First report of Colletotrichum siamense causing anthracnose in Oenocarpus bacaba in Brazil.

Bacaba (Oenocarpus bacaba Mart.) is a native palm tree from Brazilian Amazon and Cerrado biomes. This tree produces a small, rounded fruit with dark skin and approximately 1.5 mm thick pulp, extensively utilized for palm heart extraction, juices, and jellies (De Cól et al. 2021). However, several diseases can adversely impact fruit yield and quality. During the 2021 growing season, anthracnose symptoms were observed in Bacaba fruits, with a disease incidence of 58% in fruits collected from the Abreulândia (9°37'15″ S, 49°9'3″ W) and Gurupi (12°25'46" S; 49°16'42" W) municipalities in Tocantins state, Brazil. A total of 198 fruits exhibiting anthracnose symptoms, characterized by deep necrotic spots, were collected. In the laboratory, symptomatic fruits had their external surfaces sterilized for 30 seconds in 70% ethanol, 1 min in 1.5% NaOCl, and then rinsed with sterile distilled water. Sterilized pieces of the fruit tissue were transferred to PDA medium and incubated for 7 days at 28 ºC with a 12 h photoperiod. After this period, two isolates were obtained from the colonies and were identified both macroscopically and microscopically as Colletotrichum sp. The colonies grown at PDA showed a white to grey cottony mycelia, with straight and fusiform conidia, ranging from 14.0 to 21.0 (mean value of 15.8 ± 1.8) µm in length and 4.0 to 7.0 (mean value of 5.5 ± 0.7) µm in width, (n = 50). For species identification, the intergenic spacer between DNA lyase, mating-type locus MAT1-2-1 (APN2/MAT-IGS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glutamine synthetase (GS), and ß-tubulin (TUB) loci were amplified and sequenced. Resulting sequences were deposited in GenBank (OR333843, OR333844, OR333845 and OR333846). BLAST analysis of the partial APN2/MAT-IGS (99%), GAPDH (99,48%), GS (99,32%) and TUB (99,48%) sequences showed highly similarity to C. siamense isolates (IIFT223 and CBS130147). Maximum likelihood multilocus analysis placed the isolate UFTC16 within the C. siamense clade with 98% bootstrap support, clearly assigning the isolate to this species. Morphological features were consistent with the description of C. siamense (Prihastuti et al., 2009). Inoculation of Bacaba fruits and seedlings was conducted to confirm pathogenicity. The surface of uninjured Bacaba fruits was inoculated with two drops (20 µL) of conidial suspension (106 conidia mL-1). The same methodology was adopted to placed healthy leaves of 35-day-old seedlings grown in plastic tubes. Two drops of sterile distilled water were inoculated on nonwounded healthy fruits and seedlings as a negative control. The fruits and seedlings were incubated for five days in a controlled chamber at 28 °C, 70-80% humidity and a "12-h photoperiod". The experiment was conducted with five replicates (five fruits and five seedlings inoculated per isolate) and repeated once. Typical symptoms of anthracnose were observed in the fruits and leaves of Bacaba seedlings five days after inoculation. No symptoms were observed in the negative control. The pathogen was reisolated from symptomatic fruits and leaves, showing similar morphological characteristics as the original isolate, fulfilling Koch's postulates. The identification of C. siamense as the causal agent of Bacaba anthracnose helps in the diagnosis and disease control strategies of the disease. Colletotrichum siamense is a cosmopolitan species and easily found in cultivated and non-cultivated species (Batista et al. 2023). However, to the best of our knowledge, this is the first report of C. siamense causing anthracnose on Bacaba.

Assessing Ability for ChatGPT to Answer Total Knee Arthroplasty-Related Questions.

