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PubMed1993

Characterization of an acute micromegakaryocytic leukaemia: evidence for the pathogenesis of myelofibrosis.

The current hypothesis for the pathogenesis of myelofibrosis involves the intramedullary release of growth factors from defective or abnormal megakaryocytes. We describe a case of an acute micromegakaryocytic leukaemia, in a patient with chronic myelofibrosis, that provides additional evidence for this concept. The micromegakaryocytes, which reached 223 x 10(9)/l, were characterized morphologically by both light and electron microscopy, immunocytochemically and by platelet peroxidase activity. The cells were shown to have a mature cytoplasm, containing alpha granules and the associated proteins; vWF:Ag, fibrinogen, fibronectin and protein S. DNA analysis, by both a Seescan Solitaire Plus image analysis system and flow cytometry, revealed nuclear immaturity, with 92% of cells being diploid. Serum markers of connective tissue synthesis, namely carboxy terminal peptide of procollagen I (PICP), procollagen terminal peptide III (PIIIP) and laminin all increased significantly following transformation and were associated with an increase in platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta). These observations support the current hypothesis for bone marrow fibrosis formation and provide, for the first time, a link between in vivo growth factor release, bone marrow stromal turnover and megakaryocyte mass. In addition, the release of biologically active TGF-beta may explain both the increased fibronectin and angiogenesis characteristic of myelofibrotic bone marrow.

Female↗
PubMed1995

Alphafetoprotein-producing gastric adenocarcinoma.

We report a rare gastric tumour characterized morphologically by its hepatoid features and alpha-fetoprotein production and which presented clinically with gastric haemorrhage. Gastric fibroscopy showed a bleeding tumour of the antrum. The microscopic appearance of the tumor showed two different patterns. The most extensive presented hepatoid features. The second pattern showed undifferentiated features. The tumour cells showed immunohistochemical positivity for alphafetoprotein, EMA and p53 protein; 37% were aneuploid with a DNA index of 1.46. The serum level of alphafetoprotein was not measured before the gastrectomy but after ten days it was elevated at 1070 ng/ml. The patient died 6 months after the admission. This case provides, for the first time, information on the DNA content and the p53 expression of this unusual and aggressive variant of gastric adenocarcinoma.

Adenocarcinoma↗
PubMed1976

Short-term tissue culture of two nonsecretory human myelomas. A morphological and functional study.

Bone marrow cells from two patients without detectable monoclonal immunoglobulin (Ig) in serum and urine but with the clinical picture of plasma cell myeloma were cultivated in vitro. Immunofluorescence studies of cultured living and fixed bone marrow cells showed no signs of Ig production in one of the cases, whereas in the other case cytoplasmic kappa chains were detected, which, however, were not expressed at the surface of living cells. Cells from the later patient were also subjected to kinetic, ultrastructural, and functional studies in vitro. The fraction of myeloma cells incorporating tritiated thymidine in vitro decreased gradually during prolonged culture, indicating a continuous cell death. The morphological characterization revealed many similarities between this nonsecretory myeloma and classical myeloma, although the frequency of cells with cisternae of endoplasmic reticulum distended by a granular material was unusually high, as was the frequency of "flaming" myeloma cells. "Flaming" cells were not labeled by tritiated thymidine, suggesting that they are nonproliferative end cells. Studies of the Ig synthesis by gel diffusion analyses of supernatant and cell lysates from [14C]leucine-labeled cultures agreed with the immunofluorescence studies that the myeloma cells were nonsecretory.

Aged↗
PubMed2004

Argyrophilic grain disease: a late-onset dementia with distinctive features among tauopathies.

Argyrophilic grain disease (AgD) is a late-onset dementia morphologically characterized by the presence of abundant spindle-shaped argyrophilic grains (ArG) in neuronal processes and coiled bodies in oligodendrocytes. AgD changes consist of the microtubule-associated protein tau in an abnormally and hyperphosphorylated state and are mainly found in limbic regions, for example, in the hippocampus, the entorhinal and transentorhinal cortices and the amygdala. AgD shows a significant correlation with advancing age, and it became apparent from recent clinicopathological studies that it might account for approximately 5% of all dementia cases. Further immunohistochemical and biochemical studies revealed that AgD is a four-repeat (4R) tauopathy similar to PSP and corticobasal degeneration (CBD), but distinct from Alzheimer's disease (AD) and Pick's disease. Moreover, a common genetic background regarding the tau gene haplotype has been suggested for AgD, PSP and CBD. However, although there are currently only limited data available, AgD seems to be clinically distinct from PSP and CBD and shares rather features of (mild) AD or other forms of 'limbic' dementias, among them senile dementia with tangles and the localized form of AD.

