Genetic changes in pT2 and pT3 prostate cancer detected by comparative genomic hybridization.

Prostate-specific antigen (PSA) screening has led to a remarkable increase in prostate cancer cases undergoing operative therapy. Over half of patients with locally advanced cancer (>or=pT3) develop rising PSA levels (biochemical failure) within 10 years. It is very difficult to predict which patients will progress rapidly to advanced disease following biochemical failure (BF). Therefore, a more useful prognostic factor is needed to suggest the most appropriate therapies for each patient. To determine chromosomal aberrations, we examined 30 patients with stage pT2 or pT3 primary prostate adenocarcinomas and no metastases (pN0M0) by comparative genomic hybridization (CGH). Laser capture microdissection (LCM) was used to gather cancer cells from frozen prostate specimens. Common chromosomal alterations included losses on 2q23-24, 4q26-28, 6q14-22, 8p12-22 and 13q21-31, as well as gains on 1p32-36, 6p21 and 17q21-22. Losses at 8p12-22 and 13q21-31 were observed more frequently in pT3 than pT2 tumors (P<0.05 and P<0.01, respectively). Losses at 8p12-22 were more frequent in tumors with BF (P<0.05), and those at 13q12-21 were more frequent in tumors with Gleason score (GS) 7 or more than lower GS (P<0.05). These findings suggest that losses of 8p12-22 and 13q21-31 are important determinants of prostate cancer progression.

Cardiac rhabdomyomas exhibit a fetal pattern of atrial natriuretic peptide immunoreactivity.

Rhabdomyomas are the most common heart tumors seen in infancy. However, whether they represent hamartomas or true neoplasms derived from cardiomyocytes is still controversial. The fetal pattern of atrial natriuretic peptide (ANP) expression (predominant in the atrial and ventricular subendocardium) becomes altered during the early postnatal period to that typical of the adult (all atrial cardiomyocytes and some cells in the ventricular impulse-conducting system). To better comprehend the nature and origin of cardiac rhabdomyomas, we investigated the immunohistochemical expression of ANP in seven surgically excised ventricular specimens and two necropsy cases of multiple, atrial, and ventricular rhabdomyomas in children aged 1 to 34 days. Immunogold labeling for ANP at the ultrastructural level was also performed on three ventricular tumors. Although all atrial tumors were immunoreactive for ANP, these usually showed a variable number of faintly positive cardiomyocytes, contrasting with the diffuse and intense immunoreactivity of the surrounding atrial myocardium. ANP was detected in the ventricular tumors of five (56%) of the nine cases. The positive ventricular tumor cells predominated in the subendocardium and areas with prominent fibrous tissue, usually around blood vessels. Immunoelectron microscopy of the ventricular tumors demonstrated rare, positive cytoplasmic granules surrounded by membranes, usually located near the nuclei. We conclude that cardiac rhabdomyomas exhibit a fetal pattern of ANP immunoreactivity, which suggests delayed maturation of the tumoral cardiomyocytes, reinforcing the notion that cardiac rhabdomyomas are fetal hamartomas.

Great amount of C.pneumoniae in ruptured plaque vessel segments at autopsy. A comparative study with stable plaques.

A possible relationship between C.pneumoniae (CP) infection, atherosclerosis and acute myocardial infarction is a debated matter. Now we performed the search of CP in histological segments of fatal ruptured plaques and of stable plaques by histochemistry (Macchiavello stain), immunohistochemistry and in situ hybridization techniques. Electron microscopy and confocal laser microscopy techniques were used in two additional cases. The semi-quantification of CP + cells (0-4+) and quantification of lymphocytes demonstrated greater amount of CP + cells and more inflammation in the adventitia of vulnerable plaque vessel segments than of stable ones, larger amount of CP + cells in adventitia than in the plaque and high frequency of CP + cells in all groups studied. This preliminary study strongly suggests a direct pathogenetic involvement of adventitial CP in the rupture of the atheromatous plaque, development of acute myocardial infarction and also in the development of atherosclerosis.

Different microcirculatory and interstitial matrix patterns in idiopathic dilated cardiomyopathy and Chagas' disease: a three dimensional confocal microscopy study.

