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PubMed1976

Histochemical changes effected by aldosterone on compensatory renal growth in the rat.

The activity of succinic dehydrogenase, adenosine triphosphatase, alkaline phosphatase and cytochrome oxidase was studied histochemically in the remaining kidney of male albino rats which had been subjected to unilateral nephrectomy and were subsequently treated with aldosterone. A marked increase in the activity of succinic dehydrogenase, adenosine triphosphatase and alkaline phosphatase was noted on the initial postoperative days. The increase noted in succinic dehydrogenase activity is considered particularly important because in the control group succinic dehydrogenase activity remained unchanged throughout the experiment.

Adenosine Triphosphatases↗
PubMed1977

Histophysiological studies of prostaglandin F2alpha on isolated organs. I. Effect of prostaglandin F2alpha on the heart.

The physiological and histochemical effects of PGF2alpha on isolated rabbit hearts were examined. The results showed a positive inotropic effect. The coronary flow increased. From the histochemical studies, adenosine triphosphatase (ATP-ase) and succinic dehydrogenase activities were increased while that of alkaline phosphatase was decreased. Glycogen granules were depleted. These findings were discussed on a histophysiological basis.

Adenosine Triphosphatases↗
PubMed1999

Detection of activated eosinophils in nasal polyps of an aspirin-induced asthma patient.

Aspirin-induced asthma (AIA) is frequently accompanied by nasal polyps. Eosinophil infiltration is a characteristic feature of nasal polyps associated with AIA. Even though steroids are well known to be effective on managing AIA and its nasal polyps, histochemical examinations after steroid therapy and at recurrence, involving eosinophil infiltration of nasal polyps, have been less studied. To know the histochemical effects of steroid treatment on eosinophil accumulation in nasal polyps of AIA and the histochemical feature of a recurring polyp and to detect distributional differences between storage and secreted forms of eosinophil cationic proteins, we carried out immunocytochemical labelling with antibodies against EGI (recognizing resting and activated eosinophils) and EG2 (recognizing only activated eosinophils), and determined eosinophil infiltration in nasal polyps that were obtained before and after steroid treatment, and at recurrence of polyps. A large number of eosinophils in AIA polyps were found before steroid treatment and at recurrence, and they were predominantly composed of activated eosinophils (EG2-positive). In contrast, eosinophil infiltration was rare in polyps obtained immediately after steroid treatment. This finding suggests that eosinophil infiltration may be associated with nasal polyp formation in AIA, and that activation of eosinophils plays an important role in accumulation of eosinophils and polyp formation beginning with the initial stage.

Adrenal Cortex Hormones↗
PubMedDate not supplied

Some histochemical and electrophysiological effects of isatin.

Qualitative and semiquantitative histochemical investigations of acetylcholinesterase after a single or repeated administration of isatin, 160 mg/kg ip once or daily for 8 consecutive days have revealed a significant decrease of the histochemical reaction for acetylcholinesterase in chronically treated rats. The cholinergic effect of isatin was tested in the electrophysiological experiment after a single dose of 160 mg/kg isatin and was partially antagonized by atropine.

Acetylcholinesterase↗
PubMed2005

Effects of maternal deprivation on the adrenocorticotrophic and gonadotrophic axes in the hypothalamo-pituitary unit of preweanling female sheep: the histomorphometric approach.

