Antonio Mancini

Significantly improved survival time in pigs with complete liver ischemia treated with a novel bioartificial liver.

Aim of the study was to evaluate treatment efficacy and safety of a scaled-up version of our porcine hepatocytes based BAL system in pigs with complete liver ischemia (LIS). Thirty-one pigs underwent total devascularization of the liver (LIS) by termino-lateral porta-caval shunts and sutures around the bile duct, the common hepatic and gastroduodenal arteries and their accessory branches. The hepato-duodenal ligament was completely transected. Four experimental groups were studied: the first control group (LIS Control, n = 10) received glucose infusion only, the second control group (LIS Plasmapheresis, n = 8) was connected to a centrifugal plasma-separator with a bottle representing the bioreactor volume, the third control group (LIS Empty-BAL, n = 5) received BAL treatment without cells, and the treated group (LIS Cell-BAL, n = 8) was connected for a maximum period of 24 hours to our scaled-up BAL seeded with around 14 billion viable primary porcine hepatocytes. BAL treatment significantly prolonged life in large animals (-35 kg) with complete LIS (Controls, mean ± SEM: 33.1 ± 3 h, Cell-BAL: 51.1 ± 3.4 h; p = 0.001; longest survivor 63 h). In addition, blood ammonia and total bilirubin levels decreased significantly, indicating metabolic activity of porcine hepatocytes in the bioreactor. No significant differences were noticed among the three control groups, indicating that there was no device effect and that the plasmapheresis procedure was well tolerated. No important adverse effectes were observed.

Bridging a patient with acute liver failure to liver transplantation by the AMC-bioartificial liver

Recently a phase I clinical trial has been started in Italy to bridge patients with acute liver failure (ALF) to orthotopic liver transplantation (OLT) by the AMC-bioartificial liver (AMC-BAL). The AMC-BAL is charged with 10 × 109 viable primary porcine hepatocytes isolated from a specified pathogen-free (SPF) pig. Here we report a patient with ALF due to acute HBV infection. This patient was treated for 35 h by two AMC-BAL treatments and was bridged to OLT. There was improvement of biochemical and clinical parameters during the treatment. No severe adverse events were observed during treatment and follow-up of 15 months after hospital discharge. Possible porcine endogenous retrovirus (PERV) activity could not be detected in the patients blood or blood cells up to 12 months after treatment.

Biochemical alterations in semen of varicocele patients: a review of the literature.

Oxidative stress is a mechanism underlying different kinds of infertility in human males. However, different results can be observed in relation to the method used for its evaluation. Varicocele patients show a number of biochemical abnormalities, including an altered distribution of coenzyme Q between seminal plasma and sperm cells and also an apparent defect in the utilization of antioxidants. Moreover, an influence of systemic hormones on seminal antioxidant system was observed too. Finally, the effects of surgical treatment on oxidativestress indexes and the possible usefulness of some medical therapies, like coenzyme Q supplementation, are discussed. In conclusion, published data show a role of oxidative stress in varicocele-related male infertility, but at present we do not know the precise molecular mechanisms underlying these phenomena.

Long‐term absence of porcine endogenous retrovirus infection in chronically immunosuppressed patients after treatment with the porcine cell–based Academic Medical Center bioartificial liver

Di Nicuolo G, D’Alessandro A, Andria B, Scuderi V, Scognamiglio M, Tammaro A, Mancini A, Cozzolino S, Di Florio E, Bracco A, Calise F, Chamuleau RAFM. Long‐term absence of porcine endogenous retrovirus infection in chronically immunosuppressed patients after treatment with the porcine cell–based Academic Medical Center bioartificial liver. Xenotransplantation 2010; 17: 431–439.

