WorldWideScience

Sample records for causing enzyme histochemical

  1. Enzyme histochemical studies of membrane proteases in rat subfornical organ.

    Science.gov (United States)

    De Bault, L E; Mitro, A

    1994-12-01

    Localization of membrane proteases glutamyl aminopeptidase (EAP), microsomal alanyl aminopeptidase (mAAP), dipeptidyl peptidase IV (DPP IV) and gamma-glutamyl transpeptidase (gamma-GTP) were studied in vessels of the rat subfornical organ (SFO), ependyma which cover the surface of the SFO, and adjacent brain structures. Results of enzyme histochemical reactions showed strong activity for EAP, mAAP, and gamma-GTP, but absence of DPP IV in microvessels of SFO. The ependyma which cover the SFO was positive for gamma-GTP, but negative for other studied proteases. Our results showed that the spectrum of enzymes in the majority of the vessels of SFO is similar to that of the microvessels of the adjacent brain tissue which were positive for EAP, mAAP, and gamma-GTP, but negative for DPP IV. The relative intensity of the enzyme reactions in vessels varied from central to lateral locations in the SFO and the adjacent brain tissue. There was also a difference in the relative reaction intensity from one enzyme to the other. The presence and heterogeneous distribution of the enzymes are consistent with the hypothesis that membrane proteases of the microvascular endothelium constitute an enzyme-barrier between blood and parenchyma of the SFO and between blood and brain tissue, and may be involved in metabolism or modulation of various peptides when they contact the plasma membrane of the endothelial cells of the vessels.

  2. Enzyme histochemical, biochemical and metabolic effects of Solcoseryl in connection with its radioprotective capability.

    Science.gov (United States)

    Bauer, D; Locker, A; Grigoriadis, P; Goslar, H G; Jaeger, K H

    1979-01-01

    The lethal irradiation with gamma-rays (LD 100/30) of adult mice at ambient temperature of 32.5 degrees C heavily alterates the pattern of LDH in liver. In close correspondence with the radioprotective effect of Solcoseryl on the survival rate of irradiated mice -- which amounts to 100% as compared with 40% survival of solely heat adapted animals -- Solcoseryl normalizes the LDH distribution both histochemically and biochemically. Since Solcoseryl alone generally enhances enzyme activity the normalization observed is obviously attributable to an intrinsically regulatory property of this natural compound.

  3. Biochemical and histochemical analysis of the enzyme arylsulfatase in human lesions of endodontic origin.

    Science.gov (United States)

    Aqrabawi, J; Schilder, H; Toselli, P; Franzblau, C

    1993-07-01

    Lesions of endodontic origin are areas of inflammatory response which occur as a result of untreated disease process within the root canal system. Lysosomal hydrolytic arylsulfatase A and B have been identified as major enzymes initiating and propagating bone loss by degrading chondroitin-4-sulfate. The purpose of this investigation was to examine human lesions of endodontic origin for the presence of arylsulfatase A and B. Fifteen periapical lesions were obtained at the time of periapical surgery. The lesions were analyzed for the presence of arylsulfatases using the spectrophotometer by monitoring the liberated 4-nitrocatechol at 515-nm wavelength. The same lesions were examined histochemically using the electron microscope. Five control samples from healthy periodontal ligament were evaluated in a similar manner. The results showed higher levels of arylsulfatase A in lesions than in control tissues, and marked activity of arylsulfatase B in lesions, whereas no activity of this enzyme was detected in the control specimen. Histochemically, all lesions showed positive staining for enzyme activity, whereas the controls were negative. These findings indicate that arylsulfatase A and B play a role in the pathogenesis of human lesions of endodontic origin.

  4. Histochemical studies on genetical control of hormonal enzyme inducibility in the mouse. V. Histochemical evidence for androgen inducibility of beta-glucuronidase in the epididymis

    DEFF Research Database (Denmark)

    Blecher, S R; Kirkeby, S

    1982-01-01

    The enzyme beta-glucuronidase (beta G) is shown, by histochemical methods, to be androgen inducible in the mouse epididymis. This trait has previously been believed to exist only in the kidney. Its presence in the genital tract constitutes a valuable tool in study of the developmental genetics of...... of the reproductive system. Data presented here and previously imply co-ordinated genetic control of heterogeneous lysosomal populations. The results reported also provide a system for study of X-chromosomal activation, and of the developmental androgen dependence of the epididymis....

  5. Membrane-bound proteases of the gerbil subfornical organ and choroid plexus: an enzyme histochemical study.

    Science.gov (United States)

    Mitro, A; De Bault, L E

    1994-03-01

    Using enzyme-histochemical methods, the membrane-bound peptidases, gamma-glutamyl transpeptidase (gamma-GTP), microsomal alanyl aminopeptidase (mAAP), glutamyl aminopeptidase (EAP), and dipeptidyl peptidase IV (DPP IV), were studied in microvessels of the gerbil subfornical organ (SFO), choroid plexus adjacent to the SFO, and the ependyma of brain ventricle walls in the vicinity of the SFO. Vessels and microvessels of gerbil SFO and choroid plexus were positive for gamma-GTP, mAAP, and EAP, but negative for DPP IV. Blood-brain barrier (BBB) microvessels in the surrounding brain tissue also showed positive reactions for gamma-GTP, mAAP, and EAP but a negative reaction for DPP IV. Both epithelial cells of the choroid plexus and ependymal cells of the ventricle walls were negative for all four studied enzymes. It is suggested that blood-borne peptide hormones which can be substrates for these membrane-bound proteases can be modulated by gamma-GTP, mAAP, and EAP, but not by DPP IV, when they come in contact with the plasma membrane of the endothelial cells of the vessels in gerbil SFO, choroid plexus, and surrounding brain tissue.

  6. Dynamical changing pattems of glycogen and enzyme histochemical activities in rat liver graft undergoing warm ischemia injury

    Institute of Scientific and Technical Information of China (English)

    Xiao-Shun He; Yi Ma; Lin-Wei Wu; Jin-Lang Wu; Rui-De Hu; Gui-Hua Chen; Jie-Fu Huang

    2005-01-01

    AIM: To investigate the changing patterns of glycogen and enzyme histochemical activities in rat liver graft under a dif ferent warm ischemia time (WIT) and to predict the tolerant time limitation of the liver graft to warm ischemia injury.METHODS: The rats were randomized into five groups, WTT was 0, 15, 30, 45, 60 min, respectively, and histochemical staining of liver graft specimens was observed. The recovery changes of glycogen and enzyme histochemistry activities were measured respectively 6 and 24 h following liver graft implantation.RESULTS: The activities of succinic dehydrogenase, cytochrome oxidase, apyrase (Mg++-ATPase) and content of glycogen were decreased gradually after different WIT in a time-dependent manner. The changes were significant when WIT was over 30 min.CONCLUSION: Hepatic injury is reversible within 30 min of warm ischemia injury. Glycogen and enzyme histochemistry activities of liver grafts and their recovery potency after reperfusion may serve as criteria to evaluate the quality of liver grafts.

  7. Enzyme-histochemical study on postnatal development of rat stomach lymphatic vessels.

    Science.gov (United States)

    Ji, R C; Kato, S

    1997-07-01

    Postnatal development of rat gastric lymphatics was studied by an enzyme-histochemical method to elucidate the morphological changes of lymphatics and their relationship to maturation and function, especially in the glandular portion. The significant features of 5'-Nase-positive lymphatics in distribution and structure were examined in different stages (within 24 hr, 4-21 days, and 2 months). Lymphatics in the greater curvature and anterior wall grew much slower than those in the lesser curvature and posterior wall of the stomach in newborn and infant rats. Lymphatic islands isolated from the primary lymphatic networks in the submucosa and subserosa underwent a morphological change during this early period. This is considered one of the basic steps in lymphatic development. Occurrence of lymphatic networks in the deep lamina propria indicates that development in the gastric wall is well characterized from Day 10. With further growth and modification of lymphatics, the networks in the different layers formed an extensive communication network and many lymphatic valves were found in the submucosa and subserosa. Pinocytotic vesicles, open junctions, and intraendothelial channels were frequently detected in the mucosal and submucosal lymphatic networks of the corpus-antrum and antrum-duodenum divisional zones in the adult rats. These findings suggest that developing lymphatics in the rat stomach may represent rapidly growing tissue not only with high 5'-Nase activity but also with high adaptability for future physiological demands.

  8. Biochemical and histochemical changes in energy supply enzyme pattern of muscles of the rat during old age.

    Science.gov (United States)

    Bass, A; Gutmann, E; Hanzlíková, V

    1975-01-01

    Senile muscles of the rat (28-36 months) show loss of overall activity of glycolytic and aerobic enzymes. However, there is a differential loss and shift of enzyme activity pattern in the three types of muscles. The extensor digitorum longus (EDL) and diaphragm show a decrease of ratios of glycolytic to aerobic-oxidative enzymes. This shift to a more oxidative type of metabolism is not observed in the soleus muscle. Decrease of enzyme activities is least marked in the diaphragm muscle. Biochemical analysis shows a trend to levelling out of metabolic differences between the different muscle types. This trend of 'dedifferentiation' is most marked when comparing EDL and soleus, least marked when comparing EDL and diaphragm muscle. The histochemical analysis shows a shift from the original mixed to a more uniform pattern of muscle fibres in the EDL and soleus muscle; this levelling-out of differences between enzymatic activities of different muscle fibres is not observed in the diaphragm muscle. Preferential atrophy and loss of ATPase activity in II muscle fibres in the soleus muscle and the occurrence of 'type grouping' are further characteristic features of senile muscle change. The findings show general (i.e. loss of enzyme activities) and differential trends of biochemical and histochemical enzyme changes in different types of muscles.

  9. Histochemical studies on genetical control of hormonal enzyme inducibility in the mouse. III. Beta-glucuronidase distribution pattern of epididymis in different genotypes

    DEFF Research Database (Denmark)

    Blecher, S R; Kirkeby, S

    1979-01-01

    This article reports the application of Hayashi's histochemical technique for beta-glucuronidase to mouse epididymis. A methodological study, which established optimal conditions for demonstrating the enzyme in this organ, is reported. The distribution pattern of beta-glucuronidase is described a...

  10. Histochemical studies on reserve substances and enzymes in female gametophyte of Zea mays.

    Science.gov (United States)

    Singh, M B; Malik, C P

    1976-01-01

    Cytochemical changes during the early development of maize caryopsis are reported. Changes in the localization of different reserve substances (e.g. polysaccharides, proteins, nucleic acids and lipids) and enzymes (acid phosphatase, esterase, lipase, phosphorylase, succinate dehydrogenase, cytochrome oxidase and peroxidase) have been studied in unfertilized and fertilized ovules. Before pollination very feeble enzyme activity (acid phosphatase, succinate dehydrogenase, cytochrome oxidase and peroxidase) was observed. Reserve substances were present in low amounts before pollination. Pollination stimulated the accumulation of several substances and enzymes in the tip of the nucellus, micropylar zone. Just prior to, during and after fertilization, the cells in the micropylar zone had strong reaction for several enzymes indicating temporary enhancement of metabolic activity in the micropylar zone. The role of antipodals in the storage of reserve food products and nutrition of embryo and early stages of endosperm development is discussed. The pattern of enzymatic changes within the embryo sac reflected the biochemical changes operative during quiescent and active stages. The nucellus of Zea mays contains many enzymes required for hydrolysis of reserved food substances. A role of acid phosphatase in autolysis of nucellar cells, after fertilization is suggested. Post-fertilization increase in the activity of enzymes and accumulation of reserve materials is interpreted as reflecting a presumed increase in the metabolic rate relative to growth and differentiation.

  11. Histopathological and histochemical changes in the liver of Bagrus bayad caused by environmental pollution.

    Science.gov (United States)

    Gaber, Hanan S; Ibrahim, Seham A; El-Kasheif, Midhat A

    2015-09-01

    This study focused on two areas (Abu-Zaabal and El-Tal El-Kabir) of the Ismailia canal. The samples represented the effect of the factories' effluent discharge onto the canal. Physical characteristics (water temperature and pH) and chemical characteristics (total hardness, dissolved oxygen, biological oxygen demand and chemical oxygen demand) were measured to identify the water quality in the two areas of the Ismailia canal. These measurements showed slight variations between the two stations. Additionally, the histological and histochemical analyses in the liver of Bagrus bayad fish samples were done. Histological studies of liver showed more severe degeneration in the fish samples of El-Tal El-Kabir station. Histochemical studies on protein and glycogen contents in the liver of B. bayad showed a progressive decrease in their staining intensity to bromophenol blue and periodic acid Schiff at two stations. This study, therefore, concludes that Ismailia canal water (Abu-Zaabal and El-Tall El-Kabir) stations create metabolic crisis and impairment in fish liver. The depletion in the levels of protein and glycogen points leads to the exhaustion of cell energy that is of high demand for fish during stressful conditions. Therefore, it is imperative for the authorities concerned to ensure that treated effluent discharge comply with acceptable standards to save our environment from destruction.

  12. Histochemical studies on genetical control of hormonal enzyme inducibility in the mouse. IV: Cellular localization of androgen sensitive nonspecific esterase in the epididymis

    DEFF Research Database (Denmark)

    Kirkeby, S; Blecher, S R

    1981-01-01

    cell type, is shown to be androgen dependent. When mice of different strains were castrated or treated with antiandrogens, the characteristic histochemical reaction product disappeared or was greatly reduced. Androgen treatment of castrated animals caused the reaction in the cells concerned to return...

  13. HISTOLOGICAL AND ENZYME HISTOCHEMICAL STUDIES ON THE TRANSITIONALGONADS OF GIANT GROUPER E. tauvina (PERCIFORMSSERRANIDAE) FROM THE ARABIAN GULF COAST OF SAUDI ARABIA.

    OpenAIRE

    Nora Ahmed Obaid Al-kaabi

    2010-01-01

    The present work was carried out using twenty mature Epinephelus tauvina (Perciforms: Serranidae) collected from Arabian Gulf coast at Dammam City. Fishes of this species are known to undergo sex change during certain stage of their life cycle. Histological and enzyme histochemical studies were performed on gonads of the collected fishes. The examination of the gonads of E. tauvina revealed the presence of three developmental phases during the sex change process. These were: Female, early tra...

  14. Histochemical demonstration of mercury induced changes in rat neurons

    DEFF Research Database (Denmark)

    Danscher, G; Schrøder, H D

    1979-01-01

    A histochemical method modified for ultrastructural studies of mercury induced changes is described. Rat neurons from areas known to be influenced by mercury are used as examples. The histochemical reaction, suggested to be caused by polymercury sulphide complexes, is localized to "dense bodies...

  15. Enzyme

    Science.gov (United States)

    Enzymes are complex proteins that cause a specific chemical change in all parts of the body. For ... use them. Blood clotting is another example of enzymes at work. Enzymes are needed for all body ...

  16. Histochemical analysis of experimental granulomatous uveitis.

    Science.gov (United States)

    Kristeva, M; Biswas, J; Pararajasegaram, G; Sevanian, A; Rao, N A

    1991-01-01

    We have previously demonstrated the effects of various inhibitors of arachidonic acid metabolism on experimental lens-induced granulomatous uveitis. In the present study, we investigated the effect of these same inhibitors on the expression of lysosomal enzymes at different stages of choroidal inflammation in experimental lens-induced granulomatous uveitis and compared this to the inflammation observed at each stage examined. Lysosomal enzymes such as acid phosphatase, beta-glucuronidase and succinate dehydrogenase are known to be liberated during the maturation of mononuclear phagocytes to epithelioid cell granulomas. Although animals treated with nordihydroguaiaretic acid showed less severe inflammation than did indomethacin-treated or control animals, none of these agents appeared to affect the expression of acid phosphatase and beta-glucuronidase, as determined histochemically. Succinate dehydrogenase could not be detected in any of the eyes examined, even though sections of liver and kidney from these same animals were positive for this enzyme.

  17. Mitochondrial neurogastrointestinal encephalomyopathy caused by thymidine phosphorylase enzyme deficiency: From pathogenesis to emerging therapeutic options

    NARCIS (Netherlands)

    R. Yadak (Rana); P.S. Smitt; M.W. van Gisbergen; N.P. van Til (Niek); I.F.M. de Coo (René)

    2017-01-01

    textabstractMitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a progressive metabolic disorder caused by thymidine phosphorylase (TP) enzyme deficiency. The lack of TP results in systemic accumulation of deoxyribonucleosides thymidine (dThd) and deoxyuridine (dUrd). In these patients,

  18. Asparaginase II of Saccharomyces cerevisiae: selection of four mutations that cause derepressed enzyme synthesis.

    Science.gov (United States)

    Kamerud, J Q; Roon, R J

    1986-01-01

    A positive selection method was used to isolate four Saccharomyces cerevisiae mutations that cause derepressed synthesis of asparaginase II. The four mutations (and1, and2, and3, and4) were neither closely linked to each other nor linked to previously characterized mutations (asp3, asp6) which cause the complete loss of asparaginase II activity. One of the new mutations (and4) was shown to be allelic to gdh-CR, a pleiotropic mutation which causes derepressed synthesis of a number of enzymes of nitrogen catabolism.

  19. Feline congenital erythropoietic porphyria: two homozygous UROS missense mutations cause the enzyme deficiency and porphyrin accumulation.

    Science.gov (United States)

    Clavero, Sonia; Bishop, David F; Giger, Urs; Haskins, Mark E; Desnick, Robert J

    2010-01-01

    The first feline model of human congenital erythropoietic porphyria (CEP) due to deficient uroporphyrinogen III synthase (URO-synthase) activity was identified by its characteristic clinical phenotype, and confirmed by biochemical and molecular genetic studies. The proband, an adult domestic shorthair cat, had dark-red urine and brownish discolored teeth with red fluorescence under ultraviolet light. Biochemical studies demonstrated markedly increased uroporphyrinogen I in urine and plasma (2,650- and 10,700-fold greater than wild type, respectively), whereas urinary 5-aminolevulinic acid and porphobilinogen were lower than normal. Erythrocytic URO-synthase activity was UROS gene revealed two missense mutations, c.140C>T (p.S47F) in exon 3 and c.331G>A (p.G111S) in exon 6, both of which were homozygous, presumably owing to parental consanguinity. Neither was present in 100 normal cat alleles. Prokaryotic expression and thermostability studies of the purified monomeric wild-type, p.S47F, p.G111S, and p.S47F/G111S enzymes showed that the p.S47F enzyme had 100% of wild-type specific activity but ~50% decreased thermostability, whereas the p.G111S and p.S47F/G111S enzymes had about 60% and 20% of wild-type specific activity, respectively, and both were markedly thermolabile. Molecular modeling results indicated that the less active/less stable p.G111S enzyme was further functionally impaired by a structural interaction induced by the presence of the S47F substitution. Thus, the synergistic interaction of two rare amino acid substitutions in the URO-synthase polypeptide caused the feline model of human CEP.

  20. Synergistic Degradation of a Hyperuricemia-Causing Metabolite Using One-Pot Enzyme-Nanozyme Cascade Reactions

    Science.gov (United States)

    Jung, Secheon; Kwon, Inchan

    2017-01-01

    Multi-enzyme cascade reactions are frequently found in living organisms, in particular when an intermediate should be eliminated. Recently, enzyme-mimic nanomaterials (nanozymes) received much attention for various applications, because they are usually more stable and cost-effective than enzymes. However, enzyme-nanozyme cascade reations have not been yet extensively exploited. Therefore, in this study, we investigated one-pot enzyme-nanozyme cascade reactions using urate oxidase (UOX) and catalase-mimic gold nanoparticle nanozyme (AuNP) with the ultimate goal of treatment of hyperuricemia. UOX degrades hyperuricemia-causing uric acid, but also generates hydrogen peroxide raising several health concerns. We successfully demonstrated that one-pot UOX-AuNP cascade systems degrade uric acid more than five times faster than UOX alone, by eliminating potentially cytotoxic hydrogen peroxide, similar to enzyme-enzyme reactions. PMID:28287162

  1. Ultrastructural and Histochemical Characterization of the Zebra Mussel Adhesive Apparatus

    Science.gov (United States)

    Farsad, Nikrooz

    Since their accidental introduction into the Great Lakes in mid- to late-1980s, the freshwater zebra mussels, Dreissena polymorpha, have colonized most lakes and waterways across eastern North America. Their rapid spread is partly attributed to their ability to tenaciously attach to hard substrates via an adhesive apparatus called the byssus, resulting in serious environmental and economic impacts. A detailed ultrastructural study of the byssus revealed a 10 nm adhesive layer at the attachment interface. Distributions of the main adhesive amino acid, 3,4-dihydroxyphenylalanine (DOPA), and its oxidizing (cross-linking) enzyme, catechol oxidase, were determined histochemically. It was found that, upon aging, DOPA levels remained high in the portion of the byssus closest to the interface, consistent with an adhesive role. In contrast, reduced levels of DOPA corresponded well with high levels of catechol oxidase in the load-bearing component of the byssus, presumably forming cross-links and increasing the cohesive strength.

  2. Crystal structure of glycogen debranching enzyme and insights into its catalysis and disease-causing mutations.

    Science.gov (United States)

    Zhai, Liting; Feng, Lingling; Xia, Lin; Yin, Huiyong; Xiang, Song

    2016-04-18

    Glycogen is a branched glucose polymer and serves as an important energy store. Its debranching is a critical step in its mobilization. In animals and fungi, the 170 kDa glycogen debranching enzyme (GDE) catalyses this reaction. GDE deficiencies in humans are associated with severe diseases collectively termed glycogen storage disease type III (GSDIII). We report crystal structures of GDE and its complex with oligosaccharides, and structure-guided mutagenesis and biochemical studies to assess the structural observations. These studies reveal that distinct domains in GDE catalyse sequential reactions in glycogen debranching, the mechanism of their catalysis and highly specific substrate recognition. The unique tertiary structure of GDE provides additional contacts to glycogen besides its active sites, and our biochemical experiments indicate that they mediate its recruitment to glycogen and regulate its activity. Combining the understanding of the GDE catalysis and functional characterizations of its disease-causing mutations provides molecular insights into GSDIII.

  3. First contiguous gene deletion causing biotinidase deficiency: The enzyme deficiency in three Sri Lankan children

    Directory of Open Access Journals (Sweden)

    Danika Nadeen Senanayake

    2015-03-01

    Full Text Available We report three symptomatic children with profound biotinidase deficiency from Sri Lanka. All three children presented with typical clinical features of the disorder. The first is homozygous for a missense mutation in the BTD gene (c.98_104 del7insTCC; p.Cys33PhefsX36 that is commonly seen in the western countries, the second is homozygous for a novel missense mutation (p.Ala439Asp, and the third is the first reported instance of a contiguous gene deletion causing the enzyme deficiency. In addition, this latter finding exemplifies the importance of considering a deletion within the BTD gene for reconciling enzymatic activity with genotype, which can occur in asymptomatic children who are identified by newborn screening.

  4. Effect of Hymenolepis nana on the mouse ileal mucosal cells: histochemical studies.

    Science.gov (United States)

    Sanad, M M; Salem, S A; el-Gamal, R L; Darwish, R A; el-Ridi, A M

    1990-12-01

    Histochemical studies of the ileal mucosal cells of mice experimentally infected with H. nana revealed definite increase in mucous secretions indicating increased activity of the goblet cells in response to mucosal irritation. The activity of acid phosphatase was also increased representing a sort of defence mechanism against the attached worms. The activities of ATP-ase and NADH diaphorase enzymes were decreased indicating disturbance in the metabolic and transport processes and in the absorptive function of the intestinal epithelial cells.

  5. Parvoviruses cause nuclear envelope breakdown by activating key enzymes of mitosis.

    Science.gov (United States)

    Porwal, Manvi; Cohen, Sarah; Snoussi, Kenza; Popa-Wagner, Ruth; Anderson, Fenja; Dugot-Senant, Nathalie; Wodrich, Harald; Dinsart, Christiane; Kleinschmidt, Jürgen A; Panté, Nelly; Kann, Michael

    2013-10-01

    Disassembly of the nuclear lamina is essential in mitosis and apoptosis requiring multiple coordinated enzymatic activities in nucleus and cytoplasm. Activation and coordination of the different activities is poorly understood and moreover complicated as some factors translocate between cytoplasm and nucleus in preparatory phases. Here we used the ability of parvoviruses to induce nuclear membrane breakdown to understand the triggers of key mitotic enzymes. Nuclear envelope disintegration was shown upon infection, microinjection but also upon their application to permeabilized cells. The latter technique also showed that nuclear envelope disintegration was independent upon soluble cytoplasmic factors. Using time-lapse microscopy, we observed that nuclear disassembly exhibited mitosis-like kinetics and occurred suddenly, implying a catastrophic event irrespective of cell- or type of parvovirus used. Analyzing the order of the processes allowed us to propose a model starting with direct binding of parvoviruses to distinct proteins of the nuclear pore causing structural rearrangement of the parvoviruses. The resulting exposure of domains comprising amphipathic helices was required for nuclear envelope disintegration, which comprised disruption of inner and outer nuclear membrane as shown by electron microscopy. Consistent with Ca⁺⁺ efflux from the lumen between inner and outer nuclear membrane we found that Ca⁺⁺ was essential for nuclear disassembly by activating PKC. PKC activation then triggered activation of cdk-2, which became further activated by caspase-3. Collectively our study shows a unique interaction of a virus with the nuclear envelope, provides evidence that a nuclear pool of executing enzymes is sufficient for nuclear disassembly in quiescent cells, and demonstrates that nuclear disassembly can be uncoupled from initial phases of mitosis.

  6. Parvoviruses cause nuclear envelope breakdown by activating key enzymes of mitosis.

    Directory of Open Access Journals (Sweden)

    Manvi Porwal

    2013-10-01

    Full Text Available Disassembly of the nuclear lamina is essential in mitosis and apoptosis requiring multiple coordinated enzymatic activities in nucleus and cytoplasm. Activation and coordination of the different activities is poorly understood and moreover complicated as some factors translocate between cytoplasm and nucleus in preparatory phases. Here we used the ability of parvoviruses to induce nuclear membrane breakdown to understand the triggers of key mitotic enzymes. Nuclear envelope disintegration was shown upon infection, microinjection but also upon their application to permeabilized cells. The latter technique also showed that nuclear envelope disintegration was independent upon soluble cytoplasmic factors. Using time-lapse microscopy, we observed that nuclear disassembly exhibited mitosis-like kinetics and occurred suddenly, implying a catastrophic event irrespective of cell- or type of parvovirus used. Analyzing the order of the processes allowed us to propose a model starting with direct binding of parvoviruses to distinct proteins of the nuclear pore causing structural rearrangement of the parvoviruses. The resulting exposure of domains comprising amphipathic helices was required for nuclear envelope disintegration, which comprised disruption of inner and outer nuclear membrane as shown by electron microscopy. Consistent with Ca⁺⁺ efflux from the lumen between inner and outer nuclear membrane we found that Ca⁺⁺ was essential for nuclear disassembly by activating PKC. PKC activation then triggered activation of cdk-2, which became further activated by caspase-3. Collectively our study shows a unique interaction of a virus with the nuclear envelope, provides evidence that a nuclear pool of executing enzymes is sufficient for nuclear disassembly in quiescent cells, and demonstrates that nuclear disassembly can be uncoupled from initial phases of mitosis.

  7. Histochemical stains as promising means for the laser histochemical surgery of a number of pathologies

    Science.gov (United States)

    Piruzyan, L. A.; Mikhailovskiy, Ye. M.; Piruzyan, A. L.

    1999-12-01

    The directions of laboratory and clinical studies oriented to experimental confirmation of the priority concept of `laser histochemical surgery' are presented. The goal of the studies is reproduction on experimental model of a number of pathologies (in vivo and in vitro) of the `sensitization to laser radiation by staining' effect. Testing of the histochemical stains as sensitizers to laser irradiation of their `address substrates', i.e. vitally stained intracellular structures which participate in the pathologic processes evolution is under planning. The processes include: (a) metabolic disorders in the brain cells, i.e. disseminated sclerosis; (b) generalized metabolic disorders- -mucopolysaccharidosis and collagenosises (periarteritis nodosa, rheumatism, rheumatoid arthritis, sclerodermia); (3) metabolic disorders in individual organs--vessel atherosclerosis, hypercholesterolemia, myocardial infarction, cardiosclerosis, caries and parodontosis. The conditions of the studies are detailed in the recommendations along the positions: (1) disease name; (2) disease characteristics: (a) pathomorphologic, (b) biochemical; (3) stains revealing the disease signs and recommended for testing; (4) `address substrates' of the stains that are targets for laser radiation; (5) lasers recommended for the testing after the cells staining in vivo in the corresponding pathology; (6) experimental models of the pathologies suggested for the testing; (7) criteria of the stain efficiency as target sensitizer to the laser light (criteria of the `laser sensitization by staining' efficiency). Possible perspectives for the experimental clinical medicine are indicated of common histochemical stains and lasers use and of practice introduction of the `laser histochemical surgery' in the case the described concept is confirmed in experiments and clinically.

  8. Histochemical and immunohistological approach to comparative neuromuscular diseases.

    Directory of Open Access Journals (Sweden)

    Serenella Papparella

    2009-12-01

    Full Text Available The broad category of neuromuscular diseases covers conditions that involve the weakness or wasting of the body muscles. These problems may occur in the spinal cord, the peripheral nerves or the muscle fibers. Some may be hereditary, while others are acquired. Commonly recognized conditions fall into the categories of myopathies, which are diseases of the muscle like muscular dystrophy, disorders of the junction where the nerve impulses are transmitted to the muscle like myasthenia gravis, and neuropathies, which are diseases of the peripheral nervous system. The diagnosis of most neuromuscular diseases rest on careful clinical evaluation of the patient, electromyography, the muscle biopsy, and in some instances, molecular genetic studies. Muscle biopsy, associated to histochemical and immunohistological techniques, plays a key role in diagnosis of many neuromuscular disorders. A number of morphological abnormalities of muscle can be recognized on histological stains such as haematoxylin and eosin and Engel trichrome. Histochemical techniques are essential for the study of muscle biopsies for four main reasons. First, they demonstrate the non-uniform nature of the muscle highlighting the different biochemical properties of specific fibre type and their selective involvement in certain disease processes. Second, they may show an absences of a particular enzyme. Third, an excess of a particular substrate can be demonstrated. Fourth, they may show structural changes in the muscle which would not be apparent with routine histological stains, such as the enzyme-deficient cores in central core disease "mouth-eaten" fibers, and abnormalities in the distribution of mitochondria. In some neuromuscular disorders there could be only non-specific myopathological features. However, a number of proteins, including sarcolemmal, sarcomeric, and nuclear proteins as well as enzymes with defects responsible for neuromuscular disorders, have been identified during

  9. Angiotensin I-converting enzyme mutation (Trp1197Stop causes a dramatic increase in blood ACE.

    Directory of Open Access Journals (Sweden)

    Andrew B Nesterovitch

    Full Text Available BACKGROUND: Angiotensin-converting enzyme (ACE metabolizes many peptides and plays a key role in blood pressure regulation and vascular remodeling. Elevated ACE levels may be associated with an increased risk for different cardiovascular or respiratory diseases, including asthma. Previously, a molecular mechanism underlying a 5-fold familial increase of blood ACE was discovered: Pro1199Leu substitution enhanced the cleavage-secretion process. Carriers of this mutation were Caucasians from Europe (mostly Dutch or had European roots. METHODOLOGY/PRINCIPAL FINDINGS: We have found a family of African-American descent whose affected members' blood ACE level was increased 13-fold over normal. In affected family members, codon TGG coding for Trp1197 was substituted in one allele by TGA (stop codon. As a result, half of ACE expressed in these individuals had a length of 1196 amino acids and lacked a transmembrane anchor. This ACE mutant is not trafficked to the cell membrane and is directly secreted out of cells; this mechanism apparently accounts for the high serum ACE level seen in affected individuals. A haplotype of the mutant ACE allele was determined based on 12 polymorphisms, which may help to identify other carriers of this mutation. Some but not all carriers of this mutation demonstrated airflow obstruction, and some but not all have hypertension. CONCLUSIONS/SIGNIFICANCE: We have identified a novel Trp1197Stop mutation that results in dramatic elevation of serum ACE. Since blood ACE elevation is often taken as a marker of disease activity (sarcoidosis and Gaucher diseases, it is important for clinicians and medical scientists to be aware of alternative genetic causes of elevated blood ACE that are not apparently linked to disease.

  10. Garlic attenuates histological and histochemical alterations in livers of Schistosoma mansoni infected mice.

    Science.gov (United States)

    Mahmoud, Y I; Riad, N H; Taha, H

    2016-08-01

    Interest in screening for new anti-schistosomal agents is growing because of increased concerns about resistance to and safety of praziquantel. We investigated the anti-schistosomal action of prophylactic and therapeutic doses of garlic on the histological and histochemical alterations caused by Schistosoma mansoni infection. Livers of infected mice were characterized by granulomas, periportal inflammation and fibrosis, hepatocyte vacuolation, fatty degeneration and necrosis, and hypertrophy and pigmentation of Kupffer cells. Significant depletion of carbohydrates and increased lipid vacuoles also were observed. All garlic regimens caused suppression of granuloma formation and amelioration of histological and histochemical changes; the continuous treatment protocol produced the best results. Garlic appears to be a safe and economical anti-schistosomal adjuvant for attenuating the pathogenicity of schistosomiasis.

  11. Aspergillus Galactomannan Enzyme-Linked Immunosorbent Assay Cross-Reactivity Caused by Invasive Geotrichum capitatum

    Science.gov (United States)

    Giacchino, Mareva; Chiapello, Nadia; Bezzio, Stefania; Fagioli, Franca; Saracco, Paola; Alfarano, Alda; Martini, Vincenza; Cimino, Giuseppe; Martino, Pietro; Girmenia, Corrado

    2006-01-01

    We report three cases of invasive Geotrichum capitatum infection in patients with acute leukemia for which an enzyme-linked immunosorbent assay (ELISA) for Aspergillus galactomannan was positive, with no evidence of aspergillosis. Supernatants obtained from suspensions of 17 G. capitatum strains gave positive reactions with the Aspergillus galactomannan ELISA. These clinical and laboratory data seem to suggest that G. capitatum produces a soluble antigen that is cross-reactive with Aspergillus galactomannan. PMID:16954294

  12. Phytoestrogens in milk: Overestimations caused by contamination of the hydrolytic enzyme used during sample extraction.

    Science.gov (United States)

    Bláhová, L; Kohoutek, J; Procházková, T; Prudíková, M; Bláha, L

    2016-09-01

    Isoflavones are natural phytoestrogens with antioxidant and endocrine-disrupting potencies. Monitoring of their levels is important to ensure the high quality and safety of food, milk, and dairy products. The efficiency and accuracy of phytoestrogen analyses in complex matrices such as milk depend on the extraction procedure, which often uses hydrolysis by means of the β-glucuronidase/sulfatase enzyme originating from Helix pomatia. The present study reveals that the commercially available hydrolytic enzyme is contaminated by several phytoestrogen isoflavones (genistein, daidzein, formononetin, and biochanin A) and their metabolite equol, as well as flavones (naringenin and apigenin) and coumestrol. We show that the concentrations of daidzein and genistein in the enzyme could have impaired the results of analyses of the main isoflavones in several previously published studies. Of 8 analyzed compounds, only equol was confirmed in the present study and it serves as a reliable marker of phytoestrogens originating from cow feed. Critical reassessment of phytoestrogen concentrations in milk is needed because several previously published studies might have overestimated the concentrations depending on the extraction procedure used.

  13. Histochemical demonstration of methamphetamine by immunocytochemistry.

    Science.gov (United States)

    Ishiyama, I; Mukaida, M; Yoshii, T; Suyama, H

    1987-05-01

    A method for the demonstration of methamphetamine (MA) by immunocytochemistry was established. The tissues of intoxicated mice, administered various amounts of MA in single doses of from 0.01 to 1 mg of MA-HCl, were fixed in glutaraldehyde-containing fixatives. Cryostat and paraffin slices gave a positive reaction of MA localization by staining the brain, liver, kidney, lung, stomach, spleen, and so forth, with the aid of the indirect immunoperoxidase technique. Those of animals administered a single dose of 0.1 mg or more (over 3 to 4 mg/kg--the usual dose of MA in acute intoxication death in forensic medicine), in particular, gave a strong strong reaction, so that the diagnosis of MA intoxication can be performed by macroscopic observation of stained slices. The histochemical diagnosis of MA intoxication in clinical toxicology and pathology might be regarded as a useful tool, especially in forensic pathology. The following cells gave a strong positive reaction: nerve cells and myelin sheaths, hepatocytes, epithelial cells of the distal part of the renal tubule and of the collecting tubule, alveolar and bronchial epithelial cells of the lung, chief and parietal cells of the gastric gland, capillaries of the renal glomerulus, macrophages in the blood and tissues, and striated muscle cells including cardiocytes. The morphological evidence of the pharmacodynamics and pharmacokinetics of MA can be determined at the cellular level by immunocytochemistry.

  14. Influence of liver enzyme induction caused by diphenylhydantoin and diazepam on bone mass of rats.

    Science.gov (United States)

    Tesseromatis, C; Galanopoulou, P; Varonos, D

    1980-01-01

    25 albino rats of Wistar strain were used for the study. They were divided into three groups. Animals of the first group (n = 10) were treated with Diphenylhydantoin in a daily dose of 70 mg/kg for three months. Animals of the second group (n = 10) were treated with Diazepam in a daily dose of 10 mg/kg for 3 months. Animals of the third group (n = 5) were treated with 5% acacia oil for the same period of time. Food and water consumption together with motor activity were measured. As indices of enzyme induction, D-Glucaric acid and liver weight were determined. Also the following bone indices were determined: femur Ca/femur weight, femur Ca/body weight, humerus Ca/humerus weight, humerus Ca/body weight, femur specific weight, humerus specific weight. Statistically significant alterations of bone mass were found.

  15. Physiology of sexual reproduction. I. A histochemical study of the embryo sac development in Zephyranthes rosea and Lagenaria vulgaris.

    Science.gov (United States)

    Malik, C P; Vermani, S

    1975-01-01

    Using histochemical techniques, changes in the localization of different reserve substances (e.g. pectic compounds, starch, polysaccharides, proteins, nucleic acids, ascorbic acid) and enzymes (Acid phosphatase, alkaline phosphatase, ATP-ase, 5-nucleotidase, esterase, phosphorylase, succinate dehydrogenase, cytochrome oxidase and lipase) have been studied in the young and fertilized ovules of Zephyranthes sp. and Lagenaria sp. etc. Extensive changes in the enzyme activity and reserve substances are demonstrated during megasporogenesis and megagametogenesis and most of the substances exhibited interesting distributional pattern. Similarly, all the enzymes investigated have specific locale of distribution in the tissues which displayed differentiation of embryo sac. The earlier changes observed are in the megaspore which contained many reserve substances (starch; nucleic acids; ascorbic acid; proteins) and enzymes (peroxidase, succinate dehydrogenase, acid phosphatase, alkaline phosphatase and ATP-ase). In the matured embryo sac different cells have differential localization of the substances. Based on histochemical studies, distinct differences are made out between egg and synergids; egg and central cell. In general antipodals have maximum accumulation of physiologically active substances and intense activity of different enzymes. Nucellus cells also stored diverse substances and enzymes especially towards the chalazal end. Pollination stimulated accumulation of several reserve substances and enzymes in the tip of nucellus beak, micropylar zone and these included starch, peroxidase, phosphorylase succinate dehydrogenase, cytochrome oxidase etc.

  16. A missense mutation P136L in the arylsulfatase A gene causes instability and loss of activity of the mutant enzyme.

    Science.gov (United States)

    Kafert, S; Heinisch, U; Zlotogora, J; Gieselmann, V

    1995-02-01

    Metachromatic leukodystrophy is a lysosomal storage disease caused by deficiency of arylsulfatase A. Sequencing of the arylsulfatase A genes of an Ashkenazi Jewish patient suffering from the severe late infantile form of the disease revealed a point mutation in exon 2 causing proline 136 to be substituted by leucine. The patient was homozygous for this mutation. Studies on Ltk- cells stably expressing the mutant enzyme show that the mutation causes complete loss of enzyme activity and rapid degradation in an early biosynthetic compartment.

  17. Knockout of Lysosomal Enzyme-Targeting Gene Causes Abnormalities in Mouse Pup Isolation Calls

    Science.gov (United States)

    Barnes, Terra D.; Holy, Timothy E.

    2017-01-01

    Humans lacking a working copy of the GNPTAB gene suffer from the metabolic disease Mucolipidosis type II (MLII). MLII symptoms include mental retardation, skeletal deformities and cartilage defects as well as a speech delay with most subjects unable to utter single words (Otomo et al., 2009; Cathey et al., 2010; Leroy et al., 2012). Here we asked whether mice lacking a copy of Gnptab gene exhibited vocal abnormities. We recorded ultrasonic vocalizations from 5 to 8 day old mice separated from their mother and littermates. Although Gnptab−/− pups emitted a similar number of calls, several features of the calls were different from their wild type littermates. Gnptab−/− mice showed a decrease in the length of calls, an increase in the intra-bout pause duration, significantly fewer pitch jumps with smaller mean size, and an increase in the number of isolated calls. In addition, Gnptab−/− mice vocalizations had less power, particularly in the higher frequencies. Gnptab+/− mouse vocalizations did not appear to be affected. We then attempted to classify these recordings using these features to determine the genotype of the animal. We were able to correctly identify 87% of the recordings as either Gnptab−/− or Gnptab+/+ pup, significantly better than chance, demonstrating that genotype is a strong predictor of vocalization phenotype. These data show that deletion of genes in the lysosomal enzyme targeting pathway affect mouse pup isolation calls.

  18. HISTOLOGICAL AND HISTOCHEMICAL CHARACTERISTICS OF RAT MYOCARDIUM IN CADMIUM TOXICOSIS

    Directory of Open Access Journals (Sweden)

    Novica Bojanić

    2013-01-01

    Full Text Available Carcinogenic effects of cadmium on lungs, testicles and prostate are well known, so as cumulative and toxic effects on kidney, liver and bones; however, there have not been many published articles about the effects of cadmium on myocardium. The aim of this study was to estimate the morphological changes in rat myocardium chronically treated by cadmium. The study was carried out on male albino Wistar rats (n=30, age=35-37 days, body mass 120g +/- 10g. The animals were raised in controlled laboratory conditions and provided with standard laboratory rat food and tap water ad libitum. The rats were divided into two groups: ten animals composed the control group and did not undergo any treatment. The 20 experimental rats were exposed to 10mg of CdCl2 /L drinking water for 90 days. After 90 days, all animals were victimized and after the macroscopic inspection of the heart, myocardial tissue was routinely processed and embedded in paraffin. Sections 5 micrometers thick were stained by HE method and histochemical PAS-AB (pH 2, 5, Masson trichrome method for demonstrating collagen fibers and Toluidine blue for mast cells identification. Cross-striated banding pattern of cardiac cells was ruined. Noticeable atrophy and hydropic degeneration of subendocardial localized cardiac cells were found, with the focal presence of myocytolysis. Endothelial cell hyperplasia and edema of the intima were present on arteriolar type blood vessels causing the focal subocclusion. Fibrocytes, histiocytes and mast cells were numerous, perivascularly localized. Mast cells were polymorphic, larger than normal, oval and mostly degranulated. Instead of scanty endomysium, there is a noticeable interstitial fibrillar fibrosis with few fields of collagen in all myocardium layers between cardiac cells, which is particularly prominent around the larger blood vessels. Cadmium has pronounced vasculotropic properties causing morphological changes of cardiomyocytes, myocardial

  19. PRELIMINARY PHYTOCHEMICAL AND HISTOCHEMICAL INVESTIGATION ON KIGELIA PINNATA DC.,

    Directory of Open Access Journals (Sweden)

    DHANASEKARAN.M

    2014-07-01

    Full Text Available Kigelia pinnata D.C. the mid sized ornamental tree of the Bignoniaceae has been studied by preliminary phytochemical and histochemical analysis. Several local names are availability to this plants based on their country. They are called worsboom in Africa and sauage tree in America. The tree is 25 meters in hight with a dense rounded crown bark grey. Data gathered on solvent extraction and preliminary phytochemical method suggested that the presence of glycosides, flavonoides, tannin and alkaloids in leaf tissue. Anatomical and histochemical investigation offered some clues on the localization of certain specific metabolites. This paper revealed preliminary phytochemical constituents of Kigelia pinnta D.C.., by phytochemical and histochemical investigation.

  20. [Food intolerances caused by enzyme defects and carbohydrate malassimiliations : Lactose intolerance and Co].

    Science.gov (United States)

    Schäfer, Christiane

    2016-06-01

    Apart from allergic conditions, carbohydrate malassimiliations (sugar metabolism disorders) are classified within the group of food intolerances. These dose-dependent, yet non-immunological reactions require gastroenterological or internal diagnosis following nutritional therapy. Intolerances to carbohydrates such as lactose (milk sugar) and fructose (fruit sugar) in addition to sugar alcohols (sorbitol, mannitol, lactitol etc.) have been gaining increasing attention in recent decades as they are the cause of a wide range of gastrointestinal symptoms. There are currently various options for both diagnosis and therapy that differ notably in terms of effort, costs, and efficiency. Nutritional change and patient education are the bases of therapy. Non-observance of the trigger will result in increasing complaints and possibly even more infections, e.g., diverticula, rectal disorders, bacterial miscolonization, bile acid malabsorption). For an optimal therapy, the following sugar metabolism disorders have to be differentiated: hypolactasia versus lactose maldigestion, fructose malabsorption versus fructose overload, combined lactose and fructose intolerance, and isolated adverse reactions against sorbitol.For the medical conditions listed above, a three- or four-stage treatment regimen is recommended. Extensive dietary restrictions with regard to the relevant sugar, except for lactose, should not be maintained over a longer period of time.

  1. Histo-chemical and biochemical analysis reveals association of er1 mediated powdery mildew resistance and redox balance in pea.

    Science.gov (United States)

    Mohapatra, Chinmayee; Chand, Ramesh; Navathe, Sudhir; Sharma, Sandeep

    2016-09-01

    Powdery mildew caused by Erysiphe pisi is one of the important diseases responsible for heavy yield losses in pea crop worldwide. The most effective method of controlling the disease is the use of resistant varieties. The resistance to powdery mildew in pea is recessive and governed by a single gene er1. The objective of present study is to investigate if er1 mediated powdery mildew resistance is associated with changes in the redox status of the pea plant. 16 pea genotypes were screened for powdery mildew resistance in field condition for two years and, also, analyzed for the presence/absence of er1 gene. Histochemical analysis with DAB and NBT staining indicates accumulation of reactive oxygen species (ROS) in surrounding area of powdery mildew infection which was higher in susceptible genotypes as compared to resistant genotypes. A biochemical study revealed that the activity of superoxide dismutase (SOD) and catalase, enzymes involved in scavenging ROS, was increased in, both, resistant and susceptible genotypes after powdery mildew infection. However, both enzymes level was always higher in resistant than susceptible genotypes throughout time course of infection. Moreover, irrespective of any treatment, the total phenol (TP) and malondialdehyde (MDA) content was significantly high and low in resistant genotypes, respectively. The powdery mildew infection elevated the MDA content but decreased the total phenol in pea genotypes. Statistical analysis showed a strong positive correlation between AUDPC and MDA; however, a negative correlation was observed between AUDPC and SOD, CAT and TP. Heritability of antioxidant was also high. The study identified few novel genotypes resistant to powdery mildew infection that carried the er1 gene and provided further clue that er1 mediated defense response utilizes antioxidant machinery to confer powdery mildew resistance in pea.

  2. Determination of in Histochemical Changes of Internal Organs Under Acute Alcoholic Intoxications in Russia

    Directory of Open Access Journals (Sweden)

    Vugar Mammadov

    2011-12-01

    Full Text Available AIM: During forensic-medical autopsies experts must very often assess degree and the role of acute alcohol intoxications in mechanisms of death. This becomes more actual in the cases of sudden death from different diseases especially cardiac, brain and liver origin, mechanical injuries, mechanical asphyxia etc. METHOD: This study aims to show that blood concentration of alcohol must not be only criteria during assessment of severity of intoxications and their influence on patogenesis. Epidemiological screening of 320 medico-legal reports assesses the role of sex, age, ethnicity and other peculiarities. RESULTS: Special histochemical investigations of ethanol oxidizing enzymes were also conducted on materials taken from 167 autopsies. CONCLUSION: Study suggests that morpho-functional histochemical tests must be entered in the list of methods of postmortem diagnostics during autopsies of cases with the background of ethanol poisoning, especially for differential diagnosis of acute alcohol intoxication, alcohol cardiomyopathy and ischemic heart disease. [TAF Prev Med Bull 2011; 10(6.000: 641-648

  3. Correlation between histochemically assessed fiber type distribution and isomyosin and myosin heavy chain content in porcine skeletal muscles.

    Science.gov (United States)

    Bee, G; Solomon, M B; Czerwinski, S M; Long, C; Pursel, V G

    1999-08-01

    Highly sensitive enzyme assays developed to differentiate skeletal muscle fibers allow the recognition of three main fiber types: slow-twitch oxidative (SO), fast-twitch oxidative glycolytic (FOG), and fast-twitch glycolytic (FG). Myosin, the predominant contractile protein in mammalian skeletal muscle, can be separated based on the electrophoretic mobility under nondissociating conditions into SM2, SM1, IM, FM3, and FM2 isoforms, or under dissociating conditions into myosin heavy chain (MHC) I, IIb, IIx/d, and IIa. The purpose of the present study was to determine whether the histochemical method of differentiation of fiber types is consistent with the electrophoretically identified isomyosin and MHC isoforms. These comparisons were made using serratus ventralis (SV), gluteus medius (GM), and longissimus muscles (LM) from 13 pigs. Two calculation methods for the histochemical assessed fiber type distribution were adopted. The first method incorporated the number of fibers counted for each fiber type and calculated a percentage of the total fiber number (fiber number percentage: FNP). The second method expressed the cross-sectional area of each fiber type as a percentage of the total fiber area measured per muscle (fiber area percentage: FAP). Independent of the calculation methods, correlation analyses revealed in all muscles a strong relation between SO fibers, the slow isomyosin (SM1 and SM2), and MHCI, as well as between the FG fibers, the fast isomyosin (FM3 and FM2), and MHCIIx/b content (PFOG fiber population assessed by histochemical analysis and intermediate isoform (IM) or MHCIIa content. The present results did not provide conclusive evidence as to which of the calculation methods (FNP or FAP) was more closely related to myosin composition of skeletal muscles. Despite some incompatibility between the methods, the present study shows that histochemical as well as electrophoretic analyses yielded important information about the composition of porcine

  4. Histochemical study of the myenteric plexus following gastric resection on dogs.

    Science.gov (United States)

    Lolova, I; Atanassova, E

    1982-01-01

    Enzyme histochemical methods were used to follow up to changes within the myenteric plexus from 1 week to 7 months following subtotal Billroth I gastric resection in dogs. Different regions of the resected stomach were examined. The changes affected the neurons and the neuropil of the myenteric ganglia, the connecting strands and the nerve bundles and fibers within the muscle layers. The changes in size, from and enzyme activity of some myenteric neurons correspond to those observed during axon regeneration. The 2nd week showed a significant reduction of the nerve cell number in most of the examined regions of the resected stomach. Along with, the amitotic forms of cell division and immature nerve cells became more numerous. The enzyme activity of the latter increased with the time elapsed after resection, 7 months afterwards, the size of the myenteric ganglia in the resected stomach did not differ from those in the control dogs. There were some hypertrophic nerve cells with higher enzyme activity. Some changes presume the formation of new intramural ganglia. The changes in the myenteric neurons were accompanied by reactive changes in the non-neuronal elements. Hyperinnervation was observed in the gastric muscles.

  5. Histological and Histochemical Investigations of Myristica fragrans Houtt. (Myristicaceae

    Directory of Open Access Journals (Sweden)

    N.Parimala

    2013-01-01

    Full Text Available Myristica fragrans Houtt. of Myristicaceae yields the nut-mug and mace which are credited with high flavours and fragrance. They are used in all parts of the world for both as food flavours and for medicinal uses. The present study provides comprehensive microscopic details of the fruit and histochemical localization of the aroma compounds. The study fills the lacuma found in the account of previous investigations

  6. Mineralization of human premolar occlusal fissures: a quantitative histochemical microanalysis

    OpenAIRE

    Campos, Antonio; Rodriguez, I. A.; Sanchez-Quevedo, M.C.; García, J. M.; Nieto-Albano, O.H.; Gómez de Ferraris, M. E.

    2000-01-01

    The mechanisms of cariogenesis in occlusal fissures remain elusive because of limited information about fissure structure and wall mineralization. The purpose of the present study was to determine the correlation between morphological patterns in occlusal fissures in human premolars and quantitative histochemical patterns of mineralization in the walls of these formations. We used scanning electron microscopy and quantitative X-ray microanalysis with the peak-t...

  7. PHARMACOGNOSTICAL, PHYSICOCHEMICAL AND HISTOCHEMICAL EVALUATION OF BRIHAT PANCHAMOOLA CHURNA

    OpenAIRE

    Junjarwad A.V; Vyas M.K.; Harisha CR; Shukla VJ

    2011-01-01

    Brihatpanchamoola are among the most important drugs in Ayurveda, due to their wide range of therapeutic actions. To meet the enormously increasing demand, the procuring has been boosted significantly. This indeed resulted to the inclusion of the brihatpanchamoola in red listed plants. Hence in the present study pharmacognostical, physicochemical and histochemical evaluation was made to find out whether aerial parts can be used in place of roots with the reference of API. The pharmacognostica...

  8. PRELIMINARY PHYTOCHEMICAL AND HISTOCHEMICAL INVESTIGATION ON KIGELIA PINNATA DC.,

    OpenAIRE

    Dhanasekaran, M.; ABRAHAM.G.C; Mohan, S.

    2014-01-01

    Kigelia pinnata D.C. the mid sized ornamental tree of the Bignoniaceae has been studied by preliminary phytochemical and histochemical analysis. Several local names are availability to this plants based on their country. They are called worsboom in Africa and sauage tree in America. The tree is 25 meters in hight with a dense rounded crown bark grey. Data gathered on solvent extraction and preliminary phytochemical method suggested that the presence of glycosides, flavonoides, tannin and alka...

  9. Is there still room for novelty, in histochemical papers?

    Directory of Open Access Journals (Sweden)

    Carlo Pellicciari

    2016-12-01

    Full Text Available Histochemistry continues to be widely applied in biomedical research, being nowadays mostly addressed to detect and locate single molecules or molecular complexes inside cells and tissues, and to relate structural organization and function at the high resolution of the more advanced microscopical techniques. In the attempt to see whether histochemical novelties may be found in the recent literature, the articles published in the European Journal of Histochemistry in the period 2014-2016 have been reviewed. In the majority of the published papers, standardized methods have been preferred by scientists to make their results reliably comparable with the data in the literature, but  many papers (approximately one fourth of the published articles described novel histochemical methods and procedures.  It is worth noting that there is a growing interest for minimally-invasive in vivo techniques (magnetic resonance imaging, autofluorescence spectroscopy which may parallel conventional  histochemical analyses to obtain information not only on the morphological features of living organs and tissues, but also on their functional, biophysical and molecular characteristics. Thanks to this unceasing methodological refinement, histochemistry will continue to provide innovative applications in the biomedical field.

  10. Histochemical approaches to assess cell-to-cell transmission of misfolded proteins in neurodegenerative diseases

    Science.gov (United States)

    Natale, G.; Pompili, E.; Biagioni, F.; Paparelli, S.; Lenzi, P.; Fornai, F.

    2013-01-01

    Formation, aggregation and transmission of abnormal proteins are common features in neurodegenerative disorders including Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, and Huntington's disease. The mechanisms underlying protein alterations in neurodegenerative diseases remain controversial. Novel findings highlighted altered protein clearing systems as common biochemical pathways which generate protein misfolding, which in turn causes protein aggregation and protein spreading. In fact, proteinaceous aggregates are prone to cell-tocell propagation. This is reminiscent of what happens in prion disorders, where the prion protein misfolds thus forming aggregates which spread to neighbouring cells. For this reason, the term prionoids is currently used to emphasize how several misfolded proteins are transmitted in neurodegenerative diseases following this prion-like pattern. Histochemical techniques including the use of specific antibodies covering both light and electron microscopy offer a powerful tool to describe these phenomena and investigate specific molecular steps. These include: prion like protein alterations; glycation of prion-like altered proteins to form advanced glycation end-products (AGEs); mechanisms of extracellular secretion; interaction of AGEs with specific receptors placed on neighbouring cells (RAGEs). The present manuscript comments on these phenomena aimed to provide a consistent scenario of the available histochemical approaches to dissect each specific step. PMID:23549464

  11. Effect of reserpine on the histochemical and biochemical properties of rat intestinal mucin

    Energy Technology Data Exchange (ETDEWEB)

    Forstner, J.; Roomi, N.; Khorasani, R.; Kuhns, W.; Forstner, G. (Hospital for Sick Children, Toronto, (Canada))

    1991-04-01

    Biochemical and histochemical parameters of intestinal mucins were examined in control and reserpine-treated rats. An assay for intestinal mucin sulfotransferase was developed and the activity shown to increase 3.4 times over control levels in rats given intraperitonal reserpine (0.5 mg/kg body wt) daily for 7 days. Histochemical staining of intestinal sections revealed an increase in sulfomucins in goblet cells of reserpine-treated rats. The effects were prominent as early as 1 day following injection, particularly in the distal third of the small intestine, and during the next 6 days these changes spread progressively to the middle and proximal thirds. After 3 days of treatment mucins were purified from each intestinal segment and compared to control mucins with respect to composition and (35S)NaSO{sub 4} incorporation. Although individual amino acid and carbohydrate molar ratios were unchanged, the total carbohydrate and sulfate content of mucins in treated animals was elevated (two to three times above control) in the middle and distal thirds of the intestine. In vivo ({sup 35}S)SO{sub 4} incorporation into these mucins was also proportionaltely elevated, and was targetted to O-linked oligosaccharide side chains. These findings are consistent with an action of reserpine causing an increased production of mucin which is enriched in glycoprotein components bearing sulfated oligosaccharide chains. The relevance of these findings to the production of hypersulfated and hyperglycosylated mucins in cystic fibrosis is discussed.

  12. Histochemical approaches to assess cell-to-cell transmission of misfolded proteins in neurodegenerative diseases

    Directory of Open Access Journals (Sweden)

    G. Natale

    2013-03-01

    Full Text Available Formation, aggregation and transmission of abnormal proteins are common features in neurodegenerative disorders including Parkinson’s disease, Alzheimer’s disease, amyotrophic lateral sclerosis, and Huntington’s disease. The mechanisms underlying protein alterations in neurodegenerative diseases remain controversial. Novel findings highlighted altered protein clearing systems as common biochemical pathways which generate protein misfolding, which in turn causes protein aggregation and protein spreading. In fact, proteinaceous aggregates are prone to cell-to-cell propagation. This is reminiscent of what happens in prion disorders, where the prion protein misfolds thus forming aggregates which spread to neighbouring cells. For this reason, the term prionoids is currently used to emphasize how several misfolded proteins are transmitted in neurodegenerative diseases following this prion-like pattern. Histochemical techniques including the use of specific antibodies covering both light and electron microscopy offer a powerful tool to describe these phenomena and investigate specific molecular steps. These include: prion like protein alterations; glycation of prion-like altered proteins to form advanced glycation end-products (AGEs; mechanisms of extracellular secretion; interaction of AGEs with specific receptors placed on neighbouring cells (RAGEs. The present manuscript comments on these phenomena aimed to provide a consistent scenario of the available histochemical approaches to dissect each specific step.

  13. Morphological and histochemical observations of the organic components of ostrich eggshell

    Directory of Open Access Journals (Sweden)

    P.D.G. Richards

    2002-07-01

    Full Text Available The organic component of the avian eggshell can be divided into 3 portions, the shell membranes, the matrix and the cuticle. These have been well characterised in the chicken but little has been published with regard to the ostrich (Struthio camelus. A number of recent studies have indicated that the cause of intra-shell embryonic deaths in the ostrich is similar to intra-shell embryonic deaths that occur in the chicken. These deaths in the chicken are associated with the loss of or damage to the waxy cuticle and other organic components of the eggshell, which is reported to be absent in the ostrich eggshell. In this study, preliminary morphological and histochemical analyses, at the level of the light and electron microscope, have characterised the various organic components of the ostrich eggshell. The results of the histochemical and electron microscopical analyses suggest that there mayonly be 1 shell membrane in this species, which could play a major role in the limitation of bacterial penetration to the embryonic chamber. The shell membrane has a distinct elemental profile as determined by EDSanalysis. The matrix is shown to decrease in mesh size from the mammillary layer to the vertical crystal layer. The closer packing of the mesh may indicate the presence of a morphologically discernible termination signal to calcification or the remnants of an evolutionary calcified cuticle. The matrix of the pores may also forma defensive barrier against bacterial invasion, which could be damaged as a result of dipping the eggs before incubation.

  14. Enzyme kinetics approach to assess biocatalyst inhibition and deactivation caused by [bmim][Cl] ionic liquid during cellulose hydrolysis.

    Science.gov (United States)

    Nemestóthy, Nándor; Megyeri, Gábor; Bakonyi, Péter; Lakatos, Patrik; Koók, László; Polakovic, Milan; Gubicza, László; Bélafi-Bakó, Katalin

    2017-04-01

    The aim of this work was to study the inhibition and deactivation of commercial enzyme cocktail (Cellic® Htec2) in the presence of [bmim][Cl] ionic liquid employing model cellulosic substrate, carboxymethyl cellulose (CMC). It turned out from the experiments - relying on enzyme kinetics approach - that [bmim][Cl] could act as a competitive inhibitor. Furthermore, depending on the process conditions i.e. contact of enzyme solution with high concentration [bmim][Cl], severe biocatalyst inactivation should be also taken into account as a potential risk during the enzymatic cellulose hydrolysis even in as short process times as few minutes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Hepatic Rupture Caused by Hemolysis, Elevated Liver Enzyme, and Low Platelet Count Syndrome: A Case Report with Computed Tomographic and Conventional Angiographic Findings

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Cheong Bok; Ahn, Jae Hong; Choi, Soo Jung; Lee, Jong Hyeog; Park, Man Soo; Jung, Seung Mun; Ryu, Dae Sik [Dept. of Radiology, Asan Foundation, Gangneung Asan Hospital, University of Ulsan College of Medicine, Gangneung (Korea, Republic of)

    2013-03-15

    The authors recently obtained successful clinical outcome after embolization of the hepatic artery and right inferior phrenic artery in a pregnant patient with hemolysis, elevated liver enzyme, and low platelet count (HELLP) syndrome causing hepatic rupture. We report the computed tomographic and conventional angiographic findings in a case of HELLP syndrome, resulting in hepatic infarction and rupture with active bleeding.

  16. Structural And Histochemical Changes Of Albino Rat Kidney Under The Effect Of Injectable Contraceptive

    Directory of Open Access Journals (Sweden)

    Mamdouh A. Ghali

    2006-03-01

    Full Text Available The choice of safe and effective method for fertility control still under continuous search. So, discovery of structures having long duration of action which made administration by injection was an attractive alternative to oral contraceptives. Medroxyprogesterone acetate emerged from this early work as promising injectable long ­ acting contraceptive with minimal risk. This work was planned to evaluate the structural and histochemical changes induced by injectable contraceptive Depo-provera (MPA, on the kidney of adult female Albino rats as well as testing the degree of reversibility of changes that may develop after the arrest of its use. Thirty adult female Albino rats were used in this work and divided into three equal groups. Group I was used as a control, group II was intramuscularly injected with MPA 4 times (2.7 mg / rat every 3 oestrus cycles and sacrificed one day after arrest of the injection, while , group III the animals were injected with MPA by the same dose and sacrificed 30 days after arrest of the injection. The abdominal aorta was exposed and Indian ink injection was injected to study the renal vascular changes. The animals were sacrificed, the kidney was dissected and paraffin sections were prepared and stained by haematoxylin and eosin and PAS technique to study the microscopic structure and the distribution of PAS+ve materials respectively. Frozen sections were prepared and stained by both Gomori and Nachla's techniques to study the activity of acid phosphatase enzyme and succinic dehydrogenase enzyme respectively. The obtained data were statistically analyzed using Student's t.test. The injected groups showed atrophy of tubular epithelium, dilatation of tubular lumina. All recovery groups were nearly similar to normal state except PAS+ve material of renal tubules which were nearly similar to injected groups. The treated groups showed significant increase in vascular distribution and PAS+ve materials. While, non

  17. Histological and histochemical observations on the foot glands in some byssus-bearing bivalves of the Waltair coast.

    Science.gov (United States)

    Banu, A; Shyamasundari, K; Rao, K H

    1980-09-01

    The glands responsible for the formation of the byssus threads in Arca symmetrica, Barbatia obliquata and Septifer bilocularis are the white gland, phenol gland and enzyme gland. Besides these, mucous glands are also present in the sub-epithelia. The size and shape of the cells of these glands vary in one and the same species. From histochemical investigations it has been revealed that these glands contain 1,2-glycol groups in addition to disulphides and sulphhydryls. The white gland secretes a protein material and the phenol gland is rich in phenols. These two combine to form a phenolic protein which is acted upon by a polyphenol oxidase secreted by the enzyme gland and leads to the formation of a byssus thread. The mucous gland cells secrete acid mucopolysaccharides, neutral mucins and glycoproteins.

  18. Glucagon-like peptide-1 receptor signaling in acinar cells causes growth dependent release of pancreatic enzymes

    DEFF Research Database (Denmark)

    Albrechtsen, Nicolai Jacob Wewer; Albrechtsen, Reidar; Bremholm, l;

    2016-01-01

    Incretin-based therapies are widely used for type 2 diabetes and now also for obesity, but they are associated with elevated plasma levels of pancreatic enzymes and perhaps a modestly increased risk of acute pancreatitis. However, little is known about the effects of the incretin hormone glucagon...

  19. Histochemical analysis of carbohydrate moieties and sugar-specific acceptors in the kidneys of the laboratory mouse and the golden spiny mouse (Acomys russatus).

    Science.gov (United States)

    Coppee, I; Gabius, H J; Danguy, A

    1993-10-01

    The aims of this work were to histochemically compare the pattern of lectin binding and endolectin expression in different portions of nephrons of two rodent species producing either normal hyperosmotic urine (the laboratory mouse) or highly concentrated urine (Acomys russatus, the golden spiny mouse). A panel of biotinylated lectins and neoglycoproteins and the avidin-biotin-peroxidase complex technique were used on Bouin's fixed, paraffin-embedded sections. Various segments of the uriniferous tubule in both species showed differential affinity for labelled lectins and neoglycoproteins. Significant differences were also evident between comparable tubular segments in laboratory and golden spiny mouse kidneys. Whether the histochemical expression of sugar moieties of glycoconjugates as well as endolectins, thus both sides of presumed protein-carbohydrate interactions, may be correlated to the various glycoproteins which would include constituents of the glycocalyx and domains of a variety of transport enzymes deserves further studies.

  20. Histochemical alterations in colorectal carcinoma and adenoma in Egyptian patients

    Institute of Scientific and Technical Information of China (English)

    Saber A Sakr; Moshira M Abdel-Wahed; Asmaa G Abdou; Eman K El-Adely

    2016-01-01

    Objective:To evaluate the histochemical alterations inDNA and total carbohydrates, in colorectal cancer cells. Methods:This study was carried out on 48 colorectal carcinoma and 10 adenoma specimens. Hematoxylin and Eosin staining was carried out for histopathological examination to confirm the diagnosis and to evaluate the histopathological characteristics of tumor. Histologic grade and pathologic stage was assessed according to TNM staging system. Staging was also assessed according to original Dukes’ staging system.DNA was demonstrated by Feulgen method and carbohydrates were demonstrated by periodic acid Schiff’s reaction. Results:Adenoma cases showed that the cells lining the glands of the polyp have more crowded, irregular and darker nuclei (hyperchromatic), anisonucleosis, abnormal mitotic figures with prominent nucleoli and variability in the size and shape of nuclei. Colorectal carcinoma cases showed a condensation and reduction in the size of a cell nucleus associated with hyperchromatosis, pyknotic nuclei, abnormal mitotic figures, anisonucleosis, irregular nuclear membrane and inequality in the size of the nuclei (Pleomorphosis). There was a statistical significant differences between adenoma and carcinoma regarding number of mitotic cells (P = 0.03) that was in favour of malignant group. Adenoma and colorectal carcinoma cases showed periodic acid Schiff’s reactivity with different degree. Conclusions:These histochemical alterations can be so characteristic of a given tumor type and stage that they are used in cancer diagnosis and might also be related to the altered functional properties of cancer cells.

