Synthesis and biological assessment of 3,7-dihydroxytropolones.

3,7-Dihydroxytropolones (3,7-dHTs) are highly oxygenated troponoids that have been identified as lead compounds for several human diseases. To date, structure-function studies on these molecules have been limited due to a scarcity of synthetic methods for their preparation. New synthetic strategies towards structurally novel 3,7-dHTs would be valuable in further studying their therapeutic potential. Here we describe the successful adaptation of a [5 + 2] oxidopyrilium cycloaddition/ring-opening for 3,7-dHT synthesis, which we apply in the synthesis of a plausible biosynthetic intermediate to the natural products puberulic and puberulonic acid. We have also tested these new compounds in several biological assays related to human immunodeficiency virus (HIV), hepatitis B virus (HBV) and herpes simplex virus (HSV) in order to gain insight into structure-functional analysis related to antiviral troponoid development.

Cost-benefit evaluation of a preventive intervention on the biological risk in health: the accidental puncture during the administration of insulin in the University Hospital "Federico II" of Naples.

BACKGROUND: The occupational exposure to biological risk is a frequent event that affects millions of workers in the health sector. Operators are exposed to accidental contact with blood and other potentially infectious biological materials with a frequency higher than that observed in the population (occupational exposure). The pathogens most frequently implicated are the human immunodeficiency virus (HIV), hepatitis C (HCV) and hepatitis B (HBV) viruses. The World Health Organization estimates that each year more than 3 million health workers hurt themselves with an object/edge definitely contaminated with at least one HIV (about 170,000 exposures), hepatitis B (approximately 2,000,000 exposures) and hepatitis C (approximately 900,000 exposures). In Italy approximately 100,000 percutaneous exposures/year are estimated to take place. The needlestick injuries in health care workers are, in large part, preventable by adopting measures such as the use of instrumental needlesticks Prevention Devices - NPDs. The adoption of the NPDs is extremely effective in reducing occupational exposure to biological risk (from 63 % to 100 % reduction). METHODS: The aim of this study was to evaluate whether the adoption of NPDs for insulin therapy is costeffective in terms of prevention of accidents by Biohazard, compared to administration of insulin with traditional methods (syringe + vial). The estimation is carried out both in the light of current legislation (European Directive 2010/32 and 81/08 Italian Law) and epidemiological data and cost of accidents (according to frequency) and alternative interventions. RESULTS: The evaluation of cost-effectiveness included the construction of an economic model that would allow the weighting of the costs of accidents that can occur following the administration of insulin therapy with traditional methods. The economic model was developed taking into account the international literature on the phenomenon of "accidental puncture" and allowed the financial quantification of the event. Then we calculated the cost of insulin therapy using the traditional methodology and the cost has been converted to the cost of insulin therapy when administered by NPDs. The period of the study was the year 2010. CONCLUSIONS: The data thus obtained were used to evaluate the benefits of implementing NPDs for insulin therapy, in terms not only of economic advantage but also of preventive efficacy and on the cost of the accident.

[Surgical site infections in Italy]. / Le infezioni del sito chirurgico in Italia.

Surgical site infections (SSIs) are recognized as a common surgical complication, occurring in about 2-5% of all surgical procedures. SSIs represent the third most frequent nosocomial infection, accounting for 14.6% of all infections observed in hospitalised patients and 38% of those observed among surgical patients. Strategies for the prevention of SSIs also include surveillance which has proved very effective. The most recent surveillance study carried out at a national level in Italy is Kir-Nos, a multicentric study sponsored by GlaxoSmithKline and performed between April and June 2002 in 32 different General Surgery Units for a total of 2972 surgical patients enrolled. Results emerging from the study clearly indicate that many patients receive inappropriate antimicrobial prophylaxis, especially in terms of drug choice, route and timing of administration. Given the high economic burden that infections provoke, beyond the increased morbidity and mortality, it appears mandatory to improve our tools in order to reduce their incidence, as a reduction of only 0.1% can result in a considerable saving of economic resources to be allocated to other activities, such as screening and prevention programs.

alpha1-adrenergic receptor subtypes in human peripheral blood lymphocytes.

