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At least 1,603 records · Page 89Linked to original sources
PubMed1987

Computer automated cell size and shape analysis in cryomicroscopy.

Computer vision techniques have been developed for quantitative analysis of size and shape changes in cells frozen on a cryomicroscope. The analysis is based on implementation of standard serial edge detection algorithms in conjunction with a shape transform to isolate individual cells in complex scenes which may include adjacent and overlying ice crystals. In the present study the sensitivity of the automated analysis procedure is evaluated for images obtained by various microscope optical systems for progressive degrees of subject blurring by defocusing. Size measurements in calibration trials for freezing latex spheres with extracellular ice in the field of view were least sensitive for bright field images, although the most consistent data was obtained by differential interference contrast microscopy. In all cases phase contrast images produced the least accurate data. An example analysis is presented for the freezing of pancreas beta-cells.

Algorithms↗
PubMed2005

CPCR1, but not its interacting transcription factor AcFKH1, controls fungal arthrospore formation in Acremonium chrysogenum.

Fungal morphogenesis and secondary metabolism are frequently associated; however, the molecular determinants connecting both processes remain largely undefined. Here we demonstrate that CPCR1 (cephalosporin C regulator 1 from Acremonium chrysogenum), a member of the winged helix/regulator factor X (RFX) transcription factor family that regulates cephalosporin C biosynthesis, also controls morphological development in the beta-lactam producer A. chrysogenum. The use of a disruption strain, multicopy strains as well as several recombinant control strains revealed that CPCR1 is required for hyphal fragmentation, and thus the formation of arthrospores. In a DeltacpcR1 disruption strain that exhibits only hyphal growth, the wild-type cpcR1 gene was able to restore arthrospore formation; a phenomenon not observed for DeltacpcR1 derivatives or non-related genes. The intracellular expression of cpcR1, and control genes (pcbC, egfp) was determined by in vivo monitoring of fluorescent protein fusions. Further, the role of the forkhead transcription factor AcFKH1, which directly interacts with CPCR1, was studied by generating an Acfkh1 knockout strain. In contrast to CPCR1, AcFKH1 is not directly involved in the fragmentation of hyphae. Instead, the presence of AcFKH1 seems to be necessary for CPCR1 function in A. chrysogenum morphogenesis, as overexpression of a functional cpcR1 gene in a DeltaAcfkh1 background has no effect on arthrospore formation. Moreover, strains lacking Acfkh1 exhibit defects in cell separation, indicating an involvement of the forkhead transcription factor in mycelial growth of A. chrysogenum. Our data offer the potential to control fungal growth in biotechnical processes that require defined morphological stages for optimal production yields.

Acremonium↗
PubMed1993

Increased neurite outgrowth induced by inhibition of protein tyrosine kinase activity in PC12 pheochromocytoma cells.

Genistein and other inhibitors of protein tyrosine kinases were examined for effects on neurite elongation and growth cone morphology in the rat PC12 pheochromocytoma cell line. Genistein increased the rate of neurite elongation in PC12 cells grown on a collagen/polylysine substratum after priming with nerve growth factor (NGF), but had no effect on undifferentiated cells. Steady-state levels of phosphotyrosine-modified proteins (105, 59, 52, and 46 kDa) were reduced in NGF-primed cells by genistein treatment. The target of genistein action did not appear to be the NGF receptor/trk tyrosine kinase because the presence of NGF in cultures of NGF-primed cells was not necessary for genistein-stimulated neurite outgrowth. The tyrosine kinase inhibitors tyrphostin RG508964 and herbimycin A also increased the rate of neurite elongation in NGF-primed PC12 cells. Video-enhanced differential interference contrast microscopy revealed that growth cones of genistein-treated cells had less complex morphologies and were less dynamic than untreated cells, with short filopodia restricted to the leading edge, unlike untreated cells whose growth cones exhibited longer, more numerous filopodia and lamellipodia, which remodeled continuously. These results suggest that protein tyrosine kinase activity in PC12 cells negatively regulates neurite outgrowth and directly or indirectly affects growth cone morphology.

Adenosine Triphosphate↗
PubMed1985

Detection of the microtubule cytoskeleton of the coccidian Toxoplasma gondii and the hemoflagellate Leishmania donovani by monoclonal antibodies specific for beta-tubulin.

Seven monoclonal antibodies specific for mammalian beta-tubulin demonstrate the microtubule cytoskeleton of Toxoplasma gondii and Leishmania donovani by indirect immunofluorescence microscopy. Immunoblots of T. gondii and L. donovani proteins separated by SDS polyacrylamide gel electrophoresis confirm the specificity of the monoclonal antibodies for tubulin. Differential staining of flagellar and subpellicular microtubule populations was not seen in L. donovani with these antibodies. All seven antibodies also detected the subpellicular microtubules of T. gondii, but the polar ring and conoid of this organism was not visualized by any of them. This technique provides a rapid and specific way to assess microtubular organization in whole organisms.

Animals↗
PubMed1987

In vitro excystation of Cryptosporidium baileyi from chickens.

