Beneficial effects of melatonin on nicotine-induced vasculopathy.

Tobacco smoking is responsible for death of many people each year and increases the risk of developing numerous disorders, particularly cardiovascular disease and cancer. Among the components of cigarette smoke, nicotine is known to excert proatherosclerotic, prothrombotic and proangiogenic effects on vascular endothelial cells. The current study was designed to investigate the mechanisms by which nicotine induces endothelial dysfunction and further to examine whether melatonin protects against nicotine-induced vasculopathy. Four groups of male rats (controls, melatonin-treated, nicotine treated [100 microg/mL in drinking water], and nicotine plus melatonin [5 mg/kg/day] treated) were used in this study. After 28 days all the animals were killed by decapitation and the aorta was removed. We evaluated the hydroxyproline content, and the different expression of proteins involved in several types of stress (ERK1/2), in fibrosis (TGF-beta1, NF-kappaB) and in recruitment of circulating leukocytes onto the vessel wall, including intercellular adhesion molecule-1 (ICAM-1) and vascular cellular adhesion molecule-1 (VCAM-1). These metabolic pathways are important in the development of nicotine-induced atherosclerosis and hypertension. Our results show that nicotine induces marked structural and functional alterations in the aorta. Nicotine receptor binding results in activation and phosphorylation of ERK 1/2. This enzyme, in turn, activates both TGF-beta1 and NF-kappaB; they stimulate respectively the synthesis of type I collagen, responsible of fibrosis, and moreover ICAM-1, VCAM-1 and reactive oxygen species. Based on these findings, melatonin is able to minimize the negative effects of nicotine by blocking the activation of ERK and the other signalling pathways in which this enzyme is involved.

Red wine polyphenols prevent cyclosporine-induced nephrotoxicity at the level of the intrinsic apoptotic pathway.

Flavonoids, polyphenol derivatives of plant origin, possess a broad range of pharmacological properties. A number of studies have found both pro/anti-apoptotic effects for many of these compounds. For these reasons we investigated whether Provinols flavonoids obtained from red wine, have anti-apoptotic properties. The investigations have been carried out in rats treated with Cyclosporine A (CsA). In particular, four groups of rats have been treated for 21 days with either olive oil (control group), with CsA, with Provinols, or with CsA and Provinols simultaneously. Oxidative stress, systolic blood pressure, body weight, biochemical parameters and different markers of pro/anti-apoptotic pathway were measured. CsA produced an increase of systolic blood pressure, a decrease in body weight, serum creatinine levels, urinary total protein concentration and creatinine clearance. Moreover, CsA induced renal alterations and the translocation of Bax and cytochrome c from cytoplasm to mitochondria and vice versa. These changes activated the caspase cascade pathway, that leads to morphological and biochemical features of apoptosis. Provinols restored morphological and biochemical alterations and prevented nephrotoxicity. In conclusion, this study may augment our current understanding of the controversial pro-/anti-apoptotic properties of flavonoids and their molecular mechanisms.

Evaluación de alteraciones hemorreológicas en pacientes hipertensos / Hemorheological alterations in hypertensive patients