We investigated the expression of alpha1-adrenergic receptor subtypes in intact human peripheral blood lymphocytes using reverse transcription-polymerase chain reaction (RT-PCR) and radioligand binding assay techniques combined with antibodies against the three subtypes of alpha1-adrenergic receptors (alpha1A, alpha1B, and alpha1D). RT-PCR amplified in peripheral blood lymphocytes a 348-bp alpha1A-adrenergic receptor fragment, a 689-bp alpha1B-adrenergic receptor fragment, and a 540-bp alpha1D-adrenergic receptor fragment. Radioligand binding assay with [3H]prazosin as radioligand revealed a high-affinity binding with a dissociation constant value of 0. 65+/-0.05 nmol/L and a maximum density of binding sites of 175. 3+/-20.5 fmol/10(6) cells. The pharmacological profile of [3H]prazosin binding to human peripheral blood lymphocytes was consistent with the labeling of alpha1-adrenergic receptors. Antibodies against alpha1A-, alpha1B-, and alpha1D-receptor subtypes decreased [3H]prazosin binding to a different extent. This indicates that human peripheral blood lymphocytes express the three alpha1-adrenergic receptor subtypes. Of the three different alpha1-adrenergic receptor subtypes, the alpha1B is the most represented and the alpha1D, the least. Future studies should clarify the functional relevance of alpha1-adrenergic receptors expressed by peripheral blood lymphocytes. The identification of these sites may represent a step for evaluating whether they represent a marker of alpha1-adrenergic receptors in cardiovascular disorders or for assessing responses to drug treatment on these receptors.

Impaired cortisol binding to glucocorticoid receptors in hypertensive patients.

We compared glucocorticoid receptor binding characteristics and glucocorticoid responsiveness of human mononuclear leukocytes (HML) from hypertensive patients and matched normotensive volunteers. We also considered associations of these variables with plasma renin activity, aldosterone, cortisol, corticotropin, and electrolyte concentrations. We calculated binding affinity (Kd; nmol/L) and capacity (Bmax; sites/cell) for dexamethasone and cortisol from homologous and heterologous competition curves for specific [3H]dexamethasone binding sites on HML isolated from the blood of normotensive volunteers and subjects with essential hypertension. Glucocorticoid responsiveness of HML was evaluated as IC50 values (nmol/L) for dexamethasone and cortisol for the inhibition of lysozyme release. We measured plasma hormones by radioimmunoassay. Kd values (mean+/-SE) for cortisol in HML of hypertensive patients were higher than in control subjects (24.6+/-2.4 versus 17.5+/-1.7 nmol/L, P<.04). Binding capacity (4978+/-391 versus 4131+/-321 sites/cell), Kd values for dexamethasone (6.7+/-0.5 versus 5.7+/-0.3 nmol/L), and IC50 values for dexamethasone (3.4+/-0.3 versus 3.1+/-0.2 nmol/L) and cortisol (12.2+/-1.6 versus 9.5+/-0.3 nmol/L) were not significantly different. Patients with renin values less than 0.13 ng angiotensin I/L per second were markedly less sensitive to cortisol than those with higher values. Both Kd (30.3+/-2.5 versus 19.2+/-2.4 nmol/L) and IC50 values (15.5+/-1.8 versus 8.9+/-1.2 nmol/L) for cortisol were significantly higher in patients with lower renin values (P<.03). Other variables, including plasma hormone and electrolyte values and binding characteristics for dexamethasone, were not different. These data suggest that cortisol binding to glucocorticoid receptor is slightly impaired in patients with essential hypertension. In vivo, this could lead to inappropriate binding of cortisol to mineralocorticoid receptors. Hence, decreased sensitivity to cortisol is associated with renin suppression. This hypothesis is supported by evidence of hypertension and low renin activity, which others have described in patients with primary glucocorticoid resistance due to mutations of the glucocorticoid receptor.

Angiotensin-converting enzyme and angiotensinogen gene polymorphisms are non-randomly distributed in oral contraceptive-induced hypertension.

