Differentiation of green, white, black, Oolong, and Pu-erh teas according to their free amino acids content.

In this paper, the differentiation of green, black, Oolong, white, and Pu-erh teas has been carried out according to their free amino acid contents. Alanine, arginine, asparagine, aspartic acid, glutamic acid, isoleucine, histidine, leucine, phenylalanine, serine, theanine, threonine, and tyrosine have been determined by liquid chromatography with derivatization with o-phthalaldehyde and fluorescence detection. The chromatographic separation was achieved with a Hypersil ODS column and gradient elution. The amino acid contents were used as chemometric descriptors for classification purposes of different tea varieties. Principal component analysis, k-nearest neighbors, linear discriminant analysis, and artificial neural networks were applied to differentiate tea varieties. Using back-propagation multilayer perceptron artificial neural networks, 100% success in the classification was obtained. The most differentiating amino acids were glutamic acid, asparagine, serine, alanine, leucine, and isoleucine.

Neuropeptides bombesin and calcitonin induce resistance to etoposide induced apoptosis in prostate cancer cell lines.

BACKGROUND: Neuroendocrine differentiation in prostatic carcinoma has been related to regulation of proliferation and metastatic potential and correlated with prognosis. More than 80% of prostate carcinomas initially respond to androgen ablation, but most relapse, due to the heterogeneous presence of androgen-dependent and independent clones. The pathways of cellular proliferation and apoptosis are inexorably linked to minimize the occurrence of neoplasia, and disfunction of apoptosis is proposed as a pathogenic process in malignant tumors. Androgen-dependent prostatic cancer cells undergo apoptosis after androgen deprivation, but not androgen-independent ones due to a defect in the initiation step. Anyway, they retain the basic cellular machinery to undergo apoptosis. We suggest a possible role of neuroendocrine differentiation in the onset and regulation of apoptosis in prostatic neoplasia. METHODS: LNCaP, PC-3 and DU 145 prostatic cancer cell lines were induced to undergo apoptosis after treatment with etoposide alone or plus androgen ablation. We tested the role of neuropeptides bombesin and calcitonin at modulating etoposide induced apoptosis. RESULTS: Etoposide-induced apoptosis in all cancer cell lines was achieved. In LNCaP androgen ablation was also required. Apoptosis is prevented in all three lines when bombesin was added. Calcitonin addition prevents apoptosis in PC-3, LNCaP and in an etoposide dose-dependent way in DU 145. CONCLUSION: Neuropeptides bombesin and calcitonin can modulate the apoptotic response of prostate cancer cells by inducing resistance to etoposide-induced apoptosis, suggesting that neuropeptides can be used as a target of therapeutical approach in prostatic carcinoma.

AgNOR and breast cancer. A study by image analysis.

The nucleolar organizer regions are used as a new marker in proliferation cell. AgNOR are loops of DNA actively transcribing to ribosomal RNA and they are associated with nonhistone proteins with strong argyrophil. In the present study we correlated the relationships between number and size of AgNOR measured by an automatic image analysis using CAS-100, in eleven breast carcinomas in different clinical stages and histological grades. We could identify the AgNOR with silver stain; the relationship between the number/size and histological grade/clinical stage shows that in normal breast we find one or two AgNOR but this number is larger when the carcinomas are less differentiated. When studied, the size is larger in the normal breast. We can make the same conclusions with the carcinomas which are lymph-node +.

Neuropeptides, apoptosis and ion changes in prostate cancer. Methods of study and recent developments.

