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Semen assessment, fertility and the selection of Hereford bulls for use in AI.

Nineteen young Hereford bulls were used to study the relationship between semen characteristics and fertility in artificial insemination following 15 320 inseminations. Seven measures of sperm motility, morphological abnormalities, the release of hyaluronidase, ATP content and sperm head measurements were examined as predictors of fertility (49-day fixed-interval non-return rate). Two assessments of motility, three categories of abnormal spermatozoa, acrosomal changes and the release of hyaluronidase had predictive power. Multiple regression analysis showed that a combination of sperm motility after dilution in saline, motility after thawing and the proportion of coiled tails and proximal protoplasmic droplets provided the best prediction of fertility and allowed bulls to be ranked in order of observed non-return rate (%) with a Spearman correlation better than +0.80.

Adenosine Triphosphate↗

Modern concepts of treatment and prevention of chemical injuries.

Chemical injuries are commonly encountered following exposure to acids and alkali, including hydrofluoric acid, formic acid, anhydrous ammonia, cement, and phenol. Other specific agents that cause chemical burns include white phosphorus, elemental metals, nitrates, hydrocarbons, and tar. Even though there are more than 65,000 chemicals available on the market, and an estimated 60,000 new chemicals produced each year, the potential deleterious effects of these chemicals on humans are still unknown. The Superfund Amendments and Reauthorization Act contains extensive provisions for emergency planning and the rights of communities to know about toxic chemical releases. Since 1990, the Agency for Toxic Substances and Disease Registry (ATSDR) has maintained an active, state-based Hazardous Substances Emergency Events Surveillance (HSEES) system to describe the public health consequences risked by access to hazardous chemicals. Most chemical agents damage the skin by producing a chemical reaction rather than hyperthermic injury. Although some chemicals produce considerable heat as a result of an exothermic reaction when they come in contact with water, their ability to produce direct chemical changes on the skin accounts for the most skin injury. Specific chemical changes depend on the agent, including acids, alkalis, corrosives, oxidizing and reducing agents, desiccants, vesicants, and protoplasmic poisons. The concentration of toxic agent and duration of its contact primarily determine degree of skin destruction. Hazardous materials (hazmats) are substances that may injure life and damage the environment if improperly handled. HAZMAT accidents are particularly dangerous for responding personnel, who are in danger from the moment of arrival on the scene until containment of the accident. Consequently, the Superfund Amendment and Reauthorization Act mandates community preparedness for dealing with hazmat accidents. Paramedics and members of the hazmat response team (usually firefighters) must work together to identify toxic chemicals and assess hazardous environments. The contingency plan for hazmat management can be divided into two parts: initiation of the site plan and evacuation. In coping with hazmat incidents, two distinct goals must be achieved concomitantly. First, hazmats must be contained, fire and explosions must be extinguished, and the site eventually must be cleaned. Second, those exposed to hazmats must be treated at the scene of contamination as well as in the hospital and rehabilitation setting.

Acids↗

The density and distribution of intramembrane particles in erythrocytes from persons with muscular dystrophies.

Intramembrane particles in the freeze-fractured faces of erythrocyte membranes from persons with several different types of muscular dystrophy are examined. The density of particles in the protoplasmic face was lower than that of controls, although not at a statistically significant level. No altered pattern of aggregation of these particles was observed.

Erythrocyte Membrane↗

Perineuronal glial nets in the rat spinal cord. A Golgi study.

Perineuronal glial nets in the rat spinal cord were studied. According to the density of the nets and the thickness of the processes which form them, different types of nets were demonstrated. In addition to nets originating from microglial cells, nets formed by smooth protoplasmic astrocytes are also described.

Animals↗

Use of an enzyme-linked immunosorbent assay for serodiagnosis of clinical paratuberculosis in goats. Study by western blotting of false-positive reactions.

