bluuebunny/crossref_metadata_2025 huggingface dataset:
Application of Transmission Electron Microscope and Scanning Electron Microscope in experimental tumor studies
Authors: Fengming Gao
Published: June 2020

Transmission electron microscope(TEM) and scanning electron microscope(SEM) were widely used in experimental tumor studies. They are useful for evaluation of cellular transformation in vitro, classification of histological types of tumors and treating effect of tumors. We have obtained some results as follows:

1. Studies on the malignant transformation of mammalian cells in vitro. Syrian golden hamster embryo cells(SGHEC) were transformed in vitro by ThO2 and/or ore dust. In a few days after dust added into medium, some dust crystals were phagocytized. Two weeks later, malignant transformation took place. These cells were of different size, nuclear pleomorphism, numerous ribosomes, increasing of microvilli on cell surface with various length and thickness, and blebs and ruffles(Figs. 1,2). Myelomonocytic leukemic transformation of mouse embryo cells(MEC) was induced in vitro by 3H-TdR. Transformed cells were become round from fusiform. The number of mitochondria and endoplasmic reticulum was reduced, ribosomes and nucleoli increased, shape of nuclei irregular, microvilli increased, and blebs and ruffles appeared(Fig. 3).
Evidences for the phagocytic activity of cultured rat mesangial cells
Authors: Kouda Katsuyoshi , Imada Masato , Igarashi Yoshio , Yamashita Akira
Published: June 2020

It is known that glomerular mesangial cells have two major functions; the control of glomerular blood flow by contraction and the removal of macromolecules from the mesangial region by the endocytosis. As regards the latter function, the evidence of phagocytic activity for particulate materials is controversial. In this study, we attempted to make it clear whether mesangial cells isolated from glomeruli exhibit phagocytic activity in vitro.

Male Sprague-Dawley rats, 6-8 wk old, were used. Glomeruli were isolated from rat renal cortex by the sieving technique according to Morita. The isolated glomeruli were cultured at 37°C in an atmosphere of 95% air, 5% CO2 in a medium containing 5ml of RPMI-1640 with 20% decomplemented fetal calf serum, and subcultured. Mesangial cells purified at the 6-20th succesive cultivations were identified by their morphology and antigenic phenotypes which were analyzed by immunohistochemical method using anti Thy-1, anti Ia, anti leukocyte common(LC) antigen, anti iC3bR, and Mar(anti rat macrophage antigen) 1-4 monoclonal antibodies. Fc receptor was detected with rosette forming method using sheep blood cells(SRBC).
Electron Microscopic study of the parathyroid gland of the melatonin-treated hamster
Authors: Shoumura Shizuko , Emura Shoichi , Yamahira Tomo , Kawada Tomoo , Oda Hiroshi , Arakawa Michio , Isono Hideo
Published: June 2020

Some studies have dealt with the relationship between the pineal gland and the parathyroid gland (PTG) Morphological changes suggest that the pineal gland may inhibit or stimulate the PTG. There are a few light microscopic studies on the effects of pineal extract on the PTG. However, there is no study on the effects of melatonin on the ultrastructure of the PTG. We investigated ultrastructural changes in the PTG of golden hamsters after administration of melatonin.

Materials and methods. Three-month-old female golden hamsters with an average body weight of 130 g were divided into 5 groups of 5 animals each. One group was given 0.2.ml of distilled water as controls. The remaining groups were given 0.2 ml of melatonin solution at a dose of 0.8 mg/100 g body weight. The PTG of each group was removed under pentobarbital anesthesia at 1, 5, 24 and 48 hours after injection. The PTG was immersed in a mixture of 2.5% glutaraldehyde and 2% OsO4, dehydrated through acetone and embedded in Epon 812.
Internalization of gold-labeled IA antigens induces formation of Birbeck granule-like structures in murine Langerhans cells
Authors: Bucana Corazon D. , Munn Gwynneth , Ja Song Min , Dunner Kenneth , Kripke Margaret L.
Published: June 2020

Langerhans cells (LC) are migratory bone marrow-derived dendritic cells that reside in the basal portion of the epidermis. LC are antigen presenting cells; they express MHC Class II antigens, macrophage maturation marker, F4/80, and exhibit a distinctive cytoplasmic pentalaminar organelle called Birbeck granules (BG). Murine LC have fewer BG than human LC. The origin of BG is not clear although several reports suggest that BG are formed by invagination of the cytomembrane to form a membrane sandwich.

In this study, dendritic cells (DC) from the draining lymph nodes (DLN) of mice previously sensitized with fluorescein isothiocyanate were examined by fluorescence and electron microscopy. Expression of la antigen by dendritic cells was determined by immunogold labeling using either Janssen Auroprobe GAM40 (Amersham Corporation, IL) as the secondary antibody or Protein- A-gold (20nm) (Polysciences, Inc., PA) label. Cells were labeled at 4°C and then warmed to ambient temperature. Samples were taken a t various time intervals and processed for TEM. Serial sections of dendritic cells were examined for presence of BG.
Three-dimensional analysis of erythrophagosome in lymph node macrophage by methods of cellular matrix maceration and freeze-replica
Authors: Sasaki Katsunori , Ichikawa Misao
Published: June 2020

Although many ultrastructural works about erythrophagocytosis in various organs have been accumulated, few has been reported in the lymph node. This study discloses the degradative pathways of the erythrophagosomes in the lymph node macrophages ultrastructurally and three-dimensionally by transmission electron microscopy and the methods of cellual matrix maceration and freezereplica.

