bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chemically Induced Accumulation of Phospholipids in Canine Organ Systems. II. Induction by Amiodarone and Chloroquine
Authors: Maleeff Beverly E , Handler Jeffrey A , Louden Calvert S , Hart Timothy K 
Published: July 2020

Abstract
Cationic amphiphilic drugs (CAD) are structurally characterized by their hydrophobic ring structure and hydrophilic side chain. While there is little known about the effect of CAD in dogs, studies in animal models have shown that over 30 CAD with various pharmacological activities have the ability to induce phospholipid-like inclusions in peripheral blood cells and in organs such as lung, liver, eye and spleen. Two CAD used for treatment of human disease are amiodarone, prescribed for arrythmia, and chloroquine, an antimalarial. The purpose of this study was to determine if these compounds induce cytoplasmic phospholipid inclusions in peripheral blood mononuclear cells (PBMC), polymorphonuclear lymphocytes (PMN), alveloar macrophages, liver and retina in dogs.
Samples were collected from male beagle dogs after 6 weeks of oral dosing with amiodarone (75 mg/kg/day) or chloroquine (75 mg/kg/day). Buffy coats were prepared from peripheral blood and fixed with 3% glutaraldehyde in 0.1 M phosphate buffer, pH 7.2. Alveolar macrophages were recovered from bronchoalveolar lavage fluid by centrifugation and the pellets fixed with buffered 2.5% glutaraldehyde-2% formaldehyde.
Gastrin, Nitric Oxide Synthase and their Role in Ulceration and Distention of Stomach after Cisplatin and Taxol Treatment
Authors: Wang Ying , Aggarwal Surinder K 
Published: July 2020

Abstract
Cisplatin, a potent broad spectrum anti-cancer agent, has been proven effective in the treatment of different kinds of cancer, such as bladder, lung, ovarian, head, neck, etc.. Drawbacks of this chemotherapeutic drug are its toxic side-effects, which include severe nausea, vomiting, stomach distention and peptic ulcer.
Taxol is another effective chemotherapeutic agent which is usually used in conjection with cisplatin. It has demonstrated impressive activity in breast, ovarian, lung, and head and neck cancers. The toxic side-effects include nausea, vomiting, nephrotoxicity. The mechanism of these gastrointestinal side-effects are still unclear. Because of the role of NOS and gastrin on stomach smooth muscle contractility and gastroprotection, these were studied in the rats after cisplatin and taxol administration.
Wistar rats(100-150 g) were injected with cisplatin(9 mg/kg) and taxol(20 mg/kg) in five divided dosages over a period of 5 days. Rats were killed one day after the last injection. Rat stomach tissues were fixed in Bouin’s solution and processed for light microscopy.
Prohead Perestroika: Bacteriophage T7 Capsid Before and After Maturation
Authors: Cerritelli Mario E , Conway James F , Cheng Naiqian , Steven Alasdair C 
Published: July 2020

Abstract
Bacteriophage T7 provides a well defined system to study the assembly of a moderately complex virus. The mature virion consists of a 60 nm icosahedral capsid containing the linear 40 kb genome and an internal protein “core”, and a short tail. The capsid originates as a DNA-free prohead, whose assembly requires both the major capsid protein (gpl0, ∼ 420 copies) and the scaffolding protein (gp9, ∼180 copies). Native proheads incorporate an oligomeric ring of the connector/portal protein at one vertex and the inner core, but prohead-like particles assemble when gp10 and gp9 alone are expressed. In maturation -as with other dsDNA phages - the T7 prohead undergoes a major conformational change, in which the shell structure alters dramatically, and the scaffolding protein is expelled. We have used cryo-EM in combination with other techniques to study this transition and to investigate the interactions between scaffold and surface shell in the T7 prohead.
Highly purified preparations of proheads were generated from the co-expression of head and scaffolding proteins from the cloned gene in E. coli. Tail-less mature capsids as well as DNA-filled heads were prepared from infection of a T7 strain that completely lacks the tail genes.
Transport Through the Secretory Pathway of VSVG Tagged With Green Fluorescent Protein: Role of Tubulovesicular Carriers and Microtubules
Authors: Presley John , Hirschberg Koret , Cole Nelson 
Published: July 2020

