNA methylation promotor region and the gene expression patterns of six genes, namely dnmt1, cyp19a1a, dmrt1, cyp11c1, hsd17b1, and hsd11b2, involved in the DNA maintenance methylation, reproduction, and stress were assessed. Zebrafish larvae were subjected to two high-density conditions (9 and 66 fish/L) during two periods of overlapping sex differentiation of this species (7 to 18 and 18 to 45 days post-fertilization, dpf). Results showed a significant masculinization in the populations of fish subjected to high densities from 18 to 45 dpf. In adulthood, the dnmt1 gene was differentially hypomethylated in ovaries and its expression was significantly downregulated in the testes of fish exposed to high-density. Further, the cyp19a1a gene showed downregulation of gene expression in the ovaries of fish subjected to elevated density, as previously observed in other studies. We proposed dnmt1 as a potential testicular epimarker and the expression of ovarian cyp19a1a as a potential biomarker for predicting stress originated from high densities during the early stages of development. These findings highlight the importance of rearing densities by long-lasting effects in adulthood conveying cautions for stocking protocols in fish hatcheries.

Epigenetic variations are more substantial than genetic variations in rapid adaptation of oyster to Pacific oyster mortality syndrome.

Disease emergence is accelerating with global changes. Understanding by which mechanisms host populations can rapidly adapt will be crucial for management practices. Pacific oyster mortality syndrome (POMS) imposes a substantial and recurrent selective pressure on oyster populations, and rapid adaptation may arise through genetics and epigenetics. In this study, we used (epi)genome-wide association mapping to show that oysters differentially exposed to POMS displayed genetic and epigenetic signatures of selection. Consistent with higher resistance to POMS, the genes targeted included many genes in several pathways related to immunity. By combining correlation, DNA methylation quantitative trait loci, and variance partitioning, we revealed that a third of phenotypic variation was explained by interactions between the genetic and epigenetic information, ~14% by the genome, and up to 25% by the epigenome alone. Similar to genetically based adaptation, epigenetic mechanisms notably governing immune responses can contribute substantially to the rapid adaptation of hosts to emerging infectious diseases.

Streamlined pediatric anxiety program for school mental health services.

There needs to be serious transformation of evidence-based interventions (EBIs) into real-world solutions; otherwise, EBIs will never achieve the intended public health impact. In a randomized trial, we reported effects of a redesigned anxiety program. Herein, we described the redesign process that led to the program. Survey data revealed provider preferences for school mental health anxiety services. Focus groups and prototype feedback sessions revealed service barriers to uptake, implementation, and sustainability along with corresponding enabling strategies. Prototype feedback sessions also focused on refinement and fine-tuning of the redesign. In the end, traditional EBI strategies were transformed and packaged into six lessons, lasting 20-30 minutes each, and amenable to delivery in small-group format. The redesign achieved the intended purpose of retaining elements from cognitive and behavior therapy and social skills training for the target population of the intervention (e.g., 3rd to 5th graders with heterogeneous anxiety problems - identified and referred). The streamlined EBI is accessible from PBS LearningMedia™ - a service that hosts public, research-based, and school-ready materials.

Development of epigenetic biomarkers for the identification of sex and thermal stress in fish using DNA methylation analysis and machine learning procedures.

The sex ratio is a key ecological demographic parameter crucial for population viability. However, the epigenetic mechanisms operating during gonadal development regulating gene expression and the sex ratio remain poorly understood. Moreover, there is interest in the development of epigenetic markers associated with a particular phenotype or as sentinels of environmental effects. Here, we profiled DNA methylation and gene expression of 10 key genes related to sex development and stress, including steroidogenic enzymes, and growth and transcription factors. We provide novel information on the sex-related differences and on the influence of elevated temperature on these genes in zebrafish, a species with mixed genetic and environmental influences on sex ratios. We identified both positive (e.g., amh, cyp11c and hsd11b2) and negative (e.g., cyp11a1 and dmrt1) correlations in unexposed males, and negative correlation (amh) in exposed females between DNA methylation and gene expression levels. Further, we combined DNA methylation analysis with machine learning procedures and found a series of informative CpGs capable not only of correctly identifying sex (based on cyp19a1a DNA methylation levels) but also of identifying whether males and females had been exposed to abnormally elevated temperature when young (based on amh and foxl2a DNA methylation levels, respectively). This was achieved in the absence of conspicuous morphological alterations of the gonads. These DNA methylation-based epigenetic biomarkers represent molecular resources that can correctly recapitulate past thermal history and pave the way for similar findings in other species to assess potential ecological effects of environmental disturbances in the context of climate change.