The portal veins of adult male rats were clamped in the middle with a forceps for 1.5 min after deep anesthesia and laparotomy. At 0.5, 1, 3, 4, 6 hr after the treatment the mesenteric lymph nodes were prepared for cellular matrix maceration, freeze-replica and conventional electron microscopy as follows. CELLULAR MATRIX MACERATION: the lymph nodes fixed with 2% glutaraldehyde for 2 min were postfixed with 1% OsO4 at 4°C for 2 hr, transferred into 70% ethanol, frozen with nitrogen liquid, fractured into two pieces by the blade and macerated in 0.1% OsO4 solution at room temperature for 4 hr.
Fine structural morphology of immunocytes of some molluscs and insects
Authors: Pal S.G. , Baur G. , Ghosh B. , Palit S. , Modak S. , Chowdhury N.
Published: June 2020

In recent years some of the blood cells of several molluscs and insects are characterised as immunocytes. Similar cells from a few invertebrates from India have been looked into under conventional TEM to register the ultrastructural features. This type of study is first of its kind in the subcontinent. Immunocytes from bivalve molluscs Meretrix meretrix, Laroellidens marqinalis and two insect species, apterygote Ctenolepism a longicaudata and pterygote Gesonula punctifrons provide a new set of fine structural information which forms a basis of comparison with those studied earlier.

Immunocytes have been collected from the fresh live species of bivalve molluscs and insects obtained locally at Calcutta. These were fixed in icecold 2% glutaraldehyde in 0.1M phosphate buffer (pH 7.2-7.4) for 1-2 hours at 4-5°C. Subseguently pellets were post-osmicated in 1% OsO4 at room temperature for 1-2 hours. Following dehydration these were embedded in Araldite mixture in plastic capsules and polymerization was effected for 2 days at 60°C. Ultrathin sections were cut in a ultrotome and sections were double stained with Uranyl acetate and lead citrate. These were viewed in a TEM.
Permeability of the pineal gland of the viscacha (Lagostomus maximus maximus) to the lanthanum hydroxide
Authors: Piezzi Ramón , Dominguez Susana , Scardapane Luis , Guzmán Jorge
Published: June 2020

The introduction of lanthanum to study the transport across the capillary endothelium and also as tracer in the skeletal muscle was followed by the use of this salt as an intercellular marker. One of the potential routes for the secretory products of the pineal is the circulatory system. The other postulated route is the ventricular cerebrospinal fluid (CSF). In this sense, a junctional interrelationship between the pineal gland and the CSF has been postulated in different mammals. The present work contains observations on the junctions between the parenchymal cells and the morphological characteristics of the blood vessels of the viscacha pineal using a lanthanum as extracellular marker.

A total of five adult male viscachas (4 to 5 kgs of body weight) are used. The animals are collected near San Luis, Argentina (33°20’ South latitude; 760 m altitude) during summer and winter seasons. On the day of sacrifice the rodents are anesthetized, the brain exposed and a solution of fixative-lanthanum is injected slowly into the third ventricle. The pineal gland is then removed and immersed for 2 h more in the same fixative without lanthanum hydroxide. Ultrathin sections are examined in TEM.
Evaluation of hippocampus in rhesus monkeys after chronic (1-year) inhalation exposure to marijuana: I. Electron microscopy
Authors: Andrews A.M. , Wilson S.W. , Scallet A.C. , Ali S.F. , Bailey J. , Paule M.G. , Slikker W.
Published: June 2020

Exposure of rhesus monkeys (Macaca mulatta) to marijuana via inhalation or to intravenous delta-9-tetrahydrocannabinol (THC), reportedly caused ultrastructural evidence of increased synaptic width. Chronic marijuana smoke in a single rhesus monkey examined after a six month withdrawal time caused ultrastructure changes in the septal, hippocampal and amygdala regions; the synaptic cleft was widened, electron opaque material was found in the cleft and in the pre- and postsynaptic regions, with some clumping of the synaptic vesicles. The objective of our study was to assess neuropathological alterations produced by chronic inhalation of marijuana smoke.

Nineteen male rhesus monkeys, 3-5 years of age and weighing 3-8 kg, were divided into four treatment groups: a) sham control, b) placebo smoke (7 days/ week) c) low dose marijuana (2 times/week with 5 days/week sham) and d) high dose marijuana (7 times/week). A smoke exposure consisted of smoke from one cigarette (2.6% THC) burned down to 10 mm butt length. Smoke was administered via smoke generator (ADL II, Arthur D. Little, Inc. Cambridge, MA) and nose-mouth only masks (local production) equipped with one-way valves.
High-voltage EM description of the types and distribution of sensory endings in the inner conical body of the rat vibrissal follicle-sinus complex
Authors: Mosconi Tony M. , Song Min J. , Rice Frank L.
Published: June 2020

Whiskers or vibrissal follicle-sinus complexes (F-SCs) on the snouts of many mammalian species are structures that have complex, dense sensory innervation. The innervation of F-SCs is remarkably similar in all species with the exception of one site - the inner conical body (ICB). The ICB is an elongated cylindrical structure that encircles the hair shaft near the neck of the follicle. This site has received only cursory attention in ultrastructural studies of the F-SC

Adult rats were perfused after the method of Renehan and Munger2. F-SCs were quartered longitudinally and embedded separately in Epon-Araldite. Serial 0.25 μm sections were cut in either the longitudinal or perpendicular plane through the ICB and examined with an AEI EM7 1.2 MV HVEM (Albany, NY) at 1000 KV. Sensory endings were reconstructed from serial micrographs through at least 20 μm in the longitudinal plane and through 10 μm in the perpendicular plane.