  1. Histochemical alterations in colorectal carcinoma and adenoma in Egyptian patients

    Directory of Open Access Journals (Sweden)

    Saber A Sakr

    2016-01-01

    Full Text Available Objective: To evaluate the histochemical alterations in DNA and total carbohydrates, in colorectal cancer cells. Methods: This study was carried out on 48 colorectal carcinoma and 10 adenoma specimens. Hematoxylin and Eosin staining was carried out for histopathological examination to confirm the diagnosis and to evaluate the histopathological characteristics of tumor. Histologic grade and pathologic stage was assessed according to TNM staging system. Staging was also assessed according to original Dukes’ staging system. DNA was demonstrated by Feulgen method and carbohydrates were demonstrated by periodic acid Schiff’s reaction. Results: Adenoma cases showed that the cells lining the glands of the polyp have more crowded, irregular and darker nuclei (hyperchromatic, anisonucleosis, abnormal mitotic figures with prominent nucleoli and variability in the size and shape of nuclei. Colorectal carcinoma cases showed a condensation and reduction in the size of a cell nucleus associated with hyperchromatosis, pyknotic nuclei, abnormal mitotic figures, anisonucleosis, irregular nuclear membrane and inequality in the size of the nuclei (Pleomorphosis. There was a statistical significant differences between adenoma and carcinoma regarding number of mitotic cells (P = 0.03 that was in favour of malignant group. Adenoma and colorectal carcinoma cases showed periodic acid Schiff’s reactivity with different degree. Conclusions: These histochemical alterations can be so characteristic of a given tumor type and stage that they are used in cancer diagnosis and might also be related to the altered functional properties of cancer cells.

  2. Structural and Histochemical Characterization of Developing Rice Caryopsis

    Institute of Scientific and Technical Information of China (English)

    YU Xu-run; ZHOU Liang; XIONG Fei; WANG Zhong

    2014-01-01

    The development of pericarp, seed coat, starchy endosperm and aleurone of the rice caryopsis was investigated, histochemically and structurally, from the time of flowering to maturity. The results showed that during its growth, the maximum length of the caryopsis was attained first, followed by width and then thickness. Histochemical examination of the caryopsis showed that starch was mainly accumulated in the endosperm, but the endosperm showed no metabolic activity, while embryo and pericarp contained a few starch grains, and embryo and aleurone were strongly active. Aleuronic cells contained many aleurone grains and spherosomes, and aleurone in the dorsal region developed earlier and contained more layers of cells. Amyloplasts in endosperm contained many starch granules and were spherical at early stages but polyhedric at late stages. The protein bodies appeared later than amyloplasts, and the number of protein bodies in subaleurone was greater than those in the starchy endosperm. The white-belly portion of endosperm might be relative to the status of amyloplast development.

  3. Allosteric transition and binding of small molecule effectors causes curvature change in central β-sheets of selected enzymes.

    Science.gov (United States)

    Tolonen, Ellen; Bueno, Brenda; Kulshreshta, Sanjeev; Cieplak, Piotr; Argáez, Miguel; Velázquez, Leticia; Stec, Boguslaw

    2011-04-01

    A quantitative description of allosteric transition remains a significant science challenge. Many allosteric enzymes contain a central β-sheet in their catalytic domain. When an allosteric protein undergoes the transition between T (tense) and R (relaxed) allosteric states, this central β-sheet undergoes a conformational change. A traditional method of measuring this change, the root mean square deviation (RMSD), appears to be inadequate to describe such changes in meaningful quantitative manner. We designed a novel quantitative method to demonstrate this conformational transition by measuring the change in curvature of the central β-sheet when enzymes transition between allosteric states. The curvature was established by calculating the semiaxes of a 3-D hyperboloid fitted by least squares to the Cα atomic positions of the β-sheet. The two enzymes selected for this study, fructose 1,6-bisphosphatase (FBPase) from pig kidney and aspartate carbamoyltransferase (ATCase) from E. coli, showed while transitioning between the allosteric states (T ⇔ R) a notable change in β-sheet curvature (∼5%) that results in a large lateral shift at the sheet's edge, which is necessary to convey the signal. The results suggest that the β-sheet participates in storing elastic energy associated with the transition. Establishing a tentative link between the energetics of the β-sheet in different allosteric states provides a more objective basis for the naming convention of allosteric states (tense or relaxed), and provides insight into the hysteretic nature of the transition. The approach presented here allows for a better understanding of the internal dynamics of allosteric enzymes by defining the domains that directly participate in the transition.

  4. Histochemical aspects of wheat resistance to leaf blast mediated by silicon

    Directory of Open Access Journals (Sweden)

    Washington Luís da Silva

    2015-08-01

    Full Text Available Blast, caused by Pyricularia oryzae, has become a significant disease threat to wheat (Triticum aestivum L. in Brazil. This study aimed to investigate at the histochemical level if silicon (Si could enhance the production of flavonoids in the leaves of wheat plants in response to P. oryzae infection. Plants from the Aliança cultivar, which are susceptible to blast, were grown in hydroponic cultures containing 0 (-Si or 2 mM of Si (+Si and inoculated by spraying a conidial suspension of P. oryzae (1 × 105 conidia mL−1 on all adaxial leaf surfaces of plants at 60 days after emergence (growth stage 65. The fourth and fifth leaves of each plant were used to evaluate blast severity at 24, 36, 48, 72 and 96 h after inoculation (hai. At 96 hai, leaves were collected from plants to determine the foliar Si concentration. For cytological observations, leaf samples were randomly collected from the fourth and fifth leaves of each plant at 72 hai. The foliar Si concentration was higher in +Si plants (36 g kg−1 in comparison to -Si plants (2.6 g kg−1. This increased Si concentration was correlated with reduced fungal growth inside the epidermal cells and the development of blast symptoms on leaves. Strong fluorescence, which is an indication of the presence of flavonoids, was detected in the leaf cells of +Si plants using Neu’s and Wilson's reagents. A novel item of evidence is that, at the histochemical level, Si is involved in the potentiation of the biosynthetic pathway of flavonoids that increases wheat resistance to blast.

  5. Estudo histológico e histoquímico da glândula de Duvernoy de Clelia plumbea (Wied (Serpentes, Colubridae, Xenodontinae Histological and histochemical study of Duvernoy's gland from Clelia plumbea (Wied (Serpentes, Colubridae, Xenodontinae

    Directory of Open Access Journals (Sweden)

    Márcia Ferret Renner

    2000-09-01

    Full Text Available There are report two human envenenomations for species from Clelia Fitzinger, 1826 (opisthoglyph snake. The patients exhibited symptoms similar from bothropic accident. Then this work have with objective, the hystological and histochemical study of Duvernoy's gland from Clelia plumbea. For the hystological observation of the Duvernoy's gland were using, with paraffin's cutting edge, two methods: Hematoxylin + Eosin and Toluidine's Blue. With historesine's cutting edge were using three methods: Metilen's Blue, Toluidine's Blue + Floxine and Toluidine's Blue. For the histochemical detection were using six techniques, with paraffin's cutting edge: PAS, PAS + Alcian Blue pH 2,5, Alcian Blue pH 2,5, PAS + Salivar Amilase, Method of Lars Grimelius and Method of Lillie. It was evidenced the predominance of serous cells on Duvernoy's gland. In relation to the histochemical constituion of the duvernoy's gland, was verified the presence of glucoconjugates neuters, enzymes and cells with reduetive activity. By fact from this species to kill his preys by constriction, the predominance of serous cells in the duvernoy's gland, don' t must be direct related with the subjugation of his preys, but with the digestive processes. Histological and histochemical analysis showed cells of Duvernoy's gland are constituted for two populations: serous cells (prodution of enzymes and mucous cells (prodution of glucoconjugates.

  6. Cartilage canals in newborn dogs: histochemical and immunohistochemical findings

    Directory of Open Access Journals (Sweden)

    A. Di Giancamillo

    2016-09-01

    Full Text Available Cartilage canals (CCs are microscopic structures involved in secondary ossification centers (SOCs development. The features of CCs were investigated in the humeral and femoral proximal epiphyses of small-sized newborn dogs (from premature to 28 days after birth with histochemical and immunohistochemical approaches. Masson’s Trichrome revealed a ring-shaped area around CCs, which changes in colour from green (immature collagen to red (mature collagen as ossification progresses; perichondrium staining always matched the ring colour. Safranin-O was always negative. Immunohistochemical analysis revealed immunopositivity for both collagen type I and V around the CCs; collagen type II was negative. CCs count showed a tendency to be higher in the humerus than in the femur. This work enlightened for the first time changes in composition of CCs surrounding matrix during SOCs development in dogs, paving the way to further investigations.

  7. Histochemical discrimination of fibers in regenerating rat infraorbital nerve

    Science.gov (United States)

    Wilke, R. A.; Riley, D. A.; Sanger, J. R.

    1992-01-01

    In rat dorsal root ganglia, histochemical staining of carbonic anhydrase (CA) and cholinesterase (CE) yields a reciprocal pattern of activity: Sensory processes are CA positive and CE negative, whereas motor processes are CA negative and CE positive. In rat infraorbital nerve (a sensory peripheral nerve), we saw extensive CA staining of nearly 100% of the myelinated axons. Although CE reactivity in myelinated axons was extremely rare, we did observe CE staining of unmyelinated autonomic fibers. Four weeks after transection of infraorbital nerves, CA-stained longitudinal sections of the proximal stump demonstrated 3 distinct morphological zones. A fraction of the viable axons retained CA activity to within 2 mm of the distal extent of the stump, and the stain is capable of resolving growth sprouts being regenerated from these fibers. Staining of unmyelinated autonomic fibers in serial sections shows that CE activity was not retained as far distally as is the CA sensory staining.

  8. A futile cycle, formed between two ATP-dependant -glutamyl cycle enzymes, -glutamyl cysteine synthetase and 5-oxoprolinase: the cause of cellular ATP depletion in nephrotic cystinosis?

    Indian Academy of Sciences (India)

    Akhilesh Kumar; Anand Kumar Bachhawat

    2010-03-01

    Cystinosis, an inherited disease caused by a defect in the lysosomal cystine transporter (CTNS), is characterized by renal proximal tubular dysfunction. Adenosine triphosphate (ATP) depletion appears to be a key event in the pathophysiology of the disease, even though the manner in which ATP depletion occurs is still a puzzle. We present a model that explains how a futile cycle that is generated between two ATP-utilizing enzymes of the -glutamyl cycle leads to ATP depletion. The enzyme -glutamyl cysteine synthetase (-GCS), in the absence of cysteine, forms 5-oxoproline (instead of the normal substrate, -glutamyl cysteine) and the 5-oxoproline is converted into glutamate by the ATP-dependant enzyme, 5-oxoprolinase. Thus, in cysteine-limiting conditions, glutamate is cycled back into glutamate via 5-oxoproline at the cost of two ATP molecules without production of glutathione and is the cause of the decreased levels of glutathione synthesis, as well as the ATP depletion observed in these cells. The model is also compatible with the differences seen in the human patients and the mouse model of cystinosis, where renal failure is not observed.

  9. Mutation in the key enzyme of sialic acid biosynthesis causes severe glomerular proteinuria and is rescued by N-acetylmannosamine.

    Science.gov (United States)

    Galeano, Belinda; Klootwijk, Riko; Manoli, Irini; Sun, MaoSen; Ciccone, Carla; Darvish, Daniel; Starost, Matthew F; Zerfas, Patricia M; Hoffmann, Victoria J; Hoogstraten-Miller, Shelley; Krasnewich, Donna M; Gahl, William A; Huizing, Marjan

    2007-06-01

    Mutations in the key enzyme of sialic acid biosynthesis, uridine diphospho-N-acetylglucosamine 2-epimerase/N-acetylmannosamine (ManNAc) kinase (GNE/MNK), result in hereditary inclusion body myopathy (HIBM), an adult-onset, progressive neuromuscular disorder. We created knockin mice harboring the M712T Gne/Mnk mutation. Homozygous mutant (Gne(M712T/M712T)) mice did not survive beyond P3. At P2, significantly decreased Gne-epimerase activity was observed in Gne(M712T/M712T) muscle, but no myopathic features were apparent. Rather, homozygous mutant mice had glomerular hematuria, proteinuria, and podocytopathy. Renal findings included segmental splitting of the glomerular basement membrane, effacement of podocyte foot processes, and reduced sialylation of the major podocyte sialoprotein, podocalyxin. ManNAc administration yielded survival beyond P3 in 43% of the Gne(M712T/M712T) pups. Survivors exhibited improved renal histology, increased sialylation of podocalyxin, and increased Gne/Mnk protein expression and Gne-epimerase activities. These findings establish this Gne(M712T/M712T) knockin mouse as what we believe to be the first genetic model of podocyte injury and segmental glomerular basement membrane splitting due to hyposialylation. The results also support evaluation of ManNAc as a treatment not only for HIBM but also for renal disorders involving proteinuria and hematuria due to podocytopathy and/or segmental splitting of the glomerular basement membrane.

  10. Ca2+ cytochemical changes of hepatotoxicity caused by halothane and sevoflurane in enzyme-induced hypoxic rats

    Institute of Scientific and Technical Information of China (English)

    Wei-Feng Yu; Li-Qun Yang; Mai-Tao Zhou; Zhi-Qiang Liu; Quan Li

    2005-01-01

    AIM: To investigat the relation between hepatotoxicity of halothane and sevoflurane and altered hepatic calcium homeostasis in enzyme-induced hypoxic rats.METHODS: Forty-eight rats were pretreated with phenobarbital and randomly divided into six groups (eight in each group) and exposed to O2/N2/1.2 MAC anesthetics for :1 h: normal control (NC), 21% O2/79% N2; hypoxic control (HC), 14% O2/86%N2; normal sevoflurane (NS),21% O2/ N2/1.2MAC sevoflurane; hypoxic sevoflurane (HS), 14% O2/N2/1.2MAC sevoflurane; normal halothane (NH)21%O2/7g%N2/1.2MAC halothane; hypoxic halothane (HH), 14% O2/N2/1.2MAC halothane. Liver specimens and blood were taken 24 h after exposure to calcium and determined by EDX microanalysis.RESULTS: The liver of all rats given halothane (14%O2)had extensive centrilobular necrosis and denaturation.Morphologic damage was accompanied with an increase in serum glutamic pyruvic transminase. In groups NH and HH, more calcium was precipitated in cytoplasm and mitochondria.CONCLUSION: These results suggest that halothane increases cytosolic Ca2+ concentration in hepatocytes.Elevation in Ca2+ concentration is implicated in the mechanism of halothane-induced hepatotoxicity.sevoflurane is less effective in affecting hepatic calcium homeostasis than halothane.

  11. Altered cardiac bradykinin metabolism in experimental diabetes caused by the variations of angiotensin-converting enzyme and other peptidases.

    Science.gov (United States)

    Adam, Albert; Leclair, Patrick; Montpas, Nicolas; Koumbadinga, Gérémy Abdull; Bachelard, Hélène; Marceau, François

    2010-04-01

    The peptidases angiotensin-converting enzyme (ACE) and neutral endopeptidase 24.11 (NEP) mediate most of the kinin catabolism in normal cardiac tissue and are the molecular targets of inhibitory drugs that favorably influence diabetic complications. We studied the variations of those kininases in the myocardium of rats in experimental diabetes. ACE and NEP activities were significantly decreased in heart membranes 4-8weeks post-streptozotocin (STZ) injection. However, insulin-dependent diabetes did not modify significantly bradykinin (BK) half-life (t(1/2)) while the effect of both ACE (enalaprilat) and ACE and NEP (omapatrilat) inhibitors on BK degradation progressively decreased, which may be explained by the upregulation of other unidentified metallopeptidase(s). In vivo insulin treatment restored the activities of both ACE and NEP. ACE and NEP activities were significantly higher in hearts of young Zucker rats than in those of Sprague-Dawley rats. BK t(1/2) and the effects of peptidase inhibitors on t(1/2) varied accordingly. It is concluded that kininase activities are subjected to large and opposite variations in rat cardiac tissue in type I and II diabetes models. A number of tissue or molecular factors may determine these variations, such as remodeling of cardiac tissue, ectoenzyme shedding to the extracellular fluid and the pathologic regulation of peptidase gene expression.

  12. Mutations in Biosynthetic Enzymes for the Protein Linker Region of Chondroitin/Dermatan/Heparan Sulfate Cause Skeletal and Skin Dysplasias

    Directory of Open Access Journals (Sweden)

    Shuji Mizumoto

    2015-01-01

    Full Text Available Glycosaminoglycans, including chondroitin, dermatan, and heparan sulfate, have various roles in a wide range of biological events such as cell signaling, cell proliferation, tissue morphogenesis, and interactions with various growth factors. Their polysaccharides covalently attach to the serine residues on specific core proteins through the common linker region tetrasaccharide, -xylose-galactose-galactose-glucuronic acid, which is produced through the stepwise addition of respective monosaccharides by four distinct glycosyltransferases. Mutations in the human genes encoding the glycosyltransferases responsible for the biosynthesis of the linker region tetrasaccharide cause a number of genetic disorders, called glycosaminoglycan linkeropathies, including Desbuquois dysplasia type 2, spondyloepimetaphyseal dysplasia, Ehlers-Danlos syndrome, and Larsen syndrome. This review focused on recent studies on genetic diseases caused by defects in the biosynthesis of the common linker region tetrasaccharide.

  13. Creatine and pyruvate prevent the alterations caused by tyrosine on parameters of oxidative stress and enzyme activities of phosphoryltransfer network in cerebral cortex of Wistar rats.

    Science.gov (United States)

    de Andrade, Rodrigo Binkowski; Gemelli, Tanise; Rojas, Denise Bertin; Bonorino, Narielle Ferner; Costa, Bruna May Lopes; Funchal, Cláudia; Dutra-Filho, Carlos Severo; Wannmacher, Clovis Milton Duval

    2015-01-01

    Tyrosine accumulates in inborn errors of tyrosine catabolism, especially in tyrosinemia type II. In this disease caused by tyrosine aminotransferase deficiency, eyes, skin, and central nervous system disturbances are found. In the present study, we investigated the chronic effect of tyrosine methyl ester (TME) and/or creatine plus pyruvate on some parameters of oxidative stress and enzyme activities of phosphoryltransfer network in cerebral cortex homogenates of 21-day-old Wistar. Chronic administration of TME induced oxidative stress and altered the activities of adenylate kinase and mitochondrial and cytosolic creatine kinase. Total sulfhydryls content, GSH content, and GPx activity were significantly diminished, while DCFH oxidation, TBARS content, and SOD activity were significantly enhanced by TME. On the other hand, TME administration decreased the activity of CK from cytosolic and mitochondrial fractions but enhanced AK activity. In contrast, TME did not affect the carbonyl content and PK activity in cerebral cortex of rats. Co-administration of creatine plus pyruvate was effective in the prevention of alterations provoked by TME administration on the oxidative stress and the enzymes of phosphoryltransfer network, except in mitochondrial CK, AK, and SOD activities. These results indicate that chronic administration of TME may stimulate oxidative stress and alter the enzymes of phosphoryltransfer network in cerebral cortex of rats. In case this also occurs in the patients affected by these disorders, it may contribute, along with other mechanisms, to the neurological dysfunction of hypertyrosinemias, and creatine and pyruvate supplementation could be beneficial to the patients.

  14. Droplet Microfluidics Platform for Highly Sensitive and Quantitative Detection of Malaria-Causing Plasmodium Parasites Based on Enzyme Activity Measurement

    DEFF Research Database (Denmark)

    Juul, Sissel; Nielsen, Christine Juul Fælled; Labouriau, Rodrigo

    2012-01-01

    detectable at the single-molecule level. Combined with a droplet microfluidics lab-on-a-chip platform, this design allowed for sensitive, specific, and quantitative detection of all human-malaria-causing Plasmodium species in single drops of unprocessed blood with a detection limit of less than one parasite...... detection of even a few parasites is becoming increasingly important for the continued combat against the disease. We believe that the presented droplet microfluidics platform, which has a high potential for adaptation to point-of-care setups suitable for low-resource settings, may contribute significantly...

  15. Histological and histochemical changes in placenta of diabetic pregnant females and its comparision with normal placenta

    Directory of Open Access Journals (Sweden)

    Vineeta Tewari

    2011-03-01

    Full Text Available Objective: To investigate the histological and histochemical changes in placenta of diabetic pregnant females and compare them with normal placenta. Methods: The histological and histochemical features of 60 placenta, 30 obtained from normal pregnant females and 30 from diabetic pregnant females, were studied. These placenta were obtained from Department of Obstetrics and Gynaecology, GSVM Medical College Kanpur and ERA ’s Lucknow Medical College and Hospital Lucknow. Results: On histological examination, the diabetic placenta showed increased syncytial knots, fibrinoid necrosis, trophoblastic basement membrane thickening, villous stromal fibrosis, villous oedema, crowding of villi, thickening of vessel wall and fibrin deposition. On histochemical study it was found that the PAS reactivity was stronger in diabetic placenta as compared to normal. Sudan Black reactivity was higher among diabetic placenta in comparison to normal placenta. Conclusions: It is concluded that distinct histological and histochemical changes could be seen in placenta of diabetic pregnant females.

  16. Evaluation of the gene encoding the enzyme βHPMEH for the bacterial wilt inhibition caused by Ralstonia solanacearum

    Directory of Open Access Journals (Sweden)

    Elizabeth Fernandez

    2015-10-01

    Full Text Available Ralstonia solanacearum is the causal agent of the devastating bacterial wilt disease that attacks important agricultural crops such as potato, tomato, banana, among others, causing serious yield losses. Control of R. solanacearum is difficult because of its wide range of alternate hosts, its long survival in soil, its biological and genetic variation, the lack of natural resistance sources and the insufficiency of the appropriate chemical control measures. Quorum sensing is the term that describes the phenomenon whereby the accumulation of molecules allows bacteria to know the number of bacteria found in the environment (population density. R. solanacearum has a quorum sensing system for the regulation of the expression of virulence genes; the molecule 3-OH-PAME is the self-regulatory signal. The molecule ΒHPMEH hydrolyzes 3-OH-PAME nullifying the signal of virulence, and thus, the quorum sensing communication in R. solanacearum. In order to evaluate the βhpmeh gene we designed two vectors that express this gene under the control of two different promoters. Both vectors were verified by restriction analysis and sequencing. Agroinfiltration assays were used to analyze gene expression and the effect against R. solanacearum in potato (Solanum tuberosum leaves. The results of the transient expression experiments showed that the expression of gene βhpmeh caused a delay in the appearance of symptoms of bacterial wilt and thus is a good candidate for whole genetic plant transformation.

  17. Hairy cell leukemia: enzyme-histochemical and ultrastructural investigation of one case.

    Science.gov (United States)

    Pilotti, S; Carbone, A; Lombardi, L; Tavolato, C; Rilke, F

    1978-10-31

    The investigation was carried out on blood smears, bone marrow aspirates, one lymph node biopsy, and the surgically removed spleen of a 53-year-old man with hairy cell leukemia. In the blood smears stained with May-Grünwald-Giemsa, 60 to 70% of the hairy cells contained tubular inclusions that corresponded to the ribosome-lamella complexes demonstrated at electron microscopy. In blood smears, imprints and cryostatic sections of the lymph node and of the spleen, hairy cells revealed tartrate-resistant acid phosphatase, beta-glucuronidase and adenosine-triphosphatase activity. In the spleen neutral esterase and alkaline phosphatase demonstrated the numerical increase of the histiocytes, which ultrastructurally displayed phagocytic activity. The presence in the spleen of pseudosinuses lined by hairy cells was confirmed by electron microscopy as well as by cytoenzymology.

  18. [Immuno-histochemical study of medullary thyroid carcinoma].

    Science.gov (United States)

    Bessho, T

    1987-07-01

    Five peptide hormones including calcitonin (CT) and gastrin-releasing peptide (GRP), serotonin (5HT), CEA, nervous tissue specific proteins and monoclonal antibody Leu-7 were immuno-histochemically studied on 60 cases of medullary thyroid carcinoma (MTC). In addition, localization of varied products in the tumor cells and its relations with the clinical features in some cases were evaluated. MTC contains a variety of products in many cases, and CT and CEA were positive in all cases. In 50 of the 57 cases (87.7%), GRP was positive, which suggested that GRP could be a novel tumor marker for this tumor. Furthermore, in tumor cells and C-cell hyperplastic foci, identical cells were sometimes revealed to possess both CT and GRP. Existence of somatostatin (SS), substance-P (SP), beta-MSH, 5 HT, Leu-7 and NSE in the tumor cells were confirmed. NSE was positive in 32 of the 47 cases (61.8%) which could confirm that MTC possesses neuroendocrine nature. In two cases of autopsy in which the tumors were highly malignant in clinical course and undifferentiated in histology, most tumor cells showed poor stainability for peptide hormones, suggesting that specific qualities as neuroendocrine tumor had been lost. In familial cases, the tumor tended to contain multiple substances.

  19. Histochemical and Immunohistochemical Study of Peripolar Cells in Sheep

    Directory of Open Access Journals (Sweden)

    Passos Joana

    2013-01-01

    Full Text Available Peripolar cells are granulated cells located in the vascular pole of the renal corpuscle. Even though these cells have already been described, there are still many unknown histological and physiological characteristics. We carried out histochemical and immunohistochemical analyses of peripolar cells in sheep and compared their number in both normal and injured kidneys, discriminating according to the age of the animal. We tested HE, Toluidine Blue, PAS, and Masson's Trichrome stains to select the best stain for identification and quantification. Masson Trichrome yielded the best results and was selected for this purpose. We identified the cells by the presence of cytoplasmatic granules and by their position in the vascular pole. We found no statistically significant association between the number of peripolar cells and the age of the animal or the occurrence of lesions. In the immunohistochemical analysis, we found that the cells were positive to α-smooth muscle actin and less consistently positive to NSE and S100 protein. Chromogranin A, cyclooxygenase-2, AE1/AE3, and Wide Spectrum Cytokeratin and desmin yielded negative results. We conclude that although there was evidence of a contractile function, there was no evidence to support that peripolar cells have either a neuroendocrine or an epithelial nature.

  20. Use of enzymes to minimize the rheological dough problems caused by high levels of damaged starch in starch-gluten systems.

    Science.gov (United States)

    Barrera, Gabriela N; León, Alberto E; Ribotta, Pablo D

    2016-05-01

    During wheat milling, starch granules can experience mechanical damage, producing damaged starch. High levels of damaged starch modify the physicochemical properties of wheat flour, negatively affecting the dough behavior as well as the flour quality and cookie and bread making quality. The aim of this work was to evaluate the effect of α-amylase, maltogenic amylase and amyloglucosidase on dough rheology in order to propose alternatives to reduce the issues related to high levels of damaged starch. The dough with a high level of damaged starch became more viscous and resistant to deformations as well as less elastic and extensible. The soluble fraction of the doughs influenced the rheological behavior of the systems. The α-amylase and amyloglucosidase reduced the negative effects of high damaged starch contents, improving the dough rheological properties modified by damaged starch. The rheological behavior of dough with the higher damaged-starch content was related to a more open gluten network arrangement as a result of the large size of the swollen damaged starch granules. We can conclude that the dough rheological properties of systems with high damaged starch content changed positively as a result of enzyme action, particularly α-amylase and amyloglucosidase additions, allowing the use of these amylases and mixtures of them as corrective additives. Little information was reported about amyloglucosidase activity alone or combined with α-amylase. The combinations of these two enzymes are promising to minimize the negative effects caused by high levels of damaged starch on product quality. More research needs to be done on bread quality combining these two enzymes. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  1. The Use of Recently Developed Histochemical Markers for Localizing Neurotoxicant Induced Regional Brain Pathologies

    Directory of Open Access Journals (Sweden)

    Sumit Sarkar

    2014-04-01

    Full Text Available Neuronal and vascular brain components are interrelated morphologically, physiologically and developmentally. Due to this close interrelationship, it is often difficult to understand the cause and effect relationship between neuronal vs. vascular dysfunction and pathology. This review will discuss four of the more promising recent developments for detecting vascular pathology, and will compare them with the labeling pattern seen with markers of glial and neuronal pathology; following exposure to well characterized neurotoxicants. To detect the vascular dysfunction in the brain, we recently developed a Fluoro-Turquoise gelatin conjugate (FT-gel, a fluorescent probe that helps to delineate between healthy vs. sclerotic vessels. Similarly, we have investigated the potential for Fluoro-Gold to label in vivo all the endothelial cells in the brain as they co-localize with RECA, an endothelial cell marker. We have also developed Amylo-Glo, a fluorescent tracer that can detect neurotoxic A-beta aggregates in the brain. In this article, we will discuss the potential use of these novel histochemical markers to study the neurotoxicant induced brain. We will also discuss neurovascular strategies that may offer novel therapeutic opportunities for neurodegenerative disorders.

  2. Optimization of the tartrate-resistant acid phosphatase detection by histochemical method

    Science.gov (United States)

    Galvão, M.J.; Santos, A. R.; Ribeiro, M.D.; Ferreira, A.; Nolasco, F.

    2011-01-01

    According to the new kidney disease improving global outcomes (KDIGO) guidelines, the term of renal osteodystrophy, should be used exclusively in reference to the invasive diagnosis of bone abnormalities. Due to the low sensitivity and specificity of biochemical serum markers of bone remodelling, the performance of bone biopsies is highly stimulated in dialysis patients and after kidney transplantation. The tartrate-resistant acid phosphatase (TRACP) is an iso-enzyme of the group of acid phosphatases, which is highly expressed by activated osteoclasts and macrophages. TRACP in osteoclasts is in intracytoplasmic vesicles that transport the products of bone matrix degradation. Being present in activated osteoclasts, the identification of this enzyme by histochemistry in undecalcified bone biopsies is an excellent method to quantify the resorption of bone. Since it is an enzymatic histochemical method for a thermolabile enzyme, the temperature at which it is performed is particularly relevant. This study aimed to determine the optimal temperature for identification of TRACP in activated osteoclasts in undecalcified bone biopsies embedded in methylmethacrylate. We selected 10 cases of undecalcified bone biopsies from hemodialysis patients with the diagnosis of secondary hyperparathyroidism. Sections of 5 µm were stained to identify TRACP at different incubation temperatures (37°, 45°, 60°, 70° and 80°C) for 30 minutes. Activated osteoclasts stained red and trabecular bone (mineralized bone) was contrasted with toluidine blue. This approach also increased the visibility of the trabecular bone resorption areas (Howship lacunae). Unlike what is suggested in the literature and in several international protocols, we found that the best results were obtained with temperatures between 60°C and 70°C. For technical reasons and according to the results of the present study, we recommended that, for an incubation time of 30 min, the reaction should be carried out at 60

  3. Lectin Histochemical Study of Vasculogenesis During Rat Pituitary Morphogenesis

    Directory of Open Access Journals (Sweden)

    Ali Reza Ebrahimzadeh Bideskan

    2011-01-01

    Full Text Available Objective(s The aim of this study was to investigate glycoconjugates distribution patterns as well as their changes during the course of pituitary portal vasculogenesis and angiogenesis.Materials and MethodsFormalin fixed paraffin sections of 10 to 20 days of Sprague Dawly rat fetuses were processed for histochemical studies using four different horseradish peroxidase (HRP conjugated lectins. Orange peel fungus (OFA, Vicica villosa (VVA, Glycine max (SBA and Wistaria floribunda (WFA specific for α-L-Fucose, D-Gal, α, ß-D-GalNAc and D- GalNAc terminal sugars of glycoconjugates respectively.ResultsOur finding indicated that adenohypophysal cells reacted with OFA on gestational day 10 (E10 and increased progressively to E14. Staining intensity did not change from days 14 to17, then after increased following days to E20 significantly (P< 0.05. A few cells around Rathke’s pouch reacted with VVA on E13, increased to E14 and decreased significantly afterward (P< 0.05. Reaction of some cells around Rathke’s pouch reacted with SBA on E14. This visible reaction was the same as E18 and decreased later (P< 0.05. Many cells around Rathke’s pouch reacted with WFA on E13 and increased on E 14 and E15 and decreased thereafter (P< 0.05.ConclusionReactions of OFA and other tested lectins with endothelial cells around Rathke’s pouch and developing pars distalis were different. These results suggest that embryonic origin of hypophiseal pituitary portal (HPP system endothelial cells are not the same and our finding also indicated that glycoconjugates with terminal sugars α-L-Fucose, D-Gal, α, ß-D-GalNAc may play critical role(s in cell interactions and tissue differentiations such as vasculogensis and angiogenesis as well as other developmental precursors in formation of the pituitary gland.

  4. Histological and histochemical characterisation of the equine soft palate muscles.

    Science.gov (United States)

    Hawkes, C S; Hahn, C N; Dixon, P M

    2010-07-01

    Dysfunction of the muscles is implicated in the pathogenesis of intermittent dorsal displacement of the soft palate (DDSP) in exercising horses. The histological features of normal equine soft palate muscles have not been previously described. To describe the histological and morphometric features of normal equine soft palate muscles. The palatinus, palatopharyngeus, levator veli palatini and tensor veli palatini muscles of 6 Thoroughbred type horses were examined histologically and histochemically to assess their general morphology, fibre-type distribution and mean fibre diameter. The muscles of all 6 specimens showed very similar characteristics, including a low proportion of type 1 muscle fibres in the 4 different muscles examined, with the 3 muscles innervated by the pharyngeal branch of the vagus (i.e. palatopharyngeus, palatinus and levator veli palatini) having significantly fewer (Pmuscle fibres compared to the tensor veli palatini. The mean fibre diameters were significantly (Pmuscles except the palatinus. Considerable variability in mean fibre size diameter was evident in all muscles examined, with type 1 fibres in the levator veli palatini and rostral fasciculus of the palatopharyngeus and both fibre types in the caudal fasciculus of the palatopharyngeus having the most marked variation, with coefficients of variation >250 and the latter also having high levels of endomysial connective tissue. The palatinus had least variation in fibre size. Morphological characterisation of the normal soft palate muscles may provide reference values for future comparative studies with samples obtained from horses with palatal dysfunction. The palatinus appears to be the best muscle to histologically examine for evidence of muscle abnormality.

  5. Histological and histochemical characterization on stomach of Mystus cavasius (Hamilton, Oreochromis niloticus (Linnaeus and Gudusia chapra (Hamilton: Comparative study

    Directory of Open Access Journals (Sweden)

    Saroj K. Ghosh

    2015-05-01

    Full Text Available The histological features and histochemical characterization of the stomach were investigated in Mystus cavasius (Hamilton, Oreochromis niloticus (Linnaeus and Gudusia chapra (Hamilton having different feeding habits. Histologically the stomach of all the three fishes was made up of mucosa, submucosa, muscularis and serosa. The mucosa of superficial epithelium consists of a single layer of compactly arranged columnar epithelial cells. Prominent gastric glands are present in M. cavasius whereas in G. chapra the gastric glands are totally absent in the gizzard like stomach. However, in O. niloticus tubular gastric glands are present in the glandular epithelium of caecal like stomach. The distribution and chemical nature of mucopolysaccharides in the aforementioned fishes were studied histochemically by employing Periodic Acid Schiff’s in combination with the Alcian Blue (PAS–AB technique. Columnar epithelial cells lining the mucosa of the stomach including mucosal border were provided with exclusively neutral mucin which was probably involved in the protective functions against acid and enzymes. The different intensities of reaction of Best Carmine (BC for glycogen in the epithelial lining and gastric glands of the stomach of the aforesaid three fish species under study were discussed. The intense reaction for protein and tryptophan was noticed in the gastric epithelium and gastric glands of M. cavasius probably due to accumulation of zymogen granules in the gastric glands. On the contrary, moderate reaction for protein and tryptophan was associated with the epithelial cells and gastric glands of O. niloticus and G. chapra. The cytoarchitecture and different degrees of localization of mucopolysaccharides, glycogen, protein and tryptophan in the stomach of M. cavasius, O. niloticus and G. chapra were correlated with the functional significance of the region concerned.

  6. Bile acid malabsorption or disturbed intestinal permeability in patients treated with enzyme substitution for exocrine pancreatic insufficiency is not caused by bacterial overgrowth

    DEFF Research Database (Denmark)

    Madsen, Jan Lysgård; Graff, Jesper; Philipsen, Else Kirstine;

    2003-01-01

    INTRODUCTION: In some patients with severe exocrine pancreatic insufficiency, enzyme replacement therapy will not lead to clinical improvement or reduction of steatorrhea. Therefore, other mechanisms separately or in interplay with reduced enzyme secretion might be responsible for malabsorption...

  7. Bile acid malabsorption or disturbed intestinal permeability in patients treated with enzyme substitution for exocrine pancreatic insufficiency is not caused by bacterial overgrowth

    DEFF Research Database (Denmark)

    Madsen, Jan Lysgård; Graff, Jesper; Philipsen, Else Kirstine

    2003-01-01

    INTRODUCTION: In some patients with severe exocrine pancreatic insufficiency, enzyme replacement therapy will not lead to clinical improvement or reduction of steatorrhea. Therefore, other mechanisms separately or in interplay with reduced enzyme secretion might be responsible for malabsorption...

  8. Use of Peptide-Based Enzyme-Linked Immunosorbent Assay followed by Immunofluorescence Assay To Document Ehrlichia chaffeensis as a Cause of Febrile Illness in Nicaragua.

    Science.gov (United States)

    Chikeka, Ijeuru; Matute, Armando J; Dumler, J Stephen; Woods, Christopher W; Mayorga, Orlando; Reller, Megan E

    2016-06-01

    Ehrlichia chaffeensis, the etiologic agent of human monocytic ehrlichiosis (HME), has been extensively studied as a cause of acute febrile illness and an emerging tick-borne zoonosis in the United States. Limited data suggest its presence in other regions, including Central and South America but not Nicaragua to date. Diagnosis of E. chaffeensis infection by indirect immunofluorescence assay (IFA) is the reference standard due to its presumed high sensitivity and specificity, but IFA is impractical, variably reproducible, and cumbersome for large epidemiologic studies and for clinical diagnosis in resource-poor regions. We evaluated a high-throughput, objective peptide-based enzyme-linked immunosorbent assay (ELISA) for use alone or in combination with IFA. We found that it performed best as a screening test (sensitivity, 100%; specificity, 84%) to reduce the proportion of serum samples that were required by the more cumbersome and subjective IFA testing to ELISA is positive), we identified E. chaffeensis or a serologically and antigenically similar organism as a heretofore unrecognized cause of acute febrile illness in humans in Nicaragua and demonstrated the utility of the peptide ELISA as a screening tool for large-scale clinical studies.

  9. Effect of pollution on some freshwater species. I. histochemical and biochemical features of lead pollution on some organs of Viviparus viviparus L. (Mollusca Gastropoda)

    Energy Technology Data Exchange (ETDEWEB)

    Benedetti, I.; Benedetti, L.; Bolognani, L.; Bolognani Fantin, A.M.; Marini, M.; Ottaviani, E.

    1982-01-01

    Experimental lead pollution was studied in some organs (foot, mantle and digestive gland) of Viviparus viviparus L. The amount of lead contained after 48h, 96 h and one week of pollution were established using an atomic absorption spectrophotometer. On the basis of physicochemical determination, it turns out that lead is mainly concentrated in the mantle. The biochemical tests (cholesterol, sulpholipids and phospholipids) were aimed at evaluating the lipids involved in the membranes. The histochemical research was carried out chiefly to evaluate the modifications of polysaccharides and proteins. Some hydrolytic enzymes (Na+ and K+ dependent ATPase) and some ooreductive enzymes (NADH+ and NADPH+ dependent diaphorases, D-lactate dehydrogenase, succinate dehydrogenase and glucose-6-P-dehydrogenase) were also tested. The digestive gland is the most severely damaged organ as proved by histomorphological and biochemical analyses.

  10. Thiamethoxam causes histochemical changes in the liver of Aristichthys nobilis Rich., 1845

    Directory of Open Access Journals (Sweden)

    STELA STOYANOVA

    2015-12-01

    Full Text Available In the present study, we aimed to investigate the effects of the neonicotinoid insecticide thiamethoxam on the hepatic glycogen in bighead carp (Aristichthys nobilis Rich.. Fish were exposed to 6.6 mg/L, 10 mg/L and 20 mg/L of the insecticide under laboratory conditions for 96 hours. The PAS-reaction was applied to liver cryostat sections in order to indicate the amount of glycogen. The results showed that the hepatic glycogen amount increased with increasing the insecticide concentrations. On the other hand, we observed glycogen conglomerates in certain hepatocytes. Hence, our results demonstrated an enhanced process of glyconeogenesis in the fish liver under the influence of thiamethoxam.

  11. The porcine ear skin as a model system for the human integument: influence of storage conditions on basic features of epidermis structure and function--a histological and histochemical study.

    Science.gov (United States)

    Meyer, W; Zschemisch, N H; Godynicki, S

    2003-01-01

    Based on careful tissue processing, detailed structural analysis, and histochemical as well as cytophotometrical evaluation of the epidermis, the study presents data with respect to changes of tissue integrity during two storing modes (room temperature and 4 degrees C) and various storage times of the porcine auricle. Structural degeneration was first noted in the barrier region of the epidermis from where such changes spread, independent of storage conditions, from small horizontal necrotic islands and continuously with increasing storage time. The histochemical results corroborated these observations, emphasizing, however, that the lower epidermal layers seemed intact for a longer time period than the upper layers. Cytophotometrical evaluation of histochemical stainings showed, with regard to the enzyme succinate dehydrogenase, that oxidative metabolism was negatively affected in the early stages of storage, whereas epidermal lipids (neutral fats, glycolipids) remained relatively stable, even during storage at room temperature. In conclusion, it was obvious that the barrier region is the most sensitive element of the porcine ear epidermis. Taking into consideration that this part of the epidermis is most important for permeation studies, it seems reasonable to avoid any storage of porcine auricles at room temperature, and to use only auricles that have been stored at 4 degrees C for not more than 4 to 6 hours, immediately after delivery from the slaughter-house. In this way better tissue preservation can be achieved, whereby the use of shinkage-free water-soluble plastic embedding would generally improve the histological control of structural integrity, and the application of an easy to handle enzyme histochemical procedure (e.g. succinate dehydrogenase demonstration) to unfixed fresh-frozen sections would help to control basic aspects of tissue functions. The results are discussed in relation to the use of porcine integument as a model in human dermatological

  12. Somatic-cell selection is a major determinant of the blood-cell phenotype in heterozygotes for glucose-6-phosphate dehydrogenase mutations causing severe enzyme deficiency

    Energy Technology Data Exchange (ETDEWEB)

    Filosa, S.; Giacometti, N.; Wangwei, C.; Martini, G. [Istituto Internazionale di Genetica e Biofisica, Naples (Italy)] [and others

    1996-10-01

    X-chromosome inactivation in mammals is regarded as an essentially random process, but the resulting somatic-cell mosaicism creates the opportunity for cell selection. In most people with red-blood-cell glucose-6-phosphate dehydrogenase (G6PD) deficiency, the enzyme-deficient phenotype is only moderately expressed in nucleated cells. However, in a small subset of hemizygous males who suffer from chronic nonspherocytic hemolytic anemia, the underlying mutations (designated class I) cause more-severe G6PD deficiency, and this might provide an opportunity for selection in heterozygous females during development. In order to test this possibility we have analyzed four heterozygotes for class I G6PD mutations: two with G6PD Portici (1178G{r_arrow}A) and two with G6PD Bari (1187C{r_arrow}T). We found that in fractionated blood cell types (including erythroid, myeloid, and lymphoid cell lineages) there was a significant excess of G6PD-normal cells. The significant concordance that we have observed in the degree of imbalance in the different blood-cell lineages indicates that a selective mechanism is likely to operate at the level of pluripotent blood stem cells. Thus, it appears that severe G6PD deficiency affects adversely the proliferation or the survival of nucleated blood cells and that this phenotypic characteristic is critical during hematopoiesis. 65 refs., 6 figs., 3 tabs.

  13. Histochemical and immunohistochemical analysis of ruptured atherosclerotic abdominal aortic aneurysm wall

    Directory of Open Access Journals (Sweden)

    Tanasković Irena

    2010-01-01

    Full Text Available Background/Aim. The main complication of the atherosclerotic abdominal aortic aneurism (AAA is her rupture that begins with lesion in intima and rupture. The purpose of this work was to determine immunocytochemical and morphofunctional characteristics of the cells in aortic wall in ruptured atherosclerotic abdominal aortic aneurysm. Method. During the course of this study, 20 samples of atherosclerotic AAA were analyzed, all of them obtained during authopsy. The samples were fixed in 4% formalin and embedded in paraffin. Sections of 5 μm thickness were stained histochemically (of Heidenhain azan stain and Periodic acid Schiff - PAS stain and immunocytochemically using a DAKO LSAB+/HRP technique to identify α-smooth muscle actin (α-SMA, vimentin, myosin heavy chains (MHC, desmin, S-100 protein, CD45 and CD68 (DAKO specification. Results. The results of our study showed that ruptured atherosclerotic AAA is characterized by a complete absence of endothelial cells, the disruption of basal membrane and internal elastic lamina, as well as a presence of the remains of hypocellular complicated atherosclerotic lesion in intima. On the plaque margins, as well as in the media, smooth muscle cells (SMCs are present, which express a α-SMA and vimentin (but without MHC or desmin expression, as well as leukocyte infiltration, and a large number of foam cells. Some of the foam cells show a CD68-immunoreactivity, while the others show vimentin- and S-100 protein-immunoreactivity. Media is thinned out with a disorganized elastic lamellas, while adventitia is characterized by inflammatory inflitrate (infection. Conclusion. Rupture of aneurysm occurs from the primary intimal disruption, which spreads into thinned out media and adventitia. Rupture is caused by unstable atherom, hypocellularity, loss of contractile characteristics of smooth muscle cells in intima and media, neovascularization of the media, as well as by the activity of the macrophages in the

  14. Implementation of different histochemical methods in diagnostics of brain Aspergillosis in turkey chicks

    Directory of Open Access Journals (Sweden)

    Kureljušić Branislav

    2011-01-01

    Full Text Available Aspergillosis is a frequent fungal disease in different species of birds and mammals caused by fungi of the genus Aspergillus. It is characterized by inflammatory changes primarily in the respiratory system, even though it sometimes takes on a generalized form when several organ systems are affected. Mucotic-granulomatous meningoencephalitis with a predominant localization in the cerebellum has been described in turkeys, ducks and geese. Within this paper, examinations have been performed on a flock of broiler turkeys aged 12 days who had sustained evident neurological disorders in the form of ataxy, torticollis, paresis, and paralysis of the hind extremities and wings. In three of the ten autopsied chicks the macroscopic findings indicated granulomatous encephalitis of the cerebellum. A white-coloured granuloma, around 3mm in diameter, was situated cranioventrally and was clearly visible on the sagital section of the cerebellum. Mucological examinations of the cerebellum lesion resulted in the isolation of the fungi Aspergillus fumigatus. Haematoxylin-eosin (HE, Grocott and PAS methods were used for the evaluation of histopathological changes and proving Aspergillus fumigatusa hyphae. The microscopic examination of brain tissue sections stained with the HE method revealed the existence of a granuloma with a centrally placed necrotic area. The necrotic area was infiltrated with heterophilic granulocytes and surrounded by macrophage, giant cells and lymphocytes. A connective tissue capsule was located on the periphery of the granuloma. The fungi hyphae, as integral parts of the granuloma, were difficult to observe, and in some samples stained using the HE method they could not be seen at all. On the other hand, sections stanied using the Grocott and PAS methods showed prominent septed and branched hyphae in different parts of the granuloma. With the objective of making an etiological diagnosis of mucotic diseases, it is necessary to apply several

  15. Biochemical, Histological And Histochemical Studies On The Effect Of Sodium Barbital On The Renal Tissue Of Albino Mice

    Directory of Open Access Journals (Sweden)

    Samia M. Sakr*, Shadia Ali Radwan*, Aziza M. El Wessemy

    2008-03-01

    Full Text Available Introduction:Anaesthetic drugs are nowadays used on a large scale in surgical operations as well as in other various medical purposes. Sodium barbital is a derivative of barbituric acid and is widely used on short surgical operations and other various medication. However, such anaesthetic drug has been reported to evoke many serious alterations as a result of its application. Materials and Methods:The experimental animals (30 mice-weighing 25-30 g were divided into 3 groups (10/group, the first group served as a control group (i.e. injection with saline, while the other two groups were treated daily with the therapeutic dose of 60 mg/kg.b.wt sodium barbital (i.p. for 7 days (short-term group and 21 days (long-term group as repeated daily doses. Blood sera and kidney samples were collected for physiological, histological and histochemical studies. Results:The results obtained showed a significant increase in urea, blood urea nitrogen, uric acid and creatinine levels in all treated groups. On the other hand serum total protein and albumin levels showed a significant decrease in both treated groups, while the globulin showed a significant decrease only in the long term group. The applied dose of sodium barbital caused histophathological alterations in the renal tissue mainly in the cortex such as damage and shrinkage of the Malpighian corpuscles, cloudy swelling and necrosis of the cells of the proximal convoluted tubules. Also, distal convoluted tubules exhibited degenerated features. In the histochemical studies, polysaccharides were progressively reduced in both short and long-term groups, while the total proteins showed a reduction in the short term group and considerably increase in the long term group. Conclusion: So these results came to conclusion that barbiturates should be prohibited and carefully used specially when prescribed as tranquilizer

  16. Acquired melanosis caused by acorn ingestion in the Nero Siciliano pig.

    Science.gov (United States)

    Lanteri, G; Marino, F; Laganà, G; Bellocco, E; Barreca, D; Liotta, L; Sfacteria, A; Macrì, B

    2009-03-01

    In this study, an acquired pigmentation in Nero Siciliano pigs is reported and evaluated by a multidisciplinary approach to support the hypothesis it is caused by an ingested material. A total of 18 pigs were studied. Fourteen conventionally slaughtered animals showed black discoloration of lymph nodes. The lymph nodes were normal in size and shape but showed diffuse black discoloration of the cortex and medulla. Melanosis of fat was observed in 2 animals and was limited to the back. Histochemical tests performed on tissues enabled identification and differentiation of the pigment. Immunohistochemical staining for macrophage markers showed macrophages containing a variable amount of melanin-like granules. Stains for human melanoma, as well as S-100 protein, did not show any reaction. Histochemical methods for tyrosinase showed colorimetric patterns that confirmed the presence of the enzyme in acorns. The activity was mostly latent. A high tannin content was demonstrated, reaching about 76% of the total phenolic compounds. Our data, and the well-known steps on melanin formation, permit us to hypothesize that swine tyrosinase could act on phenolic substances found in acorns. Tyrosinase activation could take place in genetically predisposed swine after acorns are eaten, and this event could increase the biosynthesis and the anomalous storage of melanin.

  17. A comparison of tabun-inhibited rat brain acetylcholinesterase reactivation by three oximes (HI-6, obidoxime, and K048) in vivo detected by biochemical and histochemical techniques.

    Science.gov (United States)

    Bajgar, Jiri; Hajek, Petr; Zdarova, Jana Karasova; Kassa, Jiri; Paseka, Antonin; Slizova, Dasa; Krs, Otakar; Kuca, Kamil; Jun, Daniel; Fusek, Josef; Capek, Lukas

    2010-12-01

    Tabun belongs to the most toxic nerve agents. Its mechanism of action is based on acetylcholinesterase (AChE) inhibition at the peripheral and central nervous systems. Therapeutic countermeasures comprise administration of atropine with cholinesterase reactivators able to reactivate the inhibited enzyme. Reactivation of AChE is determined mostly biochemically without specification of different brain structures. Histochemical determination allows a fine search for different structures but is performed mostly without quantitative evaluation. In rats intoxicated with tabun and treated with a combination of atropine and HI-6, obidoxime, or new oxime K048, AChE activities in different brain structures were determined using biochemical and quantitative histochemical methods. Inhibition of AChE following untreated tabun intoxication was different in the various brain structures, having the highest degree in the frontal cortex and reticular formation and lowest in the basal ganglia and substantia nigra. Treatment resulted in an increase of AChE activity detected by both methods. The highest increase was observed in the frontal cortex. This reactivation was increased in the order HI-6 tabun, reactivation in various parts of the brain is not of the same physiological importance. AChE activity in the pontomedullar area and frontal cortex seems to be the most important for the therapeutic effect of the reactivators. HI-6 was not a good reactivator for the treatment of tabun intoxication.

  18. Loss of collectrin, an angiotensin-converting enzyme 2 homolog, uncouples endothelial nitric oxide synthase and causes hypertension and vascular dysfunction

    DEFF Research Database (Denmark)

    Cechova, Sylvia; Zeng, Qing; Billaud, Marie

    2013-01-01

    Collectrin is an orphan member of the renin-angiotensin system and is a homolog of angiotensin-converting enzyme 2, sharing ≈50% sequence identity. Unlike angiotensin-converting enzyme 2, collectrin lacks any catalytic domain. Collectrin has been shown to function as a chaperone of amino acid tra...

  19. Cytoplasmic male sterility in Petunia hybrida : a structural and histochemical analysis

    NARCIS (Netherlands)

    Bino, R.J.

    1986-01-01

    This thesis presents an analysis of the structural and histochemical aspects of cytoplasmic male sterility (cms) in Petuniahybrida . In petunia and in other crops, cms is the most commonly used tool for hybrid seed production. Application of the trait

  20. [Morphological and histochemical analysis of mucous membrane transformation of the artificial urinary bladder].

    Science.gov (United States)

    Iatsyna, A I; Vitruk, Iu V; Vernigorodskiĭ, S V; Stakhovskiĭ, A É; Emets, A I; Blium, Ia B; Stakhovskiĭ, É A

    2011-01-01

    The performed morphologic study of transformation mechanisms of the intestinal reservoir mucous membrane using modern immunochemical and histochemical methods revealed three morphologic phases of enteric epithelium adaptation; reactive-inflammatory, compensatory-protective and atrophic. Enteric urinary reservoir preserves the cambial apparatus typical for the enteric phenotype with an inherent apoptosis activity and proliferation which confirms a low level of tumor transformation risk.

  1. Histochemical and morphometric characteristics of muscle fibres: breeds and muscles comparison

    Directory of Open Access Journals (Sweden)

    G. Toscano Pagano

    2011-03-01

    Full Text Available Fibres histochemical characteristics seem affect the meat quality obtained from the skeletal muscles. In fact, the red fibres metabolise and store more lipid than the white ones (Ashmore et al., 1972, so important meat characteristics could be influenced by the muscle fibre type, as recently investigated (Morita et al., 2000; Ozawa et al., 2000; Vestergaard et al., 2000a, 2000b....

  2. Cytoplasmic male sterility in Petunia hybrida. A structural and histochemical analysis.

    NARCIS (Netherlands)

    Bino, R.J.

    1986-01-01

    This thesis presents an analysis of the structural and histochemical aspects of cytoplasmic male sterility (cms) in Petuniahybrida . In petunia and in other crops, cms is the most commonly used tool for hybrid seed production. Application of the trait makes hybrid seed production possi

  3. Classification of the intrafusal muscle fibres in the frog muscle spindle: histochemical and immunofluorescent studies.

    OpenAIRE

    Yoshimura, A; Fujitsuka, N; Sokabe, M; Naruse, K; Nomura, K; Diwan, F H; Ito, F

    1990-01-01

    Intrafusal muscle fibres from bull-frog semitendinosus, iliofibularis and sartorius muscles were classified into three types using the histochemical, immunofluorescent and morphological characteristics, with reference to the extrafusal muscle fibres, which were classified into five types in accordance with Rowlerson & Spurway (1988). Immunofluorescent reactions with antibodies against slow or fast myosins obtained from anterior or posterior latissimus dorsi muscles (ALD or PLD), respectively,...

  4. [Morphological and histochemical effects of subthreshold laser therapy on the chorioretinal complex].

    Science.gov (United States)

    Fedoruk, N A; Fedorov, A A; Bol'shunov, A V

    2013-01-01

    A variety of subthreshold laser methods that enable selective and precise treatment of chorioretinal microstructures, thus decreasing the risk of negative changes in retina and choriocapillaries, is used in current ophthalmological practice. This comparative pathomorphological and histochemical study allowed characterizing structural changes and functional activity of chorioretinal cells after application of different subthreshold laser technologies.

  5. Cytoplasmic male sterility in Petunia hybrida : a structural and histochemical analysis

    NARCIS (Netherlands)

    Bino, R.J.

    1986-01-01

    This thesis presents an analysis of the structural and histochemical aspects of cytoplasmic male sterility (cms) in Petuniahybrida . In petunia and in other crops, cms is the most commonly used tool for hybrid seed production. Application of the trait m

  6. Histochemical characterization of cytotoxic brain edema. Potassium concentrations after cerebral ischemia and during the postmortem interval.

    Science.gov (United States)

    Oehmichen, M; Ochs, U; Meissner, C

    2000-08-01

    Cytotoxic edema is a phenomenon of the ischemically damaged brain. In the present study we tested a histochemical method that detects this phenomenon based on potassium (K+) levels in the brain. In a first series focal cerebral ischemia was induced by arterial occlusion in 23 gerbils (Meriones unguiculatus). After survival times of 30, 60 and 120 min, the animals were killed and brain section histochemically stained for potassium and quantitatively evaluated with a morphometric method. The results were compared with those using physicochemical techniques. A distinct K+ depletion could be demonstrated in the area of the focal ischemia within a survival time of 30 min, the depletion growing thereafter with increasing survival time. In a second series histochemical and chemical methods were used to study the stability of K+ levels in undamaged brains of 15 healthy rats during postmortem intervals of 2.5 and 5 h. Within these intervals K+ levels were clearly depleted, apparently as a result of cerebral spinal fluid (CSF) diffusion. Even if neuronal injury can be demonstrated histochemically after very brief survival times of about 30 min, postmortem storage of the cadavers rendered detection impossible due to electrolyte and water diffusion. In autoptic human cases, therefore, this technique is of no practical utility in detecting cytotoxic brain edema in postmortem tissue.

  7. Histochemical and ultrastructural studies on the calcareous corpuscles and eggs of Taenia taeniaeformis and Dipylidium caninum.

    Science.gov (United States)

    Khalifa, Refaat M A; Mazen, Nawal A M; Marawan, Aziza M A; Thabit, Hasnaa T M

    2011-08-01

    Calcareous corpuscles were noticed by several previous workers to be present in larval and adult cestodes without knowing their function. However, nothing was mentioned in the available literature about distribution of these corpuscles and their density, structure and composition in different parts of the body of different cestodes. Hence, in the present work, a comparative study of their distribution, density, histochemical and ultrastructural characters in different parts of the body was performed in Taenia taeniaeformis and Dipylidium caninum. Due to the presence of the eggs in their gravid segments, their histochemical and ultrastructural characteristics were also studied. It was found that the size, location and density of the calcareous bodies were different in different body parts of the same and the other cestode. Histochemically, the main component of these corpuscles was calcium; while other constituents as polysaccharides, lipids, protrins and mucopolysaccharides were found in their outer rim. Ultrastructurally, they were quite similar in the two studied cestodes and different stages of their development were exhibited. Histochemically, the eggs of both cestodes were similar in their contents. However, some ultrastructural differences have been demonstrated particularly in relation to the size and shape of the rods in the embryophore and the structures in between the embryophore and onchosphere.

  8. Histochemical and enzymatic differences in skeletal muscle from Svalbard reindeer during the summer and winter

    Directory of Open Access Journals (Sweden)

    K.-H. Kiessling

    1986-06-01

    Full Text Available Enzyme activities and fibre properties in four muscles from Svalbard reindeer, collected during the summer, have been compared with corresponding muscles during the winter. In two muscles, gluteobiceps and semimembranosus, oxidative capacity is higher in winter than in summer; in the other two muscles, semitendinosus and longissimus dorsi, there is no difference with time of the year. The capacity to oxidize fatty acids is low in winter compared with summer, especially in semitendinosus and longissimus. These changes are similar in both sexes. Histochemical studies of the three main fibre types, I (BetaR, HA (°cR and IIB (aW, from the four muscles show that in male reindeers the muscle fibres are narrower at the end of the winter season than during the summer. The decrease of muscle tissue amounts to about one third of the total volume (33%, of which I accounts for 5%, IIA for 2% and IIB for 26%. The results indicate that the Svalbard reindeer use lean tissue in general, and IIB fibres in particular, in order to survive the hostile arctic winter period at Svalbard.Histokemiska och enzymatiska skillnader i skelettmuskel från Svalbardren mellan sommar och vinter.Abstract in Swedish / Sammandrag: Enzymaktiviteter och fiberegenskaper i fyra av Svalbardrenens muskler, insamlade under sommaren, har jåmforts med motsvarande muskler insamlade under vintern. I två muskler, gluteobiceps och semimembranosus, år oxidativa kapaciteten hogre under vintern an under sommaren; i de andra två musklerna, semitendinosus och longissimus dorsi, foreligger ingen skillnad i detta avseende. Kapaciteten att oxidera fettsyror år låg under vintern jåmfort med sommaren, speciellt i semitendinosus och longissimus. Inga konsskillnader foreligger i dessa avseenden. Histokemiska studier av de tre huvudtyperna av muskelfibrer, fiR (I, ocR (IIA och (IIB, från de fyra musklerna visar att hos handjuren år fibrerna tunnare vid slutet av vintersåsongen jåmfort med

  9. Effects of gaseous hydrogen fluoride on oxidative enzymes of Pelargonium zonale leaves

    Energy Technology Data Exchange (ETDEWEB)

    Poovaiah, B.W.; Wiebe, H.H.

    1971-10-01

    Changes in peroxidase and cytochrome oxidase enzymes were established histochemically in hydrogen fluoride fumigated leaves of Pelargonium zonale. Highest peroxidase and cytochrome oxidase activities were localized near the injured areas of fumigated leaves, and the greatest increase was observed in the phloem region.

  10. Carbohydrate specificity of lectin, purified from the fruiting bodies of Mycena pura /Fr./ Kumm. and its use in histochemical investigation

    Directory of Open Access Journals (Sweden)

    Ambarova N. O.

    2009-12-01

    Full Text Available Aim. The purpose of this investigation was to research carbohydrate specificity of a new lectin from fruiting body of Mycena pura and possibilities of its application in histochemical studies. Methods. The lectin has been purified by affinity chromatography on «îvomucine». The lectin carbohydrate specificity has been determined by a reaction of inhibiting haemagglutination by haptens. Histological materials were fixed in 4 % neutral formalin solution. Alkaline phosphatase was revealed in the cryostat unfixed microscopical sections. Results. The lectin yield from fresh fruit bodies of raw material was 9 mg/kg. Mol. mass of the lectin is 40 kDa. The lectin poorly interacted with D-glucose and D-mannose in contrast to lectins from Pisum sativum and Leucojum vernum. The peculiarity of this lectin is its strong interaction with alkaline phosphatase, the highest among twenty tested lectins. However, the receptors for Mycena lectin binding in mammalian tissues are not limited by this enzyme being presented also by glycoconjugates of another structure, as it was shown for fetus calf small intestine and kidney of rat. Conclusions. An important role in the lectin interaction with glycoproteins probably belongs to the disaccharide links of GlcNAcb(1-2Mana(1-6 or GlcNAcb(1- 2Mana(1-2, which not necessarily are terminal

  11. Hereditary fructose intolerance mimicking a biochemical phenotype of mucolipidosis: A review of the literature of secondary causes of lysosomal enzyme activity elevation in serum.

    Science.gov (United States)

    Ferreira, Carlos R; Devaney, Joseph M; Hofherr, Sean E; Pollard, Laura M; Cusmano-Ozog, Kristina

    2017-02-01

    We describe a patient with failure to thrive, hepatomegaly, liver dysfunction, and elevation of multiple plasma lysosomal enzyme activities mimicking mucolipidosis II or III, in whom a diagnosis of hereditary fructose intolerance (HFI) was ultimately obtained. She presented before introduction of solid foods, given her consumption of a fructose-containing infant formula. We present the most extensive panel of lysosomal enzyme activities reported to date in a patient with HFI, and propose that multiple enzyme elevations in plasma, especially when in conjunction with a normal plasma α-mannosidase activity, should elicit a differential diagnosis of HFI. We also performed a review of the literature on the different etiologies of elevated lysosomal enzyme activities in serum or plasma. © 2016 Wiley Periodicals, Inc.

  12. Histochemical and immunohistochemical studies of the gonads and paramesonephric ducts of male and female hatchlings of loggerhead sea turtles (Caretta caretta).

    Science.gov (United States)

    Sarı, F; Kaska, Y

    2016-08-01

    The sex of neonatal sea turtles is difficult to determine, because neonates lack heteromorphic sex chromosomes and dimorphic external characteristics; internal dimorphic morphology is defined at hatching. We used histochemical staining and made measurements in the gonads and paramesonephric ducts (PD) of both sexes to determine structural differences in female and male loggerhead sea turtle (Caretta caretta) hatchlings. We detected differences in the gonads and PD between the sexes including the amounts of mucopolysaccharides, collagen and elastic fibers. We determined that the thickness of the gonadal cortex and the diameter of the PD lumen are reliable sex-specific characteristics. We also assessed immunolocalization of aromatase, an enzyme complex that converts androgens to estrogens, and found differences in the localization and intensity of aromatase immunostaining in the gonads and PD of female and male hatchlings. Comprehensive studies of the sexual differences of sea turtles are important for conservation programs.

  13. Histological and histochemical evaluations on the effects of high incubation temperature on the embryonic development of tibial growth plate in broiler chickens.

    Science.gov (United States)

    Oznurlu, Yasemin; Sur, Emrah; Ozaydin, Tugba; Celik, Ilhami; Uluisik, Deniz

    2016-02-01

    The effects of experimentally induced high incubation temperature on the embryonic development of growth plate of the chicken were investigated by means of histological and enzyme histochemical methods. In the experiments, 250 fertile eggs of Ross-308 broiler strain were divided into two groups, the control eggs were maintained under optimal conditions (37.8°C and 65% ± 2% relative humidity, rh) during the whole incubation period. Heat-stress imposed eggs were maintained under normal conditions (37.8°C and 65% ± 2% rh) until the 10th day of incubation, and then, continuously (24 h per day) exposed to high temperature (38.8°C and 65% ± 2% rh). Tissue samples were taken from 10 animals of each group at the 11th, 13th, 15th, 18th, 21st days of incubation. Tissue samples were processed by enzyme histochemical methods in addition to routine histological techniques. The relative tibia weights and tibia length were lower in the heat-stress group compared to the control group. The results of the measurements of the growth plate showed that the proliferative zone was narrowed whereas, the transitional and hypertrophic zone were thickened in the heat stress group. Alkaline phosphatase (ALP) density was significantly decreased in the heat-stress group compared to the control group. In conclusion, bone formation and growth plate formation are crucial for embryo development and 1°C higher from optimum may increase the incidence of skeletal disorders and leg problems in broiler chickens which is one of the major animal welfare concerns for the poultry industry. © 2016 Wiley Periodicals, Inc.

  14. Bile acid malabsorption or disturbed intestinal permeability in patients treated with enzyme substitution for exocrine pancreatic insufficiency is not caused by bacterial overgrowth

    DEFF Research Database (Denmark)

    Madsen, Jan Lysgård; Graff, Jesper; Philipsen, Else Kirstine;

    2003-01-01

    enzyme replacement therapy, were studied. The prevalence of bacterial overgrowth was evaluated by means of a hydrogen and methane breath test with glucose. Gamma camera scintigraphy after intake of 75Se-homocholic acid taurine (75Se-HCAT) was used to evaluate bile acid absorption capacity. Intestinal......INTRODUCTION: In some patients with severe exocrine pancreatic insufficiency, enzyme replacement therapy will not lead to clinical improvement or reduction of steatorrhea. Therefore, other mechanisms separately or in interplay with reduced enzyme secretion might be responsible for malabsorption...... in these patients. AIMS: To evaluate the prevalence of bacterial overgrowth, bile acid absorption capacity, and intestinal permeability in a group of patients with well-characterized exocrine pancreatic insufficiency. METHODOLOGY: Eleven men with severe exocrine pancreatic insufficiency, of whom 10 were receiving...

  15. Pectic enzymes

    NARCIS (Netherlands)

    Benen, J.A.E.; Voragen, A.G.J.; Visser, J.

    2003-01-01

    The pectic enzymes comprise a diverse group of enzymes. They consist of main-chain depolymerases and esterases active on methyl- and acetylesters of galacturonosyl uronic acid residues. The depolymerizing enzymes comprise hydrolases as wel as lyases

  16. Pectic enzymes

    NARCIS (Netherlands)

    Benen, J.A.E.; Voragen, A.G.J.; Visser, J.

    2003-01-01

    The pectic enzymes comprise a diverse group of enzymes. They consist of main-chain depolymerases and esterases active on methyl- and acetylesters of galacturonosyl uronic acid residues. The depolymerizing enzymes comprise hydrolases as wel as lyases

  17. Enzyme assays.

    Science.gov (United States)

    Reymond, Jean-Louis; Fluxà, Viviana S; Maillard, Noélie

    2009-01-07

    Enzyme assays are analytical tools to visualize enzyme activities. In recent years a large variety of enzyme assays have been developed to assist the discovery and optimization of industrial enzymes, in particular for "white biotechnology" where selective enzymes are used with great success for economically viable, mild and environmentally benign production processes. The present article highlights the aspects of fluorogenic and chromogenic substrates, sensors, and enzyme fingerprinting, which are our particular areas of interest.

