Interstitial pressure and proteoglycan degradation in hydraulic- and elastase-induced lung edema.
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Animals↗
Biomedical subjects
G De Luca
Publications and source records attributed to G De Luca.
Clostridium botulinum spores and toxin in mascarpone cheese and other milk products.
A total of 1,017 mascarpone cheese samples, collected at retail, were analyzed for Clostridium botulinum spores and toxin, aerobic mesophilic spore counts, as well as pH, a(w) (water activity), and Eh (oxidation-reduction potential). In addition 260 samples from other dairy products were also analyzed for spores and botulinum toxin. Experiments were carried out on naturally and artificially contaminated mascarpone to investigate the influence of different temperature conditions on toxin production by C. botulinum. Three hundred and thirty-one samples (32.5%) of mascarpone were positive for botulinal spores, and 7 (0.8%) of the 878 samples produced at the plant involved in an outbreak of foodborne botulism also contained toxin type A. The chemical-physical parameters (pH, a(w), Eh) of all samples were compatible with C. botulinum growth and toxinogenesis. Of the other milk products, 2.7% were positive for C. botulinum spores. Growth and toxin formation occurred in naturally and experimentally contaminated mascarpone samples after 3 and 4 days of incubation at 28 degrees C, respectively.
Bacteria↗
Occurrence, removal and seasonal variation of thermophilic campylobacters and Arcobacter in sewage sludge.
The presence of thermophilic campylobacters and Arcobacter was investigated in four types of sewage sludge taken from the treatment plant in Bologna (Italy): primary, activated, thickened and anaerobically digested sludge. Campylobacter jejuni and Campylobacter coli were more numerous during the March-September period and were found only in primary sludge (22.7%) with mean counts of 278 MPN/g dry matter and 1403 MPN/g dry matter respectively. Arcobacter butzleri were found in all types of sludge with frequencies of 80% in activated and thickened sludges and 41% in digested sludges. They were more numerous in the spring/summer period with peaks in April, May, June and September. They were less sensitive to anerobic digestion than fecal bacteria, probably due to their microaerophilic growth properties. However, since they are found in anaerobically digested sludges at mean values of 7649 MPN/g dry matter the land application of digested sludges may cause high risks infection.
Campylobacter coli↗
Effect of heat and acid decontamination treatments on the recovery of Legionella pneumophila from drinking water using two selective media.
Two different decontamination systems, heat and acid, and two isolation media, GVPC and MWY agar were tested for the recovery of Legionella pneumophila from drinking water. The samples were concentrated by filtration through 0.2 micron polyamide filter and the membranes were resuspended in the original water samples. The suspension was divided into three parts: the first was placed in a 50 degrees C water bath, the second was acidified in HCl-KCl solution and the third did not undergo any treatment. The isolation was made by means of media containing charcoal, yeast extract and glycine with cycloeximide (GVPC) or vancomycin, polimixin B, anysomicin and dyes (MWY). Heating at 50 degrees C for 30 minutes was seen to be the best decontamination system above all when used with GVPC agar. Moreover, with this pretreatment higher counts were obtained both on MWY and GVPC agar. The MWY agar produced the highest isolatin percentages and the highest counts.
Acids↗
Purification and characterization of the HndA subunit of NADP-reducing hydrogenase from Desulfovibrio fructosovorans overproduced in Escherichia coli.
