L-Carnitine counteracts in vitro fructose-induced hepatic steatosis through targeting oxidative stress markers.

PURPOSE: Nonalcoholic fatty liver disease (NAFLD) is defined by excessive lipid accumulation in the liver and involves an ample spectrum of liver diseases, ranging from simple uncomplicated steatosis to cirrhosis and hepatocellular carcinoma. Accumulating evidence demonstrates that high fructose intake enhances NAFLD development and progression promoting inhibition of mitochondrial ß-oxidation of long-chain fatty acids and oxidative damages. L-Carnitine (LC), involved in ß-oxidation, has been used to reduce obesity caused by high-fat diet, which is beneficial to ameliorating fatty liver diseases. Moreover, in the recent years, various studies have established LC anti-oxidative proprieties. The objective of this study was to elucidate primarily the underlying anti-oxidative mechanisms of LC in an in vitro model of fructose-induced liver steatosis. METHODS: Human hepatoma HepG2 cells were maintained in medium supplemented with LC (5 mM LC) with or without 5 mM fructose (F) for 48 h and 72 h. In control cells, LC or F was not added to medium. Fat deposition, anti-oxidative, and mitochondrial homeostasis were investigated. RESULTS: LC supplementation decreased the intracellular lipid deposition enhancing AMPK activation. However, compound C (AMPK inhibitor-10 µM), significantly abolished LC benefits in F condition. Moreover, LC, increasing PGC1 α expression, ameliorates mitochondrial damage-F induced. Above all, LC reduced ROS production and simultaneously increased protein content of antioxidant factors, SOD2 and Nrf2. CONCLUSION: Our data seemed to show that LC attenuate fructose-mediated lipid accumulation through AMPK activation. Moreover, LC counteracts mitochondrial damages and reactive oxygen species production restoring antioxidant cellular machine. These findings provide new insights into LC role as an AMPK activator and anti-oxidative molecule in NAFLD.

A new hydrogel for the extended and complete prednisolone release in the GI tract.

The issue of incomplete release of poorly soluble drugs from sustained-release oral formulations is addressed using prednisolone (PDS) as the model drug and a novel highly swelling hydrogel as the rate-controlling material. The hydrogel was formed by heating N-carboxymethylchitosan (CMC) to 80 degrees C for 24 h. Swelling, alkalimetry, FTIR, DSC, and solid-state NMR studies showed that the treatment produced physical crosslinking, i.e., polymer chain entanglement. A controlled-release system was prepared by coating an inert compacted support of ethylcellulose (50 mg; diameter, 6 mm) with a CMC layer containing dispersed PDS powder (10-50 microm). The system was heated to crosslink the CMC coating, then drug release to simulated GI fluids was studied in vitro. The drug release pattern and term were modulated via the layer mass (LM) (10 or 14 mg cm(-2)) and/or the drug-polymer wt ratio (D/P) (1:5 or 2:5). The rate parameter, K, and the time exponent, n, of the Peppas equation were: K=26.6+/-0.3 h(-n), n = 0.78+/-0.02 (LM, 10 mg cm(-2); D/P, 1:5); K = 24.7+/-0.7 h(-n), n = 0.56+/-0.02 (LM, 14 mg cm(-2); D/P, 1:5); K = 20.7+/-0.3 h(-n), n = 0.76+/-0.01 (LM, 10 mg cm(-2); D/P, 2:5). Hydrogel swelling was faster than drug release. This was controlled, in a first stage, by drug dissolution-diffusion in the swollen gel, and subsequently, by diffusion. The drug release rate was unaffected by the GI pH variations, and slightly affected by the environmental hydrodynamics. The system promises an extended and complete release of poorly soluble drugs in the GI tract.

Seasonal onset of the menopause.

OBJECTIVES: A seasonal rhythm of reproduction is evident in humans. Herein it was investigated whether also the cessation of woman's fertile life follows a seasonal rhythm. METHODS: A retrospective study was performed on 2436 women in postmenopause for more than 12 months, in our menopause centres. Time of menopause was stratified for month and season. The variation was compared to the seasonal rhythm of 14,310 conceptions. RESULTS: The onset of menopause was more frequent (p<0.0001) in winter (32.5%) than in spring (20.8%), autumn (20.3%) and summer (26.2%), in which a minor peak was also observed (p<0.0001 vs. spring and autumn). The two peaks were temporally coincident with the transitions between the high to low and low to high rate of conceptions. CONCLUSIONS: The present data show that in women, like reproduction also the onset of menopause shows a seasonal modulation.

