Clean labelling sodium nitrite at pilot scale: In-situ reduction of nitrate from plant sources and its effects on the overall quality and safety of restructured cooked ham.

Growing health and environmental concerns have increased demand for all-natural products, with a focus on clean labelling. Sodium nitrite is the most widely used additive in the meat industry because it imparts the typical cured flavour and colour to meat products and, most importantly, their microbiological safety. However, due to health concerns, the European Commission is proposing revised regulations to reduce nitrate and nitrite levels in meat products. As a result, the meat industry is actively seeking alternatives. This study explored the production of four cooked hams utilising nitrate-rich vegetable sources combined with two different nitrate-reducing commercial food cultures, alongside a control ham prepared with sodium nitrite (150 ppm). Microbiological, physico-chemical (pH, water activity, nitrate and nitrite concentration, lipid profile, lipid oxidation) and sensory (texture and colour profile) characterisation of the products was carried out. Challenge tests for Listeria monocytogenes, Clostridium sporogenes and Clostridium perfringens have been performed to assess the growth of pathogens, if present in the products. Results revealed comparable microbiological and physico-chemical profiles across ham formulations, with minor differences observed in colour parameters for sample C. The sensory analysis showed that for the pilot ham formulations A and D, there were no significant differences in consumer perception compared to the control ham. In the challenge tests, L. monocytogenes levels were similar in both control and tested hams. There were no significant differences in C. sporogenes and C. perfringens counts at any temperature or between test and control samples. These results indicate that this technology has a potential future in the cured meat sector, as regulators mandate the reduction of added synthetic chemicals and consumers seek healthier and more natural ingredients in their daily diets.

A Self-Reported Study on Explanatory Variables of Stress in Multiple Sclerosis Patients: Exploring the Effect of Physical Conditions and Emotion Regulation Processes.

BACKGROUND: Multiple sclerosis (MS)-linked stress is frequent, multidetermined and facilitates the onset/exacerbation of MS. However, few explanatory models of stress analysed the joint explanatory effect of emotion regulation and clinical outcomes of MS in those patients. OBJECTIVE: This study explored whether self-reported MS-related conditions (number of relapses, fatigue and global disability) and specific emotion regulation processes (experiential avoidance and self-compassion) explain stress symptoms in MS patients. METHODS: The MS sample comprised 101 patients with MS diagnosis receiving treatment in hospitals and recruited through the Portuguese MS Society. The no-MS sample included 134 age-, sex- and years of education-matched adults without MS recruited from the general Portuguese population. Both samples did not report other neurological disorders. Data were collected using self-response measures. RESULTS: All potential explanatory variables differed significantly between samples, with higher scores found in MS patients. In MS clinical sample, those variables and years of education correlated with stress symptoms and predicted stress symptoms in simple linear regression models. These results allowed their selection as covariates in a multiple linear regression model. Years of education, the number of relapses, fatigue and experiential avoidance significantly predicted 51% of stress symptoms' total variance. CONCLUSIONS: This study provides preliminary evidence on the importance of clinicians and researchers considering the simultaneous contribution of years of education, the number of perceived relapses, fatigue and experiential avoidance as factors that can increase vulnerability to stress in MS patients. Psychological intervention programmes that tackle these factors and associated stress symptomatology should be implemented.

Generation of induced pluripotent stem cells from an individual with early onset and severe hypertrophic cardiomyopathy linked to MYBPC3: c.772G > A mutation.

Hypertrophic cardiomyopathy (HCM) is frequently caused by mutations in the MYPBC3 gene, which encodes the cardiac myosin-binding protein C (cMyBP-C). Most pathogenic variants in MYPBC3 are either nonsense mutations or result in frameshifts, suggesting that the primary disease mechanism involves reduced functional cMyBP-C protein levels within sarcomeres. However, a subset of MYPBC3 variants are missense mutations, and the molecular mechanisms underlying their pathogenicity remain elusive. Upon in vitro differentiation into cardiomyocytes, induced pluripotent stem cells (iPSCs) derived from HCM patients represent a valuable resource for disease modeling. In this study, we generated two iPSC lines from peripheral blood mononuclear cells (PBMCs) of a female with early onset and severe HCM linked to the MYBPC3: c.772G > A variant. Although this variant was initially classified as a missense mutation, recent studies indicate that it interferes with splicing and results in a frameshift. The generated iPSC lines exhibit a normal karyotype and display hallmark characteristics of pluripotency, including the ability to undergo trilineage differentiation. These novel iPSCs expand the existing repertoire of MYPBC3-mutated cell lines, broadening the spectrum of resources for exploring how diverse mutations induce HCM. They additionally offer a platform to study potential secondary genetic elements contributing to the pronounced disease severity observed in this individual.