BACKGROUND: Artificial intelligence in the field of orthopaedics has been a topic of increasing interest and opportunity in recent years. Its applications are widespread both for physicians and patients, including use in clinical decision-making, in the operating room, and in research. In this study, we aimed to assess the quality of ChatGPT answers when asked questions related to total knee arthroplasty. METHODS: ChatGPT prompts were created by turning 15 of the American Academy of Orthopaedic Surgeons Clinical Practice Guidelines into questions. An online survey was created, which included screenshots of each prompt and answers to the 15 questions. Surgeons were asked to grade ChatGPT answers from 1 to 5 based on their characteristics: (1) relevance, (2) accuracy, (3) clarity, (4) completeness, (5) evidence-based, and (6) consistency. There were 11 Adult Joint Reconstruction fellowship-trained surgeons who completed the survey. Questions were subclassified based on the subject of the prompt: (1) risk factors, (2) implant/intraoperative, and (3) pain/functional outcomes. The average and standard deviation for all answers, as well as for each subgroup, were calculated. Inter-rater reliability (IRR) was also calculated. RESULTS: All answer characteristics were graded as being above average (ie, a score > 3). Relevance demonstrated the highest scores (4.43 ± 0.77) by surgeons surveyed, and consistency demonstrated the lowest scores (3.54 ± 1.10). ChatGPT prompts in the Risk Factors group demonstrated the best responses, while those in the Pain/Functional Outcome group demonstrated the lowest. The overall IRR was found to be 0.33 (poor reliability), with the highest IRR for relevance (0.43) and the lowest for evidence-based (0.28). CONCLUSIONS: ChatGPT can answer questions regarding well-established clinical guidelines in total knee arthroplasty with above-average accuracy but demonstrates variable reliability. This investigation is the first step in understanding large language model artificial intelligence like ChatGPT and how well they perform in the field of arthroplasty.

Kinetics and mathematical model of killer/sensitive interaction under different physicochemical conditions of must/wine: a study from a biological point of view.

Fermentation of grape must to wine is carried out by a complex microbial mixture, which also involves spoilage yeasts of wine. The latter yeasts produce organoleptic changes that cause significant economic losses to the wine industry. SO2 is traditionally used to control this spoilage populations, but because of its harmful effects on human health, biocontrol has emerged as an alternative treatment. Although studies have been carried out to select biocontroller yeasts and examine their underlying mechanisms of action, reports on their application have not been published yet. To better understand the interaction and the successful application of biocontrol, the use of mathematical models, among other methods, is important, as they facilitate the prediction of success or failure of the antagonist. The objective of the present study was to use an existing mathematical model to obtain information about the yeast's interaction assayed and to validate its predictive use under different physicochemical conditions during the wine fermentation, and eventually predict biocontrol kinetics. The mathematical model was applied to the fermentation conditions and provided information on the kinetic parameters of the biocontrol interaction and allowed interpretations about other parameters. The model was applied in the different physicochemical conditions for the biocontrol and did not fit correctly to experimental data, and therefore an improvement was proposed which was successful and presented new hypotheses.

Occurrence of Colletotrichum musicola causing anthracnose on Manihot esculenta.

Cassava (Manihot esculenta) is one of the main food sources of energy in developing countries owing to its starch-rich roots (Pinweha et al., 2015). Anthracnose is considered the most destructive disease of the aerial part of this crop (Bragança et al., 2016; Liu et al., 2019), and it is caused by species such as Colletotrichum plurivorum, C. karstii, C. fructicola, C. siamense (Liu et al., 2019), and C. theobromicola (Oliveira et al, 2016). In 2019, leaves with irregular necrotic spots, typical symptoms of anthracnose, were collected in Pará, Brazil. Commercial sampled fields showed 20% of incidence of anthracnose. Colletotrichum strains were isolated and cultured on potato dextrose agar at 25 ºC with a 12-h light photoperiod from surface-disinfected (70% alcohol and 1% sodium hypochlorite) lesion transition area. Five of the obtained isolates exhibited brown colonies on the upper and lower surfaces. Conidia were hyaline, cylindrical and aseptate, 12.82-15.23 µm × 3.52-5.25 µm in size. These phenotypic characters were similar to those belonging to C. orchidearum sensu lato (Damm et al. 2019). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), ß-tubulin (TUB2), chitin synthase 1(CHS-1), and histone HIS3 partial gene were amplified and sequenced for one representative isolate (UFT/Coll89). Sequences were deposited in GenBank [Accession numbers: MT396235 (GAPDH), MT800856 (TUB2), MT800870 (CHS-1), and MT856672 (HIS3)]. BLASTn searches of CHS-1 and HIS3 sequences showed 100% identity to C. musicola. Maximum Likelihood Phylogenetic analysis, including previously published sequences of closely related species, placed the isolate from Cassava in the C. musicola clade with 100% support, and confidently it assigned to this species. Pathogenicity was proven with inoculations by spraying a conidial suspension (106 conida mL-1) on 3-month-old cassava plants (three unwounded leaves per plant). The plants were placed in a humid chamber at 25 °C for 48h, and a 12-h photoperiod. The negative control was represented by plants inoculated with sterile distilled water. The experiment was repeated twice. The same symptoms observed in the field were reproduced only in inoculated leaves, from which the pathogen was reisolated lesions fulfilling Koch's postulates. No symptoms were observed on the negative control. To our knowledge, this is the first report of C. musicola joining a group of new and emergent species of Colletotrichum causing anthracnose in cassava producing regions around the world. The identification of this species causing cassava anthracnose is crucial to improve the disease control strategies and resistance breeding programs.