Relative influence of menopausal status, age, and body mass index on blood pressure.

The objective of this study was to assess the influence of menopausal status on blood pressure levels in a cross-sectional population study in the setting of a preventive health care program clinically conducted in Ferrara, Italy. The patients were 2397 healthy women 35 to 65 years old (489 in premenopause, 847 in perimenopause, 887 in spontaneous menopause, and 174 in surgical menopause); subgroups were also obtained with increasing duration of menopause (1 to > 5 years) matched with premenopausal and perimenopausal women by chronological age at onset of menopause. Measurements of sphygmomanometric blood pressure, age, age at menopause, duration of menopause, and body mass index were made. Postmenopausal women had higher blood pressure than premenopausal and perimenopausal subjects. After adjustment for body mass index, the blood pressure changes with menopausal status were still significant, but not after correction by age. The increased risk of hypertension of postmenopausal women also lost statistical significance after adjustment for age and body mass index. At multivariate analysis, blood pressure showed no significant relation with the duration of menopause, whereas age was a significant covariant for systolic blood pressure, and body mass index for systolic and diastolic blood pressure. A transient initial rise in blood pressure and body mass index was detected in surgical but not in spontaneous menopause. Blood pressure rises after menopause appear to be due more to increased body mass index and aging than ovarian failure per se.

Influence of estrogens on serum free fatty acid levels in women.

A relationship between plasma levels of FFA and incidence of hormone-dependent breast cancer has been suggested. This observation has drawn our attention to possible complementary actions of ovarian steroids on circulating FFA levels. Measurements taken in normal women during the menstrual cycle and in ovariectomized women with and without estrogen replacement therapy demonstrate that 1) levels of FFA present in serum are lower during the luteal phase than during the follicular phase; 2) levels of FFA are significantly higher after ovariectomy; and 3) these are again reduced by substitutive estrogen therapy.

Human vascular kinin receptors of the B2 type characterized by radioligand binding.

1. The human umbilical vein responds to bradykinin (BK) with contractions that are mediated by B2 receptors. In the present study, the corresponding vascular smooth muscle B2 binding sites have been investigated. 2. [3H]-BK, a full agonist labelled ligand, was used to demonstrate a single binding site giving a Kd value of 0.51+/-0.02 nM and a Bmax of 24+/-1 fmol mg(-1) protein. Scatchard plots were linear (r=0.98) in the 0.05-5 nM range of concentrations. Non-specific binding was found to be 30% of total binding. 3. Competition binding curves gave the following order of potency for various B2 receptor agonists: BK-[Hyp3]-BK > or = Lys-BK >> [Aib7]-BK >>> [desArg9]-BK, which is typical of B2 receptors. There was no binding to B1 receptors since the selective B1 receptor ligand, Lys-[desArg9]BK was inactive up to 10 microM (n=4). 4. Characterization of the binding site with antagonists, performed with three chemically distinct series of peptide and non-peptide compounds, revealed a high affinity of Hoe 140 (D-Arg-[Hyp3,Thi5,D-Tic7,Oic8]-BK) (Ki 0.17 nM; n=4) which was more potent that FR 173657 ([(E)-3-(6-acetamido-3-pyridyl)-N-[N-[2,4-dichloro-3-[(2-methyl-8-quinol inyl)oxymethyl]phenyl]-N-methylaminocarbonylmethyl] acrylamide]) (Ki 1.94 nM; n=4), D-Arg-[Hyp3,D-Phe7,Leu8]-BK (Ki 256 nM; n=4) and Win 64338 (phosphonium, [[4[[2[[bis(cyclohexylamino)methylene]amino]-3-(2-naphthalenyl)-1-oxopro pyl]amino]phenyl]methyl]tributyl, chloride, monohydrochloride) (Ki 1,450 nM; n=4). 5. The present study describes and characterises B2 receptor binding sites in the vascular smooth muscle of the human umbilical vein. The binding assay appears to be suitable for studying new agonists or antagonists designed to activate or block the B2 receptor class that mediate the majority of the physiopathological effects of kinins in man.