From two to six small superficial vibrissal nerves converge upon the neck of the F-SC and descend into the ICB. The nerves branch into smaller bundles of myelinated and unmyelinated axons along the dorsal side of the hair shaft.
Mast cell-nerve fiber interaction: Ultrastructural studies
Authors: Chi E.Y. , Su M.L. , Tien Y.T. , Henderson W.R.
Published: June 2020

Recent attention has been directed to the interaction of the nerve and immune systems. The neuropeptide substance P, a tachykinnin which is a neurotransmitter in the central and peripheral nervous systems produces tissue swelling, augemntation of intersitial fibrin deposition and leukocyte infiltration after intracutaneous injection. There is a direct correlation reported between the extent of mast cell degranulation at the sites of injection and the tissue swelling or granulocyte infiltration. It has previously been demonstrated that antidromic electrical stimulation of sensory nerves induces degranulation of cutaneous mast cells, cutaneous vasodilation and augmented vascular permeability. Morphological studies have documented a close anatiomical association between mast cells and nonmyelinated nerves, that contain substance P and other neuropeptides. However, the presence of mast cells within nerve fasicles has not been previously examined ultrastructurally. In this study, we examined ultrastructurally the distribution of mast cells in the nerve fiber bundles located in the muscular connective tissue of rat tongues (n=20).
The brain structure and neurosecretory cells of noctuid moth Anadevidia peponis: Noctuidae
Authors: Sato M. , Ogawa Y. , Sasaki M. , Matsuo T.
Published: June 2020

A virgin female of the noctuid moth, a kind of noctuidae that eats cucumis, etc. performs calling at a fixed time of each day, depending on the length of a day. The photoreceptors that induce this calling are located around the neurosecretory cells (NSC) in the central portion of the protocerebrum. Besides, it is considered that the female’s biological clock is located also in the cerebral lobe. In order to elucidate the calling and the function of the biological clock, it is necessary to clarify the basic structure of the brain. The observation results of 12 or 30 day-old noctuid moths showed that their brains are basically composed of an outer and an inner portion-neural lamella (about 2.5 μm) of collagen fibril and perineurium cells. Furthermore, nerve cells surround the cerebral lobes, in which NSCs, mushroom bodies, and central nerve cells, etc. are observed. The NSCs are large-sized (20 to 30 μm dia.) cells, which are located in the pons intercerebralis of the head section and at the rear of the mushroom body (two each on the right and left). Furthermore, the cells were classified into two types: one having many free ribosoms 15 to 20 nm in dia. and the other having granules 150 to 350 nm in dia. (Fig. 1).
Bile canalicular microfilament (BCMF) in experimental cholestasis: Freeze-etching study
Authors: Yasuda Hiromoto , Seki Keiko , Tamaki Yoshimi , Yamanaka Masami , Sano Naoyo
Published: June 2020

In this studies, authors tried to clarify the fine structural features of bile canalicular microfilaments(BCMF) closely related to the bile canalicular structures in the experimentally induced rat liver cells.

Male Wistar rats, administrated Phailoidin 50μg/100g. BW., intraperitoneally for 5 days, and sodium Taurolithocolate(TLC), intravenously, for 40 min., 80 min. and 120 min. After minced liver tissues, small pieces were rapidly frozen on copper block cooled with liquid N2, and were freeze-etched in Balzers 400 or JFD 9000 machines. Specimens of replicae as well as thin sections prepared usual thin section method were observed HU.12A electron microscope at 75 to 100KV.

In freeze-etched replica of normal liver, there were moderate number of microfilament(Mf) with trabecular mesh-work. FIG.1.indicates thin and short filaments closely attached to the trabecules and subsequently related to the organellaes. Bundles of intermediate filaments(If) were also encountered closely linked to the Mf. Phailoidin treated liver, there were markedly increase in number of Mf and occasional small vesicular structures. Lumen of canaliculi were dilated with decrease in number or shortening of microvilli(Mv)(FIG.2).
Observation of hydrated muscle by the use of side-entry environmental cell
Authors: Suda Hitoshi , Ishikawa Akira , Fukami Akira
Published: June 2020

In our laboratory, the wet cell microscopy has been developed to observe hydrated materials by using film sealed environmental cell (hydration chamber; abbreviated as EC). We have also reported results of dynamic observations of hydrated biological or non-biological materials such as the biological reaction process of muscle fiber, the chemical reaction process of photographic silver halide crystals and so on. In this study, to prevent completely the evaporation of solvent in the process of setting of sample and the observation, the spacer composing the EC was improved to have the injection function of two solutions (as seen in figure 1) in addition to the water vapour saturated gas circulation system used previously. Such a new injection method may be very useful for the drying sensitive specimen as well as for the action of two solutions.