This study was designed to investigate the histochemical effects of maternal deprivation on the adrenocorticotrophic and gonadotrophic axes in the hypothalamo-pituitary unit of preweanling lambs. Twelve-week-old female lambs were divided into either the control (lambs reared under undisturbed maternal conditions; n=3) or the maternally deprived group (lambs separated for three days from their dams; n=3). The corticotrophin-releasing hormone (CRH) and gonadotrophin-releasing hormone (GnRH) in the median eminence and the adenohypophyseal adrenocorticotrophin (ACTH), gonadotrophins (LH and FSH) and mRNAs for their beta-subunits were investigated using the immunohistochemistry or hybridohistochemistry. In maternally deprived lambs, the percentage of the area occupied by immunoreactive (ir)-CRH nerve terminals was lower (P<0.05) and the percentage of the adenohypophyseal area (PAA) occupied by ir-ACTH cells was higher (P<0.05) compared with the control lambs. In the hypothalamo-gonadotrophic axis of maternally deprived lambs the percentage of area occupied by ir-GnRH nerve terminals was higher (P<0.05) and the PAA occupied by ir-FSHbeta cells was lower (P<0.05) in comparison with controls. The PAA occupied by gonadotrophs detected using hybridohistochemistry was higher (P<0.05) for LHbeta-mRNA in contrast to a lower (P<0.05) percentage for FSHbeta-mRNA in maternally deprived lambs compared with those staying with dams. In conclusion, maternal deprivation affected the accumulation of CRH and ACTH. The different and more striking alterations in FSH synthesis and storage in comparison with those concerning LH were observed in maternally deprived lambs. Thus, rupture of the preweanling young-mother social contact can affect the gonadotroph population activity, especially that relating to FSH-producing cells in the infantile female sheep.

Adrenocorticotropic Hormone↗
PubMed2003

Local intracerebral delivery of endogenous inhibitors by osmotic minipumps effectively suppresses glioma growth in vivo.

The systemic administration of endogenous inhibitors significantly reduced the growth of human glioma in vivo, but required the production of a large amount of biologically active protein. In this study we reduced the amount of protein needed and optimized the therapeutical response by delivering the endogenous inhibitors locally into the brain by osmotic minipumps. Human hemopexin fragment of MMP-2 or COOH-terminal fragment of platelet factor-4 were delivered locally and continuously into the brain of mice implanted intracranially with glioma cells, by osmotic minipumps connected to an intracranial catheter. Local delivery of human hemopexin fragment of MMP-2 and COOH-terminal fragment of platelet factor-4 significantly inhibited the growth of well-established malignant glioma in nude and BALB/C mice. When the inhibitors were given at the same concentration, the efficacy of the local delivery was much higher than that reached with the systemic administration, both when the inhibitor was administered daily or continuously by s.c. minipumps. Moreover, the local delivery reduced the amount of protein needed to reach a significant therapeutic response. Intracerebral delivery maintained a long-term control of glioma growth and inhibited glioma recurrence in a surgical resection model. Treatment showed no side effects. Histochemical analysis of tumors showed that the tumor growth inhibition was the result of a decrease in tumor vasculature and a change in tumor vessel morphology. Our data demonstrate that local intracerebral delivery of endogenous inhibitors effectively inhibits malignant glioma growth and reduces the amount of protein needed to reach a therapeutical response.

Amino Acid Sequence↗
PubMed1988

Effect of collidine (2,4,6-trimethylpyridine) on rat pineal gland and vas deferens nerves. Further evidence for a monoamine-releasing effect.

In previous work of our laboratory it was demonstrated that collidine (2,4,6-trimethylpyridine) abolishes the core osmiophilia and chromaffin reaction from rat pinal gland and vas deferens nerves. This abolition was apparent when tissues were briefly incubated in collidine or when they wer fixed in glutaraldehyde or osmium tetroxide using collidine as a buffer substance. These and other results strongly suggested that the histochemical effect of collidine was due to depletion of monoamines stored in the vesicles core. To examine this hypothesis we studied in this work the effect of collidine on tissues that have taken up tritiated noradrenaline. It was found that tritium was released very rapidly to the incubation medium when collidine was applied to fresh tissues. This effect was not observed with other commonly used buffers such as cacodylate or phosphate. It was also found that tritium release also occurred, although to a lesser extent, when tissues were fixed in glutaraldehyde or osmium tetroxide using collidine as a buffer, and this release was not significant when collidine was applied to previously fixed tissues. Paper chromatographic analysis showed that the radioactive compound(s) extracted from tissues by collidine corresponded to noradrenaline and/or closely related compounds. An abstract of this work was sent to the 17th Annual Meeting of the Society for Neuroscience, New Orleans, Nov 16-21, 1987. Tomsig J.L. and Pellegrino de Iraldi A. Abstract 369-11.

Animals↗
PubMed1993

Botulinum B toxin as an alternative to botulinum A toxin: a histologic study.

