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SummaryNumerous studies have identified traits associated with pollen mimicry, however, the processes underlying floral deception remains poorly documented for these structures. We studied the importance of attraction and mechanical fit of anther mimics in Tritonia laxifolia (Iridaceae) and their relative contributions to reproductive success.To determine anther mimics role in pollinator attraction, we offered bees’ binary preferences to flowers painted with UV absorbent and reflecting paint. We also conducted preference experiments between flowers with excised anther mimics and unmanipulated controls, from which mechanical fit was assessed using single visits. Anther mimics effects on female reproductive success was determined using similar treatments, but on rooted plants.Bees preferred UV absorbent over UV reflecting anther mimics. Preference for flowers with and without the three-dimensional structures was equal. Single visits resulted in more pollen deposition on unmanipulated controls over flowers with their anther mimics excised, which was directly linked to pollen-collecting behaviour. Controls with unmanipulated anther mimics experienced more seed set than those with their anther mimics excised.This study provides insights into pollinator-mediated selection on deceptive floral signals and shows that three-dimensional anther mimics increases reproductive success through both attraction and pollen collecting behaviours that improves the fit between flowers and pollinators.

Quantitative CD8+ Counts in patients with Liver and skin metastases.

Univariate Analysis of Pretreatment prognostic factors in the Discovery cohort.

Supplementary Figure 1. Exome sequencing and copy number changes in M431 and corresponding whole-tumor biopsy. Supplementary Figure 2. Mutations in antigen presentation machinery from anti-PD1 treated melanoma cohort. Supplementary Figure 3. Immunohistochemistry (IHC) analysis of biopsies from melanoma metastases with a JAK1 loss of function mutation. Supplementary Figure 4. Selection of PD-L1 flow cytometry antibody and impact of phosphatase inhibitor as well as temperature on measuring surface PD-L1 expression and pAKT. Supplementary Figure 5. Flow cytometry gating strategy and dose response curve of interferon alpha, beta and gamma to determine the optimal concentrations. Supplementary Figure 6. Time course of PD-L1 surface expression upon interferon alpha, beta and gamma treatment for selected cell lines to determine the optimal time point for the screening. Supplementary Figure 7. Time course of PD-L1 expression upon interferon alpha, beta or gamma treatment for the cell lines with poor or no up-regulation upon 18 hours exposure. Supplementary Figure 8. Interferon signaling pathway in good and poorly responding cell lines. Supplementary Figure 9. PD-L1 expression upon interferon alpha and beta exposure. Supplementary Figure 10. Predicted functional consequences of M368 JAK2 D313 splice site mutation. Supplementary Figure 11. Immunohistochemistry (IHC) analysis of biopsies from melanoma metastases with a JAK1 loss of function mutation. Supplementary Figure 12. JAK1 wild-type lentiviral vector transduction. Supplementary Figure 13. Mutations in antigen presentation machinery from anti-PD1 treated colorectal cohort. Supplementary Figure 14. DNA damage repair gene mutations in endometrial cancer cell lines with JAK1/2 mutations. Supplementary Figure 15. Frequency of JAK1 and JAK2 alterations and their association with overall survival in additional TCGA datasets. Supplementary Figure 16. Functional effect of genetic loss of reactive PD-L1 on responses to PD-1 blockade. Supplementary Database 1. Whole exome sequencing of 23 baseline biopsies from patients analyzed in Fig. 1, and whole exome sequencing of the M431 cell line (Accession code: SRP067938).

