Genetic polymorphisms of Fas (CD95) and FasL (CD178) in human longevity: studies on centenarians.

Apoptosis plays a crucial role in immunosenescence, as also evidenced by the increased expression of Fas in lymphocytes from aged people. However, little is known about the genetic regulation of Fas and its ligand, FasL. We have studied their polymorphisms in 50 centenarians and 86 young donors living in Northern Italy. The first Fas polymorphism, at position -670, has in Caucasian a heterozigosity of 51%; the second, at -1377 position, has the wild type allele (G) with a very high frequency (83%) respect to the mutant allele. Genotype and allele distribution for both polymorphisms were similar in controls and centenarians. Similar results were found as far as two FasL polymorphisms (IVS2nt-124 and IVS3nt169) are concerned. On the whole, our data suggest that Fas and FasL polymorphisms, as well as their haplotypes, are unlikely to be associated with successful human longevity.

Apoptosis-resistant phenotype in HL-60-derived cells HCW-2 is related to changes in expression of stress-induced proteins that impact on redox status and mitochondrial metabolism.

The onset of resistance to drug-induced apoptosis of tumour cells is a major problem in cancer therapy. We studied a drug-selected clone of promyelocytic HL-60 cells, called HCW-2, which display a complex resistance to a wide variety of apoptosis-inducing agents and we found that these cells show a dramatic increase in the expression of heat shock proteins (Hsps) 70 and 27, while the parental cell line does not. It is known that stress proteins such as Hsps can confer resistance to a variety of damaging agents other than heat shock, such as TNF-alpha, monocyte-induced cytotoxicity, and also play a role in resistance to chemotherapy. This elevated expression of Hsps is paralleled by an increased activity of mitochondrial metabolism and pentose phosphate pathway, this latter leading to high levels of glucose-6-phosphate dehydrogenase and, consequently, of glutathione. Thus, the apoptotic-deficient phenotype is likely because of the presence of high levels of stress response proteins and GSH, which may confer resistance to apoptotic agents, including chemotherapy drugs. Moreover, the fact that in HCW-2 cells Hsp70 are mainly localised in mitochondria may account for the increased performances of mitochondrial metabolism. These observations could have some implications for the therapy of cancer, and for the design of combined strategies that act on antioxidant defences of the neoplastic cell.

Changes in intramitochondrial cardiolipin distribution in apoptosis-resistant HCW-2 cells, derived from the human promyelocytic leukemia HL-60.

Using a cytofluorimetric approach, we studied intramitochondrial cardiolipin (CL) distribution in HCW-2 cells, an apoptosis-resistant clone of human HL-60 cells. In HL-60, about 50% of total CL is distributed in the outer leaflet of mitochondrial inner membrane, while in HCW-2 a significantly higher amount of CL (about 65%) is in that site. In basal conditions, HSW-2 cells also show a reduced mitochondrial membrane potential even if they are able to proliferate as the parental line. Taking into account the complex functions that CL plays in the regulation of mitochondrial activity, it is likely that HCW-2 could produce ATP utilizing more glycolytic pathways rather than mitochondrial respiratory chain.

Plasma antioxidants and longevity: a study on healthy centenarians.

A large body of experimental research indicates that oxidative stress contributes to the processes related to aging and to the pathogenesis of several age-related diseases. Vitamins and antioxidant enzymes have a fundamental role in defending the organism from oxidative stress. To better understand the role of antioxidants in human aging, we measured plasma levels of vitamin C (ascorbic acid), uric acid, vitamin E (alpha-tocopherol), vitamin A (retinol), carotenoids, total thiol groups, and the activity of plasma superoxide dismutase (SOD) and glutathione peroxidase (GPX) as well as the activity of red blood cell (RBC) SOD in 32 healthy centenarians-17 elderly subjects aged 80-99 years, 34 elderly subjects aged 60-79 years, and 24 adults aged less than 60 years. Considering the "noncentenarians" only, we observed a consistent behavior in the antioxidant pattern, with a decrease of the nonenzymatic antioxidants and an increase of the enzymatic antioxidant activities relative to age. Remarkably, centenarians were characterized as having the highest levels of vitamins A and E, whereas the activities of both plasma and RBC SOD, which increase with age, decreased in centenarians. From these results, it is evident that healthy centenarians show a particular profile in which high levels of vitamin A and vitamin E seem to be important in guaranteeing their extreme longevity.