Site-specific dataset of mining and metallurgical residues for resource management.

This geospatial dataset provides a compilation of findings from an evidence-based review of site-specific resource assessments of mining and metallurgical residues. Information pertaining to location, target material, geological knowledge, extractability, resource classification and stakeholder perspectives was collected from publicly available reports, articles, academic theses, and databases. The dataset includes 44 relevant data attributes from 64 mining and metallurgical sites in 27 countries. Resource classification is available for 38 sites. The dataset can be used by evaluators of recovery projects, authorities that provide permits, as well as by decision makers in support of developing regulatory policies. The dataset facilitates future addition of sites by the research community and can be further used as a starting point to bridge the estimates on recoverable quantities to the United Nations Framework Classification (UNFC). The UNFC is a universally applicable scheme for the sustainable management of all energy, primary and secondary mineral resources. Its use is stimulated by the European Commission and is intended to be adopted by geological surveys to harmonize the data on the availability of primary and secondary raw materials in Europe in future.

Generation of induced pluripotent stem cell lines from two unrelated individuals with familial hypertrophic cardiomyopathy carrying MYBPC3 nonsense mutations.

Familial hypertrophic cardiomyopathy (HCM) stands as a predominant heart condition, characterised by left ventricle hypertrophy in the absence of any associated loading conditions, with affected individuals having an increased risk of developing heart failure and sudden cardiac death (SCD). Two induced pluripotent stem cell (iPSC) lines were derived from peripheral blood mononuclear cells obtained from two unrelated individuals with previously reported nonsense mutations in the MYBPC3 gene. The first individual is a 48-year-old male (F26) with the MYBPC3 c.1731G > A HCM mutation, whereas the second individual is a 43-year-old female (F82) carrying the MYBPC3 c.2670G > A HCM mutation. The generated iPSCs exhibit appropriate expression of pluripotency markers, trilineage differentiation capacity and a normal karyotype. This resource contributes to gaining deeper insights into the pathophysiological mechanisms that underlie HCM.

White spot lesions: diagnosis and treatment - a systematic review.

BACKGROUND: White spot lesions represent the first stage of caries and their prevalence has been increasing in recent years, particularly in patients undergoing orthodontic treatment. DIferential diagnosis and lesion activity are essential to decide on the clinical approaches to treatment. The aim of this study is to understand if the new diagnostic tools such as fluorescence, microradiography and computed microtomography have the potential to change the conventional treatment of white spots". METHODS: A systematic search of available studies in the literature was carried out, using PRISMA guidelines, in Pubmed and Scopus electronic databases and manually to identify relevant articles to answer the PICO question: "Do the new diagnostic tools have the potential to change the conventional treatment of white spots?". This systematic review included randomized controlled trials (RCT), cross-sectional and longitudinal studies complying with the following inclusion criteria: (i) studies in humans, (ii) studies about white spot lesions, (iii) studies published between 2012 and 2023, (iv) studies having both diagnosis and treatment and (v) studies with full text available. In this review we excluded other systematic reviews of clinical trials and in vitro studies. The RoB tool was used to assess the risk of bias. RESULTS: The systematic literature search identified 143 potentially relevant references, which after applying the exclusion criteria, resulted in 20 articles. Regarding diagnostic methods, most articles found were based on conventional methods of visual examination (n:10) or fluorescence (n:7). The least referenced diagnostic techniques were based on the use of clinical photographs (n:2), cross-sectional microradiography (n:1) and computed microtomography (n:1). The use of DIAGNOdent was reported by 3 in vitro studies. With regard to therapies, most studies reported the use of infiltrating resin (n:7) and fluoride-based products (n:5). Other studies have reported the use of self-assembling peptide P11-4 (n:1), home care (n:1), casein phosphopeptide-amorphous calcium phosphate (n:2) and hydrochloric acid (n:1). Combination therapies were also considered. CONCLUSION: Diagnostic tool does not have the potential to change the form of treatment, whether it is a conventional method or a more differentiated one.

