Immunohistochemical localization of metallothionein in hepatocellular carcinoma: preferential expression in non-cancerous cirrhotic nodules

Metallothionein (MT), an oncofocal gene product was strongly expressed in 35%-95% of hepatocytes in hepatocellular carcinoma (HCC) and MT-positive hepatocytes were localized mainly in the non-cancerous cirrhotic nodules but not in malignant hepatocytes. On the other hand, <10% hepatocytes showed weak staining for MT in chronic hepatitis and cirrhosis of liver. Strong expressions of MT in non-cancerous cirrhotic nodule in HCC and low expressions in liver cirrhosis without HCC indicate a relationship between malignant transformation of hepatocytes and the expression of MT.     

Heat-induced membrane damage combined with adriamycin on prostate carcinoma PC-3 cells: correlation of cytotoxicity, permeability and P-glycoprotein or metallothionein expression

Sodium thiopental, as other amphiphilic molecules, interacts with the membrane by inserting into the lipid bilayer and causing alterations of the membrane properties such as curvature and hypotonic lysis resistance. But can it modify the mechanical properties of the membrane? In the present work it was observed that sodium thiopental affected the membrane rheological properties by improving erythrocyte deformability; this effect resulted from a reduction of both the elastic modulus and surface viscosity. In erythrocytes devoid of sialic acid after treatment with neuraminidase, sodium thiopental membrane concentration was significantly higher than in normal cells, suggesting that drug access to the lipid bilayer be facilitated by the absence of the steric and electrostatic barrier of the glycocalyx negative charges. From a rheological point of view, desialated and normal cells showed the same response to the anesthetic as regards elastic modulus but in opposite direction if surface viscosity was considered. This finding supports the hypothesis that sodium thiopental molecules enter the bilayer of desialated cells in a higher proportion, as compared to the normal erythrocyte, promoting a disorganization that results in a greater inner friction. The changes in the rheological parameters, triggered by sodium thiopental, could be attributed to the bilayer contribution to the membrane mechanical properties, either directly or through interaction between the bilayer and the cytoskeleton.     

p53 and c-jun expression in urinary bladder transitional cell carcinoma: correlation with proliferating cell nuclear antigen (PCNA) histological grade and clinical stage

OBJECTIVE: To investigate p53 and c-jun oncoproteins and proliferating cell nuclear antigen (PCNA) in transitional cell urinary bladder carcinomas (TCCs) and to determine their relationships to tumour grade, stage and survival. MATERIALS AND METHODS: The expression of p53, c-jun and PCNA was studied using immunohistochemistry in formalin-fixed, paraffin-embedded tissues in a series of 110 TCCs. RESULTS: 58% of our cases were positive for p53 and 88% for c-jun. A statistically very significant correlation (p < 0.0001) was observed between p53 and c-jun (r = 0.781), p53 and PCNA (r = 0.772), c-jun and PCNA (r = 0.831) as well as between each of the two oncoproteins and the histological grade and clinical stage (p < 0.001). There was no correlation of either p53, PCNA or c-jun with clinical outcome in terms of patients survival. CONCLUSION: p53 and c-jun proteins' overexpression are strongly related to rapid tumour cell proliferation and hence with aggressive growth in urinary bladder TCC. PCNA score remains an important prognostic index in transitional cell carcinoma of the bladder.     

Immunohistochemical study of desmosomes in oral squamous cell carcinoma: correlation with cytokeratin and E-cadherin staining, and with tumour behaviour

Reduction or loss of the intercellular junctions known as desmosomes may contribute to the invasive and metastatic behaviour of various carcinomas. Previous studies have shown that metastasis of oral squamous cell carcinomas of the head and neck correlates with a reduction in immunohistochemical staining for desmoplakin and desmoglein at the invasion front. The primary aim of the present study was to extend these observations to include a third component of desmosomes, the glycoprotein desmocollin. An additional aim was to determine whether the differentiation status of tumours is reflected in their staining for cytokeratins 1, 13, and 19, and, if so, whether these parameters correlate with desmosomal staining and/or metastasis. The study included 54 primary tumours of which 28 showed lymph node metastases. The results of this investigation show that tumours can be divided into three groups according to whether they have lost staining for no, one or more than one desmosomal component. A statistically significant correlation was found between the number of desmosomal components lost and metastasis. Tumours could also be divided into five groups according to their staining for different combinations of cytokeratins. Furthermore, differentiation status as indicated both histologically and by cytokeratin staining correlated with reduced desmosomal staining and metastasis. Tumours were also examined for intensity of staining for the adhesion molecule E-cadherin. Reduction in E-cadherin staining was correlated with mode of invasion and with reduction in desmosomal staining, but not with poor differentiation as indicated by cytokeratin staining. The results of this extensive study reinforce the view that adhesive junctions and adhesion molecules contribute to the suppression of tumour invasion and metastasis.     

