Tumor and peritumoral adipose tissue crosstalk: De-differentiated adipocytes influence spread of colon carcinoma cells.

Colorectal cancer is the second leading cause of cancer and often has a fatal course. There are many studies in the literature that have described a close functional relationship between the tumor mass and surrounding tissue, or tumor stroma, which is affected by the continuous metabolic exchange that occurs at the interface between tumor and tissues in contact with it. There is much evidence that the presence of adipose tissue in stroma plays a fundamental role in modulating the tumor microenvironment and promote tumor development, growth, and angiogenesis due to its endocrine characteristics. In this analysis, we have studied the alterations of adipose tissue surrounding colorectal tumors with MRI and optical imaging in vivo techniques to monitor tumor progression and also performed histological and molecular analysis. We detected differences in the principal adipose markers expressed by adipocytes residing around the rectal colon and observed that peritumoral adipose tissue is exposed to a mesenchymal transition process that leads to the acquisition of a less differentiated phenotype of adipocyte that represents the main cellular type present in tumor stroma. The mesenchymal transition correlated with the acquisition of more aggressive tumor phenotype and could represent a valid target for tumor therapy.

Pelvic floor rehabilitation for defecation disorders.

Pelvic floor rehabilitation is frequently recommended for defecation disorders, in both constipation and fecal incontinence. However, the lack of patient selection, together with the variety of rehabilitation methods and protocols, often jeopardize the results of this approach, causing difficulty in evaluating outcomes and addressing proper management, and above all, in obtaining scientific evidence for the efficacy of these methods for specific indications. The authors represent different gastroenterological and surgical scientific societies in Italy, and their aim was to identify the indications and agree on treatment protocols for pelvic floor rehabilitation of patients with defecation disorders. This was achieved by means of a modified Delphi method, utilizing a working team (10 members) which developed the statements and a consensus group (15 members, different from the previous ones) which voted twice also suggesting modifications of the statements.

Mutant FGFR3 associated with SADDAN disease causes cytoskeleton disorganization through PLCγ1/Src-mediated paxillin hyperphosphorylation.

K650M/E substitutions in the Fibroblast growth factor receptor 3 (FGFR3) are associated with Severe Achondroplasia with Developmental Delay and Acanthosis Nigricans (SADDAN) and Thanatophoric Dysplasia type II (TDII), respectively. Both SADDAN and TDII present with affected endochondral ossification marked by impaired chondrocyte functions and growth plate disorganization. In vitro, K650M/E substitutions confer FGFR3 constitutive kinase activity leading to impaired biosynthesis and accumulation of immature receptors in endoplasmic reticulum (ER)/Golgi. From those compartments, both SADDAN-FGFR3 and TDII-FGFR3 receptors engender uncontrolled signalling, activating PLCγ1, signal transducer and activator of transcription 1, 3 and 5 (STAT1/3/5) and ERK1/2 effectors. Here, we investigated the impact of SADDAN-FGFR3 and TDII-FGFR3 signalling on cytoskeletal organization. We report that SADDAN-FGFR3, but not TDII-FGFR3, affects F-actin organization by inducing tyrosine hyperphosphorylation of paxillin, a key regulator of focal adhesions and actin dynamics. Paxillin phosphorylation was upregulated at tyrosine 118, a functional target of Src and FAK kinases. By using Src-deficient cells and a Src kinase inhibitor, we established a role played by Src activation in paxillin hyperphosphorylation. Moreover, we found that SADDAN-FGFR3 induced FAK phosphorylation at tyrosines 576/577, suggesting its involvement as a Src co-activator in paxillin phosphorylation. Interestingly, paxillin hyperphosphorylation by SADDAN-FGFR3 caused paxillin mislocalization and partial co-localization with the mutant receptor. Finally, the SADDAN-FGFR3 double mutant unable to bind PLCγ1 failed to promote paxillin hyperphosphorylation, pointing to PLCγ1 as an early player in mediating paxillin alterations. Overall, our findings contribute to elucidate the molecular mechanism leading to cell dysfunctions caused by SADDAN-FGFR3 signalling.

