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Neoparamoeba branchiphila n. sp., and related species of the genus Neoparamoeba Page, 1987: morphological and molecular characterization of selected strains.

A total of 18 Neoparamoeba strains were characterized both morphologically and using the SSU rRNA gene sequences as molecular markers. Nine were isolated from gills of farmed Atlantic salmon, Salmo salar L., six from sediments sampled in areas of sea-cage farms and three from net material of sea-cages. The newly obtained sequences extended substantially the dataset of Neoparamoeba strains available for phylogenetic analyses, which were used to infer taxonomic relatedness among 32 strains morphologically assigned to this genus. In addition to the N. pemaquidensis and N. aestuarina clades, phylogenetic analyses clearly distinguished a third clade with sequences from six strains. Members of this clade are characterized as representatives of a new species, N. branchiphila n. sp. The diagnostic primers for the identification of this species are introduced.

Analysis of Variance↗

Characterization of morphologic and mechanical properties of surgical mesh fabrics.

The objective of this study is to use standard testing methods to characterize the currently available synthetic mesh fabrics in terms of their chemical, physical, mechanical, and morphologic properties. Three commonly used surgical mesh fabrics, Mersilene (Ethicon), Marlex (Davol), and Teflon (USCI), were used, and the tests reveal that they differ from one another chemically as well as configurationally. The experiment included an identification of the structure of the yarn and fabrics; a measurement of the porosity, pore size and shape; and a determination of tensile and bursting strength, flexural rigidity, and wrinkle recovery. A wide variation in structure and performance was observed among the three mesh fabrics. Mersilene mesh fabrics have the highest relative porosity, while Marlex and Teflon meshes have an equivalent, but lower value. Marlex meshes have the highest tensile and bursting strength followed by Teflon and Mersilene meshes. All three meshes have one common strength characteristic--a distinctive difference in tensile strength between the wale and course directions. Marlex mesh fabrics exhibit an immense flexural rigidity and poor wrinkle recovery, due mainly to the monofilament structure of the yarn. Mersilene and Teflon mesh fabrics have similar but considerably lower, flexural rigidity than Marlex. Thus, it is evident that the chemical nature of the constituent fibers, as well as the yarn and fabric structure, have a great effect on the performance of the resulting mesh fabrics. The availability of this characterization data can serve as the basis for a surgeon's selection of the most appropriate commercial surgical mesh fabric for each case, as well as to provide a foundation for the subsequent comparison of their in vivo performance.

Materials Testing↗

Antigenic characterization of morphologically distinct Anaplasma marginale isolates using a panel of monoclonal antibodies.

The present study, describes the antigenic characterization of a Brazilian isolate of Anaplasma marginale with appendage (tail). A panel of monoclonal antibodies (McAbs) was produced and tested by the indirect fluorescent antibody test (IFAT), ELISA and Western blotting, and used to characterize two isolates of A. marginale (one with appendage and another without appendage). Among the clones produced, eight recognized antigenic proteins, with molecular weights varying from 18.4 to 66kDa. In Western blotting, the McAb reacted against a 45kDa antigen, which was shown, by the IFAT, to be located in the tail. Immunocytochemistry confirmed the tail specificity of the monoclonal reacting against the 45kDa antigen. The panel of McAb produced has a potential use in discriminating morphologically distinct A. marginale isolates. The present study, demonstrates the occurrence of antigenic diversity among Brazilian isolates of A. marginale.

Anaplasma↗

Probing the surface of polymer colloids by conductometric surfactant titration.

This work revisits the use of surfactant titrations for the characterization of latex particle surfaces. Experiments were performed to study the effect of comonomer composition and the effect of acid comonomers, and the technique is applied to the characterization of particle morphology in composite latices for several different systems. It is confirmed that the packing density of surfactant on a polymer surface is a linear function of copolymer composition. Inclusion of acid comonomers has the expected effect of decreasing the amount of surfactant adsorbed on the polymer surfaces. The usefulness of the technique in the determination of particle morphology is demonstrated, in particular toward the detection of thin layers of either seed or second-stage polymer on the particle surface which are not easily detected by other techniques such as transmission electron microscopy (TEM). Finally, it is shown that the use of acid comonomers in composite particles greatly reduces the usefulness of the surfactant titration technique for morphology characterization. A possible explanation for this effect is proposed.

