A concerted neuron-astrocyte program declines in ageing and schizophrenia.

Human brains vary across people and over time; such variation is not yet understood in cellular terms. Here we describe a relationship between people's cortical neurons and cortical astrocytes. We used single-nucleus RNA sequencing to analyse the prefrontal cortex of 191 human donors aged 22-97 years, including healthy individuals and people with schizophrenia. Latent-factor analysis of these data revealed that, in people whose cortical neurons more strongly expressed genes encoding synaptic components, cortical astrocytes more strongly expressed distinct genes with synaptic functions and genes for synthesizing cholesterol, an astrocyte-supplied component of synaptic membranes. We call this relationship the synaptic neuron and astrocyte program (SNAP). In schizophrenia and ageing-two conditions that involve declines in cognitive flexibility and plasticity1,2-cells divested from SNAP: astrocytes, glutamatergic (excitatory) neurons and GABAergic (inhibitory) neurons all showed reduced SNAP expression to corresponding degrees. The distinct astrocytic and neuronal components of SNAP both involved genes in which genetic risk factors for schizophrenia were strongly concentrated. SNAP, which varies quantitatively even among healthy people of similar age, may underlie many aspects of normal human interindividual differences and may be an important point of convergence for multiple kinds of pathophysiology.

Repeated sensitization of mice with microfilariae of Litomosoides sigmodontis induces pulmonary eosinophilia in an IL-33-dependent manner.

BACKGROUND: Eosinophilia is a hallmark of helminth infections and eosinophils are essential in the protective immune responses against helminths. Nevertheless, the distinct role of eosinophils during parasitic filarial infection, allergy and autoimmune disease-driven pathology is still not sufficiently understood. In this study, we established a mouse model for microfilariae-induced eosinophilic lung disease (ELD), a manifestation caused by eosinophil hyper-responsiveness within the lung. METHODS: Wild-type (WT) BALB/c mice were sensitized with dead microfilariae (MF) of the rodent filarial nematode Litomosoides sigmodontis three times at weekly intervals and subsequently challenged with viable MF to induce ELD. The resulting immune response was compared to non-sensitized WT mice as well as sensitized eosinophil-deficient dblGATA mice using flow cytometry, lung histology and ELISA. Additionally, the impact of IL-33 signaling on ELD development was investigated using the IL-33 antagonist HpARI2. RESULTS: ELD-induced WT mice displayed an increased type 2 immune response in the lung with increased frequencies of eosinophils, alternatively activated macrophages and group 2 innate lymphoid cells, as well as higher peripheral blood IgE, IL-5 and IL-33 levels in comparison to mice challenged only with viable MF or PBS. ELD mice had an increased MF retention in lung tissue, which was in line with an enhanced MF clearance from peripheral blood. Using eosinophil-deficient dblGATA mice, we demonstrate that eosinophils are essentially involved in driving the type 2 immune response and retention of MF in the lung of ELD mice. Furthermore, we demonstrate that IL-33 drives eosinophil activation in vitro and inhibition of IL-33 signaling during ELD induction reduces pulmonary type 2 immune responses, eosinophil activation and alleviates lung lacunarity. In conclusion, we demonstrate that IL-33 signaling is essentially involved in MF-induced ELD development. SUMMARY: Our study demonstrates that repeated sensitization of BALB/c mice with L. sigmodontis MF induces pulmonary eosinophilia in an IL-33-dependent manner. The newly established model recapitulates the characteristic features known to occur during eosinophilic lung diseases (ELD) such as human tropical pulmonary eosinophilia (TPE), which includes the retention of microfilariae in the lung tissue and induction of pulmonary eosinophilia and type 2 immune responses. Our study provides compelling evidence that IL-33 drives eosinophil activation during ELD and that blocking IL-33 signaling using HpARI2 reduces eosinophil activation, eosinophil accumulation in the lung tissue, suppresses type 2 immune responses and mitigates the development of structural damage to the lung. Consequently, IL-33 is a potential therapeutic target to reduce eosinophil-mediated pulmonary pathology.

Differences of in vitro immune responses between patent and pre-patent Litomosoides sigmodontis-infected mice are independent of the filarial antigenic stimulus used.

