Polyglucosan bodies in the brain of the cat

Polyglucosan bodies (PGB) were examined by light and electron microscopy in the brain of cats without neurological signs. PGB in the feline brain were round, basophilic, PAS-positive, filamentous structures. Both morphologically and histochemically, PGB in the cat were identical to Lafora bodies and similar structures found in man and other animals. Most of the bodies were situated in the neuronal processes, but were not found in perikarya. PGB were disseminated throughout the brain, especially in the cerebral cortex, midbrain, cerebellum and medulla oblongata. The occurrence of PGB in the feline brain may represent an ageing phenomenon.     

Muscular sarcocystosis in two arctic foxes (Vulpes lagopus) due to Sarcocystis arctica n. sp.: sarcocyst morphology,molecular characteristics and phylogeny

The arctic fox (Vulpes lagopus) is a critically endangered species in Norway, and therefore, the small population is closely monitored, and most foxes found dead are subjected to necropsy. In two deceased foxes, thin-walled muscular sarcocysts were first detected in histological sections, and numerous sarcocysts were later found in frozen and thawed muscle samples from Fox 1. These sarcocysts measured 1–12?×?0.1–0.25 mm and had closely spaced, short, knob-like protrusions, giving the cysts a serrated outline. Genomic DNA was extracted from eight isolated sarcocysts (Fox 1) and two muscle samples (Fox 2) and subjected to polymerase chain reaction amplification at four loci: the nuclear 18S and 28S ribosomal RNA genes and internal transcribed spacer 1 region and the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). Both foxes were infected by the same Sarcocystis sp., which displayed little or no genetic variation at the three nuclear loci (99.9–100 % identity) and slightly more variation at cox1 (99.4–100 % identity). Sequence comparisons and phylogenetic analyses revealed that this species was distinct from other named Sarcocystis spp. but was closely related to various species using avian intermediate hosts and possibly identical to an unnamed species reported from two American dogs. The species described from the two arctic foxes was named Sarcocystis arctica n. sp.     

Selective inhibitory effects of pregabalin on peripheral C but not A-delta fibers mediated nociception in intact and spinalized rats

Effects of pregabalin (PGB, 20–80 mg/kg i.v. injection) on spinally-organized nociception were investigated in isoflurane-anesthetized intact and spinalized rats. Responses of single deep spinal dorsal horn (DH) (laminae IV–V) nociceptive-specific (NS) neurons receiving peripheral inputs from A-δ and C fibers to repetitive electrical stimulation (intensity: 3–5 mA; frequency: 1 Hz; pulse duration: 1 ms), mechanical/heat stimulation were recorded extracellularly during physiological condition and s.c. bee venom (BV) induced inflammation. PGB significantly inhibited C-fiber mediated spinal NS neurons' late responses including phenomena of wind-up (temporal summation) and after-discharge. However, the antinociceptive effects of PGB on nociception were not observed until 30 min after its administration. In contrast, no significant inhibitory effect of PGB on A-δ fiber mediated early responses was observed during the experiments. Compared with intact rats, the inhibitory effects of PGB upon nociception vanished in the spinalized animals. This suggests that PGB-induced selective antinociceptive effect on C-fiber mediated nociception is mainly central effects involving supraspinal centers via descending inhibitory controls. Furthermore, pre-treatment, but not post-treatment, with PGB (80 mg/kg) markedly inhibited s.c. BV elicited spontaneous neuronal responses, and noxious mechanical/heat stimuli evoked hyperactivities of spinal NS neurons, indicating that PGB has efficacy of pre-emptive analgesia on pathological pain associated with central sensitization.     

