Tetrazolium Reductase Research Articles

Adrenal changes in Niemann-Pick disease: Differences between sphingomyelinase deficiency and type C

Structural, chemical, and histochemical analyses of adrenal tissue performed in 8 cases of Niemann-Pick disease (NPD) revealed stark differences of storage between spingomyelinase (SMase) deficiency (6 cases) and type C (2 cases). In all the full-blown cases of the SMase deficiency group, pronounced sphingomyelin (SM) storage was found in all the zones of the cortical epithelium with slightly increasing centripetal gradient. The storage resulted in the reduction or even disappearance of lipofuscinogenesis in the reticular zone, in the reduction of the physiological fat content, in the generalized foamy transformation of the epithelium, and in moderate organomegaly. The storage was expressed in both A and B types and was roughly proportional to the storage in other viscera. The stromal storage was confined to the vascular endothelium, and in particular, to the macrophages. One of the cases showed the presence of typical spirolactone bodies unmodified in fine structure by the lysosomal storage. Their most conspicuous enzymatic activity was that of non-specific esterase and NADH tetrazolium reductase. The adrenals in type C were macroscopically and histologically normal except for a variable population of stromal foam cells. Chemically, there was slight increase in all phospholipids with borderline or moderate percentual increase of SM. There was also slight increase in some of the lower neutral glycosphingolipids. Electron microscopy dislosed rudimentar storage in lower cortical layer epithelium which by its fine structure and according to results of lipid histochemistry was qualitatively different from that in SMase deficiency. The stromal storage was expressed mainly in macrophages in which there was histochemically detectable amount of SM. There was no storage detectable in medullary cells in neither group of NPD complex. The results point not only to striking quantitative differences in storage intensity between the 2 basic groups of NPD showing the cortical epithelium in type C as being remarkably resistant to the metabolic disorder, but also to difference in quality of the storage very much like that found in other tissues, too.

Muscle Histochemistry of the Planarian Dugesia tigrina (Turbellaria: Tricladida): Implications in the Evolution of Muscle

Muscle Histochemistry of the Planarian Dugesia tigrina (Turbellaria: Tricladida): Implications in the Evolution of Muscle

Inhibition of the O 2−-generating NADPH oxidase of guinea-pig polymorphonuclear leukocytes by rabbit antibody to homologous liver NADPH-cytochrome c ( P-450) reductase

Rabbit antibody highly specific for guinea-pig liver NADPH-cytochrome c ( P-450) reductase was found to inhibit dose-dependently the O 2 −-generating activity of the membrane fraction isolated from phorbol-myristate acetate-stimulated, homologous polymorphonuclear leukocytes. In addition, the antibody also could inhibit the NADPH-cytochrome c (Nitroblue tetrazolium) reductase from the membrane fractions and phagosomes of leukocytes by polyacrylamide gel electrophoresis or gel filtration on a Sephacryl S-300 column in the presence of 0.2% Triton X-100. These results demonstrate that the NADPH-cytochrome c reductase in the membrane fractions of leukocytes is antigenically cross-reactive with homologous liver NADPH-cytochrome c reductase, and also suggest that the enzyme of leukocytes participates in the respiratory burst.

Histoenzymological differences in the envelope of white and stages of rapidly growing yellow follicles in the ovary of crow (Corvus splendens)

Histoenzymological differences in the localization and intensities of succinate and lactate dehydrogenases, tetrazolium reductases and steroid dehydrogenases have been studied in the follicular envelope of white and stages of yellow follicles of the crow ovary. The activities of these enzymes in the thecal gland cells show a progressive increase during follicular growth. The granulosa layer of white follicle is devoid of Δ5-3β-hydroxysteroid dehydrogenase which appear and show a gradual increase in the rapidly growing yellow follicles along with other enzymes.

Bone marrow aplasia after high voltage electrical injury

This report describes a patient with severe high voltage electrical injury in whom bone marrow aplasia developed on post-burn day 3. In addition to bone marrow aplasia alterations suggesting disintegration of leucocytes and reduced nitro-blue tetrazolium reductase activity pointing to functional disorders have been observed. Beyond the effect of the extensive burns to depress the bone marrow the possibility of an additional similar action of the high voltage electrical injury is raised.

Histochemical characteristics of the tensor tympani muscle in relation to the medial gastrocnemius muscle of the cat.

Serial 10-micron cryostat cross-sections of the tensor tympani muscle and of the medial gastrocnemius muscle from adult domestic cats were incubated for myofibrillar ATPase, NADH tetrazolium reductase (NADH-TR), succinic dehydrogenase (SDH), malate dehydrogenase (MDH) and menadione-linked alpha-glycerophosphate dehydrogenase (alpha GPDH). The optical density of individual tensor tympani and gastrocnemius muscle fibres after different incubation procedures was measured photometrically. The absorbance values of the tensor tympani fibres were related to the values of the type I, type IIA and type IIB fibres of the gastrocnemius muscle. Only two different types of fibre could be demonstrated in the tensor tympani, one type resembling the type I and another resembling the type IIA of the gastrocnemius muscle. The findings are discussed in relation to other, recent immunohistochemical studies on cat tensor tympani muscle fibres.