As far as one observes the specimen with electron microscope, the radiation damage of specimen may be inevitable. To reduce such a damage, some devices are especially necessary in the EC. Before observing “living” biological specimen, however, one must know the critical dosage at which the biological function is lost. In order to estimate the critical value, we performed one experiment by utilizing the function of muscle contraction. Since we can easily judge whether or not “the muscle is living” by observing the contraction phenomen a which cause very dramatic structure change, the muscle fiber is thought to be a very suitable sample for our aim.
Desmosome-like structure between lymphocytes of developing thymus of foetal lamb
Authors: Alsalami Mohammed T.
Published: June 2020

The foetal thymuses of sheep from 40 days to full term gestation were studied. The earliest thymuses that were studied were obtained from foetuses of 40 days gestation. At this stage of development the thymic tissue was formed of cords of condensed epithelial cells. Lymphopoiesis occurred in these thymuses between the thymus of 40 day old foetus and those of 42 days was striking and indicated that the onset of lymphopoiesis in the foetal sheep occurred abruptly. From 42 days onwards there was a rapid increase in the size of the thymus and in the extent of the lymphopoiesis (Fig. 1). By 45 days gestation the distribution of the lymph cells in the thymus had begun to define the cortex and medulla, and the organ was plainly lymphoid.

Apart from lymphoid and epithelial cells, macrophages, erythroblasts and granulocytic myelocytes were sometimes found in the thymus (Fig. 2). Many of die lymphocytes within the cortex were closely attached to one another, the point of attachment resembling tight junctional complexes that normally found between epithelial cells (Fig. 3 and 4).
“Anti-rigor” cross bridges may reflect altered state of actin
Authors: Trombitás Károly , Pollack Gerald H.
Published: June 2020

We reported earlier that in insect flight muscle, myosin heads could form either rigor bridges or “anti-rigor” bridges, with reversed angle. When rigor develops at constant fiber length, the bridge angle is typically 45°—the rigor bridge. When rigor develops isotonically, so that the specimen can shorten, regularly arranged anti-rigor bridges are also observed.

To investigate anti-rigor bridges further, bumblebee-ilight muscle fibers were prepared as described previously. Fresh fiber samples were stretched to 5-15% beyond their natural length, and then chemically skinned by a two-hour soak in a relaxing solution containing 0.5% Triton X-100. The stretched muscles were then alternately transferred and kept for 20 minutes in relaxing and rigor solutions until some of the fibers broke as a result of the developing rigor tension. Samples were then fixed in rigor, and the broken fibers were separated from the intact fibers.

Micrographs of the intact fibers revealed the conventional rigor-bridge angles (Figure. 1). “Chevrons” formed by bridge pairs pointed away from the Z-line in a regular manner.
The early development of antenna and antennal sensilla in the noctuid moth, Agrotis segetum (Insecta: Lepidoptera)
Authors: Hallberg Eric , Hansén Lina
Published: June 2020

The antennal rudiments in lepidopterous insects are present as disks during the larval stage. The tubular double-walled antennal disk is present beneath the larval antenna, and its inner layer gives rise to the adult antenna during the pupal stage. The sensilla develop from a cluster of cells that are derived from one stem cell, which gives rise to both sensory and enveloping cells. During the morphogenesis of the sensillum these cells undergo major transformations, including cell death. In the moth Agrotis segetum the pupal stage lasts about 14 days (temperature, 25°C). The antennae, clearly seen from the exterior, were dissected and fixed according to standard procedures (3 % glutaraldehyde in 0.15 M cacaodylate buffer, followed by 1 % osmiumtetroxide in the same buffer). Pupae from day 1 to day 8, of both sexes were studied.
An Electron Microscopic study of the liver in certain marine teleosts
Authors: Ooi V.E.C. , Ooi Linda S.M. , Ma Shuyi
Published: June 2020

The hepatic parenchyma of five marine teleosts (belonging to five families in the Teleostei) was investiga ted during the summer months by means of light and transmission electron microscopy. The architectural organization of the livers of these teleosts displays remarkable differences with regard to the interrelationship with the pancreatic tissue. In Lates calcarifer and Lutjanus bohar, prominent pancreatic tissue is found in the liver, following the course of portal vein and hepatic artery and surrounding the branches of these vessels. The pancreatic tissue appears less abundant in Mylio macrocephalus than in the preceding species. In Siganus javus and Gymnothorax schismatorhynchus, there are no dispersed intrahepatic pancreas. The liver lobules are defined by the presence of connective tissue strands in Lates and Siganus, but in the other three species there was no precise lobulation of the hepatic tissue.

The microstructure of the livers in these teleosts is basically the same in all cases, although there are considerable variations inultrastructure of their parenchymal cells. The plates of hepatocytes which are usually two or more cells thick is in terrupted by sinusoids lined with fenestrated endothelial cells and Kupffer cells (Figs. 1, 2).
Crustacean olfaction: The daphnid aesthetascs lead to a changed structural concept
Authors: Elofsson Rolf , Walleén Rita
Published: June 2020

Aesthetascs situated on the first antennae of crustaceans are easily recognized and distinguished from other sensilla by their hose-like appearance and thin cuticle. Their internal structure is known from decapod, mysid and isopod, i.e. malacostracan crustaceans. The aesthetasc is characterized in these animals by a large number of sensory neurons below each single aesthetasc. The neurons give rise to - from their dendrites - two cilia. The cilia protrude into the external hose-like portion of the aesthetasc splitting up into many branches. It is generally agreed that the aesthetascs are the structural base for crustacean olfaction and experiments to verify this have been performed mostly on malacostracan species.

