Biomedical subjects
Publications and source records attributed to M Carini.
Rapid testicular enlargement is usually strongly suggestive of tumor. Primary testicular infiltration due to amyloidosis is rare. We report a new case of primary testicular amyloidosis in a 72-year-old white man who presented with a short history of painless right testis enlargement.
Liquid Chromatography-Ion Trap Mass Spectrometry with an atmospheric pressure chemical ionization (APCI) interface in the negative and positive-ion modes in parallel to UV-diode-array detection (DAD), was applied for the rapid detection/characterization in crude extracts of the water-soluble antioxidant phenolics from Helichrysum stoechas. APCI-MS provides unequivocal molecular weight data of these compounds and useful information about their structures (diagnostic fragments ions), which were confirmed by the UV-DAD fingerprints. This combined approach allows the identification of ten constituents, including the three naturally occurring isomers of caffeoylquinic acid (CGAs), namely neo-chlorogenic acid, chlorogenic acid and crypto-chlorogenic acid, 2 isomeric dicaffeoyl quinic acids, 2 isomeric naringenin glucosides, quercetin, kaempferol and apigenin glucosides and a tetrahydroxychalcone-glucoside. The water-soluble extract from H. stoechas, standardized in both total polyphenol and kaempferol-3-glucoside content, exhibits strong antioxidant activity in vitro when tested in both artificial membrane systems (phosphatidylcholine liposomes) and in a cell model (rat erythrocytes).
Electron spin resonance (ESR) spectroscopy was applied for the unequivocal detection/quantitation of nitric oxide (NO) as nitrosylhemoglobin (HbFe(II)NO) released from nitroaspirin, benzoic acid,2-(acetyloxy)-3-[(nitrooxy)methyl]phenyl ester (NCX-4016; NO-ASA), the lead of a new class of nonsteroidal anti-inflammatory drugs. In both in vitro and in vivo experiments, the paramagnetic complex was detected at 100 K in the venous blood of the rat (microwave power, 20 mW) and characterized by a three-line hyperfine structure with coupling constants (A(x) and A(z)) of 17 G at g(x)=2.066 and g(z)=2.009. The kinetics of NO release from the drug were first determined in vitro by incubating rat blood with 1 mM NO-ASA and confirmed by the two-line hyperfine structure obtained with the labeled compound ((15)N-NO-ASA). In in vivo studies, the hematic levels of HbFe(II)NO were determined after oral (p.o.) and intraperitoneal (i.p.) administration of the drug (100 and 200 mg kg(-1)). In p.o. treated animals, the complex was detectable at 1 h post-dosing and its formation was maximal at 4-6 h, where the antithrombotic activity peaks. In i.p. treated animals, HbFe(II)NO complex peaks at the second hour to decline thereafter: in these animals, the ESR technique was applied to also detect nitrosylmyoglobin as an index of NO diffusion/compartmentalization in myocardial tissue. The results of this study emphasize the great potentiality of ESR spectroscopy for the study of the release, the metabolic fate and distribution of NO from nitrovasodilators.
The aim of this work was to compare in the rat the cardioprotective efficacy and the total plasma antioxidant activity of a standardised Ginkgo biloba L. extract (GB) as such (300 mg/kg/day) or complexed with phosphatidylcholine (GB-PC; 1:2 w/w), after a 5 days oral administration. At the end of the treatment, the total plasma antioxidant defence was determined by the TRAP and FRAP assays, and the hearts from all groups of animals subjected to moderate ischemia (flow reduction to 1 ml/min for 20 min) and reperfusion (15 ml/min for 30 min). The recovery of left ventricular developed pressure (LVDP) at the end of reperfusion was 35-40% of the preischemic values in both control and vehicle rats, 50.2% in the GB group and 72.5% in the GB-PC pre-treated animals. Creatine kinase (CK) outflow in the perfusate from the hearts of GB and GB-PC treated animals were restrained to a different extent vs. controls (by 71% GB-PC; by 22% GB); the rate of prostacyclin (6-keto-PGF1 alpha) release was far greater in GB-PC than in GB hearts. In parallel, the GB extract significantly increased the total antioxidant plasma capacity (by 24.5% TRAP; 27.9% FRAP) only when complexed with phospholipids. This indicates an increased bioavailability of phenolic antioxidants when suitably embedded within a lipophilic carrier. The results of this study demonstrate that complexation of Ginkgo biloba with phospholipids induces in the rat, even after a short treatment a greater resistance of the heart to ischemia/reperfusion damage in respect to the native extract, due to an increased plasma antioxidant activity.
