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Christianne E Strang, Mary Katherine Ray, Mary M Boggiano, Franklin R Amthor Department of Psychology, The University of Alabama at Birmingham, Birmingham, AL, USA Purpose: Transcranial direct current stimulation (tDCS) has been studied in humans for its effects on enhancement of learning, amelioration of psychiatric disorders, and modification of other behaviors for over 50 years. Typical treatments involve injecting 2 mA current through scalp electrodes for 20 minutes, sometimes repeated weekly for two to five sessions. Little is known about the direct effects of tDCS at the neural circuit or the cellular level. This study assessed the effects of tDCS-like currents on the central nervous system by recording effects on retinal ganglion cell responsiveness using the rabbit retina eyecup preparation. Materials and methods: We examined changes in firing to On and Off light stimuli during and after brief applications of a range of currents and polarity and in different classes of ganglion cells. Results: The responses of Sustained cells were consistently suppressed during the first round of current application, but responses could be enhanced after subsequent rounds of stimulation. The observed first round suppression was independent of current polarity, amplitude, or number of trials. However, the light responses of Transient cells were more likely to be enhanced by negative currents and unaffected or suppressed by first round positive currents. Short-duration currents, that is, minutes, as low as 2.5 µA produced a remarkable persistency of firing changes, for up to 1.5 hours, after cessation of current. Conclusion: The results are consistent with postulated tDCS alteration of central nervous system function, which outlast the tDCS session and provide evidence for the isolated retina as a useful model to understand tDCS actions at the neuronal level. Keywords: tDCS mechanisms, CNS, in vitro model, neural coding, neuromodulation

Female jail populations are comprised of women at highrisk for an array of psychological and physical health problems. Jails offer an opportune site to deliver clinical health interventions to women who often quickly cycle back into the community. In contrast with prison population studies, many investigators have en-countered recruitment problems when attempting to engage the jailed population in clinical research. This study addressed the feasibility of recruiting detained women for eligibility for clinical research.Methods: Commitments to the Women's Facility at the Rhode Island Department of Corrections were chronicled for 40 months, from February 2004 to June 2007. Research staff, working 8 a.m. to 5 p.m., Monday through Friday, attempted to screen all detained women for a randomized clinical trial.Results: During the 40-month study period, 4,131 individual women had 8,010 commitments to the facility. Staff was able to gain access to nearly 50% of women. Of the inaccessible women, 65% were released in less than 24 hours. In total, 88% of accessed women agreed to be screened for study participation. No significant differences were observed by race/ethnicity or age between women who were screened and those who were not.Conclusions: Clinical research with the female jail population is feasible. The jail setting requires researchers to plan for short-commitment lengths and high rates of recidivism to optimize screening and recruitment in this population. [ABSTRACT FROM AUTHOR]

RNase R (encoded by the rnr gene) is a highly processive 3' → 5' exoribonuclease essential for the growth of the psychrotrophic bacterium Pseudomonas syringae Lz4W at low temperature. The cell death of a rnr deletion mutant at low temperature has been previously attributed to processing defects in 16S rRNA, defective ribosomal assembly, and inefficient protein synthesis. We recently showed that RNase R is required to protect P. syringae Lz4W from DNA damage and oxidative stress, independent of its exoribonuclease activity. Here, we show that the processing defect in 16S rRNA does not cause cell death of the rnr mutant of P. syringae at low temperature. Our results demonstrate that the rnr mutant of P. syringae Lz4W, complemented with a RNase R deficient in exoribonuclease function (RNase R D284A ), is defective in 16S rRNA processing but can grow at 4 °C. This suggested that the processing defect in ribosomal RNAs is not a cause of the cold sensitivity of the rnr mutant. We further show that the rnr mutant accumulates copies of the indigenous plasmid pLz4W that bears a type II toxin-antitoxin (TA) system (P. syringae antitoxin-P. syringae toxin). This phenotype was rescued by overexpressing antitoxin psA in the rnr mutant, suggesting that activation of the type II TA system leads to cold sensitivity of the rnr mutant of P. syringae Lz4W. Here, we report a previously unknown functional relationship between the cold sensitivity of the rnr mutant and a type II TA system in P. syringae Lz4W., Competing Interests: Conflict of interest The authors have no conflict of interest with the contents of this article., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The study aimed to identify common differentially expressed lncRNAs from manually curated ulcerative colitis (UC) gene expression omnibus (GEO) datasets. Nine UC transcriptomic datasets of clearly annotated human colonic biopsies were included in the study. The datasets were manually curated to select active UC samples and controls. R packages geneknitR, gprofiler, clusterProfiler were used for gene symbol annotation. The R EdgeR package was used to analyze differential expression. This resulted in a total of nineteen lncRNAs that were differentially expressed in at least three datasets of the nine GEO datasets. Several of the differentially expressed lncRNAs found in UC were associated with promoting colorectal cancer (CRC) through regulating gene expression, epithelial to mesenchymal transition (EMT), cell cycle progression, and by promoting tumor proliferation, invasion, and migration. The expression of several lncRNAs varied between disease states and tissue locations within the same disease state. The identified differentially expressed lncRNAs may function as general markers for active UC independent of biopsy location, age, gender, or treatment, thereby representing a comparative resource for future comparisons using available GEO UC datasets.

The development of humanized monoclonal antibodies (MAbs) with Lutetium-177 ([ 177 Lu]Lu 3+ ) has brought a paradigm shift in the arena of targeted therapy of various cancers. [ 177 Lu]Lu-DOTA-Rituximab and [ 177 Lu]Lu-DOTA-Trastuzumab have gained prominence due to their improved therapeutic efficacy in the treatment of lymphoma and breast cancer. The clinical dose formulation of these radiolabeled MAbs, using low specific activity [ 177 Lu]LuCl 3 , requires extensive optimization of the radiolabeling protocol. The present study merits the development of a single protocol which has been optimized for conjugation of Rituximab and Trastuzumab with p-NCS-benzyl-DOTA and further radiolabeling these immunoconjugates (ICs) with low specific activity [ 177 Lu]LuCl 3 . Herein, we report a consistent and reproducible protocol for clinical dose formulations of [ 177 Lu]Lu-DOTA-Rituximab and [ 177 Lu]Lu-DOTA-Trastuzumab (~9.25 GBq each, equivalent to ~2 patient doses) with radiochemical yield (RCY) between 84 and 86% and radiochemical purities (RCP) >99%. The in vitro stabilities of both these radioimmunoconjugates (RICs) were retained up to 120 h post-radiolabeling, upon storage with L-ascorbic acid as stabilizer (concentration: ~ 220-240 μg/37MBq) at -20 °C. The ready-to-use formulation of clinical doses[ 177 Lu]Lu-DOTA-Rituximab and [ 177 Lu]Lu-DOTA-Trastuzumab has been successfully achieved by employing a single optimized protocol. While [ 177 Lu]Lu-DOTA-Rituximab has exhibited a high degree of localization in retroperitoneal nodal mass of refractory lymphoma patient, high uptake of [ 177 Lu]Lu-DOTA-Trastuzumab has been observed in metastatic breast carcinoma patient with multiple skeletal metastases., (© 2023. The Author(s), under exclusive licence to World Molecular Imaging Society.)

