Enantiorecognition of planar "metallocenic" chirality by a nitrile hydratase/amidase bienzymatic system.

For the first time, a bacterial strain expressing a nitrile hydratase/amidase activity was able to recognize a planar element of chirality: Rhodococcus rhodochrous PA-34 whole cells catalysed with a high level of enantioselectivity the biotransformation of a novel nitrile ferrocene derivative into its corresponding amide and/or acid. An important parameter in the enzymatic recognition is the choice of the inducer selected for the bacterial growth phase.

Multianalytic study on cervical HPV lesions.

Nowadays new sophisticated techniques of molecular biology based on the principles of hybridization between nucleic acids, allow a correct diagnosis of genital HPV infection. In the present paper, beside traditional diagnostic methods, we used In Situ Hybridization (ISH) and Polymerase Chain Reaction (PCR) to detect the presence of HPV types 6, 11, 16, 18, 31 and 33. We tested ten patients affected by cervical lesions of high histological atypias associated with HPV, who underwent surgical conization. Types 6 and 11, at low risk of evolution, are less frequent than 31 and 33, at medium grade of evolution, and than 16 and 18 which are at high risk of evolution.

A role for tumor markers in therapeutic decisions after chemotherapy induction of objective remission in stage III epithelial ovarian neoplasms.

The management of advanced stage ovarian carcinomas is presently based on initial surgical debulking, multiple drug chemotherapy including cisplatinum, second-look laparotomy. Such an aggressive approach has improved objective response rates and expected survival time, but no dramatic change has been demonstrated as for definitive cure percentages. Many Authors have attempted to turn an optimal objective response to chemotherapy (no residual or minimal residual disease at second-look) into a definitive cure with irradiation. Some reports show satisfactory results, but a high incidence of bowel obstructive complications has been demonstrated, probably due to multiple surgical manipulations before radiotherapy. A reliable diagnostic tool, that could help to avoid the second-look laparotomy (whose inherent role in improving survival is not assessed) should be therefore useful. The possible role of serum tumor markers determinations, for this purpose, is here discussed on the ground of a series of 20 patients affected by stage III ovarian carcinoma. Following this experience, a valuable role seems attributable to CA 125 in monitoring tumor response. Patients achieving values under 35 U/ml before second-look laparotomy showed tumor residuals in the range O-microscopic- less than 1 cm., that is, neoplastic localizations reliable for consolidation radiation therapy.

Gonadotropin, prolactin and thyrotropin pituitary secretion after exogenous dehydroepiandrosterone-sulfate administration in normal women.

The effect of exogenous dehydroepiandrosterone-sulfate (DHAS) on luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL) and thyroid-stimulating hormone (TSH) pituitary secretion was studied in 8 normal women during the early follicular phase. The plasma levels of these hormones were evaluated after gonadotropin-releasing hormone (GnRH)/thyrotropin-releasing hormone (TRH) stimulation performed after placebo or after 30 mg DHAS i.v. administration. The half-life of DHAS was also calculated on two subjects; two main components of decay were detected with half-times of 0.73-1.08 and 23.1-28.8 h. The results show an adequate response of all hormones to GnRH or TRH tests which was not significantly modified, in the case of LH, FSH and PRL, when performed in the presence of high levels of DHAS. However, the TSH response to TRH was significantly less suppressed (p less than 0.05) (39%) after DHAS administration than during repeated TRH stimulation without DHAS (51%). The data support the hypothesis that DHAS does not affect LH, FSH and PRL secretion, while TSH seemed to be partially influenced.

Evaluation of tumor-associated trypsin inhibitor (TATI) in women with benign and malignant gynecological disease.