The inhibitory properties of procyanidins (a standardized oligomeric catechin fraction) from Vitis vinifera L. seeds on the respiratory burst and on the release of granule components myeloperoxidase, beta-glucuronidase and elastase were studied in activated human neutrophils. Procyanidins strongly inhibit superoxide generation with an IC(50) of 7.2 microM, through a direct scavenging of superoxide and prevent the release from calcium ionophore activated neutrophils of beta-glucuronidase (IC(50) = 13.9 microM), myeloperoxidase (IC(50) = 7.2 microM) and elastase (IC(50) = 5.4 microM). In addition they dose-dependently inhibit the activity of myeloperoxidase released from calcium ionophore-stimulated cells with an IC(50) value of 2 microM. The monomeric constitutive unit (+)-catechin was far less active than procyanidins in all the models tested. These results evidence that procyanidins efficiently restrain the inflammatory response of activated neutrophils in vitro and whenever absorbed in vivo can prevent their oxidative discharge at the site(s) of their adhesion.
The QUIBUS study offers some insights into the current diagnosis and treatment of LUTS suggestive of BPH in Italy. As diagnosis is concerned, uroflowmetry and PSA testing were performed in a high percentage of cases (64 and 89%, respectively). Both transrectal ultrasonography and prostate biopsy were more frequent for increasing values of PSA, this suggesting that are used as screening procedures for prostate cancer. However, transrectal ultrasonography was performed overall in a large proportion of patients (61%), representing a routine examination in some centers. As treatment is concerned, the majority of QUIBUS patients had undergone or were undergoing medical therapy. Alphalytics were the drugs most commonly prescribed by urologists while primary care physicians showed the attitude to prescribe more frequently finasteride. On the surgical side, transurethral prostatectomy and open surgery were the most commonly employed procedures, suggesting that little room is left at present to minimally invasive procedures in Italy.
INTRODUCTION: Radical nephrectomy is the treatment of first choice for unilateral renal cell carcinoma (RCC) with a healthy contralateral kidney; however, the current standard for dealing with RCC in patients with a solitary kidney, bilateral tumor and renal or systemic disease inducing a progressive impairment of renal function is nephron-sparing surgery. MATERIALS AND METHODS: Between January 1974 and July 1996, 62 patients (39 men and 23 women, 33-77 years old, mean age 60.6 years) with RCC underwent nephron-sparing surgery. The patients were divided in to two groups according to treatment indication: 46 patients with bilateral tumor (n = 21) or solitary kidney (n = 25) and 16 patients with renal or systemic disease that could damage the contralateral kidney. Survival curves were calculated according to the Kaplan-Meyer method. RESULTS: In the first group 3 patients died postoperatively, and 3 were lost to follow-up; 12 patients (27.9%) had malignant recurrence and 5 (11.6%) died of local recurrence or systemic diffusion. The probability of local or systemic tumor recurrence was 9.9% at 2 years, 20.2% at 5 years and 24.7% at 10 years; the probability of survival was 100% at 2 years, 91.9% at 5 years and 81.9% at 10 years. In the second group 3 patients died of unrelated causes and 1 was lost to follow-up; 4 patients (25%) had a malignant recurrence and 2 (12.5%) died of systemic diffusion of RCC. The probability of tumor recurrence was 13.0% at 2 years, 19.7% at 5 years and 26.4% at 10 years, the probability of survival was 100% at 2 years, 93.3% at 5 years and 86.1% at 10 years. CONCLUSIONS: In our experience nephron-sparing surgery seems justified in patients with a solitary kidney, bilateral tumor or a disease that potentially damages renal function. Tumor diameter and stage, incidental or symptomatic tumor presentation and specific indication for conservative surgery determine the prognosis.
