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Sample records for saginata metacestodes purified

  1. Usefulness of serological ELISA assay forTaenia saginata to detect naturally infected bovines

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    Silvana de Cássia Paulan

    Full Text Available Bovine cysticercosis, a cosmopolitan disease caused by Taenia saginata, leads to economic losses due to carcass devaluation at slaughter. Sanitary inspection at slaughterhouses, the routine diagnostic method in Brazil, lacks the necessary sensitivity to detect the mildly infected cattle that are typically encoutered in Brazil. In this study we have tested cattle sera from animals diagnosed as positive and negative by veterianry inspection for (1 anti-parasite antibodies using metacestodes antigens (T. solium vesicular fluid and T. saginata secretions and (2 the HP10 secreted antigen of viable metacestodes. The cut-off values were calculated by ROC curve for intense and mild infections conditions, and by the classical method ( for negative samples. The sensitivity and specificity of these diagnostic tests were different depending on the assumed cut-off value and, importantly, whether the infection was mild or intense. In spite of these observations, however, such ELISA assays for serum antibodies and parasite antigens constitute an important tool for epidemiological porposes, and in establishing priorities for the control of bovine cysticercosis.

  2. Experimental Infection of Taenia saginata eggs in Bali Cattle: Distribution and Density of Cysticercus bovis

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    Nyoman Sadra Dharmawan

    2009-12-01

    Full Text Available The objective of this study was to observe the development, distribution, and infection density ofTaenia saginata metacestodes in Bali cattle. Three Bali cattle were experimentally infected with T. saginataeggs which were collected from taeniasis patients. The experimental animal was inoculated with : i1000,00 T. saginata; ii 500,000 eggs; and iii 1,000,000 eggs, respectivelly 100,000 (cattle 1, 500,000(cattle 2, and 1,000,000 (cattle 3 T. saginata eggs, respectively. To observe the development of cysticerci,all cattle were slaughtered at 24 weeks post infection. To observe their distribution and density, slicingwas done to the cattle?s tissues. The study results showed that cysts were found distributed to all muscletissues and some visceral organs such as heart, diaphragm, lungs, and kidney of the cattle infected with100,000 and 500,000 T. saginata eggs. Density of the cyst was in the range of 11 to 95 cysts per 100 gramsof tissue. The highest density was noted in the heart (58/100 grams and in diaphragm (55/100 grams.This study has confirmed that T. saginata eggs derived from taeniasis patient in Bali, if infected to Balicattle can develop and spread to all muscle tissues and some visceral organs. From this study it wasconcluded that it is necessary to include the heart in the meat inspection at slaughter house for possibilityof T. saginata cyst infection.$?

  3. Usefulness of serological ELISA assay for Taenia saginata to detect naturally infected bovines Utilização de teste sorológico ELISA para a detecção de bovinos naturalmente infectados por Taenia saginata

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    Silvana de Cássia Paulan

    Full Text Available Bovine cysticercosis, a cosmopolitan disease caused by Taenia saginata, leads to economic losses due to carcass devaluation at slaughter. Sanitary inspection at slaughterhouses, the routine diagnostic method in Brazil, lacks the necessary sensitivity to detect the mildly infected cattle that are typically encoutered in Brazil. In this study we have tested cattle sera from animals diagnosed as positive and negative by veterianry inspection for (1 anti-parasite antibodies using metacestodes antigens (T. solium vesicular fluid and T. saginata secretions and (2 the HP10 secreted antigen of viable metacestodes. The cut-off values were calculated by ROC curve for intense and mild infections conditions, and by the classical method ( for negative samples. The sensitivity and specificity of these diagnostic tests were different depending on the assumed cut-off value and, importantly, whether the infection was mild or intense. In spite of these observations, however, such ELISA assays for serum antibodies and parasite antigens constitute an important tool for epidemiological porposes, and in establishing priorities for the control of bovine cysticercosis.A cisticercose bovina, uma doença cosmopolita causada pela Taenia saginata, resulta em perdas econômias devido à desvalorização de carcaças durante o abate. A inspeção sanitária nos frigoríficos, método de diagnóstico de rotina no Brasil, não possui sensibilidade necessária para detectar animais levemente infectados, os quais são tipicamente encontrados no Brasil. Neste estudo testou-se soro de animais diagnosticados positivos e negativos pela inspeção veterinária por (1 anticorpos anti-parasita usando antígenos de metacestóides (fluido vesicular de T. solium e secreções de T. saginata e (2 antígeno secretado de metacestóides viáveis. Os pontos de corte foram calculados pela curva ROC, considerando condições de intensa e leve infeção, e pelo método clássico ( das amostras

  4. Recent hybridization between Taenia asiatica and Taenia saginata.

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    Yamane, Kanako; Suzuki, Yumi; Tachi, Eiko; Li, Tiaoying; Chen, Xingwang; Nakao, Minoru; Nkouawa, Agathe; Yanagida, Testuya; Sako, Yasuhito; Ito, Akira; Sato, Hiroshi; Okamoto, Munehiro

    2012-06-01

    Five Taenia tapeworms collected from humans in Tibetan Plateau, Sichuan, China, where three species of human Taenia are sympatrically endemic, were examined for the mitochondrial cox1 gene and two nuclear genes, ef1 and elp. Phylogenetic analyses of these genes revealed that two adult worms showed nuclear-mitochondrial discordance, suggesting that they originated from hybridization between Taenia saginata and Taenia asiatica. One of two worms had T. asiatica-type mtDNA, whereas another worm had T. saginata-type mtDNA, indicating that reciprocal hybridization between T. saginata and T. asiatica could occur. The worm having T. asiatica-type mtDNA was heterozygous at both nuclear loci with T. saginata-type alleles and T. asiatica-type alleles. In another worm, the ef1 locus was heterozygous with a T. saginata-type alleles and T. asiatica-type alleles, while the elp locus was homozygous with T. saginata-type alleles. Self-fertilization is the main reproductive method of the genus Taenia. Since self-fertilization represents a type of inbreeding, each locus in the offspring would become homozygous over generations with genetic drift. The fact that some nuclear loci are still heterozygous means that hybridization might have occurred recently. Hybridization between T. asiatica and T. saginata is probably an ongoing event in many areas in which they are sympatrically endemic. Crown Copyright © 2012. Published by Elsevier Ireland Ltd. All rights reserved.

  5. TAENIA SAGINATA

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    Endang Setiyani

    2013-09-01

    Full Text Available Cacing pita ini adalah parasit dalam usus halusmanusia, dan sebagai inang antaranya adalah sapi.Penyakit infeksi yang disebabkan oleh cacing pitaTaenia saginata dikenal dengan nama Taeniasis. Infeksiterutama terjadi di Afrika, Timur Tengah, Eropa Barat,Meksiko, dan Amerika Selatan.

  6. Genotypic relationships between Taenia saginata, Taenia asiatica and their hybrids.

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    Yamane, Kanako; Yanagida, Tetsuya; Li, Tiaoying; Chen, Xingwang; Dekumyoy, Paron; Waikagul, Jitra; Nkouawa, Agathe; Nakao, Minoru; Sako, Yasuhito; Ito, Akira; Sato, Hiroshi; Okamoto, Munehiro

    2013-11-01

    Partial sequences of the DNA polymerase delta (pold) gene from Taenia saginata-like adult worms were sequenced. Phylogenetic analysis revealed that pold gene sequences were clearly divided into two clades, differing from each other in five to seven nucleotides. There is little doubt that T. saginata and Taenia asiatica were once separated into two distinct taxa as has been concluded in previous studies. On the other hand, most of the adult worms, which were identified as T. asiatica using mitochondrial DNA, were homozygous for an allele that originated from the allele of T. saginata via single nucleotide substitution. These results indicate that most of the adult worms, which had been called T. asiatica, are not actually 'pure T. asiatica' but instead originated from the hybridization of 'pure T. saginata' and 'pure T. asiatica'.

  7. Genetic diversity of Taenia saginata (Cestoda: Cyclophyllidea) from Lao People's Democratic Republic and northeastern Thailand based on mitochondrial DNA.

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    Sanpool, Oranuch; Rodpai, Rutchanee; Intapan, Pewpan M; Sadaow, Lakkhana; Thanchomnang, Tongjit; Laymanivong, Sakhone; Maleewong, Wanchai; Yamasaki, Hiroshi

    2017-03-11

    Taenia saginata is a tapeworm found in cattle worldwide. Analysis of genetic diversity in different geographical populations of T. saginata not only helps to understand the origin, transmission and spread of this organism, but also to evaluate the selection pressures acting on T. saginata and how it is responding to them. However, there are few reports of the genetic variability of T. saginata populations in different regions of the world, including Lao PDR and Thailand. We report the genetic diversity of T. saginata populations in Lao PDR and northeastern Thailand together with sequences of T. saginata from other countries deposited in GenBank. Mitochondrial cox1 sequence analysis revealed that 15 and 8 haplotypes were identified in 30 and 21 T. saginata isolates from Lao PDR and northeastern Thailand, respectively. Fifty-three haplotypes were identified from 98 sequences. Phylogenetic tree and haplotype network analyses revealed that global isolates of T. saginata were genetically divided into five groups (A, B, C1, C2 and D). Taenia saginata isolates from Lao PDR and northeastern Thailand belonged to either Group A or B. Taenia saginata from western Thailand clustered in groups C1, C2 and D, and populations from the northeast and western Thailand were found to be genetically distinct. Taenia saginata isolates in Lao PDR and Thailand were also found to be genetically diverse but the degree of genetic differentiation was low. Taenia saginata populations from Lao PDR and northeastern Thailand are genetically distinct from the population in western Thailand and it is proposed that T. saginata has been dispersed by different transmission routes in Southeast Asia.

  8. Echinococcus metacestode: in search of viability markers.

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    Gottstein, Bruno; Wang, Junhua; Blagosklonov, Oleg; Grenouillet, Frédéric; Millon, Laurence; Vuitton, Dominique A; Müller, Norbert

    2014-01-01

    Epidemiological studies have demonstrated that most humans infected with Echinococcus spp. exhibit resistance to disease. When infection leads to disease, the parasite is partially controlled by host immunity: in case of immunocompetence, the normal alveolar echinococcosis (AE) or cystic echinococcosis (CE) situation, the metacestode grows slowly, and first clinical signs appear years after infection; in case of impaired immunity (AIDS; other immunodeficiencies), uncontrolled proliferation of the metacestode leads to rapidly progressing disease. Assessing Echinococcus multilocularis viability in vivo following therapeutic interventions in AE patients may be of tremendous benefit when compared with the invasive procedures used to perform biopsies. Current options are F18-fluorodeoxyglucose-positron emission tomography (FDG-PET), which visualizes periparasitic inflammation due to the metabolic activity of the metacestode, and measurement of antibodies against recEm18, a viability-associated protein, that rapidly regresses upon metacestode inactivation. For Echinococcus granulosus, similar prognosis-associated follow-up parameters are still lacking but a few candidates may be listed. Other possible markers include functional and diffusion-weighted Magnetic Resonance Imaging (MRI), and measurement of products from the parasite (circulating antigens or DNA), and from the host (inflammation markers, cytokines, or chemokines). Even though some of them have been promising in pilot studies, none has been properly validated in an appropriate number of patients until now to be recommended for further use in clinical settings. There is therefore still a need to develop reliable tools for improved viability assessment to provide the sufficient information needed to reliably withdraw anti-parasite benzimidazole chemotherapy, and a basis for the development of new alternative therapeutic tools. © B. Gottstein et al., published by EDP Sciences, 2014.

