Induction of mitochondrial dysfunction in patients under cardiopulmonary by-pass: preliminary results.

OBJECTIVE: Cardiac surgery is often performed by cardiopulmonary by-pass (CPB), generally associated with organ dysfunction. The aim of this work was to determine if and how this phenomenon is related to mitochondrial damage. To this purpose, the effect of the addition of serum from CPB patients to human fibroblasts cultures on mitochondrial respiratory chain and oxidative phosphorylation (OXPHOS) activities was investigated. PATIENTS AND METHODS: Serum samples of five patients were obtained before (pre-CPB) and after 6 h CPB weaning (CPB). Mitochondrial OXPHOS activities were examined by polarographic and spectrophotometric assays, and reactive oxygen species (ROS) production was measured by a spectrofluorimeter. RESULTS: Addition of CPB serum to fibroblasts determined a decrease of mitochondrial oxygen consumption due to an inhibition of mitochondrial respiratory chain and some OXPHOS enzymes activities. This inhibition seems to be mainly related to a reduced activity of complex I. CONCLUSIONS: Our data represent the first translational research evidence showing that CPB determines mitochondrial dysfunction which leads to impairment of OXPHOX activities and to an increase in ROS production, compromising tissue bioenergetic efficiency.

Effect of low-level laser irradiation on osteoblast proliferation and bone formation.

Applications of laser therapy in biostimulation and healing injured tissues are widely described in medical literature. The present study focuses on the effects of laser irradiation on the growth rate and differentiation of human osteoblast-like cells seeded on titanium or zirconia surfaces. Cells were laser irradiated with low therapeutical doses at different intervals and the effects of irradiation were evaluated at each time-point. After 3 hours lasered cells showed an enhanced mitogen activity compared to non-lasered control cells and a higher alkaline phosphatase activity, marker of bone formation. At the same time, the mRNA of RUNX2 and OSTERIX, two genes involved in osteoblast differentiation, showed a clear decrease in lasered cells. This reached the lowest value 6 to 12 hours after irradiation, after which the transcripts started to increase, indicating that the laser treatment did promote the osteogenic potential of growth-induced cells. These results indicate that Low Level Laser Treatment (LLLT) stimulates osteogenic cell proliferation.

Differential effects of all-trans retinoic acid on the growth of human keratinocytes and mouth carcinoma epidermoid cultures. Involvement of GRIM-19 and complex I of the respiratory chain.

Squamous cell carcinoma (SSC) is the most frequent malignant tumor of the oral cavity. A study on the effect of all-trans retinoic acid (RA) on cell growth, expression of GRIM-19 and content and activity of complex I of the mitochondrial respiratory chain in normal human keratinocytes (NHEK) and mouth carcinoma cells with low (HN) and high (KB) transformation grade was carried out. In NHEK cells, RA treatment resulted in growth suppression, significant overexpression of GRIM-19 protein, enhanced content of complex I but depressed activity of NADH-UQ oxidoreductase activity of the complex. In HN cells, RA treatment depressed cell growth, inhibited the enzymatic activity of complex I but had no significant effect on the levels of GRIM-19 and complex I. In KB cells RA had no effect on cell growth, GRIM-19 expression, content and activity of complex I.

[Ovulation induction using pulsatile GnRH in patients with hypothalamic amenorrhea: effects of changing the frequency and dosage on the luteal phase]. / Induzione dell'ovulazione con GnRH pulsatile in soggetti affetti da amenorrea ipotalamica: effetti della modificazione della frequenza e del dosagio sulla fase luteale.

Effects of frequency and dose variation of GnRH administration by minipumps Zyklomat, were studied in five women during luteal phase. As controls were assumed the same patients; in fact they had been previously treated with pulsatile GnRH with constant dose and frequency all over the cycle. Then the patients received the same treatment but the dose and the frequency of infusion were changed in the luteal phase. Our data show an increase of mean levels of progesterone and oestradiol during the luteal phase and a better pregnancy rate when the frequency and the dose of GnRh administration were changed.

[Flare-up study and gonadotropin extinction curve in patients treated with long-acting triptorelin, 3.75 mg im]. / Studio del Flare-Up e delle curve di estinzione delle gonadotropine in pazienti trattate con Triptorelina Long Acting 3,75 mg I.M.

Flare-Up study and gonadotropin-oestradiol extinction pattern were examined in two groups of 11 and 10 patients during treatment with Triptorelin LA. Plasmatic gonadotropin and oestradiol levels were evaluated by LIA and RIA respectively. The resulting data confirm that LH levels show an increase sixfold than basic values (Flare-Up effect), while FSH levels increase twice after two days of treatment. This effect was exhausted in five days. Gonadotropin-oestradiol extinction pattern demonstrates a condition of hypogonadotropic hypogonadism after two weeks of treatment with LHRHa. This study could explain some aspects of a LHRH-a action, namely for their clinical use in ovulation induction protocols.

[Estradiol benzoate test in the study of pituitary block induced by triptorelin]. / Studio dinamico del blocco ipofisario indotto dalla triptorelina, mediante test all'estradiolo benzoato.

