Nicotinamide Adenine Dinucleotide Diaphorase Activity Research Articles

29 Attitude and behavior of veterinarians surrounding the use of complementary and alternative veterinary medicine (CAVM) in the treatment of equine musculoskeletal pain

The skeletal muscle has several types of fibers that differ in structure, metabolism, and function. Fiber types determine the ability of horses to perform in various sports or work activities. Gluteus medius muscles of Colombian creole horses (CCHs) were sampled to determine their fiber types. Samples were processed using myosin adenosine triphosphatase (mATPase) and immunohistochemistry for fiber typing, myofibril electrophoresis for myosin heavy chain composition, and nicotinamide adenine dinucleotide diaphorase activity for oxidative capacity. The following fiber proportions were observed by immunohistochemistry (n = 7): 22.5 ± 3.7% type I, 33.6 ± 2.3% type IIA, 43.6 ± 4.4% type IIX, and 0.3 ± 0.2% hybrid I/IIA fibers. The results obtained by mATPase were comparable. Regarding the oxidative capacity, 20.1 ± 4.0% had a high oxidative reaction, 32.7 ± 6.1% an intermediate reaction, and 47.2 ± 7.4% a low oxidative reaction. Of these, the high and low capacity correlated with type I and IIX fibers, respectively. Myosin heavy chain composition experiments (n = 7) showed 22.2 ± 1.6% of type I, 30.4 ± 2.7% of type IIA, and 47.4 ± 2.3% of type IIX. These values are close to those published for Andalusian horses but differ from some reported for other breeds. Considering the high percentage of type IIX fibers and low to intermediate oxidative capacity, we conclude that the CCH is adapted to perform power exercise and moderate- to high-intensity anaerobic work with short to average duration. Therefore, training should be aimed at increasing aerobic capacity and endurance if these animals are required to perform long-length exercise.

Muscle Fiber Types of Gluteus Medius in the Colombian Creole Horse

The skeletal muscle has several types of fibers that differ in structure, metabolism, and function. Fiber types determine the ability of horses to perform in various sports or work activities. Gluteus medius muscles of Colombian creole horses (CCHs) were sampled to determine their fiber types. Samples were processed using myosin adenosine triphosphatase (mATPase) and immunohistochemistry for fiber typing, myofibril electrophoresis for myosin heavy chain composition, and nicotinamide adenine dinucleotide diaphorase activity for oxidative capacity. The following fiber proportions were observed by immunohistochemistry (n = 7): 22.5 ± 3.7% type I, 33.6 ± 2.3% type IIA, 43.6 ± 4.4% type IIX, and 0.3 ± 0.2% hybrid I/IIA fibers. The results obtained by mATPase were comparable. Regarding the oxidative capacity, 20.1 ± 4.0% had a high oxidative reaction, 32.7 ± 6.1% an intermediate reaction, and 47.2 ± 7.4% a low oxidative reaction. Of these, the high and low capacity correlated with type I and IIX fibers, respectively. Myosin heavy chain composition experiments (n = 7) showed 22.2 ± 1.6% of type I, 30.4 ± 2.7% of type IIA, and 47.4 ± 2.3% of type IIX. These values are close to those published for Andalusian horses but differ from some reported for other breeds. Considering the high percentage of type IIX fibers and low to intermediate oxidative capacity, we conclude that the CCH is adapted to perform power exercise and moderate- to high-intensity anaerobic work with short to average duration. Therefore, training should be aimed at increasing aerobic capacity and endurance if these animals are required to perform long-length exercise.

