Microwave ablation for large benign thyroid nodules: a proposal for a new approach: "the fluid-motion technique".

INTRODUCTION: Most thyroid nodules are benign, and treatment is primarily indicated for patients with symptoms or cosmetic concerns related to nodular volume. Recently, there has been growing interest in nonsurgical and minimally invasive techniques for the treatment of symptomatic benign thyroid nodules, including microwave ablation (MWA). AIMS: This study aims to evaluate the effectiveness, efficacy, and safety of ultrasound-guided uncooled MWA in the treatment of benign symptomatic thyroid nodules, using a new ablation technique called the "fluid-motion technique." MATERIALS AND METHODS: From April 2021 to February 2023, a total of 150 patients with benign thyroid nodules were treated using MWA at the Endocrine Surgery Unit of Pederzoli Hospital, Peschiera del Garda (Italy), with 102 of them being followed-up for at least 6 months. RESULTS: Before treatment, the mean major diameter was 38.4 ± 10.0 mm, and the mean volume was 13.6 ± 10.5 mL. At 1 month, the mean volume was 6.8 ± 6.2 mL, and at 3 months, it was 4.6 ± 4.1 mL. The mean Volume Reduction Rate (VRR) at 3 months was 61.1 ± 22.0%. Of the nodules, 17 (17%), 65 (66%), and 20 (19%) had volumes ≤ 10, 10.1-20, and ≥ 20.1 mL, respectively. For these nodules, the mean VRR at 3 and 6 months was 55.3 ± 16%, 63.4 ± 24.8%, 72 ± 26.9% and 68.0 ± 11.2%, 73.5 ± 18.3%, and 81.6 ± 15.8%, respectively. CONCLUSION: Our study shows that ultrasound-guided MWA is an effective and safe minimally invasive treatment for benign thyroid nodules. The goal is to achieve a VRR that can reduce symptoms and improve cosmetic appearance.

Gestación en paciente con insuficiencia ovárica primaria secundaria a mosaicismo de Síndrome de Turner / Pregnancy in a patient with primary ovarian insufficiency owing to a mosaicism of Turner Syndrome

RESUMEN La Insuficiencia Ovárica Primaria se define por una amenorrea secundaria de al menos cuatro meses de duración, deficiencia de esteroides sexuales (estradiol) y altas concentraciones séricas de hormona folículoestimulante (FSH) con al menos un mes de diferencia entre estas determinaciones, en mujeres menores de 40 años. Es una causa insidiosa de infertilidad pero en algunas ocasiones es transitoria y permite una gestación espontánea. El Síndrome de Turner es un trastorno genético caracterizado por la pérdida o anomalías estructurales de un cromosoma X y que afecta a 1 de cada 2.500 mujeres nacidas vivas. Las manifestaciones clínicas varían entre pacientes, pero generalmente se relaciona con talla baja, coartación aórtica, disgenesia gonadal e insuficiencia ovárica primaria. Las técnicas de reproducción asistida como la criopreservación de ovocitos y de tejido ovárico, la maduración in vitro o la donación de ovocitos ofrecen opciones reproductivas en aquellos casos en los que no se produzca un embarazo espontáneo.

ABSTRACT Primary Ovarian Insufficiency is considered a secondary amenorrhea of at least four months duration, sex steroid deficiency (estradiol) and high serum concentrations of follicle stimulating hormone (FSH) with at least one month difference between these determinations, in women under 40 years. It is an insidious cause of infertility but sometimes it is transient and allows a spontaneous pregnancy. Turner syndrome is a genetic disorder characterized by the loss or structural abnormalities of an X chromosome that affects 1 in 2,500 women born alive. Clinical manifestations vary among patients, but it is usually associated with short stature, aortic coarctation, gonadal dysgenesis, and primary ovarian failure. Assisted reproduction techniques such as cryopreservation of oocytes and ovarian tissue, in vitro maturation or oocyte donation offer reproductive options in those cases in which there is no spontaneous pregnancy.

