Pneumococcal whole-cell vaccine: optimization of cell growth of unencapsulated Streptococcus pneumoniae in bioreactor using animal-free medium.

The high cost of the available pneumococcal conjugated vaccines has been an obstacle in implementing vaccination programs for children in developing countries. As an alternative, Malley et al. proposed a vaccine consisting of inactivated whole-cells of unencapsulated S. pneumoniae, which provides serotype-independent protection and involves lower production costs. Although the pneumococcus has been extensively studied, little research has focused on its large-scale culture, thus implying a lack of knowledge of process parameters, which in turn are essential for its successful industrial production. The strain Rx1Al- eryR was originally cultured in Todd-Hewitt medium (THY), which is normally used for pneumococcus isolation, but is unsuitable for human vaccine preparations. The purposes of this study were to compare the strains Rx1Al- eryR and kanR, develop a new medium, and generate new data parameters for scaling-up the process. In static flasks, cell densities were higher for eryR than kanR. In contrast, the optical density (OD) of the former decreased immediately after reaching the stationary phase, and the OD of the latter remained stable. The strain Rx1Al- kanR was cultivated in bioreactors with medium based on either acid-hydrolyzed casein (AHC) or enzymatically hydrolyzed soybean meal (EHS). Biomass production in EHS was 2.5 times higher than in AHC, and about ten times higher than in THY. The process developed for growing the strain Rx1Al- kanR in pH-controlled bioreactors was shown to be satisfactory to this fastidious bacterium. The new culture conditions using this animal-free medium may allow the production of the pneumococcal whole-cell vaccine.

Detección de marcadores de malignidad en nódulos tiroideos por transcripción reversa y reacción en cadena de la polimerasa (RT-PCR) / Detection of malignancy markers in thyroid nodules by reverse transcriptase polymerase chain reaction (RT-PCR)

BACKGROUND: Nodular thyroid disease is a very common disorder with a low frequency of malignancy. The most accurate diagnostic test is fine needle aspiration biopsy (FNAB) of nodules with cytological analysis of the sample. However, this procedure has some limitations in the diagnosis of follicular and papillary thyroid carcinoma. AIM: To detect mRNA from specific malignancy markers in thyroid nodules and to evaluate their potential correlation with cytological and pathological diagnosis. PATIENTS AND METHODS: In 20 patients with thyroid nodules FNAB was performed prior to surgery. The main part of the FNAB sample was used to perform classical cytology. In the remaining of the sample were detected MUC-1, CD26, galectin-3 and TSH receptor mRNAs by RT-PCR technique. RESULTS: Eight patients had positive cytology for papillary cancer, which was confirmed by pathology. Nine had suspicious or non conclusive cytological findings and 3 were negative for neoplastic cells; all 12 were pathologically benign. We detected TSH receptor and galectin-3 mRNA in almost all benign and malignant nodules. MUC-1 was present in 5/8 papillary carcinoma (62.5%), and 1/12 benign nodules (8.3%). CD26 was detected in 7/8 papillary carcinomas but also in 8/12 benign nodules. CONCLUSIONS: RT-PCR can be performed in very small samples of thyroid tissue to detect several mRNA markers. MUC-1 can be a potentially useful marker of malignancy in thyroid nodules. It can be detected by RT-PCR as a complementary technique in the diagnostic evaluation of thyroid nodules.

[Treatment of hyperthyroidism with radioiodine: effects of administered dose on complications and thyroid function]. / Tratamiento del hipertiroidismo con radioyodo: efecto de la dosis administrada sobre la función tiroidea y complicaciones asociadas.

BACKGROUND: Radio-iodine is a definite therapy for Graves disease hyperthyroidism. However, the optimal dosage is still debatable. AIM: To assess the effects of different radioiodine doses on thyroid function and complications in patients with hyperthyroidism. MATERIAL AND METHODS: A retrospective analysis of 139 patients with hyperthyroidism, treated with ratio-iodine between 1988 and 1998. Radio iodine dose used was classified as low (< 10 mCi), intermediate (10-14.9 mCi) or high (> or = 15 mCi). RESULTS: Thirty-five patients were treated with low doses, 33 with intermediate doses and 71 with high doses. There were no differences between these patients in age, disease severity, frequency of post treatment euthyroidism or complications. Patients treated with low doses had a higher frequency of persistent hyperthyroidism than patients treated with high doses (25.7 and 4.2% respectively, p < 0.001). Likewise, the frequency of subsequent hypothyroidism was 60% in patients treated with low doses and 84.5% of those with high doses, in whom it also appeared earlier. Associated complications were clinically irrelevant. In seven patients, Graves ophthalmopathy progressed after treatment, but this progression was not associated with the dose used. CONCLUSIONS: Radio iodine in high doses is useful, safe and effective for the treatment of Graves hyperthyroidism.