Tumor-associated trypsin inhibitor (TATI) was assayed in healthy subjects and in women with benign and malignant gynecological diseases. Significantly lower levels were found in boys than in healthy adult subjects. No variations in level were evident over the course of a 24 h period. At a cut-off level of 20 micrograms/l elevated concentrations were found in 42%, 11.4% and 19% of women with ovarian, endometrial and cervical neoplasia, respectively. In patients with ovarian tumors TATI level were elevated both in mucinous and serous tumors. TATI does not seem to be useful for diagnosis of uterine tumors, but could have a specific place in the study and management of ovarian tumors, in which serum concentrations can reach levels 100-200 micrograms/l. In the other gynecological diseases maximum levels of 30-40 micrograms/l were observed.

Different dopaminergic control of plasma luteinizing hormone, follicle-stimulating hormone and prolactin in ovulatory and postmenopausal women: effect of ovariectomy.

The main aim of this study was to evaluate the effect of dopamine infusion on plasma luteinizing hormone (LH), follicle-stimulating hormone, (FSH) and prolactin (PRL) after acute (1 week postovariectomy) and chronic (postmenopausal women) estrogen withdrawal. We also studied the same group of postmenopausal women after ovariectomy to evaluate the possible influence of other gonadal factors on the endocrine effects of dopamine. In order to have a further indication of neuroendocrine dopamine activity on pituitary secretions, we measured the change in plasma LH, FSH and PRL after the administration of metoclopramide, a dopamine receptor antagonist. Our findings confirm that in fertile women dopamine infusion inhibits plasma LH and FSH levels and show that 1 week after ovariectomy the LH decrease during dopamine administration is still present whereas the FSH decrease is not. In all groups of patients, dopamine significantly inhibited plasma PRL levels. Metoclopramide increased plasma LH levels in reproductive-age women before ovariectomy, but not in postmenopausal women. Plasma FSH levels did not change in any group and PRL levels increased after metoclopramide administration in all subjects. The present findings show that dopamine regulation of LH is impaired in long-term menopause, but not shortly after ovariectomy. These changes in LH control are not followed by similar changes in PRL secretion, which remains under tonic inhibitory regulation by dopamine. The different behavior of LH and FSH after ovariectomy and in postmenopause shows the independence of LH and FSH regulation.

CA 125 and CA 19-9 in peritoneal, cyst and amniotic fluids.

The concentrations of the tumour markers CA 125 and CA 19-9 were determined in peritoneal, cyst and amniotic fluids, with particular attention being paid to certain reliability criteria of the assay methods. The antigens were measured in undiluted samples and after several dilutions. A recovery test was also performed and protein content evaluated. The results show high levels of CA 125 in all fluids; in descending order of concentration: amniotic (2376-3891 U ml-1), peritoneal (379-4040 U ml-1) and cyst fluid (124-466 U ml-1). Amniotic, peritoneal and cyst fluid concentrations of CA 19-9 were found to be 314-1008 U ml-1, 26.7-2182 U ml-1 and 226-2988 U ml-1, respectively. Recovery was between 80 and 100% for all fluids. CA 125 was easily assayable in all fluids, except amniotic and peritoneal which required dilution even of the samples which fell within the range of the standard curve before dilution. The presence of CA 125 and CA 19-9 in amniotic and cyst fluids emphasizes the non-specificity of these molecules and suggests caution in the interpretation of the results.

Plasma and cyst fluid levels of delta 5 and delta 4 steroid hormones in women with gross cystic breast disease.

delta 5 and delta 4 steroid levels were studied in the plasma and cyst fluid of women with gross cystic breast disease (GCBD). In luteal phase a significant increase in plasma levels (mean +/- SEM) of DHA (11.2 +/- 2.4 ng/ml), DHAS (1.45 +/- 0.6 micrograms/ml) and cortisol 277 +/- 15.7 ng/ml) was found; in follicular phase the mean levels were 4.09 +/- 0.47 ng/ml for DHA, 0.65 +/- 0.08 microgram/ml for DHAS and 190 +/- 46.3 micrograms/ml for cortisol. The DHA/DHAS and cortisol/androstenedione ratios were significantly higher in the plasma and lower in the cyst fluid of GCBD patients, than in the plasma of controls; the androstenedione/DHA ratio was higher in the cyst fluid than in the plasma of controls. The hormonal situation of the GCBD patients thus differed from that of the controls both in the plasma and cyst fluid, particularly as regards the delta 5 steroids.