El objetivo de este trabajo fue investigar el perfil de viscosidad sanguínea y evaluar la influencia de factores plasmáticos ( fibrinógeno) y celulares ( agregación eritrocitaria ) en un grupo de pacientes hipertensos comparados con un grupo de paciente normotensos. Se trabajó con sangre anticoagulada de pacientes hipertensos no diabéticos (n=3) e indivíduos sanos (n=40). La viscosidad plasmática y de sangre entera se determinaron con un viscosímetro cono-plato. La agregación eritrocitaria se estudió por observación microscopia de los agregados y cuantificación a través de un parâmetro de forma denominado ASP ( Aggregation Shape Parameter), definido como la relación de área proyectada respecto al perímetro. El fibrinógeno se determino con un coagulómetro por el método de Clauss. Los valores de viscosidad de sangre entera resultaron significativamente aumentados en los pacientes hipertensos respecto de los normales para todas las velicidades estudiadas. Los valores de viscosidad plasmática solo presentaron diferencia significativas a bajas velocidades de corte (1.15 a 11.56 seg •1) . Los pacientes hipertensos presentaron agregados amorfos e irregulares, lo que se refleja en los valores alterados de ASP, significativamente mayores (p<0.001) en paciente hipertensos (0.69± 0.11) respecto de los indivíduos normales ( 0.25± 0.12). Los valores de fibrinógeno resultaron ligeramente superiores en los pacientes hipertensos respecto del grupo control (p< 0.01). Numerosos parámetros hemorreológicos juegan un papel importante en la patogénesis de la hipertensión. Entre estos factores hemorreológicos, valores parâmetros podrían estar en la hipertensión ( hematrocito, fibrinógeno plasmático, deformabilidad y agragación eritrocitaria , viscosidad sanguínea y plasmática). En este trabajo, se pudo demostrar anormalidades en la agregación eritrocitaria, detectada por los valores de ASP que podría estar involucrado en las complicaciones vasculares de la hipertensión.

Modulation of pro- and anti-apoptotic genes in lymphocytes exposed to bone cements.

The ability of bone cements to modify the apoptotic program in activated immune cells and the mechanisms by which they act were evaluated. Mononuclear cells were collected from healthy individuals, cultured for 4 and 24 h with phytohemoagglutinina-P and cement extracts and then tested to assess: (a) cell viability; (b) early apoptotic events, by Annexin V/propidium iodide staining; and (c) the expression of pro- (p53, c-myc, ICE) and anti-apoptotic (bcl-2) genes. After 4 h three cements were able to increase significantly the percentage of apoptotic cells, while after 24 h no differences were found. The proportion of dead cells was not significantly changed at either culture time. The simultaneous expression of both pro-apoptotic (ICE, c-myc, p53) and antiapoptotic genes (bcl-2) was investigated only with regard to the materials which induced significant changes in apoptosis: two cements induced the p53 expression, while the third down-regulated bcl-2. As apoptosis regulates the balance of immune response, the authors recommend that the interaction between materials and immune cells should be assessed, so that the use of pro-apoptotic materials may be avoided in patients with immune defects.

The long styloid process syndrome or Eagle's syndrome.

Eleven cases of Eagle's syndrome or long styloid process syndrome are presented. It is a rare entity, which is not commonly suspected in clinical practice. Symptoms were dull and persistent pharyngeal pain, dysphagia, and facial pain. In addition to careful clinical examination, the use of plain radiographs, orthopantomograms or CT scan has allowed accurate preoperative diagnosis of this syndrome. Surgical treatment has positively resolved the symptoms in these cases. The International literature is discussed regarding anatomy, symptoms and treatment of the long styloid process syndrome.

In vitro cytokine production by mononuclear cells exposed to bone cement extracts.

The authors evaluated the ability of bone cement to modify the profile of pro-inflammatory cytokines secreted by the immune cells. Peripheral blood mononuclear cells (PBMC) collected from healthy individuals were cultured with cement extracts and tested to assess the release of IL-1beta, TNFalpha, GM-CSF and IL-6 in both unstimulated and PHA-stimulated PBMC. The cytokine release of unstimulated PBMC was very poor, and in particular the IL-1beta was undetectable: the addition of cement extract increased both TNFalpha and GM-CSF release and decreased IL-6, sometimes significantly. The most recurrent observation in PHA-stimulated PBMCs exposed to bone cement extract was the increase in both IL-1beta and IL-6 release, while both the mean concentration and the index of release of TNFalpha and GM-CSF were changeable. In conclusion our results showed that leachable components of some bone cements can induce in vitro the release of pro-inflammatory cytokines which are known to be involved in the bone resorption associated with aseptic loosening of hip prostheses. These findings allowed us to identify materials endowed with the highest inflammatory power.

Identification and characterization of an 18-kilodalton, VAMP-like protein in suspension-cultured carrot cells.