Journal Article↗

Chronic pancreatitis: pathogenesis and molecular aspects.

Chronic pancreatitis (CP) is characterized by irreversible morphological and functional alterations of the pancreas presenting clinically with upper abdominal pain as well as exocrine and endocrine insufficiencies. CP is morphologically characterized by pancreatic head enlargement, calcifications of the parenchyma, cysts, and pancreatic stones. The most common etiological factor of CP in Western industrialized countries is alcohol abuse; less common factors include hereditary pancreatitis, CP due to metabolic disturbances, CP due to pancreas divisum or duodenal wall cysts, and idiopathic CP. The molecular alterations leading to the chronic inflammatory process are nor completely understood. Research during the last years, however, has elucidated that a number of growth factors and their receptors are overexpressed in CP, which is thought to contribute to the high degree of pancreatic fibrosis and to the proliferative potential of ductular cells in this disorder. In addition, gene mutations have been detected in a subgroup of CP samples underscoring the pre-malignant potential of CP. In this review we will summarize our current knowledge about pathogenic and molecular aspects of CP.

Chronic Disease↗

Magnetically responsive carboxylated magnetite-polydipyrrole/polydicarbazole nanocomposites of core-shell morphology. Preparation, characterization, and use in DNA hybridization.

Novel bis-heterocyclic mono- and dicarboxylated dipyrrole and dicarbazole monomers have been synthesized in a modular manner. Their oxidative polymerization around magnetite nanosized particles has been investigated and optimized toward new magnetic magnetite-polydipyrrole/polydicarbazole nanocomposites (NCs) of a core-shell morphology. These NCs were thoroughly characterized by FT-IR, TGA (Thermal Gravimetric Analysis), low- and high-resolution TEM/HR-TEM microscopies, and Mössbauer spectroscopy along with magnetization studies. Exploiting the versatile COOH chemistry (activation by water-soluble diimides) introduced by the polymeric shell, DNA hybridization experiments have been conducted onto NC surfaces using an efficient blue-colored HRP-based enzymatic screening biological system. Highly parallel NC-supported DNA hybridization experimentations revealed that these NCs presented an interesting potential for DNA-based diagnostic applications.

Biocompatible Materials↗

Tumor-infiltrating lymphocytes: in vitro characterization of morphological and phenotypical changes.

Mononuclear cells (tumor-infiltrating lymphocytes [TIL]) that invade tumor tissue in fresh surgical specimens were examined and characterized. Such cells were seen in virtually all 52 tumor specimens analyzed and were found to be physically attached to the tumor cells. The morphology of these cells is described with scanning and transmission electron microscopy. Phenotype analysis of these tumor-infiltrating lymphocytes was also performed. In the fresh surgical specimens, while the tumor cells are viable, TIL cells are mostly null cells (except for weak positivity for Leu-1 antigen). As the culture ages in vitro, the TIL phenotype changes drastically to a wide variety of both T and B cells as well as monocyte-macrophage lineage. While these phenotype changes occur, the tumor cells to which the TIL are attached display progressive signs of necrosis and cell death. Tumor cells without attached TIL cells are not affected by necrotic changes. It is speculated that these described changes in the phenotype and morphology of TIL may be related to acquiring cytotoxic activity against the tumor cells. An attempt to control these changes may possibly be useful for clinical application in cancer immunotherapy.

B-Lymphocytes↗

Sperm pathology: a step beyond descriptive morphology. Origin, characterization and fertility potential of abnormal sperm phenotypes in infertile men.

Sperm pathology is presented as the discipline of characterizing structural and functional deficiencies in abnormal spermatozoa. This concept complements that of sperm morphology mainly concerned with the appearance of spermatozoa. These two notions collaborate in providing correlations of prognostic value with sperm fertilizing capacity, explaining the mechanisms of sperm inefficiency, suggesting strategies to improve fertilization and opening a door to molecular genetic studies. Phenotypes of genetic origin involving sperm heads, flagella and the neck region are presented describing their clinical manifestations, sperm structure, cytochemistry and genetic background. When available, animal models are used to highlight possible genetic mechanisms. Sperm pathologies secondary to andrological conditions or environmental factors are described, stressing the non-specific nature of the sperm response to noxious agents. The available literature on the prognostic value of sperm pathologies in ICSI is also reviewed. Flagellar anomalies bear a good prognosis, but those affecting the acrosome, sperm chromatin and the neck region entail an increasing chance of failure, which highlights the differential roles played by specific sperm components in fertilization, implantation and early embryonic development. A final discussion is devoted to genetic counselling and the risks involved in using immotile or abnormal spermatozoa in assisted reproduction.

