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Abstract Background Emergency department telepsychiatry (EDTP) can improve healthcare delivery. However, no studies document implementation and overall satisfaction within the Canadian pandemic context. The objectives of this study were to (i) describe barriers associated with EDTP; and (ii) assess patients’, ED health professionals’, and psychiatrists’ perceived satisfaction with EDTP during the COVID-19 pandemic. Data were collected in three New Brunswick hospitals over 8-weeks in 2021. Psychiatrists (n = 6) completed a self-report questionnaire following each EDTP consultation, patients (n = 58) completed a telephone-administered questionnaire 1-week post EDTP consultation, and ED health professionals (n = 31) completed a single self-report questionnaire at the end of the study period. Results One-third of psychiatrists and two-thirds of ED health professionals encountered a barrier, respectively. The most common barriers related to problems linked to sound/video/connection, lack of experience and guidelines, or increased workload. Despite these barriers, high levels of satisfaction were reported by patients and psychiatrists (mean satisfaction score (sd): 4.2 (0.6) and 4.8 (0.4), respectively). ED health professionals reported lower satisfaction scores; however, most believed that EDTP was associated with healthcare delivery advantages. Conclusions These findings indicate moderate to high EDTP satisfaction. Additional training, guidelines, and change management strategies may be necessary to insure harmonious EDTP implementation for all health professionals.

This study investigated reasons for lower-than-expected uptake of germline genetic testing compared to national guidelines amongst adult patients with cancer, self-reporting clinician recommendation for genetic testing. Cross-sectional survey of 596 patients with a personal history of cancer, responded about their cancer diagnosis, physician recommendation for and status of genetic testing and demographics. Adjusting for potential confounding factors (cancer type, education, income and insurance status) male sex significantly decreased odds of receiving a clinical recommendation for genetic testing (Odds Ratio: 0.06; 95% Confidence Interval: 0.04–0.10). Females, with a diagnosis of breast cancer, were more likely to receive a recommendation than other cancer types (64.8% vs. 90.9%, p < 0.001). Participants who received a physician recommendation were significantly more likely to receive genetic testing (p < 0.001). Clinician recommendation is an important driver of genetic testing, necessitating efforts to increase clinician recommendations, particularly for males and patients with cancers other than breast.

Abstract Background Family-centered rounds is recognized as a best practice for hospitalized children, but it has only been possible for children whose families can physically be at the bedside during hospital rounds. The use of telehealth to bring a family member virtually to the child’s bedside during hospital rounds is a promising solution. We aim to evaluate the impact of virtual family-centered hospital rounds in the neonatal intensive care unit on parental and neonatal outcomes. Methods This two-arm cluster randomized controlled trial will randomize families of hospitalized infants to have the option to use telehealth for virtual hospital rounds (intervention) or usual care (control). The intervention-arm families will also have the option to participate in hospital rounds in-person or to not participate in hospital rounds. All eligible infants who are admitted to this single-site neonatal intensive care unit during the study period will be included. Eligibility requires that there be an English-proficient adult parent or guardian. We will measure participant-level outcome data to test the impact on family-centered rounds attendance, parent experience, family-centered care, parent activation, parent health-related quality of life, length of stay, breastmilk feeding, and neonatal growth. Additionally, we will conduct a mixed methods implementation evaluation using the RE-AIM (Reach, Effectiveness, Adoption, Implementation, Maintenance) framework. Discussion The findings from this trial will increase our understanding about virtual family-centered hospital rounds in the neonatal intensive care unit. The mixed methods implementation evaluation will enhance our understanding about the contextual factors that influence the implementation and rigorous evaluation of our intervention. Trial registration ClinicalTrials.gov Identifier: NCT05762835. Status: Not yet recruiting. First posted: March 10, 2023; last update posted: March 10, 2023.

Abstract Background Only a small proportion of patients who qualify for clinical genetic testing for cancer susceptibility get testing. Many patient‐level barriers contribute to low uptake. In this study, we examined self‐reported patient barriers and motivators for cancer genetic testing. Methods A survey comprised of both new and existing measures related to barriers and motivators to genetic testing was emailed to patients with a diagnosis of cancer at a large academic medical center. Patients who self‐reported receiving a genetic test were included in these analyses (n = 376). Responses about emotions following testing as well as barriers and motivators prior to getting testing were examined. Group differences in barriers and motivators by patient demographic characteristics were examined. Results Being assigned female at birth was associated with increased emotional, insurance, and family concerns as well as increased health benefits compared to patients assigned male at birth. Younger respondents had significantly higher emotional and family concerns compared to older respondents. Recently diagnosed respondents expressed fewer concerns about insurance implications and emotional concerns. Those with a BRCA‐related cancer had higher scores on social and interpersonal concerns scale than those with other cancers. Participants with higher depression scores indicated increased emotional, social and interpersonal, and family concerns. Conclusions Self‐reported depression emerged as the most consistent factor influencing report of barriers to genetic testing. By incorporating mental health resources into clinical practice, oncologists may better identify those patients who might need more assistance following through with a referral for genetic testing and the response afterwards.

Seed germination is critical for seedling procedures, and new technologies are required to increase crop yields. Some substances are considered elicitors or stimulating agents because they promote seed germination as well as plant growth (like root and plumule length, and germination rate among others). Some of these, could be applied during plant growth, or in earlier phases before germination as seeds pretreatment. Neutral electrolyzed water (NEW) is a solution of oxidizing agents mainly studied for its antimicrobial power mainly applied to food pathogens, and in less reports to phytopathogens. The objective of this work is to evaluate the effect of NEW on germination and morphological parameters of tomato (Lycopersicon esculentum), cucumber (Cucumis sativus) and lettuce (Lactuca sativa L.) seeds as well as to evaluate the in vitro activity of NEW against F. oxysporum. Two experiments were carried out to determine the in vitro and in vivo effects of NEW in seed pretreatment and another in vitro experiment was also carried out to determine antifungal effect against F. oxysporum. NEW in vitro treatments at 50 mg/L decreased spore concentration greater than 4 log spores/mL. Germination parameters and morphological variables were analyzed after NEW pretreatments. For in vitro germination, NEW pretreatments up to 75 mg/L did not show effect on germination parameters however highest dose was slightly adverse (4 % against water control). For in vivo germination, NEW pretreatment of 100 mg/L had a stimulating effect on the germination rate in cucumber seeds (100 % FGP against 83 % FGP for water control). For tomato seeds germination neither stimulate nor adverse effect was observed (same percentage as water control). For lettuce seeds germination an inhibitory effect for 12 and 24 h of exposure time was achieved (73 % NEW treatment against 90 % of water control). For tomato and lettuce seeds, the NEW pretreatment of 50 mg/L during 12 h increased its germination capacity.

IntroductionThe COVID-19 pandemic presented health systems across the globe with unparalleled socio-political, ethical, scientific, and economic challenges. Despite the necessity for a unified, innovative, and effective response, many jurisdictions were unprepared to such a profound health crisis. This study aims to outline the creation of an evaluative tool designed to measure and evaluate the Vitalité Health Network’s (New Brunswick, Canada) ability to manage health crises.MethodsThe methodology of this work was carried out in four stages: (1) construction of an evaluative framework; (2) validation of the framework; (3) construction of the evaluative tool for the Health Authority; and (4) evaluation of the capacity to manage a health crisis.ResultsThe resulting evaluative tool incorporated 8 dimensions, 74 strategies, and 109 observable elements. The dimensions included: (1) clinical care management; (2) infection prevention and control; (3) governance and leadership; (4) human and logistic resources; (5) communication and technologies; (6) health research; (7) ethics and values; and (8) training. A Canadian Health Authority implemented the tool to support its future preparedness.ConclusionThis study introduces a methodological strategy adopted by a Canadian health authority to evaluate its capacity in managing health crises. Notably, this study marks the first instance where a Canadian health authority has created a tool for emergency healthcare management, informed by literature in the field and their direct experience from handling the SARS-CoV-2 pandemic.