Basophilic bodies of skeletal muscle in hypothyroidism: enzyme histochemical and ultrastructural studies

Basophilic bodies of skeletal muscles from two patients with hypothyroidism were examined by enzyme histochemistry and ultrastructural study of ultrathin sections stained with periodic-acid-thiocarbohydrazide-silver proteinate for polysaccharides. Some additional characterizations of basophilic bodies were observed: basophilic bodies were found exclusively in type 1 fiber; basophilic bodies were devoid of myofibrillary adenosine triphosphatase, oxidative enzymes, and phosphorylase; and both fibrillary and granular components of basophilic bodies stained strongly for polysaccharides. The polysaccharide nature of basophilic bodies is in keeping with the previous suggestion that the formation of basophilic bodies in hypothyroid patients is related to an impairment of carbohydrate metabolism. Their selective involvement of type 1 fiber and preferential occurrence at the myotendinous junction remain obscure.     

Differential in vivo pulmonary degradation of prostaglandins E1, B1, and A1.

Immunoreactive (IR) plasma prostaglandin (PG) levels were measured in samples collected simultaneously through catheters placed in the right ventricle and the thoracic aorta in fasting anesthetized dogs. There were significantly greater levels of IRPGB (P less than .01) and IRPGA (P less than .05), but significantly less IRPGE (P less than .01) in the aorta than in the ventricle. During femoral vein infusion of PGE1, PGB1, and PGA1, respectively, PGE1 was approximately 87% metabolized, but PGB1 and PGA1 were not degraded by the lung. There was no measurable increase in IRPGB or IRPGA levels in the thoracic aorta during intravenous PGE1 infusion. It was concluded that in the resting state PGE is actively degraded by the lung; that the lung very effectively degrades PGE1 but does not degrade PGB1 or PGA1 during infusion of these prostaglandins; and that pulmonary metabolism of PGE1 probably does not result in formation and release of PGB or PGA into the arterial circulation. Additionally, the possibility exists that in the resting state PGB and/or PGA are actively secreted by the lung, but the immunoassay methodology used does not permit resolution of this point.     

Comparative studies on thermoresistance of protein G-binding region and antigen determinant region of immunoglobulin G in acidic colostral whey

The kinetics of the denaturation of antigen determinant (AGD) region and protein G-binding (PGB) region of bovine immunoglobulin G (IgG) in acidic colostral whey were studied by single radial Immunodiffusion (SRID) and protein G affinity chromatography (PrGAC) during the same courses of heat treatment. The kinetic and thermodynamic parameters for heat-induced denaturation of AGD and PGB regions of IgG were determined over a temperature range of 69-81°C. The denaturation reactions of both AGD and PGB regions of IgG were best described with apparent reaction order of 1.2, the activation energy values of the denaturation reactions were 159.42 kJ/mol and 235.37 kJ/mol respectively. This work suggested that the AGD region was more heat-labile than the PGB region of IgG in acidic colostral whey, Moreover, the higher values of the constant for the AGD region meant that, once the unfolding of the region started, denaturation would occur more quickly than the PGB region on IgG molecule. All these factors contributed to the fact that generally the IgG contents in the products of bovine colostrum determined by PrGAC would be higher than those by SRID.     

Identification of neurons with acetylcholinesterase and NADPH-diaphorase activities in the centrifugal visual system of the chick

The isthmo-optic nuclei (ION) and ectopic neurons, which constitute the centrifugal visual system (CVS), are thought to be cholinoceptive and nitrergic. However, it is not clear which neurons express these markers, namely the ones that project to the retina rather than in neurons that only participate in a local circuit. Therefore, to characterize the neurochemical patterns of the centrifugal visual system in the post-hatched chick, retinopetal cells of the isthmo-optic nuclei and the ectopic region were identified via immunolabeling for cholera toxin, a neuronal tracer, which has been injected in the ocular globe. Then, double labeled with acetylcholinesterase histochemistry to reveal cholinergic synapses, or NADPH-diaphorase histochemistry as a nitrergic marker. Briefly, acetylcholinesterase activity was present mainly in cholera toxin labeled cell bodies of the isthmo-optic nucleus and the ectopic region indicating that retinal projecting neurons of centrifugal visual system comprise a cholinoceptive pathway. On the other hand, NADPH-diaphorase histochemistry was present in the neuropile and sparse cell bodies inside of the isthmo-optic nucleus and in ectopic neurons which were not cholera toxin positive suggesting their role in an intrinsic circuit of the centrifugal visual system. These data support the idea that these two neurochemical systems are present in distinct neuronal populations in the centrifugal visual system.     