Expression of µ-protocadherin is negatively regulated by the activation of the ß-catenin signaling pathway in normal and cancer colorectal enterocytes.

Mu-protocadherin (MUCDHL) is an adhesion molecule predominantly expressed by colorectal epithelial cells which is markedly downregulated upon malignant transformation. Notably, treatment of colorectal cancer (CRC) cells with mesalazine lead to increased expression of MUCDHL, and is associated with sequestration of ß-catenin on the plasma membrane and inhibition of its transcriptional activity. To better characterize the causal relationship between ß-catenin and MUCDHL expression, we performed various experiments in which CRC cell lines and normal colonic organoids were subjected to culture conditions inhibiting (FH535 treatment, transcription factor 7-like 2 siRNA inactivation, Wnt withdrawal) or stimulating (LiCl treatment) ß-catenin activity. We show here that expression of MUCDHL is negatively regulated by functional activation of the ß-catenin signaling pathway. This finding was observed in cell culture systems representing conditions of physiological stimulation and upon constitutive activation of ß-catenin in CRC. The ability of MUCDHL to sequester and inhibit ß-catenin appears to provide a positive feedback enforcing the effect of ß-catenin inhibitors rather than serving as the primary mechanism responsible for ß-catenin inhibition. Moreover, MUCDHL might have a role as biomarker in the development of CRC chemoprevention drugs endowed with ß-catenin inhibitory activity.

Removal of Congo red dye from aqueous solutions using a halloysite-magnetite-based composite.

Adsorption has been considered as one of the most effective methods to remove dyes from aqueous solutions due to its ease of operation, high efficiency and wide adaptability. In view of all these aspects, this study aimed to evaluate the adsorption capacity of a halloysite-magnetite-based composite in the removal of Congo red dye from aqueous solutions. The effects of stirring rate, pH, initial dye concentration and contact time were investigated. The results revealed that the adsorption kinetics followed the pseudo-second-order model, and equilibrium was well represented by the Brunauer-Emmett-Teller isotherm. The thermodynamic data showed that dye adsorption onto the composite was spontaneous and endothermic and occurred by physisorption. Finally, the composite could also be regenerated at least four times by calcination and was shown to be a promising adsorbent for the removal of this dye.

Mesalazine inhibits the beta-catenin signalling pathway acting through the upregulation of mu-protocadherin gene in colo-rectal cancer cells.

BACKGROUND: Several reports indicate that mesalazine (5-aminosalicylic acid, 5-ASA) is a promising candidate for the chemoprevention of colo-rectal cancer because of its ability to reach the purpose avoiding the unwanted side effects usually associated with prolonged administration of nonsteroidal anti-inflammatory drugs. This activity of 5-ASA is probably the consequence of a number of effects determined on colo-rectal cancer cells, consisting of reduced proliferation, increased apoptosis and activation of cell cycle checkpoints and DNA repair processes. A recent observation has suggested that inhibition of beta-catenin signalling could induce these cellular effects. AIM: To characterize better the capacity of 5-ASA to inhibit the beta-catenin signalling pathway. METHODS: Genes belonging to the beta-catenin signalling pathway were analysed in colo-rectal cancer cell lines treated with 5-ASA using a combination of laboratory assays that are able to detect their phenotypic expression and functional activity. RESULTS: The results obtained indicated that 5-ASA induces the expression of a protein called mu-protocadherin that belongs to the cadherin superfamily and is able to sequester beta-catenin on the plasmatic membrane of treated cells hampering its function. CONCLUSION: These findings suggest that mu-protocadherin might be employed as a biological marker to monitor the chemopreventive efficacy of 5-ASA.

Non-cardiac chest pain: a real clinical problem.

