Evaluation of adequacy of levo-thyroxine dosage in patients with differentiated thyroid carcinoma: correlation between morning and afternoon TSH determination.

PURPOSE: The aim of this study was to judge the reliability of evaluating thyroid-stimulating hormone (TSH) and free thyroxine (f-T4) in the morning and afternoon in differentiated thyroid carcinoma (DTC) patients. METHODS: We evaluated 153 DTC patients, aged 61 ± 13 years, in active follow-up in our center after primary treatments and under stabilized levo-thyroxine (L-T4) posology. In each patient, morning and afternoon examinations were performed 1-3 months apart. Blood samples were collected at 08:00-09:00 h and 15:00-16:00 h. TSH and f-T4 were evaluated in both samples. Thyroglobulin (Tg), Tg-antibodies and neck ultrasonography were also evaluated. RESULTS: According to clinical and laboratory examinations, 92% of patients were disease-free, 6% had biochemical disease, and 2% structural disease. L-T4 dosages (1.64 ± 0.38 µg/kg b.w.) proved the same on both occasions, despite slight changes in body weight or L-T4 posology in 15% of patients. Free-T4 values were significantly higher in the afternoon (21.5 ± 0.3 pmol/L) than in the morning (18.8 ± 0.4 pmol/L; P < 0.0001), whereas TSH values were statistically unchanged (morning 0.85 ± 0.25 mIU/L; afternoon 0.72 ± 0.20 mIU/L). There was a significant correlation (P < 0.0001) between the two TSH determinations in the same patients. CONCLUSIONS: In DTC patients, follow-up examination consists of clinical and laboratory evaluations. The majority of patients have good disease control. Our study suggests that the adequacy of L-T4 therapy can be monitored equally well either in the morning or in the afternoon. Afternoon examinations can alleviate crowding in hospital ambulatories in the morning.

Plasma estrone sulfate concentrations and genetic variation at the CYP19A1 locus in postmenopausal women with early breast cancer treated with letrozole.

Estrogen synthesis suppression induced by aromatase inhibitors in breast cancer (BC) patients may be affected by single nucleotide polymorphisms (SNPs) of the gene encoding aromatase enzyme, CYP19A1. We assessed the association between plasma estrone sulfate (ES), letrozole treatment, and four SNPs of CYP19A1 gene (rs10046 C>T, rs4646 G>T, rs749292 C>T, rs727479 T>G) which seem to be related to circulating estrogen levels. Patients were enrolled into a prospective, Italian multi-center clinical trial (Gruppo Italiano Mammella, GIM-5) testing the association of CYP19A1 SNPs with the efficacy of letrozole adjuvant therapy, in postmenopausal early BC patients. SNPs were identified from peripheral blood cell DNA. Plasma ES concentrations were evaluated by Radio Immuno Assay. Blood samples were obtained immediately before letrozole therapy (N = 204), at 6-weeks (N = 178), 6 (N = 152) and 12-months (N = 136) during treatment. Medians (IQR) of ES were 160 pg/mL (85-274) at baseline, 35 pg/mL (12-64) at 6-weeks, 29 pg/mL (17-48) at 6 months and 25 pg/mL (8-46) after 12 months treatment. No statistically significant association was evident between polymorphisms and ES circulating levels during letrozole therapy. Letrozole suppression of the aromatase enzyme function is not affected by polymorphisms of CYP19A1 gene in postmenopausal BC patients.

Increased serum levels of soluble CD44 standard, but not of variant isoforms v5 and v6, in B cell chronic lymphocytic leukemia.