Age of Onset↗
PubMed1996

Differential oscillatory properties of cholinergic and noncholinergic nucleus basalis neurons in guinea pig brain slice.

Evidence has suggested that the nucleus basalis magnocellularis has the potential to influence the functional state of the cerebral cortex through topographically organized, widespread projections of the cholinergic cells in that nucleus. It has also been shown that, in addition to the cholinergic neurons, other non-cholinergic magnocellular basal forebrain neurons, some of which have been identified as gamma-aminobutyric acid-ergic, project into the cerebral cortex and thus may also participate in the modulation of its activity. We have performed a comparative study of the intrinsic rhythmic properties of immunohistochemically and morphologically characterized choline acetyltransferase (ChAT)-positive and ChAT-negative cells of the nucleus basalis by means of intracellular recordings in guinea pig brain slices. Our results demonstrate that relatively large, multipolar cholinergic and non-cholinergic neurons each display differential voltage-dependent properties that allow them to discharge rhythmically in spike bursts and spike clusters, respectively, at low frequencies (< 10 Hz). Cholinergic cells display bursts of 2-4 action potentials (at approximately 200 Hz) riding on low-threshold spikes recurring at a low frequency (< 5 Hz) when depolarized from a membrane potential more negative than -55 mV and display low-frequency (< 10-15 Hz) tonic firing when depolarized from a more positive level. In contrast, non-cholinergic cells fire in a unique mode, displaying non-adapting clusters of spikes interspersed with rhythmic subthreshold membrane-potential oscillations when depolarized from levels less negative than -55 mV. The spike clusters repeat rhythmically at relatively low frequencies (2-10 Hz). The intracluster spiking frequency is relatively high and coincides approximately with that of the intervening membrane-potential oscillations (approximately 20-70 Hz). The cluster frequency of the non-cholinergic cells corresponds, in the same manner as the burst frequency of the cholinergic cells, to a delta (1-4 Hz) or theta (4-10 Hz) range of activity, whereas the intra-cluster and tonic spike frequencies of the non-cholinergic cells correspond to high beta to gamma ranges of electroencephalographic activity (19-30 Hz and 30-60 Hz, respectively). We propose that the different modes of oscillatory firing by the cholinergic and non-cholinergic basal forebrain cell populations could collectively contribute to the rhythmic modulation of slow and fast rhythms within the cerebral cortex.

Animals↗
PubMed1981

Microvessels isolated from rat brain: localization of astrocyte processes by immunohistochemical techniques.

Microvascular networks were isolated from rat telencephalon by density gradient centrifugation. The vessels prepared were characterized morphologically at the light and electron microscope level and immunohistochemically by the localization of glutamine synthetase and glial fibrillary acidic protein, two proteins found almost exclusively in astrocytes. The vast majority of the vessels prepared contained more than just endothelial cells surrounded by a basement membrane. Many arterioles were found still retaining their smooth muscle cells. Pericytes were found in association with most of the venules and many of the capillaries. Astrocyte processes remained attached to most of the microvessels. These results show that vessels prepared from rat brain still maintain most of their complex intercellular contacts and must be viewed as a heterogeneous network of cells.

Animals↗
PubMed1991

Gangliosides in human fetal brain.

The ganglioside concentration and composition were determined in 42 human fetal brains from gestational week 10 to 22, a period that is morphologically characterized by rapid neuroblast proliferation and migration. The ganglioside concentration was constant during this period, approximately 1 mumol of ganglioside sialic acid/g of fresh tissue weight. At gestational week 10 the ganglioside pattern was dominated by gangliosides of the ganglio b series, with the major ganglioside being GT1b, contributing 40% of total ganglioside sialic acid, whereas GD1b and GD3 contributed only 15 and 10%, respectively. The proportion of b series ganglioside decreased to gestational week 22, with the most pronounced relative reduction affecting GD3, but also GT1b and GD1b to a lesser extent. The ganglioside GQ1b increased in content from gestational week 10 and peaked around week 16. The proportion of GD1a increased markedly between gestational week 12 and 14 and slowly between week 14 and 18 and then increased rapidly from week 20. Ganglioside GM1 underwent a similar change. Gangliosides of the lacto series contributed 6-10% of ganglioside sialic acid between gestational week 10 and 15, and thereafter the proportion slowly decreased. 3'-isoLM1 decreased rapidly in content from gestational week 10 (20 nmol/g of fresh weight) to week 22 (less than 0.5 nmol/g of fresh weight), whereas the gangliosides of the neolacto series (3'-LM1 and 3',8'-LD1) showed a slower and less marked decline in level. The biological significance of the ganglioside changes is discussed.