The water-flea, Daphnia, is a non-malacostracan species. It has greatly reduced first antennae and externally only nine pairs of aesthetascs remain (Figs 1, 2). The outer morphology agrees with that of malacostracan species. The internal morphology does not. Each of these aesthetascs has only four sensory cells situated deep inside the animal. The sensory cells have only one cilium (Fig. 3) extending unbranched into the external portion of the aesthetasc (Fig. 4).
Ultrastructural and morphometric examination of renomedullary interstitial cells and their lipid granules treated with a prostaglandin synthetase inhibitor
Authors: Seçkin İsmail , Hekimoğlu Nezih , Taşyürekli Mustafa , Yılmazer Selma , Arda Oktay , Şatıroğlu Güngör
Published: June 2020

Experiments points out that renomedullary interstitial cells (RIC) are responsible for antihypertensive function of kidney medulla, but functional roles of lipid granules (LG) of the cell in prostaglandin and renomedullary hormone synthesis are still being discussed. We searched, the relationship in between LG and PGE2 synthesis capacity of RIC in rats induced with a prostaglandin synthetase inhibitor (indomethacin). Three months old 6 Wis-tar rats used in control and experiment groups. Daily doses of 5 mg/kg indomethacin introduced orally to rats in experiment group for 12 days. PGE2 levels were determined in blood samples obtained from left renal vein (Tab.1). Electron microscopic specimens prepared from inner medulla of right kidneys. We caunted LG in 100 RIC of each rat (Tab.1). Frequency of LG groups in control and experiment groups are shown in (Tab.2).

Observations in experiment group; increased cytoplasmic matrix density, mitochondrial swelling, both enlargement of GER and perinuclear cisterna, decreased number of vesicles and nuclei rich in heterochromatin (Fig:2,3,4), points out a declined synthesis and secretion activity of RIC.
Fourier analysis of two structures in opacifying mouse lens
Authors: Vaezy Shahrara , Clark John I. , Clark Judy M.
Published: June 2020

The neonatal mouse lens opacifies at temperatures below the phase separation temperature (Tcat). The opacification starts at the center of the lens, the lens nucleus, and progressively expands to the peripheral regions of the lens, the lens cortex. Tcat is higher in the lens nucleus than in the lens cortex. The opacification is the result of an increase in the size of the fluctuations in the index of refraction. When these fluctuations are smaller than ½ the wavelength of visible light, little light scattering occurs, and the lens is transparent. It is only when the fluctuations in the index of refraction reach dimensions of ½ the wavelength of visible light that significant light scattering takes place. We have observed two distinct structures in the phase separated mouse lens, both of which have the above criterion for opacification.
Substructural tubular network in the matrix of induced peroxisomes of rat hepatocyte
Authors: Makita Takashi , Hirose Haruko
Published: June 2020

In the course of cytochemical study of the localization of Ca+2-ATPase and Mg+2-ATPase on the peroxisomal limiting membrane, and also X-ray microanalysis of accumulated metalic elements in the matrix of peroxisomes, we were awear of the fact that the matrix of peroxisome contained a number of tubular or circular membranous structure. Those structure in matrix were not always clear in matured peroxisomes and likely to be overlooked but rather evident in newly formed peroxisomes which had lighter matrix. This report is to confirm the tubular substructure in the matrix of peroxisomes those which were proliferated in hepatocytes after administration of clofibrate to rats.

Male Wister rats(250-280g) were fed with diet containing 0.5% clofibrate for 3 weeks. Each animal was perfused with 2.5% glutaraldehyde buffered with 0.1M sodium-cacodylate-HCl at pH7.4. Ultrathin sections were observed with H-700(Hitachi) and EM-002A(ISI Akashi).
A rapid method for the direct identification of paramyxovirus in canine CSF by ultracentrifugation and negative staining as a rule out for distemper
Authors: Steffens W.L. , Ard M.B. , Greene C.E. , Jaggy A.
Published: June 2020

Canine distemper is a multisystemic contagious viral disease having a worldwide distribution, a high mortality rate, and significant central neurologic system (CNS) complications. In its systemic manifestations, it is often presumptively diagnosed on the basis of clinical signs and history. Few definitive antemortem diagnostic tests exist, and most are limited to the detection of viral antigen by immunofluorescence techniques on tissues or cytologic specimens or high immunoglobulin levels in CSF (cerebrospinal fluid). Diagnosis of CNS distemper is often unreliable due to the relatively low cell count in CSF (<50 cells/μl) and the binding of blocking immunoglobulins in CSF to cell surfaces. A more reliable and definitive test might be possible utilizing direct morphologic detection of the etiologic agent. Distemper is the canine equivalent of human measles, in that both involve a closely related member of the Paramyxoviridae, both produce mucosal inflammation, and may produce CNS complications. In humans, diagnosis of measles-induced subacute sclerosing panencephalitis is through negative stain identification of whole or incomplete viral particles in patient CSF.
Fine structures of pseudomonas aeruginosa determined by the freeze-substitition method
Authors: Amako Kazunobu , Takade Akemi , Umeda Akiko , Misumi Hiroyasu , Sawae Yoshirow
Published: June 2020

Pseudomonas aeruginosa is an important pathogen in the infection of compromised host. As pathogenic factors the roles of some cell associated structures like flagella, cell surface layer and the production of slime are reported. Therefore for the understanding of the mechanisms of the infection of this bacteria, the exact data on the morphological observations are indispensable. In this paper we described our recent observations on the fine structures of P. aeruginosa determined by the mild fixation method, the freezesubstitution method.
Ultrastructure of periplasmic bodies in gram-negative bacteria
Authors: Nianming Xie , Shaoqing Ding , Luping Wang , Zenglin Yuan , Guolai Zhan , Shuhui Lang
Published: June 2020

Perhaps the data about periplasmic enzymes are obtained through biochemical methods but lack of morphological description. We have proved the existence of periplasmic bodies by electron microscope and described their ultrastructures. We hope this report may draw the attention of biochemists and mrophologists to collaborate on researches in periplasmic enzymes or periplasmic bodies with each other.

