An optimized CT-dense agent perfusion and micro-CT imaging protocol for chick embryo developmental stages.

Compared to classical techniques of morphological analysis, micro-CT (µ-CT) has become an effective approach allowing rapid screening of morphological changes. In the present work, we aimed to provide an optimized micro-CT dense agent perfusion protocol and µ-CT guidelines for different stages of chick embryo cardiogenesis. Our study was conducted over a period of 10 embryonic days (Hamburger-Hamilton HH36) in chick embryo hearts. During the perfusion of the micro-CT dense agent at different developmental stages (HH19, HH24, HH27, HH29, HH31, HH34, HH35, and HH36), we demonstrated that durations and volumes of the injected contrast agent gradually increased with the heart developmental stages contrary to the flow rate that was unchanged during the whole experiment. Analysis of the CT imaging confirmed the efficiency of the optimized parameters of the heart perfusion.

Interband cascade lasers grown simultaneously on GaSb, GaAs and Si substrates.

We report on Sb-based interband cascade lasers simultaneously grown on GaSb, GaAs and Si substrates. 8 µm x 2 mm devices exhibited similar threshold currents around 40 mA at 20°C and achieved continuous-wave (CW) operation up to 65°C on GaSb, GaAs and Si substrates despite a dislocation density of ∼ 4.108 cm-2 for both mismatched substrates. In the CW regime the output power of the devices emitting at 3.3 µm exceeded 30 mW/facet at 20°C. ICLs on GaAs and Si were subsequently aged at 50°C with an injection current of 200 mA, i.e. five times the laser-threshold current. No degradation was observed after 500 h of CW operation, demonstrating the high performance of ICLs and their tolerance to dislocations.

Validation of a modified QuEChERS method for the quantification of residues of currently used pesticides in Cuban agricultural soils, using gas chromatography tandem mass spectrometry.

We present an analytical method to detect and quantify residues of currently used pesticides (CUPs), which include 31 active ingredients (ai) and seven transformation products (TPs) in tropical and agricultural soils of Cuba. Ten isotopically labeled analogous compounds served as internal standards (IL-IS). The novelty of this research is the inclusion of different tropical soils type scarcely studied for CUPs and TPs, based on the QuEChERS (quick, easy, cheap, effective, rugged and safe) method, followed by chromatography tandem mass spectrometry. All figures of merit proved to be satisfactory according to SANTE guidelines 2020 and 2021. Matrix effects (ME) calculated by the external standard method were significant (|ME| > 20% for almost all compounds; grand mean ± standard deviation (STD) 104 ± 108%) in all soils. The internal standard method compensated ME to non-significant levels (8 ± 50%), even for analytes with a non-structure identical IL-IS (STD, 13 ± 57%). Repeatability (relative standard deviation, RSDr) and reproducibility (RSDR) for skeletic regosol (SR) were 7.5 ± 2.8% and 11.7 ± 4.7%, respectively. Absolute (quantified for 11 analytes with structure identical IL-IS) and relative recovery from SR was 92 ± 13% (mean ± STD) and 90 ± 12%, respectively. Limits of quantification for SR ranged from 0.1 to 10 ng/g, except metalaxyl and oxyfluorfen (25 ng/g each). Linearity of matrix-matched (MM) calibration curves (5 to 100 ng/g) had an R2 of ≥ 0.99 for all soils and almost all analytes. The method was successfully applied to 30 real soil samples.

Different Nuclear Architecture in Human Sperm According to Their Morphology.

Human sperm parameters serve as a first step in diagnosing male infertility, but not in determining the potential for successful pregnancy during assisted reproductive technologies (ARTs) procedures. Here, we investigated the relationship between sperm head morphology at high magnification, based on strict morphologic criteria, and the nuclear architecture analyzed by fluorescence in situ hybridization (FISH). We included five men. Two of them had an elevated high-magnification morphology score of 6 points (Score 6) indicating high fertility potential, whereas three had a low score of 0 points (Score 0), indicating low fertility potential. We used FISH to study the inter-telomeric distance and the chromosomal territory area of chromosome 1 (Chr. 1). We then compared these two parameters between subjects with high and low scores. FISH data analysis showed that the inter-telomeric distance (ITD) and chromosomal territory area (CTA) of Chr. 1 were significantly higher in subjects with low scores (score 0) than high scores (score 6). Our results suggest that (i) there is a link between nuclear architecture and sperm head abnormalities, particularly vacuoles; and (ii) it is possible to select spermatozoa with normal nuclear architecture, which might indirectly explain the positive ART outcomes observed with this technique.

