bluuebunny/crossref_metadata_2025 huggingface dataset:
Neurons in Thick Hippocampal Slices: Confocal Microscopy and Three Dimensional Computer Reconstruction
Authors: Turner James N. , Szarowski Donald H. , Smith Karen , Marko Michael , Leith Ardean , Swann John W. , Inman Richard
Published: June 2020

The hippocampal formation of the brain is under intensive study as a site of seizure generation in a variety of disorders. Slices of hippocampus can be maintained in special chambers and individual neurons impaled with microelectrodes and electrically characterized. These cells exhibit seizure-like activity in response to bath applied stimuli. Pyramidal neurons have extensive dendritic trees with apical and basilar orientations, while interneurons have four first order dendrites - one projecting into each quadrant. After electrophysiologic study, individual neurons are filled with a fluorescent dye, and imaged by confocal laser scanned microscopy (CLSM). Three-dimensional (3-D) reconstruction was performed by stereo contouring and volume rendering methods. We have imaged one or more cells from 150 slices with correlative physiology on 140.

Neurons in slices (500 μm thick) were injected with lucifer yellow, fixed in 10% formalin, dehydrated in graded ethanol, and observed as whole mounts with a Bio-Rad MRC-500 and an Olympus BH-2 using an analog processor.
Behavior and Cell-to-Cell Interaction of Cultured Endothelia and Smooth Muscle Cells Seeded on Nucleopore Filter
Authors: Ghoneim Shams , Fabry Zsuzsa , Keiner Judy , Hart Michael
Published: June 2020

Vascular endothelia and smooth muscle cultures, derived from small vessels may retain some of their in vivo characteristics. It has been demonstrated by other investigators that there exists in-vivo a close relationship between endothelia cells and smooth muscle cells of most vascular beds. The myo-endothelial junction has functional implications which could explain a humoral transmition of substances between the two cell population. Rhodin demonstrated that arteriolar endothelium exhibited foot-like processes which penetrated the endothelial basement membrane and extended into smooth muscle cells of the media(1). Fawcett, believed that a pathway exists between endothelia and smooth muscle cells for the exchange of metabolites (2).

The objective of this in-vitro study was to examine the possible relationship between smooth muscle and endothelia cells in culture. Nucleopore filters with both 0.4μ and 3.0μ pores were used. They were seeded for 6 days with mouse brain endothelia cells on the top side of the filter and for 7 days with smooth muscle cells on the bottom side.
Couplings, EJSR, Gap junctions, ECC, in Ratite Heart Conduction Fibers
Authors: Sommer J.R. , Junker J. , Bossen E. , Taylor I. , Scherer B. , High T.
Published: June 2020

The ultrastructure of avian hearts has provided seminal information regarding striated muscle function. Extended junctional SR (EJSR) in avian hearts suggests that excitation-contraction coupling (ECC) is accomplished through a diffusible transmitter substance, but raises questions regarding the function of “couplings” in general. Other information gleaned from small avian hearts has elucidated adaptations of the individual and appositional geometry of the conduction fibers (CF) to functional demands imposed by different heart sizes and rates, as well as the relationship between the size of gap junctions (GJ), cell input resistance and coupling resistance between cells. The hearts of ratites (ostrich, emu, rhea) are very large (> 1 kg) and beat at about 50/min. Their working myocytes are almost identical to those found in chickens.

We investigated the following questions: 1. Do ratite CF contain “couplings”, EJSR and corbular SR? 2. Are there differences in geometry between the CF of small, fast beating, and large, slow beating hearts as is the case in mammals?
Ultrastructure Research of the Gastritis from Returning Bile
Authors: Hu Shaopeng , Wang Jianhua , Li Zhen , Chen Huei , Cu Fei , Li Nan , Mao Derhua
Published: June 2020

Gastritis from returning bile is a common disease, but the reason for the disease is not clear. As the pathologic ultrastructure research progresses, it has drawn attention to the ultrastructural change of cells in gastric mucosa by clinical workers. We observed gastric mucosa tissues of 15 patients suffering from gastritis with a transmission electron microscope (TEM) and a scanning electron microscope (SEM). It is the first report in China that fungus exists in the lamina propria of gastric mucosa tissue. The result is as follows.

The gastric mucosa tissues of 15 patients suffering from gastritis were acquired by stomachoscopy. Both TEM and SEM specimens were prepared by the usual methods. Under the TEM, the epithelial surface became higher and larger. Mitochondria of the cells were swollen and cristae were disrupted. There were vacuoles in the cells. The nucleus showed disorder, heterochromatin became darker, and nucleolae could be observed.
Calcification of Intrauterine Contraceptive Devices (IUDs)
Authors: Khan Saeed R.
Published: June 2020

Use of intrauterine contraceptive devices (IUDs) is an effective method of reproduction control. But prolonged exposure to the uterine fluid generally results in the depositon of various organic and inorganic substances on their surfaces. This crust can harbor bacteria which may be responsible for IUD associated infectious diseases. We examined used IUDs by light microscopy, scanning and transmission electron microscopy and analyzed their crust by x-ray microanalysis and x-ray diffraction to better understand the process of IUD encrustation.

Plastic IUD’s of Lippes loop type extracted after uterine exposure of various lengths of time, with obvious surface encrustations were fixed and processed for electron microscopy. A brand new Lippes loop and a used one after crust removal were exposed to vapors of 1% osmium tetroxide for 2 hours and examined by naked eye as well as analyzed by x-ray microanalysis.