Importance: RNA metabolism is important as RNA acts as a link between genomic information and functional biomolecules, thereby playing a critical role in cellular response to environment. We investigated the role of DEAD-box RNA helicases in low-temperature adapted growth of P. syringae , as this group of enzymes play an essential role in modulation of RNA secondary structures. This is the first report on the assessment of all major DEAD-box RNA helicases in any Antarctic bacterium. Of the five RNA helicases, three ( srmB , csdA , and dbpA ) are important for the growth of the Antarctic P. syringae at low temperature. However, the requisite role of dbpA and the indispensable requirement of csdA for low-temperature adapted growth are a novel finding of this study. Growth analysis of combinatorial deletion strains was performed to understand the functional interaction among helicase genes. Similarly, genetic complementation of RNA helicase mutants was conducted for identification of gene redundancy in P. syringae ., Competing Interests: The authors declare no conflict of interest.

Objectives: The present work describes the automated radiochemical synthesis of different PET tracers like [ 68 Ga]Ga-Pentixafor, [ 68 Ga]Ga-FAPI-4 and [ 68 Ga]Ga-DOTATATE using optimized single protocol in the non-cassette based Eckert & Ziegler (EZ) Modular Lab (fixed tubing system) without any modification in the inbuilt human machine interface (HMI) software. Recently, PET agents viz. [ 68 Ga]Ga-Pentixafor and [ 68 Ga]Ga-FAPI-4 are gaining prominence for the diagnosis of overexpressed Chemokine Receptor-4 (CXCR4) and Fibroblast Activation Protein (FAP) receptor, respectively, in the microenvironment of numerous cancer types. The promising results observed with the clinical usage of [ 68 Ga]Ga-DOTATATE produced using the automated protocol, provided impetus for the clinical translation of [ 68 Ga]Ga-Pentixafor and [ 68 Ga]Ga-FAPI-4 using the in-house developed automated radiolabeling protocol., Methods: Herein we report a single radiolabeling protocol for the automated preparation of [ 68 Ga]Ga-Pentixafor and [ 68 Ga]Ga-FAPI-4 in the non-cassette based EZ Modular-Lab Standard radiochemistry module, without any changes in schematic, graphical user interface (GUI) software and time list, from that used for routine production of [ 68 Ga]Ga-DOTATATE in our centre, since 2015. Physico-chemical quality control and in-vitro stability analyses were carried out using radio-TLC and radio-HPLC., Results: The automated protocol yielded reliable and consistent non-decay corrected (ndc) radiochemical yield (RCY) of (84.4%±0.9%) and (85.5%±1.4%) respectively, for [ 68 Ga]Ga-Pentixafor and [ 68 Ga]Ga-FAPI-4, with RCP>98%, which are comparable to the RCY of (84.4%±1.2%) and RCP (99.1%±0.3%) for [ 68 Ga]Ga-DOTATATE. The biological quality control studies confirmed the formulations to be of ready-to-use pharmaceutical grade., Conclusion: The consistent and reliable RCY and RCP of multiple 68 Ga-labeled PET tracers by single automated radiochemistry protocol exhibits the versatility of the EZ Modular Lab., (© 2024 mums.ac.ir All rights reserved.)

Importance: Bacterial exoribonucleases play a crucial role in RNA maturation, degradation, quality control, and turnover. In this study, we have uncovered a previously unknown role of 3'-5' exoribonuclease RNase R of Pseudomonas syringae Lz4W in DNA damage and oxidative stress response. Here, we show that neither the exoribonuclease function of RNase R nor its association with the RNA degradosome complex is essential for this function. Interestingly, in P. syringae Lz4W, hydrolytic RNase R exhibits physiological roles similar to phosphorolytic 3'-5' exoribonuclease PNPase of E. coli . Our data suggest that during the course of evolution, mesophilic E. coli and psychrotrophic P. syringae have apparently swapped these exoribonucleases to adapt to their respective environmental growth conditions., Competing Interests: The authors declare no conflict of interest.

Background: In Antarctic P. syringae RNase R play an essential role in the processing of 16S and 5S rRNA, thereby playing an important role in cold-adapted growth of the bacterium. This study is focused on deciphering the in vivo functional activity of mesophilic exoribonuclease R and its catalytic domain (RNB) in an evolutionary distant psychrophilic bacterium Pseudomonas syringae Lz4W., Results: Our results confirm that E. coli RNase R complemented the physiological functions of the psychrophilic bacterium P. syringae RNase R and rescued the cold-sensitive phenotype of Pseudomonas syringae ∆rnr mutant. More importantly, the catalytic domain (RNB) of the E. coli RNase R is also capable of alleviating the cold-sensitive growth defects of ∆rnr mutant as seen with the catalytic domain (RNB) of the P. syringae enzyme. The Catalytic domain of E. coli RNase R was less efficient than the Catalytic domain of P. syringae RNase R in rescuing the cold-sensitive growth of ∆rnr mutant at 4°C, as the ∆rnr expressing the RNB Ec (catalytic domain of E. coli RNase R) displayed longer lag phase than the RNB Ps (Catalytic domain of P. syringae RNase R) complemented ∆rnr mutant at 4°C. Altogether it appears that the E. coli RNase R and P. syringae RNase R are functionally exchangeable for the growth requirements of P. syringae at low temperature (4°C). Our results also confirm that in P. syringae the requirement of RNase R for supporting the growth at 4°C is independent of the degradosomal complex., Conclusion: E. coli RNase R (RNase R Ec ) rescues the cold-sensitive phenotype of the P. syringae Δrnr mutant. Similarly, the catalytic domain of E. coli RNase R (RNB Ec ) is also capable of supporting the growth of Δrnr mutant at low temperatures. These findings have a vast scope in the design and development of low-temperature-based expression systems., (© 2023. Academy of Scientific Research and Technology.)

Long non-coding RNAs (lncRNAs) have been shown to play a role in the pathogenesis of ulcerative colitis (UC). Although epigenetic processes such as DNA methylation and lncRNA expression are well studied in UC, the importance of the interplay between the two processes has not yet been fully explored. It is, therefore, believed that interactions between environmental factors and epigenetics contribute to disease development. Mucosal biopsies from 11 treatment-naïve UC patients and 13 normal controls were used in this study. From each individual sample, both whole-genome bisulfite sequencing data (WGBS) and lncRNA expression data were analyzed. Correlation analysis between lncRNA expression and upstream differentially methylated regions (DMRs) was used to identify lncRNAs that might be regulated by DMRs. Furthermore, proximal protein-coding genes associated with DMR-regulated lncRNAs were identified by correlating their expression. The study identified UC-associated lncRNAs such as MIR4435-2HG, ZFAS1, IL6-AS1, and Pvt1, which may be regulated by DMRs. Several genes that are involved in inflammatory immune responses were found downstream of DMR-regulated lncRNAs, including SERPINB1, CCL18, and SLC15A4. The interplay between lncRNA expression regulated by DNA methylation in UC might improve our understanding of UC pathogenesis.