The CD44 cell surface proteoglycan participates in a variety of functions including lymphohematopoiesis, lymphocyte homing and tumor metastasis. In addition to the standard form (CD44st), a large family of variant isoforms (CD44v) is generated by alternative splicing of a single gene. Certain CD44v (v5 and V6) are upregulated in the course of neoplastic progression and reflect the metastatic potential of tumor cells. CD44 v6 is expressed in high-grade non-Hodgkin's lymphoma cells and is released in the serum, thus providing a soluble marker that reflects tumor burden, disease progression and treatment response. Here we show that serum CD44st is elevated in approximately half of B-CLL patients. In contrast, CD44v5 and v6 are detected at normal levels in the large majority of the cases. CD44st serum levels correlate significantly with the number of circulating leukemic B cells and with the levels of another soluble B-CLL marker, beta2-microglobulin. Immunoprecipitation analyses of B-CLL sera allow detection of several high molecular weight bands and of a 78 kDa band that represents a soluble form of CD44st and is 4 kDa lower than a similar band (82 kDa) detected in B-CLL cell lysates. Elevated serum CD44st associates with a number of unfavorable prognostic factors such as high peripheral blood lymphocytosis, splenomegaly, advanced disease stage and therapy requirement. A follow-up study indicates that serum levels of CD44st are related to disease status, thus reinforcing our veiw that this molecule may represent a reliable tumor marker in B-CLL.

Biochemical characterization and membrane expression of an antigen shared by activated and neoplastic cells of neuroectodermal origin.

The reactivity of a mAb (M16) raised against a small cell lung carcinoma line is described. M16 identifies a surface antigen expressed on cells of neuroectodermal origin following activation, as well as neoplastic transformation. M16 antigen expression is increased on retinoblastoma and neuroblastoma cell lines upon 'in vitro' stimulation and it is induced 'in vivo' on glial cells activated following brain injury. Furthermore, glial tumors show levels of M16 molecule expression increasing with the degree of malignancy, and in a retinoblastoma cell line, the expression of M16 was inversely related to the level of HLA-Class I and N-CAM antigens. The M16 antigen may represent a marker of both activation and neoplastic progression for neuroectodermal cells.

Diagnostic value of CYFRA 21-1 tumor marker and CEA in pleural effusion due to mesothelioma.

STUDY OBJECTIVE: The aim of our study was to assess the clinical value of CYFRA 21-1 tumor marker and carcinoembryonic antigen (CEA) as diagnostic tools that are complementary to cytology in the diagnosis of malignant mesotheliomas. PATIENTS: We measured CEA and CYFRA 21-1 in the pleural effusions (PEs) and serum of 106 patients (benign lung disease, 34 patients; bronchogenic and metastatic carcinoma, 40 patients; mesothelioma, 32 patients). METHODS: CEA and CYFRA 21--1 levels were determined by means of two commercial enzyme immunoassays. RESULTS: The cutoff levels of CYFRA 21--1 and CEA in malignant PEs, selected on the basis of the best diagnostic efficacy, were 41.9 ng/mL and 5.0 ng/mL, respectively. In all neoplastic PEs, CYFRA 21--1 and CEA sensitivity was 78% and 30.6%, respectively, with a specificity of 80% and 91%, respectively. The sensitivity of CYFRA 21--1 and CEA in patients with mesothelioma was 87.5% and 3.1%, respectively. The results of the CYFRA 21--1 assay were positive in 17 of 19 cases of mesothelioma (89.5%) with a negative or uncertain cytology. The association of the tumor marker assay and the cytology allowed a correct diagnosis in 30 of 32 cases of mesothelioma (93.7%). CONCLUSION: This study suggests that CYFRA 21--1 would provide a useful parameter for the differential diagnosis between benign and malignant PE from mesothelioma when the result of cytology is negative or uncertain and the clinical context does not allow a more aggressive approach. Moreover, the association of CYFRA 21--1 with CEA could provide details for a differential diagnosis between mesotheliomas and carcinomas. In fact, an elevated CYFRA 21--1 level with a low CEA level is highly suggestive of mesothelioma, whereas high levels of CEA alone or high levels of both the markers suggest a diagnosis of malignant PE, excluding mesothelioma.

Endocrinological and clinical evaluation of two depot formulations of leuprolide acetate in pre- and perimenopausal breast cancer patients.

