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Biomedical subjects

G Spano

Publications and source records attributed to G Spano.

At least 19 recordsLinked to original sources
PubMed2006

A comparative study on the value of FDG-PET and sentinel node biopsy to identify occult axillary metastases.

BACKGROUND: Sentinel node biopsy (SNB) has become a standard treatment in staging axillary lymph nodes in early breast cancer. SNB, however, is an invasive procedure and is time-consuming when the sentinel node is analysed intra-operatively. Breast cancer is frequently characterised by increased 2-fluoro-2-deoxy-D-glucose uptake and many studies have shown encouraging results in detecting axillary lymph node metastases. The aim of this study was to compare SNB and -positron emission tomography (-PET) imaging, to assess their values in detecting occult axillary metastases. PATIENTS AND METHODS: In all, 236 patients with breast cancer and clinically negative axilla were enrolled in the study. 18-FDG-PET was carried out before surgery, using a positron emission tomography (PET)/computed tomography scanner. In all patients, SNB was carried out after identification through lymphoscintigraphy. Patients underwent axillary lymph nodes dissection (ALND) in cases of positive FDG-PET or positive SNB. The results of PET scan were compared with histopathology of SNB and ALND. RESULTS: In all, 103 out of the 236 patients (44%) had metastases in axillary nodes. Sensitivity of FDG-PET scan for detection of axillary lymph node metastases in this series was low (37%); however, specificity and positive predictive values were acceptable (96% and 88%, respectively). CONCLUSIONS: The high specificity of PET imaging indicates that patients who have a PET-positive axilla should have an ALND rather than an SNB for axillary staging. In contrast, FDG-PET showed poor sensitivity in the detection of axillary metastases, confirming the need for SNB in cases where PET is negative in the axilla.

Adult↗
PubMed2006

In Vivo PCR-DGGE analysis of Lactobacillus plantarum and Oenococcus oeni populations in red wine.

In order to monitor Lactobacillus plantarum and Oenococcus oeni in red wine produced with Italian grape (variety "Primitivo di Puglia"), a polymerase chain reaction- denaturing gradient gel electrophoresis (PCR-DGGE) approach using the rpoB as gene target was established. Wine was treated or not with potassium metabisulphite and supplemented with a commercial bacterial starter of O. oeni to encourage malolactic fermentation. Samples were taken from the vinification tanks at 4, 10, 16, 22, and 28 days after the start of alcoholic fermentation. Genomic DNA was directly isolated from wine and identification of lactic acid bacteria was performed using primers rpoB1, rpoB1O, and rpoB2 able to amplify a region of 336 bp corresponding to the rpoB gene. Amplified fragments were separated in a 30-60% DGGE gradient, and the ability of the PCR-DGGE analysis to distinguish L. plantarum and O. oeni was assessed. The results reported suggest that the PCR-DGGE method, based on the rpoB gene as molecular marker, is a reproducible and suitable tool and may be of great value for wine makers in order to monitor spoilage microorganisms during wine fermentation.

DNA, Bacterial↗
PubMed2006

Environmental stress response in wine lactic acid bacteria: beyond Bacillus subtilis.

Lactic acid bacteria (LAB) constitute a heterogeneous group of bacteria that are traditionally used to produce fermented foods. The industrialization of food transformations has increased the economical importance of LAB, as they play a crucial role in the development of the organoleptic and hygienic quality of fermented products. However, the strains selected for industrial purposes, should tolerate adverse conditions encountered in industrial processes, either during starter handling and storage (freeze-drying, freezing, or spray-drying) or during food processing in which abiotic stresses such as heat, cold, acidity, and high concentration of NaCl or ethanol are common. Wine LAB have to deal with several stresses including an acidic pH, a high alcoholic content, non optimal growth temperatures, and growth-inhibitory compounds such as fatty acids and tannins, originated from yeast and bacteria metabolism. Wine LAB have developed several mechanisms to escape or to tolerate wine conditions. They carry out a malolactic fermentation in this stressful environment. In addition to the regulation of the expression of specific genes, bacteria have evolved adaptive networks to face the challenges of a changing environment and to survive under conditions of stress. The so called Global Regulatory Systems control the simultaneous expression of a large number of genes in response to a variety of environmental stress factors. CIRCE sequences able to bind the HrcA repressor, sigma(B) dependent promoters and CtsR regulatory elements have been observed in several genes identified from wine LAB. Improved knowledge of regulators and a better understanding of LAB stress responses could constitute a basis of comparison with the well known model microorganisms, Escherichia coli and Bacillus subtilis. Moreover, it can provide an important insight into improving current industrial starter strains.

