Role of putrescine in cell proliferation in a colon carcinoma cell line.

OBJECTIVES: Putrescine, the precursor for higher polyamine biosynthesis, is necessary for cell growth in mammals. Ornithine decarboxylase (ODC) activity and polyamine production are increased in neoplastic cells. Using colon cancer cell line derived from a tumor with high metastatic potential (CT-26), our objective was to study the effect of exogenous putrescine on ODC regulation, polyamine metabolism, and cell proliferation. METHODS: Cells cultured with fetal calf serum were exposed to 100, 550, and 1000 microM putrescine for 24 h. RESULTS: Intracellular free putrescine, determined by high-performance liquid chromatography, showed a statistically significant increase in exposed cells compared with controls and a significant correlation with levels of the metabolite present in the medium (r = 0.93; P < 0.001). Bromodeoxyuridine incorporation into newly synthesized DNA, a marker of cell proliferation, showed a statistically significant increase in the three putrescine groups as opposed to the control group. In samples with added aminoguanidine, significant increases in DNA synthesis were observed in the 550- and 1000-microM putrescine groups as opposed to the control group. Spermidine and spermine intracellular contents in all three putrescine-treated groups remained below control levels. No statistical differences in ODC enzymatic activity or ODC mRNA content were observed. Newly incorporated putrescine stimulated colon tumor cell growth. CONCLUSIONS: Because neither enhanced conversion into the higher polyamines nor aminoguanidine inhibition of proliferation was observed, we suggest that this effect can be attributed to the putrescine molecule itself.

[Subarachnoid hemorrhage, cerebral ischemia and endothelin-1]. / Hemorragia subaracnoidea, isquemia cerebral y endotelina-1.

INTRODUCTION: Cerebral vasospasm is involved in the development of delayed ischemic lesions in patients with subarachnoid hemorrhage. We developed an integral theoretical model to explain the pathophysiology of cerebral vasospasm, in which endothelin-1 has a pivotal role in the development of both cerebral vasospasm and delayed ischemic neurological deficits (DIND). OBJECTIVE: The objective of this study is to analyze the relationship between temporal profile of plasma endothelin-1 levels and the development of cerebral vasospasm and DIND. PATIENTS AND METHODS: We analyzed sequentially plasma endothelin-1 levels in 17 patients with aneurysmatic subarachnoid hemorrhage. All the patients had complete clinical and neuroradiological studies. Patients were classified according to Fisher's score. RESULTS: Patients (4 males and 13 females, aged 48.1 +/- 20.3 years) had a good clinical condition (Hunt-Hess < 4, GCS > 10). Two weeks after bleeding, patients had higher plasma endothelin-1 levels than healthy volunteers (p = 0.024). Patients who developed DIND had higher plasma endothelin-1 levels (p = 0.034) and a different evolution (p = 0.0146) than patients without DIND. There is a significant correlation (p = 0.02) between basal plasma endothelin-1 levels and GOS score. Multiple regression analysis shows a significant dependence between plasma endothelin-1 levels and Fisher's score (p = 0.0195), development of DIND (p = 0.0095), and GOS score (p = 0.0319). Logistic regression analysis finds a predictive relation between Fisher's score and plasma endothelin-1 levels for the development of DIND (overall predicted = 74.24%; p = 0.0148). CONCLUSIONS: Plasma endothelin-1 levels are increased in patients after subarachnoid hemorrhage and are associated with the development of cerebral vasospasm and DIND.

Influence of surgical stress and parenteral nutrition on serum leptin concentration.