Potential Action Mechanism and Inhibition Efficacy of Morinda citrifolia Essential Oil and Octanoic Acid against Stagonosporopsis cucurbitacearum Infestations.

The use of plant-based products has been shown to efficiently inhibit fungi-mediated diseases in agricultural crops. Here, we extracted and evaluated the composition of noni, Morinda citrifolia L., essential oil and assessed its activities against Stagonosporopsis cucurbitacearum in Cucumis melo L. Using in silico molecular approaches, potential interactions between the essential oil major components and S. cucurbitacearum tyrosine-tRNA ligase were predicted. Finally, we also measured the potential interference of plant physiology (the stomatal conductance and net photosynthesis) mediated by the application of the M. citrifolia essential oil. Chromatographic analysis revealed that octanoic acid (75.8%), hexanoic acid (12.8%), and isobutyl pent-4-enyl carbonate (3.1%) were the major essential oil compounds. Octanoic acid and noni essential oil, when used as preventive measures, reduce fungal mycelial growth at a concentration of 5 mg/mL without causing significant damage to the treated leaves, which reinforces their efficacies as preventive tools against S. cucurbitacearum. Molecular docking analyses predicted very stable interactions between the major essential oil constituents and S. cucurbitacearum tyrosine-tRNA ligase, suggesting the interference of these plant-based molecules upon enzyme activation. Octanoic acid and M. citrifolia essential oil at concentrations of 20 mg/mL decreased the stomatal conductance and net photosynthesis rate of melon plants, resulting in robust phytotoxicity. Collectively, our findings indicated that despite the phytotoxicity risks at higher concentrations, M. citrifolia essential oil and octanoic acid, have potential as alternative tools for the integrative management of S. cucurbitacearum.

Predictors of mortality and clinical characteristics among carbapenem-resistant or carbapenemase-producing Enterobacteriaceae bloodstream infections in Spanish children.

BACKGROUND: Carbapenem-resistant Enterobacteriaceae (CRE) are an emerging problem in the paediatric population worldwide with high mortality rates in bloodstream infection (BSI). OBJECTIVES: To evaluate predictors of 30 day mortality in CRE BSI in a paediatric cohort. METHODS: A retrospective observational single-centre study (December 2005-August 2018) was conducted. Cases of CRE BSI in children 0 to 16 years were included. Microbiological identification (MALDI Biotyper) and antimicrobial susceptibility testing (Vitek2® and MicroScan panel NBC44) according to EUCAST breakpoints were performed. PCR OXVIKP® was used to confirm carbapenemase genes (OXA-48, VIM, KPC, NDM). Demographic characteristics, underlying diseases, source of bacteraemia, antimicrobial therapy and outcomes were collected from medical records. Survival analysis to establish predictors of 30 day mortality was performed. RESULTS: Thirty-eight cases were included; 76.3% were hospital-acquired infections and 23.7% related to healthcare. All patients had at least one underlying comorbidity and 52.6% were recipients of an organ transplant. VIM carbapenemase was the predominant mechanism (92.1%). Previous CRE colonization or infection rate was 52.6%. Intestinal tract (26.3%) and vascular catheter (21.1%) were the most common sources of infection. Crude mortality within 30 days was 18.4% (7/38); directly related 30 day mortality was 10.5%. Conditions associated with an increment in 30 day mortality were intensive care admission and inadequate empirical therapy (P < 0.05). Combination-antibiotic targeted treatment and a low meropenem MIC were not related to improved survival. CONCLUSIONS: CRE BSI mortality rate is high. The most important factor related to 30 day survival in our CRE BSI cohort in children was empirical treatment that included at least one active antibiotic.