Lymphatic filariasis and onchocerciasis are neglected tropical diseases and cause significant public health problems in endemic countries, especially in sub-Saharan Africa. Since the human parasites are not viable in immune-competent mice, animal models have been developed, among them Litomosoides sigmodontis which permits a complete life cycle in BALB/c mice, including the development of patent infections with circulating microfilariae (Mf, the worm's offspring). To investigate the immunomodulatory properties of helminths in vitro, antigenic extracts can be prepared from different life cycle stages of the L. sigmodontis model, including adult worms, but the methods to prepare these antigens differ between research groups. This study analyzed whether different centrifugation methods during the preparation of an antigenic extract, the gender of used worms, or the different fractions (soluble or pellet) altered filarial-specific CD4+ T cell responses. These cells were isolated from pre-patent or patent/chronic infected mice, hence those without and those with Mf, respectively. Ex vivo immune responses were compared at these two different time points of the infection as well as the parasitic parameters. Worm burden and cell infiltration were elevated in the thoracic cavity (TC) and draining mediastinal lymph nodes at the pre-patent stage. Within the TC, eosinophils were significantly up-regulated at the earlier time point of infection which was further reflected by the eosinophil-related eotaxin-1 levels. Regarding the production of cytokines by re-stimulated CD4+ T cells in the presence of different antigen preparations, cytokine levels were comparable for all used extracts. Our data show that immune responses differ between pre-patent and patent filarial infection, but not in response to the different antigenic extracts themselves.

[Families with infants and toddlers in the COVID-19 pandemic: parental stress and child development considering social class]. / Familien mit Säuglingen und Kleinkindern in der COVID-19-Pandemie: Belastungserleben und Entwicklungsauffälligkeiten unter Berücksichtigung der sozialen Lage.

INTRODUCTION: It is now well established empirically that families and children who could not attend educational and childcare institutions during the COVID-19 pandemic experienced disadvantages. This is particularly true for families in poverty. However, little is known about the situation of families with young children. The aim of the paper is to investigate (1) to what extent families with infants and toddlers also experienced their situation during the pandemic as stressful, (2) whether there were differences depending on social class, (3) how the pandemic affected the healthy development of infants and toddlers, and (4) to what extent class-related differences can also be identified in this. METHODS: The German National Centre for Early Prevention conducted a nationally representative survey of families with children of age 0 to 3 years in April to December 2022, "Kinder in Deutschland 0­3 2022" (N = 7821). The KiD 0­3 study combines a parent survey on family psychosocial burden and resources with pediatric documentation of child development. RESULTS: Parents with very young children experienced their situation in the COVID-19 pandemic as stressful. A clear difference depending on social class emerged. Both from the parents' perspective and in pediatric judgment, the pandemic had a negative impact on social and affective development even in young children. These effects were more pronounced in children from families experiencing poverty than in children from families not receiving basic government benefits. DISCUSSION: In order to mitigate the psychosocial consequences of the COVID-19 pandemic for families and to improve children's chances of growing up in a healthy way that promotes their development, it is necessary to support families in need without stigmatization.

[An urban-rural comparison of family stress and early childhood intervention: Data linkage between two cross-sectional studies]. / Ein Stadt-Land-Vergleich familialer Belastungen und Früher Hilfen.

An urban-rural comparison of family stress and early childhood intervention: Data linkage between two cross-sectional studies Abstract. Background: So far, there is insufficient information about where in Germany which families are reached by home-visiting programs and how well. One example is the provision of home-visiting program by health service professionals (LaB) in early childhood intervention. Research questions: Are there urban-rural differences in psychosocial stress among families? What differences exist in terms of expanding support services, receiving a service, and using the offered LaB? Methods: Data from a representative family survey on psychosocial stress characteristics and the use of services, and a nationwide survey of professionals on the development and expansion of early childhood intervention at the community level were linked. Analyses included descriptive statistics and regression models. Results: Most psychosocial stress characteristics were reported with similar rates in rural and urban areas. Although the LaB program is more widespread in urban regions, it was more likely to be used in rural regions when offered. LaB was more likely to be used by families with migration background, with signs of depression or anxiety of parents, and with a child with perinatal adversities, and less likely when there was a need for expansion of this service. Conclusions: The increased use in rural areas could be due to the fact that there are long distances to few centralized services. Therefore, the outreach character of the LaB program is appreciated, especially among less mobile families.