Polyclonal antibodies raised against rat vesicle associated membrane protein-2 (VAMP-2) recognized, in carrot (Daucus carota) microsomes, two major polypeptides of 18 and 30 kD, respectively. A biochemical separation of intracellular membranes by a sucrose density gradient co-localized the two polypeptides as resident in light, dense microsomes, corresponding to the endoplasmic reticulum-enriched fractions. Purification of coated vesicles allowed us to distinguish the subcellular location of the 18-kD polypeptide from that of 30 kD. The 18-kD polypeptide is present in the non-clathrin-coated vesicle peak. Like other VAMPs, the carrot 18-kD polypeptide is proteolyzed by tetanus toxin after separation of coatomers. Amino acid sequence analysis of peptides obtained by digestion of the 18-kD carrot polypeptide with the endoproteinase Asp-N confirms it to be a member of the VAMP family, as is suggested by its molecular weight, vesicular localization, and toxin-induced cleavage.

Cytokine release in mononuclear cells of patients with Co-Cr hip prosthesis.

The aim of this study was the evaluation of the release of bone-resorbing cytokines in peripheral blood mononuclear cells (PBMC) of patients with aseptic loosening of Co-Cr hip prostheses. TNF-alpha, IL-6, and GM-CSF were measured in both unstimulated and PHA-stimulated PBMC, and in PBMC cultured in the presence of chromium and cobalt extracts. Blood samples from healthy donors were used as controls. Serum samples of both patients and healthy donors were tested to determine the concentration of metal ions. The proportion of lymphocyte, monocyte and lympocyte subpopulation in cultured PBMC did not differ in patients and the control group. In unstimulated PBMC the release of TNF was significantly higher in patients than in the control group, while IL-6 was significantly decreased and no change was observed for GM-CSF. When the PBMC were challenged with chromium extract, all the 'index of cytokine release' resulted higher in patients than in the control group; cobalt extract increased both the TNF and GM-CSF index, but not the index of IL-6 release. Metal concentrations in serum from patients were significantly increased and correlated with the TNF release in PBMC stimulated with both metal extract. Our results suggest that a CoCr-implant releases a large amount of metal ions which could mediate the priming or the renewal of a cell-mediated hypersensitivity reaction. The prevalence of circulating lymphocytes responsible for the delayed hypersensitivity, namely Thl, would justify both the significant increase of TNF and the significant decrease of IL6 in unstimulated PBMC of patients, as well as the significant increase of the 'index of cytokine release' after the challenge with metal ions.

[Laparoscopy-assisted hysterectomy and laparoscopic preparation. Apropos of a series of 177 cases]. / L'hystérectomie coelio-assistée et coelio-préparée. A propos d'une série de 177 cas.

Our objective was to determine the limits of laparoscopic-assisted vaginal hysterectomy (LAVH) and the value of a preoperative scoring system to determine the operative approach to hysterectomy. Between January 1991 and December 1996, 152 out of 177 patients had LAVH and 25 had laparoconversion. The mean operating time was 163 min. The overall postoperative complication rate was 8.4%. The hospital stay was 4.8 days for LAVH versus 6.2 days for laparoconversion (p < 0.01). For each patient, a preoperative scoring system was established according to uterine size, previous laparotomy, uterine mobility, pelvic adhesions and endometriosis stage. The laparoconversion rate increased according to the score, as it was 7.8% for a score < or = 7 and 80% for a score > 7. LAVH offers a technique to convert some abdominal hysterectomies into vaginal hysterectomies. The use of the preoperative scoring system may help to determine patients who may benefit from the laparoscopic route and those with a high risk of laparoconversion.