Apoptosis--programmed cell death: a role in the aging process?

Cells continuously exposed to genotoxic agents, such as oxygen free radicals (OFRs), deeply involved in the aging process use a variety of cellular defense mechanisms. These defense mechanisms include DNA repair enzymes, antioxidants, poly(ADP-ribosyl)polymerase (pADPRP), and stress proteins and they constitute an integrated network. An age-related failure of the efficiency of this network can affect cell proliferation and cell death, two phenomena tightly linked and regulated. Recent data from our laboratory on the role of DNA damage and pADPRP activation and on the type of cell death induced by OFRs in human lymphocytes are reviewed. In vitro and in vivo data on possible strategies to reduce oxidative stress in lymphocytes from normal and Down syndrome subjects, by using natural compounds and trace elements, are presented. They indicate that nicotinamide and L-carnitine protect human cells from OFR-induced damage and suggest that they are possible candidates as antiaging substances.

A cytofluorimetric study of T lymphocyte subsets in rat lymphoid tissues (thymus, lymph nodes) and peripheral blood: a continuous remodelling during the first year of life.

We previously demonstrated that the rat thymus undergoes a progressive remodelling long before the appearance of typical signs of involution [Quaglino, D., Capri, M., Bergamini, G., Euclidi, E., Zecca, L., Franceschi, C., Pasquali Ronchetti, I., 1998. Age-dependent remodelling of rat thymus. Morphological and cytofluorimetric analysis from birth up to one year of age. Eur. J. Cell. Biol. 76, 156-166]. To focus better on the complex remodelling that occurs in the rat immune system during the first year of life, we analysed the phenotype profile of thymocytes, and T lymphocytes from mesenteric lymph nodes and peripheral blood of the same animals by flow cytometry. Two experimental sets were performed simultaneously using the same animal strain, but starting and ending the study at different ages (15 days up to 300 days in the first experimental set, and 90 days up to 360 days of life in the second). In the rat these ages appear to be crucial not only for developmental, maturative and early involutional processes of the thymus, but also of the entire immune system. The main findings were the following: (1) in the thymus, CD8(-)CD4(-) cells increased, CD5(+)alphabeta TCR(-) and CD8(+)CD4(+) thymocytes decreased, while the most mature cell subset appeared well preserved with ageing; (2) in the lymph nodes, T helper and T cytotoxic lymphocytes decreased in the most aged animals. Memory/activated CD4(+)CD45RC(-) T cells decreased, while naive/resting CD4(+)CD45RC(+) cells increased in the youngest animals and decreased in the oldest. CD8(+)CD45RC(-) and CD8(+)CD45RC(+) lymphocytes showed a complex age-dependent trend, and (3) in peripheral blood, minor modifications were evident, such as an age-dependent increase in the alphabeta TCR(+)CD25(+) cell subset. Some of these changes were related to the developmental process, while others could likely be interpreted as early signs of immunosenescence. The role of these modifications in immune system is discussed within the framework of the remodelling hypothesis of immunosenescence. The age-dependent changes in these three lymphoid compartments, however, appear to be different and only partially overlapping, thus suggesting that the maturational, developmental and ageing processes have distinct characteristics in the central and peripheral lymphoid organs.

Mitochondria and HIV infection: the first decade.