Selective Prostate Cancer Thermal Ablation With Laser Activated Gold Nanoshells

Laser activated gold nanoshell thermal ablation represents a new, minimally invasive technology that offers benign tissue sparing thermal ablation of malignant tumors. We evaluated the efficacy of this technology for eradicating prostate cancer in a subcutaneous tumor model. The 110 nm gold nanoshells with a 10 nm gold shell are designed to act as intense near infrared absorbers. PC-3 cells were injected on the dorsum of nude mice in 3 groups, including 1-gold nanoshell plus near infrared laser, 2-saline alone and 3-near infrared laser alone. Animals received 7.0 ml/gm body weight (low dose) or 8.5 ml/gm body weight (high dose) nanoshells via tail vein injection. Control animals received saline. A 810 nm near infrared laser with a 200 mu laser fiber and an energy setting of 4 W/cm(2) was aimed at the tumor bed for 3 minutes. Tumors were measured at days 0, 7, 14 and 21. Tissue temperature was monitored during laser activation. Tumors were harvested at day 21 and stained with hematoxylin and eosin, and for nicotinamide adenine dinucleotide diaphorase activity. We observed 93% tumor necrosis and regression in the high dose treated group. Nicotinamide adenine dinucleotide staining corroborated this finding. The ablation zone was sharply limited to the laser spot size. There was no difference in the size or tumor histology in control groups, indicating a benign course for near infrared laser treatment alone. Temperatures up to 65.4C were attained in the treated group. Laser activated gold nanoshell ablation is an effective and selective technique for prostate cancer ablation in an ectopic murine tumor model.

Feasibility of New Heating Method of Hepatic Parenchyma Using a Sintered MgFe 2O 4 Needle Under an Alternating Magnetic Field

Magnetic metal particles such as magnesium ferrite (MgFe2O4) induce heat energy under an alternating magnetic field that was produced by electric current. We have developed a new heating device using a sintered MgFe2O4 needle under an alternating magnetic field. This device can repeatedly heat target tissue at lower temperatures than that for radiofrequency ablation therapy. This study aims to assess whether the new heating device has the ability to heat rat liver tissue. A small needle made from MgFe2O4 particles was prepared by sintering at 1100 degrees C and inserted into rat liver tissue. The rat liver was then heated under an alternating magnetic field, 4 kA/m, for 30 min. We measured the temperature of rat tissue during the heat treatment, and sequentially evaluated histological changes and hepatocyte cellular activity after heat stimulus by using nicotinamide adenine dinucleotide diaphorase staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. The mean temperature of the liver tissue during heating was 60.7 +/- 1.1 degrees C. Immediately after heating, nuclei of the hepatocytes were hyper-chromatin, with hepatocytes negative for nicotinamide adenine dinucleotide diaphorase activity in the heat-injury area. The injury area spread progressively until 3 d after heating, when the area was surrounded by fibroblasts, with hepatocytes positive for terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. This is the first time that a ferromagnetic metal heating device under an alternating magnetic field has achieved a temperature beyond 60 degrees C and led hepatocytes to complete cell death. This device would be of future use as a local heat-treatment for human liver cancer.

Nicotinamide Adenine Dinucleotide Staining Immediately Following Radio Frequency Ablation of Renal Tumors—Is a Positive Stain Synonymous With Ablative Failure?

Nicotinamide adenine dinucleotide diaphorase staining is arguably the standard for assessing tissue viability following radio frequency ablation, yet the accuracy of this test is questionable. Thus, it is imperative to examine the reliability of nicotinamide adenine dinucleotide to predict ablation success. To evaluate the observation that immediate nicotinamide adenine dinucleotide staining may not be clinically reliable, we compared results from immediate post-ablation biopsies of renal tumors to radiological and clinical followup. Laparoscopic radio frequency ablation was performed in 9 patients and 10 tumors using a temperature modulated radio frequency system. Cold cup biopsies were taken immediately following ablation, and processed for nicotinamide adenine dinucleotide and hematoxylin and eosin staining. Patients were then followed using contrast enhanced computerized tomography at regular intervals. Median tumor size was 2.3 cm. Hematoxylin and eosin stain diagnosed 8 renal cell carcinomas and 2 angiomyolipomas. A quarter of the renal cell carcinomas and both angiomyolipomas stained positive for scattered nicotinamide adenine dinucleotide diaphorase activity immediately after RFA. Mean followup for the nicotinamide adenine dinucleotide positive tumors was 28.5 months (range 24 to 30) and for the nicotinamide adenine dinucleotide negative tumors was 25 months (range 18 to 30). There was no evidence of local tumor recurrence in any patient. Four lesions had nicotinamide adenine dinucleotide diaphorase activity on post-ablation biopsy suggesting retained viable tissue, yet there has been no recurrence during an average 2-year followup. While negative nicotinamide adenine dinucleotide staining is consistent with nonviability, these results suggest that false-positive staining can occur immediately following RFA, making the predictive value of positive nicotinamide adenine dinucleotide diaphorase staining unclear.