[Early childhood intervention from a distance-opportunities and challenges in supporting psychosocially stressed families in the COVID-19 pandemic]. / Frühe Hilfen aus der Distanz ­ Chancen und Herausforderungen bei der Unterstützung psychosozial belasteter Familien in der COVID-19-Pandemie.

As of spring 2020, contact-restricting measures were necessary to contain the incidence of infection with the SARS-CoV­2 virus in Germany. However, these restrictions posed particular challenges to families, especially families in high-stress situations. This paper examines how the COVID-19 pandemic affected the life of families and if long-term support in the context of early childhood intervention could be continued. The following analyses are based on qualitative research with psychosocially stressed mothers of young children, two surveys of health professionals who provide long-term support to families in early childhood intervention, and a survey of municipal actors who are responsible for the management and implementation of early childhood intervention in German communities.During the pandemic, families in high-stress conditions report existential fears and general overstrain. Intrafamilial conflicts seem to increase. Home visits in the context of long-term psychosocial support were initially primarily replaced by phone calls, video calls or other forms of "support from a distance." During the course of the pandemic, many professionals have begun conducting home visiting again, but several elements of support from a distance continue to be used as a supplement.Due to the pandemic-related additional burdens on the families who were accompanied by early childhood intervention, the continuation of support was even more important than it already was. Having been tested by necessity during the pandemic, several formats of support from a distance may have the potential to complement the usual assistance provided by health professionals in early childhood intervention and thus contribute to its quality development push.

Filarial extract of Litomosoides sigmodontis induces a type 2 immune response and attenuates plaque development in hyperlipidemic ApoE-knockout mice.

A type 1 immune response is involved in atherosclerosis progression, whereas the role of a type 2 polarization, especially with regard to an enhanced T helper (Th)2 cell differentiation, is still unclear. Helminths trigger type 2 immune responses, protecting the host from inflammatory disorders. We investigated whether an increased type 2 polarization by administration of Litomosoides sigmodontis adult worm extract (LsAg) affects atherosclerosis in apolipoprotein E-deficient (ApoE-/-) mice. Injections of 50 µg LsAg, i.p. into ApoE-/- mice induced a type 2 immune response shown by increased frequencies of peritoneal eosinophils and alternatively activated macrophages. To analyze the effect of LsAg on atherosclerosis initiation, ApoE-/- mice received a high-fat diet for 12 wk and weekly injections of 50 µg LsAg from wk 5 to 12. Therapeutic effects on advanced atherosclerosis were analyzed in mice that were fed a high-fat diet for 12 wk followed by 12 wk of normal chow and weekly LsAg injections. Both preventive and therapeutic LsAg application significantly decreased plaque size. Therapeutic treatment even caused regression of plaque size and macrophage density in the aortic root and reduced Th1-specific gene expression and intraplaque inflammation. In addition, plaque size after therapeutic treatment was inversely correlated with plaque-infiltrated alternatively activated macrophages. In vitro, LsAg treatment of HUVECs reduced intracellular levels of phosphorylated NF-κB-p65, IκB-α, and JNK1/2. In bifurcation flow-through slides, THP-1 cell adhesion to a HUVEC monolayer was decreased by LsAg in regions of nonuniform shear stress. Applying inhibitors of the respective kinases suggests JNK1/2 inhibition is involved in the suppressed cell adhesion. A switch to an enhanced type 2 immune response by LsAg exerts antiatherogenic effects on murine plaque development, indicating a protective role of a hampered type 1 polarization. In vitro, LsAg affects endothelial signaling pathways, among which JNK1/2 inhibition seems to be involved in the suppression of monocytic cell adhesion under proatherogenic shear stress.-Constanze, K., Tauchi, M., Furtmair, R., Urschel, K., Raaz-Schrauder, D., Neumann, A.-L., Frohberger, S. J., Hoerauf, A., Regus, S., Lang, W., Sagban, T. A., Stumpfe, F. M., Achenbach, S., Hübner, M. P., Dietel, B. Filarial extract of Litomosoides sigmodontis induces a type 2 immune response and attenuates plaque development in hyperlipidemic ApoE-knockout mice.