In the last few years, the interactions between mitochondria and infection with the human immunodeficiency virus (HIV) have received careful attention. Starting from the first studies regarding the presence of mitochondrial damage in cardiac tissue from patients who died of AIDS, researchers have investigated different aspects of the interactions between the virus and mitochondria, from acute primary infection to the final stages of the disease. Only recently a significant impulse to this type of research has come from the observation that the so called "highly active antiretroviral therapy" (HAART), a combination of potent antiretroviral drugs such as viral protease inhibitors or nucleoside-analogue reverse-transcriptase inhibitors, is capable of damaging these organelles and cause a clinical syndrome called lipodystrophy. There is still an open debate concerning the exact responsibility of HAART as well as on metabolic pathways and mechanisms that are involved in the onset of lipodystrophy. The hypothesis that drug-induced damage to mitochondrial (mt) DNA is able to alter mitochondria functionality to a similar extent as that occurring in genetic disease affecting mtDNA suggests that mitochondria plays a crucial role in the pathogenesis of this syndrome. In this paper, data concerning the interactions between mitochondria and HIV infection will be reviewed.

Hepatoma HepG2 cells as a model for in vitro studies on mitochondrial toxicity of antiviral drugs: which correlation with the patient?

Currently, drugs have been synthesised that can significantly delay the course of several viral infections, including those provoked by HBV, HCV or HIV, but that display consistent side effects, including toxicity for organelles such as mitochondria. Several in vitro models and techniques have been developed to analyse the effects of such compounds. HepG2 cells (from human hepatoma) are an excellent model to investigate mitochondrial (mt) toxicity because of their high content of organelles and mtDNA, and actually different investigators are indeed using such cells. Studies in vitro on cell lines are relatively easy, but it is necessary to be careful in the interpretation of data, which are usually obtained on continuously growing, tumour cells, quite different from normal, resting, non-neoplastic cells collected from a patient. Direct analysis of drug-induced mt damage in patients is extremely more complex than that performed using in vitro models because of the difficulty to obtain adequate cells or to have discrete amounts of biological material, the status of the patient at the moment of cell collection, the use of an adequate assay and its correct execution, and finally the possibility to find sex- and age-matched healthy controls as source of reference parameters.

Estroprogestin vs. gonadotrophin agonists plus estroprogestin in the treatment of endometriosis-related pelvic pain: a randomized trial. Gruppo Italiano per lo Studio dell'Endometriosi.

OBJECTIVE: This is a randomized clinical trial comparing estroprogestin (E/P) pill given for 12 months vs. gonadotrophin releasing hormone agonist (GNRHa) given for 4 months followed by E/P pill treatment for 8 months in the relief of endometriosis-related pelvic pain. METHODS: Eligible for the study were women with laparoscopically confirmed endometriosis and pelvic pain lasting 3-12 months after diagnosis. Eligible women were randomly assigned to treatment with E/P pill (gestroden 0.75 mg and ethynlestradiol 0.03 mg) for 12 months (47 patients) vs. tryptorelin 3.75 mg slow release every 28 days for 4 months followed by E/P pill for 8 months (55 patients). RESULTS: At baseline, dysmenorrhea was reported in 46 women allocated to E/P pill only (97.9%), and in all the 55 women allocated to GNRHa+E/P pill. The corresponding value at the 12 months follow-up visit was 14 subjects (35.9%) and 16 subjects (34.8%). The baseline median values of the multidimensional and analog scale were for dysmenorrhea 4 and 6 in the EP only and 3 and 6 in the GNRHa+E/P group. The corresponding value at the 12 months follow-up visit were 2 and 6 and 0 and 5. Non-menstrual pain was reported, respectively, at baseline and 12 month visit by 46 (97.9%) and 15 (38.5%) subjects in the E/P pill group and 49 (89.1%) and 17 (37.0%) of the GNRHa+E/P pill one. The baseline median values of the multidimensional and analog scale were for non-menstrual pain 3 and 5 in the E/P only and 2 and 6 in the GNRHa+E/P group. The corresponding values at the 12 month follow-up visit were 0 and 4 and 0 and 4. These differences between the two groups were not statistically significant. CONCLUSIONS: 1 year after randomization, the two treatment schedules show similar relief of pelvic pain in women with endometriosis.