Prolactin changes after administration of agonist and antagonist dopaminergic drugs in puerperal women.

Prolactin (PRL) is an anterior pituitary hormone which plays a large part in the reproductive function of mammals. Its only well-documented effect in humans is that of initiating and maintaining lactation. Among hypothalamic neurotransmitters regulating the anterior pituitary function, dopamine (DA) is currently considered to correspond to the PRL-inhibiting factor. The central control mechanisms which induce high PRL levels in puerperal women are not well understood. To study DA tonus in puerperium we tested plasma PRL levels in different groups of puerperal subjects (6 per group) after acute administration of direct or indirect DA agonists or placebo: DA, L-dopa (a DA precursor), L-dopa plus carbidopa (a peripheral dopa-decarboxylase inhibitor), nomifensine (a DA-releasing and blocking or reuptake agent) and amphetamine (a DA releaser). The same tests with the same drug doses were performed on groups of healthy volunteers. A consistent reduction in plasma PRL levels after both direct and indirect DA agonist drugs compared to placebo was evident in puerperal and in control women. A different trend was only observed with the use of DA and amphetamine in puerperal subjects, who, unlike controls, failed to show a rebound in plasma PRL levels after the termination of drug infusion. These findings support the view that the inhibitory control of tuberoinfundibular neurons over PRL secretion is maintained in puerperium and changes in the affinity of DA receptors are related to the endocrine milieu which occurs during gestation.

Prolactin secretion after surgery or bromocriptine treatment of prolactinoma.

In prolactinoma patients, the unresponsiveness of prolactin (PRL) to dynamic tests with thyrotropin-releasing hormone (TRH) or dopamine agonist or antagonist drugs suggests that the disease is caused by the failure of central dopaminergic inhibition. The absence of PRL secretory response to dopamine or TRH in prolactinoma patients also may be an effect of the disease. Twenty-six women diagnosed by dynamic tests and radiologic examination as having PRL-secreting adenoma were treated surgically (15) or with bromocriptine (11) and their condition was evaluated one to three years later by TRH, nomifensine, and domperidone tests. Basal PRL levels decreased after surgery and bromocriptine treatment. At the time of the follow-up study, PRL levels were elevated in six of the 15 surgically treated patients and in six of the 11 treated with bromocriptine. Thyrotropin-releasing hormone, nomifensine, and domperidone produced standard PRL responses in normoprolactinemic patients but not in hyperprolactinemic patients. These results indicate that the alteration of TRH or dopaminergic receptors in the regulation of PRL secretion in prolactinoma is related to the disease and disappears when the tumor is removed or treated successfully with bromocriptine.

Domperidone in defective and insufficient lactation.

The clinical use of anti-dopaminergic drugs to stimulate plasma PRL levels, to induce lactogenesis and maintain an adequate lactation has been widely suggested, taking into consideration the main inhibitory role of hypothalamic dopamine on PRL secretion. We therefore studied the effects of domperidone (DOM), a direct anti-dopaminergic drug with a low tendency to be secreted in the milk and which does not cross the blood-brain barrier, on inducing lactogenesis in 8 puerperal women with a history of defective lactogenesis (group A) and inducing galactopoiesis in 9 puerperal women who showed 2 weeks after delivery an insufficient lactation (group B). A placebo treatment was performed in 7 and 8 puerperal women with the same characteristics of group A and B, respectively. PRL plasma levels were assayed in basal conditions and after suckling from the 2nd to the 5th day of puerperium in group A and through a 10-day treatment in group B. In both groups domperidone-treated subjects always showed baseline PRL levels and daily milk yield significantly higher than those of the placebo group (P less than 0.01). The lack of any side-effects and the positive results suggest a high usefulness of such a drug in inducing and/or maintaining successful breast feeding, which is at present considered so important for a healthy development of infants.