OBJECTIVE: To analyse the morphological aspects of the extracellular matrix and microcirculation to clarify whether chronic Chagas' cardiopathy (CCC) is an accurate model to study the pathogenesis of idiopathic dilated cardiomyopathy (IDCM). DESIGN: Thick histological myocardial sections were prepared to analyse collagen, and microcirculation was examined during confocal laser and light microscopy. SETTING: The specimens were prepared at the pathology service of the Heart Institute of São Paulo, Brazil. PATIENTS: Nine control hearts, eight IDCM hearts, and 10 CCC hearts were studied after necropsy. MAIN OUTCOME MEASURES: The number of collagen struts per 100x field, the area of fibrosis (%), and the diameters of arterioles and capillaries were measured in each heart to establish outcome. RESULTS: A smaller number (mean (SD)) of collagen struts was seen in the hearts in the IDCM group (9.1 (4.1)) than in the control (22.4 (3.2)) (p < 0.05) or CCC (15.7 (7.4)) (p > 0.05) groups. Fibrosis was greater in the CCC hearts (13.8 (10.5)%) than in the IDCM hearts (5.9 (6.6)%) (p > 0.05). Major increases in arteriole (65.4 (9.9) microm) and capillary (9.9 (1.7) microm) diameters were seen in the CCC hearts but not in the IDCM hearts (arteriole diameter 40.3 (7.9) microm; capillary diameter 7.9 (1.3) microm). CONCLUSIONS: Hearts demonstrating CCC and IDCM present different extracellular and microvessel alterations. This suggests that distinct pathogenic mechanisms are responsible for each condition and that CCC is not an effective model to study IDCM.

[Systematic quantification of the arteries in lung biopsies of patients with congenital heart defects and its contribution to the therapeutic management]. / Quantificação sistemática das artérias em biópsias pulmonares de pacientes com defeitos cardíacos congênitos e sua contribuição para a indicação e abordagem terapêutica.

PURPOSE: To determine the value of the quantitative analysis of lung biopsies from patients with congenital cardiac defects. METHODS: Fourty nine biopsies were examined, from patients: 43 patients increased pulmonary blood flow, 3 with pulmonary atresia and large systemic-to-pulmonary collateral vessels, and 3 with decreased pulmonary flow. The degree of lesion was determined as in Heath-Edwards and of Rabinovitch and col. RESULTS: The Heath and Edwards grade was determined in 41 cases; I in nine; II in 23; III in eight; IV in one; 3 biopsies showed evidence of reduced pulmonary flow and 5 had no signs of vascular disease. The Down patients (7) presented a greater proportion of severe lesions. Quantitative evaluation was obtained in 35 biopsies: 11 had grade B and 24 had grade C. Wall atrophy and dilatation of intraacinar arteries were detected in 7 cases, what suggested the existence of obstructive lesions in proximal vessels, even if not sampled. Medical thickness greater than 2 times the normal were observed in pre-acinar arteries from 14 biopsies. CONCLUSION: The morphometric approach allowed us to detect severe lesions which the qualitative analysis alone would not indicate adequately. In the patients presenting decreased pulmonary flow, morphometry made possible to assess if the degree of arterial wall hypertrophy was compatible with a surgery of atrio-pulmonary anastomosis.

Induction of superoxide dismutases in Photobacterium leiognathi.

We investigated the induction of Cu,Zn-SOD (bacteriocuprein) and Fe-SOD in Photobacterium leiognathi DK-A1 which was isolated from the light organ of the squid, Droteuthis kensaki. The induction of superoxide dismutases depended on the addition of paraquat to the medium. Induction of SOD by paraquat was attributed mostly to the bacteriocuprein by measuring of the activities of both SODs by using densitometry of isoelectrofocusing gel. When paraquat was added to the culture at various times in the early log phase of growth, the most efficient induction of the SODs, which was measured at the time of harvesting the cells (17 hours after inoculation), was observed when paraquat was added at 60 min after the inoculation. Catalase was not significantly induced by the addition of paraquat or increasing of oxygen concentration. We developed an assay of SOD by modification of a cytochrome c-xanthine oxidase method using a computer equipped absorption spectrophotometer.

[Dissolution of cholesterol mixed stones with methyl tertiary butyl ether in vitro].

