An atlas of combinatorial transcriptional regulation in mouse and man.

Combinatorial interactions among transcription factors are critical to directing tissue-specific gene expression. To build a global atlas of these combinations, we have screened for physical interactions among the majority of human and mouse DNA-binding transcription factors (TFs). The complete networks contain 762 human and 877 mouse interactions. Analysis of the networks reveals that highly connected TFs are broadly expressed across tissues, and that roughly half of the measured interactions are conserved between mouse and human. The data highlight the importance of TF combinations for determining cell fate, and they lead to the identification of a SMAD3/FLI1 complex expressed during development of immunity. The availability of large TF combinatorial networks in both human and mouse will provide many opportunities to study gene regulation, tissue differentiation, and mammalian evolution.

The changing nature of HPV associated with high grade cervical lesions in vaccinated populations, a retrospective study of over 1700 cases in Scotland.

BACKGROUND: Understanding the pattern and dominance of HPV types in high grade cervical disease within increasingly vaccinated populations will help inform the development of appropriate screening and management protocols. METHODS: Over 1700 cases of cervical intraepithelial neoplasia (CIN) diagnosed between 2011 and 2017 in women younger than 25 were genotyped for HPV. Logistic regression was used to assess the association between HPV 16/18 positivity with biopsy-collection year, birth year, deprivation and vaccination status. Regression analysis was repeated for cross-protective types (31, 33 and 45). Type specific detail of non-vaccine types by vaccination status was presented descriptively. RESULTS: Detection of HPV 16/18 or 16/18/31/33 and 45 was lower in CIN2 associated with full vaccination vs no vaccination (OR 0.3; 95% CI 0.2-0.5 & 0.4; 95% CI 0.3-0.6 respectively) Similar observations were made for CIN3. The relative contribution of non-established high-risk types including those considered low risk was greater among vaccinated women with CIN2+ vs unvaccinated women with CIN2+. CONCLUSIONS: The change in HPV distribution in CIN2+ in vaccinated populations is a further marker of vaccine impact. Additionally, the progression rate of CIN2+ in vaccinated populations may be lower given the shift in type distribution. The definition of high grade disease in vaccinated populations may warrant reassessment.

Nutrient sensing modulates malaria parasite virulence.

The lifestyle of intracellular pathogens, such as malaria parasites, is intimately connected to that of their host, primarily for nutrient supply. Nutrients act not only as primary sources of energy but also as regulators of gene expression, metabolism and growth, through various signalling networks that enable cells to sense and adapt to varying environmental conditions. Canonical nutrient-sensing pathways are presumed to be absent from the causative agent of malaria, Plasmodium, thus raising the question of whether these parasites can sense and cope with fluctuations in host nutrient levels. Here we show that Plasmodium blood-stage parasites actively respond to host dietary calorie alterations through rearrangement of their transcriptome accompanied by substantial adjustment of their multiplication rate. A kinome analysis combined with chemical and genetic approaches identified KIN as a critical regulator that mediates sensing of nutrients and controls a transcriptional response to the host nutritional status. KIN shares homology with SNF1/AMPKα, and yeast complementation studies suggest that it is part of a functionally conserved cellular energy-sensing pathway. Overall, these findings reveal a key parasite nutrient-sensing mechanism that is critical for modulating parasite replication and virulence.

Metabolic Potential of the Gut Microbiome Is Significantly Impacted by Conditioning Regimen in Allogeneic Hematopoietic Stem Cell Transplantation Recipients.