Pregabalin Alters Nociceptive Behavior and Expression Level of P2X3 Receptor in the Spinal Dorsal Horn in a Rat Model Induced by Chronic Compression of the Dorsal Root Ganglion

P2X3 receptors are present in the spinal dorsal horn (SDH) and play an essential role in the regulation of nociception and pain. Pregabalin (PGB) has been used as a new antiepileptic drug in the treatment of neuropathic pain. However, it is unclear whether PGB‐induced analgesia was associated with the P2X3 receptor in SDH. Here, rats were randomly divided into four groups (n = 12 per group), including 2 sham operation groups, which was treated by normal saline (Sham + NS group) or PGB (Sham + PGB group), other 2 groups with chronic compression of the dorsal root ganglion, a normal saline‐treated CCD group (CCD+NS group), and a PGB‐treated CCD group (CCD + PGB group). A rat model of neuropathic pain was used by compressing the right L4 and L5 dorsal root ganglia. Each group was evaluated using the mechanical withdrawal threshold (MWT). The mRNA and protein levels of the P2X3 receptor in the ipsilateral SDH were measured by RT‐PCR, western blot, and immunofluorescence on 14 day after CCD operation. CCD rats showed the highest mechanical hyperalgesia and the lowest pain threshold in the four groups. Simultaneously, CCD rats showed higher P2X3 mRNA and protein expression in ipsilateral side of the SDH than the sham operation rats. However, the MWT was increased and expression of P2X3 mRNA and protein in the ipsilateral SDH in CCD rats was decreased 3 days after PGB treatment. Thus, PGB may partially reverse mechanical hyperalgesia in CCD rats by inhibiting P2X3 receptor expression in the ipsilateral SDH. Anat Rec, 296:1907–1912, 2013. © 2013 Wiley Periodicals, Inc.     

Brain alpha‐amylase: a novel energy regulator important in Alzheimer disease?

Reduced glucose metabolism and formation of polyglucosan bodies (PGB) are, beside amyloid beta plaques and neurofibrillary tangles, well‐known pathological findings associated with Alzheimer''s disease (AD). Since both glucose availability and PGB are regulated by enzymatic degradation of glycogen, we hypothesize that dysfunctional glycogen degradation is a critical event in AD progression. We therefore investigated whether alpha (α)‐amylase, an enzyme known to efficiently degrade polysaccharides in the gastrointestinal tract, is expressed in the hippocampal CA1/subiculum and if the expression is altered in AD patients. Using immunohistochemical staining techniques, we show the presence of the α‐amylase isotypes AMY1A and AMY2A in neuronal dendritic spines, pericytes and astrocytes. Moreover, AD patients showed reduced gene expression of α‐amylase, but conversely increased protein levels of α‐amylase as well as increased activity of the enzyme compared with non‐demented controls. Lastly, we observed increased, albeit not significant, load of periodic acid‐Schiff positive PGB in the brain of AD patients, which correlated with increased α‐amylase activity. These findings show that α‐amylase is expressed and active in the human brain, and suggest the enzyme to be affected, alternatively play a role, in the neurodegenerative Alzheimer''s disease pathology.     

Nitric oxide synthase immunoactivity and NADPH diaphorase enzyme activity in neurons of the gastrointestinal tract of the toad, Bufo marinus.

