Cytochemical studies on nuclear DNA of four eucoccidian parasites, Isospora (Toxoplasma) gondii, Eimeria tenella, Sarcocystis cruzi and Plasmodium berghei

Parasitology ◽  
1984 ◽  
Vol 88 (1) ◽  
pp. 13-25 ◽  
Author(s):  
A. W. C. A. Cornelissen ◽  
J. P. Overdulve ◽  
M. Van Der Ploeg
Keyword(s):  
Toxoplasma Gondii ◽  
Plasmodium Berghei ◽  
Nuclear Dna ◽  
Single Cells ◽  
Fluorescence Emission ◽  
Green Light ◽  
Eimeria Tenella ◽  
Feulgen Staining ◽  
Accurate Identification ◽  
Sarcocystis Cruzi

SUMMARYFeulgen-pararosaniline(SO2) staining was performed on stages in the life-cycle of Isospora (Toxoplasma) gondii, Eimeria tenella, Sarcocystis cruzi and Plasmodium berghei. The fluorescence emission of the stained DNA in nuclei of these stages was examined and compared with absorption microscopy measurements at 560 nm (green light) of the same specimens. Accurate identification of single cells, and especially discrimination between young schizonts and young gamonts was difficult after Feulgen staining. To overcome this problem preparations were first stained with Giemsa and the cells of interest precisely located with the aid of an England finder. The same preparations were then hydrolysed and stained with Feulgen—pararosaniline(SO2), after which the previously identified cells were investigated. The DNA distribution after Feulgen staining corresponded with the shape of nuclei after Giemsa staining. DNA was present in all investigated stages of the four parasites, including macrogamonts of I. (T.) gondii and E. tenella and peripheral blood gamonts of P. berghei. Macrogamonts could be recognized, even at a stage at which they can hardly be differentiated from young schizonts in Giemsa-stained preparations, by their typical distribution pattern of Feulgen fluorescence. Feulgen fluorescence was more granular and confined to the peripheral region of the nucleus, leaving a distinct nucleolus unstained. The horseshoe-shaped nuclei typical of macrogamonts could also be observed in some second generation merozoites of E. tenella, indicating that these merozoites are already sexually differentiated. The relationship between the present cytochemical observations and the ultrastructure of the four investigated parasites is discussed.

Determination of nuclear DNA of five Eucoccidian parasites, Isospora (Toxoplasma) gondii, Sarcocystis cruzi, Eimeria tenella, E. acervulina and Plasmodium berghei, with special reference to gamontogenesis and meiosis in I. (T.) gondii

Parasitology ◽  
1984 ◽  
Vol 88 (3) ◽  
pp. 531-553 ◽  
Author(s):  
A. W. C. A. Cornelissen ◽  
J. P. Overdulve ◽  
M. Van Der Ploeg
Keyword(s):  
Mathematical Model ◽  
Toxoplasma Gondii ◽  
Plasmodium Berghei ◽  
Nuclear Dna ◽  
Eimeria Tenella ◽  
Frequency Distributions ◽  
Dna Contents ◽  
Dna Values ◽  
Sarcocystis Cruzi

SUMMARYDNA contents of individual stages of Isospora (Toxoplasma) gondii and other Eucoccida were measured after Feulgen-pararosaniline (SO2) staining either by direct microfluorometry or by scanning of microphotographic negatives. Frequency distributions were analysed using a computer program based on a mathematical model describing cell division. All stages of I. (T.) gondii, except fertilized macrogametes (2c), contained a haploid amount of DNA (1c), indicating that meiosis in I. (T.) gondii occurs during sporogony. Microgametes possessed 3·3% DNA in excess, presumably mitochondrial DNA. Nuclei of M2-and M3-merozoites differed in two characteristics: a small but distinct nucleolus was observed in almost 50% of the M2-merozoites but in none of the M3-merozoites; all M2 merozoites were strictly haploid, while all M3-merozoites were synthesizing DNA (17% above the haploid value). It may be concluded that all M2- and M3-merozoites are already sexually differentiated, i.e. are macro- and microgamontoblasts, respectively. DNA synthesis necessary for the development of the microgamont starts already in the microgamontoblast stage (M3-merozoite). M2-merozoites macrogametes, synthesize 11% extra DNA before fertilization, (after fertilization an extra amount of 12% of the diploid value was found), probably by amplification of genes for proteins which are needed for rapid maturation and later sporogony. Essentially parallel results have been found in Eimeria tenella and in crescent cystozoites of Sarcocystis cruzi. Absolute DNA values in representatives of the Eucoccida have been estimated as follows (10−15 g): I. (T.) gondii, 96; E. tenella and E. acervulina, both 75; S. cruzi, 216; Plasmodium berghei, 27. The value of the estimation of total haploid amounts as a tool in taxonomy of Eucoccida is discussed.

scholarly journals Measurement of DNA content in single cells morphologically identified on smears.