IUD surfaces were unevenly encrusted, areas of heavy encrustation alternated with areas of no obvious encrustation at all.
Significance of Nonspecific Structures Present in Diagnostic Muscle Biopsies in Electron Microscopy: A Study of 168 Cases
Authors: Kalyan-Raman U.P. , Caughey R.C.
Published: June 2020

On a routine examination of 168 muscle biopsies by transmission electron microscopy, we observed various structures, like filamentous bodies in 49 cases (29%), tubular aggregates in 22 (13%), paracrystalline inclusions in 15 (9%), honeycomb arrays in 11 (6.5%), concentric laminated bodies in 10 (6%), fingerprint bodies in 6 (3.8%), and fibrous nuclear inclusions in 3 (1.8%), for a total of 116/168 or a 70% occurrence. In order to evaluate the significance and occurrence of these structures in muscle biopsies, we correlated the clinical features, light microscopic and histochemical findings on these patients.

168 muscle biopsies were studied. The muscle biopsy was obtained by using the conventional muscle clamp in all the cases. After obtaining the muscle biopsy in each case, about 1/3 of the length was frozen in liquid nitrogen at -170 degrees F to be used for histochemistry. The second 1/3 was fixed in formalin for paraffin section, and the remaining 1/3 was minced in small pieces and fixed in 2.5% glutaraldehyde for electron microscopy study.
Vascular ultrastructure of head and neck tumors
Authors: Satyanarayana U.
Published: June 2020

The Immune System protects the body against cancer either by killing the tumor cells directly or indirectly by causing damage to the tumor vasculature. Correlative Light Microscopy (LM), Scanning Electron Microscopy (SEM)and Transmission Electron Microscopy(TEM) have been done to evaluate the extent of degenerative changes in tumor vasculature.

Seven different types of premalignant and malignant lesions including leukoplakia. Squamous cell carcinoma (SCC) of Tongue, Soft palate, Tonsil, Nasopharynx, Pyriform fossa and Verrucous carcinoma of lower lip have been processed for LM, SEM and TEM by conventional procedures.

In most of the cases the following pathologic vascular changes were noted:

a) Fine gap defects (figures 1,2).

b) Coarse gap defects (figures 3,4,10-16).

c) Endothelial hypertrophy (figures 5-7).

d) widening of interendothelial spaces (figure8).

e) Sub-adventitial edema (figure 9).

Neither these pathologic changes in tumor vasculature nor the mechanism of their causation have been reported previously.
An scanning electron microscopic study of adenocarcinoma cells in pleural effusions
Authors: Shieh-Ching Yang
Published: June 2020

Cytodiagnosis of pleural effusions is very important in the clinical assessment of respiratory diseases. The existence of cancerous cells in effusions is a good evidence of indicating malignant nature of the disease process, and thus determines the treatment and prognosis of the patients. Observation of the cells derived from pleural effusion with light microscopy or transmission electron microscopy can perform detailed analysis of the cytoplasm and the nucleus. It is expected that other useful morphologic data is still contained on the plasmic membrane or cell surface. However, this can only be explored by scanning electron microscopy. In order to understand more comprehensively the surface structure of malignant cells in pleural effusion, specimens obtained from 69 patients including 26 men and 43 women were examined with scanning electron microscope in this study. The clinical diagnosis was primary adenocarcinoma of the lung in 46 patients, breast cancer in 20 patients and ovarian cancer in 3 patients. Pleural metastasis occurred in all of the patients and their chest X-ray films showed pleural effusion.
Scanning electron microscopy of human iliac bone by a simple preparation method
Authors: Takita Toshihiko , Naguro Tomonori , Kameie Toshio , Iino Akihiro , Yamamoto Kichizo
Published: June 2020

Recently with the increase in advanced age population, the osteoporosis becomes the object of public attention in the field of orthopedics. The surface topography of the bone by scanning electron microscopy (SEM) is one of the most useful means to study the bone metabolism, that is considered to make clear the mechanism of the osteoporosis. Until today many specimen preparation methods for SEM have been reported. They are roughly classified into two; the anorganic preparation and the simple preparation. The former is suitable for observing mineralization, but has the demerit that the real surface of the bone can not be observed and, moreover, the samples prepared by this method are extremely fragile especially in the case of osteoporosis. On the other hand, the latter has the merit that the real information of the bone surface can be obtained, though it is difficult to recognize the functional situation of the bone.
Defective terminal differentiation in squamous cell carcinomas and a hypothesis for oral cancer
Authors: Ahluwalia Karvita B.
Published: June 2020

Differentiation in malignant tumors is inversely related to progression, invasion and metastasis and is assessed on the basis of a marker, usually a secreted protein such as keratin. Recently, several ultrastructurally recognizable markers of differentiation were scored in breast tumors and the inadequacy in evaluating differentiation on the basis of a single marker was pointed out. The present investigations on squamous cell carcinomas of the tongue, buccal mucosa and nasopharynx (total of 15 samles), erythroplakia (one) and leukoplakia (3) have revealed a structural pathway underlying defective terminal differentiation in malignant cells.

There are two types of protein synthesis in cells. One, on free cytoplasmic polysomes where enzymes for metabolic reactions and cytoskeletal proteins are thought to be synthesised. The second concerns biosynthesis of secretory proteins, membranes and lysosomal enzymes and takes place in the endomembrane system (EMS). The EMS is constituted by the nuclear envelope, rough endoplasmic reticulum (RER), Golgi apparatus, transport vesicles and lysosomes.
Ultrastructure of a Paratesticular Rhabdomyosarcoma
Authors: Siew Shirley , Katlein Stanley M.
Published: June 2020

Rhabdomyosarcoma, a malignant tumor of striated muscle, was described first in 1849 by Rokitansky. Since that time, another 150 cases have been documented. In recent years, it has been recognized that the paratesticular location of rhabdomyosarcoma is an important single site, particularly in adolescents. As a combination of surgery and chemotherapy can affect a cure, a definitive diagnosis should be established as early as possible.