Cloning and plant-based production of antibody MC10E7 for a lateral flow immunoassay to detect [4-arginine]microcystin in freshwater.

Antibody MC10E7 is one of a small number of monoclonal antibodies that bind specifically to [Arg4]-microcystins, and it can be used to survey natural water sources and food samples for algal toxin contamination. However, the development of sensitive immunoassays in different test formats, particularly user-friendly tests for on-site analysis, requires a sensitive but also cost-effective antibody. The original version of MC10E7 was derived from a murine hybridoma, but we determined the sequence of the variable regions using the peptide mass-assisted cloning strategy and expressed a scFv (single-chain variable fragment) format of this antibody in yeast and a chimeric full-size version in leaves of Nicotiana tabacum and Nicotiana benthamiana to facilitate inexpensive and scalable production. The specific antigen-binding activity of the purified antibody was verified by surface plasmon resonance spectroscopy and ELISA, confirming the same binding specificity as its hybridoma-derived counterpart. The plant-derived antibody was used to design a lateral flow immunoassay (dipstick) for the sensitive detection of [Arg4]-microcystins at concentrations of 100-300 ng/L in freshwater samples collected at different sites. Plant-based production will likely reduce the cost of the antibody, currently the most expensive component of the dipstick immunoassay, and will allow the development of further antibody-based analytical devices and water purification adsorbents for the efficient removal of toxic contaminants.

[The health sector and early childhood intervention: intersectoral collaboration in research]. / Gesundheit und Frühe Hilfen: Die intersektorale Kooperation im Blick der Forschung.

BACKGROUND: Early childhood interventions are locally and regionally organized support services for families from pregnancy until the end of the third year of life. The interventions promote diverse measures to enhance parental skills in order to improve developmental and living circumstances. One crucial element of early childhood intervention in Germany are prevention networks at municipal level. The collaboration of healthcare professionals and child and youth welfare professionals in these networks aims to provide nonstigmatizing access to early childhood intervention for families with psychosocial burdens. From the point of view of the healthcare sector, the research program Together for Families (ZuFa Monitoring) of the National Centre on Early Prevention (NZFH) at the Federal Centre for Health Education (BZgA) has collected representative data at the interfaces of gynecology, obstetrics, pediatrics, and early childhood intervention since 2017. GOAL: The background and goals, as well as design and methods of the ZuFa Monitoring studies are described. For obstetrics clinics and resident pediatricians, sample descriptions, including data on representativeness and early data regarding collaboration quality, are given. EARLY RESULTS: The samples are representative for the population of obstetric clinics and resident pediatricians in Germany. At least two-thirds of the respondents indicate that the proportion of families with psychosocial burdens has increased. Care for psychosocially burdened families is regarded as challenging due to a lack of time, limited financial compensation, and aggravating conditions, such as language barriers. Respondents expect early childhood intervention to alleviate their daily work. DISCUSSION: ZuFa Monitoring collects data regarding the care for families with psychosocial burdens at the interface of the health and the child and youth welfare sector. The research program generates information on inhibitory as well as promoting factors, thereby informing the further development and expansion of prevention networks at the municipal level, and heightening the quality of care for families in the health sector.

Global Genomic Diversity of Human Papillomavirus 11 Based on 433 Isolates and 78 Complete Genome Sequences.