First report of Colletotrichum truncatum causing anthracnose in cassava in Brazil.

Cassava (Manihot esculenta Crantz) presents significant economic importance in Brazil and other developing countries due to its use in human and animal feeding. In 2019, cassava plants sampled in Pará state (Brazil) presented necrotic and irregular leaf spots, characteristic symptoms of cassava anthracnose. About 90% of the plants were symptomatic, and disease severity was higher during months with high temperature and humidity. Fragments of symptomatic tissues were removed from the lesion transition area, surface disinfested (45 s in 70% ethanol, 1 min in 1% NaOCl, and rinsed twice in sterile water), and plated on potato dextrose agar. Cultures were incubated at 25 °C under continuous light for 7 days. Among the obtained isolates, seven presented grey felt-like mycelium with white sectors, reverse greyish, and hyaline, aseptate, smooth-walled, falcate conidia with average size 20.7-30.7 (26.1 ± 2.1) × 2.4-4.8 (3.5 ± 0.5) µm. Phenotypical features were similar to C. truncatum (Damm et al. 2019). The representative isolate UFT/Coll87 was chosen for further assays. The identity of the isolate was determined by maximum likelihood analysis using sequences of actin (ACT, GenBank accession number MT321653), ß-tubulin (TUB2, MT856673) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, MT800857) partial regions. Colletotrichum isolate from cassava nested with C. truncatum isolates in a clade with 100% support, being confidently assigned to this species. Koch's postulates were fulfilled to confirm the pathogenicity of UFT/Coll87. Inoculation was carried out in three cassava plants by spraying a conidial suspension (106 conida mL-1) on unwounded leaves (three leaves per plant). Plants sprayed with sterile water represented negative control. Inoculated plants were kept in a humid chamber for 48 h, 25 °C, and a 12-h photoperiod. The experiment was repeated 2 times. Typical cassava anthracnose symptoms were observed 10 days after inoculation. No symptoms were observed in negative control. The pathogen was reisolated from symptomatic leaves and was phenotypically identical to the original isolate UFT/Coll87, fulfilling Koch's postulates. Colletotrichum fructicola, C. karstii, C. plurivorum, and C. siamense were reported causing cassava anthracnose in China (Liu et al. 2019). In Brazil, C. chrysophilum, C. fructicola, C. siamense and C. theobromicola were reported in association with cassava (Bragança et al. 2016; Oliveira et al. 2018; Machado et al. 2020). To our knowledge, this is the first report of C. truncatum causing cassava anthracnose worldwide. Our finding is important for disease management due to the high host range of C. truncatum. The pathogen can reduce the cassava yield, and the crop may serve as a potential inoculum source since it is commonly cultivated near to other crops that are also infected by C. truncatum.

Botanical and chemical fungicides in the treatment of commercial seeds of Brachiaria brizantha and Panicum maximum.

Brazil is the largest producer, consumer, and exporter of forage seeds in the world. However, the high incidence of pathogens threatens the seed production system, resulting in decreases in production and quality. Currently, the emphasis has been placed on the use of alternative products as a way of maintaining productivity and environmental sustainability. Thus, the objective of this study was to evaluate the mycoflora associated with the commercial seeds of Brachiaria brizantha and Panicum maximum treated with chemical and alternative fungicide. To test with chemical and alternative treatments, seeds were subjected to fluazinam + thiophanate-methyl, fludioxonil + metalaxyl-M, noni essential oil, rosemary pepper essential oil, sulfuric acid. There was also an untreated control. The most efficient products in the control of incident fungi were, in decreasing order, fluazinam + thiophanate-methyl, fludioxonil + metalaxyl-M, noni essential oil, rosemary pepper essential oil, and sulfuric acid. Fungicides mainly controlled the genera Alternaria, Aspergillus, Bipolaris, Chaetomium, Cladosporium, Colletotrichum, Curvularia, Fusarium, Penicillium, Phoma and Rhizopus. Essential oils showed positive effects in controlling Bipolaris, Curvularia, and Fusarium. Sulfuric acid reduced the incidence of Bipolaris, Colletotrichum, Curvularia, Fusarium, Phoma, and Rhizopus. Seed treatment is one of the promising strategies for obtaining seeds of a healthy quality. As a result, the fungicides and botanical components listed above were promising in reducing fungi associated with forage seeds and can be used for seed treatments and in the development of new products with a natural base through essential oils.