  18. Taxonomic value of histochemical features of the style in early lineages of Asteraceae

    Directory of Open Access Journals (Sweden)

    Marcelo Paulo Hernández

    2015-12-01

    Full Text Available In this paper we examine lipophilic substances, tannins, proteins, reducing carbohydrates, starch, and oxalate salts in early lineages of Asteraceae to test if they are a useful complement to classification studies. Styles of mature flowers of 44 species were processed according to plant histochemical techniques and observed by means of a light microscope. Lipophilic compounds were always present in the cuticle and cell walls, and in some species were located inside the cells. Proteins were located in the style cell walls and cell interior of most of the analyzed species. No tannins were found. In general, there is a correlation in the presence of reducing sugars and proteins. Starch granules and oxalate salts were found in the style parenchyma of a few species. Cytoplasmic content of lipophilic compounds, proteins, and reducing sugars predominates in Barnadesioideae and Nassauvieae (Mutisioideae, and histochemical similarities were found among members of Gochnatioideae

  19. Mercury-selenium interactions in relation to histochemical staining of mercury in the rat liver

    DEFF Research Database (Denmark)

    Baatrup, E; Thorlacius-Ussing, O; Nielsen, H L

    1989-01-01

    of the radioactively labelled Hg compounds showed that the chemical form of mercury, either organic or inorganic, was preserved from its administration to its deposition in the liver. Light and electron microscopy demonstrated that no silver enhancement of Hg occurred when MeHg alone was present in the sections......Selenium has been suggested to enhance the histochemical staining of mercury when sections of tissue are subjected to the silver-enhancement method. In the present study, histochemical staining patterns of mercury in tissue sections of rat livers were compared with the actual content of organic...... and inorganic Hg in the livers, in both the presence and the absence of Se. Rats were injected intravenously with 5 micrograms of Hg g-1 body weight as methyl [203Hg] mercury chloride (MeHg) or as [203Hg]mercuric chloride (Hg2+). After 2 h, half the rats received an additional intraperitoneal injection of 2...

  20. Case report. Sclerosing peritoneal mesothelioma in a dog: histopathological, histochemical and immunohistochemical investigations

    Directory of Open Access Journals (Sweden)

    Anna Rita D'Angelo

    2014-12-01

    Full Text Available Mesotheliomas are rare neoplasm affecting on rare occasions both animals and humans and which arise from the mesothelial cells lining the coelomic cavities. We report herein the histopathological, histochemical and immunohistochemical findings in a dog affected by sclerosing peritoneal mesothelioma, a rare variant of canine mesothelioma, and submitted to laparotomy in December 2012 (Teramo, Italy. Our data confirm that mesothelioma still represents a diagnostic challenge and that immunohistochemistry can be extremely useful as supportive diagnostic technique.

  1. Organogenesis of the digestive tract in the white seabream, Diplodus sargus. Histological and histochemical approaches

    OpenAIRE

    2003-01-01

    The ontogeny of the digestive tract of the white seabream, Diplodus sargus during the larval development up to day 45 post-hatching (dph) has been studied using histological and histochemical techniques. The oesophageal goblet cells appeared around 6 dph and contained neutral and acid mucosubstances (PAS/diastase-PAS and Alcian Blue pH 2.5 positive reactions). An incipient stomach can be distinguished from 2 dph but the first sign of gastric gland developme...

  2. Histochemical and morphological characteristics of the levator ani muscle in rats

    OpenAIRE

    2011-01-01

    The current study aimed to analyze the histochemical and morphological characteristics of the levator ani muscle in rats. For this, we used 10 Wistar rats (5 males and 5 females), weighing between 200 and 765g. The animals were dissected fresh and in formalin for the levator ani muscle anatomical observation. Muscle fragments were collected and frozen in n-Hexane previously cooled in liquid nitrogen. Then, the muscles were transferred to a microtome cryostat (HM 505 E Microm), being fixed in ...

  3. An anatomical and histochemical examination of the pituitary gland of carp (Cyprinus carpio)

    OpenAIRE

    EKİCİ, Aygül; TİMUR, Metin

    2014-01-01

    The present study was carried out for the purpose of studying the anatomical and the histochemical structure of the pituitary gland of the common carp, Cyprinus carpio. Anatomically, the shape of the gland has been observed to be round-oval, looking like an acorn. The pituitary gland consists of the adenohypophysis and neurohypophysis parts. Microscopically, the adenohypophysis consists of anterior (pars distalis) and posterior (pars intermedia) parts. The second lobe of the gland, called the...

  4. [Selective histochemical identification of neuronal cell populations using fucose-specific lectins].

    Science.gov (United States)

    Akkuratov, E G; Nozdrachev, A D

    2004-01-01

    We studied lectin histochemical properties of structures of caudal ganglia of the vagus nerve and ganglion of the trigeminal nerve in white rats using fucose-specific conjugates to peroxidase. Morphological samples were processed on a computer video analyzer. Metrical and optical indices of the afferent neurons were analyzed. The obtained data demonstrate different topography of glycoconjugates in the afferent ganglia. Application of recent image processing techniques allows revealing neuron populations in afferent ganglia of rats undetectable by standard morphological techniques.

  5. Locus ceruleus neurons in people with autism contain no histochemically-detectable mercury

    OpenAIRE

    2015-01-01

    Exposure to environmental mercury has been proposed to play a part in autism. Mercury is selectively taken up by the human locus ceruleus, a region of the brain that has been implicated in autism. We therefore looked for the presence of mercury in the locus ceruleus of people who had autism, using the histochemical technique of autometallography which can detect nanogram amounts of mercury in tissues. In addition, we sought evidence of damage to locus ceruleus neurons in autism by immunostain...

  6. Inactivation of the HR6B ubiquitin-conjugating DNA repair enzyme in mice causes male sterility associated with chromatin modification.

    NARCIS (Netherlands)

    J. van Klaveren; J. de Wit (Jan); C.G. van Gurp; M.H.M. Koken (Marcel); M. Vermey; J.H. van Roijen (Jan Herman); J.T.M. Vreeburg (Jan); W.M. Baarends (Willy); D. Bootsma (Dirk); J.A. Grootegoed (Anton); J.H.J. Hoeijmakers (Jan); H.P. Roest (Henk)

    1996-01-01

    textabstractThe ubiquitin-conjugating yeast enzyme RAD6 and its human homologs hHR6A and hHR6B are implicated in postreplication repair and damage-induced mutagenesis. The yeast protein is also required for sporulation and may modulate chromatin structure via histone ubiquitination. We report the ph

  7. Anatomical and histochemical analysis of Dysphania ambrosioides supported by light and electron microscopy

    Directory of Open Access Journals (Sweden)

    Rafaela D. Sá

    Full Text Available ABSTRACT Dysphania ambrosioides (L. Mosyakin & Clemants (syn: Chenopodium ambrosoides L., Amaranthaceae, popularly known as “mastruz”, is an herb widely used in Brazil as anthelmintic. To contribute to the knowledge about medicinal plants, a microscopic analysis was accomplished to describe the main anatomical characters of root, stem, petiole and leaf blade of D. ambrosioides and histochemical tests were performed on the leaf blade. Cross-sections were obtained, by hand, for microscopic analysis of root, stem, petiole and leaf blade; to the leaf blade were still made paradermal sections, scanning electron microscopy analysis, maceration and histochemical tests. The main characters useful in the identification of the plant were: anomalous secondary thickening in the root and stem; presence of idioblasts containing crystal sand in the root, stem, petiole and leaf blade; in these there are also idioblasts with druses; presence of non-glandular and glandular trichomes in the stem, petiole and leaf blade; stomata on the stem, petiole and leaf blade, identified in these as anomocytic and anisocytic; dorsiventral mesophyll and collateral vascular bundles. Maceration revealed that the vessel elements are helical type. Through the histochemical tests, it was evidenced the presence of lipophilic substances, essential oils, oleoresins, phenolic compounds, starch, lignin and calcium oxalate crystals. This work provides support to the quality control of the species.

  8. Nitrergic neurons during early postnatal development of the prefrontal cortex in the rat: histochemical study.

    Science.gov (United States)

    Hvizdosova, Natalia; Tomasova, Lenka; Bolekova, Adriana; Kolesar, Dalibor; Kluchova, Darina

    2014-06-01

    The presence of nitrergic cells in the prefrontal cortex has been confirmed, however little is known about the postnatal development of these cells. Nitrergic neurons were studied histochemically by using NADPH-diaphorase staining in the prefrontal cortex of male Wistar rats from postnatal day 7-21 (P7-21). Neuronal NADPH-diaphorase is a nitric oxide synthase that provides a specific histochemical marker for neurons producing nitric oxide (NO). NO acts as a neurotransmitter and intracellular signaling molecule in the nervous system. We observed in 7 day old rats NADPH-d containing neurons that were intensely stained. These neurons were bipolar with a short dendrite with average length of 23 μm. During the second postnatal week, the neurons were mainly bipolar and were rarely multipolar. By P14 the cells were located primarily in cortical layers III-VI. Nitrergic neurons of the 21 day old rats were histochemically identified as multipolar cells with long radial extending dendrites. Dendrites of neurons in 14 and 21 day old rats were a similar length with an average of 57 μm. These results suggest that nitrergic neurons differentiate during a relatively short period of time and reach their structural maturity by the end of the second week of postnatal development.

  9. Analysis of gene expression changes, caused by exposure to nitrite, in metabolic and antioxidant enzymes in the red claw crayfish, Cherax quadricarinatus.

    Science.gov (United States)

    Jiang, Qichen; Zhang, Wenyi; Tan, Hongyue; Pan, Dongmei; Yang, Yuanhao; Ren, Qian; Yang, Jiaxin

    2014-06-01

    We evaluated the effect of acute exposure to nitrite on expression of antioxidant and metabolic enzyme genes in gill tissue of advanced juvenile Cherax quadricarinatus. A 48h nitrite exposure was conducted, using four test concentrations (NO2-N=0.5, 1, 1.5 and 2mg L(-1)) plus a control group. The relative mRNA expression of mitochondrial manganese superoxide dismutase (mMnSOD), cytosolic MnSOD (cMnSOD), extracellular copper/zinc SOD (exCu/ZnSOD), catalase (CAT), glutathione S-transferase (GST), arginine kinase (AK), glutamate dehydrogenase (GDH), mitochondrial malate dehydrogenase (mMDH), Na(+)/K(+)-ATPase α-subunit and phosphoenolpyruvate carboxykinase (PEPCK) in gill tissue was measured. Significantly increased mRNA expression was observed for all the antioxidant enzymes after 12 and 24h. After 48h, they all decreased at high nitrite concentrations. The gene expression levels of AK, GDH, mMDH and Na(+)/K(+)-ATPase α-subunit showed similar trends as the antioxidant enzymes. Significant depression of gene expression levels of PEPCK occurred throughout the experimental time at high nitrite concentrations. The results indicated that nitrite could induce oxidative and metabolic stress in C. quadricarinatus, in a time dependent manner, which suggests they could be helpful in predicting sublethal nitrite toxicity and useful in environmental monitoring studies.

  10. Cytological And Histochemical Studies On Rat Liver And Pancreas During Progression Of Streptozotocin Induced Diabetes And Possible Protection Using Certain Natural Antioxidants

    Directory of Open Access Journals (Sweden)

    Hanaa F. Waer*, Seham A. Helmy

    2012-07-01

    Full Text Available Background: Diabetes mellitus is a major endocrine disorder and growing health problem in most countries. Diabetes manifested by experimental animal models exhibits high oxidative stress due to persistent and chronic hyperglycemia which increases the generation of free radicals, streptozotocin (STZ provides an animal model of type 1 diabetes. Thereby depleting the activities of antioxidative defense systems with alteration of antioxidant activities of enzymes such as green tea and curcumin . Aim : Biochemical histological and histochemical investigations were carried on to revel the effect of STZ on the liver and pancreas cells. Natural antioxidants were used as a new way for ameliorating diabetic effect on the cellsMaterial and methods: Diabetes was induced by a single intraperitoneal injection of freshly prepared STZ dissolved in 0.05M of sodium citrate buffer, pH = 4.6, (STZ; 45 mg/kg B.wt..Three days after degeneration of beta cells, diabetes was induced in all animals. After induction of diabetes, diabetic and normal animals were kept in metabolic cages separately. Green tea (EGCG and curcumin are used as natural antioxidants to improve the disorders and structural changes induced by STZ. Cellular and histochemical investigations were carried on the changes induced in the pancreatic and hepatic tissues. Body weight, level of serum glucose and insulin were calculated in the control and treated groups. For detecting the degeneration of both hepatocytes and pancreatic cells of diabetic rats, tissue samples from diabetic and treated rats were collected and pathologically examined. Results: The present investigations reveled that there was a detectable amelioration on the injures induced by STZ on both hepatocytes and pancreatic cells using green tea or curcumin with a detectable dose level. Also it can be observed that the ameliorated effect induced was a time dependant. Conformation of these results from histochemical detection of

  11. Cleft lip: a histochemical and ultrastructural analysis of lip muscles.

    Science.gov (United States)

    Raposio, E; Cella, A; Panarese, P; Renzi, M; Caregnato, P; Barabino, P; Faggioni, M; Gualdi, A; Santi, P L

    1999-10-01

    In order to evaluate the pathogenesis of cleft-lip in relation to both the anatomical and structural anomalies of the mesenchymal tissues, the authors concluded that the presence of structural anomalies in the examined tissues could not explain the malformation, but might be a consequence of it. Delayed muscular development, asymmetrical distribution of the muscular fibres and their anomalous insertion suggest that the anatomical/functional loss clinically detectable in the orbicular muscle could be the result of a perinatal dysmorphological process rather than of a simple mesenchymal hypoplasia. Schendel et al. suggested that a metabolic defect in the mitochondrial function could cause a deficiency in cell migration and proliferation responsible for the malformation in question. To establish whether the pathogenesis of the cleft-lip is associated with an alteration in mitochondrial functionality, eight patients affected by unilateral cleft-lip were subjected to a biopsy of the orbicular muscle during the course of reparative surgery. The results obtained showed: 1) a great variation in the size of muscle fibres; 2) the absence of ragged red fibres; 3) a normal oxidative function in the muscle fibres examined; 4) the absence of typologically significant groupings positive for myofibral ATPases. Furthermore, the morphology of the mitochondria was preserved in all cases and neither inclusions nor morphological or volumetric changes were detected. This preliminary data did not confirm the constant presence of mitochondrial pathology responsible for the malformation in question. In our opinion, the growth deficiency of the maxillary segment could be ascribed to the cicatrization of the surgical repair of the cleft-lip.

  12. Histochemical evidence of osteoclastic degradation of extracellular matrix in osteolytic metastasis originating from human lung small carcinoma (SBC-5) cells.

    Science.gov (United States)

    Li, Minqi; Amizuka, Norio; Takeuchi, Kiichi; Freitas, Paulo H L; Kawano, Yoshiro; Hoshino, Masaaki; Oda, Kimimitsu; Nozawa-Inoue, Kayoko; Maeda, Takeyasu

    2006-02-01

    The aim of this study was to assess the dynamics of osteoclast migration and the degradation of unmineralized extracellular matrix in an osteolytic metastasis by examining a well-standardized lung cancer metastasis model of nude mice. SBC-5 human lung small carcinoma cells were injected into the left cardiac ventricle of 6-week-old BALB/c nu/nu mice under anesthesia. At 25-30 days after injection, the animals were sacrificed and their femora and/or tibiae were removed for histochemical analyses. Metastatic lesions were shown to occupy a considerable area extending from the metaphyses to the bone marrow region. Tartrate resistant acid phosphatase (TRAPase)-positive osteoclasts were found in association with an alkaline phosphatase (ALPase)-positive osteoblastic layer lining the bone surface, but could also be localized in the ALPase-negative stromal tissues that border the tumor nodules. These stromal tissues were markedly positive for osteopontin, and contained a significant number of TRAPase-positive osteoclasts expressing immunoreactivity for CD44. We thus speculated that, mediating its affinity for CD44, osteopontin may serve to facilitate osteoclastic migration after their formation associated with ALPase-positive osteoblasts. We next examined the localization of cathepsin K and matrix metallo-proteinase-9 (MMP-9) in osteoclasts. Osteoclasts adjacent to the bone surfaces were positive for both proteins, whereas those in the stromal tissues in the tumor nests showed only MMP-9 immunoreactivity. Immunoelectron microscopy disclosed the presence of MMP-9 in the Golgi apparatus and in vesicular structures at the baso-lateral cytoplasmic region of the osteoclasts found in the stromal tissue. MMP-9-positive vesicular structures also contained fragmented extracellular materials. Thus, osteoclasts appear to either select an optimized function, namely secreting proteolytic enzymes from ruffled borders during bone resorption, or recognize the surrounding extracellular

  13. Observations on pathological and histochemical changes in piglet livers infected with Taenia saginata asiatica

    Institute of Scientific and Technical Information of China (English)

    LINGHU Yan; ZHU Wujun; BAO Huai'en; CHEN Yan

    2007-01-01

    To study the pathological and histochemical characteristies of lesions in piglet livers infected with Taenia saginata asiatica (T.saginata asiatica)throughout the different stages,piglets were fed with eggs of T.saginata asiatica and raised in isolation in an animal center to establish the T.saginata asiatica infection model with normal piglets as control.The pathological changes in the piglet livers were observed after the infection using liver sections stained with hematoxylin and eosin.Histochemical methods were used to check the changes in lipid,glycogen and protein content in the liver.The data collected by image analysis were analyzed statistically with Statistical Package for the Social Science.The results show that T.saginata asiatica-exposed piglets were indeed infected.Inflammatory reactions began on the fourth day and progressed rapidly.Kupffer cell hyperplasia,hepatic hydropic degeneration and ballooning degeneration were found in the 10th-20th days after infection.Hepatic central veins and hepatic sinusoids were dilated and congested.Spotty necrosis occurred in some local liver tissues.In the 40th-60th days,granulomatous reactions and mild hepatocirrhosis were the main lesions.In the 70th-80th days,hepatocirrhosis and bile duct proliferation were observed in the liver.In the difierent stages,lipid drops were increased while glycogen and protein levels were decreased to some degree.There was a significant difierence in metabolism between the infected group and the control group(P<0.01).It is concluded that pigs are the favorable intermediate host of T.saginata asiatica and its infection can result in serious pathological and histochemical lesions in host organs.

  14. Mercury-selenium interactions in relation to histochemical staining of mercury in the rat liver.

    Science.gov (United States)

    Baatrup, E; Thorlacius-Ussing, O; Nielsen, H L; Wilsky, K

    1989-02-01

    Selenium has been suggested to enhance the histochemical staining of mercury when sections of tissue are subjected to the silver-enhancement method. In the present study, histochemical staining patterns of mercury in tissue sections of rat livers were compared with the actual content of organic and inorganic Hg in the livers, in both the presence and the absence of Se. Rats were injected intravenously with 5 micrograms of Hg g-1 body weight as methyl [203Hg] mercury chloride (MeHg) or as [203Hg]mercuric chloride (Hg2+). After 2 h, half the rats received an additional intraperitoneal injection of 2 micrograms of Se g-1 body weight as sodium [75Se]selenite. All the rats were killed 1 h later. Homogenized liver samples were prepared for mercury analysis by two different methods: alkaline digestion and ultrasonic disintegration. Quantitative chemical analysis based on benzene extraction of the radioactively labelled Hg compounds showed that the chemical form of mercury, either organic or inorganic, was preserved from its administration to its deposition in the liver. Light and electron microscopy demonstrated that no silver enhancement of Hg occurred when MeHg alone was present in the sections of tissue, whereas MeHg accompanied by Se induced a moderate deposition of silver grains. In contrast, sections containing Hg2+ alone yielded some staining, and the addition of Se increased the staining dramatically. The results of the present study show that acute selenite pretreatment is a prerequisite for the histochemical demonstration of methyl mercury, and greatly increases the staining of inorganic mercury when applying the silver-enhancement method.

  15. Electron miscroscopy and histochemical studies on four Egyptian helminthes eggs of medical importance.

    Science.gov (United States)

    Mahmoud, Lily H; el-Alfy, Nadia M

    2003-04-01

    By SEM the Fasciola gigantica egg is ovoid with a small knob like operculum, while the egg of Heterophyes heterophytes is broad oval with the operculum more tapering. The egg shell of fertilized Ascaris lumbricoides has interconnected ridges and peak-like projections, while the egg of Enterobius vermicularis is flattened with a thicker margin at the curved side. By TEM, Fasciola egg shell consists of fine reticulum fibrils of three layers. The outer lipoprotein of perivitelline membrane beneath which 2 membranes separated by inclusions, middle of protein globules and inner lipoprotein layer with minute electron-dense granules of melanin or polymer origin, in some parts of the shell giving the egg its brown coloration. The Heterophyes egg shell is more or less similar to that of Fasciola but lacking the minute electron-dense granules. The egg shell of Ascaris has outer ulterine layer with three consecutive layers, basal lipoprotein layer and the inner lipid or ascaroside layer which is the most resistant layer. The Enterobius egg shell consists of five layers, external uterine, internal uterine, vitelline, chitinous and lipid layer. Histochemically, Fasciola egg shell consists of nine amino-acids, and that of Heterophyes consists of ten amino acids. In Ascaris, the lipid layer characteristically consists 25% protein and 75% lipid. The histochemical examination of Enterobius as a detailed example, showed different degrees of reactions with mercuric bromophenol blue, diazotization coupling, Sakaguchi reaction, Sudan black and Mallory's triple stain. Temperature showed marked effect on eggs survival. Eggs of Fasciola and Heterophyes withstand more low temperatures but those of Ascaris and Enterobius withstand more high ones. There are marked correlations between the egg shell constitution, histochemical compositions on one hand and water permeability and egg dryness on the other hand. The results were photographed and discussed.

  16. Viability effects and not meoitic drive cause dramatic departures from Mendelian inheritance for malic enzyme in hybrids of Tigriopus californicus populations.

    Science.gov (United States)

    Willett, C S; Berkowitz, J N

    2007-05-01

    The genetic basis of post-zygotic reproductive isolation is beginning to be untangled in closely related species, but less is known about the genetics of reproductive isolation between divergent populations. Here, two genes encoding malic enzyme (ME) are isolated from the copepod Tigriopus californicus and their influence upon lowered viability in F(2) hybrids of genetically divergent populations is determined. Each ME gene has diverged extensively between T. californicus populations and one gene shows evidence for a recent selective sweep. Segregation patterns of genotypes for both ME genes in adult F(2) hybrids reveal dramatic departures from Mendelian inheritance, deviations that are not seen in F(2) nauplii implying that selection is acting during development based upon the genotype at these ME genes. These results imply that selection against deleterious gene combinations and not aberrant segregation (i.e. meiotic drive) is likely to lead to dramatic departures from Mendelian inheritance observed in these crosses.

  17. Improved histochemical methods for the examination of plastic-embedded human marrow

    Energy Technology Data Exchange (ETDEWEB)

    Moosavi, H.; Lichtman, M. A.; Donnelly, J. A.; Churukian, C. J.

    1979-01-01

    Improved methods for processing, sectioning, and staining plastic (glycol methacrylate) embedded human marrow biopsies are described. Marrow biopsies processed by this technique were compared with biopsies processed by the conventional paraffin embedding method. The plastic-embedded marrows provide better morphology enhancing diagnostic accuracy, permit assessment of bone as well as marrow, and allow histochemical analysis of biopsy specimen. Special stains including naphtol AS-D chloroacetate esterase, periodic acid Schiff (PAS), reticulin, and iron have been modified so that they are suitable for undecalcified, two microns thick, plastic-embedded human marrow biopsies.

  18. [Histochemical study of the digestive organs of rats after a flight on "Kosmos-605"].

    Science.gov (United States)

    Shubich, M G; Goriacheva, L L; Dudetskiĭ, V I; Lutsenko, N M; Mogil'naia, G M

    1977-01-01

    The histochemical study of the stomach, small and large intestines and pancreas of rats flown aboard the biosatellite Cosmos-605 as well as of synchronous and vivarium controls demonstrated a significant decline in the mucine producing capacity of epithelial cells of the stomach of the flight rats on the R + 1 day. The study showed an increased content of sialo- and sulphosaccharides in goblet cells of cryptae of large intestine and a reduced content of free cation protein in the acinar cells of the pancreas of flight rats. The changes were transient and disappeared by the R + 26 day.

  19. Histochemical studies on genetical control of hormonal enzyme inducibility in the mouse. VI. Effects of short term castration

    DEFF Research Database (Denmark)

    Kjaer, K; Kirkeby, S; Blecher, S R

    1983-01-01

    The regional histology and esterase activity of the mouse epididymis after 24, 48, and 72 hr castration is reported. Differential sensitivity to androgen deprivation among the various epithelial cell types is described, allowing of positive identification of the cell types previously observed...... to survive long-term castration. The possibility of an androgen binding protein, as described in the rat and rabbit, is suggested on morphological grounds. The epididymal body appears to contain a class of highly androgen sensitive cells that degenerate rapidly following castration and a second class...

  20. The Catalytic Function of Enzymes.

    Science.gov (United States)

    Splittgerber, Allan G.

    1985-01-01

    Discusses: structure of the enzyme molecule; active site; reaction mechanism; transition state; factors affecting enzyme reaction rates, concentration of enzyme; concentration of substrate; product concentration; temperature effects and pH effects; factors causing a lowering of activation energy; proximity and orientation effects; substrate strain…

  1. Mutations in B3GALT6, which encodes a glycosaminoglycan linker region enzyme, cause a spectrum of skeletal and connective tissue disorders.

    Science.gov (United States)

    Nakajima, Masahiro; Mizumoto, Shuji; Miyake, Noriko; Kogawa, Ryo; Iida, Aritoshi; Ito, Hironori; Kitoh, Hiroshi; Hirayama, Aya; Mitsubuchi, Hiroshi; Miyazaki, Osamu; Kosaki, Rika; Horikawa, Reiko; Lai, Angeline; Mendoza-Londono, Roberto; Dupuis, Lucie; Chitayat, David; Howard, Andrew; Leal, Gabriela F; Cavalcanti, Denise; Tsurusaki, Yoshinori; Saitsu, Hirotomo; Watanabe, Shigehiko; Lausch, Ekkehart; Unger, Sheila; Bonafé, Luisa; Ohashi, Hirofumi; Superti-Furga, Andrea; Matsumoto, Naomichi; Sugahara, Kazuyuki; Nishimura, Gen; Ikegawa, Shiro

    2013-06-01

    Proteoglycans (PGs) are a major component of the extracellular matrix in many tissues and function as structural and regulatory molecules. PGs are composed of core proteins and glycosaminoglycan (GAG) side chains. The biosynthesis of GAGs starts with the linker region that consists of four sugar residues and is followed by repeating disaccharide units. By exome sequencing, we found that B3GALT6 encoding an enzyme involved in the biosynthesis of the GAG linker region is responsible for a severe skeletal dysplasia, spondyloepimetaphyseal dysplasia with joint laxity type 1 (SEMD-JL1). B3GALT6 loss-of-function mutations were found in individuals with SEMD-JL1 from seven families. In a subsequent candidate gene study based on the phenotypic similarity, we found that B3GALT6 is also responsible for a connective tissue disease, Ehlers-Danlos syndrome (progeroid form). Recessive loss-of-function mutations in B3GALT6 result in a spectrum of disorders affecting a broad range of skeletal and connective tissues characterized by lax skin, muscle hypotonia, joint dislocation, and spinal deformity. The pleiotropic phenotypes of the disorders indicate that B3GALT6 plays a critical role in a wide range of biological processes in various tissues, including skin, bone, cartilage, tendon, and ligament.

  2. Immunohistochemical/histochemical double staining method in the study of the columnar metaplasia of the oesophagus

    Directory of Open Access Journals (Sweden)

    D. Cabibi

    2014-03-01

    Full Text Available Intestinal metaplasia in Barrett’s oesophagus (BO represents an important risk factor for oesophageal adenocarcinoma. Instead, few and controversial data are reported about the progression risk of columnar-lined oesophagus without intestinal metaplasia (CLO, posing an issue about its clinical management. The aim was to evaluate if some immunophenotypic changes were present in CLO independently of the presence of the goblet cells. We studied a series of oesophageal biopsies from patients with endoscopic finding of columnar metaplasia, by performing some immunohistochemical stainings (CK7, p53, AuroraA combined with histochemistry (Alcian-blue and Alcian/PAS, with the aim of simultaneously assess the histochemical features in cells that shows an aberrant expression of such antigens. We evidenced a cytoplasmic expression of CK7 and a nuclear expression of Aurora A and p53,  both in goblet cells of BO and in non-goblet cells of CLO, some of which showing mild dysplasia. These findings suggest that some immunophenotypic changes are present in CLO and they can precede the appearance of the goblet cells or can be present independently of them, confirming the conception of BO as the condition characterized by any extention of columnar epithelium. This is the first study in which a combined immunohistochemical/histochemical method has been applied to Barrett pathology.

  3. Protective effect of zinc acexamate on experimental gastric ulcers: a histochemical study.

    Science.gov (United States)

    Barbarino, F; Toganel, E; Brilinschi, C

    1992-11-01

    The antiulcerogenic effect of zinc acexamate on gastric ulcers induced by reserpine and changes in the morphology of gastric mucosa were studied in rats by histochemical methods. Histochemistry revealed that zinc acexamate preserved reserpine-depleted neutral and acid glycoproteins. ATPase reaction remained strong in nearly normal periglandular capillaries. The reaction intensity of SDH and NADH2-tetrazolium reductase, and the number and size of the DH-positive parietal cells were decreased, illustrating the decline of energy metabolism involved in acid secretion. The decreased height and weaker staining of the pyroninophile chief cell layer corresponded to the lower amount of RNA, an indirect indicator of pepsinogen synthesis. The significant correlation indices "r" between the severity of gastric lesions and histochemical parameters of the defensive (glycoproteins and microvascular ATPase) and aggressive factors (parietal cell DH and chief cell RNA) confirmed the pathogenic effect of reserpine and the protection provided by zinc acexamate. These findings confirm the multifactorial mechanism of action described for zinc acexamate in several previous works.

  4. Histochemical and biometric study of the gastrointestinal system of Hyla orientalis (Bedriaga, 1890) (Anura, Hylidae).

    Science.gov (United States)

    Akat, E; Arıkan, H; Göçmen, B

    2014-11-28

    This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different.

  5. Occurrence, Development and Histochemical Characteristics of Lymphocystis in Cultured Japanese Flounder (Paralichthys olivaceus)

    Institute of Scientific and Technical Information of China (English)

    Sheng Xiuzhen(绳秀珍); Zhan Wenbin

    2004-01-01

    Occurrence and development of lymphocystis cells in flounder Paralichthys olivaceus, and some histochemical features are studied using light and electron microscopy and histochemical methods. Fibroblasts engulf virus particles forming phagocytosis vacuole in the cytoplasm. The infected cells proliferate and turn round and hypertrophied; basophilic inclusion bodies, Feulgen-positive, are present in the cytoplasm, and a hyaline capsule, containing acid ploysaccharide and protein, appears outside cell membrane. The mature lymphocystis cells possess numerous virus particles (200~220nm in diameter with a central core 120~140nm) and high electron-dense inclusions with budding virions at the surface in the cytoplasm, and show margination of chromatin. Moderate electron density particles (70~80nm in diameter) fill out perinuclear cisterna and arrange in crystalloid arrays in nucleus, liberated into the cytoplasm by rupture of nuclear membrane. No virus particles are observed in nucleus. Therefore, inclusion bodies are the sites of virus assembly and maturity, while replication processes of lymphocystis virus are involved in both nuclear and cytoplasmic phases. Besides, strongly basophilic link-like structures are present between adjacent cells or through connective tissue, and virus particles are observed in somewhere of the capsule, thus where may be the spread route of virus from one cell to another or to the environment. The senile lymphocystis cells release virus by the rupture of cells.

  6. Detection of native starches in meat products using histochemical Lugol Calleja method

    Directory of Open Access Journals (Sweden)

    Matej Pospiech

    2014-02-01

    Full Text Available Starch has been still used in food industry today as one of the main additives in foodstuffs. The reason for the use of starches in foodstuffs is their ability to bind water and to contribute to the coherent structure of the final product. However, the presence of starch in some foodstuffs is limited by legislation. These are especially meat products where legislation prohibits using starches. This study deals with determination of native starches using histochemical Lugol Calleja staining in meat products. The targeted structures of this successive staining are starches and collagen ligaments. Other structures can also be detected, based on the knowledge of their morphology. Within the scope of this study, the possibility of histochemical proof on the basis of reaction between Lugol's iodine solution and starch amylose was demonstrated. From the samples analyzed, the following criteria for the method were determined: Repeatability and repeatability of intralaboratory results was 100%, selectivity was determined to be 1.03, specificity of the method was determined to be 0.9, limit of detection was established to be 100% for 0.001 g.kg-1 of the addition, and 87.7% for the concentration of 0.0001 g.kg-1 of the starch addition. Based on the results it is obvious that the method is suitable for determination of native starches in meat products, and, in combination with staining of other foodstuffs ingredients, it gives a complex view of the composition and structure of the meat product.

  7. Electrophysiological, histochemical, and hormonal adaptation of rat muscle after prolonged hindlimb suspension

    Science.gov (United States)

    Kourtidou-Papadeli, Chrysoula; Kyparos, Antonios; Albani, Maria; Frossinis, Athanasios; Papadelis, Christos L.; Bamidis, Panagiotis; Vivas, Ana; Guiba-Tziampiri, Olympia

    2004-05-01

    The perspective of long-duration flights for future exploration, imply more research in the field of human adaptation. Previous studies in rat muscles hindlimb suspension (HLS), indicated muscle atrophy and a change of fibre composition from slow-to-fast twitch types. However, the contractile responses to long-term unloading is still unclear. Fifteen adult Wistar rats were studied in 45 and 70 days of muscle unweighting and soleus (SOL) muscle as well as extensor digitorum longus (EDL) were prepared for electrophysiological recordings (single, twitch, tetanic contraction and fatigue) and histochemical stainings. The loss of muscle mass observed was greater in the soleus muscle. The analysis of electrophysiological properties of both EDL and SOL showed significant main effects of group, of number of unweighting days and fatigue properties. Single contraction for soleus muscle remained unchanged but there was statistically significant difference for tetanic contraction and fatigue. Fatigue index showed a decrease for the control rats, but increase for the HLS rats. According to the histochemical findings there was a shift from oxidative to glycolytic metabolism during HLS. The data suggested that muscles atrophied, but they presented an adaptation pattern, while their endurance in fatigue was decreased.

  8. Classification of the intrafusal muscle fibres in the frog muscle spindle: histochemical and immunofluorescent studies.

    Science.gov (United States)

    Yoshimura, A; Fujitsuka, N; Sokabe, M; Naruse, K; Nomura, K; Diwan, F H; Ito, F

    1990-10-01

    Intrafusal muscle fibres from bull-frog semitendinosus, iliofibularis and sartorius muscles were classified into three types using the histochemical, immunofluorescent and morphological characteristics, with reference to the extrafusal muscle fibres, which were classified into five types in accordance with Rowlerson & Spurway (1988). Immunofluorescent reactions with antibodies against slow or fast myosins obtained from anterior or posterior latissimus dorsi muscles (ALD or PLD), respectively, of chicken were used as the primary criterion. Histochemical profiles of muscle fibres were classified into nine types of myosin ATPase activity as the secondary criterion. Anti-PLD intrafusal fibres (polar zone) with ATPase profiles of moderate to high acid and alkaline stabilities correspond to large nuclear bag fibres in the classification of Diwan & Ito (1989), whereas anti-ALD fibres (polar zone) with alkaline-labile ATPase profiles correspond to medium nuclear bag fibres. On the basis of diameter, anti-PLD fibres (polar zone) with ATPase profiles of moderate to low acid stability and moderate to high alkaline stability seem to correspond to two types of small nuclear chain fibre. Variations between muscles, between intra- and extrafusal fibres and also between zones along intrafusal fibres are discussed.

  9. Morphological, histochemical and immunohistochemical characterization of secretory production of the ciliary glands in the porcine eyelid

    Directory of Open Access Journals (Sweden)

    T Yasui

    2009-06-01

    Full Text Available In addition to performing general histology and cytology of the ciliary glands of the miniature pig, we studied the localization of glycoconjugates and b-defensins in these glands with the use of carbohydrate histochemical and immunohistochemical methods. The secretory cells of the glands were equipped with non-homogeneous secretory granules, a welldeveloped Golgi apparatus and rough endoplasmic reticulum. The secretory epithelium and luminal secretion of the glands contained large amounts of neutral and acidic glycoconjugates with various saccharide residues (a-L-Fuc, b-DGal, a-D-GalNAc and sialic acid. The sebaceous glands and tarsal glands also exhibited positive reactions to most of the histochemical methods. Additionally, the antimicrobial peptide group of b-defensins was demonstrated to be products of the ciliary glands, as well as the sebaceous glands and tarsal glands. The results obtained are discussed with regard to the specific function of the ciliary glandular secretions. These secretory products may be related to the moistening and general protection of the skin surface of the eyelid and ocular surface.

  10. Histochemical characterisation of complex carbohydrates expressed in the alimentary tract of chickens.

    Science.gov (United States)

    Scocco, Paola; Pedini, Vera

    2010-08-01

    This study characterised the composition of surface and secretion complex carbohydrates in the alimentary tract of organically and intensively housed chickens. Histochemical labelling was carried out on samples of oesophagus, glandular stomach and proximal duodenum using (1) alcian blue (AB) at pH 2.5 combined with neuraminidase digestion (Sial) with or without saponification (KOH), (2) AB at pH 1 and 0.5, (3) periodic acid-Schiff (PAS), (4) a combined AB-PAS stain, and (5) both low and high iron diamine stains combined with enzymatic treatments specific for glycosaminoglycans. Semi-quantitative scoring of the histochemical staining indicated abundant acidic sulfated glycoconjugates in each alimentary tract region, in addition to the presence of sialoglycoderivatives in the oesophagus. These complex carbohydrate components are likely to have various functions including the provision of a protective visco-elastic barrier and a role in the defence against specific pathogens. The findings provide a starting point for a more in-depth study of alimentary tract glycoconjugates in poultry.

  11. Cystathionine γ-lyase, a H2S-generating enzyme, is a GPBAR1-regulated gene and contributes to vasodilation caused by secondary bile acids.

    Science.gov (United States)

    Renga, Barbara; Bucci, Mariarosaria; Cipriani, Sabrina; Carino, Adriana; Monti, Maria Chiara; Zampella, Angela; Gargiulo, Antonella; d'Emmanuele di Villa Bianca, Roberta; Distrutti, Eleonora; Fiorucci, Stefano

    2015-07-01

    GPBAR1 is a bile acid-activated receptor (BAR) for secondary bile acids, lithocholic (LCA) and deoxycholic acid (DCA), expressed in the enterohepatic tissues and in the vasculature by endothelial and smooth muscle cells. Despite that bile acids cause vasodilation, it is unclear why these effects involve GPBAR1, and the vascular phenotype of GPBAR1 deficient mice remains poorly defined. Previous studies have suggested a role for nitric oxide (NO) in regulatory activity exerted by GPBAR1 in liver endothelial cells. Hydrogen sulfide (H2S) is a vasodilatory agent generated in endothelial cells by cystathionine-γ-lyase (CSE). Here we demonstrate that GPBAR1 null mice had increased levels of primary and secondary bile acids and impaired vasoconstriction to phenylephrine. In aortic ring preparations, vasodilation caused by chenodeoxycholic acid (CDCA), a weak GPBAR1 ligand and farnesoid-x-receptor agonist (FXR), was iberiotoxin-dependent and GPBAR1-independent. In contrast, vasodilation caused by LCA was GPBAR1 dependent and abrogated by propargyl-glycine, a CSE inhibitor, and by 5β-cholanic acid, a GPBAR1 antagonist, but not by N(5)-(1-iminoethyl)-l-ornithine (l-NIO), an endothelial NO synthase inhibitor, or iberiotoxin, a large-conductance calcium-activated potassium (BKCa) channels antagonist. In venular and aortic endothelial (HUVEC and HAEC) cells GPBAR1 activation increases CSE expression/activity and H2S production. Two cAMP response element binding protein (CREB) sites (CREs) were identified in the CSE promoter. In addition, TLCA stimulates CSE phosphorylation on serine residues. In conclusion we demonstrate that GPBAR1 mediates the vasodilatory activity of LCA and regulates the expression/activity of CSE. Vasodilation caused by CDCA involves BKCa channels. The GPBAR1/CSE pathway might contribute to endothelial dysfunction and hyperdynamic circulation in liver cirrhosis.

  12. Causes of death due to hematological and non-hematological cancers in 57 US patients with type 1 Gaucher Disease who were never treated with enzyme replacement therapy.

    Science.gov (United States)

    Weinreb, Neal J; Lee, Robert E

    2013-01-01

    Patients with type 1 Gaucher disease (GD1) have increased risk of developing myeloma, other hematological cancers, hepatocellular carcinoma, and other solid tumors. Patient awareness of the GD1-cancer association causes anxiety and fear. Little is known about cancer as a cause of death in GD1, especially in patients never treated with GD1-specific therapies. Consequently, the effect of treatment on cancer mortality in GD1 patients is difficult to evaluate. In this review, starting with a population of 184 GD1 cases never treated, we annotate and analyze the causes of death of 57 GD1 patients who died of cancer. The proportional mortality ratio (PMR) for all malignancies in patients with GD1 is 1.57 (p = 0.0002), but it is much higher for myeloma (PMR = 9.66) and other hematological cancers, hepatocellular carcinoma, and kidney cancer (PMR = ≍4). However, deaths from colorectal and pancreatic cancers were not more frequent than expected, and deaths from lung, breast, gynecological, and prostate cancer occurred less than anticipated. Herein, we discuss whether GD1 is truly a hereditary cancer syndrome and the problem of comorbidities and cancer risk assessment, and we speculate as to whether the variability in death by cancer type might be attributable to biochemical sequelae of tumor cell and macrophage/stromal cell GBA1 mutation affecting signals for metastasis, the process most closely associated with cancer mortality.

  13. [Effect of a strong constant magnetic field and a hypomagnetic surrounding medium on histochemical indices of albino rat liver].

    Science.gov (United States)

    Shust, I V; Kostinik, I M

    1975-01-01

    The paper presents histochemical measurements of the liver of white rats exposed to strong and weak constant magnetic fields. A repeated influence (30 min a day) of the strong magnetic field of 912 ersted for 16 days inhibits synthetic processes in the liver tissue. A more prolonged (30 days) exposure to the field produces insignificant histochemical changes which seem to be induced by the occuring compensatory reaction. It is shown that a month long stay of animals in the environment shielded from the geomagnetic field may enhance synthetic processes in some cells and inhibit them in others.

  14. An angiotensin I-converting enzyme mutation (Y465D causes a dramatic increase in blood ACE via accelerated ACE shedding.

    Directory of Open Access Journals (Sweden)

    Sergei M Danilov

    Full Text Available BACKGROUND: Angiotensin I-converting enzyme (ACE metabolizes a range of peptidic substrates and plays a key role in blood pressure regulation and vascular remodeling. Thus, elevated ACE levels may be associated with an increased risk for different cardiovascular or respiratory diseases. Previously, a striking familial elevation in blood ACE was explained by mutations in the ACE juxtamembrane region that enhanced the cleavage-secretion process. Recently, we found a family whose affected members had a 6-fold increase in blood ACE and a Tyr465Asp (Y465D substitution, distal to the stalk region, in the N domain of ACE. METHODOLOGY/PRINCIPAL FINDINGS: HEK and CHO cells expressing mutant (Tyr465Asp ACE demonstrate a 3- and 8-fold increase, respectively, in the rate of ACE shedding compared to wild-type ACE. Conformational fingerprinting of mutant ACE demonstrated dramatic changes in ACE conformation in several different epitopes of ACE. Cell ELISA carried out on CHO-ACE cells also demonstrated significant changes in local ACE conformation, particularly proximal to the stalk region. However, the cleavage site of the mutant ACE--between Arg1203 and Ser1204--was the same as that of WT ACE. The Y465D substitution is localized in the interface of the N-domain dimer (from the crystal structure and abolishes a hydrogen bond between Tyr465 in one monomer and Asp462 in another. CONCLUSIONS/SIGNIFICANCE: The Y465D substitution results in dramatic increase in the rate of ACE shedding and is associated with significant local conformational changes in ACE. These changes could result in increased ACE dimerization and accessibility of the stalk region or the entire sACE, thus increasing the rate of cleavage by the putative ACE secretase (sheddase.

  15. Bioequivalence studies of tibolone in premenopausal women and effects on expression of the tibolone-metabolizing enzyme AKR1C (aldo-keto reductase) family caused by estradiol.

    Science.gov (United States)

    Kang, Keon W; Kim, Yoon G

    2008-12-01

    This study aimed to investigate the bioequivalence of a test formulation of tibolone with the marketed reference formulation in 24 young healthy female volunteers. Tibolone is a synthetic steroid hormone for menopausal women. Volunteers were treated with the 2 formulations of tibolone (total dose of active ingredient 2.5 mg) according to a 2 x 2 crossover design with a 1-week washout period. Plasma concentrations of 3alpha- and 3beta-hydroxytibolone, which are major metabolites of tibolone, were assayed in timed samples over a 24-hour period with a validated gas chromatography/mass spectrometry (GC/MS) method that had a lower limit of quantification of 0.5 ng/mL. The reference and test formulations gave a mean 3alpha-hydroxytibolone C(max) of 5.0 and 5.2 ng/mL, respectively, and a mean 3beta-hydroxytibolone C(max) of 16.4 and 16.5 ng/mL, respectively. The mean AUC(t) of 3alpha-hydroxytibolone was 24.7 and 24.3 ng h/mL, whereas the mean AUC(t) of 3beta-hydroxytibolone was 57.6 and 54.8 ng h/mL for the test and reference formulations, respectively. The authors did not find significant differences in pharmacokinetic parameters between the 2 formulations, but metabolite formation was different from reports in postmenopausal women. The authors therefore measured the effects of estradiol on the expression of the tibolone-metabolizing enzymes, from the aldo-keto reductase (AKR1C) family, using HepG2 cell (human hepatoma cells) and MCF-7 cell (human breast cancer cells). Estradiol increased mRNA levels of AKR1C1, AKR1C2, and AKR1C3 and protein levels of total AKR1C in HepG2 cells. Estradiol selectively enhanced levels of AKR1C2 mRNA in MCF-7 cells. Thus, changes in the major metabolites of tibolone might result from changes in AKR1C family expression by patient estrogen status.

  16. Gene structure and mutations of glutaryl-coenzyme A dehydrogenase: Impaired association of enzyme subunits that is due to an A421V substitution causes glutaric acidemia type I in the Amish

    Energy Technology Data Exchange (ETDEWEB)

    Biery, B.J.; Stein, D.E.; Goodman, S.I. [Univ. of Colorado School of Medicine, Denver, CO (United States)] [and others

    1996-11-01

    The structure of the human glutaryl coenzyme A dehydrogenase (GCD) gene was determined to contain 11 exons and to span {approximately}7 kb. Fibroblast DNA from 64 unrelated glutaric academia type I (GA1) patients was screened for mutations by PCR amplification and analysis of SSCP. Fragments with altered electrophoretic mobility were subcloned and sequenced to detect mutations that caused GA1. This report describes the structure of the GCD gene, as well as point mutations and polymorphisms found in 7 of its 11 exons. Several mutations were found in more than one patient, but no one prevalent mutation was detected in the general population. As expected from pedigree analysis, a single mutant allele causes GA1 in the Old Order Amish of Lancaster County, Pennsylvania. Several mutations have been expressed in Escherichia coli, and all produce diminished enzyme activity. Reduced activity in GCD encoded by the A421V mutation in the Amish may be due to impaired association of enzyme subunits. 13 refs., 5 figs., 3 tabs.

  17. Morphological, histochemical, and interstitial pressure changes in the tibialis anterior muscle before and after aortofemoral bypass in patients with peripheral arterial occlusive disease

    Directory of Open Access Journals (Sweden)

    Grigoriadis Nikolaos

    2002-02-01

    Full Text Available Abstract Background Morphological and electrophysiological studies of ischemic muscles in peripheral arterial disease disclosed evidence of denervation and fibre atrophy. The purpose of the present study is to describe morphological changes in ischemic muscles before and after reperfusion surgery in patients with peripheral occlusive arterial disease, and to provide an insight into the effect of reperfusion on the histochemistry of the reperfused muscle. Methods Muscle biopsies were obtained from the tibialis anterior of 9 patients with chronic peripheral arterial occlusive disease of the lower extremities, before and after aortofemoral bypass, in order to evaluate the extent and type of muscle fibre changes during ischemia and after revascularization. Fibre type content and muscle fibre areas were quantified using standard histological and histochemical methods and morphometric analysis. Each patient underwent concentric needle electromyography, nerve conduction velocity studies, and interstitial pressure measurements. Results Preoperatively all patients showed muscle fibre atrophy of both types, type II fibre area being more affected. The mean fibre cross sectional area of type I was 3,745 μm2 and of type II 4,654 μm2 . Fibre-type grouping, great variation in fibre size and angular fibres were indicative of chronic dennervation-reinnervation, in the absence of any clinical evidence of a neuropathic process. Seven days after the reperfusion the areas of both fibre types were even more reduced, being 3,086 μm2 for type I and 4,009 μm2 for type II, the proportion of type I fibres, and the interstitial pressure of tibialis anterior were increased. Conclusions The findings suggest that chronic ischemia of the leg muscles causes compensatory histochemical changes in muscle fibres resulting from muscle hypoxia, and chronic dennervation-reinnervation changes, resulting possibly from ischemic neuropathy. Reperfusion seems to bring the oxidative

  18. [STUDY OF GLUTATHIONE AND ENZYMES OF ITS METABOLISM UNDER THE INFLUENCE OF ENRICHED DAIRY PRODUCTS FOR PROPHYLACTIC USE IN CONDITIONS OF OXIDATIVE STRESS CAUSED BY EXCESSIVE EXERCISE].

    Science.gov (United States)

    Boyarskaya, L A; Turchaninov, D V; Efremenko, E S; Bogdashin, I V; Vilms, E A; Yunatskaya, T A

    2015-01-01

    There was performed the study of the activity of the antioxidant system of glutathione under regular taking of enriched with micronutrients dairy products "Bifidin " and "Prolacta" in conditions ofsimulation of xcessive exercise loads. Target of research: white outbred male rats weighing 180-200 g (n = 68). Research methods and equipment: information retrieval, biochemical, and statistical. In the subacute experiment there was established that in conditions of excessive exercise loads (CEEL) the body of the rat meets with significant oxidative stress, causing the suppression of the activity of glutathione system: the number of reduced glutathione and the activity of selenium-dependent glutathione-peroxidase decline. Regular taking of dairy products "Bifidin" and "Prolacta" in conditions of CEEL (active training process) allows effectively to neutralize lipid peroxidation processes and normalize indices of the system of glutathione. There is established an increase in the activity ofglutathione-S-transferase in the homogenates of liver tissue in the simulation CEEL in combination with the use ofproduct "Bifidin".

  19. HISTOCHEMICAL AND ELECTRON-MICROSCOPIC CHARACTERIZATION OF HEPATIC MACROPHAGE SUBFRACTIONS ISOLATED FROM NORMAL AND LIPOSOMAL MURAMYL DIPEPTIDE TREATED RATS

    NARCIS (Netherlands)

    HOEDEMAKERS, RMJ; ATMOSOERODJOBRIGGS, JE; MORSELT, HWM; DAEMEN, T; SCHERPHOF, GL; HARDONK, MJ

    1995-01-01

    Subfractions of the hepatic macrophage population, differing in cell size, were isolated from normal rats and rats treated with liposomal muramyl dipeptide (lipMDP) and analyzed histochemically and by ultrastructural peroxidase cytochemistry. The majority of cells in all subfractions of control rats

  20. Muscles within muscles: a tensiomyographic and histochemical analysis of the normal human vastus medialis longus and vastus medialis obliquus muscles.

    Science.gov (United States)

    Travnik, Ludvik; Djordjevič, Srdjan; Rozman, Sergej; Hribernik, Marija; Dahmane, Raja

    2013-06-01

    The aim of this study was to show the connection between structure (anatomical and histochemical) and function (muscle contraction properties) of vastus medialis obliquus (VMO) and vastus medialis longus (VML). The non-invasive tensiomyography (TMG) method was used to determine the contractile properties (contraction time; T(c)) of VML and VMO muscle, as a reflection of the ratio between the slow and fast fibers in two groups of nine young men. VML and VMO significantly (P knee, and to the VMO, which maintains the stable position of the patella in the femoral groove. Our results obtained by TMG provided additional evidence that muscle fibers within the segments of VM muscle were not homogenous with regard to their contractile properties, thereby confirming the histochemical results. T(c) can be attributed to the higher percentage of slow-twitch fibers - type 1. The statistically shorter T(c) (P ≤ 0.001) of VMO (22.8 ± 4.0 ms) compared with VML (26.7 ± 4.0 ms) in our study is consistent with previously found differences in histochemical, morphological and electrophysiological data. In conclusion, the results of this study provide evidence that the VML and VMO muscles are not only anatomically and histochemically different muscles, but also functionally different biological structures.

  1. Chemical composition, quality and histochemical characteristics of individual dromedary camel (Camelus dromedarius) muscles.

    Science.gov (United States)

    Kadim, I T; Al-Karousi, A; Mahgoub, O; Al-Marzooqi, W; Khalaf, S K; Al-Maqbali, R S; Al-Sinani, S S H; Raiymbek, G

    2013-03-01

    This study characterized the chemical composition, quality and histological traits of six muscles from 10 dromedary carcasses. There were significant differences in moisture, fat, protein, mineral, saturated and unsaturated fatty acid contents between muscles. The longissimus thoracis (LT) had the highest cooking loss (33.5%) and triceps brachii (TB) the lowest (29.2%). The shear force value of semitendinosus (ST), semimembranosus (SM) and biceps femoris (BF) were significantly higher than infraspinatus (IS), TB and LT. The LT had significantly higher values for L*, a*, b* than ST. The SM had the lowest MFI (65.3), while IS had the highest value (75.8). The ST significantly had the highest and lowest proportions of Type I and Type IIA muscle fibers, respectively than other muscles. This study indicated that composition, quality, and histochemical parameters varied among camel muscles and the knowledge of this variation allows for better marketing and processing of camel meat.

  2. [A case of MELAS associated with histochemical findings of muscles characteristic of MERRF].

    Science.gov (United States)

    Suzuki, Hidefumi; Ono, Michio; Kojima, Yasuhiro; Kanda, Masutaro; Shibasaki, Hiroshi; Oki, Mituaki; Nakano, Satoshi

    2016-01-01

    We here report a 39-year-old woman of short stature with sensorineural deafness, who suddenly developed status epilepticus. T2-weighed image of brain magnetic resonance imaging (MRI) revealed a high signal lesion in the left temporal area, the distribution of which was not compatible with any particular arterial supply. Lactate and pyruvate were elevated in the serum and cerebrospinal fluid. As the mitochondrial gene analysis revealed the m.3243A>G mutation, diagnosis of mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episode (MELAS) was made. In the histochemical study of a biopsied muscle, the intramuscular blood vessels reacted strongly with SDH (SSV), but the SSV was negative for cytochrome c oxidase (COX), the findings characteristic of myoclonic epilepsy with ragged-red fibers (MERRF). This is the first case of MELAS in which the muscle histochemistry showed positive SSV unassociated with increased COX.

  3. Histochemical Investigation and Kinds of Alkaloids in Leaves of Different Developmental Stages in Thymus quinquecostatus

    Directory of Open Access Journals (Sweden)

    Haiting Jing

    2014-01-01

    Full Text Available Thymus quinquecostatus, with more medical value, is a kind of wild plants. In order to exploit and utilize this plant, we studied the species and locations of alkaloids in its leaves. In this paper, histochemical study of leaves at different developing stages was taken to localize the alkaloids. Meanwhile, the kinds and content of alkaloids in leaves were identified using GC-MS technique. It was found that there were two kinds of glandular trichomes, namely, peltate trichomes and capitate trichomes, on the surface of leaves, and their secretory cells could secrete alkaloids. Results showed that trichomes could secrete alkaloids as soon as the first pair of leaves formed, and there were altogether 18 kinds of alkaloids identified by GC-MS. Nearly all of these alkaloids of leaves at different developing stages were distinct from each other, except one, 3-methoxy-a-methyl-benzeneethanamine, persists at different developing stages with high concentration.

  4. Histochemical analysis of glycoconjugates in the eccrine glands of the raccoon digital pads

    Directory of Open Access Journals (Sweden)

    T Yasui

    2009-06-01

    Full Text Available The distribution and selectivity of complex carbohydrates in the eccrine glands of the digital pads in the North American raccoon (Procyon lotor were studied using light and electron microscopic histochemical methods, particularly lectin histochemistry. In the eccrine glands, the dark cells exhibited neutral and acidic glycoconjugates with different saccharide residues (a-L-fucose, b-D-galactose, b-N-acetyl-D-glucosamine and N-acetyl-neuraminic acid; the clear cells contained numerous glycogen particles and showed a distinct reaction of a-L-fucose. The presence of complex carbohydrates with various terminal sugars was evident in the excretory duct cells. In addition, b-D-galactose and N-acetylneuraminic acid residues were mainly observed in the luminal secretion. The glycoconjugates produced by the eccrine glands of the raccoon digital pads may protect the epidermis against physical damage or microbial contamination. In this way, the normal functioning of the sensory apparatus of the foot pads is ensured.

  5. [Development and differentiation of the rat epididymis. II: histochemical aspects of the peritubular zone].

    Science.gov (United States)

    Francavilla, F; Santiemma, V; Francavilla, S; Forcella, G; Moscardelli, S; Properzi, G

    1979-07-15

    Development of the contractile peritubular structures of rat testis, from birth to full maturation, was investigated by histochemical evaluation of alkaline phosphatase activity (A.P.A.) at 0, 16, 22 e 35 days of age. A.P.A. appeared at 6 days, age of appearance also of the cytoplasmic microfilaments of peritubular cells. The correlation between cytoplasmic microfilaments and A.P.A. was confirmed by the pattern of A.P.A. positivity at 6 degree day, when the inner peritubular cells layer, in which the microfilaments are present, are more positive than the outer layer in which the microfilaments are not yet present. These findings give further support to the belief that the cytoplasmic microfilaments of peritubular cells are contractile structures.

  6. Histological and Histochemical Analyses of the Cuttlebone Sac of the Golden Cuttlefish Sepia esculenta

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The secretion function of mantle is closely related to shell formation in some bivalves and gastropods. Up to now, few researches have been reported for cuttlebone formation in the class Cephalopoda. In this study, the structure and secretion function of cuttlebone sac of the golden cuttlefish Sepia esculenta was analyzed using the histological and histochemical methods. The results showed that high and columnar cells located in sac epithelium, and flat cells existed near the base membrane. A lot of fibroblasts were found in the lateral mantle collective tissue. Some mucus, mucopolysaccharide and alkaline phosphatase (ALP) were found in the sac. The ultrastructural characteristics of Quasi-connective-tissue-calcium cells (QCTCC) were observed using a transmission electron microscope (TEM). The relationship between cuttlebone sac secretion function and shell formation was discussed.

  7. Histochemical demonstration of zinc in rat epididymis using a sulphide-silver method.

    Science.gov (United States)

    Fujimori, O; Tsukise, A; Yamada, K

    1988-01-01

    We studied the histochemical distribution of zinc in rat epididymis using a sulphide-silver method. In the supranuclear cytoplasm of the principal cells that line the epididymis of rats, varying amounts of sulphide-silver-reactive zinc were visualized. In adult mating rats, significant amounts of zinc were found in the proximal portion of the epididymis, whereas in non-mating, mature and immature young rats, this heavy metal was most prominent in the distal portion of this organ. In all of the rats studied, zinc was sparsely distributed in the intermediate portion of the epididymis. From these results, it can be assumed that the zinc present in the epithelial lining of rat epididymis plays an important role in the maturation of spermatozoa. The present results represent a useful contribution to our understanding of the functional morphology of rat epididymis.

  8. Histochemical and ultrastructural study of an elastofibroma dorsi coexisting with a high grade spindle cell sarcoma

    Directory of Open Access Journals (Sweden)

    M Alberghini

    2009-06-01

    Full Text Available Elastofibroma dorsi is a pseudotumoral fibroproliferative lesion characterized by polymorphic fiber-like deposits of elastinophilic material. Several theories have been reported explaining the pathogenesis of elastofibroma. Recent cytogenetic studies have demonstrated chromosomal instability in elastofibromas, not normally observed in non-neoplastic tissues. These chromosomal defects are commonly observed in aggressive fibromatosis too. Such clinical observations suggest a multistage pathogenetic mechanism for the onset of elastofibroma. This study, using histochemical, immunohistochemical staining techniques, and ultrastructural examination, describes the detection of an otherwise typical elastofibroma contextual to a high grade sarcoma. Hence, the coexistence of elastofibroma and high-grade sarcoma may suggest a causal link between the two pathological entities. The results obtained suggest that the coexistence of the two pathological entities is conceivably coincidental.

  9. Histochemical Differential Diagnosis and Polarization Optical Analysis of Amyloid and Amyloidosis

    Directory of Open Access Journals (Sweden)

    M. Bély

    2006-01-01

    Full Text Available Amyloidosis is characterized by extracellular deposition of protein fibrils of chemically heterogeneous composition. Early recognition and identification of amyloid deposits allows an early start of therapy, which may entail a better prognosis. Congo red staining according to Romhányi (1971 is a highly specific and sensitive method for early microscopic recognition of amyloidosis. The main and most important types of amyloidosis may be distinguished by classic histochemical methods of performate pretreatment according to Romhányi (1979, or by KMnO4 oxidation according to Wright (1977 followed by Congo red staining and viewed under polarized light. Differences in the speed of breakdown (disintegration of amyloid deposits according to Bély and Apáthy allow a more precise distinction of various types of amyloid.

  10. Histomorphological and mucin histochemical study of the alimentary canal of pangas catfish, Pangasius pangasius (Hamilton 1822

    Directory of Open Access Journals (Sweden)

    Javd Sadeghinezhad

    2017-06-01

    Full Text Available The present study describes the histological and mucin histochemical properties of the alimentary canal (AC of the pangas catfish, Pangasius pangasius. The results revealed that the mucosa of the oesophagus was lined by a stratified epithelium containing chloride cells and taste buds which suggested mechanic, gustatory and physiologic roles of the oesophagus in this species. The stomach mucosa was lined by a simple columnar epithelium. The lamina propria-submucosa in cardiac and fundic stomach contained gastric glands. The pyloric stomach had the thickest muscularis layer among all the parts of the AC. The villi showed the maximum height and width in the middle intestine. The tunica muscularis and serosa showed the thinnest thickness among all parts of AC. The mucin histochemistry showed that the goblet cells of oesophagus and intestine contained both neutral and acidic with carboxylated and sulfated mucins and there was not acidic mucins in epithelial cells of the stomach.

  11. Histochemical Structure of Stomach (Proventriculus and Gizzard in Some Bird Species

    Directory of Open Access Journals (Sweden)

    Emel DEMİRBAĞ

    2015-09-01

    Full Text Available Stomach samples (proventriculus and gizzard of sparrow hawk (Accipiter nisus, crow (Corvus corone and sparrow (Passer domesticus were investigated in this study. In histochemical investigations, strong sulfated mucins were not determined in compound glands of proventriculus, but a little reaction to AB (pH 0.5 in compound glands was observed in proventriculus of sparrow hawk and crow. While weak AB (pH 1.0 (+ character was found in sparrow gizzard, it was found as dense in superficial glands of sparrow hawk. However, it was not observed in crow gizzard. While AF (+ character (sulfated mucins was detected as dense in epithelium and as weak in glands of gizzard of sparrow hawk, this character was not observed in glands of gizzards of crow and sparrow. Any reactivity to Periodic Acid-Shiff (PAS staining was not observed in both epithelium and glands of sparrow proventriculus

  12. Histochemical distribution of zinc in the brain of the rainbow trout, Oncorhynchos myciss. I. The telencephalon

    DEFF Research Database (Denmark)

    Piñuela, C; Baatrup, E; Geneser, F A

    1992-01-01

    The present paper which describes the distribution of zinc in the telencephalon of the rainbow trout, Oncorhynchos myciss, is the first report on the distribution of a heavy metal in the fish brain. Zinc was demonstrated histochemically by silver enhancement using the Neo-Timm method. The staining...... of cytoarchitectural features. However, the telencephalon stained much more weakly than the rest of the brain, in striking contrast to the situation in the reptilian, mammalian, and avian brain. In these classes, high staining intensities are observed almost exclusively in the telencephalon. The staining...... was essentially restricted to the nuclei of the ventral telencephalic area. In the dorsal telencephalic area, only the medial and central zones and medial part of the posterior zone showed comparable staining intensities. The Neo-Timm staining pattern lends support to the view that the pallio-subpallial boundary...

  13. Histological and histochemical characterization of the secretory cells of Choeradoplana iheringi Graff, 1899 (Platyhelminthes: Tricladida: Terricola

    Directory of Open Access Journals (Sweden)

    S. A. de Souza

    Full Text Available The present study aims at providing a detailed description of the histology, as well as the first histochemical characterization, of the secretory cells of the epidermis, pharynx, and copulatory organs of Choeradoplana iheringi, in order to give further support to studies on the physiology of these organs. The secretory cells are distinguished on the basis of secretion morphology and its staining properties, using trichrome methods and histochemical reactions. Four cell types open through the epidermis of Ch. iheringi, three of them secreting basic protein and a fourth containing glycosaminoglycan mucins. The epidermal lining cells store glycogen. In the pharynx, four secretory cell types were distinguished. Two types produce glycoprotein, a third type secretes basic protein, and another one produces glycosaminoglycan mucins. In the male copulatory organs, the prostatic vesicle receives four secretory cell types containing basic protein, except for one type which produces glycoprotein. The two secretory cell types opening into the male atrium secrete, respectively, glycoprotein, and glycosaminoglycan mucins. In the female copulatory organs, the female atrium and its proximal diverticulum, the vagina, receive two types of secretory cells producing, respectively, basic protein and glycosaminoglycan mucins. Another secretory cell type constitutes the so-called shell glands which open into the common glandular duct, secreting basic protein. The lining cells of the male and female atria produce a mucous secretion containing glycosaminoglycans. In addition, the lining epithelium of the female atrium presents an apical secretion of a proteic nature. The occurrence of a kind of spermatophore is reported for the first time for a species of Choeradoplana. This structure is located in the male or female atria in different specimens, and characterized by erythrophil, xanthophil, and/or mixed secretions associated with sperm.

  14. Acrylamide alters glycogen content and enzyme activities in the liver of juvenile rat.

    Science.gov (United States)

    Kovac, Renata; Rajkovic, Vesna; Koledin, Ivana; Matavulj, Milica

    2015-10-01

    Acrylamide (AA) is spontaneously formed in carbohydrate-rich food during high-temperature processing. It is neurotoxic and potentially cancer causing chemical. Its harmful effects on the liver, especially in a young organism, are still to be elucidated. The study aimed to examine main liver histology, its glycogen content and enzyme activities in juvenile rats treated with 25 or 50mg/kg bw of AA for 3 weeks. Liver samples were fixed in formalin, routinely processed for paraffin embedding, sectioning and histochemical staining. Examination of haematoxylin and eosin (H&E)-stained sections showed an increase in the volume of hepatocytes, their nuclei and cytoplasm in both AA-treated groups compared to the control. In Periodic acid-Schiff (PAS)-stained sections in low-dose group was noticed glycogen reduction, while in high-dose group was present its accumulation compared to the control, respectively. Serum analysis showed increased activity of aspartate aminotransferase (AST), and decreased activity of alanine aminotransferase (ALT) in both AA-treated groups, while the activity of alkaline phosphatase (ALP) was increased in low-dose, but decreased in high-dose group compared to the control, respectively. Present results suggest a prominent hepatotoxic potential of AA which might alter the microstructural features and functional status in hepatocytes of immature liver.

  15. Effect of angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers on all-cause mortality, cardiovascular deaths, and cardiovascular events in patients with diabetes mellitus: a meta-analysis.