  9. Experimental single and trickle infections of cattle with Taenia saginata: Studies of immunodiagnosis

    DEFF Research Database (Denmark)

    Bøgh, H.O.; Grønvold, J.; Maeda, G.E.

    1996-01-01

    Two groups of 12 calves were given either a single infection dose of 30,000 Taenia saginata eggs (group A) or trickle infected with 12 x 2500 T saginata eggs (group B). The calves were killed 16, 22 or 54 weeks after infection and the numbers of viable and dead parasites were counted by slicing...

  10. Efficacy of albendazole in combination with thymol against Echinococcus multilocularis protoscoleces and metacestodes.

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    María, Albani Clara; Celina, Elissondo María

    2014-12-01

    The larval (metacestode) stage of the tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE), a mainly hepatic disease characterized by continuous asexual proliferation of metacestodes by exogenous budding, resulting in the tumor-like, infiltrative growth of the parasite lesion. Current chemotherapeutical treatment of AE relies on the use of benzimidazoles, albendazole (ABZ) and mebendazole, but these drugs act parasitostatic rather than parasitocidal, and due to their low success rate they imply a lifelong application causing severe side effects. Thymol is one of the major components of the essential oils of Thymus and is a widely known anti-microbial agent. The aim of the present work was to compare the efficacy of albendazole (ABZ) and thymol separately or combined on E. multilocularis protoscoleces and metacestodes. For this purpose, microscopical examinations at different time points were carried out. Moreover the tegumentary enzyme gamma glutamyl transferase (GGT) was measured to quantify the damage in metacestodes. Even though treatments of in vitro cultured E. multilocularis protoscoleces or metacestodes with ABZ or/and thymol showed that the drugs have an adverse effect on parasite viability, the combination of the two compounds at the concentration of 10μg/ml showed the maximum anti-parasitic effect. Three days postincubation the first effects of the treatment were detected on protoscoleces and a marked reduction in viability (33%) was registered at day 18. Incubation of E. multilocularis metacestodes in the presence of ABZ 10μg/ml+thymol 10μg/ml during 10 days resulted in dramatic alterations such as strongly irregular and fissured surface and markedly disrupted vesicles. Scanning electron microscopy showed that protoscoleces as well as the germinal layer of E. multilocularis metacestodes were dramatically damaged following ABZ or/and thymol treatment. Also an important increase of tegumentary enzyme GGT was registered after 72h

  11. Genetic polymorphism in Taenia solium metacestodes from different Brazilian geographic areas

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    Ivanildes Solange da Costa Barcelos

    2012-02-01

    Full Text Available The aim of the present study is to investigate genetic polymorphisms in Taenia solium metacestodes from different Brazilian geographical areas and to relate them to antibody recognition in serum samples of neurocysticercosis (NC patients. Metacestodes were obtained from the Distrito Federal (DF, Bahia, Minas Gerais (MG and São Paulo (SP regions of Brazil. Samples of human sera from 49 individuals with NC, 68 individuals with other helminthiasis and 40 healthy volunteers were analysed (157 individuals in total. Antigens were prepared and used in enzyme-linked immunosorbent assay and western blotting assays to detect specific immunoglobulin G antibodies. Genetic distances between metacestode populations were analysed using random amplified polymorphic DNA (RAPD analysis. Our results show that there was a higher frequency of reactivity in the DF region in the sera from NC patients (p < 0.05, while discrimination between active and inactive NC was seen only in extracts from the MG and SP regions (p < 0.05. Using RAPD, the sample from the DF region presented a greater increase compared to the other regions. A relationship between genetic polymorphisms among T. solium metacestodes from different areas in Brazil and the differences in antibody detection in patients with NC were established.

  12. Protein Spesifik Cairan Kista Cysticercus bovis pada Sapi Bali yang Diinfeksi dengan Taenia saginata (SPECIFIC PROTEIN OF CYSTICERCUS BOVIS CYST FLUID ON BALI CATTLE EXPERIMENTALLY INFECTED WITH TAENIA SAGINATA

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    Nyoman Sadra Dharmawan

    2013-08-01

    Full Text Available Cysticercus bovis is the larval stage of Taenia saginata, the bovine tapeworm. The infection of thislarval in cattle musculature causes Bovine cysticercosis or Cysticercosis bovis.  Bovine cysticercosis is foundworldwide, but mostly in developing countries, where unhygienic conditions, poor cattle managementpractices, and the absence of meat inspection are common.  The adult Taenia infection in man is referredto as taeniasis.  Taenia saginata taeniasis is also found almost all over the world.  The prevalence ofTaenia saginata taeniasis has reported up to 27.5% in Gianyar Bali. In order to control the diseases,vaccination against the larvae stages in cattle of Taenia saginata may play an important role in controllingthe disease in the endemic regions.  The aims of the present study were to prepare and to investigate theimmunogenic protein as vaccine candidate for controlling  Cysticercus bovis infection in in Bali cattle.Cysticercus protein from the cyst fluid was firstly used to immunize mice and the mice sera were thencollected. Cysticercus proteins then analyzed using sodium dodecyl sulfate-gel electrophoresis (SDS-PAGE.All cysticercus proteins were then visualized by Commasie blue staining. The proteins were also transferredonto nitrocellulose membrane and the immunogenic proteins were visualized by Western Blotting usingimmune sera raised in mice.  By Commasie blue staining, a total of 17 proteins were detected with themolecular weight of 14,86 kDa -122,40 kDa from the smallest to the largest. As many as 7 immunogenicproteins with the molecular weights of 16.81 kDa; 19.22 kDa; 20.98 kDa; 27.41 kDa; 34.02 kDa; 38.31 kDa;and 54.94kDa were detected.

  13. Control of Taenia saginata by post-mortem examination of carcasses

    African Journals Online (AJOL)

    Background: A study to curb transmission cycle of a zoonotic Taema cestodiasis between humans and cattle is presented. Objective: To evaluate the reliability of meat inspection procedure in detecting carcasses of cattle with T. saginata cysticercosis. Methods: A total of 55 cattle divided into two groups of artificially (n =30) ...

  14. Effects of X-ray on the metacestodes of Echinococcus granulosus in vitro.

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    Mao, Rui; Wu, Ge; Wang, Hui; Lu, Pengfei; Li, Jun; Li, Haitao; Ainiwaer, Aimudula; Bai, Yiwei; Shu, Mingyang; Bao, Yongxing; Zhang, Wenbao

    2017-09-21

    Radiotherapy may represent an alternative treatment modality for cystic echinococcosis (CE), but there is no adequate evidence for it up to now. In this study, we aim to investigate the parasiticidal effects of X-ray on the metacestodes of Echinococcus granulosus in vitro. Protoscoleces obtained from sheep naturally infected with CE were cultivated in RPMI 1640 medium containing 10% fetal bovine serum (FBS) at 37 °C in 5% CO 2 . Upon encystation on day 14, the metacestodes were subjected to various intensities of X-ray. Metacestode structures were observed using light microscope and transmission electron microscopy (TEM), and Real-Time PCR was carried out to determine the expression of EgTPX, EgHSP70, EgEPC1 and Caspase-3. On day 14, encystation was noticed in the majority of protoscoleces in the control group. In the X-ray groups, the encystation rate showed significant decrease compared with that of the control group (P granulosus. Irradiation triggered increased expression of EgTPX, EgHSP70, EgEPC1 and Caspase-3.

  15. Similar diagnostic performance for neurocysticercosis of three glycoprotein preparations from Taenia solium metacestodes.

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    Villota, Guido E; Gomez, Diana I; Volcy, Michel; Franco, Andrés F; Cardona, Edgar A; Isaza, Rodrigo; Sanzón, Fernando; Teale, Judy M; Restrepo, Blanca I

    2003-03-01

    The detection of antibodies to Taenia solium metacestodes is very important in the differential diagnosis of neurocysticercosis (NCC). In this study, an electroimmunotransfer blot (EITB) assay that uses an elaborate protocol with metacestode glycoproteins as antigens was compared with two other Western blots that use glycoproteins obtained using simpler methods, including an eluate from a lectin column, or the vesicular fluid (VF) of the parasite. The concordance between the three assays was 91% in patients with active NCC and 100% in patients with suspected NCC and previous documentation of negative serology. The specificities for the Western blots and the EITB assay were 98% and 100%, respectively (98% concordance). These data suggest that the simplest of these immunoassays, the one that uses the VF of T. solium metacestodes in a Western blot format, can be reliably used for the serologic diagnosis of NCC in developing countries where access to the EITB assay is difficult.

  16. Performance of the ELISA test for the diagnosis of cysticercosis using experimentally and naturally cattle infected with metacestode of Taenia saginata

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    Rafaella Paola Meneguete dos Guimarães-Peixoto

    2015-04-01

    Full Text Available The bovine taeniasis-cysticercosis complex can be defined as a set of pathological changes caused by the adult form of Taenia saginata in humans and their larval form in animals (Cysticercus bovis. Data on the occurrence of bovine cysticercosis comes from the records of veterinary inspection of meat in slaughterhouses under fiscalization, where some positive cases may go unnoticed, especially in moderate infections. So, it is relevant the use of serological tests that have greater sensitivity than the post-mortem routine exams. Studies have shown the possible application of the ELISA test as a tool for epidemiological studies of the parasitosis and in the identification of animals with cysts. The aim of this study is to determine the detection threshold of the indirect ELISA test, using experimentally and naturally infected animals in detecting cases of cysticercosis. The sensitivity of the test to naturally infected animals using a cut-off 1 and 2 added with 2 standart-deviation (SD was of 12% and 24.4%, respectively. However, when using the cut-off of 1 and 2 added with 3 SD, the test sensitivity dropped, represented by 14.4% and 1.99% of the test sensitivity. As for the samples from experimentally infected animals, using the cut-off 1 and 2 added with 2 SD the sensitivity was 55.9% and 92.5%; and adding 3 SD the values found were 31.2% and 86%, respectively. The specificity of the test in all situations tested was 100%. It is important to take into account the right choice of control sera used in the ELISA test, whereas, according to its application, it is necessary to increase the sensitivity or specificity.

  17. Proteomic analysis of Taenia solium metacestode excretion-secretion proteins.

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    Victor, B.; Kanobana, K.; Gabriel, S.; Polman, K.; Deckers, N.; Dorny, P.; Deelder, A.M.; Palmblad, M.

    2012-01-01

    The metacestode larval stage of Taenia solium is the causal agent of a zoonotic disease called cysticercosis. The disease has an important impact on pork trade (due to porcine cysticercosis) and public health (due to human neurocysticercosis). In order to improve the current diagnostic tools and to

  18. Taenia solium metacestode preparation in rural areas of sub ...

    African Journals Online (AJOL)

    Background: Taenia solium metacestodes/cysts obtained from pig carcasses constitute a primary source for diagnostic tools used for the detection of human cysticercosis. Data on T. solium cyst preparation in Africa is still scarce but required to establish independent reference laboratories. Objectives: The aim of the present ...

