Angela Mariano

A mouse model for hereditary thyroid dysgenesis and cleft palate.

Alteration of thyroid gland morphogenesis (thyroid dysgenesis) is a frequent human malformation. Among the one in three to four thousand newborns in which congenital hypothyroidism is detected, 80% have either an ectopic, small and sublingual thyroid, or have no thyroid tissue. Most of these cases appear sporadically, although a few cases of recurring familial thyroid dysgenesis have been described. The lack of evidence for hereditary thyroid dysgenesis may be due to the severity of the hypothyroid phenotype. Neonatal screening and early thyroid hormone therapy have eliminated most of the clinical consequences of hypothyroidism such that the heritability of this condition may become apparent in the near future. We have recently cloned cDNA encoding a forkhead domain-containing transcription factor, TTF-2, and have located the position of the gene, designated Titf2, to mouse chromosome 4 (ref. 3). Titf2 is expressed in the developing thyroid, in most of the foregut endoderm and in craniopharyngeal ectoderm, including Rathkes pouch. Expression of Titf2 in thyroid cell precursors is down-regulated as they cease migration, suggesting that this factor is involved in the process of thyroid gland morphogenesis. Here we show that Titf2-null mutant mice exhibit cleft palate and either a sublingual or completely absent thyroid gland. Thus, mutation of Titf2 –/– results in neonatal hypothyroidism that shows similarity to thyroid dysgenesis in humans.

MATERNAL SERUM AND UMBILICAL CORD BLOOD LEPTIN CONCENTRATIONS WITH FETAL GROWTH RESTRICTION

OBJECTIVE To ascertain whether fetal growth restriction is associated with alterations of leptin concentrations in umbilical cord blood and maternal serum. METHODS Maternal serum and umbilical cord blood leptin concentrations were determined by immunoradiometric assay at term in 43 women with uncomplicated singleton pregnancies (group A) and in 27 women with singleton pregnancies complicated by fetal growth restriction (group B), all with normal pregravid body mass index (BMI). RESULTS Maternal serum leptin concentrations were significantly higher in group B compared with group A (45.0 ng/mL [range 34.2–54.9] versus 29.0 ng/mL [range 24.7–33.3]; P < .01). Umbilical cord blood leptin levels were significantly lower in group B compared with group A (8.4 ng/mL [range 3.6–13.2] versus 13.1 ng/mL [9.7–16.5]; P < .01). Maternal serum leptin levels were not significantly correlated with maternal BMI or with neonatal birth weight in either group. Umbilical cord blood leptin concentrations were significantly correlated with neonatal birth weight in both groups. CONCLUSION Growth restricted fetuses at term show umbilical cord blood leptin concentrations significantly lower than those in normal fetuses, suggesting that fetal adipose tissue is a major source of leptin. Maternal serum leptin concentrations are higher in the presence of a growth restricted fetus. This increase might be due to an intrinsic placental mechanism, by which small placentas produce more leptin as a compensatory mechanism, or to early hypoxia.

Predicting prostate biopsy outcome: prostate health index (phi) and prostate cancer antigen 3 (PCA3) are useful biomarkers.

Indication for prostate biopsy is presently mainly based on prostate-specific antigen (PSA) serum levels and digital-rectal examination (DRE). In view of the unsatisfactory accuracy of these two diagnostic exams, research has focused on novel markers to improve pre-biopsy prostate cancer detection, such as phi and PCA3. The purpose of this prospective study was to assess the diagnostic accuracy of phi and PCA3 for prostate cancer using biopsy as gold standard. Phi index (Beckman coulter immunoassay), PCA3 score (Progensa PCA3 assay) and other established biomarkers (tPSA, fPSA and %fPSA) were assessed before a 18-core prostate biopsy in a group of 251 subjects at their first biopsy. Values of %p2PSA and phi were significantly higher in patients with PCa compared with PCa-negative group (p<0.001) and also compared with high grade prostatic intraepithelial neoplasia (HGPIN) (p<0.001). PCA3 score values were significantly higher in PCa compared with PCa-negative subjects (p<0.001) and in HGPIN vs PCa-negative patients (p<0.001). ROC curve analysis showed that %p2PSA, phi and PCA3 are predictive of malignancy. In conclusion, %p2PSA, phi and PCA3 may predict a diagnosis of PCa in men undergoing their first prostate biopsy. PCA3 score is more useful in discriminating between HGPIN and non-cancer.