Bacillus subtilis↗
PubMed2005

Near equiatomic FeCo nanocrystalline alloy embedded in an alumina aerogel matrix: microstructural features and related magnetic properties.

FeCo-Al2O3 nanocomposite aerogels were studied by high-resolution electron microscopy, energy filtered transmission electron microscopy, Mössbauer spectroscopy, and measurements of static magnetizations and hysteretic behavior. The combined use of such techniques provided insights on the formation of bcc FeCo nanocrystalline particles inside the alumina matrix, which is promoted by thermal treatment under hydrogen flow of the parent aerogel. Sample characteristics such as alloy composition and crystallinity, influence of the matrix on the structural evolution, and resulting magnetic properties were investigated as a function of the temperature and time of the reduction treatment.

Journal Article↗
PubMed2005

A beta-glucosidase gene isolated from wine Lactobacillus plantarum is regulated by abiotic stresses.

AIMS: Little genetic information exists on the ability of wine lactic acid bacteria (LAB) to hydrolyse glycoconjugates during malolactic fermentation. We tried to fill this important gap by characterizing a gene codifying for a putative beta-glucosidase enzyme from wine Lactobacillus plantarum and from a commercial strain of Oenococcus oeni. METHODS AND RESULTS: The coding region of the putative beta-glucosidase gene is 1400 nucleotides long and started with an ATG codon. The gene is widespread among LAB and the highest identity was observed between the nucleotide of L. plantarum, Lactobacillus pentosus, Lactobacillus paraplantarum and O. oenibeta-glucosidase gene. The protein sequence deduced from the isolated genes has a calculated molecular mass of 61.19 kDa. Furthermore, the expression of the beta-glucosidase gene in L. plantarum strain was analysed, under several stress, by reverse transcriptase (RT)-PCR and Northern-blot analysis. The gene was apparently regulated by abiotic stresses such as temperature, ethanol and pH. CONCLUSIONS: The beta-glucosidase gene is widespread among LAB and its expression is probably regulated by a wide range of abiotic stresses. SIGNIFICANCE AND IMPACT OF THE STUDY: The inhibitory effect of temperature and ethanol on the L. plantarumbeta-glucosidase gene may be useful to explain the differences found in beta-glucosidase activity reported in wines by several authors.

Amino Acid Sequence↗
PubMed2005

Real-time PCR for the detection of Escherichia coli O157:H7 in dairy and cattle wastewater.

AIMS: Developing and evaluating a rapid real-time polymerase chain reaction (PCR) method for the identification of Escherichia coli O157:H7 in cattle and dairy wastewater samples produced from mozzarella cheese factories, without pre-enrichment step before DNA extraction. METHODS AND RESULTS: Wastewater samples were collected from a dairy farm producing mozzarella cheese and located in Puglia (south of Italy). Plate count and other microbial assays were performed 1 h after sampling. Wastewater samples were artificially inoculated with 10(4), 10(7) and 10(8) cells ml(-1) of E. coli O157:H7, strain EDL933. PCR protocols for stx1, stx2 and eae genes were first tested on pure DNA extracted from type strains, in order to optimize the amplification conditions and reagent concentration before real-time PCR experiments. Three specific fragments of ca 106, 150 and 200 bp corresponding to genes eae, stx1 and stx2, respectively, were obtained. Real-time PCR experiments were performed with DNA extracted from dairy and manure wastewater samples inoculated with 10(4), 10(7) and 10(8) colony-forming units (CFU) ml(-1) of E. coli O157:H7 strain EDL 933. The sensitivity limit of the assay was 10(-1) pg microl(-1) for eae, stx2 and 16SrRNA, and 1 pg microl(-1) for stx1 gene respectively. CONCLUSIONS: A real-time PCR protocol has been developed and used in order to identify potential pathogens in dairy wastewater, in which previous methods (including standard PCR) failed to work. SIGNIFICANCE AND IMPACT OF THE STUDY: Cattle and dairy wastewater samples produced from mozzarella cheese factories may harbour verocytotoxin-producing E. coli. The availability of rapid and sensitive molecular methods may be useful to monitor the persistence of verocytotoxin-producing E. coli in general and to assess the effectiveness of wastewater treatments.