Informing the Adaptation and Implementation of a Lifestyle Modification Program in Hispanics: A Qualitative Study Among Low-Income Hispanic Adults.

Low-income Hispanics are a hard to reach population that face unique challenges in achieving behavior change. Understanding such challenges can inform the adaptation of lifestyle behavioral interventions and increase participant engagement. This descriptive qualitative study aimed to (1) inform lifestyle modification programs focused on improving the physical activity and healthy eating of low-income Hispanic adults and (2) explore potential gender differences necessary for consideration. Five gender-specific focus groups were conducted with Hispanic adults from a free community clinic (n = 45, 64% women, 53.1 ± 10.1 years). Sessions were audio-recorded, transcribed, and coded using a general inductive approach. The qualitative software program Dedoose was used to perform a content analysis. Emergent themes included topics of interest, methods of increasing engagement in healthy behaviors, and the delivery of lifestyle modification programs. Men expressed greater interest in physical activity, while women desired to learn about healthy eating. Men described the importance of physician communication, while women focused on the role of social support as facilitators for participating in such programs. Both genders described education as key to behavior change and viewed text messaging as a valuable complement. Results from this study can inform the adaptation of lifestyle modification programs and increase the engagement of low-income Hispanic populations.

Bone histomorphometry in acromegaly patients with fragility vertebral fractures.

CONTEXT: The high risk of vertebral fractures (VFs) in acromegaly patients despite normal bone mineral density (BMD) is well known. The reasons for this paradoxical finding of skeleton fragility are poorly understood due to the limited data on bone histomorphometry in acromegaly. OBJECTIVE: This study aimed to analyze histomorphometric parameters including bone microarchitecture in acromegaly patients with VFs and normal BMD compared to normal subjects, and also to evaluate the differences between active and controlled acromegaly patients. PATIENTS AND METHODS: Forty-seven acromegaly patients (17 active, 30 controlled), median (range) age 57 years (30-88) were evaluated for bone turnover, morphometric VFs and BMD by dual-energy X-ray absorptiometry at lumbar spine and hip; 12 patients with VFs and normal BMD underwent iliac crest bone biopsy; 12 biopsies were taken at the autopsy in healthy sex and age-matched control subjects. RESULTS: The histomorphometric evaluation of acromegaly fractured patients was compared with that of normal controls and showed significantly reduced median (range) levels of bone volume/tissue volume (BV/TV: 15.37% (7.93-26.75) vs. 18.61% (11.75-27.31), p = 0.036), trabecular thickness (TbTh: 77.6 µm (61.7-88.3) vs. 82.7 µm (72.3-92.0) p = 0.045), with increased trabecular separation (TbSp: 536.4 µm (356.2-900.6) vs. 370.3 µm (377.1-546.3) p = 0.038) and increased cortical thickness (1268 µm (752-2521) vs. 1065 µm (851-1205) p = 0.025) and porosity (11.9% (10.2-13.3) vs. 4.8% (1.6-8.8) p = 0.0008). While active acromegaly patients showed histomophometric features of increased bone turnover, patients with controlled disease presented normal bone turnover with significantly lower osteoblastic activity, expressed as osteoblast number (p = 0.001), active osteoblasts and vigor (p = 0.014) in the presence of reduced osteocyte number (p = 0.008) compared to active disease. CONCLUSIONS: The apparent paradox of bone fragility in acromegaly patients with a normal BMD can be explained by increased cortical thickness and porosity and reduced trabecular thickness with increased trabecular separation. These structural and microarchitectural abnormalities persist in the controlled phase of acromegaly despite bone turnover normalization. The main determinant of bone disease after hormonal control is severe osteoblastic dysfunction.

Lymphnode metastasis of thyroid cancer misinterpreted as lateral aberrant thyroid 40 years before identification of primary tumor. Case report and review of the literature.