[Thyroid cancer. Report of 85 cases]. / Cáncer tiroideo. Comunicación de 85 casos.

BACKGROUND: Thyroid cancer is the most frequent endocrine cancer with important implications in terms of diagnosis and treatment. AIM: To report a population of patients with thyroid cancer diagnosed by pathological studies of the surgical piece. PATIENTS AND METHODS: Eighty five patients (68 female) with the definitive diagnosis of thyroid cancer were studied. Clinical, imaginological, cytological and pathological findings were analyzed. RESULTS: The age range of patients was 10 to 77 years old. Sixty nine patients had ultrasonographic studies which showed a solid nodule in 84%, mixed solid-cystic area in 14.5% and a purely cystic nodule in 1.5% of the cases. Nineteen patients had non specific calcifications. Fine needle aspiration cytology was negative for malignancy in eight patients (false negative rate of 9.9%). The average size of the nodules was of 2.8 +/- 1.6 cm). Six nodules measured less than one cm (microcarcinoma). In the initial surgical procedure, 13 patients had lymph node metastases, 2 of them had a primary tumor of 1 cm and 5 patients had Graves's Disease. Frozen biopsies during operation had 9 false negative results for cancer (10.6%). Pathology showed 64 cases of papillary cancer (75%), 14 of follicular (16.5%), two were Hurthle cell cancer (2.4%), three were medullary (3.5%), and two anaplastic (2.4%). CONCLUSIONS: In our experience, thyroid cancer is more common in women, solid lesions predominate in the ultrasonography and calcifications are frequently found. The tumor size is variable and the most frequent pathological type corresponds to differentiated cancers. Using the definitive pathological study as the standard, the diagnostic sensitivity of fine needle cytology was 90.1%, and of frozen section 89.4%.

Circulating iodide concentrations during and after pregnancy.

Early, indirect studies suggested that an important aspect of thyroid economy during pregnancy was a decline in plasma or serum inorganic iodide (PII) concentrations, but there is little information concerning circulating iodide concentrations as assessed by direct measurement. The present study was undertaken to determine the relationship between gestation and serum iodide concentrations as assessed by direct measurement of PII. PII concentrations, urinary iodide levels, and other parameters of thyroid economy were measured during the first, second, and third trimesters and after delivery in 16 women. Mean serum T4 concentrations were significantly higher in all 3 trimesters than those after delivery. Serum free T4 index concentrations were significantly higher in the first trimester than during later periods of gestation or after delivery, but serum TSH concentrations were not depressed in the first trimester. Serum thyroglobulin concentrations were similar during pregnancy and after delivery. There was wide variability in PII and urinary iodide concentrations during and after pregnancy, but there was no trend for PII concentrations to be depressed during pregnancy. Pregnancy, at least in iodine-sufficient regions, does not have an important influence on circulating concentrations of iodide.

[Influence of maternal feeding restrictions on the mother-daughter dyad]. / Influencia de la restricción alimentaria sobre la díada madre-hija.

Alimentary restraint, cognitive variable related to eating behavior and obesity, is reportedly a valuable predictor for the development of therapeutic strategies. This paper addresses the relationship between maternal restraint and several psychological variables in their daughters (alexithymia, neuroticism, extraversion). From the study of 35 mother-daughter dyads it can be concluded that daughters of highly restrained mothers tend to present high scores in the Restraint scale of the Three Factor Eating Questionnaire of Stunkard and Messick, translated into Spanish and validated as Cuestionario de Conducta Alimentaria. Daughters of highle restrained mothers present also higher scores in the Neuroticism scale of the revised version of the Eysenck Personality Questionnaire. Daughters of mothers with low Restraint scores are in average higher than those of their mothers, although lower than those belonging to daughters of highle restrained mothers. Previous observations on the positive correlation between Disinhibition and Hunger of the Three Factor Eating Questionnaire are confirmed. These results add an additional risk factor for obesity (mothers with high Restraint) and contribute to delineate a set of psychometric indicators which might be useful in the diagnosis and prognosis of eating and body weight disorders.

[Medical treatment of obesity from a multi professional perspective]. / El tratamiento médico de la obesidad desde una perspectiva multidisciplinaria.

Obesity is a serious condition affecting the physical and psychological well being of a large number of individuals. There are many therapeutic options for the disease and their judicious selection is critical. The best results are obtained with well-organised multi professional programs. The aims of the treatment must be defined balancing risks and benefits. Eating behaviour patterns must be considered for the proper selection of therapy. Diet is an essential therapeutic component and has to be individually prescribed for each patient. Pharmacological agents inhibit appetite, increase thermogenesis or inhibit gastrointestinal absorption of nutrients. As these medications have important side effects and complications, extreme care must be taken for their prescription.