Metabolic reprogramming of the retinal pigment epithelium by cytokines associated with age-related macular degeneration.

The complex metabolic relationship between the retinal pigment epithelium (RPE) and photoreceptors is essential for maintaining retinal health. Recent evidence indicates the RPE acts as an adjacent lactate sink, suppressing glycolysis in the epithelium in order to maximize glycolysis in the photoreceptors. Dysregulated metabolism within the RPE has been implicated in the pathogenesis of age-related macular degeneration (AMD), a leading cause of vision loss. In the present study, we investigate the effects of four cytokines associated with AMD, TNFα, TGF-ß2, IL-6, and IL-1ß, as well as a cocktail containing all four cytokines, on RPE metabolism using ARPE-19 cells, primary human RPE cells, and ex vivo rat eyecups. Strikingly, we found cytokine-specific changes in numerous metabolic markers including lactate production, glucose consumption, extracellular acidification rate, and oxygen consumption rate accompanied by increases in total mitochondrial volume and ATP production. Together, all four cytokines could potently override the constitutive suppression of glycolysis in the RPE, through a mechanism independent of PI3K/AKT, MEK/ERK, or NF-κB. Finally, we observed changes in glycolytic gene expression with cytokine treatment, including in lactate dehydrogenase subunit and glucose transporter expression. Our findings provide new insights into the metabolic changes in the RPE under inflammatory conditions and highlight potential therapeutic targets for AMD.

Paternal dietary macronutrient balance and energy intake drive metabolic and behavioral differences among offspring.

Paternal diet can influence the phenotype of the next generation, yet, the dietary components inducing specific responses in the offspring are not identified. Here, we use the Nutritional Geometry Framework to determine the effects of pre-conception paternal dietary macronutrient balance on offspring metabolic and behavioral traits in mice. Ten isocaloric diets varying in the relative proportion of protein, fats, and carbohydrates are fed to male mice prior to mating. Dams and offspring are fed standard chow and never exposed to treatment diets. Body fat in female offspring is positively associated with the paternal consumption of fat, while in male offspring, an anxiety-like phenotype is associated to paternal diets low in protein and high in carbohydrates. Our study uncovers that the nature and the magnitude of paternal effects are driven by interactions between macronutrient balance and energy intake and are not solely the result of over- or undernutrition.

IDH-mutant myeloid neoplasms are associated with seronegative rheumatoid arthritis and innate immune activation.

ABSTRACT: High prevalence of IDH mutations in seronegative rheumatoid arthritis (RA) with myeloid neoplasm, elevated 2-hydroxyglutarate, dysregulated innate immunity, and proinflammatory microenvironment suggests causative association between IDH mutations and seronegative RA. Our findings merit investigation of IDH inhibitors as therapeutics for seronegative IDH-mutated RA.

Developing a best-practice agenda for music therapy research to support informal carers of terminally ill patients pre- and post-death bereavement: a world café approach.

BACKGROUND: Informal carers of terminally ill patients play a vital role in providing palliative care at home, which impacts on their pre- and post-death bereavement experience and presents an up to 50% greater risk for mental-health problems. However, developing and implementing effective bereavement support remains challenging. There is a need to build the evidence base for music therapy as a potentially promising bereavement support for this vulnerable population. This study aimed to co-design an international best practice agenda for research into music therapy for informal carers of patients pre- and post-death bereavement. METHODS: Online half day workshop using a World Café approach; an innovative method for harnessing group intelligence within a group of international expert stakeholders (music therapy clinicians and academics with experience of music therapy with informal carers at end-of-life). Demographics, experience, key priorities and methodological challenges were gathered during a pre-workshop survey to inform workshop discussions. The online workshop involved four rounds of rotating, 25-minute, small group parallel discussions using Padlet. One final large group discussion involved a consensus building activity. All data were analysed thematically to identify patterns to inform priorities and recommendations. RESULTS: Twenty-two consented and completed the pre-event survey (response rate 44%), from countries representing 10 different time zones. Sixteen participated in the workshop and developed the following best practice agenda. The effectiveness of music therapy in supporting informal carers across the bereavement continuum should be prioritised. This should be done using a mixed methods design to draw on the strengths of different methodological approaches to building the evidence base. It should involve service users throughout and should use a core outcome set to guide the choice of clinically important bereavement outcome measures in efficacy/effectiveness research. CONCLUSIONS: Findings should inform future pre- and post-death bereavement support research for informal caregivers of terminally ill patients. This is an important step in building the evidence base for commissioners and service providers on how to incorporate more innovative approaches in palliative care bereavement services.