Regulation of metallothionein expression in human amnion epithelial and mesenchymal cells

OBJECTIVE: In this study the potential for metallothionein expression in amnion epithelial and mesenchymal cells in response to cadmium was evaluated. STUDY DESIGN: Levels of metallothionein messenger ribonucleic acid were evaluated in freshly separated amnion epithelial and mesenchymal cells and in amnion cells in culture. RESULTS: The levels of metallothionein messenger ribonucleic acid in human amnion mesenchymal cells freshly isolated after delivery of term pregnancies were greater than those in epithelial cells of the same tissue. The levels in mesenchymal cells in monolayer culture at confluence also were greater than those in confluent epithelial cells propagated from the same tissue. In response to treatment with cadmium (100 nmol/L to 50 micromol/L), which is inhaled in cigarette smoke, the levels of metallothionein messenger ribonucleic acid in both cell types increased markedly in a dose-dependent manner, but the level was greater in epithelial cells at all concentrations of cadmium chloride tested. With cadmium chloride (10 micromol/L), the level of metallothionein messenger ribonucleic acid increased by as much as 1000-fold in epithelial cells and 10-fold in mesenchymal cells compared with untreated (control) cells. Dexamethasone and tetradecanoyl phorbol acetate also acted to increase the levels in amnion epithelial and mesenchymal cells but not nearly to the levels effected by cadmium. CONCLUSION: These findings are indicative that metallothionein expression in amnion epithelial cells is exquisitely sensitive to cadmium in concentrations similar to those in amniotic fluid of pregnancies of women who smoke cigarettes. We hypothesize that increased levels of metallothionein in amniotic fluid and amnion epithelial cells will bind and thereby may limit the availability of copper to the Cu++-dependent enzyme lysyl oxidase in mesenchymal cells and thereby impair the cross-linking of interstitial collagens, which is effected by this enzyme.     

Analysis of K-ras gene mutations in lung carcinomas: correlation with gender, histological subtypes, and clinical outcome

The vertebrate Hox genes have been shown to confer regional identity along the anteroposterior axis of the developing embryo, especially within the central nervous system (CNS) and the paraxial mesoderm. The notochord has been shown to play vital roles in patterning adjacent tissues along both the dorsoventral and mediolateral axes. However, the notochord's role in imparting anteroposterior information to adjacent structures is less well understood, especially as the notochord shows no morphological distinctions along the anteroposterior axis and is not generally described as a segmental or compartmentalized structure. Here we report that four zebrafish hox genes: hoxb1, hoxb5, hoxc6 and hoxc8 are regionally expressed along the anteroposterior extent of the developing notochord. Notochord expression for each gene is transient, but maintains a definite, gene-specific anterior limit throughout its duration. The hox gene expression in the zebrafish notochord is spatially colinear with those genes lying most 3' in the hox clusters having the most anterior limits. The expression patterns of these hox cluster genes in the zebrafish are the most direct molecular evidence for a system of anteroposterior regionalization of the notochord in any vertebrate studied to date.     

Glycosphingolipid component profiles of human gliomas correlate with histological tumour types: analysis of inter-individual and tumour-regional distribution

Three types of glycosphingolipid (GSL) component profiles have been established for human intracranial gliomas. GSL-type I shows only Glac- and lacto-series-sialoglycolipids. Type II consist of Glac- and Gtri-gangliosides, whereas only GSL-type III contains sulphatide and, as a major neutral glycolipid, galactocerebroside, besides gangliosides of the Glac-, Gtri-, and Gtet-families. Whole gliomas of malignancy grading I/II, III and IV, display GSL-Types III, II, and I, respectively. Thus, the GSL component distribution of the samples taken after surgery from three individual gliomas and two biopsies correlate closely with the general diagnosis of these tumours. Arthrobacter ureafaciens sialidase was used for the characterization of gangliosides. GSL-type analysis of multiple regional samples, taken from necropsy and biopsy, were determined by microanalysis of microscopic cryostat section, and shown to be in good agreement with their histology. The results validate the relevance of tumour ganglioside analysis for the characterization and diagnosis of gliomas.     