[Testosterone-producing adrenal carcinoma: report of 2 cases]. / Carcinoma suprarrenal productor de testosterona: 2 casos clínicos.

We report two women aged 48 and 65 years old with hirsutism and a painful abdominal mass. Laboratory assessment showed serum testosterone levels of 500 and 321 ng/dl (normal values below 110 ng/ml). Abdominal CAT scans showed an adrenal mass in both patients. The younger woman was subjected to a surgical resection, the tumor relapsed two years later and the patient died. The other woman died during the first hospitalization. Adrenal tumors infrequently secrete testosterone as described in these two patients.

Purification of basic fibroblast growth factor and alkaline phosphatase from human placenta.

Human placenta is an available hospital waste which is known to contain many valuable biochemicals that may be commercially exploited. Using placental tissue previously extracted for haemoderivatives, we purified basic fibroblast growth factor (bFGF), a soluble protein, and placental alkaline phosphatase (PALP), a membrane-linked protein, as a coupled process. bFGF purification comprises three steps: extraction and chromatographies on S-Sepharose and heparin-Sepharose. The final product includes a major 17 kDa and a minor 16 kDa component with a specific activity of 8.0 x 10(6) units/mg yielding 0.5-1.0 microgram/kg of placenta. PALP purification comprises four steps: acidic butan-1-ol extraction and chromatographies on Q-Sepharose, concanavalin A-Sepharose and Q-Sepharose. The purified PALP has a molecular mass of 70 kDa, a specific activity of 800 units/mg and yielded 50 micrograms/kg of placenta. The results show the possibility of purifying substances in placental haemolysed blood, soluble products from placental cellular mass and proteins from the cellular membrane in a one-stream process.

[Food restraint and acute stress in women]. / Restriccion alimentaria y estres agudo en mujeres.

According to restraint theory, people scoring high in this dimension of eating behavior should exhibit a conflict between biological set point and cultural norm. In order to explore the hypothesis that this tension should lead to heightened vulnerability to stress, 17 healthy women between 18 and 54 years of age, previously studied from the point of view of eating behavior, were submitted to a five-minute stressful challenge (public speaking). Visual analogue anxiety scales and plasma free fatty acids indicated that high restraint scores predict heightened reactivity to this stressor in women. Such result helps to differentiate further psychological factors related to disorders of eating.

[Insulinoma and pregnancy. Clinical case]. / Insulinoma y embarazo. Caso clinico.

A 25 year old woman developed severe episodes of hypoglycemia during the first weeks of pregnancy. Insulin blood levels of 53 uU/ml and Peptide C levels of 6.5 ng/dl were shown associated to a blood glucose level of 34 mg/dl. Echotomographic examination of the abdomen was unrewarding. Surgical exploration revealed a 1.5 cm tumor located at the junction of the body and tail of the pancreas, which was removed. Hypoglycemia disappeared and pregnancy continued uneventfully ending in a normal delivery at 40 weeks. The newborn was normal and no motor or psychologic abnormalities have been noted during follow up to 2 years of age.

[Patients with Parkinson disease: functional analysis of dopamine receptors by bromocriptine- thyrotropin-releasing hormone test]. / Pacientes con enfermedad de Parkinson: analisis functional de los receptores dopaminérgicos mediante la prueba de hormona liberadora de tirotrofina-bromocriptina.

The bromocriptine-TRH test was performed in 11 patients with advanced Parkinson's disease and in 10 control subjects. Seven patients were receiving treatment with Levo-DOPA, which was discontinued 12 h prior to the test. Basal prolactin levels were 5.88 ng/ml in treated and 18.22 ng/ml in untreated patients as compared to 9.68 mg/ml in controls. Bromocriptine induced a similar reduction of about 60% in prolactin levels in all groups (3.28, 11.5 and 5.95 ng/ml, respectively). After TRH, prolactin levels increased to 13.8 ng/ml in controls and 15.2 ng/ml in patients treated with levo-DOPA. Untreated patients remained with prolactin below basal levels (11.6 ng/ml). We postulate that low basal levels of prolactin in patients treated with levo-DOPA reveal a residual suppressing effect of the drug. The sharp increase after TRH is related to a significant dopaminergic hypofunction as evidenced by clinical findings. A high sensitivity of dopaminergic receptors in the anterior hypofisis is suggested as an explanation for the findings of the test in the untreated patients.

TRH pharmacokinetics and nerve stimulation evoked prostatic fluid secretion during TRH infusion in the dog.