Assessing Antimicrobial Efficacy on Plastics and Other Non-Porous Surfaces: A Closer Look at Studies Using the ISO 22196:2011 Standard.

The survival and spread of foodborne and nosocomial-associated bacteria through high-touch surfaces or contamination-prone sites, in either healthcare, domestic or food industry settings, are not always prevented by the employment of sanitary hygiene protocols. Antimicrobial surface coatings have emerged as a solution to eradicate pathogenic bacteria and prevent future infections and even outbreaks. Standardised antimicrobial testing methods play a crucial role in validating the effectiveness of these materials and enabling their application in real-life settings, providing reliable results that allow for comparison between antimicrobial surfaces while assuring end-use product safety. This review provides an insight into the studies using ISO 22196, which is considered the gold standard for antimicrobial surface coatings and examines the current state of the art in antimicrobial testing methods. It primarily focuses on identifying pitfalls and how even small variations in methods can lead to different results, affecting the assessment of the antimicrobial activity of a particular product.

Generation of induced pluripotent stem cell lines from two unrelated individuals with familial hypertrophic cardiomyopathy carrying the MYBPC3 missense c.1484G>A mutation.

Familial hypertrophic cardiomyopathy (HCM) is the most common inherited heart condition. HCM patients show left ventricle hypertrophy without any associated loading conditions, being at risk for heart failure and sudden cardiac death. Two induced pluripotent stem cell (iPSC) lines were generated from peripheral blood mononuclear cells obtained from two unrelated individuals, a 54-year-old male (F81) and a 44-year-old female (F93), both carrying the MYBPC3 c.1484G>A HCM mutation. iPSCs show expression of pluripotency markers, trilineage differentiation capacity and a normal karyotype. This resource enables further assessment of the pathophysiological development of HCM.

Uma Escola em Movimento: a experiência da Escola Nacional Paulo Freire / A School on the Move: the experience of the Paulo Freire National School / Una Escuela en Movimiento: la experiencia de la Escuela Nacional Paulo Freire

A finalidade deste artigo é apresentar o trabalho da Escola Nacional Paulo Freire, a partir dos seus antecedentes históricos, dos movimentos que a constroem e objetivos e desafios atuais. O texto também retoma o sentido da homenagem a Paulo Freire e as principais influências do educador no processo político-pedagógico da escola. (AU)

The objective of this work is to present the work of the Escola Nacional Paulo Freire, from its historical antecedents, the movements that build it and current objectives and challenges. The text also takes up the meaning of the homage to Paulo Freire and the main influences of the educator in our political-pedagogical process.(AU)

El objetivo de este trabajo es presentar el trabajo de la Escola Nacional Paulo Freire, desde sus antecedentes históricos, los movimientos que la construyen y los objetivos y desafíos actuales. El texto también retoma el sentido del homenaje a Paulo Freire y las principales influencias del educador en nuestro proceso político-pedagógico. (AU)

Evidence of Australian wild deer exposure to N. caninum infection and potential implications for the maintenance of N. caninum sylvatic cycle.

Infections with the coccidian parasite Neospora caninum affect domestic and wild animals worldwide. In Australia, N. caninum infections cause considerable losses to the cattle industry with seroprevalence of 8.7% in beef and 10.9% in dairy cattle. Conversely, the role of wild animals, in maintaining the parasite cycle is also unclear. It is possible that native or introduced herbivorous species could be reservoir hosts of N. caninum in Australia, but to date, this has not been investigated. We report here the first large-scale screening of N. caninum antibodies in Australian wild deer, spanning three species (fallow, red and sambar deer). Consequently, we also assessed two commercial cELISA tests validated for detecting N. caninum in cattle for their ability to detect N. caninum antibodies in serum samples of wild deer. N. caninum antibodies were detected in 3.7% (7/189, 95% CI 1.8 - 7.45) of the wild deer serum samples collected in south-eastern Australia (n = 189), including 97 fallow deer (Dama dama), 14 red deer (Cervus elaphus), and 78 sambar deer (Rusa unicolor). Overall, our study provides the first detection of N. caninum antibodies in wild deer and quantifies deer's potential role in the sylvatic cycle of N. caninum.

