Localization of ανß6 integrin-TGF-ß1/Smad3, mTOR and PPARγ in experimental colorectal fibrosis.

A simultaneous action of several pro-fibrotic mediators appears relevant in the development of fibrosis. There are evidences that transforming growth factor-ß (TGF-ß)/Smad3 pathway forms with αvß6 integrin, mammalian target of Rapamycin (mTOR) and peroxisome proliferator-activated receptor-γ (PPARγ) a complex signalling network with extensive crosstalk and strong effects on fibrosis development. The present study evaluated the expression of TGFß, Smad3, αvß6 integrin, mTOR and PPARγ in 2,4,6-trinitrobenzenesulphonic acid (TNBS)-induced colorectal fibrosis in Smad3 wild-type (WT) and null mice. Smad3 WT mice treated with TNBS developed a marked colorectal fibrosis and showed a concomitant up-regulation of TGFß, Smad3, αvß6 and mTOR and a reduction of PPARγ expression. On the other hand, Smad3 Null mice similarly treated with TNBS did not develop fibrosis and showed a very low or even absent expression of TGFß, Smad3, αvß6 and mTOR and a marked over-expression of PPARγ. At the same time the expression of α-smooth muscle actin (a marker of activated myofibroblasts), collagen I-III and connective tissue growth factor (a downstream effector of TGFß/Smad3-induced extracellular matrix proteins) were up-regulated in Smad3 WT mice treated with TNBS compared to Null TNBS-treated mice. These preliminary results suggest a possible interaction between these pro-fibrotic molecules in the development of intestinal fibrosis.

Adenomyosis: from the sign to the diagnosis. Imaging, diagnostic pitfalls and differential diagnosis: a pictorial review.

Adenomyosis is a pathological gynaecological condition characterised by benign invasion of the endometrium into the myometrium. It is often misdiagnosed, or is not easily recognised, although it is responsible for disabling symptoms such as menorrhagia, abnormal uterine bleeding, dysmenorrhoea and infertility in premenopausal women. The aim of this pictorial review is to analyse the features of adenomyosis by illustrating the most usual and typical imaging patterns, along with the unusual appearances, seen in a vast array of gynaecological imaging modalities. The different findings of focal and diffuse adenomyosis along with the diagnostic limitations of ultrasound, hysterosalpingography and magnetic resonance imaging are described, as are the pitfalls and differential diagnosis with other pathological conditions that are often misdiagnosed as adenomyosis. The role of the different imaging modalities in planning appropriate treatment and their usefulness in monitoring therapy are also discussed.

Essential amino acids improve insulin activation of AKT/MTOR signaling in soleus muscle of aged rats.

Essential amino acids (EAA) improve basal muscle protein synthesis in the elderly. Nevertheless, in settings of prolonged supplementation, putative signal pathways of EAA are currently unknown. The purpose of this study was to test the effects of prolonged supplementation of EAA enriched mixture (12-L-Amin) on Insulin/Insulin-like Growth Factor-1 (IGF1) pathway by measuring total and phosphorylated Akt (Ser473) and its upstream (IRS1 at Ser636) and downstream (mTOR at Ser2448, p70S6K at Thr389) targets in basal conditions and following acute insulin (0.1 U/L) incubation in vitro. To this aim, soleus muscles were dissected from male Wistar rats divided in three groups of 7 each: adults (AD, 10 mo of age), elderly (EL, 22 mo of age) and elderly supplemented (EL-AA, 12-L-Amin 1.5gr/Kg die in drinking water for 3 mo). EL showed reduced basal and post-insulin mTOR and p70S6K activation and reduced post-insulin IRS1 degradation relative to AD. EL-AA showed an increase of post-insulin Akt activation, no change in basal and post-insulin phospho-mTOR, lower reduction of phospho-p70S6K and increased post-insulin IRS1 degradation relative to AD. These results demonstrate that chronic 12-LAmin administration exerts anti-ageing effects on the activation/inactivation of the Insulin/IGF1/mTOR pathway which is identified as putative target of EAA in the elderly.

Molecular aspects of gefitinib antiproliferative and pro-apoptotic effects in PTEN-positive and PTEN-negative prostate cancer cell lines.