An enzyme-linked immunosorbent assay (ELISA) was performed for diagnosis of paratuberculosis in goats, using as antigen a protoplasmic extract (PPA-3). The test was developed on the basis of the results obtained with two serum reference pools, positive and negative respectively. To avoid day-to-day variations, dilutions of the positive serum pool were included in each plate to obtain an arbitrary system, transforming absorbance into immunoglobulin (Ig) G anti-Mycobacterium paratuberculosis units. The ELISA was used on sera of two reference groups of animals. One group consisted of 35 goats suspected of being infected with paratuberculosis, which was confirmed by histological findings and isolation of M. paratuberculosis. The negative group consisted of 61 healthy goats from a farm free of paratuberculosis. The test showed a sensitivity of 100% and a specificity of 91.8%. Absorption of sera with a Mycobacterium phlei suspension did not modify either the sensitivity or the specificity of the test. Sera from the negative group were analysed by Western blotting, and four of them recognized two fractions with a molecular weight of 17.3 and 28.1 kDa.

Absorption↗

Use of disinfectants in zoos and game parks.

Disinfection is used in the animal quarters of zoos and game parks as an adjunct to physical cleaning and the removal of potentially contaminated materials. Disinfection is particularly useful in reducing infection risks in young animal nursery facilities, and in routine cleaning operations of animal quarters and feeding utensils. Specific disinfectants may be selected for certain known microbial contaminants following an infectious disease outbreak. For example, premises contaminated by foot and mouth disease virus are usually disinfected with 2% sodium hydroxide (lye). The disinfectants most commonly used in zoo operations usually have a broad spectrum of microbicidal activities, such as o-phenylphenol salts, especially sodium o-phenylphenol. Equally popular for routine cleaning and sanitizing operations in zoos are quaternary ammonium compounds and chlorine bleach (sodium hypochlorite). It is important to remember that many disinfectants are protoplasmic poisons, or may be caustic or corrosive. Animals must usually be excluded from facilities being disinfected and premises should be rinsed thoroughly, after a suitable environmental exposure time to such disinfectants, before animals are allowed to return.

Animal Husbandry↗

Rapid structural changes in nerve fibers and cells associated with their excitation processes.

In a variety of nerve fibers, cells and other excitable tissues, the electric responses to electric stimuli were found to be accompanied by a transient swelling of the tissues. The rising phase of this swelling coincides with that of the electric response. By use of a heat-sensor made of polyvinylidene fluoride film, it was also shown that the electric responses of many types of excitable tissues are accompanied by simultaneous heat production. To elucidate the origin of this swelling and heat production, the process of Ca2+-Na+ exchange in synthetic anionic gel beads and rods was investigated. It is asserted that the observed signs of nerve excitation are manifestations of a rapid structural change of the cortical gel layer of the protoplasm, plasmalemma-ectoplasm complex. The importance of the rapid movement and rearrangement of water molecules in the cortical gel layer in association with excitation processes is emphasized.

Animals↗

Distribution and retention of spermatozoa with acrosomal and nuclear abnormalities in the cow genital tract.

Two experiments were conducted to evaluate the fate of sperm following uterine insemination. In Exp. I, five pairs of Holstein cows were inseminated with egg yolk-Tris extended semen (approximately 1.0 X 10(9) sperm; .5 ml) from five ejaculates from a single bull that had high levels (approximately 70%) of morphologically abnormal sperm. Cows were slaughtered 12 h after insemination. The genital tracts were removed and promptly clamped into defined regions. Sperm were recovered by flushing with 2.9% sodium citrate buffer. Proportions of abnormal sperm in the various regions were compared with those in the inseminate. Sperm numbers were also determined from each region. Regions of the tract varied in number of sperm (P less than .001), proportions of knobbed acrosomes (P less than .001), tapered heads (P less than .001), protoplasmic droplets (P less than .001), tail abnormalities (P less than .029) and total abnormalities (P less than .002). A total of 63.5 +/- 6.4 X 10(6) sperm was recovered. These sperm were distributed throughout the tract as follows: vagina, 91.8%; cervix, 5.4%; uterine horns, 2.7%, and uterotubal junctions-isthmi, .04%. No sperm were recovered from ampullae. Because retrograde movement of sperm from the uterus occurred in Exp. I, we conducted Exp. II to determine the extent of sperm loss from the genital tract following insemination. Three pairs of Holstein cows were inseminated with .42 X 10(9) sperm (.5 ml; egg yolk-Tris extender) from the same bull used in Exp. I (three ejaculates). All discharged mucus and urine was collected for 12 h after insemination for recovery of sperm. Aspirates (approximately 1 ml) of mucus from the vagina were evaluated during the 12-h post-insemination period for numbers of sperm and leucocytes. Sperm were also recovered from the tract following slaughter (approximately 12 h) to determine retention. Overall, 73 +/- 3.7% of inseminated sperm were recovered. Components were: inseminate lost from the genital tract in discharged mucus, 60 +/- 4.6%; lost in urine, .06 +/- .02%; aspirated from the vagina, 4.4 +/- 1%; adhered to equipment, 1.3 +/- .3%, and retained in the genital tract, 6.5 +/- 1.6%. Predicted numbers of sperm contained in discharged mucus 2 h post-insemination were greater (P less than .009) than at subsequent hours.(ABSTRACT TRUNCATED AT 400 WORDS)