First report of Colletotrichum chrysophillum causing cassava anthracnose in Brazil.

Cassava (Manihot esculenta Crantz) has significant socioeconomic relevance in Brazil and other developing countries, as one of the main sources of carbohydrates for human and animal consumption (De Oliviera et al., 2011). Among the cassava crop diseases, anthracnose is one of the main limiting factors for production and may be caused by species like Colletotrichum plurivorum, C. karstii, C. fructicola, and C. siamense (Bragança et al., 2016; Liu et al., 2019; Oliveira et al., 2016; Sangpueak; Phansak; Buensanteai, 2018). Severity in the field is variable, depending on the resistance of the variety used and is also highly influenced by the climate, being the most severe disease under high humidity and high temperature. Under these conditions, it can cause losses of up to 100%. In 2019, cassava leaves presenting dark brown necrotic injuries of different sizes and irregular borders-typical anthracnose symptoms- were collected from commercial plantations in the states of Pará and Tocantins, Brazil. Symptomatic tissue fragments were superficially disinfected, placed in plates with potato dextrose agar (PDA), and incubated under 25 ± 2 °C for seven days. In the 56 isolates used in the morphological identification, the colonies were white and gray at the top and dark gray in the bottom with sector formation. The conidia were hyaline, cylindrical, and aseptic, 10.04 to 17.83 µm long × 3.29 to 5.75 µm wide. These phenotypical characteristics were similar to those of C. gloeosporioides lato sensu species (Weir et al., 2012). Genomic DNA was extracted from two representative isolates (UFT/Coll69, collected in the municipality of Casa de Tábua-PA; UFT/Coll82, collected in Pau Darco-PA) and the APN2 / MAT-IGS, DNA lyase (Apn2), and glyceraldehyde-3-phosphate dehydrogenase-IGS (GAP2-IG) intergenic spacers were amplified and sequenced. The nucleotide sequences were deposited in the GenBank (accession numbers: MT409462, MT396231, MT759633, MT396239, MT396232, MT800846). The BLASTn (Basic Local Alignment Search Tool) showed a 99 to 100% similarity with Colletotrichum chrysophillum. The maximum likelihood phylogenetic analysis grouped the isolates in the C. chrysophillum clade, with a high bootstrap value (98%). Based on morphocultural characteristics and the phylogenetic analysis, the isolates associated with M. esculenta anthracnose were identified as C. chrysophillum, with a frequency of 6.67% among Colletotrichum colonies isolated from cassava leaves. The inoculation of three isolates was carried out in three plants, three leaves for each plant, by spraying spore solution with a concentration of 1×106 conidia / ml, without wounding the leaves and placed in a humid chamber at 25 ° C for ten days. Control plants were inoculated with sterile distilled water. From the 2nd day after inoculation, small irregular necrotic lesions appeared that increased in size over time, while control plants remained asymptomatic. Both were pathogenic and the symptoms caused after inoculation were similar to each other and to those observed in the field. In Brazil, anthracnose by C. chrysophillum was reported in cashew (Veloso et al., 2018) and banana trees (Vieira et al., 2017). To our knowledge, this is the first report of cassava anthracnose disease by C. chrysophillum.

[Monitoring of COVID-19 patients by telemedicine with telemonitoring]. / Telemedicina con telemonitorización en el seguimiento de pacientes con COVID-19.