Abstract Background Although most cancers are sporadic, germline genetic variants are implicated in 5–10% of cancer cases. Clinical genetic testing identifies pathogenic germline genetic variants for hereditary cancers. The Michigan Genetic Hereditary Testing (MiGHT) study is a three-arm randomized clinical trial that aims to test the efficacy of two patient-level behavioral interventions on uptake of cancer genetic testing. Methods The two interventions being tested are (1) a virtual genetics navigator and (2) motivational interviewing by genetic health coaches. Eligible participants are adults with a diagnosis of breast, prostate, endometrial, ovarian, colorectal, or pancreatic cancer who meet the National Comprehensive Cancer Network (NCCN) criteria for genetic testing. Participants are recruited through community oncology practices affiliated with the Michigan Oncology Quality Consortium (MOQC) and have used the Family Health History Tool (FHHT) to determine testing eligibility. The recruitment goal is 759 participants, who will be randomized to usual care or to either the virtual genetics navigator or the motivational interviewing intervention arms. The primary outcome will be the proportion of individuals who complete germline genetic testing within 6 months. Discussion This study addresses patient-level factors which are associated with the uptake of genetic testing. The study will test two different intervention approaches, both of which can help address the shortage of genetic counselors and improve access to care. Trial registration This study has been approved by the Institutional Review Board of the University of Michigan Medical School (HUM00192898) and registered in ClinicalTrials.gov (NCT05162846).

Abstract Several factors are associated with the severity of the respiratory disease caused by the influenza virus. Although viral factors are one of the most studied, in recent years the role of the microbiota and co-infections in severe and fatal outcomes has been recognized. However, most of the work has focused on the microbiota of the upper respiratory tract (URT), hindering potential insights from the lower respiratory tract (LRT) that may help to understand the role of the microbiota in Influenza disease. In this work, we characterized the microbiota of the LRT of patients with Influenza A using 16S rRNA sequencing. We tested if patients with different outcomes (deceased/recovered) and use of antibiotics differ in their microbial community composition. We found important differences in the diversity and composition of the microbiota between deceased and recovered patients. In particular, we detected a high abundance of opportunistic pathogens such as Granulicatella, in patients either deceased or with antibiotic treatment. Also, we found antibiotic treatment correlated with lower diversity of microbial communities and with lower probability of survival in Influenza A patients. Altogether, the loss of microbial diversity could generate a disequilibrium in the community, potentially compromising the immune response increasing viral infectivity, promoting the growth of potentially pathogenic bacteria that, together with altered biochemical parameters, can be leading to severe forms of the disease. Overall, the present study gives one of the first characterizations of the diversity and composition of microbial communities in the LRT of Influenza patients and its relationship with clinical variables and disease severity.

IntroductionWe examined whether factors identified as associated with cannabis use at age 14 to 16 years are also associated with ever use at age 12. MethodsParticipants in the AdoQuest study (n = 1852) were recruited in 2005 from among Grade 5 students in 29 French-language elementary schools in Montréal, Canada. Self-report data were collected from participants in Grade 5 (spring 2005) and 6 (fall 2005 and spring 2006) and from parents/guardians in 2006/07. Inclusion in the analytic sample (n = 1076; mean age [SD] = 10.7 [0.5]) required data from participant and parental questionnaires and data on cannabis use in Grade 6 (mean age [SD] = 11.7 [0.4]). We estimated associations between ever use at age 12 with 33 potential correlates, separately in unadjusted and adjusted logistic regression models. ResultsFifty-three participants (4.9%) reported ever use at age 12. Factors associated with higher odds of ever use included older age, identifying as male, lower household income, more weekly spending money, ever tried cigarettes or other tobacco products, ever drank alcohol or binge drank, ever gambled, friends or siblings smoke cigarettes, greater nicotine dependence, higher depressive symptoms and greater impulsivity. Protective factors included higher levels of parental/guardian monitoring and greater self-esteem and school connectedness. ConclusionFactors associated with cannabis use at later ages are also associated with ever use at age 12. Our findings suggest that surveillance for and interventions to prevent cannabis use are warranted before age 12.

IntroductionNous avons vérifié si les facteurs connus pour être associés à la consommation de cannabis entre 14 et 16 ans sont aussi associés au fait d’avoir déjà consommé du cannabis à l’âge de 12 ans. MéthodologieLes participants à l’étude AdoQuest (n = 1 852) ont été recrutés en 2005 parmi les élèves de 5e année de 29 écoles primaires francophones à Montréal (Canada). Des données autodéclarées ont été recueillies auprès des participants en 5e année (printemps 2005) et en 6e année (automne 2005 et printemps 2006) ainsi qu’auprès de leurs parents ou personnes responsables en 2006-2007. Pour que les participants soient inclus dans l’échantillon d’analyse (n = 1 076; âge médian = 10,7 ans [écart-type : 0,5]), il fallait avoir accès aux données des questionnaires remplis par les participants et par leurs parents de même qu’aux données sur leur consommation de cannabis en 6e année (âge médian = 11,7 ans [écart-type : 0,4]). Nous avons estimé les associations possibles entre le fait d’avoir déjà consommé du cannabis à 12 ans et 33 facteurs de corrélation évalués séparément dans des modèles de régression logistique ajustés et non ajustés. RésultatsCinquante-trois participants (4,9 %) ont déclaré avoir déjà consommé du cannabis à l’âge de 12 ans. Les facteurs associés à une consommation étaient un âge plus élevé, s’identifier comme garçon, un faible revenu du ménage, une allocation hebdomadaire élevée, le fait d’avoir déjà consommé des cigarettes ou d’autres produits du tabac, le fait d’avoir déjà bu de l’alcool, le fait d’avoir déjà eu une consommation excessive d’alcool, le fait d’avoir déjà joué à des jeux de hasard, le tabagisme chez les amis ou les frères et soeurs, une dépendance accrue à la nicotine, des symptômes de dépression importants et une forte impulsivité. Les facteurs de protection étaient un niveau élevé de surveillance par les parents ou les personnes responsables des jeunes, une bonne estime de soi et un sentiment d’appartenance à l’école. ConclusionLes facteurs associés à la consommation de cannabis à des âges plus élevés sont aussi associés au fait d’avoir déjà consommé du cannabis à l’âge de 12 ans. D’après nos résultats, une surveillance et des interventions visant à prévenir la consommation de cannabis sont nécessaires chez les jeunes de moins de 12 ans.

In recent years, the demand of healthier food products and products made with natural ingredients has increased overwhelmingly, led by the awareness of human beings of the influence of food on their health, as well as by the evidence of side effects generated by different ingredients such as some additives. This is the case for several artificial colorants, especially azo colorants, which have been related to the development of allergic reactions, attention deficit and hyperactivity disorder. All the above has focused the attention of researchers on obtaining colorants from natural sources that do not present a risk for consumption and, on the contrary, show biological activity. The most representative compounds that present colorant capacity found in nature are anthocyanins, anthraquinones, betalains, carotenoids and chlorophylls. Therefore, the present review summarizes research published in the last 15 years (2008–2023) in different databases (PubMed, Scopus, Web of Science and ScienceDirect) encompassing various natural sources of these colorant compounds, referring to their obtention, identification, some of the efforts made for improvements in their stability and their incorporation in different food matrices. In this way, this review evidences the promising path of development of natural colorants for the replacement of their artificial counterparts.