OBJECTIVES AND METHODS: Oral contraceptives (OC) usage increases serum angiotensinogen levels to three to five times normal and about 5% of these women develop arterial hypertension. The genetic contribution to this susceptibility to OC-induced hypertension is poorly understood. We have analyzed the genotypes of 149 hypertensive and 101 normotensive women using oral contraceptives, for three genetic polymorphisms in genes of the renin-angiotensin system: an insertion/deletion (I/ D) in the angiotensin converting enzyme (ACE) gene, the T235M polymorphism of the angiotensinogen gene (AGT) and a point mutation in its promoter. RESULTS: After cessation of oral contraception the mean arterial pressures of the hypertensive women were separable into two non-overlapping groups; 88 of the women remained hypertensive and 61 returned to normal blood pressure. Both groups of hypertensive women had a similarly higher frequency of hypertensive relatives than the normotensive women, but were otherwise similar. The 235T allele of AGT was significantly increased in frequency in the 61 oral contraceptive-inducible hypertensive women compared with the controls and the 88 women that remained hypertensive. The ACE I/D genotypes were similarly distributed within the three groups of women, but were distinctly non-random in the oral contraceptive-induced hypertensive women when they were also classified by AGT genotype. CONCLUSION: This statistical interaction of genotype frequencies suggests that the genetic basis of susceptibility to OC-induced hypertension is complex.

Alpha1-adrenergic receptor subtypes in peripheral blood lymphocytes of essential hypertensives.

OBJECTIVE: The expression of alpha1-adrenergic receptor subtypes in peripheral blood lymphocytes was investigated in 28 essential hypertensive patients as well as in the peripheral blood lymphocytes and aorta of spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. METHODS: Alpha1-adrenergic receptors were quantified by radioligand binding assays, employing [3H]-prazosin as the radioligand in association with compounds displaying different degrees of selectivity for alpha1A-, alpha1B- and alpha1D-adrenergic receptor subtypes. RESULTS: The affinity of [3H]-prazosin binding was similar in peripheral blood lymphocytes of different stage essential hypertensive and normotensive subjects or of SHR and age-matched normotensive WKY rats as well as in the aortas of SHR and WKY rats. The radioligand binding assay revealed no change in the expression of alpha1-adrenergic receptors in peripheral blood lymphocytes of essential hypertensives compared with normotensive subjects; a moderate decrease of alpha1B-adrenergic receptors and an increase of alpha1D-adrenergic receptors. The relative densities of the alpha1-adrenergic receptor subtypes were similar in the three groups of essential hypertensives. In peripheral blood lymphocytes and in aorta of SHR, [3H]-prazosin binding was significantly reduced compared with normotensive WKY rats. The expression of alpha1-adrenergic receptor subtypes in peripheral blood lymphocytes of SHR was similar to that found in peripheral blood lymphocytes of essential hypertensives. CONCLUSIONS: Changes of lymphocyte alpha1-adrenergic receptor subtypes in essential hypertensives are similar to those observed in lymphocytes and vascular tissues of animal models of hypertension. This suggests that assays of lymphocyte alpha1-adrenergic receptors may represent an indirect marker of their involvement in essential hypertension.

Vasoactive hormones induce nitric oxide synthase mRNA expression and nitric oxide production in human endothelial cells and monocytes.

Isoform-2 nitric oxide synthase (NOS-2) mRNA expression and nitric oxide (NO) production are induced in endothelial cells and monocytes by cytokines such as gammaIFN and LPS. We evaluated NOS-2 and isoform-3 NOS (NOS-3) mRNA expression and NO production in human monocytes and human umbilical vein endothelial cells (HUVEC), under basal conditions and after incubation with physiologic concentrations of vasoactive hormones. NOS mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and NO production by electronic paramagnetic resonance spectroscopy (EPR). We showed that NOS-2 mRNA expression and NO production were induced by stimulation with epinephrine, dopamine, endothelin-1, and angiotensin II, both in monocytes and HUVEC. NOS-3 mRNA expression and NO production were detected under basal conditions in monocytes and HUVEC and were not modified by the presence of vasoactive hormones. Human endothelial cells and monocytes express the NOS-2 and NOS-3 mRNA and the inducible NOS-2 mRNA expression increases after vasoactive hormone stimulation.

Blood pressure and heart rate in young thalassemia major patients.