Effectiveness and Durability of a Quaternary Ammonium Compounds-Based Surface Coating to Reduce Surface Contamination.

Foodborne diseases are of major concern as they have a significant impact on public health, both socially and economically. The occurrence of cross-contamination of food in household kitchens is a serious threat and the adoption of safe food practices is of paramount importance. This work aimed to study the effectiveness and durability of a commercial quaternary ammonium compound-based surface coating which, according to the manufacturer, retains its antimicrobial activity for 30 days, and is suitable for all types of hard surfaces for the prevention and/or control of cross-contamination. For that, its antimicrobial efficacy, killing contact time and durability on three different surfaces-polyvinyl chloride, glass, and stainless-steel-against three pathogens-Escherichia coli ATCC 25922, Acinetobacter baumannii ESB260 and Listeria monocytogenes Scott A-were tested according to the current antimicrobial treated surfaces efficacy test (ISO22196:2011). The results showed that the antimicrobial coating was effective against all pathogens with a reduction of >5.0 log CFU/cm2 in less than one minute for the three surfaces, but its durability was less than one week on all surfaces cleaned in the usual manner. Additionally, trace amounts (≤0.2 mg/kg) of the antimicrobial coating, which may migrate into food when contacting the surface, did not show cytotoxicity to human colorectal adenocarcinoma cells. The suggested antimicrobial coating has the potential to significantly reduce surface contamination, ensure surface disinfection and reduce the likelihood of cross-contamination in domestic kitchens, although it is less durable than suggested. The use of this technology in household settings is an attractive complement to the existing cleaning protocols and solutions that are already in place.

Immune Response to SARS-CoV-2 Vaccination in Cancer Patients: A Prospective Study.

Introduction Cancer patients on active treatment are at increased risk of developing coronavirus disease 2019 (COVID-19), making effective immunization of the utmost importance. However, the effectiveness of vaccination in this population is still unclear. This study aims to evaluate the response against COVID-19 in a cohort of patients with active cancer under immunosuppressive therapy. Methods This was a prospective, cross-sectional, single-center study that included patients with cancer under immunosuppressive therapy vaccinated against COVID-19 between April and September 2021. Exclusion criteria were: previous known severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, single-dose vaccine or incomplete vaccination scheme. Immunoglobulin G (IgG) anti-SARS-CoV-2 antibody levels were assessed using 35.2 binding antibody units (BAU)/mL as the positive cut-off. Assessments were performed 14-31 days after the first and second dose and three months after the second dose. Results A total of 103 patients were included. The median age was 60 years. Most patients were being treated for gastrointestinal cancer (n=38, 36.9%), breast cancer (n=33, 32%) or head and neck cancer (n=18, 17.5%). At evaluation, 72 patients (69.9%) were being treated with palliative intent. The majority were being treated with chemotherapy (CT) alone (57.3%). At the first assessment, levels of circulating SARS-CoV-2 IgG consistent with seroconversion were present in 49 patients (47.6%). At the time of the second assessment, 91% (n=100) achieved seroconversion. Three months after the second dose, 83% (n=70) maintained levels of circulating SARS-CoV-2 IgG consistent with seroconversion. In this study, no SARS-CoV-2 infection was reported in the study population. Conclusions Our findings suggest that this group of patients had a satisfactory COVID-19 immunization response. Although promising, this study should be replicated on a wider scale in order to validate these findings.

High-throughput analysis of the transcriptional patterns of sexual genes in malaria.