A parasite outbreak in notothenioid fish in an Antarctic fjord.

Climate changes can promote disease outbreaks, but their nature and potential impacts in remote areas have received little attention. In a hot spot of biodiversity on the West Antarctic Peninsula, which faces among the fastest changing climates on Earth, we captured specimens of two notothenioid fish species affected by large skin tumors at an incidence never before observed in the Southern Ocean. Molecular and histopathological analyses revealed that X-cell parasitic alveolates, members of a genus we call Notoxcellia, are the etiological agent of these tumors. Parasite-specific molecular probes showed that xenomas remained within the skin but largely outgrew host cells in the dermis. We further observed that tumors induced neovascularization in underlying tissue and detrimentally affected host growth and condition. Although many knowledge gaps persist about X-cell disease, including its mode of transmission and life cycle, these findings reveal potentially active biotic threats to vulnerable Antarctic ecosystems.

Environmentally-induced sex reversal in fish with chromosomal vs. polygenic sex determination.

Sex ratio depends on sex determination mechanisms and is a key demographic parameter determining population viability and resilience to natural and anthropogenic stressors. There is increasing evidence that the environment can alter sex ratio even in genetically sex-determined species (GSD), as elevated temperature can cause female-to-male sex reversal (neomales). Alarmingly, neomales are being discovered in natural populations of several fish, amphibian and reptile species worldwide. Understanding the basis of neomale development is important for conservation biology. Among GSD species, it is unknown whether those with chromosomal sex determination (CSD), the most common system, will better resist the influence of high temperature than those with polygenic sex determination (PSD). Here, we compared the effects of elevated temperature in two wild zebrafish strains, Nadia (NA) and Ekkwill (EKW), which have CSD with a ZZ/ZW system, against the AB laboratory strain, which has PSD. First, we uncovered novel sex genotypes and the results showed that, at control temperature, the masculinization rate roughly doubled with the addition of each Z chromosome, while some ZW and WW fish of the wild strains became neomales. Surprisingly, we found that at elevated temperatures WW fish were just as likely as ZW fish to become neomales and that all strains were equally susceptible to masculinization. These results demonstrate that the Z chromosome is not essential for male development and that the dose of W buffers masculinization at the control temperature but not at elevated temperature. Furthermore, at the elevated temperature the testes of neomales, but not of normal males, contained more spermatozoa than at the control temperature. Our results show in an unprecedented way that, in a global warming scenario, CSD species may not necessarily be better protected against the masculinizing effect of elevated temperature than PSD species, and reveal genotype-by-temperature interactions in male sex determination and spermatogenesis.

Study of Molybdenite Floatability: Effect of Clays and Seawater.

Current challenges in froth flotation are the presence of complex gangues and the use of low-quality waters, such as seawater. In this scenario, the recovery of molybdenum minerals is difficult, mainly due to the hydrophobic faces' physicochemical changes. In the present study, the natural floatability of pure molybdenite was analyzed by using microflotation assays, and hydrophobicity was measured by performing contact-angle measurements. The impact of two clays, kaolin (non-swelling) and Na-montmorillonite (swelling), was studied. The behavior in freshwater and seawater at pH 8 was compared, considering the current condition of the Cu/Mo mining industries, which use seawater in their operations. The presence of clays lowered the natural floatability of molybdenite precisely because they adhere to the surface and reduce its contact angle. However, the intensity with which they cause this phenomenon depends on the type of water and clay. Kaolin strongly adheres to the valuable mineral in both freshwater and seawater. For its part, Na-montmorillonite does it with greater intensity in a saline medium, but in freshwater, a high concentration of phyllosilicate is required to reduce the hydrophobicity of molybdenite. The clays' adherence was validated by scanning electron microscopy (SEM) analysis.

A new insight into the restriction of Cr(VI) removal performance of activated carbon under neutral pH condition.