The differential diagnosis between lateral ectopic thyroid tissue with orthotopic normal gland and metastatic thyroid carcinoma is challenging. Lateral cervical site is a very rare location for ectopic tissue since only a few cases have been reported. The peculiarity of this clinical case is the finding of a thyroid carcinoma forty years after surgical resection of the ectopic thyroid lesion. This asynchronous association, never reported in literature, raises the question of the differential diagnosis between a true ectopic aberrant thyroid and an early lymph node metastasis from an occult thyroid carcinoma, evident in the primitive site many years later. Several elements, which will be matter of discussion, seem to favour the latter hypothesis.This case, although isolated, suggests that any lateral cervical mass, comprising thyroid tissue, should be regarded as a metastasis of thyroid carcinoma until proven otherwise. Carefull investigation of thyroid gland is mandatory.

Study of the correlation between sensing performance and surface morphology of inkjet-printed aqueous graphene-based chemiresistors for NO2 detection.

The extremely high sensitivity to the external environment and the high specific surface area, as well as the absence of bulk phenomena that could interfere with the response signal, make graphene highly attractive for the applications in the field of sensing. Among the various methods for producing graphene over large areas, liquid phase exfoliation (LPE) appears to be very promising, especially if combined with inkjet printing (IJP), which offers several advantages, including the selective and controlled deposition of small ink volumes and the versatility of the exploitable inks and substrates. Herein we present a feasibility study of chemiresistive gas sensors inkjet-printed onto paper substrates, in which a LPE graphene suspension dispersed in a water/isopropanol (H2O/IPA) mixture is used as sensing ink. The device performances, in terms of relative conductance variations, upon exposure to NO2 at standard ambient temperature and pressure, are analysed. In addition, we examine the effect of the substrate morphology and, more specifically, of the ink/substrate interaction on the device performances, by comparing the response of different chemiresistors fabricated by dispensing the same suspension also onto Al2O3 and Si/SiO2 substrates and carrying out a supportive atomic force microscopy analysis. The results prove the possibility to produce sensor devices by means of a wholly environmentally friendly, low-cost process that meets the requests coming from the increasing field of paper-based electronics and paving the way towards a flexible, green-by-design mass production.

Androgen actions in mouse wound healing: Minimal in vivo effects of local antiandrogen delivery.

The aims of this work were to define the role of androgens in female wound healing and to develop and characterize a novel wound dressing with antiandrogens. Androgens retard wound healing in males, but their role in female wound healing has not been established. To understand androgen receptor (AR)-mediated androgen actions in male and female wound healing, we utilized the global AR knockout (ARKO) mouse model, with a mutated AR deleting the second zinc finger to disrupt DNA binding and transcriptional activation. AR inactivation enhanced wound healing rate in males by increasing re-epithelialization and collagen deposition even when wound contraction was eliminated. Cell proliferation and migration in ARKO male fibroblasts was significantly increased compared with wild-type (WT) fibroblasts. However, ARKO females showed a similar healing rate compared to WT females. To exploit local antiandrogen effects in wound healing, while minimizing off-target systemic effects, we developed a novel electrospun polycaprolactone (PCL) scaffold wound dressing material for sustained local antiandrogen delivery. Using the antiandrogen hydroxyl flutamide (HF) at 1, 5, and 10 mg/mL in PCL scaffolds, controlled HF delivery over 21 days significantly enhanced in vitro cell proliferation of human dermal fibroblasts and human keratinocytes. HF-PCL scaffolds also promoted in vivo wound healing in mice compared with open wounds but not to PCL scaffolds.

The diverse toxic effect of SiO2 and TiO2 nanoparticles toward the marine microalgae Dunaliella tertiolecta.

