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The present study aimed to investigate the effect of encapsulated Cosmos caudatus leaf extract on the physiological conditions, immune competency, and antioxidative status of broiler chickens raised at a high stocking density. After 15 days of rearing, 370 Lohmann broiler chicks were assigned to five treatment groups, including T0 (chicks were raised at a density of 10 birds/m2 and received no additive), T1 (chicks were raised at a density of 16 birds/m2 and received no additive), T2 (chicks were raised at a density of 16 birds/m2 and received 0.5 g/kg encapsulated C. caudatus leaf extract), T3 (chicks were raised at a density of 16 birds/m2 and received 1.0 g/kg additive), and T4 (chicks were raised at a density of 16 birds/m2 and received 1.5 g/kg additive). On days 28 and 42, blood samples from two chicks per pen were collected. On day 42, the chicks that had been blood-sampled were sacrificed, and blood samples and lymphoid organs (i.e., bursa of Fabricius, spleen, and thymus) were collected. The daily weight gain and feed efficiency of broilers (PC. caudatus leaf extract, especially at a rate of 1.5 g/kg, could alleviate the cortical and lymphocyte cell depletion of broilers. Regardless of the stocking density effect, dietary supplementation with encapsulated C. caudatus leaf extract at doses of 0.5 and 1.0 g/kg could improve the daily weight gain of broilers.

Background and Aim: In the post-antibiotic era, consumer demand for healthy and safe meats has prompted poultry producers to seek alternative effective feed additives. This study aimed to investigate the effects of a novel natural feed additive based on a mixture of Averrhoa bilimbi L. fruit filtrate, wheat bran, and Saccharomyces cerevisiae on the growth rate, internal organ weight, and breast meat characteristics of broilers. Materials and Methods: A total of 280 1-day-old chicks were divided into one control (CNTRL; feed without additives) and three treatment groups: NOV25, feed with 2.5 g/kg novel additive; NOV50, feed with 5.0 g/kg novel additive; and NOV100, feed with 10 g/kg novel additive. The body weight (BW), feed intake (FI), and feed conversion ratio (FCR) were measured weekly. On day 35, the chickens from each group were slaughtered, and their internal organs and breast meat samples were collected. Results: The BW of broilers in NOV100 was greater (p=0.016) than that in the other groups. The FCRs in the treatments groups were lower (p

Several scoring systems predict risks of clinically relevant postoperative pancreatic fistula after pancreatectomy, but none have emerged as the gold standard. This study aimed to evaluate the accuracy of postoperative day 1 drain amylase and serum C-reactive protein levels in predicting clinically relevant postoperative pancreatic fistula compared with intraoperative pancreatic characteristics.Patients who underwent pancreatectomy between 2017 and 2019 were included prospectively. Cutoff values were determined using receiver operating characteristic curves, and a score combining postoperative day 1 drain amylase and serum C-reactive protein was tested in a multivariate logistic regression model to evaluate clinically relevant postoperative pancreatic fistula risk.A total of 274 pancreatic resections (182 pancreaticoduodenectomies and 92 distal pancreatectomies) were included. The pancreatic gland texture was "soft" in 47.8% (n = 131), and 55.8% (n = 153) had a small size main pancreatic duct (≤3 mm). Clinically relevant postoperative pancreatic fistula occurred in 58 patients (21.2%). Drain amylase ≥1,000 UI/L and serum C-reactive protein ≥90 mg/L were identified as the optimal cutoffs to predict clinically relevant postoperative pancreatic fistula. On multivariate analysis these cutoffs were independent predictors of clinically relevant postoperative pancreatic fistula after both pancreaticoduodenectomies (drain amylase: P.001, serum C-reactive protein: P = .006) and distal pancreatectomies (drain amylase: P = .009, serum C-reactive protein: P = .001). The postoperative day 1 "90-1000" model, a 2-value score relying on these cutoffs, significantly (P.001) outperformed intraoperative pancreatic parenchymal characteristics in predicting clinically relevant postoperative pancreatic fistula after both pancreaticoduodenectomies and distal pancreatectomies. A postoperative day 1 "90-1000" score = 0 had a negative predictive value of 97% and 94%, respectively, after pancreaticoduodenectomy and distal pancreatectomies.A combined score relying on postoperative day 1 values of drain amylase and serum C-reactive protein levels was accurate in predicting risks of clinically relevant postoperative pancreatic fistula after pancreatectomy.

BACKGROUND Tomato is widely consumed throughout the world for its flavor and nutritional value. This functional food largely depends on the implementation of new strategies to maintain the nutraceutical value, e.g. lycopene concentration, and overcome the challenges of sustainable production and food security. The use of arbuscular mycorrhizal fungi (AMF)-based biostimulants represents one of the most promising tools for sustainable management of agricultural soils, being fundamental for organic food production, reducing fertilizers and pesticides use, and decreasing environmental damage. This study aimed at elucidating whether native arbuscular mycorrhizal fungi (AMF) could positively affect tomato yield and lycopene concentration. RESULTS Native AMF inoculum consisted of two inoculum types: the single species Claroideoglomus claroideum, and a mix of Scutellospora calospora, Acaulospora laevis, Claroideoglomus claroideum, and Claroideoglomus etunicatum. At the end of the study up to 78% of the root system was colonized by single inoculum. Tomato diameters in single and mix mycorrhizal plants showed increases of 80% and 35% respectively. Fresh weights were 84% and 38% higher with single and mix inocula compared with the controls, respectively. The lycopene concentration in tomato fruits of plants with single and mix inoculum was higher than controls. The lycopene concentration was 124.5% and 113.9% greater in single and mix than non-inoculated plants. CONCLUSION Tomato diameters, fresh weight and lycopene concentration was significantly higher in plants colonized by AMF compared with uninoculated plants. Results suggest that the role of single species Claroideoglomus claroideum could generate better plant performance due to its high production of extraradical mycelium. © 2021 Society of Chemical Industry.

Arbuscular mycorrhizal fungi (AMF) represent an important group of root symbionts, given the key role they play in the enhancement of plant nutrition, health, and product quality. The services provided by AMF often are facilitated by large and diverse beneficial bacterial communities, closely associated with spores, sporocarps, and extraradical mycelium, showing different functional activities, such as N2 fixation, nutrient mobilization, and plant hormone, antibiotic, and siderophore production and also mycorrhizal establishment promotion, leading to the enhancement of host plant performance. The potential functional complementarity of AMF and associated microbiota poses a key question as to whether members of AMF-associated bacterial communities can colonize the root system after establishment of mycorrhizas, thereby becoming endophytic. Root endophytic bacterial communities are currently studied for the benefits provided to host plants in the form of growth promotion, stress reduction, inhibition of plant pathogens, and plant hormone release. Their quantitative and qualitative composition is influenced by many factors, such as geographical location, soil type, host genotype, and cultivation practices. Recent data suggest that an additional factor affecting bacterial endophyte recruitment could be AMF and their associated bacteria, even though the mechanisms allowing members of AMF-associated bacterial communities to actually establish in the root system, becoming endophytic, remain to be determined. Given the diverse plant growth–promoting properties shown by AMF-associated bacteria, further studies are needed to understand whether AMF may represent suitable tools to introduce beneficial root endophytes in sustainable and organic agriculture where the functioning of such multipartite association may be crucial for crop production.