UNLABELLED: Human papillomavirus 11 (HPV11) is an etiological agent of anogenital warts and laryngeal papillomas and is included in the 4-valent and 9-valent prophylactic HPV vaccines. We established the largest collection of globally circulating HPV11 isolates to date and examined the genomic diversity of 433 isolates and 78 complete genomes (CGs) from six continents. The genomic variation within the 2,800-bp E5a-E5b-L1-upstream regulatory region was initially studied in 181/207 (87.4%) HPV11 isolates collected for this study. Of these, the CGs of 30 HPV11 variants containing unique single nucleotide polymorphisms (SNPs), indels (insertions or deletions), or amino acid changes were fully sequenced. A maximum likelihood tree based on the global alignment of 78 HPV11 CGs (30 CGs from our study and 48 CGs from GenBank) revealed two HPV11 lineages (lineages A and B) and four sublineages (sublineages A1, A2, A3, and A4). HPV11 (sub)lineage-specific SNPs within the CG were identified, as well as the 208-bp representative region for CG-based phylogenetic clustering within the partial E2 open reading frame and noncoding region 2. Globally, sublineage A2 was the most prevalent, followed by sublineages A1, A3, and A4 and lineage B. IMPORTANCE: This collaborative international study defined the global heterogeneity of HPV11 and established the largest collection of globally circulating HPV11 genomic variants to date. Thirty novel complete HPV11 genomes were determined and submitted to the available sequence repositories. Global phylogenetic analysis revealed two HPV11 variant lineages and four sublineages. The HPV11 (sub)lineage-specific SNPs and the representative region identified within the partial genomic region E2/noncoding region 2 (NCR2) will enable the simpler identification and comparison of HPV11 variants worldwide. This study provides an important knowledge base for HPV11 for future studies in HPV epidemiology, evolution, pathogenicity, prevention, and molecular assay development.

Chronic filarial infection provides protection against bacterial sepsis by functionally reprogramming macrophages.

Helminths immunomodulate their hosts and induce a regulatory, anti-inflammatory milieu that prevents allergies and autoimmune diseases. Helminth immunomodulation may benefit sepsis outcome by preventing exacerbated inflammation and severe pathology, but the influence on bacterial clearance remains unclear. To address this, mice were chronically infected with the filarial nematode Litomosoides sigmodontis (L.s.) and the outcome of acute systemic inflammation caused by i.p. Escherichia coli injection was determined. L.s. infection significantly improved E. coli-induced hypothermia, bacterial clearance and sepsis survival and correlated with reduced concentrations of associated pro-inflammatory cytokines/chemokines and a less pronounced pro-inflammatory macrophage gene expression profile. Improved sepsis outcome in L.s.-infected animals was mediated by macrophages, but independent of the alternatively activated macrophage subset. Endosymbiotic Wolbachia bacteria that are present in most human pathogenic filariae, as well as L.s., signal via TLR2 and modulate macrophage function. Here, gene expression profiles of peritoneal macrophages from L.s.-infected mice revealed a downregulation of genes involved in TLR signaling, and pulsing of macrophages in vitro with L.s. extract reduced LPS-triggered activation. Subsequent transfer improved sepsis outcome in naïve mice in a Wolbachia- and TLR2-dependent manner. In vivo, phagocytosis was increased in macrophages from L.s.-infected wild type, but not TLR2-deficient animals. In association, L.s. infection neither improved bacterial clearance in TLR2-deficient animals nor ameliorated E. coli-induced hypothermia and sepsis survival. These results indicate that chronic L.s. infection has a dual beneficial effect on bacterial sepsis, reducing pro-inflammatory immune responses and improving bacterial control. Thus, helminths and their antigens may not only improve the outcome of autoimmune and allergic diseases, but may also present new therapeutic approaches for acute inflammatory diseases that do not impair bacterial control.

Membrane Sterols Modulate the Binding Mode of Amphotericin B without Affecting Its Affinity for a Lipid Bilayer.

Membrane-active antibiotics are known to selectively target certain pathogens based on cell membrane properties, such as fluidity, lipid ordering, and phase behavior. These are in turn modulated by the composition of a lipid bilayer and in particular by the presence and type of membrane sterols. Amphotericin B (AmB), the golden standard of antifungal treatment, exhibits higher activity toward ergosterol-rich fungal membranes, which permits its use against systemic mycoses; however, the selectivity for fungal membranes is far from satisfactory leading to severe side effects. Despite decades of research, no consensus has emerged on the origin of AmB specificity for fungal cells and its actual mode of action at the molecular level. Previously, it has been proposed that the specific action of AmB is related to differences in its affinity for membranes of different composition. In this work, we investigate this relationship by employing molecular dynamics simulations to compare the free energy of insertion of AmB into three types of membranes: a pure DMPC bilayer and DMPC bilayers containing 30% of cholesterol or ergosterol. We analyze the orientation of AmB molecules within the bilayer in order to unambiguously establish their membrane binding mode and relate the orientational freedom to the sterol-dependent tightness of lipid packing. Our results strongly indicate that the membrane insertion of AmB proceeds virtually to completion independent of membrane type, and hence the higher toxicity against fungal membranes may rather result from differences in subsequent oligomerization in the membrane and assembly of monomers into functional transmembrane pores. In particular, the latter could be facilitated by sterol-induced ordering of AmB molecules along the membrane normal, revealed by our free energy profiles. Moreover--in contrast to certain claims--we find no stable binding mode corresponding to the horizontal adsorption of AmB on the membrane surface.