    Science.gov (United States)

    Cheng, Jun; Zhang, Wen; Zhang, Xiaohui; Han, Fei; Li, Xiayu; He, Xuelin; Li, Qun; Chen, Jianghua

    2014-05-01

    Angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs) may have different effects on cardiovascular (CV) events in patients with diabetes mellitus (DM). To conduct a meta-analysis to separately evaluate the effects of ACEIs and ARBs on all-cause mortality, CV deaths, and major CV events in patients with DM. DATA SOURCES Data sources included MEDLINE (1966-2012), EMBASE (1988-2012), the Cochrane Central Register of Controlled Trials, conference proceedings, and article reference lists. We included randomized clinical trials reporting the effects of ACEI and ARB regimens for DM on all-cause mortality, CV deaths, and major CV events with an observation period of at least 12 months. Studies were excluded if they were crossover trials. Dichotomous outcome data from individual trials were analyzed using the risk ratio (RR) measure and its 95% CI with random-effects models. We estimated the difference between the estimates of the subgroups according to tests for interaction. We performed meta-regression analyses to identify sources of heterogeneity. Primary end points were all-cause mortality and death from CV causes. Secondary end points were the effects of ACEIs and ARBs on major CV events. Twenty-three of 35 identified trials compared ACEIs with placebo or active drugs (32,827 patients) and 13 compared ARBs with no therapy (controls) (23,867 patients). When compared with controls (placebo/active treatment), ACEIs significantly reduced the risk of all-cause mortality by 13% (RR, 0.87; 95% CI, 0.78-0.98), CV deaths by 17% (0.83; 0.70-0.99), and major CV events by 14% (0.86; 0.77-0.95), including myocardial infarction by 21% (0.79; 0.65-0.95) and heart failure by 19% (0.81; 0.71-0.93). Treatment with ARBs did not significantly affect all-cause mortality (RR, 0.94; 95% CI, 0.82-1.08), CV death rate (1.21; 0.81-1.80), and major CV events (0.94; 0.85-1.01) with the exception of heart failure (0.70; 0.59-0.82). Both ACEIs and ARBs were

  16. Histochemical and immunohistochemical analyses of the myocardial scar fallowing acute myocardial infarction

    Directory of Open Access Journals (Sweden)

    Tatić Vujadin

    2012-01-01

    Full Text Available Background/Aim. The heart has traditionally been considered as a static organ without capacity of regeneration after trauma. Currently, the more and more often asked question is whether the heart has any intrinsic capacities to regenerate myocytes after myocardial infarction. The aim of this study was to present the existence of the preserved muscle fibers in the myocardial scar following myocardial infarction as well as the presence of numerous cells of various size and form that differently reacted to the used immunohistochemical antibodies. Methods. Histological, histochemical and immunohistochemical analyses of myocardial sections taken from 177 patients who had died of acute myocardial infarction and had the myocardial scar following myocardial infarction, were carried out. More sections taken both from the site of acute infarction and scar were examined by the following methods: hematoxylin-eosin (HE, periodic acid schiff (PAS, PAS-diastasis, Masson trichrom, Malory, van Gieson, vimentin, desmin, myosin, myoglobin, alpha actin, smoth muscle actin (SMA, p53, leukocyte common antigen (LCA, proliferating cell nuclear antigen (PCNA, Ki-67, actin HHF35, CD34, CD31, CD45, CD45Ro, CD8, CD20. Results. In all sections taken from the scar region, larger or smaller islets of the preserved muscle fibers with the signs of hypertrophy were found. In the scar, a large number of cells of various size and form: spindle, oval, elongated with abundant cytoplasm, small with one nucleus and cells with scanty cytoplasm, were found. The present cells differently reacted to histochemical and immunohistochemical methods. Large oval cells showed negative reaction to lymphocytic and leukocytic markers, and positive to alpha actin, actin HHF35, Ki-67, myosin, myoglobin and desmin. Elongated cells were also positive to those markers. Small mononuclear cells showed positive reaction to lymphocytic markers. Endothelial and smooth muscle cells in the blood vessel walls

  17. Histological and histochemical analysis of the gastrointestinal tract of the common pipistrelle bat (Pipistrellus pipistrellus).

    Science.gov (United States)

    Strobel, S; Encarnação, J A; Becker, N I; Trenczek, T E

    2015-01-01

    Bats have a very high mass-specific energy demand due to small size and active flight. European bat species are mostly insectivorous and the morphology of the gastrointestinal tract should be adapted accordingly. This study investigated the general anatomy by histology and the function by analysing carbohydrate distribution in particular of the mucus of the GI tract of the insectivorous bat Pipistrellus pipistrellus. The GI tracts of three individuals were dissected, fixed in formaldehyde, and embedded in paraffin wax. The tissues and cells of the GI tract of P. pipistrellus were analysed by classical (Acid Alizarin Blue, Haematoxylin-Eosin, and Masson Goldner Trichrome), histochemical (periodic acid-Schiff, Alcian blue at pH 2.5) and lectin histochemical (lectins WGA and HPA) staining procedures. The GI tract of P. pipistrellus was organised into the typical mammalian layers. The short, narrow, and thin-walled esophagus was simple with a folded stratified squamous epithelium without glands but mucous surface cells secreting neutral mucus. The stomach was globular shaped without specialisation. Mucous surface cells produced neutral mucus whereas neck and parietal cells secreted a mixture of neutral and acid mucus. Chief cell surface was positive for N-acetylglucosamine and the cytoplasm for N-acetylgalactosamine residues. The intestine lacked a caecum and appendix. The small intestine was divided into duodenum, jejunum‑ileum and ileum‑colon. The epithelium consisted of columnar enterocytes and goblet cells. The large intestine was short, only represented by the descending colon-rectum. It lacked villi and the mucosa had only crypts of Lieberkühn. Towards the colon-rectum, goblet cells produced mucus with N-acetylglucosamine residues increasing in acidity except in colon-rectum where acidity was highest in the base of crypts. Along the tube the surface of enterocytes was positive for N-acetylglucosamine and N-acetylgalactosamine. All over the mucus filling

  18. Histological and histochemical analysis of the gastrointestinal tract of the common pipistrelle bat (Pipistrellus pipistrellus

    Directory of Open Access Journals (Sweden)

    S. Strobel

    2015-04-01

    Full Text Available Bats have a very high mass-specific energy demand due to small size and active flight. European bat species are mostly insectivorous and the morphology of the gastrointestinal tract should be adapted accordingly. This study investigated the general anatomy by histology and the function by analysing carbohydrate distribution in particular of the mucus of the GI tract of the insectivorous bat Pipistrellus pipistrellus. The GI tracts of three individuals were dissected, fixed in formaldehyde, and embedded in paraffin wax. The tissues and cells of the GI tract of P. pipistrellus were analysed by classical (Acid Alizarin Blue, Haematoxylin-Eosin, and Masson Goldner Trichrome, histochemical (periodic acid-Schiff, Alcian blue at pH 2.5 and lectin histochemical (lectins WGA and HPA staining procedures. The GI tract of P. pipistrellus was organised into the typical mammalian layers. The short, narrow, and thin-walled esophagus was simple with a folded stratified squamous epithelium without glands but mucous surface cells secreting neutral mucus. The stomach was globular shaped without specialisation. Mucous surface cells produced neutral mucus whereas neck and parietal cells secreted a mixture of neutral and acid mucus. Chief cell surface was positive for N-acetylglucosamine and the cytoplasm for N-acetylgalactosamine residues. The intestine lacked a caecum and appendix. The small intestine was divided into duodenum, jejunum‑ileum and ileum‑colon. The epithelium consisted of columnar enterocytes and goblet cells. The large intestine was short, only represented by the descending colon-rectum. It lacked villi and the mucosa had only crypts of Lieberkühn. Towards the colon-rectum, goblet cells produced mucus with N-acetylglucosamine residues increasing in acidity except in colon-rectum where acidity was highest in the base of crypts. Along the tube the surface of enterocytes was positive for N-acetylglucosamine and N-acetylgalactosamine. All over the

  19. Food Enzymes

    Science.gov (United States)

    McBroom, Rachel; Oliver-Hoyo, Maria T.

    2007-01-01

    Many students view biology and chemistry as two unrelated, separate sciences; how these courses are generally taught in high schools may do little to change that impression. The study of enzymes provide a great opportunity for both biology and chemistry teachers to share with students the interdisciplinary nature of science. This article describes…

  20. Food Enzymes

    Science.gov (United States)

    McBroom, Rachel; Oliver-Hoyo, Maria T.

    2007-01-01

    Many students view biology and chemistry as two unrelated, separate sciences; how these courses are generally taught in high schools may do little to change that impression. The study of enzymes provide a great opportunity for both biology and chemistry teachers to share with students the interdisciplinary nature of science. This article describes…

  1. Enzyme immunoassay

    DEFF Research Database (Denmark)

    Feldt-Rasmussen, B; Dinesen, B; Deckert, M

    1985-01-01

    An enzyme linked immunoadsorbent assay for urinary albumin using commercially available reagents is described. The assay range is 2.5-120 micrograms/l. When samples are analysed in two standard dilutions, the assayable albumin concentration range is 2.5-240 mg/l, covering the clinical range from...

  2. Identifying motor and sensory myelinated axons in rabbit peripheral nerves by histochemical staining for carbonic anhydrase and cholinesterase activities

    Science.gov (United States)

    Riley, Danny A.; Sanger, James R.; Matloub, Hani S.; Yousif, N. John; Bain, James L. W.

    1988-01-01

    Carbonic anhydrase (CA) and cholinesterase (CE) histochemical staining of rabbit spinal nerve roots and dorsal root ganglia demonstrated that among the reactive myeliated axons, with minor exceptions, sensory axons were CA positive and CE negative whereas motor axons were CA negative and CE positive. The high specificity was achieved by adjusting reaction conditions to stain subpopulations of myelinated axons selectively while leaving 50 percent or so unstained. Fixation with glutaraldehyde appeared necessary for achieving selectivity. Following sciatic nerve transection, the reciprocal staining pattern persisted in damaged axons and their regenerating processes which formed neuromas within the proximal nerve stump. Within the neuromas, CA-stained sensory processes were elaborated earlier and in greater numbers than CE-stained regenerating motor processes. The present results indicate that histochemical axon typing can be exploited to reveal heterogeneous responses of motor and sensory axons to injury.

  3. Mucin Reactivity after Acute Exposure to Mercury and Zinc Chloride in Neurocytes of Freshwater Snail Bellamya bengalensis: A Histochemical Study

    Directory of Open Access Journals (Sweden)

    Sunil Londhe

    2014-01-01

    Full Text Available Central nervous system (CNS of gastropods is extensively studied for its pathology and immunocytology; there is scanty information on histochemical alterations in neuronal cells due to heavy metal. This study was designed to investigate histochemical alterations in cerebral neurons of freshwater snail Bellamya bengalensis after intoxication with mercuric chloride (HgCl2 and zinc chloride (ZnCl2. As per chemical components cerebral ganglia of Bellamya bengalensis showed three different zones, namely, periganglionic connective tissue sheath, interperikaryonal space, and neuropil. After intoxication, increased acidic content in extracellular matrix (ECM of neuronal cells was found. These alterations were directly proportional to time of exposure period. From results, HgCl2 proved its highly toxic nature over chemical composition of cerebral neuronal cells of freshwater snail Bellamya bengalensis compared to ZnCl2 intoxication.

  4. Sperm enzyme activity analysis of individual sperm for detection of heritable mutations in mammals.

    Science.gov (United States)

    Panda, B B; Ficsor, G; Ginsberg, L C

    1988-03-15

    Male mice were injected i.p. with 2.5 mg/kg mitomycin C, 100 mg/kg ethyl nitrosourea or saline and mated with untreated virgin females five weeks later. Sperm from 64 of the F1 male progeny were analyzed histochemically for acrosin, succinic dehydrogenase and alpha-glycerophosphate dehydrogenase activity. The frequency of F1 males with sub-normal sperm enzyme activity was significantly higher among progeny from treated males than in controls. These results show that analysis of sperm enzyme activity in F1 males is a practical method for detection of transmitted mutations induced in a treated parent.

  5. Microscopic analysis and histochemical observations of the medicinal root of Iostephane heterophylla (Cav.) Benth. ex Hemsl. (Asteraceae)

    OpenAIRE

    Estela Sandoval; Robert A. Bye; Griselda Ríos; María Isabel Aguilar

    2005-01-01

    The roots of Iostephane heterophylla are popular in Mexican traditional medicine and as such are a good candidate to develop herbal drug preparations to be used as phytomedicine. International criteria for validation and standardization of a herbal product as phytomedicine include, among others, the integration of microscopic and histochemical characteristics of the raw material, as in this case the herbal drug, to guarantee its authenticity. As an original contribution to the knowledge of th...

  6. [Histochemical detection of glycoproteins and glycosaminoglycans in the respiratory mucosa of albino rats during estrous cycle, pregnancy and puerperium].

    Science.gov (United States)

    Pontes, P A; Simões, M J; Merzel, J

    1989-11-01

    In this work we attempted to detect, with histochemical methods, the possible modifications in the mucus of the respiratory mucosa of albino female rats during estral cycle, pregnancy and puerperium. Based on its results, it was possible to conclude that: a--There were no modifications in the nature of the epithelial and supraepithelial mucus during the studied periods: b--The Alcian Blue staining from lamina propria is absent during pregnancy and present during puerperium.

  7. Atypical micromorphology and uncommon location of cryptococcosis: a histopathologic study using special histochemical techniques (one case report).

    Science.gov (United States)

    Gazzoni, Alexandra Flávia; Severo, Cecília Bittencourt; Barra, Marines Bizarro; Severo, Luiz Carlos

    2009-04-01

    Here we report an unusual case of disseminated cryptococcosis in a patient with AIDS. Although typical Cryptococcus neoformans micromorphology was observed in tongue biopsy, cervical lymph node examination revealed atypical histopathologic findings. These included pseudohyphae, chains of budding yeasts and structures resembling germ tubes. Cryptococcus neoformans infection in supraclavicular lymph nodes was also confirmed by culture. The importance of using special histochemical techniques-Mayer's mucicarmine stain for mucicarminophilic capsule and Grocott's silver stain-in the diagnosis of cryptococcosis is reinforced.

  8. Biochemical and Histochemical Localization of Monoterpene Biosynthesis in the Glandular Trichomes of Spearmint (Mentha spicata) 12

    Science.gov (United States)

    Gershenzon, Jonathan; Maffei, Massimo; Croteau, Rodney

    1989-01-01

    The primary monoterpene accumulated in the glandular trichomes of spearmint (Mentha spicata) is the ketone (−)-carvone which is formed by cyclization of the C10 isoprenoid intermediate geranyl pyrophosphate to the olefin (−)-limonene, hydroxylation to (−)-trans-carveol and subsequent dehydrogenation. Selective extraction of the contents of the glandular trichomes indicated that essentially all of the cyclase and hydroxylase activities resided in these structures, whereas only about 30% of the carveol dehydrogenase was located here with the remainder located in the rest of the leaf. This distribution of carveol dehydrogenase activity was confirmed by histochemical methods. Electrophoretic analysis of the partially purified carveol dehydrogenase from extracts of both the glands and the leaves following gland removal indicated the presence of a unique carveol dehydrogenase species in the glandular trichomes, suggesting that the other dehydrogenase found throughout the leaf probably utilizes carveol only as an adventitious substrate. These results demonstrate that carvone biosynthesis takes place exclusively in the glandular trichomes in which this natural product accumulates. Images Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 PMID:16666709

  9. Biochemical and histochemical localization of monoterpene biosynthesis in the glandular trichomes of spearmint (Mentha spicata)

    Energy Technology Data Exchange (ETDEWEB)

    Gershenzon, J.; Maffei, M.; Croteau, R. (Washington State Univ., Pullman (USA))

    1989-04-01

    The primary monoterpene accumulated in the glandular trichomes of spearmint (Mentha spicata) is the ketone (-)-carvone which is formed by cyclization of the C{sub 10} isoprenoid intermediate geranyl pyrophosphate to the olefin (-)-limonene, hydroxylation to (-)-trans-carveol and subsequent dehydrogenation. Selective extraction of the contents of the glandular trichomes indicated that essentially all of the cyclase and hydroxylase activities resided in these structures, whereas only about 30% of the carveol dehydrogenase was located here with the remainder located in the rest of the leaf. This distribution of carveol dehydrogenase activity was confirmed by histochemical methods. Electrophoretic analysis of the partially purified carveol dehydrogenase from extracts of both the glands and the leaves following gland removal indicated the presence of a unique carveol dehydrogenase species in the glandular trichomes, suggesting that the other dehydrogenase found throughout the leaf probably utilizes carveol only as an adventitious substrate. These results demonstrate that carvone biosynthesis takes place exclusively in the glandular trichomes in which this natural product accumulates.

  10. Histochemical changes during the ontogeny of malagueta and biquinho pepper seeds

    Directory of Open Access Journals (Sweden)

    Haynna Fernandes Abud

    2017-08-01

    Full Text Available This study aimed to evaluate the structural changes and reserve deposition of malagueta and biquinho pepper seeds and to determine their chemical composition during ontogeny. The fruits were collected 25, 40, 55, 70, 85, and 100 days after anthesis (DAA. After collection, the seeds were extracted and fixed in FAA 50 and then stored in 70% ethanol. Subsequently, the seeds were embedded in methacrylate (Historesin and sectioned with a microtome. The sections were stained with Toluidine blue O for anatomical analysis and Xylidine Ponceau, Periodic acid Schiff (PAS and Lugol’s iodine solution for histochemical analysis. Sections were made using a cryomicrotome for acid phloroglucinol staining and Sudan III tests. Progressive lignification of the seed coat cells was observed. Proteins and lipids were the main reserve compounds of malagueta and biquinho pepper seeds stored in the embryo and endosperm. Starch served as a transient reserve during embryo development. The lipid reserves were observed in embryo and endosperm cells from 25 until 100 DAA. At 100 DAA, the protein contents of the malagueta and biquinho pepper seeds were 0.33 and 0.62 g g-1 DM and the lipid contents were 0.30 and 0.25 g g-1 DM, respectively.

  11. Histochemical and structural analysis of mucous glycoprotein secreted by the gill of Mytilus edulis

    Energy Technology Data Exchange (ETDEWEB)

    Ahn, Hae-Young.

    1988-01-01

    Studies were carried out to characterized various mucous cells in the gill filament, to ascertain structural characteristics of the secreted mucous glycoproteins, and to determine the ability of the gill epithelium to incorporate ({sup 14}C)glucosamine as a precursor in the biosynthesis and secretion of mucous glycoproteins. Using histochemical staining techniques, mucous cells containing neutral and acidic mucins were found in the lateral region, whereas mucous cells containing primarily neutral or sulfated mucins were found in the postlateral region. Serotonin, but not dopamine, stimulated the mucous secretion. In tissues pretreated with ({sup 14}C)glucosamine, the secreted glycoproteins contain incorporated radiolabel. Analysis by column chromatography using Bio-Gel P-2 and P-6 shows that the secretion contains two glycoprotein populations. Glycoprotein II has a molecular weight of 2.3 {times} 10{sup 4} daltons. Upon alkaline reductive borohydride cleavage of the O-glycosidic linkages of glycoprotein I, about 70% of the radiolabel was removed from the protein. Gas chromatographic analysis of the carbohydrate composition shows that the glycoproteins contains N-acetylglucosamine (GluNAc), N-acetylgalactosamine (GalNAc), and galactose, fucose and mannose. Amino acid analysis shows that the glycoproteins are rich in serine, threonine and proline.

  12. The epithelium of the tongue of Ambystoma mexicanum. Ultrastructural and histochemical aspects.

    Science.gov (United States)

    Wistuba, J; Opolka, A; Clemen, G

    1999-12-01

    The distribution pattern of taste buds and goblet cells and histochemical and ultrastructural aspects of the tongue epithelium of Ambystoma mexicanum are here described. This study is also concerned with the developmental stages and origins of the epithelial cells. Pavement cells and goblet cells of the stratum superficiale are replaced by basal cells of the stratum germinativum in larvae and neotenous adults. The pavement cells of the larvae are characterized by a marginal layer of mucin grana. Decompaction of the mucins occurs immediately before extrusion in the adult. The larval goblet cell type (type I), which is also present in the adult, contains unfused grana of irregular shape. At the tip of the tongue, a further type (type II) of goblet cells is found. In the type II cells the intracellular secretory grana fuse to a single homogeneous mass. Leydig cells of the tongue epithelium are discerned by light microscopy first in the semi-adult, apparently correlated with partial metamorphosis. In the course of ontogenesis and induced metamorphosis the secretion changes to neutral glycoconjugates. The mucins of the pavement cells change first followed by those of the goblet cells. The glands of the secondary tongue show a dorso-ventral pattern of varying secretory qualities. Taste buds are found at the anterior margin of the tongue and along the base of the gill clasps in the early larva. They are already distributed all over the tongue at the end of the early larval phase.

  13. Histological and histochemical aspects of the penial glands of Girardia biapertura Sluys, 1997 (Platyhelminthes, Tricladida, Paludicola

    Directory of Open Access Journals (Sweden)

    S. T. SOUZA

    Full Text Available Girardia biapertura was described with sperm ducts penetrating the penis bulb, subsequently opening separately at the tip of the penis papilla and receiving the abundant secretion of penial glands. In the present work, the penial glands of this species have been histologically and histochemically analysed, and four types of secretory cells are distinguished. The openings of the penial glands into the intrabulbar and intrapapillar sperm ducts, designated here as intrapenial ducts, allow for the distinction between three histologically differentiated regions. The most proximal region possibly corresponds to the bulbar cavity of other freshwater triclads whereas the median and distal portions correspond to the ejaculatory duct. The proximal region of the intrapenial ducts receives mainly the openings of a secretory cell type (type I that produces a proteinaceous secretion. A second type of secretory cell (type II that secretes neutral mucopolyssacharides opens into the median region of the intrapenial ducts. The distal portion of the ducts receives two types of secretory cells (types III and IV which secret glycoprotein and glycosaminoglycans, respectively. Types III and IV open also directly into the male atrium through the epithelium of the penis papilla. A comparison with the results presented here and those of other authors for species of Girardia is provided and the importance of the study of the penial glands for taxonomic characterisation of freshwater triclads is emphasised.

  14. Organogenesis of the digestive tract in the white seabream, Diplodus sargus. Histological and histochemical approaches.

    Science.gov (United States)

    Ortiz-Delgado, J B; Darias, M J; Cañavate, J P; Yúfera, M; Sarasquete, C

    2003-10-01

    The ontogeny of the digestive tract of the white seabream, Diplodus sargus during the larval development up to day 45 post-hatching (dph) has been studied using histological and histochemical techniques. The oesophageal goblet cells appeared around 6 dph and contained neutral and acid mucosubstances (PAS/diastase-PAS and Alcian Blue pH 2.5 positive reactions). An incipient stomach can be distinguished from 2 dph but the first sign of gastric gland development was detected around 13-15 dph, increasing in number and size by 22-23 dph. Gastric glands were concentrated in the cardiac stomach region and they had a high content of protein rich in tyrosine, arginine and tryptophan. Acidophilic supranuclear inclusions related to pynocitosis of proteins, were already observed in the intestinal cells of the posterior intestine around 4-6 dph (exogenous feeding) and they were present until 25 dph. The intestinal mucous cells appeared between 15-18 dph and contained a mixture of neutral and acid mucosubstances/glycoconjugates, carboxylated ones being more abundant than the sulphated ones. The stomach and gastric glands were fully developed by the first month of life marking the beginning of digestive features characteristic of the juvenile stage. Around 4-6 dph, glycogen, proteins and neutral lipids were observed in the granular cytoplasm of hepatocytes. Strongly acidophilic zymogen granules were also present, at this time, in the basophilic cytoplasm of the exocrine pancreatic acinar cells and contained abundant proteins, especially rich in arginine, tyrosine and tryptophan.

  15. Histochemical and Immunohistochemical Analysis of the Stomach of Rhinella icterica (Anura, Bufonidae

    Directory of Open Access Journals (Sweden)

    Clarice Machado-Santos

    2014-01-01

    Full Text Available The stomach of Rhinella icterica was analyzed at light microscopy, employing histochemical techniques, lectin histochemistry, and immunohistochemistry for identifying enteroendocrine cells (EC. Although the stomach was composed of fundic and pyloric regions, its wall is formed by mucosa, submucosa, muscularis, and serosa. The mucosa was lined by a simple columnar mucous epithelium, supported by loose connective tissue. Several tubular, simple glands were composed of mucous neck cells, containing oxynticopeptic cells and EC cells. The mucous neck cells were rich in neutral glycoconjugates. The oxynticopeptic cells were predominant in fundic glands, exhibiting weaker alcianophilic reaction at their apical cytoplasm. Serotonin (5-HT immunoreactive (IR cells occurred throughout the entire stomach, preferentially located among mucous cells at upper part of the fundic glands. The muscularis mucosae, formed of smooth muscle, separated the mucosal layer from the submucosa, both of which were constituted by loose connective tissue, but without glands. Lymphoid modules occurred in the mucosa at the boundary at the stomach and the gut. In addition, the muscularis was constituted by two sublayers, the circular internal and the longitudinal external, being recovered by the connective tissue of the serosa.

  16. Morphological, histological and histochemical analysis of the digestive tract of Trachelyopterusstriatulus (Siluriformes: Auchenipteridae

    Directory of Open Access Journals (Sweden)

    Marcella L. dos Santos

    2015-08-01

    Full Text Available ABSTRACT The digestive tract of Trachelyopterus striatulus (Steindachner, 1877 was studied using morphological, histological, and histochemical techniques. The barbels, lips and tongue had stratified squamous epithelium with mucous, claviform cells, and taste buds. Trachelyopterus striatulus had a dental plaque with villiform teeth. The bucopharyngeal cavity was formed by the gill apparatus and pharyngeal teeth with vilifform denticles. The oesophagus presented stratified squamous epithelium with mucous cells and taste buds. The stomach included cardiac, fundic, and pyloric regions and a simple prismatic epithelium with prismatic cells that reacted positively to periodic acid-Schiff (PAS and amylase+PAS. Only the cardiac and pyloric regions responded positively to alcian blue pH 2.5 (Ab pH 2.5 and alcian blue pH 0.5 (Ab pH 0.5. The cardiac and fundic regions exhibited tubular gastric glands. The intestine was 118.90 ± 22.49 mm long with an intestinal coefficient (CO of 0.83 ± 0.13. The epithelium was simple prismatic with a brush border and goblet cells, and a greater number of goblet cells were found in the caudal region of the intestine. The mucous cells and goblet cells reacted positively to PAS, amylase+PAS, Ab pH 2.5, and Ab pH 0.5. We analysed if the function of the mucosubstances and morphological characteristics of the digestive tract of T. striatulus are compatible with omnivorous feeding habit.

  17. Histochemical characterization of early response to Cochliobolus sativus infection in selected barley genotypes.

    Science.gov (United States)

    Rodríguez-Decuadro, Susana; Silva, Paula; Bentancur, Oscar; Gamba, Fernanda; Pritsch, Clara

    2014-07-01

    Much effort is being made to breed barley with durable resistance to leaf spot blotch incited by Bipolaris sorokiniana (teleomorph: Cochliobolus sativus). We hypothesized that susceptibility and resistance traits in 11 diverse barley genotypes inoculated with a single C. sativus isolate might specify a range of distinct host cell responses. Quantitative descriptions of interaction microphenotypes exhibited by different barley genotype seedlings after infection with C. sativus are provided. Early oxidative responses occurring in epidermis and mesophyll leaf tissue were monitored by histochemical analysis of H2O2 accumulation at 8, 24, and 48 h after inoculation. Cell wall apposition (CWA) in epidermal cells and hypersensitive reaction (HR) of epidermal or mesophyll tissue were early defenses in both resistant and susceptible genotypes. There were differences in level, duration, and frequency of occurrence for CWA and HR for the different barley genotypes. Occurrence of HR in epidermal cells at post-penetration stages was indicative of compatibility. Patterns of cell responses were microphenotypically diverse between different resistant and susceptible genotypes. This suggests that timing and level of response are key features of microphenotypic diversity that distinguish different functional mechanisms of resistance and susceptibility present in barley.

  18. Needle muscle biopsy: technique validation and histological and histochemical methods for evaluating canine skeletal muscles

    Directory of Open Access Journals (Sweden)

    Sérgio de Almeida Braga

    2017-05-01

    Full Text Available This study evaluated the needle muscle biopsy technique using a 6G Bergström percutaneous needle combined with histological and histochemical methods to analyze the skeletal muscle of dogs. There are few studies about canine skeletal muscles and a lack of reports in the literature about tissue collection and analysis for canine species. Evaluation of 32 German Shepherd samples collected from the gluteus medius, at a depth of 3 cm, was performed. The choice of gluteus medius and the 3-cm depth provided good quantity fragments with sufficient sizes (3–5 mm, which permitted optimal visualization of muscle fibers. Myosin ATPase, at pH 9.4, 4.6, and 4.3, and SDH reactions revealed that all muscle samples analyzed had fibers in the classic mosaic arrangement, enabling counting and typification. The mean percentages of fibers were 29.95% for type I and 70.05% for type II. On the basis of these results, we concluded that the percutaneous needle biopsy technique for canine skeletal muscles is a safe and easy procedure that obtains fragments of proper sizes, thereby enabling the study of muscle fibers. Standardization of the muscle of choice and the depth of muscle sample collection significantly contributed to this success. This is an important method to evaluate muscle fiber types of dogs and diagnose important diseases affecting the skeletal muscles.

  19. Histological, surface ultrastructural, and histochemical study of the stomach of red piranha, Pygocentrus nattereri (Kner

    Directory of Open Access Journals (Sweden)

    Ghosh Saroj Kumar

    2015-12-01

    Full Text Available The structural characterization and function of the stomach in the omnivore Pygocentrus nattereri were described using light and scanning electron microscopy. The sac-like stomach was morphologically divided into the cardiac and pyloric regions. The histological structure of the stomach consisted of four layers of the mucosa, submucosa, muscularis, and serosa. The superficial epithelium of the cardiac stomach was lined with columnar epithelial cells and the glandular epithelium contained numerous gastric glands. Gastric glands were completely absent in the pyloric portion. The mucosal surface of the stomach was a meshwork of various folds, provided with oval or rounded columnar epithelial cells which were densely packed with short, stubby microvilli. The occasional presence of conspicuous gastric pits was surrounded by epithelial cells. The localization and chemical nature of acid and neutral mucins in the various cells of the stomach was studied by employing combined the Alcian Blue-Periodic Acid Schiff (AB-PAS technique. The deposition of glycogen was detected in the gastric glands as well as in the epithelial lining of the stomach. The utmost reactions for protein and tryptophan were recorded in the gastric glands of mucosa. The cellular organization and histochemical characterization of the stomach are discussed in relation to the feeding and digestion of the fish concerned.

  20. Ultrastructural and histochemical properties of the olfactory system in the japanese jungle crow, Corvus macrorhynchos.

    Science.gov (United States)

    Kondoh, Daisuke; Nashimoto, Mai; Kanayama, Shunsaku; Nakamuta, Nobuaki; Taniguchi, Kazuyuki

    2011-08-01

    Although it has been commonly believed that birds are more dependent on the vision and audition than the olfaction, recent studies indicate that the olfaction of birds is related to the reproductive, homing, and predatory behaviors. In an attempt to reveal the dependence on the olfactory system in crows, we examined the olfactory system of the Japanese jungle crow (Corvus macrorhynchos) by histological, ultrastructural, and lectin histochemical methods. The olfactory epithelium (OE) of the crow occupied remarkably a small area of the nasal cavity (NC) and had the histological and ultrastructural features like other birds. The olfactory bulb (OB) of the crow was remarkably small and did not possess the olfactory ventricle. The left and right halves of the OB were fused in many cases. In the lectin histochemistry, soybean agglutinin (SBA) and Vicia villosa agglutinin (VVA) stained a small number of the receptor cells (RCs) in the OE and the olfactory nerve layer (ONL) and glomerular layer (GL) on the dorsocaudal region of the OB. Phaseolus vulgaris agglutinin-E (PHA-E) stained several RCs in the OE and the ONL and GL on the ventral region of the OB. These results suggest that 1) the crow has less-developed olfactory system than other birds, and 2) the dedicated olfactory receptor cells project their axons to the specific regions of the OB in the crow.

  1. The salivary glands of Ameiva ameiva (Teiidae, Lacertilia). A morphological, morphometric and histochemical study.

    Science.gov (United States)

    Lopes, R A; Costa, J R; Piccolo, A M; Petenusci, S O

    1982-01-01

    The authors studied morphological, morphometric, and histochemically the mucosubstances and proteins in the salivary glands of the lizard Ameiva. Based on the results, the authors concluded: 1. The labial salivary gland is formed by small mucous and mucoserous glands; the sublingual gland by mucoserous cells. 2. Mucous cells show neutral and sulphated mucosubstances and sialic acid. Mucoserous cells of the labial gland show neutral mucosubstance, sialic acid, hyaluronic acid and protein radicals. Mucoserous cells of the sublingual gland show neutral mucosubstance, sialic acid and protein radicals. 3. The average values for acinar area were: 1,198.11 microns 2 for mucoserous acini and 2,105.95 microns 2 for mucous acini of the labial salivary gland. The average values for nucleus volume were: 47.41 microns 3 for mucoserous cells and 38.97 microns 4 for mucous cells. 4. The average values for acinar area and nuclear volume of the mucoserous cells of the subingual gland were respectively: 1,474.62 microns 2 and 67.77 microns 3.

  2. Locus ceruleus neurons in people with autism contain no histochemically-detectable mercury.

    Science.gov (United States)

    Pamphlett, Roger; Kum Jew, Stephen

    2016-02-01

    Exposure to environmental mercury has been proposed to play a part in autism. Mercury is selectively taken up by the human locus ceruleus, a region of the brain that has been implicated in autism. We therefore looked for the presence of mercury in the locus ceruleus of people who had autism, using the histochemical technique of autometallography which can detect nanogram amounts of mercury in tissues. In addition, we sought evidence of damage to locus ceruleus neurons in autism by immunostaining for hyperphosphorylated tau. No mercury was found in any neurons of the locus ceruleus of 6 individuals with autism (5 male, 1 female, age range 16-48 years). Mercury was present in locus ceruleus neurons in 7 of 11 (64%) age-matched control individuals who did not have autism, which is significantly more than in individuals with autism. No increase in numbers of locus ceruleus neurons containing hyperphosphorylated tau was detected in people with autism. In conclusion, most people with autism have not been exposed early in life to quantities of mercury large enough to be found later in adult locus ceruleus neurons. Human locus ceruleus neurons are sensitive indicators of mercury exposure, and mercury appears to remain in these neurons indefinitely, so these findings do not support the hypothesis that mercury neurotoxicity plays a role in autism.

  3. Histochemical responses of rats exercised in two weekly frequencies and ingesting standard or hypercaloric diet

    Directory of Open Access Journals (Sweden)

    FI Freitas

    2009-07-01

    Full Text Available This study investigated if overfed rats present morphological and histochemical muscle adaptation similar to normally fed, both submitted to two different weekly frequencies of training. Thirty male Wistar rats were fed either with standard chow (SCØ or with hypercaloric diet (HCØ. They were subdivided into six subgroups: sedentary (SCØ and HCØ, trained twice/week (SC2 and HC2 and trained five times/week (SC5 and HC5. The trained groups swam 60 min/day, during 10 weeks. Twenty four hours after the last training, samples of Gastrocnemius were excised and stained with HE, NADH-TR and m-ATPase, and the capillary density was calculated. Total heart mass (HM and the mass of atrium (AM, left (LV and right (RV ventricles were excised and weighted. The comparisons were made by ANOVA and by Covariance analysis, adjusting the variables by body weight. The results showed that the HCØ achieved higher BM, however, absolute HM did not differ post training. Irrespective of the diet, rats that were trained twice a week presented significantly greater increase in the AM. In general, the SC5 and HC5 groups showed higher HM, LV, RV, proportion of oxidative fibres and capillary density, compared to the sedentary and twice week trained groups. A higher proportion of injuries (splitting was noted in the HC2 and HC5 compared to SC2 and SC5. These results indicate that the frequency of training influenced the skeletal and heart adaptation and larger changes were observed in the 5x/week group, which ingested the standard diet. The 5x/week training groups also presented large amount of muscle fibres damage.

  4. Tracheobronchial epithelium of the sheep: IV. Lectin histochemical characterization of secretory epithelial cells.

    Science.gov (United States)

    Mariassy, A T; Plopper, C G; St George, J A; Wilson, D W

    1988-09-01

    Conventional histochemical characterization of the mucus secretory apparatus is often difficult to reconcile with the biochemical analysis of respiratory secretions. This study was designed to examine the secretory glycoconjugates in airways using lectins with biochemically defined affinities for main sugar residues of mucus. We used five biotinylated lectins--DBA (Dolichos biflorus) and SBA (Glycine max) for N-acetyl galactosamine (galNAc), BSA I (Bandeiraea simplicifolia) and PNA (Arachis hypogea) for galactose (gal), and UEA I (Ulex europeus)--for detection of fucose (fuc) in HgCl2-fixed, paraffin-embedded, serially sectioned trachea, lobar and segmental bronchi and bronchioles of nine sheep. Lectins selectively localized the carbohydrate residues in luminal secretions, on epithelial cell surfaces, and in secretory cells. In proximal airways, the major carbohydrate residues in luminal secretions, cell surfaces, goblet cells, and glands were fuc and gal-NAc. PNA reacted mainly with apical granules of less than 10% of goblet cells, and gal residues were only detected in some of the mucous cells and on basolateral cell surfaces. Distal airways contained sparse secretion in the lumen, mucous cells contained weakly reactive fuc and gal-NAc, and the epithelial surfaces of Clara cells contained gal. Sugars abundant in the airway secretions were also the major component of cells in glands. We conclude that there is a correlation between specific sugar residues in secretory cells, glycocalyx, and luminal secretions in proximal and distal airways. This suggests that lectins may be used to obtain information about airway secretory cell composition from respiratory secretions.

  5. The histochemical assessment of sulpho-, sialo-, and neutral-mucosubstances in fetal gastric mucosa

    Directory of Open Access Journals (Sweden)

    Lokadolalu Chandracharya Prasanna

    2015-01-01

    Full Text Available Background: Mucins are complex composition of carbohydrates and may be present as a mixture of different types. Normal distribution of mucin and its alteration in various inflammatory, benign and malignant lesions of gastrointestinal tract has aroused interest in the field of histochemistry. The main purpose of present work is to study the staining pattern and distribution of cells in different parts of fetal gastric mucosa and to correlate the nature of gastric mucins and its functional significance. Methods: A total of 25 fetus stomach specimens (total 75 samples one sample each from different parts of the stomach like fundus, body and pylorus, from fresh specimens. The samples were washed in normal saline, fixed in 2% calcium acetate in 10% formalin. These tissues were routinely processed and paraffin blocks were prepared. 6 and #61549; sections of these blocks were taken for histological and different histochemical staining. Results: Fetal fundic part of stomach shows increased neutral mucin in surface epithelium and foveolar cells. With combined AB pH 2.5 - PAS technique increased neutral mucin and small amount of acid mucins are observed. With AB pH 1, surface epithelium and deep glands show negative staining. Moderate alcinophilia is observed in deep foveolar cells and glandular cells. AB pH 2.5 shows alcinophilia in surface epithelium, foveolar cells and mucous neck cells indicating presence of sialomucin. Fetal pyloric part of stomach shows increased acid and neutral mucins. With pH 2.5 - PAS staining, purple staining is observed in surface epithelium, deep foveolar and pyloric glands. Conclusion: All types of mucosubstances - neutral, sialo and sulpho-mucins, are secreted in relatively increased amounts by the surface epithelium and the glands of the stomach of the human fetus and neonate. Sulphomucin is seen mainly in the cells of the surface epithelium. [Int J Res Med Sci 2015; 3(1.000: 235-238

  6. Histochemical activity of 5-4-isomerase-3-B hydroxy steroid dehydrogenase in the ovary of the viviparous mexican lizardSceloporus mucronatus (Reptilia:Prhynosomatidae) and interelationship with progesterone levels during pregnancy

    Institute of Scientific and Technical Information of China (English)

    Martn Martnez-torres; E Martha Prez-armendariz; M Elena Hernndez Caballero; Juana luis; guadalupe ortz-Lpez

    2012-01-01

    Objective:To relate the histological characteristics and histochemicalΔ5-4-isomerase-3 beta hydroxy steroid dehydrogenase(Δ5-43β-HSD) activity of the corpora lutea(CL) and the atresic vitellogenic follicles(AVF) with progesterone(P4) plasma concentrations in three different times of gestation (early, medium and late) in the viviparous lizardSceloporus mucronatus (S. mucronatus).Methods:The histological characteristics as well as histochemical activity ofΔ5-43β-HSD of theCL andAVF and their relationship with plasmaP4 levels were studied during three different times of pregnancy of the viviparous lizardS. mucronatus.Results:Corpora lutea develops during the first third of gestation.In second third, the luteal tissue reaches maturity and starts the first regressive changes.The last third of gestation was characterized by a considerably advance in the luteolysis.Activity ofΔ5-43β-HSD was observed in he luteal cell mas.The activity of this enzyme were high during the first third and scantle activity was detected in the last third.Even though atresic vitellogenic follicles are found throughout the whole period of gestation,Δ5-43β-HSD activity is very low in relation with showed byCL and does not change significantly in the studied period of time.Another hand, we observed a direct relationship among the histological aspect of the corpus luteum,Δ5-43β-HSD activity and progesterone levels. Conclusions:These observations suggests that the corpus luteum is the most important source of ovarian progesterone(P4) during pregnancy and that the participation of theAVF in the production of this hormone is little or non-existent.

  7. Somatic mosaicism caused by monoallelic reversion of a mutation in T cells of a patient with ADA-SCID and the effects of enzyme replacement therapy on the revertant phenotype.

    Science.gov (United States)

    Moncada-Vélez, M; Vélez-Ortega, A; Orrego, J; Santisteban, I; Jagadeesh, J; Olivares, M; Olaya, N; Hershfield, M; Candotti, F; Franco, J

    2011-11-01

    Patients with adenosine deaminase (ADA) deficiency exhibit spontaneous and partial clinical remission associated with somatic reversion of inherited mutations. We report a child with severe combined immunodeficiency (T-B- SCID) due to ADA deficiency diagnosed at the age of 1 month, whose lymphocyte counts including CD4+ and CD8+ T and NK cells began to improve after several months with normalization of ADA activity in Peripheral blood lymphocytes (PBL), as a result of somatic mosaicism caused by monoallelic reversion of the causative mutation in the ADA gene. He was not eligible for haematopoietic stem cell transplantation (HSCT) or gene therapy (GT); therefore he was placed on enzyme replacement therapy (ERT) with bovine PEG-ADA. The follow-up of metabolic and immunologic responses to ERT included gradual improvement in ADA activity in erythrocytes and transient expansion of most lymphocyte subsets, followed by gradual stabilization of CD4+ and CD8+ T (with naïve phenotype) and NK cells, and sustained expansion of TCRγδ+ T cells. This was accompanied by the disappearance of the revertant T cells as shown by DNA sequencing from PBL. Although the patient's clinical condition improved marginally, he later developed a germinal cell tumour and eventually died at the age of 67 months from sepsis. This case adds to our current knowledge of spontaneous reversion of mutations in ADA deficiency and shows that the effects of the ERT may vary among these patients, suggesting that it could depend on the cell and type in which the somatic mosaicism is established upon reversion.

  8. Morphological and histochemical characterization of the digestive tract of the puffer fish Sphoeroides testudineus (Linnaeus 1758 (Tetraodontiformes: Tetraodontidae

    Directory of Open Access Journals (Sweden)

    KAINÃ R.C. FAGUNDES

    2016-01-01

    Full Text Available ABSTRACT Morphological analysis of the digestive tract of Sphoeroides testudineus showed an esophagus with an anterior and a posterior portion, the abdominal pouch. No stomach was observed between the abdominal pouch and the intestine. The intestine was arranged in three segments and two loops, and the distal portion had the rectum opening into the anus. Histochemical analyses showed that the esophagus secreted acid mucosecretions, and that there was a qualitative increase in goblet cells from the proximal to distal area of the intestine. The rectum showed cells secreting acid and neutral mucus. Given these features, this species presents a morphology which creates a link between its ecology and behavior.

  9. Histochemical analysis of glycoconjugates in the skin of a catfish (arius tenuispinis, day).

    Science.gov (United States)

    Al-Banaw, A; Kenngott, R; Al-Hassan, J M; Mehana, N; Sinowatz, F

    2010-02-01

    A histochemical study using conventional carbohydrate histochemistry (periodic-acid staining including diastase controls, alcian blue staining at pH 1 and 2.5) as well as using a battery of 14 fluorescein isothiocyanate (FITC)-labelled lectins to identify glycoconjugates present in 10 different areas of the skin of a catfish (Arius tenuispinis) was carried out. The lectins used were: mannose-binding lectins (Con A, LCA and PSA), galactose-binding lectins (PNA, RCA), N-acetylgalactosamine-binding lectins (DBA, SBA, SJA and GSL I), N-acetylglucosamine-binding lectins (WGA and WGAs), fucose-binding lectins (UEA) and lectins which bind to complex carbohydrate configurations (PHA E, PHA L). Conventional glycoconjugate staining (PAS staining, alcian blue at pH 1 and 2.5) showed that the mucous goblet cells contain a considerable amount of glycoconjugates in all locations of the skin, whereas the other unicellular gland type, the club cells, lacked these glycoconjugates. The glycoproteins found in goblet cells are neutral and therefore stain magenta when subjected to PAS staining. Alcian blue staining indicating acid glycoproteins was distinctly positive at pH 1, but gave only a comparable staining at pH 2.5. The mucus of the goblet cells therefore also contains acid glycoproteins rich in sulphate groups. Using FITC-labelled lectins, the carbohydrate composition of the glycoproteins of goblet cells could be more fully characterized. A distinct staining of the mucus of goblet cells was found with the mannose-binding lectins LCA and PSA; the galactosamine-binding lectins DBA, SBA and GLS I; the glucosamine-binding lectin WGA; and PHA E which stains glycoproteins with complex carbohydrate configurations. No reaction occurred with the fucose-binding lectin UEA and the sialic acid-specific lectin SNA. In addition, the galactose-binding lectins PNA and RCA showed only a weak or completely negative staining of the mucus in the goblet cells. The specificity of the lectin staining

  10. The Histochemical Characterization of the Glycoconjugates in the Epidermal Mucous Cells of the Red Californian Earthworm, Eisenia foetida

    Directory of Open Access Journals (Sweden)

    Kenan Çinar

    2014-01-01

    Full Text Available The aim of this study was to characterize the nature and regional distribution of the glycoconjugates secreted by epidermal mucous cells in Eisenia foetida (Annelida. Specimens were divided into six regions from anterior to posterior. The histochemistry was carried out by using standard histochemical methods. Histochemical staining properties of glycoconjugates in epidermal mucous cells were determined regionally. The epidermis of all regions contained strong to stronger PAS (+ cells in various degrees. The epidermis of the first, fourth, fifth, and sixth regions had strong to stronger AB pH 2.5 (+ cells. On the contrary, all regions contained weak to moderate AB pH 0.5 and AB pH 1.0 (+ cells. Most of mucous cells in epidermis of the first region contained both PAS (+ and AB (+ mucosubstances. All regions included weaker to weak AF (+ cells. All regions featured KOH/PAS (+ cells, with a slight reduction in reaction intensity in the epidermis of the last three regions. In this context, the different staining patterns observed in epidermal mucous cells hinted at their functional roles with respect to production of mucus with different physical properties. This study provided comprehensive information about the regional distribution patterns of the glycoconjugates and an opportunity to compare their distributional patterns in other annelids.

  11. Effect of sex on histological and histochemical structures of different parts of the kidney in Japanese quail.

    Science.gov (United States)

    Mobini, Behzad; Abdollahi, MohammadHossein

    2016-09-01

    The aim of the present study was to investigate the effect of gender on the histological and histochemical structures of different anatomical regions of the kidney in Japanese quail (Coturnix japonica). Tissue samples from cranial, middle and caudal divisions of each kidney were obtained from 20 male and 20 female adult, healthy Japanese quail. The sections stained with hematoxylin & eosin ( H & E: ), Masson's trichrome, Verhoeff's, Alcian blue (pH 2.5), Periodic acid-Schiff, and Gomori's method for reticulum. Unusual findings of the kidney in Japanese quail were the presence of three types of nephrons, all the connective tissue fibers in capsule and interlobular septa and AB-reactions of the proximal convoluted cells. No significant sex-based differences were found. The various histological structures of the kidney showed no significant differences among different divisions of the left and right kidneys. It is concluded that the general histological and histochemical properties of the kidney in Japanese quail were similar to those of chickens and some other species, but that there were also some differences. One of the major differences was brush border of interdigitating microvilli on luminal surface of collecting ducts in Japanese quail.

  12. Histochemical detection of creatine kinase in the electrocyte of Electrophorus electricus (L.).

    Science.gov (United States)

    Taffarel, M; Silva, C B; Machado, R D; Hassón-Voloch, A

    1987-01-01

    Detection of creatine kinase, which catalyzes the conversion of ADP and phosphocreatine to ATP and creatine, was performed on the electrocyte of Electrophorus electricus (L.) using a histoenzymological method based on the formation of blue colored formazan. The results indicate that the enzyme is mainly located within the cytoplasm of the electrolyte.

  13. Modulation of glomerular ECTO-ADPase expression by oestradiol - A histochemical study

    NARCIS (Netherlands)

    Faas, MM; Bakker, WW; Klok, PA; Baller, JFW; Schuiling, GA

    1997-01-01

    The effect of 17-beta-oestradiol (OE2) upon the activity of the glomerular anti-thrombotic ecto-enzyme ADPase was studied in cyclic and ovariectomized (OVX) Wistar rats. On day 0 (i.e. at the time of ovariectomy or 11 days after ovariectomy) rats received OE2-releasing Silastic implants or empty imp

  14. Modulation of glomerular ECTO-ADPase expression by oestradiol - A histochemical study

    NARCIS (Netherlands)

    Faas, MM; Bakker, WW; Klok, PA; Baller, JFW; Schuiling, GA

    The effect of 17-beta-oestradiol (OE2) upon the activity of the glomerular anti-thrombotic ecto-enzyme ADPase was studied in cyclic and ovariectomized (OVX) Wistar rats. On day 0 (i.e. at the time of ovariectomy or 11 days after ovariectomy) rats received OE2-releasing Silastic implants or empty

  15. Fibre characteristics and enzyme levels of arm and leg muscles in elite cross-country skiers

    DEFF Research Database (Denmark)

    Mygind, Erik

    1995-01-01

    Performance tests and measurements of maximal aerobic capacity were performed during the competition period in elite cross-country skiers. Muscle biopsies were taken in the middle of January. Histochemical fibre typing, determination of fibre areas and number of capillaries as well as assays....... The FTa fibre area in TRI was significantly larger than in VAS. No differences were found in the number of capillaries per fibre in TRI (2.7) and VAS (2.5). The number of capillaries per area was significantly lower in TRI (373) as compared to VAS (422). The LDHtot enzyme level was significantly higher...

  16. The Use of Cytochrome C Oxidase Enzyme Activity and Immunohistochemistry in Defining Mitochondrial Injury in Kidney Disease.

    Science.gov (United States)

    Zsengellér, Zsuzsanna K; Rosen, Seymour

    2016-09-01

    The renal biopsy is a dynamic way of looking at renal disease, and tubular elements are an important part of this analysis. The mitochondria in 20 renal biopsies were examined by immunohistochemical (electron transport chain enzyme: cytochrome C oxidase IV [COX IV]) and enzyme histochemical methods (COX), both by light and electron microscopy. The distal convoluted tubules and thick ascending limbs showed the greatest intensity in the COX immunostains and enzyme activity in controls. The degree of mitochondrial COX protein and enzyme activity diminished as the tubules became atrophic. With proximal hypertrophic changes, there was great variation in both COX activity and protein expression. In contrast, in three cases of systemic lupus erythematosus, biopsied for high-grade proteinuria, the activity was consistently upregulated, whereas protein expression remained normal. These unexpected findings of heterogeneous upregulation in hypertrophy and the dyssynchrony of protein expression and activity may indicate mitochondrial dysregulation. Functional electron microscopy showed COX activity delineated by the intense mitochondrial staining in normal or hypertrophic proximal tubules. With atrophic changes, residual small mitochondria with diminished activity could be seen. With mitochondrial size abnormalities (enlargement and irregularity, adefovir toxicity), activity persisted. In the renal biopsy, mitochondrial analysis is feasible utilizing immunohistochemical and enzyme histochemical techniques.

  17. Enzyme detection by microfluidics

    DEFF Research Database (Denmark)

    2013-01-01

    Microfluidic-implemented methods of detecting an enzyme, in particular a DNA-modifying enzyme, are provided, as well as methods for detecting a cell, or a microorganism expressing said enzyme. The enzyme is detected by providing a nucleic acid substrate, which is specifically targeted...... by that enzyme...

  18. A Novel Splice-Site Mutation in Angiotensin I-Converting Enzyme (ACE) Gene, c.3691+1G > A (IVS25+1G > A), Causes a Dramatic Increase in Circulating ACE through Deletion of the Transmembrane Anchor

    NARCIS (Netherlands)

    Persu, Alexandre; Lambert, Michel; Deinum, Jaap; Cossu, Marta; de Visscher, Nathalie; Irenge, Leonid; Ambroise, Jerome; Minon, Jean-Marc; Nesterovitch, Andrew B.; Churbanov, Alexander; Popova, Isolda A.; Danilov, Sergei M.; Danser, A. H. Jan; Gala, Jean-Luc

    2013-01-01

    Background: Angiotensin-converting enzyme (ACE) (EC 4.15.1) metabolizes many biologically active peptides and plays a key role in blood pressure regulation and vascular remodeling. Elevated ACE levels are associated with different cardiovascular and respiratory diseases. Methods and Results: Two Bel

  19. Histochemical Study of N-acetylgalactosamine Containing Glycoconjugate in Intraductal Carcinoma of the Breast by HPA Lectin

    Directory of Open Access Journals (Sweden)

    Mohammad Reza Arab

    2006-01-01

    Full Text Available Introduction: Glycoconjugates are a class of glycoproteins or glycolipids, their terminal sugars are responsible for cell-cell and/or cell-extracellular matrix interactions. Aberrant glycosylation of these compounds are one of the most important aspects of cellular transformation, metastasis and escape of tumoral cells from immune system and resistance to antineoplastic drugs. Recent studies showed that patients with HPA (helix pomatia agglutinin positive intraductal carcinoma cells have worse prognosis compared to patients with HPA negative cells. The aim of the present study was to define the presence of GalNac terminal sugar in glycoconjugate of different grades of intraductal breast carcinoma and to compare the degree and the pattern of reactivity of tumoral cells to HPA lectin. Materials and Methods: The paraffin blocks belonging to 20 patients of intraductal carcinoma was chosen from pathology archive of Khatam-Al-Anbia hospital in Zahedan and 5-7 micrometer sections were prepared. Two expert pathologists determined histopathological grading independently. The lectin histochemistry was performed using HPA. The same observers determined histochemical grading. Data were analyzed by NPAR (non-parametric test of Mann Whitney. Results: Results of this study revealed that the pattern and the degree of histochemical reactivity of neoplastic cells differ in all grades of intraductal carcinoma. Histochemical staining showed significant difference between grades of intraductal carcinoma of the breast (p<0.003. The lowest reactivity was seen in grade I and the highest in grade III. Furthermore, the reaction of tumoral cells was primarily confined to apical surfaces of cells in grade I, to the Golgi zone in grade II, and to a diffuse cytoplasmic distribution in grade III. Conclusion: Our data suggest that the HPA reactivity of tumoral cells were different in all grades of intraductal carcinoma. The tumor cells showed aberrant glycosylation, which

  20. Evaluation of micro-energy dispersive X-ray fluorescence and histochemical tests for aluminium detection in plants from High Altitude Rocky Complexes, Southeast Brazil.

    Science.gov (United States)

    Campos, Naiara V; Pereira, Tiago A R; Machado, Mariana F; Guerra, Marcelo B B; Tolentino, Gláucia S; Araújo, Josiane S; Rezende, Maíra Q; Silva, Maria Carolina N A da; Schaefer, Carlos E G R

    2014-03-01

    The soils developed under High Altitude Rocky Complexes in Brazil are generally of very low chemical fertility, with low base saturation and high exchangeable aluminium concentration. This stressful condition imposes evolutionary pressures that lead to ecological success of plant species that are able to tolerate or accumulate high amounts of aluminium. Several analytical methods are currently available for elemental mapping of biological structures, such as micro-X-ray fluorescence (μ-EDX) and histochemical tests. The aim of this study was to combine μ-EDX analysis and histochemical tests to quantify aluminium in plants from High Altitude Rocky Complexes, identifying the main sites for Al-accumulation. Among the studied species, five showed total Al concentration higher than 1000 mg kg-1. The main Al-hyperaccumulator plants, Lavoisiera pectinata, Lycopodium clavatum and Trembleya parviflora presented positive reactions in the histochemical tests using Chrome Azurol and Aluminon. Strong positive correlations were observed between the total Al concentrations and data obtained by μ-EDX analysis. The μ-EDX analysis is a potential tool to map and quantify Al in hyperaccumulator species, and a valuable technique due to its non-destructive capacity. Histochemical tests can be helpful to indicate the accumulation pattern of samples before they are submitted for further μ-EDX scrutiny.

  1. Elevated Liver Enzymes

    Science.gov (United States)

    Symptoms Elevated liver enzymes By Mayo Clinic Staff Elevated liver enzymes may indicate inflammation or damage to cells in the liver. Inflamed or ... than normal amounts of certain chemicals, including liver enzymes, into the bloodstream, which can result in elevated ...

  2. Histochemical Comparison of the Hypopharyngeal Gland in Apis cerana Fabricius, 1793 Workers and Apis mellifera Linnaeus, 1758 Workers

    Directory of Open Access Journals (Sweden)

    Guntima Suwannapong

    2010-01-01

    Full Text Available Hypopharyngeal glands of honeybee are age-dependent structures that change with the size of acini and are correlated with various social behaviors. The histochemical structure of Apis cerana and A. mellifera worker hypopharyngeal glands in four different developmental stages wes stained with ninhydrin Schiff's and periodic acid Schiff's reagents (PAS for localization of proteins and carbohydrates, respectively, and examined with light microscopy. Nurse bees of both honeybee species had significantly larger glands as compared to guards and forgers, but there were no statistically significant differences between these two species after accounting for caste. Gland protein concentration increased progressively in nurse bees, and this was correlated with the appearance of enriched protein granules in the cytoplasm. In addition, the hypopharyngeal gland protein concentration of A. mellifera was higher than that of A. cerana even though gland size was not significantly different between species. However, gland size was shown to have decreased progressively in foragers and guards.

  3. Characterization of atherosclerosis by histochemical and immunohistochemical methods in African grey parrots (Psittacus erithacus) and Amazon parrots (Amazona spp.).

    Science.gov (United States)

    Fricke, Cornelia; Schmidt, Volker; Cramer, Kerstin; Krautwald-Junghanns, Maria-Elisabeth; Dorrestein, Gerry M

    2009-09-01

    The aim of the study was to characterize atherosclerotic changes in African grey parrots (Psittacus erithacus) and Amazon parrots (Amazona spp.) by histochemical and immunohistochemical methods. Samples of the aorta ascendens and trunci brachiocephalici from 62 African grey parrots and 35 Amazon parrots were stained by hematoxylin and eosin and Elastica van Gieson for grading of atherosclerosis in these birds. Four different stages were differentiated. The incidence of atherosclerosis in the examined parrots was 91.9% in African grey parrots and 91.4% in Amazon parrots. To evaluate the pathogenesis in birds, immunohistochemical methods were performed to demonstrate lymphocytes, macrophages, smooth muscle cells, and chondroitin sulfate. According to the missing lymphocytes and macrophages and the absence of invasion and proliferation of smooth muscle cells in each atherosclerotic stage, "response-to-injury hypothesis" seems inapplicable in parrots. Additionally, we found alterations of vitally important organs (heart, lungs) significantly correlated with atherosclerosis of the aorta ascendens.

  4. Histochemical studies on the body wall of nematodes: Haemonchus contortus (Rud., 1803) and Xiphinema insigne Loos, 1949.

    Science.gov (United States)

    Sood, M L; Kalra, S

    1977-04-15

    Histochemical studies on the body wall of Haemonchus contortus (Rud.) and Xiphinema insigne Loos have been made. In H. contortus, the cuticle is mainly proteinous in nature. The lipids and PAS-postive materials are only present in cortical layers. In addition, haemoglobin and acid phosphatase are also present. The hypodermis shows the presence of glycogen, lipids, RNA, acid and alkaline phosphatases. The oval dense body is composed of keratinous and collagenous proteins associated with acid mucopolysaccharides. Muscles carry a greater concentration of glycogen granules and phospholipids. In X. insigne, the cuticle is rich in sudanophilic lipids. The cuticle also consists of weakly acidic mucopolysaccharides. Hypodermis and muscles contain lipids and glycogen. In addition, hypodermis also consists of acidic mucopolysaccharides. The functional significance of these components has been fully discussed.

  5. Histochemical evaluation of catechins in PEG stressed transgenic tea plants using catechin-specific-diazotized sulfanilamide reagent.

    Science.gov (United States)

    Bhattacharya, A; Sharma, M; Gulati, A; Joshi, R; Chanda, S K; Ahuja, P S

    2015-01-01

    We investigated the applicability of catechin-specific-reagent (CSR) for histochemical evaluation of catechins. The diazotized arylamine moiety in CSR reacts specifically with the A-ring of catechins to yield a golden yellow complex. This makes it highly specific for spectrophotometric quantification of catechins. Therefore, microtome cut sections of untransformed and osmotin-expressing transgenic leaves and stem of tea were stained with CSR. We found catechins in the form of golden yellow globules. The catechin globules increased in the structurally intact and highly turgid cells of osmotin expressing transgenic tea plants after stress treatment with 20% PEG; by contrast, the cells in non-transgenic plants accumulated fewer catechin globules. Spectrophotometric quantification of catechins also confirmed higher levels in transgenics compared to untransformed plants. We found elevated accumulation of catechins in stress tolerant cells of tea leaves.

  6. Migrating mast cells in the gallbladder epithelium of cattle and sheep. A comparative morphologic and histochemical study.

    Science.gov (United States)

    Toledo, O M; Morales, C R; Pereyra, L A; Jordão, T; Montes, G S

    1981-01-01

    This paper reports the existence of mast cells in an epithelial location in the gallbladders of both cattle and sheep. The histochemical studies performed on these cells showed that their cytoplasmic granules contain heparin and biogenic amines in both species. Optical- and electron microscopic observations demonstrated that, in both species, mast cells from the connective tissue of the gallbladder diapedese across the basal lamina and migrate through the epithelium all the way to the luminal surface, and that a degranulation process takes place during this migration. The biochemical results showed a correlation between the number of mast cells present in the epithelium and the amount of heparin detected in the different regions of the gallbladders of the species studied. Unusually high contents of heparin were found in both cattle and sheep gallbladders, suggesting that they should be studied as possible commercial sources of this polimer.

  7. Ultrastructural, histochemical, and biochemical studies of two cases with amylase, ACTH, and beta-MSH producing tumor.

    Science.gov (United States)

    Gomi, K; Kameya, T; Tsumuraya, M; Shimosato, Y; Zeze, F; Abe, K; Yoneyama, T

    1976-10-01

    Two cases of intrathoracic tumor, different in histology and accompanied by hyperamylasemia, were studied ultrastructurally, histochemically, and biochemically. The ultrastructure of the tumor cell cytoplasm showed many zymogen granules in case 1 and smaller cored granules in addition to zymogen granules in case 2. Both tumors contained not only a large amount of amylase, which was electrophoretically of saliva type with three components, but also significant amounts of immunoreactive ACTH and beta-MSH. Starch film and immunofluorescence showed that the tumor cells stored amylase. It was concluded from these findings that the tumor cells ectopically producing amylase, which showed differentiation toward the cells with zymogen production, could differentiate toward the cells of ACTH-MSH system at the same time.

  8. Enzymes, Dentinogenesis and Dental Caries: a Literature Review

    Directory of Open Access Journals (Sweden)

    Markku Larmas

    2014-12-01

    Full Text Available Objectives: Search in PubMed with keywords “enzymes, dentinogenesis, and dental caries” revealed only 4 items, but when combined with “enzymes, osteogenesis, and osteoporosis” as high as 404 items resulted. Dental caries was associated with an order of magnitude fewer studies than the chronic bone disease, osteoporosis. This observation motivated this review. Material and Methods: A comprehensive review of the available literature on role of enzymes in dentinogenesis and dental caries was undertaken using MEDLINE (PubMed and Scopus. Keywords for the search were: enzymes and odontoblasts, enzymes and different forms of dentinogenesis as well as dental caries. Results: Search revealed studies which described odontoblasts harbouring numerous enzymes (hydrolases, including metalloproteinases, transaminases and dehydrogenases during primary dentinogenesis. Alkaline phosphatase activity sharply decreased when odontoblasts turned into quiescent odontoblasts. Tertiary dentinogenesis was characterized first by reactionary dentine formation when alkaline phosphatase was highly reactivated. Then later some of these odontoblasts may die out and be replaced by other progenitor cells of pulpal origin. This tertiary dentine was called reparative dentine. Pulpal progenitor/stem cells revealed alkaline phosphatase activity in areas encircling inflamed pulp sections. Soft carious dentine revealed high hydrolase, transaminase and dehyrogenase activities that may have originated from invading microbes, saliva or were endogenous. Proteolytic activity was especially demonstrable using histochemical and biochemical means. Specifically, matrix metalloproteases may have originated partly from activated proenzymes of host origin. Conclusions: Though dental studies are scanty when compared to bone, the active role of large spectrum of enzymes in healthy and carious dentine was given support.

  9. The histochemical profile of the rat extensor digitorum longus muscle differentiates after birth and dedifferentiates in senescence

    Directory of Open Access Journals (Sweden)

    M Lehnert

    2009-08-01

    Full Text Available Age dependent motor unit dedifferentiation is a key component of impaired muscle function in advanced age. Here, we tested the hypothesis that rat muscle histochemical profile during the lifespan of an individual has an age-specific pattern since comprehensive longitudinal studies of muscle differentiation after birth and dedifferentiation in advanced age are scarce. Our results show that extensor digitorum longus muscle (EDL is comprised only of two fiber types after birth, type slow-oxidative (SO and type SDH-intermediate (SDHINT, the latter being indicative for the presence of polyneuronal innervation. In contrast to the constantly growing crosssectional area of the muscle fibers, a dramatic decrease in SDH-INT proportion occurs between day 14 and 21 after birth resulting in a complete loss of fiber type SDH-INT at the age of 90 days (p<0.05. At the age of 270 days, the fiber type composition of rat EDL dedifferentiates as shown by the reappearance of the SDH-INT type with a further increase at the age of 540 days (p<0.05. These changes in histochemical fiber type spectra are brought about by fiber type conversion within the fast twich fibers. The findings of the present study provide further evidence that fiber type conversion is a basic mechanism leading to motor unit differentiation and dedifferentiation during ontogenesis. Fiber type conversion shows a distinct time specific pattern and is also characteristic for motor unit regeneration after peripheral nerve repair. Factors that influence fiber type conversion and thereby motor unit organization may provide a future therapeutic option to enhance the regenerative capacity of motor units.

  10. Small intestinal mucosal injury in the experimental blind loop syndrome. Light- and electron-microscopic and histochemical studies.

    Science.gov (United States)

    Toskes, P P; Giannella, R A; Jervis, H R; Rout, W R; Takeuchi, A

    1975-05-01

    Microscopic (light and electron) and histochemical abnormalities have been demonstrated in the jejunum of rats with the blind loop syndrome. Three groups of animals were studied: normal control animals, and animals with either self-filling (SF) or self-emptying (SE) blind loops. Vitamin B12 malabsorption and bacterial overgrowth occurred only in those animals with SF blind loops. Three jejunal segments were studied: the blind loop segment and the jejunal segments proximal and distal to the blind loop. In the animals with the blind loop syndrome, those with SF blind loops, the most striking findings occurred in the blind loop itself, with similar but less marked changes in the jejunum distal but not proximal to the blind loop segment. Hypertrophy of both crypts and villi was evident with focal abnormalities of villus architecture. Approximately 10 to 20% of the columnar cells in the upper half of the villi were swollen and vesiculated. By electron microscopy microvilli demonstrated a variety of degeneration changes and the glycocalyx and terminal web were disrupted. Mitochondria and endoplasmic reticulum (ER), both smooth and rough, were swollen. Concentric whorls of parallel membranes and long, curvilinear rough ER were present in the cytoplasm. Histochemically, there was loss of enzymatic activity in the epithelial brush border, mitochondria and ER. Inasmuch as bacterial invasion of the jejunal mucosa was not seen, the etiology of these changes is not known but may involve bacterial "toxins" or products of bacterial metabolism. These morphological observations demonstrate that both brush border and intracellular injury occur in the jejunal epithelial cell of rats with the experimental blind loop syndrome.