COVID-19 is associated with high mortality in patients with haematological malignancies (HM) and rate of seroconversion is unknown. The ITA-HEMA-COV project (NCT04352556) investigated patterns of seroconversion for SARS-CoV-2 IgG in patients with HMs. A total of 237 patients, SARS-CoV-2 PCR-positive with at least one SARS-CoV-2 IgG test performed during their care, entered the analysis. Among these, 62 (26·2%) had myeloid, 121 (51·1%) lymphoid and 54 (22·8%) plasma cell neoplasms. Overall, 69% of patients (164 of 237) had detectable IgG SARS-CoV-2 serum antibodies. Serologically negative patients (31%, 73 of 237) were evenly distributed across patients with myeloid, lymphoid and plasma cell neoplasms. In the multivariable logistic regression, chemoimmunotherapy [odds ratio (OR), 3·42; 95% confidence interval (CI), 1·04–11·21; P=0·04] was associated with a lower rate of seroconversion. This effect did not decline after 180days from treatment withdrawal (OR, 0·35; 95% CI: 0·11–1·13; P=0·08). This study demonstrates a low rate of seroconversion in HM patients and indicates that treatment-mediated immune dysfunction is the main driver. As a consequence, we expect a low rate of seroconversion after vaccination and thus we suggest testing the efficacy of seroconversion in HM patients.

The present experiment investigated the impact of administrations of lactic fermented turmeric, black pepper or a mixture of both into diets on the hematological parameters and intestinal ecology and morphology of broilers. Three hundred and ninety two broilers were spread to T0 (control), T1 (feed administrated with 1% lactic fermented turmeric), T2 (feed administrated with 1% lactic fermented black pepper) and T3 (feed administrated with 1% lactic fermented turmeric and 1% lactic fermented black pepper). Blood sampling was conducted at days 21 and 35, while intestinal sampling was at day 35. At day 21, erythrocyte counts were greater (P ≤ 0.05) for T1 compared to that for T2 and T3. Broilers in T1 had deceased (P ≤ 0.05) serum total triglyceride compared to that of T0 and T3. The concentration of globulin was reduced (P ≤ 0.05) in T3 than that in T0 and T1. At day 35, serum triglyceride level was more elevated (P ≤ 0.05) in T0 compared to that in T2 and T3 groups. Serum cholesterol as well as low-density lipoprotein levels were lower (P ≤ 0.05) in T3 than that in T0. Compared to other broilers, total protein and globulin concentrations were lower (P ≤ 0.05) in T3. A bigger (P ≤ 0.05) ratio of albumin to globulin (A/G) was seen in T3 compared to that in T1 and T2. Creatinine was greater (P ≤ 0.05) in T1 than in others. The titer of antibody toward vaccine of Newcastle disease was elevated (P ≤ 0.05) in T1 and T2 than in T0. The LAB counts were enhanced (P ≤ 0.05) in ileum of T1 and T2 than that in T0. Compared to T0, the villi height of jejunum was elevated (P ≤ 0.05) in treated broilers. In conclusion, administration of fermented turmeric powder improved erythrocyte counts, antibody titer toward Newcastle disease vaccine, LAB population and jejunal villi height of broilers.

Photodynamic therapy (PDT) represents a promising therapeutic modality for cancer. Here we used an orthogonal nanoarchitectonics approach (genetic/chemical) to engineer M13 bacteriophages as targeted vectors for efficient photodynamic killing of cancer cells. M13 was genetically refactored to display on the phage tip a peptide (SYPIPDT) able to bind the epidermal growth factor receptor (EGFR). The refactored M13(EGFR) phages demonstrated EGFR-targeted tropism and were internalized by A431 cancer cells, that overexpress EGFR. Using an orthogonal approach to the genetic display, M13(EGFR) phages were then chemically modified, conjugating hundreds of Rose Bengal (RB) photosensitizing molecules on the capsid surface, without affecting the selective recognition of the SYPIPDT peptides. Upon internalization, the M13(EGFR)-RB derivatives generated intracellularly reactive oxygen species, activated by an ultralow intensity white light irradiation. The killing activity of cancer cells is observed at picomolar concentrations of the M13(EGFR) phage.

Breast Cancer (BC) is a leading cause of death in women, currently affecting 13% of female population worldwide. First-line clinical treatments against Estrogen Receptor positive (ER+) BC rely on suppressing estrogen production, by inhibiting the aromatase (AR) enzyme, or on blocking estrogen-dependent pro-oncogenic signaling, by targeting Estrogen Receptor (ER) α with selective Modulators/Degraders (SERMs/SERDs). The development of dual acting molecules targeting AR and ERα represents a tantalizing alternative strategy to fight ER + BC, reducing the incidence of adverse effects and resistance onset that limit the effectiveness of these gold-standard therapies. Here, in silico design, synthesis, biological evaluation and an atomic-level characterization of the binding and inhibition mechanism of twelve structurally related drug-candidates enable the discovery of multiple compounds active on both AR and ERα in the sub-μM range. The best drug-candidate 3a displayed a balanced low-nanomolar IC50 towards the two targets, SERM activity and moderate selectivity towards a BC cell line. Moreover, most of the studied compounds reduced ERα levels, suggesting a potential SERD activity. This study dissects the key structural traits needed to obtain optimal dual acting drug-candidates, showing that multitarget compounds may be a viable therapeutic option to counteract ER + BC.

Glioblastoma (GBM) is one of the most malignant and devastating brain tumors. The presence of highly therapy-resistant GBM cell subpopulations within the tumor mass, rapid invasion into brain tissues and reciprocal interactions with stromal cells in the tumor microenvironment contributes to an inevitable fatal prognosis for the patients. We highlight the most recent evidence of GBM cell crosstalk with mesenchymal stem cells (MSCs), which occurs either by direct cell-cell interactions via gap junctions and microtubules or cell fusion. MSCs and GBM paracrine interactions are commonly observed and involve cytokine signaling, regulating MSC tropism toward GBM, their intra-tumoral distribution, and immune system responses. MSC-promoted effects depending on their cytokine and receptor expression patterns are considered critical for GBM progression. MSC origin, tumor heterogeneity and plasticity may also determine the outcome of such interactions. Kinins and kinin-B1 and -B2 receptors play important roles in information flow between MSCs and GBM cells. Kinin-B1 receptor activity favors tumor migration and fusion of MSCs and GBM cells. Flow and image (tissue) cytometry are powerful tools to investigate GBM cell and MSC crosstalk and are applied to analyze and characterize several other cancer types.

Proteasome inhibition represents an important anticancer strategy. Here, we studied the mechanisms at the basis of the pro-apoptotic activity of the standardized decoction of Hemidesmus indicus, a plant evoking a complex anticancer activity, and explored its inhibition of proteasome activity in human leukemia cells. Additionally, we preliminary tested the cytotoxicity of some H. indicus’s phytochemicals on leukemia cells and their intestinal absorption on a human intestinal epithelium model consisting of a monolayer of differentiated Caco2 cells. We observed a potent antileukemic effect for H. indicus, imputable to the modulation of different critical targets at protein and mRNA levels and the reduction of the 26S proteasome expression. We found that some phytomarkers of H. indicus decoction passed through the enterocyte monolayer. Overall, our study supports the pharmacological potential of H. indicus, which can represent an interesting botanical drug in the oncological area.

The work investigated the impact of a fermented mixture of papaya leaf and seed meal (FERM) on production traits, biochemical indices, intestinal ecology and carcass proportion of the Indonesian indigenous crossbred chickens (ICC). A 300 day-old ICC were assigned to five groups and fed on starter (1–4 weeks) and finisher diets (5–8 weeks) containing corn, soybean (CONT) or diets containing 1% (FERM1), 2.5% (FERM25), 5% (FERM5) or 7.5% (FERM75) of FERM. On week 8, blood was collected from birds prior to slaughter. The increased levels of FERM linearly reduced (P < 0.05) feed conversion and feed cost per kg weight gain, and increased (P < 0.05) income over feed cost of chickens. The relative weight of proventriculus linearly reduced (P < 0.05) with the enhanced contents of dietary FERM. The haemoglobin, erythrocytes and haematocrit values linearly reduced (P < 0.05) as the proportions of FERM enhanced. The mean corpuscular hemoglobin (MCH) levels were greater (P < 0.05) in FERM1 and FERM25 than that in control. The number of heterophils was greater (P < 0.05) in FERM1 than that in FERM25 and FERM5, but was not distinct from CONT and FERM75. The increased levels of FERM linearly increased (P < 0.05) thrombocytes values. Feeding FERM at 2.5% resulted in higher (P < 0.05) high-density lipoprotein (HDL) to low-density lipoprotein (LDL) ratio than that of control and FERM1. FERM reduced (P < 0.05) serum content of alanine aminotransferase (ALT). FERM75 had higher (P < 0.05) level of creatinine than control, FERM1 and FERM25. In duodenum, the increased levels of FERM linearly (P < 0.05) increased crypt depth (CD). At 7.5%, FERM reduced (P < 0.05) CD and enhanced (P < 0.05) villus height (VH) to CD ratio in jejunum. In ileum, CD linearly elevated (P < 0.05) with the increased FERM, while feeding 2.5% FERM enhanced (P < 0.05) VH and VH to CD ratio. In conclusion, FERM was beneficial in improving economic performance, immune responses, physiological condition and intestinal morphology of ICC.

