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Cardiac troponin T in the diagnosis of myocardial injury.

In the last several decades serum levels of cardiac enzymes and isoenzymes have become the final arbiters by which myocardial damage is diagnosed or excluded. Because conventionally used enzymes are neither perfectly sensitive nor specific, there is need for a new sensitive and cardiospecific marker of myocardial damage. Cardiac troponin T (TnT) is a contractile protein unique to cardiac muscle and can be differentiated by immunologic methods from its skeletal-muscle isoform. An enzyme immunoassay specific for cardiac TnT is now available in a commercial kit for routine use. The biggest advantage of this assay is its cardiospecificity. TnT measurements, however, are also highly sensitive in diagnosis of myocardial injury and accurately discern even small amounts of myocardial necrosis. TnT measurements are, therefore, particularly useful in patients with borderline CK-MB and in clinical settings in which traditional enzymes fail to diagnose myocardial damage efficiently because of lack of specificity--for example, perioperative myocardial infarction or blunt heart trauma. TnT release kinetics reveal characteristics of both soluble, cytoplasmic, and structurally bound molecules. It starts to increase a few hours after the onset of myocardial damage and remains increased for several days. TnT allows late diagnosis of myocardial infarction. The diagnostic efficiency remains at 98% until 6 d after the onset of infarct-related symptoms. TnT is also useful in monitoring the effectiveness of thrombolytic therapy in myocardial infarction patients. The ratio of peak TnT concentration on day 1 to TnT concentration at day 4 discriminates between patients with successful (greater than 1) and failed (less than or equal to 1) reperfusion. TnT measurements are very sensitive and specific for the early and late diagnosis of myocardial damage and could, therefore, provide a new criterion in laboratory diagnosis of the occurrence of myocardial damage.

Biomarkers↗

Cytokeratin expression and hyaluronic acid production in cultures of human synovial microvascular endothelial cells: influence of cytokines and growth factors.

OBJECTIVES: To isolate and characterize human synovial endothelial cells and to determine the effects of cytokines and fibroblast growth factor on human synovial endothelial (HSE) cell hyaluronic acid production. METHODS: Endothelial cells were isolated from primary cultures of human synovial cells by fluorescent activated cell sorting based on the incorporation of a fluorescent derivative of acetylated low-density lipoprotein (DiI-Ac-LDL). Identity of endothelial cells was confirmed by positive immunostaining for von Willebrand factor (vWf), cytokeratins, endoglin, and reactivity with the lectin ulex europeans agglutinin (UEA). Hyaluronic acid production was measured by a radioligand-binding assay. RESULTS: HSE cells were isolated and maintained in long-term culture. The identity of the cultured cells as endothelial was based on uniform uptake of a (DiI-Ac-LDL), immunoreactivity for vWf, and endoglin and the binding of the lectin UEA. In addition, small blood vessels in the synovium were stained selectively with anticytokeratin antibodies K462 (cytokeratin 19 specific) and K8.13 (reactive for cytokines 1, 5, 6, 7, 8, 10, 11, and 18). Isolated HSE cells also demonstrated immunoreactivity with these cytokeratin antibodies. The cytokeratins identified by the monoclonal antibody clone K8.13 demonstrated a diffuse, fibrillar staining pattern. The cytokeratin distribution revealed with monoclonal antibody K4.62 (cytokeratin 19) was also fibrillar; however, the majority of cells also demonstrated numerous punctuate cytoplasmic vesicular structures. Treatment of HSE cells with interleukin-1 alpha (IL-1 alpha) or acidic fibroblasts growth factor (aFGF), but not tumor necrosis factor (TNF alpha), dramatically reduced the vesicular structures staining with the K4.62 antibody. HSE cells produced hyaluronic acid (HA) at a constitutive rate of 200-800 ng/10(5) cells/24 h, which could be upregulated when the cells were incubated with either IL-1 alpha or aFGF. HA production was not significantly increased when HSE cells were incubated with TNF alpha, IL-4 or interferon-gamma. CONCLUSIONS: Synovial microvascular endothelial cells produce and secrete HA and endothelial HA secretion is upregulated by IL-1 and aFGF. IL-1 and aFGF also reduce the number of vesicular-like structures immunoreactive with a monoclonal antibody to cytokeratin 19. These studies suggest that cytokine stimulation of local endothelial secretion and/or accumulation of HA may influence leukocyte adhesion to the synovial endothelium.