PURPOSE: To evaluate the endocrinological and clinical activity of a new slow-release formulation of leuprolide acetate in breast cancer patients. METHODS: A total of 50 pre- or perimenopausal patients with early- or late-stage breast cancer who were candidates for endocrine treatment were included in the study and randomly allocated to receive either 3.75 mg of leuprolide acetate every month or 11.25 mg of leuprolide acetate every 3 months. Patients were treated until disease recurrence or progression or for a maximum of 24 months. Treatment outcome, side effects, and serum levels of gonadotrophins, estradiol, progesterone, and delta4-androstenedione were analyzed at different time points. RESULTS: In all, 23 patients were allocated to the monthly formulation and 27, to the 3-monthly formulation. The median time on treatment was comparable. There was no evidence of any difference in clinical outcome or drug-induced side effects, hot flushes being recorded in about 50% of patients in both groups. Altogether, 35 patients were actively menstruating at the beginning of treatment; all of them became amenorrhoic after 3 months and remained so until treatment with leuprolide was continued, irrespective of the allocated treatment. All endocrine parameters, particularly estradiol levels, were suppressed to a similar extent. CONCLUSIONS: The present results indicate that the two formulations exert a comparable estrogen-suppressive effect and warrant further study of the 3-monthly formulation of leuprolide acetate in breast cancer patients.

Effects of stress on adrenal steroidogenesis blockade by aminoglutethimide.

After one week of AG-treatment, the adrenal cortex reveals a marked storage of lipids and cholesterol, as well as a decrease in haematic corticosterone and progesterone levels. On the contrary, when AG-treatment is associated with stress, an increase in plasma corticosterone and progesterone levels occurs, together with a less evident lipidic storage in the adrenal cortex. These results seem to indicate the possibility that, during stress, hypothalamo-hypophysis axis activation is able to remove, at least partially, the adrenal biosynthesis blockade operated by AG-administration.

A morphological and morphometrical study of displaced amacrine cells in dog retina.

The morphology and morphometry of displaced amacrine cells (DACs) have been investigated in dog retina with the aim of finding out differences in relation to their distance from the optic papilla and to their location within the nasal and temporal retina. The study was performed by using the Boycott and Peichl's silver impregnation method in wholemount retina (12) while the morphometry of the cell body and of the dendritic field was carried out by Leitz ASM 68K image analyser. No morphological difference was found between nasal and temporal cells although a variability, sometimes significative, in their morphometry was observed. Moreover, the morphological features of the DACs remained constant even by varying their distance from the optic papilla.

Neuropeptide Y in the brain and retina of the adult teleost gilthead seabream (Sparus aurata L.).

The presence of neuropeptide Y (NPY) in the brain and retina of gilthead seabream (Sparus aurata L.) was investigated for the first time. For this investigation we employed an immunoperoxidase technique and the western immunoblot analysis using an antiserum raised against porcine NPY. The results showed that NPY-immunoreactivity was widely distributed in the brain of S. aurata. In particular, we have found NPY-immunoreactive (ir) neurons in the area ventralis telencephali pars centralis and pars lateralis, in the area dorsali telencephali pars centralis subdivision two and in nucleus intermedius thalami. An intense NPY-ir was detected in the telencephalon, in the optic tectum, in the thalamus, hypothalamus and in the vagal lobes. Scarce positive fibres were seen in the olfactory bulbs. NPY-ir amacrine cells were observed in the retina. The western immunoblot analysis revealed a protein band with a mobility corresponding to that of synthetic NPY. Our findings are, in general, in agreement with those obtained in other teleosts. The extensive distribution of NPY indicates for this peptide a key role in basic physiological actions, including visual and gustatory inputs processing.

Dog retinal ganglion cells: a morphological and morphometrical study in aging.

Using a reduced-silver method, this study examined ganglioncell morphology and morphometry in aging dog wholemount retina. The ganglion-cell body size and topography were obtained using a semi-automatic image analyser (Leitz ASM 68K, Wetzlar, Germany) studying a sector (with an angle of 10 degrees centred on the optic papilla) as a sampling area from the superior, middle and inferior regions of nasal and temporal retina. The ganglion-cell type and topography of the elder dog retina resembled those of cat retina, with the exception of a lack of alpha cells in the peripheral area of the temporal retina. However, the morphological and morphometrical features are not influenced by aging.

The spinal cord development in guinea pig: a morphometric study on an image analysis system.