BACKGROUND: Plasmodium falciparum (Pf) is the leading protozoan causing malaria, the most devastating parasitic disease. To ensure transmission, a small subset of Pf parasites differentiate into the sexual forms (gametocytes). Since the abundance of these essential parasitic forms is extremely low within the human host, little is currently known about the molecular regulation of their sexual differentiation, highlighting the need to develop tools to investigate Pf gene expression during this fundamental mechanism. METHODS: We developed a high-throughput quantitative Reverse-Transcription PCR (RT-qPCR) platform to robustly monitor Pf transcriptional patterns, in particular, systematically profiling the transcriptional pattern of a large panel of gametocyte-related genes (GRG). Initially, we evaluated the technical performance of the systematic RT-qPCR platform to ensure it complies with the accepted quality standards for: (i) RNA extraction, (ii) cDNA synthesis and (iii) evaluation of gene expression through RT-qPCR. We then used this approach to monitor alterations in gene expression of a panel of GRG upon treatment with gametocytogenesis regulators. RESULTS: We thoroughly elucidated GRG expression profiles under treatment with the antimalarial drug dihydroartemisinin (DHA) or the metabolite choline over the course of a Pf blood cycle (48 h). We demonstrate that both significantly alter the expression pattern of PfAP2-G, the gametocytogenesis master regulator. However, they also markedly modify the developmental rate of the parasites and thus might bias the mRNA expression. Additionally, we screened the effect of the metabolites lactate and kynurenic acid, abundant in severe malaria, as potential regulators of gametocytogenesis. CONCLUSIONS: Our data demonstrate that the high-throughput RT-qPCR method enables studying the immediate transcriptional response initiating gametocytogenesis of the parasites from a very low volume of malaria-infected RBC samples. The obtained data expand the current knowledge of the initial alterations in mRNA profiles of GRG upon treatment with reported regulators. In addition, using this method emphasizes that asexual parasite stage composition is a crucial element that must be considered when interpreting changes in GRG expression by RT-qPCR, specifically when screening for novel compounds that could regulate Pf sexual differentiation.

Altered gastrointestinal tract structure and microbiome following cerebral malaria infection.

Cerebral malaria (CM) is the most severe form of malaria with the highest mortality rate and can result in life-long neurological deficits and ongoing comorbidities. Factors contributing to severity of infection and development of CM are not fully elucidated. Recent studies have indicated a key role of the gut microbiome in a range of health conditions that affect the brain, but limited microbiome research has been conducted in the context of malaria. To address this knowledge gap, the impact of CM on the gut microbiome was investigated in mice. C57BL/6J mice were infected with Plasmodium berghei ANKA (PbA) parasites and compared to non-infected controls. Microbial DNA from faecal pellets collected daily for 6-days post-infection were extracted, and microbiome comparisons conducted using 16S rRNA profiling. We identified significant differences in the composition of bacterial communities between the infected and the non-infected groups, including a higher abundance of the genera Akkermansia, Alistipes and Alloprevotella in PbA-infected mice. Furthermore, intestinal samples were collected post-cull for morphological analysis. We determined that the caecal weight was significantly lower, and the small intestine was significantly longer in PbA-infected mice than in the non-infected controls. We concluded that changes in microbial community composition were primarily driven by the infection protocol and, to a lesser extent, by the time of infection. Our findings pave the way for a new area of research and novel intervention strategies to modulate the severity of cerebral malaria disease.

Neuropathic pain, cognitive fusion, and alexithymia in patients with multiple sclerosis: Cross-sectional evidence for an explanatory model of anxiety symptoms.

Multiple sclerosis (MS) presents a high prevalence, a marked increase worldwide, and a relevant impact on patients, public health, and society. Anxiety often cooccurs with MS and can contribute to the worsening of MS symptoms. However, knowledge about predictors of anxiety in Patients with MS (PwMS) is scarce. OBJECTIVE: This preliminary study explored a novel model for anxiety symptoms in PwMS, including neuropathic pain (NeP), cognitive fusion (CF), experiential avoidance (EA), and alexithymia as explanatory factors. METHOD: This cross-sectional study integrated two independent convenience samples: 107 PwMS recruited from the Portuguese Society for Multiple Sclerosis and 97 age- and gender-matched participants without the MS diagnosis (no-MS sample) recruited from the Portuguese general population. Self-report questionnaires that measured the constructs included in the model were administered to both groups. RESULTS: PwMS showed significantly higher values regarding anxiety symptoms and their explanatory variables (NeP, CF, EA, alexithymia) in comparison to non-MS participants. In the MS sample, no correlations were found between anxiety symptoms and sociodemographic and clinical characteristics. NeP, CF, and alexithymia showed significant correlations with anxiety symptoms and significantly explained this symptomatology in simple linear regression models. Thus, these variables were retained in the multiple linear regression model and emerged as significant regressors that together explained 38% of the variance in anxious symptomatology in PwMS. CONCLUSIONS: This preliminary study provides novel evidence on NeP and some maladaptive emotion regulation strategies related to EA/psychological inflexibility, as vulnerability to anxiety in PwMS can be considerably increased by CF and alexithymia. Clinical implications were discussed.