Shiga toxin-producing Escherichia coli (STEC) O157:H7 (O157) is a foodborne pathogen causing human disease ranging from hemorrhagic colitis and hemolytic uremic syndrome to kidney failure, while remaining harmless to cattle, its primary reservoir. The severity of the human disease associated mainly with Shiga toxin production and a global emergence of antibiotic resistant STEC highlights the need for effective non-antibiotic, pre-harvest strategies to reduce O157 in cattle, the principal source of human infection. Towards this goal three synthetic antimicrobial peptides (AMPs): human granulysin (hGRNL), bovine NK-lysin (bNK2A), and porcine NK-lysin (pNKL), were tested in vitro against O157 isolates. As expected, circular dichroism spectroscopy findings were consistent with a predominantly α-helical conformation for all three AMPs in an environment mimicking bacterial outer surface or liposaccharides. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations of hGRNL (200 μM), bNK2A (12.5 μM against strain 86-24 and 25 μM against EDL933), and pNKL (6.25 μM) were determined using the Clinical and Laboratory Standards Institute broth microdilution method in Müeller-Hinton broth (cation-adjusted). The bNK2A and pNKL AMPs did not induce Shiga toxin expression in O157 at MIC, as there was a significant decrease or no change in toxin expression following 4- or 20 h incubation with the AMPs; bNK2A p

Soft rot disease causes devastating losses to crop plants all over the world, with up to 90% loss in tropical climates. To better understand this economically important disease, we isolated four soft rot-causing Erwinia persicina strains from rotted vegetables. Notably, E. persicina has only recently been identified as a soft rot pathogen and a comprehensive genomic analysis and comparison has yet to be conducted. Here, we provide the first genomic analysis of E. persicina, compared to Pectobacterium carotovorum, P. carotovorum, and associated Erwinia plant pathogens. We found that E. persicina shares common genomic features with other Erwinia species and P. carotovorum, while having its own unique characteristics as well. The E. persicina strains examined here lack Type II and Type III secretion systems, commonly used to secrete pectolytic enzymes and evade the host immune response, respectively. E. persicina contains fewer putative pectolytic enzymes than P. carotovorum and lacks the Out cluster of the Type II secretion system while harboring a siderophore that causes a unique pink pigmentation during soft rot infections. Interestingly, a putative phenolic acid decarboxylase is present in the E. persicina strains and some soft rot pathogens, but absent in other Erwinia species, thus potentially providing an important factor for soft rot. All four E. persicina isolates obtained here and many other E. persicina genomes contain plasmids larger than 100 kbp that encode proteins likely important for adaptation to plant hosts. This research provides new insights into the possible mechanisms of soft rot disease by E. persicina and potential targets for diagnostic tools and control measures.

Objective Canadian fetal alcohol spectrum disorder (FASD) guidelines encourage an age-specific interdisciplinary diagnostic approach. However, there is currently no standard-of-care regarding FASD diagnosis disclosure and few studies document Canadian FASD clinical capacity. Our objectives were to describe clinical capacity (defined as skills and resources) for FASD assessment, diagnosis, disclosure and support in Canada.Design, setting and participants Data were drawn from the CanDiD study, a cross-sectional investigation of Canadian FASD clinical capacity. Forty-one clinics participated in the study. Data were collected in 2021 on the number and types of health professionals included in the assessment and diagnostic teams, the presence (or absence) of a minor patient when the FASD diagnosis is disclosed to parents/guardians, who is responsible for the diagnosis disclosure, the use of explanatory tools, and the types of support/counselling services available. The proportion of clinics that follow the Canadian interdisciplinary diagnostic guidelines by age group is described among participating clinics.Results Overall, 21, 13 and 7 specialised FASD clinics were in Western/Northern, Central and Atlantic Canada, respectively. The number of referrals per year surpassed the number of diagnostic assessments completed in all regions. Approximately, 60% of clinics who diagnosed FASD in infants and preschool children (n=4/7 and 15/25, respectively) followed the interdisciplinary guidelines compared with 80% (n=32/40) in clinics who diagnosed school-aged children/adolescents. Diagnostic reporting practices were heterogeneous, but most used an explanatory tool with children/adolescents (67%), offered support/counselling (90–95%) and used case-by-case approach (80%) when deciding who would disclose the diagnosis to the child/adolescent and when.Conclusions Limited diagnostic capacity and lack of FASD resources across Canada highlights a critical need for continued FASD support. This study identifies gaps in assessment, diagnosis and reporting practices for FASD in children/adolescents across Canada.

Studies evaluating the interactions between Shiga toxin-producing Escherichia coli O157:H7 (O157) and the bovine recto–anal junction (RAJ) have been limited to either in vitro analyses of bacteria, cells, or nucleic acids at the RAJ, providing limited information. Alternatively, expensive in vivo studies in animals have been conducted. Therefore, our objective was to develop a comprehensive in vitro organ culture system of the RAJ (RAJ-IVOC) that accurately represents all cell types present in the RAJ. This system would enable studies that yield results similar to those observed in vivo. Pieces of RAJ tissue, obtained from unrelated cattle necropsies, were assembled and subjected to various tests in order to determine the optimal conditions for assaying bacterial adherence in a viable IVOC. O157 strain EDL933 and E. coli K12 with known adherence differences were used to standardize the RAJ-IVOC adherence assay. Tissue integrity was assessed using cell viability, structural cell markers, and histopathology, while the adherence of bacteria was evaluated via microscopy and culture methods. DNA fingerprinting verified the recovered bacteria against the inoculum. When the RAJ-IVOC was assembled in Dulbecco’s Modified Eagle Medium, maintained at a temperature of 39 °C with 5% CO2 and gentle shaking for a duration of 3–4 h, it successfully preserved tissue integrity and reproduced the expected adherence phenotype of the bacteria being tested. The RAJ-IVOC model system provides a convenient method to pre-screen multiple bacteria-RAJ interactions prior to in vivo experiments, thereby reducing animal usage.

Improper storage conditions or processing of milk leads to potential spoilage and illness, due in part to temperature abuse, allowing bacteria present to grow and spoil the product. However, certain proteins naturally found in raw milk, such as lactoferrin, have reported antibacterial properties. The levels of lactoferrin required to effectively inhibit growth of pathogens have not been investigated thoroughly. This study aimed to examine various concentrations of lactoferrin as a potential biopreservative and as an antimicrobial against the common dairy pathogens Salmonella enterica and Escherichia coli O157:H7. Minimum inhibitory concentration assays were conducted on raw bovine milk in which the bacteria were exposed to varying concentrations of lactoferrin. In the raw milk system, the growth of E. coli O157:H7 was significantly decreased at levels greater than 14.05 mg/mL lactoferrin based on the reduction of tetrazolium salts. For S. enterica, only lactoferrin concentrations at or above 112.5 mg/mL in the milk resulted in reduced growth. Taken together, these results indicate that lactoferrin may have biopreservative potential. To fully examine the practicality and effectiveness of lactoferrin as an antimicrobial additive, a similar study should be conducted using additional (gram-positive) pathogens, such as Bacillus cereus and Listeria monocytogenes. If effective, lactoferrin could prolong the shelf life of dairy products and help reduce the incidence of foodborne illnesses in developing countries with limited refrigeration capability.

Abstract Anthracyclines are highly effective chemotherapeutic agents; however, their clinical utility is limited by severe anthracycline-induced cardiotoxicity (ACT). Genome-wide association studies (GWAS) have uncovered several genetic variants associated with ACT, but the impact of these findings requires further elucidation. We conducted a transcriptome-wide association study (TWAS) using our previous GWAS summary statistics (n = 280 patients) to identify gene expression-related associations with ACT. We identified a genetic association between decreased expression of GDF5 and ACT (Z-score = −4.30, P = 1.70 × 10−5), which was replicated in an independent cohort (n = 845 patients, P = 3.54 × 10−3). Additionally, cell viability of GDF5-silenced human cardiac myocytes was significantly decreased in response to anthracycline treatment. Subsequent gene set enrichment and pathway analyses of the TWAS data revealed that genes essential for survival, cardioprotection and response to anthracyclines, as well as genes involved in ribosomal, spliceosomal and cardiomyopathy pathways are important for the development of ACT.