[Obstacles for the use of early childhood prevention services : The role of expected ability to control sources]. / Barrieren für die Inanspruchnahme Früher Hilfen : Die Rolle der elterlichen Steuerungskompetenz.

BACKGROUND: It is well established that preventive measures for pregnant women and families with small children do not reach all families alike. Often enough, it is those families, who due to heightened stresses and strains might gain the most from these measures, who cannot be reached (dilemma of prevention). GOAL: This investigation explores whether parents can be grouped according to their views on health, prevention and measures available to young families, and whether belonging to one of the identified groups explains differences in the use of (primary and secondary) preventive measures for pregnant women and young families. METHOD: In the context of home visits, parents of children aged two to four years (N = 273) completed questionnaires. Additionally, 203 parents took part in qualitative interviews. RESULTS AND DISCUSSION: Based on interview data, parents from different sociocultural backgrounds could be grouped along their expected ability to control sources for parental support (Steuerungskompetenz). Parents high and low in Steuerungskompetenz differ regarding their knowledge of, use of, and satisfaction with (primary) preventive measures. Steuerungskompetenz explains the use of primary preventive measures above and beyond socioeconomic control variables as well as family stresses and strains. The use of secondary preventive measures is better explained by family stresses and strains. Results are discussed in terms of untapped services, needs and possibilities of target group-oriented approach.

[Knowledge and use of different support programs in the context of early prevention in relation to family-related psychosocial burden]. / Inanspruchnahme von Angeboten der Frühen Hilfen und darüber hinaus durch psychosozial belastete Eltern.

BACKGROUND: The Federal Initiative for Early Prevention (funded by German ministry BMFSFJ), through the development of specific assistance programmes, supports families that suffer from psychosocial burden. As nationally representative data are missing, the National Centre for Early Prevention carried out a national survey on the psychosocial burden experienced by families with children aged 0-3 years. AIMS: Ascertainment of the connections between family-related psychosocial burden and knowledge and use of different assistance programmes. DATA AND METHODS: Via paediatricians, 8063 parents were recruited to complete a questionnaire on objective burden, subjective experience of burden as well as knowledge and use of assistance programmes. Differences in knowledge and use between educational groups were tested by means of chi-squared tests. Very good knowledge of available assistance programmes and the offer and acceptance of aid by family midwives were subjected to regression analyses. RESULTS: Clear differences in knowledge and use of individual assistance programmes between educational groups were observed. Many programmes are predominantly used by better educated families, although there are exceptions, for example in the case of family midwives. Despite generally small group differences, less-educated families are the proportionally largest user group of family midwives. Furthermore we present average predicted percentages of knowledge and use for specific groups of psychosocially burdened parents as derived from the regression analyses. DISCUSSION: The results are discussed in the context of barriers to access for individual assistance programmes as well as their match with families' needs in the practice of early prevention.

Different utilization of alginate and other algal polysaccharides by marine Alteromonas macleodii ecotypes.

The marine bacterium Alteromonas macleodii is a copiotrophic r-strategist, but little is known about its potential to degrade polysaccharides. Here, we studied the degradation of alginate and other algal polysaccharides by A. macleodii strain 83-1 in comparison to other A. macleodii strains. Cell densities of strain 83-1 with alginate as sole carbon source were comparable to those with glucose, but the exponential phase was delayed. The genome of 83-1 was found to harbour an alginolytic system comprising five alginate lyases, whose expression was induced by alginate. The alginolytic system contains additional CAZymes, including two TonB-dependent receptors, and is part of a 24 kb genomic island unique to the A. macleodii 'surface clade' ecotype. In contrast, strains of the 'deep clade' ecotype contain only a single alginate lyase in a separate 7 kb island. This difference was reflected in an eightfold greater efficiency of surface clade strains to grow on alginate. Strain 83-1 furthermore hydrolysed laminarin, pullulan and xylan, and corresponding polysaccharide utilization loci were detected in the genome. Alteromonas macleodii alginate lyases were predominantly detected in Atlantic Ocean metagenomes. The demonstrated hydrolytic capacities are likely of ecological relevance and represent another level of adaptation among A. macleodii ecotypes.