Epidermal growth factor receptor and thyrotropin response in human thyroid tissues

The content of epidermal-growth-factor receptor (EGFr) and its relation to TSH-response were examined in 27 malignant thyroid tumors (5 follicular, 6 papillary, 5 medullary, 11 anaplastic carcinomas) and in 30 tumor-like lesions (21 hyperplastic goiters and 9 toxic adenomatous goiters). Normal 12 thyroid tissues adjacent to benign tumors with no evidence of macroscopic or microscopic abnormalities were used as control. All thyroid plasma membranes tested showed specific EGF binding. In membranes from toxic adenomas (2.18 ± 0.73 fmoles/mg protein) and papillary carcinomas (2.80 ± 0.80) the EGF binding was similar to that of normal thyroid membranes (2.32 ± 0.73). Hyperplastic goiters showed an EGF binding (4.4 ± 0.82) slightly higher than normal tissue. The highest and the lowest EGF binding values were found in anaplastic (11.8 ± 2.78) and medullary (0.50 ± 0.39) carcinomas, respectively. An inverse correlation between EGFr content and TSH-response was found when anaplastic thyroid tumors were compared to tumorlike lesions. However no correlation was observed in medullary carcinomas which also failed to respond to TSH and in papillary carcinomas which partially respond to thyrotropin.

CENTRAL PRECOCIOUS PUBERTY : A SINGLE BLOOD SAMPLE AFTER GONADOTROPIN-RELEASING HORMONE AGONIST ADMINISTRATION IN MONITORING TREATMENT

This study was designed to evaluate whether a single blood sample drawn after the home injection of a long-acting gonadotropin-releasing hormone (GnRH) agonist (GnRHa) in patients treated for central precocious puberty (CPP) could be a more simple and inexpensive test with respect to the conventional GnRH stimulating test in assessing adequate suppression of the pituitary-gonadal axis. The response to the first therapeutic injection of the GnRHa triptorelin was studied in 14 newly diagnosed untreated females with CPP. The results were compared with the response that the same patients had to the conventional GnRH stimulation test performed at the time of diagnosis. A significant increase in LH, FSH and E2 levels was observed 12 h after the triptorelin intramuscular injection; serum peak values of LH (70.3 ± 58.5 IU/l), FSH (44.2 ± 21.7 IU/l) and E2 (489.7 ± 263.9 pmol/l) were significantly greater than those obtained with the conventional GnRH test (LH 31.4 ± 21.7, p = 0.002; FSH 19.8 ± 10.7, p = 0.001; E2 83.3 ± 25, p < 0.001). In particular, the E2 response, 12 h after triptorelin injection, was clearly consistent with gonadal activation compared to the modest E2 increase in response to the GnRH test. Thereafter 22 girls who were already being treated with triptorelin for CPP were evaluated to see whether a single blood sample drawn 12 h after the therapeutic home injection of GnRHa could be informative in assessing adequate suppression of the pituitary-gonadal axis. This response was also compared to the conventional GnRH stimulation test performed 2 days before the therapeutic triptorelin injection. In 7 girls with evidence of pubertal progression, the E2 response following the GnRHa injection (136.3 ± 44.4 pmol/l) was significantly higher with respect to the response after the GnRH stimulation test (73.0 ± 0.0; p < 0.02) indicating an inadequate suppression of the pituitary-gonadal axis. The present data suggest that a single blood sample drawn 12 h after the therapeutic home administration of triptorelin provides a simple, comfortable and inexpensive means of monitoring pituitary as well as gonadal function in girls treated for CPP.

Epidermal growth factor receptor in human brain tumors

The expression of epidermal growth factor receptor (EGF-R) was examined in 27 primary human brain tumors (7 glioblastomas, 10 astrocytomas, 5 oligodendrogliomas, 1 schwannoma, 1 ganglioneuroma, 1 medulloblastoma, 1 ependimoma, 1 histiocitic lymphoma), in 6 brain metastases from lung carcinomas and in 20 meningiomas. Peritumoral tissues histologically normal excised surgically along with a large tumor were used as control. All plasma membranes from brain tissues tested showed specific EGF binding. The EGF receptor is expressed at low levels in the control human brain and at very high levels in 60% of the total intracranial tumors studied. When the various histological types of tumors were analyzed, the higher percentage of positive tumors was found with the meningiomas (85%) and the glioblastomas (71%), while the lower percentage of positivity was found with the oligodendrogliomas (40%) and the astrocytomas (30%). A good correlation between binding and total amount of EGF-R protein detected by Western Blot was also observed.