It has been suggested that neuroendocrine (NE) cells provide paracrine stimuli for the propagation of local carcinoma cells and that NE differentiation is associated with the progression of prostate cancer toward an androgen-independent state. Apoptosis comprises a critical intracellular defense mechanism against tumorigenic growth and is associated with a number of changes in the elemental content of the cell. The neuropeptides bombesin and calcitonin, which inhibit etoposide-induced apoptosis, also inhibit the etoposide-induced elemental changes in prostate carcinoma cells. This important fact strengthens the link between apoptosis and changes in the intracellular elemental content. This protective effect on etoposide-induced apoptosis appears to be quite similar in androgen-dependent and androgen-independent cell lines. This confirms that neuropeptides confer antiapoptotic capabilities on non-neuroendocrine cells in close proximity to neuroendocrine cells. It can therefore be speculated that certain neuroendocrine peptides can increase the survival and further growth of neighboring cells and may thereby contribute to the aggressive clinical course of prostate tumors containing neuroendocrine elements. In addition, this correlation provides an objective basis for the study of neuropeptide target points and may be helpful for alternative therapeutic protocols using neuropeptide inhibitors in the treatment of patients with advanced prostatic carcinoma. The culture techniques described were, thus, designed in order to achieve two important goals. First, the development of an in vitro model that allows an approach to neuroendocrine differentiation in prostate cancer and its role in apoptosis blockage. Second, the method has been designed in order to permit rapid cryofixation of intact cell monolayers for subsequent x-ray microanalysis.

Etoposide sensitivity of human prostatic cancer cell lines PC-3, DU 145 and LNCaP.

Metastatic prostatic cancer is typically refractory to androgen ablation therapy due to the presence of androgen-independent clones in the neoplasia. A therapeutical approach which could effectively control androgen-dependent and independent cells is, thus, needed. Maybe the failure of certain cancer cells to engage in apoptosis could explain the inherent drug resistance of many tumors. Anyway, these cells can retain the ability to undergo apoptosis in response to an adequate stimulus. We tested whether etoposide, a topoisomerase II inhibitor, could induce apoptosis in androgen-dependent (LNCaP) as well as independent (PC-3 and DU 145) human prostate cancer cell lines. Morphological examination was performed, as it is regarded as one of the most reliable parameters for the detection of apoptotic changes. Complementarily, biochemical and flow cytometric studies were also used. Characteristical changes of apoptosis were demonstrated in PC-3, Du 145, and LNCaP cancer cells after treatment with etoposide. These cells, thus, retain the ability to undergo apoptosis under adequate conditions, in a promising approach to hormone refractory prostate cancer therapy.

Determination of plasma malondialdehyde-like material and its clinical application in stroke patients.

Plasma malondialdehyde-like material (MDA-LM) was evaluated in 138 normal subjects and in a group of 57 stroke patients using a modification of the method of Smith et al. (1976). The basal level of MDA-LM in the control group was 35 mumol/l with a range of 22-50 mumol/l. Values above 50 mumol/l were found in 80% of the patients suffering from subarachnoid haemorrhage, in 68% of those with cerebral thrombosis, and in 17% with transient ischaemic attacks. None of the patients with cerebral embolism, intracerebral haematoma, or lacunar infarct had values above 50 mumol/l. Significant statistical differences were found between the control group and all the patients except those with lacunar infarcts.

Bioelectrical tissue resistance during various methods of myocardial preservation.

Twenty-one mongrel dogs underwent cardiopulmonary bypass (CPB) and ischemic cardiac arrest for 60 minutes. The bioelectrical tissue impedance, water content, and ultrastructure of the myocardium were studied and correlated with various types of myocardial protection applied. The animals were divided into three groups according to the myocardial protection applied: Group 1, moderate general hypothermia; Group 2, moderate general and local hypothermia; Group 3, moderate general hypothermia and crystalloid cardioplegia. The hearts from 3 normal dogs served as controls. After 35 minutes of ischemic arrest, the difference in bioelectrical impedance of the myocardium of the left ventricle between Groups 1 and 2 was highly significant (p less than 0.001); in Groups 2 and 3, a significant difference was observed after 50 minutes. The heart water content in all three groups increased over that found in the hearts of the control animals, but there was no significant difference between Groups 2 and 3. There was evidence by ultramicroscopic study of ischemic damage to the myocardium in all of the experimental groups. Damage was less evident in Groups 2 and 3. We conclude that measurement of myocardial bioelectrical tissue resistance is a sensitive indicator of myocardial damage due to ischemia and may be useful clinically in determining the need for and type of cardiac resuscitation, particularly following CPB.