UNLABELLED: Experimental studies suggest that leptin may be an important metabolic signal for energy regulation. AIM: To assess whether surgical stress produces changes in serum leptin concentration and to investigate and compare the effect of total parenteral nutrition and hypocaloric parenteral nutrition on serum leptin levels. PATIENTS AND METHODS: Twenty-two surgical patients (11 male and 11 female) in need of parenteral nutrition were recruited. Parenteral nutrition was always initiated 24 h after surgical procedure. Group I (n=15) received total parenteral nutrition, while Group II (n=7) were treated with hypocaloric parenteral nutrition. Serum leptin concentration was determined before surgical procedure (day -1), after surgery and before parenteral nutrition was started (day +1), and after 5 days of treatment with parenteral nutrition (day +6). RESULTS: A tendency to increase serum leptin levels was observed after surgical procedure (6.0+/-1.9 vs 9.9+/-2.7 ng/ml;P= 0.07). After starting parenteral nutrition no significant changes on serum leptin concentrations were found in both groups, but a trend to raise serum leptin was observed in Group I (6.2+/-1.7 vs 8.3+/-2.7 ng/ml) whereas a trend to decrease serum leptin was detected in Group II (4.6+/-2.5 vs 1.6+/-0.5 ng/ml). On day +6 an increase of serum leptin and insulin levels was observed in Group I in comparison with Group II (8.3+/-2.7 vs 1.6+/-0.5 ng/ml;P<< 0.05 and 58+/-41 vs 12+/-15 microU/l;P<< 0.05 respectively). Finally, a positive correlation at day +6 between insulin and serum leptin levels was observed (r= 0.66;P<< 0.01). CONCLUSIONS: a) Surgical stress is associated to an increase of serum leptin concentrations; b) Total and hypocaloric parenteral nutrition produces quite different effects on serum leptin levels that could be related to distinct insulin response.

In vitro effects of melatonin on cell proliferation in a colon adenocarcinoma line.

The effect of melatonin on inhibition of cell growth was studied in CT-26, a murine colon carcinoma-derived cell line. Cells growing in exponential phase were exposed to low (10(-7)-10(-10) M) and high doses (1, 2 and 3 x 10(-3) M) of melatonin during 24 h. Synthesis of DNA was measured by 5-bromo-2'-deoxyuridine incorporation. There was no effect at low doses, but a statistically significant correlation was found between the decrease in DNA synthesis and the dose of melatonin used (r = -0.52, P < 0.001). This implied the following percentages of inhibition: 1 mM, 22%; 2 mM, 25%; 3 mM, 47%. Potential cell membrane damage by high doses of melatonin was investigated by lactate dehydrogenase measurement and no significant levels were observed. Analysis with a single saturation technique showed no detectable oestradiol receptors in this cell type; therefore, we can assume that the effects occurring with the addition of melatonin were not mediated by modulation of this hormone on oestrogen receptors. The decreases in cell growth were attributed to a moderate, but significant antiproliferative action of melatonin on this non-hormone-dependent cell line.

Short-term hypothyroidism has no effect on serum leptin concentrations.

The aim of the study is to determine the effect of short-term hypothyroidism on serum leptin levels. For this purpose 30 patients with past medical history of thyroidectomy for differentiated thyroid carcinoma were included. Serum leptin concentrations were similar when patients were on thyrotrophin-suppressive thyroxine therapy than when were admitted 4 weeks after stopping thyroxine treatment to perform a routine 131I scan in hypothyroid status (17.0 +/- s.e.m. 2.14 vs. 17.6 +/- s.e.m. 2.41 ng/ml; p = n.s.). Moreover, no differences were obtained when the analysis was performed separately in men and in women. We conclude that short-term hypothyroidism does not alter serum leptin concentrations. Furthermore, our results suggest that thyroid hormones do not operate through changes in serum leptin levels to regulate energy expenditure.

Genetic, phenotypic and clinical features of acute lymphoblastic leukemias expressing myeloperoxidase mRNA detected by RT-PCR.