Based on the DNA sequence of its structural genes, clustered in the hnd operon, the NADP-reducing hydrogenase of Desulfovibrio fructosovorans is thought to be a heterotetrameric complex in which HndA and HndC constitute the NADP-reducing unit and HndD constitutes the hydrogenase unit, respectively. The weak representativity of the enzyme among cell proteins has prevented its purification. This paper discusses the purification and characterization of the HndA subunit of this unique tetrameric iron hydrogenase overproduced in Escherichia coli. The purified subunit contains 1.7 mol of non-heme iron and 1.7 mol of acid-labile sulfide/mol. EPR analysis of the reduced form of HndA indicates that it contains a single binuclear [2Fe-2S] cluster. This cluster exhibits a spectrum of rhombic symmetry with values of gx, gy, and gz equal to 1.915, 1.950, and 2. 000, respectively, and a midpoint redox potential of -395 mV. The UV-visible and EPR spectra of the [2Fe-2S] cluster indicate that HndA belongs to the [2Fe-2S] family typified by the Clostridium pasteurianum [2Fe-2S] ferredoxin. The C-terminal sequence of HndA shows 27% identity with the C-terminal sequence of the 25-kDa subunit of NADH: quinone oxidoreductase from Paracoccus denitrificans, 33% identity with the C-terminal sequence of the 24-kDa subunit from Bos taurus complex I, and 32% identity with the entire sequence of C. pasteurianum [2Fe-2S] ferredoxin. The four cysteine residues involved in HndA cluster binding have been tentatively identified on the basis of sequence identity considerations. Evidence of a HndA organization based on two independent structural domains is discussed.
Amino Acid Sequence↗
Symmetry and phase-selected NMR spectra of liquid crystalline samples.
It is demonstrated that the NMR spectra of liquid crystalline samples can be simplified by using multiple quantum filtering. In a system of N spin-12 nuclei, the N or (N-1)-multiple quantum filtered spectra (NQF or (N-1)QF) contain lines which originate only from transitions among the eigenstates belonging to the highest symmetry class of the spin permutation group. In addition the NQF spectra are divided further into two sets of lines which differ in phase by 180 degrees. A method for simulating and analysing multiple quantum filtered spectra is described, with examples from molecules with up to eight interacting spins.
Magnetic Resonance Spectroscopy↗
Effects of an anti-IL-10 monoclonal antibody on rIFNbeta-1b-mediated immune modulation. Relevance to multiple sclerosis.
The mechanism of action of recombinant IFNbeta1b (IFNbeta-1b), as a therapy for multiple sclerosis (MS), is still unknown but may result from the enhancement of ConA-induced suppressor cell function and the inhibition of IFNgamma secretion by lymphocytes. We previously demonstrated that IFNbeta-1b stimulated modest amounts of IL-10 secretion by monocytes and IL-10 activity, as cytokine synthesis inhibitory factor, was normal in MS. To determine whether IL-10 plays a role in IFNbeta-1b modulation of immune function in MS, we studied ConA-induced suppressor cell function and IFNgamma production in presence of IFNbeta-1b and an anti-IL-10 monoclonal antibody (mAb). Anti-IL-10 mAb significantly reduced the effect of IFNbeta-1b on ConA-induced suppressor cell function and IFNgamma production in healthy subjects; MS patients showed a trend of inhibition. We hypothesized that IL-10 may play a role in mediating the effects of IFNbeta-1b on suppressor cell function and IFNgamma production but suppressor molecules other than IL-10 could be also involved.
Adult↗
Smoking habits, exposure to passive smoking and attitudes to a non-smoking policy among hospital staff.
A survey was carried out into the smoking habits and exposure to passive smoking among health staff in the hospitals of Faenza, Forli and Rimini (Emilia-Romagna, Northern Italy), 2453 subjects answered anonymously a 41 question questionnaire. 53% of the subjects were professionals nurses, 16% doctors, 15% maintenance staff, 10% ancillary staff, 1% non-medical graduates, 2% were administrators and 3% were assigned to the category ¿other'. Of the subjects answering the questionnaire 39% were smokers, 19% ex-smokers and 42% non smokers. The highest number of smokers was found among women (41%) compared to men (37%) and among ancillary staff (48%) compared to nurses (41%) and doctors (31%). The males were mostly heavy smokers (> or = 20 cigarettes/d) and smoked strong cigarettes (> or = 12 mg/cig condensate content). The females were mostly light smokers (< 10 cigarettes/d) and smoked light cigarettes (1-6 mg/cig condensate content). A high percentage of subjects (87%) smoked at work especially in areas reserved for staff. 43% and 26% of shift workers and non-shift workers tended not to modify their habit when on morning or afternoon shifts. During night shifts the majority of them increased their tobacco consumption. Around 87% of hospital employees stated they were exposed to passive smoking inside the hospital especially in cooking areas, at information desks and corridors. Nurses, ancillaries and maintenance staff were those most exposed and for a greater number of hours per day compared to doctors. Almost all subjects were aware of the harm caused by passive smoking. 56% of smokers, 65% of ex-smokers and 72% of non smokers said they were willing to participate in future campaigns to limit smoking in their hospitals.