The pharmacokinetics of TRH have been determined in man and the rat as well as other species whose serum contains TRH degrading enzymes, TRH pharmacokinetics have not been determined in the dog. This species is unusual in that its serum contains little or no TRH degrading activity. TRH pharmacokinetics were determined from measurements of plasma TRH concentrations during and following a 1-h infusion of TRH, 20 micrograms.kg-1.min-1, into dogs bearing prostatic urethral cannula. During TRH administration prostatic fluid secretion was evoked by hypogastric nerve stimulation and the content of TRH immunoreactive material in prostatic fluid was determined. Prostatic fluid was also collected after hypogastric nerve stimulation in dogs that did not receive TRH infusion. The TRH plasma profile in TRH-infused dogs exhibited two-compartment characteristics with an initial half-time of 10.4 +/- 5.4 (mean +/- SD) min, a terminal half-time of 69.4 +/- 24 min, and a metabolic clearance rate of 7.64 +/- 4.48 ml.min-1.kg-1. These findings suggest that the half-time of TRH is longer, and its metabolic clearance rate is less, in the dog than in humans or rats. TRH administration was not associated with altered nerve induced prostatic fluid secretion. Prostatic fluid samples contained TRH immunoreactive material regardless of whether they were collected after epithelial cell stimulation with pilocarpine, or after hypogastric nerve stimulation. In contrast to previous results with vasoactive intestinal peptide, TRH does not acutely alter prostatic fluid secretion. As has been reported for prostatic tissue extracts, immunoreactive material is present in prostatic fluid and appears to arise from epithelial cells.

Euthyroid hyperthyroxinemia and thyroxine-binding prealbumin excess in islet cell carcinoma.

This manuscript describes euthyroid hyperthyroxinemia secondary to elevated serum T4-binding prealbumin (TBPA) concentrations in a patient with islet cell carcinoma and reports serum TBPA measurements in other patients with islet cell carcinoma. A 73-yr-old man with a 17-yr history of metastatic islet cell carcinoma was found to have hyperthyroxinemia. His total serum T4 concentration was 18.5 micrograms/dl (normal, 5.5-11.5). Eight years previously, his serum T4 concentration was normal. His free T4 concentration, as determined by equilibrium dialysis, was 1.3 ng/dl (normal, 0.9-2.1). Serum T3, TSH, and T4-binding globulin (TBG) concentrations were normal, as was the TSH response to TRH administration. Polyacrylamide gel electrophoresis of the patient's serum in the presence of tracer amounts of [125I]T4 revealed that, compared to normal sera, [125I]T4 binding to TBPA was increased from 30.0 +/- 6.0% (mean +/- SD) to 52.0%. The distribution of [125I]T3 amont albumin, TBG, and TBPA was normal in this electrophoretic procedure. The concentration of TBPA in the patient's serum was 189 mg/dl. In contrast, the mean serum TBPA concentration in normal men was 40 +/- 4 (mean +/- SD) mg/dl, and that in normal women was 18 +/- 4 mg/dl. Sera from the patient's daughter, his brother, 2 sisters, and 2 male paternal cousins contained normal amounts of TBPA, ranging from 34-47 mg/dl in his male relatives and from 26-30 mg/dl in his female relatives. Serum free T4 index determinations in his relatives also were normal. Serum TBPA concentrations were determined in 14 additional patients with islet cell carcinoma. In 1 of these patients, a man with an insulinoma, serum TBPA was elevated (66 mg/dl). This patient's serum T4 level was 7.3 micrograms/dl, and his free T4 index was 7.0. These data and another study suggest that islet cell carcinoma may rarely produce a TBPA-like protein resulting in an elevated serum TBPA concentration. Markedly elevated serum TBPA is associated with euthyroid hyperthyroxinemia.

Adrenal rest tumor of the liver causing Cushing's syndrome: treatment with ketoconazole preceding an apparent surgical cure.

Ketoconazole, an imidazole derivative known to inhibit cytochrome P450-dependent adrenal enzymes was given to a patient with a functioning adrenal rest tumor of the liver in preparation for surgery. The drug was administered in a stepwise manner for 42 days starting with 400 mg and reaching 1 g the last 4 weeks of the trial. Clear clinical improvement was evident early in the trial and was associated with evidence of amelioration of her hypercortisolism and striking changes in serum and urinary levels of steroid hormones and metabolites. Sex steroids in serum and urine fell dramatically from the first day to the end of the trial. Urinary 17-ketosteroid excretion fell from a basal average of 139 mg/24 h to near normal levels within a week of therapy; serum testosterone fell from a basal level of 2.4 to 0.18 ng/ml; serum 17 beta-estradiol fell likewise from 1096 to 150 pg/ml. In contrast, cortisol levels in serum and urine increased in the first 2 weeks of the trial and subsequently fell to values below the basal levels. Similarly, serum 17 alpha-OH-progesterone levels increased 63% above the basal levels by day 6 of the trial and declined afterwards. Nine months after successful tumor resection the patient is apparently cured as judged by steroid hormone levels and physical appearance. We conclude that ketoconazole was effective in blocking tumoral steroidogenesis which resulted in clinical benefit.