Spatial proteomics revealed a CX3CL1-dependent crosstalk between the urothelium and relocated macrophages through IL-6 during an acute bacterial infection in the urinary bladder

The urothelium of the urinary bladder represents the first line of defense. However, uropathogenic E. coli (UPEC) damage the urothelium and cause acute bacterial infection. Here, we demonstrate the crosstalk between macrophages and the urothelium stimulating macrophage migration into the urothelium. Using spatial proteomics by MALDI-MSI and LC-MS/MS, a novel algorithm revealed the spatial activation and migration of macrophages. Analysis of the spatial proteome unravelled the coexpression of Myo9b and F4/80 in the infected urothelium, indicating that macrophages have entered the urothelium upon infection. Immunofluorescence microscopy additionally indicated that intraurothelial macrophages phagocytosed UPEC and eliminated neutrophils. Further analysis of the spatial proteome by MALDI-MSI showed strong expression of IL-6 in the urothelium and local inhibition of this molecule reduced macrophage migration into the urothelium and aggravated the infection. After IL-6 inhibition, the expression of matrix metalloproteinases and chemokines, such as CX3CL1 was reduced in the urothelium. Accordingly, macrophage migration into the urothelium was diminished in the absence of CX3CL1 signaling in Cx3cr1gfp/gfp mice. Conclusively, this study describes the crosstalk between the infected urothelium and macrophages through IL-6-induced CX3CL1 expression. Such crosstalk facilitates the relocation of macrophages into the urothelium and reduces bacterial burden in the urinary bladder. © 2020, The Author(s)

Clinical outcome following acute ischaemic stroke relates to both activation and autoregulatory inhibition of cytokine production

BACKGROUND: As critical mediators of local and systemic inflammatory responses, cytokines are produced in the brain following ischaemic stroke. Some have been detected in the circulation of stroke patients, but their role and source is unclear. Focusing primarily on interleukin(IL)-1-related mechanisms, we serially measured plasma inflammatory markers, and the production of cytokines by whole blood, from 36 patients recruited within 12 h and followed up to 1 year after acute ischaemic stroke (AIS). RESULTS: Admission plasma IL-1 receptor antagonist (IL-1ra) concentration was elevated, relative to age-, sex-, and atherosclerosis-matched controls. IL-1β, soluble IL-1 receptor type II, tumour necrosis factor (TNF)-α, TNF-RII, IL-10 and leptin concentrations did not significantly differ from controls, but peak soluble TNF receptor type I (sTNF-RI) in the first week correlated strongly with computed tomography infarct volume at 5–7 days, mRS and BI at 3 and 12 months. Neopterin was raised in patients at 5–7 d, relative to controls, and in subjects with significant atherosclerosis. Spontaneous IL-1β, TNF-α and IL-6 gene and protein expression by blood cells was minimal, and induction of these cytokines by lipopolysaccharide (LPS) was significantly lower in patients than in controls during the first week. Minimum LPS-induced cytokine production correlated strongly with mRS and BI, and also with plasma cortisol. CONCLUSION: Absence of spontaneous whole blood gene activation or cytokine production suggests that peripheral blood cells are not the source of cytokines measured in plasma after AIS. Increased plasma IL-1ra within 12 h of AIS onset, the relationship between sTNF-RI and stroke severity, and suppressed cytokine induction suggests early activation of endogenous immunosuppressive mechanisms after AIS

Adding 6 months of androgen deprivation therapy to postoperative radiotherapy for prostate cancer: a comparison of short-course versus no androgen deprivation therapy in the RADICALS-HD randomised controlled trial