Adult↗
Proteoglycan involvement during development of lesional pulmonary edema.
We evaluated the effect of pancreatic elastase (7 IU i.v.) on pulmonary interstitial pressure (Pip) in in situ rabbit lungs by a micropuncture technique through the intact parietal pleura. Pip was -10.8 +/- 2.2 (SD) cmH2O in the control condition, increased to +5.1 +/- 1.7 cmH2O at approximately 60 min [condition referred to as mild edema (ME)], and subsequently decreased to -0.15 +/- 0.8 cmH2O, remaining steady from 80 up to 200 min with a marked increase in lung wet-to-dry weight ratio [condition referred to as severe edema (SE)], suggesting an increase in tissue compliance. We functionally correlated the measured Pip to structural modifications of proteoglycans, the major interfibrillar component of the extracellular matrix (ECM). The strength of the noncovalent bonds linking proteoglycans to other ECM components decreased with increasing severity of edema, as indicated by the increased extractability of proteoglycans with guanidine hydrochloride. Total proteoglycan recovery (expressed as microgram hexuronate/g dry tissue) increased from 436.8 +/- 14 in the control condition to 495.3 +/- 23 and 547.0 +/- 10 in ME and SE, respectively. Gel-filtration chromatography showed in ME a fragmentation of heparan sulfate proteoglycans, suggesting that elastase treatment first affected basement membrane integrity, whereas large chondroitin sulfate proteoglycans were degraded only in SE. Elastase caused a fragmentation only of the core protein of proteoglycans, the binding properties of which to collagens, fibronectin, and hyaluronic acid were markedly decreased, as indicated by a solid-phase binding assay. The sequential degradation of heparan sulfate and chondroitin sulfate proteoglycans may account for the initial increase in microvascular permeability, followed by a loss of the native architecture of the ECM, which may be responsible for the increase in tissue compliance.
Animals↗
Involvement of lung interstitial proteoglycans in development of hydraulic- and elastase-induced edema.
We extracted and isolated proteoglycans from lung tissue samples obtained from three groups of anesthetized rabbits: 1) control animals (C; n = 8) killed by overdose after 180 min; 2) animals receiving an intravenous saline infusion (S; n = 4, 1.5 ml . kg-1 . min-1) for 180 min; 3) animals receiving an intravenous bolus of 200 microg of pancreatic elastase (E; n = 4), killed after 200 min. The lung dry weight-to-wet weight ratio in the three groups was 5.2 +/- 0.2, 6.0 +/- 0.4, and 5.6 +/- 0.5, respectively. Gel-filtration analysis showed a massive fragmentation of the versican family of the extracellular matrix (ECM) in the S groups and a marked degradation of heparan sulfate-containing proteoglycans, including perlecan of the basement membrane, in the E group. The binding properties of total proteoglycans to other ECM components were lowered in both groups relative to control. The decrease in proteoglycan binding was more pronounced for collagen type IV in the E group relative to C (-93.5%, P < 0.05) and for hyaluronic acid in the S groups (-85.8%, P < 0.05). These findings suggest that elastase treatment produces a major degree of damage to the organization of basement membrane, whereas saline loading affects more markedly the architecture of interstitial ECM. Qualitative zymography performed on lung extracts showed increased gelatinase activities in both S and E groups, providing direct evidence that the activation of tissue proteinases may play a role in acute lung injury.
Animals↗
Occurrence of Listeria monocytogenes in sewage sludge.