Coenzyme Q10 levels in idiopathic and varicocele-associated asthenozoospermia

Levels of coenzyme Q10 (CoQ10) and of its reduced and oxidized forms (ubiquinol, QH2, and ubiquinone, Qox) have been determined in sperm cells and seminal plasma of idiopathic (IDA) and varicocele‐associated (VARA) asthenozoospermic patients and of controls. The results have shown significantly lower levels of coenzyme Q10 and of its reduced form, QH2, in semen samples from patients with asthenospermia; furthermore, the coenzyme Q10 content was mainly associated with spermatozoa. Interestingly, sperm cells from IDA patients exhibited significantly lower levels of CoQ10 and QH2 when compared to VARA ones. The QH2/Qox ratio was significantly lower in sperm cells from IDA patients and in seminal plasma from IDA and VARA patients when compared with the control group. The present data suggest that the QH2/Qox ratio may be an index of oxidative stress and its reduction, a risk factor for semen quality. Therefore, the present data could suggest that sperm cells, characterized by low motility and abnormal morphology, have low levels of coenzyme Q10. As a consequence, they could be less capable in dealing with oxidative stress which could lead to a reduced QH2/Qox ratio. Furthermore, the significantly lower levels of CoQ10 and QH2 levels in sperm cells from IDA patients, when compared to VARA ones, enable us to hypothesize a pathogenetic role of antioxidant impairment, at least as a cofactor, in idiopathic forms of asthenozoospermia.

Thyroid Hormones, Oxidative Stress, and Inflammation.

Inflammation and oxidative stress (OS) are closely related processes, as well exemplified in obesity and cardiovascular diseases. OS is also related to hormonal derangement in a reciprocal way. Among the various hormonal influences that operate on the antioxidant balance, thyroid hormones play particularly important roles, since both hyperthyroidism and hypothyroidism have been shown to be associated with OS in animals and humans. In this context, the nonthyroidal illness syndrome (NTIS) that typically manifests as reduced conversion of thyroxine (T4) to triiodothyronine (T3) in different acute and chronic systemic conditions is still a debated topic. The pathophysiological mechanisms of this syndrome are reviewed, together with the roles of deiodinases, the enzymes responsible for the conversion of T4 to T3, in both physiological and pathological situations. The presence of OS indexes in NTIS supports the hypothesis that it represents a condition of hypothyroidism at the tissue level and not only an adaptive mechanism to diseases.

Effects of Testosterone on Antioxidant Systems in Male Secondary Hypogonadism

Oxidative stress is involved both in metabolic syndrome and male infertility. Hypogonadism is also associated with increased risk for cardiovascular disease. To investigate the role of gonadal steroids in systemic antioxidant regulation, we determined plasma CoenzymeQ(10) (CoQ(10)) and total antioxidant capacity (TAC) in postsurgical hypopituitaric patients. Twenty-six patients aged 28-55 years were studied 6-12 months after surgery. CoQ(10) levels were measured by high-performance liquid chromatography and TAC by spectroscopy with the use of the mioglobin-H(2)O(2) system, which, in interacting with chromogen 2,2(I)-azinobis-(3-ethylbenzothiazoline-6-sulfonate), generates a radical after a latency time (LAG) that is proportional to antioxidant content. Sixteen patients presented low testosterone values; in 10 patients hypogonadism was isolated, and in 6 patients hypothyroidism also was present. CoQ(10) levels were significantly lower in isolated hypogonadism than in normogonadism. Testosterone treatment, performed in those patients with isolated hypogonadism, induced a significant enhancement both in CoQ(10) level and LAG. CoQ(10) and LAG values correlated significantly, suggesting an interrelationship between different antioxidants. Our data suggest that hypogonadism could represent a condition of oxidative stress, in turn related with augmented cardiovascular risk.