Acrosome Reaction↗

Effect of omeprazole on gastric secretion in H+,K+-ATPase and in pepsinogen-rich cell fractions from rabbit gastric mucosa.

In order to study the effects of the substituted benzimidazole omeprazole on gastric secretory functions, parietal cells and chief cells from rabbit gastric mucosa were separated and enriched by density gradient centrifugation in Percoll. H+,K+-ATPase activity, as well as a 100,000 dalton protein, was found to copurify with a cell fraction morphologically characterized as mainly parietal cells (purity approximately 65%), while pepsinogen copurified with a cell fraction morphologically characterized as chief cells (purity approximately 90%). A spontaneous pepsinogen release (9.9 micrograms/mg cell dry wt X 2 hr), unaffected by both atropine and omeprazole, was found in the chief cell fraction. The release was approximately doubled by both carbacholine (4 X 10(-5)M) and dibutyryl cAMP (db-cAMP, 10(-3)M). The cholinergic stimulation was selectively blocked by atropine, while omeprazole had no effect on pepsinogen release induced by either of the secretagogues. On the other hand, omeprazole inhibited both db-cAMP- and histamine-stimulated acid secretion quantified as [14C]aminopyrine (AP) accumulation in the parietal cell fraction. Cimetidine counteracted only acid secretion induced by histamine. These findings indicate that omeprazole has a specific effect on acid secretion, and are consonant with the hypothesis that the effect is due to H+,K+-ATPase inhibition.

Adenosine Triphosphatases↗

The use of the polymerase chain reaction and restriction fragment length polymorphism technique associated with the classical morphology for characterization of Lymnaea columella, L. viatrix, and L. diaphana (Mollusca: Lymnaeidae).

The specific identification of Lymnaeid snails is based on a comparison of morphological characters of the shell, radula, renal and reproductive organs. However, the identification is complicated by dissection process, intra and interspecific similarity and variability of morphological characters. In the present study, polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) techniques targeted to the first and second internal transcribed spacers (ITS1 and ITS2) rDNA and to the mitochondrial 16S ribosomal gene (16S rDNAmt) were used to differentiate the species Lymnaea columella, L. viatrix, and L. diaphana from some localities of Brazil, Argentina, and Uruguay as well as to verify whether the molecular results corroborates the classical morphological method.PCR-RFLP analysis of the ITS1, ITS2, and 16S using 12 restriction enzymes revealed characteristic patterns for L. columella and L. diaphana which were concordant with the classical morphology. On the other hand, for L. viatrix populations a number of 1 to 6 profiles were generated while morphology provided the species pattern results.

Animals↗

Tumor-associated desmoplasia in the rabbit mesentery characterized by morphological, biochemical and cytophotometric methods.

Intraperitoneal implantation of V2 carcinoma cells in the rabbit leads to invasion of the mesentery and to structural tissue alterations which are concomitantly of a destructive and a desmoplastic type. In this report, we describe the desmoplastic changes which are characterized by the increased formation of collagen and of proteoglycans resulting in an increased thickness of the membrane. Biochemical data indicate that the total amount of collagen increases with time after implantation, whereas the relative amount per unit of dry weight, as well as the contributions of type I (15-25%) and type III (6-8%), stay within the same range. The increased synthesis of extracellular matrix is accompanied by a change in the appearance of the fibroblasts which now show the morphologic features of synthesizing cells. Also, an appreciable number have entered the S-phase. We propose that the desmoplastic changes are tumor-associated, since implantation of epithelial cells from normal rabbit liver does not result in similar alteration. Our findings are discussed in view of the role played by tumor and/or host cells in the increased production of extracellular matrix, of possible factor(s) elaborated by the tumor cells, and of the general significance of desmoplastia for tumor spread.