The search for salmonella is the only bacteriological index included in Italian guidelines (in accordance with EU regulations) for the use of sewage sludge in agriculture. As a result, information regarding the presence of Listeria monocytogenes is rather limited. We therefore decided to carry out an investigation of Listeria in the sludge produced by the Bologna (Italy) treatment plant during the various phases of treatment. Five different types of sludge were analysed (primary raw, activated, thickened, digested and dewatered) in a total of 66 samples. The highest frequency and concentrations of Listeria species (100% and 2,743 MPN/g dry matter) and the lowest (63% and 6 MPN/g dry matter) were found in the activated and digested sludge respectively. These bacteria were mostly present in spring and autumn and positively correlated only with fecal streptococci. Four species were isolated: Listeria monocytogenes, Listeria innocua, Listeria welshimeri and Listeria grayi. Listeria monocytogenes (prevalent serotype 4b) was seen to be resistent to the biological oxidation but sensitive to anaerobic conditions during thickening and digestion. The dewatering process led to an increase in contamination. Since the sludge is used to fertilize land destined for vegetable farming our results show that it may represent a potential health risk.
Enterobacteriaceae↗
Identification of viable myocardium in patients with chronic coronary artery disease using rest-redistribution thallium-201 tomography: optimal image analysis.
UNLABELLED: With the widely used 50% threshold, sensitivity is high, but specificity is low in detecting viable myocardium on 201Tl SPECT. In this study, we sought to identify the best threshold for semiquantitative 201Tl analysis. METHODS: Rest-redistribution 201Tl SPECT was performed in 46 patients with chronic coronary artery disease before and after myocardial revascularization. Regional function was evaluated by two-dimensional echocardiography before and after myocardial revascularization using a 3-point scale (1 = normal, 2 = hypokinetic, 3 = a/dyskinetic). Myocardial segments with abnormal systolic function were defined as viable if the systolic function score decreased > or = 1 after myocardial revascularization. A second group of 12 patients with chronic coronary artery disease constituted the validation population. Sensitivity-specificity curves, as well as receiver operating characteristic curves, for rest and redistribution mages were generated by varying the 201Tl uptake threshold. RESULTS: A 65% threshold uptake using resting images was found to be the best for detecting a/dyskinetic segments that improve after myocardial revascularization from those that do not improve. Sensitivity was lower with a 65% threshold (75%) than with a 50% threshold (90%, p < 0.05), but specificity was higher (76% versus 26%, p < 0.05) resulting in better accuracy (76% versus 57%, p < 0.05) and positive predictive value (77% versus 55%), while the negative predictive value was not different (69% versus 75%, p not significant). The area under the receiver operating characteristic curve was significantly (p < 005) larger for rest (0.80 +/- 0.05) as opposed to redistribution (0.72 +/- 0.05) images. Similar results were obtained in a subgroup of patients with low ejection fraction. Significant correlations between the percentage of revascularized viable segments and both the change in ejection fraction and in postrevascularization ejection fraction were found. When these findings were applied in the validation group, a gain in specificity, accuracy and positive predictive value was obtained with the 65% threshold compared with the 50% threshold. CONCLUSION: This study demonstrated that analysis of resting images and use of the 65% 201Tl uptake threshold is preferable for separating viable from not viable dyssynergic myocardial segments in patients with chronic coronary artery disease.
Case-Control Studies↗
Modifications of proteoglycans extracted from monolayer cultures of young and senescent human skin fibroblasts.
Proteoglycans (PGs) were extracted from culture monolayers of human skin fibroblasts (HFs) at early and late passages. Total PGs from senescent cells had markedly reduced abilities to bind type I collagen and hyaluronic acid, but retained normal binding properties with fibronectin and laminin. The constituent polysaccharides of PGs were comparatively characterised. PGs recovered from young and senescent HF cultures had equivalent total polyanionic charges and similar size distributions of the glycosaminoglycan chains. This applied to both types of polysaccharide chains found in PGs, namely the galactosaminoglycuronans (GalN-GAGs) and the glucosaminoglycuronans (GlcN-GAGs). However, senescent HFs produced a greater proportion of PGs containing GlcN-GAG chains and increased the sulphation of the remaining PG fraction with GalN-GAG moieties, yielding a major gain of C6-sulphate groups in the galactosamine residues.
Aging↗
Staphylococcus aureus in dairy products in the Bologna area.