Abstract
The ts045 mutant of VSV G protein has been used in numerous studies to identify biochemical and morphological properties of membrane transport, due to its reversible misfolding and retention in the ER at 40°C and ability to traffic out of the ER and into the Golgi complex upon temperature reduction to 32oC. The dynamic properties of membrane transport intermediates of the secretory pathway, including their lifetime and fate within cells, have not until now been explored due to the inability to follow transport in single living cells. Here, we attached green fluorescent protein to the cytoplasmic tail of VSV G protein in order to visualize ER-to-Golgi and Golgi-to-plasma membrane transport in living cells. VSVG-GFP expressed in Cos cells accumulated in the ER at 40°C and translocated to the Golgi complex when shifted to 32oC. Translocation of the protein was followed using time-lapse imaging of live cells on a confocal microscope. VSVG-GFP accumulated in tubulovesicular structures scattered throughout the cell upon shift from 40°C to 15°C for three hours.
Presence of P53 and Transforming Growth Factor Beta-1 in Human Cardiac Tissue With Congestive Heart Failure
Authors: Song Hong , Shuxun Ren , Fen Xia , Ruiwen Zhang , Chi-Ming Wei 
Published: July 2020

Abstract
p53 is a tumor suppressor gene which involves apoptosis and cell cycle arrest under certain stress stimulate. The expression of p53 is associated with ischemia-reperfussion induced myocardial apoptosis in rat and with the development of coronary artery restenosis after angioplasty in humans. However, the status of p53 in human cardiomyocytes and fibroblasts in congestive heart failure (CHF) remains controversial.
Transforming growth factor-beta (TGF-β) is a growth-regulating peptide that has been shown to enhance collagen production both in vivo and in vitro in cultured cells. Both p53 and TGF-β may inhibit the cell growth and induce cellular apoptosis. While the previous studies demonstrated that TGF-β1 is present in the animal myocardium, the presentation and localization of TGF-β1 in human myocardium remain unclear.
Therefore, the present study was designed to investigate the p53 and TGF-β1 in human ventricular myocardium in normal subjects and in patients with heart failure. Five normal subjects and five end-stage failing human ventricular myocardium tissue were obtained from cardiac transplantation.
Ultrastructural Observations and Heat Shock Protein 70 in Induced Unilateral Abdominal Cryptorchh) Testis in Rats
Authors: Al-Bagdadi F , Farouqi S , Stout R , Crawford P , Chandler J , Eilts B , Paccamonti D , Hosgood G , Kearny M , Duffield D , O'Donnell J 
Published: July 2020

Abstract
Infertility has been reported to be a significant consequence of cryptorchidism. Testicular cancer has been increasing since the beginning of the century (2). It has been reported in men with unilateral cryptorchidism who develop cancer that 75% of the cases are in the undescended testis. In earlier reports, investigators found that almost half of the cryptorchid testis that developed testicular cancer were abdominal (3). The extent of the degenerative changes are more extensive in the abdominal testis than the inguinal testis (1). Heat shock proteins have been detected in normal as well as in heat treated testes (4, 7). It has been stated that unchecked heat shock protein leads to irreversible damage and ultimately cell death (6). Heat shock protein is used in this study as a marker. The long range objective of this study is to determine the sequence of the ultrastructural degenerative damages of the testis over specific periods in rats as a model and relate these changes to a comparable human age.
Immunofluorescent Localization of Myosin on the Sperm Cells of Plumbago Zeylanica
Authors: Zhang Zhaojie , D. Russell Scott 
Published: July 2020

Abstract
Sperm cells in flowering plants are non-motile and are passive participants in their movement to the female reproductive cells. It is believed that actomyosin interaction may play a key role for sperm cell transmission in the pollen tube as well as in the embryo sac. However, indirect evidence has shown that the surface of sperm cells lacks amounts of myosin sufficient to support movement. Immunofluorescence microscopy was used in this study to further assess the presence of myosin on the surface of sperm cells of Plumbago Zeylanica.
Sperm cells of Plumbago Zeylanica were isolated according to published methods. Isolated sperm cells were blocked 20 min with 1% bovine serum albumin and 2% normal goat serum in phosphate buffered saline (PBS) (pH 7.3, in 15% sucrose), incubated 2 hrs in anti-myosin antibody (M-7648, Sigma Chemical Co., St. Louis, MO) diluted 1:10 with blocking solution, washed three times (5 min) with blocking solution and incubated 1 hr in 1:30 FITC-conjugated anti-rabbit IgG as secondary antibody (EY Labs, Inc., San Mateo, CA) in blocking solution. Samples were rinsed in PBS and mounted in an anti-fading solution with 1:1 PBS:glycerol with 3% n-propyl gallate.
Epidermal Keratinocyte Differentiation is Disrupted in Mice Lacking The Full Length Extracellular Calcium-Sensing Receptor
Authors: Bailey GW , Dimlich RVW , Alexander KB , McCarthy JJ , Pretlow TP , G. Kömüves László , D. Harris Jonathan , Ho Chrystal , D. Bikle Daniel 
Published: July 2020