Activated carbon has been widely used to remove hazardous Cr(VI); however, the impact of Cr2O3 precipitate on gradually declining removal ability as pH increases has received little attention. Herein, to investigate the effect of Cr2O3, SEM-EDX (scanning electron microscope-energy dispersive X-ray analysis) coupling elements mapping of chromium-loaded powdered activated carbon (PAC) revealed that a chromium layer was formed on the PAC exterior after being treated with Cr(VI) at pH 7. XPS (X-ray photoelectron spectroscopy) study confirmed that 69.93% and 39.91% Cr2O3 precipitated on the PAC surface at pH 7 and pH 3, respectively, corresponding to 17.77 mg/g and 20 mg/g removal capacity. Exhausted PAC had a removal efficiency of 92.43% after Cr2O3 being washed by H2SO4 solution, which was much higher than the removal efficiency of 51.27 % after NaOH washing. This further verified that the intrinsically developed Cr2O3 precipitate on PAC under neutral conditions limited the durability of PAC as an adsorbent. Consecutive elution assessments confirmed that adsorption and reduction ability both declined as pH increased. Raman spectroscopy and C 1s spectra of materials demonstrated two distinct Cr(VI) removal mechanisms under pH 3 and pH 7. In conclusion, the exhausted AC after Cr(VI) adsorption can be rejuvenated after the surface coated Cr2O3 is washed by the acid solution, which can expand the longevity of AC and recover Cr(III).

Highly surface activated carbon to remove Cr(VI) from aqueous solution with adsorbent recycling.

To enhance the inferior removal capability of aqueous Cr(VI) by commercial activated carbon under neutral conditions. The emerging ball milling technology was employed and the removal efficiency of Cr(VI) by ball-milled highly activated carbon (HAC) increased from 68.3% to 99.0% under pH 6 and from 42.7% to 77.8% under pH 7 compared to pristine activated carbon (AC), respectively. Raman spectra and Boehm's titration results signified that the enhanced Cr(VI) removal performance of HAC under neutral conditions was associated with the enriched surface acid functional groups, in which the content of COOH groups increased from 0.31 mmol/g to 0.97 mmol/g. Two Cr(VI) removal mechanisms were proposed established on the acid and alkalic solution washed chromium-loaded HAC, involving the reduction of Cr(VI) to Cr(III) subsequently accompany with the formation of chromium hydroxides on the surface and inside the pores of HAC, and the bonding of CrO42- on the surface COOH groups, as confirmed by SEM-EDX element mapping and specific surface area and porosity measurements. The Pseudo-second order model and Freundlich model fitted the adsorption kinetic and isotherm of AC and HAC well severally, suggesting that the specific interaction of Cr(VI) with the HAC surface and the Cr(VI) removal was multi-layer adsorption. Thermodynamic study exhibited the spontaneity of Cr(VI) removal on ball-milled HAC was increased. Reusability and regeneration studies of HAC denoted the potential application on Cr(VI) uptake under neutral conditions.

Enhanced arsenic removal from water by a bimetallic material ZrOx-FeOx with high OH density.

Arsenic in groundwater for human consumption has negative effects on human's health worldwide. Due to the above, it is essential to invest in the development of new materials and more efficient technology for the elimination of such priority contaminants as arsenic. Therefore, in the present work, it was synthesized an amorphous hybrid material ZrOx-FeOx with a high density of OH groups, to improve the arsenic adsorption capacity of iron (FeOx) and zirconium (ZrOx) that makes up the bimetallic oxyhydroxide. The spectra of FT-IR and pKa's distribution suggest that in the synthesized binary oxides, a new union between the two metallic elements is formed by means of an oxygen (metal-O-metal). In addition, TEM profiles suggest that there are chemical interactions between both metals since no individual particles of iron oxide and zirconium oxide were found. According to the results, the adsorption capacity of the ZrOx-FeOx material increases 4.5 and 1.4 times with respect to FeOx and ZrOx, respectively. At pH 6, the maximum adsorption capacity was 27 mg g-1, but at pH greater than 7, the arsenic adsorption capacity onto ZrOx-FeOx decreased 66%. Graphical Abstract.

Exposure of zebrafish to elevated temperature induces sex ratio shifts and alterations in the testicular epigenome of unexposed offspring.

