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Destructive lesions were made in the right olfactory bulb of 16 adult opossums. Following postoperative survival periods of 4 to 31 days, the animals were sacrificed and perfused with 10% Formalin. Frozen sections of the brain were cut in either the coronal, horizontal, or sagittal plane and processed by the Fink-Heimer II method. Degenerating axons of olfactory bulb neurons were traced caudally in the ipsilateral lateral olfactory tract (LOT). Small lesions revealed a topographic representation of the olfactory bulb within the LOT. The dorsal, lateral, and ventral parts of the bulb were, respectively, represented in the dorsal, intermediate, and ventral parts of the LOT. Terminal degeneration was observed in the superficial half of the molecular layer ipsilaterally in the following structures: anterior olfactory nucleus, anterior hippocampal rudiment, olfactory tubercle, piriform cortex, ventrolateral frontal neocortex, lateral entorhinal cortex, nucleus of the LOT, and the lateral aspect of the cortical amygdaloid nucleus. No degeneration was observed in the anterior limb of the anterior commissure. Dorsal and lateral parts of the olfactory bulb projected to the anterolateral aspect of the olfactory tubercle, whereas the ventral part projected heavily to the entire tubercle. There was no evidence of topographic projections to other olfactory structures. The observations of the present investigation indicated that the olfactory bulb projections in the opossum, a primitive mammal, are essentially comparable with those of placental mammals.
Anat Rec 1981 Sep
PMID:Central connections of the olfactory bulb in the American opossum (Didelphys virginiana): a light microscopic degeneration study. 730 16

For identification of paraganglia (PG), samples of para-aortic tissue, tissues containing the coeliac-mesenteric ganglion complex, and the hypogastric ganglia were removed from 3- and 33-month-old male Fischer-344 rats and were processed by the formaldehyde-induced fluorescence method for visualization of catecholamines. Small PG containing 5-30 cells per section were found consistently in young animals. In each of six old rats, large PG containing 500-4000 brightly fluorescent cells per section were detected. Cell counting revealed a 13.5 x increase in number of PG cells between 3- and 33-month-old rats. Microspectrofluorimetric quantitation in old rats showed equal amounts of catecholamines in PG cells and in adrenal medullary cells. Most PG were located in samples from the para-aortic area.
Anat Rec 1981 Nov
PMID:Age related increase in catecholamine-containing paraganglia in male Fischer-344 rats. 730 34

Since conventional chemical fixation may extract tissue components and thus alter structural organization, cryofixation was used to reexamine the ultrastructure of three thick basement membranes: lens capsule, Reichert's membrane, and Engelbreth-Holm-Swarm (EHS) tumor matrix, and two thin basement membranes, those of epididymis and semi-niferous tubules. Cryofixation was achieved by slam freezing followed by either freeze substitution in dry acetone containing 1% osmium tetroxide and 0.05% uranyl acetate or freeze drying in a molecular distillation dryer. The results by both procedures demonstrate that thick basement membranes and the lamina densa of thin basement membranes are composed of a network of anastomosing strands referred to as cords. The cords vary in density and distinctiveness, but their thickness averages 3 to 5 nm in every tissue examined. The spaces separating the cords vary within wide limits, but their mean diameter is approximately 15 nm in every case. Two other common features are 1) the presence within the network of a few 1.5-3.0-nm-thick filaments and 2) 4.5-nm-wide sets of parallel lines referred to as double tracks. When these results are compared with those previously described after conventional fixation, no significant difference is observed in either the cord network or the associated filaments and "double tracks." However, in the thin basement membranes processed by cryofixation, the lamina densa is in direct contact with epithelial cells, whereas, after conventional fixation, the lamina densa is separated from the epithelial cells by a pale layer referred to as lamina lucida or lamina rara. Immunogold labeling of three basement membranes after cryofixation and freeze substitution in acetone containing 0.3% glutaraldehyde yields strong reactions for laminin, type IV collagen, and heparan sulfate proteoglycan. Comparison with previous results indicates that conventional formaldehyde fixation adequately preserves laminin and type IV collagen but causes the loss of some proteoglycan. It is concluded that, except for this loss and the absence of lamina lucida in cryofixed thin basement membranes, the morphological and antigenic features obtained after cryofixation are similar to those observed in the past after conventional fixation.
Anat Rec 1993 Feb
PMID:Cryofixation of basement membranes followed by freeze substitution or freeze drying demonstrates that they are composed of a tridimensional network of irregular cords. 842 Mar 89