Hipertensión arterial y marcadores de grupo sanguíneo: asociación con el polimorfismo MN / Hypertension and blood group markers: association with MN polymophins

El objetivo de este trabajo fue analizar la relación entre presión arterial y marcadores genéticos eritrocitarios MN. La expresión de los antígenos M y N, se investigó por el Test de hemaglutinación en tubo, en una muestra de 164 individuos clasificados según presión arterial (normales e hipertensos) y fenotipo MN. De los 53 pacientes hipertensos, los porcentajes de cada fenotipo fueron: MN 56,6 por ciento,MM 34,0 por ciento, NN 9,4 por ciento mientras que entre los 111 individuos normotensos, la distribución resultó: MN 70,3 por ciento, MM 18,9 por ciento y NN 10,8 por ciento. De los resultados obtenidos en este trabajo, por la Prueba x², se concluye que sólo existe asociación entre el fenotipo MM y la hipertensión arterial. Diferencias en el patrón de agregación, debido a diferencias estructurales de los distintos grupos sanguíneos, podrían generar susceptibilidad a cambios en la viscosidad sanguínea, afectando el proceso de perfusión. El estudio de marcadores eritrocitarios, que pueden ser determinados por metodologías accesibles, podrían aportar datos de interés para la investigación de factores de riesgo en el desarrollo de la hipertensión arterial

The secretory nature of the lesion of carrot cell variant ts11, rescuable by endochitinase.

The carrot cell variant ts11 is unable to form somatic embryos at the non-permissive temperature of 32 degrees C, but the block can be overcome by the addition of a 32-kDa acidic endochitinase to the medium. In this work we conducted a cyto-histological analysis of the blocked embryo forms. The morphology of the endomembrane system is altered; in particular, the ER is dilated and may show electron-dense precipitates and continuity with the plasma membrane. These morphological alterations do not occur in the presence of externally-added endochitinase. We also noticed modifications of the culture medium that are probably related to the morphological observations: the total amount of secreted proteins is reduced and pulse-chase experiments revealed that, compared with wild-type cells, the secretion of major polypeptides is reduced while new minor polypeptides are secreted. Western blot analysis revealed the presence of the binding protein BiP, a resident of the ER and of glutamine synthase, a cytosolic protein, in the medium of ts11 but not wild-type cells. These results indicate that ts11 is altered in the secretory pathway but do not clarify the role of endochitinase.

Results of long-term experimental studies on the carcinogenicity of methyl tert-butyl ether.

Methyl-tert-butyl ether (MTBE) was submitted to long-term carcinogenicity bioassays on Sprague-Dawley rats. The test compound was delivered in olive oil by stomach tube (gavage), at the doses of 1000, 250, and 0 mg/kg b.w. to groups of 60 males and 60 females, once daily, 4 times weekly, for 104 weeks. All animals were kept under control until spontaneous death. MTBE was found to cause in males an increased incidence of Leydig cell testicular tumors in the group treated with the higher dose, and in females a dose-related increase of leukemias, an increase of dysplastic proliferations of lymphoreticular tissues, and also an increase of uterine sarcomas at the lower tested dose. On the basis of the presented data, MTBE must be considered a potential carcinogen.

Transvaginal intratubal methotrexate treatment of ectopic pregnancy. Report of 100 cases.