Allogeneic hematopoietic stem cell transplantation (aHSCT) is a putative curative treatment for malignant hematologic disorders. During transplantation, the immune system is suppressed/eradicated through a conditioning regimen (non-myeloablative or myeloablative) and replaced with a donor immune system. In our previous study, we showed changes in gut taxonomic profiles and a decrease in bacterial diversity post-transplant. In this study, we expand the cohort with 114 patients and focus on the impact of the conditioning regimens on taxonomic features and the metabolic functions of the gut bacteria. This is, to our knowledge, the first study to examine the metabolic potential of the gut microbiome in this patient group. Adult aHSCT recipients with shotgun sequenced stool samples collected day -30 to +28 relative to aHSCT were included. One sample was selected per patient per period: pre-aHSCT (day -30-0) and post-aHSCT (day 1-28). In total, 254 patients and 365 samples were included. Species richness, alpha diversity, gene richness and metabolic richness were all lower post-aHSCT than pre-aHSCT and the decline was more pronounced for the myeloablative group. The myeloablative group showed a decline in 36 genera and an increase in 15 genera. For the non-myeloablative group, 30 genera decreased and 16 increased with lower fold changes than observed in the myeloablative group. For the myeloablative group, 32 bacterial metabolic functions decreased, and one function increased. For the non-myeloablative group, three functions decreased, and two functions increased. Hence, the changes in taxonomy post-aHSCT caused a profound decline in bacterial metabolic functions especially in the myeloablative group, thus providing new evidence for associations of myeloablative conditioning and gut dysbiosis from a functional perspective.

Clipped histone H3 is integrated into nucleosomes of DNA replication genes in the human malaria parasite Plasmodium falciparum.

Post-translational modifications of histone H3 N-terminal tails are key epigenetic regulators of virulence gene expression and sexual commitment in the human malaria parasite Plasmodium falciparum Here, we identify proteolytic clipping of the N-terminal tail of nucleosome-associated histone H3 at amino acid position 21 as a new chromatin modification. A cathepsin C-like proteolytic clipping activity is observed in nuclear parasite extracts. Notably, an ectopically expressed version of clipped histone H3, PfH3p-HA, is targeted to the nucleus and integrates into mononucleosomes. Furthermore, chromatin immunoprecipitation and next-generation sequencing analysis identified PfH3p-HA as being highly enriched in the upstream region of six genes that play a key role in DNA replication and repair: In these genes, PfH3p-HA demarcates a specific 1.5 kb chromatin island adjacent to the open reading frame. Our results indicate that, in P. falciparum, the process of histone clipping may precede chromatin integration hinting at preferential targeting of pre-assembled PfH3p-containing nucleosomes to specific genomic regions. The discovery of a protease-directed mode of chromatin organization in P. falciparum opens up new avenues to develop new anti-malarials.

"It made me feel brighter in myself"- The health and well-being impacts of a residential front garden horticultural intervention.

Residential gardens make up 30% of urban space in the UK, yet unlike many other green space typologies, their role in the health and well-being agenda has largely been overlooked. A horticultural intervention introduced ornamental plants to 38 previously bare front gardens (≈ 10 m2) within an economically deprived region of North England, UK. Measures of perceived stress and diurnal cortisol profiles (as an indicator of health status) were taken pre- and post-intervention (over 3 months). Residents reported significant decreases in perceived stress post-intervention. This finding was aligned with a higher proportion of 'healthy' diurnal cortisol patterns post-intervention, suggesting better health status in those individuals. All residents derived one or more reported socio-cultural benefits as a result of the front garden plantings, although overall scores for subjective well-being did not increase to a significant level. Further qualitative data suggested that the gardens were valued for enhancing relaxation, increasing positive emotions, motivation, and pride of place. The results indicate that adding even small quantities of ornamental plants to front gardens within deprived urban communities had a positive effect on an individual's stress regulation and some, but not all, aspects of subjective well-being. The research highlights the importance of residential front gardens to human health and well-being, and thus their contribution to the wider debates around city densification, natural capital and urban planning.

Baseline HPV prevalence in rectal swabs from men attending a sexual health clinic in Scotland: assessing the potential impact of a selective HPV vaccination programme for men who have sex with men.

OBJECTIVES: A human papillomavirus (HPV) vaccination programme targeted towards men who have sex with men who are disproportionately affected by HPV anogenital infection and related disease was established in Scotland in July 2017. We aimed to establish a baseline HPV prevalence to assess the potential impact of the programme. METHODS: Residual rectal swabs taken in a sexual health clinic (n=1 248) were tested for the presence of HPV and HPV-type prevalence was collated and stratified by age. Prevalence of HPV types included in the quadrivalent and nonavalent vaccines was specifically assessed. RESULTS: 72.8% (95% CI 70.2% to 75.3%) of swabs were positive for HPV with 59.1% (95% CI 56.3% to 61.9%) of samples positive for at least one high-risk type. A least one of HPV 6, 11, 16 and 18 was detected in approximately half of the swabs. HPV prevalence generally increased with age but did not significantly differ between older age groups. The presence of more than one HPV type increased with age and over half of samples had multiple types present. CONCLUSIONS: While HPV prevalence in this population is high, the potential impact of the vaccination programme is substantial given that 50% are not infected with a vaccine type. Defining a preimmunisation baseline in this group will be important for longitudinal monitoring of impact.