Acrosome↗

Freeze-etch visualizations of some elements in the stria vascularis.

A quick-freeze, deep-etch study was made to observe the three-dimensional aspect of both membranes and organelles in the stria vascularis of the guinea pig. Our replicas showed various membrane characteristics of the vesicles, e.g., a clathrin basket on the inner surface membrane, aggregation of membrane particles on the protoplasmic fracture face, and regularly arranged surface protuberances on the outer surface membrane. In the cell processes of marginal cells, there were not only bundles of microtubules but also another type of fibril. Many gap junctions were observed between marginal, intermediate, and basal cells. In some replicas, the inner surfaces of the gap junctions were covered with particle protrusions with some ordered arrangements. The functional roles of these elements in the stria vascularis are discussed.

Animals↗

Ultrastructural pathology of glial brain tumors revisited: a review.

The ultrastructural pathology of primary brain tumors of glial origin is examined. These are divided into two major groups. The first category comprises astrocytoma with the variants: fibrillary, protoplasmic, gemistocytic, and anaplastic. These are biologically aggressive tumors of a relatively high proliferative potential and include a substantial proportion of cases that transform into the most malignant secondary glioblastoma. The second category, comprised of rather benign tumors of a limited proliferative capacity and a reasonable good prognosis, includes such clinico-pathological entities as pilocytic astrocytoma, pleomorphic xanthoastrocytoma, and subependymal giant cell astrocytoma of tuberous sclerosis. There is no ultrastructural feature, however, which makes it possible to discriminate between major subclasses of astrocytes; but secondary glioblastoma cells, while still retaining the stigmata of neoplastic astrocytes, are characterized by nuclei that seem to be more indented, cisterns of the endoplastic reticulum may be distended, and intranuclear pseudoinclusions are frequently observed. Primary glioblastoma, which probably originates de novo, is characterized by poorly differentiated cells with a paucity of subcellular organelles and no obvious features of astrocytic origin. Granular cell tumor also belongs to neoplasms of astrocytic lineage and the hallmark of this entity is a cell characterized by the presence of numerous membrane-bound, electron-dense autophagic vacuoles. Its malignant analogue is the granular cell glioblastoma. Two subtypes of granular cell glioblastoma have been distinguished. The first is characterized by the presence of numerous granular, electron-dense bodies which correspond to autophagic vacuoles. The second type is characterized by numerous electron-dense, amorphous masses within cellular processes. These electron-dense inclusions are virtually indistinguishable from minute Rosenthal fibers. The pilocytic astrocytoma is virtually indistinguishable at the ultrastructural level from fibrillary astrocytomas but cells tend to be more elongated. Besides Rosenthal fibers, two types of distinctive structures are relatively common in pilocytic astrocytomas: eosinophilic hyaline droplets and round granular bodies, which are composed of large aggregates of electron-dense secondary lysosomes or small electron-dense bodies, respectively. Pleomorphic xanthoastrocytoma is characterized by astrocytes surrounded by basal membranes. It belongs to a peculiar category of astrocytic "desmoplastic" brain tumors occurring in younger patients, the common denominator for which is the presence of basal lamina. The last category in this group is subependymal giant cell astrocytoma, a tumor of bivalent (glial and neuronal) differentiation, the cells of which are characterized by the presence of peculiar crystalloids. The hallmark of oligodendroglioma is the presence of concentric arrays of membranes (so-called membrane laminations, whorls, or scrolls). A fragment of the cytoplasm sequestrated within a particular whorl may contain mitochondria, lysosomes, or abundant glycogen granules. Ependymomas are characterized by a florid picture dominated by the presence of microlumina, cilia with basal bodies (blepharoplasts), microvilli, and long, interdigitating intercellular junctions of the zonulae adherentiae type. Ganglioglioma, the last category covered by this review, is a mixed glio-neuronal tumor. While glial cells are indistinguishable from their counterparts encountered elsewhere (mostly pilocytic astrocytes), the ganglion cells are characterized by abundant intracytoplasmic dense-core vesicles, absence of intermediate filaments, and numerous microtubules. Occasionally a close apposition of ganglion cells and Rosenthal fibers is seen. Dense-core vesicles are pleomorphic and ranged in a diameter from small synaptic vesicles to large lysosome-like neurosecretory granules.