Haemostatic forceps contaminated with Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus and Aspergillus flavus were exposed to formaldehyde gas for up to 48 hours at different temperatures and relative humidities. After 24 hours at 22 to 24 degrees C and a relative humidity of 73 per cent they were sterilised reliably and completely.
Vet Rec 1997 Apr 26
PMID:Sterilisation of surgical instruments with formaldehyde gas. 915 45

Malignant catarrhal fever (MCF) is traditionally regarded as a disease with a short clinical course, low morbidity and high case fatality rate. Owing to the limitations of the assays used for laboratory diagnosis. It was difficult in characterise the clinical spectrum of sheep-associated MCF, particularly when the cattle recovered from an MCF-like clinical syndrome. Over a period of three years, 11 cattle that survived MCF for up to two-and-a-half years were identified on four premises. A clinical diagnosis of MCF was confirmed by the detection of ovine herpesvirus-2 DNA in peripheral blood leucocytes using a polymerase chain reaction (PCR) assay that detects a specific 238 base-pair fragment of viral genomic DNA. Of the 11 cattle examined, six recovered clinically with the exception of bilateral corneal oedema with stromal keratitis (four animals) and unilateral perforating keratitis (one animal). The 10 animals available for postmortem examination had disseminated subacute to chronic arteriopathy. Recovery was associated with the resolution of the acute lymphoid panarteritis that characterises the acute phase of MCF, and with the development of generalised chronic obliterative arteriosclerosis. Bilateral leucomata were due in part to the focal destruction of corneal endothelium secondary to acute endothelialitis. Formalin-fixed tissues and/or unfixed lymphoid cells from all 11 cattle were positive for sheep-associated MCF by PCR. These observations indicate that recovery and chronic disease are a significant part of the clinical spectrum of MCF and that such cases occur with some frequency in the area studied. The affected cattle remain persistently infected by the putative sheep-associated MCF gammaherpesvirus.
Vet Rec 1997 May 17
PMID:Chronic and recovered cases of sheep-associated malignant catarrhal fever in cattle. 953

Clinical recommendations for analgesics in laboratory rodents are usually derived from basic research. However, animal models of pain often involve withdrawal reflexes evoked by threshold-level stimuli, whereas pain associated with surgery or disease involves injury and inflammation. Moreover, the analgesics used in research tend to be chosen as exemplars of a drug class, without regard for whether the route of administration is practical, whether the drug has useful kinetics or whether the side effects are tolerable. This paper provides data on the efficacy of drugs from four classes, using the formalin test as a model of injury-induced pain. Formalin (50 microliters, 2.5 per cent) was injected subcutaneously into a rat's paw and the behavioural response (lifting or licking of the paw) was recorded. Buprenorphine at 0.1 mg/kg and dipyrone at 200 mg/kg completely suppressed the pain responses. When formalin was injected six hours after buprenorphine or dipyrone, pain scores were 30 per cent of control scores. In the absence of pain and handling, 0.6 mg/kg buprenorphine was lethal to 25 per cent of rats. Locomotor activity was slightly depressed by 300 mg/kg dipyrone. Xylazine at 2 mg/kg suppressed pain responses, but the analgesia had decreased to less than 50 per cent after two hours and the effects were variable thereafter; at 8 mg/kg rats were unresponsive to a strong pinch. Acepromazine at 2.5 mg/kg reduced pain to 20 per cent of control scores and this level of analgesia was maintained for six hours; neuroleptic effects were prominent at 5 mg/kg.
Vet Rec 1997 May 24
PMID:Options for management of acute pain in the rat. 918 12