AIM: To asses if telemedicine with telemonitoring is a clinically useful and secure tool in the tracking of patients with COVID-19. METHODS: A prospective observational study of patients with COVID-19 diagnosis by positive PCR considered high-risk tracked with telemedicine and telemonitoring was conducted in the sanitary area of Lugo between March 17th and April 17th, 2020. Two groups of patients were included: Outpatient Tracing from the beginning and after discharge. Every patient sent a daily clinical questionnaire with temperature and oxygen saturation 3 times a day. Proactive monitoring was done by getting in touch with every patient at least 11 a day. RESULTS: 313 patients (52.4% female) with a total average age of 60.9 (DE 15.9) years were included. Additionally, 2 patients refused to join the program. Since the beginning, 224 were traced outpatient and 89 after being discharged. Among the first category, 38 (16.90%) were referred to Emergency department on 43 occasions; 18 were hospitalized (8.03%), and 2 deceased. Neither deaths nor a matter of vital emergency occurred at home. When including patients after admissions monitoring was done in 304 cases. One patient re-entered (0.32%) to the hospital, and another one left the program (0.32%). The average time of monitoring was 11.64 (SD 3.58) days, and 224 (73.68%) patients were discharged during the 30 days of study. CONCLUSIONS: Our study suggests that telemedicine with home telemonitoring, used proactively, allows for monitoring high-risk patients with COVID-19 in a clinically useful and secure way.

PET imaging of dopamine-D2 receptor internalization in schizophrenia.

Recent genetic, molecular and post-mortem studies suggest impaired dopamine (DA)-D2 receptor (D2R) trafficking in patients with schizophrenia (SZ). Imaging and preclinical studies have shown agonist-induced D2R internalization can be imaged with positron emission tomography (PET) using D2R radiotracers combined with psychostimulant challenge. This is feasible if radiotracer binding is measured when postchallenge DA levels have returned to baseline, following the initial competition phase between DA and radiotracer for binding to D2R. Here we used 'late-phase' imaging after challenge to test the hypothesis that impaired D2R internalization in SZ leads to blunted late-phase displacement, or a faster return to baseline, in patients compared with healthy controls (HCs). We imaged 10 patients with SZ and 9 HCs with PET and [11C]raclopride at baseline and two times (3-5 and 6-10 h) following 0.5 mg kg-1 dextroamphetamine. We measured binding potential relative to non-displaceable compartment (BPND) and derived percent reduction from baseline (ΔBPND) for each postamphetamine scan. To test the hypothesis that time course of return of striatal BPND to baseline differed between SZ and HCs, we implemented a linear model with ΔBPND as dependent variable, time after amphetamine as repeated measure and time after amphetamine and diagnostic group as fixed effects. Neither diagnostic group nor interaction of diagnostic group-by-time after amphetamine significantly affected striatal ΔBPND (F=1.38, P=0.26; F=0.51, P=0.61). These results show similar pattern of return of BPND to baseline as a function of time in patients with SZ and HC, suggesting that striatal D2R internalization as measured by our imaging paradigm is normal in patients with SZ.

Plant-expressed Fc-fusion protein tetravalent dengue vaccine with inherent adjuvant properties.

Dengue is a major global disease requiring improved treatment and prevention strategies. The recently licensed Sanofi Pasteur Dengvaxia vaccine does not protect children under the age of nine, and additional vaccine strategies are thus needed to halt this expanding global epidemic. Here, we employed a molecular engineering approach and plant expression to produce a humanized and highly immunogenic poly-immunoglobulin G scaffold (PIGS) fused to the consensus dengue envelope protein III domain (cEDIII). The immunogenicity of this IgG Fc receptor-targeted vaccine candidate was demonstrated in transgenic mice expressing human FcγRI/CD64, by induction of neutralizing antibodies and evidence of cell-mediated immunity. Furthermore, these molecules were able to prime immune cells from human adenoid/tonsillar tissue ex vivo as evidenced by antigen-specific CD4+ and CD8+ T-cell proliferation, IFN-γ and antibody production. The purified polymeric fraction of dengue PIGS (D-PIGS) induced stronger immune activation than the monomeric form, suggesting a more efficient interaction with the low-affinity Fcγ receptors on antigen-presenting cells. These results show that the plant-expressed D-PIGS have the potential for translation towards a safe and easily scalable single antigen-based tetravalent dengue vaccine.

PET imaging of dopamine-D2 receptor internalization in schizophrenia.

This corrects the article DOI: 10.1038/mp.2017.107.

Epidemiology and control measures of an OXA-48-producing Enterobacteriaceae hospital-wide oligoclonal outbreak.