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Supplemental Figure 1. Schematic presentation of the analyzed parameters in the correlative cohort per progression survival (PFS). TMB: total mutational burden. PR: partial response. SD: stable disease. PD: progressive disease. â-³: mutation. Supplemental Figure 2. Progression Free Survival (PFS) and subgroup survival analysis of the correlative cohort (N=38) by Log-rank (Mantel-Cox) test. PFS By PD-L1 categories ( 0-49% n=23 vs 50-100% n=9, p75th percentile n=6, p=0.142) PFS By CD8 categories (0-5% n=18 vs 6-25% n=14, p=0.037) PFS By CD4 categories (0-5% n=21 vs 6-25% n=9 vs >25% n=2, p=0.123) Supplemental Figure 3. Association among the four correlative parameters. Supplemental Table 1. Association of the four correlative parameters and with other clinical characteristics. Supplemental Table 2. Correlation of patient characteristics and ORR per irRC (PR vs SD/PD). Supplemental Table 3. Patient characteristics (as categorical variables) in the correlative cohort per prior lines of therapy. Supplemental Table 4. PFS and OS of correlative characteristics (as categorical variables) by year. Supplemental Table 5. Characteristics as continuous variables of long term benefiters (Overall survival > 3 years with no intervening therapies) vs all other patients in the correlative cohort.

Purpose:Several biomarkers have been individually associated with response to PD-1 blockade, including PD-L1 and tumor mutational burden (TMB) in non–small cell lung cancer (NSCLC), and CD8 cells in melanoma. We sought to examine the relationship between these distinct variables with response to PD-1 blockade and long-term benefit.Experimental Design:We assessed the association between baseline tumor characteristics (TMB, PD-L1, CD4, and CD8) and clinical features and outcome in 38 patients with advanced NSCLC treated with pembrolizumab (median follow-up of 4.5 years, range 3.8–5.5 years).Results:PD-L1 expression and CD8 infiltration correlated with each other and each significantly associated with objective response rate (ORR) and progression-free survival (PFS). TMB was independent of PD-L1 and CD8 expression, and trended towards association with ORR and PFS. There was no association between CD4 infiltration and outcomes. Only PD-L1 expression was correlated with overall survival (OS). Among 5 patients with long-term survival >3 years with no additional systemic therapy, PD-L1 expression was the only discriminating feature. The increased predictive value for PFS and OS of composite biomarker inclusive of PD-L1, CD8, CD4, and TMB was limited.Conclusions:In patients with NSCLC treated with PD-1 blockade with long-term follow up, TMB, PD-L1, and CD8 were each associated with benefit from PD-1 blockade. Pretreatment PD-L1 expression was correlated with T lymphocyte infiltration and OS, whereas models incorporating TMB and infiltrating CD4 and CD8 lymphocytes did not substantially add to the predictive value of PD-L1 alone for OS.

Orchidaceae show remarkable diversity in pollination strategies, but how these strategies vary globally is not entirely clear. To identify regions and taxa that are data-rich and lend themselves to rigorous analyses or are data-poor and need attention, we introduce a global database of orchid reproductive biology. Our database contains > 2900 species representing all orchid subfamilies and 23 of 24 tribes. We tabulated information on habit, breeding systems, means of pollinator attraction and the identity of pollinators. Patterns of reproductive biology by habit, geography and taxonomy are presented graphically and analysed statistically. On the basis of our database, most orchid species sampled are pollinator dependent (76%) and self-compatible (88%). Pollinator attraction based on rewards occurs in 54% of the species, whereas 46% use some means of deceit. Orchids generally have highly specific pollinator interactions (median number of pollinator species = 1). Nonetheless, on average, specificity is lower for species offering rewards, occurring in multiple continental regions or Northern America (as defined by the Taxonomic Database Working Group Level 1 regions). Although our database reveals impressive knowledge gains, extensive gaps in basic observations of orchid reproductive biology exist, particularly in tropical regions and diverse lineages of fly-pollinated species. The database is expected to facilitate targeted studies, further elucidating the ecological and evolutionary drivers of orchid diversity.