Association Between Single-Nucleotide Polymorphisms in HLA Alleles and Human Immunodeficiency Virus Type 1 Viral Load in Demographically Diverse, Antiretroviral Therapy-Naive Participants From the Strategic Timing of AntiRetroviral Treatment Trial.

The impact of variation in host genetics on replication of human immunodeficiency virus type 1 (HIV-1) in demographically diverse populations remains uncertain. In the current study, we performed a genome-wide screen for associations of single-nucleotide polymorphisms (SNPs) to viral load (VL) in antiretroviral therapy-naive participants (n = 2440) with varying demographics from the Strategic Timing of AntiRetroviral Treatment (START) trial. Associations were assessed using genotypic data generated by a customized SNP array, imputed HLA alleles, and multiple linear regression. Genome-wide significant associations between SNPs and VL were observed in the major histocompatibility complex class I region (MHC I), with effect sizes ranging between 0.14 and 0.39 log10 VL (copies/mL). Supporting the SNP findings, we identified several HLA alleles significantly associated with VL, extending prior observations that the (MHC I) is a major host determinant of HIV-1 control with shared genetic variants across diverse populations and underscoring the limitations of genome-wide association studies as being merely a screening tool.

Identification of the centromeres of Leishmania major: revealing the hidden pieces.

Leishmania affects millions of people worldwide. Its genome undergoes constitutive mosaic aneuploidy, a type of genomic plasticity that may serve as an adaptive strategy to survive distinct host environments. We previously found high rates of asymmetric chromosome allotments during mitosis that lead to the generation of such ploidy. However, the underlying molecular events remain elusive. Centromeres and kinetochores most likely play a key role in this process, yet their identification has failed using classical methods. Our analysis of the unconventional kinetochore complex recently discovered in Trypanosoma brucei (KKTs) leads to the identification of a Leishmania KKT gene candidate (LmKKT1). The GFP-tagged LmKKT1 displays "kinetochore-like" dynamics of intranuclear localization throughout the cell cycle. By ChIP-Seq assay, one major peak per chromosome is revealed, covering a region of 4 ±2 kb. We find two largely conserved motifs mapping to 14 of 36 chromosomes while a higher density of retroposons are observed in 27 of 36 centromeres. The identification of centromeres and of a kinetochore component of Leishmania chromosomes opens avenues to explore their role in mosaic aneuploidy.

Moving epidemic method (MEM) applied to virology data as a novel real time tool to predict peak in seasonal influenza healthcare utilisation. The Scottish experience of the 2017/18 season to date.

Scotland observed an unusual influenza A(H3N2)-dominated 2017/18 influenza season with healthcare services under significant pressure. We report the application of the moving epidemic method (MEM) to virology data as a tool to predict the influenza peak activity period and peak week of swab positivity in the current season. This novel MEM application has been successful locally and is believed to be of potential use to other countries for healthcare planning and building wider community resilience.

Increase in Legionnaires' disease cases associated with travel to Dubai among travellers from the United Kingdom, Sweden and the Netherlands, October 2016 to end August 2017.

Between 1 October 2016 and 31 August 2017, 51 Legionnaires' disease (LD) cases from the United Kingdom, Sweden and the Netherlands were identified with associated travel to Dubai. Cases did not all stay in the same accommodation, indicating that no single accommodation could be the source for all these infections. While local investigations continue into other potential sources, clinicians should remain alert to the possibility of LD among travellers returning from Dubai with respiratory illness.

Human Papillomavirus Prevalence and Herd Immunity after Introduction of Vaccination Program, Scotland, 2009-2013.