We report the case of an 18 year old boy, who presented with a scrotal mass, which was removed surgically, with the clinical impression of a hydrocele. On gross examination, the mass measured 4cm. He showed the presence of cystic change,foci of hemorrhage and solid white neoplastic tissue. Sheets of large polygonal cells were noted on histologic examination and there was infiltration of the epididymis.

Microscopic examination of Epon embedded material cut at micron and stained with to luidine blue confirmed the presence of a diffuse infiltrate of large polygonal cells. The cells were arranged in sheets, which were separated by irregular fibrous bands.
Ultrastructural effects of lipid peroxidation on the cardiac sarcolemma of the isolated rat heart
Authors: Oguro T. , Onodera T. , Ashraf M.
Published: June 2020

Previously we reported that hydrogen peroxide (H2O2) is primarily cytotoxic to the cardiac cell membranes presumably through lipid peroxidation (LP). However, the effect of LP on the altered cell membrane permeability remains unknown. In this study, we evaluated the ultrastructural effects of LP on the sarcolemma of the isolated rat heart using H2O2and N,N’-diphenyl-p-phenylene diamine (DPPD), an antilipid peroxidative agent.

The hearts were perfused with 300 μM H2O2for 15 minutes. In the other experiment the hearts were perfused with 2.5 μM DPPD for 20 minutes prior to perfusion with H2O2for 15 minutes. Coronary effluent was collected at the end of the equilibration period and during H2O2treatment for malondialdehyde (MDA) determination. Two minutes prior to the termination of the experiment, horseradish peroxidase (HRP: 220 U/mg × 55 mg) was perfused to assess the enhanced permeability of the cell membranes. The tissue slices were incubated in Graham-Karnovsky medium for one hour followed by postfixation with buffered 1% OsO4.
Ineffective Thrombopoiesis
Authors: Radley JM , Ellis SL
Published: June 2020

In effective thrombopoies is has been inferred to occur in several disease sates from considerations of megakaryocyte mass and platelet kinetics. Microscopic examination has demonstrated increased numbers of megakaryocytes, with a typical forms particularly pronounced, in primary myelofibrosis. It has not been documented if megakaryocyte ever fail to reach maturity in non-pathological situations. A major difficulty of establishing this is that the number of megakaryocytes normally present in the marrow is extremely low. A large transient increase in megakaryocytopoiesis can how ever be induced in mice by an injection of 5-fluorouracil. We have utilised this treatment and report here evidence for in effective thrombopoies is in healthy mice.

Adult mice were perfused (2% glutaraldehyde in 0.08M phosphate buffer, pH 7.4) 8 days following an injection of 5-fluorouracil (150mg/kg). Femurs were subsequently decalcified in 10% neutral E.D.T.A. and embedded in Spurrs resin. Transverse sections of marrow revealed many megakaryocytes at various stages of maturity. Occasional megakaryocytes (less than 1%) were found to be under going degeneration prior to achieving full maturation and releasing cytoplasm as platelets. These cells were characterized by a peripheral rim of dense cytoplasm which enveloped a mass of organelles and vacuoles (Fig. 1). Numerous microtubules were foundaround and with in the organelle-rich zone (Fig 2).
Ultrastructural study of the effect of POLYCTFE on primate liver
Authors: Mattie D.R. , Maslanka J.J. , Chase M.R.
Published: June 2020

Polychlorotrifluoroethylene (POLYCTFE) existing as oligomers of 3 and 4 units, is a candidate base stock for a nonflammable hydraulic fluid for advanced aircraft. POLYCTFE caused significant changes in the livers of rats after repeated inhalation exposure. Ultrastructural studies of the livers from rats exposed to vapors of POLYCTFE oligomers for 90-days revealed an increase in the number of peroxisomes, dose-dependent mitochondrial swelling and a dose-dependent increase in smooth endoplasmic reticulum. The objective of this study was to ultrastructurally examine hepatocytes from the livers of primates exposed to POLYCTFE and compare the exposed primate hepatocytes to those from rat livers, in order to determine the relevance of the effect of POLYCTFE on rat livers to human livers.

Male rhesus monkeys weighing 8 to 10 kg were dosed by gavage daily for 15 days with 0.725.g/kg POLYCTFE. Four animals received POLYCTFE and 3 received a comparable amount of water.
Relationship between electron microscopic changes in alveolar septa and extravascular lung water volume
Authors: Oda Hiroshi , Arakawa Michio , Kambara Kenjiro , Nakahara Koji , Segawa Takashi , Ando Fumio , Kawada Tomoo , Hirakawa Senri , Emura Shoichi , Shoumura Shizuko , Isono Hideo
Published: June 2020

Lung water accumulates partly in the alveolar septa. However, morphological changes in alveolar septa have never been analyzed quantitatively nor correlated with lung water volume changes. We aimed to delineate these unsolved questions.