Central opioid receptor activity in patients affected by puerperal, idiopathic and tumoral hyperprolactinemia.

The aim of our study was to evaluate the activity of opioid receptors involved in the regulation of prolactin secretion in different kinds of hyperprolactinemic states (puerperal, idiopathic and tumoral subjects). Prolactin plasma level changes were measured after the acute administration of a met-enkephalin synthetic analogue. FK 33-824, and placebo. The prolactin-releasing effect of this drug, evident in control subjects, was blunted in patients affected by idiopathic hyperprolactinemia, and absent in puerperal and tumoral patients. These results indicate that hyperprolactinemia affects the activity of central opioid receptors, with the most evident effects in patients with highest prolactin plasma levels.

[Changes in plasma concentrations of adrenal androgens in patients with chronic renal insufficiency]. / Alterate concentrazioni plasmatiche degli androgeni surrenalici in pazienti con insufficienza renale cronica (IRC).

The present study points out that the CHRONIC RENAL FAILURE (CRF) represents a situation of decreased adrenal function: at least for delta 5 Steroids which are markedly reduced when compared with normal subjects. Peripheral plasma levels of Pregnenolone (delta 5 P) ranged in CRF between 190 and 860 pg/ml; Dehydroepiandrosterone-sulphate (DHA-S) 0.1-2.2 ng/ml and Dehydroepiandrosterone (DHA) 200-3100 pg/ml. Cortisol was in the normal range or slightly elevated (70-175 ng/ml). A significant correlation between basal concentration and the time of dialysis was observed. It is interesting to notice how the phase levels of delta 5 P, DHA-S and DHA are reduced ad from the beginning of the haemodialysis treatment and that during such a treatment a further progressive decrease occurs reaching concentrations with are similar to those found in pre-puberty. This phenomenon appears to be the evidence of a progressive metabolic involution of the adrenal gland due to the exhaustion of enzymatic activities and of receptorial structures. Furthermore, these results suggest speculation on interrelationship between adrenal and gonadal activity in these patients.

[Various aspects of dopaminergic function in patients with prolactin-secreting hypophyseal adenoma]. / Vari aspetti della funzione dopaminergica in pazienti affetti da adenoma ipofisario PRL-secernente.

Controversial data have been published in these last years on the dopaminergic tonus of tubero infundibular (TIDA) neurons in hyperprolactinemic subjects. Some authors retain that L-Dopa (LD) stimulation test after pretreatment with Carbidopa (CD), a dopa decarboxylase inhibitor substance, may reveal the presence of a central Dopamine (DA) defect in patient with prolactinoma. To elucidate the reason of so different activity of DA central tonus in subjects with prolactinoma, the effects of Nomifensine (NOM), an indirect DA agonist, DOM, Carbidopa/L-Dopa (CD/LD) and LD, were studied in 26 prolactinoma patients, (basal Prl levels 50 to 280 ng/ml). The patients (24-39 years) were characterized by secondary amenorrhea with sella turcica enlargement at hypocicloidal polytomography. The NOM administration resulted unable to reduce Prl plasma levels in all the patients. On the basis of Prl response to CD/LD administration the patients were subdivided in two groups: group A, which showed a significant decrease of Prl plasma levels and group B, who did not show any significant change. LD test induced a significant Prl decrease in all patients with more evident response in these of group A. DOM administration induced a Prl rise in patients of group A, but failed to change significantly Prl levels in group B subjects. These results confirming the high validity of NOM inhibiting test in the diagnosis of tumoural hyperprolactinemic states, reveal contradictory responses to CD/LD, LD and DOM, with sustain the existence of 2 sub-group of Prolactinomas: with or without a maintained DA central tonus supporting the possibility of different etiopathogenetical factors in inducing a tumoural hyperprolactinemic states.

[Hyperprolactinemia in chronic renal insufficiency: an interesting model of hypothalamo-hypophyseal dysfunction]. / L'iperprolattinemia nella insufficienza renale cronica (IRC): un interessante modello di disfunzione ipotalamo - ipofisaria.