Protection against pulmonary blastomycosis: correlation with cellular and humoral immunity in mice after subcutaneous nonlethal infection

A model of pulmonary blastomycosis in the mouse, in which the portal of entry is the same as natural human infection, was used to study resistance to challenge after subcutaneous infection. One week after subcutaneous infection, mice were partially resistant to pulmonary challenge, and mice challenged two weeks after infection were resistant. Measurement of cellular and humoral immune responses to Blastomyces dermatitidis antigens after subcutaneous infection showed the following. (i) Delayed-type hypersensitivity appeared 1 week after infection, and responses increased for 3 weeks thereafter. (ii) Proliferative responses in vitro appeared in spleen cells at 1 week and in contralateral lymph node cells at 3 weeks, (iii) Serum antibody, detected by an enzyme-linked immunosorbent assay, appeared 1 week after infection and then increased in titer. (iv) Peritoneal macrophages were activated to inhibit replication of B. dermatitidis in vitro by the first week after infection. Prior subcutaneous infection also resulted in rapid clearing of a second subcutaneous challenge, as well as resistance to a lethal intraperitoneal challenge. This resistance was associated with the development of cell-mediated and humoral immune responses. These data provide a chronological framework for selective transfer experiments.     

Immunohistochemical analysis of metallothionein in astrocytic tumors in relation to tumor grade, proliferative potential, and survival

BACKGROUND: Metallothionein (MT) is the name for a family of predominantly intracellular protein thiol compounds involved in anticancer drug resistance. For certain tumors, MT is related to grade of tumor malignancy and prognosis. The authors evaluated the expression of MT in 114 astrocytic tumors in relation to the proliferative potential of tumors and the survival of patients. METHODS: Paraffin embedded tissue sections were stained with monoclonal anti-metallothionein and MIB-1 Ki-67 antibodies by avidin-biotin complex immunohistochemistry. RESULTS: MT expression was observed in 2 of 6 pilocytic astrocytomas, in 10 of 24 Grade 2 astrocytomas, in 16 of 25 anaplastic astrocytomas, and in 47 of 59 glioblastomas. In addition to the tumor cells, microvascular endothelial proliferation and smooth muscle of tumor vessel walls were frequently MT positive. The glioblastomas had a significantly higher percentage of MT positive cells compared with low grade (P < 0.0001) and anaplastic (P < 0.04) astrocytomas. MT expression in astrocytomas had no correlation with tumor recurrence. The mean Ki-67 labeling index (LI) was significantly higher in the high grade (3-4) compared with the low grade (1-2) astrocytomas. MT positive astrocytic tumors had statistically significantly higher mean Ki-67 LI compared with MT negative tumors, irrespective of histologic grade. Although the levels of MT and Ki-67 LI varied in individual tumors, the mean Ki-67 LI increased in parallel to the increasing MT staining grade, and this difference attained statistical significance only for glioblastoma. MT positive anaplastic astrocytoma and glioblastoma patients did not survive as long as the MT negative patients, although this difference attained statistical significance only for anaplastic astrocytoma. CONCLUSIONS: The current study suggests that MT might play a significant role in the growth of astrocytic tumors, with an acquired enhanced ability to produce MT as the malignant potential of a tumor increases.     

Surface-specific electrical occlusal caries diagnosis: reproducibility, correlation with histological lesion depth, and tooth type dependence

We investigated the lipid metabolism in primary cultured hepatocytes to elucidate the causes of hyperlipidemia, increased cholesteryl esters, and decreased triglyceride levels in the livers of daunomycin-nephrotic rats. The incorporation of 14C-palmitate into phospholipids and triglycerides in primary cultured hepatocytes and medium was similar in daunomycin-nephrotic and control rats. The incorporation of 14C-acetate into phospholipids, triglycerides, cholesterol, cholesteryl esters, and total fatty acids in primary cultured hepatocytes was increased in daunomycin-nephrotic rats. The radioactivity of phospholipids, triglycerides, cholesterol, cholesteryl esters, and very-low-density lipoprotein lipids in medium was increased in the hepatocytes of daunomycin-nephrotic rats using 14C-acetate as a precursor. The increased cholesterogenesis and the increased secretion of triglycerides synthesized from acetate by hepatocytes may be due to an increased cholesteryl ester content and a decreased triglyceride content in the livers of daunomycin-nephrotic rats. The increased secretion of lipids synthesized from acetate by hepatocytes may be due to increased accumulation of lipids in serum and very-low-density lipoprotein in daunomycin-nephrotic rats.     