The analysis of blood pressure (BP) and heart rate (HR) variability is currently used to investigate the mechanisms responsible for cardiovascular control; therefore, we assessed whether an impairment of 24-h BP and HR profiles and sympathovagal interaction modulating cardiovascular function was present in patients with thalassemia major (TM) in preclinical phase of heart disease. Nine beta-thalassemic patients 18 years old without clinical signs of cardiac failure and 9 age- and sex-matched controls were studied. Twenty-four-hour-ambulatory BP and HR were measured using the SpaceLabs 90207 device. A truncated Fourier series with four harmonics was used to describe the diurnal blood pressure profile. Mean 24-h ambulatory systolic BP, diastolic BP, and mean arterial pressure were significantly lower in TM patients than in normal subjects (P < .05). A significantly higher nighttime HR value was found in TM patients (P < .05). More than 40% of the TM patients did not show a significant diurnal BP and HR rhythm. In TM patients, the overall amplitude of systolic BP, diastolic BP, and HR was significantly lower than in controls (P < .01). The night/day differences of systolic BP, diastolic BP, and HR were significantly lower in TM patients than in normals (P < .01). Furthermore, we performed power spectral analysis on short-term continuous finger BP and HR data in supine position and during passive head-up tilt. Total spectral power of systolic BP was significantly lower in patients than controls (P < .05). Low-frequency (LF) power of systolic BP and diastolic BP and LF/high-frequency (HF) ratio of HR were significantly lower during tilt in TM patients compared to controls (P < .05). High-frequency power of HR was significantly higher in patients than controls (P < .05). The baroreflex gain assessed by alpha-index was the same in supine position but was higher in TM patients during passive tilt (P < .05). An inverse relationship between LF/HF ratio of HR and hemoglobin levels in TM patients was found. Finally, plasma norepinephrine levels were significantly lower in thalassemics (P < .005). In young TM patients in a preclinical stage of heart disease, these findings demonstrated abnormal 24-h BP and HR rhythms and a decreased short-term variability of BP and HR, in particular in the LF range, showing a diminished sympathetic activity.

A nosocomial outbreak of Serratia marcescens producing inducible Amp C-type beta-lactamase enzyme and carrying antimicrobial resistance genes within a class 1 integron.

We investigated an outbreak of Serratia marcescens in the adult intensive care unit of the University Hospital of Napoli. The outbreak involved 13 cases of infection by S. marcescens over a nine-month period and was caused by a single pulsed-field gel electrophoresis clone. The epidemic strain was multiply antibiotic resistant, producing an inducible Amp C-type beta-lactamase enzyme and carrying the trimethoprim-resistance gene and the adenyltransferase gene, which confers resistance to streptomycin and spectinomycin, within a class 1 integron. Antimicrobial therapy with beta-lactams was associated with S. marcescens acquisition in the intensive care unit.

[The L-arginine/nitric oxide metabolic pathway. Its physiopathology and clinical implications]. / La via metabolica L-arginina/ossido di azoto. Fisiopatologia ed implicazioni cliniche.

The L-arginine/nitric oxide (NO) pathway plays a key role in a number of biological processes within most organs and systems. Increasing attention has been addressed to its involvement in the pathogenesis of various human diseases. In this review we examine the enzymology of different NO-synthase isoforms, the major NO detection techniques as well as the possible clinical and pharmacological implications of this new metabolic pathway.

Diagnostic value of urinary cyclic AMP measurement in patients with calcium nephrolithiasis.

One hundred patients with recurrent calcium nephrolithiasis were submitted to the Pak test. At fasting state hypercalciuria was found in 27 cases, while a group of 16 further patients became hypercalciuric after oral calcium load. Only measurement of urinary cAMP excretion in both conditions made it possible to diagnose renal hypercalciuria in 9 out of 27 patients in the former group; according to test results 4 patients were expected to have primary hyperparathyroidism, but afterwards the disease was identified in only one case.

[Histopathologic risk factors in patients with non-seminomatous germ tumors of the testis in clinical stage 1. Retrospective study of 75 patients]. / Fattori istopatologici di rischio in pazienti con tumore germinale non seminomatoso del testicolo in stadio clinico 1. Uno studio retrospettivo su 75 pazienti.