Alzheimer’s disease (AD) is an age-related neurodegenerative disease that affects 50 million people worldwide, with 10 million new cases occurring each year. The emotional and economic impacts of AD on patients and families are devastating. Approved treatments confer modest improvement in symptoms, and recently one treatment obtained accelerated approval from the United States Food and Drug Administration (FDA) and may have modest disease modifying benefit. Research over the past three decades has established a clear causal linkage between AD and elevated brain levels of amyloid β (Aβ) peptide, and substantial evidence now implicates soluble, non-fibrillar Aβ oligomers (AβOs) as the molecular assemblies directly responsible for AD-associated memory and cognitive failure and accompanying progressive neurodegeneration. The widely recognized linkage of elevated Aβ and AD spawned a comprehensive 20-year therapeutic campaign that focused primarily on two strategies – inhibition of the secretase enzymes responsible for Aβ production and clearance of Aβ peptide or amyloid plaques with Aβ-directed immunotherapeutics. Unfortunately, all clinical trials of secretase inhibitors were unsuccessful. Of the completed phase 3 immunotherapy programs, bapineuzumab (targeting amyloid plaque) and solanezumab (targeting Aβ monomers) were negative, and the crenezumab program (targeting Aβ monomers and to a small extent oligomers) was stopped for futility. Aducanumab (targeting amyloid plaques), which recently received FDA accelerated approval, had one positive and one negative phase 3 trial. More than 25 negative randomized clinical trials (RCTs) have evaluated Aβ-targeting therapeutics, yet none has directly evaluated whether selective blockage of disease-relevant AβOs can stop or reverse AD-associated cognitive decline. Here, we briefly summarize studies that establish the AD therapeutic rationale to target AβOs selectively, and we describe ACU193, the first AβO-selective immunotherapeutic to enter human clinical trials and the first positioned to test the AβO hypothesis of AD.

Abstract Diverse taxa have undergone phenological shifts in response to anthropogenic climate change. While such shifts generally follow predicted patterns, they are not uniform, and interspecific variation may have important ecological consequences. We evaluated relationships among species’ phenological shifts (mean flight date, duration of flight period), ecological traits (larval trophic specialization, larval diet composition, voltinism), and population trends in a butterfly community in Pennsylvania, USA, where the summer growing season has become warmer, wetter, and longer. Data were collected over 7–19 years from 18 species or species groups, including the extremely rare eastern regal fritillary Speyeria idalia idalia. Both the direction and magnitude of phenological change over time was linked to species traits. Polyphagous species advanced and prolonged the duration of their flight period while oligophagous species delayed and shortened theirs. Herb feeders advanced their flight periods while woody feeders delayed theirs. Multivoltine species consistently prolonged flight periods in response to warmer temperatures, while univoltine species were less consistent. Butterflies that shifted to longer flight durations, and those that had polyphagous diets and multivoltine reproductive strategies tended to decline in population. Our results suggest species’ traits shape butterfly phenological responses to climate change, and are linked to important community impacts.

The effectiveness of anthracycline chemotherapeutics (e.g., doxorubicin) is limited by anthracycline-induced cardiotoxicity (ACT). A nonsynonymous variant (S427L) in the retinoic acid receptor-γ (RARG) gene has been associated with ACT. This variant causes reduced RARG activity, which is hypothesized to lead to increased susceptibility to ACT through reduced activation of the retinoic acid pathway. This study explored the effects of activating the retinoic acid pathway using a RAR-agonist, all-trans retinoic acid (ATRA), in human cardiomyocytes and mice treated with doxorubicin. In human cardiomyocytes, ATRA induced the gene expression of RARs (RARG, RARB) and repressed the expression of topoisomerase II enzyme genes (TOP2A, TOP2B), which encode for the molecular targets of anthracyclines and repressed downstream ACT response genes. Importantly, ATRA enhanced cell survival of human cardiomyocytes exposed to doxorubicin. The protective effect of ATRA was also observed in a mouse model (B6C3F1/J) of ACT, in which ATRA treatment improved heart function compared to doxorubicin-only treated mice. Histological analyses of the heart also indicated that ATRA treatment reduced the pathology associated with ACT. These findings provide additional evidence for the retinoic acid pathway's role in ACT and suggest that the RAR activator ATRA can modulate this pathway to reduce ACT.

Previously, we had demonstrated that Escherichia coli O157:H7 (O157) strain 86-24 expresses proteins involved in survival rather than virulence in vitro in rumen fluid from dairy cattle limit fed a maintenance diet. Here, we verified if this observation would be true for different O157 strains grown in vitro in rumen fluid from, and in vivo in the rumen of, animals on contrasting maintenance (high fiber) and lactation (high energy-protein) diets usually limit fed to dairy cattle. For the in vivo studies, an economical, novel, reusable and non-terminal rumen-fistulated animal model permitting simultaneous evaluation of multiple bacterial strains in the bovine rumen was developed. All experiments were conducted in duplicate using different animals to account for host-related variations. The O157 strains included, 86-24, EDL933 and the super shed SS-17. E. coli NalR (#5735), derived from a bovine intestinal commensal E. coli, was included as a control. As expected, diet influenced ruminal pH and volatile fatty acid (VFA) composition. The pH ranged from 6.2-7.0 and total VFA concentrations from 109-141 μM/ml, in animals fed the maintenance diet. In comparison, animals fed the lactation diet had a ruminal pH ranging between 5.18-6.0, and total VFA of 125-219 μM/ml. Strain dependent differences in O157 recovery from the rumen fluid of cattle fed either diet was observed, both in vitro and in vivo, with O157 strains 86-24 and EDL933 demonstrating similar survival patterns. Analysis of the O157 proteomes expressed in the rumen fluid/rumen verified previous observations of adaptive responses. Any difference in the adaptive response was mainly influenced by the animal's diet and growth conditions (in vitro and in vivo) and not the O157 strain. These new insights into the O157 responses could help formulate modalities to control O157 across strains in cattle at all stages of husbandry.

Generational Self-Definition in the Hungarian Theatre in 1980s: Theatrical Functional Trial in Several Performances in Kornis Mihály’s “Kozma” Drama The lecture is about Kornis Mihály’s less known drama with the title “Kozma” directed by Ács János (1986, Kaposvár) and Horvai István (1988, Pesti Színház). For the reconstruction of the play I used the creator’s declarations, photos, criticism, play recordings. Kornis’s third drama takes place in the 1980’s at Lake Balaton on the beach of a closed holiday resort, where three old women enjoy themselves in the heat, when on the effect of a mysterious person named Kozma “coming from above” they begin to confess their secrets. The women’s relatives were men working for the government. Their confessions and their lives are closely related to the previous three decades’ historical events. A scene becomes a common place for the one -dimensional peaceful phenomena and its victims tensioned world. The ones suffering innocently die “in one minute” round the three women who seem to belong to the privileged stratum. The play ends with wild dancing, which can be interpreted as reconciliation or a penalty dance. The drama “Kozma” had a summarizing function in the contemporary dramaturgy. In a reference or partly contains every characteristic and the important historical events of the age presented in the play (1956, 1968 Prague). It can be regarded as a national self-definition, the common experience of the Hungarian playwrights. It should be mentioned the effect of Pilinszky’s “Síremlék” (“Tombstone”) -dramaturgy, which in Hungarian literature created in a unique way the silence of the postwar anxiety (“Gyerekek és katonák”, “Kids and Soldiers”, Pécs, 1981). The premiere in Kaposvár bonus that Ács and Kornis belonged to the same generation and they aimed a valid theatrical presentation without any political tendency. Ács and Horvai directed in a stylized, sensitive way the different layers of the play that is not without mistakes. One of the main strengths of Kozma is the uncertainty in action, their interpretation and the dialogues. Kornis leaves the reason of actions in shadow: the events are questioned. Characters of Kozma do not realize their fate, their mistakes and the fact that they are victims; they hold their burdens alone. This solitude is expressed on the stage with spectacular pantomime and silences. The author emphasizes the tragic situation with the emphasized lack of it. Kornis thought that the beach resembling a lager is “a metaphor with big endurance” that can authenticate every happening. The old women’s naked sunbathing is a metaphor as well, the metaphor of coziness, the “consciousness of possessing”. The human load of the truck is another metaphor. One of the main questions of the play was the problem of nudity on stage which was a problem of the age. (Characters of the women was characterized by the color and pattern of their swimming suits.) The stage-designer decided to solve the problem by signaling. The reality of the abstract space was given by things used on the shores. The background or the foreground was activated with the means of proper changes and movements – during the activity of one of these the other side’s characters were present as lifeless nature. It is an important contribution to the evaluation of the performance that the social atmosphere around the premiere in 1986 in Kaposvár and the one in Budapest in 1988 changed as well, the play rewritten to the second premiere became a play with much clearer structure and the premiere in Budapest could show better the symbolic-metaphoric layers of the drama.