Global genomic diversity of human papillomavirus 6 based on 724 isolates and 190 complete genome sequences.

UNLABELLED: Human papillomavirus type 6 (HPV6) is the major etiological agent of anogenital warts and laryngeal papillomas and has been included in both the quadrivalent and nonavalent prophylactic HPV vaccines. This study investigated the global genomic diversity of HPV6, using 724 isolates and 190 complete genomes from six continents, and the association of HPV6 genomic variants with geographical location, anatomical site of infection/disease, and gender. Initially, a 2,800-bp E5a-E5b-L1-LCR fragment was sequenced from 492/530 (92.8%) HPV6-positive samples collected for this study. Among them, 130 exhibited at least one single nucleotide polymorphism (SNP), indel, or amino acid change in the E5a-E5b-L1-LCR fragment and were sequenced in full. A global alignment and maximum likelihood tree of 190 complete HPV6 genomes (130 fully sequenced in this study and 60 obtained from sequence repositories) revealed two variant lineages, A and B, and five B sublineages: B1, B2, B3, B4, and B5. HPV6 (sub)lineage-specific SNPs and a 960-bp representative region for whole-genome-based phylogenetic clustering within the L2 open reading frame were identified. Multivariate logistic regression analysis revealed that lineage B predominated globally. Sublineage B3 was more common in Africa and North and South America, and lineage A was more common in Asia. Sublineages B1 and B3 were associated with anogenital infections, indicating a potential lesion-specific predilection of some HPV6 sublineages. Females had higher odds for infection with sublineage B3 than males. In conclusion, a global HPV6 phylogenetic analysis revealed the existence of two variant lineages and five sublineages, showing some degree of ethnogeographic, gender, and/or disease predilection in their distribution. IMPORTANCE: This study established the largest database of globally circulating HPV6 genomic variants and contributed a total of 130 new, complete HPV6 genome sequences to available sequence repositories. Two HPV6 variant lineages and five sublineages were identified and showed some degree of association with geographical location, anatomical site of infection/disease, and/or gender. We additionally identified several HPV6 lineage- and sublineage-specific SNPs to facilitate the identification of HPV6 variants and determined a representative region within the L2 gene that is suitable for HPV6 whole-genome-based phylogenetic analysis. This study complements and significantly expands the current knowledge of HPV6 genetic diversity and forms a comprehensive basis for future epidemiological, evolutionary, functional, pathogenicity, vaccination, and molecular assay development studies.

Cyclin D1 is a strong prognostic factor for survival in pancreatic cancer: analysis of CD G870A polymorphism, FISH and immunohistochemistry.

BACKGROUND AND OBJECTIVE: Cyclin D1 is an important regulator protein for the G1-S cell cycle phase transition. The aim of this trial was to evaluate the impact of the CCND1 polymorphism G870A and corresponding protein expression and CCND1 amplification on the survival of the patients. METHODS: 425 patients with ductal pancreatic adenocarcinoma who underwent resection were included after histopathological confirmation. DNA was analyzed for Cyclin D1 polymorphisms, immunhistochemical examination and fluorescence in situ hybridization analysis of the tumor were performed. RESULTS: Overall, the mean survival was 22.9 months (20.5-25.3). The survival in patients with Cyclin D1 G870A polymorphism Adenine/Adenine was 15.1 months (95% CI 11.3-18.9), 21.5 months (17.4-25.6) for Adenine/Guanine, and 29.4 months (95% CI 23.8-35.0) for Guanine/Guanine (P = 0.003). A shorter survival was associated with strong/moderate protein expression in immunohistochemistry (IHC) compared to weak/no expression (P = 0.028). Additionally, a significant coherency between unfavourable polymorphism (AA/AG) and increased protein expression was detected (P = 0.005). CONCLUSIONS: A strong impact on survival of Cyclin D1 G870A polymorphism and the detected corresponding protein expression was found. The biological mechanism of CCND1 in carcinogenesis has not been fully examined; but at present Cyclin D1 seems to be an interesting biomarker for the prognosis of ductal adenocarcinoma.