One or more independent bodies may be seen in the periplasmic space between outer and inner membranes of Gram-negative bacteria, which we called periplasmic bodies. The periplasmic bodies have been found in seven species of bacteria at least, including the Pseudomonas aeroginosa. Shigella flexneri, Echerichia coli. Yersinia pestis, Campylobacter jejuni, Proteus mirabilis, Clostridium tetani. Vibrio cholerae and Brucella canis.
Effect of UV-B radiation on the C4 plant Zea mays L.
Authors: Santos Isabel , Almeida J. M. , Salema R.
Published: June 2020

The known reduction of stratospheric ozone by some pollutants increased the amount of ultra-violetligth reaching the earth's surface raising concerns about its effects on life, since UV-B (280-320 nm) is absorbed by proteins and affects important plant processes such as photosynthesis, transpiration, leaf expansion, dark respiration and biomass. It was thought of interest to subject a C4 plant to this type of injury to see if both types of cells and chloroplasts in their leaves reacted similarly to the treatment.

Zea mays var. LG12 were germinated in vermiculite and plants developed under a 14 hlight photoperiod till they had three expanded leaves. From this stage one set was used as control and other received UV-B provided by a Philips TL 40/12 lamp in addition to white light giving an irradiance of 900 μE m-2s -1 .
Ultrastructural details of leaf papilla development in the moss Andreaeobryum macrosporum
Authors: Crandall-Stotler Barbara J. , Bozzola John J.
Published: June 2020

Projecting papillae ornament the abaxial leaf surfaces of many mosses, including those of the primitive, ectohydric taxon Andreaeobryum macrosporum Steere & B. Murr.. Such papillae presumably create a network of capillary spaces on the leaf surface proper, through which water can be effectively transported, while providing a water-repellent surface for CO2 uptake at the papillar tips. Although thesuperficial characteristics of such papillae have been recorded frequently with SEM,nothing is knownof their development or ultrastructural organization.

Plants collected on limestone outcrops in the Brooks Range, Alaska in June, 1989, were transferred, intact, to culture dishes of vermiculite, moistened with Hatcher's nutrient solution, and maintainedinan environmental chamber with a 1160 lux, 16 h day: 8 h night light regime at 5 C constant temperature. Shoot apices, 1-2 mm long, were excised from both freshly collected and cultured specimens intoa 1% Triton X solution, 1 h prior to fixation in 2% glutaraldehyde plus 2 % paraformaldehyde in 0.1 MNa-cacodylate buffer, pH 7.2, for 4 h at room temperature.
The ultrastructure of the double membrane-bound bodies and endoplasmic reticulum in serial sections of wheat spot mosaic-affected wheat plants
Authors: Chen M. H. , Hiruki C.
Published: June 2020

Wheat spot mosaic disease was first discovered in southern Alberta, Canada, in 1956. A hitherto unidentified disease-causing agent, transmitted by the eriophyid mite, caused chlorosis, stunting and finally severe necrosis resulting in the death of the affected plants. Double membrane-bound bodies (DMBB), 0.1-0.2 μm in diameter were found to be associated with the disease.

Young tissues of leaf and root from 4-wk-old infected wheat plants were fixed, dehydrated, and embedded in Spurr’s resin. Serial sections were collected on slot copper grids and stained. The thin sections were then examined with a Hitachi H-7000 TEM at 75 kV. The membrane structure of the DMBBs was studied by numbering them individually and tracing along the sections to see any physical connection with endoplasmic reticulum (ER) membranes. For high resolution scanning EM, a modification of Tanaka’s method was used. The specimens were examined with a Hitachi Model S-570 SEM in its high resolution mode at 20 kV.
Ruthenium tetroxide vapor staining of dry specimens for Scanning Electron Microscopy
Authors: Takahashi Gen
Published: June 2020

For non-coating scanning electron microscope study of dry specimens, exposure of specimens to osmium tetroxide vaper was attempted. Ruthenium tetroxide(RuO4) is a volatile far more reactive oxidant than osmium tetroxide. In the present study treatment with RuO4 vaper has been found to be successful in fixing and conductively staining dry or semidry biological or non-biological specimens without any special device.

The method of RuO4 vaper staining

Dry or semidry specimens of appropriate size are placed in a glass container with a small bottle containing RuO4 solution or crystal powder. The glass container is tightly stoppered and the specimens are exposed to the vaper of RuO4 for half or several hours. These procedures are carried out in a draft at room temperature. The specimen surface is progressively darkened,indicating the binding of reduced ruthenium(RuO2) to the specimen by the oxidation-reduction reaction in vaper phase. The reduced ruthenium render the specimen conductive and stable under the intense electron beam during SEM observation without metal coating.
Tracing the use of freshwater naiad shells as prehistoric implements through scanning electron microscopy and x-ray microanalysis
Authors: Kristina Hirst K. , Thompson Robert G. , Walters Katherine S.
Published: June 2020

Many freshwater naiad shells recovered from prehistoric Iowa archaeological sites have edges exhibiting patterns of use-wear. The ethnographic and ethnohistoric records contain accounts of the use of these shells in the removal of kernels from corn cobs. Recently, archaeologists have begun tracing use of stone implements through analysis of microscopic wear patterns and deposition of material such as opal phytoliths. Opal phytoliths are small silica bodies which plants deposit within and between some cells. Corn is among the plants which produce opal phytoliths in abundance. Experimentally created naiad shell implements were used to remove kernels from corn cobs. The deposits left on the edges of these implements were compared to those of prehistoric naiad shells thought to be utilized. Scanning electron microscopy (SEM) and x-ray microanalysis were utilized.