1986 ◽  
Vol 34 (10) ◽  
pp. 1253-1255 ◽  
Author(s):  
N Maruo ◽  
T Nakabo ◽  
M Kondo
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Single Cells ◽  
Green Light ◽  
Nuclear Dna Content ◽  
Absolute Methanol ◽  
Cell Populations ◽  
Bone Marrow Smear ◽  
Measuring Cell ◽  
A Cell

A method was developed for measuring the nuclear DNA content in single cells previously identified on a bone marrow smear stained by the Wright-Giemsa method. The smear was first photographed and the location of individual cells, identified by morphology, was recorded on a cell map. The smear was then bleached with 50% acid ethanol and absolute methanol, and re-stained by the Feulgen method in 0.05% pararosaniline Schiff's reagent (pH 2.3) at 7 degrees C for 10 min. Nuclear red fluorescence was observed and the intensity of this fluorescence was proportional to the amount of DNA after prior irradiation of smears with green light for 9 hr. The method is useful for measuring cell DNA content in heterogeneous cell populations when morphological cell identification is required.

scholarly journals Field Induced Slow Magnetic Relaxation in a Non Kramers Tb(III) Based Single Chain Magnet

2018 ◽  
Vol 4 (4) ◽  
pp. 59 ◽  
Author(s):  
Ajit Kumar Kharwar ◽  
Arpan Mondal ◽  
Sanjit Konar
Keyword(s):  
Density Functional ◽  
Energy Gap ◽  
Energy State ◽  
Ac Susceptibility ◽  
Fluorescence Emission ◽  
Green Light ◽  
Single Chain ◽  
Ferromagnetic Interaction ◽  
Susceptibility Data ◽  
Nitrobenzoic Acid

Herein, we report a novel Tb(III) single chain magnet with the chemical formulae [Tb(μ-OH2)(phen)(μ-OH)(nb)2]n by using 4-nitrobenzoic acid (Hnb) and 1,10-phenanthroline (phen) as ligand system. The single-crystal X-ray diffraction reveals that 4-nitrobenzoic acid acts as a monodentate ligand, water and hydroxyl ions are the bridging ligand and the phen serves as a bidentate chelating ligand. The static magnetic susceptibility measurement (from 2 K to 300 K) shows ferromagnetic interaction at very low temperature (below 6 K). The alternating current (AC) susceptibility data of the complex show temperature and frequency dependence under an applied 2000 Oe DC (direct current) field. The phen moiety behaves as an antenna and enables the complex to show the green light fluorescence emission by absorption-energy transfer-emission mechanism. To calculate the exchange interaction, broken symmetry density functional theory (BS-DFT) calculations have been performed on a model compound which also reveals weak ferromagnetic interaction. Ab initio calculations reveals the anisotropic nature (gz = 15.8, gy, gy = 0) of the metal centre and the quasi doublet nature of ground state with small energy gap and that is well separated from the next excited energy state.

scholarly journals Targeting Contrast Agents With Peak Near-Infrared-II (NIR-II) Fluorescence Emission for Non-invasive Real-Time Direct Visualization of Thrombosis

2021 ◽  
Vol 8 ◽  
Author(s):  
Kenneth S. Hettie
Keyword(s):  
Contrast Agents ◽  
Real Time ◽  
Near Infrared ◽  
Fluorescence Emission ◽  
Therapeutic Window ◽  
Direct Visualization ◽  
Spatiotemporal Resolution ◽  
Accurate Identification ◽  
Non Invasive ◽  
Nirf Imaging