Brain Neoplasms↗

An overview of botanical clearing technique.

Clearing techniques are outlined with reference to their action on the chemical constituents of plant tissue. The most general technique would include pretreatment with solvents, dissolution of protoplasm, dissolution of other substances, bleaching, infiltration with a dense fluid, and staining. Extensive chemical changes go on during these steps and may prevent satisfactory clearing, an important example being the discoloration of phenolic compounds. Rational design of clearing methods for the chemically distinct cell types and tissue seems a likely future development.

Alkalies↗

Discriminative staining methods for the nervous system: luxol fast blue--periodic acid-Schiff--hematoxylin triple stain and subsidiary staining methods.

This paper describes a new series of staining methods which can discriminatively demonstrate every structure of the nervous system, including axons and capillaries, in animal and human materials. Methods described in this paper consist of one primary stain, luxol fast blue-periodic acid Schiff-hematoxylin (LPH) and six different subsidiary staining methods. The LPH triple stain can precisely differentiate the following structures: neurons (Nissl bodies, cytoplasm, nuclear membrane and nucleolus), various kinds of nuclei (glia, ependyma, endothelium, leucocyte, connective tissue, etc.), myelin sheaths, neuronal processes (axons and dendrites), reacted glial cell bodies (protoplasmic astrocytes, foamy cells, etc.), blood vessels (arteries, veins and capillaries), meninges, intervening connective tissue, erythrocytes, lipofuscin granules, amyloid bodies, and others. Subsidiary staining methods are also described briefly. Applications are discussed in the context of staining technology and neuromorphological research.

Animals↗

Freeze-fracture replica study of capillary endothelium after embolization in the dog.

The underlying mechanisms that lead to brain edema following ischemic insult have been subject to much debate. In this study, experimental cerebral infarction was produced in 25 dogs by injecting 1 or 2 silicone rubber cylinders through the cervical internal carotid artery. The animals were sacrificed 24 hours after embolization. Freeze-fracture studies were conducted on the plasma membrane of the capillary endothelium from 15 control and 25 ischemic dogs. No definite findings of tight junction opening were made in the ischemic preparations. Pinocytotic vesicles were seen as concave areas on the protoplasmic face (PF) of the plasma membrane and as protrusions on the extracellular face (EF). The average pinocytotic vesicle count per square micron was increased in ischemic animals. On the luminal side, it reached 22.0 +/- 1.2/sq mu in the 50 PF samples and 29.5 +/- 1.3/sq mu in the 50 EF samples in the experimental preparations, as compared to 7.2 +/- 0.5 sq mu in the 50 PF samples and 9.0 +/- 0.6 sq mu in the 50 EF samples in normal cortex. The average area of the vesicles was also enlarged in experimental animals: 4990.7 +/- 798 sq nm in the 50 PF samples and 4762.8 +/- 878 sq nm in the 50 EF samples, as compared to 3567.7 +/- 570 sq nm in the 50 PF samples and 3404.5 +/- 573 sq nm in the 50 EF samples in normal cortex (p greater than 0.01). These results indicate that transcellular transportation by pinocytotic vesicles plays an important role in the increase of capillary permeability observed in an ischemic model.