  19. Pig as a Favorable Animal for Taenia Saginata Asiatica Infection

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    Ping-Chin Fan

    2006-01-01

    Full Text Available The epidemiology of Taenia saginata in some parts of Asia is confusing, in that beef does not appear to be the source of infection. In some areas, beef is either not available or not eaten raw, whereas pork at times is eaten uncooked. In light of this situation, we have exposed pigs and other animals to infection with strains of T. saginata to establish their ability to serve as intermediate hosts. Eggs of Taiwan Taenia, Korea Taenia, Indonesia Taenia, Thailand Taenia, Philippines Taenia, Ethiopia Taenia, and Madagascar Taenia were fed to 83 pigs of three strains: 43 Small-Ear Miniature (SEM, 34 Landrace Small-Ear Miniature (L-SEM, and 6 Duroc-Yorkshire-Landrace (DYL. We also fed the eggs to 10 Holstein calves, 17 Sannean goats, and 4 monkeys (Macaca cyclopis. We succeeded in infecting SEM (infection rate 88%, cysticercus recovery rate 19.1%, L-SEM (83%, 1.1%, and DYL (100%, 0.3% pigs with Taiwan Taenia; SEM (100%, 1.7%, L-SEM (100%, 5.6%, and DYL (100%, 0.06% pigs with Korea Taenia; SEM (100%, 22% and L-SEM (100%, 1.6% pigs with Indonesia Taenia; SEM (75%, 0.06% pigs with Thailand Taenia SEM (100%, 11% pigs with Philippines Taenia; SEM (80%, 0.005% pigs with Ethiopia Taenia; SEM (100%, 0.2% pigs with Madagascar Taenia. Holstein calves became infected with Taenia from Taiwan (100%, 1.1%, Korea (100%, 0.03%, Thailand (100%, 0.2%, and the Philippines (100%, 6%; however, the cysticerci of Taenia from Korea, Thailand, and the Philippines were degenerated and/or calcified. Sannean goats became infected with Taenia from Taiwan (33%, 0.01% and Korea (50%, 0.02%, while monkeys became infected with Taenia from Taiwan (50%, 0.01%. However, the cysticerci were degenerated and/or calcified. Therefore, these strains of pig seem to be favorable animal models for experimental studies of T. saginata-like tapeworms, with the SEM pig the most favorable.

  20. Failure to protect calves against Taenia saginata using antigens prepared from in vitro cultivation of the larval stage.

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    Mitchell, G B; Armour, J

    1980-11-01

    Calves were vaccinated intramuscularly against the tapeworm Taenia saginata using excretory/secretory (ES) antigens from short and long term periods of in vitro cultivation of the larval stage of the parasite, four weeks before challenge with 5000 T saginata onchospheres. Neither immunisation regime employed afforded significant protection against challenge. It was considered that this may have been due to a reduction in concentration of, or detrimental effects to, potential immunogens during vaccine production. Elucidation of the nature of the protective ES antigens necessary for standardization of the technique has yet to be achieved in helminths.

  1. Profound activity of the anti-cancer drug bortezomib against Echinococcus multilocularis metacestodes identifies the proteasome as a novel drug target for cestodes.

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    Britta Stadelmann

    2014-12-01

    Full Text Available A library of 426 FDA-approved drugs was screened for in vitro activity against E. multilocularis metacestodes employing the phosphoglucose isomerase (PGI assay. Initial screening at 20 µM revealed that 7 drugs induced considerable metacestode damage, and further dose-response studies revealed that bortezomib (BTZ, a proteasome inhibitor developed for the chemotherapy of myeloma, displayed high anti-metacestodal activity with an EC50 of 0.6 µM. BTZ treatment of E. multilocularis metacestodes led to an accumulation of ubiquinated proteins and unequivocally parasite death. In-gel zymography assays using E. multilocularis extracts demonstrated BTZ-mediated inhibition of protease activity in a band of approximately 23 kDa, the same size at which the proteasome subunit beta 5 of E. multilocularis could be detected by Western blot. Balb/c mice experimentally infected with E. multilocularis metacestodes were used to assess BTZ treatment, starting at 6 weeks post-infection by intraperitoneal injection of BTZ. This treatment led to reduced parasite weight, but to a degree that was not statistically significant, and it induced adverse effects such as diarrhea and neurological symptoms. In conclusion, the proteasome was identified as a drug target in E. multilocularis metacestodes that can be efficiently inhibited by BTZ in vitro. However, translation of these findings into in vivo efficacy requires further adjustments of treatment regimens using BTZ, or possibly other proteasome inhibitors.

  2. Proteomic analysis of Taenia ovis metacestodes by high performance liquid chromatography-coupled tandem mass spectrometry.

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    Zheng, Yadong

    2017-03-15

    Taenia ovis metacestodes reside in the muscle of sheep and goats, and may cause great economic loss due to condemnation of carcasses if not effectively controlled. Although advances have been made in the control of T. ovis infection, our knowledge of T. ovis biology is limited. Herein the protein profiling of T. ovis metacestodes was determined by liquid chromatography-linked tandem mass spectrometry. A total of 966 proteins were identified and 25.1% (188/748) were annotated to be associated with metabolic pathways. Consistently, GO analysis returned a metabolic process (16.27%) as one of two main biological process terms. Moreover, it was found that 24 proteins, including very low-density lipoprotein receptor, enolase, paramyosin and endophilin B1, were abundant in T. ovis metacestodes. These proteins may be associated with motility, metabolism, signaling, stress, drug resistance and immune responses. Furthermore, comparative analysis of 5 cestodes revealed the presence of Taenia-specific enolases. These data provide clues for better understanding of T. ovis biology, which is informative for effective control of infection. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Apoptosis of mouse hippocampal cells induced by Taenia crassiceps metacestode factor.

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    Zepeda, N; Solano, S; Copitin, N; Chávez, J L; Fernández, A M; García, F; Tato, P; Molinari, J L

    2017-03-01

    Seizures, headache, depression and neurological deficits are the signs and symptoms most frequently reported in human neurocysticercosis. However, the cause of the associated learning and memory deficits is unknown. Here, we used Taenia crassiceps infection in mice as a model of human cysticercosis. The effects of T. crassiceps metacestode infection or T. crassiceps metacestode factor (MF) treatment on mouse hippocampal cells were studied; control mice were included. At 45 days after infection or treatment of the mice with MF, all mice were anaesthetized and perfused transcardially with saline followed by phosphate-buffered 10% formalin. Then the brains were carefully removed. Coronal sections stained using several techniques were analysed. Extensive and significant apoptosis was found in the experimental animals, mainly in the dentate gyrus, CA1, CA2, CA3 and neighbouring regions, in comparison with the apparently intact cells from control mice (P < 0.01). These results suggest that neurological deficits, especially the learning and memory deficits, may be generated by extensive apoptosis of hippocampal cells.

  4. Prevalensi infeksi taeniasis saginata pada konsumen lawar sapi di Kecamatan Sukawati, Kabupaten Gianyar tahun 2012

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    Oka Harimbawa

    2014-02-01

    Full Text Available Research indicates that taeniasis and cysticocircosis is endemic in Bali with increasing trend found each year. Previous research in Gianyar Regency indicated that taeniasis prevalence was 23.8%, with the highest prevalence found in the sub-district of Sukawati. This high prevalence is partly influenced by the community practices and a culture of raw beef consumption in this area. This research used a combination of quantitative and qualitative methods. Quantitative data was collected through cross sectional analysis of 80 samples of raw beef consumers (interviews on risk behaviours and past experience. Fecal samples from these respondents were also taken and examined for taeniasis saginata infection. Qualitative data was collected through participatory observation and in-depth interviews with raw beef foodstall owners. The research findings indicated that the percentage of taeniasis saginata infection was found predominantly in the 15-44 age group, were male and did not have a formal education. Bivariate analysis indicated that the most significant factors were gender (p=0.018, location of foodstall (p=0.001 and respondent lawar consumption frequency (p=0.013. Multivariate analysis using cox regression methods found that the location of foodstall had a significant correlation with the occurence of taenaisis saginata (p=0.003. Recommendations include education for raw beef providers and upscaled testing of consumers and further research into other variables such as livestock slaughter behaviours and meat preparation protocols.

  5. Taeniasis/cysticercosis in Bali, Indonesia.

    Science.gov (United States)

    Wandra, Toni; Sudewi, A A Raka; Swastika, I Kadek; Sutisna, Putu; Dharmawan, Nyoman S; Yulfi, Hemma; Darlan, Dewi Masyithah; Kapti, I Nengah; Samaan, Gina; Sato, Marcello Otake; Okamoto, Munehiro; Sako, Yasuhito; Ito, Akira

    2011-07-01

    Taenia solium and Taenia saginata are found in humans in Bali, Indonesia. During a field survey of 660 people in Bali from 2002-2009 of taeniasis/cysticercosis cases using mitochondrial DNA confirmation of the species, we detected 80 cases of T. saginata taeniasis, 2 dual T. saginata/T. solium infections with T. solium metacestodes in the brain and 12 neurocysticercosis (NCC) cases at Sanglah Hospital, Denpasar. Although the prevalence of NCC in Bali is low, sporadic cases are still present. There is no Taenia asiatica in Bali. We summarize here the field survey findings of taeniasis, including 1 dual infection with taeniasis and cysticercosis in 2007, and the reason why there are no T. asiatica cases and we describe 3 NCC cases admitted to Sanglah Hospital, Denpasar, Bali in 2004. Diagnosis was based on anamnesis, clinical examination, including CT Scan, histopathological, serological and mitochondrial DNA examinations. In order to prevent unexpected symptomatic NCC after treatment with praziquantel, we recommend introducing a rapid test to confirm taeniasis carriers and cysticercosis cases as a tool for real time diagnosis.

  6. Taenia solium, Taenia saginata, Taenia asiatica, their hybrids and other helminthic infections occurring in a neglected tropical diseases' highly endemic area in Lao PDR

    Science.gov (United States)

    Sato, Megumi; Yanagida, Tetsuya; Waikagul, Jitra; Pongvongsa, Tiengkham; Sako, Yasuhito; Sanguankiat, Surapol; Yoonuan, Tipparayat; Kounnavang, Sengchanh; Kawai, Satoru; Ito, Akira; Okamoto, Munehiro; Moji, Kazuhiko

    2018-01-01

    Most part of Southeast Asia is considered endemic for human-infecting Taenia tapeworms; Taenia solium, T. saginata, and T. asiatica. However, until now there was no report of the occurrence of human cases of T. asiatica in Lao PDR. This study, conducted in Savannakhet Province, Lao PDR, microscopically examined a total of 470 fecal samples by Kato Katz method and found 86% of people harboring at least one helminth. Hookworms were detected in 56% of the samples besides Opisthorchis like eggs (42%), Trichuris trichiura (27%), Ascaris spp. (14%), and Taenia spp. (4%) eggs. Serology for cysticercosis showed 6.8% positives with results varying from 3% to 14.3% in Ethnic School students and Kalouk Kao village respectively. Species-specific PCR targeting mitochondrial DNA (mtDNA) of 28 tapeworms, recovered from 16 patients, revealed T. solium (n = 2), T. saginata (n = 21), and T. asiatica (n = 5). Two patients were confirmed to be coinfected with T. saginata and T. asiatica, indicating the endemicity of the 3 human Taenia in Lao PDR. However, nucleotide sequencing of a nuclear DNA gene, DNA polymerase delta (pold) revealed that all the tapeworms identified as T. asiatica using mtDNA had T. saginata type allele at pold locus, demonstrating that they are not “pure T. asiatica” but the hybrid descendants between the two species, confirming the wide distribution of hybrids of T. saginata/ T. asiatica in Southeast Asia. The high prevalence of several helminthic NTDs in east Savannakhet area even with conventional control measures indicates the importance to establish wide and multifaceted health programs to sustainably improve the quality of life of the populations living in these communities. PMID:29420601

  7. Taenia solium, Taenia saginata, Taenia asiatica, their hybrids and other helminthic infections occurring in a neglected tropical diseases' highly endemic area in Lao PDR.