Accumulating evidence shows that environmental changes can affect population sex ratios through epigenetic regulation of gene expression in species where sex depends on both genetic and environmental cues. Sometimes, altered sex ratios persist in the next generation even when the environmental cue is no longer present (a multigenerational effect). However, evidence of transgenerational effects (i.e., beyond the first non-exposed generation), which tend to be paternally transmitted, is scarce and a matter of debate. Here, we used the AB strain of zebrafish, where sex depends on both genetic and environmental influences, to study possible multi- (to the F1) and transgenerational (to the F2) effects of elevated temperature during the critical period of sex differentiation. From eight initial different families, five were selected in order to capture sufficient variation between the sex ratio of the control group (28 °C) and the group exposed to elevated (35 °C) temperature only at the parental (P) generation. Results showed a consistent increase in the proportion of males in the P generation in all five families as a result of heat treatment. Sex ratios were then determined in the F1 and F2 offspring derived from both above groups, which were all raised at 28 °C. A persisting male-skewed sex ratio in the 35°C-derived, unexposed offspring of the F1 generation was observed in three families, denoting family-dependent multigenerational effects. However, no transgenerational effects were observed in the F2 generation of any family. DNA methylation was also assessed in the testis of P, F1 and F2 males derived from exposed and non-exposed fathers and grandfathers. DNA methylation was significantly decreased only in the testis of the 35°C-derived males in the F1 generation but not of the F2 generation and, surprisingly, neither in the 35°C-exposed males of the P generation. Taken together, our results show great interfamily variation, not only in sex ratio response to elevated temperature, but also on its multigenerational effects, denoting a strong influence of genetics. Alterations in the testicular epigenome in F1 males calls for attention to possible, previously unnoticed, effects of temperature in the unexposed offspring of heat-exposed parents in a global warming scenario.

The Model of the Conserved Epigenetic Regulation of Sex.

Epigenetics integrates genomic and environmental information to produce a given phenotype. Here, the model of Conserved Epigenetic Regulation of Sex (CERS) is discussed. This model is based on our knowledge on genes involved in sexual development and on epigenetic regulation of gene expression activation and silencing. This model was recently postulated to be applied to the sexual development of fish, and it states that epigenetic and gene expression patterns are more associated with the development of a particular gonadal phenotype, e.g., testis differentiation, rather than with the intrinsic or extrinsic causes that lead to the development of this phenotype. This requires the existence of genes with different epigenetic modifications, for example, changes in DNA methylation levels associated with the development of a particular sex. Focusing on DNA methylation, the identification of CpGs, the methylation of which is linked to sex, constitutes the basis for the identification of Essential Epigenetic Marks (EEM). EEMs are defined as the number and identity of informative epigenetic marks that are strictly necessary, albeit perhaps not sufficient, to bring about a specific, measurable, phenotype of interest. Here, we provide a summary of the genes where DNA methylation has been investigated so far, focusing on fish. We found that cyp19a1a and dmrt1, two key genes for ovary and testis development, respectively, consistently show an inverse relationship between their DNA methylation and expression levels, thus following CERS predictions. However, in foxl2a, a pro-female gene, and amh, a pro-male gene, such relationship is not clear. The available data of other genes related to sexual development such as sox9, gsdf, and amhr2 are also discussed. Next, we discuss the use of CERS to make testable predictions of how sex is epigenetically regulated and to better understand sexual development, as well as the use of EEMs as tools for the diagnosis and prognosis of sex. We argue that CERS can aid in focusing research on the epigenetic regulation of sexual development not only in fish but also in vertebrates in general, particularly in reptiles with temperature sex-determination, and can be the basis for possible practical applications including sex control in aquaculture and also in conservation biology.

Ovarian transcriptomic signatures of zebrafish females resistant to different environmental perturbations.

Sex is remarkably plastic in fish and can be easily influenced by environmental cues, in which temperature has been the most studied abiotic factor. However, it has been shown that elevated population densities can increase the number of males in several species but little is known about the underlying molecular mechanisms and whether general patterns exist. Here, we studied the long-term effects of population density on the gene expression program in zebrafish gonads. The ovarian transcriptome of females exposed to high versus low population densities contained 4,634 differentially expressed genes. Among them, a set of promale genes (amh, sypc3, spata6, and sox3) were upregulated in the high-population density group. Next, we compared the transcriptomes of ovaries of female zebrafish resistant to the masculinizing effects of either high density or elevated temperature. Results showed a set of 131 and 242 common upregulated and downregulated genes, respectively, including the upregulation of known male-related genes (e.g., amh and sycp3) but also genes involved in other functions (e.g., faima, ccm21, and ankrd6b) and a downregulation of cyp19a1a together with other genes (e.g., lgals9l1 and ubxn2a). We identified the common Gene Ontology terms involved in the reproduction and sexual development that were consistently affected in both environmental factors. These results show that regardless of the environmental perturbation there are common genes and cellular functions involved in the resistance to masculinization. These altered gene-expression profiles can be used as markers indicative of previous exposure to environmental stress independent of conspicuous alterations in sex ratios or gonadal morphology.