The presence of nitric oxide synthase (NOS) was demonstrated immunohistochemically, and NADPH diaphorase was demonstrated by enzyme histochemistry in neurons throughout the gastrointestinal tract of the anuran amphibian, Bufo marinus. Successive staining showed that NOS immunoreactivity and NADPH diaphorase activity occurred in precisely the same subgroup of enteric neurons. Subsequent detailed studies of the distribution of these neurons were made using NADPH diaphorase histochemistry. Numerous reactive nerve cell bodies and fibres were found in the myenteric plexus from the esophagus to the cloaca. A dense innervation of the longitudinal and circular muscle layers occurred throughout the gastrointestinal tract. The lamina muscularis mucosae was only prominent in the stomach, where it was sparsely innervated. Reactive nerve cell bodies were common in the submucosa of the large intestine, less common in the small intestine and extremely rare in the stomach and esophagus. Reactive fibres contributed to subepithelial plexuses in the esophagus, colon, rectum and cloaca. It is concluded that NOS/NADPH diaphorase is conserved amongst vertebrate classes and that NO is a likely neurotransmitter in the toad gastrointestinal tract.     

Morphology of the lingual papillae in the raccoon dog and fox

The dorsal lingual surfaces of the raccoon dogs (Nyctereutes procyonoides) and fox (Vulpes vulpes japonica) were examined by scanning electron microscopical (SEM) observations. The distribution and type of the lingual papillae found in the raccoon dog were similar to those in the fox. Filiform, fungiform, foliate and vallate papillae were observed. The filiform papillae were distributed over the entire dosal surface of the tongue. Each filiform papilla on the apical surface of the tongue had several pointed processes. The filiform papillae of the lingual body consisted of a main papilla and some secondary papillae. The fungiform papillae were present rounded bodies, and more densely distributed on the lingual apex. The foliate papillae were seen on the dorsolateral aspect of the tongue. The vallate papillae were located on both sides of the posterior end of the lingual body. Each papilla was surrounded by groove and crescent pad. On the periphery of the papillae, large conical papillae were observed.     

大鼠背海马内生长抑素mRNA神经元的老年变化

采用原位杂交组织化学及图像定量分析法，研究了大鼠背海马中生长抑素（ＳＳ）ｍＲＮＡ神经细胞的老年变化。年轻大鼠内，ＳＳｍＲＮＡ胞体主要分布于ＣＡ１和ＣＡ２区的锥体层和多形层、ＣＡ３区的辐状层和多形层，以及齿状回的多形层。老年大鼠内，ＳＳｍＲＮＡ胞体则主要集中于背海马的多形层。和年轻大鼠相比，老年大鼠背海马内ＳＳｍＲＡＮ胞体数量显著减少，胞体灰度值明显升高，而胞体截面积无明显年龄变化。结果表明，大鼠背     

Bactericidal Activity of Blood of Rabbits Vaccinated With Homologous Antigens of Campylobacter fetus (Vibrio fetus)

Rabbits were vaccinated with the following Campylobacter fetus var. venerealis (Vibrio fetus) antigens: whole-cell (WC), autoclaved (A), boiled (B), and purified postgrowth broth (PGB). Bactericidal activity of freshly drawn heparinized blood against the organism was determined after each vaccination. In all cases bactericidal activity of the blood of vaccinated rabbits was higher than for nonvaccinated rabbits. The in vitro bactericidal activity of the blood was determined in two separate experiments. In experiment I the bactericidal activity of the blood of rabbits vaccinated with PGB antigen was the same as that of rabbits vaccinated with WC antigen and higher than that of rabbits vaccinated with A antigen after the third vaccination. In experiment II the bactericidal activity of blood of rabbits vaccinated with PGB antigen was the same as that of those vaccinated with WC antigen after the second and third vaccinations and higher than for rabbits vaccinated with A antigen after the third vaccination. Blood of rabbits vaccinated with A antigen was less bactericidal than blood of rabbits vaccinated with B antigen after the third vaccination, indicating the presence of a surface antigen destroyed by autoclaving but not by boiling. The in vivo and in vitro whole blood bactericidal tests are more sensitive for measuring the response of rabbits vaccinated with WC, B, A, or PGB antigens than is the plate agglutination test.     