Myoclonus associated with hyperthyroidism and thymoma: a case report

Movement disorders associated with hyperthyroidism or thymoma are extremely rare. A 52-year-old woman had developed myoclonus since June, 1989. Hyperthyroidism and thymoma were diagnosed 1 month and 9 months later. Myoclonus improved after thyroid treatment and thymectomy. If the association was not only a coincident phenomenon, autoimmunity should be considered in this previously unrecognized association.     

Multiple developmental origin of the upper incisor in mouse: histological and computer assisted 3-D-reconstruction studies

Heads of 11-15-day-old mouse embryos were cut in frontal serial sections. Early development of the maxillary incisor was analyzed using series of thick (5 and 7 microns) and semi-thin (1 micron) frontal sections and computer assisted 3-D-reconstructions of the epithelial component. The enamel organ of the mouse maxillary incisor was found to be a complex structure of multiple origin, involving several epithelial anlagen--primary dental laminae--, which could hypothetically correspond to the 5 upper incisors of early mammals. The transitory existence of at once distinct and then fusing dental primordia could reflect heterochronic changes in ontogeny which might be related to phyletic trends.     

Galectin-3 expression in human breast carcinoma: correlation with cancer histologic grade

Galectins (S-type lectins) are a family of low-molecular weight, calcium-independent, mannose-binding lectins with functions in cell growth, cell activation, cell-cell and cell-matrix adhesion including binding to carcinoembryonic antigens and laminin and metalloproteinase. Anti-galectin antisera can inhibit metastases of rat prostate cancers and human melanomas. To define the role of galectins in human breast cancer, the expression of galectin-3 were determined in 27 invasive breast cancers by immunohistochemical methods. The histologic grades of excised breast cancers were determined and immunohistochemical staining for galectin-3 (1: 1000 dilution of anti-galectin rat polyclonal antibody) was defined by scoring the intensity and distribution of staining (0-3+). The mean age of breast cancer patients was 63 years for 20 grade II breast cancers and 56 years for 7 grade III breast cancers. The mean immunohistochemical staining score for grade II breast cancers was 3. 7 (20% less than 2, 80% 3-6) and 2.5 for grade III (71.4% less than 2 and 28.6% 3-6). The galectin-3 expression pattern suggests that increasing histologic grade of breast cancer leads to reduced expression of galectin-3 and possibly reduced matrix binding and increased cancer cell motility.     

Direct measurement of brain temperature during and after intraischemic hypothermia: correlation with behavioral, physiological, and histological endpoints

The aim of the present study was to evaluate critically the protection afforded by hypothermia against ischemic injury to the hippocampus. Hypothermic treatment was applied selectively to the brain during a 5 min carotid artery occlusion in gerbils. Following a period of recovery, two independent measures were used to assess hippocampal function: (1) an open field test of spatial memory (assessment was made during the first 10 d after ischemia) and (2) measurement of evoked potentials from area CA1 in hippocampal slices (3 weeks after the ischemic episode). The functional outcome portrayed by these tests was compared to a morphological evaluation of CA1 pyramidal cells at three rostrocaudal levels. All evaluations were carried out in the same animals. We found converging evidence that intraischemic hypothermia provides virtually complete protection against a 5 min episode of cerebral ischemia. Animals treated with hypothermia performed as well as sham-operated controls in a spatial memory task, had field potentials that were indistinguishable from normal animals and CA1 cells appeared normal when assessed histologically. In contrast, ischemia at normothermia resulted in a deficit in open field behavior (p < 0.01), diminished field potentials in stratum radiatum (p < 0.01), and near total loss of pyramidal cells in dorsal CA1 (p < 0.01). There was a remarkably high correlation between these diverse measures (r ranged from 0.7 to 0.9, p < 0.01), which provides strong support for the use of hypothermia as an effective treatment for ischemia. This study introduces a novel approach for the evaluation of putative anti-ischemic treatments: combining behavioral, electrophysiological, and histological measures.(ABSTRACT TRUNCATED AT 250 WORDS)     

Differential nuclear matrix protein expression in prostate cancers: correlation with pathologic stage