Myeloperoxidase (MPO) is found in the azurophilic granules of normal myelocytic cells. Cytochemical staining for MPO activity is used clinically to distinguish myeloid from acute lymphoid leukemias (ALL). However, using a highly sensitive RT-PCR technique, it is possible to detect MPO mRNA in otherwise clear ALL. The significance of this finding remains poorly understood. We have extended our observations to a series of 57 patients with the primary diagnosis of ALL (46 patients tested at diagnosis and 11 cases at relapse). We identified 25 cases (43.8%) of MPO mRNA(+)/enzyme(-) ALL (17 B cell and eight T cell lineage). Expression of myeloid antigens (CD13 or CD33) were detected in nine of them, and remarkably, 18 cases (72%) displayed CD34. Of these 25 MPO mRNA(+) leukemias, 10 (40%) are Bcr-Abl positive (with P210 fusion transcript in five patients while the five remaining cases carried P190 transcript). Moreover, 11 of 16 myeloid negative cases were also negative for any type of Bcr-Abl and MLL rearrangement, indicating that MPO mRNA positivity is not either invariably related to that chromosomal abnormality or necessarily associated with the presence of other myeloid differentiation features. Interestingly, six of these 11 cases are T-ALL, suggesting the presence of some overlapping phase for T and myeloid lineage commitment. Taken together, these findings could suggest a separate biological disease with immature origin and bipotential differentiation capability, which involves B and T-ALL subtypes and should lead to new investigations regarding their prognostic impact.

Penetration of anti-DNA antibodies into immature live cells.

Several data suggest that immature lymphoid cells are more prone to penetration and therefore are affected more by antibodies than their mature counterparts. In this study, we examined the penetration of monoclonal anti-DNA antibodies in several models of immature cells. Results confirm that most anti-DNA antibodies penetrate larger proportions of immature cells than normal adult cells. It was also proven that anti-DNA antibodies induce larger fractions of immature cells to undergo apoptosis than mature cells; however, there is not a numerical association between penetration and apoptosis. Additionally, the penetration and induction of apoptosis of several anti-DNA monoclonal antibodies into U937 and NIH-3T3 cells followed a rather heterogeneous pattern. When mature and immature cells were stimulated polyclonally, it was shown that polyreactive antibodies might act as an accessory signal to induce apoptosis in immature cells. This process could contribute to the edition of the immune repertoire. We propose that naturally occurring polyreactive antinuclear antibodies, through penetration and deletion of self-reacting cells, could participate, either as a unique or secondary signal, in the mechanism of self tolerance. If these polyreactive antibodies undergo affinity maturation, it is possible they may develop into pathogenic antibodies.

Plasma corticotropin-releasing factor in depressive disorders.

BACKGROUND: The aim of this work was to investigate alterations of plasma corticotropin-releasing factor (CRF) levels in depressive states. We have also measured plasma cortisol and corticotropin (ACTH) concentrations and examined their correlation with the peripheral CRF values. METHODS: Thirty-six outpatients from the psychiatric department of a Barcelona hospital who were diagnosed as having major depressive disorder (n = 26) and dysthymic depressive disorder (n = 10) were studied. Among the major depressed patients, 10 suffered from severe depressive disorder and 16 from mild or moderate depressive disorder. The comparison group consisted of 17 healthy volunteers. Cortisol, ACTH, and CRF concentrations were determined by iodine-125 radioimmunoassay; CRF measurements were performed on C18 extracted samples. RESULTS: CRF and cortisol plasma concentrations were significantly higher in major depression and dysthymia than in the comparison group. The major depressed patients did not show significantly different CRF and cortisol levels than the dysthymic. Severe major depressive disorder exhibited significantly higher CRF plasma levels than the mild or moderate episodes. Plasma cortisol and CRF concentrations correlated significantly. CONCLUSIONS: The results obtained indicate that plasma CRF values are altered in depressive disorders and suggest that these determinations could be important for understanding the pathophysiology in affective illness.

Decreased cholecystokinin levels in cerebrospinal fluid of patients with adult chronic hydrocephalus syndrome.