Using an image analysis system, the Authors carried out a morphometric study on guinea pig spinal cord in order to determine volumetric changes of white and gray matter during development. White and gray matter volumes were determined by measuring the area occupied by these matters in 10 micrograms sections of spinal cord in 1 day and 90 days old subjects. Several topographic correspondences in the localisation of the lowest and highest volumetric values were observed in the two groups of subjects. Such correspondences were more marked for white than gray matter. Moreover, during growth white matter volume showed an increase double that observed in gray matter.

The neurochemical maturation of the rabbit cerebellum.

The immunocytochemical distribution of several neuronal and glial antigens was investigated in the cerebellum of the developing and adult rabbit. Neurofilament positive neurons appeared at embryonic day (E) 25. Purkinje cells transiently expressed neurofilament polypeptides from postnatal day (P) 0 to 15. At later postnatal ages, staining was localised to the parallel fibres, the axonal arbors of the basket cells and fibres of the white matter. Neuron specific enolase (NSE) immunoreactivity was first detected at E25. At P0 Purkinje cells were positive and their staining intensity increased up to P25. From P30 to adulthood virtually all cells in the molecular and Purkinje cell layers were stained. Scattered PGP 9.5-immunoreactive neurons appeared in the cerebellar anlage at P25. Purkinje and Golgi cells were labelled by P0. Synaptophysin immunoreactivity was first observed at P0 in the form of a fine punctate reaction surrounding the perikarya and proximal dendrites of Purkinje cells. By P10, it became particularly intense within the cerebellar glomeruli of the granular layer. Neurons of the deep cerebellar nuclei expressed NSE and PGP 9.5 starting from E25. GFAP and S-100 immunoreactivities were first detected at P10. GFAP-immunopositive astrocytes progressively increased in number up to adulthood. S-100-immunoreactive glial cells were detected throughout the white and grey matter. Bergmann glial cells and their fibres were strongly immunoreactive. Vimentin positive glial cells and fibres were first observed at E15 and persisted up to adulthood. Double labelling experiments using a monoclonal antibody against the proliferating cell nuclear antigen (PCNA), a cyclin synthesised by mitotic cells, showed that neuronal and/or glial polypeptides are expressed only by fully differentiated postmitotic cells. These results indicate that major events in the neurochemical maturation of the rabbit cerebellum occur during the first month after birth, when the same pattern of the adult animal is attained.

Mechanism of action of the sodium pump in vertebrate photoreceptors.

Membrane potential and photoresponse of rods in the isolated toad retina was recorded while changing the ionic composition of the extracellular medium. In the presence of 10 mM external Cs+ the bright flash response consisted of an initial fast component of about 35 mV followed by a much slower component as large as 50 mV. The slow component of the photoresponse was blocked by micromolar amounts of both ouabain and strophanthidin. The effects induced by the latter were almost completely irreversible. The effects induced by changing the external concentrations of Na+, K+ and Ca2+ were analysed assuming that the amplitude of the slow component of voltage photoresponse reflects the activity of Na+--K+ pump. It is shown that external Na+, K+ and Ca2+ have the same effect on the amplitude of the slow component as on the Na+-K+ pump activity of other tissues.

Histochemical research into the facial caruncle of the Muscovy duck (Cairina moschata domestica L.) and its relationship with the adrenal glands and gonads.

Histochemical studies carried out on prepuberal and adult, males and females, Muscovy ducks (the Black ancestral variety) showed that the beginning of the facial caruncle is caused mainly by local androgenic activities. In adult males, support for this organ comes from the steroid hormones originating from the adrenal gland and testis. In adult females, caruncular androgenic activity also seems to be necessary for the support of this organ.

Diurnal changes in the pigeon electroretinogram.

The spectral sensitivity of the pigeon Electroretinogram was determined at different times during the 24-hour cycle. These measures show that diurnal changes occur in the relative sensitivity of the electroretino-graphic responses to light stimuli of different wavelength. Diurnal variations occur despite the absence of rhythmic photoperiodic stimulation. It is suggested that a spontaneous shift occurs in the functional dominance of cones and rods with a period near to 24 hours.