Nanoparticles (NPs) are widely used in many industrial applications. NP fate and behavior in seawater are a very important issue for the assessment of their environmental impact and potential toxicity. In this study, the toxic effects of two nanomaterials, silicon dioxide (SiO2) and titanium dioxide (TiO2) NPs with similar primary size (~20 nm), on marine microalgae Dunaliella tertiolecta were investigated and compared. The dispersion behavior of SiO2 and TiO2 NPs in seawater matrix was investigated together with the relative trend of the exposed algal population growth. SiO2 aggregates rapidly reached a constant size (600 nm) irrespective of the concentration while TiO2 NP aggregates grew up to 4 ± 5 µm. The dose-response curve and population growth rate alteration of marine alga D. tertiolecta were evaluated showing that the algal population was clearly affected by the presence of TiO2 NPs. These particles showed effects on 50 % of the population at 24.10 [19.38-25.43] mg L(-1) (EC50) and a no observed effect concentration (NOEC) at 7.5 mg L(-1). The 1 % effect concentration (EC1) value was nearly above the actual estimated environmental concentration in the aquatic environment. SiO2 NPs were less toxic than TiO2 for D. tertiolecta, with EC50 and NOEC values one order of magnitude higher. The overall toxic action seemed due to the contact between aggregates and cell surfaces, but while for SiO2 a direct action upon membrane integrity could be observed after the third day of exposure, TiO2 seemed to exert its toxic action in the first hours of exposure, mostly via cell entrapment and agglomeration.

VEGF-A polymorphisms predict progression-free survival among advanced castration-resistant prostate cancer patients treated with metronomic cyclophosphamide.

BACKGROUND: No data are available on the pharmacogenetics of metronomic chemotherapy in prostate cancer. The aim of this study was to evaluate the association between VEGF-A sequence variants and prostate-specific antigen (PSA) progression, progression-free survival (PFS) and overall survival (OS), in advanced castration-resistant prostate cancer patients treated with metronomic cyclophosphamide (CTX), celecoxib and dexamethasone. METHODS: Forty-three patients were enrolled, and genomic DNA was extracted. VEGF-A gene SNPs (-2578A/C, -634C/G, +936C/T) were analysed using TaqMan PCR assays. Hardy-Weinberg equilibrium was tested for each SNP, and genetic effects were evaluated by Fisher's exact test. PFS and OS were analysed with GraphPad Prism software, using the product limit method of Kaplan and Meier, and comparing survival curves using both the log-rank test and the Gehan-Wilcoxon test. We used Bonferroni correction to account for multiple testing, and a two-tailed P-value of <0.017 was considered statistically significant. RESULTS: Overall, 20 patients (46%) experienced a reduction in PSA levels from baseline and, among them, 14 (32%) showed a confirmed PSA ≥50% decrease. In non-responders, the -2578CC genotype was more frequent (18.60% vs 2.33% in responders; P=0.0212) whereas the -634CC genotype frequency was 22.73% vs 0% in responders (P=0.0485). With regard to PFS, patients harbouring the -634CC genotype had a median PFS of 2.2 months whereas patients with the genotype -634CG/GG had a median PFS of 6.25 months (P=0.0042). CONCLUSION: The -634CC genotype is significantly associated with a shorter PFS in patients treated with a metronomic CTX schedule.

Presentation, diagnostic features and glucose handling in a monocentric series of insulinomas.

BACKGROUND: New aspects have emerged in the clinical and diagnostic scenarios of insulinoma: current guidelines have lowered the diagnostic insulin threshold to 3 µU/ml in the presence of hypoglycemia (<55 mg/dl); post-prandial hypoglycemia has been reported as the only presenting symptom; preexisting diabetes mellitus (DM) was recognized in some patients. AIM: To evaluate clinical features, diagnostic criteria and glucose metabolic profile in a monocentric series of patients affected by insulinomas including two subgroups: sporadic and multiple endocrine neoplasia type-1 syndrome (MEN-1). SUBJECTS AND METHODS: Clinical, pathological and biochemical data regarding 33 patients were analyzed. RESULTS: following the current guidelines the 72-h fasting test was initially positive in all cases but one. In this case the test, initially negative, became positive after a 2-yr follow-up. Nadir insulin level was ≥ 3 µU/ml but <6 µU/ml in 3 patients and ≥ 6 µU/ml in the remaining 30 cases. At presentation, 27 patients (82%) reported only fasting symptoms, 3 (9%) only post-prandial and 3 (9%) both. Seven cases (21%) had previously been affected by type 2 DM or impaired glucose metabolism. CONCLUSIONS: In our series the new cut-off of insulin increased the sensitivity of the 72-h fasting test from 87% to 97%. The absence of hypoglycemia during the test cannot definitively rule out the diagnosis and the test should be repeated in every highly suspicious case. Post-prandial hypoglycemia can be the only presenting symptom. DM may be associated with the occurrence of insulinoma. So that a possible diagnosis of insulinoma must not be ignored if previous impaired glucose handling is evident.

