Genotypic and phenotypic heterogeneity in Alicyclobacillus acidoterrestris: a contribution to species characterization

Alicyclobacillus acidoterrestris is the main cause of most spoilage problems in fruit juices and acidic products. Since soil borne species often contaminate fruit juices and do not need strict extreme requirements for survival, it is a great concern to investigate whether and how soil species could evolve from their ecological niches in microbial community to new environments as fruit juices. In this study, 23 isolates of thermo-acidophilic, spore-forming bacteria from soil were characterized by cultural and molecular methods. In addition, 2 strains isolated from a spoilage incident in pear juice were typed. Strains phenotyping showed that they could be grouped into 3 different clusters, and some isolates showed identical or quite similar patterns. Analyzing pH and temperature ranges for growth, the majority of strains were able to grow at values described for many species of Alicyclobacillus. Qualitative utilization of lysine, arginine and indole production from tryptophan revealed, for the first time, deamination of lysine and decarboxylation of arginine. Resistance to 5% NaCl as well as the ability to hydrolyze starch and gelatin, nitrate reduction, catalase and oxidase activities confirmed literature evidences. Examining of 16S rRNA, showed that isolates were divided into three blocks represented by effectively soil species and strains that are moving from soil to other possible growing source characterized by parameters that could strongly influence bacterial survival. RAPD PCR technique evidenced a great variability in banding patterns and, although it was not possible to obtain genotypically well-distinguished groups, it was feasible to appreciate genetic similarity between some strains. In conclusion, the investigation of a microbial community entails a combination of metagenomic and classic culture-dependent approaches to expand our knowledge about Alicyclobacillus and to look for new subspecies

Predictors of cardiovascular disease in asthma and chronic obstructive pulmonary disease

BACKGROUND: Cardiovascular disease (CVD) is a common comorbidity in patients with chronic airway obstruction, and is associated with systemic inflammation and airway obstruction. The aim of this study was to evaluate the predictors of CVD in two different conditions causing chronic airway obstruction, asthma and COPD. METHODS: Lung function tests, clinical and echocardiographic data were assessed in 229 consecutive patients, 100 with asthma and 129 with COPD. CVD was classified into: pressure overload (PO) and volume overload (VO). Sub-analysis of patients with ischemic heart disease (IHD) and pulmonary hypertension (PH) was also performed. RESULTS: CVD was found in 185 patients (81%: 51% COPD and 30% asthmatics) and consisted of PO in 42% and of VO in 38% patients. COPD patients, as compared to asthmatics, had older age, more severe airway obstruction, higher prevalence of males, of smokers, and of CVD (91% vs 68%), either PO (46% vs 38%) or VO (45% vs 30%). CVD was associated with older age and more severe airway obstruction both in asthma and COPD. In the overall patients the predictive factors of CVD were age, COPD, and male sex; those of PO were COPD, BMI, VC, FEV(1) and MEF(50) and those of VO were age, VC and MEF(50). In asthma, the predictors of CVD were VC, FEV(1), FEV(1) /VC%, and PaO(2), those of PO were VC, FEV(1) and FEV(1) /VC%, while for VO there was no predictor. In COPD the predictors of CVD were age, GOLD class and sex, those of VO age, VC and MEF(50), and that of PO was BMI. Sub-analysis showed that IHD was predicted by COPD, age, BMI and FEV(1), while PH (found only in 25 COPD patients), was predicted by VO (present in 80% of the patients) and FEV(1). In subjects aged 65 years or more the prevalence of CVD, PO and VO was similar in asthmatic and COPD patients, but COPD patients had higher prevalence of males, smokers, IHD, PH, lower FEV(1) and higher CRP. CONCLUSIONS: The results of this study indicate that cardiovascular diseases are frequent in patients with chronic obstructive disorders, particularly in COPD patients. The strongest predictors of CVD are age and airway obstruction. COPD patients have higher prevalence of ischemic heart disease and pulmonary hypertension. In the elderly the prevalence of PO and VO in asthma and COPD patients is similar

Impact of gluten-friendly bread on the metabolism and function of in vitro gut microbiota in healthy human and coeliac subjects

The main aim of this paper was to assess the in vitro response of healthy and coeliac human faecal microbiota to gluten-friendly bread (GFB). Thus, GFB and control bread (CB) were fermented with faecal microbiota in pH-controlled batch cultures. The effects on the major groups of microbiota were monitored over 48 h incubations by fluorescence in situ hybridisation. Short-chain fatty acids (SCFAs) were measured by high-performance liquid chromatography (HPLC). Furthermore, the death kinetics of Lactobacillus acidophilus, Bifidobacterium animalis subsp. lactis, Staphylococcus aureus, and Salmonella Typhimurium in a saline solution supplemented with GFB or CB were also assessed. The experiments in saline solution pinpointed that GFB prolonged the survival of L. acidophilus and exerted an antibacterial effect towards S. aureus and S. Typhimurium. Moreover, GFB modulated the intestinal microbiota in vitro, promoting changes in lactobacilli and bifidobacteria members in coeliac subjects. A final multivariate approach combining both viable counts and metabolites suggested that GFB could beneficially modulate the coeliac gut microbiome; however, human studies are needed to prove its efficacy