Twenty-four mongrel dogs were studied under general anesthesia and spontaneous breathing. Extravascular lung thermal volume was measured with double indicator dilution technique using heat and Indocyanine green. Experimental groups were as follows: Group C; 6 control dogs, Group D-30; 100 ml/kg of dextran-70 over 30 min in 6 dogs, Group D-90; 100 ml/kg of dextran-70 over 30 min and waiting for another 60 min in 6 dogs, Group A; 100 mg/kg of alloxan. After final measurement of extravascular lung thermal volume, we immediately killed the dogs with the lungs inflated and we removed the lungs for lung water measurement with direct destructive method. Samples for electron microscopic examination were obtained by excising tissues from the surface of right lower lobe where edema appeared severe. Twenty electron micrographs were prepared from each dog without prior knowledge of the experimental intervention.
Autotransplantation of adrenal cortex: An autoradiographic study
Authors: Magalhães Maria C. , Vendeira Pedro , Magalhães Manuel
Published: June 2020

In previous experiments we demonstrated that 4, 12 and 24 weeks after rat adrenal cortex autotransplantation, the animals survived and the autotransplant was easily reproducible. It was observed that 4 weeks after transplantation the plasma corticosterone concentration (0.20 μg/ml) was already comparable to that in controls and that the light and electron microscopy features of the cells of the glomerulosa and fasciculata zones were similar to those of adrenal cells in control animals.

In order to clarify the origin of glandular cells in the autotransplanted animals, autoradiographic experiments were made. Four male rats with body weights ranging from 180to195 gm underwent bilateral adrenalectomy via two dorsal incisions made along the lateral border of the dorsal muscle mass. Pieces of adrenals were autotransplanted immediately under the skin of the dorsal region. Four animals were “sham-operated” without removal of adrenals and served as controls. The animals were fed “ad libitum” and with saline solution, and were sacrificed at 2,3,7 and 15 days. 3H-Thymidine (1 μCi/gm body weight) was administrated intraperitoneally to each animal two hours before the sacrifice. Necropsy was carried out in all animals in order to search for accessory adrenals. The autotransplanted pieces were removed, fixed and processed for LM and EM morphologic and autoradiographic studies. For LM autoradiographic studies Ilford K5 nuclear emulsion was used, and the sections exposed for 33 days; for EM autoradiographic studies Ilford L4 nuclear emulsion was used, the sections exposed for 90 days, and the radio autographs developed with Kodak D 19b.
Ultrastructural evidence of Human Immunodeficiency Virus (HIV-1) multiplication in cultured endothelial cells (EC) of the human hepatic sinusoid
Authors: Gendrault Jean-Louis , Steffan Anne-Marie , Schweitzer Christine , Schmitt Marie-Paule , Royer Cathy , Jaeck Daniel , Kirn André , Aubertin Anne-Marie
Published: June 2020

Although hepatic sinusoidal alterations have often been observed in AIDS patients, the infection of the EC of the liver with HIV has never been accounted for. The present study aims at demonstrating that EC isolated from the hepatic sinusoid may be permissive for HIV-1.

EC were isolated from surgical biopsies obtained from HIV-seronegative patients suffering from liver cancer. The cells were prepared by collagenase dissociation of the tissue and centrifugal elutriation. The cells were cultured in 24-well culture plates in the medium previously described for 5-6 days before being infected with 2.104 syncytia-forming units of HIV-1 (isolate HTLV-III B) per well. They were fixed after 7, 14 and 19 days of infection and routinely prepared for TEM and SEM. The presence in the EC of the von Willebrand factor was tested by immunofluorescence (FVIII.rAg, Behring).

Typical syncytia resulting from the infection were observed (Figs. 1, 2), their endothelial origin being demonstrated by the presence of the von Willebrand factor (Fig. 1). This origin could be further assessed by observing characteristic fenestrae (Figs. 3, 4) and Weibel-Palade bodies (Fig. 5).
Levels of expression of Human Immunodeficiency Virus antigens in lymphocytes from healthy carriers by monoclonal antibody probing and backscattered electron imaging
Authors: Herrera M.I. , Cuevas L. , Santa-María I. , Nájera R.
Published: June 2020

The knowledge about the state of viral replication over the course of infection with the Human Immunodeficiency Virus type 1 (HIV-1) is still incomplete in spite of its important implications for the pathogenesis and transmission of AIDS. Silent HIV-1 infections might take place after the initial encounter with the virus; infected persons may remain seronegative for prolonged periods, suggesting that in this state very little, if any, viral RNA or protein is made and infectious virus is not released. In anti-HIV seropositive yet healthy individuals (“healthy carriers”), earlier studies using viral isolation techniques from peripheral blood mononuclear cells (PBMC) have also contributed to the notion that the number of infected cells in this subjects is low (at most 1 in 100 in 1 case only). Present reports on the presence of free virus in plasma or on antigen expression in sera or in cells are still scarce and controversial.
Immunolocalization of human alveolar macrophage prostaglandin H synthase using lnm gold probe and silver enhancement techniques
Authors: Yi Hong , Pueringer Robert J. , Moore Kenneth C. , Hunninghake Gary W.
Published: June 2020

Human alveolar macrophages (HAMs) stimulated with lipopolysaccharide (LPS) produce large amounts of prostaglandin E2(PGE2), a potent anti-inflammatory mediator, for prolonged periods of time. We have previously demonstrated that this prolonged PGE2production is delayed until 6 hours after LPS stimulation and is in part dependent on new synthesis of the first committed enzyme, prostaglandin H (PGH) synthase. To determine if the new synthesis of PGH synthase is compartmentalized to a specific cellular structure, we immunolocalized PGH synthase with light and electron microscopy at 24 hours after stimulating HAMs.