The study aimed to evaluate effect of the used rice fermented using Chrysonillia crassa as a functional feed on biochemical and haematological parameters , microbial populations, internal organ weight and performance of broilers. The study used o ne hundred and fifty unsexed one-day-old chicks with average body weight of 48.30 ± 4.13g. At day 1 to 7, the chicks were given commercial feed. On day 8 onward , the chickens were randomized distributed to one of three experimental diets of 50 chicks each. The chicks were given formulated diet containing fermented used rice based-functional feed (FF). Drinking water was provided ad libitum . The study used completely randomized design with three experimental diets : T0 (diet without FF), T1 (diet contai ned 5% FF), and T2 (diet contained 10% FF). The chicken fed the used rice fermented increased in cholesterol , high density lipoprotein status, mean corpuscular haemoglobin concentration, lactic acid bacteria, heart weight. and decreased in mean corpuscular volume , E.coli , jejunum weight , but no affect on body weight gain, feed convertion ratio and feed comsumption. In conclusion that the fermented used rice improved the physiological conditions ,intestinal bacterial population and had no detrimental effect on performance of broiler .

The study investigated gut ecology and morphology of the Indonesian indigenous crossbred chickens (IICC) supplemented with the combination of formic acid and Saccharomyces cerevisiae. Two hundreds day-old IICC were distributed to T0 (control diet), T1 (T0 + 0.2% formic acid), T2 (T0 + 0.3% S. cerevisiae), T3 (T0 + 0.2% formic acid and 0.3% S. cerevisiae). Excreta was collected at week 8, while intestinal ecology and morphology were determined at week 9. In duodenum, T3 chicks showed higher and wider (P0.05). IIn conclusion, the inclusion of formic acid and S. cerevisiae in diets improved intestinal ecology and morphology. The IICC chickens fed with formic acid and S. cerevisiae exhibited improved growth performance and nutrient digestibility.

The study investigated the influence of dietary incorporation of formic acid, butyric acid or their blends on growth rate, haematological indices and intestinal morphometric of broilers. A number of 240 Lohmann MB-202 chicks were pass rounded to four dietary groups, including CONT (chicks taking in basal feed with no additive), BTRT (chicks receiving basal feed with 0.03% butyric acid), FRMT (chicks receiving basal feed with 0.1% formic acid) and BTRT+FRMT (chicks receiving basal feed containing 0.03% butyric acid and 0.1% formic acid). Weight of chicks and intake were measured every week, while blood sample was collected at day 21 and 35. At day 35, birds were slaughtered and small intestinal segments and digesta were collected. At day 21, body weight was bigger (P

Aim: The study investigated the effect of feeding fermented mixture of cassava pulp and Moringa oleifera leaf meal (FCPMO) on the immune responses, antioxidative status, biochemical parameters, and intestinal ecology of broiler chickens. Materials and Methods: Four hundred Lohmann broiler chickens were distributed to four groups of diets including CONT (corn-soybean-based feed with no additive), BACI (corn-soybean-based diet supplemented with 0.1% zinc bacitracin), FERM (diet containing 20% FCPMO), and FERB (diet containing 20% FCPMO and added with 0.1% Bacillus subtilis). At days 4, 14, and 19, the chicks were vaccinated using commercial Newcastle disease-infectious bursal disease (ND-IBD), IBD, and ND vaccines, respectively. At day 35, blood was sampled and digesta was obtained from the ileum and caecum. Furthermore, the duodenal segment was obtained. Results: The BACI, FERM, and FERB groups had higher (p0.05) on cecal volatile fatty acids production. Conclusion: Feeding FCPMO improved immune responses, antioxidative status, and physiological conditions, but had less effect on the intestinal ecology of broilers.

The incidence of endometrial cancer (EC) is increasing in developed countries. The most frequent is the endometrioid subtype with usually good prognosis; nevertheless, some cases escape this paradigm and may have recurrence. A recent study from The Cancer Genome Atlas suggested to implement the EC analysis by molecular profile for improving diagnosis, prognosis, and therapeutic treatment. The present preliminary study was performed on 15 G3 endometrioid endometrial cancers (G3 EEC) for the identification of somatic mutations in a panel of specific exons in selected genes as ARID1A, CTNNB1, KRAS, PIK3CA, POLE, PTEN, and TP53. The combined procedure, based on the Sanger sequencing and PCR-high-resolution melting analysis, allowed the identification of variations of the selected gene panel in most of patients (93%) of our cohort. The overall evaluation of mutational load exhibited that the most frequent mutated genes were PTEN (93%), followed by PIK3CA (47%) suggesting a deep involvement of PI3K pathway alteration in G3 EEC. Mutations in TP53 (27%), ARID1A (27%), POLE (13%), and at the lower level in KRAS and CTNNB1 (7%) were also observed (exclusively in FIGO III stage patients). The evaluation of the mutations of our proposed panel (ARID1A, CTNNB1, KRAS, PIK3CA, POLE, PTEN, TP53) is suitable to improve the characterization of G3 EEC and could suggest targetable pathways for development of personalized treatments.

International audience; Safe and effective vaccines against coronavirus disease 2019 (COVID-19) are essential for ending the ongoing pandemic. Although impressive progress has been made with several COVID-19 vaccines already approved, it is clear that those developed so far cannot meet the global vaccine demand alone. We describe a COVID-19 vaccine based on a replication-defective gorilla adenovirus expressing the stabilized prefusion severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein named GRAd-COV2. We assessed the safety and immunogenicity of a single-dose regimen of this vaccine in healthy younger and older adults to select the appropriate dose for each age group. For this purpose, a phase 1, dose-escalation, open-labeled trial was conducted including 90 healthy participants (45 aged 18 to 55 years old and 45 aged 65 to 85 years old) who received a single intramuscular administration of GRAd-COV2 at three escalating doses. Local and systemic adverse reactions were mostly mild or moderate and of short duration, and no serious adverse events were reported. Four weeks after vaccination, seroconversion to spike protein and receptor binding domain was achieved in 43 of 44 young volunteers and in 45 of 45 older participants. Consistently, neutralizing antibodies were detected in 42 of 44 younger-age and 45 of 45 older-age volunteers. In addition, GRAd-COV2 induced a robust and T helper 1 cell (T H 1)-skewed T cell response against the spike protein in 89 of 90 participants from both age groups. Overall, the safety and immunogenicity data from the phase 1 trial support the further development of this vaccine.