  11. 氨基糖苷类修饰酶引起的细菌耐药性机制的研究进展%Deciphering Mechanisms of Aminoglycoside Antibiotics with Enzymes That Cause Resistance to Their Action

    Institute of Scientific and Technical Information of China (English)

    武灵芝; 胡栋; 秦猛

    2013-01-01

    氨基糖苷类抗生素是高效、广谱的杀菌药物.随着在临床的广泛应用,抗生素的抗药性日趋严重,这在很大程度上降低了其临床应用的潜力.其中,最主要的原因就是细菌产生了一系列修饰酶修饰抗生素的特定基团,使其失去药效.细菌产生的修饰酶种类众多,主要包括磷酸化、乙酰化和腺苷化修饰酶.研究发现,一种酶可以修饰多种抗生素,同时,一种抗生素也可以被多种修饰酶修饰.由于修饰酶底物的广谱性,使得细菌的耐药性难以克服.因此,本文就氨基糖苷类修饰酶和抗生素相互作用的热力学和动力学性质进行了详细的论述,试图找出不同修饰酶失活抗生素药物的共同作用机制.这将为设计新的抗生素药物及修饰酶抑制剂、克服细菌的耐药性,提供理论指导和技术支持.%Aminoglycosides are valuable and broad spectrum of bactericidal antibiotics. However, their therapeutic effectiveness has been severely reduced in recent decades due to the emergence of bacterial strains that are insensitive to aminoglycoside action. The most common mode of bacterial resistance to aminoglycoside antibiotics is the enzyme-catalysed chemical modification on the special groups of the drug. Aminoglycoside-modifying enzymes are widely distributed among bacterial pathogens and include O-phosphoryltransferases (kinases), N-acetyltransferases, and O-adenyltransferases. These enzymes can use several aminoglycosides as substrates regardless of size and structural differences among them. Conversely, each aminoglycoside can be a substrate for many different AGMEs. In this review, the authors describe the thermodynamic characterization of aminoglycoside modified enzyme interacted with antibiotics in an effort to define shared aspects of enzyme-aminoglycoside complexes, which provides the necessary tools and understanding to design new drugs to combat antibiotic resistance.

  12. Light microscopical localization of enzymes by means of cerium-based methods. I.V. Optimization procedures for acid phosphatase.

    Science.gov (United States)

    Halbhuber, K J; Zimmermann, N; Feuerstein, H

    1986-01-01

    The earlier described cerium based histochemical reaction for acid phosphatase [Ce-Pb-reaction, Zimmermann and Halbhuber (1985)] was optimized. The target tissues (kidney, intestine) were fixed by perfusion with glutaraldehyde in cacodylate or piperazine buffer in anesthetized animals. Postfixation of prefixed sections is not advantageous because of the detectable repressing of the enzyme activity. Moreover, the employment of unfixed cryostat sections, which were postfixed, was always connected with a complete abolition of the acid phosphatase activity. The optimal concentration of the primary capture cerium III chloride in the incubation medium is about 1 mmol. Lower concentrations lead to an incomplete histochemical detection of phosphatase activity in lysosomes. The treatment of cryostat sections of perfusion fixed tissue with borohydride (diminution of aldehyde induced cross links) or with dimethylsulfoxide (extraction of lysosomal materials or the well known vehicle property) brought about an improvement of the penetration capacity for cerium-III-cations into the target structures. After conversion of the cerium phosphate (primary specific reaction product) into cerium perhydroxide, oxalate or fluoride, the Ce-Pb-reaction was negative. Therefore, these blocking reactions represent specific inhibition controls, which indicates the formation and presence of cerium phosphate. On the basis of these reactions it is possible to check the specificity of the histochemical Ce-Pb-reaction for phosphatase activity in sections.

  13. Influence of long-term hyper-gravity on the reactivity of succinic acid dehydrogenase and NADPH-diaphorase in the central nervous system of fish: a histochemical study

    Science.gov (United States)

    Anken, R. H.; Rahmann, H.

    In the course of a densitometric evaluation, the histochemically demonstrated reactivity of succinic acid dehydrogenase (SDH) and of NADPH-diaphorase (NADPHD) was determined in different brain nuclei of two teleost fish (cichlid fish Oreochromis mossambicus, swordtail fish Xiphophorus helleri), which had been kept under 3g hyper-gravity for 8 days. SDH was chosen since it is a rate limiting enzyme of the Krebs cycle and therefore it is regarded as a marker for metabolic and neuronal activity. NADPHD reactivity reflects the activity of nitric oxide synthase. Nitric oxide (NO) is a gaseous intercellular messenger that has been suggested to play a major role in several different in vivo models of neuronal plasticity including learning. Within particular vestibulum-connected brain centers, significant effects of hyper-gravity were obtained, e.g., in the magnocellular nucleus, a primary vestibular relay ganglion of the brain stem octavolateralis area, in the superior rectus subdivision of the oculomotoric nucleus and within cerebellar eurydendroid cells, which in teleosts possibly resemble the deep cerebellar nucleus of higher vertebrates. Non-vestibulum related nuclei did not respond to hypergravity in a significant way. The effect of hyper-gravity found was much less distinct in adult animals as compared to the circumstances seen in larval fish (Anken et al., Adv. Space Res. 17, 1996), possibly due to a development correlated loss of neuronal plasticity.

  14. Enzyme detection by microfluidics

    DEFF Research Database (Denmark)

    2013-01-01

    Microfluidic-implemented methods of detecting an enzyme, in particular a DNA-modifying enzyme, are provided, as well as methods for detecting a cell, or a microorganism expressing said enzyme. The enzyme is detected by providing a nucleic acid substrate, which is specifically targeted...

  15. Histochemical studies on genetical control of hormonal enzyme inducibility in the mouse. I. Non-specific esterase activity and regional histology of the epididymis

    DEFF Research Database (Denmark)

    Blecher, S R; Kirkeby, S

    1978-01-01

    As a base line for future cell genetical studies the authors record the distribution of non-specific esterase reaction in the various histologically distinguishable cell types of the mouse epididymis. The findings are correlated with previous descriptions of the lobar structure of the organ...

  16. Ethylene response factor BnERF2-like (ERF2.4) from Brassica napus L. enhances submergence tolerance and alleviates oxidative damage caused by submergence in Arabidopsis thaliana

    Institute of Scientific and Technical Information of China (English)

    Yanyan Lv; Sanxiong Fu; Song Chen; Wei Zhang; Cunkou Qi

    2016-01-01

    Ethylene response factor proteins play an important role in regulating a variety of stress responses in plants, but their exact functions in submergence stress are not well understood. In this study, we isolated BnERF2.4 from Brassica napus L. to study its function in submergence tolerance. The expression of the BnERF2.4 gene in B. napus and the expression of antioxidant enzyme genes in transgenic Arabidopsis were analyzed by quantitative RT-PCR. The expression of BnERF2.4 was induced by submergence in B. napus and the overexpression of BnERF2.4 in Arabidopsis increased the level of tolerance to submergence and oxidative stress. A histochemical method detected lower levels of H2O2, O2•− and malondialdehyde (MDA) in transgenic Arabidopsis. Compared to the wild type, transgenic lines also had higher soluble sugar content and higher activity of antioxidant enzymes, which helped to protect plants against the oxidative damage caused by submergence. It was concluded that BnERF2.4 increased the tolerance of plants to submergence stress and may be involved in regulating soluble sugar content and the antioxidant system in defense against submergence stress.

  17. Diffusion and clearance of superparamagnetic iron oxide nanoparticles infused into the rat striatum studied by MRI and histochemical techniques

    Energy Technology Data Exchange (ETDEWEB)

    Wang, F H; Yoshitake, T; Kehr, J [Department of Physiology and Pharmacology, Karolinska Institutet, 171 77 Stockholm (Sweden); Kim, D K [Department of Materials Science, Royal Institute of Technology, 100 44 Stockholm (Sweden); Johansson, S M; Muhammed, M [Department of Neuroscience, Karolinska Institutet, 171 77 Stockholm (Sweden); Bjelke, B, E-mail: Jan.Kehr@ki.se [Department of Clinical Neuroscience, Karolinska Institutet, 171 77 Stockholm (Sweden)

    2011-01-07

    The purpose of the present study was to investigate, by MRI and histochemical techniques, the diffusion and clearance abilities of superparamagnetic iron oxide nanoparticles (SPION) coated with dextran (Dextran-SPION) and gold (Au-SPION) following their local infusions into the rat brain. In separate groups of anesthetized rats, the Dextran-SPION and Au-SPION were infused at concentrations of 0.01, 0.1, 1 and 5 {mu}g Fe/0.5 {mu}l and at the flow rate of 0.5 {mu}l min{sup -1} into the left and right striata, respectively. Repetitive T2-weighted spin-echo MRI scans were performed at time intervals of 1, 6, 12, 24, 48, 72 h, and one, two and eight weeks after inoculation. Following infusion of Dextran-SPION (0.1 {mu}g and 1 {mu}g Fe), the maximal distribution volume was observed at about 12-24 h after inoculation and two weeks later the Fe signals were undetectable for the lower dose. On the other hand, Au-SPION remained tightly localized in the closest vicinity of the infusion site as revealed by unchanged MRI signal intensities and strong histochemical staining of Fe{sup 2+} and Fe{sup 3+} ions in the corresponding brain slices. Immunohistochemical staining of astrocytic and microglial reactions revealed that there were no marked differences in GFAP, VIM or OX-42 labeling observed between the nanoparticle types, however the astrocytic reaction was more pronounced in rats receiving nanoparticles compared to the control (aCSF-infused) rats. In conclusion, the present data demonstrate that the viral-sized Dextran-SPION were able to diffuse freely through the interstitial space of the brain being progressively cleared out from the infusion site within two weeks. Thus, Dextran-SPION could be beneficially used in MRI-guided diagnostic applications such as in experimental oncology or as labels and carriers for targeted drug delivery, whereas Au-SPION could be used for labeling and tracking the transplanted stem cells in experimental MRI.

  18. AMP deaminase histochemical activity and immunofluorescent isozyme localization in rat skeletal muscle

    Science.gov (United States)

    Thompson, J. L.; Sabina, R. L.; Ogasawara, N.; Riley, D. A.

    1992-01-01

    The cellular distribution of AMP deaminase (AMPda) isozymes was documented for rat soleus and plantaris muscles, utilizing immunofluorescence microscopy and immunoprecipitation methods. AMPda is a ubiquitous enzyme existing as three distinct isozymes, A, B and C, which were initially purified from skeletal muscle, liver (and kidney), and heart, respectively. AMPda-A is primarily concentrated subsarcolemmally and intermyofibrillarly within muscle cells, while isozymes B and C are concentrated within non-myofiber elements of muscle tissue. AMPda-B is principally associated with connective tissues surrounding neural elements and the muscle spindle capsule, and AMPda-C is predominantly associated with circulatory elements, such as arterial and venous walls, capillary endothelium, and red blood cells. These specific localizations, combined with documented differences in kinetic properties, suggest multiple functional roles for the AMPda isozymes or temporal segregation of similar AMPda functions. Linkage of the AMPda substrate with adenosine production pathways at the AMP level and the localization of isozyme-C in vascular tissue suggest a regulatory role in the microcirculation.

  19. Nemaline myopathy: clinical, histochemical and immunohistochemical features Miopatia nemalínica: achados clínicos, histoquímicos e imuno-histoquímicos

    OpenAIRE

    Nazah Cherif Mohamad Youssef; Rosana Herminia Scola; Paulo José Lorenzoni; Lineu César Werneck

    2009-01-01

    Nemaline myopathy (NM) is a congenital disease that leads to hypotonia and feeding difficulties in neonates. Some cases have a more benign course, with skeletal abnormalities later in life. We analyzed a series of eight patients with NM obtained from a retrospective analysis of 4300 muscle biopsies. Patients were classified as having the typical form in five cases, intermediate form in two cases and severe form in one case. Histochemical analysis showed mixed rods distribution in all cases an...

  20. Oxidoreductases in early gestational monkey placenta during maternal malarial infection : histochemical localisation

    Directory of Open Access Journals (Sweden)

    Nishi Saxena , P.S.R. Murthy

    2007-06-01

    Full Text Available Background & objectives: Early gestational malaria is more deleterious than late gestational infection.Still the pathophysiology of maternofoetal organ—the placenta in malaria remains almost unexploredduring early gestation. Present study dealing with oxidoreductases in early gestational placenta duringmaternal malarial infection of Plasmodium cynomolgi bastianellii in rhesus monkeys was anticipatedto provide a better insight into the functional impairment of this organ leading to foetal abnormalities.Methods: Three control and four experimental monkeys (Macaca mulatta were quarantined for onemonth prior to experimentation. Experimental monkeys at 2–2½ months of gestation were inoculatedwith P. cynomolgi bastianellii. On attaining first peak of parasitaemia the placentae were collectedfrom anesthetised animals. The snap-frozen, cryostat sections were subjected to histochemicallocalisation for 3 (or 17 β-hydroxysteroid dehydrogenase (β-HSD [3 (or 17 β-hydroxysteroid:NAD (P+ oxidoreductase, EC 1.1.1.51 hydroxysteroid dehydrogenases] and NADPH-tetrazoliumreductase [NADPH : (acceptor oxidoreductase, EC 1.6.99.1 NADPH-TR]. Comparative microscopyof control and malaria infected placental sections was performed and analysed.Results: A localised decrease in both the enzymes was observed in syncytiotrophoblast layer ofmalaria infected monkey placenta. The areas showing morphological damage of syncytiotrophoblastwere also depicting gross reduction in NADPH-TR activity.Interpretation & conclusion: The altered enzymatic activities [3 (or 17 β-HSD and NADPH-TR] inmalaria infected early gestational monkey placenta have been discussed in the light of placentalfunction. It could be concluded by present studies that these alterations would affect the cellularmetabolism especially steroidogenesis and detoxification process which in turn would affect thenormal development of the foetus as well as maintenance of gestation.

  1. Histological and histochemical study of the protective role of rosemary extract against harmful effect of cell phone electromagnetic radiation on the parotid glands.

    Science.gov (United States)

    Ghoneim, Fatma M; Arafat, Eetmad A

    2016-06-01

    Electromagnetic fields (EMFs) are a class of non-ionizing radiation (NIR) that is emitted from mobile phone. It may have hazardous effects on parotid glands. So, we aimed to investigate the histological and histochemical changes of the parotid glands of rats exposed to mobile phone and study the possible protective role of rosemary against its harmful effect. Forty adult male albino rats were used in this study. They were classified into 4 equal groups. Group I (control), group II (control receiving rosemary), group III (mobile phone exposed group) and group IV (mobile exposed, rosemary treated group). Parotid glands were dissected out for histological and histochemical study. Moreover, measurement of oxidative stress markers; malondialdehyde (MDA) and total antioxidant capacity (TAC) was done. The results of this study revealed that rosemary has protective effect through improving the histological and histochemical picture of the parotid gland in addition of its antioxidant effect. It could be concluded from the current study, that exposure of parotid gland of rat models to electromagnetic radiation of mobile phone resulted in structural changes at the level of light and electron microscopic examination which could be explained by oxidative stress effect of mobile phone. Rosemary could play a protective role against this harmful effect through its antioxidant activity.

  2. Characteristics of alveolar bone associated with physiological movement of molar in mice: a histological and histochemical study.

    Science.gov (United States)

    Matsuda, Kie; Haga-Tsujimura, Maiko; Yoshie, Sumio; Shimomura-Kuroki, Junko

    2014-01-01

    Mouse molars undergo distal movement, during which new bone is formed at the mesial side of the tooth root whereas the preexisting bone is resorbed at the distal side of the root. However, there is little detailed information available regarding which of the bones that surround the tooth root are involved in physiological tooth movement. In the present study, we therefore aimed to investigate the precise morphological differences of the alveolar bone between the bone formation side of the tooth root, using routine histological procedures including silver impregnation, as well as by immunohistochemical analysis of alkaline phosphatase and tartrate-resistant acid phosphatase activity, and immunohistochemical analysis of the expression of the osteocyte markers dentin matrix protein 1, sclerostin, and fibroblast growth factor 23. Histochemical analysis indicated that bone formation by osteoblasts and bone resorption by osteoclasts occurred at the bone formation side and the bone resorption side, respectively. Osteocyte marker immunoreactivity of osteocytes at the surface of the bone close to the periodontal ligament differed at the bone formation and bone resorption sides. We also showed different specific features of osteocytic lacunar canalicular systems at the bone formation and bone resorption sides by using silver staining. This study suggests that the alveolar bone is different in the osteocyte nature between the bone formation side and the bone resorption side due to physiological distal movement of the mouse molar.

  3. Histochemical screening, metabolite profiling and expression analysis reveal Rosaceae roots as the site of flavan-3-ol biosynthesis.

    Science.gov (United States)

    Hoffmann, T; Friedlhuber, R; Steinhauser, C; Tittel, I; Skowranek, K; Schwab, W; Fischer, T C

    2012-01-01

    Histochemical screening of 30 Rosaceae genera representing all classic subfamilies demonstrated flavan-3-ols (catechins) as general secondary metabolites in roots of Rosaceae. Semi-quantitative LC-MS analyses confirmed the presence of catechin, epicatechin and various dimeric flavan-3-ols (also representing higher polymeric proanthocyanidins) as prominent polyphenols in root tips of Fragaria (strawberry), Malus (apple), Rosa (rose), Pyrus (pear) and Prunus (plum). Distinct patterns of flavan-3-ol distribution at the cellular level were found in strawberry (Fragaria × ananassa) and apple (Malus × domestica) root tips. The calyptras (root caps) showed the most prominent flavan-3-ol staining for these two genera. Border cells of Fragaria and Malus, as first demonstrated here for Rosaceae, were also found to contain flavan-3-ols. Transcript analyses with cDNA demonstrated root expression of known flavonoid genes expressed in the respective fruits and leaves. Primarily, this proves in situ biosynthesis of flavan-3-ols in these roots. Knowledge of the distinct cellular distribution patterns and their in situ biosynthesis in roots provides a basis for analysis of the functional roles of Rosaceae root flavan-3-ols.

  4. Classification of 27 Tumor-Associated Antigens by Histochemical Analysis of 36 Freshly Resected Lung Cancer Tissues.

    Science.gov (United States)

    Kurosawa, Gene; Sugiura, Mototaka; Hattori, Yoshinobu; Tsuda, Hiroyuki; Kurosawa, Yoshikazu

    2016-11-08

    In previous studies, we identified 29 tumor-associated antigens (TAAs) and isolated 488 human monoclonal antibodies (mAbs) that specifically bind to one of the 29 TAAs. In the present study, we performed histochemical analysis of 36 freshly resected lung cancer tissues by using 60 mAbs against 27 TAAs. Comparison of the staining patterns of tumor cells, bronchial epithelial cells, and normal pulmonary alveolus cells and interalveolar septum allowed us to determine the type and location of cells that express target molecules, as well as the degree of expression. The patterns were classified into 7 categories. While multiple Abs were used against certain TAAs, the differences observed among them should be derived from differences in the binding activity and/or the epitope. Thus, such data indicate the versatility of respective clones as anti-cancer drugs. Although the information obtained was limited to the lung and bronchial tube, bronchial epithelial cells represent normal growing cells, and therefore, the data are informative. The results indicate that 9 of the 27 TAAs are suitable targets for therapeutic Abs. These 9 Ags include EGFR, HER2, TfR, and integrin α6β4. Based on our findings, a pharmaceutical company has started to develop anti-cancer drugs by using Abs to TfR and integrin α6β4. HGFR, PTP-LAR, CD147, CDCP1, and integrin αvβ3 are also appropriate targets for therapeutic purposes.

  5. Classification of 27 Tumor-Associated Antigens by Histochemical Analysis of 36 Freshly Resected Lung Cancer Tissues

    Directory of Open Access Journals (Sweden)

    Gene Kurosawa

    2016-11-01

    Full Text Available In previous studies, we identified 29 tumor-associated antigens (TAAs and isolated 488 human monoclonal antibodies (mAbs that specifically bind to one of the 29 TAAs. In the present study, we performed histochemical analysis of 36 freshly resected lung cancer tissues by using 60 mAbs against 27 TAAs. Comparison of the staining patterns of tumor cells, bronchial epithelial cells, and normal pulmonary alveolus cells and interalveolar septum allowed us to determine the type and location of cells that express target molecules, as well as the degree of expression. The patterns were classified into 7 categories. While multiple Abs were used against certain TAAs, the differences observed among them should be derived from differences in the binding activity and/or the epitope. Thus, such data indicate the versatility of respective clones as anti-cancer drugs. Although the information obtained was limited to the lung and bronchial tube, bronchial epithelial cells represent normal growing cells, and therefore, the data are informative. The results indicate that 9 of the 27 TAAs are suitable targets for therapeutic Abs. These 9 Ags include EGFR, HER2, TfR, and integrin α6β4. Based on our findings, a pharmaceutical company has started to develop anti-cancer drugs by using Abs to TfR and integrin α6β4. HGFR, PTP-LAR, CD147, CDCP1, and integrin αvβ3 are also appropriate targets for therapeutic purposes.

  6. Ovary organization and oogenesis in the tardigrade Macrobiotus polonicus Pilato, Kaczmarek, Michalczyk & Lisi, 2003 (Eutardigrada, Macrobiotidae): ultrastructural and histochemical analysis.

    Science.gov (United States)

    Poprawa, Izabela; Schlechte-Wełnicz, Weronika; Hyra, Marta

    2015-05-01

    The female reproductive system, the process of oogenesis, and the morphology of the egg capsule of Macrobiotus polonicus were analyzed using transmission and scanning electron microscopy and histochemical methods. The female reproductive system of Macrobiotus polonicus consists of a single ovary and a single oviduct that opens into the cloaca. The seminal receptacle filled with sperm cells is present. The ovary is divided into two parts: a germarium that is filled with oogonia and a vitellarium that is filled with branched clusters of the germ cells. Meroistic oogenesis occurs in the species that was examined. The yolk material is synthesized by the oocyte (autosynthesis) and by the trophocytes and is transported to the oocyte through cytoplasmic bridges. The process of the formation of the egg envelopes starts in the late vitellogenesis. The egg capsule is composed of two envelopes-the vitelline envelope and the three-layered chorion. The vitelline envelope is of the primary type while the chorion is of a secondary type. The surface of the chorion is covered with conical processes that terminate with a strongly indented terminal disc.

  7. Lectin-histochemical reactivity of sialic acid in breast cancer and its relationship to prognosis using limulus polyphemus agglutinin.

    Science.gov (United States)

    Ding, K; Yamaguchi, A; Goi, T; Maehara, M; Nakagawara, G

    1997-04-01

    Studies of circulating sialic acid have revealed its relationship with a variety of malignant tumors. It is not vet clear whether sialic acid could be used as a prognostic marker of breast cancer, and few studies have examined sialic acid expression in the cell membrane and cytoplasm of breast cancer cells by means of the lectin-histochemical technique. In the present study, we used biotinylated limulus polyphemus agglutinin (LPA), a special binding lectin of sialic acid, to stain sialic acid in breast cancer cells. Of the 104 cases of breast cancer examined, 59 (56.7%) positive cases were observed. There was a significant correlation between the LPA staining and the clinicopathologic features of all patients, including pathological stage and lymph node metastasis. Among the 100 patients who underwent curative operation, the mean disease-free survival rate of the 45 patients who were LPA-negative was significantly higher than that of the 55 LPA-positive patients (psialic acid in breast cancer could be used as a marker of malignancy potential, as well as a poor survival factor, and the biotinylated LPA assay may provide a convenient and useful method to predict the prognosis of breast cancer.

  8. Morphological characteristics of the digestive tract of Schizodon knerii (Steindachner, 1875, (Characiformes: Anostomidae: An anatomical, histological and histochemical study

    Directory of Open Access Journals (Sweden)

    Marcella L. Dos Santos

    2015-06-01

    Full Text Available The digestive tracts of 44 specimens of Schizodon knerii were studied using anatomical, histological and histochemical techniques. The mouth has terminal position, the lip epithelium is squamous stratified with mucous, claviform cells and taste buds, teeth have an incisive form and the tongue has a stratified squamous epithelium with mucous cells and taste buds. The oropharynx cavity is formed by gill apparatus and pharyngeal teeth. The oesophagus presented pleated mucosa, a stratified squamous epithelium with mucous cells, oesophageal glands and taste buds. The stomach presented cardiac, fundic and pyloric regions, simple prismatic epithelium with tubular glands, with none in the pyloric region. The intestine contains 11-15 pyloric caeca, a simple prismatic epithelium with brush border, goblet cells and lymphocytes. Mucosal cells, oesophageal glands and goblet cells reacted positively to PAS, amylase + PAS, Ab pH 2.5 and Ab pH 0.5. Gastric prismatic cells reacted positively to PAS, amylase + PAS, but only those in the pyloric region reacted positively to Ab pH 2.5 and Ab pH 0.5. The results improve the understanding of the anatomy of S. knerii feeding habits and the presence of mucosubstances in the epithelium, highlights the importance of glycoproteins for passing food through the digestive tract.

  9. Mucinous carcinoma of the breast. A clinicopathologic, histochemical, and immunocytochemical study with special reference to neuroendocrine differentiation.

    Science.gov (United States)

    Scopsi, L; Andreola, S; Pilotti, S; Bufalino, R; Baldini, M T; Testori, A; Rilke, F

    1994-07-01

    We studied the clinical, histologic, histochemical, and immunocytochemical characteristics of 61 mucinous tumors (38 pure, 23 mixed) retrieved from a consecutive series of 1,689 infiltrating carcinomas of the female breast. The only statistically significant predictors of favorable survival were histologic (pure) type coupled with the absence of axillary lymph node metastases. Other factors, including classification into A and B types according to Capella et al., and neuroendocrine status, as assessed by the presence of argyrophilia, granins, neuron-specific enolase (NSE), and synaptophysin (SYN),-all had no influence on survival. Argyrophilic cells were found in 16 pure mucinous tumors (42%) and in the mucinous component of four mixed tumors (17%). Granin (chromogranin A or B), NSE, and SYN immunoreactivities were demonstrated in all the argyrophilic tumors. We also found NSE- and SYN-immunoreactive cells in 31 of 41 and 16 of 41 nonargyrophilic (granin-unreactive) mucinous tumors, which supports the view that mucinous carcinomas of the breast as a whole are neuroendocrine-programmed tumors.

  10. Regional potassium distribution in the brain in forensic relevant types of intoxication preliminary morphometric evaluation using a histochemical method.

    Science.gov (United States)

    Oehmichen, M; Ochs, U; Meissner, C

    2001-02-01

    A histochemical-morphometric method was used to measure potassium (K+) levels in gray and white matter of rats following sublethal intoxication with 11 different neurotoxic compounds of high forensic significance. Six rats were each given a single substance applied intraperitoneally, the same dosage being given to two animals each. The animals were subsequently killed, the brains immediately frozen, and cryosections cut. K+ levels were evaluated morphometrically. A drop in K+ levels was used as the criterion for cytotoxic edema. Application of ethanol, atropine, carbromal, carbon monoxide, morphine or triethyltin led to a rise in K+ levels in the gray matter and a simultaneous decline in the white matter. By contrast, administration of amitriptyline, glycerin, potassium cyanide, parathion or phenobarbital initiated an increase in K+ levels in both gray and white matter. A cytotoxic edema could thus be reliably excluded in these intoxications. Although the study design allows no statistical analysis, these conclusions are supported by the marked differences in K+ levels in gray and white matter induced by the different toxicants.

  11. Classification of 27 Tumor-Associated Antigens by Histochemical Analysis of 36 Freshly Resected Lung Cancer Tissues

    Science.gov (United States)

    Kurosawa, Gene; Sugiura, Mototaka; Hattori, Yoshinobu; Tsuda, Hiroyuki; Kurosawa, Yoshikazu

    2016-01-01

    In previous studies, we identified 29 tumor-associated antigens (TAAs) and isolated 488 human monoclonal antibodies (mAbs) that specifically bind to one of the 29 TAAs. In the present study, we performed histochemical analysis of 36 freshly resected lung cancer tissues by using 60 mAbs against 27 TAAs. Comparison of the staining patterns of tumor cells, bronchial epithelial cells, and normal pulmonary alveolus cells and interalveolar septum allowed us to determine the type and location of cells that express target molecules, as well as the degree of expression. The patterns were classified into 7 categories. While multiple Abs were used against certain TAAs, the differences observed among them should be derived from differences in the binding activity and/or the epitope. Thus, such data indicate the versatility of respective clones as anti-cancer drugs. Although the information obtained was limited to the lung and bronchial tube, bronchial epithelial cells represent normal growing cells, and therefore, the data are informative. The results indicate that 9 of the 27 TAAs are suitable targets for therapeutic Abs. These 9 Ags include EGFR, HER2, TfR, and integrin α6β4. Based on our findings, a pharmaceutical company has started to develop anti-cancer drugs by using Abs to TfR and integrin α6β4. HGFR, PTP-LAR, CD147, CDCP1, and integrin αvβ3 are also appropriate targets for therapeutic purposes. PMID:27834817

  12. Enzyme inhibition by iminosugars

    DEFF Research Database (Denmark)

    López, Óscar; Qing, Feng-Ling; Pedersen, Christian Marcus

    2013-01-01

    Imino- and azasugar glycosidase inhibitors display pH dependant inhibition reflecting that both the inhibitor and the enzyme active site have groups that change protonation state with pH. With the enzyme having two acidic groups and the inhibitor one basic group, enzyme-inhibitor complexes...

  13. A patient with typical clinical features of mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS but without an obvious genetic cause: a case report

    Directory of Open Access Journals (Sweden)

    Abu-Amero Khaled K

    2009-10-01

    Full Text Available Abstract Introduction There are currently 23 missense point mutations and one 4 basepair deletion spanning different mitochondrial genes associated with mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS. The spectrum of mitochondrial DNA mutations in Arab patients with MELAS is largely unknown. Case presentation A standard clinical examination was carried out on a 34-year-old Saudi woman showing clinical features of MELAS. Fresh frozen muscle tissue was subjected to enzyme histochemical analysis. DNA was extracted from her leukocytes and muscle tissue, and the full mitochondrial genome was screened for base substitution mutations and deletions. Additionally, we screened the polymerase gamma-1 nuclear gene for mutations. The patient was negative for the most common m.3243 A>G MELAS mutation. Sequencing the full mitochondrial genome did not reveal any known or potentially pathogenic sequence changes. The polymerase gamma-1 gene was also free from mutations. Conclusion The clinical picture described here typically fits that observed in patients with MELAS or mitochondrial stroke-like events, but mutations in recognized genes (mitochondrial DNA and polymerase gamma-1 gene were absent. We report the case of a patient with typical clinical features of MELAS, but without an obvious genetic cause.

  14. Phenotypic and histochemical traits of the interaction between Plasmopara viticola and resistant or susceptible grapevine varieties

    Directory of Open Access Journals (Sweden)

    Toffolatti Silvia

    2012-08-01

    Full Text Available Abstract Background Grapevine downy mildew, caused by Plasmopara viticola, is a very serious disease affecting mainly Vitis vinifera cultivated varieties around the world. Breeding for resistance through the crossing with less susceptible species is one of the possible means to reduce the disease incidence and the application of fungicides. The hybrid Bianca and some of its siblings are considered very promising but their resistance level can vary depending on the pathogen strain. Moreover, virulent strains characterized by high fitness can represent a potential threat to the hybrid cultivation. Results The host response and the pathogen virulence were quantitatively assessed by artificially inoculating cv Chardonnay, cv Bianca and their siblings with P. viticola isolates derived from single germinating oospores collected in various Italian viticultural areas. The host phenotypes were classified as susceptible, intermediate and resistant, according to the Area Under the Disease Progress Curve caused by the inoculated strain. Host responses in cv Bianca and its siblings significantly varied depending on the P. viticola isolates, which in turn differed in their virulence levels. The fitness of the most virulent strain did not significantly vary on the different hybrids including Bianca in comparison with the susceptible cv Chardonnay, suggesting that no costs are associated with virulence. Among the individual fitness components, only sporangia production was significantly reduced in cv Bianca and in some hybrids. Comparative histological analysis revealed differences between susceptible and resistant plants in the pathogen diffusion and cytology from 48 h after inoculation onwards. Defence mechanisms included callose depositions in the infected stomata, increase in peroxidase activity, synthesis of phenolic compounds and flavonoids and the necrosis of stomata and cells immediately surrounding the point of invasion and determined alterations in

  15. Early morphological and histochemical alterations in rats subjected to ileocystoplasty Alterações morfológicas e histoquímicas precoces em ratos submetidos à ileocistoplastia

    Directory of Open Access Journals (Sweden)

    Edinaldo Gonçalves de Miranda

    2009-10-01

    Full Text Available PURPOSE: To study morphologic and histochemical alterations arising at the ileocystoplasty site. METHODS: Sixteen Wistar female rats were subjected to ileocystoplasty and sacrificed after eight weeks. Material collected was divided into four groups for histological and histochemical studies: Group I (control - isolated ileum segment removed during ileocystoplasty; Group II - ileoileal anastomosis; Group III - ileovesical anastomosis and Group IV - ileal segment from the neobladder. Histological and histochemical study assessed dysplasia, metaplasia, acute and chronic inflammation, fibrosis, atrophy, hypertrophy, total mucins, sialomucins and sulfomucins. The non-parametric Wilcoxon and Mann-Whitney tests were employed in statistical analysis. RESULTS: None of the groups presented dysplasia. Acute inflammation and atrophy occurred in Groups II, III and IV, not reaching statistical significance. Metaplasia was significant only in Group III (p=0.012. Chronic inflammation, fibrosis and hypertrophy were significant in Groups II, III and IV. There was a significant increase in total mucin content in Group IV (p=0.014 and a reduction in Group III (p=0.016. Increases in sialomucins were observed in samples for Groups III (p=0.003 and IV (p=0.002 along with reduced sulfomucins in samples from Groups III (p=0.013 and IV (p=0.008. CONCLUSION: Ileocystoplasty in female rats caused squamous metaplasia, chronic inflammatory infiltration, fibrosis, hypertrophy, increase in sialomucin content, reduction in sulfomucins, and alterations in total mucin content with statistical significance, as well acute inflammatory infiltration and muscular atrophy with less intensity.OBJETIVO: Estudar alterações morfológicas e histoquímicas nas ileocistoplastias em ratos fêmea. MÉTODOS: 16 ratos fêmea foram submetidos à ileocistoplastia, sacrificadas após oito semanas. O material coletado foi e dividido em quatro grupos para análise morfológica e histoquímica: Grupo

  16. Enzyme kinetics of conjugating enzymes: PAPS sulfotransferase.

    Science.gov (United States)

    James, Margaret O

    2014-01-01

    The sulfotransferase (SULT) enzymes catalyze the formation of sulfate esters or sulfamates from substrates that contain hydroxy or amine groups, utilizing 3'-phosphoadenosyl-5'-phosphosulfate (PAPS) as the donor of the sulfonic group. The rate of product formation depends on the concentrations of PAPS and substrate as well as the sulfotransferase enzyme; thus, if PAPS is held constant while varying substrate concentration (or vice versa), the kinetic constants derived are apparent constants. When studied over a narrow range of substrate concentrations, classic Michaelis-Menten kinetics can be observed with many SULT enzymes and most substrates. Some SULT enzymes exhibit positive or negative cooperativity during conversion of substrate to product, and the kinetics fit the Hill plot. A characteristic feature of most sulfotransferase-catalyzed reactions is that, when studied over a wide range of substrate concentrations, the rate of product formation initially increases as substrate concentration increases, then decreases at high substrate concentrations, i.e., they exhibit substrate inhibition or partial substrate inhibition. This chapter gives an introduction to sulfotransferases, including a historical note, the nomenclature, a description of the function of SULTs with different types of substrates, presentation of examples of enzyme kinetics with SULTs, and a discussion of what is known about mechanisms of substrate inhibition in the sulfotransferases.

  17. Tailoring and histochemical application of fluorescent homo-dimeric styryl dyes using frozen sections: from peroxidase substrates to new cytochemical probes for mast cells, keratin, cartilage and nucleic acids.

    Science.gov (United States)

    Krieg, Reimar; Eitner, Annett; Halbhuber, Karl-Jürgen

    2011-11-01

    Homo-dimers of styryl dyes were chemically tailored in order to become specific cytochemical probes for use in the life sciences. Histochemical applications using fixed cryotome sections are discussed. It is concluded, that homo-dimerization of specific styryl substrates of peroxidase (PO) by way of their covalent linkage, does not necessarily lead to improved detection sensitivity of endogenous and immuno-bound peroxidase (PO) activity. In general, these dimers act less specific towards PO activity than parent monomers. Synergetic interactions of the doubled basic dye compartments with cell constituents cause a pronounced staining of further targets at the cellular level. This behavior depends on the functional groups present in each dye compartment in a crucial manner. However, by way of chemical dye tailoring centering of these initially unwanted staining properties is possible leading to novel highly fluorescent stains for mast cells, nucleic acids, keratin and cartilage tissue. Structure/staining behavior-relationships of these stains are discussed. Copyright © 2011 Elsevier GmbH. All rights reserved.

  18. Histological and histochemical effects of Gly-phosate on testicular tissue and function

    Directory of Open Access Journals (Sweden)

    Nejati

    2012-01-01

    Full Text Available Background: In this study we aimed to evaluate the impact of chronic exposure to the Gly-phosate (GP on rat’s testicular tissue and sperm parameters.Objective: Testicular tissue, morphology of sperms and testosterone level in serum of mature male rats were analyzed.Materials and Methods: Animals were divided into two test and control-sham groups. The test group was subdivided into 4 groups (10, 20, 30 and 40 days GP administrated. Each test group (n=8 received the compound at dose of 125 mg/kg, once a day, orally for 40 days while control-sham group (n=16 received the corn oil (0.2 ml/day.Results: Microscopic analyses revealed increased thickness of tunica albuginea, obvious edema in sub-capsular and interstitial connective tissue, atrophied seminiferous tubules, arrested spermatogenesis, negative tubular differentiation and repopulation indexes, decreased Leydig cells/mm2 of interstitial tissue, hypertrophy and cytoplasmic granulation of Leydig cells, elevated death, immature sperm and increased immotile and abnormal sperm percentage. The carbohydrate ratio was reduced in first three layers of the germinal epithelium (GE cytoplasm. The upper layers of the GE series were manifested with low rate of lipid accumulation in cytoplasm, while the cells which were located in first layers were revealed with higher amount of lipid foci. Hematological investigations showed significant (p<0.05 decreasing of testosterone level in serum.Conclusion: The current data provide inclusive histological feature of chronic exposure against GP with emphasizing on reproductive disorders including histological adverse effect on the testicular tissue, spermatogenesis, sperm viability and abnormality which potentially can cause infertility

  19. What do we know about the structure of human thymic Hassall's corpuscles? A histochemical, immunohistochemical, and electron microscopic study.

    Science.gov (United States)

    Mikušová, Renáta; Mešťanová, Veronika; Polák, Štefan; Varga, Ivan

    2017-05-01

    Hassall's corpuscles are the most prominent structures in the human thymus. However, relatively few analyses have been performed to determine their function and cellular origins during development. In this study, we evaluated the cellular microenvironment of human thymic Hassall's corpuscles using histochemistry, immunohistochemistry, and transmission electron microscopy. We examined 95 human thymic tissue samples, which were perioperatively obtained from children undergoing cardiac surgery. To characterize the complex cellular microenvironment of human thymic corpuscles, we used a panel of 14 different antibodies to identify discrete cell types. We also utilized various histochemical methods (PAS reaction, alcian blue staining, alkaline phosphatase and acid phosphatase activity staining, von Kossa staining of calcified particles) and transmission electron microscopy to visualize these structures. Considerable variation in the sizes, shapes, and numbers of Hassall's corpuscles was observed, even amongst children of the same age. Inside the largest Hassall's corpuscles, cystic dilatation with an accumulation of cellular debris was found. These morphological observations might be associated with disruptions in the formation, migration, or differentiation of cardiac neural crest cells, which are essential for heart and thymus development. Immunohistochemical staining and electron microscopy revealed that Hassall's corpuscles resemble other types of stratified squamous epithelia. Most Hassall's corpuscles are heterocellular, consisting of thymic epithelial cells, macrophages, interdigitating dendritic cells, myoid cells, and, occasionally, mast cells and lymphocytes. To explore the potential functions of Hassall's corpuscles, we found that the concentrations of B-lymphocytes and BCL2-positive lymphocytes suggested a role in regulation of lymphopoiesis. We also found that these structures do not originate from the perivascular epithelium as previously proposed, nor

  20. [Histochemical properties of secretory granules and fine structure of terminal portion in the Japanese Macaque labial gland].

    Science.gov (United States)

    Tsutusumi, T; Kurabuchi, S; Aiyama, S

    1989-06-01

    The terminal portion of the Japanese macaque (Macaca fuscata) labial gland was examined ultrastructurally and histochemically. The results obtained are as follows. 1) The Japanese macaque, Macaca fuscata has the upper and the lower labial glands, which can be described as a compound tubulo-acinar gland. 2) The terminal portion of the labial gland appears to be consisted of the mucous acini and the demilunes when examined with the light microscope. 3) The secretory granules containing in the glandular cells of the mucous acini and the demilunes are negative with napthol yellow S, ninhydrin-Schiff and DMAB nitrite. 4) The secretory granules containing in the glandular cells of mucous acini stain intensely with PAS, alcian blue (pH1.0, 2.5, 3.5), colloidal iron and PA-methenamine silver, while those of demilunes are negative with alcian blue (pH1.0). The glandular cells of demilune with the PA-methenamine silver method shows weak positive granules and positive granules which are limited to the hallo of them. 5) In the mucous acini the mucous granules are ejected from glandular cells by the process of exocytosis. 6) The myoepithelial cell can be seen in the terminal portion. This cell surrounds the acini with long processes. These findings suggest that the glandular cells of the demilune have the granules containing mucopoly saccharides and a small quantity of protein in addition to the mucous granules, although the terminal portion of the Japanese macaque labial gland is nearly composed of mucous cells.

  1. Histochemical and biochemical analysis of the size-dependent nanoimmunoresponse in mouse Peyer’s patches using fluorescent organosilica particles

    Science.gov (United States)

    Awaad, Aziz; Nakamura, Michihiro; Ishimura, Kazunori

    2012-01-01

    Background/objective The size-dependent mucosal immunoresponse against nanomaterials (nanoimmunoresponse) is an important approach for mucosal vaccination. In the present work, the size-dependent nanoimmunoresponse of mouse Peyer’s patches (PPs) and immunoglobulin A (IgA) level was investigated using fluorescent thiol-organosilica particles. Methods Various sizes of fluorescent thiol-organosilica particles (100, 180, 365, 745, and 925 nm in diameter) were administered orally. PPs were analyzed histochemically, and IgA levels in PP homogenates, intestinal secretions around PPs, and bile were analyzed biochemically. Results When compared with the larger particles (745 and 925 nm), oral administration of smaller thiol-organosilica particles (100, 180, and 365 nm) increased the number of CD11b+ macrophages and IgA+ cells in the subepithelial domes of the PPs. Additionally, administration of larger particles induced the expression of alpha-L-fucose and mucosal IgA on the surface of M cells in the follicle-associated epithelia of PPs and increased the number of 33D1+ dendritic cells in the subepithelial domes of the PPs. IgA contents in the bile and PP homogenates were high after the administration of the 100 nm particles, but IgA levels in the intestinal secretions were high after the administration of the 925 nm particles. Two size-dependent routes of IgA secretions into the intestinal lumen, the enterohepatic route for smaller particles and the mucosal route for larger particles were proposed. Conclusion Thiol-organosilica particles demonstrated size-dependent nanoimmunoresponse after oral administration. The size of the particles may control the mucosal immunity in PPs and were useful in mucosal vaccination approaches. PMID:22619503

  2. Enzymes for improved biomass conversion

    Science.gov (United States)

    Brunecky, Roman; Himmel, Michael E.

    2016-02-02

    Disclosed herein are enzymes and combinations of the enzymes useful for the hydrolysis of cellulose and the conversion of biomass. Methods of degrading cellulose and biomass using enzymes and cocktails of enzymes are also disclosed.

  3. Enzymes for improved biomass conversion

    Energy Technology Data Exchange (ETDEWEB)

    Brunecky, Roman; Himmel, Michael E.

    2016-02-02

    Disclosed herein are enzymes and combinations of the enzymes useful for the hydrolysis of cellulose and the conversion of biomass. Methods of degrading cellulose and biomass using enzymes and cocktails of enzymes are also disclosed.

  4. Profiling the orphan enzymes

    Science.gov (United States)

    2014-01-01

    The emergence of Next Generation Sequencing generates an incredible amount of sequence and great potential for new enzyme discovery. Despite this huge amount of data and the profusion of bioinformatic methods for function prediction, a large part of known enzyme activities is still lacking an associated protein sequence. These particular activities are called “orphan enzymes”. The present review proposes an update of previous surveys on orphan enzymes by mining the current content of public databases. While the percentage of orphan enzyme activities has decreased from 38% to 22% in ten years, there are still more than 1,000 orphans among the 5,000 entries of the Enzyme Commission (EC) classification. Taking into account all the reactions present in metabolic databases, this proportion dramatically increases to reach nearly 50% of orphans and many of them are not associated to a known pathway. We extended our survey to “local orphan enzymes” that are activities which have no representative sequence in a given clade, but have at least one in organisms belonging to other clades. We observe an important bias in Archaea and find that in general more than 30% of the EC activities have incomplete sequence information in at least one superkingdom. To estimate if candidate proteins for local orphans could be retrieved by homology search, we applied a simple strategy based on the PRIAM software and noticed that candidates may be proposed for an important fraction of local orphan enzymes. Finally, by studying relation between protein domains and catalyzed activities, it appears that newly discovered enzymes are mostly associated with already known enzyme domains. Thus, the exploration of the promiscuity and the multifunctional aspect of known enzyme families may solve part of the orphan enzyme issue. We conclude this review with a presentation of recent initiatives in finding proteins for orphan enzymes and in extending the enzyme world by the discovery of new

  5. Unhairing with enzymes

    OpenAIRE

    Crispim, A.; Mota, M.

    2003-01-01

    The use of enzymes in the leather industry is increasing and their application is being widened to include operations such as de-greasing, unhairing and other wet-end operations. Enzymes can also be used to assist with recycling leather wastes as well as to avoid pollution. The present work is devoted to illustrate the potential application of enzymes in unhairing without hair destruction. Enzymatic unhairing is based upon the weakening of the epidermis basal layer to which the hair is at...

  6. HISTOCHEMICAL STUDY OF THE AFFECTION OF TEMPORARY CEREBRAL ISCHEMIA ON THE CEREBELLAR CORTEX%短暂性缺血对小脑皮质影响的组织化学研究

    Institute of Scientific and Technical Information of China (English)

    李仁; 顾远清; 陈显斌; 李昊; 熊希凯; 吴刚

    2001-01-01

    In order to explore the affection of temporary cerebral ischemiaon the purkinje cells of the cerebellary cortex. Using histochemical method, enzymehistochemical changes was observed in the purkinje cells of the rabbit after ischemia of 5 minutes (group B)、 10 minutes (group C) and reperfusion (group D、E). The results showed that after 10 minutes of ischemia and reperfusion. The activites of SDH、Mg2+-ATP、PAS were reduced (P<0.05), While the activites of LDH wre increased (P<0.01). The results suggested that the injury of 10 minutes ischemia and reperfusion may damage the activites of enzymes associated with energic metabolism in cerebellar cortex of purkinje cells.%为了探讨全脑短暂性缺血对小脑皮质蒲肯野细胞的影响,实验用组织化学方法对家兔全脑缺血5分钟(B组)、10分钟(C组)及缺血再灌(D、E组)后蒲肯野细胞的酶组织化学变化进行了观察。结果显示,缺血10分钟及缺血再灌后蒲肯野细胞的SDH、Mg2+-ATP活性及PAS反应均降低(P<0.05),LDH增高(P<0.01)。结果提示家兔全脑缺血10分钟和缺血再灌可损害小脑皮质蒲肯野细胞的能量代谢酶的活性。

  7. Modulating Effect of Hypnea musciformis (Red Seaweed) on Lipid Peroxidation, Antioxidants and Biotransforming Enzymes in 7,12-Dimethylbenz (a) Anthracene Induced Mammary Carcinogenesis in Experimental Animals

    Science.gov (United States)

    Balamurugan, Mohan; Sivakumar, Kathiresan; Mariadoss, Arokia Vijaya Anand; Suresh, Kathiresan

    2017-01-01

    Background: Breast cancer is the second most widespread diagnosed cancer and second leading cause of cancer death in women. Objective: The present work was carried out to evaluate the chemo preventive potential of Hypnea musciformis (ethanol extract) seaweed on oxidative stress markers, bio transforming enzymes, incidence of tumors, and pathological observation in 7,12-dimethylbenzanthracene (DMBA) exposed experimental mammary carcinogenesis. Materials and Methods: Female Sprague–Dawley rats were randomly divided into four groups. Rats in the group 1 served as control. Rats in the group 2 and 3 received a single subcutaneous injection of DMBA (25 mg/kg body weight (b.w)) in the mammary gland to develop mammary carcinoma. In addition, group 3 rats were orally administrated with 200 mg/kg between of H. musciformis along with DMBA injection and group 4 rats received ethanolic extract of H. musciformis every day orally (200 mg/kg b.w) throughout the experimental period of 16 weeks. Results: Our results revealed that treatment with H. musciformis ethanolic extract to DMBA treated rats significantly reduced the incidence of tumor and tumor volume as compared to DMBA alone treated rats. Moreover, our results showed imbalance in the activities/levels of lipid peroxidation by products, antioxidant enzymes, and bio transforming phase I and II enzymes in the circulation, liver and mammary tissues of DMBA treated rats which were significantly modulated to near normal on treatment with ethanolic extract of H. musciformis. All these alterations were supported by histochemical findings. Conclusion: The results obtained from this study suggest that chemo preventive potential of H. musciformis ethanol extract is probably due to their free radicals quenching effect and modulating potential of bio transforming enzymes during DMBA exposed experimental mammary carcinogenesis. SUMMARY DMBA is a source of well-established site specific carcinogenHypnea musciformis act as a free radical

  8. Adenylate-forming enzymes

    Science.gov (United States)

    Schmelz, Stefan; Naismith, James H.

    2012-01-01

    Thioesters, amides and esters are common chemical building blocks in a wide array of natural products. The formation of these bonds can be catalyzed in a variety of ways. For chemists, the use of an activating group is a common strategy and adenylate enzymes are exemplars of this approach. Adenylating enzymes activate the otherwise unreactive carboxylic acid by transforming the normal hydroxyl leaving group into adenosine monophosphate. Recently there have been a number of studies of such enzymes and in this review we suggest a new classification scheme. The review highlights the diversity in enzyme fold, active site architecture and metal coordination that has evolved to catalyze this particular reaction. PMID:19836944

  9. Food and feed enzymes.

    Science.gov (United States)

    Fraatz, Marco Alexander; Rühl, Martin; Zorn, Holger

    2014-01-01

    Humans have benefited from the unique catalytic properties of enzymes, in particular for food production, for thousands of years. Prominent examples include the production of fermented alcoholic beverages, such as beer and wine, as well as bakery and dairy products. The chapter reviews the historic background of the development of modern enzyme technology and provides an overview of the industrial food and feed enzymes currently available on the world market. The chapter highlights enzyme applications for the improvement of resource efficiency, the biopreservation of food, and the treatment of food intolerances. Further topics address the improvement of food safety and food quality.

  10. Microbial amylolytic enzymes.

    Science.gov (United States)

    Vihinen, M; Mäntsälä, P

    1989-01-01

    Starch-degrading, amylolytic enzymes are widely distributed among microbes. Several activities are required to hydrolyze starch to its glucose units. These enzymes include alpha-amylase, beta-amylase, glucoamylase, alpha-glucosidase, pullulan-degrading enzymes, exoacting enzymes yielding alpha-type endproducts, and cyclodextrin glycosyltransferase. Properties of these enzymes vary and are somewhat linked to the environmental circumstances of the producing organisms. Features of the enzymes, their action patterns, physicochemical properties, occurrence, genetics, and results obtained from cloning of the genes are described. Among all the amylolytic enzymes, the genetics of alpha-amylase in Bacillus subtilis are best known. Alpha-Amylase production in B. subtilis is regulated by several genetic elements, many of which have synergistic effects. Genes encoding enzymes from all the amylolytic enzyme groups dealt with here have been cloned, and the sequences have been found to contain some highly conserved regions thought to be essential for their action and/or structure. Glucoamylase appears usually in several forms, which seem to be the results of a variety of mechanisms, including heterogeneous glycosylation, limited proteolysis, multiple modes of mRNA splicing, and the presence of several structural genes.

  11. Magnetically responsive enzyme powders

    Energy Technology Data Exchange (ETDEWEB)

    Pospiskova, Kristyna, E-mail: kristyna.pospiskova@upol.cz [Regional Centre of Advanced Technologies and Materials, Palacky University, Slechtitelu 11, 783 71 Olomouc (Czech Republic); Safarik, Ivo, E-mail: ivosaf@yahoo.com [Regional Centre of Advanced Technologies and Materials, Palacky University, Slechtitelu 11, 783 71 Olomouc (Czech Republic); Department of Nanobiotechnology, Institute of Nanobiology and Structural Biology of GCRC, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic)

    2015-04-15

    Powdered enzymes were transformed into their insoluble magnetic derivatives retaining their catalytic activity. Enzyme powders (e.g., trypsin and lipase) were suspended in various liquid media not allowing their solubilization (e.g., saturated ammonium sulfate and highly concentrated polyethylene glycol solutions, ethanol, methanol, 2-propanol) and subsequently cross-linked with glutaraldehyde. Magnetic modification was successfully performed at low temperature in a freezer (−20 °C) using magnetic iron oxides nano- and microparticles prepared by microwave-assisted synthesis from ferrous sulfate. Magnetized cross-linked enzyme powders were stable at least for two months in water suspension without leakage of fixed magnetic particles. Operational stability of magnetically responsive enzymes during eight repeated reaction cycles was generally without loss of enzyme activity. Separation of magnetically modified cross-linked powdered enzymes from reaction mixtures was significantly simplified due to their magnetic properties. - Highlights: • Cross-linked enzyme powders were prepared in various liquid media. • Insoluble enzymes were magnetized using iron oxides particles. • Magnetic iron oxides particles were prepared by microwave-assisted synthesis. • Magnetic modification was performed under low (freezing) temperature. • Cross-linked powdered trypsin and lipase can be used repeatedly for reaction.

  12. Artificial Enzymes, "Chemzymes"

    DEFF Research Database (Denmark)

    Bjerre, Jeannette; Rousseau, Cyril Andre Raphaël; Pedersen, Lavinia Georgeta M

    2008-01-01

    Enzymes have fascinated scientists since their discovery and, over some decades, one aim in organic chemistry has been the creation of molecules that mimic the active sites of enzymes and promote catalysis. Nevertheless, even today, there are relatively few examples of enzyme models that successf......Enzymes have fascinated scientists since their discovery and, over some decades, one aim in organic chemistry has been the creation of molecules that mimic the active sites of enzymes and promote catalysis. Nevertheless, even today, there are relatively few examples of enzyme models...... that successfully perform Michaelis-Menten catalysis under enzymatic conditions (i.e., aqueous medium, neutral pH, ambient temperature) and for those that do, very high rate accelerations are seldomly seen. This review will provide a brief summary of the recent developments in artificial enzymes, so called...... "Chemzymes", based on cyclodextrins and other molecules. Only the chemzymes that have shown enzyme-like activity that has been quantified by different methods will be mentioned. This review will summarize the work done in the field of artificial glycosidases, oxidases, epoxidases, and esterases, as well...

  13. Enzymes in Fermented Fish.

    Science.gov (United States)

    Giyatmi; Irianto, H E

    Fermented fish products are very popular particularly in Southeast Asian countries. These products have unique characteristics, especially in terms of aroma, flavor, and texture developing during fermentation process. Proteolytic enzymes have a main role in hydrolyzing protein into simpler compounds. Fermentation process of fish relies both on naturally occurring enzymes (in the muscle or the intestinal tract) as well as bacteria. Fermented fish products processed using the whole fish show a different characteristic compared to those prepared from headed and gutted fish. Endogenous enzymes like trypsin, chymotrypsin, elastase, and aminopeptidase are the most involved in the fermentation process. Muscle tissue enzymes like cathepsins, peptidases, transaminases, amidases, amino acid decarboxylases, glutamic dehydrogenases, and related enzymes may also play a role in fish fermentation. Due to the decreased bacterial number during fermentation, contribution of microbial enzymes to proteolysis may be expected prior to salting of fish. Commercial enzymes are supplemented during processing for specific purposes, such as quality improvement and process acceleration. In the case of fish sauce, efforts to accelerate fermentation process and to improve product quality have been studied by addition of enzymes such as papain, bromelain, trypsin, pepsin, and chymotrypsin. © 2017 Elsevier Inc. All rights reserved.

  14. An ultrastructural and histochemical study of the flexor tibialis muscle fiber types in male and female stick insects (Eurycantha calcarata, L).

    Science.gov (United States)

    Pilehvarian, Ali Asghar

    2015-10-01

    In this study the ultrastructural and histochemical characteristics of the flexor tibialis muscle fibers of the specialized metathoracic legs in the male and those of homologous and unspecialized ones in the female stick insects, Eurycantha calcarata, L, were examined. For the ultrastructural analysis, the muscle was divided longitudinally and vertically to produce a total of 12 sample parts e.g., anterior-dorsal-distal (ADD), posterior-ventral-medial (PVM) and so on. Light and electron microscopes were used to observe the muscle tissue. The methods for myosin adenosine triphosphatase (mATPase) and nicotine adenine dinucleotide- tetrazolium (NADH-TR) staining were modified from the methods of (Stokes et al., '79; Anttila et al., 2009; Anttila and Manttari, 2009). Sections with thickness of 22 μm, were cut from the anterior and the posterior surfaces of the muscle, using a cryostat. The histochemical and ultrastructural results showed that the muscles of both the male and the female were mixtures of physiological fiber types, with predominantly fast fibers. The muscles were composed of fibers with different staining properties for both mATPase and NADH-TR activities. The population of fibers within the muscles was heterogeneous. The differences between the population of the male and that of the female were significant. The means of most criteria e.g., mitochondrial amount and sarcoplasmic reticulum area predicted that the muscle of the male contained more fast fibers than the female. The histochemical examination also showed that the muscle of the male contained more fibers stained darkly for mATPase and lightly for NADH-TR.

  15. Effects of chloroquine on the adrenocortical function. II. Histological, histochemical and biochemical changes in the suprarenal gland of rats on long-term administration of chloroquine.

    Science.gov (United States)

    Grundmann, M; Bayer, A

    1976-01-01

    White female Wistar rats were used in order to study the influence of long-term oral application of 7-chloro-4-(4-diethylamino-1-methylbutylamino) quinoline (chloroquine) in doses of 30, 40 and 80 mg of base/kg upon the suprarenal gland. Histological, histochemical and biochemical findings give evidence of adrenocortical activation induced by chloroquine at all dose levels tested. The differences between the signs of adrenocortical activation as observed after the various doses were only those of quantity and time onset. The results indicate that the stimulation of the suprarenal cortex produced by repeated administration of chloroquine is not solely a manifestation of toxic action of chloroquine.

  16. Cotton cellulose: enzyme adsorption and enzymic hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Beltrame, P.L.; Carniti, P.; Focher, B.; Marzetti, A.; Cattaneo, M.

    1982-01-01

    The adsorption of a crude cellulase complex from Trichoderma viride on variously pretreated cotton cellulose samples was studied in the framework of the Langmuir approach at 2-8 degrees. The saturation amount of adsorbed enzyme was related to the susceptibility of the substrates to hydrolysis. In every case the adsorption process was faster by 2-3 orders of magnitude than the hydrolysis step to give end products. For ZnCl/sub 2/-treated cotton cellulose the Langmuir parameters correlated fairly well with the value of the Michaelis constant, measured for its enzymic hydrolysis, and the adsorptive complex was indistinguishable from the complex of the Michaelis-Menten model for the hydrolysis.

  17. On the Nature of Tintinnid Loricae (Ciliophora: Spirotricha: Tintinnina): a Histochemical, Enzymatic, EDX, and High-resolution TEM Study.

    Science.gov (United States)

    Agatha, Sabine; Simon, Paul

    2012-06-01

    , ultrastructure of the lorica forming granules, and the histochemical and enzymatic findings are not evident. Therefore, further studies are required to estimate the taxonomic significance of these features and the ecological role of sedimenting loricae.

  18. Comparative histochemical study of Bowen’s disease and actinic keratosis: preserved normal basal cells in Bowen’s disease

    Directory of Open Access Journals (Sweden)

    H Ishida

    2009-12-01

    Full Text Available The degree of DNA-instability as revealed by immunohistochemical staining with anti-cytidine antibody after acid hydrolysis (DNA-instability test has been recently used as a marker of malignancy. This technique was applied to examine 17 skin tissue samples of Bowen’s disease, 47 of actinic keratosis, 15 of squamous cell carcinoma, 5 of seborrheic keratosis, and 10 of normal skin. All benign neoplastic cells of seborrheic keratosis and normal epidermal cells were negative. On the other hand, all cancer cells were positive with the DNA-instability test, indicating their malignancy, but all basal cells in Bowen’s disease were completely negative. Compatible with this result, the basal cells in Bowen’s disease were characteristically normal as evident in other histochemical examinations. Thus, they were negative with p53 immunohistochemistry, with normal signals of chromosome 17 in situ hybridisation and argyrophilic nucleolar organiser region, and showed slightly enhanced proliferative activity as revealed by proliferating cell nuclear antigen immunohistochemistry. Immunohistochemical staining with 34 ß E12 (monoclonal antibody against cytokeratins 1, 5, 10, and 14, which stains all normal epidermal keratinocytes including basal cells, showed that only the basal cells of Bowen’s disease stained strongly and homogeneously, while all cancer cells in the upper layers of Bowen’s disease and all layers of actinic keratosis were only sporadically or weakly stained. Staining with 34 ß B4 (monoclonal antibody against cytokeratin 1, which recognises the whole epidermis except for the basal layer in the normal epidermis, showed that the basal cells in the Bowen’s disease were completely negative, and lower layer cells in the actinic keratosis and upper layer cells in Bowen’s disease were only sporadically stained positive, although the superficial layer cells in actinic keratosis stained strongly and homogeneously. Our findings clearly

  19. Characterization of the egg vesicular components in the seaweed, Fucus serratus L. (Fucales, Phaeophyta), using enzyme histochemistry and vital staining: the search for a lysosome-like body.

    Science.gov (United States)

    Holland, R D; Pitt, D; Moore, M N; Brownlee, C

    1997-03-01

    Fucus serratus eggs were examined for evidence of the existence of a lysosome-like body using enzyme histochemical and vital staining techniques. Simultaneous coupling azo-dye techniques for lysosomal acid phosphatase proved inappropriate owing to endogenous phenolic binding artefacts. The large number of alginate polysaccharide and polyphenolic egg vesicles interfered with vital staining techniques for lysosomes. Lysosomal esterase activity was detected in the abundant egg lipid bodies. The role of the egg lipid body as an equivalent lysosome-like body of higher plants, the spherosome, is discussed in relation to egg fertilization and early zygote development.

  20. The EBI enzyme portal.

    Science.gov (United States)

    Alcántara, Rafael; Onwubiko, Joseph; Cao, Hong; Matos, Paula de; Cham, Jennifer A; Jacobsen, Jules; Holliday, Gemma L; Fischer, Julia D; Rahman, Syed Asad; Jassal, Bijay; Goujon, Mikael; Rowland, Francis; Velankar, Sameer; López, Rodrigo; Overington, John P; Kleywegt, Gerard J; Hermjakob, Henning; O'Donovan, Claire; Martín, María Jesús; Thornton, Janet M; Steinbeck, Christoph

    2013-01-01

    The availability of comprehensive information about enzymes plays an important role in answering questions relevant to interdisciplinary fields such as biochemistry, enzymology, biofuels, bioengineering and drug discovery. At the EMBL European Bioinformatics Institute, we have developed an enzyme portal (http://www.ebi.ac.uk/enzymeportal) to provide this wealth of information on enzymes from multiple in-house resources addressing particular data classes: protein sequence and structure, reactions, pathways and small molecules. The fact that these data reside in separate databases makes information discovery cumbersome. The main goal of the portal is to simplify this process for end users.

  1. Enzyme molecules as nanomotors.

    Science.gov (United States)

    Sengupta, Samudra; Dey, Krishna K; Muddana, Hari S; Tabouillot, Tristan; Ibele, Michael E; Butler, Peter J; Sen, Ayusman

    2013-01-30

    Using fluorescence correlation spectroscopy, we show that the diffusive movements of catalase enzyme molecules increase in the presence of the substrate, hydrogen peroxide, in a concentration-dependent manner. Employing a microfluidic device to generate a substrate concentration gradient, we show that both catalase and urease enzyme molecules spread toward areas of higher substrate concentration, a form of chemotaxis at the molecular scale. Using glucose oxidase and glucose to generate a hydrogen peroxide gradient, we induce the migration of catalase toward glucose oxidase, thereby showing that chemically interconnected enzymes can be drawn together.

  2. Histochemical investigations on the in vivo effects of fluoride on tricarboxylic acid cycle dehydrogenases from Pelargonium zonale. Part II

    Energy Technology Data Exchange (ETDEWEB)

    Lovelace, C.J.; Miller, G.W.

    1967-01-01

    In vivo effects of fluoride on tricarboxylic acid (TCA) cycle dehydrogenase enzymes of Pelargonium zonale were studied using p-nitro blue tetrazoleum chloride. Plants were exposed to 17 ppb HF, and enzyme activities in treated plants were compared to those in controls. Leaves of control plants were incubated in 5 x 10/sup -3/ M sodium fluoride. Injuries observed in fumigation and solution experiments were similar. Leaf tissue subjected to HF or sodium fluoride evidenced less succinic p-nitro blue tetrazoleum reductase activity than did control tissue. Other TCA cycle dehydrogenase enzymes were not observably affected by the fluoride concentrations used in these experiments. Excised leaves cultured in 5 x 10/sup -3/ M sodium fluoride exhibited less succinic p-nitro blue tetrazoleum reductase activity after 24 hr than did leaves cultured in 5 x 10/sup -3/ M sodium chloride. 8 references, 8 figures.

  3. Histochemical studies on intestinal metaplasia adjacent to gastric cardia adenocarcinoma in subjects at high-incidence area in Henan, north China

    Institute of Scientific and Technical Information of China (English)

    She-Gan Gao; Hua-Qin Guo; Jun-Kuan Wang; Xiao-Shan Feng; Bao-Gen Ma; Li-Dong Wang; Zong-Min Fan; Ji-Lin Li; Xin He; Rui-Feng Guo; Dong-Ling Xie; Xin-Wei He; Shan-Shan Gao

    2005-01-01

    AIM: To characterize the histochemical type and pattern of intestinal metaplasia (IM) adjacent to gastric cardia adenocarcinoma (GCA) and distal gastric cancer (GC) in Linzhou, Henan Province, China.METHODS: Alcian-blue-periodic acid Schiff and high iron diamine-Alcian blue histochemical methods were performed on 142 cases of IM, including 49 cases of GCA and 93 cases of GC. All the patients were from Linzhou, Henan Province, China, the highest incidence area for both GCA and squamous cell carcinoma. Radio- or chemotherapy was not applied to these patients before surgery.RESULTS: The detection rate of IM in tissues adjacent to GCA tissues was 44.9%, which was significantly lower than that in GC tissues (80.64%, P<0.01). The rates of both incomplete small intestinal and colonic IM types identified by histochemistry in GCA tissues (31.82% and 63.64%, respectively) were significantly higher than those in GC (5.33% and 21.33%, respectively, P<0.01).CONCLUSION: IM in GCA and GC should be considered as a separate entity. Further research is needed to evaluate whether neoplastic progression of IM is related to its mucin profile in GCA.

  4. Comparison of immunohistochemical, histochemical and immunochemical methods for the detection of wheat protein allergens in meat samples and cooked, dry, raw and fermented sausage samples.

    Science.gov (United States)

    Lukášková, Z Řezáčová; Tremlová, B; Pospiech, M; Renčová, E; Randulová, Z; Steinhauser, L; Reichová, A; Bednář, J

    2011-01-01

    Nowadays is it a common practice to add vegetable protein in the production of meat products. Because of the possible substitution of high-quality raw meat with vegetable protein without the labelling the product package and because of the allergenic potential of many vegetable proteins, it is important to develop accurate methods for its detection. The objective of the study was to compare histochemical, immunochemical (ELISA, ALERT gliadin screening test) and immunohistochemical methods for the detection of wheat protein in meat samples and sausages. Histochemical methods were useful for the detection of flour in meat samples, but the immunohistochemical method was better for the detection of wheat protein. ALERT gliadin screening test detected gliadin from 10 mg kg(-1), while an immunohistochemical method detected wheat protein concentrations from 1 g kg(-1) and an ELISA method detected wheat protein concentrations from 4 g kg(-1). ALERT gliadin screening test showed results within 1 day, whilst an ELISA detection method took 2 days, and an immunohistochemical procedure took 5 days at the soonest, all including sample preparation. This study also focused on optimisation of an immunohistochemical method for samples of cooked sausage. In addition, three samples were sufficient for wheat protein detection at a concentration of 1 g kg(-1) (and greater) with a confidence level greater than 95%.