Animals↗

Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation.

Apoptosis is a major form of cell death, characterized morphologically by chromatin condensation and biochemically by endonuclease cleavage of DNA into oligonucleosomal fragments. Recently, we reported that zinc arrested dexamethasone-induced apoptosis in thymocytes at an early stage, as characterized morphologically by condensation of heterochromatin into clumps abutting the nuclear membrane. In this study, we show that zinc completely inhibits endonuclease cleavage of DNA into oligonucleosomal fragments but does not prevent the cleavage of DNA into high molecular weight fragments. These results indicate that the formation of these high molecular weight fragments, which correlates with the very early morphological features of apoptosis, is a critical event in glucocorticoid-induced apoptosis. The formation of these high molecular weight fragments, despite the inhibition by zinc of the endonuclease cleavage of DNA, suggests that key enzyme(s), other than the Ca2+/Mg(2+)-dependent endonuclease, are involved at the earliest stages of induction of apoptosis.

Acetates↗

Characterization of morphologic revertants of murine and avian sarcoma virus-transformed cells.

Morphologic revertants which contain avian or murine sarcoma viruses have previously been isolated at low frequency from clonal lines of transformed mammalian cells. In the present study, these lines have been further characterized. They are indistinguishable from nontransformed parent cell lines with respect to parameters such as saturation density and colony formation in depleted medium or on monolayers of contact-inhibited cells. The rate of glucose uptake had also reverted to normal. The malignant potential of one of the revertant lines was examined and found to be markedly reduced compared to that of the corresponding transformed cells. The differences in the susceptibilities of revertant cells to retransformation by the same or other oncogenic viruses suggest that different cellular genes may be involved in expression of transformation by various tumor viruses.

Alpharetrovirus↗

Two new cell lines from B-prolymphocytic leukaemia: characterization by morphology, immunological markers, karyotype and Ig gene rearrangement.

Leukaemic cells from 2 patients with B-prolymphocytic leukaemia were immortalized in vitro by means of Epstein-Barr virus and phorbol-ester TPA. The resulting cell lines, named JVM-2 and JVM-3, have been growing continuously in liquid culture for more than one year. JVM-2 is characterized cytogenetically by t(11;14)(q13; q32), and JVM-3 by trisomy 12. The immunoglobulin (Ig) heavy- and light-chain genes showed the same pattern of rearrangement in both lines as in the original prolymphocytes from each case. The cells from these lines showed a spectrum of morphological and immunological features corresponding to different stages of B-cell maturation. The expression of Ig, IgM-lambda in JVM-2 and IgMD-K in JVM-3, changed from a predominantly membrane pattern in the original cells to a cytoplasmic one in the cell lines. By comparison with their original progenitors, the cells from both lines showed reduced reactivity with the monoclonal antibody (MAb) FMC7, and increased expression of the antigens recognized by the MAbs OKT10, alpha-Tac, FMC53 and Ki-I. The availability of cell lines from this rare type of lymphoid leukaemia offers a potential tool for the study of molecular events associated with the expression of Ig and other antigens by neoplastic cells.

B-Lymphocytes↗

Synthesis, characterization, and morphology studies of biodegradable amphiphilic poly[(R)-3-hydroxybutyrate]-alt-poly(ethylene glycol) multiblock copolymers.

Novel biodegradable amphiphilic alternating block copolymers based on poly[(R)-3-hydroxybutyrate] (PHB) as biodegradable and hydrophobic block and poly(ethylene glycol) (PEG) as hydrophilic block (PHB-alt-PEG) were successfully synthesized through coupling reaction. Their chemical structures have been characterized by using gel permeation chromatography, (1)H nuclear magnetic resonance, and Fourier transform infrared spectroscopy. Differential scanning calorimetry (DSC) analysis revealed that both PHB and PEG blocks in PHB-alt-PEG block copolymers can crystallize to form separate crystalline phase except in those with a short PEG block (M(n) 600) only PHB crystalline phase has been observed. However, due to the mutual interference from each other, the melting transition of both PHB and PEG crystalline phases shifted to lower temperature with lower crystallinity in comparison with corresponding pure PHB and PEG. The crystallization behavior of PHB block and PEG block has also been studied by X-ray diffraction, and the results were in good agreement with those deduced from DSC study. The surface morphologies of PHB-alt-PEG block copolymer thin films spin-coated on mica have been visualized by atomic force microscopy with tapping mode, indicating formation of laterally regular lamellar surface patterns. Static water contact angle measurement revealed that surface hydrophilicity of these spin-coated thin films increases with increasing PEG block content.