It is note that hyperprolactinemia is frequently associated to chronic renal failure (CRF). The etiopathogenesis of this endocrine disorder is not clearly understood, trying to evaluate the possible hypothalamic-pituitary cause we have evaluated the Prl levels under some pharmacological tests: TRH (200 mcg i.v.),Domperidone (DOM)(10 mg i.v.),Nomifensine (NOM)(200 mg p.o.)and Bromocriptine (BRC)(2.5 mg p.o.) in 3 groups of patients: CRF(8 cases), dysfunctional (8 cases) and tumoral (9 cases) hyperPrl. Prl plasma levels have been evaluated by RIA (kits,Biodata,Roma). In patients affected by CRF either direct (BRC) than indirect (NOM) agonist acting dopaminergic drugs failed to induce a reduction in Prl plasma levels, like to tumoral patients for NOM, and in contrast to the significant decrease after BRC (-70%) in tumoral and (-74%) in dysfunctional and NOM (-50%) observed in dysfunctional patients. CRF and dysfunctional patients showed significant response to TRH and DOM, being prolactinoma bearing patients unresponsive to these tests. These results show an hyporesponsivity of pituitary Prl secreting cells to dopaminergic control suggesting the existence of central etiopathological factor in inducing hyperprolactinemia in CRF patients.

Puerperal breast feeding does not stimulate circulating opioids in humans.

ACTH, beta lipotropin (beta LPH), beta endorphin (beta EP), Prolactin (PRL) and Cortisol were measured in the first five days of puerperium at 9:00, before and 30 minutes after suckling, in 7 healthy lactating women. With the exception of PRL plasma levels which decline, although remaining at high concentrations during the observation period, all the other parameters showed a sudden fall from the high levels found at delivery, reaching stable normal levels (beta LPH, beta EP, Cortisol) or concentrations which were 50% lower than normal (ACTH), from the second day of puerperium. Suckling confirms its capacity to release plasma PRL, while all the other indices remain unmodified. Despite the experimental evidence that serotoninergic neurons are involved both in the PRL response to suckling and the circadian rhythmicity and stress response of proopiocortin-related peptides, the present results suggest that breast feeding is not a stressful situation, as no typical proopiocortin-related peptide response is evident, and that the activation of PRL release is related to stimulation of serotoninergic neurons which differ from those involved in the control of proopiocortin-related peptide secretion.

Opioid plasma levels during labour.

6 healthy pregnant women, monitored with external cardiotocography, were studied. Blood samples were collected hourly throughout labour until delivery of the fetus and placenta, and again on the 5th day of puerperium. beta-Lipotropin (beta LPH) and beta-endorphin (beta EP) were determined by specific radioimmunoassays in each sample, after silicic acid plasma extraction and G-75 column chromatography. Both opioid plasma levels rise progressively during labour, reaching their highest values 1 h before delivery (beta LPH: 296.0 +/- 60.2 pg/ml; beta EP 106.2 +/- 40.6 pg/ml) as compared with values of 157.0 +/- 35.4 (beta LPH) and 57.0 +/- 7.3 pg/ml (beta EP) (mean +/- SE) at the beginning of labour. With the exception of 1 case, a significant correlation was observed between beta LPH and beta EP plasma levels. The two opioid plasma levels decrease after delivery of the placenta (272.5 +/- 59.6 and 118.2 +/- 68.4 pg/ml) and subsequently decrease further to levels of 105.6 +/- 46.4 (beta LPH) and 33.7 +/- 16.5 (beta RP) on the 5th day of puerperium. beta LPH and beta EP plasma levels showed a significant correlation throughout labour with the 'uterine contractility force/hour', which was calculated by the addition of tocographic areas of each uterine contraction at 1-hour intervals. These data demonstrate that the increase in plasma opioid concentrations during labor is directly related to the number and intensity of uterine contractions.