Adult↗

Distinguishing small cell carcinoma from non-small cell carcinoma of the lung: correlating cytologic features and performance in the College of American Pathologists Non-Gynecologic Cytology Program.

CONTEXT: The cytologic features of small cell carcinoma of the lung are well described. Nevertheless, some small cell carcinomas may be difficult to reproducibly distinguish from non-small cell carcinomas, and this distinction carries significant clinical importance. OBJECTIVE: To correlate the cytologic features of individual cases of small cell carcinoma of the lung in fine-needle aspiration specimens from the College of American Pathologists Non-Gynecologic Peer Comparison Cytology Program with the frequency of misclassification as non- small cell carcinoma. DESIGN: We reviewed 1185 interpretations of 23 different cases of small cell carcinoma in lung fine-needle aspiration specimens and correlated the cytologic features noted in these cases with performance in the program. RESULTS: Cases were divided into those that were frequently misclassified as non-small cell carcinoma (at least 10% of the responses, 11 cases) and those that were infrequently misclassified as non-small cell carcinoma (<5% of all responses, 12 cases). All cases had areas on the slides with classic features of small cell carcinoma. However, 10 of 11 cases that were frequently misclassified as non-small cell carcinoma had cells with either increased cytoplasm (4 cases), cytoplasmic globules (so-called paranuclear blue bodies) (3 cases), or apparent intracytoplasmic lumina (3 cases). These features were not identified in cases that were infrequently misclassified (P = .005). In addition, cases more frequently misclassified as non-small cell carcinoma tended to show better overall cellular and group preservation. CONCLUSIONS: Frequent misclassification of small cell carcinoma as non-small cell carcinoma in lung fine-needle aspiration specimens in this program correlates strongly with the presence of cytoplasmic features that may suggest non-small cell carcinoma or with the presence of paranuclear blue bodies. Misclassification in this program may reflect a variety of factors, including the variation in the cytologic features of individual cases, but also the lack of wide recognition that some features of non-small cell carcinoma may also be noted in well-preserved cases of small cell carcinoma.

Biopsy, Fine-Needle↗

Distinguishing primary and metastatic conventional renal cell carcinoma from other malignant neoplasms in fine-needle aspiration biopsy specimens.

CONTEXT: Differentiation of conventional renal cell carcinoma from other malignant neoplasms can be difficult, particularly with the limited sampling in fine-needle aspiration biopsies. Many studies have discussed the features of renal cell carcinoma in fine-needle aspiration biopsy specimens, but the significance of the cytologic features is not known. OBJECTIVE: To define the significant cytologic findings that aid in differentiating renal cell carcinoma from other malignant neoplasms in fine-needle aspiration biopsy specimens. DESIGN: Fine-needle aspiration biopsies from 35 patients with proven primary or metastatic conventional renal cell carcinoma and from 145 patients with proven primary or metastatic non-renal cell carcinoma malignant neoplasms were assessed for the presence or absence of the following cytologic features: the heterogeneous cell population, hemosiderin deposits, small cytoplasmic vacuoles, large cytoplasmic vacuoles, low nuclear-cytoplasmic ratio, prominent nucleoli, intranuclear inclusions, irregular nuclear membranes, and smooth nuclear membranes. Statistical analysis was performed to identify the features significant in distinguishing conventional renal cell carcinoma from non-renal cell carcinoma malignancies. RESULTS: The presence of the heterogeneous cell population, hemosiderin deposits, small cytoplasmic vacuoles, and low nuclear-cytoplasmic ratio were each highly significant in conventional renal cell carcinoma when compared with non-renal cell carcinoma malignant neoplasms (P < .001) using univariate exact analysis. Features that were identified as being predictive of conventional renal cell carcinoma using multivariable logistic regression analysis included heterogeneous cell population, small cytoplasmic vacuoles, and hemosiderin deposits (P < .05). CONCLUSIONS: The presence of the heterogeneous cell population, small cytoplasmic vacuoles, hemosiderin deposits, and a low nuclear-cytoplasmic ratio aids in differentiating conventional renal cell carcinoma from other malignant neoplasms, including morphologically similar entities such as hepatocellular carcinoma, in fine-needle aspiration biopsy specimens.