The main objective of our study was to describe the epidemiological and microbiological features of an oligoclonal hospital-wide outbreak caused by OXA-48-producing Enterobacteriaceae (OXA-48-PE). OXA-48 is a carbapenemase belonging to Ambler class D beta-lactamases, identified frequently in the Mediterranean and Southern European countries, and associated with several Enterobacteriaceae species. An outbreak of OXA-48-PE with a complex epidemic pattern was detected in January 2011. Initial control measures included contact precautions and the reinforcement of infection control practices, but despite all efforts made, the epidemiological situation hardly changed and new measures were implemented during 2013. An observational retrospective study was performed to describe the main features of the outbreak and to analyse the cumulative incidence (CI) trends. Eight hundred and 16 patients colonised or infected by OXA-48-PE were identified during the 2-year period (January 2013-December 2014), female 46%, mean age (s.d.), 71.6 (15.2). The samples isolated in the incident cases were rectal swabs (80%), urine samples (10.7%), blood samples (2.8%) and other clinical samples (6.6%). The most frequent OXA-48-PE was Klebsiella pneumoniae. Eleven different clones were identified, but K. pneumoniae sequence types 11 and 405 were predominant: ST11 (64.2%) and ST405 (29.3%). OXA-48-PE CI trend suffered a statistically significant change in August 2013, which continued the following months. Though we could not eradicate the outbreak, we observed a statistically significant drop in CI after an intervention for OXA-48-PE control, based on patient cohort, active surveillance, electronic alerts and reinforcement of infection control measures in a tertiary hospital.

Murine IL-4Δ2 splice variant down-regulates IL-4 activities independently of IL-4Rα binding and STAT-6 phosphorylation.

IL-4 is a pleiotropic cytokine that is highly Th2 polarizing. The ratio of IL-4 and its splice variant IL-4Δ2 observed in human health and disease suggests a role for both isoforms. In the present study, the biological function of murine IL-4Δ2 and the potential mechanism of action were studied. We report for the first time the generation of a functional, recombinant murine IL-4Δ2 form which is suggestive of its possible biological role in this species. Recombinant murine IL-4Δ2 inhibited IL-4 mediated cellular processes in macrophages and lymphocytes. Specifically, (i) it reversed IL-4 mediated inhibition of IFN-γ induced nitric oxide release by macrophages, (ii) inhibited IL-4 mediated induction of T cell proliferation, and (iii) prevented IL-4 stimulation of IgE synthesis by B cells. However, IL-4Δ2 did not compete with IL-4 for IL-4Rα binding and did not interfere with the downstream STAT-6 phosphorylation in T cells, suggesting an alternative mechanism for its antagonism of specific IL4-driven effects. These findings suggest that the mouse is a suitable experimental model for studies of the biology of IL-4 and its alternative splice variant.

Discontinuation of contact precautions with the introduction of universal daily chlorhexidine bathing.

Contact precautions are a traditional strategy to prevent transmission of methicillin-resistant Staphylococcus aureus (MRSA). Chlorhexidine bathing is increasingly used to decrease MRSA burden and transmission in intensive care units (ICUs). We sought to evaluate a hospital policy change from routine contact precautions for MRSA compared with universal chlorhexidine bathing, without contact precautions. We measured new MRSA acquisition in ICU patients and surveyed for MRSA environmental contamination in common areas and non-MRSA patient rooms before and after the policy change. During the baseline and chlorhexidine bathing periods, the number of patients (453 vs. 417), ICU days (1999 vs. 1703) and MRSA days/1000 ICU days (109 vs. 102) were similar. MRSA acquisition (2/453 vs. 2/457, P = 0·93) and environmental MRSA contamination (9/474 vs. 7/500, P = 0·53) were not significantly different between time periods. There were 58% fewer contact precaution days in the ICU during the chlorhexidine period (241/1993 vs. 102/1730, P < 0·01). We found no evidence that discontinuation of contact precautions for patients with MRSA in conjunction with adoption of daily chlorhexidine bathing in ICUs is associated with increased MRSA acquisition among ICU patients or increased MRSA contamination of ICU fomites. Although underpowered, our findings suggest this strategy, which has the potential to reduce costs and improve patient safety, should be assessed in similar but larger studies.

Cardiac arrhythmia and late-onset muscle weakness caused by a myofibrillar myopathy with unusual histopathological features due to a novel missense mutation in FLNC.