Simple Summary Inborn Errors of Immunity (IEI) are a heterogeneous group of disorders characterized by a defect in the function of at least one, and often more, components of the immune system. The overall risk for cancer in children with IEI ranges from 4 to 25%. Several factors, namely, age of the patient, viral infection status and IEI type can influence the development of different cancer types. Immunologists and oncologists should interact to monitor and promptly diagnose the potential development of cancer in known IEI patients, as well as an underlying IEI in newly diagnosed cancers with suggestive medical history or high rate of therapy-related toxicity. The creation of an international registry of IEI cases with detailed information on the occurrence of cancer is fundamental to optimizing the diagnostic process and to evaluating the outcomes of new therapeutic options, with the aim of improving prognosis and reducing comorbidities. Abstract Inborn Errors of Immunity (IEI) are a heterogeneous group of disorders characterized by a defect in the function of at least one, and often more, components of the immune system. The aim of this narrative review is to discuss the epidemiology, the pathogenesis and the correct management of tumours in patients with IEI. PubMed was used to search for all of the studies published over the last 20 years using the keywords: “inborn errors of immunity” or “primary immunodeficiency” and “cancer” or “tumour” or “malignancy”. Literature analysis showed that the overall risk for cancer in children with IEI ranges from 4 to 25%. Several factors, namely, age of the patient, viral infection status and IEI type can influence the development of different cancer types. The knowledge of a specific tumour risk in the presence of IEI highlights the importance of a synergistic effort by immunologists and oncologists in tracking down the potential development of cancer in known IEI patients, as well as an underlying IEI in patients with newly diagnosed cancers. In the current genomic era, the creation of an international registry of IEI cases integrated with malignancies occurrence information is fundamental to optimizing the diagnostic process and to evaluating the outcomes of new therapeutic options, with the hope to obtain a better prognosis for these patients.

Background The benefit of precision medicine based on relatively limited gene sets and often-archived samples remains unproven. PERMED-01 (NCT02342158) was a prospective monocentric clinical trial assessing, in adults with advanced solid cancer, the feasibility and impact of extensive molecular profiling applied to newly biopsied tumor sample and based on targeted NGS (t-NGS) of the largest gene panel to date and whole-genome array-comparative genomic hybridization (aCGH) with assessment of single-gene alterations and clinically relevant genomic scores. Methods Eligible patients with refractory cancer had one tumor lesion accessible to biopsy. Extracted tumor DNA was profiled by t-NGS and aCGH. We assessed alterations of 802 “candidate cancer” genes and global genomic scores, such as homologous recombination deficiency (HRD) score and tumor mutational burden. The primary endpoint was the number of patients with actionable genetic alterations (AGAs). Secondary endpoints herein reported included a description of patients with AGA who received a “matched therapy” and their clinical outcome, and a comparison of AGA identification with t-NGS and aCGH versus whole-exome sequencing (WES). Results Between November 2014 and September 2019, we enrolled 550 patients heavily pretreated. An exploitable complete molecular profile was obtained in 441/550 patients (80%). At least one AGA, defined in real time by our molecular tumor board, was found in 393/550 patients (71%, two-sided 90%CI 68–75%). Only 94/550 patients (17%, 95%CI 14–21) received an “AGA-matched therapy” on progression. The most frequent AGAs leading to “matched therapy” included PIK3CA mutations, KRAS mutations/amplifications, PTEN deletions/mutations, ERBB2 amplifications/mutations, and BRCA1/2 mutations. Such “matched therapy” improved by at least 1.3-fold the progression-free survival on matched therapy (PFS2) compared to PFS on prior therapy (PFS1) in 36% of cases, representing 6% of the enrolled patients. Within patients with AGA treated on progression, the use of “matched therapy” was the sole variable associated with an improved PFS2/PFS1 ratio. Objective responses were observed in 19% of patients treated with “matched therapy,” and 6-month overall survival (OS) was 62% (95%CI 52–73). In a subset of 112 metastatic breast cancers, WES did not provide benefit in term of AGA identification when compared with t-NGS/aCGH. Conclusions Extensive molecular profiling of a newly biopsied tumor sample identified AGA in most of cases, leading to delivery of a “matched therapy” in 17% of screened patients, of which 36% derived clinical benefit. WES did not seem to improve these results. Trial registration ID-RCB identifier: 2014-A00966-41; ClinicalTrials.gov identifier: NCT02342158.

Lysobacter niastensis belongs to a group of bacterial predators that produce a number of bioactive small molecules endowed with lytic properties toward other microorganisms. Here, we report the draft genome sequence of the type strain DSM 18481 and the identification of gene clusters implicated in the biosynthesis of secondary metabolites.

Background: Natural products are characterized by a complex chemical composition and are capable of concurrently modulate several signalling pathways. Considering the biological complexity of carcinogenesis, natural products represent key components of the therapeutic armamentarium for oncological diseases. The bark of Terminalia arjuna is used in traditional Ayurvedic medicine for its astringent, expectorant, cardiotonic, styptic, and antidysenteric properties. Alongside its traditional uses, Terminalia arjuna exhibits different biological activities including antimutagenic and anticarcinogenic. Objective: This study was designed to evaluate the toxic effects of an alcoholic extract obtained from the bark of T. arjuna on a human T-lymphoblastic cell line (Jurkat). We explored the phytochemical composition and investigated the cytotoxic, cytostatic, genotoxic, and anti-genotoxic effects. Methods: The phytochemical composition was analyzed using spectrophotometric methods; all the biological endpoints were assessed through flow cytometry. Results: The phytochemical screening showed that polyphenols represent about 64% of the extract. Moreover, the extract was cytotoxic on Jurkat cells by inducing both apoptosis and necrosis, and blocked the cell cycle in the G2/M phase. Additionally, it was found that the extract lacks any genotoxic effect, but was not effective in protecting Jurkat cells from the DNA damage induced by H2O2 and etoposide. Conclusion: The results of our study show the toxic effects of Terminalia arjuna on Jurkat cells and confirm the pivotal role played by natural compounds in the oncological field. Further studies should be performed to better understand its clinical potential and deepen its toxicological profile.

The study investigated how sprouted-papaya seed meal (SPSM) a total of affected the growth and carcase traits of broilers. Based on a completely randomized design, 390 day-old Lohmann broiler chicks were allotted to CONT (chicks provided with control diet), SEED25 (diet containing 2.5% papaya seed meal), GERM1 (diet containing 1% SPSM), GERM25 (diet containing 2.5% SPSM) and GERM5 (diet containing 5% SPSM). Body weight and feed intake of chickens were weekly recorded from 14 to 36 days of age, while birds (six birds per treatment group; 30 birds in total) were slaughtered at day 36. Feeding SPSM up to 5% did not impair (p > 0.05) broilers’ growth or feed intake. At 5%, SPSM compromised (p 0.05) detrimental effects, while papaya seed meal at the same proportion lowered (p

Aminobacter carboxidus is a soil Gram-negative alphaproteobacterium belonging to the physiological group of carboxydobacteria which aerobically oxidize CO to CO2. Here, we report the draft genome sequence of the A. carboxidus DSM 1086 type strain and the identification of both form I and form II CO dehydrogenase systems in this strain.

We present the development of a custom-made two-photon light-sheet microscope optimized for high-speed (5 Hz) volumetric imaging of zebrafish larval brain for the analysis of neuronal physiological and pathological activity. High-speed volumetric two-photon light-sheet microscopy is challenging to achieve, due to constrains on the signal-to-noise ratio. To maximize this parameter, we optimized our setup for high peak power of excitation light, while finely controlling its polarization, and we implemented remote scanning of the focal plane to record without disturbing the sample. Two-photon illumination is advantageous for zebrafish larva studies since infra-red excitation does not induce a visual response, that otherwise would affect the neuronal activity. In particular, we were able to record whole-brain neuronal activity of the larva with high temporal- and spatial-resolution during the nocturnal period without affecting the circadian rhythm. Analyzing the spatially resolved power spectra of GCaMP signal, we found significant differences for several frequency bands between the day/night phases in various brain regions. Moreover, we studied the fast dynamics that characterize the acutely induced pathological epileptic activity of the larvae, identifying the brain structures that are more susceptible to the action of the epileptogenic drug. In conclusion, the high speed two-photon light-sheet microscope that we developed is proving to be an important tool to study both the physiological and the pathological activity of the zebrafish larval brain without undesired visual stimulation.