Exercise Performance and Skeletal Muscles in Patients With Advanced Chagas Disease

This study was designed to evaluate the peripheral muscle metabolic and structural characteristics in patients with advanced Chagas disease (ChD), and whether they were related with exercise performance. Cross-sectional study. Outpatient cardiomyopathy clinic of a university hospital. We studied 11 stage II patients, 8 stage III patients, and 11 healthy volunteers. All patients underwent exercise testing and peripheral muscle biopsies. The muscles were also studied in control subjects. Muscle biopsy specimens were analyzed for histochemical characteristics. In six patients, the muscle was studied ultrastructurally. The data demonstrate more glycolytic and less oxidative capacity of the peripheral muscle in patients with advanced ChD (increased proportion of type IIb fibers, increased proportion of fibers with low nicotinamide adenine dinucleotide diaphorase activity, high proportion of darkly stained fibers for alpha-glycerophosphate dehydrogenase activity, and lower levels of citrate synthase). Many capillaries in patients with ChD had an abnormal aspect: they were either closed or showed a thicker wall. The ultrastructural study also showed fiber atrophy and abnormal capillaries even in patients with normal functional capacity. Some muscle characteristics (enzyme activity, mean cross-sectional area of the fiber, and capillarity) related with exercise parameters (anaerobic threshold, and peak oxygen pulse). These findings indicate that patients with advanced ChD have decreased oxidative capacity and a shift to anaerobic metabolism in the skeletal muscle. They also suggest that muscular abnormalities are related to oxygen delivery, which is probably reduced in part by the abnormal muscle microvasculature. Those changes could affect oxygen extraction, and therefore exercise tolerance in these patients.

The pretectal complex of the rabbit: distribution of acetylcholinesterase and reduced nicotinamide adenine dinucleotide diaphorase activities.

The chemoarchitecture of the pretectal complex of the rabbit was examined in sections stained by acetylcholinesterase (AChE) and reduced nicotinamide adenine dinucleotide (NADH) diaphorase in the coronal, horizontal and sagittal plane. Twelve different subdivisions can be identified in the rabbit pretectum on the basis of the distribution of both histochemical markers. According to the standard terminology, the pretectal complex of the rabbit consists of: the nucleus of the optic tract; the anterior, posterior, olivary and medial pretectal nuclei; the nucleus of the posterior commissure; the periventricular subcommissural gray; the suprageniculate and internal suprageniculate nuclei, and the dorsal, lateral and medial terminal nuclei of the accessory optic system. The combined use of several sectioning planes and the histochemical mapping of AChE and NADH diaphorase have been of value in resolving the structural limits within transitional regions of the pretectum.

Enzyme histochemical profiles of fish spinal motoneurons after cordotomy and axotomy of motor nerves

Histochemical profiles were made of identified spinal motoneurons from normal adult zebrafish and from animals subjected to cordotomy or unilateral axotomy of the motor nerves. The lesions caused an increase of the myotomal area with oxidative muscle fibers. We studied the question: do changes in the myotomal muscle configuration concur with changes in the enzyme histochemical profiles of innervating motoneurons? Based on the location and size of cell somata, two categories of motoneurons are distinguished: large white (W) motoneurons that innervate the deep fast, glycolytic muscle fibers, and smaller red and intermediate (RI) motoneurons that innervate the superficial slow oxidative and intermediate muscle fibers. In normal animals, glucose-6-phosphate dehydrogenase activity is high in the large W motoneurons and relatively low in the small RI motoneurons. The reverse holds for succinate dehydrogenase activity. W and RI motoneurons show similar nicotinamide adenine dinucleotide diaphorase activity. Short- (2 weeks) and long- (8 weeks) term effects of lesions were studied. The results show that: (1) the 3 types of lesions lead to prolonged changes in the enzyme histochemical profiles of spinal motoneurons. The type of change depends on the type of lesion and on the type of motoneuron; (2) unilateral axotomy of the motor nerves affects the histochemical characteristics of spinal motoneurons and the myotomal muscle fiber type configuration on the ipsi- and contralateral side. The contralateral effects are conceived as adaptations to maintain a left-right symmetry in the motor output.