Immunological parameters in aging: studies on natural immunomodulatory and immunoprotective substances.

Several immune parameters--particularly T-cell dependent immune responses--are altered in aged subjects. To test the hypothesis that they may be the consequence of more general age-related lymphocyte biochemical alterations, and particularly of the energy producing system, the effect of L-carnitine and acetyl-L-carnitine on cell proliferation was studied in peripheral blood lymphocytes from donors of different ages. The results showed that phytohaemagglutinin-induced peripheral blood lymphocyte proliferation was markedly increased in L-carnitine- or acetyl-L-carnitine-preloaded lymphocytes from young and especially from old subjects. Cells from aged subjects considerably improved their defective proliferative capability. Preliminary observations suggest that L-carnitine-preloading also protected peripheral blood lymphocytes from old donors when such cells were exposed to an oxidative stress.

Measurement technique for bottom scattering in shallow water

Sonar performance predictions of reverberation in shallow water rely upon good estimates of the bottom-scattering strength. However, little is understood about bottom scattering in shallow water in the frequency range 400-4000 Hz, particularly its dependency upon frequency and its relationship to the physical properties of the seafloor. In order to address these issues, a new measurement technique has been developed to probe the frequency and angular dependency of bottom-scattering strength. The experimental technique is described which employs either coherent or incoherent sources (lightbulbs). In addition, measurement and modeling results for two diverse shallow water sites are presented. At one site, the scattering appears to arise at or near the water-sediment interface. At the other site, scattering from a 23-m sub-bottom horizon is clearly apparent in the data at and below 1800 Hz. The fact that our measurement technique can directly reveal the presence of sub-bottom scattering is a significant advance in the development of methods to explore the physical mechanisms that control bottom scattering.

[Urinary disorders in the puerperium. Study of the major risk factors]. / I disturbi urinari dell'età puerperale. Studio dei maggiori fattori di rischio.

BACKGROUND: There is a very wide range of genitourinary disorders which can follow vaginal birth, including slight and occasional problems as well as serious disorders which could affect a woman's social and sexual life, for example the effects of dyspareunia on a woman's sexual identity, social marginalization as an inevitable result of symptoms like urinary incontinence, urgency and fecal incontinence. The aim of this study was to identify elements which may be of use in understanding the pathogenetic mechanisms of these disorders. METHODS: Three weeks after birth 537 mothers underwent a clinical genitourinary evaluation including: collection of data regarding pregnancy development and birth, genitourinary history (urinary problem data collected in accordance with the proposal of the International Continence Society), an objective genitourinary examination with a PC-test and identification of possible antagonist abdominal-diaphragmatic muscular synergies, instrumental tests in cases of post-partum urinary incontinence. RESULTS: Maternal age at birth, parity, weight before pregnancy and at term, weight increase, gestational age, duration of the second stage of labour, development and characteristics of birth, perineal condition and neonatal weight were the variables considered as risk factors while genuine stress urinary incontinence, urge incontinence, frequency, urgency, dysuria and inability to interrupt urination were the disorders whose dependence on the various risk factors were to be studied. The analysis of the association of the various pairs of variables recorded both positive and negative correlations, whether the population taken was that of all puerperae or that of only primiparae. CONCLUSIONS: Maternal age at birth, parity and duration of the second stage of labour, even if not always separable from other co-existing risk conditions, are the main responsible risk factors in the pathogenesis of urination disorders in puerperium. These results once again confirm the fundamental role of birth in the pathogenesis of female pelvic statics anomalies and of the genitourinary disorders which are their most evident chemical demonstration.

[Post-partum urogenital and perineal prolapse]. / Il descensus urogenitale e perineale nel postpartum.