Objective. Ultrasounds (US) use in neural engineering is so far mainly limited to ablation through high intensity focused ultrasound, but interesting preliminary results show that low intensity low frequency ultrasound could be used instead to modulate neural activity. However, the extent of this modulatory ability of US is still unclear, as in in vivo studies it is hard to disentangle the contribution to neural responses of direct activation of the neuron by US stimulation and indirect activation due either to sensory response to mechanical stimulation associated to US, or to propagation of activity from neighboring areas. Here, we aim to show how to separate the three effects and assess the presence of direct response to US stimulation in zebrafish. Approach. We observed in zebrafish larvae brain-wide US-induced activity patterns through calcium imaging microscopy. Sensory response to mechanical stimulation was assessed with a US shield. Activity propagation was assessed with inter-area latency evaluation. Main results. We prove that in selected brain regions the zebrafish’s neural response is mainly due to direct activation, later spreading to the other regions. Shielding the neurons from direct US stimulation resulted in a significantly attenuated response, showing that sensory stimulation does not play a prominent role. Significance. US non-invasive neuromodulatory approach might lead to novel ways to test and control neural activity, and hence to novel neuromodulatory therapies. Future studies will focus on the biophysical structure of directly responsive neurons to capture the mechanisms of US induced activity.

We have reported that short-term stimulation of primary human monocyte-derived macrophages (MDM) with interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α), i.e. M1 polarization, leads to a significant containment of virus replication. Here we show that M1-MDM restimulation with these cytokines 7 days after infection (M12 MDM) promoted an increased restriction of HIV-1 replication characterized by very low levels of virus production near to undetectable levels. In comparison to control and M1-MDM that were not restimulated, M12 MDM showed a stronger reduction of both total and integrated HIV DNA as well as of viral mRNA expression. M12 MDM were characterized by an upregulated expression of restriction factors acting at the level of reverse transcription (RT), including apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3A (APOBEC3A) and APOBEC3G, but not SAM domain and HD domain-containing protein 1 (SAMHD1). M12 MDM also showed an increased expression of Class II Transactivator (CIITA) and Tripartite Motif22 (TRIM22), two negative regulators of proviral transcription, whereas expression and phosphorylation of transcriptional inducers of HIV-1, such as nuclear factor kB (NF-kB) and signal transducer and activator of transcription 1 (STAT1), were not impaired in these cells. The almost quiescent state of the infection in M12 MDM was promptly reversed by coculture with mitogen-stimulated leukocytes or cell incubation with their filtered culture supernatant. M12 MDM harbored replication-competent HIV-1 as virus spreading following cell stimulation was fully prevented by the RT inhibitor lamivudine/3TC. Selective reactivation of proviral expression in M12 MDM, but not in control or in M1-MDM that were not restimulated, was confirmed in cells infected with single round Vesicular Stomatitis Virus-G-pseudotyped HIV-1. Thus, M12 MDM represent an in vitro model of reversible, almost quiescent HIV-1 infection of primary human macrophages that could be further exploited for “Cure” related investigations.

To egress from its erythrocyte host, the malaria parasite, Plasmodium falciparum, must destabilize the erythrocyte membrane by activating an erythrocyte tyrosine kinase. Because imatinib inhibits erythrocyte tyrosine kinases and because imatinib has a good safety profile, we elected to determine whether coadministration of imatinib with standard of care (SOC) might be both well tolerated and therapeutically efficacious in malaria patients. Patients with uncomplicated P. falciparum malaria from a region in Vietnam where one third of patients experience delayed parasite clearance (DPC; continued parasitemia after 3 d of therapy) were treated for 3 d with either the region’s SOC (40 mg dihydroartemisinin + 320 mg piperaquine/d) or imatinib (400 mg/d) + SOC. Imatinib + SOC–treated participants exhibited no increase in number or severity of adverse events, a significantly accelerated decline in parasite density and pyrexia, and no DPC. Surprisingly, these improvements were most pronounced in patients with the highest parasite density, where serious complications and death are most frequent. Imatinib therefore appears to improve SOC therapy, with no obvious drug-related toxicities.

Bacteria belonging to the genus Aminobacter are metabolically versatile organisms thriving in both natural and anthropized terrestrial environments. To date, the taxonomy of this genus is poorly defined due to the unavailability of the genomic sequence of A. anthyllidis LMG 26462T and the presence of unclassified Aminobacter strains. Here, we determined the genome sequence of A. anthyllidis LMG 26462T and performed phylogenomic, average nucleotide identity and digital DNA-DNA hybridization analyses of 17 members of genus Aminobacter. Our results indicate that 16S rRNA-based phylogeny does not provide sufficient species-level discrimination, since most of the unclassified Aminobacter strains belong to valid Aminobacter species or are putative new species. Since some members of the genus Aminobacter can utilize certain C1 compounds, such as methylamines and methyl halides, a comparative genomic analysis was performed to characterize the genetic basis of some degradative/assimilative pathways in the whole genus. Our findings suggest that all Aminobacter species are heterotrophic methylotrophs able to generate the methylene tetrahydrofolate intermediate through multiple oxidative pathways of C1 compounds and convey it in the serine cycle. Moreover, all Aminobacter species carry genes implicated in the degradation of phosphonates via the C-P lyase pathway, whereas only A. anthyllidis LMG 26462T contains a symbiosis island implicated in nodulation and nitrogen fixation.

The study investigated the effect of finisher diets containing two-stage fermented banana peel (FBP) meal on growth performance, haematological responses, intestinal microbiology and carcass traits of broilers. From days 22 to 38, 200 broiler chicks were grouped to maize-soybean meal-based finisher diet as a control (CONT) and finisher diets containing either 5% FBP (FBP-5), 10% FBP (FBP-10) or 15% FBP (FBP-15). Birds were blood sampled and slaughtered at day 38. For microbial population and villi height measurement, digesta and intestinal segments were collected. Breast meat was obtained for meat colour determination. Data collected were analysed according to analysis of variance followed by Duncan's multiple-range test when there was significant effect. There was no effect of treatments on broiler growth performance. The relative duodenum weight tended (P = 0.08) to be lower in FBP than in CONT. The mean corpuscular haemoglobin concentration (MCHC) was higher (P 0.05) in FBP-5 than in CONT and FBP-15. Uric acid concentration was lower (P 0.05) in FBP-10 and FBP-15 than in CONT. Feeding FBP tended (P = 0.08) to decrease coliform population in the ileum. Compared to CONT, the ileal villi heights were higher (P 0.05) and lower (P 0.05) in FBP-10 and FBP-5, respectively. The relative weight of thigh (P 0.05) and wings (P = 0.07) were higher in CONT than in FBP-fed birds, but the eviscerated carcass was not different (P 0.05) among groups. The lightness (L*) values of meat were higher (P 0.05) in FBP-10 and FBP-15 than in CONT and FBP-5. Compared with others, breast meat from FBP-10 birds had lower (P 0.05) redness (a*) values. In conclusion, dietary inclusion of FBP at the levels of up to 15% had no detrimental effect on growth and health performances of broiler chickens.