Circulating prostate cells can be detected in peripheral blood of patients with clinically localized or advanced prostate carcinoma. Traditionally, nested reverse transcriptase-polymerase chain reaction (RT-PCR) is used for this as a sensitive, but qualitative only, detection system. We developed a quantitative real-time RT-PCR method for measuring prostate-specific antigen (PSA) mRNA in peripheral blood of prostate cancer patients. A quantitative assay was developed using an external standard reference curve generated with RNA from the human prostate cell line LNCaP. Basal blood samples were collected from 44 patients without evidence of distant metastases and from 30 healthy controls. In 29 patients surgically treated with radical prostatectomy, the measurement of PSA mRNA was performed in blood samples collected before, at the end and 6 days after surgery. In 14 patients treated with radiotherapy, the measurements were repeated at 3-month intervals to evaluate time-related changes during therapy. The measurements were also performed for one year at 3-month intervals in one patient treated with anti-androgen therapy. We found detectable PSA mRNA in 14/44 (32%) basal blood samples. A wide range of values were observed in these patients, ranging from 0.5 to 1724 pg of total LNCaP RNA/ml blood. In patients undergoing radical prostatectomy, circulating PSA mRNA was detectable in eight patients in basal samples, and in seven of them also in blood specimens collected at the end of surgery, showing an increase in only two patients. In blood samples collected 6 days later, PSA mRNA was dramatically reduced in all patients, but still present in seven of them. In four patients, whose basal samples were negative, PSA mRNA was detectable in samples collected at the end of surgery and three of them were negative after 6 days. In patients who did not receive surgical treatment, a rapid decrease in PSA mRNA was demonstrated in five patients treated with radiotherapy and in one patient undergoing androgen deprivation. No detectable PSA mRNA was found in healthy controls. The levels of PSA mRNA in peripheral blood from patients with prostate carcinoma can be easily measured by this sensitive, quantitative and reliable procedure. This assay is a promising tool for the detection and follow-up of these patients.
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When ureteral length is extensively compromised, preservation of the kidney without recurring to external urinary diversion may be attempted by various techniques. The choice depends on the extension and localization of the defect. We report a technique solution performed in a patient with multiple ureteral tumors and solitary kidney. A 70-years old underwent TURB for superficial bladder cancer and right nephroureterectomy for upper urinary tract tumor. He was admitted for hematuria and renal failure which need a percutaneous nephrostomy. Urography and pielo-RMN showed multiple and irregular defects of proximal left ureter. We performed a left ureterectomy with ureteral substitution by tailored and retroperitonealized ileal segment with simultaneous ileal bladder augmentation. The six months follow-up including serum creatinine, sonography, urodynamic evaluation, urinary cytology, excretory urogram and pielo-RMN shows good results. Ureteral replacement with ileum is indicated only for extensive ureteral diseases in which ureteroneocystostomy or vesical Psoas hitch and/or Boari bladder flap are not feasible. With particular attention to surgical aspects as tailoring ileum (to improve propulsion of urinary bolus, limit the absorbing surface area and decrease mucus formation) and the use of an ileal segment longer than 15 cm (to prevent reflux) and with an accurate patient selection, we think that this procedure can assure satisfactory results also in difficult cases.
The persisting shortage of organs for transplantation justifies the use of all possible donors. We compared results for 110 patients, who received solitary kidneys from cadaver donors over age 60 years (OCD) with those for 976 patients, who were transplanted with kidneys from 11-49-year-old cadaver donors, whom we defined as "ideal" age. Although the 4% incidence of primary nonfunction and the 24% rate of delayed graft function were significantly higher (p < 0.001) in the OCD group compared with the ideal group (0.8% and 8.0%, respectively), OCD kidneys can offer good results when low-risk recipients are carefully selected and the cold ischemia time is sharply reduced. Moreover, graft survival rates for 14 OCD grafts, implanted in patients under age 45 were 93% at one, 3 and 10 years compared with 79%, 74% and 42% at the same time points for 96 OCD recipients over age 45. The rate of delayed graft function was higher among 19 OCD grafts preserved for more than 20 hours, and these grafts yielded significantly lower survival rates than 91 OCD grafts preserved for less than 20 hours; with rates of 67%, 58% and 44% and 85%, 81% and 51%, respectively, after one, 3 and 10 years. Thirty-five kidneys from living donors over age 60 had comparable overall graft survival rates to living donor kidneys from donors under age 60 (92%, 92% and 92% vs. 92%, 88% and 80% at one, 3 and 10 years, respectively). An original point scoring system, based only on macroscopic evaluation of the graft, avoids the need for a biopsy and does not prolong cold ischemia time. Microvascular bench reconstructions of the renal artery, damaged by atherosclerosis, expand the possibility for safe transplantation of older kidneys without performing a double renal transplant.
