Pseudomonas aeruginosa bloodstream infections: risk factors and treatment outcome related to expression of the PER-1 extended-spectrum beta-lactamase

BACKGROUND: Bloodstream infection (BSI) due to Pseudomonas aeruginosa (Pa) has relevant clinical impact especially in relation to drug resistance determinants. The PER-1 extended-spectrum beta-lactamase (ESBL) is a common enzyme conferring high-level resistance to anti-pseudomonal cephalosporins. Risk factors and treatment outcome of BSI episodes caused by PER-1-positive Pa (PER-1-Pa) strains were compared to those caused by ESBL-negative Pa isolates (ESBL-N-Pa). METHODS: Twenty-six BSI cases due to ceftazidime-resistant Pa strains have been investigated. MIC values of anti-pseudomonal drugs were determined by the Etest method (AB Biodisk, Solna, Sweden). The double-disk synergy test was used to detect ESBL production. PCR amplification and DNA sequencing were used to characterize ESBL types. Clinical records of BSI-patients were examined retrospectively. Demographic data, underlying diseases (McCabe-Jackson classification and Charlson weighted index), risk factors, antimicrobial therapy, and treatment outcome were evaluated in cases due to ESBL-positive and cases due to ESBL-N-Pa isolates. Unpaired Student's t-test, Mann-Whitney U-test, Fisher's exact test and the χ(2 )test were used for statistical analysis. RESULTS: Nine Pa isolates expressed the PER-1 ESBL; the remaining 17 isolates did not produce ESBLs. Severe sepsis (P = 0.03), bladder and intravascular catheters (both P = 0.01), immunosuppressive therapy (P = 0.04), and mechanical ventilation (P = 0.03) were significantly associated with BSI due to PER-1-Pa. Empirical treatment (P = 0.02) and treatment after ID/AST (P < 0.01) were rarely adequate in PER-1-Pa cases. With regard to treatment outcome, 77.8% BSI cases due to PER-1-Pa vs. 28.6% cases due to ESBL-N-Pa isolates failed to respond (P < 0.03). All cases due to PER-1-Pa that were treated with carbapenems (alone or in combination with amikacin) failed to respond. In contrast, 7/8 cases due to ESBL-N-Pa given carbapenems were responders. CONCLUSION: Therapeutic failure and increased hospital costs are associated with BSI episodes caused by PER-1-Pa strains. Thus, recognition and prompt reporting of ESBL-production appears a critical factor for the management of patients with serious P. aeruginosa infections

Culture of skeletal myoblasts from human donors aged over 40 years: dynamics of cell growth and expression of differentiation markers

BACKGROUND: Local myogenesis, neoangiogenesis and homing of progenitor cells from the bone marrow appear to contribute to repair of the infarcted myocardium. Implantation into heart tissues of autologous skeletal myoblasts has been associated with improved contractile function in animal models and in humans with acute myocardial ischemia. Since heart infarction is most prevalent in individuals of over 40 years of age, we tested whether culture methods available in our laboratory were adequate to obtain sufficient numbers of differentiated skeletal myoblasts from muscle biopsy specimens obtained from patients aged 41 to 91. METHODS AND RESULTS: No matter of donor age, differentiated skeletal muscle cells could be produced in vitro in amounts adequate for cellular therapy (≥300 millions). Using desmin as a cytoplasmic marker, about 50% cultured cells were differentiated along myogenic lineages and expressed proteins proper of skeletal muscle (myosin type I and II, actin, actinin, spectrin and dystrophin). Cytogenetic alterations were not detected in cultured muscle cells that had undergone at least 10 population doublings. Molecular methods employed for the screening of persistent viral infections evidenced that HCV failed to replicate in muscle cells cultured from one patient with chronic HCV infection. CONCLUSION: The proposed culture methods appear to hold promise for aged patients not only in the field of cardiovascular medicine, but also in the urologic and orthopedic fields

Sulfate Reducing Bacteria as Bio-cleaning Agents: Development of New Methodologies and Study Cases