Animals↗

Histological grading and bromodeoxyuridine labeling index of astrocytomas. Comparative study in a series of 60 cases.

The histological grade and the bromodeoxyuridine (BUdR) labeling index of 60 astrocytomas of "ordinary" cell types (fibrillary, protoplasmic, gemistocytic, and anaplastic astrocytomas and glioblastomas) were compared to determine whether the grading system reflects the proliferative potential of the tumors. The tumor grade was based on the presence or absence of four criteria (nuclear abnormalities, mitosis, necrosis, and vascular endothelial proliferation): Grade 1, no criterion, Grade 2, one criterion, Grade 3, two criteria; and Grade 4, three or four criteria. The BUdR labeling index, or percentage of S-phase cells, was calculated in paraffin-embedded tumor sections after in situ labeling by intraoperative intravenous infusion of BUdR, 200 mg/sq m. Exponential regression analyses showed a positive correlation between the histological grade and labeling index (r = 0.88, p less than 0.001) that was stronger than the correlations between log labeling index and age (r = 0.55, p less than 0.001) and between grade and age (r = 0.55, p less than 0.001). These results indicate that the histological grading system reflects the proliferative potential of "ordinary" astrocytomas.

Adolescent↗

Detection of tumor necrosis factor-alpha protein and messenger RNA in human glial brain tumors: comparison of immunohistochemistry with in situ hybridization using molecular probes.

Tumor necrosis factor-alpha (TNF alpha) protein and messenger (m)RNA distribution was studied in biopsy samples of glial brain tumors, using immunohistochemistry and in situ hybridization with molecular probes, to investigate the role of this cytokine in tumor proliferation and immunological host defense. Focal expression of TNF alpha was detected in four of four glioblastomas, one of two anaplastic astrocytomas, and four of five low-grade astrocytomas, regardless of their subtype or grade of malignancy, but in none of the normal peritumoral brain tissues used as controls. The TNF alpha protein and mRNA were present in reactive astrocytes and protoplasmic tumor cells, confined to areas of leukocyte or T-lymphocyte infiltrating, and less pronounced in tumor cells at the edge of necrosis. Additionally, TNF alpha reactivity was found in infiltrating macrophages and perivascular microglia. Immunohistochemistry and in situ hybridization for TNF alpha showed comparable reaction patterns and numbers of TNF alpha-positive cells, even though the sensitivity of in situ hybridization was significantly higher. Quantitative evaluation of TNF alpha protein, TNF alpha mRNA, and leukocyte infiltration revealed a significant positive correlation between the TNF alpha-positive reactive astrocytes and the number of lymphocytes present in corresponding areas. Together, these data lead to the conclusion that TNF alpha in reactive astrocytes and monocytic cells within tumor areas of high leukocyte infiltration and in tumor cells at the border of necrosis may represent one defense pathway of the immune system against tumor proliferation.

Adolescent↗

Surface antigens of the syphilis spirochete and their potential as virulence determinants.

A unique physical feature of Treponema pallidum, the venereally transmitted agent of human syphilis, is that its outer membrane contains 100-fold less membrane-spanning protein than the outer membranes of typical gram-negative bacteria, a property that has been related to the chronicity of syphilitic infection. These membrane-spanning T. pallidum rare outer membrane proteins, termed TROMPs, represent potential surface-exposed virulence determinants and targets of host immunity. Only recently has the outer membrane of T. pallidum been isolated and its constituent proteins identified. Five proteins of molecular mass 17-, 28-, 31-, 45-, and 65-kDa were outer membrane associated. The 17- and 45-kDa proteins, which are also present in greater amounts with the T. pallidum inner membrane protoplasmic cylinder complex, had been previously characterized lipoproteins and are, therefore, not membrane-spanning but rather membrane-anchored by their lipid moiety. In contrast, the 28-, 31-, and 65-kDa proteins are exclusively associated with the outer membrane. Both the purified native and an Escherichia coli recombinant outer membrane form of the 31-kDa protein, designated Tromp1, exhibit porin activity, thereby confirming the membrane-spanning outer membrane topology of Tromp1. The 28-kDa protein, designated Tromp2, has sequence characteristics in common with membrane-spanning outer membrane proteins and has also been recombinantly expressed in E. coli, where it targets exclusively to the E. coli outer membrane. The 65-kDa protein, designated Tromp3, is present in the least amount relative to Tromps1 and 2. Tromps 1, 2, and 3 were antigenic when tested with serum from infection and immune syphilitic rabbits and humans. These newly identified TROMPs provide a molecular foundation for the future study of syphilis pathogenesis and immunity.