Biomarkers, Tumor↗

Transmission of non-A, non-B hepatitis agent to chimpanzees from patients of epidemic hepatitis.

Two chimpanzees were inoculated intravenously with acute-phase sera obtained from two patients with epidemic hepatitis. They developed histopathologically confirmed hepatitis. Electron microscopic examination of the liver showed peculiar cytoplasmic tubular structures in the hepatocytes. These ultrastructural findings were similar to those described for the livers of chimpanzees inoculated with the F strain of non-A, non-B hepatitis agent derived from a posttransfusion hepatitis case. The chimpanzee that had recovered from hepatitis caused by the F strain of non-A, non-B hepatitis agent was re-challenged with the serum from one of the patients. The chimpanzee developed neither clinical signs nor histological changes of hepatitis. These results suggested that non-A, non-B hepatitis agent was involved not only in post-transfusion hepatitis but also in epidemic hepatitis.

Animals↗

Multisite dephosphorylation and desensitization of conventional protein kinase C isotypes.

The generation of antisera specific for the priming phosphorylation sites on protein kinase Calpha (PKCalpha) has permitted analysis of the dephosphorylation of these sites in relation to the down-regulation of the protein. It was demonstrated that these priming sites are subject to agonist-induced dephosphorylation, consistent with inactivation of the protein. Further, the process is shown to be blocked by a PKC inhibitor, indicating a requirement for PKC catalytic activity. This was corroborated by showing that a constitutively active fragment of PKCalpha is able to stimulate the dephosphorylation of wild-type PKCalpha in transfected cells. Consistent with a membrane-traffic event, the process controlled by PKC that leads to dephosphorylation is shown to be temperature-sensitive and to correlate with transient accumulation of PKCalpha on cytoplasmic vesicular structures. It was established that the dephosphorylation of priming sites in PKCalpha is not unique and occurs with other conventional PKC isotypes, demonstrating that this is a general desensitization process for this subclass of kinases. The physiological importance of this desensitization is evidenced by the behaviour of PKCbeta1 in U937 cells, where dephosphorylation of the activation loop site is shown to be a function of cell density.

Amino Acid Sequence↗

[Role of xanthine oxidase and xanthine dehydrogenase systems in thymocyte apoptosis, induced by papaverine].

It has been established that papaverine as well as other xenobiotics (dexamethasone and nitrosodimethylamine) [figure: see text] provoked the thymocyte death like apoptosis. The increase of the quantity of double-strand, single-strand DNA breaks and low molecular weight fragments of DNA preceded cell death. In papaverine-induced process of thymocyte apoptosis the total activity of xanthine oxidase in thymocytes strongly elevated long before their death, the conversion of xanthine dehydrogenase (D-form) to xanthinoxidase (O-form) and accumulation of O-form in the cultural medium took place. Direct stimulating effect of papaverine on O-form of enzyme in thymocyte lysate was revealed. The used digitonin thymocytes were divided into cytoplasmic and structural component fractions. It was shown that about 80% of total xanthinoxidase activity was concentrated in cytoplasma while only 20% of its activity was found in structural components. More higher ratio of xanthinoxidase/xanthindehydrogenase (XO/XDH) was observed and papaverine-induced changes of these enzyme forms activities were expressed more brightly in the structural components, than in the thymocyte cytoplasma. During the process of developing thymocytes apoptosis caused by papaverine the reaction of lipid peroxidation was intensified. XO-hypoxanthin system displaying prooxidant influence on cells increased the cytotoxic effect of papaverine but the presence of allopurinol or catalase and superoxidedismutase decreased it. Besides, cytotoxic action on thymocytes of allopurinol itself as well as hypoxanthin itself was revealed.

Animals↗

The effects of chronic oral methyl mercury exposure on the lysosome system of rat kidney. Morphometric and biochemical studies.