Natural history of gastro-entero-pancreatic and thoracic neuroendocrine tumors. Data from a large prospective and retrospective Italian epidemiological study: the NET management study.

BACKGROUND: The few epidemiological data available in literature on neuroendocrine tumors (NET) are mainly based on Registry databases, missing therefore details on their clinical and natural history. AIM: To investigate epidemiology, clinical presentation, and natural history of NET. DESIGN AND SETTING: A large national retrospective survey was conducted in 13 Italian referral centers. Among 1203 NET, 820 originating in the thorax (T-NET), in the gastro-enteropancreatic tract (GEP-NET) or metastatic NET of unknown primary origin (U-NET) were enrolled in the study. RESULTS: 93% had a sporadic and 7% a multiple endocrine neoplasia type 1 (MEN1)-associated tumor; 63% were GEP-NET, 33% T-NET, 4% U-NET. Pancreas and lung were the commonest primary sites. Poorly differentiated carcinomas were <10%, all sporadic. The incidence of NET had a linear increase from 1990 to 2007 in all the centers. The mean age at diagnosis was 60.0 ± 16.4 yr, significantly anticipated in MEN1 patients (47.7 ± 16.5 yr). Association with cigarette smoking and other non-NET cancer were more prevalent than in the general Italian population. The first symptoms of the disease were related to tumor burden in 46%, endocrine syndrome in 23%, while the diagnosis was fortuity in 29%. Insulin (37%) and serotonin (35%) were the most common hormonal hypersecretions. An advanced tumor stage was found in 42%, more frequently in the gut and thymus. No differences in the overall survival was observed between T-NET and GEP-NET and between sporadic and MEN1-associated tumors at 10 yr from diagnosis, while survival probability was dramatically reduced in U-NET. CONCLUSIONS: The data obtained from this study furnish relevant information on epidemiology, natural history, and clinico-pathological features of NET, not available from the few published Register studies.

Carcinoid crisis induced by receptor radionuclide therapy with 90Y-DOTATOC in a case of liver metastases from bronchial neuroendocrine tumor (atypical carcinoid).

SS receptors are overexpressed in many tumors, mainly of neuroendocrine origin, thus enabling the treatment with SS analogs. The clinical experience of receptor radionuclide therapy with the new analog [90Y-DOTA0-Tyr3 ]-octreotide [90Y-DOTATOC] has been developed over the last decade and is gaining a pivotal role in the therapeutic workout of these tumors. It is well known that some procedures performed in diagnostic and therapeutic management of endocrine tumors, such as agobiopsy and hepatic chemoembolization, can be associated with the occurrence of symptoms related to the release of vasoactive amines and/or hormonal peptides from tumor cell lysis. This is the first report of a severe carcinoid crisis developed after receptor radionuclide therapy with 90Y-DOTATOC administered in a patient affected by liver metastases from bronchial neuroendocrine tumor (atypical carcinoid). Despite protection with H1 receptor antagonists, octreotide and corticosteroids, few days after the therapy the patient complained of persistent flushing of the face and upper trunk, severe labial and periocular oedema, diarrhoea and loss of appetite. These symptoms increased and required new hospitalisation. The patient received iv infusion of octreotide associated with H1 and H2 receptor antagonists and corticosteroid therapy, which induced symptom remission within few days. The case here reported confirms that radionuclide therapy is highly effective in determining early rupture of metastatic tissue and also suggests that pre-medication should be implemented before the radiopeptide administration associated with a close monitoring of the patient in the following days.