This paper presents a robust colon cancer diagnosis method based on the feature selection method. The proposed method for colon disease diagnosis can be divided into three steps. In the first step, the images' features were extracted based on the convolutional neural network. Squeezenet, Resnet-50, AlexNet, and GoogleNet were used for the convolutional neural network. The extracted features are huge, and the number of features cannot be appropriate for training the system. For this reason, the metaheuristic method is used in the second step to reduce the number of features. This research uses the grasshopper optimization algorithm to select the best features from the feature data. Finally, using machine learning methods, colon disease diagnosis was found to be accurate and successful. Two classification methods are applied for the evaluation of the proposed method. These methods include the decision tree and the support vector machine. The sensitivity, specificity, accuracy, and F1Score have been used to evaluate the proposed method. For Squeezenet based on the support vector machine, we obtained results of 99.34%, 99.41%, 99.12%, 98.91% and 98.94% for sensitivity, specificity, accuracy, precision, and F1Score, respectively. In the end, we compared the suggested recognition method's performance to the performances of other methods, including 9-layer CNN, random forest, 7-layer CNN, and DropBlock. We demonstrated that our solution outperformed the others.

Neuroinflammation is both a consequence and driver of overfeeding and weight gain in rodent obesity models. Advances in magnetic resonance imaging (MRI) enable investigations of brain microstructure that suggests neuroinflammation in human obesity. To assess the convergent validity across MRI techniques and extend previous findings, we used diffusion basis spectrum imaging (DBSI) to characterize obesity-associated alterations in brain microstructure in 601 children (age 9-11 years) from the Adolescent Brain Cognitive Development SM Study. Compared with children with normal-weight, greater DBSI restricted fraction (RF), reflecting neuroinflammation-related cellularity, was seen in widespread white matter in children with overweight and obesity. Greater DBSI-RF in hypothalamus, caudate nucleus, putamen, and, in particular, nucleus accumbens, correlated with higher baseline body mass index and related anthropometrics. Comparable findings were seen in the striatum with a previously reported restriction spectrum imaging (RSI) model. Gain in waist circumference over 1 and 2 years related, at nominal significance, to greater baseline RSI-assessed restricted diffusion in nucleus accumbens and caudate nucleus, and DBSI-RF in hypothalamus, respectively. Here we demonstrate that childhood obesity is associated with microstructural alterations in white matter, hypothalamus, and striatum. Our results also support the reproducibility, across MRI methods, of findings of obesity-related putative neuroinflammation in children., (© The Author(s) 2023. Published by Oxford University Press.)

Importance: Lifestyle change is central to diabetes risk reduction in youth with overweight or obesity. Feeling susceptible to a health threat can be motivational in adults., Objective: To evaluate associations between diabetes risk perception and/or awareness and health behaviors in youth., Design, Setting, and Participants: This cross-sectional study analyzed data from the US National Health and Nutrition Examination Survey 2011 to 2018. Participants included youths aged 12 to 17 years with body mass index (BMI) in the 85th percentile or higher without known diabetes. Analyses were conducted from February 2022 to February 2023., Main Outcomes and Measures: Outcomes included physical activity, screen time, and attempted weight loss. Confounders included age, sex, race and ethnicity, and objective diabetes risk (BMI, hemoglobin A1c [HbA1c])., Exposures: Independent variables included diabetes risk perception (feeling at risk) and awareness (told by clinician), as well as potential barriers (eg, food insecurity, household size, insurance)., Results: The sample included 1341 individuals representing 8 716 794 US youths aged 12 to 17 years with BMI in the 85th percentile or higher for age and sex. The mean age was 15.0 years (95% CI, 14.9-15.2 years) and mean BMI z score was 1.76 (95% CI 1.73-1.79). Elevated HbA1c was present in 8.6% (HbA1c 5.7%-6.4%: 8.3% [95% CI, 6.5%-10.5%]; HbA1c ≥6.5%: 0.3% [95% CI, 0.1%-0.7%]). Nearly one-third of youth with elevated HbA1c reported risk perception (30.1% [95% CI, 23.1%-38.1%), while one-quarter (26.5% [95% CI, 20.0%-34.2%]) had risk awareness. Risk perception was associated with increased TV watching (β = 0.3 hours per day [95% CI, 0.2-0.5 hours per day]) and approximately 1 less day per week with at least 60 minutes of physical activity (β = -1.2 [95% CI, -2.0 to -0.4) but not with nutrition or weight loss attempts. Awareness was not associated with health behaviors. Potential barriers had mixed associations: larger households (≥5 members vs 1-2) reported lower consumption of non-home-prepared meals (OR 0.4 [95% CI, 0.2-0.7]) and lower screen time (β = -1.1 hours per day [95% CI, -2.0 to -0.3 hours per day), while public insurance (vs private) was associated with approximately 20 fewer minutes per day of physical activity (β = -20.7 minutes per day [95% CI, 35.5 to -5.8 minutes per day])., Conclusions and Relevance: In this cross-sectional study including a US-representative sample of adolescents with overweight or obesity, diabetes risk perception and awareness were not associated with greater engagement in risk-reducing behaviors in youth. These findings suggest the need to address barriers to engagement in lifestyle change, including economic disadvantage.

Background: The clinical profile and course of COVID-19 evolved perilously in a second wave, leading to the use of various treatment modalities that included homeopathy. This prognostic factor research (PFR) study aimed to identify clinically useful homeopathic medicines in this second wave., Methods: This was a retrospective, multi-centred observational study performed from March 2021 to May 2021 on confirmed COVID-19 cases who were either in home isolation or at COVID Care Centres in Delhi, India. The data were collected from integrated COVID Care Centres where homeopathic medicines were prescribed along with conventional treatment. Only those cases that met a set of selection criteria were considered for analysis. The likelihood ratio (LR) was calculated for the frequently occurring symptoms of the prescribed medicines. An LR of 1.3 or greater was considered meaningful., Results: Out of 769 confirmed COVID-19 cases reported, 514 cases were selected for analysis, including 467 in home isolation. The most common complaints were cough, fever, myalgia, sore throat, loss of taste and/or smell, and anxiety. Most cases improved and there was no adverse reaction. Certain new symptoms, e.g., headache, dryness of mouth and conjunctivitis, were also seen. Thirty-nine medicines were prescribed, the most frequent being Bryonia alba followed by Arsenicum album , Pulsatilla nigricans , Belladonna , Gelsemium sempervirens , Hepar sulphuris , Phosphorus , Rhus toxicodendron and Mercurius solubilis . By calculating LR, the prescribing indications of these nine medicines were ascertained., Conclusion: Add-on use of homeopathic medicines has shown encouraging results in the second wave of COVID-19 in integrated care facilities. Further COVID-related research is required to be undertaken on the most commonly prescribed medicines., Competing Interests: None declared., (Faculty of Homeopathy. This article is published by Thieme.)