Abstract Background Data are lacking on type of e-liquid vaped among e-cigarette users. Further, few studies assess all sources of nicotine used by e-cigarette users to assess whether poly-nicotine use relates to nicotine dependence (ND). The objectives were to describe young adult e-cigarette users by: (i) type of e-liquid vaped; (ii) poly-nicotine use; (iii) ND symptoms; and (iv) attempts to quit conventional cigarettes. Methods Data were available in cycle 23 of a longitudinal investigation on the natural course of cigarette smoking and ND. A total of 775 young adults (44% male; mean (SD) age 30.5(1.0)) completed mailed self-report questionnaires in 2017–20. Results Of 775 participants, 149 (19.2%) reported past-year e-cigarette use. Overall, 55.0% of e-cigarette users had used cannabis-containing e-liquid (31.5% vaped cannabis e-liquid exclusively); 50.4% used nicotine-containing e-liquid (23.5% vaped nicotine e-liquid exclusively); and 39.9% used e-liquid without nicotine (8.7% vaped e-liquid without nicotine exclusively). Most e-cigarette users (82.6%) used other nicotine-containing products including conventional cigarettes (72.5%); 60.8% reported ND symptoms, rising to 79.4% among those who vaped nicotine-containing e-liquid. Finally, 29.0% tried to quit conventional cigarettes using e-cigarettes in the past-year, but only 16.7% found them helpful. Conclusions E-cigarettes now appear to appeal to a broader market than smokers who want to quit. More than half of young adult e-cigarette users vaped cannabis-containing e-liquid in the past year while only one-quarter had used e-cigarettes to assist with cessation. Most e-cigarette users used multiple nicotine-containing substances (including combustible cigarettes) which were associated with reports of ND symptoms.

Abstract Amyotrophic lateral sclerosis (ALS) is a degenerative motor neuron disease that has been linked to defective DNA repair. Many familial ALS patients harbor autosomal dominant mutations in the gene encoding the RNA/DNA binding protein ‘fused in sarcoma’ (FUS) commonly inducing its cytoplasmic mislocalization. Recent reports from our group and others demonstrate a role of FUS in maintaining genome integrity and the DNA damage response (DDR). FUS interacts with many DDR proteins and may regulate their recruitment at damage sites. Given the role of FUS in RNA transactions, here we explore whether FUS also regulates the expression of DDR factors. We performed RT2 PCR arrays for DNA repair and DDR signaling pathways in CRISPR/Cas9 FUS knockout (KO) and shRNA mediated FUS knockdown (KD) cells, which revealed significant (> 2-fold) downregulation of BRCA1, DNA ligase 4, MSH complex and RAD23B. Importantly, similar perturbations in these factors were also consistent in motor neurons differentiated from an ALS patient-derived induced pluripotent stem cell (iPSC) line with a FUS-P525L mutation, as well as in postmortem spinal cord tissue of sporadic ALS patients with FUS pathology. BRCA1 depletion has been linked to neuronal DNA double-strand breaks (DSBs) accumulation and cognitive defects. The ubiquitin receptor RAD23 functions both in nucleotide excision repair and proteasomal protein clearance pathway and is thus linked to neurodegeneration. Together, our study suggests that the FUS pathology perturbs DDR signaling via both its direct role and the effect on the expression of DDR genes. This underscors an intricate connections between FUS, genome instability, and neurodegeneration.

Abstract Background Organelle genome studies of Fabaceae, an economically and ecologically important plant family, have been biased towards the plastid genome (plastome). Thus far, less than 15 mitochondrial genome (mitogenome) sequences of Fabaceae have been published, all but four of which belong to the subfamily Papilionoideae, limiting the understanding of size variation and content across the family. To address this, four mitogenomes were sequenced and assembled from three different subfamilies (Cercidoideae, Detarioideae and Caesalpinioideae). Results Phylogenetic analysis based on shared mitochondrial protein coding regions produced a fully resolved and well-supported phylogeny that was completely congruent with the plastome tree. Comparative analyses suggest that two kinds of mitogenome expansions have occurred in Fabaceae. Size expansion of four genera (Tamarindus, Libidibia, Haematoxylum, and Leucaena) in two subfamilies (Detarioideae and Caesalpinioideae) occurred in relatively deep nodes, and was mainly caused by intercellular gene transfer and/or interspecific horizontal gene transfer (HGT). The second, more recent expansion occurred in the Papilionoideae as a result of duplication of native mitochondrial sequences. Family-wide gene content analysis revealed 11 gene losses, four (rps2, 7, 11 and 13) of which occurred in the ancestor of Fabaceae. Losses of the remaining seven genes (cox2, rpl2, rpl10, rps1, rps19, sdh3, sdh4) were restricted to specific lineages or occurred independently in different clades. Introns of three genes (cox2, ccmFc and rps10) showed extensive lineage-specific length variation due to large sequence insertions and deletions. Shared DNA analysis among Fabaceae mitogenomes demonstrated a substantial decay of intergenic spacers and provided further insight into HGT between the mimosoid clade of Caesalpinioideae and the holoparasitic Lophophytum (Balanophoraceae). Conclusion This study represents the most exhaustive analysis of Fabaceae mitogenomes so far, and extends the understanding the dynamic variation in size and gene/intron content. The four newly sequenced mitogenomes reported here expands the phylogenetic coverage to four subfamilies. The family has experienced multiple mitogenome size fluctuations in both ancient and recent times. The causes of these size variations are distinct in different lineages. Fabaceae mitogenomes experienced extensive size fluctuation by recruitment of exogenous DNA and duplication of native mitochondrial DNA.

Background:. Morel-Lavallée lesions have been described in the literature since the mid-19th century, yet contemporary clinical management continues to be challenging and remains variable. These closed degloving injuries are usually seen in the setting of blunt trauma and are due to shearing forces, creating a space for the collection of hemolymphatic fluid. These fluid collections can be persistent despite conservative treatment attempts and can require staged surgical interventions. Methods:. A literature search of peer-reviewed articles pertaining to Morel-Lavallée lesions was performed. Articles relating to the pathophysiology, clinical presentation, diagnosis, treatment, and management of complications of Morel-Lavallée lesions were included to formulate recommendations for clinical management. Results:. Fifty-six relevant articles were included in the review. Recognition of these lesions may be delayed and the use of imaging is important in diagnosis. Initial attempts at conservative management may be appropriate for smaller, uncomplicated Morel-Lavallée lesions. Surgical interventions such as open debridement, techniques to close dead space, negative pressure wound therapy, and skin grafts or flaps are critical to managing complex or recurring lesions. An algorithm for the treatment of Morel-Lavallée lesions is proposed based on review of the literature. Conclusions:. Plastic and reconstructive surgeons are frequently involved in more complex Morel-Lavallée lesions such as those exhibiting delayed healing or super-infection. The skillset needed for successful management of patients with Morel-Lavallée lesions is well within the armamentarium of plastic and reconstructive surgeons.

The feasibility of using dwarf kiwi fruits (Actinia arguta Miq.) as a healthy and sustainable food, compared to other types of commercial kiwi fruits, was evaluated in the present study. The overall antioxidant capacity of these fruits was assessed by either extraction-dependent methods (ABTS, ORAC) or the direct method called Quick, Easy, New, CHEap, Reproducible (QUENCHER) (DPPH, FRAP, Folin–Ciocalteu), applied for the first time to analyze kiwi fruits. With this methodology, all the molecules with antioxidant capacity are measured together in a single step, even those with high molecular weight or poor solubility in aqueous extraction systems, such as antioxidant dietary fiber. The effect of kiwi extracts on physiological and induced intracellular reactive oxygen species (ROS) production on IEC-6 cells was also analyzed, as well as total phenolic content (TPC) by Fast Blue BB, flavonols, hydroxycinnamic acids, and hydroxybenzoic acids. A. arguta fruits showed the highest values in all the antioxidant assays, being remarkably higher than the other kiwi species for Q-FRAP and Q-DPPH. Dwarf kiwi showed the highest potential in reducing physiological ROS and the highest values of TPC (54.57 mgGAE/g), being hydroxybenzoic acids the main phenolic family found (2.40 mgGAE/g). Therefore, dwarf kiwi fruits are a natural source of antioxidants compared to conventional kiwi fruits, being a sustainable and healthy alternative to diversify fruits in the diet.