Preparation of microscale zero-valent iron-fly ash-bentonite composite and evaluation of its adsorption performance of crystal violet and methylene blue dyes.

New microscale zero-valent iron adsorbent on fly ash and bentonite matrix for removal of crystal violet (CV) and methylene blue (MB) was synthesized through direct reduction of iron oxide using coke and palm kernel shell. The adsorbent was prepared as cylindrical shaped pellets to remove the CV and MB from the aqueous solution. Nitrogen adsorption-desorption isotherm and scanning electron microscopy (SEM) studies showed that the adsorbent is highly porous, and the iron particles are finely dispersed on the supporting material surfaces. FTIR and UV studies indicated that the C=C bonds in CV and C=N+(CH3)2 bonds in MB were affected in the adsorption process. MB switched to the reduced MBH2 species while CV was reduced to two small-size molecular compounds, explaining the higher CV adsorption in comparison to that of MB. The reduction of these compounds was coupled to the oxidation of Fe0 to Fe2O3 as revealed by XRD characterization of the adsorbent after adsorption. CV and MB adsorption isotherms fitted well with the Langmuir adsorption model. Different adsorption and reduction kinetic models were examined for the MB and CV removal processes. A better fit of the experimental data with the pseudo-second-order model was observed. CV and MB adsorption increased with temperature in the 30-50 °C range. At 50 °C, adsorption capacities of CV and MB reached to 89.9 and 42.8 mg/g, respectively. This new adsorbent showed a superior adsorption capacity for CV and MB when compared to other adsorbents.

Appropriate rearing density in domesticated zebrafish to avoid masculinization: links with the stress response.

The zebrafish (Danio rerio) has become a well-established experimental model in many research fields but the loss of the primary sex-determining region during the process of domestication renders laboratory strains of zebrafish susceptible to the effects of environmental factors on sex ratios. Further, an essential husbandry aspect - the optimal rearing density to avoid stress-induced masculinization - is not known. We carried out two experiments: the first focusing on the effects of density on survival, growth and sex ratio by rearing zebrafish at different initial densities (9, 19, 37 and 74 fish per litre) for 3 months (6-90 days post-fertilization, dpf), and the second focusing on the effects of cortisol during the sex differentiation period (15-45 dpf) for zebrafish reared at low density. The results showed an increase in the number of males in groups subjected to the two highest initial rearing densities; we also observed a reduction of survival and growth in a density-dependent manner. Furthermore, zebrafish treated with cortisol during the sex differentiation period showed a complete masculinization of the population; treatment with the cortisol synthesis inhibitor metyrapone negated the effects of exogenous cortisol. Our results indicate that the process of sex differentiation in domesticated zebrafish can be perturbed by elevated stocking density and that this effect is likely to be mediated by an increase in cortisol through the stress response. However, the underlying mechanism needs further study.

Adsorption of As(V) inside the pores of porous hematite in water.

As(V) adsorption inside the pores of porous hematite in water has been studied in this work. This study was performed on nonporous hematite and porous hematite prepared from the thermal decomposition of goethite and siderite through the measurements of adsorption isotherm, SEM-EDX, XRD and BET. The results demonstrated that the As(V) adsorption was difficult to be realized inside pores if they were too small. This observation might be due to that the pore entrances were blocked by the adsorbed ions and thus the inside surfaces became invalid for the adsorption. Only if the pore size is large enough, the effective surface area inside pores would be close to that on non-porous hematite for As(V) adsorption. In addition, it was found that siderite is better than goethite for preparing porous hematite with thermal decomposition as adsorbent for arsenic removal.

Effects of oxidation on the defect of reduced graphene oxides in graphene preparation.

The relationship between the defects of reduced graphene oxide (RGO) and the oxidation degree of graphite in the preparation of graphene with chemical conversion method has been studied in this work. This study was performed on an artificial graphite through the measurements of X ray diffraction, Raman spectroscopy and particle size analysis. The experimental results have shown that there indeed was a close relationship between the defects and the oxidation degree, which appeared in the form of S-type curve. Also, it was found that a low KMnO4 addition would lead to a partial oxidation of graphite, leaving defects mainly on the edges of RGO; with a high KMnO4 addition, the defects on RGO mostly appeared on the surfaces.