D-T diaphorase and NAD(P)H-NBT reductase activation

The effects of 3-methylcholanthrene, benz(a)pyrene and 7,12-dimethylbenz(a)anthracene on NAD(P)H dehydrogenation were studied in the mesenteric lymph nodes of the rat. The biochemical studies were performed in three different fractions, one fraction sedimented at 10,000 g, the other at 100,000 g and in the supernatant of the cells. The dehydrogenation of NAD(P)H by the soluble enzyme D-T diaphorase was increased several fold by the three chemicals, while the particulate enzymes which dehydrogenate NAD(P)H were not affected by the actions of these carcinogens. The enzymatic changes were dose dependent and were inhibited by dicumarol. Histochemically, the administration of the carcinogens markedly increased the NAD(P)H-nitroblue tetrazolium reductase activity in the cortical and in paracortical lymphocytes and in histiocytes. The histochemical reaction of the cells in the germinal centers was negative, even after the administration of the carcinogens. A correlation between the biochemical and histochemical finding are attempted in this communication.

Capillarity and fiber types in the cremaster muscle of rat and hamster.

We determined muscle fiber type and capillarity in cremaster muscle samples from rats and hamsters of different ages. Histochemical estimation of oxidative capacity was made from the activity of either nicotinamide dinucleotide tetrazolium reductase (NADH-TR) or succinic dehydrogenase (SDH), and fibers were termed fast or slow from myofibrillar ATPase activity. Fibers were classified as type I (low ATPase, high NADH-TR/SDH), type IIa (high ATPase, high SDH/NADH-TR), type IIb (high ATPase, low SDH/NADH-TR), or type IIc (no acid reversal of ATPase, high NADH-TR). Type IIb fibers accounted for 60-80% of the muscle area in both species at all ages. The principal change with maturation was muscle fiber hypertrophy. Mean cross-sectional fiber area increased from 488 +/- 70 (SE) and 453 +/- 19 micron2 in young hamsters and rats, respectively, to 1,255 +/- 99 and 1,540 +/- 101 micron2 in adults. Capillary density (no. of capillaries/mm2 tissue) paralleled fiber hypertrophy; it decreased significantly with maturation from 684 +/- 60 (SE) to 228 +/- 26/mm2 in hamsters and from 341 +/- 15 to 213 +/- 15/mm2 in rats. In vitro estimates of capillary density are compared with previously obtained in vivo data (31), and sources of error are identified. We conclude that reported differences in microvascular function in the cremaster muscle in vivo during maturation or between species cannot be ascribed to changes in muscle composition.

Competitive ability, effectiveness, and succinate – triphenyl tetrazolium chloride reductase activity of mutant strains of chick-pea (Cicer arietinum L.) Rhizobium sp.

Chick-pea Rhizobium strain RG 4 was treated with 200 μg/mL of N-methyl-N-nitro-N′-nitrosoguanidine and 14 mutant strains isolated. Two mutant strains were found to be nonnodulating in the absence of certain amino acids. On the basis of nodulation, two mutant strains (M 38 and M 46) were found to be equally competitive against each other. Mutant strains M 38 and M 46 when applied together produced the maximum nodulation, nitrogenase activity, phenanthroline-reactive iron, and least tetrazolium reductase activity as compared to other pairwise combinations of mutant strains. It was also observed that the correlation between tetrazolium reductase and nitrogenase activity was highly negative.

Histochemical distribution of tetrazolium reductases, dehydrogenases and lipids in the follicular wall of normal and atretic follicles in the ovary of the Indian gerbilTatera indica (Muridae: Rodentia)

The comparison of the histochemical features of the follicular wall in normal and atretic follicles of corresponding stages in the ovaries of Indian gerbilTatera indica revealed conspicuous changes in amount of lipids and activities of NADH- and NADPH tetrazolium reductases, δ5-3β-hydroxysteroid dehydrogenase and 17α-hydroxysteroid dehydrogenase and succinate- and lactate dehydrogenases in the granulosa layers during follicular growth and atresia. The thecal layers showed a progressive increase in their amounts during follicular growth and these enzymes are not affected significantly by atresia. Atresia resulted in increased amounts of lipids in the granulosa cells and decreased activities of succinate- and lactate dehydrogenases and NADH- and NADPH tetrazolium reductases during latter stages of degeneration.