Preoperative treatment of ruptured intracranial aneurysms with tranexamic acid and monitoring of fibrinolytic activity.

The fibrinolytic activity in cerebrospinal fluid has been monitored by determination of levels of fibrin split products (FSP) in 23 patients with ruptured intracranial aneurysms. In 20 of these 23, FSP was found in the cerebrospinal fluid (CSF), with levels ranging from 10 to 80 micrograms/ml. Eleven of the 23 patients were treated with 2 gm tranexamic acid daily. In these patients FSP was found in only two cases during the 2nd week, while in 12 untreated patients it was found in 10 cases. These results suggest that there exists a localized fibrinolytic activity, and monitoring the FSP levels in the CSF may be a simple and accurate method for controlling the efficiency of antifibrinolytic therapy. Thus, treatment could be begun with a lower dose, which could be increased later as deemed necessary from the results of careful monitoring.

[Relation of prolactin with nodular hyperplasia and carcinoma of the prostate]. / Relación de la prolactina con la hiperplasia nodular y el carcinoma de próstata.

The relationship between prolactin in NH and prostate carcinoma has several aspects: under normal conditions, prolactin enhances androgens output, increasing the Leydig cell susceptibility to LH. At the prostate level, it increases testosterone binding to the prostate epithelium and stimulates the conversion of testosterone to dihydrotestosterone. On its own, it decreases prostatic protein degradation and increases the DNA synthesis index. With regard to Nodular Hyperplasia, once its hormone dependency is rated, it also appears to be stimulated by high levels of prolactin. The NH cell cultures with human prostate increase their growth, cell division, as well as the DNA synthesis. The aspects of the prolactin-prostate carcinoma interaction involve considering this hormone as a possible carcinogenic agent, emphasizing the existence of high plasma levels of this hormone in individuals with prostate carcinoma, where any androgen excess is considered a major factor at the genesis of this tumour, and prolactin increases indirectly its testicular synthesis. Also, prolactin itself stimulates the growth and development of carcinomatous human cell lines, increasing its effect in the presence of testosterone. For this reason, controlling this hormone levels becomes an element to have into consideration in the treatment and follow-up of these patients; also, the reevaluation of any drug therapy that, besides the initial goal pursued, does not maintain baseline prolactin levels. The association of anti-prolactin drugs has to be understood as a co-adjuvant therapy when there is androgenic deprivation, never as a single treatment.

[Application of transmission and scanning electron microscopy in experimental obstructive uropathy: II. Tubular lesions]. / Aplicación de la microscopia electrónica de transmisión y barrido en la uropatía obstructiva experimental: II.--Lesiónes tubulares.

The morfoestructural changes that appear at renal tubule level in obstructive uropathy consist in a progressive loss of the differentiations in the plasmatic membrane: reduction in the number of apical microvilli and of basal invaginations, which entails a drop in the exchange surface area. Degeneration and cellular necrosis phenomena also exist, with the appearance of cell traces in the tubule lumen. On the other hand, morphological signs of a rapid rate of cellular activity in the tubule cells are displayed. Together with the modifications that take place at basal membrane level these changes give rise to an alteration of tubule dynamics.

[Application of transmission and scanning electron microscopy in experimental obstructive uropathy: III. Lesions at the interstitial level]. / Aplicación de la microscopia electrónica de transmisión y barrido en la uropatía obstructiva experimental: III.--Lesiones a nivel del intersticio.

In experimental obstructive uropathy the renal interstice undergoes profound modifications, from the earliest stages, with the fibroblastic elements adopting a palisade arrangement between the renal tubules and the blood cells. Between these cells there is a marked edema configuring "canals", which could be the morphological image of the tubulovenous reflux described in obstructive uropathy. these routes, which we will call tubule-interstice-vein "canals", since in no case could we verify the existence of a direct communication between the tubule formations and the veins, disappear in animals put down late, where fibro and myofibroblastic elements, round cells and macrophages proliferate. A larger number of fibres and fundamental substance are also found. As a result of these changes, a great alteration in renal architecture takes place.