Purification and Partial Characterization of a Vibrio fetus Immunogen

An antigen released into broth culture medium (2 mg/100 ml of culture medium) during 20-hr growth of Vibrio fetus was removed from the growth medium by salt precipitation [50 g of (NH₄)₂SO₄/100 ml of broth] and centrifugation. About 30 mg of the postgrowth broth (PGB) antigen protein was removed from other salt precipitable material during one polyacrylamide gel electrophoretic run. The PGB antigen was further purified by using gel filtration on Sephadex G-200, and a molecular weight of 135,000 was established. The purified PGB antigen was shown to contain protein and carbohydrate but not lipid and was a glycoprotein. The antigen had an isoelectric point at pH 4.2 and an R_F value of 0.30 on acrylamide gel electrophoresis. At least one PGB antigen was detected in all (31) V. fetus isolants tested. These antigens were heat labile.     

一氧化氮合酶和5-羟色胺在体外培养的胚鼠脑干神经元内的共存研究

采用 NADPH-d组织化学和 5 -羟色胺免疫组织化学双重反应技术 ,探讨在体外培养条件下 E14 d SD胚鼠脑干神经元中5 -羟色胺与一氧化氮合酶阳性神经元的体外生长发育过程以及 5 -羟色胺与一氧化氮合酶的共存。将 E14 d SD胚鼠脑干细胞悬液接种于 2 4孔培养板 ,实验按培养龄分 4组分别在接种后 5 d、10 d、15 d和 2 1d终止培养 ,先进行 NADPH-d组织化学反应 ,再行 5 -羟色胺免疫组织化学反应。结果显示 :各实验组均可观察到三种神经元 ,即 5 -羟色胺免疫阳性神经元、一氧化氮合酶阳性神经元和两者的双重阳性神经元 ,其中双重阳性神经元占多数 ,并以梭形神经元为主 ,胞体呈三角形、多角形、梭形或卵圆形。 10 d组的双重阳性神经元突起最长可达 60 0μm以上 ,末端可见明显的生长锥 ,轴突终末分支上分布有大量的膨体 ,并与其它神经元的胞体或突起之间形成接触。本研究结果证明一氧化氮合酶和 5 -羟色胺可以在体外培养的胚鼠脑干神经元内共存 ,提示一氧化氮合酶可能与 5 -羟色胺共同参与胚脑的发育。     

大鼠创伤性脑损伤后星形胶质细胞的变化

目的：探讨大鼠创伤性脑损伤后星形胶质细胞的形态学变化及GFAP和NOS的表达情况。方法：采用大鼠自由落体脑损伤模型，伤后1、3、7d取脑切片，行Nissl染色以及GFAP免疫组化和NADPH—d组化单标记及双标记染色。结果：损伤区周围皮质GFAP阳性细胞胞体增大、突起增粗增长，GFAP阳性细胞数量与正常侧及对照组相比，伤后1d即有明显增加，伤后3d、7d数量持续增加；损伤侧海马CAI～3区和DG各层GFAP阳性细胞排列紊乱，胞体增大、突起增粗增长，GFAP阳性细胞数量与正常侧及对照组相比则无明显变化。损伤区周围皮质、损伤侧海马NOS阳性细胞数量明显增加。伤后3d损伤区周围皮质和损伤侧海马中GFAP与NOS双标细胞分别占GFAP阳性细胞的14．2％和13．4％左右。结论：大鼠创伤性脑损伤后大量的星形胶质细胞活化、GFAP表达增加并且部分转化为NOS阳性细胞，提示其参与了脑组织的损伤与修复过程。     