Objective: This study was aimed at evaluating the effect of fermentation using Chrysonillia crassa and Monascus purpureus on nutritional qualities, antioxidant, and antimicrobial activities of the used rice as a poultry feed ingredient. Materials and Methods: The used rice was soaked, steamed, and spread on a tray to cool. Suspension of M. purpureus or C. crassa was inoculated on the steamed used rice, and then mixed thoroughly. Afterward, the mixture was spread out on the tray, which was then covered with an aluminum foil. It was aerobically incubated for 7 and 4 days for the M. purpureus- and C. crassa-inoculated used rice, respectively. Subsequent to sun drying, the fermented used rice was grounded and analyzed. Results: Crude protein and ash contents were higher (p < 0.05) in the used rice fermented with C. crassa or M. purpureus than in the unfermented. Conversely, carbohydrate content was lower (p < 0.05) in the fermented compared with the unfermented. Gross energy and energy from fat were higher (p < 0.05) in the used rice fermented with M. purpureus than the unfermented. Amino acids L-methionine, L-serine, L-glutamic acid, L-valine glycine, L-leucine, L proline, L-threonine, L-histidine, and L-Sistine were higher (p < 0.05) in M. purpureus-fermented used rice than in C. crassa-fermented and the unfermented used rice. However, amino acids L-isoleucine, L- alanine, L-lysine, and L-tryptophan were higher (p < 0.05) in the used rice fermented with both C. crassa and M. purpureus, compared with the unfermented. L-tyrosine content was higher (p < 0.05) in M. purpureus-fermented used rice than in the unfermented. Furthermore, the antioxidant activities of the fermented products were higher (p < 0.05) than that of the unfermented. In addition, the antimicrobial activities of the fermented products against Staphylococcus aureus were higher (p < 0.05) than that of the unfermented used rice. Conclusion: In conclusion, the used rice fermented using C. crassa and M. purpureus improved the nutritional quality, as well as the antioxidant and antimicrobial activities of the products. [J Adv Vet Anim Res 2019; 6(2.000): 168-173]

Jeongeupia naejangsanensis is a Gram-negative, cellulose-degrading betaproteobacterium. Here, we report the draft genome sequence of the type strain J. naejangsanensis DSM 24253 and identify the genes implicated in the biosynthesis of polyhydroxyalkanoate bioplastic polymers.

Mutations in LRRK2 play a critical role in both familial and sporadic Parkinson&rsquo, s disease (PD). Up to date, the role of LRRK2 in PD onset and progression remains largely unknown. However, experimental evidence highlights a critical role of LRRK2 in the control of vesicle trafficking, likely by Rab phosphorylation, that in turn may regulate different aspects of neuronal physiology. Here we show that LRRK2 interacts with Sec8, one of eight subunits of the exocyst complex. The exocyst complex is an evolutionarily conserved multisubunit protein complex mainly involved in tethering secretory vesicles to the plasma membrane and implicated in the regulation of multiple biological processes modulated by vesicle trafficking. Interestingly, Rabs and exocyst complex belong to the same protein network. Our experimental evidence indicates that LRRK2 kinase activity or the presence of the LRRK2 kinase domain regulate the assembly of exocyst subunits and that the over-expression of Sec8 significantly rescues the LRRK2 G2019S mutant pathological effect. Our findings strongly suggest an interesting molecular mechanism by which LRRK2 could modulate vesicle trafficking and may have important implications to decode the complex role that LRRK2 plays in neuronal physiology.

// Silvio Daster 1, * , Nunzia Amatruda 1, 2, * , Diego Calabrese 2 , Robert Ivanek 2 , Eleonora Turrini 3 , Raoul A. Droeser 1 , Paul Zajac 1, 2 , Carmela Fimognari 3 , Giulio C. Spagnoli 1, 2 , Giandomenica Iezzi 1, 2 , Valentina Mele 1, 2, ** , Manuele G. Muraro 1, 2, ** 1 Department of Surgery, University Hospital Basel, Basel, Switzerland 2 Department of Biomedicine, University of Basel, Basel, Switzerland 3 Department for Life Quality Studies, University of Bologna, Rimini, Italy * These authors have contributed equally to this work ** Shared senior authorship Correspondence to: Valentina Mele, email: valentina.mele@usb.ch Manuele G. Muraro, email: manuele.muraro@usb.ch Keywords: multicellular tumor spheroids, three-dimensional culture, tumor model, hypoxia, necrosis Received: April 19, 2016 Accepted: September 19, 2016 Published: December 10, 2016 ABSTRACT Culture of cancerous cells in standard monolayer conditions poorly mirrors growth in three-dimensional architectures typically observed in a wide majority of cancers of different histological origin. Multicellular tumor spheroid (MCTS) culture models were developed to mimic these features. However, in vivo tumor growth is also characterized by the presence of ischemic and necrotic areas generated by oxygenation gradients and differential access to nutrients. Hypoxia and necrosis play key roles in tumor progression and resistance to treatment. To provide in vitro models recapitulating these events in highly controlled and standardized conditions, we have generated colorectal cancer (CRC) cell spheroids of different sizes and analyzed their gene expression profiles and sensitivity to treatment with 5FU, currently used in therapeutic protocols. Here we identify three MCTS stages, corresponding to defined spheroid sizes, characterized by normoxia, hypoxia, and hypoxia plus necrosis, respectively. Importantly, we show that MCTS including both hypoxic and necrotic areas most closely mimic gene expression profiles of in vivo-developing tumors and display the highest resistance to 5FU. Taken together, our data indicate that MCTS may mimic in vitro generation of ischemic and necrotic areas in highly standardized and controlled conditions, thereby qualifying as relevant models for drug screening purposes.

Increasing evidence supports the anti-inflammatory role of estrogens in Multiple Sclerosis (MS), originating from the observation of reduction in relapse rates among women with MS during pregnancy, but the molecular mechanisms are still not completely understood. Using an integrative data analysis, we identified T helper (Th) 17 and T regulatory (Treg) cell-type-specific regulatory regions (CSR) regulated by estrogen receptor alpha (ERα). These CSRs were validated in polarized Th17 from healthy donors (HD) and in peripheral blood mononuclear cells, Th17 and Treg cells from relapsing remitting (RR) MS patients and HD during pregnancy. 17β-estradiol induces active histone marks enrichment at Forkhead Box P3 (FOXP3)-CSRs and repressive histone marks enrichment at RAR related orphan receptor C (RORC)-CSRs in polarized Th17 cells. A disease-associated epigenetic profile was found in RRMS patients during pregnancy, suggesting a FOXP3 positive regulation and a RORC negative regulation in the third trimester of pregnancy. Altogether, these data indicate that estrogens act as immunomodulatory factors on the epigenomes of CD4+ T cells in RRMS; the identified CSRs may represent potential biomarkers for monitoring disease progression or new potential therapeutic targets.

Anemia of β-thalassemia is caused by ineffective erythropoiesis and reduced red cell survival. Several lines of evidence indicate that iron/heme restriction is a potential therapeutic strategy for the disease. Glycine is a key initial substrate for heme and globin synthesis. We provide evidence that bitopertin, a glycine transport inhibitor administered orally, improves anemia, reduces hemolysis, diminishes ineffective erythropoiesis, and increases red cell survival in a mouse model of β-thalassemia (Hbb(th3/+) mice). Bitopertin ameliorates erythroid oxidant damage, as indicated by a reduction in membrane-associated free α-globin chain aggregates, in reactive oxygen species cellular content, in membrane-bound hemichromes, and in heme-regulated inhibitor activation and eIF2α phosphorylation. The improvement of β-thalassemic ineffective erythropoiesis is associated with diminished mTOR activation and Rab5, Lamp1, and p62 accumulation, indicating an improved autophagy. Bitopertin also upregulates liver hepcidin and diminishes liver iron overload. The hematologic improvements achieved by bitopertin are blunted by the concomitant administration of the iron chelator deferiprone, suggesting that an excessive restriction of iron availability might negate the beneficial effects of bitopertin. These data provide important and clinically relevant insights into glycine restriction and reduced heme synthesis strategies for the treatment of β-thalassemia.

The use of herbal food supplements, as a concentrate form of vegetable extracts, increased so much over the past years to count them among the relevant sources of dietetic polyphenols. Bud-derivatives are a category of botanicals perceived as a &ldquo, new entry&rdquo, in this sector since they are still poorly studied. Due to the lack of a manufacturing process specification, very different products can be found on the market in terms of their polyphenolic profile depending on the experimental conditions of manufacturing. In this research two different manufacturing processes, using two different protocols, and eight species (Carpinus betulus L., Cornus mas L., Ficus carica L., Fraxinus excelsior L., Larix decidua Mill., Pinus montana Mill., Quercus petraea (Matt.) Liebl., Tilia tomentosa Moench), commonly used to produce bud-derivatives, have been considered as a case study. An untargeted spectroscopic fingerprint of the extracts, coupled to chemometrics, provide to be a useful tool to identify these botanicals. The targeted phytochemical fingerprint by HPLC provided a screening of the main bud-derivatives polyphenolic classes highlighting a high variability depending on both method and protocol used. Nevertheless, ultrasonic extraction proved to be less sensitive to the different extraction protocols than conventional maceration regarding the extract polyphenolic profile.

