Prognostic significance of presence and reduplication of basal lamina in malignant pleural mesothelioma.

The prognosis of patients with malignant pleural mesothelioma (MM) is dependent more on tumor extension and differentiation than on therapeutic effects. Reduplication of the basal lamina (RBL) is an ultrastructural feature of some benign and malignant tumors that has been inversely correlated with aggressiveness and was recently described in MM. To investigate whether RBL is important for predicting the survival of patients with MM, transmission electron microscopy was used to identify the presence of basal lamina or RBL in biopsy specimens obtained by thoracoscopy from 35 patients. Cox's regression analysis was used to study the relation of these ultrastructural features to survival. Better outcomes were found for patients whose tumors expressed either basal lamina (HR 0.48; 95% CI, 0.09-2.47) or RBL (HR 0.38; 95% CI 0.12-1.22) compared with the reference category, where basal lamina or RBL was not found. The expression of basal lamina and RBL is an important novel prognostic factors in MM. HUM PATHOL 31:1341-1345.

Establishment of four new mesothelioma cell lines: characterization by ultrastructural and immunophenotypic analysis.

The aim of this study was to assess the biological characteristics of four new malignant mesothelioma (MM) cell lines. Since simian virus (SV)40 sequences have been recently detected in MM, SV40 large T antigen (Tag) expression was also analysed. MM cell lines were characterized by morphological, ultrastructural and cytogenetic analysis. Expression of Tag and of relevant MM markers was studied by immunocytochemistry, surface antigens by indirect immunofluorescence and immunomodulating cytokines by enzyme-linked immunosorbent assay (ELISA). The four MM cell lines, established from pleural effusions, showed a slow proliferation rate and pleomorphic changes during culture. Cell lines expressed vimentin, cytokeratins 8 and 18, and the mesothelial antigen recognized by HBME-1 monoclonal antibody, but not carcinoembryonic antigen. Surface human leukocyte antigen (HLA)-class I and intercellular adhesion molecule (ICAM)-1 molecules were present on all the cell lines. While HLA class II and CD86 were constitutively undetectable, HLA-class II was present after interferon (IFN)-gamma stimulation. All cell lines displayed abnormal karyotypes with chromosome 6 abnormalities. Transforming growth factor (TGF)-beta2 and interleukin (IL)-6 were constitutively secreted, while tumour necrosis factor (TNF)-alpha was secreted only in response to lipopolysaccharide. Intranuclear Tag was expressed in two cell lines. The persistence of large T antigen with human leukocyte antigen class I and intercellular adhesion molecule-1 positivity may point to large T antigen as a target for cytotoxic T-lymphocyte-based immunotherapy in some malignant mesothelioma patients.

Cyclooxygenase-independent induction of p21WAF-1/cip1, apoptosis and differentiation by L-745,337, a selective PGH synthase-2 inhibitor, and salicylate in HT-29 cells.

In order to dissect out cyclooxygenase-dependent from cyclooxygenase-independent mechanisms in the antiproliferative effects of selective prostaglandin H synthase (PGHS)-2 inhibitors, we compared the effects of L-745,337 (a highly selective PGHS-2 inhibitor) with sodium salicylate (a weak PGHS inhibitor) on prostanoid production, induction of the cyclin-dependent kinase inhibitor p21WAF-1/cip1, mutant p53 (m273-p53) levels, apoptosis and differentiation in human colon adenocarcinoma HT-29 cells. L-745,337 dose-dependently suppressed the cyclooxygenase activity of HT-29 cells (IC50: 0.24 microM). Four-day treatment with L-745,337 caused a concentration-dependent inhibition of cell growth (IC50: 0.9 mM) associated with the induction of p21WAF-1/cip1 and an increase in the proportion of apoptotic nuclei (EC50: 0.1 and 0.34 mM, respectively) while reducing the levels of m273-p53 (IC50: 0.2 mM). Sodium salicylate, at the concentration of 10 mM that did not affect prostanoid formation, caused a 60% reduction of cell growth associated with a 3-fold induction of p21WAF-1/cip1 and a 60% increase in the proportion of apoptotic nuclei. Ultrastructural analysis showed that L-745,337 (0.5 mM) and sodium salicylate (10 mM) caused the induction of a differentiated phenotype. We conclude that high concentrations of L-745,337 and sodium salicylate inhibit colon cancer cell growth by a mechanism unrelated to cyclooxygenase inhibition that may involve p53-independent induction of the tumor suppressor p21WAF-1/cip1.

Effect of chronic hyperoxia on young and old rat carotid body ultrastructure.