Concerted neuron-astrocyte gene expression declines in aging and schizophrenia.

Human brains vary across people and over time; such variation is not yet understood in cellular terms. Here we describe a striking relationship between people's cortical neurons and cortical astrocytes. We used single-nucleus RNA-seq to analyze the prefrontal cortex of 191 human donors ages 22-97 years, including healthy individuals and persons with schizophrenia. Latent-factor analysis of these data revealed that in persons whose cortical neurons more strongly expressed genes for synaptic components, cortical astrocytes more strongly expressed distinct genes with synaptic functions and genes for synthesizing cholesterol, an astrocyte-supplied component of synaptic membranes. We call this relationship the Synaptic Neuron-and-Astrocyte Program (SNAP). In schizophrenia and aging - two conditions that involve declines in cognitive flexibility and plasticity 1,2 - cells had divested from SNAP: astrocytes, glutamatergic (excitatory) neurons, and GABAergic (inhibitory) neurons all reduced SNAP expression to corresponding degrees. The distinct astrocytic and neuronal components of SNAP both involved genes in which genetic risk factors for schizophrenia were strongly concentrated. SNAP, which varies quantitatively even among healthy persons of similar age, may underlie many aspects of normal human interindividual differences and be an important point of convergence for multiple kinds of pathophysiology.

The effect of sterols on amphotericin B self-aggregation in a lipid bilayer as revealed by free energy simulations.

Amphotericin B (AmB) is an effective but toxic antifungal drug, known to increase the permeability of the cell membrane, presumably by assembling into transmembrane pores in a sterol-dependent manner. The aggregation of AmB molecules in a phospholipid bilayer is, thus, crucial for the drug's activity. To provide an insight into the molecular nature of this process, here, we report an atomistic molecular dynamics simulation study of AmB head-to-head dimerization in a phospholipid bilayer, a possible early stage of aggregation. To compare the effect of sterols on the thermodynamics of aggregation and the architecture of the resulting AmB-AmB complexes, free energy profiles for the dimerization in ergosterol- or cholesterol-containing and sterol-free membranes are derived from the simulations. These profiles demonstrate that although AmB dimers are formed in all the systems studied, they are significantly less favorable in the bilayer with ergosterol than in the cholesterol-containing or sterol-free ones. We investigate the structural and energetic determinants of this difference and discuss its consequences for the AmB mechanism of action.

How are families in Germany doing in 2022? Study protocol of a nationally representative, cross-sectional study of parents with children aged 0-3 years.

BACKGROUND: In order for Early Childhood Intervention (ECI) to be effective, data-based information on families' resources, burden and current use of support services for families with young children, as well as on children's health and development is needed. The study Kinder in Deutschland [Children in Germany]-KiD 0-3 2022 aims at providing these data to help us understand families' situation and needs in Germany now, including families' experience of the COVID-19 pandemic. METHOD: The study will recruit up to 300 pediatricians who will invite parents of children aged up to 48 months to participate in the study during a well-child visit. Parents (goal N = 8,000) will complete an online-questionnaire with their own web-enabled device. Pediatricians will complete a short questionnaire about each participating family. The questionnaires cover family psychosocial burden and resources, child health and development, use of family support services, as well as the families' experiences of the COVID-19 pandemic. Data will be analyzed to assess patterns of families´ psychosocial burdens and resources, use of support services for families with young children, and children´s health and development. Concordance between parent and pediatrician report will be assessed and comparisons with the predecessor study of 2015 will be drawn. DISSEMINATION: Findings will be disseminated through scientific conferences, open access peer-reviewed journals, and dissemination channels of the National Centre for Early Prevention. DISCUSSION: The present study will provide parent and pediatrician reports on how families with young children are doing in Germany. These data will be used to inform Germany's early childhood intervention (ECI) program ("Frühe Hilfen") on current needs of families with young children.