Resistance to antimalarial drugs has spread rapidly over the past few decades. The WHO recommends artemisinin-based combination therapies for the treatment of uncomplicated malaria, but unfortunately these approaches are losing their efficacy in large areas of Southeast Asia. In 2016, artemisinin resistance was confirmed in 5 countries of the Greater Mekong subregion. We focused our study on Syk inhibitors as antimalarial drugs. The Syk protein is present in human erythrocytes, and the membrane of protein band 3 is its major target following activation by oxidant stress. Tyr phosphorylation of band 3 occurs during P. falciparum growth, leading to the release of microparticles containing hemicromes and structural weakening of the host cell membrane, simplifying merozoite reinfection. Syk inhibitors block these events by interacting with the Syk protein&rsquo, s catalytic site. We performed in vitro proteomics and in silico studies and compared the results. In vitro studies were based on treatment of the parasite&rsquo, s cellular cultures with different concentrations of Syk inhibitors, while proteomics studies were focused on the Tyr phosphorylation of band 3 by Syk protein with the same concentrations of drugs. In silico studies were based on different molecular modeling approaches in order to analyze and optimize the ligand&ndash, protein interactions and obtain the highest efficacy in vitro. In the presence of Syk inhibitors, we observed a marked decrease of band 3 Tyr phosphorylation according to the increase of the drug&rsquo, s concentration. Our studies could be useful for the structural optimization of these compounds and for the design of novel Syk inhibitors in the future.

This study aimed to compare the antimicrobial action of three soaps for hand hygiene (HH): 2.0% Tea Tree Oil (TTO); 0.5% triclosan; 2.0% chlorhexidine, and to explore the perception of healthcare professionals about TTO. Two-step study: a quantitative, to determine the logarithmic reduction of Escherichia coli K12 colony-forming units before and after HH of 15 volunteers and quali-quantitative, through interviews with 23 health professionals. All the three products demonstrated antimicrobial action (a log10 reduction factor of 4.18 for TTO, 4.31 for triclosan, 3.89 for chlorhexidine, and 3.17 for reference soap). Professionals remarked the pleasant aroma and non-dryness of skin when using soap containing TTO.

International audience; Although artemisinin-based combination therapies (ACTs) treat Plasmodium falciparum malaria effectively throughout most of the world, the recent expansion of ACT-resistant strains in some countries of the Greater Mekong Subregion (GMS) further increased the interest in improving the effectiveness of treatment and counteracting resistance. Recognizing that (1) partially denatured hemoglobin containing reactive iron (hemichromes) is generated in parasitized red blood cells (pRBC) by oxidative stress, (2) redox-active hemichromes have the potential to enhance oxidative stress triggered by the parasite and the activation of artemisinin to its pharmaceutically active form, and (3) Syk kinase inhibitors block the release of membrane microparticles containing hemichromes, we hypothesized that increasing hemichrome content in parasitized erythrocytes through the inhibition of Syk kinase might trigger a virtuous cycle involving the activation of artemisinin, the enhancement of oxidative stress elicited by activated artemisinin, and a further increase in hemichrome production. We demonstrate here that artemisinin indeed augments oxidative stress within parasitized RBCs and that Syk kinase inhibitors further increase iron-dependent oxidative stress, synergizing with artemisinin in killing the parasite. We then demonstrate that Syk kinase inhibitors achieve this oxidative enhancement by preventing parasite-induced release of erythrocyte-derived microparticles containing redox-active hemichromes. We also observe that Syk kinase inhibitors do not promote oxidative toxicity to healthy RBCs as they do not produce appreciable amounts of hemichromes. Since some Syk kinase inhibitors can be taken daily with minimal side effects, we propose that Syk kinase inhibitors could evidently contribute to the potentiation of ACTs.

Mycobacterium vaccae is a rapidly growing nonpathogenic species of the Mycobacteriaceae family of bacteria that can cause pulmonary and disseminated disease in particular in immunocompromised individuals. Here we describe a first case of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass-spectrometry (MS) identification of this pathogen in a patient with non-Hodgkin’s lymphoma during chemoimmunotherapy salvage treatment, and its impact on clinical decision making.

Many hypotheses have been proposed to explain how a glutamate to valine substitution in sickle haemoglobin (HbS) can cause sickle cell disease (SCD). We propose and document a new mechanism in which elevated tyrosine phosphorylation of Band 3 initiates sequelae that cause vaso-occlusion and the symptoms of SCD. In this mechanism, denaturation of HbS and release of heme generate intracellular oxidants which cause inhibition of erythrocyte tyrosine phosphatases, thus permitting constitutive tyrosine phosphorylation of Band 3. This phosphorylation in turn induces dissociation of the spectrin-actin cytoskeleton from the membrane, leading to membrane weakening, discharge of membrane-derived microparticles (which initiate the coagulation cascade) and release of cell-free HbS (which consumes nitric oxide) and activates the endothelium to express adhesion receptors). These processes promote vaso-occlusive events which cause SCD. We further show that inhibitors of Syk tyrosine kinase block Band 3 tyrosine phosphorylation, prevent release of cell-free Hb, inhibit discharge of membrane-derived microparticles, increase sickle cell deformability, reduce sickle cell adhesion to human endothelial cells, and enhance sickle cell flow through microcapillaries. In view of reports that imatinib (a Syk inhibitor) successfully treats symptoms of sickle cell disease, we suggest that Syk tyrosine kinase inhibitors warrant repurposing as potential treatments for SCD.

Vermicomposting is a natural process that utilizes earthworms and associated microbiome to transform organic wastes into vermicompost by-products that are rich in beneficial microorganisms and nutrients such as carbon, phosphorus, nitrogen, magnesium, and calcium. Liquid vermicompost extract (LVE), a derivative of the vermicomposting process, has recently gained interest among scientists and organic farmers due to their potential ability to enhance tripartite plant-microbe-soil interactions that would lead to improved plant and root growth, soil health and overall crop yield productivity. To investigate the short-term effect of LVE on soil mycorrhizal inoculum potential (MIP) and plant-mycobiome interactions, a field trial was carried out at CiRAA E. Avanzi, San Piero a Grado, Pisa, Italy.The effect of LVE and its associated microbial and chemical components on soil MIP and AMF root colonization was evaluated on five summer crops, i.e. chickpea (Cicer arietinum L.), berseem clover (Trifolium alexandrinum L.), lentil (Lens culinaris L.), soybean (Glycine max L. Merrill), and sunflower (Helianthus annuus L.). The test plants were grown with or without the application of LVE in a split-plot trial with five replicates. Freshly made LVE from vermicomposting of wheat straws mixed with horse manure was screened for microbial properties using the Illumina Miseq sequencing platform. Seed inoculation with LVE was done before planting while field inoculation was done at the stem-elongation stage. Un-inoculated seeds and plots were used as controls. Soil MIP was assessed before planting and after harvesting, while AMF root colonization was evaluated at the mid-flowering stage of each crop.The bacterial 16S and fungal ITS sequence analyses showed a high bacteria and fungal abundance and taxonomic alpha diversity present in the LVE. The most dominant taxa included Mucor, Citrobacter, Pseudomonas, Arcobacter, Azomonas and Clostridium. These microbes are commonly found in agricultural soil and are linked to the hydrolysis of complex organic matter, nutrient recycling, production of growth-promoting factors and siderophores, while others are known to produce peptide antimycotics and antibiotics that protect plants against pathogenic soil microorganisms.The soil MIP significantly (p < 0.0001) differed between the two soil sampling times (before planting and after harvesting). It was evident that both seed and field inoculation with LVE significantly enhanced the soil MIP and this could benefit the next crop under rotation. AMF root colonization varied significantly across the crop species (p < 0.0001) and LVE treatment (p = 0.006). Highly nodulated lentils and berseem clover roots recorded significantly higher AMF root colonization than all the other crops. LVE inoculation had an overall positive effect on AMF root colonization with an average increase of 6.2% compared to the un-inoculated crops. These short-term results indicate that there could be a positive effect of the LVE inoculation on the soil MIP and AMF root colonization of our test crops, which could be attributed to the beneficial additive effects of the LVE that enhanced the tripartite plant-microbe-soil interactions. Keywords: Vermicomposting, Liquid vermicompost extract, Soil MIP, AMF root colonization, Legumes