Non-cardiac chest pain is a frequent clinical problem. Between 10 and 50% of patients with anginal pain who are referred for arteriography are found to have normal coronary arteries. An oesophageal source of non-cardiac chest pain is reported in up to 60% of cases, most of which are attributable to gastro-oesophageal reflux disease. The exclusion of heart disease and the identification of an oesophageal origin of the pain may require an extensive work-up. The outcome in patients with non-cardiac chest pain is influenced by both the underlying diagnosis and the patient's perception of his or her symptom.

Interictal abnormalities of regional cerebral blood flow in migraine with and without aura.

Some controversial issues exist whether regional cerebral blood flow (rCBF) changes are present both in migraine with and without aura during the interictal period. For this reason we have studied rCBF characteristics in migraine patients when headache-free. rCBF examinations were performed by the 133Xe inhalation method on 39 normal subjects (24 aged 45 or less and 15 older than 45), on 10 migraine patients with (A+) and on 10 without (A-) aura. The values of each patient were compared with the age-matched control population mean by a computer-assisted mapping system that allows statistical analysis in real time. To compare inter-individual variability 10 subjects, out of 39 normals, constituted an age-, sex- and CO2-matched control group (C). 8 A+ patients and 7 A- showed significant alterations of CBF in comparison with the age-matched control population. The analysis between the age-, sex- and CO2 matched groups showed significant differences of the inter-hemispheric (F = 6.669, p = 0.004) and of the frontal (F = 7.480 p = 0.0008) asymmetries. These data show that in the headache-free period a derangement of the cerebral perfusion is present in both migraine with and without aura, suggesting they are due to the same disease process. Furthermore they show the usefulness of a computer-assisted mapping system, suitable for clinical use, in discovering small alterations in cerebral perfusion.

Influence of erythrocyte aggregability and plasma fibrinogen concentration on CBF with aging.

The influence of the rheological properties of the blood on cerebral perfusion is still unresolved. Data on normal subjects are lacking and difficulties arise regarding the effect of blood viscosity owing to its close relationship with hematocrit. For these reasons we have studied the relationship between two rheological hematocrit-independent parameters and CBF in normal subjects of various ages. 36 normal volunteers, aged 20-74, free from risk factors, have been studied. CBF was measured by the Xenon inhalation method. Erythrocyte aggregability was expressed as Mean Erythrocyte Aggregation Index (MEA). Plasma fibrinogen concentration was evaluated by the coagulative method in 26 subjects. No correlation was found between CBF and MEA or fibrinogen in the subjects under the age of 45. A significant negative correlation was found between CBF and MEA (p = 0.015) and between CBF and fibrinogen (p = 0.011) in the subjects over 45. These data show that cerebral perfusion is influenced by the rheological properties of the microcirulation only with aging. We suggest that a "rheological autoregulation" exists and that it works properly in youth, only to be lost with physiological aging. This finding can be of significance in the pathogenesis of cerebrovascular disease processes in humans.

Ambulatory 24-hour oesophageal pH monitoring in normal subjects: a multicentre study in Italy. G.I.S.M.A.D. GOR Study Group.

A multicentre study was performed in Italy in order to establish normal values for acid gastro-oesophageal reflux. In 73 healthy subjects 24-hour oesophageal pH measurements were carried out under standardized conditions. The hourly number of reflux episodes and oesophageal acid exposure was assessed and analyzed by means of a computerized system. The distribution frequency of both parameters was skewed to the left, thus exhibiting a non-normal pattern. Acid reflux was lowest during the night and highest after meals. The oesophageal acid exposure and the number of reflux episodes were correlated both over 24 hours and in the subsets studied. A good agreement was found with data from previous reports. Our findings should be helpful in establishing a data base for physiological gastro-oesophageal reflux.

CBF side-to-side asymmetries in stenosis-occlusion of internal carotid artery. Relevance of CT findings and collateral supply.