[Application of transmission and scanning electron microscopy in experimental obstructive uropathy: I. Glomerular lesions]. / Aplicación de la microscopía electrónica de transmisión y barrido en la uropatía obstructiva experimental: I.--Lesiones glomerulares.

In this research we use the Electronic Transmission and Sweep Microscope to study the morphostructural modifications undergone by the renal glomerulus in experimental obstructive uropathy. The most significant alterations take place at the level of the basal lamina of the glomerulus, where progressive thickening anf folding occurs. The podocytes display, mainly in the late groups, loss of the digit-shaped character of their secondary prolongations, with shortening and thickening of these. They also take on an anarchic arrangement and are pseudofused to one another, forming a cellular lamina in contact with the underlying basal membrane. These morphostructural changes justify the reduction in glomerular filtration that takes place in obstructive uropathy.

[Embryonal development of interstitial Leydig cells in the rat testis]. / Desarrollo embrionario de las células intersticiales de Leydig en el testículo de rata.

Blood testosterone concentrations are not constant throughout life, there being a direct correlation with the numbers of cells producing that hormone, for which reason the existence of two Leydig's cells populations has been proposed: one foetal formation, which disappears just prior or immediately after birth, and a subsequent adult generation. In recent studies, however, that withdrawal has not been observed. Considering all the above, we have studied the evolution of these elements during the rat foetal period. From a morphological point of view Leydig's cells do not differentiate from those in the gonadal interstice until day 17 of pregnancy, showing a considerable increase in their numbers by day 21: while from that day onwards their numbers decrease they never disappear completely. We believe those cells become differentiated from the interstitial mesenchymal cells, and that their increase is facilitated by the proliferation of these same elements.

[Post-natal development of interstitial Leydig cells in rat testicle]. / Desarrollo postnatal de las células intersticiales de Leydig en el testículo de rata.

The existence of different stages of postnatal activities in Leydig's cells, as demonstrated in hormone studies, appears to be fully admitted. Such occurrence should have repercussions on the population of those elements primarily entrusted with their production, either in the number of elements or their activities. The present article studies Leydig's cells rate in Wistar rats over the postnatal stage, from birth to adulthood. During this postnatal stage, Leydig's cells population does not actually disappear. There is a decrease immediately after birth and up to the age of 21 days when gonadal drop occurs in this species, and from there on a considerable increase takes place for in a two-month period they multiply by ten. On the other hand, it is highly surprising the great speed of testicle growth up to the age of 60 days, reaching maximum size at 90 days. The increase is due to the growth of the tubular compartment.

[The nuclear roundness factor as a parameter in the anatomoclinical correlation of prostatic carcinoma]. / El factor de redondez nuclear como parámetro en la correlación anatomoclínica del carcinoma prostático.

This paper correlates the nuclear roundness factor to clinical stage, patient's age and acid phosphatase level at the time of diagnosis in a group of 48 patients with prostate carcinoma and a minimal follow-up of 5 years. It has been demonstrated that, from an anatomo-clinical point of view, the nuclear roundness factor is a good indicator because it presents increasing values as the clinical stage of tumoral extension worsens, and in those stages where the number of patients was relatively high the average of deaths was always higher than that of survivors. On the other hand, a direct correlation to total acid phosphatase, and an inverse correlation to age at the time of diagnosis was shown in patients with changes in these values.

[Evaluation of nuclear roundness as a tool for prostatic carcinoma grading]. / Valoración del factor de redondez nuclear como herramienta para la gradación del carcinoma prostático.

The microscopic patterns adopted by prostate carcinomas are extraordinarily variable making its histopathological classification and grading extremely complicated. There are numerous systems for prostatic tumours' grading, but many of them lack a truly reproducible character and an acceptable degree of reliability; for this reason, we have studied a series of nuclear microscopic parameters, from a quantitative point of view, in a series of prostatic carcinomas with a minimum follow-up of five years. It was observed that of the morphometric values obtained--area, perimeter, projection on the X- and Y-axis as well as the nuclear roundness factor--the most useful and representative is the latter. Besides, this is an objective parameter to evaluate nuclear anaplasia and can also be considered as a prognostic factor since, in our series, those who died had a greater mean nuclear roundness factor than those who survived.