Brain↗
PubMed2005

Alzheimer's disease beta-secretase BACE1 is not a neuron-specific enzyme.

The brains of Alzheimer's disease (AD) patients are morphologically characterized by neurofibrillar abnormalities and by parenchymal and cerebrovascular deposits of beta-amyloid peptides. The generation of beta-amyloid peptides by proteolytical processing of the amyloid precursor protein (APP) requires the enzymatic activity of the beta-site APP cleaving enzyme 1 (BACE1). The expression of this enzyme has been localized to the brain, in particular to neurons, indicating that neurons are the major source of beta-amyloid peptides in brain. Astrocytes, on the contrary, are known to be important for beta-amyloid clearance and degradation, for providing trophic support to neurons, and for forming a protective barrier between beta-amyloid deposits and neurons. However, under certain conditions related to chronic stress, the role of astrocytes may not be beneficial. Here we present evidence demonstrating that astrocytes are an alternative source of BACE1 and therefore may contribute to beta-amyloid plaque formation. While resting astroyctes in brain do not express BACE1 at detectable levels, cultured astrocytes display BACE1 promoter activity and express BACE1 mRNA and enzymatically active BACE1 protein. Additionally, in animal models of chronic gliosis and in brains of AD patients, there is BACE1 expression in reactive astrocytes. This would suggest that the mechanism for astrocyte activation plays a role in the development of AD and that therapeutic strategies that target astrocyte activation in brain may be beneficial for the treatment of AD. Also, there are differences in responses to chronic versus acute stress, suggesting that one consequence of chronic stress is an incremental shift to different phenotypic cellular states.

Aging↗
PubMed2005

Transgenic overexpression of prothymosin alpha induces development of polycystic kidney disease.

BACKGROUND: Polycystic kidney disease (PKD) is a genetic disorder characterized by development of renal cysts and progressive renal dysfunction. Renal tissues from both PKD patients and rodent models of PKD show elevated c-myc expression. Prothymosin alpha (ProT) is positively regulated by c-myc through binding to the E box of its promoter. Through creating transgenic mice and clinical studies, we sought to investigate whether ProT overexpression contributes to PKD development. METHODS: ProT heterozygous and homozygous transgenic mice were generated and characterized. Morphologic, histologic, immunohistochemical, and biochemical analyses of the transgenic mice were performed. RESULTS: Two transgenic lines that represented integration at two different loci of the chromosomes were generated. ProT overexpression in the kidneys of homozygous transgenic mice induced a PKD phenotype, which included polycystic kidneys, elevated blood urea nitrogen (BUN), and lethality at about 10 days of age. Similar overexpression pattern of ProT was noted in cystic kidneys of the transgenic mice as well as in human autosomal-recessive PKD (ARPKD) and autosomal-dominant PKD (ADPKD) kidneys. ProT protein levels in the kidneys and urine as well as renal mRNA level of epithelial growth factor receptor (EGFR) of homozygous ProT transgenic mice were significantly higher than heterozygous or nontransgenic littermates. Furthermore, the heterozygous transgenic mice at 17 months of age also developed mild cystic kidneys. CONCLUSION: Transgenic mice overexpressing ProT represent a novel model for PKD and may provide insights into PKD development. ProT, like c-myc and EGFR, may contribute to the development of renal cysts and may be a potential noninvasive diagnostic molecule of PKD.

Animals↗
PubMed1990

Percutaneous intracoronary ultrasound imaging: initial applications in patients.