Cerebral blood flow (CBF) was studied at rest by 133-Xenon inhalation method in 15 normal subjects, in 10 patients with cerebral infarction and normal angiograms of major cerebral arteries and in 28 patients with unilateral stenosis-occlusion of the internal carotid artery (ICA), with or without cerebral infarction. All the normals and 20 patients with ICA stenosis-occlusion were tested again after cerebral vasodilatation induced by an intravenous bolus of acetazolamide. At rest the patients with cerebral infarction, irrespective of whether ICa stenosis-occlusion was present or not, showed abnormal side-to-side CBF asymmetry. After cerebral vasodilation variations in side-to-side asymmetry were shown to depend on the inefficiency of the collaterals and not on the degree of ICA obstruction or on the presence of cerebral infarction. These data indicate that at rest side-to-side CBF distribution is influenced more by the presence of an ischemic zone than by a ICA stenosis-occlusion and that under these circumstances the hemodynamic effect of the vascular stenosis cannot be assessed. After cerebral vasodilatation a subset of patients--irrespective of the degree of ICA obstruction and of whether or not cerebral infarction is present--with true cerebrovascular insufficiency can be isolated.

[Esophageal and gastric acidity determination: its value and limitations]. / La pH-metria esofago-gastrica: valori e limiti.

Combined esophago-gastric pH measurements has been recently employed to better define gastroesophageal reflux and to simultaneously evaluate esophageal and gastric acidity. We studied 21 patients with esophagitis and 11 controls. Gastric acidity was rather similar in the two groups. A significant difference (p less than 0.05) between controls and patients was found in the postprandial period only. The analysis of the buffering effect of a standardized meal did not show any difference between the two groups. When subdividing the esophagitis patients according to the outcome of medical treatment we could find a more prolonged (p less than 0.05) postcibal recovery phase. Gastric alkalinizations were observed mostly during the night and did not differ in the two groups. As regards gastro-esophageal reflux we evaluated both acid, mixed and alkaline episodes. Acid reflux was the most frequent event, while mixed reflux was less frequently observed and equally distributed in the two groups. As regards alkaline episodes they were absent in controls and occurred very rarely in the patients. In conclusion esophagogastric pH-metry gives us, with a single test, informations about gastric acidity and the characteristics of gastro-esophageal reflux. On the other hand its clinical relevance needs further evaluations.

Long-term treatment of reflux oesophagitis.

Reflux oesophagitis is a chronic recurrent disease with high tendency towards relapse after medical healing. It has been calculated that after 6 months a symptomatic relapse may occur in about 45% of patients, while a recurrence of mucosal lesions ranges between 20 to 70%. The most important adverse factors affecting the likelihood of recurrence are: a) the daytime symptoms at time of healing and b) an impairment of both oesophageal body motility and LOS tonic and phasic activity. The available data show that the maintenance treatment with anti-H2 at standard doses does not affect the spontaneous recurrence of the disease. Better results could be achieved with higher dosage, possibly tailored to the physiopathological characteristics of the patients.

Acid gastroesophageal reflux and symptom occurrence. Analysis of some factors influencing their association.

In 17 patients with esophagitis (degree I = erythema, N = 10; degree II = erosions, N = 7) esophageal pH was measured at 5 and 10 cm above the esophagogastric junction to assess whether the extension of acid reflux and the severity of the mucosal lesions could influence the association between reflux and symptoms. A minority of the refluxes were related with symptoms (4.0% and 7.7% in degree I, 14.1% and 12.0% in degree II at 5 and 10 cm respectively), whereas 65.4% and 100% of the symptoms were related with reflux in degrees I and II, respectively, with 76.5% and 35.7% occurring during refluxes reaching the proximal recording site. A relationship of symptoms with reflux is shown, particularly in erosive disease. Some reflux characteristics (extension, duration, acidity) seem to influence symptom occurrence mainly in mild esophagitis; however, more than 85% of the acid reflux episodes are symptom-free, regardless of the severity of the mucosal injury.

Oesophageal function before, during, and after healing of erosive oesophagitis.