Abstract
The importance of the extracellular calcium-sensing receptor (CaR) in the stringent control of extracellular Ca2+ concentration is well established. However, the presence of CaR in tissues not directly involved in regulating mineral ion homeostasis suggests a role for CaR in local regulation of cellular functions. Although extracellular Ca2+ regulates the differentiation of keratinocytes, the role of CaR in the epidermis is not established. In this work using knockout mice lacking full length CaR, we sought to determine the role of CaR in epidermal differentiation.
Dorsal skin of Casr−/− knockout mice lacking full length CaR, and Casr+/+ (wild type) control mice, aged 4 to 7 days after birth was fixed in 4% formaldehyde in PBS, and in 2.5% glutaraldehyde and 2% formaldehyde in 0.1 M cacodylate buffer. The samples were embedded in paraffin (for immunohistochemistry and for in situ hybridization) or in Spurr’s or LR White resins. Digoxigenin labeled antisense and sense RNA probes for loricrin and filaggrin were used for in situ hybridization.
Analysis of DNA Levels During Gonomery in Early Cleavage Divisions of the Freshwater Copepod Mesocyclops Edax
Authors: Rasch EM , Wyngaard GA 
Published: July 2020

Abstract
Gonomery, the separate grouping of maternal and paternal chromosomes during the first few mitoses following fertilization (Fig. 1) and chromatin diminution, the fragmentation and elimination of whole chromosome regions during specific stages of early cleavage in all primordial cells of the soma, occur in several species of copepods, ascarid nematodes, ciliated protozoa, Japanese hagfish 9 and a few other invertebrates. Because of the marked loss of DNA from all presumptive somatic cells by selective chromatin diminution during the 4th cleavage division in M. edax, it was of interest to determine DNA levels in chromosomes during gonomery in anaphases preceding diminution (Fig. 1). It was also important to estimate the DNA content of sperm as an index of the relative contribution by the male genome at fertilization in this species.
Mature males and egg-carrying females were fixed in 3:1 methanol/acetic acid for 3-5 mins, swollen in 45% acetic acid for 2-3 mins, squashed, frozen in liquid N2 for coverslip removal, thawed in 2 changes of absolute ETOH and air dried.
Haplotype-Specific Alphoid Oligonucleotides Can Distinguish Normal Homologous Chromosomes by Fish
Authors: Gregory Matera A , L. O'Keefe Christine 
Published: July 2020

Abstract
The field of molecular cytogenetics is presently hampered by the need for allele- and homologue-specific hybridization probes. Vast stretches of the human genome display a considerable amount of polymorphic variation between individuals. Alpha satellite DNA is a tandemly repeated sequence located at the centromeres of all primate chromosomes and is a rich source of polymorphisms. These DNA variants are phenotypically silent; they are also stable and heritable in Mendelian fashion. We have shown that we can use this genetic diversity to create homologue-specific probes using fluorescence in situ hybridization (FISH) However unlike most classical heteromorphisms, which are detected by banding techniques, our approach does not depend upon detection of gross alterations in chromosome structure, but on the basis of sequence composition. We have constructed highly-specific oligonucleotide probes that are the first reported FISH probes to discriminate between the cytogenetically indistinguishable chromosomes of normal individuals.
These sequence-specific probes have been used to follow the transmission of a single chromosome 17 through three generation families, similar to a typical polymorphic marker.
Multi-Mode Light Microscopy of Microtubule and Endoplasmic Reticulum Dynamics in Migrating Newt Epithelial Cells.
Authors: Waterman-Storer C M , Salmon E D 
Published: July 2020