The prehistoric and experimentally created edges were prepared for examination by cleaning and coating with carbon. Examination of the tools was performed with a Hitachi S-570 electron microscope equipped with a Kevex 8000 x-ray microanalyzer.
Microwave fixation of large tissue
Authors: Sakai Toshio , Uchizono Koji
Published: June 2020

Chemical fixation of large biological tissues in electron microscopy has a disadvantage that fixative doesn’t penetrate well into the deeper part of the tissue. My experience suggests that the poor penetration by chemical fixation can be improved by microwave fixation. Temperature raise caused by microwave irradiation is regulated by the cold bath on the floor of the oven. This paper shows that microwave fixation is readily practicable together with conventional fixation. Well-fixed section with an area as large as 1×5mm is observable and the details of morphological structures are preserved quite well in it.

A commercial microwave oven (National Microwave Oven Model NEM-335,2.45GHz 500/180W) was used for the microwave fixation. Irradiation period was controlled by Omuron H5cN-AN. Young mouse kidney and brain were used. The tissue in a glass vial placed in the cold bath was immersed in the mixture of 2% paraformaldehyde and 0.5% glutaraldehyde in phosphate buffer (pH7.4),followed by microwave irradiation for 10 sec. twice.
Silver stains for the light and electron microscopic demonstration of spirochete changes in syphilis and AIDS
Authors: Hanker J.S. , Giammara B.L.
Published: June 2020

Two new silver stains have been developed in our laboratories to stain, and possibly identify without culture or immunocytochemical staining, Gram(-) bacteria. The first is the PATS reaction which positively stains Gram(-) bacteria (Figs. 1, 2) including those like spirochetes which are difficult to culture. Another stain for Gram(-), as well as Gram(+), bacteria is a variation of the Gram stain. Ordinarily to stain Gram(+) bacteria, and not Gram(-) bacteria, the crystal violet stain is removed from Gram(-) microbes by rinsing with alcohol/acetone. If this rinse step is omitted, the crystal violet iodide remains attached to both the (+) and (-) microbes. It can then be rendered insoluble, electron opaque and conductive by treatment with silver methenamine solution under microwave irradiation. This procedure has been found especially useful in certain cases to demonstrate spirochetes; it appears to be a more effective silver stain than the PATS reaction forTreponema pallidum, the microbe causing syphilis.
Electron spectroscopic imaging of isolated myocardial cells prepared by slam-freeze molecular distillation processing
Authors: Rufner Robyn , Stearns Rebecca C. , Hastings Cindy L. , Borzak Steven , Godleski John J.
Published: June 2020

Isolated adult rat ventricular myocytes serve as a well-defined model to study subcellular events associated with cardiac ischemia. Optimal ultrastructural preservation of in vivo and cultured myocardial cells has been approached by cryo-preservation techniques involving rapid freezing and subsequent freeze substitution or freeze-dried cryosections. Alterations in biochemical structure and composition inherent with chemical fixation procedures can be alleviated by molecular distillation drying of cryo-fixed specimens. In this morphological study, the effects of metal-mirror cryofixation and molecular distillation processing of myocardial cells was evaluated using electron spectroscopic imaging.

Myocardial cells were isolated from the ventricles of adult female Sprague-Dawley rats by enzymatic digestion. Briefly, perfused ventricles were minced and incubated with calcium-free Krebs-Henseleit buffer containing 0.05% collagenase and 0.03% hyaluroxidase with 0.002% deoxyribonuclease II and 0.002% trypsin and 1 mM Ca for 10-20 min at 37° with continuous oxygenation and shaking. The tissue was transferred to KH-enzyme buffer plus 2% bovine serum albumin, filtered through Nitex mesh, sedimented by centrifugation (150xg) and then resuspended in 1 mM Ca KH buffer with 2% bovine serum albumin. The yield of viable cells was greater than 70%.
A film recording and image processing system for confocal microscopy
Authors: Ackerman Larry D. , Jansen W. T.
Published: June 2020

Scanning confocal microscopy has developed into a very useful technique for many scientific investigations. However, commercial development has been so rapid that some recent advances in computer graphics and imaging have not been incorporated into the commercial systems. One particular concern was high quality hard copy with alpha-numeric and graphic overlays. A subsystem was developed to provide this output for the BioRad MRC-500/600 confocal imaging system.

A digital film recorder, an Agfa Matrix Procolor was chosen as the principal element of hardware. This compact unit can record an image at a resolution of 4096 horizontal by 3072 vertical pixels at a cost equivalent to popular analog video film recorders. The interface is a standard IEEE 488 GPIB board. It is compatible with various film emulsions such as Kodak Ecktachrome 100 as well as many of the major graphics arts and image processing programs. The second element of hardware in this system is an ATVista 4M image processing board.
Thrombus formation on artificial surfaces: Correlative microscopy
Authors: Lai Quintin J. , Cooper Stuart L. , Albrecht Ralph M.
Published: June 2020