Thrombosis within the vasculature arises when pathological factors compromise normal hemostasis. On doing so, arterial thrombosis (AT) and venous thrombosis (VT) can lead to life-threatening cardio-cerebrovascular complications. Unfortunately, the therapeutic window following the onset of AT and VT is insufficient for effective treatment. As such, acute AT is the leading cause of heart attacks and constitutes ∼80% of stroke incidences, while acute VT can lead to fatal therapy complications. Early lesion detection, their accurate identification, and the subsequent appropriate treatment of thrombi can reduce the risk of thrombosis as well as its sequelae. As the success rate of therapy of fresh thrombi is higher than that of old thrombi, detection of the former and accurate identification of lesions as thrombi are of paramount importance. Magnetic resonance imaging, x-ray computed tomography (CT), and ultrasound (US) are the conventional non-invasive imaging modalities used for the detection and identification of AT and VT, but these modalities have the drawback of providing only image-delayed indirect visualization of only late stages of thrombi development. To overcome such limitations, near-infrared (NIR, ca. 700–1,700 nm) fluorescence (NIRF) imaging has been implemented due to its capability of providing non-invasive real-time direct visualization of biological structures and processes. Contrast agents designed for providing real-time direct or indirect visualization of thrombi using NIRF imaging primarily provide peak NIR-I fluorescence emission (ca. 700–1,000 nm), which affords limited tissue penetration depth and suboptimal spatiotemporal resolution. To facilitate the enhancement of the visualization of thrombosis via providing detection of smaller, fresh, and/or deep-seated thrombi in real time, the development of contrast agents with peak NIR-II fluorescence emission (ca. 1000–1,700 nm) has been recently underway. Currently, however, most contrast agents that provide peak NIR-II fluorescence emissions that are purportedly capable of providing direct visualization of thrombi or their resultant occlusions actually afford only the indirect visualization of such because they only provide for the (i) measuring of the surrounding vascular blood flow and/or (ii) simple tracing of the vasculature. These contrast agents do not target thrombi or occlusions. As such, this mini review summarizes the extremely limited number of targeting contrast agents with peak NIR-II fluorescence emission developed for non-invasive real-time direct visualization of thrombosis that have been recently reported.

The heart in neuromuscular disease: primary mitochondrial diseases

ESC CardioMed ◽  
2018 ◽  
pp. 1528-1530
Author(s):  
Denis Duboc
Keyword(s):  
Neuromuscular Disease ◽  
Nuclear Dna ◽  
Single Cells ◽  
Mitochondrial Diseases ◽  
Respiratory Chain Complexes ◽  
Base Pairs ◽  
Daughter Cells ◽  
Dna Molecule ◽  
Chain Complexes ◽  
Genes Encoding

Mitochondria are responsible for energy production in most eukaryotic cells. Each cell contains at least one mitochondrion and every mitochondrion contains two to ten copies of a circular DNA molecule (mitochondrial DNA or mtDNA). Cardiomyocytes contain approximately 10,000 mtDNA copies. MtDNA is composed of around 16,500 base pairs and 37 genes encoding 13 subunits of the respiratory chain complexes I, III, IV, and V, 22 mitochondrial tRNAs and 2 rRNAs. With each cell division, mitochondria and mtDNA are randomly distributed to daughter cells. In humans, mitochondria are inherited exclusively from the mother. In healthy people mtDNA copies are usually identical at birth (homoplasmy) but with ageing, mtDNA is particularly prone to somatic mutation because, unlike nuclear DNA, it is continuously replicated, even in non-dividing tissues such as myocardium. This can lead to the propagation of somatic mutations within single cells by a process called clonal expansion. In addition, mtDNA lacks an extensive DNA repair mechanism.

scholarly journals Nano-aggregates of furan-2-carbohydrazide derivatives displaying enhanced emission with a bathochromic shift

RSC Advances ◽  
2019 ◽  
Vol 9 (62) ◽  
pp. 36097-36102
Author(s):  
Ge Ding ◽  
Xinchao Wang ◽  
Xiujuan Li ◽  
Hongpan Liu ◽  
Lunxiang Wang ◽  
...  
Keyword(s):  
Bathochromic Shift ◽  
Fluorescence Emission ◽  
Green Light ◽  
Uv Lamp

C1 exhibited obvious AIE phenomena. A change from a lack of fluorescence emission to the emission of yellow-green light under a UV lamp was observed upon the inclusion of water in the solvent.

scholarly journals Accurate identification of single-nucleotide variants in whole-genome-amplified single cells

2017 ◽  
Vol 14 (5) ◽  
pp. 491-493 ◽  
Author(s):  
Xiao Dong ◽  
Lei Zhang ◽  
Brandon Milholland ◽  
Moonsook Lee ◽  
Alexander Y Maslov ◽  
...  
Keyword(s):  
Single Cells ◽  
Whole Genome ◽  
Single Nucleotide Variants ◽  
Accurate Identification ◽  
Single Nucleotide

scholarly journals An improved method for the molecular identification of single dinoflagellate cysts