To date, no effective therapeutic treatment allows abrogation of the progression of prostate cancer (PCa) to more invasive forms. One of the major targets for the therapy in PCa can be epidermal growth factor receptor (EGFR), which signals via the phosphoinositide 3'-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK) pathways, among others. Despite multiple reports of overexpression in PCa, the reliance on activated EGFR and its downstream signalling to the PI3K and/or MAPK/extracellular signal-regulated kinase (ERK) pathways has not been fully elucidated. We reported that the EGFR-selective tyrosine kinase inhibitor gefitinib (ZD1839; Iressa) is able to induce growth inhibition, G(1) arrest and apoptosis in PCa cells and that its effectiveness is associated primarily with phosphatase and tensin homologue deleted from chromosome 10 (PTEN) expression (and thus Akt activity). In fact PTEN-negative PCa cells are slowly sensitive to gefitinib treatment, because this molecule is unable to downregulate PI3K/Akt activity. PI3K inhibition, by LY294002 or after PTEN transfection, restores EGFR-stimulated Akt signalling and sensitizes the cells to pro-apoptotic action of gefitinib. The MAPK pathway seems to be involved primarily on cell-growth modulation because dual blockade of EGFR and ERK1/2 phosphorylation potentiates growth inhibition (both not cell apoptosis) in PTEN-positive PCa cells and reduced EGF-mediated growth in PTEN-negative cells. Thus the effectiveness of gefitinib requires growth factor receptor-stimulated PI3K/Akt and MAPK signalling to be intact and functional. The loss of the PTEN activity leads to uncoupling of this signalling pathway, determining a partial gefitinib resistance. Moreover, gefitinib sensitivity may be maintained in these cells through its inhibitory potential in MAPK/ERK pathway activity, modulating proliferative EGFR-triggered events. Therefore, our data suggest that the inhibition of EGFR signalling can result in a significant growth reduction and in increased apoptosis in EGFR-overexpressing PCa cells with different modalities, which are regulated by PTEN status, and this may have relevance in the clinical setting of PCa.

Uterine adenomyosis and tubal endometriosis: diagnostic imaging.

Endovaginal sonography plays a major role in the study of pelvic endometriosis and uterine adenomyosis. Other procedures as CT or MRI are reserved for particular cases. Diagnostic problems and main radiologic findings are analyzed.

Uterine intracavitary disorders: role of sonography and combination with other radiologic techniques.

Since it serves as the site for the implantation of the fertilized ovum, it is evident that significant disorders in the uterine cavity and endometrial mucosa represent potential factors of sterility/infertility. Among the acquired pathological conditions, a possible cause of sterility, there are inflammatory (endometritis) or post-traumatic (synechiae) disorders and proliferative mucosal (endometrial hyperplasia, polyps) and muscular (fibroma) disorders. Sonography (by transabdominal or preferably endovaginal route, and sonohysterography) is the first choice procedure being in most cases adequate for clinical assessment. According to the specific disease, the diagnostic combination with hysteroscopy, hysterosalpingography, Magnetic Resonance Imaging can be suitable.

Ovarian factor infertility.

The diseases of the ovary which most frequently cause infertility are: anovulation from follicular atresia, the empty follicle syndrome, the luteinized unruptured follicle syndrome; chronic anovulation syndromes, within which polycystic ovarian syndrome plays a major role; ovarian endometriosis. Sonography and Color Doppler US are the first choice procedures in the monitoring of ovarian cycles, which combined with serum hormone values, are able to identify possible changes in the physiologic sequence of the cycle. In follicular atresia, ovaries with minute follicles (3mm or less) and early disappearance of primary follicle are observed on sonography. The empty follicle syndrome characterized by the lack of oocytes within the primary follicle, is of difficult sonographic diagnosis, a possible sign being the missed visualization of cumulus oophorus. The luteinized unruptured follicle syndrome consists in the absence of oocyte expulsion from primary follicle persisting more than 48 hours after LH blood peak. Doppler spectra of blood flow in perifollicular ovarian arteries maintain the features of the follicular phase, i.e. low diastolic velocities and high resistances. Among chronic anovulation syndromes, hyper-and hypogonadotropism cause ovarian amenorrhea where ovaries are similar to those of women in menopause: small size, very few or absent follicles. The polycystic ovarian syndrome is characterized by an abnormal pulsatile GnRH release which causes LH hypersecretion and FSH hyposecretion. The latter is not able to stimulate the growth and maturation of follicles, while the former causes hyperandrogenism with hirsutism and obesity and is responsible for hypertrophy and stromal hyperechogenicity. The sonographic diagnosis of polycystic ovarian syndrome is based on standardized morphostructural signs as increased volume of the ovaries (> 10 cm3), the presence of numerous (> or = 10) peripheral microfollicles (< or = 5 mm) with hyperechoic stroma. The endometrial cyst, usually present in ovarian endometriosis is visualized with sonography as a round neoformation with ill-defined walls, filled with a uniformly hypoechoic, corpuscular, partly hemorrhagic fluid; less frequently the appearance is that of a more complex structure posing differential diagnostic problems, mainly with the hemorrhagic corpus luteum; both pathological conditions appear poorly vascularized at Color Doppler, with tracings of high resistance arterial flow. Among the procedures of second choice, CT can show the high blood density common to the two conditions while on MRI the signal is mostly hyperintense in T1-weighted sequences with areas of lower signal intensity in T2-weighted sequences.