Innervation of rabbit fetal lungs

Nerve fibers, autonomic ganglia, and neuroepithelial bodies of the lungs of rabbit fetuses, 17 to 31 days gestational age, were studied with neurohistological techniques including silver impregnation, acetylcholinesterase histochemistry, and glyoxylic-1acid-induced histofluorescence for monoamines. The silver impregnation method showed that nerve fibers and ganglia accompanied the bronchi and large pulmonary blood vessels to enter the developing lungs by the 17th day of gestation. Cholinergic and adrenergic nerves began to appear in the walls of the bronchi on the 21st day. The developing pulmonary arteries had accompanying adrenergic nerves on the 25th day. Acetylcholinesterase-positive parasympathetic ganglia were seen on the 27th day. Silver-impregnated nerve fibers in the developing alveolar walls and pleura were found on the 25th day. Neuroepithelial bodies and specialized single cells which were argyrophilic, acetylcholinesterase-positive, and fluorescent could be demonstrated in 19–21-day-old and older fetuses; and some of these structures were innervated by sensory and autonomic motor fibers. These observations indicated that nervous tissue and neuroepithelial bodies appeared in the lungs during the glandular stage of the lung development and that differentiation of adrenergic and cholinergic nerves began in the late glandular stage.     

Expression of tyrosine hydroxylase-like immunoreactivity in cell bodies in intraocular spinal cord grafts: a comparison with classical Falck-Hillarp histochemistry

Earlier classical monoamine fluorescence histochemical and immunohistochemical studies describing the distribution of catecholamine neurons in the central nervous system led to the conclusion that all catecholamine fibre systems in the spinal cord have a supra-spinal origin. However, recent reports describing catecholamine neurons in upper cervical segments and at the lumbosacral junction have modified this picture. Isolated intraocular grafts of spinal cord offer a possible model with which to evaluate the presence of intrinsic monoamine neurons. Using immunohistochemistry with antiserum to tyrosine hydroxylase (TH) and Falck-Hillarp fluorescence histochemistry, such grafts from lower cervical segments were found to contain TH-immunoreactive perikarya. However, it was not possible to visualize cell bodies in grafts processed for fluorescence histochemistry. These findings raise questions about the sensitivity of the monoamine fluorescence histochemical method, and about the specificity of TH-immunohistochemistry. Perhaps TH-positive, catecholamine-negative neurons are more common than previously held or alternatively, TH expression in non-catecholamine neurons is induced by perturbations such as grafting brain tissue.     

So-called sclerosing hemangioma of the lung--two cases report with ultrastructural study

Sclerosing hemangiomas of the lung are benign neoplasms of uncertain histogenesis. We analysed two cases of sclerosing hemangiomas of the lung with histochemistry and electron microscopy. They had a variegated histologic appearance characterized by an admixture of solid, hemorrhagic, papillary and sclerotic lesions. Characteristic uniform round cells, unique to this tumor, were found within the stroma in all lesions. In the electron microscopic examination, we found Weibel-Palade bodies like small bodies in the tumor cells. We suspect hypothesis originating in the endothelial cell can not be completely excluded yet. Sclerosing hemangioma is a distinct clinicopathologic entity and should be distinguished from other benign neoplasms or inflammatory lesions of the lung.     

Comparative behavioral effects between synthetic 2,4,5-trimethylthiazoline (TMT) and the odor of natural fox (Vulpes vulpes) feces in mice

Synthetic 2,4,5-trimethylthiazoline (TMT)--a component of red fox (Vulpes vulpes) feces--is frequently used to induce unconditioned fear in rodents. Surprisingly, direct comparison between TMT and natural fox feces odor is almost nonexistent. In this study, Experiment 1 compared the avoidance in relation to TMT concentration, natural fox feces, and gender of fox and mice. Results show that the avoidance is (a) higher with either pure or 50% TMT as compared to natural fox feces, whereas the difference is slight with 10% TMT, and (b) significantly higher for the female mouse group compared to the male mouse group with TMT as well as natural fox feces. In addition, no clear difference in effect was observed between male and female fox feces. Experiment 2 compared behavioral parameters recorded as an index of fear and anxiety, general activity, and avoidance in elevated plus-maze and open-field chamber between 10% TMT and natural fox feces in relation to the estrus cycle of the mice. Results show no cycle period effect--except for the avoidance parameter "distance to odorant"--and no different effects between 10% TMT and natural fox feces except for freezing.