Properties of immobilized hind-limb muscles of the Galago senegalensis

Morphological, biochemical, and physiological properties of longterm (6 mo) immobilized skeletal muscles of a nonhuman primate were studied in seven Galago senegalensis which were immobilized at the ankle and knee of one hind limb with an external brace. Electromyographic activity of the ipsilateral and contralateral quadriceps and muscles of the calf were assessed after 5 and 6 mo of immobilization. The EMG was markedly reduced in the immobilized muscles compared to the contralateral control when the brace was intact and the animal moving freely. Without exception, extensor muscles atrophied more than flexors. The soleus, a slow-twitch muscle atrophied more than any other muscle of the lower leg but the same was not true of the vastus intermedius the analogous muscle of the thigh. Slow-twitch oxidative fibers (SO) were atrophied more than fast-twitch oxidative glycolytic fibers (FOG), and FOG tended to atrophy more than fast-twitch glycolytic fibers (FG). The immobilized soleus and vastus intermedius had a smaller percentage of SO fibers than their controls, suggesting that they had faster contraction times. With respect to alterations in reduced nicotinamide adenine dinucleotide diaphorase activity, no consistent pattern was observed except for a greater coarseness of staining granules and more homogeneous dispersion of the granules throughout the cross-section of the fibers. No changes were found in phosphorylase, lactate dehydrogenase, or succinate dehydrogenase specific activity or in myoglobin concentration in homogenates of ankle flexors or the vastus lateralis. Myosin ATPase, but not actomyosin ATPase activity was significantly less in the immobilized gastrocnemius-plantaris muscles. No change in contractile properties related to speed were observed in the plantaris. This muscle did exert more twitch and tetanic tension per gram of muscle in the immobilized leg.

Postnatal growth of the extra- and intrafusal fibers in the soleus and medial gastrocnemius muscles of the cat.

AbstractDevelopment of intrafusal and extrafusal fibers in the soleus and medial gastrocnemius muscles in kittens was studied in sections stained by van Gieson or histochemical procedures. Cross‐sectional areas of intrafusal and extrafusal fibers overlapped extensively in the two weeks following birth, with the means for intrafusal fibers in the medial gastrocnemius muscle being larger than those for the extrafusal fibers. A definite increase in extrafusal fiber size was seen at 20 days and in intrafusal fibers at 29 days. In the one mother cat studied, mean areas for soleus and medial gastrocnemius fibers were 12 and 59 times larger than those for a kitten sacrificed the day after birth, while the intrafusal fiber areas were 3.0 and 4.8 times greater. The equatorial region in nuclear bag fibers showed no significant growth in cross‐sectional area, but almost doubled in length. In a newborn kitten, all extrafusal fibers in the soleus and medial gastrocnemius demonstrated a high level of nicotinamide adenine dinucleotide diaphorase activity, but with the adenosine triphosphatase reaction, fibers had either low or high activity. After one month, soleus fibers with high adenosine triphosphatase activity were scarce, whereas in the medial gastrocnemius fibers with both low and high levels of activity were present. The intrafusal fibers at birth were already morphologically and histochemically differentiated into nuclear chain and bag fibers, and two varieties of the latter could be recognized in slides treated for adenosine triphosphatase activity.

Hindlimb muscle fiber populations of five mammals.