Long-term quiescence of ectopic Cushing's syndrome caused by pulmonary neuroendocrine tumor (typical carcinoid) and tumorlets: spontaneous remission or therapeutic effect of bromocriptine?

In 1990, a 55-yr-old woman was admitted to the Medical Department of our hospital for severe hypercortisolism complicated by secondary diabetes mellitus and serious hypokalemia. Although inferior petrosal sinus sampling did not show any significant difference between central and peripheral ACTH concentration, suggesting an ectopic source of ACTH secretion, diagnostic imaging was negative and Cushing's disease due to hyperplasia of the pituitary intermediate lobe was suspected. Medical treatment with bromocriptine and cyproheptadine led to a rapid and stabile normalization of adrenal function, so that after two months cyproheptadine was stopped and bromocriptine was tapered to a smaller dose. An attempt to discontinue medical treatment, carried out 3 yr later, was followed by a quick increase of ACTH and cortisol levels, which were normalized by the resumption of the bromocriptine. Adrenal function remained normal until 1994 when hypercortisolism relapsed despite the treatment. Chest radiography and computed tomography (CT) scan detected a 6 mm nodule in the middle lobe of the lung which proved to be a neuroendocrine tumor, with immunohistochemical positivity for ACTH. Nests of neuroendocrine cells (tumorlets) were also demonstrated in the surrounding lung tissue. After the lobectomy, the patient recovered completely from Cushing's syndrome and no symptoms and/or signs of recurrence have been observed over the subsequent follow-up period. Although cyclical spontaneous Cushing's syndrome could not be excluded, there was strong evidence that medical treatment with bromocriptine might have played a key role in long-lasting remission. To our knowledge, this is the second case described in literature of Cushing's syndrome caused by neuroendocrine lung tumor responsive to bromocriptine.

Thyroxine hair content in congenital hypothyroidism and hyperthyroidism.

Using the determination of thyroxine (T4) hair content, we studied 16 hypothyroid newborns diagnosed by means of our regional screening program, and five hypothyroid infants, undetected at birth, at diagnosis and after 3 months of substitutive therapy (8-10 microg/kg/day L-thyroxine in newborns; 15 microg/kg/day in infants), and 13 hyperthyroid adults. Hair T4 content was similar at diagnosis in hypothyroid newborns (2.6 +/- 2.3 pg/mg hair) and in infants undetected at birth (2.4 +/- 1.7 microg/mg hair), but very high only in the latter after therapy (23.2 +/- 3.9 microg/mg hair). Untreated hyperthyroid adults surprisingly evidenced lower hair T4 (0.4 +/- 0.2 microg/mg hair) than controls (1.5 +/- 0.3 microg/mg hair). We suggest these findings are due to differential tissue storage of thyroid hormone, related to the different blood T4 concentration. Therefore, T4 hair assay could be a non-invasive method to further assess thyroid status.

Differential display.

The study of cancer genetics has focused primarily, until now, on changes at the genomic level. However, activating, or inactivating, mutations/deletions/rearran ments in DNA are not the only way in which alterations in gene expression can occur. Since the development of subtractive hybridization, which screens for differentially expressed genes between related populations at the RNA level, evidence has accrued showing that alterations in downstream gene expression might play a significant role in the biology and evolution of cancer. Establishment of the novel technique of differential display (DD) 1,2(1,2) has facilitated studies in this new field that has been termed "expression genetics" 3). Compared with traditional methods of gene expression analysis DD possesses the advantages of simplicity, sensitivity, reproducibility, and speed. It is the current method of choice in expression genetics not only in the cancer biology field but also in such diverse areas as embryology, developmental biology, and neurobiology. This novel technique involves the reverse transcription (RT) of mRNAs with oligo-dT primers anchored to the beginning of the poly(A) tail, followed by a poly merase chain reaction (PCR) in the presence of a second arbitrary random primer. The amplified cDNAs of 3' termini of mRNAs obtained (with the same primers) from different samples can then be run side by side on a sequencing gel to allow differentially expressed genes, either upregulated or downregulated, to be visualized (see Fig. 1). By changing primer pair combinations approx 15,000 individual mRNA species from a mammalian cell may be analyzed reasonably quickly, thus providing a "fingerprint" of the mRNA in any particular cell type. Differential cDNA fragments can easily be recovered and reamplified from the gel and cloned into vectors for future screening and characterization work. The DD technique can be subdivided into five steps fig. 1. Principal procedures of differential display technique.

