Human gallbladder carcinoma: Role of neurotrophins, MIB-1, CD34 and CA15-3.

Gallbladder carcinoma is the most common biliary tract tumor and the fifth most common gastrointestinal tract cancer .The prognosis of gallbladder carcinoma is poor and less than 5% of the patients are still alive five years postoperatively. Gallbladder specimens were obtained during surgical operations performed in eleven patients for resection of a gallbladder carcinoma, and during five autopsies (control cases selected among patients who died from for other causes, excluding those suffering from biliary or hepatic diseases). Immunohistochemical characterization and distribution of neurotrophins, with their respective receptors, were analyzed. The actual role played by these neurotrophic factors in the general regulation, vascular permeability, algic responsiveness, release of locally active substances and potential tumorigenesis in the gallbladder and biliary ducts compartment remains controversial. Our study revealed an increased immunohistochemical expression of NGF and TrKA in the epithelium and in the epithelial glands of the gallbladder carcinoma together with an evident immunoreactivity for BDNF in the same neoplastic areas. An evident immunoreactivity for NGF, TrKA and BDNF was observed in control specimens of gallbladder obtained during autopsies, whereas a weak or quite absent immunoreactivity was observed in the same specimens for NT4, TrKC and p75. On the contrary an appreciable immunoreactivity for p75 was observed in the specimens harvested from patients with gallbladder carcinoma. We also investigated the expression of some known tumor markers such as MIB-1 (anti Ki-67), CD34 and CA15-3, to identify a possible correlation between the expression of these molecular factors and the prognosis of gallbladder carcinoma. They resulted highly expressed in the stroma (CD34 and CA 15-3) and in the epithelium/epithelial glands (MIB-1) of the neoplastic areas and appeared to be almost absent in the control cases, suggesting that these markers, taken together, could be adopted as a panel of prognostic factors in the evaluation of the gallbladder carcinoma.

Serum CA 15-3 is increased in pulmonary fibrosis.

BACKGROUND AND AIM OF THE WORK: Carbohydrate antigen CA 15-3 is a glycoprotein whose expression, aberrant intracellular localization and changes in glycosylation have been associated with a wide range of cancers. Pulmonary fibrosis represents the final evolution of a chronic inflammation and is defined by the overgrowth of fibroblasts and exaggerated extracellular matrix deposition. The aim of the present study was to evaluate the possible diagnostic role of CA 15-3 in fibrosis in different idiopathic interstitial pneumonias. METHODS: CA 15-3 was measured in serum samples from healthy subjects (n=25) and patients affected with idiopathic pulmonary fibrosis (IPF/UIP) (n=20), sarcoidosis (n=22) at different stages (I, II, and III) and systemic sclerosis (n=25). CA 15-3 protein expression was also evaluated by immunohistochemistry in 21 lung biopsies and in 6 primary lung fibroblasts cell lines. RESULTS: The CA 15-3 serum levels were significantly higher in patients with IPF/UIP and with clinically advanced sarcoidosis (stage III). Serum CA 15-3 levels were slightly increased in patients with systemic sclerosis. No difference was observed between serum CA 15-3 levels in patients with sarcoidosis at stages I and II compared with control subjects. In IPF/UIP and in sarcoidosis at stage III elevated CA 15-3 serum levels significantly correlated with decreased total lung capacity, decreased diffusing capacity of carbon monoxide and high resolution computed tomography findings. Immunohistochemical analysis showed an intense specific CA 15-3 staining in fibroblasts within fibroblastic foci, surrounding sarcoid granulomas and in all cell cultures of lung fibroblasts from IPF/UIP lungs. CONCLUSIONS: Our results indicate that increased CA 15-3 levels are associated with pulmonary interstitial damage, fibroblast activity and progression to fibrosis of the lung. Therefore, CA-15-3 may be considered a sensitive marker useful in the identification of patients with advanced fibrosis and more severe prognosis.

A possible role of BDNF in prostate cancer detection.