In 2008, a national human papillomavirus (HPV) immunization program using a bivalent vaccine against HPV types 16 and 18 was implemented in Scotland along with a national surveillance program designed to determine the longitudinal effects of vaccination on HPV infection at the population level. Each year during 2009-2013, the surveillance program conducted HPV testing on a proportion of liquid-based cytology samples from women undergoing their first cervical screening test for precancerous cervical disease. By linking vaccination, cervical screening, and HPV testing data, over the study period we found a decline in HPV types 16 and 18, significant decreases in HPV types 31, 33, and 45 (suggesting cross-protection), and a nonsignificant increase in HPV 51. In addition, among nonvaccinated women, HPV types 16 and 18 infections were significantly lower in 2013 than in 2009. Our results preliminarily indicate herd immunity and sustained effectiveness of the bivalent vaccine on virologic outcomes at the population level.

First efficient CRISPR-Cas9-mediated genome editing in Leishmania parasites.

Protozoan pathogens that cause leishmaniasis in humans are relatively refractory to genetic manipulation. In this work, we implemented the CRISPR-Cas9 system in Leishmania parasites and demonstrated its efficient use for genome editing. The Cas9 endonuclease was expressed under the control of the Dihydrofolate Reductase-Thymidylate Synthase (DHFR-TS) promoter and the single guide RNA was produced under the control of the U6snRNA promoter and terminator. As a proof of concept, we chose to knockout a tandemly repeated gene family, the paraflagellar rod-2 locus. We were able to obtain null mutants in a single round of transfection. In addition, we confirmed the absence of off-target editions by whole genome sequencing of two independent clones. Our work demonstrates that CRISPR-Cas9-mediated gene knockout represents a major improvement in comparison with existing methods. Beyond gene knockout, this genome editing tool opens avenues for a multitude of functional studies to speed up research on leishmaniasis.

Green infrastructure and ecosystem services - is the devil in the detail?

BACKGROUND: Green infrastructure is a strategic network of green spaces designed to deliver ecosystem services to human communities. Green infrastructure is a convenient concept for urban policy makers, but the term is used too generically and with limited understanding of the relative values or benefits of different types of green space and how these complement one another. At a finer scale/more practical level, little consideration is given to the composition of the plant communities, yet this is what ultimately defines the extent of service provision. This paper calls for greater attention to be paid to urban plantings with respect to ecosystem service delivery and for plant science to engage more fully in identifying those plants that promote various services. SCOPE: Many urban plantings are designed based on aesthetics alone, with limited thought on how plant choice/composition provides other ecosystem services. Research is beginning to demonstrate, however, that landscape plants provide a range of important services, such as helping mitigate floods and alleviating heat islands, but that not all species are equally effective. The paper reviews a number of important services and demonstrates how genotype choice radically affects service delivery. CONCLUSIONS: Although research is in its infancy, data are being generated that relate plant traits to specific services, thereby helping identify genotypes that optimize service delivery. The urban environment, however, will become exceedingly bland if future planting is simply restricted to monocultures of a few 'functional' genotypes. Therefore, further information is required on how to design plant communities where the plants identified (1) provide more than a single benefit (multifunctionality), (B) complement each other in maximizing the range of benefits that can be delivered in one location, and (3) continue to maintain public acceptance through diversity. The identification/development of functional landscape plants is an exciting and potentially high-impact arena for plant science.

Assessing real world vaccine effectiveness: A review of Scotland's approach to monitoring human papillomavirus (HPV) vaccine impact on HPV infection and cervical disease.

High-risk human papillomavirus (HPV) infections can progress to cervical cancer which is the fourth most common cancer in women globally. In Scotland, the incidence of cervical cancer has a strong socioeconomic deprivation gradient disproportionately affecting women from more deprived areas. An HPV vaccination programme was initiated in Scotland in 2008 targeting girls aged 12-13 years with a catch-up campaign running for the first three years for girls aged up to 18 years. The programme has evolved over the last 16 years with changes in the type of vaccine, dosing schedules and the extension of the programme to boys and gay, bisexual and other men who have sex with men. Vaccine uptake in Scotland has historically been high but has gradually decreased over time and disparities exist in women from more deprived areas of Scotland. The ability to link national immunisation and screening databases in Scotland has allowed direct monitoring of the impact of the HPV vaccine on virological and histological outcomes. Analyses of this linked data have demonstrated real-world evidence of high vaccine effectiveness against HPV infection, cervical disease, and cervical cancer with evidence of herd immunity in unvaccinated women. Continued monitoring is crucial to assess the duration of protection, the impact of vaccine and dosing schedules changes and the emergence of potential type replacement. With the World Health Organisation's aim to eliminate cervical cancer as a public health problem by the next century addressing the inequalities in cervical cancer incidence will be crucial. This will require targeted interventions for women most at risk of cervical cancer to ensure elimination is achieved timely for all women in Scotland.