Besides erythema and sunburn reactions, UVB stress can promote erythrocyte extravasation from skin capillaries and hemolysis, and photosensitized hemoglobin can in turn lead to an overload of free radicals in dermis which exacerbates photodamage. The objective of this study was to investigate in rat erythrocytes (RBC) the pattern of events leading to membrane peroxidation and hemolysis following UVB insult (1.5-8.5 J/cm2), and the protective action of grape seed procyanidins. UVB causes a dramatic dose-dependent decrease of intracellular glutathione (paralleled by the formation of pro-oxidant ferryl-hemoglobin), of intramembrane vitamin E and of membrane fluidity, then a rise of conjugated dienes (CD), and thiobarbituric acid-reactive substances (TBARS) and finally a strong hemolytic effect. Procyanidins prevent membrane peroxidation (but not intracellular GSH depletion nor ferryl-hemoglobin formation), with a minimal effective concentration of 0.1 microM (IC50 for TBARS and CD after 120 min UVB exposure: 0.71 microM and 0.56 microM) and dose-dependently delay the onset of hemolysis, by 30 min at 0.1 mciroM, by 90 and 120 min at 0.5 and 1.0 microM. Epigallocatechin-3-O-gallate (EGCG) and catechin, typical constituents of the fraction, were significantly less potent. This since procyanidins (1 microM) inhibit the formation of phospholipid hydroperoxides of the inner (phosphatidylserine, phosphatidylethanolamine) and outer (phosphatidylcholine) layers of the RBC membrane (HPLC analysis), suppress the decrease in membrane fluidity due to lipid and protein thiol oxidation and spare vitamin E from consumption in a dose-dependent manner (0.1-1 microM). Hence procyanidins, preserving membrane phospholipids, since their strong antilipoperoxidant activity, may maintain in vivo the integrity of RBC in sub-epidermal capillaries and effectively counteract in dermis the onset/exacerbation of the UVB-induced skin photodamage.
In this work we describe the development of specific markers for determination of both the membrane and intracellular damage induced by free radicals generated by UVB radiation (5-150 mJ/cm2) in cultured keratinocytes. This using simple, specific and sensitive fluorescent probes: cis-parinaric acid (PNA) to monitor membrane lipid peroxidation and 2',7'-dichloro-dihydrofluorescein diacetate (DCFH-DA) to evaluate the intracellular redox status, in parallel to the fluorimetric determination of the main intracellular antioxidant glutathione. To validate the methodologies, the changes in the intracellular oxidative status following exposure to low doses UVB were measured in both control and N-acetylcysteine-protected cells, in parallel with morphological analyses. UVB induces an early reduction of GSH inside the cell correlated with an increase in the intracellular peroxide content. The effects were time- and dose-dependent. In addition, using a sensitive fluorescent method, we quantitated the release of proteases, a family of proteolytic enzymes greatly involved in the onset/perpetuation of the free radical-induced skin damage from keratinocytes exposed to suberythemal UVB doses (5-15 mJ/cm2). The use of these fluorescent probes provides a reliable tool to detect the early signs of damage in keratinocyte cultures (when the apoptotic phenomenon has not yet been triggered) useful for future screening of protective molecules.