Histochemical effects of botulinum B toxin were studied on fibers from longissimus dorsi muscle in Albino rabbits and compared to effects produced by botulinum A toxin. Acetylcholinesterase staining, muscle fiber size analysis, and ATPase staining indicated botulinum B toxin produced a denervation gradient and field similar to that produced by botulinum A toxin. At 5 weeks postinjection with botulinum B toxin, analysis showed muscle fiber size variability, and diffuse acetylcholinesterase fiber staining comparable to botulinum A toxin at the injection site. Muscle sections taken at 4.0 cm for analysis showed statistically significant decreased fiber size variability and contraction of acetylcholinesterase staining pattern for both immunotypes. In addition, the denervation reflected by histochemical staining and fiber size analysis appeared transient and lasted for approximately 3 months for both immunotypes. These findings suggest botulinum B toxin produces pharmacologic effects on innervation of striated muscle similar to botulinum A toxin. Because immunologic tolerance has been demonstrated after therapeutic botulinum A toxin injections, further clinical studies need to be conducted with other immunotypes of toxin with no cross-reactivity to type A.

Acetylcholinesterase↗
PubMed1989

Visualizing mRNA expression in plant protoplasts: factors influencing efficient mRNA uptake and translation.

In this paper we demonstrate that RNA sequences present upstream and downstream of a reporter gene coding region play an important role in determining the amount of protein produced from an mRNA. A translational enhancer, omega, derived from tobacco mosaic virus, when present at the 5'-end of beta-glucuronidase mRNA increased the efficiency of translation 16-fold to 18-fold in electroporated tobacco or carrot protoplasts, and threefold to 11-fold in maize or rice protoplasts. The presence of omega did not alter the half-life of the mRNA in vivo. We also demonstrate for the first time that a minimum polyadenylated tail length of 25 adenylate residues is sufficient to substantially increase the expression and half-life of the reporter mRNA in plants. When in vitro-produced mRNAs were synthesized such that extra sequence was added to the 3'-end of the poly(A) tail, however, the final level of expression was decreased up to 80%. Omega, the translational enhancer, and a poly(A) tail function independently of each other; their combined effect on translation, when both are present in an mRNA, is the multiplication of their individual effects. Histochemical analysis for the presence of beta-glucuronidase in tobacco established that virtually all viable cells receive mRNA during electroporation. Video image analysis of tobacco protoplasts electroporated with luciferase mRNA demonstrated that there is a wide range in the level of expression of this marker. Carrier RNA, when present during electroporation, had only a modest effect on increasing mRNA uptake. Reporter mRNA expression in electroporated protoplasts was directly proportional to the input mRNA up to at least 30 micrograms/ml.

Base Sequence↗
PubMed1998

Participation of intracellular cysteine proteinases, in particular cathepsin B, in degradation of collagen in periosteal tissue explants.

The involvement of cysteine proteinases in the degradation of soft connective tissue collagen was studied in cultured periosteal explants. Using cysteine proteinase inhibitors that were active intracellularly or extracellularly (Ep453 and Ep475, respectively), it was shown that over-all collagen degradation, as measured by the release of hydroxyproline, decreased significantly on inhibition of the intracellular pool of cysteine proteinases by Ep453. This inhibitor also induced an accumulation of intracellular fibrillar collagen in fibroblasts, indicating a decreased degradation of phagocytosed collagen. The extracellular inhibitor, Ep475, had minor or no effects. Histochemical analysis using a substrate for the cysteine proteinases cathepsins B and L revealed a high level of enzyme activity, which was completely blocked in explants preincubated with a selective intracellular inhibitor of cathepsin B, Ca074-Me. Moreover, the cathepsin B inhibitor strongly affected collagen degradation, decreasing the release of hydroxyproline and increasing the accumulation of phagocytosed collagen. These effects were comparable or slightly stronger than those found with the general intracellular inhibitor (Ep453). Taken together, these data strongly suggest that intracellular cysteine proteinases, in particular cathepsin B, play an important role in the digestion of soft connective tissue collagen.