PURPOSE: Nuclear Matrix Proteins (NMP) have been shown to be tissue and cell-type specific. Several unique NMPs have been investigated in various cancerous tissues, including prostate, bladder and kidney, and some are presently utilized as tumor markers. This study was aimed at characterizing the differential NMP expression in the pathologically more aggressive prostate cancers. METHODS: High resolution two-dimensional gel electrophoresis and silver staining was used to elucidate the NMP distribution of fresh prostate cancer nuclei, obtained from 39 radical prostatectomy specimens, surgically removed from men with clinically localized prostate cancer. Based on the final pathological grading, specimens were grouped according to predicted prognosis: poor--with seminal vesicle (SV) or lymph node (LN) involvement or established capsular penetration (ECP) with gleason score >7; intermediate--organ confined (OC) or focal capsular penetration (FCP) with gleason score 7 or ECP with gleason score 6; and good--with OC or FCP and gleason score <7. RESULTS: A specific charged protein (YL-1) of molecular weight 76 kD and isoelectric range 6.0-6.6 was found to be consistently present in 19 of 19 aggressive cancers. It was present only in 1 of 10 in the group with good prognosis and weakly positive in 9 of 10 in the intermediate group. CONCLUSIONS: Within this preliminary study, the expression of YL-1 appears to be related to aggressive prostate cancer, suggesting a potential marker of poor prognosis for clinically localized prostate cancer. Further characterization of the identity and function of this NMP is needed to fully ascertain its clinical potential.     

Fatty acid synthase expression in endometrial carcinoma: correlation with cell proliferation and hormone receptors

BACKGROUND: Fatty acid synthase (FAS), a biosynthetic enzyme, normally functions in the liver to convert dietary carbohydrate to fat, but it is minimally expressed in most other normal adult tissues. FAS is expressed at markedly elevated levels in subsets of human breast, ovarian, and prostate carcinomas that are associated with poor prognoses. During the menstrual cycle, the expression of FAS in the human endometrium is closely linked to the expression of the proliferation antigen Ki-67, estrogen receptor (ER), and progesterone receptor (PR). METHODS: This study reports the expression patterns of these antigens in 35 endometrial carcinomas as determined by immunohistochemical analysis. RESULTS: All cases demonstrated a close direct correlation between FAS and Ki-67 expression. Average FAS expression levels were correlated with tumor grade. Twenty-five carcinomas that were positive for ER and PR showed close correlation in expression of FAS, Ki-67, and hormone receptors. Individual tumors displayed varying degrees of heterogeneity of expression. A few well-differentiated carcinomas showed very low expression of all four antigens, similar to the antigenic profile of secretory endometrium. Nine high grade carcinomas that were negative for ER and PR also showed close correlation in expression of FAS and Ki-67 with uniformly high expression. CONCLUSIONS: These data suggest the following hypothesis: In hormone-dependent endometrial cells, FAS expression is part of the estrogen-driven cellular response that leads to proliferation; however, its linkage to proliferation is such that FAS expression is maintained in proliferating cells in endometrial carcinomas that acquire hormone independence. The use of these four antibodies as a panel may increase the diagnostic utility of ER and PR immunohistochemistry for tumor classification and prediction of the responsiveness of tumors to hormonal therapy.     

Environmental effects on cellular photosensitization: correlation of phototoxicity mechanism with transient absorption spectroscopy measurements

The presence of actin in eukaryotic nuclei and chromosomes, and especially in higher plant nuclei and chromosomes, has not been well established. We detected actin in meristematic cells of Allium cepa with indirect immunofluorescence technique and observed bright fluorescence in the intact nuclei and chromosomes, indicating that actin is present in the nuclei and chromosomes of the higher plant. We labeled sections of the meristematic cells of A. cepa with immunogold technique, gold particles were found over the whole nuclei and a number of gold particles were concentrated in condensed chromatin and nucleoli, confirming the results of the immunofluoresence observations. We treated the nuclei and chromosomes of A. cepa with DNase I and 2M NaCl and obtained DNA- and histone-depleted nuclei and chromosomes. Indirect immunofluorescence tests showed that the DNA- and histone-depleted nuclei and chromosomes reacted positively with the anti-actin antibodies. These results demonstrate that actin exists not only in intact nuclei and chromosomes, but also in DNA- and histone-depleted nuclei and chromosomes of the plant. In addition, our immuno-fluorescence tests indicate that tropomyosin is present in the nuclei and chromosomes of A. cepa.