Many studies have demonstrated that both normal and malignant prostate cells respond to a variety of growth factors, while several significant differences were found between normal and tumoural cells. The aim of this study was to focus on the localization and distribution of the immuno-reactivity for neurotrophins (NTs) and neurotrophin receptors (NTRs) in normal, hyperplastic and prostate cancer cells, obtained from 40 subjects. We studied samples obtained from 16 prostate cancer (PC, retropubic radical prostatectomy), 20 benign prostatic hyperplasia (BPH, supra-pubic prostatectomy) and normal peripheral prostate tissue from four fresh male cadavers. Samples were examined via immunohistochemical techniques in order to detect the expression of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin 3 (NT3) and their own receptors TrkA, p75, TrkB and TrkC. We observed a high expression of BDNF and TrkB in PC and BPH, though no immuno-reactivity was found for p75. Low expression was reported by other NTs and NTRs in the normal peripheral prostate zone, BPH and PC. These data suggest a possible predictive role for NTs and NTRs, especially for BDNF and TrkB, in the diagnosis and/or management of prostate cancer. The absence of p75 expression confirms its supposed role in apoptotic phenomenon.

Immunohistochemical profile of some neurotransmitters and neurotrophins in the seminiferous tubules of rats treated by lonidamine.

Lonidamine (LND) or [1-(2,4-dichlorobenzyl)-1H-indazole-3-carboxylic acid] is an anticancer and antispermatogenic drug that exerts a large number of effects on tumor cells and germ cells. Sexually mature male Sprague-Dawley rats were housed at 22 degrees C on a 12-h light/12-h dark cycle 1 week before the experiments, with free access to food and water. LND was suspended in 0.5% methylcellulose at a concentration of 10 mg/mL and administered orally at the dose of 10 mL/kg (b.w.) as a single dose. Control rats received an equal amount of vehicle. Testes were removed, fixed for 24 h in 2% glutaraldehyde and 2% paraformaldehyde in 0.1 M sodium phosphate (pH 7.2 at 22 degrees C), rinsed with the same buffer, and stored at room temperature. From each sample, a block of tissue was removed by sectioning through the organ. After dehydration in ethanol at increasing concentrations (70-100%), each block was embedded in paraffin and serial 5 mm thick sections were cut using a rotatory microtome. The immunoreactivity for NTs has been observed in spermatogonia of untreated rats, while the rats treated with LND showed an immunohistochemical localization in all the stages of germinal cells. The generally well-expressed immunoreactivity for the neurotrophins receptors in treated rats observed in our study is presumably attributable to alterations of the receptors' structure and/or expression leading to changes of the activity, affinity, localization or protein interactions that may depend on sensitization of ion channels (induced by LND). Neurotrophins (NTs) appear to be interesting proteins for the modulation of sperm maturation and motility with a prominent role for the nerve growth factor (NGF), that may exert an autocrine or paracrine role. We therefore investigated the location and distribution of immunoreactivity for some neurotransmitters (SP, VIP, CGRP, nNOS, Chat), neurotrophins (NGF, BDNF, NT-3) and their own receptors (TrKA, TrKB, TrKC, p75) in the seminiferous tubules of male rats treated by LND in the light of the literature on this topic.

Expression of neurotransmitters and neurotrophins in human adenoid tissue.

Mucosae-associated lymphoid tissues are richly innervated and the mucosae contain peptidergic nerve endings associated with different types of cells and macrophages. The lymphatic tissue is known to interact with the nervous system and several organs, implicated in the host response to a wide range of stressors, and is also richly innervated. We focussed our attention on the immune organs with particular regard to the human adenoid lymphatic tissues in order to investigate the neuroimmune links and the possible existence of relationships among different neurotransmitters and lymphocytes, macrophages, epithelial cells and nerve fibers by testing the expression of certain neurotransmitters and neurotrophins (NTs) with their own receptors.

Neurotrophins and neurotrophin receptors in human lung cancer.

The expression of neurotrophins (NTs) and related high- and low-affinity receptors was studied in surgical samples of histologically diagnosed human tumors of the lower respiratory tract. The experiment was conducted with 30 non-small cell lung cancer specimens and in eight small cell lung cancer specimens by Western blot analysis and immunohistochemistry to assess expression and distribution of NT and NT receptor proteins in tissues examined. Immunoblots of homogenates from human tumors displayed binding of anti-nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and NT-3 antibodies as well as of anti-tyrosine-specific protein kinase (Trk) A, TrkB, and TrkC receptor antibodies, with similar migration characteristics than those displayed by human beta-NGF and proteins from rat brain. A specific immunoreactivity for NTs and NT receptors was demonstrated in vessel walls, stromal fibroblasts, immune cells, and sometimes within neoplastic cell bodies. Approximately 33% of bronchioloalveolar carcinomas exhibited a strong membrane NGF and TrkA immunoreactivity, whereas 46% adenocarcinomas expressed an intense TrkA immunoreactivity but a weak immunostaining for NGF within tumor cells. Moreover, squamous cell carcinomas developed an intense TrkA immunoreactivity only within stroma surrounding neoplastic cells. A faint BDNF and TrkB immunoreactivity was documented in adenocarcinomas, squamous cell carcinomas, and small cell lung cancers. NT-3 and its corresponding TrkC receptor were found in a small number of squamous cell carcinomas within large-size tumor cells. No expression of low-affinity p75 receptor protein was found in tumor cells. The detection of NTs and NT receptor proteins in tumors of the lower respiratory tract suggests that NTs may be involved in controlling growth and differentiation of human lung cancer and/or influencing tumor behavior.