This study was designed to evaluate the feasibility and potential clinical utility of intracoronary ultrasound imaging in patients using a 5.5 F synthetic aperture array, over-a-wire ultrasound catheter. Thirty patients underwent percutaneous intracoronary (n = 22) or peripheral (n = 8) ultrasound imaging at the time of cardiac catheterization. Ultrasound images were compared to simultaneous digital angiograms. Correlation between ultrasonic and angiographic estimation of coronary diameters was good (r = 0.80). Morphological analysis of ultrasound images obtained in patients revealed a pattern consistent with mild-to-minimal atherosclerosis, as defined by prior in vitro studies. In 15 patients, imaging was performed during intracoronary infusion of vasoactive medications (papaverine 15 mg, nitroglycerine 100 mugm, and acetylcholine 2 x 10 -5 or 2 x 10 -6M). These medications were also infused during continuous intracoronary Doppler catheter monitoring of velocity and pressure, thus allowing calculation of volume flow and coronary resistance indices. Serial determination of ultrasound-derived epicardial vessel cross-sectional area demonstrated no change (-1 +/- 1%) after papaverine, an 8 +/- 2% increase after nitroglycerine, and a 9 +/- 3% decrease after acetylcholine. Calculated resistance fell 53 +/- 4% after acetylcholine and 60 +/- 3% after papaverine. Preliminary in vitro studies using a combination angioplasty balloon/ultrasound catheter demonstrated the ability to visualize vessel dilation in real time. We conclude that intravascular ultrasound imaging will be useful for dimensional and morphological characterization of vascular disease, for beat-to-beat monitoring of vasomotion, and potentially as a real-time adjunct to therapeutic interventions.

Adult↗
PubMedDate not supplied

The journey of malaria sporozoites in the mosquito salivary gland.

The life cycle of malaria parasites in the mosquito vector is completed when the sporozoites infect the salivary gland and are ready to be injected into the vertebrate host. This paper describes the fine structure of the invasive process of mosquito salivary glands by malaria parasites. Plasmodium gallinaceum sporozoites start the invasion process by attaching to and crossing the basal lamina and then penetrating the host plasma membrane of the salivary cells. The penetration process appears to involve the formation of membrane junctions. Once inside the host cells, the sporozoites are seen within vacuoles attached by their anterior end to the vacuolar membrane. Mitochondria surround, and are closely associated with, the invading sporozoites. After the disruption of the membrane vacuole, the parasites traverse the cytoplasm, attach to, and invade the secretory cavity through the apical plasma membrane of the cells. Inside the secretory cavity, sporozoites are seen again inside vacuoles. Upon escaping from these vacuoles, sporozoites are positioned in parallel arrays forming large bundles attached by multilammelar membrane junctions. Several sporozoites are seen around and inside the secretory duct. Except for the penetration of the chitinous salivary duct, our observations have morphologically characterized the entire process of sporozoite passage through the salivary gland.

Aedes↗
PubMed1998

Bacterial isolates degrading aliphatic polycarbonates.

Bacteria that degrade an aliphatic polycarbonate, poly(hexamethylene carbonate), were isolated from river water in Ibaraki. Prefecture, Japan, after enrichment in liquid medium containing poly(hexamethylene carbonate) suspensions as carbon source, and dilution to single cells. Four of the strains, 35L, WFF52, 61A and 61B2, degraded poly(hexamethylene carbonate) on agar plate containing suspended poly(hexamethylene carbonate). Degradation of poly(hexamethylene carbonate) was confirmed by gel permeation chromatography. Besides poly(hexamethylene carbonate), the strains were found to degrade poly(tetramethylene carbonate). The strains were characterized morphologically, physiologically, and by 16S rDNA sequence analysis. Strains 35L and WFF52 were tentatively identified as Pseudomonas sp. and Variovorax sp., respectively, while strains 61A and 61B2 constitute an unidentified branch within the beta subclass of the Proteobacteria.

Bacteria↗
PubMed2004

A rare course of Whipple's disease with atypical cardiac manifestation.

Whipple's disease (WD) is a rare disorder caused by the gram-positive actinomycete Tropheryma whippelii (1). An oral route of acquisition is presumed (2). WD is a chronic systemic infection morphologically characterized by foamy PAS-positive macrophages. This case report depicts the diagnosis and clinical management of a woman who presented with atypical cardial manifestations of WD and died from severe cardiac impairment by Tropheryma whippelii which was confirmed by polymerase chain reaction. Histological assessment of myocardial specimens obtained at autopsy showed PAS-positive macrophages accumulating within foci of interstitial myocardial fibrosis. Immunohistochemical staining demonstrated myocarditis with lymphocytes and neutrophilic granulocytes, which has not been reported so far in the course of WD.

Female↗
PubMed1987

In vitro attachment of oral microorganisms to solid surfaces: evaluation of a controlled flow method.