The suppression of the positive oestradiol (E2) feed-back mechanism was studied in 21 patients during treatment with long acting GnRHa-triptorelin 3.75 mg (Ipsen-Biotech). A basal sample for FSH, LH and E2 evaluation was taken before a single administration of 2.5 mg of oestradiol benzoate (EB) i.m. at 24 and 48 hours later. The test was performed 1, 2, 3, 4, 5, 6 and 7 weeks after i.m. injection of GnRHa. The E2 concentration 24 hours after EB injection was > 400 pg/ml, able to induce, in physiological conditions, a positive feedback. In the other hand no FSH an LH rise was observed in all test after 1, 2, 3, 4, 5, 6 and 7 weeks from GnRHa administration. Our results show the absolute suppression of the positive feed-back mechanism during the treatment with a single dose of long acting tryptorelin 3.75 mg i.m., the same results were obtained until the 7th week after GnRHa administration.

Endocrine abortion in assisted reproduction technologies (ART).

Luteal function, endometrial receptivity, endometrial prolactin and glycoprotein secretions, blastocyst-secreted immunomodulant factors and embryo quality are nowadays considered the main determinants involved in embryo implantation control. The endometrial factors are progesterone-dependent. Out of 128 cycles of ART (AIH-IU, GIFT, IVF-ET), performed in 67 women at the Dept. of Obstetrics and Gynaecology of Parma during the period 1986-1991, 31 conceptions were obtained (pregnancy rate: 24.21%), 7 of which miscarried (abortion rate: 22.58%). According to these data, ART high abortion rate is possibly connected with poor luteo-endometrial function and poor embryo quality. Controlled ovarian hyperstimulation is thought to inhibit embryo implantation after IVF-ET by decreasing endometrial receptivity. Such a situation can be treated either by exogenous progesterone administration or by tubal techniques (GIFT, TET) performed in non-tubal infertility. Both strategies showed to better endometrial receptivity. A delayed intrauterine embryo transfer at blastocyst stage, when cocultures allow to, is supposed to raise the implantation rate in tubal infertility by enhancing embryo selection and endometrial receptivity. This paper also reports preliminarily on the predictive value of beta-HGC and estradiol levels, as well as of endometrial thickness, on early pregnancy outcome.

Gallstones and uterine fibroids.

In a series published in 1961, an unusual frequency of hysterectomies for uterine leiomyomas (fibroids) was reported in women with gallstones. The purpose of this study was to confirm the association between gallstones and uterine leiomyomas with a patient control study and to investigate its physiopathologic basis comparing the cholesterol saturation of bile in women with gallstones, in women with leiomyomas but no gallstones and in those in the control group with no gallstones or leiomyomas. Patients admitted to the surgical department have, routinely, echography of the gallbladder before and manual exploration of the pelvic floor during surgical intervention. For the first part of the study, we collected information concerning the diagnosis of leiomyomas from the operating room registers and about the diagnosis of gallstones from the clinical records. In 1982, 42 of 139 women operated upon consecutively for gallstones and five of 69 operated upon for other diseases of the gastrointestinal tract had leiomyomas, a statistically significant difference (chi-square test, p less than 0.001). This difference persisted stratifying women with gallstones and those in the control group for age. In the second part of the study, we examined the bile collected at duodenal drainage after gallbladder stimulation with cholecystokinin, in 11 young women with radiolucent gallstones (echography and cholecystography), in ten women with leiomyomas (gynecologic examination and pelvic echography) but no gallstones (echography) and in 11 women with no leiomyomas (gynecologic examination or pelvic echography) or gallstones (echography). Cholesterol, phospholipids and total bile acids in the biliary tract were analyzed with standardized enzymatic methods. The cholesterol saturation index of the biliary tract was higher in patients with leiomyomas than in those in the control group (Wilcoxon rank sum test, p less than 0.01) and similar to that of women with radiolucent gallstones. These data suggest that gallstones and leiomyomas are associated diseases, probably sharing a common cause.

Preliminary observation on human tubal electrical activity in vivo.

The authors have set up a new technique for recording the human tubal electrical activity in vivo. A polyethylene catheter was used with six couples of electrodes placed on the tube of 5 women during simple laparohysterectomy. The slow electrical activity was recorded by means of a 8-channel paper amplifying recorder for 3-5 days from surgery. The fimbria showed frequencies between 1.25 and 3.3 c/min, the ampulla showed higher frequencies (1-5 c/min), the isthmus showed little electrical activity. The propagation signal velocity ranges between 0.25 and 0.5 cm/sec. The amplitude ranges between 50 and 200 V. The propagation occurs mainly in the distal-proximal direction.

Estetrol and utero-placental flow after progesterone load.

On the basis of recent demonstration in animals of the effect of some hormones on uteroplacental flow, the Authors examined the response of plasmatic Estetrol (15 alpha-hydroxy-estriol) after the administration of progesterone to pregnant women with low Estrogen values. The increase of this compound was related to an improvement of placental function, probably dependent on an increase of available O2, and therefore on uterine blood flow. This can justify a progesterone treatment in such pregnancies.