Therapeutic potential of MEK inhibition in acute myelogenous leukemia: rationale for "vertical" and "lateral" combination strategies

: In hematological malignancies, constitutive activation of the RAF/MEK/ERK pathway is frequently observed, conveys a poor prognosis, and constitutes a promising target for therapeutic intervention. Here, we investigated the molecular and functional effects of pharmacological MEK inhibition in cell line models of acute myeloid leukemia (AML) and freshly isolated primary AML samples. The small-molecule, ATP-non-competitive, MEK inhibitor PD0325901 markedly inhibited ERK phosphorylation and growth of several AML cell lines and approximately 70 % of primary AML samples. Growth inhibition was due to G(1)-phase arrest and induction of apoptosis. Transformation by constitutively active upstream pathway elements (HRAS, RAF-1, and MEK) rendered FDC-P1 cells exquisitely prone to PD0325901-induced apoptosis. Gene and protein expression profiling revealed a selective effect of PD0325901 on ERK phosphorylation and compensatory upregulation of the RAF/MEK and AKT/p70( S6K ) kinase modules, potentially mediating resistance to drug-induced growth inhibition. Consequently, in appropriate cellular contexts, both "vertical" (i.e., inhibition of RAF and MEK along the MAPK pathway) and "lateral" (i.e., simultaneous inhibition of the MEK/ERK and mTOR pathways) combination strategies may result in synergistic anti-leukemic effects. Overall, MEK inhibition exerts potent growth inhibitory and proapoptotic activity in preclinical models of AML, particularly in combination with other pathway inhibitors. Deeper understanding of the molecular mechanisms of action of MEK inhibitors will likely translate into more effective targeted strategies for the treatment of AML

Dynamic accommodation without feedback does not respond to isolated blur cues.

The aim of this study was to determine whether dynamic accommodation responds to isolated blur cues without feedback, and without changes in the distance of the object. Nine healthy subjects aged 21-40years were recruited. Four different aberration patterns were used as stimuli to induce blur with (1) the eye's natural, uncorrected, optical aberrations, (2) all aberrations corrected, (3) spherical aberration only, or (4) astigmatism only. The stimulus was a video animation based on computer-generated images of a monochromatic Maltese cross. Each individual video was generated for each subject off-line, after measuring individual aberrations at different accommodation levels. The video simulated sinusoidal changes in defocus at 0.2Hz. Dynamic images were observed through a 0.8mm pinhole placed at a plane conjugated with the eye's pupil, thus effectively removing potential feedback stemming from accommodation changes. Accommodation responses were measured with a Hartmann-Shack aberrometer for the four different aberration patterns. The results showed that seven out of nine subjects did not respond to any stimuli, whereas the response of the other two subjects was erratic and they seemed to be searching rather than following the stimulus. A significant reduction in average accommodative gain (from 0.52 to 0.11) was obtained when the dioptric demand cue was removed. No statistically significant differences were found among the experimental conditions used. We conclude that aberration related blur does not drive the accommodation response in the absence of feedback from accommodation

...A tre giorni di cammino da Napoli. L'ascesa di una famiglia patrizia di Capitanata : i Tontoli di Manfredonia tra XVI e XVIII secolo

Antonio Ciuffreda, « ... A tre giorni di cammino da Napoli». L'ascesa di una famiglia patrizia di Capitanata : i Tontoli di Manfredonia tra XVI e XVIII secolo, p. 165-216. L'indagine condotta pone al centro dell'analisi le scelte compiute e la rete di alleanze allacciate da una famiglia del patriziato cittadino meridionale tra XVI e XVIII secolo. Alla base della politica familiare vi è una forte e duratura solidarietà interna unita ad una attenta strategia matrimoniale e clientelare. Dall'affitto e gestione di masserie di campo si passa, nel corso degli anni, al controllo dello spazio cittadino di intermediazione commerciale e al sopraggiungere della crisi, si privilegia il rapporto con il mondo ecclesiastico che consente di allargare la sfera di relazioni sociali e familiari. Assunto uno 'status' che si ispira ad atteggiamenti propri della nobiltà titolata, dal microcosmo cittadino provinciale la (v. retro) famiglia si sposta verso la capitale, percorrendo quei «tre giorni di cammino» che li separa da Napoli.Ciuffreda Antonio...A tre giorni di cammino da Napoli. L'ascesa di una famiglia patrizia di Capitanata : i Tontoli di Manfredonia tra XVI e XVIII secolo. In: Mélanges de l'École française de Rome. Italie et Méditerranée, tome 103, n°1. 1991. pp. 165-216