Ovarian steroids modulate the action of calcitonin in women.

Slow i.v. infusion of salmon calcitonin into normal women led to a transient increase in plasma concentrations of cyclic AMP and to a decrease in plasma calcium, which was maintained for at least 2 h. These responses were significantly reduced in the same patients after hysterectomy and ovariectomy. In contrast, maintenance of the ovaries at the time of surgery, or substitutive hormonal therapy, seemed to restore the sensitivity to exogenous calcitonin, suggesting a permissive role of ovarian steroids in the effect of calcitonin. The most likely explanation for this phenomenon is that these hormones influence either the number of calcitonin receptors or the coupling of the receptors to the functional unit of bone adenylate cyclase.

Characterization of FR 172357, a new non-peptide bradykinin B(2) receptor antagonist, in human, pig and rabbit preparations.

FR 172357, a new non-peptide antagonist of the kinin B(2) receptor was tested in three isolated vessels, the human umbilical vein, the rabbit jugular vein, and the pig coronary artery, to evaluate its antagonistic activities against bradykinin. FR 172357 displaced to the right the concentration-response curves of bradykinin. The displacements were parallel to the controls without reduction of the maximum effect in the human umbilical vein and in the rabbit jugular vein, but not in the pig coronary artery. Schild plots confirmed that FR 172357 acts as a competitive antagonist in the human umbilical vein (pA(2) 8.65) and in the rabbit jugular vein (pA(2) 9. 07), and as a non-competitive antagonist in the pig coronary artery (pK(B) 10.14). FR 172357 is selective for the kinin B(2) receptor since it does not influence the effects of Lys-des-Arg(9)-bradykinin in the human umbilical vein, in the rabbit aorta, and in the pig renal vein. It is specific because it does not affect the contractions induced by angiotensin II, noradrenaline, 5-hydroxytryptamine, or endothelin-1 in the human umbilical vein. It, however, interacts with the tachykinin NK(1) receptor of the rabbit jugular vein and pig coronary artery. Compared to other bradykinin B(2) receptor antagonists, FR 172357 emerges as a very potent compound, which may represent a choice for experimental (and clinical?) applications.

Influence of spontaneous and surgical menopause on atherogenic metabolic risk.

We evaluated serum total-, LDL-, HDL-, cholesterol and triglycerides in 858 women. Sixty-seven were in premenopause, 307 in perimenopause, 326 in spontaneous menopause, and 158 women underwent hysterectomy. Of these, 101 had bilateral ovariectomy, 24 with unilateral ovariectomy and 33 with retention of both ovaries. After correcting for chronological age and body mass index by variance analysis, it was found that total-, LDL- and HDL-cholesterol significantly increased during menopause without changes in the total cholesterol/HDL-cholesterol, LDL-cholesterol/HDL-cholesterol and triglycerides/HDL-cholesterol ratio. The changes gradually occurred during the months preceding spontaneous cessation of menses. Atherogenic metabolic risk (as relative risk of high total and LDL-cholesterol, estimated by logistic regression analysis) significantly increased in all groups of women in reference to premenopause. The risk observed in surgical menopause with bilateral ovariectomy was higher than in spontaneous menopause, and conservation of ovaries in hysterectomized women seems to protect only partially against such an increase.

Blood prolactin levels: influence of age, menstrual cycle and oral contraceptives.

The relationships between the serum levels of estradiol, progesterone and prolactin have been analysed in normal women in relation to age, to menstrual cycle, and in oral contraceptives users. We have been able to detect significant differences between the prolactinemia values during the cycle in different age groups as well as following oral contraceptives. A word of caution is presented for the evaluation of the prolactin blood level which should always be related to the physiological phase of the cycle and to the age of the subjects.

LOH of chromosome 6q compared with LOH of 17q and 18q in ovarian cancers: relationship to p53 expression and clinicopathological findings.