In order to ensure the antigenicity of the enzyme for antibody recognition and better penetration of the antibody conjugates, pre-embedding labelling with 1 nm gold probe and silver enhancement techniques were employed. HAMs were isolated from normal volunteers by bronchoalveolar lavage and were stimulated with LPS (10 ug/ml) for 24 hours. PGE2was measured by radioimmunoassay (RIA). Both unstimulated and stimulated cells were fixed with PLP (paraformaldehyde-lysine-periodate) fixative and gently permeabilized with 0.01% Triton X-100. We used 1 nm gold conjugated goat antirabbit IgG (Janssen) at a final dilution of 1:100 to bind to the antibodies specific for PGH synthase from rabbit antisera. The cells were then post-fixed and silver-enhanced using the Inten SEM kit (Janssen) for light microscopy and Danscher’s method for electron microcopy.
Motility of leukemic lymphocytes
Authors: Raje Manoj , Ahluwalia Karvita B.
Published: June 2020

In Acute Lymphocytic Leukemia motility of lymphocytes is associated with dissemination of malignancy and establishment of metastatic foci. Normal and leukemic lymphocytes in circulation reach solid tissues where due to in adequate perfusion some cells get trapped among tissue spaces. Although normal lymphocytes reenter into circulation leukemic lymphocytes are thought to remain entrapped owing to reduced mobility and form secondary metastasis. Cell surface, transmembrane interactions, cytoskeleton and level of cell differentiation are implicated in lymphocyte mobility. An attempt has been made to correlate ultrastructural information with quantitative data obtained by Laser Doppler Velocimetry (LDV). TEM of normal & leukemic lymphocytes revealed heterogeneity in cell populations ranging from well differentiated (Fig. 1) to poorly differentiated cells (Fig. 2). Unlike other cells, surface extensions in differentiated lymphocytes appear to originate by extrusion of large vesicles in to extra cellular space (Fig. 3). This results in persistent unevenness on lymphocyte surface which occurs due to a phenomenon different from that producing surface extensions in other cells.
LDL receptor and /ßVLDL binding: Co-localization on whole-mount pigeon monocyte derived macrophages
Authors: Jones Nancy L. , Lewis Jon C.
Published: June 2020

Monocyte derived macrophage foam cells, hallmark to the disease atherosclerosis, can be formed in vitro by cell exposure either toβVLDL, a naturally occurring lipoprotein, or experimentally modified lipoproteins such as acetylated LDL. Binding kinetics of pigeon βVLDL suggest at least two binding sites on pigeon peritoneal macrophages, and these two kinetically identified sites are consistent with two ultrastructurally distinct uptake mechanisms. One of these sites, the * pigeon βVLDL receptor”, is believed to be a form of the native LDL receptor. This βVLDL/LDL receptor relationship on monocyte derived macrophages is evidenced by 40% overlap in receptor competition between LDL and βVLDL.We have previously reported two ligand induced regions for βVLDL binding sites on pigeon monocyte derived macrophages; these are microvilli and membrane ruffles. Since LDL and βVLDL overlap with respect to receptor recognition, it was of interest to determine if the LDL receptor co-localized with the βVLDL receptors. This question has been addressed through 3-D IVEM of whole mount cells which had b een processed for either ligand-gold or immunogold (receptor) chemistry.
A pre-embedding double staining procedure used to observe the effect of fixation on the activity of intercellular adhesion molecule on T and B cells
Authors: Ramberg Jane E. , Tohma Shigeto , Lipsky Peter E.
Published: June 2020

Intercellular adhesion molecule (ICAM-1) appears to be a ligand for LFA-1 dependent adhesion in T cell mediated cytotoxcity. It is found on cells of both hematopoietic and non-hematopoietic origin. While observing the activity of ICAM-1 on the surfaces of interacting T and B cells, we found that we could successfully carry out a pre-embedding double staining procedure utilizing both colloidal gold and peroxidase conjugated reagents.

On 24-well microtiter plates, mitomycin-treated T4 cells were stimulated with 64.1 (anti-CD3) for one hour before the addition, in some instances, of B cells. Following a 12-48 hour incubation at 38°C, the cells were washed and then immunostained with a colloidal gold conjugated RFB-4 (anti-CD22); biotinylated R6.5 (anti-ICAM-1); followed by streptavidin/peroxidase. This method allowed us to observe two different antigens without concern about possible cross-reaction of reagents. Because we suspected ICAM-1 and R6.5 were sensitive to fixation, we tried varying concentrations of fresh paraformaldehyde before R6.5, after R6.5 and after streptavidin/peroxidase. All immunostaining and washing was done on ice with ice cold reagents.
Electron microscope study of the secretory activity of epithelial cells in embryonic thymus
Authors: Alasam H.
Published: June 2020

The possibility that intrathymic T-cell differentiation involves stem cell-lymphoid interactions in embryos led us to study the ultrastructure of epithelial cell in normal embryonic thymus. Studies in adult thymus showed that it produces several peptides that induce T-cell differentiation. Several of them have been chemically characterized, such as thymosin α 1, thymopoietin, thymic humoral factor or the serum thymic factor. It was suggested that most of these factors are secreted by populations of A and B-epithelial cells.