[Usefulness of carbohydrate-deficient transferrin (CDT) in the assessment of abuse and abstinence in chronic alcoholism]. / Utilidad de la transferrina deficiente en carbohidratos (CDT) para la valoración del abuso y la abstinencia en el alcoholismo crónico.

OBJECTIVE: To evaluate the utility of carbohydrate deficient transferrin (CDT) for the chronic alcoholism diagnostic, the recent intake and the abstinence, comparing it with the most recognized biological markers, specially the MCV and the GGT. METHODS: We study 87 subjects: 22 controls, 42 subjects abstinent chronic alcoholics and 23 patient active alcoholics. From the last group we subject the middle to a 15 days abstinence period. We compare the CDT with the mos recognized markers in the difference groups. RESULTS: We found significant difference with MCV and GGT in the chronic alcoholism diagnostic. There were significant difference with MCV, GGT and specially CDT in the case of recent alcoholic intake. CDT was the only one that shows significant difference in recent abstinence. CONCLUSION: According to our results the CDT is a poor marker of chronic alcoholism. The classical markers (MCV nd GGT) are better on tha propriety. CDT, however, is a valid and effective marker for the recent alcohol intake, it is better than GGT and MCV. And it is the only valid marker for the recent abstinence. So, we estimate that, it would be advisable the CDT clinical use with restringed criterions for the recent alcoholic intake diagnostics and, specially for the abstinence one.

[Descriptive study of ambulatory patients with alcohol-related liver disease in our setting]. / Estudio descriptivo de los pacientes ambulatorios con enfermedad hepática por alcohol en nuestro medio.

AIMS: We tried to show the demographic characteristic and alcohol intake habits among our outpatients. We study the influence of age, sex, habitat and socioeconomical status on alcoholic habit. DESIGN: Retrospective and institution based study. Patients. 164 patients who were followed up for alcohol liver disease in our outpatient section. RESULTS: Average age to start drinking alcohol was 18.6 (7.36) years, years of alcoholism were 35.4 (13.5) years, average daily alcohol intake was 161.2 (116.7) grams of pure alcohol. Only 16 men (8%) drank less than 60 grams a day. 5 (35.7%) women drank less than 40 grams a day. Life-cumulative alcohol intake was correlated with Maddrey's score at the end of the study (r = +0.407). Average daily alcohol intake was correlated with ultrasonographic features of the liver (r = +0.283), we appreciated that Prothrombin Time was also correlated with ultrasonographic features of the liver (r = +0.301). The percentage of patients who suffer, at least one decompensation of their disease was 39%. CONCLUSIONS: Average age to start drinking is about legal age. Life-cumulative alcohol intake was related to Prothrombin Time and ultrasonographic features of the liver.

[Embryonary rhabdomyosarcoma of the middle ear. Apropos of a case]. / Rabdomiosarcoma embrionario de oído medio. A propósito de un caso.

The subject presented for its rarity is a case of rhabdomyosarcoma embryonal ear in girl patient 6 years old. Her clinic first gave a wrong diagnosis of chronic otitis of the middle ear with otorrhea and facial paralysis. A malignant tumour was suspected during the operation. A histopathology study confirmed the diagnosis of rhabdomyosarcoma. A review of the literature is made and details given of how often it occurs, and incidence, histopathology types, treatment and prognosis.

[X-ray microanalysis of epitheliomas and melanomas: is the Cu/Zn index a prognostic parameter? Preliminary note]. / Microanálisis de rayos X en epiteliomas y melanomas: es el índice Cu/Zn un parámetro pronóstico? Nota previa.

The authors comment the interest of the X Ray microanalysis in Dermatology and, especially, the Cu/Zn index determination in cutaneous tumors.