Background:  Most of the symptoms of coronavirus disease 2019 (COVID-19) are covered by large repertory rubrics and hence many remedies have been proposed as "genus epidemicus". The aim of this study was to combine the information from various data collections to prepare a COVID-19 Bayesian mini-repertory/an algorithm-based application (app) and test it., Methods:  In July 2021, 1,161 COVID-19 cases from 100 practitioners globally were combined. These data were used to calculate "condition-confined" likelihood ratios (LRs) for 59 symptoms of COVID-19. Out of these, 35 symptoms of the 11 medicines that had at least 20 cases each were considered. The information was entered in a spreadsheet (algorithm) to calculate combined LRs of specific combinations of symptoms. The algorithm contained the medicines Arsenicum album , Belladonna , Bryonia alba , Camphora , Gelsemium sempervirens , Hepar sulphuris , Mercurius solubilis , Nux vomica , Phosphorus , Pulsatilla and Rhus toxicodendron . To test concordance, the doctors were then invited to re-enter the symptoms of their cases into this algorithm., Results:  The algorithm was re-tested on 358 cases, and concordance was seen in 288 cases. On analysis of the data, bias was noticed in the Merc group, which was therefore excluded from the algorithm. The remaining 10 medicines, representing 81.8% of all cases, were included in the preparation of the next version of the homeopathic mini-repertory and app., Conclusion:  The Bayesian mini-repertory and app is based on qualitative clinical experiences of various doctors in COVID-19 and gives indications for specific medicines for common COVID-19 symptoms. It is freely available [English: https://hpra.co.uk/; Spanish: https://hpra.co.uk/es ] for further testing and utilization by the profession., Competing Interests: None declared., (Faculty of Homeopathy. This article is published by Thieme.)

DEAD box RNA helicases are involved in important cellular processes like RNA metabolism (Processing and Degradation), ribosome biogenesis and translation. Besides being crucial to the formation of cold adapted degradosomes, RNA helicases have been implicated in structural rearrangement of RNA, implying a role in alleviation of RNA secondary structure stabilization at low temperature. This study depicts the results of experiments on protective role played by DEAD box RNA helicases against nucleic acid damaging agents. RNA helicase mutants ΔrhlE, ΔsrmB, ΔcsdA, ΔdbpA and ΔrhlB cells were exposed to various DNA damaging agents (UV, Paraquat, Mitomycin C, Hydroxyurea and Hydrogen peroxide) and assessed for sensitivity to them. Our results illustrate that ∆csdA displayed sensitivity to paraquat (that causes oxidative damage) and UV radiation induced DNA damage. On the other hand, ∆srmB displays sensitivity to hydroxyurea that causes damage to the replication forks (RFs) by inhibiting ribonucleotide reductase and depleting the dNTP pool of cells. However, all five RNA helicase mutants were resistant to H 2 O 2 mediated oxidative stress and mitomycin C induced DNA cross-links., Competing Interests: Declaration of Competing Interest There are no competing interests to declare., (Copyright © 2022. Published by Elsevier B.V.)

The widespread use of fertilizers is a result of the increased global demand for food. The commonly used chemical fertilizers may increase plant growth and output, but they have deleterious effects on the soil, the environment, and even human health. Therefore, nanofertilizers are one of the most promising solutions or substitutes for conventional fertilizers. These engineered materials are composed of nanoparticles containing macro- and micronutrients that are delivered to the plant rhizosphere in a regulated manner. In nanofertilizers, the essential minerals and nutrients (such as N, P, K, Fe, and Mn) are bonded alone or in combination with nano-dimensional adsorbents. This review discusses the development of nanotechnology-based smart and efficient agriculture using nanofertilizers that have higher nutritional management, owing to their ability to increase the nutrient uptake efficiency. Additionally, the synthesis and mechanism of action of the nanofertilizers are discussed, along with the different types of fertilizers that are currently available. Furthermore, sustainable agriculture can be realised by the targeted delivery and controlled release of nutrients through the application of nanoscale active substances. This paper emphasises the successful development and safe application of nanotechnology in agriculture; however, certain basic concerns and existing gaps in research need to be addressed and resolved.

Green nanotechnology is currently a very crucial and indispensable technology for handling diverse problems regarding the living planet. The concoction of reactive oxygen species (ROS) and biologically synthesized silver nanoparticles (AgNPs) has opened new insights in cancer therapy. The current investigation caters to the concept of the involvement of a novel eco-friendly avenue to produce AgNPs employing the wild endolichenic fungus Talaromyces funiculosus. The synthesized Talaromyces funiculosus -AgNPs were evaluated with the aid of UV visible spectroscopy, dynamic light scattering (DLS), Fourier infrared spectroscopy (ATR-FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The synthesized Talaromyces funiculosus -AgNPs ( TF -AgNPs) exhibited hemo-compatibility as evidenced by a hemolytic assay. Further, they were evaluated for their efficacy against foodborne pathogens Staphylococcus aureus, Streptococcus faecalis , Listeria innocua , and Micrococcus luteus and nosocomial Pseudomonas aeruginosa, Escherichia coli, Vibrio cholerae, and Bacillus subtilis bacterial strains. The synthesized TF -AgNPs displayed cytotoxicity in a dose-dependent manner against MDA-MB-231 breast carcinoma cells and eventually condensed the chromatin material observed through the Hoechst 33342 stain. Subsequent analysis using flow cytometry and fluorescence microscopy provided the inference of a possible role of intracellular ROS (OH - , O - , H 2 O 2 , and O 2 - ) radicals in the destruction of mitochondria, DNA machinery, the nucleus, and overall damage of the cellular machinery of breast cancerous cells. The combined effect of predation by the cyclopoid copepod Mesocyclops aspericornis and TF-AgNPS for the larval management of dengue vectors were provided. A promising larval control was evident after the conjunction of both predatory organisms and bio-fabricated nanoparticles. Thus, this study provides a novel, cost-effective, extracellular approach of TF -AgNPs production with hemo-compatible, antioxidant, and antimicrobial efficacy against both human and foodborne pathogens with cytotoxicity (dose dependent) towards MDA-MB-231 breast carcinoma.