Morphologic, physiological, and biochemical changes occur in the carotid body (CB) during postnatal development in relation to physiological requirements. Chronic normobaric hyperoxia attenuates the carotid chemosensory response to hypoxia. During aging there is less of a CB response to hypoxia, which results in a reduced ventilatory adaptation and chemosensory discharge. To test if the oxygen-sensitive mechanism is affected by chronic hyperoxia in an age-dependent fashion, we have studied structural and ultrastructural aspects of young and old rat CBs. Four groups of six male Wistar rats were used. One group of two-month-old rats and another of 25-month-old rats were kept at room air. The other two groups, age matched, were exposed to 98-100% O2, for 60-65 h, in a large Plexiglas chamber. The rats were anesthetized, CBs were fixed in situ with glutaraldehyde (2.5% in phosphate buffer. pH 7.4, 320 mOsm), and were prepared for electron microscopy. Young hyperoxic rats showed focal necrosis in type I cells, along with an increase of endoplasmic reticulum, Golgi apparatus, and of mitochondria volume, with loss of cristae. These changes were less pronounced in the older rat CBs compared with the young rats. In conclusion, hyperoxia seems to affect the oxygen-sensitive mechanism in the carotid body cells, and the reduced effects shown in the old rat CBs suggest an age-related decreased sensitivity to oxygen.

Basal lamina reduplication in malignant epithelioid pleural mesothelioma.

Malignant mesothelioma of the pleura is divided in three morphological variants: epithelioid, sarcomatous, and biphasic. Histological similarities between epithelioid malignant mesothelioma (EMM) and lung adenocarcinoma are responsible for the difficult differential diagnosis. Monoclonal antibodies are useful for distinguishing the two neoplasms through immunohistochemical phenotyping, although many cases require ultrastructural characterization for definitive diagnosis. In this study, transmission electron microscopic observations of EMM were compared with those of peripheral adenocarcinoma of the lung (PAL). More specifically, the morphology of the basal lamina is described in 23 cases of EMM and 12 cases of PAL. Reduplication of the basal lamina (RBL) was found in 11 cases (48%) of EMM and in none of the PAL cases. The same cases were immunostained for type IV collagen and the localization of this basement membrane component corresponded to the areas where basal lamina was observed. Since RBL has been associated with neoplastic differentiation in other tumors, this novel feature in EMM needs to be evaluated in future prognostic studies in malignant mesothelioma of the pleura. Moreover, RBL expression in EMM may be an additional ultrastructural parameter used in the differential diagnosis between EMM and adenocarcinoma.

[Ischemic preconditioning of the myocardium: the role of changes in the permeability of the coronary microcirculation]. / Precondizionamento ischemico del miocardio: ruolo delle alterazioni della permeabilità del microcircolo coronarico.

In isolated working rat hearts we have evaluated the effects of preconditioning on postischemic coronary endothelial permeability. Isolated Wistar male rat hearts were used and subdivided into three groups: Group A, control hearts submitted to 20 min global normothermic ischemia; Group B, hearts subjected, before ischemia, to preconditioning (three phases of 3 min ischemia, each one followed to 2 min Langendorff reperfusion; Group C, hearts submitted to preconditioning and hypertonic reperfusion (by adding 80 mM sucrose to normal perfusion buffer), in order to increase the effects on postischemic interstitial fluid accumulation (osmotic forces balance). A 65 min working heart reperfusion was also performed to assess functional response. We have evaluate the hemodynamic changes (coronary and aortic flows, systolic aortic pressure, work minute), reperfusion arrhythmias, heart weight changes (ww/dw), myocardial enzyme release (creatine phosphokinase and lactic dehydrogenase) and microcirculation permeability changes (FITC.albumin diffusion), as an index of endothelial function. In Group B, a significant reduction of ischemia-reperfusion damage (functional recovery, enzyme release and arrhythmias) was detected with respect to Group A. In Group C this reduction was significantly more evident with respect to groups A and B. In Groups B and C, a significant reduction in myocardial reperfusion (ww/dw: A: 5.9 +/- 0.5, B: 4.9 +/- 1.1, p < 0.02 vs A, C: 4.4 +/- 0.6, p < 0.01 vs A) edema and FITC-albumin diffusion (A: 32.8 +/- 5.9% area; B: 16.3 +/- 6.1% area, p < 0.01 vs A; C: 13.3 +/- 4.5% area, p < 0.01 vs A), especially in perimyocytic space was also observed. Data show that preconditioning may exert a cardioprotective effect by reducing endothelial postischemic functional alterations (vascular permeability) and reperfusion edema. The importance of fluid diffusion within the interstitium in the development of reperfusion damage is supported by better postischemic recovery in Group C, in which an interference on osmotic load was performed.