Release of sporozoites from the oocysts of Cryptosporidium baileyi is described from Nomarski interference-contrast microscopy. Just prior to excystation, the four sporozoites became motile and rearranged themselves within the oocyst. The sporozoites were then rapidly expelled through an opening that formed in the oocyst wall, and the residuum was either released or retained within the oocyst. Excysted sporozoites were crescent shaped and measured 5.0-9.0 microns X 1.0-1.6 micron (mean = 6.8 X 1.1 microns). Excystation occurred when sodium taurocholate or a mixture of trypsin and sodium taurocholate was present in the incubation medium. High levels of excystation occurred at 37 degrees or 40 degrees C, but excystation did not occur at 4 degrees C. The ability of biles from two avian and two mammalian hosts to produce excystation of C. baileyi was also studied. After a 2-h incubation at 40 degrees C, the percentages of excystation were 69.5% in goat bile, 45.0% in pig bile, 33.0% in chicken bile, and 34.5% in turkey bile.

Animals↗
PubMed1987

New observations on first-generation merogony of Eimeria tuskegeensis in Sigmodon hispidus.

First-generation development of Eimeria tuskegeensis was evaluated using light microscopy. Sporozoite-shaped meronts containing a prominent refractile body were observed in small intestinal cells of an experimentally infected cotton rat at 24 h post inoculation (PI). Mature spherical or subspherical meronts containing crescent-shaped merozoites were observed at 36 h PI. Refractile bodies were observed in some of these merozoites. Sporozoite-shaped meronts that were isolated from host intestinal cells and inoculated onto human fetal lung cell cultures penetrated the cultured cells by 2 h PI. A mature, subspherical, first-generation meront containing seven merozoites was observed at 9 h PI in cell culture, indicating that sporozoite-shaped meronts isolated from the host retained their infectivity.

Animals↗
PubMed1987

Effects of extracellular potassium on acid release and motility initiation in Toxoplasma gondii.

The internal pH (pHi) of Toxoplasma gondii was estimated by measuring the accumulation of the weak base 9-aminoacridine in buffers with various ionic compositions. The pHi of the metabolizing parasite increased when the extracellular K+ was elevated in alkaline medium or when the external pH (pHe) was substantially increased in medium employing high external K+ (90 mM). The parasite in mouse peritoneal fluid, or in potassium sulfate buffer (pH 8.2), where the pHi was demonstrated to be increased to 7.9, became motile when acidic buffer was substituted for the original suspension medium. This acid-induced independent movement subsided within 5 min but was repeatedly induced if the pHe was serially lowered to 6.0. Basic buffers, on the other hand, abolished motility when applied to the moving parasites. Nigericin, which is known to collapse pH gradients across the membrane, also abolished motility.

Animals↗
PubMed1988

Caryocyst-like host cell formation by Caryospora duszynskii (Apicomplexa: Eimeriidae) in human fetal lung cell cultures.

Sporozoites of the coccidium, Caryospora duszynskii, penetrated human fetal lung cell cultures but did not undergo asexual or sexual multiplication during a 29-day observation period. Beginning three days postinoculation (PI), infected host cells lost their normal elongated fibroblast-like shape and became ellipsoidal in appearance and resembled caryocysts. These caryocyst-like infected cells were observed from 3 through 29 days PI. Sporozoites remained viable throughout the study as evidenced by motility of extracellular sporozoites in infected human fetal lung cell cultures. Results of this in vitro study suggest that some species of Caryospora may form caryocysts in secondary hosts without undergoing asexual or sexual multiplication in these hosts.

Animals↗
PubMed1988

Morphology of the cyst of Giardia microti by light and electron microscopy.

Cysts of Giardia microti, isolated from feces and intestinal contents of Microtus ochrogaster, were examined by light and electron microscopy. These cysts differed morphologically from cysts of other G. duodenalis morphological types in that these cysts often contained two apparently differentiated trophozoites with mature ventral discs. Cysts more closely resembling those reported for G. lamblia and G. muris were in greater abundance in preparations made from intestinal contents and were interpreted as immature cysts. "Multiple fission" cysts, reported in G. muris and G. microti by earlier workers, were not observed; however, endosymbiotic bacteria were found in the cysts of G. microti and may have been responsible for reports of multiple fission in the cysts of Giardia.

Animals↗
PubMedDate not supplied

Morphological, small subunit rRNA, and physiological characterization of Trimyema minutum (Kahl, 1931), an anaerobic ciliate from submarine hydrothermal vents growing from 28 degrees C to 52 degrees C.

A thermophilic strain of Trimyema minutum was isolated from the hydrothermally heated sea floor at Vulcano Island (Italy) and cultivated monoxenically on Marinobacter sp. and Methanococcus thermolithotrophicus. It can be propagated strictly anaerobically and is sensitive to oxygen: if exposed to air at 48 degrees C all cells die within 60 min. It grows from 0.45-7.2% (w/v) salt and at pH 6.0-8.0. The isolate is the most extreme thermophilic ciliate which ever has been cultivated, exhibiting an optimal growth temperature of 48 degrees C (doubling time 6 h). Growth occurs between 28 degrees C and 52 degrees C. Trimyema minutum is redescribed using live observation and silver impregnation. Its morphology and the small subunit ribosomal RNA sequence is distinctly different from that of T. compressum, but morphology is highly similar to that of T. shoalsia Nerad et al. 1995, which is thus probably a junior synonym of T. minutum. To stabilize the bewildering species taxonomy in Trimyema, we suggest to recognize our population as a neotype of T. minutum.

Animals↗