Methyl tertiary butyl ether (MTBE) has been proposed as a new therapeutic material for the dissolution of pure cholesterol mixed stones. To determine the effect of solvent circulation and fragmentation on dissolution of cholesterol mixed stone in MTBE, we carried out in vitro experiments. The following results were obtained: 1) The addition of 1% EDTA and 0.5% UDCA in MTBE enhanced the dissolution of cholesterol mixed stones (56 +/- 8% of initial weight after 3 hours vs. 48 +/- 5% of initial weight after 3 hours in MTBE alone). 2) Cholesterol mixed stones with circulation of MTBE + 1% EDTA + 0.5% UDCA were dissolved more rapidly than those without. 3) Under in vitro condition, when cholesterol mixed stones were fragmented with URAT-1M stone disintegrator, fragments of stones were dissolved 96% of initial weight in solvent circulation of MTBE + 1% EDTA + 0.5% UDCA after 3-6 hours. In conclusion, these data indicated that both fragmentation of cholesterol mixed stones and solvent circulation of MTBE + 1% EDTA + 0.5% UDCA considerably accelerated dissolution of cholesterol mixed stones in vitro.

Structural analysis of phospho-D-mannan-protein complexes isolated from yeast and mold form cells of Candida albicans NIH A-207 serotype A strain.

The immunochemical properties between phospho-D-mannan-protein complexes of yeast (Y) and mycelial (M) forms of Candida albicans NIH A-207 (serotype A) strain were compared. Hydrolysis of the Y-form complex gave a mixture of beta-(1----2)-linked D-mannooligosaccharides consisting mainly of tri- and tetra-ose, whereas the M-form complex gave preponderantly D-mannose. The antiserum against Y-form cells exhibited a lower reactivity with the M-form than with the Y-form complex, whereas the antiserum to M-form cells could not distinguish significantly between both complexes. Moreover, these acid-modified complexes showed lower antibody-precipitating effect than each corresponding intact complex against antisera of Y- and M-form cells. Digestion of the acid-modified Y- and M-form complexes with the Arthrobacter GJM-1 strain alpha-D-mannosidase yielded 35- and 40-% degradation products, respectively. Acetolysis of each modified complex under mild conditions gave the same D-mannohexaose, beta-D-Manp-(1----2)-beta-D-Manp-(1----2)-alpha-D-Manp -(1----2)-alpha-D-Manp- (1----2)-alpha-D-Manp-(1----2)-D-Man. Because the complexes of Y- and M-form cells of C. albicans NIH B-792 (serotype B) strain did not give any hexaose fraction containing beta-(1----2) linkages, the presence of this hexaose can be regarded as one of the dominant characteristics of the serotype-A specificity of C. albicans spp.

Thin-section and freeze-fracture study of post-mortem changes in dog myocardium.

1. Fragments of dog hearts submitted to 1, 6, 10, 24 and 48 h of autolysis at 20 degrees C were studied with freeze-fracture and thin-section techniques under the transmission electron microscope. 2. The freeze-fracture replicas revealed maximal reduction in the mean number and clustering of intramembrane particles at 6 h post mortem, indicating irreversible cellular damage. However, signs of lethal damage (intramitochondrial amorphous dense bodies) were not observed in thin sections of the same material. 3. The present study indicates that signs of irreversible damage similar to that occurring in in vivo ischemic alterations can be detected earlier by the freeze-fracture technique than by the thin-section technique.

Thin-section and freeze-fracture study of post-mortem chenges in dog myocardium

Fragments of dog hearts submitted to 1,6,10,24 and 48 h of autolysis at 20-C were studied with freeze-fracture and thin-section techniques under the transmission electron microscope. The freeze-fracture replicas revealed maximal reduction in the mean number and clustering of intramembrane particles at 6 h post mortem, indicating irreversible cellular damage. However, sings of lethal damage (intramitochondrial amorphous dense bodies) were not observed in thin sections of the same material. The present study indicates that signs of irreversible damage similar to that occuring in in vivo ischemic alterations can be detected earlier by the freeze-fracture technique than by the thin-section technique

Relationship between luminous fish and symbiosis. I. Comparative studies of lipopolysaccharides isolated from symbiotic luminous bacteria of the luminous marine fish, Physiculus japonicus.