Chromatography, Gel↗

[Isolation, sequencing and ultrastructure of HTLV-I and HTLV-II retroviruses. Presence of the HTLV-II subtype b among Spanish intravenous drug addicts].

BACKGROUND: Six cases of HTLV-I/II infection were selected for isolation and characterization of these retrovirus. METHODS: Detection of anti-HTLV antibodies was carried out by enzyme immunoassay (EIA), immunofluorescence (IFI), and Western blot (WB). Analysis of proviral DNA was performed by PCR. Viral culture and partial sequencing of the pol and pX genes were carried out. Electron microscopy morphologically characterized the viral particles. RESULTS: Serologic study demonstrated four cases of HTLV-II, one of HTLV-I, and one non-typeable HTLV infections. This last case was confirmed as positive for HTLV-II by PCR. Five new HTLV-II and one HTLV-I infected cell lines have been established by co-culture. Electron microscopy allowed morphologic characterization of the viral particles found in the infected cells. The sequence of the five strains of HTLV-II was identical demonstrating a divergence of 0.49% in the pX region and of 4.5% in the pol region compared with the HTLV-II Mo prototype. Comparison of these sequences with those corresponding to different strains of HTLV-II isolates from American Indians (subtypes b) suggest that these Spanish strains are more closely related with the subtype b than with the subtype a (HTLV-II Mo). Genetic variability study did not reveal any change in the sequence of these stains suggesting that the variability of these retroviruses in very infrequent in the regions studied. The analysis of the pol region of the HTLV-I strain demonstrated a divergence of 3.4% with respect to the sequence of the ATK-1 prototype (Japan) and of 1.7% of the strain HS-35 (Caribbean) showing a greater relation with the Caribbean strains than with those from Japan. CONCLUSIONS: The presence of HTLV-II subtype has been confirmed among intravenous drug addicts in Spain. Isolation and characterization of the HTLV-I strain demonstrated that this also circulating around Spain despite its South American origin.

Base Sequence↗

Adult insect mushroom body neurons in primary culture: cell morphology and characterization of potassium channels.

The dissociation and maintenance in culture of cells derived from the mushroom bodies of adult crickets (Acheta domesticus) are described. This primary culture was developed in order to investigate maturation and differentiation of mushroom-body cells including Kenyon cells, the major intrinsic interneurons of mushroom bodies, which have been shown to be involved in learning and memory in insects. Three distinct cell types were observed, all identified as neural cells on the basis of their size, morphology and immunocytochemical staining with horseradish peroxidase. These cells appear to correspond to the three cell types observed in vivo: Kenyon cells, ganglion mother cells and neuroblasts. Some cells showed neurite growth, usually with long unipolar processes, occasionally with either bipolar or, more rarely, multipolar processes. Neuronal cell bodies readily formed seals with patch pipettes, allowing stable, whole-cell, patch-clamp electrophysiological recordings. Depolarization of the cell under voltage-clamp resulted in at least two types of outwardly directed potassium currents: a delayed rectifier-type of current that was sensitive to tetraethylammonium, and a cadmium-sensitive current with rapid inactivation. Neither type of current was affected by quinidine, a blocker of potassium currents recorded from pupal honeybee Kenyon cells. Other ionic currents, which have yet to be characterized, were also observed.

Animals↗

[Morphological study of Omsk hemorrhagic fever (OHF) virus with the electron microscope].

Morphological characterization of OHF virus was studied by means of thin section electron microscopy in the brains of mice infected with Omsk hemorrhagic fever virus. The crystals, microvesicles, vacuoles and as beehive structures were observed in the virus-infected cells. The possible relationship between the characteristic structures and virus morphogenesis is discussed. It is indicated that these characteristic structures may be important to morphological characterization of OHF virus.

Animals↗