BACKGROUND: It is well known that vaginal birth, even under apparently normal circumstances, involves a significant mechanical straining of the various muscular connective structures which make up the pelvic floor and that an unusual strain of the perineal plates can cause morphologic-functional alterations which are not entirely reversible. The integrity of structures which make up the "pelvic floor" and the "endopelvic fascia" is the fundamental element to maintaining a normal anatomic position of the pelvic organs in the various functional conditions. Consequently prolapse of female pelvic organs can be linked back to the functional limitations of perineal plates (muscular support fascia system) and/or of the ligaments of the sub-peritoneal endopelvic connective tissue (ligament suspension system). METHODS: After birth 537 mothers underwent a urological and gynecological examinations as follows: collection of clinical data regarding pregnancy development and birth; medical history regarding the number of day- and night-time urinations, urinary volume, possible encouraging factors and pre-urinary sensations; objective urological and gynecological examination (pubo-coccygeal test, highlighting of possible agonistic and antagonistic muscular synergies, stress test, evaluation and staging of vaginal prolapse according to Baden and Walker; instrumental evaluation in cases of post-partum urinary incontinence. Simple regression analyses were carried out where prolapse of various vaginal segments were proportionately related to the various risk factors. RESULTS: Maternal age at birth, parity, weight before pregnancy and at term, weight increase, duration of second stage of labour, development and characteristics of the birth, perineal condition and neonatal weight were all variables considered risk factors while prolapse in each vaginal segment, PC-test, involuntary reflex execution of opposite command and uterine retroversion were all "response variables" whose dependence on various risk factors was studied. Analysis of the associations between the various pairs of variables showed a correlation, both positive and negative, whether the population considered was that of all mothers or that of primiparae. CONCLUSIONS: In the light of the results of this study, it can be said that there are two important pathogenetic factors: the tissue factors and the iatrogenic factor. Elevated maternal age and multiparity underline the role of the tissue factor in the pathogenesis of obstetric perineal damage. With regard to the iatrogenic factor it is interesting to note a higher concentration of symptomatic women cases where labour had been induced or birth had been achieved through instrumental delivery.

[Anthropometric skin-fold and impedance measurement assessment of the maternal nutritional status at the end of pregnancy. Comments deduced from a study of 54 cases]. / Valutazione antropoplicometrica ed impedenziometrica dello stato nutrizionale materno al termine della gravidanza. Considerazioni dedotte dallo studio di 54 casi.

BACKGROUND: It is generally accepted that an optimal maternal weight gain is indicative of a normal course of pregnancy. Nevertheless, evaluations of nutritional status should not be limited to the periodic body weight measurement alone, they should rather include the measurement of anthropoplicometric parameters more strictly associated with the variations of the different body components (fat mass, fat-free mass and body water). Therefore, it seems necessary to define standard parameters, such as to allow the attribution of a clinical value to the anthropometric variations that can be measured during pregnancy. In this study we preliminarily proposed to survey maternal nutritional status in a group of women examined in the second day after delivery. MATERIALS AND METHODS: Anthropoplicometric parameters and bioelectric impedance measurements were evaluated in order to study nutritional status of 54 puerperae within 48 hours after delivery. A linear regression analysis was carried out examining the relationship of anthropoplicometric measurements with gestational weight gain and post-partum impedance parameters. RESULTS AND CONCLUSIONS: The results of this study lead to believe that a correct examination of maternal nutritional status must necessarily include not only weight and body mass index, but also an accurate and periodic anthropoplicometric evaluation including limb circumferences and triceps plica.

[The role of pelvic outlet dystocia in the pathogenesis of pelvic floor damage cause by obstetric intervention]. / Ruolo delle distocie dell'egresso pelvico nella patogenesi del danno perineale ostetrico.