  5. Enzymes in Analytical Chemistry.

    Science.gov (United States)

    Fishman, Myer M.

    1980-01-01

    Presents tabular information concerning recent research in the field of enzymes in analytic chemistry, with methods, substrate or reaction catalyzed, assay, comments and references listed. The table refers to 128 references. Also listed are 13 general citations. (CS)

  6. Enzymic lactose hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Miller, J.J.; Brand, J.C.

    1980-01-01

    Acid or enzymic hydrolysis can be used to hydrolyze lactose. Advantages of both are compared and details of enzymic hydrolysis using yeast or fungal enzymes given. The new scheme outlined involves recycling lactase. Because lactose and lactase react to ultrafiltration (UF) membranes differently separation is possible. Milk or milk products are ultrafiltered to separate a concentrate from a lactose-rich permeate which is treated with lactase in a reactor until hydrolysis reaches a required level. The lactase can be removed by UF as it does not permeate the membrane, and it is recycled back to the reactor. Permeate from the second UF stage may or may not be recombined with the concentrate from the first stage to produce a low lactose product (analysis of a typical low-lactose dried whole milk is given). Batch or continuous processes are explained and a batch process without enzyme recovery is discussed. (Refs. 4).

  7. Membrane Assisted Enzyme Fractionation

    DEFF Research Database (Denmark)

    Yuan, Linfeng

    . In this thesis, separations using crossflow elecro-membrane filtration (EMF) of amino acids, bovine serum albumin (BSA) and industrial enzymes from Novozymes were performed. The main objective of this study was to investigate the technological feasibility of EMF in the application of industrial enzyme...... fractionation, such as removal of a side activity from the main enzyme activity. As a proof-of-concept, amino acids were used as model solution to test the feasibility of EMF in the application of amphoteric molecule separation. A single amino acid was used to illustrate the effect of an electric field...... on the separation performance were very small in the investigated range. The mass transport of each enzyme can be well explained by the Extended-Nernst-Planck equation. Better separation was observed at lower feed concentration, higher solution pH in the investigated range and with a polysulfone (PS) MF membrane...

  8. Indicators: Sediment Enzymes

    Science.gov (United States)

    Sediment enzymes are proteins that are produced by microorganisms living in the sediment or soil. They are indicators of key ecosystem processes and can help determine which nutrients are affecting the biological community of a waterbody.

  9. Starch Biorefinery Enzymes.

    Science.gov (United States)

    Läufer, Albrecht

    2017-03-07

    Nature uses enzymes to build and convert biomass; mankind uses the same enzymes and produces them on a large scale to make optimum use of biomass in biorefineries. Bacterial α-amylases and fungal glucoamylases have been the workhorses of starch biorefineries for many decades. Pullulanases were introduced in the 1980s. Proteases, cellulases, hemicellulases, and phytases have been on the market for a few years as process aids, improving yields, performance, and costs. Detailed studies of the complex chemical structures of biomass and of the physicochemical limitations of industrial biorefineries have led enzyme developers to produce novel tailor-made solutions for improving yield and profitability in the industry. This chapter reviews the development of enzyme applications in the major starch biorefining processes.

  10. Membrane Assisted Enzyme Fractionation

    DEFF Research Database (Denmark)

    Yuan, Linfeng

    . In this thesis, separations using crossflow elecro-membrane filtration (EMF) of amino acids, bovine serum albumin (BSA) and industrial enzymes from Novozymes were performed. The main objective of this study was to investigate the technological feasibility of EMF in the application of industrial enzyme...... fractionation, such as removal of a side activity from the main enzyme activity. As a proof-of-concept, amino acids were used as model solution to test the feasibility of EMF in the application of amphoteric molecule separation. A single amino acid was used to illustrate the effect of an electric field...... on the separation performance were very small in the investigated range. The mass transport of each enzyme can be well explained by the Extended-Nernst-Planck equation. Better separation was observed at lower feed concentration, higher solution pH in the investigated range and with a polysulfone (PS) MF membrane...

  11. [Detection of enzyme activity in decontaminated spices in industrial use].

    Science.gov (United States)

    Müller, R; Theobald, R

    1995-03-01

    A range of decontaminated species of industrial use have been examined for their enzymes (catalase, peroxidase, amylase, lipase activity). The genuine enzymes remain fully active in irradiated spices, whereas the microbial load is clearly reduced. In contrast steam treated spices no longer demonstrate enzyme activities. Steam treatment offers e.g. black pepper without lipase activity, which can no longer cause fat deterioration. Low microbial load in combination with clearly detectable enzyme activity in spices is an indication for irradiation, whereas, reduced microbial contamination combined with enzyme inactivation indicate steam treatment of raw material.

  12. RNA-modifying enzymes.

    Science.gov (United States)

    Ferré-D'Amaré, Adrian R

    2003-02-01

    A bewildering number of post-transcriptional modifications are introduced into cellular RNAs by enzymes that are often conserved among archaea, bacteria and eukaryotes. The modifications range from those with well-understood functions, such as tRNA aminoacylation, to widespread but more mysterious ones, such as pseudouridylation. Recent structure determinations have included two types of RNA nucleobase modifying enzyme: pseudouridine synthases and tRNA guanine transglycosylases.

  13. Overproduction of ligninolytic enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Elisashvili, Vladimir; Kachlishvili, Eva; Torok, Tamas

    2014-06-17

    Methods, compositions, and systems for overproducing ligninolytic enzymes from the basidiomycetous fungus are described herein. As described, the method can include incubating a fungal strain of Cerrena unicolor IBB 303 in a fermentation system having growth medium which includes lignocellulosic material and then cultivating the fungal strain in the fermentation system under conditions wherein the fungus expresses the ligninolytic enzymes. In some cases, the lignocellulosic material is mandarin peel, ethanol production residue, walnut pericarp, wheat bran, wheat straw, or banana peel.

  14. [Involution of the mammary gland. Enzyme histochemistry, elektron microscopy and radioautography (author's transl)].

    Science.gov (United States)

    Korfsmeier, K

    1976-01-01

    A study has been made of the progress of involution of the mouse and rat mammary gland using histologic, electron microscopic, histochemical and autoradiographic methods. Particular emphasis has been placed on the morphology, metabolic alterations and activities of histochemically identifiable enzymes, and on the pharmacologic effects of lactation inhibiting agents and cytostatic drugs on lactation and involution. In order to allow a systematic investigation, involution was initiated in rats and mice by ligation of individual gland ducts at various time intervals. Both lactating glands and glands in different phases of involution were thus available in a given animal. The most important observation was that involution, which altogether takes approximately 2 weeks to be complete, involves a three-phase process, each phase being clearly distinguishable by morphologic and histochemical criteria. The first phase comprises approximately 4 days during which production of milk may be reinitiated. The second phase starts on day 5 of involution and constitutes the period of involution per se characterized by appreciable parenchymal cell degradation. The third phase, which starts around day 10, is the period of reorganization to the resting mammary gland. Early in the first phase of involution, substantial alveolar enlargement due to engorgement with milk, together with epithelial flattening, are prominent features. By day 3, the glandular contents decrease again in volume, the number of glandular cells and the constituent cytoplasmic organelles remaining unchanged during this period, except for the diminished appearance of fat droplets. In addition to normal appearing vacuoles with only occasional or sparse protein granules, giant vacuoles containing, in part, several hundred casein granules are found. Their formation appears to be due to increased stacking of granules in distended vacuoles prior to dissociation from the Golgi apparatus. In addition, however, the enhanced

  15. Histochemical, Biochemical and Cell Biological aspects of tail regeneration in lizard, an amniote model for studies on tissue regeneration.

    Science.gov (United States)

    Alibardi, Lorenzo

    2014-01-01

    The present review summarizes biochemical, histochemical and immunocytochemical aspects of the process of tissue regeneration in lizards, non-mammalian amniotes with high regenerative power. The amputated tail initially mobilizes the glycogen and lipid reserves during wound healing. In the following stage of formation of the regenerative blastema tissue remodeling produces a typical embryonic tissue, initially increasing the amount of water and glycosaminoglycans such as jaluronate, which are later replaced by sulfated glycosaminoglycans and collagen during tail elongation. In blastematic and early differentiating stages the initial anaerobic metabolism utilizes glycolysis and hexose monophosphate pathways to sustain high RNA production and lipid catabolism for energy production. This stage, after formation of blood vessels, is replaced by the energy-efficient aerobic metabolism based on the Krebs' cycle that is needed for the differentiation and growth of the new tissues of the regenerating tail. Specific proteins of the cytoskeleton, extracellular matrix, cell junctions, transcriptional and growth factors are actively produced in the embryonic environment of early stages of regeneration and allow for cell movement, signaling and differentiation. During wound healing, the production of anti-microbial peptides in granulocytes is likely involved in limiting inflammation and stimulates tissue regeneration in the tail while the lasting inflammatory reaction of the limb and spinal cord limits their potential of regeneration. Activated hemopoiesis, circulating blood, endocrine glands, liver, kidney and spleen supply the regenerating tissues with metabolites and hormones but also with phagocytes and immuno-competent cells that can inhibit tissue regeneration after repetitive amputations that elicit chronic inflammation. The latter aspect shows how successful tissue regeneration in an amniote can be turned into scarring by the alteration of the initial microenvironment

  16. Enzyme replacement therapy for alpha-mannosidosis

    DEFF Research Database (Denmark)

    Borgwardt, Line Gutte; Dali, Christine I.; Fogh, J

    2013-01-01

    Alpha-mannosidosis (OMIM 248500) is a rare lysosomal storage disease (LSD) caused by alpha-mannosidase deficiency. Manifestations include intellectual disabilities, facial characteristics and hearing impairment. A recombinant human alpha-mannosidase (rhLAMAN) has been developed for weekly intrave...... intravenous enzyme replacement therapy (ERT). We present the preliminary data after 12 months of treatment....

  17. Random-walk enzymes.

    Science.gov (United States)

    Mak, Chi H; Pham, Phuong; Afif, Samir A; Goodman, Myron F

    2015-09-01

    Enzymes that rely on random walk to search for substrate targets in a heterogeneously dispersed medium can leave behind complex spatial profiles of their catalyzed conversions. The catalytic signatures of these random-walk enzymes are the result of two coupled stochastic processes: scanning and catalysis. Here we develop analytical models to understand the conversion profiles produced by these enzymes, comparing an intrusive model, in which scanning and catalysis are tightly coupled, against a loosely coupled passive model. Diagrammatic theory and path-integral solutions of these models revealed clearly distinct predictions. Comparison to experimental data from catalyzed deaminations deposited on single-stranded DNA by the enzyme activation-induced deoxycytidine deaminase (AID) demonstrates that catalysis and diffusion are strongly intertwined, where the chemical conversions give rise to new stochastic trajectories that were absent if the substrate DNA was homogeneous. The C→U deamination profiles in both analytical predictions and experiments exhibit a strong contextual dependence, where the conversion rate of each target site is strongly contingent on the identities of other surrounding targets, with the intrusive model showing an excellent fit to the data. These methods can be applied to deduce sequence-dependent catalytic signatures of other DNA modification enzymes, with potential applications to cancer, gene regulation, and epigenetics.

  18. Random-walk enzymes

    Science.gov (United States)

    Mak, Chi H.; Pham, Phuong; Afif, Samir A.; Goodman, Myron F.

    2015-09-01

    Enzymes that rely on random walk to search for substrate targets in a heterogeneously dispersed medium can leave behind complex spatial profiles of their catalyzed conversions. The catalytic signatures of these random-walk enzymes are the result of two coupled stochastic processes: scanning and catalysis. Here we develop analytical models to understand the conversion profiles produced by these enzymes, comparing an intrusive model, in which scanning and catalysis are tightly coupled, against a loosely coupled passive model. Diagrammatic theory and path-integral solutions of these models revealed clearly distinct predictions. Comparison to experimental data from catalyzed deaminations deposited on single-stranded DNA by the enzyme activation-induced deoxycytidine deaminase (AID) demonstrates that catalysis and diffusion are strongly intertwined, where the chemical conversions give rise to new stochastic trajectories that were absent if the substrate DNA was homogeneous. The C →U deamination profiles in both analytical predictions and experiments exhibit a strong contextual dependence, where the conversion rate of each target site is strongly contingent on the identities of other surrounding targets, with the intrusive model showing an excellent fit to the data. These methods can be applied to deduce sequence-dependent catalytic signatures of other DNA modification enzymes, with potential applications to cancer, gene regulation, and epigenetics.

  19. Random-walk enzymes

    Science.gov (United States)

    Mak, Chi H.; Pham, Phuong; Afif, Samir A.; Goodman, Myron F.

    2015-01-01

    Enzymes that rely on random walk to search for substrate targets in a heterogeneously dispersed medium can leave behind complex spatial profiles of their catalyzed conversions. The catalytic signatures of these random-walk enzymes are the result of two coupled stochastic processes: scanning and catalysis. Here we develop analytical models to understand the conversion profiles produced by these enzymes, comparing an intrusive model, in which scanning and catalysis are tightly coupled, against a loosely coupled passive model. Diagrammatic theory and path-integral solutions of these models revealed clearly distinct predictions. Comparison to experimental data from catalyzed deaminations deposited on single-stranded DNA by the enzyme activation-induced deoxycytidine deaminase (AID) demonstrates that catalysis and diffusion are strongly intertwined, where the chemical conversions give rise to new stochastic trajectories that were absent if the substrate DNA was homogeneous. The C → U deamination profiles in both analytical predictions and experiments exhibit a strong contextual dependence, where the conversion rate of each target site is strongly contingent on the identities of other surrounding targets, with the intrusive model showing an excellent fit to the data. These methods can be applied to deduce sequence-dependent catalytic signatures of other DNA modification enzymes, with potential applications to cancer, gene regulation, and epigenetics. PMID:26465508

  20. Endocarditis caused by Haemophilus aegyptius.

    Science.gov (United States)

    Porath, A; Wanderman, K; Simu, A; Vidne, B; Alkan, M

    1986-08-01

    Endocarditis due to Haemophilus is uncommon. This is the first reported case caused by Haemophilus aegyptius. The course of the disease was complicated by pericarditis, congestive heart failure, and myocardial abscess formation. Surgical removal of the damaged aortic valve was not beneficial. The biologic properties of the organism included urea degradation, absence of indole metabolism, and absence of the enzyme ornithine decarboxylase.

  1. Enzyme recycling in lignocellulosic biorefineries

    DEFF Research Database (Denmark)

    Jørgensen, Henning; Pinelo, Manuel

    2017-01-01

    platform. Cellulases are the most important enzymes required in this process, but the complex nature of lignocellulose requires several other enzymes (hemicellulases and auxiliary enzymes) for efficient hydrolysis. Enzyme recycling increases the catalytic productivity of the enzymes by reusing them...... upscaled and tested in industrial settings, mainly because of many difficulties with recycling of enzymes from the complex lignocellulose hydrolyzate at industrially relevant conditions, i.e., high solids loadings. The challenges are associated with the large number of different enzymes required...... for efficient hydrolysis, enzyme stability, and the detrimental interaction between enzyme and lignin. This review provides a comprehensive overview of the various methods for enzyme recovery and recycling, for example recycling of free enzymes, readsorption to fresh material, recycling of solids, membrane...

  2. Evolution of enzymes in metabolism: a network perspective.

    Science.gov (United States)

    Alves, Rui; Chaleil, Raphael A G; Sternberg, Michael J E

    2002-07-19

    Several models have been proposed to explain the origin and evolution of enzymes in metabolic pathways. Initially, the retro-evolution model proposed that, as enzymes at the end of pathways depleted their substrates in the primordial soup, there was a pressure for earlier enzymes in pathways to be created, using the later ones as initial template, in order to replenish the pools of depleted metabolites. Later, the recruitment model proposed that initial templates from other pathways could be used as long as those enzymes were similar in chemistry or substrate specificity. These two models have dominated recent studies of enzyme evolution. These studies are constrained by either the small scale of the study or the artificial restrictions imposed by pathway definitions. Here, a network approach is used to study enzyme evolution in fully sequenced genomes, thus removing both constraints. We find that homologous pairs of enzymes are roughly twice as likely to have evolved from enzymes that are less than three steps away from each other in the reaction network than pairs of non-homologous enzymes. These results, together with the conservation of the type of chemical reaction catalyzed by evolutionarily related enzymes, suggest that functional blocks of similar chemistry have evolved within metabolic networks. One possible explanation for these observations is that this local evolution phenomenon is likely to cause less global physiological disruptions in metabolism than evolution of enzymes from other enzymes that are distant from them in the metabolic network.

  3. Entropy and Enzyme Catalysis.

    Science.gov (United States)

    Åqvist, Johan; Kazemi, Masoud; Isaksen, Geir Villy; Brandsdal, Bjørn Olav

    2017-02-21

    The role played by entropy for the enormous rate enhancement achieved by enzymes has been debated for many decades. There are, for example, several confirmed cases where the activation free energy is reduced by around 10 kcal/mol due to entropic effects, corresponding to a rate enhancement of ∼10(7) compared to the uncatalyzed reaction. However, despite substantial efforts from both the experimental and theoretical side, no real consensus has been reached regarding the origin of such large entropic contributions to enzyme catalysis. Another remarkable instance of entropic effects is found in enzymes that are adapted by evolution to work at low temperatures, near the freezing point of water. These cold-adapted enzymes invariably show a more negative entropy and a lower enthalpy of activation than their mesophilic orthologs, which counteracts the exponential damping of reaction rates at lower temperature. The structural origin of this universal phenomenon has, however, remained elusive. The basic problem with connecting macroscopic thermodynamic quantities, such as activation entropy and enthalpy derived from Arrhenius plots, to the 3D protein structure is that the underlying detailed (microscopic) energetics is essentially inaccessible to experiment. Moreover, attempts to calculate entropy contributions by computer simulations have mostly focused only on substrate entropies, which do not provide the full picture. We have recently devised a new approach for accessing thermodynamic activation parameters of both enzyme and solution reactions from computer simulations, which turns out to be very successful. This method is analogous to the experimental Arrhenius plots and directly evaluates the temperature dependence of calculated reaction free energy profiles. Hence, by extensive molecular dynamics simulations and calculations of up to thousands of independent free energy profiles, we are able to extract activation parameters with sufficient precision for making

  4. Angiotensin-converting enzyme

    DEFF Research Database (Denmark)

    Sørensen, P G; Rømer, F K; Cortes, D

    1984-01-01

    In order to evaluate bleomycin-associated lung damage in humans, lung function parameters and serum levels of the endothelial-bound angiotensin-converting enzyme (ACE) were determined by serial measurements in 11 patients who were treated for testicular cancer. None developed clinical or radiolog......In order to evaluate bleomycin-associated lung damage in humans, lung function parameters and serum levels of the endothelial-bound angiotensin-converting enzyme (ACE) were determined by serial measurements in 11 patients who were treated for testicular cancer. None developed clinical...

  5. SH groups in the alpha-naphthyl acetate esterase in the thyroid of the guinea-pig. A histochemical study

    DEFF Research Database (Denmark)

    Kirkeby, S

    1976-01-01

    The alpha-naphthyl acetate esterase in both group I and group II thyroid cells is shown to contain SH groups since there is a decline in activity in both cell groups when certain sulfhydryl reagents [DTNB; 5,5'-Dithiobis-(2-nitrobenzoic acid)-AgNO3-Mersalyl-PCMB (parachloro mercuribenzoate) + ure...... could only inhibit the activity of the alpha-naphthylacetate esterase if the enzyme was denaturated with 5 M urea. The maximal inhibitory effect of PCMB was only obtained when NaCl was added to the incubation media. The most effective inhibitor was AgNO3....... if cysteine was added to the incubation media in equivalent concentrations to the SH blocker. There were great differences among the sulfhydryl reagents used in their ability to bring about enzyme inhibition. The alkylating agents NEM (N-ethylmaleimide) and iodoacetamide had no or little effect while PCMB...

  6. Purification of highly chlorinated dioxins degrading enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Ishii, K.; Furuichi, T.; Koike, K.; Kuboshima, M. [Hokkaido Univ. (Japan). Division of Environment Resource Engineering, Graduate School of Engineering

    2004-09-15

    Soil contamination caused by dioxins in and around sites of incinerators for municipal solid waste (MSW) is a concern in Japan. For example, scattering wastewater from a wet gas scrubber at an MSW incinerator facility in Nose, Osaka caused soil and surface water contamination. The concentration of dioxins in the soil was about 8,000 pg-TEQ/g. Other contamination sites include soils on which fly ash has been placed directly or improperly stored and landfill sites that have received bottom and fly ash over a long period. Some countermeasures are required immediately at these dioxins-contaminated sites. We have previously developed bioreactor systems for dioxin-contaminated water and soil. We have shown that a fungus, Pseudallescheria boydii (P. boydii), isolated from activated sludge treating wastewater that contained dioxins, has the ability to degrade highly chlorinated dioxins. A reaction product of octachlorinated dibenzo-p-dioxin (OCDD) was identified as heptachlorinated dibenzo-p-dioxin. Therefore, one of the pathways for degradation of OCDD by this fungus was predicted to be as follows: OCDD is transformed by dechlorination and then one of the remaining aromatic rings is oxidized. To apply P. boydii to on-site technologies (e.g., bioreactor systems), as well as in situ technologies, enzyme treatment using a dioxin-degrading enzyme from P. boydii needs to be developed because P. boydii is a weak pathogenic fungus, known to cause opportunistic infection. As a result, we have studied enzyme purification of nonchlorinated dioxin, namely, dibenzo-pdioxin (DD). However, we did not try to identify enzymes capable of degrading highly chlorinated dioxins. This study has elucidated a method of enzyme assay for measuring OCDD-degrading activity, and has attempted to purify OCDD-degrading enzymes from P. boydii using enzyme assay. In addition, as first step toward purifying 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), 2,3,7,8-TCDD degradation tests were carried out

  7. The surface science of enzymes

    DEFF Research Database (Denmark)

    Rod, Thomas Holm; Nørskov, Jens Kehlet

    2002-01-01

    One of the largest challenges to science in the coming years is to find the relation between enzyme structure and function. Can we predict which reactions an enzyme catalyzes from knowledge of its structure-or from its amino acid sequence? Can we use that knowledge to modify enzyme function......? To solve these problems we must understand in some detail how enzymes interact with reactants from its surroundings. These interactions take place at the surface of the enzyme and the question of enzyme function can be viewed as the surface science of enzymes. In this article we discuss how to describe...... catalysis by enzymes, and in particular the analogies between enzyme catalyzed reactions and surface catalyzed reactions. We do this by discussing two concrete examples of reactions catalyzed both in nature (by enzymes) and in industrial reactors (by inorganic materials), and show that although analogies...

  8. Amperometric Enzyme Electrodes

    Science.gov (United States)

    1989-12-01

    form of carbon (glascy carbon, graphite, reticulated vitreous carbon, carbon paste, fiber or foil). Carbon is favored for enzyme immoblization...interference from spurious electroactive species in blood, t proprietary multilayer membranie that includes a cellulose acetate memirane and a Nucleopore

  9. ISFET based enzyme sensors

    NARCIS (Netherlands)

    van der Schoot, Bart H.; Bergveld, Piet

    1987-01-01

    This paper reviews the results that have been reported on ISFET based enzyme sensors. The most important improvement that results from the application of ISFETs instead of glass membrane electrodes is in the method of fabrication. Problems with regard to the pH dependence of the response and the

  10. A lectin histochemical study on carbohydrate moieties of the gonadotropin-like substance in the epithelial cells of Hatschek's pit of Branchiostoma belcheri

    Science.gov (United States)

    Fang, Y. Q.; Welsch, U.

    1997-03-01

    The present light microscopic lectin, histochemical study suggests for the first time that the vertebrate gonadotropin-like substance in the basal part of the epithelial cells of Hatschek's pit is a sialic acid-containing glycoprotein. The binding intensity of the epithelial cells in Hatschek's pit to 6 lectins ( Limulus polyphemus agglutinin (LPA), Wheat germ agglutinin (WGA), Helix pomatia agglutinin (HPA), Concanavalin A (Con A), Ulex europaeus agglutinin I (UEA I) and Ricinus communis agglutinin I (RCA I)) indicate that the carbohydrate composition of the gonadotrophic glycoprotein is similar to that of mammals and fish, and that N-acetyl-D-galactosamine, sialic acid, glucosamine, D-mannose and L-fucose are components of the carbohydrate portion.

  11. Expression cloning of different bacterial phosphatase-encoding genes by histochemical screening of genomic libraries onto an indicator medium containing phenolphthalein diphosphate and methyl green.

    Science.gov (United States)

    Riccio, M L; Rossolini, G M; Lombardi, G; Chiesurin, A; Satta, G

    1997-02-01

    A system for expression cloning of bacterial phosphatase-encoding genes has been developed, and its potential has been investigated. The system is based on histochemical screening of bacterial genomic libraries, constructed in an Escherichia coli multicopy plasmid vector, for phosphatase-producing clones using an indicator medium (named TPMG) made of Tryptose-Phosphate agar supplemented with the phosphatase substrate phenolphthalein diphosphate and the stain methyl green. To test the performance of this system, three genomic libraries were constructed from bacterial strains of different species which showed different patterns of phosphatase activity, and were screened using the TPMG medium. Following a partial screening, three different phosphatase-encoding genes (respectively encoding a class A non-specific acid phosphatase, an acid-hexose phosphatase and a non-specific alkaline phosphatase) were shotgun-cloned from the above libraries, indicating that the TPMG-based expression cloning system can be useful for rapid isolation of different bacterial phosphatase-encoding genes.

  12. Computational enzyme design

    Science.gov (United States)

    Bolon, Daniel N.

    2002-08-01

    The long-term objective of computational enzyme design is the ability to generate efficient protein catalysts for any chemical reaction. This thesis develops and experimentally validates a general computational approach for the design of enzymes with novel function. In order to include catalytic mechanism in protein design, a high-energy state (HES) rotamer (side chain representation) was constructed. In this rotamer, substrate atoms are in a HES. In addition, at least one amino acid side chain is positioned to interact favorably with substrate atoms in their HES and facilitate the reaction. Including an amino acid side chain in the HES rotamer automatically positions substrate relative to a protein scaffold and allows protein design algorithms to search for sequences capable of interacting favorably with the substrate. Because chemical similarity exists between the transition state and the high-energy state, optimizing the protein sequence to interact favorably with the HES rotamer should lead to transition state stabilization. In addition, the HES rotamer model focuses the subsequent computational active site design on a relevant phase space where an amino acid is capable of interacting in a catalytically active geometry with substrate. Using a HES rotamer model of the histidine mediated nucleophilic hydrolysis of p-nitrophenyl acetate, the catalytically inert 108 residue E. coli thioredoxin as a scaffold, and the ORBIT protein design software to compute sequences, an active site scan identified two promising active site designs. Experimentally, both candidate ?protozymes? demonstrated catalytic activity significantly above background. In addition, the rate enhancement of one of these ?protozymes? was the same order of magnitude as the first catalytic antibodies. Because polar groups are frequently buried at enzyme-substrate interfaces, improved modeling of buried polar interactions may benefit enzyme design. By studying native protein structures, rules have been

  13. Protective Effects of Royal Jelly and Vitamin C against Experimental Hemolytic Anemia on Sex Hormones and Histochemical Testicle Tissue Histochemistry of Adult Mice

    Directory of Open Access Journals (Sweden)

    H Anbara

    2016-03-01

    Full Text Available Introduction: Phenylhydrazine (PHZ is a well-known hemolytic compound inducing intoxication in erythrocytes. Therefore, the present study aimed to evaluate the protective effects of royal jelly and vitamin C against phenylhydrazine-induced damages in mouse testicles. Methods: In this study, 64 adult male mice were randomly and equally assigned to eight groups. The first group received normal saline (0.1ml intraperitoneally. The second group received PHZ (6 mg/100 gr intraperitoneally in 48-hour intervals. The third group received vitamin C (250 mg/kg/day intraperitoneally perday a long with PHZ. The fourth group received royal jelly (100 mg/kg/day through gavage. The fifth group received PHZ along with vitamin C and royal jelly in similar doses to the previous groups. The sixth group received only vitamin C, the seventh group recieved only royal jelly, and finally the eighth group received similar doses of vitamin C and royal jelly. After 35 days, serum and tissue samples were taken and used for histochemical (Mallory-Azan, Alkaline phosphatase, Oil red-O and PAS, and serum analyses (Testosterone, LH, FSH. Results: The study results revealed the histochemical changes in testicular tissue of the phenylhydrazine group, in which vitamin C and royal jelly partly improved the changes. Furthermore, serum analyses demonstrated a significant decrease in testosterone, FSH and LH levels, which this decrease was diminished by royal jelly and vitamin C. Conclusions: Royal jelly and vitamin C seem to have the potential to decrease serum and tissue damages induced by phenylhydrazine via restraining free radicals.

  14. The Moderately Efficient Enzyme: Futile Encounters and Enzyme Floppiness.

    Science.gov (United States)

    Bar-Even, Arren; Milo, Ron; Noor, Elad; Tawfik, Dan S

    2015-08-18

    The pioneering model of Henri, Michaelis, and Menten was based on the fast equilibrium assumption: the substrate binds its enzyme reversibly, and substrate dissociation is much faster than product formation. Here, we examine this assumption from a somewhat different point of view, asking what fraction of enzyme-substrate complexes are futile, i.e., result in dissociation rather than product formation. In Knowles' notion of a "perfect" enzyme, all encounters of the enzyme with its substrate result in conversion to product. Thus, the perfect enzyme's catalytic efficiency, kcat/KM, is constrained by only the diffusion on-rate, and the fraction of futile encounters (defined as φ) approaches zero. The available data on >1000 different enzymes suggest that for ≥90% of enzymes φ > 0.99 and for the "average enzyme" φ ≥ 0.9999; namely, <1 of 10(4) encounters is productive. Thus, the "fast equilibrium" assumption holds for the vast majority of enzymes. We discuss possible molecular origins for the dominance of futile encounters, including the coexistence of multiple sub-states of an enzyme's active site (enzyme floppiness) and/or its substrate. Floppiness relates to the inherent flexibility of proteins, but also to conflicting demands, or trade-offs, between rate acceleration (the rate-determining chemical step) and catalytic turnover, or between turnover rate and accuracy. The study of futile encounters and active-site floppiness may contribute to a better understanding of enzyme catalysis, enzyme evolution, and improved enzyme design.

  15. Biochemical studies on changes associated with enzymes of ...

    African Journals Online (AJOL)

    SERVER

    2007-08-20

    Aug 20, 2007 ... enzymes of glucose metabolism in white spot syndrome virus ... links between stress caused by environmental changes and diseases and a ..... Webb (1973) also reported that stress affected qualitative and quantitative nature ...

  16. Histological and histochemical studies on the innervation of the respiratory organs of Calotes versicolor Daud. (Agamidae, Lacertilia).

    Science.gov (United States)

    Mathur, R; Kumar, S; Sharma, M C

    1979-12-01

    A study on the innervation of the respiratory organs of Calotes versicolor has been made using cholinesterase and silver reduction techniques. The tracheal and bronchial plexuses have been described. A number of nerve terminations along with synapses are reported. ChE activity has been observed on the periphery of the alveoli which is believed to be of importance in controlling the acetylcholine level, acting as a general tissue enzyme to prevent excessive accumulation of acetylcholine. Vascular nerve supply has been found to be largely of vagal origin.

  17. Enhanced Oil Recovery with Application of Enzymes

    DEFF Research Database (Denmark)

    Khusainova, Alsu

    behaviour of enzymes/proteins on the reservoir rocks was studied by application of the static adhesion tests and adsorption experiments on powders, as well as of dynamic flow-through experiments. It was established that enzymes are indeed significantly lost during the transport in the porous media due...... to detach oil from the calcite surface and was identified as the most promising group for further investigations. Wettability improvement due to protein adsorption on to the mineral was proposed as the main mechanism for EEOR. It was also proved that the enzyme molecules themselves caused change...... of the wetting state of calcite, while presence of stabilising ingredients did not interfere the results. Implementation of such a mechanism of enzymatic action under reservoir conditions might be limited by retention of the protein molecules in the porous medium. In order to verify this hypothesis, adsorption...

  18. Halophilic adaptation of enzymes.

    Science.gov (United States)

    Madern, D; Ebel, C; Zaccai, G

    2000-04-01

    It is now clear that the understanding of halophilic adaptation at a molecular level requires a strategy of complementary experiments, combining molecular biology, biochemistry, and cellular approaches with physical chemistry and thermodynamics. In this review, after a discussion of the definition and composition of halophilic enzymes, the effects of salt on their activity, solubility, and stability are reviewed. We then describe how thermodynamic observations, such as parameters pertaining to solvent-protein interactions or enzyme-unfolding kinetics, depend strongly on solvent composition and reveal the important role played by water and ion binding to halophilic proteins. The three high-resolution crystal structures now available for halophilic proteins are analyzed in terms of haloadaptation, and finally cellular response to salt stress is discussed briefly.

  19. Histochemical characterization, distribution and morphometric analysis of NADPH diaphorase neurons in the spinal cord of the agouti

    Directory of Open Access Journals (Sweden)

    Marco Aurelio M Freire

    2008-05-01

    Full Text Available We evaluated the neuropil distribution of the enzymes NADPH diaphorase (NADPH-d and cytochrome oxidase (CO in the spinal cord of the agouti, a medium-sized diurnal rodent, together with the distribution pattern and morphometrical characteristics of NADPH-d reactive neurons across different spinal segments. Neuropil labeling pattern was remarkably similar for both enzymes in coronal sections: reactivity was higher in regions involved with pain processing. We found two distinct types of NADPH-d reactive neurons in the agouti’s spinal cord: type I neurons had large, heavily stained cell bodies while type II neurons displayed relatively small and poorly stained somata. We concentrated our analysis on type I neurons. These were found mainly in the dorsal horn and around the central canal of every spinal segment, with a few scattered neurons located in the ventral horn of both cervical and lumbar regions. Overall, type I neurons were more numerous in the cervical region. Type I neurons were also found in the white matter, particularly in the ventral funiculum. Morphometrical analysis revealed that type I neurons located in the cervical region have dendritic trees that are more complex than those located in both lumbar and thoracic regions. In addition, NADPH-d cells located in the ventral horn had a larger cell body, especially in lumbar segments. The resulting pattern of cell body and neuropil distribution is in accordance with proposed schemes of segregation of function in the mammalian spinal cord.

  20. Immobilized enzymes in organic synthesis.

    Science.gov (United States)

    Mosbach, K

    1985-01-01

    The immobilization of enzymes and cells by different methods and the possible stabilization of immobilized preparations are discussed. An outlook on 'second generation enzyme technology', which involves immobilized multi-enzyme systems and coenzymes, is given with examples: the immobilization of dehydrogenases with their active sites facing one another, and systems containing NAD(H) coenzymes immobilized by coupling to dextran (in an enzyme electrode), to polyethylene glycol (in a membrane reactor), or to enzymes themselves. The use of immobilized enzymes to synthesize peptides and disaccharides is described.

  1. X-linked Angelman-like syndrome caused by Slc9a6 knockout in mice exhibits evidence of endosomal–lysosomal dysfunction

    Science.gov (United States)

    Strømme, Petter; Dobrenis, Kostantin; Sillitoe, Roy V.; Gulinello, Maria; Ali, Nafeeza F.; Davidson, Cristin; Micsenyi, Matthew C.; Stephney, Gloria; Ellevog, Linda; Klungland, Arne

    2011-01-01

    Mutations in solute carrier family 9 isoform 6 on chromosome Xq26.3 encoding sodium–hydrogen exchanger 6, a protein mainly expressed in early and recycling endosomes are known to cause a complex and slowly progressive degenerative human neurological disease. Three resulting phenotypes have so far been reported: an X-linked Angelman syndrome-like condition, Christianson syndrome and corticobasal degeneration with tau deposition, with each characterized by severe intellectual disability, epilepsy, autistic behaviour and ataxia. Hypothesizing that a sodium–hydrogen exchanger 6 deficiency would most likely disrupt the endosomal–lysosomal system of neurons, we examined Slc9a6 knockout mice with tissue staining and related techniques commonly used to study lysosomal storage disorders. As a result, we found that sodium–hydrogen exchanger 6 depletion leads to abnormal accumulation of GM2 ganglioside and unesterified cholesterol within late endosomes and lysosomes of neurons in selective brain regions, most notably the basolateral nuclei of the amygdala, the CA3 and CA4 regions and dentate gyrus of the hippocampus and some areas of cerebral cortex. In these select neuronal populations, histochemical staining for β-hexosaminidase activity, a lysosomal enzyme involved in the degradation of GM2 ganglioside, was undetectable. Neuroaxonal dystrophy similar to that observed in lysosomal disease was observed in the cerebellum and was accompanied by a marked and progressive loss of Purkinje cells, particularly in those lacking the expression of Zebrin II. On behavioural testing, Slc9a6 knockout mice displayed a discrete clinical phenotype attributable to motor hyperactivity and cerebellar dysfunction. Importantly, these findings show that sodium–hydrogen exchanger 6 loss of function in the Slc9a6-targeted mouse model leads to compromise of endosomal–lysosomal function similar to lysosomal disease and to conspicuous neuronal abnormalities in specific brain regions

  2. X-linked Angelman-like syndrome caused by Slc9a6 knockout in mice exhibits evidence of endosomal-lysosomal dysfunction.

    Science.gov (United States)

    Strømme, Petter; Dobrenis, Kostantin; Sillitoe, Roy V; Gulinello, Maria; Ali, Nafeeza F; Davidson, Cristin; Micsenyi, Matthew C; Stephney, Gloria; Ellevog, Linda; Klungland, Arne; Walkley, Steven U

    2011-11-01

    Mutations in solute carrier family 9 isoform 6 on chromosome Xq26.3 encoding sodium-hydrogen exchanger 6, a protein mainly expressed in early and recycling endosomes are known to cause a complex and slowly progressive degenerative human neurological disease. Three resulting phenotypes have so far been reported: an X-linked Angelman syndrome-like condition, Christianson syndrome and corticobasal degeneration with tau deposition, with each characterized by severe intellectual disability, epilepsy, autistic behaviour and ataxia. Hypothesizing that a sodium-hydrogen exchanger 6 deficiency would most likely disrupt the endosomal-lysosomal system of neurons, we examined Slc9a6 knockout mice with tissue staining and related techniques commonly used to study lysosomal storage disorders. As a result, we found that sodium-hydrogen exchanger 6 depletion leads to abnormal accumulation of GM2 ganglioside and unesterified cholesterol within late endosomes and lysosomes of neurons in selective brain regions, most notably the basolateral nuclei of the amygdala, the CA3 and CA4 regions and dentate gyrus of the hippocampus and some areas of cerebral cortex. In these select neuronal populations, histochemical staining for β-hexosaminidase activity, a lysosomal enzyme involved in the degradation of GM2 ganglioside, was undetectable. Neuroaxonal dystrophy similar to that observed in lysosomal disease was observed in the cerebellum and was accompanied by a marked and progressive loss of Purkinje cells, particularly in those lacking the expression of Zebrin II. On behavioural testing, Slc9a6 knockout mice displayed a discrete clinical phenotype attributable to motor hyperactivity and cerebellar dysfunction. Importantly, these findings show that sodium-hydrogen exchanger 6 loss of function in the Slc9a6-targeted mouse model leads to compromise of endosomal-lysosomal function similar to lysosomal disease and to conspicuous neuronal abnormalities in specific brain regions, which in concert

  3. Treating Wastewater With Immobilized Enzymes

    Science.gov (United States)

    Jolly, Clifford D.

    1991-01-01

    Experiments show enzymes are immobilized on supporting materials to make biocatalyst beds for treatment of wastewater. With suitable combination of enzymes, concentrations of various inorganic and organic contaminants, including ammonia and urea, reduced significantly.

  4. 栉孔扇贝(Chlamys (Azumapecten) farreri) 消化盲囊的组织学和组织化学的研究%A Histological and Histochemical Study on Digestive Diverticula of Farreris's Scallop, Chlamys (Azumapecten) farreri

    Institute of Scientific and Technical Information of China (English)

    绳秀珍; 刘晓云; 任素莲; 刘竹伞; 俞开康

    2001-01-01

    The structural and functional aspects of the digestive diverticula of Chlamys farreri were studied using light and transmission electron microscopy and histochemical methods. The epithelium of the digestive diverticula contains two types of cells, namely the digestive (absorptive) and secretory cells. The digestive cell is lined with microvilli, the presence of acid phosphatase activity in digestive diverticula suggest that these cells are actively involved in the absorption and intracellular digestion of food material, and the absorption role is supported by the presence of alkaline phosphatase in the digestive diverticula. Esterase and lipase are detected in the digestive cell. Lipid and glycogen are abundant throughout the digestive cell, showing that these cells are the sites of energy stores. The secretory cells contain abundant protein and RNA suggesting that they have synthetic function, together with their cytological feathers: numerous rough endoplasmic reticulum, Golgi complex and secretory-like granules support the view that in Farreri's scallop Chlamys (Azumapecten) farreri the secretory cell plays an important role in the production and release of digestive enzyme. The duct of the digestive diverticula is lined with cilia and microvilli, acid and alkaline phosphatase as well as lipase activity are detected in the duct cells. Therefore, it can produce digestive enzyme. Fe is present in the distal parts and the fragmentation sphere of the digestive cell, and no calcium exists in the digestive diverticula.%运用石蜡切片法、透射电镜技术及组织化学方法,对栉孔扇贝的消化盲囊进行了研究。结果表明:消化盲囊的腺上皮由消化(吸收)细胞和分泌细胞组成,消化(吸收)细胞顶端有微绒毛,细胞内有酸性磷酸酶、碱性磷酸酶、脂酶和酯酶活性,还含有糖原和脂肪。是进行食物的细胞内消化和营养物质吸收的主要场所,并有储存能量的功能;分泌细胞

  5. Kinetic Measurements for Enzyme Immobilization.

    Science.gov (United States)

    Cooney, Michael J

    2017-01-01

    Enzyme kinetics is the study of the chemical reactions that are catalyzed by enzymes, with a focus on their reaction rates. The study of an enzyme's kinetics considers the various stages of activity, reveals the catalytic mechanism of this enzyme, correlates its value to assay conditions, and describes how a drug or a poison might inhibit the enzyme. Victor Henri initially reported that enzyme reactions were initiated by a bond between the enzyme and the substrate. By 1910, Michaelis and Menten were advancing their work by studying the kinetics of an enzyme saccharase which catalyzes the hydrolysis of sucrose into glucose and fructose. They published their analysis and ever since the Michaelis-Menten equation has been used as the standard to describe the kinetics of many enzymes. Unfortunately, soluble enzymes must generally be immobilized to be reused for long times in industrial reactors. In addition, other critical enzyme properties have to be improved like stability, activity, inhibition by reaction products, and selectivity towards nonnatural substrates. Immobilization is by far the chosen process to achieve these goals.Although the Michaelis-Menten approach has been regularly adapted to the analysis of immobilized enzyme activity, its applicability to the immobilized state is limited by the barriers the immobilization matrix places upon the measurement of compounds that are used to model enzyme kinetics. That being said, the estimated value of the Michaelis-Menten coefficients (e.g., V max, K M) can be used to evaluate effects of immobilization on enzyme activity in the immobilized state when applied in a controlled manner. In this review enzyme activity and kinetics are discussed in the context of the immobilized state, and a few novel protocols are presented that address some of the unique constraints imposed by the immobilization barrier.

  6. Anatomia e histoquímica dos órgãos vegetativos de Polygonum hydropiperoides Michx., Polygonaceae Anatomical and histochemical characters of Polygonum hydropiperoides Michaux, Polygonaceae

    Directory of Open Access Journals (Sweden)

    Daniela E. Silva Lopes

    2009-09-01

    Full Text Available Polygonum hydropiperoides Michaux é uma espécie conhecida popularmente como "erva-de-bicho", amplamente utilizada na medicina tradicional como anti-hemorroidal, antiinflamatória e antidiarréica. O presente trabalho tem como objetivo a caracterização anatômica e histoquímica da folha, caule e raiz, que constituem a droga vegetal, visando estabelecer parâmetros para o controle de qualidade. O material vegetal foi fixado e submetido às técnicas usuais de microscopia de luz e aos testes histoquímicos. A folha é anfiestomática, dorsiventral e com estômatos paracíticos e anisocíticos. É comum a presença de estruturas secretoras como: tricomas glandulares capitados e glândulas epidérmicas e subepidérmicas em ambas as faces da lâmina foliar e também no caule. O material secretado pelas glândulas apresenta composição heterogênea de lipídios e flavonóides, segundo análises histoquímicas. Cristais de oxalato de cálcio e grãos de amido são freqüentes em células parenquimáticas da folha e do caule. Compostos fenólicos estão presentes na folha (parênquima paliçádico, no caule (parênquima cortical e floema e na raiz (parênquima cortical.P. hydropiperoides is popularly known as "erva-de-bicho", and it is used in traditional medicine as anti-hemorroidal, anti-inflammatory and antidiarrhoeic. The present work has as objective the anatomical and histochemical characterization of leaf, stem and root that are used as drug in order to offer elements for quality control. The botanical material was prepared for the usual optical and histochemical microtechniques. The leaf is amphistomatic, dorsiventral and paracytic and anisocytic stomata. It is common to find the presence of secretory structures as: capitate glandular trichomes and secretory glands on both epidermis, that are also present in stem. The secretion material is heterogeneous composition of lipids and flavonoids. Druses of calcium oxalate and starch are common in

  7. Histologia e histoquímica do magno, um dos segmentos do oviduto de Numida meleagris (Linné (Numididae, Galliformes Histological and histochemical of the magnum, a segment of oviduct of Numida meleagris (Linné (Numididae, Galliformes

    Directory of Open Access Journals (Sweden)

    Maria das Graças Ribeiro

    1997-01-01

    Full Text Available Numida meleagris (Linné, 1758 is an african bird that was brought to Brazil. As the bird adapted to the climate of this new habitat and spread ali around the country, it is nowadays part of our avifauna. The present study continues topographic and morphofunctional researches on the female genital apparatus of this species, since it describes histological and histochemical aspects of magnum, a segment of oviduct. Magnum was dissected and processed according to routine and histochemical staining procedures to detect glycogen and mucous substances in the epithelial tissue and mucous glands. Besides focusing morphological aspects, the study compares the data obtained with those of other species aiming to contribute to the enlargement of the knowledge on reproductive biology of brazilian birds what may be important to make their biological control easier.

  8. Odontogenic keratocysts: a clinical and histological study with special reference to enzyme histochemistry.

    Science.gov (United States)

    Magnusson, B C

    1978-02-01

    Of a total of 1,420 odontogenic cysts, 52 (3.3%) were diagnosed as odontogenic keratocysts. Clinical and histological findings in these 52 cysts are reported. Frozen sections of 26 of the keratocysts were incubated to show the following enzyme activities: NADH2- and NADPH2-diaphorase, glucose-6-phosphate dehydrogenase, glutamate dehydrogenase, acid phosphatase, leucine aminopeptidase and ATPase. Furthermore, keratinization was studied with the rhodamine B method and lipids with the oil red O, the OTAN and the acid hematein methods. Sections from epidermis, oral mucosa, radicular cysts, residual cysts and follicular cysts served as reference material. The oxidative enzymes showed strong activity in the keratocyst epithelium which contrasted with weak activity in the reference cysts. Acid phosphatase activity was weak in all epithelia except that in keratocysts, which displayed a marked activity. In the fibrous capsule of the keratocyst a high activity of leucine aminopeptidase was recorded. This high activity contrasted with a weak activity in the reference material. The significance of the histochemical results in relation to the aggressive behavior of the keratocyst is discussed.

  9. In situ histochemical localisation of alkaloids and acetogenins in the endosperm and embryonic axis of Annona macroprophyllata Donn. Sm. seeds during germination.

    Science.gov (United States)

    Brechú-Franco, A E; Laguna-Hernández, G; De la Cruz-Chacón, I; González-Esquinca, A R

    2016-02-01

    Currently, the Annonaceae family is characterised by the production of acetogenins (ACGs), and also by the biosynthesis of alkaloids, primarily benzylisoquinolines derived from tyrosine. The objective of this study was to confirm the presence of alkaloids and acetogenins in the idioblasts of the endosperm and the embryonic axis of A. macroprophyllata seeds in germination. The Dragendorff, Dittmar, Ellram, and Lugol reagents were used to test for alkaloids, and Kedde's reagent was used to determine the presence of acetogenins in fresh sections of the endosperm and embryonic axis of seeds after twelve days of germination. A positive reaction was observed for all the reagents, and the presence of alkaloids and acetogenins was confirmed in the idioblasts of the endosperm and those involved in the differentiation of the embryonic axis of the developing seedling. We concluded that the idioblasts store both metabolites, acetogenins and alkaloids. Beginning at differentiation, the idioblasts of the embryonic axis simultaneously biosynthesise acetogenins and alkaloids that are characteristic of the species during the development of the seedling. The method used here can be applied to histochemically confirm the presence of acetogenins and alkaloids in tissues and structures of the plant in different stages of its life cycle.

  10. [Changes in the innervation of the uterus during pregnancy and following parturition. Histochemical and electron microscopic observations in rat and humans].

    Science.gov (United States)

    Yamada, M

    1988-02-01

    A histochemical, electronmicroscopic study was made of time changes in uterine nerves during and after pregnancy in rats. Uterine observations in pregnant humans were also conducted with the following results. 1. Except at the oviductal end, the catecholamine (CA) fluorescence of adrenergic nerves distributed in the uterine horn myometrium begins to decrease around the implantation sites, and this spreads peripherally until most of the fluorescence in the uterine horns, including the mesometrium, disappears directly after parturition. 2. In lactating rats, fluorescence is almost completely restored by 1 month after parturition. But in non-lactating rats, recovery of fluorescence after parturition is delayed about 3 months. 3. The activity of acetylcholinesterase (AChE) in rat uterine cholinergic nerves clearly decreased during pregnancy. 4. In the early, middle and late periods of pregnancy, axonal swelling, agglutination of axonal cytoplasm and mitochondrial breakdown were observed, but no marked degeneration appeared in virgin rat uteri. 5. In humans, there was a sporadic distribution of adrenergic nerves in the uterine body myometrium, but fluorescent fibers were present in the terminal period of pregnancy. These results suggest that rat uterine nerves, especially most adrenergic nerves, degenerate and regenerate throughout pregnancy, parturition and puerperium. Apparently a localized humoral factor from the placenta and systemic endocrine environment affected by lactation or non-lactation is involved in the process.

  11. In situ histochemical localisation of alkaloids and acetogenins in the endosperm and embryonic axis of Annona macroprophyllata Donn. Sm. seeds during germination

    Directory of Open Access Journals (Sweden)

    A.E. Brechú-Franco

    2016-02-01

    Full Text Available Currently, the Annonaceae family is characterised by the production of acetogenins (ACGs, and also by the biosynthesis of alkaloids, primarily benzylisoquinolines derived from tyrosine. The objective of this study was to confirm the presence of alkaloids and acetogenins in the idioblasts of the endosperm and the embryonic axis of A. macroprophyllata seeds in germination. The Dragendorff, Dittmar, Ellram, and Lugol reagents were used to test for alkaloids, and Kedde’s reagent was used to determine the presence of acetogenins in fresh sections of the endosperm and embryonic axis of seeds after twelve days of germination. A positive reaction was observed for all the reagents, and the presence of alkaloids and acetogenins was confirmed in the idioblasts of the endosperm and those involved in the differentiation of the embryonic axis of the developing seedling. We concluded that the idioblasts store both metabolites, acetogenins and alkaloids. Beginning at differentiation, the idioblasts of the embryonic axis simultaneously biosynthesise acetogenins and alkaloids that are characteristic of the species during the development of the seedling. The method used here can be applied to histochemically confirm the presence of acetogenins and alkaloids in tissues and structures of the plant in different stages of its life cycle.

  12. Anatomical and histological structure of the tongue and histochemical characteristics of the lingual salivary glands in the Chukar partridge (Alectoris chukar, Gray 1830).

    Science.gov (United States)

    Erdoğan, S; Sağsöz, H; Akbalik, M E

    2012-01-01

    1. The aim of the study was to examine the morphology of the tongue and the histochemical features of the lingual salivary glands in this species. 2. The tongue was elongated, terminating in a rather sharp, dagger-like apex. On the surface of the tongue and situated between the body and root of the tongue, two rows of conical papillae, the sharp apices of which pointed towards the posterior part of the tongue, were observed. The keratinised epithelium lining the dorsal surface lacked typical gustatory papillae. However, it was observed that taste buds were present in the epithelium of the lingual body and root. The tongue was supported by a structure composed of hyaline cartilage, the paraglossum, which extended from the lingual root to the apex. Simple branched tubular glands, which were encapsulated by connective tissue, were embedded within the submucosa in the body (anterior salivary glands) and root (posterior salivary glands) of the tongue. It was observed that the secretion of the lingual glands contained neutral mucins, proteoglycans containing carboxylic acid, weak and strong sulphated groups, N-acetylated sialomucins, but lacked glycogen. 3. It was demonstrated that, the general morphological features, papillary distribution of the tongue and the histological structure of the mucosa epithelium and the supportive elements displayed similarity to those of other domestic avian species. It was also determined that, in view of the particular feeding types, in the partridge, the presence of the papillary crest was not correlated with diet.

  13. Measuring the Enzyme Activity of Arabidopsis Deubiquitylating Enzymes.

    Science.gov (United States)

    Kalinowska, Kamila; Nagel, Marie-Kristin; Isono, Erika

    2016-01-01

    Deubiquitylating enzymes, or DUBs, are important regulators of ubiquitin homeostasis and substrate stability, though the molecular mechanisms of most of the DUBs in plants are not yet understood. As different ubiquitin chain types are implicated in different biological pathways, it is important to analyze the enzyme characteristic for studying a DUB. Quantitative analysis of DUB activity is also important to determine enzyme kinetics and the influence of DUB binding proteins on the enzyme activity. Here, we show methods to analyze DUB activity using immunodetection, Coomassie Brilliant Blue staining, and fluorescence measurement that can be useful for understanding the basic characteristic of DUBs.

  14. Sertraline-induced pseudocholinesterase enzyme deficiency

    Directory of Open Access Journals (Sweden)

    Beyazit Zencirci

    2010-11-01

    Full Text Available Beyazit ZencirciMOSTAS Private Health Hospital, Department of Anesthesiology, Kahramanmaras, TurkeyAbstract: A 47-year-old Turkish male was scheduled for laparoscopic cholecystectomy under general anesthesia. The patient had 2 operations 28 and 19 years ago under general anesthesia. It was learned that the patient was administered succinylcholine during both of these previous operations and that he did not have a history of prolonged recovery or postoperative apnea. The patient had been using sertraline for 3 years before the operation. Pseudocholinesterase is a drug-metabolizing enzyme responsible for hydrolysis of the muscle-relaxant drugs mivacurium and succinylcholine. Deficiency of this enzyme from any cause can lead to prolonged apnea and paralysis following administration of mivacurium and succinylcholine. The diagnosis of pseudocholinesterase enzyme deficiency can be made after careful clinic supervision and peripheral nerve stimulator monitoring. A decrease in the activity of pseudocholinesterase enzyme and a decline in the block effect over time will help verify the diagnosis. Our patient’s plasma cholinesterase was found to have low activity. Instead of pharmacological interventions that may further complicate the situation in such cases, the preferred course of action should be to wait until the block effect declines with the help of sedation and mechanical ventilation. In our case, the prolonged block deteriorated in the course of time before any complications developed.Keywords: mivacurium, pseudocholinesterase deficiency, sertraline

  15. Histomorphological and Histochemical Alterations Following Short-term Inhalation Exposure to Sulfur Mustard on Visceral Organs of Mice

    Institute of Scientific and Technical Information of China (English)

    S. C. PANT; R. VIJAYARAGHAVAN

    1999-01-01

    Toxic effects of inhaled sulfur mustard (SM) on the histology of visceral organs was investigated by exposing mice to 84.6mg/m3 for 1 h duration, using controlled single exposure conditions. A progressive fall in body weight from third day onwards was noticed. Light microscopic examination of the pulmonary tissue of these animals at 6 h post exposure revealed that the tracheobronchial epithelium remained intact, but was infiltrated by inflammatorv cells. By 24 h post exposure, the mucosecretory cells were destroyed. The inflammatory reaction was maximum at 48 h. Bv 7th day post exposure there was swelling and vacuolation of lung parenchymal cells and thrombi formation. In addition SM caused congestion and hemorrhage at alveolar level. SM also caused granulovacuolar degeneration with perinuclear clumping of the cytoplasm of hepatocytes and renal parenchymal cells. Renal lesions were characterized by congestion and hemorrhage. Among visceral tissues, maximum atrophy was observed in spleen. Distribution of lesions increased with post exposure period. The maximum lesions were observed at 7th day post-exposure.

  16. Enzyme molecules in solitary confinement.

    Science.gov (United States)

    Liebherr, Raphaela B; Gorris, Hans H

    2014-09-12

    Large arrays of homogeneous microwells each defining a femtoliter volume are a versatile platform for monitoring the substrate turnover of many individual enzyme molecules in parallel. The high degree of parallelization enables the analysis of a statistically representative enzyme population. Enclosing individual enzyme molecules in microwells does not require any surface immobilization step and enables the kinetic investigation of enzymes free in solution. This review describes various microwell array formats and explores their applications for the detection and investigation of single enzyme molecules. The development of new fabrication techniques and sensitive detection methods drives the field of single molecule enzymology. Here, we introduce recent progress in single enzyme molecule analysis in microwell arrays and discuss the challenges and opportunities.

  17. Enzyme Molecules in Solitary Confinement

    Directory of Open Access Journals (Sweden)

    Raphaela B. Liebherr

    2014-09-01

    Full Text Available Large arrays of homogeneous microwells each defining a femtoliter volume are a versatile platform for monitoring the substrate turnover of many individual enzyme molecules in parallel. The high degree of parallelization enables the analysis of a statistically representative enzyme population. Enclosing individual enzyme molecules in microwells does not require any surface immobilization step and enables the kinetic investigation of enzymes free in solution. This review describes various microwell array formats and explores their applications for the detection and investigation of single enzyme molecules. The development of new fabrication techniques and sensitive detection methods drives the field of single molecule enzymology. Here, we introduce recent progress in single enzyme molecule analysis in microwell arrays and discuss the challenges and opportunities.

  18. A Simple Structure Model for Enzyme Production by Phanerochaete chrysosporium

    Institute of Scientific and Technical Information of China (English)

    岑沛霖; 郑重鸣; FOOYinDin; JefferyPhilipObbard; 林建平

    2003-01-01

    In order to understand the behavior of ligninolytic enzyme production by white rot fungi Phanerochaete chrysosporium, study on time courses and a mathematical model for the production of lignin peroxidase (LiP) and manganese peroxidase (MnP) of the fungi was undertaken. Based on the Monod-Jacob operon model, the ligninolytic enzyme would be synthesized in the absence of a related repressor. The repressor is assumed to be active in the presence of ammonia nitrogen, and as combined as co-repressor, it causes the inhibition of enzyme synthesis. The model can explain the mechanism of extracellular ligninolytic enzyme production by white rot fungi. The results,as predicted by the model, correspond closely to those observed in experimental studies. In addition, some light is also shed on unmeasured variables, such as the concentrations of repressor and mRNA that are related to the enzyme synthesis.

  19. Antioxidant enzyme levels in cancer

    OpenAIRE

    Oberley, T. D.; Oberley, L W

    1997-01-01

    Normal cells are protected by antioxidant enzymes from the toxic effects of high concentrations of reactive oxygen species generated during cellular metabolism. Even though cancer cells generate reactive oxygen species, it has been demonstrated biochemically that antioxidant enzyme levels are low in most animal and human cancers. However, a few cancer types have been found to have elevated levels of antioxidant enzymes, particularly manganese superoxide dismuta...

  20. Heat Stable Enzymes from Thermophiles

    Science.gov (United States)

    1998-02-01

    ultrafiltration and microfiltration that might be suitable. These utilize hollow fiber membranes manufactured in such a manner that they are free of...words) Alkaline phosphatase is widely used in the military and civilian sectors . Commercially available enzyme from calf intestine is the weak link in...widely used enzymes with numerous uses in both the military and civilian sectors . The commercially available enzyme from calf intestine breaks down

  1. Multi-enzyme Process Modeling

    DEFF Research Database (Denmark)

    Andrade Santacoloma, Paloma de Gracia

    The subject of this thesis is to develop a methodological framework that can systematically guide mathematical model building for better understanding of multi-enzyme processes. In this way, opportunities for process improvements can be identified by analyzing simulations of either existing...... are affected (in a positive or negative way) by the presence of the other enzymes and compounds in the media. In this thesis the concept of multi-enzyme in-pot term is adopted for processes that are carried out by the combination of enzymes in a single reactor and implemented at pilot or industrial scale...

  2. Enzyme therapeutics for systemic detoxification.

    Science.gov (United States)

    Liu, Yang; Li, Jie; Lu, Yunfeng

    2015-08-01

    Life relies on numerous biochemical processes working synergistically and correctly. Certain substances disrupt these processes, inducing living organism into an abnormal state termed intoxication. Managing intoxication usually requires interventions, which is referred as detoxification. Decades of development on detoxification reveals the potential of enzymes as ideal therapeutics and antidotes, because their high substrate specificity and catalytic efficiency are essential for clearing intoxicating substances without adverse effects. However, intrinsic shortcomings of enzymes including low stability and high immunogenicity are major hurdles, which could be overcome by delivering enzymes with specially designed nanocarriers. Extensive investigations on protein delivery indicate three types of enzyme-nanocarrier architectures that show more promise than others for systemic detoxification, including liposome-wrapped enzymes, polymer-enzyme conjugates, and polymer-encapsulated enzymes. This review highlights recent advances in these nano-architectures and discusses their applications in systemic detoxifications. Therapeutic potential of various enzymes as well as associated challenges in achieving effective delivery of therapeutic enzymes will also be discussed.

  3. Digestive Enzyme Replacement Therapy: Pancreatic Enzymes and Lactase.

    Science.gov (United States)

    Felicilda-Reynaldo, Rhea Faye D; Kenneally, Maria

    2016-01-01

    Maldigestion occurs when digestive enzymes are lacking to help break complex food components into absorbable nutrients within the gastrointestinal tract. Education is needed to help patients manage the intricacies of digestive enzyme replacement therapies and ensure their effectiveness in reducing symptoms of maldigestion.

  4. Enzymic synthesis of isoflavones.

    Science.gov (United States)

    Kochs, G; Grisebach, H

    1986-03-03

    The NADPH and oxygen-dependent conversion of (2S)-naringenin to genistein catalyzed by a microsomal preparation from elicitor-treated soybean cell suspension cultures has been resolved into two steps. In the first step (2S)-naringenin is converted to a product (P-2) which yields genistein in a second step. The chemical behaviour of P-2 and its ultraviolet and mass spectral data are consistent with a 2-hydroxyisoflavanone structure. The conversion of (2S)-naringenin to P-2 requires NADPH, oxygen and cytochrome P-450. The participation of cytochrome P-450 was demonstrated by CO inhibition of the reaction and its partial reversal by light, and by inhibition with typical cytochrome P-450 inhibitors. On a Percoll gradient the membrane fraction which catalyzes P-2 formation coincides with marker enzymes for the endoplasmic reticulum and with the position of cytochrome P-450. Enzymatic activity for conversion of P-2 to genistein is mainly present in the supernatant of the 160 000 X g fraction. This reaction, formally a dehydration, does not require NADPH or oxygen.

  5. Deubiquitylating enzymes and disease

    Directory of Open Access Journals (Sweden)

    Baker Rohan T

    2008-10-01

    Full Text Available Abstract Deubiquitylating enzymes (DUBs can hydrolyze a peptide, amide, ester or thiolester bond at the C-terminus of UBIQ (ubiquitin, including the post-translationally formed branched peptide bonds in mono- or multi-ubiquitylated conjugates. DUBs thus have the potential to regulate any UBIQ-mediated cellular process, the two best characterized being proteolysis and protein trafficking. Mammals contain some 80–90 DUBs in five different subfamilies, only a handful of which have been characterized with respect to the proteins that they interact with and deubiquitylate. Several other DUBs have been implicated in various disease processes in which they are changed by mutation, have altered expression levels, and/or form part of regulatory complexes. Specific examples of DUB involvement in various diseases are presented. While no specific drugs targeting DUBs have yet been described, sufficient functional and structural information has accumulated in some cases to allow their rapid development. Publication history Republished from Current BioData's Targeted Proteins database (TPdb; http://www.targetedproteinsdb.com.

  6. [The rise of enzyme engineering in China].

    Science.gov (United States)

    Li, Gaoxiang

    2015-06-01

    Enzyme engineering is an important part of the modern biotechnology. Industrial biocatalysis is considered the third wave of biotechnology following pharmaceutical and agricultural waves. In 25 years, China has made a mighty advances in enzyme engineering research. This review focuses on enzyme genomics, enzyme proteomics, biosynthesis, microbial conversion and biosensors in the Chinese enzyme engineering symposiums and advances in enzyme preparation industry in China.

  7. Computational enzyme design: transitioning from catalytic proteins to enzymes.

    Science.gov (United States)

    Mak, Wai Shun; Siegel, Justin B

    2014-08-01

    The widespread interest in enzymes stem from their ability to catalyze chemical reactions under mild and ecologically friendly conditions with unparalleled catalytic proficiencies. While thousands of naturally occurring enzymes have been identified and characterized, there are still numerous important applications for which there are no biological catalysts capable of performing the desired chemical transformation. In order to engineer enzymes for which there is no natural starting point, efforts using a combination of quantum chemistry and force-field based protein molecular modeling have led to the design of novel proteins capable of catalyzing chemical reactions not catalyzed by naturally occurring enzymes. Here we discuss the current status and potential avenues to pursue as the field of computational enzyme design moves forward.

  8. Stability of Enzymes in Granular Enzyme Products for Laundry Detergents

    DEFF Research Database (Denmark)

    Biran, Suzan; Bach, Poul; Simonsen, Ole

    . However, incorporating enzymes in detergent formulations gives rise to numerous practical problems due to their incompatibility with and stability against various detergent components. In powdered detergent formulations, these issues can be partly overcome by physically isolating the enzymes in separate...... of this study. The inactivation kinetics of technical grade enzyme powder was determined in a newly developed experimental setup, which was simple and effective and provided a better control over test conditions and fast sample generation. The method was based on the generation of hydrogen peroxide vapor...... the moisture is believed to play an important role in the stability of proteins, the monolayer hydration level of Savinase® was experimentally determined and theoretically calculated. Adsorbed moisture was found to have 3 a negative effect on enzyme activity. Below monolayer hydration level, the enzyme...

  9. Ecological effects of atmospheric nitrogen deposition on soil enzyme activity

    Institute of Scientific and Technical Information of China (English)

    WANG Cong-yan; Lv Yan-na; LIU Xue-yan Liu; WANG Lei

    2013-01-01

    The continuing increase in human activities is causing global changes such as increased deposition of atmospheric nitrogen.There is considerable interest in understanding the effects of increasing atmospheric nitrogen deposition on soil enzyme activities,specifically in terms of global nitrogen cycling and its potential future contribution to global climate change.This paper summarizes the ecological effects of atmospheric nitrogen deposition on soil enzyme activities,including size-effects,stage-effects,site-effects,and the effects of different levels and forms of atmospheric nitrogen deposition.We discuss needs for further research on the relationship between atmospheric nitrogen deposition and soil enzymes.

  10. Petroleum-degrading enzymes: bioremediation and new prospects.

    Science.gov (United States)

    Peixoto, R S; Vermelho, A B; Rosado, A S

    2011-01-01

    Anthropogenic forces, such as petroleum spills and the incomplete combustion of fossil fuels, have caused an accumulation of petroleum hydrocarbons in the environment. The accumulation of petroleum and its derivatives now constitutes an important environmental problem. Biocatalysis introduces new ways to improve the development of bioremediation strategies. The recent application of molecular tools to biocatalysis may improve bioprospecting research, enzyme yield recovery, and enzyme specificity, thus increasing cost-benefit ratios. Enzymatic remediation is a valuable alternative as it can be easier to work with than whole organisms, especially in extreme environments. Furthermore, the use of free enzymes avoids the release of exotic or genetically modified organisms (GMO) in the environment.

  11. Steroidogenic enzyme histochemistry in the testis of Sprague Dawley rats following the administration the water extracts from Carica papaya seed.