Background Diabetes mellitus (DM) is associated with left ventricular remodeling and incident heart failure, but the association between glycated hemoglobin A1c (HbA1c) and subclinical cardiac disease is not established. We aimed to determine the associations between HbA1c and (1) echocardiographic measures of left ventricular structure and function, and (2) cardiovascular biomarkers: cardiac troponin T, NT‐proBNP (N‐terminal pro‐B‐type natriuretic peptide), and CRP (C‐reactive protein). Methods and Results Participants (n=3688) born in 1950 from the population‐based ACE (Akershus Cardiac Examination) 1950 Study were classified as DM (HbA1c≥6.5% or self‐reported DM), pre‐DM (HbA1c 5.7%–6.5%), and no‐DM (HbA1c

Extrusion cooking allows the development of value-added products from pulses, such as gluten-free snacks with added functional properties. The main objective of this study was to evaluate the changes induced by the extrusion process on the carbohydrate fraction (total carbohydrates, soluble sugars and oligosaccharides, dietary fiber, and arabinoxylans) of novel flour formulations based on chickpeas and rice enriched with different dietary fiber sources. Moreover, the influence of the addition of fiber-rich ingredients, such as Fibersol® and passion fruit, on the analyzed compounds was also evaluated. Sucrose was the main soluble sugar found in analyzed formulations, and raffinose was the prevalent oligosaccharide, followed by stachyose. The content of total α-galactosides tended to be higher after extrusion cooking. As a consequence of the extrusion treatment, the content of total and soluble dietary fiber was statistically increased in most of the analyzed samples. In general, no significant changes were observed in total arabinoxylan content as a consequence of the extrusion process, while the content of water-soluble arabinoxylans was significantly increased in extruded formulations. It was observed that the content of total available carbohydrates, stachyose, and water-soluble arabinoxylans were significantly influenced by the addition of passion fruit, Fibersol®, and both. The incorporation of these ingredients in gluten-free formulations based on chickpeas and rice allows one to obtain suitable functional formulations for the development of innovative, gluten-free, extruded snack-type products, which could be an interesting alternative for people with celiac disease.

Foodborne diseases affect an estimated 600 million people worldwide annually, with the majority of these illnesses caused by Norovirus, Vibrio, Listeria, Campylobacter, Salmonella, and Escherichia coli. To elicit infections in humans, bacterial pathogens express a combination of virulence factors and toxins. AB5 toxins are an example of such toxins that can cause various clinical manifestations, including dehydration, diarrhea, kidney damage, hemorrhagic colitis, and hemolytic uremic syndrome (HUS). Treatment of most bacterial foodborne illnesses consists of fluid replacement and antibiotics. However, antibiotics are not recommended for infections caused by Shiga toxin-producing E. coli (STEC) because of the increased risk of HUS development, although there are conflicting views and results in this regard. Lack of effective treatment strategies for STEC infections pose a public health threat during outbreaks; therefore, the debate on antibiotic use for STEC infections could be further explored, along with investigations into antibiotic alternatives. The overall goal of this review is to provide a succinct summary on the mechanisms of action and the pathogenesis of AB5 and related toxins, as expressed by bacterial foodborne pathogens, with a primary focus on Shiga toxins (Stx). The role of Stx in human STEC disease, detection methodologies, and available treatment options are also briefly discussed.

Objective: To describe the natural course of cigarette smoking and nicotine dependence (ND) over 1-year in daily smokers ages 15 to 17 living in different social contexts. Method: Cigarette smoking and ND indicators were measured at baseline and 3- and 12-months thereafter among 95 daily smokers with a total of 123 observations from the Nicotine Dependence in Teens (NDIT) Study in Quebec, Canada, and in 111 daily smokers from the TABagisme chez les ADOlescents (TABADO) Study in Lorraine, France. Results: NDIT and TABADO participants initiated smoking a mean (SD) of 3.9 (1.6) and 3.7 (2.0) years prior to baseline, respectively. Despite baseline differences in age, sex, age at initiation, number of cigarettes smoked per day and social context, 85% of participants in both samples reported cravings and responded “yes” to “felt like you really need a cigarette”. Mean (SD) number of cigarettes smoked per day increased from 9.4 (8.1) to 11.8 (8.0) over 1 year in NDIT (adjusted mean difference (95% CI) = 2.4 (0.8, 3.0)), and from 11.5 (6.5) to 13.5 (6.7) in TABADO (adjusted mean difference (95% CI) = 2.0 (0.8, 3.1)). However, most ND indicators in both samples were stable over time and cessation was infrequent. Conclusion: Despite notable differences across samples, the natural course of cigarettes smoked per day, ND symptoms and cessation was similar, suggestive of an underlying biologic rather than social process. To quit, adolescents who smoke daily will likely need (pharmacologic) intervention to counter the biological mechanisms underpinning ND, as well as complementary strategies targeting the social context such as creating social environments favoring cessation success.

Abstract The application of complex network theory to community ecology has enabled quantification of interactions among large suites of species and clarified patterns of community structure across systems. Past analyses, however, have assumed that ecological networks are temporally static and persistent and spatially homogeneous, which could confound inference if species interactions vary over time and space. To evaluate temporal and spatial variation in mutualistic networks, therefore, we evaluated the consistency of a nectarivory/pollination network across years, by season, and over space. We tracked nectaring interactions among 37 butterfly and 58 flowering plant taxa during an 11‐yr period (2007–2017), across each summer and over a grassland landscape in Pennsylvania, USA. The composition of butterflies, plants, and their interactions varied markedly across years, months, and sites. Despite this compositional variation, one metric of network structure, nestedness, was invariant, with interactions much more nested than random across all years, months, and sites. Together with previous studies, this result suggests ecological interaction networks are generally more nested than expected by chance. Other measures of network structure were more variable, especially over time. Numbers of plants and interactions varied by year, month, and site. Connectance and numbers of butterflies varied annually and seasonally. Temporal variation in specialization was also evident for some species at an annual level and for the community across the season. We further found highly stable species were almost always generalists, while highly specialized species were almost always temporally and spatially variable, with few exceptions. Together, these results suggest communities are comprised of a reliable core of generalist species, accompanied by a changing suite of specialist species that, when participating in the network, primarily interact with the reliable core species. Our finding of nested mutualistic network centered on stable‐generalist species accompanied by a changing suite of sporadic specialists indicates dynamic changes in ecological communities vary with topological position. This finding further suggests that, even as rare species are highly threatened by species invasion, climate change, and other anthropogenic perturbations, network structure may be robust to species loss and compositional change from these perturbations.

Early-life viral infections are responsible for pulmonary exacerbations that can contribute to disease progression in young children with cystic fibrosis (CF). The most common respiratory viruses detected in the CF airway are human rhinoviruses (RV), and augmented airway inflammation in CF has been attributed to dysregulated airway epithelial responses although evidence has been conflicting. Here, we exposed airway epithelial cells from children with and without CF to RV in vitro. Using RNA-Seq, we profiled the transcriptomic differences of CF and non-CF airway epithelial cells at baseline and in response to RV. There were only modest differences between CF and non-CF cells at baseline. In response to RV, there were 1,442 and 896 differentially expressed genes in CF and non-CF airway epithelial cells, respectively. The core antiviral responses in CF and non-CF airway epithelial cells were mediated through interferon signaling although type 1 and 3 interferon signaling, when measured, were reduced in CF airway epithelial cells following viral challenge consistent with previous reports. The transcriptional responses in CF airway epithelial cells were more complex than in non-CF airway epithelial cells with diverse over-represented biological pathways, such as cytokine signaling and metabolic and biosynthetic pathways. Network analysis highlighted that the differentially expressed genes of CF airway epithelial cells' transcriptional responses were highly interconnected and formed a more complex network than observed in non-CF airway epithelial cells. We corroborate observations in fully differentiated air–liquid interface (ALI) cultures, identifying genes involved in IL-1 signaling and mucin glycosylation that are only dysregulated in the CF airway epithelial response to RV infection. These data provide novel insights into the CF airway epithelial cells' responses to RV infection and highlight potential pathways that could be targeted to improve antiviral and anti-inflammatory responses in CF.