Embryonic materials were obtained from inbred matings of Swiss Albino mice. Thymuses were disected from embryos 17 days old and prepared for transmission electron microscopy. Our studies showed that embryonic thymus at this stage contains undifferentiated and differentiated epithelial cells, large lymphoblasts, medium and few small lymphocytes (Fig. 5). No differences were found between cortical and medullary epithelial cells, in contrast to the findings of Van Vlietet al,. Epithelial cells were mostly of the A-type with low electron density in both cytoplasm and nucleus. However few B-type with high electron density were also found (Fig. 7).
Electron Microscopic study of human herpesvirus-6 (HHV-6)
Authors: Yoshida Mariko , Uno Fumio , Yamanishi Koichi , Ikuta Kazuyoshi , Kurata Takeshi , Nii Shiro
Published: June 2020

Human lymphocytes from cord blood and MT-4 cells were infected with the Hashimoto strain of HHV-6 isolated from a patient with exanthem subitum or with the Z29 strain of this virus isolated from a patient with AIDS, and the infected cells were examined by transmission and scanning electron microscopy. The following observations, characteristic for infection with HHV-6, were obtained. In the nucleus, most of the capsids with a diameter of about 100nm had cores of low density, which seemed to consist of punctate and/or filamentous structures. Tubular structures formed due to some mistakes in the viral assembly were also detected. The most striking characteristic of the ultra structure of this virus was the distinct coating of intracytop1asmic and extra cellular nucleocapsids with a tegument of moderate electron density. These particles coated with a tegument acquired their envelopes by budding at the vacuolar membrane in the cytoplasm. Enveloped viral particles with a diameter of about 170-200 nm were observed in cytoplasmic vacuoles and on the cell surface membrane (Fig. 1).
Neonatal adrenal cortex: Intercellular permeability to the nickel nitrate
Authors: Piezzi Ramón , Gutiérrez Luis , Fogal Teresa
Published: June 2020

The adrenal gland of the rat shows gradual changes during its growth and differentiation. During the first days of life there is a decrease of adrenal weight and cortical volume. During this period the cell-to-cell attachments of the cortical tissue and the permeability of the intercellular spaces to the markers during that period has not received special attention. The purpose of this work is to describe in the neonatal cortex: 1) The permeability of the intercellular spaces to the nickel nitrate used as marker, and 2) Some structural observations of the zone of cellular adhesion between cortical cells. The marker fills the lumen of microcanaliculi. With this purpose the following procedure are made: Adrenal gland of twenty Wistar rats of 1,4,10 and 90 days of age are used. In the day of sacrifice both glands are dissected after perfusion with nickel-glutaraldehyde via cannulation of cardiac left ventricle. The nickel-glutaraldehyde is prepared by dissolving 1 g of nickel nitrate in 0.1M cacodylate buffer pH 7.2. After perfusion, fragments of the glands are refixed in 1% osmium K-ferrocyanide and processed by TEM.
Surface morphology of Purkinje cells and myocardial cells following chemical digestion
Authors: Morita Tetsuo , Shimada Tatsuo
Published: June 2020

From light and transmission electron microscopic studies, it has been long known that Purkinje cells of mammalian hearts have morphological characteristics different from ordinary myocardial cells. In the present study, not only Purkinje cells and myocardial cells but also connective tissue sheaths surrounding these cells were investigated by combined scanning electron microscopy(SEM) and chemical digestion.

The moderator band of adult sheep heart was used because it possessed both Purkinje cells and myocardial cells (Fig.1). Tissue blocks were immersed in Karnovsky’s fixative for 3hr or longer. Some fixed tissues were inrrtersed in a 3-5% aqueous solution of NaClO for 1 min to digest the endocardial endothelium, and then were treated with 8N HCl for 30 min at 60°C to remove connection tissue elements. The others were iirmersed in a 2N NaOH or KOH solution for 7 days at room temperature to digest cellular elements. After freeze-cracking in a 40% dimethyl sulfoxide solution, the tissues were washed throughout in a saline solution containing tween 20, placed in a 1% aqueous solution of tannic acid for 2hr and conductive-stained with 1% OsO4 for 2hr.
Differentiation Processes of the Ocellar Retina of the Honeybee (Apis Mellifera L.)
Authors: Anna Pabst Maria
Published: June 2020

In addition to the compound eyes, honeybees have three dorsal ocelli on the vertex of the head. Each ocellus has about 800 elongated photoreceptor cells. They are paired and the distal segment of each pair bears densely packed microvilli forming together a platelike fused rhabdom. Beneath a common cuticular lens a single layer of corneagenous cells is present.

Ultrastructural studies were made of the retina of praepupae, different pupal stages and adult worker bees by thin sections and freeze-etch preparations. In praepupae the ocellar anlage consists of a conical group of epidermal cells that differentiate to photoreceptor cells, glial cells and corneagenous cells. Some photoreceptor cells are already paired and show disarrayed microvilli with circularly ordered filaments inside. In ocelli of 2-day-old pupae, when a retinogenous and a lentinogenous cell layer can be clearly distinguished, cell membranes of the distal part of two photoreceptor cells begin to interdigitate with each other and so start to form the definitive microvilli. At the beginning the microvilli often occupy the whole width of the developing rhabdom (Fig. 1).
Actin filaments may be involved in the morphological changes of the urinary bladder transitional epithelium
Authors: Seguchi Harumichi , Kobayashi Toshihiro , Okada Teruhiko , Xin Zhang Shu , Garcia del Saz Eva
Published: June 2020

In addition to the unique configuration of the luminal plasma membrane(1), the transitional epithelium of the urinary bladder is characterized by the presence of large numbers of microfilaments. These filaments can be divided into two groups: those of 5 nm, and others of 10 nm in diameter(2). In the present study, these filaments were identified by immunohistochemistry and by the HMM binding technique. Quick-freeze, deep-etching replicas were employed to reveal their three-dimensional arrangement.