    Science.gov (United States)

    Uche-Nwachi, E O; Mitchell, C V; McEwen, C

    2011-01-01

    Water extracts from pawpaw seed have been reported to reversibly decrease the testicular weight and to suppress spermatogenesis, and fertility of Wistar rats. The reversible changes become evident, 30 - 45 days after the withdrawal of the extract. The possible effect of this extract on the activities of steroidogenic enzymes of the testis has not been investigated. Water extract of papaya seeds was administered to male Sprague Dawley rats ad libitum for 84 days. Following the discontinuation of the extracts, ten rats each were sacrificed on days 0, 10, 20 and 30 after the withdrawal. Their testes were quickly dissected out and frozen. Cryostat sections, 10µm thick were cut. These sections were used for immunohistochemical stains for side chain cleavage enzyme and aromatase, and for histochemical stains for 17-β Hydroxysteroid dehydrogenase, 3-β Hydroxysteroid dehydrogenase. We conclude that the water extract of papaya seed suppresses the activities of steroidogenic enzymes in the testis of Sprague Dawley rats, and that this may contribute to reversible suppression of spermatogenesis, a property that gives a possible male contraceptive potential.

  12. Lignocellulolytic enzyme production of Pleurotus ostreatus growth in agroindustrial wastes.

    Science.gov (United States)

    da Luz, José Maria Rodrigues; Nunes, Mateus Dias; Paes, Sirlaine Albino; Torres, Denise Pereira; de Cássia Soares da Silva, Marliane; Kasuya, Maria Catarina Megumi

    2012-10-01

    The mushroom Pleurotus ostreatus has nutritional and medicinal characteristics that depend on the growth substrate. In nature, this fungus grows on dead wood, but it can be artificially cultivated on agricultural wastes (coffee husks, eucalyptus sawdust, corncobs and sugar cane bagasse). The degradation of agricultural wastes involves some enzyme complexes made up of oxidative (laccase, manganese peroxidase and lignin peroxidase) and hydrolytic enzymes (cellulases, xylanases and tanases). Understanding how these enzymes work will help to improve the productivity of mushroom cultures and decrease the potential pollution that can be caused by inadequate discharge of the agroindustrial residues. The objective of this work was to assess the activity of the lignocellulolytic enzymes produced by two P. ostreatus strains (PLO 2 and PLO 6). These strains were used to inoculate samples of coffee husks, eucalyptus sawdust or eucalyptus bark add with or without 20 % rice bran. Every five days after substrate inoculation, the enzyme activity and soluble protein concentration were evaluated. The maximum activity of oxidative enzymes was observed at day 10 after inoculation, and the activity of the hydrolytic enzymes increased during the entire period of the experiment. The results show that substrate composition and colonization time influenced the activity of the lignocellulolytic enzymes.

  13. Lignocellulolytic enzyme production of Pleurotus ostreatus growth in agroindustrial wastes

    Directory of Open Access Journals (Sweden)

    José Maria Rodrigues da Luz

    2012-12-01

    Full Text Available The mushroom Pleurotus ostreatus has nutritional and medicinal characteristics that depend on the growth substrate. In nature, this fungus grows on dead wood, but it can be artificially cultivated on agricultural wastes (coffee husks, eucalyptus sawdust, corncobs and sugar cane bagasse. The degradation of agricultural wastes involves some enzyme complexes made up of oxidative (laccase, manganese peroxidase and lignin peroxidase and hydrolytic enzymes (cellulases, xylanases and tanases. Understanding how these enzymes work will help to improve the productivity of mushroom cultures and decrease the potential pollution that can be caused by inadequate discharge of the agroindustrial residues. The objective of this work was to assess the activity of the lignocellulolytic enzymes produced by two P. ostreatus strains (PLO 2 and PLO 6. These strains were used to inoculate samples of coffee husks, eucalyptus sawdust or eucalyptus bark add with or without 20 % rice bran. Every five days after substrate inoculation, the enzyme activity and soluble protein concentration were evaluated. The maximum activity of oxidative enzymes was observed at day 10 after inoculation, and the activity of the hydrolytic enzymes increased during the entire period of the experiment. The results show that substrate composition and colonization time influenced the activity of the lignocellulolytic enzymes.

  14. Enzymic hydrolysis of chlorella cells

    Energy Technology Data Exchange (ETDEWEB)

    Khraptsova, G.I.; Tsaplina, I.A.; Burdenko, L.G.; Khoreva, S.L.; Loginova, L.G.

    1981-01-01

    Treatment of C. ellipsoidea, C. pyrenoidosa, and C. vulgaris with cellulolytic enzymes (from Aspergillus terreus) and pectofoetidin p10x (from A. foetidus) resulted in the degradation and lysis of the algae cells. The cells were more sensitive to cellulase than to pectinase. The combination of both enzymes produced a synergistic effect on cell lysis.

  15. An enzyme with rhamnogalacturonase activity.

    NARCIS (Netherlands)

    Kovod, L.V.; Dalboge, H.; Andersen, L.N.; Kauppinen, M.; Christgan, S.; Heldt-Hansen, H.P.; Christophersen, C.; Nielsen, P.M.; Voragen, A.G.J.; Schols, H.A.

    1994-01-01

    An enzyme exhibiting rhamnogalacturonase activity, which enzyme: a) is encoded by the DNA sequence shown in SEQ ID No. 1 or a sequence homologous thereto encoding a polypeptide with RGase activity, b) has the amino acid sequence shown in SEQ ID No. 2 or an analogue thereof, c) is reactive with an

  16. Phage lytic enzymes: a history

    Institute of Scientific and Technical Information of China (English)

    David; Trudil

    2015-01-01

    There are many recent studies regarding the efficacy of bacteriophage-related lytic enzymes: the enzymes of ‘bacteria-eaters’ or viruses that infect bacteria. By degrading the cell wall of the targeted bacteria, these lytic enzymes have been shown to efficiently lyse Gram-positive bacteria without affecting normal flora and non-related bacteria. Recent studies have suggested approaches for lysing Gram-negative bacteria as well(Briersa Y, et al., 2014). These enzymes include: phage-lysozyme, endolysin, lysozyme, lysin, phage lysin, phage lytic enzymes, phageassociated enzymes, enzybiotics, muralysin, muramidase, virolysin and designations such as Ply, PAE and others. Bacteriophages are viruses that kill bacteria, do not contribute to antimicrobial resistance, are easy to develop, inexpensive to manufacture and safe for humans, animals and the environment. The current focus on lytic enzymes has been on their use as anti-infectives in humans and more recently in agricultural research models. The initial translational application of lytic enzymes, however, was not associated with treating or preventing a specifi c disease but rather as an extraction method to be incorporated in a rapid bacterial detection assay(Bernstein D, 1997).The current review traces the translational history of phage lytic enzymes–from their initial discovery in 1986 for the rapid detection of group A streptococcus in clinical specimens to evolving applications in the detection and prevention of disease in humans and in agriculture.

  17. A Novel Splice-Site Mutation in Angiotensin I-Converting Enzyme (ACE) Gene, c.3691+1G.A (IVS25+1G.A), Causes a Dramatic Increase in Circulating ACE through Deletion of the Transmembrane Anchor

    NARCIS (Netherlands)

    Persu, A.; Lambert, M.; Deinum, J.; Cossu, M.; Visscher, N. de; Irenge, L.; Ambroise, J.; Minon, J.M.; Nesterovitch, A.B.; Churbanov, A.; Popova, I.A.; Danilov, S.M.; Danser, A.H.; Gala, J.L.

    2013-01-01

    BACKGROUND: Angiotensin-converting enzyme (ACE) (EC 4.15.1) metabolizes many biologically active peptides and plays a key role in blood pressure regulation and vascular remodeling. Elevated ACE levels are associated with different cardiovascular and respiratory diseases. METHODS AND RESULTS: Two Bel

  18. Moonlighting enzymes in parasitic protozoa.

    Science.gov (United States)

    Collingridge, Peter W; Brown, Robert W B; Ginger, Michael L

    2010-08-01

    Enzymes moonlight in a non-enzymatic capacity in a diverse variety of cellular processes. The discovery of these non-enzymatic functions is generally unexpected, and moonlighting enzymes are known in both prokaryotes and eukaryotes. Importantly, this unexpected multi-functionality indicates that caution might be needed on some occasions in interpreting phenotypes that result from the deletion or gene-silencing of some enzymes, including some of the best known enzymes from classic intermediary metabolism. Here, we provide an overview of enzyme moonlighting in parasitic protists. Unequivocal and putative examples of moonlighting are discussed, together with the possibility that the unusual biological characteristics of some parasites either limit opportunities for moonlighting to arise or perhaps contribute to the evolution of novel proteins with clear metabolic ancestry.

  19. Statistical Mechanics of Allosteric Enzymes.

    Science.gov (United States)

    Einav, Tal; Mazutis, Linas; Phillips, Rob

    2016-07-07

    The concept of allostery in which macromolecules switch between two different conformations is a central theme in biological processes ranging from gene regulation to cell signaling to enzymology. Allosteric enzymes pervade metabolic processes, yet a simple and unified treatment of the effects of allostery in enzymes has been lacking. In this work, we take a step toward this goal by modeling allosteric enzymes and their interaction with two key molecular players-allosteric regulators and competitive inhibitors. We then apply this model to characterize existing data on enzyme activity, comment on how enzyme parameters (such as substrate binding affinity) can be experimentally tuned, and make novel predictions on how to control phenomena such as substrate inhibition.

  20. Effects of Melatonin on the Cerebellum of Infant Rat Following Kaolin-Induced Hydrocephalus: a Histochemical and Immunohistochemical Study.

    Science.gov (United States)

    Uyanıkgil, Yiğit; Turgut, Mehmet; Baka, Meral

    2017-02-01

    Hydrocephalus is a developmental disorder causing abnormally collected cerebrospinal fluid within the cerebral ventricles. It leads to bigger skulls and many dysfunctions related to the nervous system. Here, we addressed whether exogenous melatonin administration could reverse the clinical features of kaolin-induced hydrocephalus in infantile rats. A controlled double-blinded study was conducted in 2-week-old 45 Wistar albino rats, which were divided into three groups: Group A, the control group, received intracisternal sham injection with solely the needle insertion; group B, the hydrocephalus group, was treated with isotonic NaCl after kaolin injection; and group C, the hydrocephalus + melatonin group, was given i.p. exogenous melatonin at a dose of 0.5 mg/100 g body weight after kaolin injection. Histological and immunohistochemical analyses were performed after the induction of hydrocephalus and melatonin administration. Glial fibrillary acidic protein was stained by immunohistochemical method. TUNEL method was used to define and quantitate apoptosis in the cerebellar tissues. Statistical analysis was performed by nonparametric Kruskal-Wallis H test, and once significance was determined among means, post hoc pairwise comparisons were carried out using Mann-Whitney U test. We found that melatonin administration significantly ameliorated ratio of substantia grisea area/substantia alba area in the cerebellum of infantile rats. Histologically, there was a significant reduction in the number of cerebellar apoptotic cells after the hydrocephalus induced by kaolin (P kaolin-induced hydrocephalus. Nevertheless, further studies are needed to suggest melatonin as a candidate protective drug in children with hydrocephalus.

  1. Influence of Autologus Adipose Derived Stem Cells and PRP on Regeneration of Dehiscence-Type Defects in Alveolar Bone: A Comparative Histochemical and Histomorphometric Study in Dogs

    Science.gov (United States)

    Aziz Aly, Lobna Abdel; El- Menoufy, Hala; Hassan, Amal; Ragae, Alyaa; Atta, Hazem Mahmoud; Roshdy, Nagwa Kamal; Rashed, Laila Ahmed; Sabry, Dina

    2011-01-01

    Background and Objectives: Autogenous bone grafts is considered to be the best choice for reconstructive surgery. Adipose Derived Stromal Cells (ASCs) represents a promising tool for new clinical concepts in supporting cellular therapy. The goal of our study was to investigate bone regeneration following application of autologous ASCs with or without Platelet-Rich Plasma (PRP) at dehiscence-type defects in alveolar bone in dogs. Methods and Results: Standardized buccal dehiscence defects (4× 3×3 mm) were surgically created in eighteen dogs, the defects were grafted with either ASCs -PRP, ASCs alone, or without grafting material. Three months later; a bone core was harvested from grafted and non grafted sites for histological, histochemical and histomorphometric assessment. There was no evidence of inflammation or adverse tissue reaction with either treatment. Defects grafted with ASCs-PRP showed a significantly higher result (p≤ 0.05), with a mean area % of spongy bone and compact bone of (64.96±5.37 and 837.62±24.95), compared to ASCs alone (47.65±1.43 and 661.92±12.65) and without grafting (33.55± 1.74 and 290.85±7.27) respectively. The area % of lamellated bone increased significantly reaching its highest level in group A followed by group B. Also a significant increase in area % of neutral mucopolysaccharides and calcified reactivity of Masson|s Trichrome stain in groups A and B compared to group C was obtained. Conclusions: Our results suggest that, the addition of PRP to ASCs enhances bone formation after 3 months and may be clinically effective in accelerating postsurgical healing in both periodontal and maxillofacial surgical applications. PMID:24298335

  2. Histochemical examination of the effects of high-dose 1,25(OH)2D3 on bone remodeling in young growing rats.

    Science.gov (United States)

    Sun, Jing; Sun, Bao; Wang, Wei; Han, Xiuchun; Liu, Hongrui; Du, Juan; Feng, Wei; Liu, Bo; Amizuka, Norio; Li, Minqi

    2016-08-01

    Vitamin D has an anabolic effect on bone developmental processes and is involved in maintaining skeletal integrity. In recent years, pediatric cases of vitamin D intoxication have attracted attention. Therefore, the aim of this study was to investigate the influence of long-term administration of physiologically-high-dose calcitriol (1,25(OH)2D3) on bone remodeling in young developing rats. Neonatal rats received once-daily subcutaneous injection of calcitriol (250 ng/kg body weight), or PBS only as a control, for 3 weeks. At 1, 2 and 4 weeks' post-administration, rats were sacrificed and fixed by transcardial perfusion with 4 % paraformaldehyde, following which tibiae were extracted for histochemical analysis. Compared with the control group, the number of tartrate-resistant acid phosphatase- and Cathepsin K-positive osteoclasts were significantly increased, and the expression of alkaline phosphatase in osteoblasts was decreased in trabecular bone of rats administered high-dose 1,25(OH)2D3, leading to decreased trabecular bone volume. In addition, the expression of receptor activator of nuclear factor kappa-B ligand (RANKL) was increased, while that of osteoprotegerin was weaker in osteoblasts in the experimental group compared with the control group. Moreover, there was weaker immunoreactivity for EphrinB2 in osteoclasts and EphB4 in osteoblasts of trabecular bone in the experimental group compared with the control group. These findings suggest that long-term use of physiologically-high dose calcitriol may result in bone loss through RANKL/RANK/osteoprotegerin and EphrinB2-EphB4 signaling pathways, and that these negative effects could continue after drug withdrawal. Therefore, optimal limits for vitamin D administration need to be established for children and adolescents.

  3. 碳青霉烯类抗生素联合β-内酰胺/酶抑制剂治疗耐药肺炎克雷伯菌所致医院获得性肺炎疗效的回顾性分析%Retrospectively Analysis on the Clinical Effect of Carbapenems Combined with β - lactam/Enzyme Inhibitors on Hospital-acquired Pneumonia Caused by Drug-resistant Klebsiella Pneumoniae

    Institute of Scientific and Technical Information of China (English)

    赵世峰; 曹江涛; 冯丽洁

    2015-01-01

    Objective To retrospectively analyze the clinical effect of carbapenems combined with β - lactam/ enzyme inhibitors on hospital - acquired pneumonia caused by drug - resistant Klebsiella pneumoniae. Methods A total of 20 patients with hospital - acquired pneumonia caused by drug - resistant klebsiella pneumoniae were selected in General Hospital of the Chinese People's Liberation Army from June 2013 to December 2014,positive rate of bacterial strains with ESBLs and incidence of drug resistance were analyzed;all of the 20 patients received carbapenems combined with β - lactam/ enzyme inhibitors, clinical effect after 14 days of treatment,fatality rate after 30 days of treatment and incidence of adverse reactions during treatment were observed. Results A total of 14 bacterial strains with ESBLs were found( accounted for 70% ),the drug resistant rates of Klebsiella pneumoniae to penicillins, β - lactam/ enzyme inhibitors, carbapenems, aminoglycosides, quinolones and trimethoprim/ sulfamethoxazole were all over 70% . The effective rate was 65% , fatality rate was 35% , attribution mortality was 20% . During treatment,2 cases occurred mild nausea and vomiting,2 cases occurred rash,4 cases occurred intestinal double infection. Conclusion Carbapenems combined with β - lactam/ enzyme inhibitors is effective and safe in treating hospital - acquired pneumonia caused by drug - resistant Klebsiella pneumoniae.%目的:回顾性分析碳青霉烯类抗生素联合β-内酰胺/酶抑制剂治疗耐药肺炎克雷伯菌所致医院获得性肺炎(HAP)的疗效。方法选取2013年6月—2014年12月在解放军总医院确诊的耐药肺炎克雷伯菌所致 HAP 患者20例,分析其产超广谱β-内酰胺酶(产 ESBLs)菌株分布及耐药情况,并给予碳青霉烯类抗生素联合β-内酰胺/酶抑制剂治疗,观察治疗14 d 后临床疗效、治疗30 d 后病死率及治疗期间不良反应。结果20例患者中发现产 ESBLs菌株14例(占70%

  4. What Causes Hypotension?

    Science.gov (United States)

    ... enzyme (ACE) inhibitors Angiotensin II receptor blockers Nitrates Beta blockers Medicines for conditions such as anxiety, depression, erectile dysfunction, and central nervous system disorders ...

  5. Stability of Enzymes in Granular Enzyme Products for Laundry Detergents

    OpenAIRE

    Biran, Suzan; Jensen, Anker Degn; Kiil, Søren; Bach, Poul; Simonsen, Ole

    2010-01-01

    Enzymes have long been of interest to the detergent industry due to their ability to improve the cleaning efficiency of synthetic detergents, contribute to shortening washing times, and reduce energy and water consumption, provision of environmentally friendlier wash water effluents and fabric care. However, incorporating enzymes in detergent formulations gives rise to numerous practical problems due to their incompatibility with and stability against various detergent components. In powdered...

  6. GRE Enzymes for Vector Analysis

    Data.gov (United States)

    U.S. Environmental Protection Agency — Microbial enzyme data that were collected during the 2004-2006 EMAP-GRE program. These data were then used by Moorhead et al (2016) in their ecoenzyme vector...

  7. Controlled enzyme catalyzed heteropolysaccharide degradation

    DEFF Research Database (Denmark)

    Rasmussen, Louise Enggaard

    The work presented in this PhD thesis has provided a better understanding of the enzyme kinetics and quantitative phenomena of the hydrolysis of xylan substrates by selected pure enzyme preparations. Furthermore, the options for producing specific substituted xylooligosaccharides from selected...... substrates by specific xylanase treatment have been examined. The kinetics of the enzymatic degradation of water-extractable wheat arabinoxylan (WE-AX) during designed treatments with selected monocomponent enzymes was investigated by monitoring the release of xylose and arabinose. The results of different...... between -xylosidase and the α-L-arabinofuranosidases on the xylose release were low as compared to the effect of xylanase addition with β-xylosidase, which increased the xylose release by ~25 times in 30 minutes. At equimolar addition levels of the four enzymes, the xylanase activity was thus rate...

  8. A Review on the Effects of Supercritical Carbon Dioxide on Enzyme Activity

    Directory of Open Access Journals (Sweden)

    Marie Zarevúcka

    2010-01-01

    Full Text Available Different types of enzymes such as lipases, several phosphatases, dehydrogenases, oxidases, amylases and others are well suited for the reactions in SC-CO2. The stability and the activity of enzymes exposed to carbon dioxide under high pressure depend on enzyme species, water content in the solution and on the pressure and temperature of the reaction system. The three-dimensional structure of enzymes may be significantly altered under extreme conditions, causing their denaturation and consequent loss of activity. If the conditions are less adverse, the protein structure may be largely retained. Minor structural changes may induce an alternative active protein state with altered enzyme activity, specificity and stability.

  9. Enzymes: principles and biotechnological applications.

    Science.gov (United States)

    Robinson, Peter K

    2015-01-01

    Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in living organisms, and which can be extracted from cells and then used to catalyse a wide range of commercially important processes. This chapter covers the basic principles of enzymology, such as classification, structure, kinetics and inhibition, and also provides an overview of industrial applications. In addition, techniques for the purification of enzymes are discussed.

  10. Enzymes: principles and biotechnological applications

    Science.gov (United States)

    Robinson, Peter K.

    2015-01-01

    Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in living organisms, and which can be extracted from cells and then used to catalyse a wide range of commercially important processes. This chapter covers the basic principles of enzymology, such as classification, structure, kinetics and inhibition, and also provides an overview of industrial applications. In addition, techniques for the purification of enzymes are discussed. PMID:26504249

  11. Alleviating Cancer Drug Toxicity by Inhibiting a Bacterial Enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Wallace, Bret D.; Wang, Hongwei; Lane, Kimberly T.; Scott, John E.; Orans, Jillian; Koo, Ja Seol; Venkatesh, Madhukumar; Jobin, Christian; Yeh, Li-An; Mani, Sridhar; Redinbo, Matthew R. (Einstein); (UNC); (North Carolina Central University)

    2011-08-12

    The dose-limiting side effect of the common colon cancer chemotherapeutic CPT-11 is severe diarrhea caused by symbiotic bacterial {beta}-glucuronidases that reactivate the drug in the gut. We sought to target these enzymes without killing the commensal bacteria essential for human health. Potent bacterial {beta}-glucuronidase inhibitors were identified by high-throughput screening and shown to have no effect on the orthologous mammalian enzyme. Crystal structures established that selectivity was based on a loop unique to bacterial {beta}-glucuronidases. Inhibitors were highly effective against the enzyme target in living aerobic and anaerobic bacteria, but did not kill the bacteria or harm mammalian cells. Finally, oral administration of an inhibitor protected mice from CPT-11-induced toxicity. Thus, drugs may be designed to inhibit undesirable enzyme activities in essential microbial symbiotes to enhance chemotherapeutic efficacy.

  12. Application of an enzyme-labeled antigen method for visualizing plasma cells producing antibodies against Strep A, a carbohydrate antigen of Streptococcus pyogenes, in recurrent tonsillitis.

    Science.gov (United States)

    Onouchi, Takanori; Mizutani, Yasuyoshi; Shiogama, Kazuya; Inada, Ken-ichi; Okada, Tatsuyoshi; Naito, Kensei; Tsutsumi, Yutaka

    2015-01-01

    Streptococcus pyogenes is the main causative pathogen of recurrent tonsillitis. Histologically, lesions of recurrent tonsillitis contain numerous plasma cells. Strep A is an antigenic carbohydrate molecule on the cell wall of S. pyogenes. As expected, plasma cells in subjects with recurrent tonsillitis secrete antibodies against Strep A. The enzyme-labeled antigen method is a novel histochemical technique that visualizes specific antibody-producing cells in tissue sections by employing a biotin-labeled antigen as a probe. The purpose of the present study was to visualize plasma cells producing antibodies reactive with Strep A in recurrent tonsillitis. Firstly, the lymph nodes of rats immunized with boiled S. pyogenes were paraformaldehyde-fixed and specific plasma cells localized in frozen sections with biotinylated Strep A. Secondly, an enzyme-labeled antigen method was used on human tonsil surgically removed from 12 patients with recurrent tonsillitis. S. pyogenes genomes were PCR-detected in all 12 specimens. The emm genotypes belonged to emm12 in nine specimens and emm1 in three. Plasma cells producing anti-Strep A antibodies were demonstrated in prefixed frozen sections of rat lymph nodes, 8/12 human specimens from patients with recurrent tonsillitis but not in two control tonsils. In human tonsils, Strep A-reactive plasma cells were observed within the reticular squamous mucosa and just below the mucosa, and the specific antibodies belonged to either IgA or IgG classes. Our technique is effective in visualizing immunocytes producing specific antibodies against the bacterial carbohydrate antigen, and is thus a novel histochemical tool for analyzing immune reactions in infectious disorders.

  13. Engineering Cellulase Enzymes for Bioenergy

    Science.gov (United States)

    Atreya, Meera Elizabeth

    Sustainable energy sources, such as biofuels, offer increasingly important alternatives to fossil fuels that contribute less to global climate change. The energy contained within cellulosic biofuels derives from sunlight energy stored in the form of carbon-carbon bonds comprising sugars such as glucose. Second-generation biofuels are produced from lignocellulosic biomass feedstocks, including agricultural waste products and non-food crops like Miscanthus, that contain lignin and the polysaccharides hemicellulose and cellulose. Cellulose is the most abundant biological material on Earth; it is a polymer of glucose and a structural component of plant cell walls. Accessing the sugar is challenging, as the crystalline structure of cellulose resists degradation; biochemical and thermochemical means can be used to depolymerize cellulose. Cellulase enzymes catalyze the biochemical depolymerization of cellulose into glucose. Glucose can be used as a carbon source for growth of a biofuel-producing microorganism. When it converts glucose to a hydrocarbon fuel, this microbe completes the biofuels process of transforming sunlight energy into accessible, chemical energy capable of replacing non-renewable transportation fuels. Due to strong intermolecular interactions between polymer chains, cellulose is significantly more challenging to depolymerize than starch, a more accessible polymer of glucose utilized in first-generation biofuels processes (often derived from corn). While most mammals cannot digest cellulose (dietary fiber), certain fungi and bacteria produce cellulase enzymes capable of hydrolyzing it. These organisms secrete a wide variety of glycoside hydrolase and other classes of enzymes that work in concert. Because cellulase enzymes are slow-acting and expensive to produce, my aim has been to improve the properties of these enzymes as a means to make a cellulosic biofuels process possible that is more efficient and, consequently, more economical than current

  14. Histochemical and ultrastructural study of Caesalpinia peltophoroides Benth. (Leguminosae-Caesalpinoideae seeds Estudo histoquímico e ultraestrutural de sementes de Caesalpinia peltophoroides Benth. (Leguminosae-Caesalpinoideae

    Directory of Open Access Journals (Sweden)

    Viviana Borges Corte

    2009-10-01

    Full Text Available The objective of this work was to correlate data on light microscopy observations through histochemical analysis and polarized light techniques and investigations in transmission electron microscopy (TEM to characterize the reserve materials in C. peltophoroides Benth. (Leguminosae-Caesalpinoideae cotyledons, popularly known as "sibipiruna", a tropical tree species with wide distribution in Brazil. The cotyledon mesophyll, especially in the abaxial face, is rich in unsaturated neutral lipids contained in numerous lipid bodies dispersed in the cytoplasm. Proteins, more concentrated in the adaxial face of the cotyledons, occur in all the mesophyll and are stored in protein bodies containing globoids, with variable number and size, responsible for accumulation of mineral reserves. Calcium oxalate druses have distribution restricted to the cotyledons adaxial face and are associated with protein bodies. Starch, also distributed all over the cotyledon mesophyll, occurs in small amounts in plastids with developed lamellar system. Secretory cavities rich in phenolic compounds occur among procambial strands.Este trabalho procurou correlacionar dados de observações em microscopia de luz através de técnicas histoquímicas e de luz polarizada e investigações em microscopia eletrônica de transmissão para caracterizar os materiais de reserva em cotilédones de C. peltophoroides Benth. (Leguminosae-Caesalpinoideae, conhecida popularmente como sibipiruna, ou falso pau-brasil, uma espécie tropical com ampla distribuição no Brasil. O mesofilo cotiledonar, especialmente na face abaxial, apresenta-se rico em lipídios neutros, insaturados, contidos em numerosos corpos lipídicos dispersos no citoplasma. As proteínas, mais concentradas na face adaxial dos cotilédones, ocorrem em todo o mesofilo e são armazenadas em corpos proteicos contendo globoides, com número e tamanho variáveis, responsáveis pelo acúmulo de reservas minerais. Drusas de oxalato

  15. Functional electrical stimulation as a safe and effective treatment for equine epaxial muscle spasms: Clinical evaluations and histochemical morphometry of mitochondria in muscle biopsies

    Directory of Open Access Journals (Sweden)

    Barbara Ravara

    2015-03-01

    Full Text Available Functional Electrical Stimulation (FES has been used extensively over several decades to reverse muscle atrophy during rehabilitation for spinal cord injury patients. The benefits of the technology are being expanded into other areas, and FES has been recently utilized for injury rehabilitation and performance enhancement in horses. Six retired horses (age from 10 to 17 yrs that had been previously used mainly for dressage riding were selected for this study. Clinical evaluation found epaxial muscle spasms in all horses with minimal to no pelvic extension when manually palpated. FES treatments were performed on the sacral/lumbar region 3 times per week for a period of 8 weeks, obtaining a total of 22 treatments per horse. The Modified Ashworth Scale for grading muscle spasms found a one grade improvement after approximately four FES treatments, indicating improved functional movement of the sacral/lumbar region, supporting the evidence by clinical palpations that a reduction in epaxial muscle spasms occurred. Skeletal muscle biopsies Pre and Post FES treatments were obtained from the longissimus lumborum muscle. Cryosections were stained with a Hemotoxylin-Eosin (H-E, and nicotinamide adenine dinucleotide tetrazolium reductase reaction (NADH-TR. The eventual size change of the muscle fibers were evaluated by morphometry in the H-E and NADH-TR stained cryosections, while in the NADH-TR slides the histochemical density and distribution of mitochondria were also determined. The main results of the morphometric analyses were: 1 As expected for the type of FES treatment used in this study, only a couple of horses showed significant increases in mean muscle fiber size when Pre- vs Post-FES biopsies were compared; 2 In the older horses, there were sparse (or many in one horse very atrophic and angulated muscle fibers in both Pre- and Post-FES samples, whose attributes and distribution suggests that they were denervated due to a distal neuropathy; 3 The

  16. Nemaline myopathy: clinical, histochemical and immunohistochemical features Miopatia nemalínica: achados clínicos, histoquímicos e imuno-histoquímicos

    Directory of Open Access Journals (Sweden)

    Nazah Cherif Mohamad Youssef

    2009-09-01

    Full Text Available Nemaline myopathy (NM is a congenital disease that leads to hypotonia and feeding difficulties in neonates. Some cases have a more benign course, with skeletal abnormalities later in life. We analyzed a series of eight patients with NM obtained from a retrospective analysis of 4300 muscle biopsies. Patients were classified as having the typical form in five cases, intermediate form in two cases and severe form in one case. Histochemical analysis showed mixed rods distribution in all cases and predominance of type I fibers in five cases. Immunohistochemical analysis showed abnormal nebulin expression in all patients (four heterogeneous and four absent, homogeneous desmin expression in four cases, strongly positive in three and absent in one, fast myosin expression in a mosaic pattern in six cases and absent in two cases. There was no specific relation between these protein expression patterns and the clinical forms of NM.Miopatia nemalínica (NM é uma doença congênita que leva a hipotonia e dificuldade de sugar em neonatos. Alguns casos possuem uma evolução benigna, com deformidades ósseas tardias. Nós analisamos uma série de oito pacientes com NM obtidos da análise retrospectiva de 4300 biópsias musculares. Os pacientes foram classificados como forma típica em cinco casos, forma intermediária em dois casos e forma severa em um caso. Análise histoquímica mostrou distribuição mista dos rods em todos os casos e predominância de fibras tipo I em cinco casos. Análise imuno-histoquímica mostrou expressão anormal da nebulina em todos os pacientes (quatro heterogênea e quatro ausente, expressão homogenea da desmina em quatro casos, fortemente positiva em tres e ausente em um, expressão da miosina (rápida com padrão em mosaico em seis casos e ausente em dois casos. Não há relação específica entre a expressão destas proteínas e as formas clínicas da NM.

  17. Heavy enzymes--experimental and computational insights in enzyme dynamics.

    Science.gov (United States)

    Swiderek, Katarzyna; Ruiz-Pernía, J Javier; Moliner, Vicent; Tuñón, Iñaki

    2014-08-01

    The role of protein motions in the chemical step of enzyme-catalyzed reactions is the subject of an open debate in the scientific literature. The systematic use of isotopically substituted enzymes has been revealed as a useful tool to quantify the role of these motions. According to the Born-Oppenheimer approximation, changing the mass of the protein does not change the forces acting on the system but alters the frequencies of the protein motions, which in turn can affect the rate constant. Experimental and theoretical studies carried out in this field are presented in this article and discussed in the framework of Transition State Theory.

  18. The Kinetics of Enzyme Mixtures

    Directory of Open Access Journals (Sweden)

    Simon Brown

    2014-03-01

    Full Text Available Even purified enzyme preparations are often heterogeneous. For example, preparations of aspartate aminotransferase or cytochrome oxidase can consist of several different forms of the enzyme. For this reason we consider how different the kinetics of the reactions catalysed by a mixture of forms of an enzyme must be to provide some indication of the characteristics of the species present. Based on the standard Michaelis-Menten model, we show that if the Michaelis constants (Km of two isoforms differ by a factor of at least 20 the steady-state kinetics can be used to characterise the mixture. However, even if heterogeneity is reflected in the kinetic data, the proportions of the different forms of the enzyme cannot be estimated from the kinetic data alone. Consequently, the heterogeneity of enzyme preparations is rarely reflected in measurements of their steady-state kinetics unless the species present have significantly different kinetic properties. This has two implications: (1 it is difficult, but not impossible, to detect molecular heterogeneity using kinetic data and (2 even when it is possible, a considerable quantity of high quality data is required.

  19. Enzymes in CO2 Capture

    DEFF Research Database (Denmark)

    Fosbøl, Philip Loldrup; Gladis, Arne; Thomsen, Kaj

    of carbon capture is the application of enzymes for acceleration of typically slow ternary amines or inorganic carbonates. There is a hidden potential to revive currently infeasible amines which have an interesting low energy consumption for regeneration but too slow kinetics for viable CO2 capture. The aim......The enzyme Carbonic Anhydrase (CA) can accelerate the absorption rate of CO2 into aqueous solutions by several-fold. It exist in almost all living organisms and catalyses different important processes like CO2 transport, respiration and the acid-base balances. A new technology in the field...... of this work is to discuss the measurements of kinetic properties for CA promoted CO2 capture solvent systems. The development of a rate-based model for enzymes will be discussed showing the principles of implementation and the results on using a well-known ternary amine for CO2 capture. Conclusions...

  20. Expanding role of microsomal enzyme induction, and its implications for clinical chemistry

    Energy Technology Data Exchange (ETDEWEB)

    Goldberg, D.M.

    1980-05-01

    Microsomal enzyme induction, a term denoting the ability of the substrate for a microsomal enzyme to enhance the activity of that enzyme and frequently of related enzymes, has been demonstrated in a wide range of tissues, notably the liver, placenta, small intestinal muccosa, and peripheral lymphocytes. The major agents that cause microsomal enzyme induction are drugs and xenobiotics. Factors modulating the extent of enzyme induction by a given agent include age and nutrition, and wide species variations are encountered with different inducing agents. Markers for microsomal enzyme induction include determination of the plasma half-life for conveniently measured drugs, and the measurement of endogenous metabolites such as 6;-hydroxycortisol and D-glucaric acid in 24-h urine collections. While these are valuable for monitoring enzyme induction in healthy patients, they are altered in certain forms of liver disease, and results must then be interpreted with caution. Microsomal enzyme induction may interfere with reference values, particularly for membrane-bound enzymes, in otherwise healthy populations, and may play a role in metabolic bone disease, drug interactions, carcinogenesis, and hypertriglyceridemia. Drug therapy of the neonatal and congenital hyperbilirubinemias has been inspired by the mechanism of hepatic microsomal enzyme induction, and ''markers'' for enzyme induction can be used to monitor drug compliance. The activity of serum <-glutamyltransferase seems to be especially valuable for this purpose.

  1. Enzyme and biochemical producing fungi

    DEFF Research Database (Denmark)

    Lübeck, Peter Stephensen; Lübeck, Mette; Nilsson, Lena

    2010-01-01

    We are developing a biorefinery concept for biological production of chemicals, drugs, feed and fuels using plant biomass as raw material in well-defined cell-factories. Among the important goals is the discovery of new biocatalysts for production of enzymes, biochemicals and fuels and already our...... screening of a large collection of fungal strains isolated from natural habitats have resulted in identification of strains with high production of hydrolytic enzymes and excretion of organic acids. Our research focuses on creating a fungal platform based on synthetic biology for developing new cell...

  2. Enzyme inactivation in food processing using high pressure carbon dioxide technology.

    Science.gov (United States)

    Hu, Wanfeng; Zhou, Linyan; Xu, Zhenzhen; Zhang, Yan; Liao, Xiaojun

    2013-01-01

    High pressure carbon dioxide (HPCD) is an effective non-thermal processing technique for inactivating deleterious enzymes in liquid and solid food systems. This processing method avoids high temperatures and exerts a minimal impact on the nutritional and sensory properties of foods, but extends shelf life by inhibiting or killing microorganisms and enzymes. Indigenous enzymes in food such as polyphenol oxidase (PPO), pectin methylesterase (PME), and lypoxygenase (LOX) may cause undesirable chemical changes in food attributes, showing the loss in color, texture, and flavor. For more than two decades, HPCD has proved its effectiveness in inactivating these enzymes. The HPCD-induced inactivation of some microbial enzymes responsible for microbial metabolism is also included. This review presents a survey of the published knowledge regarding the use of HPCD for the inactivation of these enzymes, and analyzes the factors controlling the efficiency of HPCD and speculates on the underlying mechanism that leads to enzyme inactivation.

  3. Taking the Mystery Out of Enzymes.

    Science.gov (United States)

    DeYoung, H. Garrett

    1984-01-01

    Discusses structure and function of enzymes, design of new enzymes and enzyme substitutes, and enzyme uses in industry, medicine, and wastewater treatment. The latter is a low-cost method which can remove as much as 99 percent of toxic substances found in many industrial wastewater streams. (JN)

  4. What Causes Thyroid Cancer?

    Science.gov (United States)

    ... Cancer Causes, Risk Factors, and Prevention What Causes Thyroid Cancer? Thyroid cancer is linked with a number of ... inside a cell, without an outside cause. Papillary thyroid cancer Several DNA mutations (changes) have been found in ...

  5. What Causes a Toothache?

    Science.gov (United States)

    ... directly to your desktop! more... What Causes a Toothache? Article Chapters What Causes a Toothache? Why Does ... bite. Tooth eruption may be the cause of tooth or jaw pain in babies and school-age children. TMJ, sinus ...

  6. Enzyme nanoassemblies for biomass conversion

    Science.gov (United States)

    Biomass represents a vast resource for the production of the world’s fuel and chemical feedstock needs. The use of enzymes to effect these bioconversions offers an alternative that is potentially more specific and environmentally-friendly than harsher chemical methodologies. Some species of anaero...

  7. Insolubilized enzymes for food synthesis

    Science.gov (United States)

    Marshall, D. L.

    1972-01-01

    Cellulose matrix with numerous enzyme-coated silica particles of colloidal size permanently bound at various sites within matrix was produced that has high activity and possesses requisite physical characteristics for filtration or column operations. Product also allows coupling step in synthesis of edible food to proceed under mild conditions.

  8. The enzymes associated with denitrification

    Science.gov (United States)

    Hochstein, L. I.; Tomlinson, G. A.

    1988-01-01

    The enzymes involved in the reduction of nitrogenous oxides are thought to be intermediates in denitrification processes. This review examines the roles of nitrate reductase, nitrite reductases, nitric oxide reductase, mechanisms of N-N bond formation, and nitrous oxide reductases.

  9. Silica-Immobilized Enzyme Reactors

    Science.gov (United States)

    2007-08-01

    immobilized artificial membrane chromatography and lysophospholipid micellar electrokinetic chromatography . J. Chromatogr. A 1998, 810, 95-103. 50...Journal of Liquid Chromatography and Related Technologies. Air Force Research Laboratory Materials and Manufacturing Directorate Airbase...immobilized enzyme reactors (IMERs) can also be integrated directly to further analytical methods such as liquid chromatography or mass spectrometry.[6] In

  10. Rapid-Equilibrium Enzyme Kinetics

    Science.gov (United States)

    Alberty, Robert A.

    2008-01-01

    Rapid-equilibrium rate equations for enzyme-catalyzed reactions are especially useful because if experimental data can be fit by these simpler rate equations, the Michaelis constants can be interpreted as equilibrium constants. However, for some reactions it is necessary to use the more complicated steady-state rate equations. Thermodynamics is…

  11. Kathepsine C : Een allosterisch enzyme

    NARCIS (Netherlands)

    Gorter, Jeannette

    1969-01-01

    In chapter I an introduction into allosteric systems is given. In chapter II is a detailed method is described for the applica of Gly-Phe--p. nitroanilide (GPNA) as a substrate for the activity assay of the lysosomal enzyme cathepsin C. It is an allosteric which is activated by Cl-, Br-, 1-, CNS-, N

  12. Enzyme recovery using reversed micelles.

    NARCIS (Netherlands)

    Dekker, M.

    1990-01-01

    The objective of this study was to develop a liquid-liquid extraction process for the recovery of extracellular enzymes. The potentials of reaching this goal by using reversed micelles in an organic solvent have been investigated.Reversed micelles are aggregates of surfactant molecules containing an

  13. Fat malabsorption in cystic fibrosis patients receiving enzyme replacement therapy is due to impaired intestinal uptake of long-chain fatty acids

    NARCIS (Netherlands)

    Kalivianakis, M; Minich, DM; Bijleveld, CMA; van Aalderen, WMC; Stellaard, F; Vonk, RJ; Verkade, HJ

    1999-01-01

    Background: Pancreatic enzyme replacement therapy frequently fails to correct intestinal fat malabsorption completely in cystic fibrosis (CF) patients. The reason for this failure is unknown. Objective: We investigated whether fat malabsorption in CF patients treated with pancreatic enzymes is cause

  14. Inhibitors of Nucleotidyltransferase Superfamily Enzymes Suppress Herpes Simplex Virus Replication

    OpenAIRE

    2014-01-01

    Herpesviruses are large double-stranded DNA viruses that cause serious human diseases. Herpesvirus DNA replication depends on multiple processes typically catalyzed by nucleotidyltransferase superfamily (NTS) enzymes. Therefore, we investigated whether inhibitors of NTS enzymes would suppress replication of herpes simplex virus 1 (HSV-1) and HSV-2. Eight of 42 NTS inhibitors suppressed HSV-1 and/or HSV-2 replication by >10-fold at 5 μM, with suppression at 50 μM reaching ∼1 million-fold. Five...

  15. Thermodynamics of Enzyme-Catalyzed Reactions Database

    Science.gov (United States)

    SRD 74 Thermodynamics of Enzyme-Catalyzed Reactions Database (Web, free access)   The Thermodynamics of Enzyme-Catalyzed Reactions Database contains thermodynamic data on enzyme-catalyzed reactions that have been recently published in the Journal of Physical and Chemical Reference Data (JPCRD). For each reaction the following information is provided: the reference for the data, the reaction studied, the name of the enzyme used and its Enzyme Commission number, the method of measurement, the data and an evaluation thereof.

  16. Virulence-Associated Enzymes of Cryptococcus neoformans

    OpenAIRE

    2015-01-01

    Enzymes play key roles in fungal pathogenesis. Manipulation of enzyme expression or activity can significantly alter the infection process, and enzyme expression profiles can be a hallmark of disease. Hence, enzymes are worthy targets for better understanding pathogenesis and identifying new options for combatting fungal infections. Advances in genomics, proteomics, transcriptomics, and mass spectrometry have enabled the identification and characterization of new fungal enzymes. This review f...

  17. Degradation of various dyes using Laccase enzyme.

    Science.gov (United States)

    Dhaarani, S; Priya, A K; Rajan, T Vel; Kartic, D Navamani

    2012-10-01

    Disposal of untreated dyeing effluent in water bodies, from textile industries, cause serious environmental and health hazards. The chemical structures of dye molecules are designed to resist fading on exposure to light or chemical attack, and they prove to be quite resistant towards microbial degradation. Therefore, current conventional biological processes may not be able to meet wastewater discharge criteria and reuse. An enzymatic treatment undergoes oxidative cleavage avoiding formation of toxic amines. Laccase is a multi-copper containing protein that catalyzes the oxidation of a wide range of aromatic substrates concomitantly with the reduction of molecular oxygen to water. UV visible spectral analysis of various synthetic dyes was performed in the study and wavelengths of maximum absorbance determined. Laccase enzyme was obtained from the fungi Pleorotus ostreatus. The enzyme showed high efficiency against Malachite Green, Basic Red and Acid Majanta with decolorization capacities of 97%, 94% and 94% respectively. Further, these dyes can be used for optimization of degradation parameters and analysis of degradation products.

  18. Enzymes involved in triglyceride hydrolysis.

    Science.gov (United States)

    Taskinen, M R; Kuusi, T

    1987-08-01

    The lipolytic enzymes LPL and HL play important roles in the metabolism of lipoproteins and participate in lipoprotein interconversions. LPL was originally recognized to be the key enzyme in the hydrolysis of chylomicrons and triglyceride, but it also turned out to be one determinant of HDL concentration in plasma. When LPL activity is high, chylomicrons and VLDL are rapidly removed from circulation and a concomitant rise of the HDL2 occurs. In contrast, low LPL activity impedes the removal of triglyceride-rich particles, resulting in the elevation of serum triglycerides and a decrease of HDL (HDL2). Concordant changes of this kind in LPL and HDL2 are induced by many physiological and pathological perturbations. Finally, the operation of LPL is also essential for the conversion of VLDL to LDL. This apparently clear-cut role of LPL in lipoprotein interconversions is contrasted with the enigmatic actions of HL. The enzyme was originally thought to participate in the catalyses of chylomicron and VLDL remnants generated in the LPL reaction. However, substantial in vitro and in vivo data indicate that HL is a key enzyme in the degradation of plasma HDL (HDL2) in a manner which opposes LPL. A scheme is presented for the complementary actions of the two enzymes in plasma HDL metabolism. In addition, recent studies have attributed a role to HL in the catabolism of triglyceride-rich lipoproteins, particularly those containing apo E. However, this function becomes clinically important only under conditions where the capacity of the LPL-mediated removal system is exceeded. Such a situation may arise when the input of triglyceride-rich particles (chylomicrons and/or VLDL) is excessive or LPL activity is decreased or absent.

  19. Platelet enzyme abnormalities in leukemias

    Directory of Open Access Journals (Sweden)

    S Sharma

    2011-01-01

    Full Text Available Aim of the Study: The aim of this study was to evaluate platelet enzyme activity in cases of leukemia. Materials and Methods: Platelet enzymes glucose-6-phosphate dehydrogenase (G6PD, pyruvate kinase (PK and hexokinase (HK were studied in 47 patients of acute and chronic leukemia patients, 16 patients with acute myeloid leukemia (AML(13 relapse, three in remission, 12 patients with acute lymphocytic leukemia (ALL (five in relapse, seven in remission, 19 patients with chronic myeloid leukemia (CML. Results: The platelet G6PD activity was significantly low in cases of AML, ALL and also in CML. G6PD activity was normalized during AML remission. G6PD activity, although persistently low during ALL remission, increased significantly to near-normal during remission (P < 0.05 as compared with relapse (P < 0.01. Platelet PK activity was high during AML relapse (P < 0.05, which was normalized during remission. Platelet HK however was found to be decreased during all remission (P < 0.05. There was a significant positive correlation between G6PD and PK in cases of AML (P < 0.001 but not in ALL and CML. G6PD activity did not correlate with HK activity in any of the leukemic groups. A significant positive correlation was however seen between PK and HK activity in cases of ALL remission (P < 0.01 and CML (P < 0.05. Conclusions: Both red cell and platelet enzymes were studied in 36 leukemic patients and there was no statistically significant correlation between red cell and platelet enzymes. Platelet enzyme defect in leukemias suggests the inherent abnormality in megakaryopoiesis and would explain the functional platelet defects in leukemias.

  20. Engineering cytochrome p450 enzymes.

    Science.gov (United States)

    Gillam, Elizabeth M J

    2008-01-01

    The last 20 years have seen the widespread and routine application of methods in molecular biology such as molecular cloning, recombinant protein expression, and the polymerase chain reaction. This has had implications not only for the study of toxicological mechanisms but also for the exploitation of enzymes involved in xenobiotic clearance. The engineering of P450s has been performed with several purposes. The first and most fundamental has been to enable successful recombinant expression in host systems such as bacteria. This in turn has led to efforts to solubilize the proteins as a prerequisite to crystallization and structure determination. Lagging behind has been the engineering of enzyme activity, hampered in part by our still-meager comprehension of fundamental structure-function relationships in P450s. However, the emerging technique of directed evolution holds promise in delivering both engineered enzymes for use in biocatalysis and incidental improvements in our understanding of sequence-structure and sequence-function relationships, provided that data mining can extract the fundamental correlations underpinning the data. From the very first studies on recombinant P450s, efforts were directed toward constructing fusions between P450s and redox partners in the hope of generating more efficient enzymes. While this aim has been allowed to lie fallow for some time, this area merits further investigation as does the development of surface-displayed P450 systems for biocatalytic and biosensor applications. The final application of engineered P450s will require other aspects of their biology to be addressed, such as tolerance to heat, solvents, and high substrate and product concentrations. The most important application of these enzymes in toxicology in the near future is likely to be the biocatalytic generation of drug metabolites for the pharmaceutical industry. Further tailoring will be necessary for specific toxicological applications, such as in

  1. Cerebrovascular effects of angiotensin converting enzyme inhibition involve large artery dilatation in rats

    DEFF Research Database (Denmark)

    Postiglione, A; Bobkiewicz, T; Vinholdt-Pedersen, E

    1991-01-01

    The aim of the study was to selectively examine the effects of converting enzyme inhibition on the large brain arteries by using concomitant inhibition of carbonic anhydrase to cause severe dilatation of mainly parenchymal resistance vessels....

  2. 7 CFR 58.436 - Rennet, pepsin, other milk clotting enzymes and flavor enzymes.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Rennet, pepsin, other milk clotting enzymes and flavor enzymes. 58.436 Section 58.436 Agriculture Regulations of the Department of Agriculture (Continued... clotting enzymes and flavor enzymes. Enzyme preparations used in the manufacture of cheese shall be...

  3. Mitochondrial myopathy caused by long-term zidovudine therapy.

    Science.gov (United States)

    Dalakas, M C; Illa, I; Pezeshkpour, G H; Laukaitis, J P; Cohen, B; Griffin, J L

    1990-04-19

    Both infection with the human immunodeficiency virus type 1 (HIV) and zidovudine (formerly called azidothymidine [AZT]) cause myopathy. To identify criteria for distinguishing zidovudine-induced myopathy from that caused by primary HIV infection, we reviewed the histochemical, immunocytochemical, and electron-microscopical features of muscle-biopsy specimens from 20 HIV-positive patients with myopathy (15 of whom had been treated with zidovudine) and compared the findings with the patients' clinical course and response to various therapies. Among the zidovudine-treated patients, the myopathy responded to prednisone in four, to the discontinuation of zidovudine in eight, and to nonsteroidal anti-inflammatory drugs in two. Numerous "ragged-red" fibers, indicative of abnormal mitochondria with paracrystalline inclusions, were found in the biopsy specimens from the zidovudine-treated patients but not in those from the other patients. The number of these fibers appeared to correlate with the severity of the myopathy. All the patients, regardless of whether they had been treated with zidovudine, had inflammatory myopathy characterized by degenerating fibers, cytoplasmic bodies, and endomysial infiltrates consisting of CD8+ cells (mean +/- SD, 60.7 +/- 6.4 percent) and macrophages (39.2 +/- 6.4 percent) associated with Class I major histocompatibility complex (MHC-I) antigens (HLA-A, -B, and -C antigens) in the muscle fibers. The numbers and percentages of CD8+ cells and macrophages were similar in both the zidovudine-treated and the untreated HIV-positive patients. Specimens obtained on repeat muscle biopsy from two patients in whom the myopathy responded to the discontinuation of zidovudine showed remarkable histologic improvement. We conclude that long-term therapy with zidovudine can cause a toxic mitochondrial myopathy, which coexists with a T-cell-mediated inflammatory myopathy that is restricted to MHC-I antigen, and is indistinguishable from the myopathy

  4. Enzyme technology: Key to selective biorefining

    DEFF Research Database (Denmark)

    Meyer, Anne S.

    2014-01-01

    to the reaction is a unique trait of enzyme catalysis. Since enzyme selectivity means that a specific reaction is catalysed between particular species to produce definite products, enzymes are particularly fit for converting specific compounds in mixed biomass streams. Since enzymes are protein molecules...... their rational use in biorefinery processes requires an understanding of the basic features of enzymes and reaction traits with respect to specificity, kinetics, reaction optima, stability and structure-function relations – we are now at a stage where it is possible to use nature’s enzyme structures as starting...... point and then improve the functional traits by targeted mutation of the protein. The talk will display some of our recent hypotheses related to enzyme action, recently obtained results within knowledge-based enzyme improvements as well as cast light on research methods used in optimizing enzyme...

  5. Petroleum-Degrading Enzymes: Bioremediation and New Prospects

    Directory of Open Access Journals (Sweden)

    R. S. Peixoto

    2011-01-01

    Full Text Available Anthropogenic forces, such as petroleum spills and the incomplete combustion of fossil fuels, have caused an accumulation of petroleum hydrocarbons in the environment. The accumulation of petroleum and its derivatives now constitutes an important environmental problem. Biocatalysis introduces new ways to improve the development of bioremediation strategies. The recent application of molecular tools to biocatalysis may improve bioprospecting research, enzyme yield recovery, and enzyme specificity, thus increasing cost-benefit ratios. Enzymatic remediation is a valuable alternative as it can be easier to work with than whole organisms, especially in extreme environments. Furthermore, the use of free enzymes avoids the release of exotic or genetically modified organisms (GMO in the environment.

  6. Skin symptoms in patients with atopic dermatitis using enzyme-containing detergents

    DEFF Research Database (Denmark)

    Andersen, Peter Hundevadt; Bindslev-Jensen, C; Mosbech, H

    1998-01-01

    Detergent enzymes may cause skin irritation and occasionally hypersensitivity reactions. The potential hazards of these enzymes have led some physicians to advise atopic dermatitis patients against the use of enzyme-enriched detergents. A three-phased randomised, double-blind, cross-over experiment...... differences in any of the primary or secondary parameters comparing treatment and placebo periods. Our data therefore seem to exclude that atopic dermatitis may exacerbate during 1 month's exposure to enzyme-enriched detergents. Since no significant irritant capacity was detected in atopic dermatitis patients...

  7. Finding homes for orphan enzymes

    Directory of Open Access Journals (Sweden)

    Frank M. Raushel

    2016-12-01

    Full Text Available The rate at which new genes are being sequenced greatly exceeds our ability to correctly annotate the functional properties of the corresponding proteins. Annotations based primarily on sequence identity to experimentally characterized proteins are often misleading because closely related sequences may have different functions, while highly divergent sequences may have identical functions. Our understanding of the principles that dictate the catalytic properties of enzymes, based on protein sequence alone, is often insufficient to correctly annotate proteins of unknown function. To address these problems, we are working to develop a comprehensive strategy for the functional annotation of newly sequenced genes using a combination of structural biology, bioinformatics, computational biology, and molecular enzymology. The power of this multidisciplinary approach for discovering new reactions catalyzed by uncharacterized enzymes has been tested using the amidohydrolase superfamily as a model system.

  8. Multi-enzyme Process Modeling

    DEFF Research Database (Denmark)

    Andrade Santacoloma, Paloma de Gracia

    . In this way the model parameters that drives the main dynamic behavior can be identified and thus a better understanding of this type of processes. In order to develop, test and verify the methodology, three case studies were selected, specifically the bi-enzyme process for the production of lactobionic acid......The subject of this thesis is to develop a methodological framework that can systematically guide mathematical model building for better understanding of multi-enzyme processes. In this way, opportunities for process improvements can be identified by analyzing simulations of either existing...... in the scientific literature. Reliable mathematical models of such multi-catalytic schemes can exploit the potential benefit of these processes. In this way, the best outcome of the process can be obtained understanding the types of modification that are required for process optimization. An effective evaluation...

  9. Improvements of biomass deconstruction enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Sale, K. L.

    2012-03-01

    Sandia National Laboratories and DSM Innovation, Inc. collaborated on the investigation of the structure and function of cellulases from thermophilic fungi. Sandia's role was to use its expertise in protein structure determination and X-ray crystallography to solve the structure of these enzymes in their native state and in their substrate and product bound states. Sandia was also tasked to work with DSM to use the newly solved structure to, using computational approaches, analyze enzyme interactions with both bound substrate and bound product; the goal being to develop approaches for rationally designing improved cellulases for biomass deconstruction. We solved the structures of five cellulases from thermophilic fungi. Several of these were also solved with bound substrate/product, which allowed us to predict mutations that might enhance activity and stability.

  10. Enzyme dynamics from NMR spectroscopy.

    Science.gov (United States)

    Palmer, Arthur G

    2015-02-17

    CONSPECTUS: Biological activities of enzymes, including regulation or coordination of mechanistic stages preceding or following the chemical step, may depend upon kinetic or equilibrium changes in protein conformations. Exchange of more open or flexible conformational states with more closed or constrained states can influence inhibition, allosteric regulation, substrate recognition, formation of the Michaelis complex, side reactions, and product release. NMR spectroscopy has long been applied to the study of conformational dynamic processes in enzymes because these phenomena can be characterized over multiple time scales with atomic site resolution. Laboratory-frame spin-relaxation measurements, sensitive to reorientational motions on picosecond-nanosecond time scales, and rotating-frame relaxation-dispersion measurements, sensitive to chemical exchange processes on microsecond-millisecond time scales, provide information on both conformational distributions and kinetics. This Account reviews NMR spin relaxation studies of the enzymes ribonuclease HI from mesophilic (Escherichia coli) and thermophilic (Thermus thermophilus) bacteria, E. coli AlkB, and Saccharomyces cerevisiae triosephosphate isomerase to illustrate the contributions of conformational flexibility and dynamics to diverse steps in enzyme mechanism. Spin relaxation measurements and molecular dynamics (MD) simulations of the bacterial ribonuclease H enzymes show that the handle region, one of three loop regions that interact with substrates, interconverts between two conformations. Comparison of these conformations with the structure of the complex between Homo sapiens ribonuclease H and a DNA:RNA substrate suggests that the more closed state is inhibitory to binding. The large population of the closed conformation in T. thermophilus ribonuclease H contributes to the increased Michaelis constant compared with the E. coli enzyme. NMR spin relaxation and fluorescence spectroscopy have characterized a

  11. Substrate mediated enzyme prodrug therapy.

    Directory of Open Access Journals (Sweden)

    Betina Fejerskov

    Full Text Available In this report, we detail Substrate Mediated Enzyme Prodrug Therapy (SMEPT as a novel approach in drug delivery which relies on enzyme-functionalized cell culture substrates to achieve a localized conversion of benign prodrug(s into active therapeutics with subsequent delivery to adhering cells or adjacent tissues. For proof-of-concept SMEPT, we use surface adhered micro-structured physical hydrogels based on poly(vinyl alcohol, β-glucuronidase enzyme and glucuronide prodrugs. We demonstrate enzymatic activity mediated by the assembled hydrogel samples and illustrate arms of control over rate of release of model fluorescent cargo. SMEPT was not impaired by adhering cells and afforded facile time - and dose - dependent uptake of the in situ generated fluorescent cargo by hepatic cells, HepG2. With the use of a glucuronide derivative of an anticancer drug, SN-38, SMEPT afforded a decrease in cell viability to a level similar to that achieved using parent drug. Finally, dose response was achieved using SMEPT and administration of judiciously chosen concentration of SN-38 glucuronide prodrug thus revealing external control over drug delivery using drug eluting surface. We believe that this highly adaptable concept will find use in diverse biomedical applications, specifically surface mediated drug delivery and tissue engineering.

  12. Characterization of CIM monoliths as enzyme reactors.

    Science.gov (United States)

    Vodopivec, Martina; Podgornik, Ales; Berovic, Marin; Strancar, Ales

    2003-09-25

    The immobilization of the enzymes citrate lyase, malate dehydrogenase, isocitrate dehydrogenase and lactate dehydrogenase to CIM monolithic supports was performed. The long-term stability, reproducibility, and linear response range of the immobilized enzyme reactors were investigated along with the determination of the kinetic behavior of the enzymes immobilized on the CIM monoliths. The Michaelis-Menten constant K(m) and the turnover number k(3) of the immobilized enzymes were found to be flow-unaffected. Furthermore, the K(m) values of the soluble and immobilized enzyme were found to be comparable. Both facts indicate the absence of a diffusional limitation in immobilized CIM enzyme reactors.

  13. Lignolytic Enzymes Production from Selected Mushrooms

    Directory of Open Access Journals (Sweden)

    H.M. Shantaveera Swamy

    2015-06-01

    Full Text Available In this paper, ligninase enzymes produced by selected mushrooms have been reported. We collected mushrooms from Western Ghats, most of them were edible food. Thirty samples isolated were tested using a plate assay through direct agar plate assay by using ABTS, decolourisation containing the fifteen isolates were able to decolourise the dye, indicating a lignin-degrading ability. Spectrophotometric enzyme assays from all selected isolates were carried out to examine the production of Ligninolytic enzymes (Laccase, lignin peroxidase and manganese peroxidase. Ten selected isolates produced all three kinds of enzymes tested. Lignolytic enzymes are groups of enzymes these are actively involved in bioremediation.

  14. Distal weakness with respiratory insufficiency caused by the m.8344A > G "MERRF" mutation.

    Science.gov (United States)

    Blakely, Emma L; Alston, Charlotte L; Lecky, Bryan; Chakrabarti, Biswajit; Falkous, Gavin; Turnbull, Douglass M; Taylor, Robert W; Gorman, Grainne S

    2014-06-01

    The m.8344A>G mutation in the mt-tRNA(Lys) gene, first described in myoclonic epilepsy and ragged red fibers (MERRF), accounts for approximately 80% of mutations in individuals with MERRF syndrome. Although originally described in families with a classical syndrome of myoclonus, ataxia, epilepsy and ragged red fibers in muscle biopsy, the m.8344A>G mutation is increasingly recognised to exhibit marked phenotypic heterogeneity. This paper describes the clinical, morphological and laboratory features of an unusual phenotype in a patient harboring the m.8344A>G 'MERRF' mutation. We present the case of a middle-aged woman with distal weakness since childhood who also had ptosis and facial weakness and who developed mid-life respiratory insufficiency necessitating non-invasive nocturnal ventilator support. Neurophysiological and acetylcholine receptor antibody analyses excluded myasthenia gravis whilst molecular genetic testing excluded myotonic dystrophy, prompting a diagnostic needle muscle biopsy. Mitochondrial histochemical abnormalities including subsarcolemmal mitochondrial accumulation (ragged-red fibers) and in excess of 90% COX-deficient fibers, was seen leading to sequencing of the mitochondrial genome in muscle. This identified the m.8344A>G mutation commonly associated with the MERRF phenotype. This case extends the evolving phenotypic spectrum of the m.8344A>G mutation and emphasizes that it may cause indolent distal weakness with respiratory insufficiency, with marked histochemical defects in muscle. Our findings support consideration of screening of this gene in cases of indolent myopathy resembling distal limb-girdle muscular dystrophy in which screening of the common genes prove negative.

  15. Influence of low-power laser radiation on the activity of some membraneous and mitochondrial enzymes of hepatocytes in rats

    Science.gov (United States)

    Cieslar, Grzegorz; Adamek, Mariusz; Sieron, Aleksander; Kaminski, Marcin

    1995-01-01

    It was observed in some experiments that visible laser radiation activates the enzymatic function of mitochondria, while infrared laser radiation affects the enzymatic activity of cellular membranes. The aim of the study was to estimate the activity of some membranous as well as mitochondrial enzymes of hepatocytes in rats irradiated with infrared laser. Experimental material consisted of 38 Wistar rats divided into 2 groups -- a studied group exposed to infrared laser radiation and a control group, in which no irradiation was made. A semiconductive infrared laser (wavelength -- 904 nm, mean power -- 8.9 mW) was used. The clean-shaven skin of the right infracostal region of animals was irradiated 5 minutes daily for 15 consecutive days. After finishing the experiment in the preparations from obtained segments of the left liver lobe, the enzymatic activity of succinate dehydrogenase (SDH, EC 1.3.99.1), lactic dehydrogenase (LDH, EC 1.1.1.27), Mg2+ dependent ATP-ase (ATP-ase Mg2+, EC 3.1.3.2.) and acid phosphatase (AcP, EC 3.6.1.8.) was estimated with the use of histochemical methods. In the case of SDH and LDH the increase of enzymatic activity was observed in all 3 zones of liver cluster, especially in male rats. In the case of ATP-ase Mg2+ and AcP the increase of enzymatic activity in biliary canaliculi of hepatocytes in all zones of the liver cluster was observed. On the basis of the obtained results it was proved that infrared laser radiation activates significantly the enzymatic activity of most of the analyzed enzymes, which means that it affects not only properties of biological membranes but also activates the oxidoreductive processes of organism, as it has been observed for visible laser radiation. On the basis of the spectrum of energetic levels in macromolecules (Jablonski's diagram) the mechanisms of availed results are discussed both for enzymes possessing and not possessing chromatophores.

  16. Aspectos histológicos e histoquímicos da cloaca feminina de Columba livia (Gmelin (Columbidae, Columbiformes Histological and histochemical aspects of female cloaca of Columba livia (Gmelin (Columbidae, Columbiforme

    Directory of Open Access Journals (Sweden)

    Maria Eloíza de Oliveira Teles

    2001-03-01

    Full Text Available The domestic dove Columba livia (Gmelin, 1789 is a species well adapted to Brazil and the study of its reproductive biology is part of a broad research project on birds. This essay describes the morphological aspects of the cloaca of female Columba livia, describing compartments lengthwise starting from the head such as the coprodeo, urodeo and proctodeo limited by mucosal folds. Each compartment of the cloaca presents its own morphological characteristics which differ one from the other in form, height and position of mucosal projections, kinds of tissues, presence and histological aspects of glands, presence of lymphonodus. The rectum opens into the coprodeo, the ureter opens into the oviduct and the cloacal sac opening into the proctodeo. Histochemical studies have shown the presence of glycoproteins in tissue cells and gland cells on the three segments of the cloaca.

  17. Recent trends in nanomaterials immobilised enzymes for biofuel production.

    Science.gov (United States)

    Verma, Madan L; Puri, Munish; Barrow, Colin J

    2016-01-01

    Application of nanomaterials as novel supporting materials for enzyme immobilisation has generated incredible interest in the biotechnology community. These robust nanostructured forms, such as nanoparticles, nanofibres, nanotubes, nanoporous, nanosheets, and nanocomposites, possess a high surface area to volume ratios that can cause a high enzyme loading and facilitate reaction kinetics, thus improving biocatalytic efficiency for industrial applications. In this article, we discuss research opportunities of nanoscale materials in enzyme biotechnology and highlight recent developments in biofuel production using advanced material supports for enzyme immobilisation and stabilisation. Synthesis and functionalisation of nanomaterial forms using different methods are highlighted. Various simple and effective strategies designed to result in a stable, as well as functional protein-nanomaterial conjugates are also discussed. Analytical techniques confirming enzyme loading on nanomaterials and assessing post-immobilisation changes are discussed. The current status of versatile nanomaterial support for biofuel production employing cellulases and lipases is described in details. This report concludes with a discussion on the likely outcome that nanomaterials will become an integral part of sustainable bioenergy production.

  18. Characteristics of enzyme hydrolysis of cellulose under static condition.

    Science.gov (United States)

    Taneda, Daisuke; Ueno, Yoshiki; Ikeo, Makoto; Okino, Shohei

    2012-10-01

    The effect of enzyme loading under static and agitated conditions was investigated. Enzymatic hydrolysis of 10 w/v% de-lignified cellulose slurry such as filter paper, avicel and pulp was conducted under agitated (120 rpm) and static condition, and the enzyme loading ranging from 1.2 to 120 mg-protein/g-dry substrate. Under the agitated condition, the final sugar concentration decreased with the decreasing enzyme loading. Under the static condition, the final sugar concentration was maintained even if the enzyme loading was decreased. The above phenomenon was caused by a rapid precipitation of cellobiohydrolase 2 (CBH2) under the agitated condition, which was not observed under the static condition. The hydrolysis experiments using enzymes containing different ratios of cellobiohydrolase 1 (CBH1) and CBH2 under the static condition suggested that preservation of CBH2 and its synergism with CBH1 is essential for static condition's characteristics, and for efficient hydrolysis of cellulose. Copyright © 2012 Elsevier Ltd. All rights reserved.