In the last decades, the contribution of different scientific disciplines in the field of restoration and conservation of cultural heritage to finding alternative methods of investigation that are even more effective and fully respect artworks, operators and environment, has greatly increased. An example is the University of Milan patent that provides for the use of some specialized bacteria as cleaning agents. This method, known as biocleaning, is based on the use of Desulfovibrio vulgaris, a sulfate reducing bacterium, for the removal of sulfate alterations from stone surfaces. Recently, the university spin-off Micro4yoU purchased the patent by initiating a series of investments aimed at enhancing the commercial product from prototype. The present work describes: the desulfation mechanism operated by Desulfovibrio vulgaris and the technological shift necessary to obtain a biological formulation usable in situ, with two practical case studies

Sulfate Reducing Bacteria as Bio-cleaning Agents: Development of New Methodologies and Study Cases

In the last decades, the contribution of different scientific disciplines in the field of restoration and conservation of cultural heritage to finding alternative methods of investigation that are even more effective and fully respect artworks, operators and environment, has greatly increased.An example is the University of Milan patent that provides for the use of some specialized bacteria as cleaning agents. This method, known as biocleaning, is based on the use of Desulfovibrio vulgaris, a sulfate reducing bacterium, for the removal of sulfate alterations from stone surfaces.Recently, the university spin-off Micro4yoU purchased the patent by initiating a series of investments aimed at enhancing the commercial product from prototype. The present work describes: the desulfation mechanism operated by Desulfovibrio vulgaris and the technological shift necessary to obtain a biological formulation usable in situ, with two practical case studies.Negli ultimi decenni è cresciuto il contributo che le diverse discipline scientifiche hanno dato al recupero e alla conservazione del patrimonio artistico per trovare soluzioni sempre più efficaci e rispettose del bene, dell’operatore e dell’ambiente.Un esempio è un brevetto sviluppato dall’Università degli studi di Milano che prevede l’utilizzo di alcuni specifici batteri come agenti di pulitura. Tale metodo, indicato col termine di bio-pulitura propone, tra gli altri, l’impiego del batterio solfato riduttore Desulfovibrio vulgaris per la rimozione di alterazioni solfatiche da superfici litoidi. Recentemente, il brevetto è stato acquisito dallo spin-off universitario Micro4yoU che ha avviato una serie di investimenti per sviluppare, a partire dal prototipo di laboratorio, un prodotto commerciale.Nel presente lavoro verranno illustrati: il meccanismo di desolfatazione ad opera di Desulfovibrio vulgaris, il processo di trasferimento tecnologico affrontato per ottenere un formulato microbico utilizzabile in cantiere e due casi studio applicativi

Effects of ω-3 PUFA-enriched diet on growth parameters in a syngenic murine model of breast adenocarcinoma: a possible role for estrogen receptor alpha

Background: Breast cancer (BC) is the most common tumour among women and 75% of BC are estrogen receptor (ER)dependent. In particular, ERα promotes tumour growth, while ERβ has an anti-proliferative effect [1]. Epidemiological data have linked ω-3 polyunsaturated fatty acid (PUFA) consumption to lower incidence of BC and several experimental studies showed the anti-proliferative effects of ω-3 fish oil in different tumour models [2,3]. Chia seed oil is rich in α-linolenic acid (ALA 18:3 ω-3), while corn oil is rich in linoleic acid (LA 18:2 ω-6), precursors of eicosapentaenoic acid (EPA) and arachidonic acid (AA), respectively. Based on substrate availability, these FAs give rise to different eicosanoid signatures with opposite effects in cancer [4]. ω-3 PUFAs generate both anti-inflammatory prostanoids and reactive oxygen species (ROS), which in turn could affect NF-κB. Indeed, NF-κB belongs to a family of transcription factors with a key role in inflammation and oxidative stress, but its role in tumour development is still controversial [5]. The aim of the study was to determine possible processes that are activated by dietary lipids regulating BC growth and metastasis.Fil: Vara Messler, Marianela. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas. Instituto de Biología Celular; Argentina.Fil: Vara Messler, Marianela. Universidad de Padova. Departamento de Ciencias Farmacéuticas y Farmacológicas; Italia.Fil: Pasqualini, María Eugenia. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas. Instituto de Biología Celular; Argentina.Fil: Comba, Andrea. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas. Instituto de Biología Celular. Cátedra de Biología, Histología y Embriología; Argentina.Fil:Comba, Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ciencias de la Salud; Argentina.Fil: Toniolo, Alicia. Universidad de Padova. Departamento de Ciencias Farmacéuticas y Farmacológicas; Italia.Fil: Trenti, Annalisa. Universidad de Padova. Departamento de Ciencias Farmacéuticas y Farmacológicas; Italia.Fil: Quiroga, Patricia. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas. Instituto de Biología Celular; Argentina.Fil: Valentich, Mirta Ana. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas. Instituto de Biología Celular; Argentina.Fil: Valentich, Mirta Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ciencias de la Salud; Argentina.Fil: Bolego, Chiara. Universidad de Padova. Departamento de Ciencias Farmacéuticas y Farmacológicas; Italia.Otras Ciencias de la Salu