[Endothelium-protective effects of epinine (N-methyldopamine) in myocardial ischemia-reperfusion in the isolated working rat heart]. / Effetto endotelio-protettivo dell'epinina (N-metildopamina) in corso di ischemia-riperfusione nel cuore isolato e lavorante di ratto.

The effect of cardiac dopaminergic receptors stimulation with epinine (N-methyldopamine) on reperfusion injury was investigated in isolated working rat heart submitted to 15 min of global ischemia. Isolated Wistar rat hearts (n = 75) were used and subdivided into five groups: Group A control hearts, Group B epinine 10 ng/ml, Group C epinine 20 ng/ml, Group D epinine 40 ng/ml, Group E epinine 80 ng/ml. The drug was added to the perfusion buffer at the beginning of experimental procedures. Hemodynamic parameters, heart rhythm (epicardial ECG), heart weight changes, coronary microvascular permeability (FITC-albumin diffusion) and myocytes damage (necrosis enzymes release, immunoperoxidase labeling anti-LDH antibody) were evaluated. After ischemia in groups B and C a significant reduction of functional alterations and myocytes damage was observed with respect to Group A associated with a significant reduction of reperfusion edema (heart weight: Group A +29 +/- 3.5%, Group B +15 +/- 3.8%, Group C 16 +/- 5%, Group D 27 +/- 5%, Group E 33 +/- 4%). At reperfusion time, a significant proarrhythmic effect occurred only in groups D and E. A significant reduction of postischemic endothelial FITC-albumin diffusion was also observed in groups B and C (FITC-albumin diffusion, Group A 32.8 +/- 6% area, Group B 16.33 +/- 5% area, Group C 21.7 +/- 4.5% area, Group D 30 +/- 5% area, Group E 35 +/- 7% area). Our data show that, in isolated working rat heart, the dopaminergic stimulation with low-doses epinine may exert a cardioprotective effect against ischemia-reperfusion damage by modulating endothelial permeability changes and improving coronary microcirculation. The importance of dopaminergic receptors is also suggested by the evidence that at higher doses, when alpha and beta-adrenoceptors stimulation occurs, this cardioprotective effect is significantly reduced or lost.

[A case of surgical ligature of the sacro-iliac joint from Renaissance Central Italy]. / Un caso di legatura chirurgica sacro-iliaca di epoca rinascimentale dall'Italia Centrale.

The paper reports a case of interbone ligature between the sacrum and right ileum using copper wire in an adult woman whose skeleton, dating from approximately 1600-1700 A.D., was exhumed from the Church of Sant'Egidio in Borrello (Chieti). Having described the ligature and the techniques used, the Authors discuss the reasons for the operation. In conclusion, it is suggested that the most plausible hypothesis is that it represent an interbone ligature during the course of autopsy to allow the recomposition of a corpse with a sacro-ileal dislocation. The absence of signs of sacro-ileal dislocation caused by mechanical levers during the course of autopsy leads the Authors to suppose the sacro-ileal rupture occurred while the subject was still alive. The Authors therefore hypothesise that the woman died while giving birth following the laceration of the pubic symphysis and rupture of the right sacro-ileal joint. The hypothesis of a corpse restoration is also suggested.

[The congenital absence of the permanent upper lateral incisors with the persistence of the deciduous canines in a subject of the Iron Age from the Sulmona (AQ) necropolis]. / Assenza congenita degli incisivi laterali superiori permanenti con persistenza dei canini decidui in un soggetto dell'età del Ferro della necropoli di Sulmona (AQ).

The paper reports a case of bilateral agenesia of the permanent upper lateral incisors associated with the persistence of the deciduous eye teeth placed between the first premolars and the permanent canine teeth which, in turn, had emerged in place of the agenetic lateral incisors (situs dentis inversus) in a adult male from the Iron Age whose skeletal remains were exhumed from tomb no. 58 of the Sulmona necropolis (AQ). The authors place the case in the context of agenesia as found both in today's population and in ancient times.

[Contribution to the knowledge of the biology of Cyclostomes: activity of aldolase in Lampetra planeri (Bloch) during its biological cycle]. / Contributo alla conoscenza della biologia dei Ciclostomi: attivita' aldolasica in Lampetra planeri (Bloch) durante il ciclo biologico.

It has been ascertained that in the Lampetra planeri (Bloch) the muscle and liver aldolase level in the last larval period in which lipids are accumulated strongly increases; this may be due to an increased synthesis or diminished degradation of the enzyme.