  19. Kinetic aspects of human placental alkaline phosphatase enzyme membrane.

    Science.gov (United States)

    Roig, M G; Serrano, M A; Bello, J F; Cachaza, J M; Kennedy, J F

    1991-01-01

    The crosslinking of alkaline phosphatase of human placenta with human serum albumin has been optimized. During the physico-chemical characterization of this immobilized biocatalyst, special attention was paid to attributes such as the irreversibility of the enzyme support bonding, the stability of the catalytic activity, and the effects of pH and temperature on this activity. Regarding stability, patterns of denaturation are proposed, to account for inactivation curves over time and under storage/operation conditions. These patterns, in some cases, indicate the existence of different populations of immobilized enzyme molecules, with a different degree of sensitivity to denaturation. The activity vs pH profiles are clearly modified by the immobilization process. This is because the pH of the free homogeneous solution, measurable with a pH-meter, differs from the real pH of the immediate microenvironment of the immobilized enzyme molecules due to the effects of proton accumulation in the microenvironment (in the reaction catalysed by alkaline phosphatase, protons are produced), to limitations to the free diffusion of H+ and to the possible partition effects of H+ due to polar interactions with residues or molecules of the enzyme membrane. In the experimental working conditions, the apparent optimum temperatures are centered at 40 degrees C, inactivation (thermal denaturation) occurring above this temperature. In the temperature range 10-40 degrees C, the kinetic control over the overall activity of the immobilized enzyme was observed, causing the Arrhenius profiles to be linear.

  20. Effect of photoperiod on gibberellin biosynthetic enzymes in spinach

    Energy Technology Data Exchange (ETDEWEB)

    Gilmour, S.J.; Bleecker, A.B.; Zeevaart, J.A.D.

    1986-04-01

    The photoperiodic control of stem elongation in spinach, a long day (LD) rosette plant, is mediated by gibberellins (GAs). The early 13-hydroxylated GA biosynthetic pathway from GA/sub 12/ to GA/sub 20/ operates in spinach: GA/sub 12/ ..-->.. GA/sub 53/ ..-->.. GA/sub 44/ ..-->.. GA/sub 19/ ..-->.. GA/sub 20/. Two enzymes of this pathway, those converting GA/sub 53/ to GA/sub 44/ (GA/sub 53/ oxidase) and GA/sub 19/ to GA/sub 20/ (GA/sub 19/ oxidase), are regulated by light. The enzyme converting GA/sub 44/ to GA/sub 19/ (GA/sub 44/ oxidase) is not light-regulated. In the light GA/sub 53/ and GA/sub 18/ oxidase activities are increased, therefore causing the GA biosynthetic pathway to be turned on. This leads to the production of an active GA in LD, which causes an increase in stem elongation. Two the enzymes, GA/sub 44/ and GA/sub 53/ oxidases, can be separated from one another by anion exchange HPLC. Estimates of the molecular weights of these two enzymes based on gel filtration HPLC will be reported.

  1. Experimental strategy to discover microbes with gluten-degrading enzyme activities

    Science.gov (United States)

    Helmerhorst, Eva J.; Wei, Guoxian

    2014-06-01

    Gluten proteins contained in the cereals barley, rye and wheat cause an inflammatory disorder called celiac disease in genetically predisposed individuals. Certain immunogenic gluten domains are resistant to degradation by mammalian digestive enzymes. Enzymes with the ability to target such domains are potentially of clinical use. Of particular interest are gluten-degrading enzymes that would be naturally present in the human body, e.g. associated with resident microbial species. This manuscript describes a selective gluten agar approach and four enzyme activity assays, including a gliadin zymogram assay, designed for the selection and discovery of novel gluten-degrading microorganisms from human biological samples. Resident and harmless bacteria and/or their derived enzymes could potentially find novel applications in the treatment of celiac disease, in the form of a probiotic agent or as a dietary enzyme supplement.

  2. THE DEINKING OF NON-IMPACT PRINTED PAPER BY COMBINING ENZYME WITH MECHANICAL ACTION

    Institute of Scientific and Technical Information of China (English)

    Gaosheng Wang; Laisu Xie; Yanquan Long

    2004-01-01

    We studied an enzymatic deinking process of non-impact printed paper which was carried out in batch equipment, similar to Valley beater. The equipment can provide suitable forces to cause toner separation from fiber rather than fiber damage. The factors that influenced the deinking efficiency included beating time, pressure between rotating bars and bottom bars, enzyme properties such as dosage,cellulase activity, temperature, pH, etc. Beating time and enzyme type and enzyme dosage was investigated in detail. The deinked pulp was brighter and cleaner. The drainage property also can be controlled by adjusting enzyme dosage and mechanical action strength. Combining enzyme with mechanical action provides the best deinking effects,not enzyme alone. In the meantime, mechanism of enzymatic deinking was discussed depending on the deinked pulp properties.

  3. THE DEINKING OF NON-IMPACT PRINTED PAPER BY COMBINING ENZYME WITH MECHANICAL ACTION

    Institute of Scientific and Technical Information of China (English)

    GaoshengWang; LaisuXie; YanquanLong

    2004-01-01

    We studied an enzymatic deinking process of non-impact printed paper which was carried out in batch equipment, similar to Valley beater. The equipment can provide suitable forces to cause toner separation from fiber rather than fiber damage. The factors that influenced the deinking efficiency included beating time, pressure between rotating bars and bottom bars, enzyme properties such as dosage, cellulase activity, temperature, pH, etc. Beating time and enzyme type and enzyme dosage was investigated in detail. The deinked pulp was brighter and cleaner. The drainage property also can be controlled by adjusting enzyme dosage and mechanical action strength. Combining enzyme with mechanical action provides the best deinking effects, not enzyme alone. In the meantime, mechanism of enzymatic deinking was discussed depending on the deinked pulp properties.

  4. MORFOLOGICAL AND HISTOCHEMICAL OBSERVATIONS ON THE SALIVARY GLANDS OF THE ALLIGATOR Caiman latirostris OBSERVAÇÕES MORFOLÓGICAS E HISTOQUÍMICAS SOBRE AS GLÂNDULAS SALIVARES DO JACARÉ Caiman latirostris (Crocodilia Reptilia

    Directory of Open Access Journals (Sweden)

    Gilson Herbert de Faria

    2007-09-01

    Full Text Available

    Using histological and histochemical methods for glicoconjugates, the present work on the salivary glands of the alligator Caiman latirostris (Crocodilia Reptilia, was developed. On the bases of the results obtained, the authors believe that those glands are not well developed, appearing as unicellular structures goblet-like cells, in the stratified squamous epithelium of the oral cavity. Scattered glands alveolus without excretory tubules which extend directly with the epitelium were also present. By means of histochemical methods for glicoconjugates, it was possible to detect the glicoprotein and the sialic acid in the glandular formation of these animals.

    Empregando-se métodos histológicos e histoquímicos para glicoconjugados, desenvolveu-se um trabalho sobre glândulas salivares do jacaré Caiman latirostris. Com base nos resultados obtidos, constatou-se que estas glândulas são pouco desenvolvidas, aparecendo como estruturas unicelulares, semelhantes às células caliciformes, incluídas no epitélio estratificado pavimentoso da cavidade oral. Observou-se, ainda, a presença de esparsos alvéolos glandulares desprovidos de ductos excretores que continuam diretamente com o epitélio. Por meio de métodos histoquímicos para glicoconjugados, foi possível a detecção de glicoproteína e de ácido siálico nas formações glandulares desses animais.

  5. Differential responses to acute administration of a new 5-HT7-R agonist as a function of adolescent pre-treatment: phMRI and immuno-histochemical study.

    Science.gov (United States)

    Altabella, Luisa; Sbriccoli, Marco; Zoratto, Francesca; Poleggi, Anna; Vinci, Ramona; Lacivita, Enza; Leopoldo, Marcello; Laviola, Giovanni; Cardone, Franco; Canese, Rossella; Adriani, Walter

    2014-01-01

    LP-211 is a new, selective agonist of serotonin (5-hydroxytryptamine, 5-HT) receptor 7 (5-HT7-R), which is part of a neuro-transmission system with a proposed role in neural plasticity and in mood, cognitive and sleep regulation. Adolescent subchronic LP-211 treatment produces some persisting changes in rats' forebrain structural and functional parameters. Here, using pharmacological MRI (phMRI), we investigated the effect of acute administration with LP-211 (10 mg/kg i.p.), or vehicle, to adult rats previously exposed to the same drug (0.25 mg/kg/day for 5 days), or vehicle, during adolescence (44-48 post-natal days); histology and immuno-histochemistry were performed ex vivo to evaluate neuro-anatomical and physiological long-term adaptation to pharmacological pre-treatment. The phMRI signal reveals forebrain areas (i.e., hippocampus, orbital prefrontal cortex), activated in response to LP-211 challenge independently of adolescent pre-treatment. In septum and nucleus accumbens, sensitized activation was found in adolescent pre-treated rats but not in vehicle-exposed controls. Immuno-histochemical analyses showed marked differences in septum as long-term consequence of the adolescent pre-treatment: increased level of 5-HT7-R, increased number of 5-HT7-R positive cells, and enhanced astrocyte activation. For nucleus accumbens, immuno-histochemical analyses did not reveal any difference between adolescent pre-treated rats and vehicle-exposed controls. In conclusion, subchronic LP-211 administration during adolescence is able to induce persistent physiological changes in the septal 5-HT7-R expression and astrocyte response that can still be observed in adulthood. Data shed new insights into roles of 5-HT7-R for normal and pathologic behavioral regulations.

  6. Differential responses to acute administration of a new 5-HT7-R agonist as a function of adolescent pre-treatment: phMRI and immuno-histochemical study

    Directory of Open Access Journals (Sweden)

    Luisa eAltabella

    2014-12-01

    Full Text Available LP-211 is a new, selective agonist of serotonin (5-hydroxytryptamine, 5-HT receptor 7 (5-HT7-R, which is part of a neuro-transmission system with a proposed role in neural plasticity and in mood, cognitive or sleep regulation. Adolescent subchronic LP-211 treatment produces some persisting changes in rats’ forebrain structural and functional parameters. Here, using pharmacological MRI (phMRI, we investigated the effect of acute administration with LP-211 (10 mg/kg i.p., or vehicle, to adult rats previously exposed to the same drug (0.25 mg/kg/day for 5 days, or vehicle, during adolescence (44 to 48 post-natal days; histology and immuno-histochemistry were performed ex vivo to evaluate neuro-anatomical and physiological long-term adaptation to pharmacological pre-treatment. The phMRI signal reveals forebrain areas (i.e. hippocampus, orbital prefrontal cortex, activated in response to LP-211 challenge independently of adolescent pre-treatment. In septum and nucleus accumbens, sensitized activation was found in adolescent pre-treated rats but not in vehicle-exposed controls. Immuno-histochemical analyses showed marked differences in septum as long-term consequence of the adolescent pre-treatment: increased level of 5-HT7-R, increased number of 5-HT7-R positive cells, and enhanced astrocyte activation. For nucleus accumbens, immuno-histochemical analyses did not reveal any difference between adolescent pre-treated rats and vehicle-exposed controls. In conclusion, subchronic LP-211 administration during adolescence is able to induce persistent physiological changes in the septal 5-HT7-R expression and astrocyte response that can still be observed in adulthood. Data shed new insights into roles of 5-HT7-R for normal and pathologic behavioral regulations.

  7. Castor Oil Transesterification Catalysed by Liquid Enzymes

    DEFF Research Database (Denmark)

    Andrade, Thalles; Errico, Massimiliano; Christensen, Knud Villy

    2017-01-01

    In the present work, biodiesel production by reaction of non-edible castor oil with methanol under enzymatic catalysis is investigated. Two liquid enzymes were tested: Eversa Transform and Resinase HT. Reactions were performed at 35 °C and with a molar ratio of methanol to oil of 6:1. The reaction...... time was 8 hours. Stepwise addition of methanol was necessary to avoid enzyme inhibition by methanol. In order to minimize the enzyme costs, the influence of enzyme activity loss during reuse of both enzymes was evaluated under two distinct conditions. In the former, the enzymes were recovered...... and fully reused; in the latter, a mixture of 50 % reused and 50 % fresh enzymes was tested. In the case of total reuse after three cycles, both enzymes achieved only low conversions. The biodiesel content in the oil-phase using Eversa Transform was 94.21 % for the first cycle, 68.39 % in the second, and 33...

  8. Enzymic synthesis of labelled chiral substances.

    Science.gov (United States)

    Battersby, A R

    1985-01-01

    The enzymic synthesis of chiral substances in which one hydrogen atom of a methylene group has been replaced by deuterium or tritium is illustrated. Such labelled products can be used to determine the stereochemistry of other enzyme-catalysed reactions.

  9. Activation of thiamin diphosphate in enzymes.

    Science.gov (United States)

    Hübner, G; Tittmann, K; Killenberg-Jabs, M; Schäffner, J; Spinka, M; Neef, H; Kern, D; Kern, G; Schneider, G; Wikner, C; Ghisla, S

    1998-06-29

    Activation of the coenzyme ThDP was studied by measuring the kinetics of deprotonation at the C2 carbon of thiamin diphosphate in the enzymes pyruvate decarboxylase, transketolase, pyruvate dehydrogenase complex, pyruvate oxidase, in site-specific mutant enzymes and in enzyme complexes containing coenzyme analogues by proton/deuterium exchange detected by 1H-NMR spectroscopy. The respective deprotonation rate constant is above the catalytic constant in all enzymes investigated. The fast deprotonation requires the presence of an activator in pyruvate decarboxylase from yeast, showing the allosteric regulation of this enzyme to be accomplished by an increase in the C2-H dissociation rate of the enzyme-bound thiamin diphosphate. The data of the thiamin diphosphate analogues and of the mutant enzymes show the N1' atom and the 4'-NH2 group to be essential for the activation of the coenzyme and a conserved glutamate involved in the proton abstraction mechanism of the enzyme-bound thiamin diphosphate.

  10. Editorial: Special Issue — Enzyme Immobilization

    Directory of Open Access Journals (Sweden)

    Roberto Fernandez-Lafuente

    2014-12-01

    Full Text Available Immobilization of enzymes and proteins is a requirement for many industrial enzyme applications, as this facilitates enzyme recovery and reuse. Bearing in mind this necessity, the coupling of immobilization to the improvement of other enzyme features has been pursued by many researchers, and nowadays immobilization is recognized as a tool to improve not only stability, but also enzyme selectivity, specificity, resistance to inhibition or chemical modifiers, etc. To achieve these overall improvements of enzymes’ properties via immobilization, it is necessary to both develop new immobilization systems suitable for these purposes, and to achieve a deeper knowledge of the mechanisms of interaction between enzymes and activated solids. That way, immobilization of enzymes, far being an old-fashioned methodology to just reuse these expensive biocatalysts, is a tool of continuous interest that requires a continuous effort to be exploited in all its potential. This special issue collects 23 papers reporting advances in the field of immobilization of enzymes.[...

  11. A DNA tweezer-actuated enzyme nanoreactor.

    Science.gov (United States)

    Liu, Minghui; Fu, Jinglin; Hejesen, Christian; Yang, Yuhe; Woodbury, Neal W; Gothelf, Kurt; Liu, Yan; Yan, Hao

    2013-01-01

    The functions of regulatory enzymes are essential to modulating cellular pathways. Here we report a tweezer-like DNA nanodevice to actuate the activity of an enzyme/cofactor pair. A dehydrogenase and NAD(+) cofactor are attached to different arms of the DNA tweezer structure and actuation of enzymatic function is achieved by switching the tweezers between open and closed states. The enzyme/cofactor pair is spatially separated in the open state with inhibited enzyme function, whereas in the closed state, enzyme is activated by the close proximity of the two molecules. The conformational state of the DNA tweezer is controlled by the addition of specific oligonucleotides that serve as the thermodynamic driver (fuel) to trigger the change. Using this approach, several cycles of externally controlled enzyme inhibition and activation are successfully demonstrated. This principle of responsive enzyme nanodevices may be used to regulate other types of enzymes and to introduce feedback or feed-forward control loops.

  12. Extracellular enzyme kinetics scale with resource availability

    Science.gov (United States)

    Microbial community metabolism relies on external digestion, mediated by extracellular enzymes that break down complex organic matter into molecules small enough for cells to assimilate. We analyzed the kinetics of 40 extracellular enzymes that mediate the degradation and assimi...

  13. What Causes Bronchitis?

    Science.gov (United States)

    ... page from the NHLBI on Twitter. What Causes Bronchitis? Acute Bronchitis Infections or lung irritants cause acute bronchitis. The ... fire, also may lead to acute bronchitis. Chronic Bronchitis Repeatedly breathing in fumes that irritate and damage ...

  14. What Causes Lactose Intolerance?

    Science.gov (United States)

    ... Information Clinical Trials Resources and Publications What causes lactose intolerance? Skip sharing on social media links Share ... lactase in the body is the cause of lactose intolerance. The names for the three types of ...

  15. Causes of Pediatric Cardiomyopathy

    Science.gov (United States)

    Search ABOUT THE DISEASE CAUSES Although pediatric cardiomyopathy is one of the leading causes of cardiac death in children, an explanation for why it occurs remains unknown. Most cases are familial ...

  16. What Causes Cardiomyopathy?

    Science.gov (United States)

    ... page from the NHLBI on Twitter. What Causes Cardiomyopathy? Cardiomyopathy can be acquired or inherited. “Acquired” means ... case when the disease occurs in children. Hypertrophic Cardiomyopathy Hypertrophic cardiomyopathy usually is inherited. It’s caused by ...

  17. What Causes Menstrual Irregularities?

    Science.gov (United States)

    ... Publications What causes menstrual irregularities? Skip sharing on social media links Share this: Page Content Menstrual irregularities can be caused by a variety of conditions, including pregnancy, hormonal imbalances, infections, malignancies, diseases, trauma, and certain ...

  18. What Causes Atherosclerosis?

    Science.gov (United States)

    ... page from the NHLBI on Twitter. What Causes Atherosclerosis? The exact cause of atherosclerosis isn't known. ... Rate This Content: NEXT >> Featured Video What is atherosclerosis? 05/22/2014 Describes how the build-up ...

  19. What Causes Anemia?

    Science.gov (United States)

    ... page from the NHLBI on Twitter. What Causes Anemia? The three main causes of anemia are: Blood ... the blood and can lead to anemia. Aplastic Anemia Some infants are born without the ability to ...

  20. What Causes Cystic Fibrosis?

    Science.gov (United States)

    ... page from the NHLBI on Twitter. What Causes Cystic Fibrosis? A defect in the CFTR gene causes cystic ... in the severity of the disease. How Is Cystic Fibrosis Inherited? Every person inherits two CFTR genes—one ...

  1. What Causes Sarcoidosis?

    Science.gov (United States)

    ... page from the NHLBI on Twitter. What Causes Sarcoidosis? The cause of sarcoidosis isn't known. More ... Content: NEXT >> Featured Video Living With and Managing Sarcoidosis 05/18/2011 This video—presented by the ...

  2. What Causes Down Syndrome?

    Science.gov (United States)

    ... Information Clinical Trials Resources and Publications What causes Down syndrome? Skip sharing on social media links Share this: ... Down Syndrome Registry​ . Chromosomal Changes That Can Cause Down Syndrome Research shows that three types of chromosomal changes ...

  3. Causes of High Cholesterol

    Science.gov (United States)

    ... Venous Thromboembolism Aortic Aneurysm More Causes of High Cholesterol Updated:Jul 5,2017 If you have high ... and procedures related to heart disease and stroke. Cholesterol • Home • About Cholesterol • HDL, LDL, and Triglycerides • Causes ...

  4. Incidence Causes and Outcome

    African Journals Online (AJOL)

    GB

    causes and outcome of obstructed labor in Jimma University Specialized Hospital. METHODS: ... Obstructed labor is an important cause of ... and outcome on the mother and baby. Data .... to this hospital after onset of labor as compared to the.

  5. Biomedical Applications of Enzymes From Marine Actinobacteria.

    Science.gov (United States)

    Kamala, K; Sivaperumal, P

    Marine microbial enzyme technologies have progressed significantly in the last few decades for different applications. Among the various microorganisms, marine actinobacterial enzymes have significant active properties, which could allow them to be biocatalysts with tremendous bioactive metabolites. Moreover, marine actinobacteria have been considered as biofactories, since their enzymes fulfill biomedical and industrial needs. In this chapter, the marine actinobacteria and their enzymes' uses in biological activities and biomedical applications are described. © 2017 Elsevier Inc. All rights reserved.

  6. What Causes Heart Disease?

    Science.gov (United States)

    ... page from the NHLBI on Twitter. What Causes Heart Disease? Research suggests that coronary heart disease (CHD) begins with damage to the lining and ... causing coronary microvascular disease (MVD). Coronary MVD is heart disease that affects the heart's tiny arteries. The cause ...

  7. [Liver damage caused by drugs].

    Science.gov (United States)

    Strohmeyer, G; Weik, C

    1999-05-01

    The liver has a central role in the metabolism of many drugs, since this organ is the main site of biotransformation of endo- and xenobiotics. Water-soluble drugs have a small volume of distribution and can be eliminated unchanged in the urine. By contrast, lipid-soluble drugs have a larger volume of distribution and require conversion to water-soluble metabolites for their elimination in urine or bile. The liver with its specific receptors, transporters and enzymes is responsible for the uptake, transformation and excretion of the lipophilic drugs. While most of the drugs are transformed into stable metabolites, other drugs form reactive, potentially toxic, metabolites producing liver cell damage. Liver injury caused by drugs may mimic almost any kind of liver disease. Clinical findings are gastrointestinal symptoms with nausea, vomiting and abdominal pain, cholestatic liver injury with jaundice and pruritus of severe inflammatory and cirrhotic liver damage with signs of liver failure, encephalopathy and cerebral edema. The morphological changes vary from hepatitis, cholestasis, fatty liver, granulomatous hepatitis, peri-/portal inflammation, to fibrosis with cirrhotic alterations and vascular lesions and tumors. The most commonly used drugs causing severe liver injury are discussed in detail. These are anabolics, oral contraceptives, antituberculous and antifungal agents, nonsteroidal anti-inflammatory drugs, ring substituted amphetamins ("designer drugs"), antiarrhythmics and antibiotics.

  8. Cellulolytic enzyme compositions and uses thereof

    Energy Technology Data Exchange (ETDEWEB)

    Iyer, Prashant; Gaspar, Armindo Ribiero; Croonenberghs, James; Binder, Thomas P.

    2017-07-25

    The present invention relates enzyme composition comprising a cellulolytic preparation and an acetylxylan esterase (AXE); and the used of cellulolytic enzyme compositions for hydrolyzing acetylated cellulosic material. Finally the invention also relates to processes of producing fermentation products from acetylated cellulosic materials using a cellulolytic enzyme composition of the invention.

  9. Determining Enzyme Activity by Radial Diffusion

    Science.gov (United States)

    Davis, Bill D.

    1977-01-01

    Discusses advantages of radial diffusion assay in determining presence of enzyme and/or rough approximation of amount of enzyme activities. Procedures are included for the preparation of starch-agar plates, and the application and determination of enzyme. Techniques using plant materials (homogenates, tissues, ungerminated embryos, and seedlings)…

  10. Determining Enzyme Activity by Radial Diffusion

    Science.gov (United States)

    Davis, Bill D.

    1977-01-01

    Discusses advantages of radial diffusion assay in determining presence of enzyme and/or rough approximation of amount of enzyme activities. Procedures are included for the preparation of starch-agar plates, and the application and determination of enzyme. Techniques using plant materials (homogenates, tissues, ungerminated embryos, and seedlings)…

  11. 21 CFR 864.4400 - Enzyme preparations.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Enzyme preparations. 864.4400 Section 864.4400...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Specimen Preparation Reagents § 864.4400 Enzyme preparations. (a) Identification. Enzyme preparations are products that are used in the...

  12. Consumer attitudes to enzymes in food production

    DEFF Research Database (Denmark)

    Søndergaard, Helle Alsted; Grunert, Klaus G.; Scholderer, Joachim

    2005-01-01

    The use of enzymes in food production has potential benefits for both food manufacturers and consumers. A central question is how consumers react to new ways of producing foods with enzymes. This study investigates the formation of consumer attitudes to different enzyme production methods in three...

  13. Curious cases of the enzymes

    OpenAIRE

    Ulusu Nuriye Nuray

    2015-01-01

    J Med Biochem 2015; 34 (3) DOI: 10.2478/jomb-2014-0045 UDK 577. 1 : 61 ISSN 1452-8258 J Med Biochem 34: 271–281, 2015 Review article Pregledni ~lanak CURIOUS CASES OF THE ENZYMES NEOBI^NA ISTORIJA ENZIMA Nuriye Nuray Ulusu Koç University, School of Medicine, Sariyer-Istanbul, Turkey Address for correspondence: N. Nuray Ulusu, PhD Koç University School of Medicine Professor of Biochemistry Rumelifeneri Yolu Sarıyer-Istanbul – Turkey Phone: +90 (212)...

  14. ACTIVITY OF SUPEROXIDE DISMUTASE ENZYME IN YEAST SACCHAROMYCES CEREVISIAE

    OpenAIRE

    2014-01-01

    Reactive oxygen species (ROS) with reactive nitrogen species (RNS) are known to play dual role in biological systems, they can be harmful or beneficial to living systems. ROS can be important mediators of damage to cell structures, including proteins, lipids and nucleic acids termed as oxidative stress. The antioxidant enzymes protect the organism against the oxidative damage caused by active oxygen forms. The role of superoxide dismutase (SOD) is to accelerate the dismutation of the toxic su...

  15. Inhibition of Aminoglycoside Acetyltransferase Resistance Enzymes by Metal Salts

    OpenAIRE

    2015-01-01

    Aminoglycosides (AGs) are clinically relevant antibiotics used to treat infections caused by both Gram-negative and Gram-positive bacteria, as well as Mycobacteria. As with all current antibacterial agents, resistance to AGs is an increasing problem. The most common mechanism of resistance to AGs is the presence of AG-modifying enzymes (AMEs) in bacterial cells, with AG acetyltransferases (AACs) being the most prevalent. Recently, it was discovered that Zn2+ metal ions displayed an inhibitory...

  16. Immunohistochemical localization of the antioxidant enzymes biliverdin reductase and heme oxygenase-2 in human and pig gastric fundus.

    Science.gov (United States)

    Colpaert, Erwin E; Timmermans, Jean Pierre; Lefebvre, Romain A

    2002-04-01

    The intrinsic antioxidant capacities of the bile pigments biliverdin and bilirubin are increasingly recognized since both heme degradation products can exert beneficial cytoprotective effects due to their scavenging of oxygen free radicals and interaction with antioxidant vitamins. Several studies have been published on the localization of the carbon monoxide producing enzyme heme oxygenase-2 (HO-2), which concomitantly generates biliverdin; histochemical data on the distribution of biliverdin reductase (BVR), converting biliverdin to bilirubin, are still very scarce in large mammals including humans. The present study revealed by means of immunohistochemistry the presence of BVR and HO-2 in mucosal epithelial cells and in the endothelium of intramural vessels of both human and porcine gastric fundus. In addition, co-labeling with the specific neural marker protein-gene product 9.5 (PGP 9.5) demonstrated that both BVR and HO-2 were present in all intrinsic nerve cell bodies of both submucous and myenteric plexuses, while double labeling with c-Kit antibody confirmed their presence in intramuscular interstitial cells of Cajal (ICC). Our results substantiate the hypothesis that BVR, through the production of the potent antioxidant bilirubin, might be an essential component of normal physiologic gastrointestinal defense in man and pig.

  17. Enzyme surface rigidity tunes the temperature dependence of catalytic rates.

    Science.gov (United States)

    Isaksen, Geir Villy; Åqvist, Johan; Brandsdal, Bjørn Olav

    2016-07-12

    The structural origin of enzyme adaptation to low temperature, allowing efficient catalysis of chemical reactions even near the freezing point of water, remains a fundamental puzzle in biocatalysis. A remarkable universal fingerprint shared by all cold-active enzymes is a reduction of the activation enthalpy accompanied by a more negative entropy, which alleviates the exponential decrease in chemical reaction rates caused by lowering of the temperature. Herein, we explore the role of protein surface mobility in determining this enthalpy-entropy balance. The effects of modifying surface rigidity in cold- and warm-active trypsins are demonstrated here by calculation of high-precision Arrhenius plots and thermodynamic activation parameters for the peptide hydrolysis reaction, using extensive computer simulations. The protein surface flexibility is systematically varied by applying positional restraints, causing the remarkable effect of turning the cold-active trypsin into a variant with mesophilic characteristics without changing the amino acid sequence. Furthermore, we show that just restraining a key surface loop causes the same effect as a point mutation in that loop between the cold- and warm-active trypsin. Importantly, changes in the activation enthalpy-entropy balance of up to 10 kcal/mol are almost perfectly balanced at room temperature, whereas they yield significantly higher rates at low temperatures for the cold-adapted enzyme.

  18. Consumer attitudes to enzymes in food production

    DEFF Research Database (Denmark)

    Søndergaard, Helle Alsted; Grunert, Klaus G.; Scholderer, Joachim

    2005-01-01

    The use of enzymes in food production has potential benefits for both food manufacturers and consumers. A central question is how consumers react to new ways of producing foods with enzymes. This study investigates the formation of consumer attitudes to different enzyme production methods in three...... European countries. Results show that consumers are most positive towards non-GM enzyme production methods. The enzyme production method is by far the most important factor for the formation of buying intentions compared to price and benefits. Results also show that environmental concern and attitudes...

  19. Enzyme Analysis to Determine Glucose Content

    Science.gov (United States)

    Carpenter, Charles; Ward, Robert E.

    Enzyme analysis is used for many purposes in food science and technology. Enzyme activity is used to indicate adequate processing, to assess enzyme preparations, and to measure constituents of foods that are enzyme substrates. In this experiment, the glucose content of corn syrup solids is determined using the enzymes, glucose oxidase and peroxidase. Glucose oxidase catalyzes the oxidation of glucose to form hydrogen peroxide (H2O2), which then reacts with a dye in the presence of peroxidase to give a stable colored product.

  20. Direct Electron Transfer of Enzymes in a Biologically Assembled Conductive Nanomesh Enzyme Platform.

    Science.gov (United States)

    Lee, Seung-Woo; Lee, Ki-Young; Song, Yong-Won; Choi, Won Kook; Chang, Joonyeon; Yi, Hyunjung

    2016-02-24

    Nondestructive assembly of a nanostructured enzyme platform is developed in combination of the specific biomolecular attraction and electrostatic coupling for highly efficient direct electron transfer (DET) of enzymes with unprecedented applicability and versatility. The biologically assembled conductive nanomesh enzyme platform enables DET-based flexible integrated biosensors and DET of eight different enzyme with various catalytic activities.

  1. Causes and effects.

    Science.gov (United States)

    Cone, Carol L; Feldman, Mark A; DaSilva, Alison T

    2003-07-01

    Most companies make charitable donations, but few approach their contributions with an eye toward enhancing their brands. Those that do take such an approach commit talent and know-how, not just dollars, to a pressing but carefully chosen social need and then tell the world about the cause and their service to it. Through the association, both the business and the cause benefit in ways they could not otherwise. Organizations such as Avon, ConAgra Foods, and Chevrolet have recognized that a sustained cause-branding program can improve their reputations, boost their employees' morale, strengthen relations with business partners, and drive sales. And the targeted causes receive far more money than they could have from direct corporate gifts alone. The authors examine these best practices and offer four principles for building successful cause-branding programs. First, they say, a company should select a cause that advances its corporate goals. That is, unless the competitive logic for supporting the cause is clear, a company shouldn't even consider putting its finite resources behind it. Second, a business should commit to a cause before picking its charitable partners. Otherwise, a cause-branding program may become too dependent on its partners. Third, a company should put all its assets to work, especially its employees. It should leverage the professional skills of its workers as well as its other assets such as distribution networks. And fourth, a company should promote its philanthropic initiatives through every possible channel. In addition to using the media, it should communicate its efforts through the Web, annual reports, direct mail, and so on. Cause branding is a way to turn the obligations of corporate citizenship into a valuable asset. When the cause is well chosen, the commitment genuine, and the program well executed, the cause helps the company, and the company helps the cause.

  2. Enzyme extraction by ultrasound from sludge flocs

    Institute of Scientific and Technical Information of China (English)

    YU Guanghui; HE Pinjing; SHAO Liming; ZHU Yishu

    2009-01-01

    Enzymes play essential roles in the biological processes of sludge treatment. In this article, the ultrasound method to extract enzymes from sludge flocs was presented. Results showed that using ultrasound method at 20 kHz could extract more types of enzymes than that ultrasound at 40 kHz and ethylenediamine tetraacetic acid (EDTA) methods. The optimum parameters of ultrasound extraction at 20 kHz were duration of 10 min and power of 480 W. Under the condition, ultrasound could break the cells and extract both the extracellular and intercellular enzymes. Ultrasound power was apparently more susceptive to enzyme extraction than duration, suggesting that the control of power during ultrasound extraction was more important than that of duration. The Pearson correlation analysis between enzyme activities and cation contents revealed that the different types of enzymes had distinct cation binding characteristics.

  3. DNA-Based Enzyme Reactors and Systems

    Directory of Open Access Journals (Sweden)

    Veikko Linko

    2016-07-01

    Full Text Available During recent years, the possibility to create custom biocompatible nanoshapes using DNA as a building material has rapidly emerged. Further, these rationally designed DNA structures could be exploited in positioning pivotal molecules, such as enzymes, with nanometer-level precision. This feature could be used in the fabrication of artificial biochemical machinery that is able to mimic the complex reactions found in living cells. Currently, DNA-enzyme hybrids can be used to control (multi-enzyme cascade reactions and to regulate the enzyme functions and the reaction pathways. Moreover, sophisticated DNA structures can be utilized in encapsulating active enzymes and delivering the molecular cargo into cells. In this review, we focus on the latest enzyme systems based on novel DNA nanostructures: enzyme reactors, regulatory devices and carriers that can find uses in various biotechnological and nanomedical applications.

  4. CELLULOSE DEGRADATION BY OXIDATIVE ENZYMES

    Directory of Open Access Journals (Sweden)

    Maria Dimarogona

    2012-09-01

    Full Text Available Enzymatic degradation of plant biomass has attracted intensive research interest for the production of economically viable biofuels. Here we present an overview of the recent findings on biocatalysts implicated in the oxidative cleavage of cellulose, including polysaccharide monooxygenases (PMOs or LPMOs which stands for lytic PMOs, cellobiose dehydrogenases (CDHs and members of carbohydrate-binding module family 33 (CBM33. PMOs, a novel class of enzymes previously termed GH61s, boost the efficiency of common cellulases resulting in increased hydrolysis yields while lowering the protein loading needed. They act on the crystalline part of cellulose by generating oxidized and non-oxidized chain ends. An external electron donor is required for boosting the activity of PMOs. We discuss recent findings concerning their mechanism of action and identify issues and questions to be addressed in the future.

  5. Chaos in an enzyme reaction.

    Science.gov (United States)

    Olsen, L F; Degn, H

    1977-05-12

    Dynamic systems are usually thought to have either monotonic or periodic behaviour. Although the possibility of other types of behaviour has been recognised for many years, the existence of non-monotonic, non-periodic behaviour in dynamic systems has been firmly established only recently. It is termed chaotic behaviour. A review on the rapidly expanding literature on chaos in discrete model systems described by difference equations has been published by May. Rössler, on the other hand, has discussed a few published works on systems of differential equations with chaotic solutions, and he has proposed a three-component chemical model system which he argues has chaotic solutions [figure see text]. The argument is based on a theorem by Li and Yorke. Here we report the finding of chaotic behaviour as an experimental result in an enzyme system (peroxidase). Like Rössler we base our identification of chaos on the theorem by Li and Yorke.

  6. Encapsulation of Enzymes and Peptides

    Science.gov (United States)

    Meesters, Gabrie M. H.

    A large part of formulated peptides and proteins, e.g., enzymes used as food ingredients, are formulated in a liquid form. Often, they are dissolved in water to which glycerol or sorbitol is added to reduce the water activity of the liquid, thus reducing the change of microbial growth. Still, there are reasons to formulate them in a solid form. Often, these reasons are stability, since a dry formulation is often much better than liquid formulations, and less transportation cost, since less mass is transported if one gets rid of the liquid; however, most of the times, the reason is that the product is mixed with a solid powder. Here, a liquid addition would lead to lump formation.

  7. Cellulose degradation by oxidative enzymes

    Directory of Open Access Journals (Sweden)

    Maria Dimarogona

    2012-09-01

    Full Text Available Enzymatic degradation of plant biomass has attracted intensive research interest for the production of economically viable biofuels. Here we present an overview of the recent findings on biocatalysts implicated in the oxidative cleavage of cellulose, including polysaccharide monooxygenases (PMOs or LPMOs which stands for lytic PMOs, cellobiose dehydrogenases (CDHs and members of carbohydrate-binding module family 33 (CBM33. PMOs, a novel class of enzymes previously termed GH61s, boost the efficiency of common cellulases resulting in increased hydrolysis yields while lowering the protein loading needed. They act on the crystalline part of cellulose by generating oxidized and non-oxidized chain ends. An external electron donor is required for boosting the activity of PMOs. We discuss recent findings concerning their mechanism of action and identify issues and questions to be addressed in the future.

  8. Ethanologenic Enzymes of Zymomonas mobilis

    Energy Technology Data Exchange (ETDEWEB)

    Ingram, Lonnie O' Neal

    1999-03-01

    Zymomonas mobilis is a unique microorganism in being both obligately fermentative and utilizing a Entner-Doudoroff pathway for glycolysis. Glycolytic flux in this organism is readily measured as evolved carbon dioxide, ethanol, or glucose consumed and exceeds 1 {micro}mole glucose/min per mg cell protein. To support this rapid glycolysis, approximately 50% of cytoplasmic protein is devoted to the 13 glycolytic and fermentative enzymes which constitute this central catabolic pathway. Only 1 ATP (net) is produced from each glucose metabolized. During the past grant period, we have completed the characterization of 11 of the 13 glycolytic genes from Z. mobilis together with complementary but separate DOE-fimded research by a former post-dot and collaborator, Dr. Tyrrell Conway. Research funded in my lab by DOE, Division of Energy Biosciences can be divided into three sections: A. Fundamental studies; B. Applied studies and utility; and C. Miscellaneous investigations.

  9. CAUSES OF OCCUPATIONAL INJURIES

    NARCIS (Netherlands)

    KINGMA, J

    1994-01-01

    The causes of occupational injuries (N = 2,365) were investigated. Accidents with machinery and hand tools were the two main causes (49.9%). 89% of the patients with occupational injuries were male. The highest risk group were in the age category of 19 years or less (51.9%). This age group also show

  10. Empyema caused by trichomonas.

    Science.gov (United States)

    Lewis, Kevin L; Doherty, Dennis E; Ribes, Julie; Seabolt, John P; Bensadoun, Eric S

    2003-01-01

    Empyema is one of the potential complications of lower respiratory tract infections. Very rarely, in predisposed individuals, empyema can be caused by Trichomonas species, of which Trichomonas tenax appears to be the most common cause. Here, we present a case of trichomonal empyema in a 56-year-old man and review the available literature of this rare occurrence.

  11. Causes of Child Abuse

    OpenAIRE

    Deveci,S.Erhan; Açık, Yasemin

    2014-01-01

    Child abuse is an important public health problem that is present almost in every society and environment at different level and intensities. For implementation of child abuse protection measures it is necessary to investigate its causes. In this review, causes of child abuse was attempted to investigate with respects to the society and institution, family and individual and child related factors.

  12. Causes of Paralysis

    Science.gov (United States)

    ... is caused by a virus that attacks the nerves which control motor function. > Spina bifida A neural tube defect that causes incomplete closure in the spinal column. > Spinal cord injury Involves damage to the nerves within the bony protection of the spinal canal. > ...

  13. Do Allergies Cause Asthma?

    Science.gov (United States)

    ... Happens in the Operating Room? Do Allergies Cause Asthma? KidsHealth > For Kids > Do Allergies Cause Asthma? A A A en español ¿Las alergias provocan ... kinds of allergies are more likely to have asthma. Do you have allergies that affect your nose ...

  14. What Causes Bad Breath?

    Science.gov (United States)

    ... A Week of Healthy Breakfasts Shyness What Causes Bad Breath? KidsHealth > For Teens > What Causes Bad Breath? A A A en español ¿Qué es lo que provoca el mal aliento? Bad breath, or halitosis , can be a major problem, ...

  15. What Causes Raynaud's?

    Science.gov (United States)

    ... primary and secondary. In primary Raynaud’s (also called Raynaud’s disease), the cause isn't known. Primary Raynaud's is ... examples of diseases and conditions that can cause Raynaud's include: Rheumatoid (RU-ma-toyd) ... (SHOW-gren's) syndrome, dermatomyositis (DER-ma-to-mi-o-SI-tis), ...

  16. Causes of Diabetes

    Science.gov (United States)

    ... iron can build up in and damage the pancreas and other organs. Hormonal diseases Some hormonal diseases cause the body to produce too much of certain hormones, which sometimes cause insulin resistance and diabetes. ... of the pancreas Pancreatitis , pancreatic cancer, and trauma can all harm ...

  17. Inhibition properties of propolis extracts to some clinically important enzymes.

    Science.gov (United States)

    Baltas, Nimet; Yildiz, Oktay; Kolayli, Sevgi

    2016-01-01

    The present study was conducted to envisage inhibition effects of propolis on the crucial enzymes, urease, xanthine oxidase (XO) and acetylcholinesterase (AChE). Some of the antioxidant properties of the propolis samples were determined using the total phenolic content (TPE) and total flavonoids in the eight different ethanolic propolis extracts (EPE) samples. Inhibition values of the enzymes were expressed as inhibition concentration (IC50; mg/mL or μg/mL) causing 50% inhibition of the enzymes with donepezil, acetohydroxamic acid and allopurinol as reference inhibitors. All the propolis extracts exhibited variable inhibition effects on these enzymes, but the higher the phenolic contents the lower the inhibitions values (IC50 = 0.074 to 1.560 mg/mL). IC50 values of the P5 propolis sample having the highest TPE, obtained from Zonguldak, for AChE, urease and XO were 0.081 ± 0.009, 0.080 ± 0.006 and 0.074 ± 0.011 μg/mL, respectively. The EPE proved to be a good source of inhibitor agents that can be used as natural inhibitors to serve human health.

  18. Characterization of the Entamoeba histolytica ornithine decarboxylase-like enzyme.

    Directory of Open Access Journals (Sweden)

    Anupam Jhingran

    Full Text Available BACKGROUND: The polyamines putrescine, spermidine, and spermine are organic cations that are required for cell growth and differentiation. Ornithine decarboxylase (ODC, the first and rate-limiting enzyme in the polyamine biosynthetic pathway, is a highly regulated enzyme. METHODOLOGY AND RESULTS: To use this enzyme as a potential drug target, the gene encoding putative ornithine decarboxylase (ODC-like sequence was cloned from Entamoeba histolytica, a protozoan parasite causing amoebiasis. DNA sequence analysis revealed an open reading frame (ORF of approximately 1,242 bp encoding a putative protein of 413 amino acids with a calculated molecular mass of 46 kDa and a predicted isoelectric point of 5.61. The E. histolytica putative ODC-like sequence has 33% sequence identity with human ODC and 36% identity with the Datura stramonium ODC. The ORF is a single-copy gene located on a 1.9-Mb chromosome. The recombinant putative ODC protein (48 kDa from E. histolytica was heterologously expressed in Escherichia coli. Antiserum against recombinant putative ODC protein detected a band of anticipated size approximately 46 kDa in E. histolytica whole-cell lysate. Difluoromethylornithine (DFMO, an enzyme-activated irreversible inhibitor of ODC, had no effect on the recombinant putative ODC from E. histolytica. Comparative modeling of the three-dimensional structure of E. histolytica putative ODC shows that the putative binding site for DFMO is disrupted by the substitution of three amino acids-aspartate-332, aspartate-361, and tyrosine-323-by histidine-296, phenylalanine-305, and asparagine-334, through which this inhibitor interacts with the protein. Amino acid changes in the pocket of the E. histolytica enzyme resulted in low substrate specificity for ornithine. It is possible that the enzyme has evolved a novel substrate specificity. CONCLUSION: To our knowledge this is the first report on the molecular characterization of putative ODC-like sequence from

  19. Enzyme-guided DNA Sewing Architecture.

    Science.gov (United States)

    Song, In Hyun; Shin, Seung Won; Park, Kyung Soo; Lansac, Yves; Jang, Yun Hee; Um, Soong Ho

    2015-12-04

    With the advent of nanotechnology, a variety of nanoarchitectures with varied physicochemical properties have been designed. Owing to the unique characteristics, DNAs have been used as a functional building block for novel nanoarchitecture. In particular, a self-assembly of long DNA molecules via a piece DNA staple has been utilized to attain such constructs. However, it needs many talented prerequisites (e.g., complicated computer program) with fewer yields of products. In addition, it has many limitations to overcome: for instance, (i) thermal instability under moderate environments and (ii) restraint in size caused by the restricted length of scaffold strands. Alternatively, the enzymatic sewing linkage of short DNA blocks is simply designed into long DNA assemblies but it is more error-prone due to the undeveloped sequence data. Here, we present, for the first time, a comprehensive study for directly combining DNA structures into higher DNA sewing constructs through the 5'-end cohesive ligation of T4 enzyme. Inspired by these achievements, the synthesized DNA nanomaterials were also utilized for effective detection and real-time diagnosis of cancer-specific and cytosolic RNA markers. This generalized protocol for generic DNA sewing is expected to be useful in several DNA nanotechnology as well as any nucleic acid-related fields.

  20. Spectroscopic investigation of UV irradiated enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Ware, D.L.; Hibbard, L.B. (Spelman College, Atlanta, GA (United States))

    1993-01-01

    Trptophan (Trp) undergoes photolysis when exposed to light in the near UV region. The enzyme systems horse liver alcohol dehydrogenase (HLAD) and glyceraldehyde 3-phosphate dehydrogenase (G3PDH), which contain two and three Trps respectively, were chosen for analysis of Trp photolysis. Aqueous solutions of HLAD and G3PDH were irradiated at either 295 or 335nm with a xenon lamp. Tryptophan fluorescence was monitored at half hour intervals for two hours in the case of HLAD and one hour in the case of G3PDH. The decrease in fluorescence from 295 irradiation was compared to the fluorescence decrease from 335 irradiation and was found to be similar for both the HLAD and G3PDH samples irradiated at 295nm and for both samples irradiated at 335nm. It was found that, for HLAD, the 295 irradiation caused a decrease in fluorescence of 90% as compared to a decrease of only 12% of 335nm. Enzymatic assays were then performed to determine the enzymatic activity before and after irradiation.

  1. Production of Enzymes from Marine Actinobacteria.

    Science.gov (United States)

    Zhao, X Q; Xu, X N; Chen, L Y

    Marine actinobacteria are well recognized for their capabilities to produce valuable natural products, which have great potential for applications in medical, agricultural, and fine chemical industries. In addition to producing unique enzymes responsible for biosynthesis of natural products, many marine actinobacteria also produce hydrolytic enzymes which are able to degrade various biopolymers, such as cellulose, xylan, and chitin. These enzymes are important to produce biofuels and biochemicals of interest from renewable biomass. In this chapter, the recent reports of novel enzymes produced by marine actinobacteria are reviewed, and advanced technologies that can be applied to search for novel marine enzymes as well as for improved enzyme production by marine actinobacteria are summarized, which include ribosome engineering, genome mining, as well as synthetic biology studies. © 2016 Elsevier Inc. All rights reserved.

  2. Stabilized enzymes in continuous gas phase reactions

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Fangxiao; LeJeune, K.; Yang, Zhen [Univ. of Pittsburgh, PA (United States)] [and others

    1995-12-01

    We are assessing the utility of enzymes to catalyze reactions in a continuous gas phase reactor. First, alcohol dehydrogenase has been used to oxidize an unsaturated alcohol, 3-methyl-2-buten-1-ol (UOL), to the corresponding unsaturated aldehyde, 3-methyl-2-butenal (UAL). Cofactor NAD{sup +} was regenerated by concomitant acetone reduction to isopropyl alcohol. Second, organophosphorus hydrolase (OPH) has been used to hydrolyze pesticide vapors. In order to control enzyme hydration level, enzyme water adsorption isotherms at different temperature have been studied. Huttig`s isotherm model has been found suitable to describe adsorption behavior. The influence of enzyme hydration level, enzyme loading on glass beads, reaction temperature and flow rate on enzymatic reaction rate and biocatalyst stability were investigated. Reaction kinetics were studied and a kinetic model was proposed. We will also report our attempts to further stabilize enzymes for use in gas reactions by incorporating them into polymer matrices.

  3. Prediction of Wild-type Enzyme Characteristics

    DEFF Research Database (Denmark)

    Geertz-Hansen, Henrik Marcus

    of biotechnology, including enzyme discovery and characterization. This work presents two articles on sequence-based discovery and functional annotation of enzymes in environmental samples, and two articles on analysis and prediction of enzyme thermostability and cofactor requirements. The first article presents...... a sequence-based approach to discovery of proteolytic enzymes in metagenomes obtained from the Polar oceans. We show that microorganisms living in these extreme environments of constant low temperature harbour genes encoding novel proteolytic enzymes with potential industrial relevance. The second article...... presents a web server for the processing and annotation of functional metagenomics sequencing data, tailored to meet the requirements of non-bioinformaticians. The third article presents analyses of the molecular determinants of enzyme thermostability, and a feature-based prediction method of the melting...

  4. Effects of ammonia on processing and secretion of precursor and mature lysosomal enzyme from macrophages of normal and pale ear mice: evidence for two distinct pathways

    OpenAIRE

    1985-01-01

    Lysosomal enzymes have been shown to be synthesized as microsomal precursors, which are processed to mature enzymes located in lysosomes. We examined the effect of ammonium chloride on the intracellular processing and secretion of two lysosomal enzymes, beta-glucuronidase and beta-galactosidase, in mouse macrophages. This lysosomotropic drug caused extensive secretion of both precursor and mature enzyme forms within a few hours, as documented by pulse radiolabeling and molecular weight analys...

  5. Trace elements as an activator of antioxidant enzymes

    Directory of Open Access Journals (Sweden)

    Marta Wołonciej

    2016-12-01

    Full Text Available Oxidative stress is a state of impaired balance between the formation of free radicals and antioxidant capacity of the body. It causes many defects of the body, e.g. lipid peroxidation, DNA and protein damage. In order to prevent the effects of oxidative stress, the organism has developed defence mechanisms. These mechanisms capture and inhibit the formation of free radicals and also chelate ion metals that catalyse free radical reactions. Trace elements are components of antioxidant enzymes involved in antioxidant mechanisms. Selenium, as a selenocysteine, is a component of the active site of glutathione peroxidase (GPx. The main function of GPx is neutralization of hydrogen peroxide (H2O2 and organic peroxide (LOOH. Furthermore, selenium is a structural part of a large group of selenoproteins that are necessary for proper functioning of the body. Manganese, copper and zinc are a part of the group of superoxide dismutase enzymes (MnSOD, Cu/ZnSOD, which catalyse the superoxide anion dismutation into hydrogen peroxide and oxygen. Formed hydrogen peroxide is decomposed into water and oxygen by catalase or glutathione peroxidase. An integral component of catalase (CAT is iron ions. The concentration of these trace elements has a significant influence on the activity of antioxidant enzymes, and thus on defence against oxidative stress. Even a small change in the level of trace elements in the tissue causes a disturbance in their metabolism, leading to the occurrence of many diseases.

  6. Orthotopic Liver Transplantation for Urea Cycle Enzyme Deficiency

    Science.gov (United States)

    Todo, Satoru; Starzl, Thomas E.; Tzakis, Andreas; Benkov, Keith J.; Kalousek, Frantisek; Saheki, Takeyori; Tanikawa, Kyuichi; Fenton, Wayne A.

    2010-01-01

    Hyperammonemia, abnormalities in plasma amino acids and abnormalities of standard liver functions were corrected by orthotopic liver transplantation in a 14-day-old boy with carbamyl phosphate synthetase-I deficiency and in a 35-yr-old man with argininosuccinic acid synthetase deficiency. The first patient had high plasma glutamine levels and no measureable citrulline, whereas citrulline values were markedly increased in Patient 2. Enzyme analysis of the original livers showed undetectable activity of carbamyl phosphate synthetase-I in Patient 1 and arginosuccinic acid synthetase in Patient 2. Both patients were comatose before surgery. Intellectual recovery of patient 1 has been slightly retarded because of a brain abscess caused by Aspergillus infection after surgery. Both patients are well at 34 and 40 mo, respectively, after surgery. Our experience has shown that orthotopic liver transplantation corrects the life-threatening metabolic abnormalities caused by deficiencies in the urea cycle enzymes carbamyl phosphate synthetase-I and arginosuccinic acid synthetase. Seven other patients–six with ornithine transcarbamylase deficiency and another with carbamyl phosphate synthetase-I deficiency–are known to have been treated elsewhere with liver transplantation 1½ yr or longer ago. Four of these seven recipients also are well, with follow-ups of 1½ to 5 yr. Thus liver transplantation corrects the metabolic abnormalities of three of the six urea cycle enzyme deficiencies, and presumably would correct all. PMID:1544622

  7. The interaction of the Eco R1 restriction enzyme E.coli with nucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Hollis, Donald F. [Univ. of California, Berkeley, CA (United States)

    1979-11-01

    The Eco R1 restriction enzyme can be shown to be inhibited by nucleotides which correspond to any part of its known site of phosphodiesterase activity. A series of di-, tetra-, and hexa-nucleotide fragments were synthesized and their effect on the activity of the enzyme upon superhelical Co1 E1 DNA studied. The inhibition caused by the individual mononucleotides were also studied. In general all the nucleotide fragments showed some form of interaction with the enzyme system. Tetranucleotides were stronger inhibitors than dinucleotides, which in turn were stronger inhibitors than the mononucleotides. Within each category of inhibitors, those containing the phosphodiester bond which is acted upon by the enzyme were the strongest inhibitors. Only those fragments which were consistent with the enzymes site of activity showed competitive inhibition kinetics. Nucleotides which do not fit within the site of phosphodiesterase activity show non-competitive inhibition kinetics.

  8. A study of overproduction and enhanced secretion of enzymes. Quarterly report

    Energy Technology Data Exchange (ETDEWEB)

    Dashek, W.V.

    1993-09-01

    Wood decay within forests, a significant renewable photosynthetic energy resource, is caused primarily by Basidiomycetous fungi, e.g., white rot fungi. These organisms possess the ability to degrade lignin, cellulose and hemicellulose, the main organic polymers of wood. In the case of the white rot fungi, e.g., Coriolus versicolor, the capacity results from the fungus` ability to elaborate extracellular cellulolytic and ligninolytic enzymes. With regard to the latter, at least one of the enzymes, polyphenol oxidase (PPO) appears within a defined growth medium. This proposal focuses on the over-production and enhanced secretion of PPO, cellulase and lignin peroxidase. There are two major sections to the proposal: (1) overproduction of lignocellulolytic enzymes by genetic engineering methodologies and hyper-production and enhanced secretion of these enzymes by biochemical/electro microscopical techniques and (2) the biochemical/electron microscopical method involves substrate induction and the time-dependent addition of respiration and PPO enzymes.

  9. Reconstitution studies on the involvement of radiation-induced lipid peroxidation in damage to membrane enzymes.

    Science.gov (United States)

    Yukawa, O; Nagatsuka, S; Nakazawa, T

    1983-04-01

    The effect of radiation on the drug-metabolizing enzyme system of microsomes, reconstituted with liposomes of microsomal phospholipids, NADPH-cytochrome P-450 reductase and cytochrome P-450, was examined to elucidate the role of lipid peroxidation of membranes in radiation-induced damage to membrane-bound enzymes. The reconstituted system of non-irradiated enzymes with irradiated liposomes showed a low activity of hexobarbital hydroxylation, whereas irradiated enzymes combined with non-irradiated liposomes exhibited an activity equal to that of unirradiated controls. Irradiation of liposomes caused a decrease in cytochrome P-450 content by destruction of the haem of cytochrome P-450 and also inhibited the binding capacity of cytochrome P-450 for hexobarbital. The relationship between radiation-induced lipid peroxidation and membrane-bound enzymes is discussed.

  10. Reconstitution studies on the involvement of radiation-induced lipid peroxidation in damage to membrane enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Yukawa, O.; Nagatsuka, S.; Nakazawa, T. (National Inst. of Radiological Sciences, Chiba (Japan))

    1983-04-01

    The effect of ..gamma..-radiation on the drug-metabolizing enzyme system of microsomes, reconstituted with liposomes of microsomal phospholipids, NADPH-cytochrome P-450 reductase and cytochrome P-450, was examined to elucidate the role of lipid peroxidation of membranes in radiation-induced damage to membrane-bound enzymes. The reconstituted system of non-irradiated enzymes with irradiated liposomes showed a low activity of hexobarbital hydroxylation, whereas irradiated enzymes combined with non-irradiated liposomes exhibited an activity equal to that of unirradiated controls. Irradiation of liposomes caused a decrease in cytochrome P-450 content by destruction of the haem of cytochrome P-450 and also inhibited the binding capacity of cytochrome P-450 for hexobarbital. The relationship between radiation-induced lipid peroxidation and membrane-bound enzymes is discussed.

  11. Conductometric transducers for enzyme-based biosensors.

    Science.gov (United States)

    Mikkelsen, S R; Rechnitz, G A

    1989-08-01

    The use of alternating current conductometric transducers in biosensing devices has been investigated for urea and D-amino acid sensors using the enzyme systems urease and D-amino acid oxidase/catalase. Transducers with copper and platinum electrodes were constructed and characterized, and two enzyme immobilization methods were tested. Detection limits of 1 x 10(-6)M and linear ranges of 2 orders of magnitude were routinely achieved for these model sensors with enzymes covalently immobilized on collagen films.

  12. ENZYME RESISTANCE OF GENETICALLY MODIFIED STARCH POTATOES

    Directory of Open Access Journals (Sweden)

    A. Sh. Mannapova

    2015-01-01

    Full Text Available Here in this article the justification of expediency of enzyme resistant starch use in therapeutic food products is presented . Enzyme resistant starch is capable to resist to enzymatic hydrolysis in a small intestine of a person, has a low glycemic index, leads to decrease of postprandial concentration of glucose, cholesterol, triglycerides in blood and insulin reaction, to improvement of sensitivity of all organism to insulin, to increase in sense of fulness and to reduction of adjournment of fats. Resistant starch makes bifidogenшс impact on microflora of a intestine of the person, leads to increase of a quantity of lactobacillus and bifidobacterium and to increased production of butyric acid in a large intestine. In this regard the enzyme resistant starch is an important component in food for prevention and curing of human diseases such as diabetes, obesity, colitis, a cancer of large and direct intestine. One method is specified by authors for imitation of starch digestion in a human body. This method is based on the definition of an enzyme resistance of starch in vitro by its hydrolysis to glucose with application of a glucoamylase and digestive enzyme preparation Pancreatin. This method is used in researches of an enzyme resistance of starch, of genetically modified potato, high amylose corn starch Hi-Maize 1043 and HYLON VII (National Starch Food Innovation, USA, amylopectin and amylose. It is shown that the enzyme resistance of the starch emitted from genetically modified potatoes conforms to the enzyme resistance of the high amylose corn starch “Hi-Maize 1043 and HYLON VII starch”, (National Starch Food Innovation, the USA relating to the II type of enzyme resistant starch. It is established that amylopectin doesn't have the enzyme resistant properties. The results of researches are presented. They allow us to make the following conclusion: amylose in comparison with amylopectin possesses higher enzyme resistance and gives to

  13. 蕤核叶片解剖结构和组织化学定位研究%Anatomical Structure and Histochemical Localization of the Leaf of Prinsepia uniflora Batal

    Institute of Scientific and Technical Information of China (English)

    陈婧; 杨瑞林; 朱广龙; 张强; 魏学智

    2012-01-01

    采用解剖学和组织化学方法,研究了蕤核(Prinsepia uniflora Batal.)叶片的解剖结构,并对叶片中的黄酮类、生物碱类及多糖的组织化学定位进行了研究.结果表明:蕤核叶片上表皮有角质层,下表皮有气孔分布,气孔密度为278个·mm-2,近轴面栅栏组织细胞2~3层,排列紧密而整齐,含有许多晶体;叶片主脉木质部发达,由多列导管组成.上述特征说明蕤核叶片的解剖结构与环境之间的适应性.组织化学定位显示黄酮多分布于栅栏组织和厚角组织,生物碱含量少,多糖均匀分布于叶肉中.%The methods of anatomy and histochemistry were used to research the anatomical structure and histochemical localization of flavonoids, alkaloids and polysaccharides of the leaf of Prinsepia uniflora Batal. The results showed that the structure of the leaf of P. uniflora Batal was as follows: the upper epidermis cell with relative thick horny layer, the stomata existed in lower epidermis,the density of the stomata was 278 mm"2. The upper side included 2 to 3 layers of palisade tissue cells, with a lot of crystals; the main vein of leaves had well developed xylem, with a number of radial arranged vessels. The characteristics mentioned above showed that P. uniflora Batal adapted well to the environment. The histochemical localization showed that; the total flavonoids mainly accumulated in the palisade tissue and collenchyma, alkaloids were poor existed in the leaf, polysaccharides were generally distributed in mesophyll of P. uniflora Batal.

  14. Análise histoquímica foliar do amendoim: genótipos 'Tatu' e SO-909 Leaf histochemical analyses of peanut: genotypes 'Tatu' and SO-909

    Directory of Open Access Journals (Sweden)

    Renato Ferraz de Arruda Veiga

    1992-01-01

    Full Text Available Este trabalho teve por finalidade a análise histoquímica foliar de dois genótipos de amendoim (Arachis hypogaea L., do tipo botânico Valência: SO-53 ('Tatu' e SO-909 (PI-259747, cuja literatura demonstra apresentarem respostas diferentes de resistência às principais moléstias fúngicas foliares do Brasil. Seções transversais das seguintes estruturas - pulvino, haste peciolar, raque, pulvínulo e folíolo - e seções paradérmicas de folíolos coletados em dois anos agrícolas consecutivos, foram analisadas quanto à presença de alcalóides, amido, calose, celulose pura, celulose com pectina, cera, cristais, cutina, lignina, mucilagem, óleo, resina, tanino e ureídeos (micrograma por folíolo (grama. As diferenças qualitativas histoquímicas observadas nos diversos tecidos, como a freqüência de tanino, alcalóide, pectina e óleo, supostamente, podem ser responsáveis pela resistência ou suscetibilidade dos genótipos às moléstias fúngicas foliares. Para fins de caracterização, mostrou-se eficiente a avaliação de pureza de celulose.Leaf histochemical analyses were made in two genotypes of Arachis hypogaea L., of the Valencia group, which present different responses to some of the peanut foliar diseases. The analyses were performed on cross sections of the pulvini, petiole, rachis, pulvinulus and leaflets. The following constituents were observed: alkaloids, callose, cellulose with pectin, cristal, cutin, lignin, mucilage, oil, pure cellulose, resin, starch, tannin, wax and weight of ureides by leaflets. Some histochemical characteristics such the amount of tannin, alkaloids, pectin and oil can be produce different responses of peanut to foliar fungal diseases, and can be used in the characterization of peanut genotypes like the amount of pure cellulose and cellulose with pectin.

  15. Peptide-modified surfaces for enzyme immobilization.

    Directory of Open Access Journals (Sweden)

    Jinglin Fu

    Full Text Available BACKGROUND: Chemistry and particularly enzymology at surfaces is a topic of rapidly growing interest, both in terms of its role in biological systems and its application in biocatalysis. Existing protein immobilization approaches, including noncovalent or covalent attachments to solid supports, have difficulties in controlling protein orientation, reducing nonspecific absorption and preventing protein denaturation. New strategies for enzyme immobilization are needed that allow the precise control over orientation and position and thereby provide optimized activity. METHODOLOGY/PRINCIPAL FINDINGS: A method is presented for utilizing peptide ligands to immobilize enzymes on surfaces with improved enzyme activity and stability. The appropriate peptide ligands have been rapidly selected from high-density arrays and when desirable, the peptide sequences were further optimized by single-point variant screening to enhance both the affinity and activity of the bound enzyme. For proof of concept, the peptides that bound to β-galactosidase and optimized its activity were covalently attached to surfaces for the purpose of capturing target enzymes. Compared to conventional methods, enzymes immobilized on peptide-modified surfaces exhibited higher specific activity and stability, as well as controlled protein orientation. CONCLUSIONS/SIGNIFICANCE: A simple method for immobilizing enzymes through specific interactions with peptides anchored on surfaces has been developed. This approach will be applicable to the immobilization of a wide variety of enzymes on surfaces with optimized orientation, location and performance, and provides a potential mechanism for the patterned self-assembly of multiple enzymes on surfaces.

  16. The mechanisms of Excited states in enzymes

    DEFF Research Database (Denmark)

    Petersen, Frederic Nicolas Rønne; Bohr, Henrik

    2010-01-01

    Enzyme catalysis is studied on the basis of excited state processes, which are of electronic, vibrational and thermal nature. The ways of achieving the excited state, such as photo-absorption and ligand binding, are discussed and exemplified by various cases of enzymes.......Enzyme catalysis is studied on the basis of excited state processes, which are of electronic, vibrational and thermal nature. The ways of achieving the excited state, such as photo-absorption and ligand binding, are discussed and exemplified by various cases of enzymes....

  17. The mechanisms of Excited states in enzymes

    DEFF Research Database (Denmark)

    Petersen, Frederic Nicolas Rønne; Bohr, Henrik

    2010-01-01

    Enzyme catalysis is studied on the basis of excited state processes, which are of electronic, vibrational and thermal nature. The ways of achieving the excited state, such as photo-absorption and ligand binding, are discussed and exemplified by various cases of enzymes.......Enzyme catalysis is studied on the basis of excited state processes, which are of electronic, vibrational and thermal nature. The ways of achieving the excited state, such as photo-absorption and ligand binding, are discussed and exemplified by various cases of enzymes....

  18. Operating windows for enzyme enhanced PCCC

    DEFF Research Database (Denmark)

    Deslauriers, Maria Gundersen; Gladis, Arne; Fosbøl, Philip Loldrup

    2017-01-01

    Today, enzyme enhanced carbon capture and storage (CCS) is gaining interest, since it can enable the use of energy efficient solvents, and thus potentially reduce the carbon footprint of CCS. However, a limitation of this technology is the high temperatures encountered in the stripper column, which...... capture capacity of 1 MTonn CO2/year, and is here operated for one year continuously. This publication compares soluble enzymes dissolved in a capture solvent with and without the use of ultrafiltration membranes. The membranes used here have an enzyme retention of 90%, 99% and 99.9%. Enzyme retention...

  19. A quantitative histochemical study of lactate dehydrogenase and succinate dehydrogenase activities in the membrana granulosa of the ovulatory follicle of the rat.

    Science.gov (United States)

    Zoller, L C; Enelow, R

    1983-11-01

    Using a microdensitometer, lactate dehydrogenase and succinate dehydrogenase activities were measured in the membrana granulosa of the rat ovulatory follicle. Ovaries were removed on each day of the oestrous cycle; oestrus, dioestrus-1, dioestrus-2, and proestrus; and enzyme activities measured in the membrana granulosa as a whole and in four regions within it: peripheral (PR), antral (AR), cumulus oophorus (CO) and corona radiata (CR). Throughout the cycle, lactate dehydrogenase activity was greatest in PR. On oestrus, lactate dehydrogenase activity was progressively less in AR, CO and CR. On dioestrus-1, activity was identical in AR and CO and less in CR. On dioestrus-2, activity was greater in AR than in CO or CR. By proestrus, activity was equal in AR, CO and CR. In the membrana granulosa as a whole, and in each region, lactate dehydrogenase activity declined as ovulation approached. In contrast, succinate dehydrogenase activity in the membrana granulosa as a whole and in PR was constant throughout the cycle. Activity fluctuated in the other regions. Succinate dehydrogenase activity on oestrus was greatest in PR, less in AR and CO and least in CR. On the remaining days, succinate dehydrogenase activity was greatest in PR and less but equal in the remainder of the membrana granulosa.

  20. Experimental protein malnutrition as a causative factor in the histological and histochemical disruption of the ependymal cells of the third ventricla and cervical central canal of squirrel monkeys.

    Science.gov (United States)

    Sharma, S P; Manocha, S L

    1977-01-01

    The histologic disturbances of the cuboidal and columnar ependymal cells of the neonates under the extrinsic influence of maternal protein deprivation during most of the gestation period have been demonstrated in the central canal of the cervical spinal cord and III ventricle of the squirrel monkey brain. The control animals whose mothers were maintained on high protein diets showed an unbroken ependymal layer with an intact glial fiber layer and the subependymal cell plate. Taking birth-weight as an indication of the degree of malnutrition, maximum disruption of the ependymal layer was observed in those animals born around 80 g birth-weight. In the latter, most of the ependymal cells lost their characteristic arrangement and columnar shapes. The ependymal cells of the manourished animals show a marked reduction of oxidative enzyme content and relatively active glycogen metabolism as compared to the healthy controls. The significance of these changes has been discussed in the light of parallels in the embryological development and, to some extent, functions between the orinary neurons, ependymal cells of the choroid plexus, other ependymal cells and the glial cells. In this context, the changes observed in the present studies under the effect of protein malnutrition must be taken as part of overall changes experienced by various kinds of cells comprising the nervous system.