A study was carried out to evaluate the extent of contamination by Staphylococcus spp. and in particular by Staphylococcus aureus in dairy products on sale in the Bologna area. Staphylococcus aureus was found in 16.30% of the 135 cheese samples examined: 8.3% in soft cheese, 17.60% in blue cheese, 18.9% in semi-soft cheese and 25.0% in mozzarella-type cheese. The mean concentration was 2699 CFU/g in the 22 positive samples. The most commonly found species were Staphylococcus epidermidis (14.8%), Staphylococcus hominis (19.5%), Staphylococcus xylosus 2 (19.2%) and Staphylococcus cohnii (16.3%). Staphylococcus aureus was found mostly during the hot months while the other common species were found mostly in the period October-March.
Animals↗
Chondroitin 4-sulphate exhibits inhibitory effect during Cu2+-mediated LDL oxidation.
Chondroitin 4-sulphate (C4S), a basic component of the extracellular matrix of the artery wall, inhibited copper-induced low density lipoprotein (LDL) oxidation by prolonging the lag time and reducing the rate of propagation. LDL oxidation was assessed by monitoring conjugated dienes and low level chemiluminescence. A possible initial key reaction in LDL oxidation, the reduction of copper(II) to copper(I) by LDL, was decreased in the presence of C4S. Moreover, C4S protected tryptophan residues of Apo-B-100 in the early stage of LDL oxidation and during the subsequent propagation phase. The presence of the sulphate group in position 4 of N-acetylgalactosaminyl residues of C4S is crucial for protective activity. In fact, the structurally related chondroitin 6-sulphate, also present in tissues, had no effect on LDL oxidation. These data suggest that C4S may contribute to protect LDL against oxidation in arterial intima.
Adult↗
Modifications of proteoglycans secreted into the growth medium by young and senescent human skin fibroblasts.
The properties of proteoglycans (PGs) secreted into the growth medium by normal young and senescent human skin fibroblasts (HFs) were investigated. In both cases, the incorporation per cell of radioactive precursors into total PGs was similar. The polysaccharide chains of PGs from young and senescent HFs were mainly represented by galactosaminoglycuronans and showed a similar range of size distribution. However, galactosaminoglycuronans of PGs secreted by senescent HFs had a lower content of unsulphated disaccharides and a lower proportion of D-glucuronosyl residues. Moreover, senescent HFs released into the growth medium higher relative amounts of small PGs with chondroitin sulphate, dermatan sulphate chains, such as decorin.
Cell Division↗
Inflammatory myopathy after intravenous streptokinase.
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Aged↗
Physiological characteristics and growth behavior of single and double hydrogenase mutants of Desulfovibrio fructosovorans.
The presence of one periplasmic [NiFe] hydrogenase, one periplasmic [Fe] hydrogenase, and one cytoplasmic NADP-reducing hydrogenase has been previously established in Desulfovibrio fructosovorans. In the present work, marker-exchange mutagenesis was performed to determine the function of the tetrameric NADP-reducing hydrogenase encoded by the hndA, B, C, and D genes. The mutations performed were not lethal to the cells, although the H2-dependent NADP reduction was completely abolished. The double-mutated DM4 (DeltahynABC, DeltahndD) strain was still able to grow on hydrogen plus sulfate as the sole energy source. The growth may have occurred under these culture conditions because of the presence of the remaining [Fe] hydrogenase. The cells grew differently on various substrates depending on whether fructose, lactate, or pyruvate was used in the presence of sulfate. The (hnd mutant growth rates were 25-70% lower than those of the wild-type strain, although the molar growth yield remained unchanged. By contrast, mutants devoid of both [NiFe] hydrogenase and NADP-reducing hydrogenase had 24-38% lower growth yields and showed a corresponding drop in the growth rates. We concluded that each of the three hydrogenases may contribute to the energy supply in D. fructosovorans and that the loss of one enzyme might be compensated for by another. However, the loss of two hydrogenases affected the phosphorylation accompanying the metabolism of fructose, lactate, and pyruvate.
Adenosine Triphosphate↗