Abstract
We have developed a multi-mode digital imaging system (1-3) which acquires images with a 12 bit cooled CCD camera. A multiple band pass dichromatic mirror and robotically controlled excitation filter wheels provide rapid wavelength selection for epi-fluorescence with DAPI, fluorescein or GFP and X-rhodamine fluorophores while maintaining image registration on the cooled CCD detector. Shutters select illumination either by epi-fluorescence or by transmitted light for phase contrast or DIC. A robotically controlled emission filter wheel in front of the CCD camera inserts an analyzer in the light path for DIC imaging. To maximize fluorescence light intensity, the analyzer is removed and an optical flat of equivalent optical thickness is inserted for fluorescence imaging. A slider is inserted at the field diaphragm position of the fluorescence epi-illuminator to provide in-focus slit and spot targets for 360 nm photoactivation of “caged” fluorophores. The microscope system is robotically controlled and image acquisition and analysis is performed using MetaMorph™ digital imaging software.
Three-Dimensional Analysis of Tranverse Tubules in Normal and Failing Heart: A Combined Confocal and High Voltage Electron Microscope Study
Authors: Martone M E , Edelman V M , Thor A , Young S J , Lamont S P , Ross J , Ellisman M H 
Published: July 2020

Abstract
Early electron microscopic studies documented that significant changes in the membrane systems of cardiac cells occur in both ischemic and non-ischemic heart failure. These studies relied on analysis of two-dimensional sections and although quantitative changes were observed, the overall organization of the tranverse tubules (T-tubules) and the sarcoplasmic reticulum could not be assessed. In a 3-dimensional study using high voltage electron microscopy (EM) of the T-tubules in spontaneously hypertensive rats, Nakamura and Hama (1991) observed that concomitant with an increase in surface area, the T-tubule system becomes progressively more disorganized and exhibits structural irregularities such as increased numbers of longitudinal tubules, numerous short dead end branches and complex tubular aggregates. These authors suggested that this disorganization may interfere with synchronous contraction over the entire cell.
In the present study, we examined the 3-dimensional organization of T-tubules in the left ventricle of explanted human hearts using confocal microscopy and EM tomography.
The Interactions of Fibrin and Endothelial Cells within a Fibrin Clot
Authors: Jerome W G , Hantgan RR , Handt S 
Published: July 2020

Abstract
A life threatening blood clot is the major cause of heart attacks. Thrombolytic therapy attempts to restore blood flow by activating the body's own fibrinolytic system at the site of the occlusive thrombus. However, for unknown reasons, therapy is unsuccessful in greater than 20% of patients. We have previously shown that the endothelial cells lining the wall of the vessel can play a substantial role in inhibiting clot lysis. This is due chiefly to the secretion of inhibitory molecules by endothelial cells. However, endothelial cells also have receptors for fibrin and little is known about how the direct interaction of fibrin with cells may influence lysis. To investigate this we have undertaken a series of microscopic studies to analyse the influence of endothelial cells on clot structure. We report here that endothelial cells can organize clot fibers into tight assemblies. We also show that, at least in culture, fibrin can act in concert with antithrombotic molecules to dramatically affect endothelial structure
Resources for the Study of Cellular Structure by High Voltage Electron Tomography, Serial Thin Sectioning, Specific Labeling, and Image Analysis
Authors: Mastronarde David , Kremer James , O’Toole Eileen , Morphew Mary , Ladinsky Mark , McIntosh Richard 
Published: July 2020

Abstract
We are working to improve methods for the study of cellular fine structure. Our approach is to advance each of the key steps in the preparation of specimens for EM: high quality fixation that will preserve both structure and antigenicity; methods for specific labeling; efficient acquisition of 3-D electron microscopic data; and software for 3-D reconstruction and display.
Our work on high quality structure preservation has focused on methods for fast freezing and freeze substitution. Both plunge freezing of specimens grown on coated gold grids and high pressure freezing of either cultured cells or tissue specimens have yielded well preserved material. These samples are suitable for freeze substitution fixation with either anhydrous aldehydes in acetone at -90°C, for the preservation of antigens, or aldehydes, tannic acid, OsO4, and uranyl acetate for optimal preservation the structure.
We have used a JEOL JEM-1,000 high voltage microscope to image sections about 250nm thick, employing a goniometer stage to perform dual axis tomography for 3-D reconstruction with approximately isotropic resolution at ∼7nm.
The BNL STEM Facility
Authors: Wall Joseph S , Simon Martha N , Hainfeld James F 
Published: July 2020