Tobacco smoking and diabetes. A comparative survey among diabetologists and smoking cessation specialists.

AIMS: Because tobacco smoking is a major risk factor of mortality in diabetes and guidelines suggest evaluating smoking behavior among individuals with diabetes and helping smokers quit, we aimed to assess knowledge about the tobacco smoking - diabetes relationship among diabetologists and smoking cessation specialists (SCS). METHODS: An online cross sectional survey was conceived by the Working Group on Smoking and Diabetes, France. The questionnaire was tested by the members of the Working Group and deemed to be completed in less than 5 min. Only questions receiving the highest number of approval ratings were kept for the survey. The questionnaire was sent to all members of the French Language Society of Diabetes (Société Francophone du Diabète, SFD), N = 969 and the French Language Society on Tobacco (Société Francophone de Tabacologie, SFT), N = 307. The mailing lists of members were obtained with the previous agreement of the societies' board. RESULTS: 225 diabetologists and 97 SCS (response rate 23.2% and 31.5%, respectively) completed the questionnaire. Over 90% of the diabetologists reported recording smoking status of their patients. Although diabetologists were aware that smoking increases all-cause mortality of individuals with diabetes, only 29.3% were aware that smoking is a risk factor for type 2 diabetes (76.3% among SCS), for poor glycemic control: 32.9% (86.6% among SCS). Significantly less diabetologists (64%) than SCS (76.3%) were aware of smoking being a risk factor for microangiopathy. More diabetologists considered that smoking cessation is more important than optimizing glycemic control among individuals with type 2 (69.3%) than among those with type 1 diabetes (47.1%). Few diabetologists (11.1%) and SCS (14.4%) reported to be trained for smoking cessation among persons with diabetes. CONCLUSION: Specific knowledge about the negative tobacco smoking - diabetes association seems to be insufficient among French diabetologists. Diabetologists but also other health care professionals should be trained to help individuals with diabetes who smoke to quit smoking.

Mitochondrial translocation of TFEB regulates complex I and inflammation.

TFEB is a master regulator of autophagy, lysosome biogenesis, mitochondrial metabolism, and immunity that works primarily through transcription controlled by cytosol-to-nuclear translocation. Emerging data indicate additional regulatory interactions at the surface of organelles such as lysosomes. Here we show that TFEB has a non-transcriptional role in mitochondria, regulating the electron transport chain complex I to down-modulate inflammation. Proteomics analysis reveals extensive TFEB co-immunoprecipitation with several mitochondrial proteins, whose interactions are disrupted upon infection with S. Typhimurium. High resolution confocal microscopy and biochemistry confirms TFEB localization in the mitochondrial matrix. TFEB translocation depends on a conserved N-terminal TOMM20-binding motif and is enhanced by mTOR inhibition. Within the mitochondria, TFEB and protease LONP1 antagonistically co-regulate complex I, reactive oxygen species and the inflammatory response. Consequently, during infection, lack of TFEB specifically in the mitochondria exacerbates the expression of pro-inflammatory cytokines, contributing to innate immune pathogenesis.

Is heated tobacco a harm reduction tool? / Le tabac chauffé est-il un outil de réduction des risques ?

Heated tobacco (HT), a new tobacco product, is presented by the tobacco industry as an effective and safe alternative to cigarettes. Even if the quantities of harmful compounds emitted by HT are lower than those found in cigarette smoke, this reduction in exposure cannot be equated with a reduction in risk. No study has provided evidence that switching from cigarettes to HT reduces the risk of tobacco-related diseases. HT cannot be considered as a cigarette cessation product and was even designed as a product to initiate or return to tobacco consumption. To promote this product, the tobacco industry essentially exploits the concept of harm reduction and, as such, tries in its commercial communication to create confusion between HT and electronic cigarettes, despite these two products having nothing in common. This promotion is based, on the one hand, on the data of internal studies in contradiction with those of independent studies, and, on the other, illegally, on social networks and communication in contradiction with the statements of regulation authorities. HT is a new lure offered by the tobacco industry, intended to maintain its profits in a world that is moving away from "traditional" cigarettes. It should be strictly advised against for both non-smokers and smokers.