Lipomatous tumors of the uterus: ultrasonographic findings in 11 cases.

We reviewed the ultrasonographic findings in eight patients with 11 lipomatous uterine tumors studied with transabdominal (eight cases) and transvaginal (six cases) techniques. A transvaginal color Doppler study was obtained in five patients. Two patients had more than one nodule (two and three uterine nodules, respectively). All lipomatous tumors had regular margins and were hyperechoic. Transvaginal sonography allowed the identification of one small previously undetected tumor but was not able to image the entire extent of two large lipomatous masses. In one case, it helped to establish the actual uterine origin of an eccentric pelvic mass. Color Doppler sonography showed complete absence of flow in all tumors examined. We believe that lipomatous tumors of the uterus can be diagnosed with a high degree of certainty by ultrasonography if a homogeneously hyperechoic avascular mass can be clearly identified to be of uterine origin.

[Experience with the combined diagnosis and therapy of locally advanced carcinoma of the uterine cervix (stage FIGO IIB-III). Transrectal ultrasonography and CT in the staging and in follow-up after therapy. Preliminary results]. / Esperienza dell'integrazione diagnostica e terapeutica nel carcinoma della cervice uterina localmente avanzato (stadi FIGO IIB-III). Ecografia transrettale e TC nella stadiazione e nel controllo dopo terapia. Risultati preliminari.

The diagnosis, staging, therapy and follow-up of advanced epidermoid cervical carcinoma (FIGO stages IIB-III) have been studied in our university since January 1990. By December 1992, a team of specialists including radiologists, radiotherapists, gynecologists and pathologists divided 54 patients into two random treatment groups: group A patients, after systemic chemotherapy (CDP, 2 cycles) and diagnostic reevaluation, underwent radical surgery; group B patients received conventional radiotherapy alone (ERT 45 Gy+IRT or END-RT 20-25 Gy). All patients were examined by means of transrectal US (TRUS) and CT, after clinical examination under sedation, at staging and during the follow-up. The exams were performed periodically for group B patients and after systemic chemotherapy for group A patients. Imaging findings were compared with pathology only in group A. All imaging results were filed. The results confirm some literature data--e.g., 62% diagnostic accuracy for CT and 69% for TRUS, with higher diagnostic accuracy of the latter to evaluate cervical volume and to diagnose local relapses. As for parametrial involvement, both imaging methods tend to understage the early involvement, but only CT tends to overstage the lesions, especially in irradiated patients, due to fibrosclerosis phenomena. TRUS exhibited 69% accuracy, 70% sensitivity and 69% specificity, versus 61%, 62% and 60%, respectively, for CT; clinical examination under sedation had 58%, 60% and 60%, respectively. Both TRUS and CT are faster than endoscopic methods in evaluating vesical and/or rectal involvement. Lymph node metastases at staging, especially those in lumboaortic locations, proved to be unfavorable prognostic signs, as demonstrated by lumboaortic lymph node relapses in 5 group B patients (only 2 of them presented with lymph node metastases at staging; 3 patients had micronodules near the renal vessels), in spite of good local response after radiotherapy. In conclusion, we would like to point out that our team has had an MR unit at its disposal only recently: since the method is considered as the gold standard of imaging, especially in this kind of lesions, the study is still in progress.