Adhesins, Bacterial↗
PubMed2004

Cloning and characterization of the hsp 18.55 gene, a new member of the small heat shock gene family isolated from wine Lactobacillus plantarum.

Using a molecular approach based on PCR, RT-PCR and northern blot analysis, a new member of the small heat shock family of wine, Lactobacillus plantarum, was cloned and characterized. The protein sequence deduced from the isolated gene had a calculated molecular mass of 18.548 kDa and was therefore named HSP 18.55. The gene codes for a protein homologous to the previously characterized HSP 19.3 and HSP 18.5 and is co-transcribed with an upstream gene of unknown function. Analysis of the 5' flanking region of the hsp 18.55 gene revealed the presence of putative cis elements able to bind alternative sigma factor sigma(B). Based on its structure, the gene was classified as belonging to class II of the heat shock genes according to Bacillus subtilis nomenclature for shock-responsive genes. Expression of the newly identified small heat shock gene, analyzed by RT-PCR and northern blot analysis, was induced by a wide range of abiotic stresses including heat, cold and ethanol, suggesting that the small family of heat shock genes is probably involved in the general stress response in wine L. plantarum. Moreover, the expression of hsp 18.5, hsp 18.55 and hsp 19.3 genes, analyzed over a complete culture cycle, revealed that early growing cells contained substantial amounts of hsp 18.5, hsp 18.55 and hsp 19.3 mRNAs, which rapidly declined upon entry into stationary phase.

Amino Acid Sequence↗
PubMed2004

Molecular characterization of lactic acid populations associated with wine spoilage.

We have investigated the prevalence of spoilage lactic acid bacteria (LAB) in table wines produced in the Apulia region. The occurrence of LAB was evaluated in wines produced with low sulphur dioxide doses and not supplemented with selected malolactic starters such as Oenococcus oeni. About 150 strains were isolated from wine must and a molecular characterization was performed using PCR-based techniques. Most of the strains analysed belonged to Lactobacillus plantarum species. However, some of the strains were identified as Pediococcus damnosus and Leuconostoc sp. The amplified fragments of Pediococcus damnosus were cloned and sequenced. The coding sequence was highly homologous to that of the ropy plasmid confirming that the isolated strain was a ropy(+) Pediococcus damnosus. In all the samples analysed, the final must pH value reached was relatively high (from 3.78 to 3.90). The high pH values had probably negatively influenced (counteracted) the activity of sulphur dioxide added, allowing proliferation of spoilage wine microorganisms.

Bacteria↗
PubMed2004

Expression analysis of putative arcA, arcB and arcC genes partially cloned from Lactobacillus plantarum isolated from wine.

AIMS: The aim of this paper was to study if homofermentative strains (Lacobacillus plantarum) capable of malolactic fermentation in wine can degrade arginine via the ADI pathway. METHODS AND RESULTS: Homofermentative lactic acid bacteria (LAB) isolated from a typical red wine were investigated for their ability to produce citrulline. Citrulline was formed suggesting that the arginine metabolism takes place via the arginine deiminase (ADI) pathway and not via the arginase/urease pathway. Ammonia was also detected with Nessler's reagent, and all the strains examined were able to produce ammonia. Identification of homofermentative LAB was performed using 16S ribosomal sequence analysis. The strains were further classified as belonging to L. plantarum species. Furthermore, the genes encoding for the three pathway enzymes (ADI, ornithine transcarbamylase, carbamate kinase) were partially cloned and gene expression was performed at two different pH values (3.6 and 4.5). CONCLUSIONS: The results suggest that citrulline production in wine, could be performed by homofermentative LAB. SIGNIFICANCE AND IMPACT OF THE STUDY: Homofermentative malolactic bacteria (L. plantarum) may degrade arginine through the ADI pathway.