Between November 1988 and December 1993, 100 patients with a common, unruptured ectopic pregnancy were treated with 1 mg/kg injection of intratubal methotrexate under transvaginal sonographic control. Patients were not excluded from this series on the basis of the size of the adnexal mass, the term of ectopic pregnancy or initial beta-human chorionic gonadotrophin (HCG) concentrations. Patients were excluded following uncertain diagnosis, signs of a ruptured ectopic pregnancy, or a significant haemoperitoneum on ultrasound scans. The mean age of the patients was 29.5 years (range 20-41). The mean gestational age and initial HCG concentration were 7.5 weeks (5-11) and 11,614 mIU/ml (192-105,000 respectively). Of the 100 patients, 22 (22%) had an ectopic pregnancy with active cardiac activity. Complete resolution was obtained in 78 out of these 100 ectopic pregnancies. Of these, 66 patients (85%) needed only one intratubal methotrexate injection, and 12 patients (15%) required a second i.m. methotrexate injection of 1 mg/kg. In this study, local treatment with one single intratubal methotrexate injection was successful in only 66% of patients. The mean resolution time for reduction of beta-HCG concentrations was 23.5 days (range 7-40). There was no statistically significant correlation between initial beta-HCG concentrations and outcomes after methotrexate treatment of ectopic pregnancy in our study. Where embryonal heart beats were observed, the success rate of the procedure was 40.9% (nine out of 22 cases). In the absence of cardiac activity, or when ultrasound examination showed no embryo, the success rate achieved was 84.6% (66 out of 78 cases) (P < 0.01). In all, 34 patients were considered to be incompletely cured after only one intratubal methotrexate injection: 12 patients required a second i.m. injection, a stagnation of beta-HCG concentrations was observed in 15 patients, abdominal pain occurred in six patients, and one patient suffered tubal rupture with haemoperitoneum. A total of 22 patients required secondary surgical management (salpingectomy). No biochemical or clinical side-effects of methotrexate treatment occurred. Tubal alteration ascribable to methotrexate injection occurred in one patient in our study. Out of 75 patients in this series who wished to conceive, 21 (28%) became pregnant within 1 year with the following outcomes: 11 pregnancies at term, three miscarriages, one induced abortion and six recurrent ectopic pregnancies (four occurred on the same side). Our findings suggest that treatment of common unruptured ectopic pregnancy without prior selection of patients, by a single intratubal methotrexate administration was associated with a 66% success rate. This was dependent only on the presence of embryonal heart beats and there was no correlation between the success rate and initial beta-HCG concentrations. Successful outcome after methotrexate administration for ectopic pregnancy could be perfected by way of an improved selection of patients based on inactive embryonal hearts and absence of a visualized embryo.

[Distal tubal surgery: a critical review of 104 laparoscopic distal tuboplasties]. / Chirurgie de la trompe distale. Analyse critique d'une série de 104 plasties tubaires distales coeliochirurgicales.

MATERIALS AND METHODS: Between January 1988 and June 1993, laparoscopic distal tuboplasty was carried out on 104 patients. The mean age of the patients was 32 years: the duration of infertility was 3.2 years; infertility was primary for 44 patients (42.3%) and secondary for 60 patients (57.7%). The patients were classified according to the French cooperative tubal scoring system: stage 1: 21 patients, stage II: 49 patients, stage III: 22 patients, stage IV: 12 patients; 37 patients underwent a fimbrioplasty (35%) whereas 67 patients underwent a neosalpingostomy (65%). No complications were observed either during or after the operation. All patients were discharged two or three days after the operative procedure. Antibiotic and antiinflammatory treatments were systematically administered post-operatively. RESULTS: Thirty-four patients obtained intrauterine pregnancies (32.5%) and 5 patients obtained ectopic pregnancies (4.8%); 76% of the intrauterine pregnancies were obtained within the 1st postoperative year. The pregnancy outcome was directly correlated to the tubal scoring system. Intrauterine pregnancy rate decreased with the severity of the tubal damage: stage 1: 12/21 (57%), stage II: 19/49 (38.7%), stage III: 3/22 (13.6%), stage IV: 0/12. CONCLUSION: Laparoscopic surgery represents the first-choice treatment of distal tubal occlusion in stage I and II. In stage IV, IVF should be suggested immediately. In stage III, the choice of treatment is more difficult: the main prognostic factor might be the tubal mucosal appearance.

[Laparoscopy-assisted and -prepared hysterectomy. A series of 80 cases]. / Hystérectomie coelio-assistée et coelio-préparée. A propos d'une série de 80 cas.

OBJECTIVE: Our objective was to determine the interest of laparoscopic assisted vaginal hysterectomy. STUDY DESIGN: Between January 1991 to december 1994, 80 patients had laparoscopically assisted vaginal hysterectomy. We reviewed with particular emphasis characteristic indications, complications. RESULTS: Eighty were performed as laparoscopically assisted vaginal hysterectomy. 14 patients (17.5%) had laparotomy conversion; because of size of uterus in 3 cases, suspected ovarian tumor in 3 cases. Pelvic adherences in 4 cases, urinary tract injuries in 1 case, hypercapnia in 1 case, hemorrhage in 2 cases. 9 patients experienced febrile morbidity and 1 urinary infection. 1 patient received 2 units of packed red blood cells. The hospital stay was 5 days for laparoscopically assisted vaginal hysterectomy versus 5.9 for laparotomic hysterectomy. CONCLUSION: Laparoscopically assisted vaginal hysterectomy offers a technique to convert certain abdominal hysterectomies into vaginal hysterectomies with a 17.5% laparoconversion rate.