Background Previous evidence indicates that adjuvant, short-course androgen deprivation therapy (ADT) improves metastasis-free survival when given with primary radiotherapy for intermediate-risk and high-risk localised prostate cancer. However, the value of ADT with postoperative radiotherapy after radical prostatectomy is unclear. Methods RADICALS-HD was an international randomised controlled trial to test the efficacy of ADT used in combination with postoperative radiotherapy for prostate cancer. Key eligibility criteria were indication for radiotherapy after radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to radiotherapy alone (no ADT) or radiotherapy with 6 months of ADT (short-course ADT), using monthly subcutaneous gonadotropin-releasing hormone analogue injections, daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as distant metastasis arising from prostate cancer or death from any cause. Standard survival analysis methods were used, accounting for randomisation stratification factors. The trial had 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 80% to 86% (hazard ratio [HR] 0·67). Analyses followed the intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov, NCT00541047. Findings Between Nov 22, 2007, and June 29, 2015, 1480 patients (median age 66 years [IQR 61–69]) were randomly assigned to receive no ADT (n=737) or short-course ADT (n=743) in addition to postoperative radiotherapy at 121 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 9·0 years (IQR 7·1–10·1), metastasis-free survival events were reported for 268 participants (142 in the no ADT group and 126 in the short-course ADT group; HR 0·886 [95% CI 0·688–1·140], p=0·35). 10-year metastasis-free survival was 79·2% (95% CI 75·4–82·5) in the no ADT group and 80·4% (76·6–83·6) in the short-course ADT group. Toxicity of grade 3 or higher was reported for 121 (17%) of 737 participants in the no ADT group and 100 (14%) of 743 in the short-course ADT group (p=0·15), with no treatment-related deaths. Interpretation Metastatic disease is uncommon following postoperative bed radiotherapy after radical prostatectomy. Adding 6 months of ADT to this radiotherapy did not improve metastasis-free survival compared with no ADT. These findings do not support the use of short-course ADT with postoperative radiotherapy in this patient population

Duration of androgen deprivation therapy with postoperative radiotherapy for prostate cancer: a comparison of long-course versus short-course androgen deprivation therapy in the RADICALS-HD randomised trial

Background Previous evidence supports androgen deprivation therapy (ADT) with primary radiotherapy as initial treatment for intermediate-risk and high-risk localised prostate cancer. However, the use and optimal duration of ADT with postoperative radiotherapy after radical prostatectomy remains uncertain. Methods RADICALS-HD was a randomised controlled trial of ADT duration within the RADICALS protocol. Here, we report on the comparison of short-course versus long-course ADT. Key eligibility criteria were indication for radiotherapy after previous radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to add 6 months of ADT (short-course ADT) or 24 months of ADT (long-course ADT) to radiotherapy, using subcutaneous gonadotrophin-releasing hormone analogue (monthly in the short-course ADT group and 3-monthly in the long-course ADT group), daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as metastasis arising from prostate cancer or death from any cause. The comparison had more than 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 75% to 81% (hazard ratio [HR] 0·72). Standard time-to-event analyses were used. Analyses followed intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov , NCT00541047 . Findings Between Jan 30, 2008, and July 7, 2015, 1523 patients (median age 65 years, IQR 60–69) were randomly assigned to receive short-course ADT (n=761) or long-course ADT (n=762) in addition to postoperative radiotherapy at 138 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 8·9 years (7·0–10·0), 313 metastasis-free survival events were reported overall (174 in the short-course ADT group and 139 in the long-course ADT group; HR 0·773 [95% CI 0·612–0·975]; p=0·029). 10-year metastasis-free survival was 71·9% (95% CI 67·6–75·7) in the short-course ADT group and 78·1% (74·2–81·5) in the long-course ADT group. Toxicity of grade 3 or higher was reported for 105 (14%) of 753 participants in the short-course ADT group and 142 (19%) of 757 participants in the long-course ADT group (p=0·025), with no treatment-related deaths. Interpretation Compared with adding 6 months of ADT, adding 24 months of ADT improved metastasis-free survival in people receiving postoperative radiotherapy. For individuals who can accept the additional duration of adverse effects, long-course ADT should be offered with postoperative radiotherapy. Funding Cancer Research UK, UK Research and Innovation (formerly Medical Research Council), and Canadian Cancer Society

Investigation on Efficacy of Azolla with Different Organic Fertilizers as Plant Growth Enhancer on Chili (Var.Super)