This report describes morphometric and biochemical changes in the renal lysosome system of rats exposed to 3, 5, or 10 p.p.m. concentrations of methyl mercury hydroxide in their drinking water for 4 weeks. Increased numbers of dense, granular lysosomes, previously found to contain mercury, were observed in tubule cells of rats receiving the 3 and 5 p.p.m. dose levels but not those of the 10 p.p.m. group. Tubule cells from animals given the 10 p.p;m. dose level displayed proteinaceous vacuoles with dense crystalloid structures, apical cytoplasmic extrusion, and cellular degeneration; Mitochondrial swelling within tubule cells of treated animals showed a marked dose-response relationship. Renal microsomal activity levels of ss-glucuronidase were strongly inhibited by methyl mercury hydroxide exposure at all dose levels, whereas the activity levels of acid phosphatase were unchanged. Lysosomal beta-glucuronidase was also inhibited by methyl mercury hydroxide exposure, whereas lysosomal acid phosphatase showed approximately a 2-fold increase in activity. The results are discussed in relation to the role of lysosomes in mediating the nephrotoxic effects of methyl mercury and other toxic trace metals.

Acid Phosphatase↗

Immunohistochemical characterisation of a case of insular thyroid carcinoma.

A case of insular thyroid carcinoma arising in the right lobe of a 54-year-old male is reported. The tumour exhibited an invasive growth pattern with regional lymph node metastasis. Microscopically, the tumour was characterised by well-defined solid nests like insulae, including mature follicles containing colloid and immature follicles without colloid. Immunohistochemically, tumour cells in follicular areas predominantly exhibited immunopositivity to antithyroglobulin antibody. On the other hand, diffuse immunoreactions with anti-neuron-specific enolase (NSE), S-100 protein and Leu-7 were detected mainly in the tumour cells of solid areas. In addition, clear cytoplasmic immunoreactivity with anti-myelin basic protein (MBP) antibody was exhibited in a number of tumour cells. Ultrastructurally, many tumour cells possessed dense vacuoles, apparently containing colloid material, but intracytoplasmic neurosecretory granules were absent. The histopathological and ultrastructural characteristics of the tumour as well as its anti-thyroglobulin antibody immunoreactivity support the classical hypothesis that this neoplasm is a variant of poorly differentiated thyroid carcinoma. The positive immunohistochemical reactions for NSE, S-100, MBP and Leu-7 raise the possibility of aberrant differentiation, for example neural.

Biomarkers, Tumor↗

Aspects of the ultrastructure and function of the subcommissural organ in the Mongolian gerbil (Meriones unguiculatus).

The subcommissural organ (SCO) of the Mongolian gerbil has been studied by transmission electron microscopy. The presence of extensive rough endoplasmic reticulum replete with flocculent material, well-developed Golgi complexes, condensing vesicles, electron-opaque and dense granules in the cytoplasm of SCO cells suggests that these cells are involved in protein synthesis. The secretory granules are released at the apical surface of the ependymal cells by exocytosis into the ventricular lumen, where their product condenses to form the Reissner's fiber. From the fact that the intercellular spaces of the ependymal cells of the gerbil SCO are sealed from the ventricular lumen by tight junctions, the accumulation of secretory material within the enlarged intercellular spaces provides ultrastructural evidence of basal secretion. The secretory material observed in the prominent hypendemal intracellular canaliculi, also appears to reach the intercellular spaces. The lack of perivascular spaces and external basal lamina in the gerbil SCO apparently permits basal secretions to reach vasculature and/or leptomeningeal spaces. The presence of smooth pinocytotic and coated vesicles, multivesicular bodies and lysosomes in the cycoplasm of ependymal cells suggests that the uptake of ions and macromolecules from the cerebrospinal fluid may be an additional function of the SCO.

Animals↗

[Oogenesis of Mozambique tilapia. IV. Yolk formation].