[Inflammatory pseudotumor of the bladder: etiopathologic and clinical features]. / Pseudotumore infiammatorio vescicale: aspetti etiopatologici e clinici.

The Authors present a case of inflammatory pseudotumour (IPT) of the urinary bladder occurring in a 57-year-old female patient, who was referred to our department with haematuria, stranguria and hypogastric pain. Ultrasonographic, radiological and endoscopic examinations showed a sessile, ulcerated, easily bleeding bladder formation; urinary cytology revealed no atypical transitional cells. Abdomino-pelvic computed tomography analysis showed thickening of the bladder walls and infiltration of the perivesical fat. Histopathologically, the formation was indicated as an inflammatory pseudotumour (IPT) of the bladder. The patient underwent TURB (transurethral resection of the bladder) and was discharged clinically healed on postoperative day 4. A one-year follow up revealed no evidence of recurrence. On the basis of their experience and a thorough review of the literature review, the Authors discuss the clinico-pathological features of IPT of the bladder and the possible factors involved in the malignant transformation of IPT. In conclusion, the benign nature of the lesion is stressed.

Neurotrophins and neurotrophin receptors in human pulmonary arteries.

The localization of neurotrophins (NTs) and NT receptors was analyzed in sections of human extra- and intrapulmonary arteries by Western blot analysis and immunohistochemistry. In extrapulmonary branches of human pulmonary artery, NT and NT receptor immunoreactivity was located in the tunica intima, within endothelium, in the tunica media, within smooth muscle and in the tunica adventitia. In different sized intrapulmonary arteries, NT and NT receptor immunoreactivity was observed primarily in the tunica adventitia. A faint NT and NT receptor immunoreactivity was observed in the tunica media of large-sized branches of intrapulmonary arteries, but not within medium- or small-sized intrapulmonary vessels or in tunica intima of different sized intrapulmonary arteries. These findings suggest that NTs may have a role in the control of vascular responses in the pulmonary system acting as local paracrine or autocrine mediators. The possible relevance of the NT system in human pulmonary vasculature identified in this study is discussed.

Neurotrophin and neurotrophin receptor expression in alveolar macrophages: an immunocytochemical study.

Alveolar macrophages play a crucial role in regulating lung immune responses and in maintaining the integrity of the respiratory tract. Neurotrophins (NTs), besides to their neurotrophic activities, exhibit physiological effects in the immune system. In this study, nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), NT-3 and low- (p75) and high affinity (Trks) NT receptors were investigated by immunocytochemistry in cytospin centrifuged preparations of human alveolar macrophages. Approximately 2.5% alveolar macrophages were immunoreactive for NGF, whereas no macrophages displaying immunoreactivity for BDNF or NT-3 were observed. A 3.5% macrophages displayed immunoreactivity for TrkA-receptor protein, 10% for TrkB-receptor protein (full length isoform), and 2% for TrkC-receptor protein. No low-affinity p75 NT and TrkB[-] truncated isoform receptor immunoreactive macrophages were found. These findings support the hypothesis that NTs and the corresponding receptors may play a role in regulating immunological and functional activity of alveolar macrophages via paracrine/autocrine mechanisms.

[Pseudosarcoma of the urinary bladder. Report of a case]. / Pseudosarcoma della vescica urinaria. Studio di un caso.

A case of myofibroblastic pseudosarcomatous tumor of the bladder occurred in a patient previously submitted to TURB is reported. Aim of this investigation is to demonstrate, on the basis of the literature, the importance of histopathology in the diagnosis of bladder pseudosarcoma, since endoscopical and radiological methods can erroneously show a neoplasm. A 65-year-old male patient with hematuria, submitted to ultrasonography and cystoscopy, revealed a bladder superficial papillary formation. A TURB was performed and the histological study showed a superficial bladder cancer (T1-G1-2). After a second hematuria episode with stranguria a new bladder formation was diagnosed and resected; the histological study revealed a bladder pseudosarcoma. The absolute benignity of the lesion and the slight difference in the symptomatology between urothelial carcinoma and pseudosarcoma are underlined. Furthermore, the importance of histopathology to obtain an accurate diagnosis is stressed. The difficulty of a preoperative diagnosis without histopathological examination is pointed out.