Does increasing biodiversity in an urban woodland setting promote positive emotional responses in humans? A stress recovery experiment using 360-degree videos of an urban woodland.

Green spaces can support human stress reduction and foster positive emotional well-being. Previous research has suggested that biodiversity (i.e. the variety of species of plants and animals in a given location) can enhance recovery from stress even further. However, there is limited experimental evidence testing this hypothesis and results, to date, have been mixed. This study aimed to provide further understanding of the role of biodiversity (actual or perceived) on human well-being by experimentally manipulating species richness and stress. Participants (372 in total) took part in an online experiment, where they received an episode of mild stress before watching a 360-degree video to recover. The video showed the same location, an urban woodland, but at one of four artificially manipulated levels of biodiversity. The participants reported their Positive and Negative Affect before and after the stress induction and after watching the video, providing a measure of their stress and well-being throughout the experiment. Participants also reported their perceptions of biodiversity (i.e. how diverse they thought the location was) and elaborated on their responses with brief comments. Repeated Measure Analysis of Variance revealed that exposure to all levels of biodiversity reduced the participants' Negative Affect, but with no significant difference between the conditions. However, the analysis showed higher Positive Affect in those participants who perceived the environment as more biodiverse. Comments from participants indicated that those who reported noticing flowers and trees in the environment also showed higher Positive Affect. This suggests that perceiving biodiversity promotes more positive emotions, but critically one needs to actually notice (engage with) the components of biodiversity to elicit these extra benefits.

Invasive cervical cancer incidence following bivalent human papillomavirus vaccination: a population-based observational study of age at immunization, dose, and deprivation.

BACKGROUND: High-risk human papillomavirus causes cervical cancer. Vaccines have been developed that significantly reduce the incidence of preinvasive and invasive disease. This population-based observational study used linked screening, immunization, and cancer registry data from Scotland to assess the influence of age, number of doses, and deprivation on the incidence of invasive disease following administration of the bivalent vaccine. METHODS: Data for women born between January 1, 1988, and June 5, 1996, were extracted from the Scottish cervical cancer screening system in July 2020 and linked to cancer registry, immunization, and deprivation data. Incidence of invasive cervical cancer per 100 000 person-years and vaccine effectiveness were correlated with vaccination status, age at vaccination, and deprivation; Kaplan Meier curves were calculated. RESULTS: No cases of invasive cancer were recorded in women immunized at 12 or 13 years of age irrespective of the number of doses. Women vaccinated at 14 to 22 years of age and given 3 doses of the bivalent vaccine showed a significant reduction in incidence compared with all unvaccinated women (3.2/100 000 [95% confidence interval (CI) = 2.1 to 4.6] vs 8.4 [95% CI = 7.2 to 9.6]). Unadjusted incidence was significantly higher in women from most deprived (Scottish Index of Multiple Deprivation 1) than least deprived (Scottish Index of Multiple Deprivation 5) areas (10.1/100 000 [95% CI = 7.8 to 12.8] vs 3.9 [95% CI = 2.6 to 5.7]). Women from the most deprived areas showed a significant reduction in incidence following 3 doses of vaccine (13.1/100 000 [95% CI = 9.95 to 16.9] vs 2.29 [95% CI = 0.62 to 5.86]). CONCLUSION: Our findings confirm that the bivalent vaccine prevents the development of invasive cervical cancer and that even 1 or 2 doses 1 month apart confer benefit if given at 12-13 years of age. At older ages, 3 doses are required for statistically significant vaccine effectiveness. Women from more deprived areas benefit more from vaccination than those from less deprived areas.

Applying the concept of niche breadth to understand urban tree mortality in the UK.