    Directory of Open Access Journals (Sweden)

    Marcello Otake Sato

    2018-02-01

    Full Text Available Most part of Southeast Asia is considered endemic for human-infecting Taenia tapeworms; Taenia solium, T. saginata, and T. asiatica. However, until now there was no report of the occurrence of human cases of T. asiatica in Lao PDR. This study, conducted in Savannakhet Province, Lao PDR, microscopically examined a total of 470 fecal samples by Kato Katz method and found 86% of people harboring at least one helminth. Hookworms were detected in 56% of the samples besides Opisthorchis like eggs (42%, Trichuris trichiura (27%, Ascaris spp. (14%, and Taenia spp. (4% eggs. Serology for cysticercosis showed 6.8% positives with results varying from 3% to 14.3% in Ethnic School students and Kalouk Kao village respectively. Species-specific PCR targeting mitochondrial DNA (mtDNA of 28 tapeworms, recovered from 16 patients, revealed T. solium (n = 2, T. saginata (n = 21, and T. asiatica (n = 5. Two patients were confirmed to be coinfected with T. saginata and T. asiatica, indicating the endemicity of the 3 human Taenia in Lao PDR. However, nucleotide sequencing of a nuclear DNA gene, DNA polymerase delta (pold revealed that all the tapeworms identified as T. asiatica using mtDNA had T. saginata type allele at pold locus, demonstrating that they are not "pure T. asiatica" but the hybrid descendants between the two species, confirming the wide distribution of hybrids of T. saginata/ T. asiatica in Southeast Asia. The high prevalence of several helminthic NTDs in east Savannakhet area even with conventional control measures indicates the importance to establish wide and multifaceted health programs to sustainably improve the quality of life of the populations living in these communities.

  8. Characterisation of the carbohydrate components of Taenia solium metacestode glycoprotein antigens.

    Science.gov (United States)

    Restrepo, B I; Obregón-Henao, A; Mesa, M; Gil, D L; Ortiz, B L; Mejía, J S; Villota, G E; Sanzón, F; Teale, J M

    2000-05-01

    Human neurocysticercosis is caused by Taenia solium metacestodes. It usually affects the central nervous system of humans and can be confused with other brain pathologies. The Lens culinaris-binding glycoproteins from this parasite have been shown to be ideal targets for the development of a highly specific immunoassay for the diagnosis of neurocysticercosis. In the present study we characterised the carbohydrates associated with five antigenic glycoproteins of T. solium metacestodes in the range of 12-28 kilodaltons. Lectin-affinities and enzymatic deglycosylations suggested that each of the five antigens contain various glycoforms of asparagine-linked carbohydrates of the hybrid, complex and probably high mannose type. These carbohydrates accounted for at least 30-66% of the apparent molecular mass of the glycoconjugates. In contrast, there was no evidence for the presence of O-linked carbohydrates. Lectin affinity patterns suggested that the sugars are short and truncated in their biosynthetic route, and that some contain terminal galactose moieties. Elucidating the precise structure of the carbohydrates and establishing their role in antigenicity will be essential to design strategies to produce them in large and reproducible amounts for the development of improved immunoassays.

  9. Taenia saginata Goeze, 1872 (Platyhelminthes: Cestoda: Taeniidae en un indígena de Tierra del Fuego, Chile

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    Eduardo I. Faúndez

    2015-06-01

    Full Text Available Se entrega el primer registro del céstodo Taenia saginata en un indígena de Tierra del Fuego (Patagonia chilena, correspondiente al año 1902. Se discute la importancia del registro y se entregan ilustraciones del ejemplar.

  10. About human taeniasis and Taenia saginata diagnosis by endoscopy.

    Science.gov (United States)

    Galán Puchades, María Teresa

    2016-10-01

    La carta al editor se refiere al artículo de Canaval-Zuleta et al. aceptado para publicación, titulado "Endoscopy as an alternative diagnostic and therapeutic technique for Taenia saginata". El trabajo presenta una serie de incorrecciones que deben ser aclaradas, o al menos parte de ellas en solo 300 palabras. La información sobre las vias de infeccion en taeniasis, así como la patogenia y técnicas de diagnóstico, no se ajustan a la realidad. Asimismo, ya está publicado que el diagnóstico por endoscopia es una técnica muy sensible pero nada específica, pues no permite distinguir entre las 3 especies humanas del género Taenia.

  11. Morphometric characteristics of the metacestode Echinococcus vogeli Rausch & Bernstein, 1972 in human infections from the northern region of Brazil.

    Science.gov (United States)

    Almeida, F; Oliveira, F; Neves, R; Siqueira, N; Rodrigues-Silva, R; Daipert-Garcia, D; Machado-Silva, J R

    2015-07-01

    Polycystic echinococcosis, caused by the larval stage (metacestode) of the small-sized tapeworm, Echinococcus vogeli, is an emerging parasitic zoonosis of great public health concern in the humid tropical rainforests of South and Central America. Because morphological and morphometric characteristics of the metacestode are not well known, hydatid cysts from the liver and the mesentery were examined from patients following surgical procedures. Whole mounts of protoscoleces with rostellar hooks were examined under light and confocal laser scanning microscopy. Measurements were made of both large and small hooks, including the total area, total length, total width, blade area, blade length, blade width, handle area, handle length and handle width. The results confirmed the 1:1 arrangement of hooks in the rostellar pad and indicated, for the first time, that the morphometry of large and small rostellar hooks varies depending upon the site of infection. Light and confocal microscopy images displayed clusters of calcareous corpuscles in the protoscoleces. In conclusion, morphological features of large and small rostellar hooks of E. vogeli are adapted to a varied environment within the vertebrate host and such morphological changes in calcareous corpuscles occur at different stages in the maturation of metacestodes.

  12. Taeniasis caused by Taenia saginata in Gianyar town and Taenia solium in Karangasem villages of Bali, Indonesia, 2011-2016: How to detect tapeworm carriers, anamnesis or microscopy?

    Science.gov (United States)

    Swastika, Kadek; Wandra, Toni; Dharmawan, Nyoman Sadra; Sudarmaja, I Made; Saragih, John Master; Diarthini, Luh Putu Eka; Ariwati, Luh; Damayanti, Putu Ayu Asri; Laksemi, Dewa Ayu Agus Sri; Kapti, Nengah; Sutisna, Putu; Yanagida, Tetsuya; Ito, Akira

    2017-10-01

    From January 2011 until September 2016, screening of taeniasis carriers was carried out in a town in Gianyar District (Taenia saginata) and in villages which consisted of several Banjars (the smallest community units) on the eastern slope of Mt. Agung, Karangasem District (Taenia solium) in Bali, Indonesia. Fecal samples from all community members who chose to participate were examined microscopically for detection of taeniid eggs each person completedwith a questionnaire to determine if they had seen whitish, noodle-like proglottids (anamnesis) in their feces. Members with egg positive feces, and those with anamnesis, were treated with niclosamide (Yomesan ® , Bayer). A total of 39T. saginata tapeworm carriers were confirmed in Gianyar after deworming based on anamnesis (100%, 39/39). Only three of them (3/39, 7.7%) and 3/173 participants (1.7%) were identified by fecal microscopy. In contrast, 20T. solium carriers including one migrated to Gianyar were confirmed from 12 patients with eggs in their feces and from another 8 persons of 12 persons suspected to be infected due anamnesis only (8/12,66.7%) in Karangasem. The majority of carriers (12/20, 60.0%) identified by microscopy included 4 (33.3%) and 8 (66.7%) carriers confirmed microscopically with and without anamnesis, respectively. The prevalence rate was 12/1090 (1.10%) of participants. The results indicate that anamnesis is reliable for detection of T. saginata carriers, whereas it is not so reliable for detection of T. solium taeniasis (8/12, 66.7%) and that microscopy is more informative than anamnesis for T. solium. Eggs were detected more frequently in T. solium carriers (4/12, 33.3%) than in patients infected with T. saginata (3/39, 7.7%). T. solium carriers have so far been confirmed from nine of 13 Banjars examined in Karangasem. This study reveals that anamnesis is highly useful for screening of T. saginata carriers, whereas microscopy is a more valuable tool for detection of T. solium carriers

  13. Activities of fenbendazole in comparison with albendazole against Echinococcus multilocularis metacestodes in vitro and in a murine infection model.

    Science.gov (United States)

    Küster, Tatiana; Stadelmann, Britta; Aeschbacher, Denise; Hemphill, Andrew

    2014-04-01

    The current chemotherapeutic treatment of alveolar echinococcosis (AE) in humans is based on albendazole and/or mebendazole. However, the costs of treatment, life-long consumption of drugs, parasitostatic rather than parasiticidal activity of chemotherapy, and high recurrence rates after treatment interruption warrant more efficient treatment options. Experimental treatment of mice infected with Echinococcus multilocularis metacestodes with fenbendazole revealed similar efficacy to albendazole. Inspection of parasite tissue from infected and benzimidazole-treated mice by transmission electron microscopy (TEM) demonstrated drug-induced alterations within the germinal layer of the parasites, and most notably an almost complete absence of microtriches. On the other hand, upon in vitro exposure of metacestodes to benzimidazoles, no phosphoglucose isomerase activity could be detected in medium supernatants during treatment with any of these drugs, indicating that in vitro treatment did not severely affect the viability of metacestode tissue. Corresponding TEM analysis also revealed a dramatic shortening/retraction of microtriches as a hallmark of benzimidazole action, and as a consequence separation of the acellular laminated layer from the cellular germinal layer. Since TEM did not reveal any microtubule-based structures within Echinococcus microtriches, this effect cannot be explained by the previously described mechanism of action of benzimidazoles targeting β-tubulin, thus benzimidazoles must interact with additional targets that have not been yet identified. In addition, these results indicate the potential usefulness of fenbendazole for the chemotherapy of AE. Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  14. High prevalence of Taenia saginata taeniasis and status of Taenia solium cysticercosis in Bali, Indonesia, 2002-2004.

    Science.gov (United States)

    Wandra, T; Sutisna, P; Dharmawan, N S; Margono, S S; Sudewi, R; Suroso, T; Craig, P S; Ito, A

    2006-04-01

    An epidemiological survey of taeniasis/cysticercosis was carried out in one semi-urban and two urban villages in three districts of Bali, Indonesia in 2002 and 2004. In total, 398 local people from 247 families were diagnosed by anamnesis and clinical examinations, and 60 residents were suspected to be taeniasis carriers. Among 60 suspected carriers, 56 persons expelled a total of 61 taeniid adult worms after praziquantel treatment. From 398 residents, 252 stool samples were available for analysis of taeniid eggs, coproantigens or copro-DNA for identification of taeniid species, and 311 serum samples were available for detection of antibodies against Taenia solium cysticercosis. Taeniasis prevalences were highly variable among three villages (1.1-27.5%), and only one case of cysticercosis due to T. solium infection was detected. All expelled tapeworms were confirmed to be Taenia saginata by mtDNA analysis. There was no Taenia asiatica human case in Bali. Case control analysis of 106 families chosen at random from 179 families in 2004 and another 106 families from non-endemic areas revealed that risk factors of T. saginata taeniasis for families were: level of education (P<0.01); consumption of beef lawar (P<0.01); and the source of lawar (P<0.01).