Cholecystokinin (CCK) levels were measured in cerebrospinal fluid (CSF) of patients with adult chronic hydrocephalus syndrome (ACHS) (n = 16) and compared with levels from a control group (n = 11). The CSF concentration of CCK in the ACHS group (0.79 +/- 0.53 fmol/mL) was significantly reduced (p = .002) with respect to the controls (1.55 +/- 0.54 fmol/mL). As CCK-8, the most prevalent from of CCK in the central nervous system, has been demonstrated to play a significant role in several physiological and behavioral actions, the reduced octapeptide values found in ACHS could be involved in the disturbances associated with this disorder. Continuous monitoring of intracranial pressure (ICP) demonstrated different ICP profiles in ACHS. We found that all patients with abnormal ICP records except one showed CCK values under the detection limit. Three of the 4 patients with normal ICP had CCK levels within the normal range. These preliminary studies could evidence that ICP alterations are responsible for part of the loss of brain neuropeptide levels in ACHS.

Evaluation of immune markers in asymptomatic AIDS patients receiving fish oil supplementation.

The effects of oral fish oil (FO) supplementation (8 g/day, capsules) on nutritional status and selected immune markers (CD4/CD8 ratio, IL-1beta, erythrocyte MDA release, dehydroepiandrosterone sulfate [DHEA-S]) were studied in a homogeneous group of asymptomatic HIV-infected patients during 6 weeks. All subjects were classified clinically as A2 according to the CDC revised criteria (mean CD4 count 290 +/-123 cells/mm(3)) and were receiving zidovudine retroviral treatment. The calculated mean energy intake was 3437 +/- 372 Kcal/d, composed of 14% protein, 38% lipids and 48% carbohydrates, and was not modified during the study. The anthropometric parameters, and hematological and plasma biochemistry data showed non-significant changes after FO supplementation. Mean malonyldialdehyde (MDA) release before treatment was: unstimulated 71.5 +/- 37 and stimulated 350.9 +/- 79.8 nmol/g Hb. After FO supplementation (T(6)) MDA release showed unstimulated values of 96.1 +/- 62, and a significant increase after stimulation of 614.1 106.4 nmol/g Hb, which was, however, within the normal range. In the patient's samples, IL-1beta levels in the unstimulated blood culture showed a statistical increase with respect to the normal range before (T(0)) and after (T(6)) FO supplementation with a slight decrease after (mean 49.8 vs 40.9 pg/ml). The stimulated IL-1beta levels after treatment showed a statistically significant decrease that was maintained within the normal range (T(0): 797.7 vs T(6): 535.6 pg/ml). Taken collectively, these results suggest a tendency toward improvement in immune function.

[Angiocentric immunoproliferative lesions: the terminology and the clinical picture]. / Lesiones angiocéntricas inmunoproliferativas: terminología y cuadros clínicos.

Two diagnosed clinical cases of T-cell angiocentric lymphoma are presented, different in their clinical aspects, evolution and treatment. Both represent different spectrums within the entity of angiocentric immunoproliferative lesions. The medical literature is reviewed and emphasis is placed on the different illness which include a wide range from less to more highly malignant within the same entity.

Neurokinin concentrations in cerebrospinal fluid. A preliminary study in Parkinson's disease.

Immunoreactive neurokinin A was measured in the cerebrospinal fluid of twelve patients with Parkinson's disease and eleven normal subjects, using a sensitive and precise extraction/concentration radioimmunoassay method. The mean value obtained in Parkinson's disease patients (13.2 +/- 4.6 pmol/l) was lower than that of the controls (17.4 +/- 5.9). The tendency toward a significant decrease (p = 0.085) found in this preliminary study could indicate that neurokinin A containing neurons are involved in the pathophysiology of Parkinson's disease. In addition, the establishment of reference values for neurokinin A in cerebrospinal fluid may provide a basis for further studies of this neuropeptide in neurological disorders.

Somatostatin cerebrospinal fluid levels in dementia.

Somatostatin levels were measured in cerebrospinal fluid of patients with Alzheimer's disease, multi-infarct dementia and normal pressure hydrocephalus and compared with levels from a normal control group. All pathological groups showed a statistically significant decrease of somatostatin with respect to the control group, but no significant differences were found amongst them. A negative correlation was found between the Mini Mental State Test and the somatostatin levels in Alzheimer's disease patients but not in the other groups. Our results confirm that the lower levels of somatostatin in cerebrospinal fluid are not specific to Alzheimer's disease and indicate that the decrease found in all the groups is probably the result of neuronal destruction or damage in the diseases examined.