In order to investigate the relationship between oesophageal motor abnormalities and oesophagitis, we carried out four hour studies of oesophageal motility and 24 hour pH measurements in fasting and fed conditions in eight patients before, during (pH only), and after medical healing of erosive oesophagitis. Gastrooesophageal acid reflux decreased (ns) during the treatment, but tended to return to basal values at the end. Oesophageal body motility was unchanged after healing, while the lower oesophageal sphincter basal tone was significantly increased at the end of the study in the postcibal period. The results suggest that the impairment of the sphincter tone in reflux oesophagitis is secondary to the presence of the oesophageal lesions. Macroscopic healing is not paralleled by improved major pathogenic factors of the disease, however--that is, acid reflux and oesophageal body motility.

Oesophageal motor events at the occurrence of acid reflux and during endogenous acid exposure in healthy subjects and in patients with oesophagitis.

In order to assess the oesophageal motor events associated with the occurrence of gastro-oesophageal acid reflux and those during endogenous acid exposure, we studied six healthy subjects and nine patients with symptoms and lesions of reflux oesophagitis. In the case of each subject simultaneous pressure and pH measurements of the distal oesophagus were taken both in fasting conditions and after a standardised balanced meal. Reflux episodes occurred in the absence of a lower oesophageal sphincter relaxation (34.3%, 17.7%) or in the presence of a relaxation associated (34.2%, 30.8%) or unassociated (31.5%, 51.5%) with a peristaltic sequence, in the controls and patients respectively. No significant differences were found between the two groups. During endogenous acid perfusion the distal oesophagus showed mainly a peristaltic motor activity, but the mean hourly number of peristaltic sequences was significantly lower in the patients than in the controls (33.5 +/- 27.2 vs 81.5 +/- 3.2, p less than 0.01). Our results show that the mechanisms of reflux are similar in controls and in patients, while the peristaltic frequency is reduced in the latter. This motor failure may be responsible for the increased reflux duration observed in patients with oesophagitis.

M-mode subxiphoid echocardiography in assessing pulmonary hypertension. Its usefulness in chronic obstructive pulmonary disease.

12 patients (9 males, 3 females; aged 35-63 years, mean 55) with chronic obstructive pulmonary disease (COPD) were studied in order to determine the correlation between the values of pulmonary arterial pressure and the M-mode echographic size of the right pulmonary artery (RPA) from the subxiphoid approach. Both the largest (systolic) and smallest (diastolic) size of the vessel were considered, and both were corrected for body surface area ('index size' - PA/BSA). The reliability of this echographic size is supported by the evidence that the tract of the pulmonary artery considered in this study is perpendicular to the ultrasonic beam. Both the systolic and diastolic size of RPA correlated with the systolic, diastolic, and mean pulmonary arterial pressure, detected by catheterization. Measurements of RPA size were made from the suprasternal approach in 8 patients out of 12. The size of RPA from the subxiphoid approach was also estimated in 21 normal subjects, serving as controls. Measurements of RPA size from the suprasternal approach were made in 12 of these subjects. Both the systolic and diastolic size of RPA from the subxiphoid approach correlated with the systolic and diastolic size of the suprasternal view in patients and in normal subjects. Subxiphoid M-mode echocardiography seems to be a valuable noninvasive technique in assessment of pulmonary hypertension, particularly in patients affected by COPD.

Oesophageal function after oral ranitidine: an acute double blind study in normal subjects.

The effect of oral ranitidine on oesophageal peristalsis, LOS basal pressure and gastro-oesophageal acid reflux, was investigated in 6 healthy men in a double-blind randomized study. Simultaneous manometry and pH measurements were performed twice in each volunteer during a five hour study period which included the administration of a standard meal. Ranitidine did not affect the motor parameters studied (amplitude, duration and velocity of the peristaltic waves and LOS basal tone), whereas it almost abolished acid gastroesophageal reflux. Our results show that ranitidine, like cimetidine, does not alter the motor function of the oesophagus, while it virtually abolishes acid gastro-oesophageal reflux in normal man.