In 41 ovarian epithelial tumors (7 borderline and 34 invasive), loss of heterozygosity (LOH) of chromosomes 6q, 17q, and 18q was examined using 4 microsatellite markers: ER (6q 25-1), BRCA1 (17q21), DCC (18q21), and D18S58 (18q23). The LOH was compared with clinicopathological findings, including p53 and ER expression. In borderline tumors, LOH and p53 expression were never found, while in invasive carcinomas LOH and p53 were found in 71% and 59% of cases, respectively. In particular, in invasive carcinomas 6q LOH represented a marker distinguishing two groups of tumors; those with 6q LOH were only of serous histotype and at advanced stages (III/IV). No significant difference was found for any of genes in 5-year survival of the patients. No correlation was found between ER expression and ER LOH, as well as between biological aggressiveness and 17q and/or 18q LOH. We conclude that p53 and LOH of the investigated loci distinguish borderline from invasive ovarian carcinomas; moreover, the comparison of these results with clinicopathological parameters suggests that the presence of 6q LOH may be a factor accounting for greater biologic aggressiveness independent of the histologic subtype.

Haematocervix during hormonal replacement therapy.

A postmenopausal woman, receiving cyclic hormonal therapy, presented to our clinic with acute pelvic pain, absence of withdrawal bleeding and clinical signs of vagal stimulation (nausea and vomiting). Vaginal exploration and ultrasonographic imaging accounted for haematocervix, which was confirmed by surgical blood evacuation.

The menopausal transition: a dynamic approach to the pathogenesis of neurovegetative complaints.

In our cross-sectional study we investigated the separate influence of three main factors, namely menopausal and estrogen status, and chronological age, on ten neurovegetative climacteric complaints reported in the scale of Kupperman et al. A multivariate statistical analysis was performed by a multivariate statistical approach on 1161 untreated women seen at the Menopause Center of the Ferrara University Hospital. Ninety women (age range, 41-54 years) were premenopausal; 492 women (age range, 38-55 years) were perimenopausal with irregular periods or amenorrhea for less than 12 months; 468 women (age range, 41-69 years) had a spontaneous menopause (age range, 37-66 years); 111 had had hysterectomy with bilateral ovariectomy while still regularly menstruating. Serum estrone was used as the indicator of the patients' estrogen status. A clear positive trend was demonstrated between menopausal status and the prevalence of depression, hot flushes, insomnia and joint pain. However, only the prevalence of hot flushes amongst these four symptoms was significantly related with the climacteric estrogen decline (beta = -0.006, P = 0.001). Moreover, menopausal status appeared to influence the intensity of fatigue, hot flushes, insomnia and paresthesia. Age was found to significantly (P = 0.053) co-vary only with the intensity of the hot flushes, with a positive relation (beta = 0.092, r = 0.104, P = 0.003), whereas estrone values did not significantly co-vary with any symptom. Furthermore, while neurovegetative symptoms are largely present also in the absence of hot flushes, when these latter are present, they exacerbate both the intensity and the prevalence of all the other symptoms.(ABSTRACT TRUNCATED AT 250 WORDS)

Modification of number and of affinity of endometrial EGF receptors during the menstrual cycle.

We have analysed the distribution and the binding properties of the epidermal growth factor (EGF) receptor in human endometrium during the menstrual cycle to validate the proposal that steroids affect the growth rate of target cells via growth factor-related pathways. The total number of EGF receptors and their affinity for the iodinate exogenous ligand are significantly higher during the proliferative than the secretory phase; the binding kinetics appear to be hyperbolic and sigmoid during the proliferative and the secretory phase, respectively. These results suggest that endogenous sex steroids can regulate the levels and the properties of EGF receptors in vivo, and support the possibility that receptor-coupled events may play a role in estrogen-stimulated growth.

Serious hematoma of the vulva from a bicycle accident. A case report.

BACKGROUND: Vulvar hematomas may be caused by obstetric, athletic or sexual trauma or by car or bicycle accidents. CASE: A 29-year-old woman presented with a serious vulvar hematoma caused by a straddle bicycle accident. Monitoring of the hematologic parameters showed progressive anemia resulting from silent bleeding, which required surgical evacuation of the hematoma and complete hemostasis. CONCLUSION: The rich vulvar vascular network may be easily damaged by contusive frontal impacts, which crush the vulvar tissues against the osseous planes. Management of vulvar hematomas may range from conservative to surgical. Drainage and incision are advised in more severe cases to reduce infective complications and hospitalization.