Nongenomic effects of estrogen receptor α (ERα) signaling have been described for decades. Several distinct animal models have been generated previously to analyze the nongenomic ERα signaling (eg, membrane-only ER, and ERαC451A). However, the mechanisms and physiological processes resulting solely from nongenomic signaling are still poorly understood. Herein, we describe a novel mouse model for analyzing nongenomic ERα actions named H2NES knock-in (KI). H2NES ERα possesses a nuclear export signal (NES) in the hinge region of ERα protein resulting in exclusive cytoplasmic localization that involves only the nongenomic action but not nuclear genomic actions. We generated H2NESKI mice by homologous recombination method and have characterized the phenotypes. H2NESKI homozygote mice possess almost identical phenotypes with ERα null mice except for the vascular activity on reendothelialization. We conclude that ERα-mediated nongenomic estrogenic signaling alone is insufficient to control most estrogen-mediated endocrine physiological responses; however, there could be some physiological responses that are nongenomic action dominant. H2NESKI mice have been deposited in the repository at Jax (stock no. 032176). These mice should be useful for analyzing nongenomic estrogenic responses and could expand analysis along with other ERα mutant mice lacking membrane-bound ERα. We expect the H2NESKI mouse model to aid our understanding of ERα-mediated nongenomic physiological responses and serve as an in vivo model for evaluating the nongenomic action of various estrogenic agents., (Published by Oxford University Press on behalf of the Endocrine Society 2022.)

Pharmaceutical grade 3'-deoxy-3'-[ 18 F]fluorothymidine [ 18 F]FLT was synthesized using 3-N-Boc-5'-O-dimethoxytrityl-3'-O-nosyl-thymidine (BOC-Nosyl) precursor, in the general purpose TRACERlab FX modules. Purification of [ 18 F]FLT, via solid phase extraction (SPE) after radiosynthesis, using a combination of different SPE cartridges, yielded satisfactory results, with radiochemical and chemical purity >99%. While the non-decay corrected radiochemical yield (RCY) with 20 mg (24 μmole) of BOC-Nosyl precursor was found to be 6.80 ± 0.16%, the decay corrected radiochemical yield (RCY) was 9.95 ± 0.24%. Residual acetone, acetonitrile, and ethanol levels were found to be 22.97 ± 0.76, 109.08 ± 0.93, and 7,666.45 ± 3.7 ppm, respectively. A simplified method for solid-phase purification of [ 18 F]FLT was developed, circumventing the need for HPLC purification. Biodistribution in C57BL/6 mice with B16F10 cell line-induced melanoma showed tumor to blood ratio of ~3.8 at 90 min. PET/CT imaging of normal rabbit injected with [ 18 F]FLT shows selective uptake in the bone marrow and small intestine. [ 18 F]FLT was found to be excreted through the kidneys and get collected in the urinary bladder, 120 min post injection. PET/CT imaging performed in rabbit model at 30, 60, 90, and 120 min post [ 18 F]FLT injections showed concordance with tissue distribution kinetics of mice tumor model., (© 2022 John Wiley & Sons, Ltd.)

Exploring the molecular mechanisms behind bacterial adaptation to extreme temperatures has potential biotechnological applications. In the present study, Pseudomonas sp. Lz4W, a Gram-negative psychrophilic bacterium adapted to survive in Antarctica, was selected to decipher the molecular mechanism underlying the cold adaptation. Proteome analysis of the isolates grown at 4°C was performed to identify the proteins and pathways that are responsible for the adaptation. However, many proteins from the expressed proteome were found to be hypothetical proteins (HPs), whose function is unknown. Investigating the functional roles of these proteins may provide additional information in the biological understanding of the bacterial cold adaptation. Thus, our study aimed to assign functions to these HPs and understand their role at the molecular level. We used a structured insilico workflow combining different bioinformatics tools and databases for functional annotation. Pseudomonas sp. Lz4W genome (CP017432, version 1) contains 4493 genes and 4412 coding sequences (CDS), of which 743 CDS were annotated as HPs. Of these, from the proteome analysis, 61 HPs were found to be expressed consistently at the protein level. The amino acid sequences of these 61 HPs were submitted to our workflow and we could successfully assign a function to 18 HPs. Most of these proteins were predicted to be involved in biological mechanisms of cold adaptations such as peptidoglycan metabolism, cell wall organization, ATP hydrolysis, outer membrane fluidity, catalysis, and others. This study provided a better understanding of the functional significance of HPs in cold adaptation of Pseudomonas sp. Lz4W. Our approach emphasizes the importance of addressing the "hypothetical protein problem" for a thorough understanding of mechanisms at the cellular level, as well as, provided the assessment of integrating proteomics methods with various annotation and curation approaches to characterize hypothetical or uncharacterized protein data. The MS proteomics data generated from this study has been deposited to the ProteomeXchange through PRIDE with the dataset identifier-PXD029741., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ijaq, Chandra, Ray and Jagannadham.)

Exoribonuclease R (RNase R) is a 3' hydrolytic exoribonuclease that can degrade structured RNA. Mutation in RNase R affects virulence of certain human pathogenic bacteria. The aim of this study was to determine whether RNase R is necessary for virulence of the phytopathogen that causes bacterial blight in rice, Xanthomonas oryzae pv. oryzae (Xoo). In silico analysis has indicated that RNase R is highly conserved among various xanthomonads. Amino acid sequence alignment of Xoo RNase R with RNase R from various taxa indicated that Xoo RNase R clustered with RNase R of order Xanthomonadales. To study its role in virulence, we generated a gene disruption mutant of Xoo RNase R. The Xoo rnr - mutant is moderately virulence deficient, and the complementing strain ( rnr - /pHM1:: rnr ) rescued the virulence deficiency of the mutant. We investigated swimming and swarming motilities in both nutrient-deficient minimal media and nutrient-optimal media. We observed that RNase R mutation has adversely affected the swimming and swarming motilities of Xoo in optimal media. However, in nutrient-deficient media only swimming motility was noticeably affected. Growth curves in optimal media at suboptimal temperature (15°C cold stress) indicate that the Xoo rnr - mutant grows more slowly than the Xoo wild type and complementing strain ( rnr - /pHM1:: rnr ). Given these findings, we report for the first time that RNase R function is necessary for complete virulence of Xoo in rice. It is also important for motility of Xoo in media and for growth of Xoo at suboptimal temperature.