Prostate cancer cells may lose androgen-sensitivity after androgen ablation therapy, becoming highly invasive and metastatic. The biological mechanisms responsible for higher tumurogenicity of androgen-independent prostate carcinomas are not entirely known. We demonstrate that androgen receptor regulation of adhesion and invasion of prostate cancer cells through modulation of alpha6beta4 integrin expression may be one of the molecular mechanisms responsible of this phenomenon. We found that protein and gene expressions of alpha6 and beta4 subunits were strongly reduced in the androgen-sensitive cell line LNCaP respect to the androgen-independent PC3 and that transfection of PC3 cells with a full-length androgen receptor expression vector resulted in a decreased expression of alpha6beta4 integrin, reduced adhesion on laminin, and suppressed Matrigel invasion. Growth in soft agar was also suppressed in androgen receptor-positive PC3 clones. Treatment of androgen receptor positive clones with the synthetic androgen R1881 further reduced alpha6 and beta4 messenger RNA expression as well as adhesion on laminin and Matrigel invasion. Our results indicate that androgens regulate cell-extracellular matrix adhesion and invasion by modulation of integrin expression and function, thus keeping a low invasive phenotype of prostate cancer cells.
Aim of this work was to study the efficacy of procyanidins from Vitis vinifera seeds, a standardized mixture of polyphenol antioxidants, on cardiac mechanics following ischemia/reperfusion stunning in the rat, after 3 weeks supplementation. Young and aged male rats were fed a diet enriched with procyanidins complexed (1:3 w/w) with soybean lecithin (2.4%); control animals (CTR-young and CTR-aged) received an equal amount of lecithin and 2 additional groups of animals the standard diet. At the end of the treatment, the total plasma antioxidant defense (TRAP), vitamin E, ascorbic acid and uric acid were determined in plasma and the hearts from all groups of animals subjected to moderate ischemia (flow reduction to 1 ml/min for 20 min) and reperfusion (15 ml/min for 30 min). In both young and aged rats supplemented with procyanidins the recovery of left ventricular developed pressure (LVDP) at the end of reperfusion was 93% (p < 0.01) and 74% (p < 0.01) of the preischemic values and the values of coronary perfusion pressure (CPP) were maintained close to those of the preischemic period. Also creatine kinase (CK) outflow was restrained to baseline levels, while a 2-fold increase in prostacyclin (6-keto-PGF1alpha) in the perfusate from hearts of young and aged rats was elicited during both ischemia and reperfusion. In parallel, procyanidins significantly increased the total antioxidant plasma capacity (by 40% in young and by 30% in aged rats) and the plasma levels of ascorbic acid, while tend to reduce vitamin E levels; no significant differences were observed in uric acid levels. The results of this study demonstrate that procyanidins supplementation in the rat (young and aged) makes the heart less susceptible to ischemia/reperfusion damage and that this is positively associated to an increase in plasma antioxidant activity.
The aim of this work was to investigate in the rat the protective effect of an oral administration (one week) of Panax ginseng (PG) extract (10 mg/ml in drinking water; 1.6 g/kg/day) on myocardial post-ischemic damage induced by hyperbaric oxygen (HBO) and on the loss in functionality of the endothelium in aorta ring preparations. The hearts from control rats (no-HBO and no-HBO-PG), and from rats exposed to HBO and to HBO after PG treatment were isolated and subjected to mild ischemia and then reperfused. HBO greatly worsens the post-ischemic damage in controls, as demonstrated by the rise of left ventricular end diastolic pressure (LVEDP) and coronary perfusion pressure (CPP). PG significantly restrained the increase of LVEDP and CPP in respect to HBO-untreated rats, as well as that of CPP induced by injection of angiotensin II during pre-ischemia. In HBO control rats the reduction of the vasorelaxant effect of acetylcholine on norepinephrine precontracted aortic rings, was markedly recovered by PG; a similar trend was observed in aortic rings challenged with the nitric oxide synthase inhibitor NG-monomethyl-L-arginine (56% recovery). These results strongly indicate that PG prevents the myocardial ischemia/reperfusion damage and the impairment of endothelial functionality induced by reactive oxygen species arising from HBO exposure, through an antioxidant intervention. The in vitro radical scavenging activity of PG seems to be too weak (0.05-0.5 mg/ml) to explain by itself the cardiac and extra-cardiac protective effects, and this suggests a role also for an indirect antioxidant action of the drug (endothelial nitric oxide synthase stimulation).
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