Animals↗
PubMed2004

Alterations in enzyme histochemical characteristics of the masseter muscle caused by long-term soft diet in growing rabbits.

OBJECTIVE: Recently young people have an increasing tendency to intake an easily chewable diet and spend less time on mastication. The aim of the present study was to investigate the histochemical effects of long-term soft diet on the masseter muscle in growing rabbits. MATERIALS AND METHODS: Twelve young male Japanese white rabbits were divided into two groups (n = 6 each) at weaning (1 month after birth) and fed a solid diet (control group) or a powder diet (soft-diet group). The duration of the experimental period was 6 months. Masseter fibers from the superficial and the deep portions were histochemically defined as type 1, 2A, 2B, or 2C fibers. RESULTS: As compared with that of the control, the deep masseter of the soft-diet group showed a significantly lower ratio of type 1 fiber cross-sectional area to total area (6.3 and 10.1% for the soft-diet and control group, respectively), significantly more type 2A fibers (74.0%vs 50.3%) and significantly fewer type 2B fibers (4.3%vs 12.5%). However, fiber size did not differ between the two groups. NADH-tetrazolium-reductase (NADH-TR) of the masseter was less reactive in the soft-diet group, reflecting a lower oxidative capacity. CONCLUSIONS: These findings indicate that the alteration of the functional activities contributed to selective disuse influences on the type 1 and type 2B fibers, and a resultant increase in type 2A fibers. This study suggests that long-term alteration of jaw function induced by a soft diet can lead to adaptations of the masseter muscle.

Adaptation, Physiological↗
PubMed1987

Reactivation of inhibited bone acid phosphatase and its significance in bone histomorphometry.

Despite biochemical demonstration of acid phosphatase (AcP) activation or reactivation in bone, few attempts have been made to show similar effects histochemically. Bones from growing rats, when fixed in 4% buffered formaldehyde at room temperature and demineralized in 5% formic acid, exhibited expected inactivation of AcP. The inhibited AcP, however, was reactivated by pre-incubation of sections for 1 hr at 37 degrees C in the following buffers: 0.2 M Tris, 0.2 M glycine, 0.2 M NaHCO3, or 0.1 M borax, as well as in alkaline water, but not in 0.2 M Na2HPO4 (all at pH 9). The reactivation was (a) site-specific (e.g., osteoclasts, osteoblasts, osteocytes, and cement lines), (b) temperature- and pH-dependent, (c) unaffected by OH- or SH--binding agents or by an alkaline phosphatase inhibitor, and (d) inhibited completely by 10 mM Na2HPO4. The reactivation process, much simplified and/or more effective than with the methods previously reported, was observed in all 83 human biopsy bones embedded in methyl methacrylate and in human bones stored in cold buffered formaldehyde for 7 months. This study demonstrates a unique method for reactivating and thus localizing the inhibited AcP in bones, and suggests possible applications in bone histomorphometry.

Acid Phosphatase↗
PubMed1998

Melatonin increases striatal dopaminergic function in 6-OHDA-lesioned rats.

The purpose of this study was to assess the in vivo effects of melatonin, as an antioxidant, on striatal dopaminergic function in rats with a unilateral 6-hydroxydopamine (6-OHDA) lesion of the striatum. Compared with sham-operated controls and expressed as a ratio relative to the contralateral side, there was an increase in the lipid peroxidation product malondialdehyde (MDA, 142%) and a significant reduction in tyrosine hydroxylase (TH) enzyme activity (28%) and dopamine (DA, 32%) and its metabolite dihydroxyphenylacetic acid (DOPAC, 50%) 2 weeks after 6-OHDA injection. Melatonin treatment almost completely restored MDA levels to normal, suggesting the in vivo action of melatonin as an antioxidant. In parallel, partial, but statistically significant recovery of striatal dopaminergic function, including TH enzyme activity and DA levels, also occurred following melatonin treatment. Taken together with our previous reports showing behavioral and histochemical effects of melatonin on the nigrostriatal dopaminergic system, the present results strongly support the hypothesis that melatonin, as an antioxidant, may have beneficial effects on therapeutic approaches for the treatment of oxidative stress-induced neurodegenerative disease such as Parkinson's disease (PD).