Light microscopically, the anti-actin immunoreaction was observed in the epithelial layer, as well as in the lamina muscularis mucosae and in the muscular layer. The intensity of the fluorescence in the superficial cells was weaker than in the intermediate and basal cells. The staining pattern of FITC-labeled phalloidin in the urinary bladder was very similar to that of anti-actin. The immunofluorescence of anti-cytokeratin antibody was exclusively observed in the epithelial layer of the urinary bladder. The reaction was evenly distributed throughout the epithelium.
Ultrastructural characteristics of hyaline cells in chick inner ear
Authors: Oesterle E.C. , Cunningham D.E. , Rubel E.W
Published: June 2020

Recent light-microscopic autoradiographic investigations of the chick auditory end organ suggest that several types of supporting cells adjacent to the receptor end organ, i.e., hyaline cells and/or cuboidal cells, may be precursor cells for regenerating hair cells and supporting cells. Little is known regarding these supporting-cell types. The present study investigated ultrastructural properties of hyaline cells with TEM. Inner ears from four 10-day-old chickens were perfused with a mixture of 2% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M cacodylate buffer. After washes in buffer (0.1 M cacodylate), end-organ dissection, osmication, dehydration in alcohol, and Epon embedding, series of semi-thin (1 um) and ultrathin sections (90 nm) were made, stained with uranyl acetate and lead citrate and observed with a Philips EM 410. Sections from high, mid, and low-frequency regions of the papilla were examined.

A light microscopic picture of the inner ear of birds is shown in Figure 1. Hyaline cells are a single layer of cuboidal epithelial cells which rest on the basilar membrane and extend up to the scala media space.
Ultrastructural studies of the relationship between actin filaments and secretory granules
Authors: Holm Nielsen Ellen
Published: June 2020

In secretory cells a dense and complex network of actin filaments is seen in the subplasmalemmal space attached to the cell membrane. During exocytosis this network is undergoing a rearrangement facilitating access of granules to plasma membrane in order that fusion of the membranes can take place. A filamentous network related to secretory granules has been reported, but its structural organization and composition have not been examined, although this network may be important for exocytosis.

Samples of peritoneal mast cells were frozen at -70°C and thawed at 4°C in order to rupture the cells in such a gentle way that the granule membrane is still intact. Unruptured and ruptured cells were fixed in 2% paraformaldehyde and 0.075% glutaraldehyde, dehydrated in ethanol. For TEM (transmission electron microscopy) cells were embedded in Lowicryl K4M at -35°C and for SEM (scanning electron microscopy) they were placed on copper blocks, critical point dried and coated. For immunoelectron microscopy ultrathin sections were incubated with monoclonal anti-actin and colloidal gold labelled IgM. Ruptured cells were also placed on cover glasses, prefixed, and incubated with anti-actin and colloidal gold labelled IgM.
Effects of gamma radiation on muscle ultrastructure and proteinase activity of grass shrimp (Penaeus monodon F.)
Authors: Perng Fang-Shiow , Yang Jui-Sen
Published: June 2020

Effects of irradiation on ultrastructural changes and proteinase activity of grass shrimp during storage was investigated. Shrimps were irradiated by a 60Co radiation source (total radioactivity 143 KCi and irradiation dose rate 3.0 KGy/hr) at a dose of 40 KGy at frozen temperature. Tissue pieces from the first abdominal segment of the shrimps were primarily fixed in 4 % glutaraldehyde (in 0.1 M K-phosphate buffer containing 3.04 % NaCl and 0.02 % CaCl2, pH 7.2), postfixed in 1 % OsO4, and dehydrated in acetone series and embedded in Spurr’s resin. Sections were stained with uranyl acetate and lead citrate and examined in a Hitachi H-600 transmission electron microscope operated at 75 KV.

Small pieces of muscle were dissected from living grass shrimps and immediately incubated with 5 ml of shrimp body fluid for 30 minutes. The body fluid was released from irradiated whole shrimps after storage at ambient temperature for three weeks. The muscle pieces were also incubated with crude proteinase extracted from irradiated hepatopancreas dissected from shrimps or proteinase (EC 3.4.4.16) from Bacillus licheniformis. After incubation the ultrastrucutre of the muscles was prepared for examination by electron microscopy.
Ultrastructure of the miracidium of Schistosomia japonicum
Authors: Jamuar M. P.
Published: June 2020

Miracidium is the first larval stage emerging frcm the egg produced in the vertebrate host in the life cycle of a Schistosome. They have to find a particular species of snail (the Intermediate host) within few hours for the continuation of the life cycle of the parasite and for which they have to be well equipped. Cilia helps them in the movement and sensory organs aid in locating the host. Some cytolytic substance producing glands helps in their penetration into the host.

Miracidia were obtained from livers of mice infected 2 months previously with cercaria of Schistosoma japonlcum, in the Department of Microbiology, University of Chicago, Chicago, USA. The Miracidia were fixed in freshly prepared 6% Glutaraldehyde in 0.2 M phosphate buffer at pH 7.6 at 4°C for 2 hr. and were post fixed for 1 hr. in 1% Os O4 in the same buffer. They were washed twice with double distilled water and dehydrated in graded series of ethanol.
SEM studies of architecture of thoracic chelate appendages of fresh-water prawn, Macrobrachium lamarrei H.M. Edwards (Decapoda-Palaemonidae)
Authors: Sharma U. D. , Shukla Sanjive
Published: June 2020

Chelate legs of certain crustaceans help in detection and capture, and finally carry the food to the trophic devices. The first and second pair of chelate leges (Periodpods) of M. lamarrei were studied to understand their role in food manipulation. These appendages were detached from the prawn, fixed, and processed. Samples were observed and photographed under an SEM (JEOL JSM 35C).