Myofibrillar myopathies (MFM) are mostly adult-onset diseases characterized by progressive morphological alterations of the muscle fibers beginning in the Z-disk and the presence of protein aggregates in the sarcoplasm. They are mostly caused by mutations in different genes that encode Z-disk proteins, including DES, CRYAB, LDB3, MYOT, FLNC and BAG3. A large family of French origin, presenting an autosomal dominant pattern, characterized by cardiac arrhythmia associated to late-onset muscle weakness, was evaluated to clarify clinical, morphological and genetic diagnosis. Muscle weakness began during adult life (over 30 years of age), and had a proximal distribution. Histology showed clear signs of a myofibrillar myopathy, but with unusual, large inclusions. Subsequently, genetic testing was performed in MFM genes available for screening at the time of clinical/histological diagnosis, and desmin (DES), αB-crystallin (CRYAB), myotilin (MYOT) and ZASP (LDB3), were excluded. LMNA gene screening found the p.R296C variant which did not co-segregate with the disease. Genome wide scan revealed linkage to 7q.32, containing the FLNC gene. FLNC direct sequencing revealed a heterozygous c.3646T>A p.Tyr1216Asn change, co-segregating with the disease, in a highly conserved amino acid of the protein. Normal filamin C levels were detected by Western-blot analysis in patient muscle biopsies and expression of the mutant protein in NIH3T3 showed filamin C aggregates. This is an original FLNC mutation in a MFM family with an atypical clinical and histopathological presentation, given the presence of significantly focal lesions and prominent sarcoplasmic masses in muscle biopsies and the constant heart involvement preceding significantly the onset of the myopathy. Though a rare etiology, FLNC gene should not be excluded in early-onset arrhythmia, even in the absence of myopathy, which occurs later in the disease course.

Evaluation of serum levels and significance of soluble CD40 ligand in screening patients with hepatitis C virus-related hepatocellular carcinoma.

The study's objective was to evaluate the clinical significance of sCD40L in HCV- associated hepatocellular carcinoma (HCV-HCC) patients. Sera concentration of circulating sCD40L and IL-10 were assayed using ELISA in 30 HCVpositive patients with HCC, 30 HCV-positive patients with liver cirrhosis and 30 age-matched healthy volunteers with negative anti-HCV-Ab as a control group. Serum sCD40Lshowed statistically-significant high levels in HCV-HCC patients compared to HCV-cirrhotic patients and normal controls (P < 0.001). Serum sCD40L had higher diagnostic value in HCC patients compared with serum AFP. High sensitivity and specificity of sCD40L was observed compared to AFP (90%, 86.7% and 83% and 80% respectively). Significant positive correlation was detected between serum sCD40L and IL-10(r = 0.85 P < 0.001), AFP (r = 0.62 P < 0.05) and tumour staging (r = 0.5 P < 0.05). The study concluded that sCD40L is a valuable diagnostic tool in early diagnosis and screening for HCV and HCC as well as routine follow up of HCV cirrhosis patients. Assessment of serum IL-10 levels in HCV patients may provide a possible predictive marker for disease progression.

Mucosal delivery of antigen-coated nanoparticles to lungs confers protective immunity against tuberculosis infection in mice.

Mucosal boosting of BCG-immunised individuals with a subunit tuberculosis (TB) vaccine would be highly desirable, considering that the lungs are the principal port of entry for Mycobacterium tuberculosis (MTB) and the site of the primary infection and reactivation. However, the main roadblock for subunit TB vaccine development is the lack of suitable adjuvants that could induce robust local and systemic immune responses. Here, we describe a novel vaccine delivery system that was designed to mimic, in part, the MTB pathogen itself. The surface of yellow carnauba wax nanoparticles was coated with the highly immunogenic Ag85B Ag of MTB and they were directed to the alveolar epithelial surfaces by the incorporation of the heparin-binding hemagglutinin adhesion (HBHA) protein. Our results showed that the i.n. immunisation of BCG-primed BALB/c mice with nanoparticles adsorbed with Ag85B-HBHA (Nano-AH vaccine) induced robust humoral and cellular immune responses and IFN-γ production, and multifunctional CD4⁺ T cells expressing IFN-γ, IL-2 and TNF-α. Mice challenged with H37Rv MTB had a significantly reduced bacterial load in their lungs when compared with controls immunised with BCG alone. We therefore conclude that this immunisation approach is an effective means of boosting the BCG-induced anti-TB immunity.