Thrombus formation and embolization are significant problems for blood-contacting biomedical devices. Two major components of thrombi are blood platelets and the plasma protein, fibrinogen. Previous studies have examined interactions of platelets with polymer surfaces, fibrinogen with platelets, and platelets in suspension with spreading platelets attached to surfaces. Correlative microscopic techniques permit light microscopic observations of labeled living platelets, under static or flow conditions, followed by the observation of identical platelets by electron microscopy. Videoenhanced, differential interference contrast (DIC) light microscopy permits high-resolution, real-time imaging of live platelets and their interactions with surfaces. Interference reflection microscopy (IRM) provides information on the focal adhesion of platelets on surfaces. High voltage, transmission electron microscopy (HVEM) allows observation of platelet cytoskeletal structure of whole mount preparations. Low-voltage, high resolution, scanning electron microscopy allows observation of fine surface detail of platelets. Colloidal gold-labeled fibrinogen, used to identify the Gp Ilb/IIIa membrane receptor for fibrinogen, can be detected in all the above microscopies.
The effects of poloxamer coatings on the phagocytic uptake of polystyrene nanoparticles by peritoneal macrophages
Authors: Peltier N.W. , Uhl J. , Brophy L.M. , Daniels C.R. , Steel V.L. , Merisko E.M.
Published: June 2020

The activity of the mononuclear phagocytic system (MPS) is a primary impediment for site-specific delivery of parenterally administered drugs. Following intravenous administration of nanoparticulate drug earners, the carrier system is recognized as foreign and is rapidly removed from the blood primarily by the mononuclear phagocytes of the liver and spleen. The efficiency of this process renders delivery of particulate drug carrier complexes to other sites of action difficult. Recent studies have demonstrated the extent of sequestration of colloidal particles by the MPS can be effectively modulated by altering the surface properties of drug delivery carriers. Hydrophilic coatings grafted to colloidal carriers have been shown to significantly decrease liver deposition in vivo and reduce phagocytic activity of peritoneal macrophages. To further evaluate this phenomenon, murine peritoneal macrophages were incubated for various periods of time with fluorescenated polystyrene particles (PSP) that were either devoid or coated with nonionic surfactants of the Pluronic and Tetronic series, i.e. poloxamers F108 and T908. After incubation, macrophages were fixed and processed for either TEM or analysis using confocal microscopy.
Immunoelectron microscopic study of secretory granules containing atrial natriuretic peptide in human ventricular cardiomyocytes
Authors: Lee Ching-Ping , Lee Ying-Shiung
Published: June 2020

With the morphological description of the secretory granules characteristically occurring in the atrial muscle cells a broad, deep and rapid series of studies has provided unequivocal evidence for a potent natriuretic and vasorelaxant substance, namely atrial natriuretic peptide (ANP), arising in the atrial granules. It now appears that these granules are not specific for the atrium but are only most frequent in adult atrial cells, since they may occur to lesser extent in the extra atrial tissues of some species. Thus, the purpose of the present report was to characterize the ANP-containing granules arising in the ventricular muscle cells in patients with dilated cardiomyopathy and chronic heart failure by means of immunoelectron microscopy.

Electron microscopic immunocytochemical study of left ventricular myocardium obtained from endomyocardial biopsy was carried out in a total of 50 consecutive patients with various etiologies of heart disease. Biopsied specimens were fixed with 2% glutaraldehyde buffered with 0.1 M cacodylate (pH 7.4) at 4°C for 60 min. and postfixed with 1% osmium tetroxide in the same buffer solution.
Immunocytochemical localization of calcitonin gene-related peptide in the rat spinal lamina X
Authors: Nasu Fumio , Inomata Kenichirou
Published: June 2020

The region surrounding the central canal, lamina X, of the spinal cord is thought to receive in formation concerning the noxious stimulation of the periphery (Nahin RL et al. 1983). The present study is to perform the localization of calcitonin gene-related peptide in region surrounding the central canal of rat spinal cord.

Normal adult Wistar rats were anesthetized with sodium pentobarbital and perfused through the left ventricle with a mixture of 0.25% glutaraldehyde, 8% paraformaldehyde and 1% picric acid in 0.1M phosphate buffer, pH 7.4 for several minutes, 0-4°C. After perfusion the spinal cord were removed and immersed in same fixative for 60 min at 0-4°C. The fixed tissues were rinsed in cold 0.1 M phosphate buffer (pH 7.4) containing 8% sucrose and 1% sodium borohydride for 30 min. For light microscopic study, the paraffin sections and Microslicer-sections were immunostained by the peroxidase-antiperoxidase method (Sternberger LA 1979). For electron microscopical study, the ultrath in frozen sections method (Tokuyasu KT 1986)were used
Immunoelectron microscopic localization of 22 KDA protein antigen in the surface layer of Mycobacterium bovis BCG strains
Authors: Lounatmaa K. , Brander E.
Published: June 2020

Recently a paracrystalline surface layer (the S-layer) was observed as the outermost layer of the cell wall of Mycobacterium bovis strains. This crystalline protein layer was found in the Glaxo BCG strains, one Russian strain and three Finnish strains isolated from the patients with BCG complications.

In the thin sectioned and freeze-etched cells the periodic structure was seen in the outermost layer of the cel wall. The sectioned cells (Fig. 1) were bound together by the layer, which was electron dense (S-layer). In the metal shadowed cells, however, the periodicity was clearly seen (Fig. 2). The linear (oblique) arrangement of the subunits in the S-layer was observed and the distance between the rows of the subunits was measured to be about 5.5 nm.

Cryosections, monoclonal antibody M. bovis 4C3/17 antibody (AGEN Bicmedical Limited, Queensland, Australia) and both 5 nm and 10 nm Auroprobe protein A (Janssen, Belgium) were used to study, whether the 22kDa protein (MPB70) could form the S-layer of M. bovis strains studied.