A portable setup for the voltammetric determination of total mercury in fish with solid and nanostructured gold electrodes

A simple procedure for field fish sample pretreatment was developed. This treatment in combination with square wave anodic stripping voltammetry (SW-ASV) with solid gold electrodes (SGE) and gold nanoparticle-modified glassy carbon electrodes (AuNPs-GCE) was applied for the determination of total mercury content. A certified reference material (CRM, Tuna Fish BCR 463), ten freeze-dried samples of canned tuna and two fresh fish samples were analysed both with a bench-top voltammetric analyser after microwave digestion and with a portable potentiostat after mild eating using a small commercial food warmer. The results obtained by the two SW-ASV approaches and by a Direct Mercury Analyser (DMA), the official method for mercury determination, were in very good agreement. In particular, (i) the results obtained with in field procedure are consistent with those obtained with the conventional microwave digestion; (ii) the presence of gold nanoparticles on the active electrode surface permits an improvement of the analytical performance in comparison to the SGE: the Limit of Quantification (LOQ) for mercury in fish-matrix was 0.1 &#956;g L&#8722;1 (Hg cell concentration), corresponding to 0.06 mg kg&#8722;1 wet fish, which is a performance comparable to that of DMA. The pretreatment proposed in this study is very easy and applicable to fresh fish; in combination with a portable potentiostat, it proved to be an interesting procedure for on-site mercury determination

Characterization of extracellular vesicles in osteoporotic patients compared to osteopenic and healthy controls

Extracellular vesicles (EVs) are mediators of a range of pathological conditions. However, their role in bone loss disease has not been well understood. In this study we characterized plasma EVs of 54 osteoporotic (OP) postmenopausal women compared to 48 osteopenic (OPN) and 44 healthy controls (CN), and we investigated their effects on osteoclasts and osteoblasts. We found no differences between the three groups in terms of anthropometric measurements and biochemical evaluation of serum calcium, phosphate, creatinine, PTH, 25-hydroxy vitamin D and bone biomarkers, except for an increase of CTX level in OP group. FACS analysis revealed that OP patients presented a significantly increased number of EVs and RANKL(+) EVs compared with both CN and OPN subjects. Total EVs are negatively associated with the lumbar spine T-score and femoral neck T-score. Only in the OPN patients we observed a positive association between the total number of EVs and RANKL(+) EVs with the serum RANKL. In vitro studies revealed that OP EVs supported osteoclastogenesis of healthy donor peripheral blood mononuclear cells at the same level observed following RANKL and M-CSF treatment, reduced the ability of mesenchymal stem cells to differentiate into osteoblasts, while inducing an increase of OSTERIX and RANKL expression in mature osteoblasts. The analysis of miRNome revealed that miR-1246 and miR-1224-5p were the most upregulated and downregulated in OP EVs; the modulated EV-miRNAs in OP and OPN compared to CN are related to osteoclast differentiation, interleukin-13 production and regulation of canonical WNT pathway. A proteomic comparison between OPN and CN EVs evidenced a decrease in fibrinogen, vitronectin, and clusterin and an increase in coagulation factors and apolipoprotein, which was also upregulated in OP EVs. Interestingly, an increase in RANKL(+) EVs and exosomal miR-1246 was also observed in samples from patients affected by Gorham-Stout disease, suggesting that EVs could be good candidate as bone loss disease biomarkers. (c) 2022 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR)