Long-proboscid fly (LPF) pollination has been recorded in multiple plant lineages in South Africa. However in the large and morphologically diverse genus Erica, evidence for LPF pollination is limited to predictions based upon floral syndromes as well as anecdotal observation and limited citizen science records. Here we use breeding system experiments, pollinator exclusion, remote camera traps, spectral analysis and field observation in the Klein River Mountains, Western Cape Province, South Africa, to demonstrate this mode of pollination in the range-restricted taxon Erica aristata. This species has long narrow corolla tubes with narrow opening and contrasting white and crimson pink petals. Flowers were visited and pollinated exclusively by one long-proboscid fly species, Prosoeca rubicunda (Nemestrinidae) over four years at this study site. Breeding experiments showed that E. aristata is not self-compatible and pollinator exclusion experiments indicate that pollinator visits were required for seed-set. Low nectar volume and moderately high nectar sugar concentration were consistent with other long-proboscid fly-pollinated plant species in the Cape. This study represents the first confirmed instance of long-proboscid fly-pollination in the genus Erica.

Cancer cell plasticity due to the dynamic architecture of interactome networks provides a vexing outlet for therapy evasion. Here, through chemical biology approaches for systems level exploration of protein connectivity changes applied to pancreatic cancer cell lines, patient biospecimens, and cell- and patient-derived xenografts in mice, we demonstrate interactomes can be re-engineered for vulnerability. By manipulating epichaperomes pharmacologically, we control and anticipate how thousands of proteins interact in real-time within tumours. Further, we can essentially force tumours into interactome hyperconnectivity and maximal protein-protein interaction capacity, a state whereby no rebound pathways can be deployed and where alternative signalling is supressed. This approach therefore primes interactomes to enhance vulnerability and improve treatment efficacy, enabling therapeutics with traditionally poor performance to become highly efficacious. These findings provide proof-of-principle for a paradigm to overcome drug resistance through pharmacologic manipulation of proteome-wide protein-protein interaction networks., Joshi, Gomes et al. employ a chemical modulation approach of the cellular interactome to a hyperconnectivity state and show association with the increased response of pancreatic cancer cell lines to specific drugs, including those that target the MAPK-pathways and PI3K-mTOR pathway. To achieve this, the authors employ chemical modulation of the interactome via epichaperome inhibition with the small molecule PU-H71.

Background Breastmilk (BM) is a complex fluid that contributes to shaping the immune system of the offspring. BM composition depends on stage of lactation, maternal health status and diet, environment, and genetics. Limited data exists on the composition of the BM from women with IBD and its potential impact on the newborn’s microbiome composition. Methods The MECONIUM (Exploring MEChanisms Of disease traNsmission In Utero through the Microbiome) study is a prospective cohort study including pregnant women with IBD, pregnant healthy control (HC), and their offspring. BM samples were collected 2 weeks post-delivery. Stool samples from the offspring were collected throughout the first 3 years of life and used to assess faecal calprotectin (fCal) and gut microbiota composition (16S). Targeted proteomics of the BM samples was performed with the Olink inflammation panel (92 protein biomarkers). Correlations between specific proteins in the BM, fCal and 16S were assessed using non-parametric tests. Multiple testing correction was performed with false discovery rate (FDR). MaAsLin2 R package was used for multivariate testing. Results 236 BM samples were analysed: 174 from HC, 37 Crohn’s disease (CD), 25 ulcerative colitis (UC). Thymic stromal lymphopoietin (TSLP), a cytokine with an important role in the maturation of T cells, was significantly lower in BM of women with IBD vs HC (FDR p=0.0017). The levels of TSLP in the BM of the mothers correlated negatively with infant fCal at year1 (rho=-0.20, p=0.01), and with the relative abundance of Cronobacter (MaAsLin2 FDR 0.1) of the offspring at month 1. Chemokine (C-C motif) ligand 20 (CCL20), which acts in chemotaxis of dendritic cells and T-cells and B-cells, was also significantly lower in women with CD vs HC (FDR 0.013) and in women with CD vs UC (p=0.014). Matrix metalloproteinase-1 (MMP-1), a collagenase involved in the breakdown of extracellular matrix, was also lower in BM of women with CD (p=0.009) and a negative correlation was observed between the levels of MMP1 and fCal at 3 months and 1 year (rho=-0.20 and -0.18, p=0.01 and 0.02, respectively). Osteoprotegerin (OPG), higher in BM of women with UC (p=0.018), was positively correlated with Streptococcus (MaAsLin2 FDR p=0.2) and negatively correlated with Bacteroides and Parabacteroides (MaAsLin2 FDR p=0.03 and 0.1) in the offspring at month 1. Conclusion The proteomic profile of BM of women with IBD is distinct from that of women without IBD. BM composition may influence offspring’s’ gut microbiome signatures and fCal level at different timepoints. These findings suggest that BM composition may impact the offspring’s intestinal immune system maturation and microbiome development, and warrant further research.