Endemic goiter in Calabria: Etiopathogenesis and thyroid function

An endemic goiter study was carried out in a vast territory of the Calabria region, including the provinces of Catanzaro and Cosenza. About 50% of 7231 school-children, aged between 6 and 12 years, examined in 34 villages, presented a thyroid enlargement. The high prevalence of goiter seems to be related to a moderate iodine deficiency intake indicated by a mild urinary iodine excretion (lower than 100 μg/g creatinine). In a few villages, in which a two year voluntary iodine prophylaxis was carried out, an increase in the urinary iodine excretion with a decreased goiter prevalence was observed. A slight increase in TT3, FT3 and TSH in one endemic area studied, compared to the control area, was also observed. These data suggest that an effective program of iodoprophylaxis is fundamental in this region as well.

Prognostic signs in the evolution of premature thelarche by discriminant analysis

Since premature thelarche (PT) can be a first sign of precocious puberty (PP), the aim of our study was to identify simple items in the course of the first 6 months of follow-up that could help predict if PT would evolve to PP. Thirty-two girls with PT were studied. First evaluation included bone age (BA), basal estradiol, FSH, LH and prolactin. GnRH was performed in 15 subjects and BA was checked at 6 month intervals in 30. Based on clinical outcome after a mean follow-up of 33.4 ± 16.5 (SD) months, patients were divided into 2 groups: Group I (G-I) included subjects whose breast development either remained unchanged, increased or regressed; Group II (G-II) included subjects who progressed to PP. The multivariate combination of the items which was able to best discriminate between the two groups was chosen in predicting the evolution of PT. The items considered included four variables available at the time of diagnosis [chronological (CA) at onset <3 years, basal FSH, basal LH and BA/CA ratio] and two additional variables after a 6-month follow-up (ΔBA/ΔCA and growth velocity); 88% of G-I and 14% of G-II had CA <3 yr. Basal FSH levels were elevated in both G-I (7.6 ± 3.0 mill/ml) and G-II (12.1 ± 4.1) with respect to controls (2.6 ± 1.2); however, approximately 20% of G-I had low FSH levels. Basal LH levels were consistently higher in G-II (8.0 ± 1.3 mlU/ml) than in G-I (2.9 ± 1.5) or controls (2.8 ± 1.2). Although initial BA was advanced (>2SD) in 21% of G-I and in all of G-II, an acceleration of BA was seen only in G-II. The mean growth velocity of G-I (44.1 ± 31.5%) was significantly less than G-II (92 ± 32%; p < 0.0025). With the help of the discriminant equations derived from data obtained at diagnosis and during the first 6 months of follow-up, all subjects with isolated premature thelarche could be sharply distinguished from those who subsequently progressed to precocious puberty. Such an equation should help in predicting the probability of considering either a diagnosis of isolated premature thelarche or precocious puberty in a child who presents with precocious breast development.

Gangliosides and phospholipids in human thyroids responsive and unresponsive to thyrotropin

The patterns of gangliosides and phospholipids and their relation to TSH response were examined in twenty-six malignant thyroid tumors (4 follicular, 6 papillary, 5 medullary, 11 anaplastic carcinomas) and thirty-six hyperplastic goiters. Thirteen thyroid tissues adjacent to benign tumors with no evidence of macroscopic or microscopic abnormalities were used as normal tissue. In normal thyroids the major ganglioside was GD3 (44%) and GM3 was the second ganglioside (20%). In minor amounts GD1a (8.6%), GD1b (6.2%), GT1b (5.7%) and GM1 (5.6%) were present. In hyperplastic goiters and in follicular carcinomas the patterns of gangliosides were similar to that of normal tissue except for GM3 which, in the last tissue, was higher (34%). In papillary carcinomas low levels of GM3 (11%) and GT1b (0.8%) with a high level of GD1b (12.6%) were found. In anaplastic carcinomas GM3 was very high (47%) whereas GD3 was low (18%). In these tumors also a high percentage (14.0%) of GD1a was found. In medullary carcinomas the lowest levels of GM3 (4%) with the highest level of GD3 (64%) were found. Although large differences of the gangliosides distribution were clearly encountered in the various pathological human thyroid, no correlation between lack of TSH response and some individual ganglioside could be made. No differences in the individual phospholipid in the various tissues studied were found.

Preoperative insulin-like growth factor-binding protein-3 (IGFBP-3) blood level predicts gleason sum upgrading.

About 43% of men with low Gleason grade prostate cancer (PCa) at biopsy will be finally diagnosed with high‐grade PCa at radical prostatectomy (RP). Gleason sum at RP is a good indicator of biochemical recurrence and poor clinical outcome. Therefore, there is a need to improve clinical evaluation of PCa aggressiveness in order to choice appropriate treatment. To this aim an easy‐available tool is represented by circulating biomarkers. Among these, the best candidates are some molecules involved in PCa pathogenesis such as IGFBP‐2 and IGFBP‐3, IL‐6, and its soluble receptor (SIL‐6R).