Chili is one of the widely used spices all around the world. Over usage of inorganic fertilizer to satisfy ever growing demand for food leads to detrimental impacts on human health and environment. Therefore, this study was intended to investigate the efficiency of azolla with different organic fertilizer on growth and yield of chili, variety Super. A field experiment was carried out with five different treatments, T1-Azolla+Compost, T2-Azolla+Vermicompost, T3- Azolla+Panchagowvya, T4-Azolla+Vermitea, and T5-Control with five replicates and laid in randomized complete block design. The prepared organic fertilizers were applied from two weeks of transplanting and continued for 12 weeks in one week interval. Then the growth parameter such as mean plant height, number of leaves per plant, leaf width, number of flowers, pod length, pod diameter, pod weight and total yield were measured in different time duration according to the type of measurement. According to the results, T2 showed better performance in terms of mean plant height (45 cm), number of leaves per plant (43 cm), leaf width (2.8 cm), pod length (4.34 cm), individual pod weight (6.94 g), and total yield (176.8 g) whereas comparatively lower performance was observed in control. Considering the significant differences recorded in the statistical analysis for all the parameters other than the mean number of flowers, it can be concluded that the application of Azolla solution with vermicompost improves the growth performance and yield of chili. Therefore, it is recommended to adapt this combination as an organic fertilizer among the farmers to reduce the usage of inorganic fertilizers.   Keywords: Azolla solution, Demand, Growth parameters, Inorganic fertilize

NGC 146: A young open cluster with a Herbig Be star and intermediate mass pre-main sequence stars

We present UBV CCD photometry and low-resolution spectra of stars in the field of the young open cluster NGC 146. UBV photometry of 434 stars were used to estimate the E(B$-$V) reddening of 0.55 $\pm$ 0.04 mag and BV photometry of 976 stars were used to estimate a distance modulus of (m$-$M)$_0$ = 12.7 $\pm$0.2 mag, corresponding to a distance of 3470$^{+335}_{-305}$ pc. We estimated 10 -- 16 Myr as the turn-off age for the upper main sequence of the cluster using isochrones and synthetic colour magnitude diagrams. We identified two B type stars with H$_\alpha$ in emission and located on the MS using slit-less spectra. A higher resolution spectrum of the brighter Be star indicated the presence of a number of emission lines, with some lines showing the signature of gas infall. This star was found to be located in the region of Herbig Ae/Be stars in the (J$-$H) vs (H$-$K) colour-colour diagram. Thus, we identify this star as a Herbig Be star. On the other hand, 54 stars were found to show near infrared excess, of which 17 were found to be located in the region of Herbig Ae/Be stars and 18 stars were found to be located in the region of Be stars in the NIR colour-colour diagram. Thus NGC 146 is a young cluster with a large number of intermediate mass pre-main sequence stars. The turn-on age of the cluster is found to be $\sim$ 3 Myr. Though NGC 146 shows an older turn off, the bulk of stars in this cluster seems to belong to the younger population of 3 Myr

Detection, Identification, and Antimicrobial Susceptibility of Campylobacter spp. and Salmonella spp. from Free-ranging Nonhuman Primates in Sri Lanka

Infections with Campylobacter spp. and Salmonella spp. are the most frequently reported causes of human bacterial enteritis. Warm-blooded animals, including livestock, pets, and wildlife, can be carriers of the bacteria and may contaminate the environment and food products. The present study investigated the occurrence of Campylobacter spp. and Salmonella spp. in fecal pat samples from free-ranging toque macaques (Macaca sinica) and tufted gray langurs (Semnopithecus priam) collected in March-May 2015 in Sri Lanka. In 58 samples from toque macaques, Campylobacter jejuni was isolated in 10 (17%), Campylobacter coli in four (7%), and Salmonella enterica subsp. enterica serovar Virchow in two (3%). None of the bacteria were isolated in the 40 samples from tufted gray langurs. Pulse-field gel electrophoresis and multilocus sequence typing identified six profiles and four clonal complexes of C. jejuni. The isolated Campylobacter spp. showed varying susceptibility to antimicrobial substances. All Campylobacter spp. isolates were susceptible to chloramphenicol, erythromycin, florfenicol, gentamicin, and streptomycin. Four of the C. jejuni were resistant to at least one of the following: ampicillin, ciprofloxacin, nalidixic acid, and tetracycline, and one of the isolates was multidrug resistant. All four C. coli were resistant to ampicillin, whereas the two Salmonella Virchow strains were susceptible to all antibiotics tested. The presence of Campylobacter spp. and Salmonella spp. in toque macaques may have an impact on the conservation of endangered primates and public health in Sri Lanka