Identification of changes in gene expression associated with minimal residual disease.

Minimal residual disease (MRD), the tumour burden which remains after a course of treatment that has resulted in clinical remission [1], appears to differ in certain characteristics from the primary tumour population. Certainly the cells which comprise MRD have had to escape from the constraints of the primary tumour mass, invade normal tissue and penetrate small vessels in order to enter the circulation in which they then have had to survive. Such activities are the consequence of the expression of specific proteins and these may well be a reflection of alterations in DNA or RNA levels. Identifying the changes in RNA expression levels between related cell groups exhibiting different phenotypes recently has become a great deal easier as a consequence of developments in analytical procedures such as Differential Display (DD) and Serial Analysis of Gene Expression (SAGE). Application of these procedures to MRD cells recovered from blood, bone marrow or lymph node, should identify novel sequences associated with tumour progression and the development of disseminated disease.

Identification by differential display of a protein phosphatase-2A regulatory subunit preferentially expressed in malignant melanoma cells.

We described the occurrence of 4 transcripts differentially displayed between syngeneic murine B16F10 (metastatic melanoma) and Melan-a (immortalised melanocytes) cell lines. We now report that one such transcript, which is B16F10-specific, represents a protein phosphatase-2A B' regulatory subunit. No expression of this transcript was detected in the weakly metastatic B16F1 by Northern blotting. Moreover, the transcript was not expressed by spontaneously immortalised, non-tumorigenic, melanocytes (Melan-Ab and Melan-a2), nor was it expressed by ras-transformed, tumourigenic melanocytes (Melan-Ab-LTR-ras). Cloning of the 5'-end region of this transcript (termed band 8A) from B16F10 cells revealed an intracisternal A-particle insertion, including the long terminal repeat region, which could account for the observed high expression in B16F10 cells. Single cell clones of B16F10 manifested an experimental metastasis capacity, which correlated with band 8A expression with the lowest expressors being least metastatic. The human homologue of the B' regulatory subunit, B56gamma, is expressed preferentially at the mRNA level in human melanoma cell lines compared with normal epidermal melanocytes. In situ hybridisation studies on human clinical samples detected high expression of this gene in a number of malignant melanomas. Our results imply strongly that this protein phosphatase-2A regulatory subunit may have a role in melanoma tumour progression.

The TSH-dependent variation of the essential elements iodine, selenium and zinc within human thyroid tissues.

Instrumental Neutron Activation Analysis was used in order to measure iodine, selenium and zinc concentration in thyroid samples. A pair of samples of normal and nodular tissue were collected from the thyroid gland from 72 patients selected on the basis of pathological criteria (44 cases of multinodular goiter, 12 of chronic lymphocytic thyroiditis (CLT), 6 of thyroid adenoma (TA) and 12 of thyroid cancer (TC)). The check for tissue homogeneity and sampling error was performed by means of the coefficient of variation (CV%) of the elements in replicate samples of normal and altered tissues. High CV% values (> 15%) for iodine reflected a functional variability in thyroid follicles, while low CV% values (< 10%) for selenium and zinc indicated that the composition of selected tissues was rather homogeneous. The variation of the element's concentration was compared in normal and altered tissues. The mean element concentrations had values close to those already reported in the literature; furthermore, our patients had marginal iodine and selenium deficiency. Both normal and nodular tissues in CLT showed statistically significant lower zinc values as compared with the other thyroid diseases. To evaluate the thyroid function, thyroid stimulating hormone (TSH) and thyroxine (T4) levels were measured in the serum of patients. Two arbitrary serum-TSH threshold levels (TSH < 1.0 and > 4.0 mU/L) were introduced in order to classify, respectively, hyperthyroidism and hypothyroidism, as well as euthyroid conditions (1.0 < TSH < 4.0 mU/L), and each patient was assigned to one of these groups. The influence of TSH in the variation of the concentration of iodine, selenium and zinc in normal and altered human thyroid tissues was significant.