A flow cell method was modified to provide shear-controlled experimental conditions for monitoring initial attachment and detachment of oral microorganisms to solid surfaces. Whole unstimulated human saliva was collected and circulated at a flow rate of 1 ml/min, through a cell composed of two parallel test plates. Infrared-transparent plates of medium surface energy served as test substrata in these initial calibration experiments. The plates presented a similar distribution of polar forces and dispersion forces at the surface as that of human tooth enamel and some restorative dental materials. Internal reflection infrared spectroscopy verified the presence of deposited organic material. After saliva had been circulated through the flow cells for 15 min at 37 degrees C, sterilized distilled water was introduced at the same flow rate and time of exposure to remove unattached microorganisms. Morphologic characterizations and counts of adherent Gram-stainable microorganisms were performed using incident light microscopy. Three different surface zones corresponding to the inlet area, the middle area and the outlet area of the flow cell were analyzed, and compared with enumerations of microorganisms in the whole saliva samples. Numbers of attached microorganisms in the three zones followed predictions from the laminar flow conditions, with a positive correlation shown between total numbers of microorganisms in saliva and total numbers of microorganisms attached. Cocci and rods were the only morphotypes observed on the plates, and no significant difference could be detected between the percentage of cocci and rods attached in the three different zones.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteria↗
PubMed1989

Acrosomal status quantitation in human sperm.

Acceptable methods are currently available for the rapid quantitation of acrosomal status in human sperm. Application of these methods to human sperm has led to the concept that spontaneous acrosomal loss occurs infrequently during capacitation. Induced loss occurs following sperm exposure to human follicular fluid, to zonae- or cumulus-enclosed eggs, or to calcium ionophores or other agents that modulate intracellular calcium levels. Acrosomal status quantitation may be helpful in sperm morphologic characterization or in evaluating cryodamage but has not yet been established in fertility potential measurement.

Acrosome↗
PubMed1990

Production of Schwann cell lines using a regulated oncogene.

The process of myelination in the central and peripheral nervous systems has been well characterized morphologically by a variety of techniques. It is evident from these studies that, in the peripheral nervous system myelin formation is a multistep process. Clearly, a 1:1 relationship must be established with the axon, which is followed by formation of the basal lamina and eventually myelin. Because immortalized Schwann cell lines obtained using SV40 T antigen under the control of an inducible promoter have many properties of untransfected Schwann cells in culture, including their ability to form myelin in vitro, these cells will enable us to dissect more easily the process of myelination. Having successfully immortalized rat Schwann cells without affecting their ability to differentiate fully, we are applying this approach to generate analogous cell lines from the peripheral nerves of other species such as mouse and human. Unlike rat Schwann cells, there are no known mitogens for human and mouse Schwann cells, making it impossible to expand these cell populations. The ability to produce large numbers of human Schwann cells from nerve biopsy and to analyze their biochemical properties would be of enormous value in identifying the cellular abnormalities that result in demyelinating disease. Likewise, there are several mutant mouse strains with defects in myelin formation, and cell lines from these animals would facilitate our understanding of the process leading to dysmyelination.

Animals↗
PubMed1992

Cholinergic innervation of the cerebellum of the rat by secondary vestibular afferents.

The cholinergic innervation of the cerebellar cortex of the rat was studied by immunohistochemical localization of choline acetyltransferase, radiochemical measurement of ChAT activity, and double labeling of ChAT-positive neurons with HRP injected into the cerebellum. ChAT immunohistochemistry revealed large mossy fiber rosettes as well as finely beaded terminals with different morphological characterization, laminar distribution within the cerebellar cortex, and regional differences within the cerebellum. Large "grapelike" ChAT-positive mossy fiber rosettes that were distributed primarily in the granule cell layer were concentrated, but not exclusively located, in three separate regions of the cerebellum: (1) the uvula-nodulus (lobules 9 and 10); (2) the flocculus, and (3) the anterior lobe vermis (lobules 1 and 2). Regional differences in ChAT-positive afferent terminations in the cerebellar cortex demonstrated by immunohistochemistry were confirmed by regional biochemical measurements of ChAT activity. Using ChAT immunohistochemistry in combination with HRP injections into the uvula-nodulus, we have studied the origin of the cholinergic projection. The caudal medial vestibular nucleus and to a lesser extent the nucleus prepositus hypglossus contain ChAT-positive neurons that were double labeled following HRP injections into the uvula-nodulus. We conclude that (1) there is a prominent cholinergic mossy fiber pathway to the vestibulocerebellum, (2) this pathway originates primarily in the caudal third of the medial vestibular nucleus, and (3) this cholinergic pathway likely mediates secondary vestibular information related to postural adjustment.

Afferent Pathways↗