  15. Molecular analyses reveal two geographic and genetic lineages for tapeworms, Taenia solium and Taenia saginata, from Ecuador using mitochondrial DNA.

    Science.gov (United States)

    Solano, Danilo; Navarro, Juan Carlos; León-Reyes, Antonio; Benítez-Ortiz, Washington; Rodríguez-Hidalgo, Richar

    2016-12-01

    Tapeworms Taenia solium and Taenia saginata are the causative agents of taeniasis/cysticercosis. These are diseases with high medical and veterinary importance due to their impact on public health and rural economy in tropical countries. The re-emergence of T. solium as a result of human migration, the economic burden affecting livestock industry, and the large variability of symptoms in several human cysticercosis, encourage studies on genetic diversity, and the identification of these parasites with molecular phylogenetic tools. Samples collected from the Ecuadorian provinces: Loja, Guayas, Manabí, Tungurahua (South), and Imbabura, Pichincha (North) from 2000 to 2012 were performed under Maximum Parsimony analyses and haplotype networks using partial sequences of mitochondrial DNA, cytochrome oxidase subunit I (COI) and NADH subunit I (NDI), from Genbank and own sequences of Taenia solium and Taenia saginata from Ecuador. Both species have shown reciprocal monophyly, which confirms its molecular taxonomic identity. The COI and NDI genes results suggest phylogenetic structure for both parasite species from south and north of Ecuador. In T. solium, both genes gene revealed greater geographic structure, whereas in T. saginata, the variability for both genes was low. In conclusion, COI haplotype networks of T. solium suggest two geographical events in the introduction of this species in Ecuador (African and Asian lineages) and occurring sympatric, probably through the most common routes of maritime trade between the XV-XIX centuries. Moreover, the evidence of two NDI geographical lineages in T. solium from the north (province of Imbabura) and the south (province of Loja) of Ecuador derivate from a common Indian ancestor open new approaches for studies on genetic populations and eco-epidemiology. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Acute intestinal obstruction due to taenia saginata infestation: a case report

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    Soleimani A

    2011-05-01

    Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 st1":*{behavior:url(#ieooui } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Infection with Taenia saginata or taeniasis is an uncommon parasitic infection in Iran with a prevalence rate of 2-3% and it is more seen in the northern parts of the country. Epigastric pain, nervousness, dizziness, nausea and loss of appetite may be the only presenting symptoms but secondary appendicitis, acute intestinal obstruction and necrosis of the pancreas are its serious and rare complications."n"nCase presentation : A 62-year old woman was admitted to Imam Khomeini Hospital with signs of acute abdomen. She had a past history of infection with hydatid cyst and its subsequent surgery, eight years ago. At the time of admission, she suffered from persistent abdominal pain and loss of appetite for two years. Despite having the epidemiological evidence of working along the banks of rivers contaminated with human sewage and working on farms fertilized with human waste and presence of signs hinting at the disease, parasitic infection had not been considered in its diagnosis."n"nConclusion: Although signs and symptoms of taeniasis are non-specific but a complete history, physical examination and detailed patient notes, especially by considering epidemiological factors, are

  17. Human Taeniasis and cysticercosis in Asia.

    Science.gov (United States)

    Ito, Akira; Nakao, Minoru; Wandra, Toni

    2003-12-06

    Human Taeniasis caused by the pork, Taenia solium, or beef, T saginata, tapeworm arises after eating pork or beef contaminated with metacestodes, the larval stage of these parasites. Taeniasis with T solium can lead to neurocysticercosis and threaten others by accidental ingestion of eggs released from asymptomatic Taeniasis patients. The 2003 World Health Assembly declared that T solium is of worldwide public-health importance, and that it is an eradicable parasitic disease worldwide. Adult taeniid tapeworms expelled from people in almost all Asian countries appeared to be T saginata (the so-called Asian Taenia), even though they ate pork. The organism is now named T asiatica, and has been found in Taiwan, Korea, China, Vietnam, and Indonesia. But it has been difficult to differentiate T saginata from beef and Asian Taenia from pork. Marshall Lightowlers and colleagues (Int J Parasitol 2003; 33: 1207-17) recently demonstrated that recombinant oncosphere vaccines against several taeniid cestodes, including T ovis, T saginata, T solium, and Echinococcus granulosus, are highly effective. Protection was almost 100%, in the laboratory and in the field. These researchers found several common features, including a predicted secretory signal sequence, and one or two copies of a fibronectin type III domain, each encoded by separate exons within the associated gene. WHERE NEXT? Molecular and immunological techniques, including vaccine research and development of animal models for differentiation of taeniid species in humans, have greatly advanced over the past decade. The clinical importance of infections by these taeniids, including T asiatica, in humans, and the potential for cysticercosis attributable to T asiatica in humans, needs further study.

  18. Proliferative peritoneal and pleural cestodiasis in a cat caused by metacestodes of Mesocestoides sp. anatomohistopathological findings and genetic identification

    Directory of Open Access Journals (Sweden)

    Eleni C.

    2007-03-01

    Full Text Available A 10-year-old female cat was brought to Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana for post-mortem examination. The animal used to live, together with 26 other cats, in the big terrace of an apartment at the 8th floor in Rome, and was always fed with industrial pet food. Anamnesis referred balance troubles, vomit and convulsions, during a couple of days, followed by sudden death. At necropsy, the cat presented mucoid rhinitis, purulent tracheitis, small areas of pneumonia, dark spots in the liver, catarrhal-hemorrhagic gastritis, fibrinous enteritis and meningeal hyperemia. Thoracic and abdominal cavities were completely invaded by hundreds of larval stages of cestodes. The same parasites were also included in nodules in pancreatic, lung and kidney parenchyma. Microscopic examination of parasites allowed their identification as larval stages (metacestodes of cestodes of the genus Mesocestoides. The molecular genotyping of the metacestodes indicates a close relationship with members of the genus Mesocestoides, although a significant variation was found with respect to the available sequences of other species of the genus.

  19. Recent situation of taeniasis in Mongolia (2002-2012).

    Science.gov (United States)

    Davaasuren, Anu; Dorjsuren, Temuulen; Yanagida, Tetsuya; Sako, Yasuhito; Nakaya, Kazuhiro; Davaajav, Abmed; Agvaandaram, Gurbadam; Enkhbat, Tsatsral; Gonchigoo, Battsetseg; Dulmaa, Nyamkhuu; Chuluunbaatar, Gantigmaa; Ito, Akira

    2014-04-01

    Epidemiological situation of taeniasis in Mongolia was assessed based on mitochondrial DNA identification of the parasite species. Multiplex PCR was used on a total of 194 proglottid specimens of Taenia species and copro-PCR and loop-mediated isothermal amplification (LAMP) assays were utilized for detection of copro-DNA of 37 fecal samples from taeniasis patients submitted to the Mongolian National Center for Communicable Diseases (NCCD) from 2002 to 2012. In addition, 4 out of 44 calcified cysts in beef kept in formalin since 2003 were evaluated for histopathological confirmation of cattle cysticercosis. All proglottid specimens and stool samples were confirmed to be Taenia saginata by multiplex PCR and by copro-PCR and LAMP, respectively. Cysts collected from cattle were morphologically confirmed to be metacestodes of Taenia species. T. saginata taeniasis was identified from almost all ages from a 2-year-old boy up to a 88-year-old woman and most prominently in 15-29 age group (37%, 74/198) followed by 30-44 age group (34.8%, 69/198 ) from 15 of Mongolia's 21 provinces, while cattle cysticerci were found from 12 provinces. The highest proportion of taeniasis patients was in Ulaanbaatar, the capital of Mongolia.

  20. An unexpected resident in the ileum detected during robot-assisted laparoscopic radical cystoprostatectomy and intracorporeal Studer pouch formation: Taenia saginata parasite.

    Science.gov (United States)

    Canda, Abdullah Erdem; Asil, Erem; Balbay, Mevlana Derya

    2011-02-01

    A case of moving ileal Taenia saginata parasites is presented with demonstrative images. We came across the parasites surprisingly while performing robot-assisted laparoscopic radical cystoprostatectomy with intracorporeal Studer pouch urinary diversion. We recommend stool sample evaluation in the preoperative period for possible presence of intestinal parasitic diseases, particularly in patients with bladder cancer who are admitted from areas with an increased incidence of intestinal parasitic diseases, before opening the bowel segments during surgery to perform radical cystectomy and urinary diversion.

  1. Oxfendazole as successful treatment of Taenia hydatigena metacestodes in naturally infected pigs

    Institute of Scientific and Technical Information of China (English)

    Luis Antonio Gomez-Puerta; Armando Emiliano Gonzalez; Cesar Gavidia; Viterbo Ayvar; Hector Hugo Garcia; Maria Teresa Lopez-Urbina

    2015-01-01

    The efficacy of oxfendazole (OFZ) on Taenia hydatigena metacestodes, also called Cysticercus tenuicollis (C. tenuicollis), was studied in 648 raising pigs. This study was performed in Tumbes Department in Peru, an endemic area for cysticercosis. Pigs were randomized in two groups;untreated group (n=142) did not receive any treatment and treated group (n=506) received OFZ treatment at a single dose of 30 mg/kg body weight. Six months after treatment, the pigs were necropsied. The prevalence of infection by C. tenuicollis among the pigs was 27.5%(39/142) and 2.0%(10/506) in untreated and treated groups, respectively. Untreated group was infested only with viable cysts, whereas treated group had no viable cysts. All the cysts found in treated group presented degeneration, with a thick membrane, and they contained milky fluid and fibrous tissue. A single dose of OFZ was effective against C. tenuicollis, thus providing an alternative drug for controlling this parasite in pigs.

  2. Serological diagnosis of Taenia solium in pigs: No measurable circulating antigens and antibody response following exposure to Taenia saginata oncospheres.

    Science.gov (United States)

    Dorny, P; Dermauw, V; Van Hul, A; Trevisan, C; Gabriël, S

    2017-10-15

    Taenia solium taeniasis/cysticercosis is a zoonosis included in the WHO's list of neglected tropical diseases. Accurate diagnostic tools for humans and pigs are needed to monitor intervention outcomes. Currently used diagnostic tools for porcine cysticercosis all have drawbacks. Serological tests are mainly confronted with problems of specificity. More specifically, circulating antigen detecting tests cross-react with Taenia hydatigena and the possibility of transient antigens as a result of aborted infections is suspected. Furthermore, the hypothesis has been raised that hatched ingested eggs of other Taenia species may lead to a transient antibody response or to the presence of circulating antigen detectable by serological tests used for porcine cysticercosis. Here we describe the results of a study that consisted of oral administration of Taenia saginata eggs to five piglets followed by serological testing during five weeks and necropsy aiming at studying possible cross reactions in serological tests used for porcine cysticercosis. The infectivity of the eggs was verified by in vitro hatching and by experimental infection of a calf. One piglet developed acute respiratory disease and died on day 6 post infection. The remaining four piglets did not show any clinical signs until euthanasia. None of the serum samples from four piglets collected between days 0 and 35 post infection gave a positive reaction in the B158/B60 Ag-ELISA and in a commercial Western blot for antibody detection. In conclusion, this study showed that experimental exposure of four pigs to T. saginata eggs did not result in positive serologies for T. solium. These results may help interpreting serological results in monitoring of T. solium control programmes. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. [Urban alternate host cycle of Taenia saginata in Wittstock County, district of Potsdam (East Germany)].