Accelerated climate change has raised concerns about heightened vulnerability of urban trees, spurring the need to reevaluate their suitability. The urgency has also driven the widespread application of climatic niche-based models. In particular, the concept of niche breadth (NB), the range of environmental conditions that species can tolerate, is commonly estimated based on species occurrence data over the selected geographic range to predict species response to changing conditions. However, in urban environments where many species are cultivated out of the NB of their natural distributions, additional empirical evidence beyond presence and absence is needed not only to test the true tolerance limits but also to evaluate species' adaptive capacity to future climate. In this research, mortality trends of Acer and Quercus species spanning a 21-year period (2000-2021) from tree inventories of three major UK botanic gardens - the Royal Botanic Gardens, Kew (KEW), Westonbirt, the National Arboretum (WESB), and the Royal Botanic Garden Edinburgh (RBGE) - were analyzed in relation to their NB under long-term drought stress. As a result, Acer species were more responsive to drought and heat stress. For Acer, positioning below the lower limits of the precipitation of warmest quarter led to an increase in the probability of annual mortality by 1.2 and 1.3 % at KEW and RBGE respectively. In addition, the mean cumulative mortality rate increased corresponding to an increase in the number of niche positions below the lower limits of the selected bioclimatic variables. On the other hand, Quercus species in general exhibited comparable resilience regardless of their niche positions. Moreover, Mediterranean oaks were most tolerant, with cumulative mortality rates that were lower than those of native oaks in the UK. These findings further highlight the importance of incorporating ecological performance and recognizing species-specific adaptive strategies in climatic niche modeling.

Development of a machine learning model for early prediction of plasma leakage in suspected dengue patients.

BACKGROUND: At least a third of dengue patients develop plasma leakage with increased risk of life-threatening complications. Predicting plasma leakage using laboratory parameters obtained in early infection as means of triaging patients for hospital admission is important for resource-limited settings. METHODS: A Sri Lankan cohort including 4,768 instances of clinical data from N = 877 patients (60.3% patients with confirmed dengue infection) recorded in the first 96 hours of fever was considered. After excluding incomplete instances, the dataset was randomly split into a development and a test set with 374 (70%) and 172 (30%) patients, respectively. From the development set, five most informative features were selected using the minimum description length (MDL) algorithm. Random forest and light gradient boosting machine (LightGBM) were used to develop a classification model using the development set based on nested cross validation. An ensemble of the learners via average stacking was used as the final model to predict plasma leakage. RESULTS: Lymphocyte count, haemoglobin, haematocrit, age, and aspartate aminotransferase were the most informative features to predict plasma leakage. The final model achieved the area under the receiver operating characteristics curve, AUC = 0.80 with positive predictive value, PPV = 76.9%, negative predictive value, NPV = 72.5%, specificity = 87.9%, and sensitivity = 54.8% on the test set. CONCLUSION: The early predictors of plasma leakage identified in this study are similar to those identified in several prior studies that used non-machine learning based methods. However, our observations strengthen the evidence base for these predictors by showing their relevance even when individual data points, missing data and non-linear associations were considered. Testing the model on different populations using these low-cost observations would identify further strengths and limitations of the presented model.

Metabolic Profiling Early Post-Allogeneic Haematopoietic Cell Transplantation in the Context of CMV Infection.

Immune dysfunction resulting from allogeneic haematopoietic stem cell transplantation (aHSCT) predisposes one to an elevated risk of cytomegalovirus (CMV) infection. Changes in metabolism have been associated with adverse outcomes, and in this study, we explored the associations between metabolic profiles and post-transplantation CMV infection using plasma samples collected 7-33 days after aHSCT. We included 68 aHSCT recipients from Rigshospitalet, Denmark, 50% of whom experienced CMV infection between days 34-100 post-transplantation. First, we investigated whether 12 metabolites selected based on the literature were associated with an increased risk of post-transplantation CMV infection. Second, we conducted an exploratory network-based analysis of the complete metabolic and lipidomic profiles in relation to clinical phenotypes and biological pathways. Lower levels of trimethylamine N-oxide were associated with subsequent CMV infection (multivariable logistic regression: OR = 0.63; 95% CI = [0.41; 0.87]; p = 0.01). Explorative analysis revealed 12 clusters of metabolites or lipids, among which one was predictive of CMV infection, and the others were associated with conditioning regimens, age upon aHSCT, CMV serostatus, and/or sex. Our results provide evidence for an association between the metabolome and CMV infection post-aHSCT that is independent of known risk factors.