An electrochemical gas sensor based on paper supported room temperature ionic liquids

A sensitive and fast-responding membrane-free amperometric gas sensor is described, consisting of a small filter paper foil soaked with a room temperature ionic liquid (RTIL), upon which three electrodes are screen printed with carbon ink, using a suitable mask. It takes advantage of the high electrical conductivity and negligible vapour pressure of RTILs as well as their easy immobilization into a porous and inexpensive supporting material such as paper. Moreover, thanks to a careful control of the preparation procedure, a very close contact between the RTIL and electrode material can be achieved so as to allow gaseous analytes to undergo charge transfer just as soon as they reach the three-phase sites where the electrode material, paper supported RTIL and gas phase meet. Thus, the adverse effect on recorded currents of slow steps such as analyte diffusion and dissolution in a solvent is avoided. To evaluate the performance of this device, it was used as a wall-jet amperometric detector for flow injection analysis of 1-butanethiol vapours, adopted as the model gaseous analyte, present in headspace samples in equilibrium with aqueous solutions at controlled concentrations. With this purpose, the RTIL soaked paper electrochemical detector (RTIL-PED) was assembled by using 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl) imide as the wicking RTIL and printing the working electrode with carbon ink doped with cobalt(II) phthalocyanine, to profit from its ability to electrocatalyze thiol oxidation. The results obtained were quite satisfactory (detection limit: 0.5 mu M; dynamic range: 2-200 mu M, both referring to solution concentrations; correlation coefficient: 0.998; repeatability: +/- 7% RSD; long-term stability: 9%), thus suggesting the possible use of this device for manifold applications.Italian Ministry of University and Scientific ResearchItalian Ministry of University and Scientific Researc

Phytochemical analysis and In vitro antileukemic activity of alkaloid-enriched extracts from Vinca sardoa (Stearn) Pignatti

Vinca sardoa (Stearn) Pignatti, known as Sardinian periwinkle, is widely diffused in Sardinia (Italy). This species contains indole alkaloids, which are known to have a great variety of biological activities. This study investigated the antileukemic activity against a B lymphoblast cell line (SUP-B15) of V. sardoa alkaloid-rich extracts obtained from plants grown in Italy, in Iglesias (Sardinia) and Rome (Latium). All the extracts showed a good capacity to induce reductions in cell proliferation of up to 50% at the tested concentrations (1–15 g/mL). Moreover, none of the extracts showed cytotoxicity on normal cells at all the studied concentrations

APPLICAZIONI DELLA METODOLOGIA LCA NEL CAMPO DELLA GESTIONE E DEL TRATTAMENTO DEI RIFIUTI

L\u2019articolo nasce da un\u2019iniziativa del gruppo di lavoro \u201cGestione e trattamento dei rifiuti\u201d dell\u2019Associazione Rete Italiana LCA (Life Cycle Assessment). L\u2019obiettivo principale del gruppo di lavoro \ue8 quello di ampliare le conoscenze relative alla specificit\ue0 della metodologia LCA applicata ai processi di gestione e di trattamento dei rifiuti. L\u2019articolo riporta otto casi studio condotti da alcuni dei partecipanti al gruppo di lavoro, con il duplice obiettivo di mostrare il vasto campo di applicazione della metodologia LCA pur mantenendosi all\u2019interno del settore rifiuti e le specificit\ue0 dell\u2019applicazione di tale metodologia ai processi di gestione e trattamento dei rifiuti. Gli otto casi studio sono stati raggruppati per macro-finalit\ue0 dell\u2019applicazione, ossia: LCA applicata per valutare i benefici ottenibili dal recupero di varie tipologie di rifiuti, LCA a supporto della pianificazione regionale e/o nazionale e LCA a supporto dell\u2019economia circolare