Open study of topical 0.025% tretinoin in the treatment of vulvar lichen sclerosus. One year of therapy.

OBJECTIVE: To study the use of topical tretinoin for treating vulvar lichen sclerosus. STUDY DESIGN: An open, uncontrolled clinical study on 22 patients affected by histologically confirmed vulvar lichen sclerosus. Topical 0.025% tretinoin was applied once a day, five days a week, for one year. Clinical and histologic parameters were evaluated before and after therapy, and statistical analysis was performed. RESULTS: Symptoms, gross appearance and histopathologic features improved in a highly significant manner (P < .001). Cutaneous side effects were observed but rapidly disappeared, and no patient left the study for this reason. Maintenance of results was observed at the 4-13-month follow-up visits. CONCLUSION: Topical tretinoin seems feasible for use in the topical treatment of vulvar lichen sclerosus.

Risk factors in vulvar carcinoma recurrences.

18 patients with invasive vulvar carcinoma treated by radical vulvectomy and bilateral groin lymphadenectomy have been evaluated. An 18% of local and 22% of pelvic or distant recurrences occurred. The local recurrences had favorable prognosis, whereas the pelvic or distant recurrences a worse prognosis. The staging, the tumour size, the nodes involvement, the stromal infiltration and the lymphoplasmacytic infiltration, as prognostic factors, have been evaluated. All these factors, except the tumour size, influenced the pelvic or distant recurrences. The pelvic or distant recurrences were contemporaneously associated with mild phlogistic infiltration and depth of invasion > 5 mm.

Surgical treatment of invasive carcinoma of the vulva. A reappraisal.

The Authors studied three groups of patients affected by invasive vulvar carcinoma. The first group (19 cases), treated by radical vulvectomy and bilateral inguinal lymphadenectomy, had a survival rate at 5 years of 89% in Stage I and 56% in Stage II. The second group (9 cases) which presented poor general health conditions, had a survival rate at 4 years of 33% and 14% in Stage I and in Stage II, respectively. The third group of patients (7 cases), who refused any type of treatment, died within 12 months. Nodal involvement influenced survival rate. In fact, regardless of the stage, a survival rate at 5 years of 92% and 26% was seen in patients with negative nodes and positive nodes, respectively. In conclusion, the study confirms that radical surgery is the therapy of choice in advanced carcinoma of the vulva. However, early diagnosis remains the most important agent in reducing the extension of the surgical treatment.

[Cerebral lesions in patients with ovarian cancer]. / Lesioni cerebrali in pazienti con carcinoma ovarico.

The Authors report the appearance of Central Nervous System lesions in three patients previously treated for ovarian carcinoma. In one case (Stage 1) the histological sample found a glioblastoma, in the others (Stage 3) the lesion was the metastases after systemic diffusion of the primary carcinoma. CNS metastases are rare and more frequently occur in advanced ovarian carcinoma. In patients at Stage 1, CNS isolated lesions may be primary tumors.

Elective uterine myomectomy in pregnant women.

One hundred and six pregnant women with uterine myomas were admitted to a clinical protocol for operative or conservative management of myomas. The criteria for decision to submit to surgery were: (i) recurrent pain; (ii) large or rapidly growing myomas; (iii) large or medium myomas located in the lower uterine segment or deforming the placental site. According to these criteria 18 patients underwent myomectomy (operative group) and 88 were conservatively treated (conservative group). Complications and fetoneonatal outcome were monitored. The data were also compared to a control group of 2463 normal pregnant women. No spontaneous abortion occurred in the operated group, while the conservative group had a 13.6% spontaneous abortion rate (control group: 9.3%). The operated group had the highest cesarean section rate (93.7%), but no post-cesarean hysterectomy. The conservative group had a higher cesarean section rate compared with the pregnant women without myomas (34% vs 16.3%, respectively); they also had a 4.5% post-cesarean hysterectomy rate compared with 0.12% in the control group. Fetoneonatal outcome was good both in the conservative and operated group. The only perinatal death regarded a patient with a large myoma who refused surgery and went into premature labour at 32 weeks of gestation. Therefore, our study demonstrates that myomectomy during pregnancy is of advantage when performed early on selected patients.