The study area of Cala Gonone in NE Sardinia (Italy) consists of a wide terraced re-entrance/valley crowned inland by carbonate hills and, near the coast bounded laterally and partly floored by thin basaltic lava lying over carbonate bedrock. In this re-entrance, several inland alluvial fans have developed, and a local ~ 30 m high, about 10 m wide (thick), 400 m long scarp body-remnant of semi-consolidated alluvial fan deposits is exposed along the coast. The touristic village of Cala Gonone has been rapidly expanding in these last few decades over the mid to lower parts of two coalescing alluvial fans and along the coastal marine scarp edge. The village thus became exposed to natural hazards such as sudden overland and creek floods and debris flows typical of alluvial fans, local rock falls, and to instability of the costal scarp due to wave erosion during extreme sea storms. As commonly occurring elsewhere and since antiquity, the risk perception of such events is low because of the centennial, millennial of longer recurrence. Such perception does not negate the hazards but a long event recurrence can be accepted as a reasonable risk for the human’s activity in certain areas. However, serious consideration should be given to potential problems and plan and build for amelioration and defense. Many examples of what can occur and could be done exist from the careful safe-location of the ancient Roman villas to the recent disastrous cases where parts of ill-located villages have been destroyed by debris flows and floods along alluvial fans. The evidence of what did and could still happen in the Cala Gonone and similar other area is clearly imprinted on the landscape (geology, geomorphology, and with relative details in the stratigraphy as sedimentology of the deposits) and driven by climatic conditions and human activities.

Background and Aims Complex modifications of angiosperm flowers often function for precise pollen placement on pollinators and to promote cross-pollination. We explore the functional significance of the unusually elaborate morphology of Gloriosa superba flowers, which are divided into one hermaphrodite meranthium and five male meranthia (functional pollination units of a single flower). Methods We used controlled pollination experiments, floral measurements, pollen load analyses and visitor observations in four populations of G. superba in South Africa to determine the breeding system, mechanism of pollination and role of flower in the promotion of cross-pollination. Key Results We established that G. superba is self-compatible, but reliant on pollinators for seed production. Butterflies, in particular the pierid Eronia cleodora, were the primary pollinators (>90 % of visitors). Butterflies brush against the anthers and stigma during nectar feeding and pollen is carried on their ventral wing surfaces. Butterfly scales were positively correlated with the number of pollen grains on stigmas. We demonstrate that the styles were orientated towards clearings in the vegetation and we confirm that the highest proportion of initial visits was to hermaphrodite meranthia pointing towards clearings. Conclusions The flower morphology of G. superba results in effective pollen transfer on the wings of butterfly visitors. The style-bearing hermaphrodite meranthium of the flowers orientates towards open spaces in the vegetation, thus increasing the probability that butterflies land first on the hermaphrodite meranthium. This novel aspect of flower orientation is interpreted as a mechanism that promotes cross-pollination.

Global database of Orchidaceae to explore the frequency of different breeding systems, means of pollinator attraction, and pollinator specificity, and how these features vary geographically and by growth habit.