Amino Acid Sequence↗
PubMed2004

Cloning, molecular characterization and expression analysis of two small heat shock genes isolated from wine Lactobacillus plantarum.

AIM: Understanding the molecular response to stress tolerance of wine Lactobacillus plantarum. METHODS AND RESULTS: Two genes codifying for heat shock proteins were cloned from wine L. plantarum. The coding regions of the two heat shock genes are 420 and 444 nucleotides long, and started with an ATG codon suggesting that they were translated. The protein sequences deduced from the isolated genes have a molecular mass of 18.483 and 19.282 kDa, respectively, and were therefore named hsp18.5 and hsp19.3. The expression of small heat shock genes was analysed by RT-PCR analysis. Moreover, the 5' and 3' noncoding regions were cloned and sequenced. CONCLUSIONS: The expression of the heat shock genes was strongly induced by heat, cold and ethanol stress. Analysis of the 5' and 3' flanking regions of hsp18.5 and hsp19.3 genes, revealed the presence of an inverted repeat sequence (TTAGCACTC-N(9)-GAGTGCTAA) homologue to the CIRCE elements found to the upstream regulatory region of heat shock operons, and an inverted sequence that could form a stem and loop structure that it is likely to function as a transcriptional terminator. Based on their structures, the genes were classified as belonging to Class I of heat shock genes according to the B. subtilis nomenclature of heat response genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Small heat shock genes isolated from wine L. plantarum might have a role in preventing damage by cold stress.

Amino Acid Sequence↗
PubMed2003

Fate of Escherichia coli O157:H7 during the manufacture of Mozzarella cheese.

AIMS: The fate of Escherichia coli O157:H7 was investigated during the manufacture of Mozzarella cheese. METHODS AND RESULTS: The Mozzarella cheese was made from unpasteurized milk which was inoculated to contain ca 10(5) cfu ml(-1)E. coli O157:H7. Two different heating temperatures (70 and 80 degrees C), commonly used during curd stretching, were investigated to determine their effects on the viability of E. coli O157:H7 in Mozzarella cheese. Stretching at 80 degrees C for 5 min resulted in the loss of culturability of E. coli O157:H7 strains, whereas stretching at 70 degrees C reduced the number of culturable E. coli O157:H7 by a factor of 10. CONCLUSIONS: The results show that stretching curd at 80 degrees C for 5 min is effective in controlling E. coli O157:H7 during the production of Mozzarella cheese. Brining and storage at 4 degrees C for 12 h was less effective than the stretching. SIGNIFICANCE AND IMPACT OF THE STUDY: Mozzarella cheese should be free of E. coli O157:H7 only if temperatures higher than or equal to 80 degrees C are used during milk processing.

Cheese↗
PubMed2003

Physiological characterization of 'stay green' mutants in durum wheat.

Four mutants with delayed leaf senescence were selected from seed of durum wheat mutagenized with ethylmethane sulphonate. Changes in net photosynthetic rate, efficiency of photosystem II and chlorophyll concentration during the maturation and senescence of the flag leaves of both mutant and parental plants were determined under glasshouse conditions. The four mutant lines maintained photosynthetic competence for longer than the parental line and are therefore functionally 'stay green'. The mutant lines also had higher seed weights and grain yields per plant than the parental line.

Apoptosis↗
PubMed2002

Characterization of Lactobacillus plantarum from wine must by PCR species-specific and RAPD-PCR.