    Science.gov (United States)

    Engelbrecht, H; Mentzel, U

    1984-08-01

    The urban alternate host cycle of Taenia saginata in the county of Wittstock, district of Potsdam (GDR). Investigations of the epidemiology of bovine cysticercosis and human taeniasis were carried out in the county of Wittstock from 1979 till 1980. The analysis was based on the examination of human faeces, sewage and sewage sludge, the registration of infested humans, anticestodica consumption, number of infested cattle, areas for squirting out sewage, fodder supply areas and location of cattle. These data were mapped. There is a direct connection between the prevalence of Cysticercus bovis in more than 50 per cent of the infested cattle of the county of Wittstock and the squirting out of the sewage of the county town. 21 to 27 million Taenia eggs per day get into the waste-water purification plant. The proportion of infested humans and infested cattle is 1: 250 or 1: 290, respectively. The prevalence of adult tapeworms is 0.008-0.18%, and that of cysticerci is 20%.

  4. Impairment of the chemical defence of the beetle, Tenebrio molitor, by metacestodes (cysticeroids) of the tapeworm, Hymenolepis diminuta.

    Science.gov (United States)

    Blankespoor, C L; Pappas, P W; Eisner, T

    1997-07-01

    The defensive glands of beetles, Tenebrio molitor, infected with metacestodes (cysticercoids) of Hymenolepis diminuta are everted less frequently upon stimulation, and contain less toluquinone (methylbenzoquinone) and m-cresol, than glands of uninfected controls. These differences, as shown in predation trials with wild rats, increase the likelihood that both cysticercoids and beetles will be ingested by the tapeworm's definitive host. This is the first documented case of a parasite inhibiting the chemical defence of an intermediate host, and one of only a few reports of parasite-induced manipulation of host biology supported by empirical evidence implicating facilitated parasite transmission between host species.

  5. Purifying Nanomaterials

    Science.gov (United States)

    Hung, Ching-Cheh (Inventor); Hurst, Janet (Inventor)

    2014-01-01

    A method of purifying a nanomaterial and the resultant purified nanomaterial in which a salt, such as ferric chloride, at or near its liquid phase temperature, is used to penetrate and wet the internal surfaces of a nanomaterial to dissolve impurities that may be present, for example, from processes used in the manufacture of the nanomaterial.

  6. Sequence analysis and molecular characterization of Wnt4 gene in metacestodes of Taenia solium.

    Science.gov (United States)

    Hou, Junling; Luo, Xuenong; Wang, Shuai; Yin, Cai; Zhang, Shaohua; Zhu, Xueliang; Dou, Yongxi; Cai, Xuepeng

    2014-04-01

    Wnt proteins are a family of secreted glycoproteins that are evolutionarily conserved and considered to be involved in extensive developmental processes in metazoan organisms. The characterization of wnt genes may improve understanding the parasite's development. In the present study, a wnt4 gene encoding 491amino acids was amplified from cDNA of metacestodes of Taenia solium using reverse transcription PCR (RT-PCR). Bioinformatics tools were used for sequence analysis. The conserved domain of the wnt gene family was predicted. The expression profile of Wnt4 was investigated using real-time PCR. Wnt4 expression was found to be dramatically increased in scolex evaginated cysticerci when compared to invaginated cysticerci. In situ hybridization showed that wnt4 gene was distributed in the posterior end of the worm along the primary body axis in evaginated cysticerci. These findings indicated that wnt4 may take part in the process of cysticerci evagination and play a role in scolex/bladder development of cysticerci of T. solium.

  7. High-throughput characterization of Echinococcus spp. metacestode miRNomes.

    Science.gov (United States)

    Cucher, Marcela; Macchiaroli, Natalia; Kamenetzky, Laura; Maldonado, Lucas; Brehm, Klaus; Rosenzvit, Mara Cecilia

    2015-03-01

    Echinococcosis is a worldwide zoonosis of great public health concern, considered a neglected disease by the World Health Organisation. The cestode parasites Echinococcus granulosus sensu lato (s. l.) and Echinococcus multilocularis are the main aetiological agents. In the intermediate host, these parasites display particular developmental traits that lead to different patterns of disease progression. In an attempt to understand the causes of these differences, we focused on the analysis of microRNAs (miRNAs), small non-coding regulatory RNAs with major roles in development of animals and plants. In this work, we analysed the small RNA expression pattern of the metacestode, the stage of sanitary relevance, and provide a detailed description of Echinococcus miRNAs. Using high-throughput small RNA sequencing, we believe that we have carried out the first experimental identification of miRNAs in E. multilocularis and have expanded the Echinococcus miRNA catalogue to 38 miRNA genes, including one miRNA only present in E. granulosus s. l. Our findings show that although both species share the top five highest expressed miRNAs, 13 are differentially expressed, which could be related to developmental differences. We also provide evidence that uridylation is the main miRNA processing mechanism in Echinococcus spp. These results provide detailed information on Echinococcus miRNAs, which is the first step in understanding their role in parasite biology and disease establishment and/or progression, and their future potential use as drug or diagnostic targets. Copyright © 2015 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  8. First report of metacestodes of Cyclustera ralli (Underwood and Dronen, 1986) (Cestoda: Cyclophyllidea) from the fish intermediate host, Fundulus heteroclitus (Linnaeus, 1766) (Cyprinodontiformes: Fundulidae) from Virginia, U.S.A

    Czech Academy of Sciences Publication Activity Database

    Scholz, Tomáš; Harris, C. E.

    2006-01-01

    Roč. 73, č. 1 (2006), s. 130-133 ISSN 1525-2647 R&D Projects: GA ČR GA524/04/0342; GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : metacestodes * fish * morphology Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 0.659, year: 2006

  9. Does interspecific competition have a moderating effect on Taenia solium transmission dynamics in Southeast Asia?

    Science.gov (United States)

    Conlan, James V; Vongxay, Khamphouth; Fenwick, Stanley; Blacksell, Stuart D; Thompson, R C Andrew

    2009-09-01

    It is well understood that sociocultural practices strongly influence Taenia solium transmission; however, the extent to which interspecific parasite competition moderates Taenia transmission has yet to be determined. This is certainly the case in Southeast Asia where T. solium faces competition in both the definitive host (people) and the intermediate host (pigs). In people, adult worms of T. solium, T. saginata and T. asiatica compete through density-dependent crowding mechanisms. In pigs, metacestodes of T. solium, T. hydatigena and T. asiatica compete through density-dependent immune-mediated interactions. Here, we describe the biological and epidemiological implications of Taenia competition and propose that interspecific competition has a moderating effect on the transmission dynamics of T. solium in the region. Furthermore, we argue that this competitive ecological scenario should be considered in future research and surveillance activities examining T. solium cysticercosis and taeniasis in Southeast Asia.

  10. Case Record of a Teaching Hospital in Karaj; A 35-Year Old Man With Taenia saginata Infection Treated With Niclosamide

    Directory of Open Access Journals (Sweden)

    Aliehsan Heidari

    2016-11-01

    Full Text Available Taeniasaginata can cause severe health and economic problems particularly in endemic areas. The disease cause by this cestode is related to poor sanitary conditions, inadequate hygiene, open defecation, inadequately cooked beef and poverty. A 35 years man found yellowish white tapeworm proglottids moving in his feces and consulted to the Department of Emergency, ShahidRajaei hospital, Karaj, Iran. He complained of lower abdominal discomfort, anal itching and moving something in the stomach. He was given wrong prescription. The patients had the history of eating undercooked beef. We report one case of T. saginata infection based on adult tapeworm recovery from the patients. The specific identification of the worm was based on based on standard procedures. Three months after expelling the tapeworm, the man felt better and returned to his normal life.

  11. Structural and binding properties of two paralogous fatty acid binding proteins of Taenia solium metacestode.

    Directory of Open Access Journals (Sweden)

    Seon-Hee Kim

    Full Text Available BACKGROUND: Fatty acid (FA binding proteins (FABPs of helminths are implicated in acquisition and utilization of host-derived hydrophobic substances, as well as in signaling and cellular interactions. We previously demonstrated that secretory hydrophobic ligand binding proteins (HLBPs of Taenia solium metacestode (TsM, a causative agent of neurocysticercosis (NC, shuttle FAs in the surrounding host tissues and inwardly transport the FAs across the parasite syncytial membrane. However, the protein molecules responsible for the intracellular trafficking and assimilation of FAs have remained elusive. METHODOLOGY/PRINCIPAL FINDINGS: We isolated two novel TsMFABP genes (TsMFABP1 and TsMFABP2, which encoded 133- and 136-amino acid polypeptides with predicted molecular masses of 14.3 and 14.8 kDa, respectively. They shared 45% sequence identity with each other and 15-95% with other related-members. Homology modeling demonstrated a characteristic β-barrel composed of 10 anti-parallel β-strands and two α-helices. TsMFABP2 harbored two additional loops between β-strands two and three, and β-strands six and seven, respectively. TsMFABP1 was secreted into cyst fluid and surrounding environments, whereas TsMFABP2 was intracellularly confined. Partially purified native proteins migrated to 15 kDa with different isoelectric points of 9.2 (TsMFABP1 and 8.4 (TsMFABP2. Both native and recombinant proteins bound to 11-([5-dimethylaminonaphthalene-1-sulfonyl]aminoundecannoic acid, dansyl-DL-α-amino-caprylic acid, cis-parinaric acid and retinol, which were competitively inhibited by oleic acid. TsMFABP1 exhibited high affinity toward FA analogs. TsMFABPs showed weak binding activity to retinol, but TsMFABP2 showed relatively high affinity. Isolation of two distinct genes from an individual genome strongly suggested their paralogous nature. Abundant expression of TsMFABP1 and TsMFABP2 in the canal region of worm matched well with the histological distributions

  12. Endurance Pump Test with MIL-PRF-83282 Hydraulic Fluid, Purified with Malabar Purifier

    National Research Council Canada - National Science Library

    Sharma, Shashi

    2004-01-01

    .... Endurance aircraft hydraulic pump tests under carefully controlled conditions were previously conducted using hydraulic fluid purified with a rotating-disk and vacuum type purifier, the portable...

  13. Handbook of purified gases

    CERN Document Server

    Schoen, Helmut

    2015-01-01

    Technical gases are used in almost every field of industry, science and medicine and also as a means of control by government authorities and institutions and are regarded as indispensable means of assistance. In this complete handbook of purified gases the physical foundations of purified gases and mixtures as well as their manufacturing, purification, analysis, storage, handling and transport are presented in a comprehensive way. This important reference work is accompanied with a large number of Data Sheets dedicated to the most important purified gases.  