Animals↗

[A Study On The Fine Structure Of Clonorchis Sinensis, A Liver Fluke: II. The Alimentary Tract And The Excretory System]

A morphological study on the ultrastructures of the alimentary tract and the excretory system of Clonorchis sinensis was conducted. The liver flukes were collected from rabbit liver six months after the experimental infection The worms were washed with 0.85% saline solution and immediately moved to cold 2.5% glutaraldehyde in 0.1 M phosphate buffer pH 7.4. The materials were dissected and fixed for two hours. The blocks were post-fixed in 1% osmium tetroxide. The blocks were embedded in Epon 812. Ultra thin sections were cut with Sovall MT-2 ultramicrotome and stained with uranyl acetate and lead citrate. Sections were then observed with Hitachi HS-7S electron microscope. The following results were obtained in a series of observations. The walls of oral cavity and esophagus comprised tegumental syncytium, basement membrane, loose connective tissue, muscular layer and parenchymal cells. The apical surface and the base of the syncytium were covered with a protoplasmic membrane for each forming numerous invaginations. Granular endoplasmic reticulum was developed in the epithelium of the oesophagus. The gastrodermis of Clonorchis sinensis comprised two types of cells in general. The first cell type was numerous one forming a single continuous layer of epithelial cells. Each of the cells had outfolded cytoplasm into the caecal lumen and lamellae along the cell surface. Among the above epithelial cells, no considerable differences in structure reflecting their functional states were identified. The second cell type was less differentiated in nature and lay within the gastrodermis above the basement membrane but not in contact with the caecal lumen, being overlapped by neighboring gastrodermal cells of the type described above. At this portion the gastrodermis seemed to be a pseudostratified epithelium. There were well-developed lamellae along the surface of epithelia of all canals or duct concerning evacuation. The excretory pore was 7.5 micrometer in diameter and dorso-terminally opended. The epithelium of the excretory pore, a syncytial layer, contained many microtubules unlike the other part of tegumental layer of this worm. The epithelium thickness of the excretory pore was very irregular(1.3-5.5 micrometer).

Journal Article↗

[Ultrastructural antigenic localization in Paragonimus iloktsuenensis during developmental stage by immunogold labeling method].

Antigenic localization in Paragonimus iloktsuenensis worm tissues (tegument, intestine and vitelline gland) in different developmental stages of 2 weeks, 3 weeks, 4 weeks, 5 weeks and 33 weeks from albino rats (Sprague-Dawley) infected with P. iloktsuenensis was observed by electron microscopy. These worm tissues of different developmental stage of P. iloktsuenensis was observed on electromicrograph by immunogold labeling method using P. iloktsuenensis infected rat serum of 10 weeks. Antigenic localization was demonstrated as labeling of gold particles in tissues on electronmicrograph. In tegument, gold particles were labeled on tegumental tissue, generally more numerous on secretory granules in tegumental syncytium 2 weeks than those on the other elder developmental stages, but there was a little variation in antigenicity according to individual worm tissue. In general, antigenicity in tegumental tissue was not strong (gold particles: 0.1-5/1 microns 2). In intestine, a large number of gold particles (15-18/1 microns 2) were labeled in intestinal epithelium. Gold particles were concentrated especially on secretory granules in cytoplasm, and gold particles were labeled not only in cytoplasmic protrusions, but also in intestinal luminal contents. Intensity of labeling of gold particles was not correlated with developmental stage of worms. In vitelline gland, a large number of gold particles were labeled on vitelline globules. The gold particles in vitelline globules (8-11/1 microns 2) were concentrated in protoplasm among segmental globules.

Animals↗