Isozyme patterns and morphogenesis in higher basidiomycetes

Earlier studies on sexual morphogenesis in higher basidiomycetes had indicated that there were major changes in the electrophoretic mobility and in the number of isozymes of several metabolic enzymes during the transition from homokaryon to dikaryon. Work with the malic dehydrogenase isozymes of Coprinus congregatus and Schizophyllum commune has not uncovered any isozyme shifts comparable to those reported earlier. The difference in the results of this work from the earlier ones is attributed, in part, to changes in technique involving exogenous electron carriers. Evidence is presented involving endogenous tetrazolium reductases and/or pyridine nucleotide dehydrogenases as the causal agents of artifactual bands. In addition, it is shown that the synthesis of the same proteins is regulated by nuclei in either the homokaryon or the dikaryon state and that in the dikaryon state there can be normal subunit aggregation of monomers to produce hybrid proteins. This evidence is discussed in relation to recent works that refute and confirm the earlier studies.

A fixative for use in muscle histochemistry.

A fixative solution that preserves the activity of some relevant enzymes in muscle histochemistry is described. Portions of human muscle biopsy specimens and selected murine muscles were fresh frozen or placed in the fixative at room temperature for up to 1 month before freezing. Cryostat sections of fresh frozen and fixed frozen tissue were assayed for nicotinamide adenine dinucleotide phosphate (NADH)-tetrazolium reductase (NADH), several adenosine triphosphatases (ATPases), myoadenylate deaminase (MD), and phosphorylase. NADH, ATPase, and MD activity were preserved following fixation but phosphorylase was not preserved. Murine spleen and kidney were similarly tested for acid phosphatase (acid phos), alkaline phosphatase (alk phos), and nonspecific esterase (NSE). Alk phos activity was preserved but acid phos and NSE activity were significantly reduced following fixation. This fixative is useful in some circumstances for processing or shipping human muscle biopsy specimens and experimental tissues.

The influence of polycyclic hydrocarbons on the activity of NAD(P)H-dehydrogenating enzymes in rat thymus. A biochemical and histochemical study.

The action of benzo(a)pyrene, 3-methylcholanthrene and 7,12-dimethylbenzo(a)anthracene in the activity of the rat thymus D-T diaphorase (EC 1.6.99.2) and the NAD(P)H cytochrome C (EC 1.6.99.3) reductases of particulate fractions were studied in intact and adrenalectomized animals. These polycyclic hydrocarbons increased severalfold the activity of the D-T diaphorase in intact and adrenalectomized animals. The activities of the particulate enzymes were not affected by the carcinogens. Dicumarol suppresses the inducing effects of benzo(a)pyrene and adrenalectomy does not influence the inducing effects of benzo(a)pyrene and 3-methylcholanthrene. The histological distribution of the enzyme NAD(P)H-nitroblue tetrazolium reductase was studied and a marked increase in its activity in lymphocytes, macrophages and epithelial cells was found after the administration of the carcinogens.

New enzymatic findings in infantile neuroaxonal dystrophy.

Two siblings with infantile neuroaxonal dystrophy (INAD) exhibited highly increased activity of nonspecific esterase (resistant to physostigmine and sensitive to organophosphates) topically correlating with the morphological manifestations of the disease (spheroids), confined in those two particular cases to the central nervous system (CNS). This is demonstrated for the first time. NADH tetrazolium reductase was another enzyme noted with increased activity and similar topography. The lesions were devoid of any marked activity of typical mitochondrial (succinate and alpha-glycerolphosphate) dehydrogenases and lysosomal (acid phosphatase and beta-glucuronidase) enzymes. The diagnostic significance of monitoring non-specific esterase and NADH tetrazolium reductase activities in the diagnosis of INAD is discussed in particular.

Macroscopic enzyme histochemistry in myocardial infarction: artefactual nature of the creatine phosphokinase reaction.

The histochemical creatine phosphokinase (CPK) tetrazolium test has been evaluated to detect recent human myocardial infarction in gross slices of the heart at necropsy. The demonstration of the lesion with this method has been assumed to result from local loss of CPK from the damaged myocardium. However, the present study indicates that the mechanism involved depends on localising NADPH tetrazolium reductase and not CPK. Phenazine methosulphate (PMS), when added to the incubating medium as an electron-acceptor to circumvent the tetrazolium-reductase (diaphorase) system, resulted in generalised false staining of the heart slice.

Morphometric studies on tenuissimus muscle spindles in the cat.