Le tabac chauffé (TC), nouveau produit du tabac, est présenté par l'industrie du tabac comme une alternative efficace et sûre à la cigarette. Même si les quantités de composés nocifs pour la santé émises par le TC sont inférieures à celles trouvées dans la fumée de cigarette, cette diminution d'exposition ne peut être assimilée à une réduction des risques. Aucune étude n'a apporté la preuve que le passage de la cigarette au TC réduisait le risque des maladies liées au tabac. Le TC ne peut pas être considéré comme un produit de sevrage de la cigarette et a même été conçu comme un produit pour entrer dans la consommation de tabac ou y revenir. Pour faire la promotion de ce produit, l'industrie du tabac exploite essentiellement le concept de réduction des risques et, à ce titre, essaie dans sa communication commerciale de créer la confusion entre le TC et la cigarette électronique alors que ces deux produits n'ont rien à voir. Cette promotion s'appuie, d'une part, sur les données d'études internes contradictoires avec celles d'études indépendantes, d'autre part, de façon illégale, sur les réseaux sociaux et une communication en contradiction avec les avis des autorités de régulation. Le TC est un nouveau leurre de l'industrie du tabac, destiné à assurer le maintien de ses profits dans un monde qui s'éloigne de la cigarette « traditionnelle ¼. Il doit être formellement déconseillé, tant aux non-fumeurs qu'aux fumeurs.

IDH1-Mutant Preleukemic Hematopoietic Stem Cells Can Be Eliminated by Inhibition of Oxidative Phosphorylation.

Rare preleukemic hematopoietic stem cells (pHSC) harboring only the initiating mutations can be detected at the time of acute myeloid leukemia (AML) diagnosis. pHSCs are the origin of leukemia and a potential reservoir for relapse. Using primary human samples and gene editing to model isocitrate dehydrogenase 1 (IDH1) mutant pHSCs, we show epigenetic, transcriptional, and metabolic differences between pHSCs and healthy hematopoietic stem cells (HSC). We confirm that IDH1-driven clonal hematopoiesis is associated with cytopenia, suggesting an inherent defect to fully reconstitute hematopoiesis. Despite giving rise to multilineage engraftment, IDH1-mutant pHSCs exhibited reduced proliferation, blocked differentiation, downregulation of MHC class II genes, and reprogramming of oxidative phosphorylation metabolism. Critically, inhibition of oxidative phosphorylation resulted in the complete eradication of IDH1-mutant pHSCs but not IDH2-mutant pHSCs or wild-type HSCs. Our results indicate that IDH1-mutant preleukemic clones can be targeted with complex I inhibitors, offering a potential strategy to prevent the development and relapse of leukemia. SIGNIFICANCE: A high burden of pHSCs is associated with worse overall survival in AML. Using single-cell sequencing, metabolic assessment, and gene-edited human models, we find human pHSCs with IDH1 mutations to be metabolically vulnerable and sensitive to eradication by complex I inhibition. See related commentary by Steensma.

IDH1-mutant preleukemic hematopoietic stem cells can be eliminated by inhibition of oxidative phosphorylation.

Rare preleukemic hematopoietic stem cells (pHSCs) harboring only the initiating mutations can be detected at the time of AML diagnosis. pHSCs are the origin of leukemia and a potential reservoir for relapse. Using primary human samples and gene-editing to model isocitrate dehydrogenase 1 (IDH1) mutant pHSCs, we show epigenetic, transcriptional, and metabolic differences between pHSCs and healthy hematopoietic stem cells (HSCs). We confirm that IDH1 driven clonal hematopoiesis is associated with cytopenia, suggesting an inherent defect to fully reconstitute hematopoiesis. Despite giving rise to multilineage engraftment, IDH1-mutant pHSCs exhibited reduced proliferation, blocked differentiation, downregulation of MHC Class II genes, and reprogramming of oxidative phosphorylation metabolism. Critically, inhibition of oxidative phosphorylation resulted in complete eradication of IDH1-mutant pHSCs but not IDH2-mutant pHSCs or wildtype HSCs. Our results indicate that IDH1-mutant preleukemic clones can be targeted with complex I inhibitors, offering a potential strategy to prevent development and relapse of leukemia.

Application of a new multi-locus variable number tandem repeat analysis (MLVA) scheme for the seasonal investigation of Cryptosporidium parvum cases in Wales and the northwest of England, spring 2022.