AIMS: Physiological and molecular analysis such as PCR species-specific and randomly amplified polymorphic PCR (RAPD-PCR) have been used for typing of Lactobacillus plantarum strains from typical wine must. METHODS AND RESULTS: Phenotypic tests such as API 50CH and evaluation of D-L-lactate production from glucose were used to perform a preliminary characterization of lactobacilli. Furthermore, 18 strains of lactobacilli were analyzed by PCR species-specific oligonucleotides based on short sequences of the recA gene. CONCLUSIONS: Four strains were identified as belonging to the L. plantarum species and were further analysed by RAPD-PCR. The RAPD-PCR profiles were similar in all strains that had positive results for species-specific PCR, suggesting that the four L. plantarum strains were closely related. SIGNIFICANCE AND IMPACT OF THE STUDY: Using PCR species-specific as a preliminary screening test and then RAPD-PCR can be as considered the most reliable method of performing a rapid and correct typing of L. plantarum from wine must.

Bacteriocins↗
PubMed1999

Bicalutamide monotherapy versus flutamide plus goserelin in prostate cancer patients: results of an Italian Prostate Cancer Project study.

PURPOSE: To compare the efficacy of bicalutamide monotherapy to maximal androgen blockade (MAB) in the treatment of advanced prostatic cancer. PATIENTS AND METHODS: Previously untreated patients with histologically proven stage C or D disease (American Urological Association Staging System) were randomly allocated to receive either bicalutamide or MAB. After disease progression, patients treated with bicalutamide were assigned to castration. The primary end point for this trial was overall survival. Secondary end points included response to treatment, disease progression, treatment safety, quality-of-life (QOL), and sexual function. RESULTS: A total of 108 patients received bicalutamide and 112 received MAB. There was no difference in the percentage of patients whose prostate-specific antigen returned to normal levels. At the time of the present analysis (median follow-up time, 38 months; range, 1 to 60 months), 129 patients progressed and 89 died. There was no difference in the duration of either progression-free survival or overall survival. However, a survival trend favored bicalutamide in stage C disease but MAB in stage D disease. Overall and subgroup trends were confirmed by multivariate analysis. Serious adverse events and treatment discontinuations were more common in patients receiving MAB (P =.08 and P =.04, respectively). Fewer patients in the bicalutamide group complained of loss of libido (P =. 01) and of erectile dysfunction (P =.002). Significant trends favored bicalutamide-treated patients also with respect to their QOL, namely relative to social functioning, vitality, emotional well-being, and physical capacity. CONCLUSION: Bicalutamide monotherapy yielded comparable results relative to standard treatment with MAB, induced fewer side effects, and produced a better QOL.

Adult↗
PubMed1994

[Semeiotic flowmetry in complex varicocele].

The Authors report on their specific experience on the funicular Doppler veocimetry, regarding the varicocele. In particular we enhance the semeiological spects of the C.W. Doppler velocimetry in the sub-clinical varicocele, in the bilateral one and in the characterization of the spermatic and cremasteric refluxes, and of the external pudenda vein. The Authors stress the utility of the definition of the amount of the reflux, so achieving an integration with the existing classification by degrees, that as it is now days expressed, it seems, to regard the elapsed time and the quantity of reflux caused by the Valsalva manoeuvre. In the practical velocimetry those elements have not been proved to have a parallel increment. Lastly, we report on some cases of veno-spermatic refluxes, uniquely observed in the clynostatism, along with a missing orthostatism. Moreover the Authors believe that some of these hemodynamical consideration could be revisited.

Humans↗
PubMed1987

Cavernous artery revascularization in vasculogenic impotence: new simplified technique.

A new simplified technique of cavernous artery revascularization is presented. The arterial inflow to the corpora is obtained through a counterflow end-to-end anastomosis of the inferior epigastric artery to the proximal stump of the transected dorsal penile artery. The revascularization of the cavernous artery occurs through the common penile artery. This operation is particularly suitable for patients showing pudendal or common penile artery obstruction at arteriography. Optimal distal perfusion is obtained by a cross end-to-side anastomosis of the distal stump of the dorsal artery to the contralateral dorsal penile artery. To date 5 patients have been operated on using this method with good functional results in 80 per cent.

Erectile Dysfunction↗