OBJECTIVES: This retrospective study was performed to evaluate histopathologic prognostic risk factors in 75 patients on clinical stage 1 nonseminomatous germ cell cancer of the testis (NSGCTT). METHODS: From September 1976 to February 2000 we operated on 75 patients for NSGCTT on clinical stage 1 disease. Average age was 29.5 years (range 16-71). After orchiectomy, therapeutic options included retroperitoneal lymph node dissection (RLND) for 44 patients (58.6%), surveillance for 26 (34.6%) and neoadjuvant chemotherapy for 5 (6.6%). Testis primary tumor samples were assessed for studying prognostic risk factors that included vascular and/or lymphatic invasion (IV/IL+), percentage of embryonal carcinoma (%EC) and absence of yolk sac tumor (YS-). RESULTS: All patients were alive and disease-free. The average age follow-up was 84.5 months (range 1-254). Relapses occurred in 11 (14.6%) patients after an average follow-up of 9.09 months (range 3-24). Prognostic risk factors were detected as follows: IV/IL+ in 17 cases (22.7%), (50-80%) EC in 23 (30.6%), CE% > 80 in 23 (30.6%), YS- in 55 (72%). In 8 (10.6%) patients there was not any prognostic risk factor. Disease relapse related to prognostic risk factors was detected as follows: 18.1% for VI/LI, 90.9% for EC% > 50 (27.2% for 50-80% EC and 63.6% for CE% > 80) and 90.9% for YS-. Relapsing rates between patients with EC% > 80 and 50-80% EC resulted statistically significant (p = 0.02, odds ratio = 12.25). Relapsing rates between patients on surveillance and those who underwent RLND was next to be significant (p = 0.05, odds ratio 3.68). CONCLUSIONS: EC% > 80 is a prognostic risk factor for disease relapse in patients with clinical stage 1 NSGCT who are selected in a high risk group requiring RPLND or neoadjuvant chemotherapy as therapeutical option.

[Inverted papilloma of the bladder (considerations on a clinical case )]. / Il papilloma invertito della vescica (considerazioni su un caso clinico).

The Authors present one case of inverted papilloma of the bladder. Morphology, clinical features and prognosis of this lesion are discussed.

[Cutaneous urostomies: indications, technics, treatments]. / Urostomie cutanee: indicazioni, tecniche, cure.

The authors study the outer urinary derivations; place the indications to perform urostomies on the basis of the possible pathological situations; describe the technical modalities for every outer derivative operation concerning kidneys, ureters, vesica and urethra, respectively. They, at last, consider the eventual complications of such operations, and report about the therapeutical and prophylactic measures to be taken, with particular attention to the urine collecting devices.

[The complications of lumbotomies]. / Le complicazioni delle lombotomie.

After having described the surgical anatomy of the lumbar region, with particular regard to the relationships with pleura and peritoneum, as well as to the course of the nervous structures of the region (12th intercostal, ileohypogastric and ileoinguinal), the authors describe the possible complications related with the surgical approaches to the lumbar region. Pathogenesis of post-operatory pains and parietal hypotonies is especially evaluated; these complications may be caused by section, ligature or compression of one or more nervous trunks. The authors conclude that the respect of the anatomic structures located in the lumbar region represents the best way in order to prevent most complications related to lumbotomies.

[Comparison of the quality of life of patients treated by surgery or radiotherapy in epidermoid cancer of the penis]. / Comparaison de la qualité de vie des patients traités pour cancer épidermoïde de la verge par chirurgie ou radiothérapie.

OBJECTIVE: The purpose of our work is to examine how partially or totally radical surgery and radiotherapy can weigh on quality of life in patients with squamous carcinoma of the penis, considering whether quality of life can be a good criterion in treatment options. MATERIAL AND METHODS: We have interviewed 17 patients alive and disease-free (average follow-up 69.43 months) after surgical treatment or radiotherapy for cancer of the penis (emasculation, 2; total amputation of the penis, 2; partial penectomy, 11; radiotherapy, 2). Tests used: General Health Questionnaire (G.H.Q.-12 by D. Goldberg); Hospital Anxiety and Depression Scale (H.A.D.S.); Social Problem Questionnaire (S.P.Q.); Overall Sexual Functioning Questionnaire (O.S.F.Q); Family APGAR Questionnaire (F.A.Q.); Performance Status ECOG. RESULTS: Thirty-five per cent of the patients presented limitations in their state of health and social problems. Anxiety was evident in 29.5% of the patients and depression in 6%. The global sexual function was compromised in 76.5%. CONCLUSION: The profile of quality of life resulted compromised in the patients who had undergone radical surgical treatment. The sexual function was the most altered component. The impact of the pathology and its treatment on the other domains of quality of life proved less significant.