BACKGROUND: Vaginal delivery may cause injury to the perineum even when it progresses normally. In the light of this, some authors have suggested that other factors besides obstetric trauma may be implicated in the pathogenesis of genito-urinary prolapse, such as maternal constitution, race, connective tissue characteristics, etc. The aim of this study was to investigate the role of mechanical factors in causing perineal injury. METHODS: We examined the clinical data of 250 women who underwent, 20 days after delivery, a clinical examination consisting of a semeiological study of micturition and a functional evaluation of the pelvic floor. RESULTS: Outlet dystocia seems to be associated with the higher prevalence of anteriormedian prolapse and seem to play an important role in the pathogenesis of post-partum pelvic floor deficiency. CONCLUSIONS: If valid prognostic indexes for the selection of patients at risk are found, it will be possible to carry out an effective prevention of pelvic floor damage and also to avoid an indiscriminante and unjustified increase of laparotomic deliveries.

[Prevention of obstetrical damage to the perineum. Proposal for a fetal-pelvic index for the selection of parturients at risk]. / Prevenzione del danno perineale ostetrico. Proposta di un indice feto-pelvico per la selezione delle partorienti a rischio.

BACKGROUND: According to some authors, vaginal delivery always causes denervation of perineum and the greater the damage the longer the second labour phase (the so-called "delivering phase"). Therefore, it is necessary to reduce the number of too prolonged labours, but it is equally important to avoid an uncontrolled increase of cesarean sections. In order to achieve this objective, it is important to carry out a careful selection among laboring women and choose those most at risk for whom cesarean section is strongly recommended. On the basis of the data collected by the medical literature and in consideration of the pathogenetic role of the outlet dystocia, we have tried to identify a simple and effective prognostic index resulting from the different pelvimetric and ultrasonographic parameters. METHODS: In 72 full-term pregnant women, we have taken into account the ultrasonographic parameters expressing the fetal dimension (cephalic diameters, cephalic and abdominal circumferences, estimated fetal weight according to Haddlok), the outlet pelvic diameters (trans-ischial and coccygeal-pubic) and a fetal-pelvic index derived from these parameters. RESULTS: If taken individually, these parameters do not seem to have any direct connection with the length of the delivering phase, but the combination of the cephalic and external pelvimetric diameters has produced a significative statistical coefficient. CONCLUSIONS: On the basis of the data collected, it is suggested that a careful evaluation of external pelvimetric and cephalic parameters would be useful from the clinical point of view.

[An inguinal hernia with a strangulated Meckel's diverticulum (Littré's hernia). A case report and review of the literature]. / Ernia inguinale con diverticolo di Meckel strozzato (ernia del Littré). Osservazione di un caso e revisione della letteratura.

The A. describe a case of Littré's hernia drawn from personal experience. The article discusses Meckel's diverticulum complications, difficult presurgical diagnosis and different surgical techniques of performing diverticulectomy.

[The surgical treatment of gastric cancer. A review of the results and a comparison with Japanese experience]. / Il trattamento chirurgico del cancro gastrico. Revisione dei risultati e confronto con l'esperienza giapponese.

The authors synthetize the principal trends about surgical treatment of gastric cancer underlining the actual way. Then, is considered the Japanese experience whose results are compared with those in the west to try an explanation of surgical data amply in favour of resected in Japan.

[Hypocalcemia after total thyroidectomy. Therapeutic considerations]. / La ipocalcemia post-tiroidectomia totale. Considerazioni terapeutiche.

One of the post-operative complications of total thyroidectomy is the onset of a transitory or permanent hypocalcemic syndrome. Hypocalcemia is caused by different physiopathological mechanisms that operate either individually or in association (parathyroidal deficit, deficiency of bone calcium subsequent to parathyroid osteodystrophy, acute emission of calcitonin into the circulation). Calcemia levels were studied p.o. in a selected group of 25 patients who had undergone total extracapsular thyroidectomy with the apparent preservation of the parathyroid glands. Totals thyroidectomy has been performed in 15 patients with euthyroid goitre, in 4 with papillary carcinoma, in 3 with diffuse toxic goitre and in 3 with multinodular toxic goitre. Eight patients received drug and diet therapy since calcemia levels were lower than 7.5 mg/ml. Fifteen days after the operation, calcemia levels in 21 patients were subliminal and dihydrotachysterol therapy was commenced. Hypocalcemia was permanent in only one case.