The data concerning the effects of age on the brainstem are inconsistent, and few works are devoted to the human vestibular nuclear complex. The medial vestibular nucleus (MVN) is the largest nucleus of the vestibular nuclear complex, and it seems to be related mainly to vestibular compensation and vestibulo-ocular reflexes. Eight human brainstems have been used in this work. The specimens were embedded in paraffin, sectioned, and stained by the formaldehyde-thionin technique. Neuron profiles were drawn with a camera lucida at x330. Abercrombie's method was used to estimate the total number of neurons. We used the test of Kolmogorov-Smirnov with the correction of Lilliefors to evaluate the fit of our data to a normal distribution, and a regression analysis was performed to determine if the variation of our data with age was statistically significant. The present study clearly shows that neuronal loss occurs with aging. The total number of neurons decreases with age, from 122,241 +/- 651 cells in a 35-year-old individual to 75,915 +/- 453 cells in an 89-year-old individual. Neuron loss was significant in the caudal and intermediate thirds of the nucleus, whereas the changes in the rostral third were not significant. The nuclear diameter of surviving neurons decreased significantly with age. There is a neuron loss in the MVN that seems to be age-related. It could help explain why elderly people find it hard to compensate for unilateral vestibular deficits. The preservation of neurons in the rostral third could be related to the fact that this area primarily innervates the oculolmotor nuclei; these latter neurons do not decrease in number in other species studied.
Anat Rec 1998 08
PMID:Neuronal loss in human medial vestibular nucleus. 971 81

Since we had subdivided the cell cycle into 11 stages--four for mitosis and seven for the interphase--and since we had experience in detecting DNA in the electron microscope (EN) by the osmium-amine procedure of Cogliati and Gauthier (Compt. Rend. Acad. Sci., 1973;276:3041-3044), we combined the two approaches for the analysis of DNA-containing structures at all stages of the cell cycle. Thin Epon sections of formaldehyde-fixed mouse duodenum were stained by osmium-amine for electron microscopic examination of the stages in the 12.3-hr long cell cycle of mouse duodenal crypt columnar cells. In addition, semi-thin Lowicryl sections of mouse duodenal crypts and cultured rat kidney cells were stained with the DNA-specific Hoechst 33258 dye and examined in the fluorescence microscope. The DNA detected by osmium-amine is in the form of nucleofilaments, seen at high magnification as long rows of 11 nm-wide rings (consisting of stained DNA encircling unstained histones). At all stages of the cycle as well as in nondividing cells, nucleofilaments are of three types: 'free,' 'attached' to chromatin accumulations, and 'compacted' in all chromatin accumulations, the form of dense spirals within. At stage I of the cycle, besides free and attached nucleofilaments, compacted ones are observed in the three heterochromatin forms (peripheral, nucleolus-associated, clumped). Soon after the S phase begins, chromatin 'aggregates' appear, which are small at stage II, mid-sized at stage III, and large at stage IV. Chromatin 'bulges' also appear at stage III and enlarge at stage IV, while heterochromatins disappear. At stage V, aggregates and bulges accrete into 'chromomeres,' a process responsible for the apparent chromosome condensation observed at prophase. The chromomeres gradually line up in rows and, at stage VIa (prometaphase), approach one another within each row and coalesce to build up the metaphase chromosomes which are fully formed at stage VIb (metaphase). Daughter chromosomes arising at stage VII (anaphase) are eventually packed into a chromosomal mass at each pole of the cell. During stage VIII (telophase), the chromosomal mass is split into large chunks. In the course of the G1 phase, the chunks thin out to give rise to irregular 'bands' at stage IX, the bands are then cleaved into central and peripheral fragments at stage X, and finally the central fragments are replaced by free nucleofilaments and clumps at stage XI, while the peripheral fragments are replaced by peripheral heterochromatin. The "nucleoli" at stages I-III are associated with stained heterochromatin but otherwise appear as unstained lucent areas, except for weakly stained patches composed of histone-free DNA filaments. During stage IV, nucleoli lose patches and associated heterochromatin, while weakly lucent, pale vesicles appear within nucleoli and in the nucleoplasm. By the end of substage VIa, nucleoli generally disappear, while pale vesicles persist around the chromosomes appearing at substage VIb. At stages VIII and IX, the vesicles seem to become strongly lucent and, at stages IX and X, they associate and fuse to yield homogeneous lucent areas, the 'prenucleolar bodies,' which include histone-free DNA patches. During stage XI, groups of these bodies associate to give rise to nucleoli. In conclusion, the cell cycle DNA changes can be classified into 4 broad periods (Fig. 6): 1) Stage I is a 2-hr long interphase "pause," during which the stained DNA shows no signs of either chromosome condensation or decondensation, while the overall nuclear pattern is similar to that in nondividing cell nuclei. Nucleoli are fully developed. 2) From stage II to VIa, the "chromosome condensation" period extends over about 7 hr, during which the events are interpreted as follows. Throughout the S phase (stages II-IV), newly-synthesized segments of nucleofilaments approach one another, adhere and thus build aggregates and later bulges on nuclear matrix sites. (ABSTRACT TRUNCATED)
Anat Rec 1998 11
PMID:The eleven stages of the cell cycle, with emphasis on the changes in chromosomes and nucleoli during interphase and mitosis. 981 Dec 21