In order to investigate the relationship between host and symbiosis in the luminous marine fish, Physiculus japonicus, the bacterial lipopolysaccharides (LPS) of symbiotic luminous bacteria were compared serologically and electrophoretically. Five symbiotic luminous bacteria (PJ strains) were separately isolated from five individuals of this fish species caught at three points, off the coasts of Chiba, Nakaminato, and Oharai. LPS preparations were made from these bacteria by Westphal's phenol-water method and highly purified by repeated ultracentrifugation. These LPSs contained little or no 2-keto-3-deoxyoctonate and had powerful mitogenic activity. In sodium dodecylsulfate polyacrylamide gel electrophoresis, these PJ-1 to -5 LPSs were separated by their electrophoretic patterns into three groups; the first group included PJ-1 and PJ-4, the second group PJ-2 and PJ-3, and the third group PJ-5 alone. The results agreed with those of the double immunodiffusion test; precipitin lines completely coalesced within each group but not with other groups. In immunoelectrophoresis, one precipitin line was observed between anti PJ-2 LPS serum and PJ-5 LPS but the electrophoretic mobility of PJ-5 LPS was clearly different from that of the PJ-2 LPS group. Furthermore, in a 50% inhibition test with PJ-2 LPS by the passive hemolysis system, the doses of PJ-2 LPS, PJ-3 LPS, and PJ-5 LPS required for 50% inhibition (ID50) in this system were 0.25, 0.25, and 21.6 micrograms/ml for each alkali-treated LPS, respectively, and the ID50's of both PJ-1 LPS and PJ-4 LPS were above 1,000 micrograms/ml. These results indicate that PJ-5 LPS has an antigenic determinant partially in common with LPS from the PJ-2 group but not with LPS from the PJ-1 group and that the symbiotic luminous bacterium PJ-5 is more closely related to the PJ-2 group than to the PJ-1 group. These results show that the species Physiculus japonicus is symbiotically associated with at least three immunologically different strains of luminous marine bacteria in its specialized light organ.

Enhancement of immune responses and possible inhibition of suppressor cells by aclacinomycin A.

Antitumor antibiotics were examined as possible candidates that possess activity which inhibits preferentially suppressor cells in comparison with effector cells. In screening for such compounds among known antibiotics, aclacinomycin was found to augment antibody formation and delayed-type hypersensitivity in mice over a wide concentration range. The addition of aclacinomycin to mouse spleen cell cultures also enhanced antibody formation in vitro. The generation of suppressor cells or the suppressor activity per se in mice immunized with high doses of SRBC was reduced by aclacinomycin. These results suggest that the drug may possibly inhibit suppressor cells selectively. The administration of aclacinomycin at ow doses exhibited antitumor effects on IMC carcinoma; the effect was not dose-dependent.

Antitumor effect of bactobolin and its influence on mouse immune system and hematopoietic cells.

Bactobolin prolonged survival period of mice bearing leukemia L-1210 in various dose schedules. The administration of bactobolin before or at time of immunization with sheep red blood cells (SRBC) did not affect antibody formation and delayed-type hypersensitivity (DTH) to SRBC. The administration after immunization suppressed antibody formation markedly but not DTH response. Bactobolin showed stronger suppressive action on antibody formation in vitro than mitomycin C. Bactobolin did not reduce establishment of tumor immunity which was mediated by T cells and macrophages. Comparing to other antitumor antibiotics which were effective against L-1210, bactobolin did not affect phagocytosis of mouse peritoneal macrophages. It has an extremely low toxicity to mouse spleen cells treated by concanavalin A (Con A) and lipopolysaccharide (LPS). It did not affect colony formation of mouse bone marrow cells in the presence of LPS-induced colony stimulating factor. The administration of bactobolin did not reduce the number of leucocytes in peripheral blood. From these results, the usefulness of bactobolin in the treatment of cancer was discussed.

Effect of bestatin on mouse immune system and experimental murine tumors.

Effect of bestatin on the establishment of delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC) and oxazolone was examined in normal and immunity-impaired mice. Administration of a low dose of bestatin (0.1 approximately 100 microgram/mouse) augmented DTH to SRBC and restored their impaired DTH to oxazolone. The effect of bestatin in the mouse was age-dependent. Bestatin retarded the growth of slow growing solid tumors of Gardner lymphosarcoma and IMC carcinoma and the effect was influenced by the time of the administration and the number of cells inoculated. Bestatin enhanced the antitumor action of the antitumor antibiotics, bleomycin and adriamycin. Bestatin also retarded the induction of skin cancer by 20-methylcholanthrene.