Objective: (1) Describe the progression of diabetes mellitus over time in an observational study of Wolfram syndrome, a rare, genetic, neurodegenerative disorder, which often includes diabetes mellitus and is typically diagnosed during childhood or adolescence. (2) Determine whether C-peptide could be used as a marker of diabetes progression in interventional trials for Wolfram syndrome., Methods: N = 44 (25F/19M) participants with genetically confirmed Wolfram syndrome attended the Washington University Wolfram Research Clinic annually from 2010 to 2019. Medical history, physical examinations, blood sampling, and questionnaires were used to collect data about diabetes mellitus and other components of Wolfram syndrome. Beta-cell function was assessed by determination of C-peptide during a mixed meal tolerance test. Random coefficients models evaluated the rate of progression of C-peptide over time, and power analyses were used to estimate the number of subjects needed to detect a change in C-peptide decline during an intervention trial., Results: 93.2% of patients had diabetes mellitus. Mean HbA1c across all study visits was 7.9%. C-peptide significantly decreased with increasing duration of diabetes mellitus (p < 0.0001); an optimal break point in C-peptide decline was identified to occur between 0.1 and 2.3 years after diabetes mellitus diagnosis. Twenty patients per group (active vs. control) were estimated to be needed to detect a 60% slowing of C-peptide decline during the first 2.3 years following diabetes diagnosis., Conclusion: C-peptide declines over time in Wolfram syndrome and could potentially be used as a marker of diabetes progression in interventional studies for Wolfram syndrome, especially within the first 2 years after diabetes diagnosis., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background and Aims: The study aimed to identify yet unknown and uncharacterized long non-coding RNAs (lncRNAs) in treatment-naïve ulcerative colitis (UC), and to define their possible roles in UC pathogenesis. For that purpose, accurate quantification methods for lncRNA transcript detection, multiple and "stringent" strategies were applied. New insights in the regulation of functional genes and pathways of relevance for UC through expression of lncRNAs are expected., Methods: The study was based on sequencing data derived from a data set consisting of treatment-naïve UC patients (n = 14) and control subjects (n = 16). Two complementary aligners were used to identify lncRNAs. Several different steps were used to validate differential expression including plotting the reads over the annotation for manual inspection. To help determine potential lncRNA involvement in biological processes, KEGG pathway enrichment was done on protein-coding genes which co-expressed with the lncRNAs., Results: A total of 99 lncRNAs were identified in UC. The lncRNAs which were not previously characterized (n = 15) in UC or other autoimmune diseases were selected for down-stream analysis. In total, 602 protein-coding genes correlated with the uncharacterized lncRNAs. KEGG pathway enrichment analysis revealed involvement of lncRNAs in two significantly enriched pathways, lipid and atherosclerosis, and T-cell receptor signaling., Conclusion: This study identified a set of 15 yet uncharacterized lncRNAs which may be of importance for UC pathogenesis. These lncRNAs may serve as potential diagnostic biomarkers and might be of use for the development of UC treatment strategies in the future., Competing Interests: The authors declare no conflict of interests regarding the publication of this manuscript., (© 2022 The Authors.)

Background/objective: Coronavirus disease 2019 (COVID-19) is a new disease; its clinical profile and natural history are evolving. Each well-recorded case in homeopathic practice is important for deciding the future course of action. This study aims at identifying clinically useful homeopathic remedies and their prescribing symptoms using the prognostic factor research model., Methods: This was an open-label, multi-centric, observational study performed from April 2020 to July 2020 at various public health care clinics. The data were collected prospectively from clinical practice at integrated COVID-19 care facilities in India. Good-quality cases were selected using a specific set of criteria. These cases were analyzed for elucidating prognostic factors by calculating the likelihood ratio (LR) of each frequently occurring symptom. The symptoms with high LR values (>1) were considered as prescribing indications of the specific remedy., Results: Out of 327 COVID-19 cases reported, 211 met the selection criteria for analysis. The most common complaints were fatigue, sore throat, dry cough, myalgia, fever, dry mouth and throat, increased thirst, headache, decreased appetite, anxiety, and altered taste. Twenty-seven remedies were prescribed and four of them- Arsenicum album, Bryonia alba, Gelsemium sempervirens , and Pulsatilla nigricans -were the most frequently used. A high LR was obtained for certain symptoms, which enabled differentiation between the remedies for a given patient., Conclusion: Homeopathic medicines were associated with improvement in symptoms of COVID-19 cases. Characteristic symptoms of four frequently indicated remedies have been identified using prognostic factor research, findings that can contribute to accurate homeopathic prescribing during future controlled research in COVID-19., Competing Interests: None declared., (Faculty of Homeopathy. This article is published by Thieme.)

Type 1 diabetes (T1D) is one of the most common chronic childhood diseases, and its prevalence is rapidly increasing. The management of glucose in T1D is challenging, as youth must consider a myriad of factors when making diabetes care decisions. This task often leads to significant hyperglycemia, hypoglycemia, and glucose variability throughout the day, which have been associated with short- and long-term medical complications. At present, most of what is known about each of these complications and the health behaviors that may lead to them have been uncovered in the clinical setting or in laboratory-based research. However, the tools often used in these settings are limited in their ability to capture the dynamic behaviors, feelings, and physiological changes associated with T1D that fluctuate from moment to moment throughout the day. A better understanding of T1D in daily life could potentially aid in the development of interventions to improve diabetes care and mitigate the negative medical consequences associated with it. Therefore, there is a need to measure repeated, real-time, and real-world features of this disease in youth. This approach is known as ecological momentary assessment (EMA), and it has considerable advantages to in-lab research. Thus, this viewpoint aims to describe EMA tools that have been used to collect data in the daily lives of youth with T1D and discuss studies that explored the nuances of T1D in daily life using these methods. This viewpoint focuses on the following EMA methods: continuous glucose monitoring, actigraphy, ambulatory blood pressure monitoring, personal digital assistants, smartphones, and phone-based systems. The viewpoint also discusses the benefits of using EMA methods to collect important data that might not otherwise be collected in the laboratory and the limitations of each tool, future directions of the field, and possible clinical implications for their use., (©Mary Katherine Ray, Alana McMichael, Maria Rivera-Santana, Jacob Noel, Tamara Hershey. Originally published in JMIR Diabetes (https://diabetes.jmir.org), 03.06.2021.)

Introduction: Risk assessment tools are routinely used to identify patients at high risk. There is increasing evidence that these tools may not be sufficiently accurate to determine the risk of suicide of people, particularly those being treated in community mental health settings., Methods: An outcome analysis for case serials of people who died by suicide between January 2014 and December 2016 and had contact with a public mental health service within 31 days prior to their death., Results: Of the 68 people who had contact, 70.5% had a formal risk assessment. Seventy-five per cent were classified as low risk of suicide. None were identified as being at high risk. While individual risk factors were identified, these did not allow to differentiate between patients classified as low or medium., Discussion: Risk categorisation contributes little to patient safety. Given the dynamic nature of suicide risk, a risk assessment should focus on modifiable risk factors and safety planning rather than risk prediction., Conclusion: The prediction value of suicide risk assessment tools is limited. The risk classifications of high, medium or low could become the basis of denying necessary treatment to many and delivering unnecessary treatment to some and should not be used for care allocation.