Arbuscular mycorrhizal fungi (AMF) establish mutualistic symbioses with the roots of most plant species, playing a key role in crop productivity and ecosystem functioning. The characterization of local AMF genotypes assemblages is a prerequisite for the reproduction, both ex situ and on farm, of efficient native AMF communities, in order to improve soil quality and fertility through agricultural management and to use selected AMF as inoculants in sustainable agriculture. Contrasting evidences have been found by studies addressing the influence of different environmental drivers, among which geographical distance, on the diversity and composition of AMF communities in agricultural soils. In the present study, a Mediterranean “hot-spot” of AMF species richness was studied to assess whether AMF display a spatial structure at a local scale and whether their spore distribution is affected by soil variables. AMF distribution patterns were estimated by spore analysis, in 12 field plots, which were characterized for their chemical and physical variables. Plots were distributed along a transect across the field, and sampling points distances ranged from 4 to 102 m. While most taxa displayed an aggregated distribution, no significant spatial correlation was detected among AMF communities. On the contrary, AMF communities were significantly correlated with the entire set of soil variables, and with distinct soil characteristics, such as pH and total N, but not P. Among the most abundant taxa, Funneliformis geosporus, Septoglomus constrictum, Funneliformis mosseae, Ambispora granatensis and Glomus badium, were positively affected by total N, and Paraglomus occultum negatively affected by pH. Multiple regression analysis revealed that distribution of most AMF species was significantly related to nitrogen, clay, cation exchange capacity and pH. Multivariate (RDA) analysis confirmed results of multiple regression, with the best model selecting three soil variables (N, clay and pH). Variance partitioning analysis also showed that AMF distribution was mostly explained by environmental variables (36.4%) and by environmental variables spatial structure (16.4%). Overall, the results of this work revealed the importance of soil variables in shaping AMF community composition and spatial heterogeneity, within a site characterized by high AMF richness.

Each one of us in his life has, at least once, smelled the scent of roses, read one canto of Dante’s Commedia or listened to the sound of the sea from a shell. All of this is possible thanks to the astonishing capabilities of an organ, such as the brain, that allows us to collect and organize perceptions coming from sensory organs and to produce behavioural responses accordingly. Studying an operating brain in a non-invasive way is extremely difficult in mammals, and particularly in humans. In the last decade, a small teleost fish, zebrafish (Danio rerio), has been making its way into the field of neurosciences. The brain of a larval zebrafish is made up of 'only' 100000 neurons and it’s completely transparent, making it possible to optically access it. Here, taking advantage of the best of currently available technology, we devised optical solutions to investigate the dynamics of neuronal activity throughout the entire brain of zebrafish larvae.

Arbuscular mycorrhizal fungi (AMF) are a key group of beneficial obligate biotrophs, establishing a mutualistic symbiosis with the roots of most land plants. The molecular markers generally used for their characterization are mainly based on informative regions of nuclear rDNA (SSU-ITS-LSU), although protein-encoding genes have also been proposed. Within functional genes, those encoding for phosphate transporters (PT) are particularly important in AMF, given their primary ability to take up Pi from soil, and to differentially affect plant phosphate nutrition. In this work, we investigated the genetic diversity of PT1 gene sequences and sequences of the taxonomically relevant SSU-ITS-LSU region in two isolates of the species Funneliformis coronatus, three isolates of the species Funneliformis mosseae and two species of the genus Rhizoglomus, originated from geographically distant areas and cultured in vivo. Our results showed that partial PT1 sequences not only successfully differentiated AMF genera and species like ribosomal gene sequences but also highlighted intraspecific diversity among F. mosseae and F. coronatus isolates. The study of functional genes related to the uptake of key mineral nutrients for the assessment of AMF diversity represents a key step in the selection of efficient isolates to be used as inocula in sustainable agriculture.

Background: Sickle cell disease (SCD) derives from a mutation in sickle hemoglobin (HbS) that leads to vaso-occlusive events that eventually cause organ failure. Based on observations from multiple labs, we hypothesize that the events linking HbS denaturation to vaso-occlusion are: HbS denaturation→oxidative stress→inhibition of erythrocyte tyrosine phosphatases→enhanced tyrosine phosphorylation of band 3→dissociation of the spectrin/actin cytoskeleton from the membrane→release of membrane-derived microparticles (MPs) and discharge of cell-free HbS→induction of microthromboses and activation of the vascular endothelium→vascular occlusion. In the studies below, we document each step of this pathway and demonstrate that inhibitors of Band 3 tyrosine phosphorylation block all downstream events. Methods: Band 3 tyrosine phosphorylation, plasma MP counts, plasma cell-free Hb concentration, sickle cell deformability, sickle cell flow through micro-capillaries, and adhesion of sickle erythrocytes to activated endothelial cells are quantitated using fresh sickle blood both before and after incubation with inhibitors of Band 3 tyrosine phosphorylation. Findings: Imatinib and other Syk kinase inhibitors block Band 3 tyrosine phosphorylation, suppress release of membrane MPs, reduce discharge of cell-free HbS, improve sickle cell deformability, enhance flow of sickle cells through microcapillaries, and reduce adherence of sickle cells to heme-activated endothelium. Interpretation: Inhibition of Band 3 Tyrosine phosphorylation restores membrane integrity and thereby suppresses the downstream sequelae that lead to vaso-occlusive events in SCD. Inhibitors of Band 3 tyrosine phosphorylation may therefore constitute a novel effective therapy for SCD. Funding Statement: NIH grants R01GM24417-40 (P.S.L.), 1RF1 NS110049-01 (J.W.), R01HL133574 (U.A.G. and J.A.L.), T32HL134622 (R.A.), and NSF #1552782 (U.A.G.). Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: All sickle cell blood samples were obtained following informed consent using procedures approved by the local IRBs.

Although current malaria therapies inhibit pathways encoded in the parasite’s genome, we have looked for anti-malaria drugs that can target an erythrocyte component because development of drug resistance might be suppressed if the parasite cannot mutate the drug’s target. In search for such erythrocyte targets, we noted that human erythrocytes express tyrosine kinases, whereas thePlasmodium falciparumgenome encodes no obvious tyrosine kinases. We therefore screened a library of tyrosine kinase inhibitors from Eli Lilly and Co. in a search for inhibitors with possible antimalarial activity. We report that although most tyrosine kinase inhibitors exerted no effect on parasite survival, a subset of tyrosine kinase inhibitors displayed potent anti-malarial activity. Moreover, all inhibitors found to block tyrosine phosphorylation of band 3 specifically suppressedP.falciparumsurvival at the parasite egress stage of its intra-erythrocyte life cycle. Conversely, tyrosine kinase inhibitors that failed to block band 3 tyrosine phosphorylation but still terminated the parasitemia were observed to halt parasite proliferation at other stages of the parasite’s life cycle. Taken together these results suggest that certain erythrocyte tyrosine kinases may be important toP.falciparummaturation and that inhibitors that block these kinases may contribute to novel therapies forP.falciparummalaria.