Distribution of p75 and trk-neurotrophin receptor proteins in adult human sympathetic ganglia.

We investigated the expression of immunoreactivity (IR) for low- (p75) and high-affinity (trk proteins) neurotrophin-receptor proteins in adult human paravertebral-sympathetic ganglion neurons. Mouse monoclonal antibodies against the pan-neurotrophin-receptor p75, and rabbit polyclonal antibodies against specific epitopes of the intracytoplasmic domain on trk neurotrophin-receptor proteins were used in fresh unfixed and formaldehyde-fixed paraffin-embedded sympathetic ganglia. All adult human paravertebral-sympathetic neurons displayed trkA neurotrophin-receptor-like protein IR, 10% express trkC neurotrophin-receptor-like protein IR, 37-44% show p75 IR, and no IR was obtained for trkB neurotrophin-receptor-like protein. The intensity of immunostaining was independent of the neuron size. Labelling of non-neuronal tissues, especially blood-vessel walls, was observed for p75, trkA and trkC neurotrophin-receptor proteins. These results indicate that overlapping exists in the expression of p75 and trk neurotrophin-receptor proteins in adult human paravertebral-sympathetic neurons, and suggest that neurotrophins might act on these neurons.

Immunohistochemical localization of neurotrophin receptor proteins in human skin.

The target organs of neurotrophin-dependent sympathetic and sensory neurons, including the skin, synthesize and release neurotrophins, primarily NGF. Neurotrophins undergo retrograde axonal transport, and exert specific function in the perikarya of the responsive neurons. Moreover, evidence exists for an autocrine and/or paracrine function of neurotrophins in the skin. This study analyses the immunohistochemical localization of low (gp75) and high-affinity (gp140 trkA, gp145trkB and gp145trkC) neurotrophin receptor proteins in the human glabrous skin. We consider that the expression of neurotrophin receptors may be indicative of neurotrophin activity. Specific gp75 and gp140trkA-like immunoreactivity (IR) were observed highly co-localized in (1) epidermis, primarily in the basal keratinocytes; 2) sweat glands; (3) blood vessel walls, mainly in the muscular layer; (4) Schwann and perineurial cells of nerve trunks; (5) periaxonic cells forming sensory nerve formations (Meissner's and Pacini's corpuscles); (6) large axons of nerve bundles and of sensory corpuscles; gp145trkB-like and gp145trkC-like were found labelling nerve fibers and sensory nerve formations, as well as blood vessels and sweat glands, but not epidermic cells. The results suggest that, in addition to the well known neurotrophic functions, neurotrophins may also regulate unknown functions in non-nervous cutaneous cells, which are targets for neurotrophin-dependent sympathetic and sensory neurons.

Effect of vasoactive intestinal polypeptide on the cyclic adenosine monophosphate generating system in rat kidney glomeruli and cortical tubules.

The effect of in vitro addition of vasoactive intestinal polypeptide (VIP) on the 3'-5'-cyclic adenosine monophosphate (cAMP) generating system in rat kidney glomeruli and cortical tubules was studied. VIP did not stimulate cAMP accumulation in glomeruli; but VIP did stimulate, specifically and in a dose-dependent manner, cAMP concentrations in tubular membranes with the addition of GTP. These results are consistent with the existence of a functionally active VIP receptor coupled to adenylate cyclase in rat kidney cortical tubules.

Histochemical studies of some hepatic enzyme activities in rat during extrahepatic common bile duct obstruction.

Studies on some enzyme activities as measured histochemically in the ligated common bile duct rat liver are reported. They show principally: an increased GGT reaction in the neoductular cells; a diffusion of the LAP reaction to the neoductuli; an immediate increase and diffusion of the alkaline phosphatase reaction; an increase of G-6-PA and AMPA reactions; an increase, during the first week, of the Phyla activity and a contemporary decrease of the UDPG-GS reaction. No significant changes have been observed in other enzyme activities. By means of some dehydrogenase reactions, morphological evidence that the hepatocytes may be involved in neoductular cells formation was obtained.