  14. Histological and ultrastructural localization of antigen B in the metacestode of Taenia solium

    International Nuclear Information System (INIS)

    Laclette, J.P.; Merchant, M.T.; Willms, K.

    1987-01-01

    The morphological localization of antigen B (AgB) in the tissues of the Taenia solium metacestode was studied by immunological and biochemical methods. Indirect immunofluorescence carried out on vibratome sections showed that AgB is widely distributed throughout the tissue. A more intense fluorescence was observed in the tegumentary cytons of the bladder wall and in the lumen of the spiral canal of the invaginated scolex. Ultrastructural analysis of larvae washed in PBS after dissection from meat and then incubated with rabbit antibodies against AgB, followed by peroxidase-labeled goat anti-rabbit IgG, did not exhibit electron-dense material on the external surface. Larvae fixed in glutaraldehyde immediately after dissection and exposed to the immunoperoxidase reagents did exhibit electron-dense material on microtriches, indicating that AgB is only loosely bound to the external surface. Crude extracts of surface-radioiodinated cysticerci analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) contained no labeled proteins with the molecular weight of AgB. Autoradiography of the immunoelectrophoretograms in which the crude extract was confronted with antibodies to AgB demonstrated that this antigen was not labeled, and therefore is not exposed on the tegumentary surface. The results suggest that AgB is synthesized by the tegumentary cytons of the parasite and secreted through the tegumental membrane into the host tissues and the lumen of the spiral canal

  15. Histological and ultrastructural localization of antigen B in the metacestode of Taenia solium

    Energy Technology Data Exchange (ETDEWEB)

    Laclette, J.P.; Merchant, M.T.; Willms, K.

    1987-02-01

    The morphological localization of antigen B (AgB) in the tissues of the Taenia solium metacestode was studied by immunological and biochemical methods. Indirect immunofluorescence carried out on vibratome sections showed that AgB is widely distributed throughout the tissue. A more intense fluorescence was observed in the tegumentary cytons of the bladder wall and in the lumen of the spiral canal of the invaginated scolex. Ultrastructural analysis of larvae washed in PBS after dissection from meat and then incubated with rabbit antibodies against AgB, followed by peroxidase-labeled goat anti-rabbit IgG, did not exhibit electron-dense material on the external surface. Larvae fixed in glutaraldehyde immediately after dissection and exposed to the immunoperoxidase reagents did exhibit electron-dense material on microtriches, indicating that AgB is only loosely bound to the external surface. Crude extracts of surface-radioiodinated cysticerci analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) contained no labeled proteins with the molecular weight of AgB. Autoradiography of the immunoelectrophoretograms in which the crude extract was confronted with antibodies to AgB demonstrated that this antigen was not labeled, and therefore is not exposed on the tegumentary surface. The results suggest that AgB is synthesized by the tegumentary cytons of the parasite and secreted through the tegumental membrane into the host tissues and the lumen of the spiral canal.

  16. Antigenic fractions from Taenia crassiceps metacestodes obtained by hydrophobicity for the immunodiagnosis of active and inactive forms of neurocysticercosis in human cerebrospinal fluid samples.

    Science.gov (United States)

    da Silva, Gabriela B; Nunes, Daniela S; de Sousa, José Eduardo N; Gonçalves-Pires, Maria do R F; Levenhagen, Marcelo A; Costa-Cruz, Julia M

    2017-04-01

    This study aimed to evaluate the total extract of Taenia crassiceps metacestodes (TC) and its antigenic fractions obtained by Triton X-114 fractionation techniques, such as detergent (DC) and aqueous (AC), in the immunodiagnosis of human neurocysticercosis (NCC). Cerebrospinal fluid samples were divided into two groups: Group 1 (n=40), which was further divided into active (n=20) and inactive (n=20) NCC, and Group 2 (control group), which comprised 39 CSF samples from patients who had another neurological disorder, were suffering from other infectious diseases of the brain or had other parasitic infections. The total extracts and antigenic fractions were tested by enzyme-linked immunosorbent assay (ELISA) to detect human IgG anti-Taenia solium. T. crassiceps fractions (DC and AC) showed the same value of sensitivity (Se), 100%, for active and inactive NCC and a specificity (Sp) of 97.4%. The DS fraction obtained from T. solium showed 100% Se for active NCC, 95% Se for inactive NCC and a 92.3% Sp. The AS fraction obtained from T. solium showed 100% Se for both active and inactive NCC and a 94.9% Sp. There was a positive correlation between the total saline extract of T. crassiceps (TC) and T. solium (TS) and their fractions (DC, AC, DS and AS). Positive predictive value, negative predictive value, diagnostic efficiency and Youden index were calculated. In conclusion, these results demonstrated that detergent and aqueous fractions obtained from T. crassiceps metacestodes are important sources of specific antigens and are efficient for immunodiagnosis of active and inactive NCC. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

  17. Identification and functional characterization of alpha-enolase from Taenia pisiformis metacestode.

    Science.gov (United States)

    Zhang, Shaohua; Guo, Aijiang; Zhu, Xueliang; You, Yanan; Hou, Junling; Wang, Qiuxia; Luo, Xuenong; Cai, Xuepeng

    2015-04-01

    Enolase belongs to glycolytic enzymes with moonlighting functions. The role of enolase in Taenia species is still poorly understood. In this study, the full length of cDNA encoding for Taenia pisiformis alpha-enolase (Tpeno) was cloned from larval parasites and soluble recombinant Tpeno protein (rTpeno) was produced. Western blot indicated that both rTpeno and the native protein in excretion-secretion antigens from the larvae were recognized by anti-rTpeno monoclonal antibodies (MAbs). The primary structure of Tpeno showed the presence of a highly conserved catalytic site for substrate binding and an enolase signature motif. rTpeno enzymatic activities of catalyzing the reversible dehydration of 2-phosphoglycerate (2-PGA) to phosphoenolpyruvate (PEP) and vice versa were shown to be 30.71 ± 2.15 U/mg (2-PGA to PEP) and 11.29 ± 2.38 U/mg (PEP to 2-PGA), respectively. Far-Western blotting showed that rTpeno could bind to plasminogen, however its binding ability was inhibited by ϵ-aminocaproic acid (ϵACA) in a competitive ELISA test. Plasminogen activation assay showed that plasminogen bound to rTpeno could be converted into active plasmin using host-derived activators. Immunohistochemistry and immunofluorescence indicated that Tpeno was distributed in the bladder wall of the metacestode and the periphery of calcareous corpuscles. In addition, a vaccine trial showed that the enzyme could produce a 36.4% protection rate in vaccinated rabbits against experimental challenges from T. pisiformis eggs. These results suggest that Tpeno with multiple functions may play significant roles in the migration, growth, development and adaptation of T. pisiformis for survival in the host environment. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Purifying hydrocarbons

    Energy Technology Data Exchange (ETDEWEB)

    Dunstan, A E

    1918-06-03

    Ligroin, kerosene, and other distillates from petroleum and shale oil, are purified by treatment with a solution of a hypochlorite containing an excess of alkali. The hydrocarbon may be poured into brine, the mixture stirred, and an electric current passed through. Heat may be applied.

  19. Pentacene field-effect transistors by in situ and real time electrical characterization: Comparison between purified and non-purified thin films

    International Nuclear Information System (INIS)

    Liu, Shun-Wei; Wen, Je-Min; Lee, Chih-Chien; Su, Wei-Cheng; Wang, Wei-Lun; Chen, Ho-Chien; Lin, Chun-Feng

    2013-01-01

    We present an electrical characterization of the organic field-effect transistor with purified and non-purified pentacene by using in situ and real time measurements. The field-effect phenomenon was observed at the thickness of 1.5 nm (approximately one monolayer of pentacene) for purified pentacene, as compared to 3.0 nm for the non-purified counterpart. Moreover, the hole mobility is improved from 0.13 to 0.23 cm 2 /V s after the sublimation process to purify the pentacene. With atomic force microscopic measurements, the purified pentacene thin film exhibits a larger grain size and film coverage, resulting in better crystallinity of the thin film structure due to the absence of the impurities. This is further confirmed by X-ray diffraction patterns, which show higher intensities for the purified pentacene. - Highlights: • We present in-situ characterization for pentacene field-effect transistors. • The hole mobility is improved after the sublimation process to purify the pentacene. • Purified pentacene thin film exhibits a larger grain size and film coverage. • Hole mobility of pentacene is improved from 0.13 to 0.23 cm 2 /V s. • The discontinuity of grain boundary may cause the shift of threshold voltage

  20. Pentacene field-effect transistors by in situ and real time electrical characterization: Comparison between purified and non-purified thin films

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Shun-Wei, E-mail: swliu@mail.mcut.edu.tw [Department of Electronic Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan, ROC (China); Wen, Je-Min; Lee, Chih-Chien; Su, Wei-Cheng; Wang, Wei-Lun; Chen, Ho-Chien [Department of Electronic Engineering, National Taiwan University of Science and Technology, Taipei, 10607 Taiwan, ROC (China); Lin, Chun-Feng [Department of Electronic Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan, ROC (China)

    2013-05-01

    We present an electrical characterization of the organic field-effect transistor with purified and non-purified pentacene by using in situ and real time measurements. The field-effect phenomenon was observed at the thickness of 1.5 nm (approximately one monolayer of pentacene) for purified pentacene, as compared to 3.0 nm for the non-purified counterpart. Moreover, the hole mobility is improved from 0.13 to 0.23 cm{sup 2}/V s after the sublimation process to purify the pentacene. With atomic force microscopic measurements, the purified pentacene thin film exhibits a larger grain size and film coverage, resulting in better crystallinity of the thin film structure due to the absence of the impurities. This is further confirmed by X-ray diffraction patterns, which show higher intensities for the purified pentacene. - Highlights: • We present in-situ characterization for pentacene field-effect transistors. • The hole mobility is improved after the sublimation process to purify the pentacene. • Purified pentacene thin film exhibits a larger grain size and film coverage. • Hole mobility of pentacene is improved from 0.13 to 0.23 cm{sup 2}/V s. • The discontinuity of grain boundary may cause the shift of threshold voltage.

  1. Methods for Purifying Enzymes for Mycoremediation

    Science.gov (United States)

    Cullings, Kenneth W. (Inventor); DeSimone, Julia C. (Inventor); Paavola, Chad D. (Inventor)

    2014-01-01

    A process for purifying laccase from an ectomycorrhizal fruiting body is disclosed. The process includes steps of homogenization, sonication, centrifugation, filtration, affinity chromatography, ion exchange chromatography, and gel filtration. Purified laccase can also be separated into isomers.

  2. Hydrogen purifier module with membrane support

    Science.gov (United States)

    A hydrogen purifier utilizing a hydrogen-permeable membrane to purify hydrogen from mixed gases containing hydrogen is disclosed. Improved mechanical support for the permeable membrane is described, enabling forward or reverse differential pressurization of the membrane, which further stabilizes the membrane from wrinkling upon hydrogen uptake.

    2012-07-24

    A hydrogen purifier utilizing a hydrogen-permeable membrane to purify hydrogen from mixed gases containing hydrogen is disclosed. Improved mechanical support for the permeable membrane is described, enabling forward or reverse differential pressurization of the membrane, which further stabilizes the membrane from wrinkling upon hydrogen uptake.