Using a recently developed fixation technique for parietal cells (Sugai et al., Acta Anat Nippon 1995:70:S79, 1999:74:S101), we have reinvestigated the organization of the cytoplasmic membrane system in the resting stomach by ultra-high-resolution scanning electron microscopy (SEM). Rat gastric mucosae were microwave-fixed in cacodylate buffer [334 milliosmoles/kg H(2)O (mOsm)], to which 1.0% glutaraldehyde and 0.5% formaldehyde were added. Specimens examined by transmission electron microscopy (TEM) of thin sections revealed cytoplasm packed with tubular membranes similar to images detected by rapid-freeze/freeze-substitution fixation which is generally considered to cause minimal structural alterations. To render the cytoplasmic membranes visible by SEM, fixed mucosae were frozen, fractured, and the exposed cytoplasm of parietal cells was macerated by the aldehyde-osmium-DMSO-osmium procedure. With much of the cell matrix and filaments removed, SEM revealed numerous 30-60 nm tubules which formed a meshwork and also small cisternae. The cytoplasmic surface of the tubules was smooth while some cisternal areas had attached polyribosomes. Vesicles or isolated tubules were not found in appropriately macerated parietal cells. The cytoplasmic surface of the intracellular canaliculus was smooth except for round openings representing the bases of macerated microvilli. In favorable sites connections of the tubular membranes to the canaliculi were clearly visible. Stereo pair views were particularly useful to demonstrate these continuities. Connections between these two membrane compartments suggest the probability of rapid membrane transposition.
Anat Rec 2000 01 01
PMID:Scanning EM of resting gastric parietal cells reveals a network of cytoplasmic tubules and cisternae connected to the intracellular canaliculus. 1060 44

Development of the periodontium involves a series of complex steps that result in the formation of root dentine, cementum, bone and fibres of the ligament. These precisely controlled and timed events require the participation of the enamel organ derived epithelial cells of Hertwig's (HRS) and ectomesenchymal cells of the dental follicle. These events involve rapid turnover of the tissues and cells, including disappearance of epithelial cells of HRS. Thus, it seemed likely to us that programmed cell death (apoptosis) may play a role in the development of the periodontium. Fragments of first molars, obtained from 14- and 29-day-old rats, were fixed in glutaraldehyde-formaldehyde and processed for light and electron microscopy. For the TUNEL method for detection of apoptosis, specimens were fixed in 4% formaldehyde and embedded in paraffin. Results confirmed that epithelial cells of HRS maintain a close relationship with the forming dentine root, and that they may become trapped in the dentino-cemental junction. Some of the epithelial cells exhibited ultrastructural features which are consistent with the interpretation that they were undergoing programmed cell death, i.e. apoptosis. Periodontal fibroblast-like cells showed typical images of apoptosis and engulfed apoptotic bodies. TUNEL positive structures were present in all corresponding regions. It seems therefore that apoptosis of epithelial cells of HRS and fibroblast-like cells of the periodontal ligament constitutes an integral part of the developmental process of the tissues of the periodontium.
Anat Rec 2000 02 01
PMID:Apoptosis in the early developing periodontium of rat molars. 1064 61


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