Background: Polyphenols have been studied for their potential involvement in the prevention of various chronic diseases as well as for their antimicrobial potential. The crude extracts of arecanut have been reported to have antiinfective properties. We aimed to explore the endosperm of Areca catechu (arecanut) for the extraction of polyphenol components and to study the antituberculosis activity of these polyphenol against Mycobacterium tuberculosis H37Rv., Method: A comparative extraction was performed using microwave and Soxlet apparatus. High performance liquid chromatography (HPLC) technique was used for the estimation of the extracted polyphenols. The minimum inhibitory concentration (MIC) values against M.tuberculosis H37Rv stain, Staphylococcus aureus and Escherichia coli were estimated by resazurin microtiter assay., Results: There was a 11-fold increase in the total phenolic content by microwave assisted extraction compared to the Soxhlet extraction. The powdered extract was found to be active with MIC value of 0.975 ± 0.02 μg/mL. Fractionation and HPLC-based estimation of the extract revealed catechin, epicatechin, and epigallocatechin gallate to be the polyphenol components in the ethanol fraction., Conclusions: The bioactivity of these polyphenols confirmed their presence and complementary effect in the extract form. Because the toxic alkaloid arecoline, known to be present in arecanut, did not show any activity individually, the bioactivity of the extract was attributed to the nontoxic polyphenols present. This extract also showed selective inhibition of M. tuberculosis over other gram positive and gram-negative bacteria, thereby establishing that arecanut is an exploitable selective source of polyphenols acting against M. tuberculosis., Competing Interests: None

Background: This study aimed to further elucidate correlated weight changes in parent-child dyads enrolled in family-based treatment (FBT) by modeling the interdependence of weight changes during treatment., Methods: Parent-child dyads (n = 172) with overweight/obesity (child mean zBMI = 2.16 ± 0.39; parent mean BMI = 37.9 ± 9.4 kg/m 2 ) completed 4 months of FBT and were randomized to one of three 8-month maintenance interventions (Social Facilitation Maintenance [SFM]-high dose, SFM-low dose or control). Weight/height was measured at 0, 4 and 12 months. Structural equation models simultaneously estimated the effect that an individual had on their own (actor effect) and on one another's (partner effect) weight-status across time using the actor-partner interdependence model., Results: Actor paths were significant over time for parent and child. Partner paths were significant for child zBMI predicting parent BMI at 4 and 12 months. Maintenance condition moderated actor/partner paths in the model., Conclusions: Child weight change may motivate parents to make environmental and behavioural changes that impact their own weight., Trial Registration: ClinicalTrials.gov Identifier: NCT00759746., (© 2020 World Obesity Federation.)

Purpose: To synthesize the literature in relation to findings of system errors through reviews of suicide deaths in the public mental health system., Data Sources: A systematic narrative meta-synthesis using the PRISMA methodology was conducted., Study Selection: All English language articles published between 2000 and 2017 that reported on system errors identified through reviews of suicide deaths were included. Articles that reported on patient factors, contact with General Practitioners or individual cases were excluded., Data Extraction: Results were extracted and summarized. An overarching coding framework was developed inductively. This coding framework was reapplied to the full data set., Results of Data Synthesis: Fourteen peer reviewed publications were identified. Nine focussed on suicide deaths that occurred in hospital or psychiatric inpatient units. Five studies focussed on suicide deaths while being treated in the community. Vulnerabilities were identified throughout the patient's journey (i.e. point of entry, transitioning between teams, and point of exit with the service) and centred on information gathering (i.e. inadequate and incomplete risk assessments or lack of family involvement) and information flow (i.e. transitions between different teams). Beyond enhancing policy, guidelines, documentation and regular training for frontline staff there were very limited suggestions as to how systems can make it easier for staff to support their patients., Conclusions: There are currently limited studies that have investigated learnings and recommendations. Identifying critical vulnerabilities in systems and to be proactive about these could be one way to develop a highly reliable mental health care system., (© The Author(s) 2020. Published by Oxford University Press in association with the International Society for Quality in Health Care. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Introduction: Social support for healthy eating can influence child eating behaviors; however, little is known about the impact of social support during family-based behavioral weight-loss treatment (FBT). This study aimed to determine the impacts of both baseline and change in family support on change in child diet and weight during FBT. Methods: Children ( n  = 175; BMI percentile ≥85th; ages 7-11; 61.1% female; 70.9% white) and a participating parent completed 4 months of FBT. Parents were active participants and learned social support-related strategies ( i.e. , praise and modeling of healthy eating). Child perceived family encouragement and discouragement for healthy eating, child diet quality (via 24-hour recalls), and child weight were assessed pre- and post-FBT. Results: Family encouragement for healthy eating increased during FBT, and this increase was related to increases in child healthy vegetable intake and overall diet quality, as well as decreases in refined grains consumed. Low pre-FBT family encouragement predicted greater increases in healthy vegetable intake, greater weight reduction, and greater increases in family encouragement for healthy eating. Family discouragement for healthy eating did not change during treatment nor did it predict dietary or weight outcomes. Conclusions: FBT successfully improves family encouragement, which is associated with improvements in child diet. Furthermore, even children who began treatment with low family encouragement for healthy eating show great improvements in dietary intake and weight during treatment. Results suggest that changes in child eating behavior during treatment is influenced by active, positive parenting techniques such as praise of healthy eating rather than negative family support.

Differences in trait suggestibility among those with obesity may help explain differential responses to weight-loss interventions. Ubiquitous advertising of unhealthy foods, weight-loss products that are not evidence-based, and myths regarding weight loss could be particularly sabotaging in individuals with high levels of suggestibility, with or at risk of developing obesity. This study explored relationships between suggestibility, body mass index (BMI), and self-reported eating-related behaviors that vary among those with obesity. A sample of ethnically diverse adults (N = 73) with a BMI ≥25 completed the Short Suggestibility Scale (SSS), Palatable Eating Motives Scale (PEMS), Binge Eating Scale (BES), Dutch Eating Behavior Questionnaire-Restraint (DEBQ-R), and Barratt Impulsiveness Scale (BIS). Impulsiveness was controlled in analyses due to its strong association with suggestibility. Analyses revealed that BMI was not related to SSS scores, consistent with studies using hypnotic-suggestibility scales. However, SSS scores were positively associated with eating caloric food more frequently for Reward, Social, and Conformity motives, and with greater actual dieting behavior, and binge eating. Suggestibility was not related to eating for Coping motives or effort to diet. If supported by future replications, knowledge of these associations could potentially help inform and tailor weight-loss interventions to protect those that may be most susceptible to adopting invalid messages and products., Competing Interests: Declaration of competing interest All the authors declared that they have no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: The ability of health professions faculty to design, teach, evaluate, and improve relevant curricula is vital for teaching improvement science (IS) skills to trainees., Objective: We launched a Foundational Improvement Science Curriculum (FISC) to build faculty competence in IS teaching and scholarship, and to develop, expand, and standardize IS curricula across one institution., Methods: FISC consisted of 9 full or half-day sessions over 10 months in 2015-2016 and 2016-2017 academic years. Each session required pre-work, including readings, Institute for Healthcare Improvement Open School modules, and personal improvement projects. Sessions included brief didactics, group activities, planning, and feedback on curriculum development. An evaluation strategy was employed, including pre- and post-program self-assessment, competency mapping, evaluations of didactics and overall program, and participant satisfaction., Results: Forty individuals from 23 academic programs voluntarily completed FISC, representing 20% of graduate medical education (GME) programs and 50% of primary GME programs in addition to undergraduate medical education (UME) and nursing programs. Median self-assessed competency scores (mid versus final score; scale 1-9, 9 high; P < .05 for all comparisons) improved over the course for all competencies for knowledge (3 versus 7), application (2 versus 7), curriculum design (2 versus 7), and scholarship (2 versus 5). Eighteen new or revised IS curricula were developed across GME, UME, and nursing programs., Conclusions: FISC offers a feasible model to enhance and support faculty development in IS and IS curriculum design., Competing Interests: Conflict of interest: The authors declare they have no competing interests., (Accreditation Council for Graduate Medical Education 2019.)