CRISPR/Cas9 gene editing technology has taken the scientific community by storm since its development in 2012. First discovered in 1987, CRISPR/Cas systems act as an adaptive immune response in archaea and bacteria that defends against invading bacteriophages and plasmids. CRISPR/Cas9 gene editing technology modifies this immune response to function in eukaryotic cells as a highly specific, RNA-guided complex that can edit almost any genetic target. This technology has applications in all biological fields, including plant pathology. However, examples of its use in forest pathology are essentially nonexistent. The aim of this review is to give researchers a deeper understanding of the native CRISPR/Cas systems and how they were adapted into the CRISPR/Cas9 technology used today in plant pathology—this information is crucial for researchers aiming to use this technology in the pathosystems they study. We review the current applications of CRISPR/Cas9 in plant pathology and propose future directions for research in forest pathosystems where this technology is currently underutilized.

We have created the immunodeficient SRG rat, a Sprague-Dawley Rag2/Il2rg double knockout that lacks mature B cells, T cells, and circulating NK cells. This model has been tested and validated for use in oncology (SRG OncoRat®). The SRG rat demonstrates efficient tumor take rates and growth kinetics with different human cancer cell lines and PDXs. Although multiple immunodeficient rodent strains are available, some important human cancer cell lines exhibit poor tumor growth and high variability in those models. The VCaP prostate cancer model is one such cell line that engrafts unreliably and grows irregularly in existing models but displays over 90% engraftment rate in the SRG rat with uniform growth kinetics. Since rats can support much larger tumors than mice, the SRG rat is an attractive host for PDX establishment. Surgically resected NSCLC tissue from nine patients were implanted in SRG rats, seven of which engrafted and grew for an overall success rate of 78%. These developed into a large tumor volume, over 20,000 mm3 in the first passage, which would provide an ample source of tissue for characterization and/or subsequent passage into NSG mice for drug efficacy studies. Molecular characterization and histological analyses were performed for three PDX lines and showed high concordance between passages 1, 2 and 3 (P1, P2, P3), and the original patient sample. Our data suggest the SRG OncoRat is a valuable tool for establishing PDX banks and thus serves as an alternative to current PDX mouse models hindered by low engraftment rates, slow tumor growth kinetics, and multiple passages to develop adequate tissue banks.

S. mitis is an abundant member of the commensal microbiota of the oral cavity and pharynx, which has the potential to promote systemic infections. By analyzing a collection of S. mitis strains isolated from the oral cavity at commensal states or from systemic infections (blood strains), we established that S. mitis ubiquitously express the surface immunodominant protein, PcsB (also called GbpB), required for binding to sucrose-derived exopolysaccharides (EPS). Immuno dot blot assays with anti-PcsB antibodies and RT-qPCR transcription analyses revealed strain-specific profiles of PcsB production associated with diversity in pcsB transcriptional activities. Additionally, blood strains showed significantly higher levels of PcsB expression compared to commensal isolates. Because Streptococcus mutans co-colonizes S. mitis dental biofilms, and secretes glucosyltransferases (GtfB/C/D) for the synthesis of highly insoluble EPS from sucrose, profiles of S. mitis binding to EPS, biofilm formation and evasion of the complement system were assessed in sucrose-containing BHI medium supplemented or not with filter-sterilized S. mutans culture supernatants. These analyses showed significant S. mitis binding to EPS and biofilm formation in the presence of S. mutans supernatants supplemented with sucrose, compared to BHI or BHI-sucrose medium. In addition, these phenotypes were abolished if strains were grown in culture supernatants of a gtfBCD-defective S. mutans mutant. Importantly, GtfB/C/D-associated phenotypes were enhanced in high PcsB-expressing strains, compared to low PcsB producers. Increased PcsB expression was further correlated with increased resistance to deposition of C3b/iC3b of the complement system after exposure to human serum, when strains were previously grown in the presence of S. mutans supernatants. Finally, analyses of PcsB polymorphisms and bioinformatic prediction of epitopes with significant binding to MHC class II alleles revealed that blood isolates harbor PcsB polymorphisms in its functionally conserved CHAP-domain, suggesting antigenic variation. These findings reveal important roles of PcsB in S. mitis-host interactions under commensal and pathogenic states, highlighting the need for studies to elucidate mechanisms regulating PcsB expression in this species.

Maize-legume intercropping is a fundamental component of mixed farming systems in the mid-hills of Nepal. However, its productivity is constrained by several biophysical and social factors, and limited adoption of proven agricultural innovations. In this study, we assessed the productivity impact of a selection of relevant agricultural innovations and changes in the associated perceptions of farmers through a series of two-year participatory on-farm trials. The evaluated innovations resulted in higher yields as compared to farmers' current practices. The active involvement of farmers enlarged our understanding of underlying decision-making factors to adopt or non-adopt agricultural innovations. Additionally, the in-depth farmer engagement in our onfarm trials positively influenced farmer perceptions of the innovations and their interest to adopt the agricultural innovations. Yet, farmers final decisions to adopt some of the evaluated innovations were limited by a host of factors including labour scarcity, the availability of inputs, and by cultural preferences despite the increased yields. This was particularly true for low and medium resource-endowed farmers. This study shows the importance of active farmer participation and context-specific design of research and development projects aiming for local impact.

Abstract Background Accumulation mode particles (AMP) are formed from engine combustion and make up the inhalable vapour cloud of ambient particulate matter pollution. Their small size facilitates dispersal and subsequent exposure far from their original source, as well as the ability to penetrate alveolar spaces and capillary walls of the lung when inhaled. A significant immuno-stimulatory component of AMP is lipopolysaccharide (LPS), a product of Gram negative bacteria breakdown. As LPS is implicated in the onset and exacerbation of asthma, the presence or absence of LPS in ambient particulate matter (PM) may explain the onset of asthmatic exacerbations to PM exposure. This study aimed to delineate the effects of LPS and AMP on airway inflammation, and potential contribution to airways disease by measuring airway inflammatory responses induced via activation of the LPS cellular receptor, Toll-like receptor 4 (TLR-4). Methods The effects of nebulized AMP, LPS and AMP administered with LPS on lung function, cellular inflammatory infiltrate and cytokine responses were compared between wildtype mice and mice not expressing TLR-4. Results The presence of LPS administered with AMP appeared to drive elevated airway resistance and sensitivity via TLR-4. Augmented TLR4 driven eosinophilia and greater TNF-α responses observed in AMP-LPS treated mice independent of TLR-4 expression, suggests activation of allergic responses by TLR4 and non-TLR4 pathways larger than those induced by LPS administered alone. Treatment with AMP induced macrophage recruitment independent of TLR-4 expression. Conclusions These findings suggest AMP-LPS as a stronger stimulus for allergic inflammation in the airways then LPS alone.

The airway epithelium of children with wheeze is characterized by defective repair that contributes to disease pathobiology. Dysregulation of developmental processes controlled by Notch has been identified in chronic asthma. However, its role in airway epithelial cells of young children with wheeze, particularly during repair, is yet to be determined. We hypothesized that Notch is dysregulated in primary airway epithelial cells (pAEC) of children with wheeze contributing to defective repair. This study investigated transcriptional and protein expression and function of Notch in pAEC isolated from children with and without wheeze. Primary AEC of children with and without wheeze were found to express all known Notch receptors and ligands, although pAEC from children with wheeze expressed significantly lower NOTCH2 (10-fold, p = 0.004) and higher JAG1 (3.5-fold, p = 0.002) mRNA levels. These dysregulations were maintained in vitro and cultures from children with wheeze displayed altered kinetics of both NOTCH2 and JAG1 expression during repair. Following Notch signaling inhibition, pAEC from children without wheeze failed to repair (wound closure rate of 76.9 ± 3.2%). Overexpression of NOTCH2 in pAEC from children with wheeze failed to rescue epithelial repair following wounding. This study illustrates the involvement of the Notch pathway in airway epithelial wound repair in health and disease, where its dysregulation may contribute to asthma development.