Muscle spindles were studied histochemically in serial transverse sections of 42 cat tenuissimus muscle specimens. Staining for myofibrillar adenosine triphosphatase was employed to identify nuclear bag 1, nuclear bag 2, and nuclear chain intrafusal muscle fibers. The nuclear chain fibers were further subdivided into three categories according to their polar length and the intensity of their staining for nicotinamide adenine dinucleotide tetrazolium reductase. A total of 430 spindle poles were surveyed. The mean spindle content of bag 1, bag 2, and chain fibers was established. The mean polar length of intrafusal fibers as well as that of the intracapsular and extracapsular spindle regions was determined. A cholinesterase (ChE) staining technique was used to demonstrate the termination sites of motor axons along intrafusal fibers. Two types of circumscribed ChE deposits. The "rim" and the "plate," occurred on the fibers. The nuclear chain fibers usually carried both the ChE rims and plates, while most nuclear fibers displayed only the plates. The ChE plates were assessed in term of their appearance, staining intensity, length, and location along the fibers. The mean number of ChE plates found along the fibers was established for each of the various intrafusal fiber types. These histochemical observations are discussed with regard to the current concepts of cat spindle morphology and motor innervation. The results suggest a degree of predictability in the spindle fiber content and in the distribution of motor nerve terminals along intrafusal muscle fibers, at least in the tenuissimus muscle.

Electrical stimulation of denervated muscle prevents decreases in oxidative enzymes.

The influence of muscular contraction on the oxidative enzymes and the diameters of muscle fibers was investigated. Soleus muscles of guinea pigs were denervated for four weeks. The denervated fibers showed a reduction in the intensity of staining for beta-hydroxybutyrate dehydrogenase, cytochrome oxidase, succinate dehydrogenase, and NADH-dependent tetrazolium reductase. Denervation also resulted in a decrease in fiber diameter. Denervated soleus muscles were electrically stimulated to contract over a four-week period at a frequency normally received by slow contracting muscles. Electrical stimulation caused the stain intensity of histochemical reactions for oxidative enzymes to appear to be normal or greater than normal in 90% of the denervated fibers. Stimulation also caused 69% of the denervated fibers to be of normal or greater than normal size. The results demonstrate that contraction of denervated muscle by electrical stimulation prevents the loss of oxidative enzymes and the atrophy associated with denervation.

Examination of chronically de-efferented cat muscle spindles for cholinesterase activity.

Cat tenuissmus muscles were deprived of motor nerve supply for three months by sectioning of the appropriate ventral spinal roots. Muscle spindles were located in the chronically de-efferented muscles and examined histochemically in serial transverse sections. Staining for nicotinamide adenine dinucleotide tetrazolium reductase showed that the spindle sensory innervation was preserved. The de-efferented intrafusal muscle fibers retained their differential staining with the reaction for myosin adenosine triphosphatase. However, all cholinesterase-active areas that are normally found along nuclear bag and nuclear chain intrafusal fibers demonstrated loss of the enzyme activity in the chronically de-efferented spindles. It is concluded that all histochemically demonstrable cholinesterase activity within the cat muscle spindle is dependent upon the continuous presence of motor innervation.

Distribution of fiber types in locomotory muscles of dogs.

The distribution of Type I and Type II fibers, as determined from histochemical estimation of myofibrillar ATPase activity, was studied within and among the locomotory muscles of the forelimb, trunk, and hindlimb of three mongrel dogs. All Type II fibers had high oxidative capacities as estimated from the histochemical assay for reduced nicotinamide adenine dinucleotide tetrazolium reductase, so they were not further divided into subpopulations. Furthermore, Type I and Type II fibers had similar oxidative potentials as indicated by both histochemistry and biochemistry. Type I fiber populations ranged between 14% and 100% in the muscles sampled. The highest percentages of Type I fibers were found in deep muscles of physiological extensor groups in the arm and thigh that serve to resist gravity (antigravity muscles) when the dog is in the quadrupedal standing position. More superficial muscles in these same groups had fewer Type I fibers. The patterns of Type I fiber distribution among muscles in the antigravity groups of the forearm and leg were the opposite of those in the arm and thigh, with the more superficial muscles of the distal limb segments having more Type I fibers than the deeper muscles. In all limb segments, muscle groups that do not serve to resist gravity did not show as much intermuscular variation. Type I fiber populations in these muscles did not exceed 50%. A stratification of fiber types also existed within muscles, both in extensor and flexor groups, with the deeper portions of the muscles having more Type I fibers than the more superficial portions.

Toxicity of diminazene aceturate (Berenil) to camels

Ten mg per kg of diminazene aceturate (Berenil) was highly toxic to camels. The main signs of berenil poisoning were hyperaesthesia, salivation, intermittent convulsions, frequent urination and defaecation, itching and sweating. The damage to the liver and kidney was accompanied by increases in the activity of ASAT and in the concentration of ammonia and by decreases in the concentration of calcium and magnesium in serum. The activity of adenosine triphosphatase, 5-nucleotidase, succinic tetrazolium reductase and glucose-6-phosphatase was reduced in the hepatocytes and the cells of the renal tubules.