Yolk globules in developing oocytes of Tilapia mosambique are formed by two processes: 1) biosynthetical activity of oocyte organoides; 2) vitellogenin migration by micropinocytosis and its further transformation. Undoubtedly, yolk globules of endogenic and exogenic origin are fused. The primary yolk globules are spherical, and the secondary ones are lobular. The latter originate by incorporating the former. The fast growth of the late vitellogenic stage oocytes occurs as a result of active migration of primary yolk globules into the central part of the oocyte and as their association with the secondary yolk globules. In vitellogenic oocytes of T. mosambique no yolk vesicles (cortical granules), were found by any existing methods.

Animals↗

[Endosymbiotic bacteria and their relationship with chlorella in the ciliate Climacostomum virens].

Structure of cytoplasmic bacterial symbionts of chlorella-free ciliate Climacostomum virens has been investigated. It is shown that ciliates are not able to support simultaneously growth and duplication of two different symbionts--bacteria and chlorella. Cells of C. virens lost bacterial symbionts after an artificial infection with chlorella by microinjection. Competitive relationships between two endopionts are discussed.

Animals↗

Characterization of larval haemocytes from the velvetbean caterpillar Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae).

Larval haemocytes of Anticarsia gemmatalis (Lepidoptera: Noctuidae) are presented and classified based on morphological characteristics. Haemolymph samples collected from 3rd to 6th instar larvae were observed using differential interference contrast microscopy as well as processed for transmission and scanning electron microscopy. Five general types of haemocytes were observed: prohaemocytes (Pr), plasmatocytes (Pl), granular haemocytes (GH), oenocytoids (Oe) and spherulocytes (SPh). Granular haemocytes were subdivided into two morphologically different subtypes (GH 1 and GH 2). Phenoloxidase activity was clearly observed in the Oe, and also less intensely in the Sph. The osmium/imidazole buffer technique revealed that GH 2 accumulated numerous, small lipid vesicles among their granules, while Pl contained larger but less numerous lipid inclusions. Total haemocyte counts varied between 12.3 and 20.9 x 10(3) haemocytes/microl. Pl, GH 1 and Sph were the cell types more frequently observed in all larval stages studied. GH 2 were rare in 3rd and 4th instars, becoming more numerous after the inception of the 5th instar. Populations of GH 1 and Sph maintained their proportions throughout larval development. Populations of Oe, Pl and Pr, however, presented more marked variations in their proportions. Ultrastructural studies were shown to be useful for the identification and classification of haemocytes, facilitating further analysis by light microscopy.

Animals↗

[Human epiphyseal concrements in schizophrenia].

The epiphysis is a gland containing firm extracellular bodies (brain sand) the number of which increases with age. Microscopy and roentgen microtomography showed that in some cases of schizophrenia the amount of brain sand decreases. In parallel, cytoplasm of pinealocytes appears to contain concrements of a new type--irregular hollow spheres of 0.1-1.5 microm in size. They may contain fluoride. Typical hydroxyapatite retaining organic stroma may dissolve starting from the center both in health and schizophrenia.

Adult↗

Ultrastructural pathology of neuronal membranes in the oedematous human cerebral cortex.

Surgical biopsies of frontal, parietal and temporal regions of thirty two patients with clinical diagnosis of congenital hydrocephalus, brain trauma, tumours, and vascular anomalies were examined with the transmission electron microscope. The main goal was to study the submicroscopic alterations of somatodendritic, axonal, and synaptic plasma membranes, cytomembranes, and the cytoskeleton. In both, moderate and severe oedema, fragmentation of plasma membrane, enlargement and focal necrosis of rough endoplasmic cisterns and nuclear envelope, detachment of membrane-bound ribosomes and reduction of polysome were observed. The degenerated myelinated axons exhibited discontinuities of the axolemma, disorganisation of multiple myelin lamellae, myelin sheath vacuolization, and formation of myelin ovoids. In severe oedema, synaptic disassembly was frequently found characterized by separate pre- and postsynaptic endings and loss of perisynaptic glial ensheathment. Fragmented and intact microtubules and actin-like filaments also were distinguished. The alterations of plasma membranes and cytomembranes are related with the anoxic-ischaemic conditions of brain parenchyma. The role of free radical and lipid peroxidation, disturbed energy metabolism, altered metabolic cascades, excitotoxicity, protein aggregation, and presence of extracellular oedema fluid is discussed in relation with the derangement of neuronal membranes.

Adolescent↗