3,4-Dihydroxyphenylacetic Acid↗
PubMed1990

Spatial differences of endogenous lectin expression within the cellular organization of the human heart: a glycohistochemical, immunohistochemical, and glycobiochemical study.

Protein-carbohydrate recognition may be involved in an array of molecular interactions on the cellular and subcellular levels. To gain insight into the role of proteins in this type of interaction, surgically removed specimens of human endomyocardial tissue were processed for histochemical and biochemical analysis. The inherent capacity of these sections to bind individual sugar moieties, which are constituents of the carbohydrate part of cellular glycoconjugates, was assessed using a panel of biotinylated neoglycoproteins according to a standardized procedure. Together with appropriate controls, it primarily allowed localization of endogenous lectins. Differences in lectin expression were observed between layers of endocardial tissue, myocardial cell constituents, connective-tissue elements, and vascular structures. The endocardium proved to be positive with beta-galactoside-bearing probes; with neoglycoproteins carrying beta-xylosides, alpha-fucosides, and galactose-6-phosphate moieties; and with probes containing a carboxyl group within the carbohydrate structure, namely sialic acid and glucuronic acid. In contrast, only fucose-and maltose-specific receptors were apparent in the elastic layers of the endocardium. Aside from ascertaining the specificity of the protein-carbohydrate interaction by controls, i.e., lack of binding of the probe in the presence of the unlabelled neoglycoprotein and lack of binding of the labelled sugar-free carrier protein, respective sugar receptors were isolated from heart extracts by using histochemically effective carbohydrates as immobilized affinity ligand. Moreover, affinity chromatography using immobilized lactose as affinity ligand as well as the use of polyclonal antibodies against the predominant beta-galactoside-specific lectin of heart demonstrated that the lactose-specific neoglycoprotein binding was due to this lectin. Remarkably, the labelled endogenous lectin, preferred to plant lectins for detecting ligands of the endogenous lectin, localized ligands in tissue parts where the lectin itself was detected glycohistochemically as well as immunohistologically. This demonstration of receptor-ligand presence in the same system is a further step toward functional assignment of the recorded protein-carbohydrate interaction. Overall, the observed patterns of lectin expression may serve as a guideline to elucidate the precise physiological relevance of lectins and to analyze pathological conditions comparatively.

Endocardium↗
PubMed1982

Endothelium and "silver lines". An electron microscopic study.

The significance of endothelial "silver lines" was studied by TEM in rat aortas after perfusion with glutaraldehyde followed by silver nitrate. Standard TEM technique proved unsatisfactory (coarse silver granules, imprecise localization, artefacts). Exposure of the silver-treated aortas to photographic fixer markedly improved the image of the deposits leaving fine, stable, uniform "residual granules" about 100 A in diameter. Most of these granules were localized along the intercellular junctions; they also tended to pool in the basement membrane beneath each junction. This image suggests that the Ag+ ions pass through the junction, and react with its contents as well as with the basement membrane beyond it. A scheme is proposed to explain the reaction of Ag+ ions with anions and negatively charged radicals within the junction. It is concluded that the "silver lines" represent not only a histochemical effect, but also the visualization of a transendothelial electrolyte pathway.

Animals↗
PubMed1984

6-Hydroxydopamine produces lesions of serotonin-containing Retzius cells in the leech nervous system.

The neurotoxin 6-hydroxydopamine (6-OHDA) is reported to selectively ablate neurons which contain catecholamines. Leeches were injected with 6-OHDA over a 4-month period in an attempt to destroy their identified dopamine neurons. Ganglionic Retzius cells (RZ), which contain serotonin, were the first to be rendered brown, misshapen, and non-fluorescent. Continued injections of 6-OHDA had similar, but less substantial, morphological and histochemical effects on the dopamine-containing neurosomata within the anterior roots. Toxin treatment reduced RZ serotonin by 75-90%, but these depleted cells retained normal electrophysiological properties. Serotonin and dopamine within ganglia, as well as dopamine within anterior roots, were reduced significantly by 6-OHDA.

Animals↗