Abstract
The STEM facility at Brookhaven National Laboratory has been in operation since Oct. ‘77, using a custom-built instrument (STEM1) with cold field emission source, 2.5Å probe, -150°C cold stage, efficient dark field detectors and computer control & data acquisition system. A specimen changing air lock and several portable vacuum chambers permit vacuum transfer of specimens from a separate vacuum system where they were freeze dried overnight.
The large angle dark-field signal produced by the STEM is directly proportional to the total mass within the probed area. STEM mass mapping is based on this linear relationship and the fact that only specimen-specific atoms remain on the substrate after washing with volatile buffer and freeze drying. All images are digital and available via Internet. PC software can be provided for analysis.
STEM mass accuracy ranges from a fraction of a percent on well-defined individual particles such as viruses in the 50 MDa to 10 GDa range, to ∼1% around 1 MDa and ∼10% in the 50 kDa range.
The Biological Electron Microscope Facility, University of Hawai‘I at Mānoa: a Shared Microscopy Resource in the Pacific
Authors: Carvalho TM , Dunlap MF , Allen RD 
Published: July 2020

Abstract
The Biological Electron Microscope Facility (BEMF) at the University of Hawai‘i at Mānoa (UHM) is located 2400 miles over water from the next nearest research university. BEMF is a multi-user core facility, administered by the Pacific Biomedical Research Center (PBRC), an organized research unit at the UHM. The mission of the BEMF is to provide state-of-the-art instrumentation, services and training for electron microscopy to the biomedical and biological researchers in Hawai‘i and the Pacific region. The BEMF was established in 1984 under the direction of Dr. Richard D. Allen, and has since grown steadily in its instrumentation, expertise, and use. In the past 5 years it has served researchers from over 50 laboratories in PBRC and the colleges of Natural Sciences, Tropical Agriculture and Human Resources, Engineering, Medicine, and Ocean and Earth Sciences and Technology, as well as visiting investigators from other Hawai‘i, mainland and foreign institutions.
The BEMF has a full line of instrumentation for conventional transmission and field emission scanning electron microscopy as well as a complete line of instruments for cryoelectron microscopy.
Multi-Photon Microscopy, High Resolution Imaging Deep in Strongly Scattering Specimens
Authors: Denk Winfried 
Published: July 2020

Abstract
Imaging small structures substantially below the tissue surface in living specimens poses special challenges mainly because light is scattered by ever present refractive index inhomogeneities. Confocal microscoy removes the blurring caused by scattered and out-of-focus light but does so only at the expense of photodynamic damage that is often unacceptable when observing live specimens.
Multi-photon absorption microscopy[l] solves these problems because excitation is virtually limited to the focal plane. Out-of-focus photobleaching and photodamage are therefore eliminated. In scattering samples substantial improvements accrue even for the focal plane because, different from confocal microscopy, where only ballistic fluorescenc photons can be used, in the multi-photon microscope scattered photons can be utilized in addition [2-4], provided whole-field detection is used[5].
Many questions in the study of the nervous system require the investigation of intact portions of neural tissue in order to preserve the multiply branched processes of neurons, often extending over hundreds of microns, together with the local nervous circuitry.
Two-Photon Excitation Imaging of Glucose Metabolism in Living Tissue
Authors: W. Piston David 
Published: July 2020

Abstract
Two-photon excitation microscopy (TPEM) provides attractive advantages over confocal microscopy for three-dimensionally resolved fluorescence imaging and photochemistry. It provides three-dimensional resolution and eliminates background equivalent to an ideal confocal microscope without requiring a confocal spatial filter, whose absence enhances fluorescence collection efficiency. This results in inherent submicron optical sectioning by excitation alone. In practice, TPEM is made possible by the very high local instantaneous intensity provided by a combination of diffraction-limited focusing of a single laser beam in the microscope and the temporal concentration of 100 femtosecond pulses generated by a mode-locked laser. Resultant peak excitation intensities are 106 times greater than the CW intensities used in confocal microscopy, but the pulse duty cycle of 10−5 limits the average input power to less than 10 mW, only slightly greater than the power normally used in confocal microscopy. Because of the intensity-squared dependence of the two-photon absorption, the excitation is limited to the focal volume.