The fiber type profiles of hindlimb muscles in the Hartley guinea pig, Sprague-Dawley rat, cat, Galago senegalensis (lesser bushbaby) and Nycticebus coucang (slow loris) were estimated histochemically. Fibers were classified as fast oxidative glycolytic, fast glycolytic or slow oxidative according to their myosin adenosine triphosphatase, α-glycerophosphate dehydrogenase and reduced nicotinamide adenine dinucleotide diaphorase activities. It was found that the soleus and vastus intermedius muscles had the highest proportion of slow oxidative fibers in all five species, demonstrating the constancy of muscle fiber profiles dependent upon anatomical position and functional utilization. The tensor fascia latae and white vastus lateralis of the guinea pig were mostly fast glycolytic, while the red vastus lateralis of the guinea pig consisted of predominantly fast oxidative glycolytic fibers. The majority of muscles investigated in these five mammals were heterogeneous, having a wide range of percentages of the three fiber types.

Types of muscle fibers in the extraocular muscles of birds

Eye muscles of the quail, pigeon, sparrow and canary were examined histologically to see if their fibers are differentiated into two or more types, as has been described for mammals. In each oblique and rectus muscle examined, an area along the orbital border was found to be occupied by fibers of small cross-sectional area, and a more extensive central portion predominantly by large-caliber fibers. These fiber populations were separable on the basis of levels of α-glycerophosphate dehydrogenase and reduced nicotinamide adenine dinucleotide diaphorase activity, and to a lesser degree by glycogen content. All fibers stained strongly with the myosin adenosine triphosphatase reaction, the level of the activity being comparable to that in fast-twitch, mammalian limb muscles. The differences in features of location, size and histochemical reaction indicate that the rectus and oblique muscles in birds contain at least two types of fibers. The quadratus and pyramidalis muscles, which serve to advance the nictitating membrane, did not show segregation into zones of fibers, although in the pyramidalis two intermixed fibers were generally of larger size in the pyramidalis. Both fiber types stained darkly with myosin adenosine triphosphatase, suggesting that they were fast-twitch fibers.

The intermediate muscle fiber of rats and guinea pigs.

The soleus, plantaris and gastrocnemius muscles of 60 Sprague-Dawley rats and 70 Hartley guinea pigs were studied histochemically. In the plantaris and gastrocnemius muscles, myosin adenosine triphosphatase activity differentiated intermediate fibers from red and white fibers as determined by malate dehydrogenase and succinate dehydrogenase and reduced nicotinamide adenine dinucleotide diaphorase activities. Contrary to what is commonly reported, red fibers could not be distinguished from white fibers on the basis of myosin adenosine triphosphatase activity as is commonly reported. The intermediate fiber was characterized by minimal menadione-mediated α-glycerophosphate dehydrogenase, phosphorylase and myosin adenosine triphosphatase activity and moderate malate dehydrogenase and succinate dehydrogenase and reduced nicotinamide adenine dinucleotide diaphorase activities. It is suggested that fibers with intermediate oxidative enzyme activity are physiologically slow, white fibers are fast and red fibers are moderately slow or even fast contracting fibers.

A Comparative Histochemical Mapping of the Distribution of Acetylcholinesterase and Nicotinamide Adenine Dinucleotide–Diaphorase Activities in the Human Brain

Publisher Summary This chapter presents a comparative histochemical mapping of the distribution of acetylcholinesterase and nicotinamide adenine dinucleotide–diaphorase (NAD-d) activities in the human brain. The chapter also discusses the reliability of histochemical mappings of Ache in the brain and the probable significance of the uneven distribution of Ache in various fiber tracts. A comparison of the distribution patterns of Ache in gray matter with those of other substances presumably involved in neuronal transmission is also presented in the chapter. The presentation of the data rests on microphotographs and a description of differences in terms of more or less staining. This shortcoming has been overcome for a few histochemical methods, e.g., for NAD-d that can be quantitated by extraction and spectrophotometric determination of the amount of formazan formed in the tissue by the histochemical reaction, using small discs punched from the tissue sections. No such method is available for the histochemical quantitation of Ache activity.