Pseudomonadota in the oral cavity: a glimpse into the environment-human nexus.

The phylum Pseudomonadota is amongst the most represented in the environment, with a comparatively lower prevalence in the human oral cavity. The ubiquity of Pseudomonadota and the fact that the oral cavity is the most likely entry portal of bacteria from external sources underlie the need to better understand its occurrence in the interface environment-humans. Yet, the relevance oral Pseudomonadota is largely underexplored in the scientific literature, a gap that this review aims at addressing by making, for the first time, an overview of the diversity and ecology of Pseudomonadota in the oral cavity. The screening of scientific literature and human microbiome databases unveiled 1328 reports of Pseudomonadota in the oral cavity. Most of these belonged to the classes Beta- and Gammaproteobacteria, mainly to the families Neisseriaceae, Campylobacteriaceae, and Pasteurelaceae. Others also regularly reported include genera such as Enterobacter, Klebsiella, Acinetobacter, Escherichia, Burkholderia, or Citrobacter, whose members have high potential to acquire virulence and antibiotic resistance genes. This review provides evidence that clinically relevant environmental Pseudomonadota may colonize humans via oral cavity. The need for further investigation about Pseudomonadota at the environment-oral cavity interface and their role as vectors potentially involved in virulence and antibiotic resistance transmission is demonstrated. KEY POINTS: • Neisseriaceae, Campylobacteriaceae, and Pasteurelaceae are part of the core oral microbiome • Enterobacteriaceae, Acinetobacter, or Burkholderia are frequent in the oral microbiome • Gut dysbiosis may be associated with colonization by ubiquitous oral Pseudomonadota.

Microbiological Safety of Cut Melons Sold in Portuguese Retail Markets: A Pilot Study.

Due to the increasing consciousness of a healthy diet and pursuit of convenience among consumers, the market for fresh fruit is on the rise, and the melon is among the most welcome of fruits for its sensory attributes and nutritional properties. Consumption safety of cut fruit remains an issue of concern that may affect public health. This study aimed to perform the microbiological characterisation of a melon, Cucumis melo L. var. "Piel de Sapo", cut by retailers, wrapped in plastic cling film and kept at room temperature in local fruit shops. In addition, the possible transfer of relevant foodborne pathogens, during slicing, from the peel to the interior of the melon, and bacterial growth, were also evaluated when the melon slices were stored at abusive temperatures for 2 days. In this pilot study, a low number of samples were characterised microbiologically (26 cut melons), and some isolates were identified by 16S rRNA sequencing. No Listeria spp. or Salmonella spp. were detected in any of the samples, while Escherichia coli and Staphylococcus aureus were present in four and six out of twenty-six samples, respectively. Following artificial contamination of melons with cocktails of Salmonella spp., E. coli and Listeria monocytogenes, it was observed that, despite the smaller number of L. monocytogenes recovered, all the pathogens were transferred from the contaminated peels to the interior of the melons. Furthermore, over storage time, significant differences were observed (p < 0.05) between the counts obtained from melon slices immediately after cutting (0 h), and after 24 and 48 h at 20 °C, with an increase of about 4 log CFU/g in all the pathogens. In conclusion, some cut melons classified as microbiologically unacceptable or unsatisfactory are being sold in local fruit shops in the Porto Metropolitan Area, Portugal. Although absent in the samples analysed, Salmonella spp. and L. monocytogenes, if present, can be transferred from the outside to the inside of the fruit by the cutting blade and, if not consumed immediately and stored at abusive temperatures, this ready-to-eat product poses a risk of infection. This pilot study, performed for the first time in Portugal under these conditions, clearly demonstrates the need for education campaigns to alert local sellers and consumers of the risk posed by cut melons.

Nocardia spp. isolation in chronic lung diseases: Are there differences between patients with pulmonary nocardiosis and Nocardia colonization?