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3224 ◽  
Author(s):  
Yangchun Gao ◽  
Hongda Fang ◽  
Yanhong Dong ◽  
Haitao Li ◽  
Chuanliang Pu ◽  
...  
Keyword(s):  
Success Rate ◽  
Ultrasonic Wave ◽  
Molecular Identification ◽  
Algal Blooms ◽  
Single Cells ◽  
Pcr Amplification ◽  
Deep Understanding ◽  
Dinoflagellate Cysts ◽  
Accurate Identification ◽  
Cleaning Method

BackgroundDinoflagellate cysts (i.e., dinocysts) are biologically and ecologically important as they can help dinoflagellate species survive harsh environments, facilitate their dispersal and serve as seeds for harmful algal blooms. In addition, dinocysts derived from some species can produce more toxins than vegetative forms, largely affecting species through their food webs and even human health. Consequently, accurate identification of dinocysts represents the first crucial step in many ecological studies. As dinocysts have limited or even no available taxonomic keys, molecular methods have become the first priority for dinocyst identification. However, molecular identification of dinocysts, particularly when using single cells, poses technical challenges. The most serious is the low success rate of PCR, especially for heterotrophic species.MethodsIn this study, we aim to improve the success rate of single dinocyst identification for the chosen dinocyst species (Gonyaulax spinifera,Polykrikos kofoidii,Lingulodinium polyedrum,Pyrophacus steinii, Protoperidinium leonisandProtoperidinium oblongum) distributed in the South China Sea. We worked on two major technical issues: cleaning possible PCR inhibitors attached on the cyst surface and designing new dinoflagellate-specific PCR primers to improve the success of PCR amplification.ResultsFor the cleaning of single dinocysts separated from marine sediments, we used ultrasonic wave-based cleaning and optimized cleaning parameters. Our results showed that the optimized ultrasonic wave-based cleaning method largely improved the identification success rate and accuracy of both molecular and morphological identifications. For the molecular identification with the newly designed dinoflagellate-specific primers (18S634F-18S634R), the success ratio was as high as 86.7% for single dinocysts across multiple taxa when using the optimized ultrasonic wave-based cleaning method, and much higher than that (16.7%) based on traditional micropipette-based cleaning.DiscussionThe technically simple but robust method improved on in this study is expected to serve as a powerful tool in deep understanding of population dynamics of dinocysts and the causes and consequences of potential negative effects caused by dinocysts.

scholarly journals Cytofluorometric and cytochemical comparisons of normal and abnormal human cells from the female genital tract.

1979 ◽  
Vol 27 (1) ◽  
pp. 591-595 ◽  
Author(s):  
J E Gill ◽  
L L Wheeless ◽  
C Hanna-Madden ◽  
R J Marisa
Keyword(s):  
Acridine Orange ◽  
Female Genital Tract ◽  
Fluorescence Emission ◽  
Cell Nuclei ◽  
Feulgen Staining ◽  
Fluorescence Emission Spectra ◽  
Acridine Orange Staining ◽  
Abnormal Cell ◽  
Dnase Treatment ◽  
Abnormal Cells

Acridine orange staining of exfoliated cells from epithelial tissues facilitates discrimination between normal and abnormal cells: abnormal cells develop highly elevated nuclear fluorescence. Comparisons of acridine orange (AO) staining with propidium iodide (PI) or Feulgen staining have shown that: (a) PI staining also provides highly elevated nuclear fluorescence from abnormal cells; (b) the distributions of nuclear fluorescence following AO or PI staining were usually not significantly different as judged by the Kolmogorov-Smirnov test; (c) fluorescence emission spectra from AO and PI stained cells are consistent with the hypothesis that both fluorochromes bind to DNA within cell nuclei; (d) DNAse treatment of AO stained normal cells eliminates the nuclear fluorescence peak from slit-scan contours; RNAse treatment has no effect on nuclear fluorescence; (e) the distribution of abnormal cell nuclear fluorescence after AO staining is usually, but not always, significantly different from the distribution of abnormal cell nuclear absorbance after Feulgen staining, with relative nuclear fluorescence being greater than relative nuclear absorbance. The hypothesis currently most consistent with these results is that elevated Feulgen DNA content can account for only part of the discrimination provided by AO staining, and that the chromatin within abnormal cells is altered so as to increase accessibility of DNA to intercalating dyes.

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