Six types of bristles/setae have been observed on these podomeres (Figs. 1-11). However, the bristles on podomeres of the second leg are sparsely arranged. The toothed and serrate setae are also missing from the propodite of the second leg. The aboral and adoral surfaces of propodite of I chelate leg have two different types of sensory bristles. The serrate setae are aboral, each with a central shaft (120 μm long) and bilateral projections (15.5. μm long); the adoral ones are toothed setae (297 μm) with sunken base and denticulation at the distal end. A third type, the sawtoothed setae, are on the proximal part of the dactylopodite. These are arranged in 7-8 rows of 4-9 setae each.
An ultrastructural study on the shark liver
Authors: Yamada Masako , Tanuma Yutaka
Published: June 2020

Although many fine structural studies on the vertebrate liver have been reported on mammals, avians, reptiles, amphibians, teleosts and cyclostomes, there are no studies on elasmobranchii liver except one by T. Ito etal. (1962) who studied it on light microscopic level. The purpose of the present study was to as certain the ultrastructural details and cytochemical characteristics of normal elasmobranchii liver and was to compare with the other higher vertebrate ones.

Seventeen Scyliorhinus torazame, one kind of elasmobranchii, were obtained from the fish stock of the Ueno Zoo aquarium, Ueno, Tokyo. The sharks weighing about 300-600g were anesthetized with MS-222 (Sigma), and the livers were fixed by perfusion fixation via the portal vein according to the procedure of Y. Saito et al. (1980) for 10 min. Then the liver tissues were immersed in the same fixative for 2 hours and postfixed with 1% OsO4-solution in 0.1 Mc acodylate buffer for one hour. In order to make sure a phagocytic activity of Kupffer cells, latex particles (0.8 μm in diameter, 0.05mg/100 g b.w.) were injected through the portal vein for one min before fixation. For preservation of lipid droplets in the cytoplasm, a series of these procedure were performed under ice cold temperature until the end of dehydration.
Ultrastructure of vacuolated cells in the liver of rock sole and winter flounder living in contaminated environments
Authors: Stehr Carla
Published: June 2020

Focal and diffuse areas of nonneoplastic vacuolated cells in the liver have been observed with light microscopy in up to 12% of the demersal rock sole (Lepidopsetta bilineata) inhabiting Eagle Harbor, which is a contaminated bay in Puget Sound. A similar type of liver vacuolation has also been observed at higher prevalences in winter flounder (Pseudopleuronectes americanus) inhabiting contaminated areas of Boston Harbor. This paper compares the ultrastructure of vacuolated liver lesions in feral rock sole with that of winter flounder from contaminated areas of Puget Sound and Boston Harbor, respectively.

Rock sole from Eagle Harbor, Puget Sound, and winter flounder from Boston Harbor, were collected by bottom trawl from NOAA research vessels. Fish were killed and immediately necropsied. Adjacent pieces of tissue were collected for light and electron microscopy from grossly visible liver lesions or from the center of the liver. Samples collected for histology were fixed in Dietrich's solution, embedded in paraffin and stained with H&E.
Fine structure of dendritic cells in the epithelial cell layer of the ox tongue
Authors: Kobayashi Kan
Published: June 2020

It is known that some kinds of dendritic cells are distributed in the epithelial cell layer of mucous membrane consisting of stratified squamous epithelium. In the process of exfoliation of the epithelial layer from the underlying connective tissue, dendritic cell bodies exposed on the ruptured surface of the epithelium were detected by scanning electron microscopy. These cells were also observed by light microscopy as well as by transmission electron microscopy.

Dorsal mucous membrane of the ox tongue was fixed in Karnovsky's fixative or in 10% formalin. For scanning electron microscopy samples were immersed in 3N-HCl solution for 2-3 weeks at room temperature. The epithelial cell layer was removed from the underlying connective tissue layer1). They were postfixed in tannic acid and then 1% OsO4 for 1 hr. After dehydration in an ethanol series, the specimens were dried by passing through t-butylalcohol, coated with platinum-palladium and observed under an S-800 scanning electron microscope. For transmission electron microscopy, small pieces of the fixed tissue were post-fixed in 1% OsO4 for 1.5 hr and then embedded in Araldite-Epon.
Organization of the thyroglobulin polysome, studied in Endoplasmic Reticulum surface views of rough endoplasmic reticulum in cultured rat thyroid cells
Authors: Kent Christensen A. , Coon Hayden G.
Published: June 2020

Thyroglobulin is synthesized in the thyroid gland and is subsequently degraded to provide thyroid hormones. Rat thyroglobulin is made up of two identical 330 kD subunits, and the mRNA for each subunit contains about 8,500 nucleotides. Since polysomes have approximately one ribosome for each 90-100 nucleotides of mRNA, a polysome of about 85-95 ribosomes would be expected for thyroglobulin. We have been interested in how this very large polysomes is organized on the membranes of the rough endoplasmic reticulum (RER).

It is well known that bound polysomes assume characteristic shapes on the surface of the RER, resembling beads on a string arranged in circles, spirals, loops, hairpins or other forms. These polysomal shapes can be observed in conventional electron micrographs when the membranes of the RER are seen in surface or en face view, rather than in the usual cross section. Clearcut surface views are infrequent, but the likelihood of seeing them is greatly improved when flattened cells in culture are sectioned in the plane of the cell, since the RER in a flattened cell tends to be oriented in that plane.