5-Substituted-6-acetyl-2-amino-7-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-4(3H)-one derivatives were synthesized and evaluated against Mycobacterium tuberculosis H37Rv, Mycobacterium aurum, Escherichia coli, and Staphylococcus aureus as well as a human monocyte-derived macrophage (THP-1), and murine macrophage (RAW 264.7) cell lines to assess their antibacterial and cytotoxic potential, respectively. The compounds showed activity in the range of 1.95-125 µg/ml against M. tuberculosis but showed no activity against M. aurum, E. coli, and S. aureus, indicating selectivity towards slow-growing mycobacterial pathogens. The compounds exhibited very low to no cytotoxicity up to 500 µg/ml concentration against eukaryotic cell lines. The most potent molecule, 2l, showed a minimum inhibitory concentration of 1.95 µg/ml against M. tuberculosis H37Rv and a selectivity index of >250 against both the eukaryotic cell lines. Furthermore, 2l showed moderate inhibition of whole-cell mycobacterial drug-efflux pumps when compared to verapamil, a known potent inhibitor of efflux pumps. Thus, derivative 2l was identified as an antituberculosis hit molecule, which could be used to yield more potent lead molecules., (© 2019 Deutsche Pharmazeutische Gesellschaft.)

Purpose: Behavioral predictors of weight-loss program (WLP) outcomes are needed and important because they can be modified. Eating calorie-dense palatable foods (PFs) outside of hunger contributes to obesity. This study assessed if habitual motives to consume PFs could predict weight-loss outcomes., Methods: N = 171 Black and N = 141 White adults in a reduced-calorie program completed the Palatable Eating Motives Scale (PEMS). Body weight and body mass index (BMI) lost after 3 and 6 months were analyzed controlling for initial BMI and demographics. Greater PEMS motive scores meant more frequent habitual intake of PFs for that motive., Results: Whites vs. Blacks had higher scores on most of the PEMS motives: Social, Coping, and Reward Enhancement. In Whites at 3 months, greater Reward Enhancement scores and initial BMI predicted more BMI loss (p < 0.05). At 6 months, greater Reward Enhancement and lower Conformity scores predicted more weight (p < 0.05) and BMI loss (Conformity: p < 0.05; Reward Enhancement: p = 0.05). PEMS motives did not predict outcomes for Blacks., Conclusion: The results provide preliminary evidence for the PEMS to predict WLP outcomes. White patients who eat PFs primarily for their rewarding properties and less to conform should fare better in Lifestyle programs while group or family-based interventions may be more efficacious when conformity is the main motive. Lower motive scores among Blacks suggest that eating PFs outside of hunger may go unrecognized or underreported and warrants further investigation. The findings highlight the motive-based heterogeneity of obesity and how it may be used to predict outcomes and customize interventions to improve WLP outcomes., Level of Evidence: Level IV, multiple time series.

Transcranial direct current stimulation (tDCS) is a neuromodulation technique with potential to treat eating disorders and obesity. As for any potential treatment, it is important to assess the degree to which expectation effects contribute to its reported efficacy. This study assessed the effect of tDCS on amount of food craving and eating while tightly controlling treatment expectation. N = 74 adults with overweight or obesity were informed of the known effects of tDCS to suppress craving and eating. Once electrodes were on the head, half of the participants were told they were receiving real, and the other half sham tDCS. Within these groups, approximately half actually received real and the other half sham tDCS. Stimulation parameters used were those previously found to reduce craving and eating, including in our lab: 2 mA, anode right/cathode left targeting the dorsolateral prefrontal cortex for 20 min (real), or only for the first and last minute (sham). Analyses controlled for demographics, hunger, trait impulsiveness, eating motives, dieting, binge eating, suggestibility, and baseline craving and eating. Participants told they were receiving real tDCS craved and ate less than participants told they were receiving sham tDCS (both p < 0.01), regardless of tDCS condition administered. There was no main effect of real vs. sham tDCS on craving or eating or an interaction between tDCS condition and expectation. The scientific validation of tDCS as a treatment for eating-related conditions hinges on controlling for the powerful effects of expectation. This can include the type of information provided on consent forms and participants' ability to guess real from sham conditions., (Published by Elsevier Ltd.)

The study of gene function in normal human physiology and pathophysiology is complicated by countless factors such as genetic diversity (~98 million SNPs identified in the human genome as of June 2015), environmental exposure, epigenetic imprinting, maternal/in utero exposure, diet, exercise, age, sex, socioeconomic factors, and many other variables. Inbred mouse lines have allowed researchers to control for many of the variables that define human diversity but complicate the study of the human genome, gene/protein function, cellular and molecular pathways, and countless other genetic diseases. Furthermore, genetically modified mouse models enable us to generate and study mice whose genomes differ by as little as a single point mutation while controlling for non-genomic variables. This allows researchers to elucidate the quintessential function of a gene, which will further not only our scientific understanding, but provide key insight into human health and disease. Recent advances in CRISPR/Cas9 genome editing have revolutionized scientific protocols for introducing mutations into the mammalian genome. The ensuing chapter provides an overview of CRISPR/Cas9 genome editing in murine embryonic stem cells for the generation of genetically modified mouse models.

The identity of the gonads is determined by which fate, ovarian granulosa cell or testicular Sertoli cell, the bipotential somatic cell precursors choose to follow. In most vertebrates, the conserved transcription factor FOXL2 contributes to the fate of granulosa cells. To understand FOXL2 functions during gonad differentiation, we performed genome-wide analysis of FOXL2 chromatin occupancy in fetal ovaries and established a genetic mouse model that forces Foxl2 expression in the fetal testis. When FOXL2 was ectopically expressed in the somatic cell precursors in the fetal testis, FOXL2 was sufficient to repress Sertoli cell differentiation, ultimately resulting in partial testis-to-ovary sex-reversal. Combining genome-wide analysis of FOXL2 binding in the fetal ovary with transcriptomic analyses of our Foxl2 gain-of-function and previously published Foxl2 loss-of-function models, we identified potential pathways responsible for the feminizing action of FOXL2. Finally, comparison of FOXL2 genome-wide occupancy in the fetal ovary with testis-determining factor SOX9 genome-wide occupancy in the fetal testis revealed extensive overlaps, implying that antagonistic signals between FOXL2 and SOX9 occur at the chromatin level.