AIMS: Chronic lung diseases are a recognized risk factor for Nocardia spp. INFECTION: Nocardia spp. isolation does not inevitably imply disease, and thus colonization must be considered. Here, we aimed to analyse the differences between pulmonary nocardiosis (PN) and Nocardia spp. colonization in patients with chronic lung diseases. METHODS AND RESULTS: A retrospective study of patients with laboratory confirmation of isolation of Nocardia spp. in at least one respiratory sample was performed. Patients with PN and Nocardia spp. colonization were compared. There were 71 patients with Nocardia spp. identification, 64.8% were male, with a mean age of 67.7 ± 11.2 years. All patients had ≥1 pre-existing chronic lung disease, and 19.7% of patients were immunocompromised. PN and Nocardia spp. colonization were considered in 26.8% and 73.2% of patients, respectively. Symptoms and chest CT findings were significantly more frequent in patients with PN (p < 0.001). During follow-up time, 12 (16.9%) patients died, 6 in PN group. Immunosuppression, constitutional symptoms, haematological malignancy and PN diagnosis were associated with significantly shorter survival times, despite only immunosuppression (HR 3.399; 95% CI 1.052-10.989) and PN diagnosis (HR 3.568; 95% CI 1.078-11.910) remained associated with a higher death risk in multivariate analysis. CONCLUSIONS: PN was associated with clinical worsening, more chest CT findings and worse clinical outcomes. SIGNIFICANCE AND IMPACT OF STUDY: Nocardia spp. isolation in chronic lung disease patients is more common than expected and the differentiation between colonization and disease is crucial.

First Evidence of Entamoeba Parasites in Australian Wild Deer and Assessment of Transmission to Cattle.

Australian wild deer populations have significantly expanded in size and distribution in recent decades. Due to their role in pathogen transmission, these deer populations pose a biosecurity risk to the livestock industry. However, little is known about the infection status of wild deer in Australia. The intestinal parasite Entamoeba bovis has been previously detected in farm and wild ruminants worldwide, but its epidemiology and distribution in wild ruminants remain largely unexplored. To investigate this knowledge gap, faecal samples of wild deer and domestic cattle from south-eastern Australia were collected and analysed for the presence of Entamoeba spp. using PCR and phylogenetic analysis of the conserved 18S rRNA gene. E. bovis parasites were detected at high prevalence in cattle and wild deer hosts, and two distinct Entamoeba ribosomal lineages (RLs), RL1 and RL8, were identified in wild deer. Phylogenetic analysis further revealed the existance of a novel Entamoeba species in sambar deer and a novel Entamoeba RL in fallow deer. While we anticipated cross-species transmission of E. bovis between wild deer and cattle, the data generated in this study demonstrated transmission is yet to occur in Australia. Overall, this study has identified novel variants of Entamoeba and constitutes the first report of Entamoeba in fallow deer and sambar deer, expanding the host range of this parasite. Epidemiological investigations and continued surveillance of Entamoeba parasites in farm ruminants and wild animals will be required to evaluate pathogen emergence and transmission to livestock.

Red Blood Cell BCL-xL Is Required for Plasmodium falciparum Survival: Insights into Host-Directed Malaria Therapies.

The development of antimalarial drug resistance is an ongoing problem threatening progress towards the elimination of malaria, and antimalarial treatments are urgently needed for drug-resistant malaria infections. Host-directed therapies (HDT) represent an attractive strategy for the development of new antimalarials with untapped targets and low propensity for resistance. In addition, drug repurposing in the context of HDT can lead to a substantial decrease in the time and resources required to develop novel antimalarials. Host BCL-xL is a target in anti-cancer therapy and is essential for the development of numerous intracellular pathogens. We hypothesised that red blood cell (RBC) BCL-xL is essential for Plasmodium development and tested this hypothesis using six BCL-xL inhibitors, including one FDA-approved compound. All BCL-xL inhibitors tested impaired proliferation of Plasmodium falciparum 3D7 parasites in vitro at low micromolar or sub-micromolar concentrations. Western blot analysis of infected cell fractions and immunofluorescence microscopy assays revealed that host BCL-xL is relocated from the RBC cytoplasm to the vicinity of the parasite upon infection. Further, immunoprecipitation of BCL-xL coupled with mass spectrometry analysis identified that BCL-xL forms unique molecular complexes with human µ-calpain in uninfected RBCs, and with human SHOC2 in infected RBCs. These results provide interesting perspectives for the development of host-directed antimalarial therapies and drug repurposing efforts.