Salivary cortisol levels and their correlation with plasma ACTH levels in depressed patients before and after the DST.

OBJECTIVE: The aim of this work was to study the clinical utility of salivary cortisol concentrations in a group of depressed patients undergoing the dexamethasone suppression test (DST) and the correlation of these concentrations with plasma ACTH levels. METHOD: Twenty outpatients from the psychiatric department of a Barcelona hospital who were diagnosed as having nonendogenous (N = 9) or endogenous (N = 11) depression according to DSM-III criteria and the Newcastle scale participated in the study. The comparison group consisted of 12 healthy volunteers. Blood and saliva samples were taken before and after administration of 1 mg of dexamethasone Salivary cortisol and plasma ACTH concentrations were determined by direct iodine-125 radioimmunoassay with commercial kit reagents. RESULTS: Predexamethasone salivary cortisol concentrations were significantly higher in the group with endogenous depression than in the comparison group. A significant correlation was obtained between plasma ACTH and predexamethasone salivary cortisol levels in the group with nonendogenous depression and in the comparison subjects. CONCLUSIONS: These preliminary findings indicate that salivary cortisol could substitute for plasma cortisol in clinical studies in which the DST and hypercortisolemia are evaluated. The lack of correlation between ACTH and cortisol levels in saliva in the group of endogenously depressed patients could indicate a disturbance in the regulation of cortisol secretion in major depression.

Cerebrospinal fluid neuropeptide Y in Alzheimer's disease.

The cerebrospinal fluid neuropeptide Y level was measured by radioimmunoassay in 20 patients with probable Alzheimer's disease and in 19 controls. The mean level was lower in patients (69.5 +/- 36.7 pg/ml) than in controls (103 +/- 21.8 pg/ml; p less than 0.001). Patients with a disease duration of greater than 2 years had cerebrospinal fluid neuropeptide Y levels lower than those with shorter disease duration (p less than 0.02). These results suggest that neuropeptide Y containing cells may be involved in Alzheimer's disease. No correlation was found between neuropeptide Y levels and degree of cognitive impairment or age at disease onset.

Correlation between plasma estradiol and estrone-3-glucuronide in urine during the monitoring of ovarian induction therapy.

An improved radioimmunoassay was developed to determine estrone-3-glucuronide in daily urine. Resulting levels were compared with those of estradiol in plasma of 10 healthy women and 14 undergoing ovulation induction therapy with human menopausal gonadotropin and human chorionic gonadotropin. A highly significant correlation between plasma estradiol and urinary estrone-3-glucuronide in normal (r = .9209; P less than .01) and stimulated (r = .9229; P less than .01) women was demonstrated. These results proved that the pattern of excretion of estrone-3-glucuronide perfectly reflected the changes in plasmatic estradiol levels when monitoring ovarian induction and that estrone-3-glucuronide determinations can provide clinically useful information in human induction therapy.

Plasma levels of beta-endorphin, cortisol, prolactin and growth hormone in depressed patients.

Basal serum cortisol, growth hormone, prolactin and immunoreactive (IR) plasma beta-endorphin levels were measured in 31 depressed patients (14 endogenous, 17 nonendogenous) undergoing the dexamethasone suppression test. The endogenously depressed patients had significantly higher (22.55 +/- 1.34 micrograms/dl) predexamethasone cortisol levels than the nonendogenous patients (16.34 +/- 1.93 micrograms/dl). The mean serum prolactin and growth hormone values of these two groups were not significantly different, while plasma IR-beta-endorphin levels of the endogenous group (40.11 +/- 3.57 pg/ml) were significantly lower than those of the nonendogenous group (120.33 +/- 27.98 pg/ml). Neither group showed a significant correlation between plasma IR-beta-endorphin and serum cortisol values. These results indicate that measurement of predexamethasone serum cortisol values and plasma IR-beta-endorphin could be valuable laboratory tests in the diagnosis of depression.