[Comments on organization and activity of ENT day hospital department: three years of experience]. / Riflessioni su ...organizzazione ed attività di un reparto di day hospital in O.R.L.: esperienza di un triennio.

The authors deal with organizational and technical aspects concerning otorhinolaryngological day hospital's management. They explain purposes and reasons leading to the establishment of this sort of service; they analyse related problems and the adopted methodologies for simplifying admission procedures. They report a statistical survey regarding the period from May 1991 to May 1994, concerning surgical and medical operations, and instrumental diagnostic researches performed at the IV ENT Department of "La Sapienza" University of Rome. The statistical analysis concerning the 1526 patients sent to this department, shows the downright prevalence of surgical operations performed in operating room (52%). Ambulatory operations and instrumental diagnostic researches account respectively for 39.6% and for 29.3% of all hospitalizations. Among all surgery operations performed in operating room stand out the notice incidence of therapeutical and diagnostic microlaryngoscopies. The most frequent ambulatory operations were the removal of tracheal cannula (21%), while TC was the most frequent of all instrumental diagnostic researches. A careful selection of patients, on the basis of their general condiction, account for the low incidence of subjects not discharged within 24 hours. After having analyzed the survey's data, the authors underline advantages and finalities of day hospital: a shorter hospital stay, reduction of hospitalization's costs, and a larger related number of patients to be contented. Finally they hope that this sort of service will spread more.

[Ectopic pregnancy: criteria to choose therapy]. / Grossesse extra-utérine: critères du choix thérapeutique.

The ectopic pregnancy (EP) could be treated by spontaneous resolution, or medical treatment or laparoscopic treatment. Spontaneous resolution of EP with beta hCG < 1,000 mUl/ml, plasmatic progesterone < 5 ng/ml had 74% success rate. Methotrexate (MTX) injection is the common Medical treatment of EP with beta hCG < or = 5,000 mUl/ml and mean diameter of EP < or = 3 cm: a single dose of 50 mg/m2 or 1 mg/kg intramuscular injection had 93.4% success rate; local injection under sonographic control (1 mg/kg) had 80.2% success rate. The surgical conservative treatment by laparoscopy had 94% success rate.

[Isolated splenic metastasis of ovarian endometrioid carcinoma. A clinical case]. / Metastasi splenica isolata di un carcinoma endometrioide dell'ovaio. Caso clinico.

The authors describe a case of solitary spleen metastasis of an endometrioid ovarian cancer exceptionally reported in the literature. The follow-up of the patient is 12 months without recurrence.

[Resectoscopic endometrial ablation in the treatment of recurrent, dysfunctional menometrorrhagia: our experience]. / L'ablazione di endometrio tramite resettoscopio nel trattamento delle menometrorragie recidivanti disfunzionali: nostra esperienza.

UNLABELLED: Advances in hysteroscopic surgery provide additional options to hysterectomy for the treatment of dysfunctional uterine bleeding resistant to medical therapy and multiple curettages. Two techniques are now available: (a) Resectoscopic endometrial ablation. (b) Electrocoagulation or laser photovaporisation of endometrium. 52 patients underwent resectoscopic endometrial ablation at the Gynaecology Department of Parma University from January 1991 to April 1993. All patients suffered from dysfunctional uterine bleeding without atypical histologic findings on endometrial biopsies and had a normal shaped uterine cavity. 41 patients were subsequently contacted for follow-up. Follow-up period ranged from a minimum of 3 months to a maximum of 24 months. 78.1% of the patients reported a satisfactory outcome (amenorrhea or decreased menstrual flow). No operative complication occurred. Post operative complications included one case of hematometra. CONCLUSIONS: resectoscopic endometrial ablation is an advantageous technique but our follow-up period is relatively short and long term sequelae have yet to be determined.