Heterogeneity of nuclear DNA pattern and its relationship with cell cycle activity parameters in multinodular goitre.

OBJECTIVE: Recent studies suggest that the malignancy rate in multinodular goitre is not significantly different from that observed in solitary nodules and that chromosomal aberrations are not infrequent in multinodular goitre. To further investigate this topic we determined the DNA pattern in multinodular goitres. DESIGN: DNA ploidy and cell cycle activity parameters were determined in multinodular goitres. PATIENTS: We evaluated 235 patients (185 female, 50 male, mean age 52 +/- 13 years), who had undergone thyroidectomy; 11 of them harboured occult differentiated microcarcinoma. MEASUREMENTS: DNA index (DI), coefficient of variation of G0/G1 phase (CV), percentage of cells in S phase (%S) and in G2+M phase (%G2-M) and proliferative index (PI = %S+%G2-M) were determined by flow cytometric analysis (FCM) in tissue samples taken from 3 different areas of the thyroid gland. RESULTS: Aneuploid DNA was found in 50 goitres without carcinoma (22.3%) and in 5 goitres with carcinoma (45.5%). The mean PI of euploid cells in the goitre without carcinoma was significantly higher in the goitres with an aneuploid component compared to the goitres without aneuploidy (10.8 +/- 1.3 SEM vs 6 +/- 0.32; P +/- 0.001). Also, the percentage difference between maximal and minimal PI found within each goitre (delta PI %) was higher in the former group (373 +/- 49 SEM vs 142 +/- 11.3; P < 0.0001). The PI was significantly higher in goitres with carcinoma compared to the goitres without carcinoma (12.9 +/- 3.2 SEM vs 7.07 +/- 0.40; P < 0.05). CONCLUSIONS: The findings of increased proliferation rate in goitres with an aneuploid or neoplastic component suggests that some factors involved in goitrogenesis could also be responsible for the development of chromosomal aberrations and/or for the selection of cellular clones endowed with high growth potential.

[Cushing's disease associated with empty sella: a clinical case treated for years with ketoconazole]. / Malattia di Cushing associata a sella vuota: un caso clinico trattato per sette anni con ketoconazolo.

Cushing's disease and empty sella without evidence of pituitary adenoma are rarely observed. To our knowledge, there is very little documentation on long-term therapeutic follow-up with the steroidogenesis inhibitor ketoconazole. A 48-year-old woman with uncontrolled insulin-dependent diabetes mellitus, severe hypertension, and clinical findings of hypercortisolism was referred to our hospital. Endocrine evaluation of adrenocortical function evidenced hypothalamic-pituitary-hypercortisolism, and excluded adrenal tumor or an ectopic corticotropin source. Magnetic resonance imaging disclosed an empty sella turcica but not pituitary adenoma. The patient was treated with a steroidogenesis inhibitor, ketoconazole (600 mg daily) which reduced urinary cortisol excretion to within the normal range. Serum cortisol levels also returned to normal in the morning but not in the evening. The patient has continued on ketoconazole therapy for the past 7 years, with neither side effects nor tachyphylaxis. The reduction of cortisol secretion brought about significantly improved control of diabetes mellitus and hypertension, although signs of hypercortisolism have persisted. Radiographic studies of the hypophysis during follow-up have not evidenced adenoma.