Coenzyme Q10 and male infertility

We had previously demonstrated that Coenzyme Q10 [(CoQ10) also commonly called ubiquinone]is present in well-measurable levels in human seminal fluid, where it probably exerts important metabolic and antioxidant functions; seminal CoQ10 concentrations show a direct correlation with seminal parameters (count and motility). Alterations of CoQ10 content were also shown in conditions associated with male infertility, such as asthenozoospermia and varicocele (VAR). The physiological role of this molecule was further clarified by inquiring into its variations in concentrations induced by different medical or surgical procedures used in male infertility treatment. We therefore evaluated CoQ10 concentration and distribution between seminal plasma and spermatozoa in VAR, before and after surgical treatment, and in infertile patients after recombinant human FSH therapy. The effect of CoQ10 on sperm motility and function had been addressed only through some in vitro experiments. In two distinct studies conducted by our group, 22 and 60 patients affected by idiopathic asthenozoospermia were enrolled, respectively. CoQ10 and its reduced form, ubiquinol, increased significantly both in seminal plasma and sperm cells after treatment, as well as spermatozoa motility. A weak linear dependence among the relative variations, at baseline and after treatment, of seminal plasma or intracellular CoQ10, ubiquinol levels and kinetic parameters was found in the treated group. Patients with lower baseline value of motility and CoQ10 levels had a statistically significant higher probability to be responders to the treatment. In conclusion, the exogenous administration of CoQ10 increases both ubiquinone and ubiquinol levels in semen and can be effective in improving sperm kinetic features in patients affected by idiopathic asthenozoospermia

Estradiol Modulation of Basal and Gonadotropin-Releasing Hormone-Induced Gonadotropin Release in Intact and Castrated Men

In 6 intact and 4 castrated adult men a daily intramuscular administration of 1.5 mg estradiol benzoate (E2B) (at 08.00 h) for 7 days induced an initial suppression of circulating gonadotropins with a subsequent rise in LH, which occurred after 96 h of treatment in intact men and after 120 h in castrated men. The magnitude of the LH surge was greater in castrated than in intact men. The changes in basal LH levels were concomitant with a variation in the LH responses to GnRH infusion (0.2 µg/min × 4 h) suggesting a modulatory effect of estrogen on the two pools of pituitary LH. A dramatic decrease in the LH response during the first hour (first pool) was observed after 24-48 h of treatment, followed by a significant increment of this response, which occurred at 96 h in intact men and at 120 h in castrated subjects. During E2B administration, a progressive increment in the delayed response of LH (second pool) was seen, indicating an augmentation of the pituitary capacity. The basal FSH levels progressively decreased and the pattern of FSH response to gonadotropin-releasing hormone (GnRH) infusion was not significantly changed during E2B treatment. The present results demonstrated that:(1) the maintenance for 96-120 h of serum E2 concentrations similar to those present in women at midcycle induced a LH release both in intact and castrated men. (2) The magnitude of LH peak is blunted in the presence of circulating testosterone. (3) E2 induced a biphasic effect on the two pools of pituitary LH similar to that observed in women. Finally, our present data confirm our previous findings that the modulation of gonadotropin secretion in men is not influenced by the perinatal exposure of the hypothalamic-pituitary unit to androgens.

Relationships between Intracranial Pressure and Diurnal Prolactin Secretion in Primary Empty Sella

The role of the intracranial pressure (ICP) in the development and/or maintenance of the primary empty sella has been evaluated by recording the ICP during sleeping and waking periods in 11 women who had this syndrome. Concomitantly, plasma PRL levels, measured at 2-hour intervals during a 24-hour period, were compared with the changes in ICP. Daily PRL variations were also measured in 5 normally cycling and 5 postmenopausal women. ICP was abnormally increased in 8 patients with PES. In 3 of them, increased values were recorded during waking and sleeping periods, while in 5 subjects abnormal values were observed only during sleep. In the remaining 3 patients the ICP was normal in all conditions tested. 8 patients with elevated ICP presented an absent or blunted nocturnal PRL increase. In 3 patients with normal ICP, the circadian periodicity of PRL was preserved. The normalization of ICP obtained in 4 patients by a surgical shunting procedure was accompanied by the return to normal of the circadian PRL periodicity. Our observations demonstrate that the finding of a normal ICP during wakefulness is not sufficient to rule out an actual increase in ICP, since a rise in the CRF pressure can occur during sleep. Our data also demonstrated a correlation between an abnormal rise in ICP and an absent or blunted nocturnal increment in PRL secretion.