Aspergillus is increasingly associated with lung inflammation and mucus plugging in early cystic fibrosis (CF) disease during which conidia burden is low and strains appear to be highly diverse. It is unknown whether clinical Aspergillus strains vary in their capacity to induce epithelial inflammation and mucus production. We tested the hypothesis that individual colonising strains of Aspergillus fumigatus would induce different responses. Ten paediatric CF Aspergillus isolates were compared along with two systemically invasive clinical isolates and an ATCC reference strain. Isolates were first characterised by ITS gene sequencing and screened for antifungal susceptibility. Three clusters (A−C) of Aspergillus isolates were identified by ITS. Antifungal susceptibility was variable, particularly for itraconazole. Submerged CF and non-CF monolayers as well as differentiated primary airway epithelial cell cultures were incubated with conidia for 24 h to allow germination. None of the clinical isolates were found to significantly differ from one another in either IL-6 or IL-8 release or gene expression of secretory mucins. Clinical Aspergillus isolates appear to be largely homogenous in their mucostimulatory and immunostimulatory capacities and, therefore, only the antifungal resistance characteristics are likely to be clinically important.

Given the importance of colour in the general acceptance or rejection of a product, the use of colorants is a widespread practice, particularly in the food industry. At the same time, with the increasing consumers’ awareness of the health effects that some artificial colorants can exert, there is a growing tendency to prioritize foodstuffs containing natural additives. In this work, Morus nigra L. and Rubus fruticosus L. fruit juices were characterized in terms of anthocyanins, organic acids, free sugars, and tocopherols, as also regarding their bioactive properties. Given their richness in anthocyanins, this study also aimed to prepare different solid colouring formulations by the spray-drying technique, using as stabilizers maltodextrin and arabic gum. Six free sugars and two organic acids were detected in the fruit juices, as well as the four tocopherol isoforms. Two cyanidin derivatives were found in M. nigra (cyanidin-3-O-glucoside and cyanidin-O-rhamnoside) and other four in R. fruticosus (cyanidin-O-hexoside, cyanidin-3-O-glucoside, cyanidin-O-pentoside, and cyanidin-3-O-dioxaloilglucoside). The developed colouring formulations revealed a good stability over time, in terms of anthocyanin concentration and colour parameters, and revealed to be safe for consumption, either concerning their low microbial load and lack of cytotoxicity. Thus, they represent a promising natural alternative to the massively used artificial colorants.

The development of new food products obtained by extrusion processing has increased in recent years. Extrusion is used by the food industry to produce a wide variety of food products, such as ready-to-eat foods (e.g., snacks), among others. Pulses have also gained popularity as novel food ingredients in the formulation of a variety of food and food products, due to their high content of macro and micronutrients, and bioactive compounds that improve the nutritional and functional properties of the final food products. In this review, the impact of extrusion variables on proteins, carbohydrates, vitamins, phenolics and antinutritional compounds in pulses and pulse-based formulations are highlighted. Particularly, the impact of the specific mechanical energy. Also, the preservation, increase and/or reduction in those functional compounds, as a consequence of different extrusion processing conditions, are discussed.

The accumulation of amyloid protein aggregates in tissues is the basis for the onset of diseases known as amyloidoses. Intriguingly, many amyloidoses impact the central nervous system (CNS) and usually are devastating diseases. It is increasingly apparent that neurotoxic soluble oligomers formed by amyloidogenic proteins are the primary molecular drivers of these diseases, making them lucrative diagnostic and therapeutic targets. One promising diagnostic/therapeutic strategy has been the development of antibody fragments against amyloid oligomers. Antibody fragments, such as fragment antigen-binding (Fab), scFv (single chain variable fragments), and VHH (heavy chain variable domain or single-domain antibodies) are an alternative to full-length IgGs as diagnostics and therapeutics for a variety of diseases, mainly because of their increased tissue penetration (lower MW compared to IgG), decreased inflammatory potential (lack of Fc domain), and facile production (low structural complexity). Furthermore, through the use of in vitro-based ligand selection, it has been possible to identify antibody fragments presenting marked conformational selectivity. In this review, we summarize significant reports on antibody fragments selective for oligomers associated with prevalent CNS amyloidoses. We discuss promising results obtained using antibody fragments as both diagnostic and therapeutic agents against these diseases. In addition, the use of antibody fragments, particularly scFv and VHH, in the isolation of unique oligomeric assemblies is discussed as a strategy to unravel conformational moieties responsible for neurotoxicity. We envision that advances in this field may lead to the development of novel oligomer-selective antibody fragments with superior selectivity and, hopefully, good clinical outcomes.

Mutations in the cystic fibrosis transmembrane regulator (CFTR) gene can reduce function of the CFTR ion channel activity and impair cellular chloride secretion. The gold standard method to assess CFTR function of ion transport using the Ussing chamber requires a high number of airway epithelial cells grown at air-liquid interface, limiting the application of this method for high throughput screening of potential therapeutic compounds in primary airway epithelial cells (pAECs) featuring less common CFTR mutations. This study assessed an alternative approach, using a small scale halide assay that can be adapted for a personalized high throughput setting to analyze CFTR function of pAEC.Pediatric pAECs derived from children with CF (pAECCF) were established and expanded as monolayer cultures, before seeding into 96-well plates for the halide assay. Cells were then transduced with an adenoviral construct containing yellow fluorescent protein (eYFP) reporter gene, alone or in combination with either wild-type CFTR (WT-CFTR) or p.Phe508del CFTR. Four days post transduction, cells were stimulated with forskolin and genistein, and assessed for quenching of the eYFP signal following injection of iodide solution into the assay media.Data showed that pAECCF can express eYFP at high efficiency following transduction with the eYFP construct. The halide assay was able to discriminate functional restoration of CFTR in pAECCF treated with either WT-CFTR construct or the positive controls syntaxin 8 and B-cell receptor-associated protein 31 shRNAs.The current study demonstrates that the halide assay can be adapted for pediatric pAECCF to evaluate restoration of CFTR function. With the ongoing development of small molecules to modulate the folding and/or activity of various mutated CFTR proteins, this halide assay presents a small-scale personalized screening platform that could assess therapeutic potential of molecules across a broad range of CFTR mutations.

El presente trabajo tuvo como objetivo sintetizar, caracterizar y evaluar la resistencia a la corrosión de recubrimientos híbridos sol-gel base TEOS/MPS, sintetizados sobre la aleación AA2050-T8 (Al-Cu-Li) bajo diferentes condiciones experimentales de polimerización, tales como solvente y tiempo de envejecimiento, así como diferentes relaciones molares de los precursores inorgánico tetraetoxisilano (TEOS) e híbrido 3-metacriloxipropiltrimetoxisilano (MPS). Se monitoreó la evolución de los soles durante el proceso de envejecimiento mediante espectroscopía infrarroja (FT-IR) y ensayos reológicos. Los recubrimientos se obtuvieron mediante dip-coating y fueron caracterizados a través de microscopía electrónica de barrido (MEB-EDX). La evaluación de la resistencia a la corrosión se realizó mediante ensayos de polarización anódica potenciodinámica, en solución aireada 0,1 M de NaCl. Los resultados obtenidos revelaron que, de los tres solventes evaluados, el etanol es el que permite mejor estabilización del precursor híbrido y favorece el proceso de polimerización de los grupos metacrilato del precursor MPS. Por otra parte, la variación de la relación molar TEOS:MPS conlleva a la obtención de recubrimientos más homogéneos y continuos al incrementar la relación del precursor inorgánico. Finalmente, todos los recubrimientos sintetizados mejoraron la resistencia a la corrosión de la aleación AA2050-T8, en especial los sintetizados bajo la relación TEOS:MPS 80:20.