The protozoan Cryptosporidium parvum is an important cause of gastroenteritis in humans and livestock, and cryptosporidiosis outbreaks are common. However, a multi-locus genotyping scheme is not widely adopted. We describe the further development and application of a seven-locus multi-locus variable number of tandem repeats analysis (MLVA) scheme. From 28th March to 31st July 2022, confirmed C. parvum stools (n = 213) from cryptosporidiosis patients (cases) in Wales (n = 95) and the north west of England (n = 118) were tested by MLVA. Typability (defined as alleles identified at all seven loci in a sample) was 81.2% and discriminatory power estimated by Hunter Gaston Discriminatory Index was 0.99. A MLVA profile was constructed from the alleles, expressed in chromosomal order. Profiles were defined as simple (single allele at each locus) or mixed (more than one allele at any locus). A total of 161 MLVA profiles were identified; 13 were mixed, an additional 38 simple profiles contained null records, and 110 were complete simple profiles. A minimum spanning tree was constructed of simple MLVA profiles and those identical at all seven loci defined genetic clusters of cases (here, null records were considered as an allele); 77 cases formed 25 clusters, ranging from two to nine (mode = two) cases. The largest cluster, following epidemiological investigation, signalled a newly-identified outbreak. Two other cases with mixed profiles that contained the outbreak alleles were included in the outbreak investigation. In another epidemiologically-identified outbreak of six initial cases, MLVA detected two additional cases. In a third, small outbreak of three cases, identical MLVA profiles strengthened the microbiological evidence. Review of the performance characteristics of the individual loci and of the seven-locus scheme suggested that two loci might be candidates for review, but a larger dataset over a wider geographical area and longer timeframe will help inform decision-making about the scheme by user laboratories and stakeholders (such as public health agencies). This MLVA scheme is straightforward in use, fast and cheap compared to sequence-based methods, identifies mixed infections, provides an important tool for C. parvum surveillance, and can enhance outbreak investigations and public health action.

An outbreak of Salmonella Typhimurium associated with the consumption of raw liver at an Eid al-Adha celebration in Wales (UK), July 2021.

In July 2021, Public Health Wales received two notifications of salmonella gastroenteritis. Both cases has attended the same barbecue to celebrate Eid al-Adha, two days earlier. Additional cases attending the same barbecue were found and an outbreak investigation was initiated. The barbecue was attended by a North African community's social network. On same day, smaller lunches were held in three homes in the social network. Many people attended both a lunch and the barbecue. Cases were defined as someone with an epidemiological link to the barbecue and/or lunches with diarrhoea and/or vomiting with date of onset following these events. We undertook a cohort study of 36 people attending the barbecue and/or lunch, and a nested case-control study using Firth logistic regression. A communication campaign, sensitive towards different cultural practices, was developed in collaboration with the affected community. Consumption of a traditional raw liver dish, 'marrara', at the barbecue was the likely vehicle for infection (Firth logistic regression, aOR: 49.99, 95%CI 1.71-1461.54, p = 0.02). Meat and offal came from two local butchers (same supplier) and samples yielded identical whole genome sequences as cases. Future outbreak investigations should be relevant to the community affected by considering dishes beyond those found in routine questionnaires.

Covid and the coalfield: Covid-19 vaccine hesitance in Wales and Appalachia.

BACKGROUND: Vaccine hesitancy is a barrier to Covid-19 vaccine uptake and displays a social gradient, compounding health disparities. While social gradients are a vital concept in health, they flatten distinctions between types of disadvantaged community. This paper focuses on vaccine hesitance in post-industrial and de-industrialising coalfields. The social consequences of the decline of coal mining may present barriers to vaccine uptake. METHODS: We ran parallel surveys in Wales (N = 4187) and US states overlapping with central Appalachia (N = 4864), to examine whether vaccine attitudes and uptake varied between areas with different coal mining histories. These surveys were accompanied by qualitative interviews of 36 residents of these coalfields to explore vaccination decisions and triangulate with survey data. RESULTS: Factor analysis identified four axes of attitudes in the survey data: vaccine confidence, covid scepticism, vaccine individualism, and concerned confusion. These themes were echoed in the interviews. Vaccine confidence was lower; and covid scepticism, vaccine individualism, and concerned confusion higher, in residents of areas of Wales with greater mining extent and where pits closed during certain periods. Residents of former US coal counties had lower vaccine confidence and higher covid scepticism, while those in current coal counties had greater vaccine individualism and concerned confusion. In former US coal counties and Welsh areas where pits closed since 1980, vaccine uptake was lower. Differences could not be explained by respondents' income and education. In the interviews, norms of social solidarity were often invoked by vaccinated respondents, while unvaccinated respondents did not frame decisions in the context of the industrial history of their areas. DISCUSSION: The legacy of coal-mining's decline presents barriers to public health campaigns. We show evidence of this across two historically significant coalfields. Attention is needed to avert negative public health consequences of global energy transition.