  3. 21 CFR 880.6710 - Medical ultraviolet water purifier.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Medical ultraviolet water purifier. 880.6710... Miscellaneous Devices § 880.6710 Medical ultraviolet water purifier. (a) Identification. A medical ultraviolet water purifier is a device intended for medical purposes that is used to destroy bacteria in water by...

  4. Fatal infection with Taenia martis metacestodes in a ring-tailed lemur (Lemur catta) living in an Italian zoological garden.

    Science.gov (United States)

    De Liberato, Claudio; Berrilli, Federica; Meoli, Roberta; Friedrich, Klaus G; Di Cerbo, Pilar; Cocumelli, Cristiano; Eleni, Claudia

    2014-10-01

    A case of fatal infection caused by larval forms of Taenia martis in a ring-tailed lemur (Lemur catta) living in the Rome zoological garden is described. The animal, living in a semi-natural pen with other 15 conspecific individuals and being fed with fresh fruit and vegetables, yoghurt and eggs, was transported to the Istituto Zooprofilattico of Rome for post-mortem examination. The anamnesis included, ten days before the death, apathy, lack of appetite, abdominal distension and diarrhoea. A severe exudative fibrinous-purulent peritonitis with numerous adhesions between the abdominal wall and the bowel loops was detected. After intestine removal, two free and viable, 4 cm long, whitish, leaf-like parasitic forms were pinpointed. Macroscopic examination of the two parasites allowed their identification as larval stages of cestodes, identified via molecular analysis as T. martis metacestodes. This report represents the first record of T. martis infection in the host species and in a zoological garden and for the pathological relevance of the infection. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  5. Anti-Taenia solium metacestode IgG antibodies in serum samples from inhabitants of a central-western region of Brazil

    Directory of Open Access Journals (Sweden)

    Oliveira Heliana B. de

    2006-01-01

    Full Text Available A total of 354 serum samples from inhabitants who frequent the Clinical Laboratory in Catalão, Goiás, in the central-western region of Brazil, were collected from June to August, 2002. The samples were evaluated by indirect immunofluorescence antibody tests and an enzyme linked immunosorbent assay in order to detect anti-Taenia solium metacestode IgG antibodies. Reactive and inconclusive samples were tested by Western blotting (WB. Considering WB as a confirmation, the frequency of antibodies in the serum samples of the above population was 11.3% (CI 5.09 - 17.51. The immunodominant bands most frequently recognized in WB were 64-68 kDa (97.5% and 47-52 kDa (80%. The percentage of seropositivity to cysticercosis was significantly higher for individuals residing in areas without sewage systems (p < 0.0001. In conclusion, the results indicate a probable endemic situation of cysticercosis in this population. These results reinforce the urgent need for control and prevention measures to be taken by the local public health services.

  6. Respirators: Air Purifying, Self-Study, Course 40723

    Energy Technology Data Exchange (ETDEWEB)

    Chochoms, Michael [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-12-21

    Respirators: Air Purifying Self-Study (COURSE 40723) is designed for Los Alamos National Laboratory (LANL) workers, support services subcontractors, and other LANL subcontractors who work under the LANL Respiratory Protection Program (RPP). This course also meets the air-purifying respirators (APRs) retraining requirement.

  7. Methods for purifying carbon materials

    Science.gov (United States)

    Dailly, Anne [Pasadena, CA; Ahn, Channing [Pasadena, CA; Yazami, Rachid [Los Angeles, CA; Fultz, Brent T [Pasadena, CA

    2009-05-26

    Methods of purifying samples are provided that are capable of removing carbonaceous and noncarbonaceous impurities from a sample containing a carbon material having a selected structure. Purification methods are provided for removing residual metal catalyst particles enclosed in multilayer carbonaceous impurities in samples generate by catalytic synthesis methods. Purification methods are provided wherein carbonaceous impurities in a sample are at least partially exfoliated, thereby facilitating subsequent removal of carbonaceous and noncarbonaceous impurities from the sample. Methods of purifying carbon nanotube-containing samples are provided wherein an intercalant is added to the sample and subsequently reacted with an exfoliation initiator to achieve exfoliation of carbonaceous impurities.

  8. Study of hot corrosion of flakes of non purified graphite and of purified graphite

    International Nuclear Information System (INIS)

    Boule, Michel

    1967-01-01

    The author reports the study of hot corrosion of the Ticonderoga graphite. He reports the study of the defects of graphite flakes (structure defects due to impurities), the dosing of these impurities, and then their removal by purification. Flakes have then been oxidised by means of a specially designed apparatus. Based on photographs taken by optical and electronic microscopy, the author compares the oxidation features obtained in dry air and in humid air, between purified and non purified flakes. He also reports the study of the evolution of oxidation with respect to the initial rate of impurities, and the study of the evolution of oxidation features in humid air during oxidation. All these comparisons are made while taking the oxidation rate into account [fr

  9. Light aging of reactive fuels purified by various methods

    Energy Technology Data Exchange (ETDEWEB)

    Khodzhaeva, M G; Burtyshev, N Ya; Molodozhenyuk, T B; Ryabovda, N D

    1976-01-01

    A study of the effect of uv-radiation on aging of Fergana fuel TS-1 has been extended to the uv-effect on alkali-purified fuels (e.g., Krasnovodsk, Omsk, and Orsk TS-1), on hydro-purified (Syzran T-8, Syzran T-7, and Novokuybyshev T-7) and on adsorption-purified Fergana TS-1. The PRK-4 lamp was employed. Aging criteria were formation of insoluble gums, soluble gums separable on silicagel, acidity, and optical density. Fuels purified in the same manner aged practically identically; after 6 months storage the greatest gum formation was seen in the fuels Orsk TS-1 and Syzran T-8. 3 references, 1 figure, 1 table.

  10. Isoforms of purified methyltransferase from human blood platelets ...

    African Journals Online (AJOL)

    ... purification from normal human blood platelets have not been investigated, hence, the aim of this study was to purify, characterise the enzyme from human blood platelets and determine its possible role in phospholipid transmethylation. The plasma membranes were purified by velocity and sucrose gradient centrifugation ...

  11. Tapeworm infection - beef or pork

    Science.gov (United States)

    Teniasis; Pork tapeworm; Beef tapeworm; Tapeworm; Taenia saginata; Taenia solium; Taeniasis ... undercooked meat of infected animals. Cattle usually carry Taenia saginata ( T saginata ). Pigs carry Taenia solium (T ...

  12. Reproducible in vitro regeneration system for purifying sugarcane ...

    African Journals Online (AJOL)

    This procedure may be considered as one of the best ever published report on regeneration from in vitro grown plants to purify clones without subjecting the plants to field conditions and harvesting the mature cane. This technique was used to purify transgenic sugarcane plants carrying Bacillus thuringiensis gene.

  13. Purifying hydrocarbon oils

    Energy Technology Data Exchange (ETDEWEB)

    Rostin, H

    1938-08-11

    A process is described for continuously purifying hydrocarbon oils consisting in conducting the vapors of the same at a temperature of 300 to 400/sup 0/C over the oelitic ore minette together with reducing gases in presence of steam the proportion of the reducing gases and steam being such that the sulfur of the hydrocarbons escapes from the reaction chamber in the form of sulfuretted hydrogen without permanent sulfide of iron being formed.

  14. Taenia solium metacestode preparation in rural areas of sub-Saharan Africa: a source for diagnosis and research on cysticercosis.

    Science.gov (United States)

    Schmidt, V; Sikasunge, C S; Odongo-Aginya, E; Simukoko, C; Mwanjali, G; Alarakol, S; Ovuga, E; Matuja, W; Kihamia, C; Löscher, T; Winkler, A S; Bretzel, G

    2015-03-01

    Taenia solium metacestodes/cysts obtained from pig carcasses constitute a primary source for diagnostic tools used for the detection of human cysticercosis. Data on T. solium cyst preparation in Africa is still scarce but required to establish independent reference laboratories. The aim of the present study is a) to present the likely yield of T. solium cyst material by the use of two different preparation methods in the field and b) to investigate its suitability for immunodiagnosis of human cysticercosis. In Zambia, Uganda and Tanzania 670 pigs were screened for T. solium infection. Cysts were prepared by 'shaking method' and 'washing method'. Generated crude antigens were applied in a standard western blot assay. 46 out of 670 pigs (6.9%) were found positive for T. solium (Zambia: 12/367, 3.3%; Uganda: 11/217, 5.1%; Tanzania 23/86, 26.7%). Mean values of 77.7 ml whole cysts, 61.8 ml scolices/membranes and 10.9 ml cyst fluid were obtained per pig. Suitability of collected material for the use as crude antigen and molecular diagnostic techniques was demonstrated. This study clearly shows that T. solium cyst preparation in African settings by simple field methods constitutes an effective way to obtain high quality material as source for diagnostic tools and research purposes.

  15. Method and device for feeding purified water to a pressure vessel

    International Nuclear Information System (INIS)

    Hirato, Miharu.

    1982-01-01

    Purpose: To prevent thermal wear at the junction of feedwater pipes and purified water pipes, as well as maintain the function of the purified water feeding system by stopping the introduction of purified water to the heated water feeding system and introducing purified water to the recycling water system upon transient operation or start-up. Constitution: Since a feedwater heater does not function well during transient operation or upon start-up, the temperature of heated water flowing through the feedwater pipe is reduced to produce a temperature difference relative to the set temperature for the purified water feeding system. The temperature difference is detected by a temperature sensor and, when it arrives at a predetermined difference, an operation valve is switched to interrupt the feed of the purified water to the heated water feeding system and it is sent to a water recycling system. Then, the purified water is sent from the water recycling system by way of the discharge portion to the inside of a pressure vessel. Thus, since only the heated water flows to the junction between the cleaned water pipes and the heated water pipes, neither shocks are generated nor the performance of the purified water feeding system is impaired. (Moriyama, K.)

  16. Assay of partially purified glutamate dehydrogenase isolated from ...

    African Journals Online (AJOL)

    Glutamate dehydrogenase (E C 1.4.1.1) isolated from the seeds of asparagus beans was partially purified to a factor of 22 by dialysis after fractional precipitation with solid ammonium sulphate at 40 and 60% saturation. A specific activity of 11.78μmol min-1 mg-1 protein was calculated for the partially purified enzyme when ...

  17. Partially purified polygalacturonase from Aspergillus niger (SA6 ...

    African Journals Online (AJOL)

    Polygalacturonase (PG) was isolated from Aspergillus niger (A. niger) (SA6), partially purified and characterized. The PG showed two bands on SDS-PAGE suggesting an “endo and exo PG with apparent molecular weights of 35 and 40 KDa, respectively. It was purified 9-fold with a yield of 0.18% and specific activity of 246 ...

  18. Efecto de algunos agentes físicos y químicos sobre el metacéstodo de Taenia solium presente en carne adobada y chorizo Effects of some chemical and physical agents on the metacestode Taenia solium in spicy meat and sausage

    Directory of Open Access Journals (Sweden)

    Ma Isabel Rivera-Guerrero

    2004-10-01

    Full Text Available OBJETIVO: Evaluar el efecto de diferentes temperaturas y tiempos, así como de algunos condimentos sobre la viabilidad de metacéstodos de Taenia solium en chorizo y carne adobada. MATERIAL Y MÉTODOS: Este trabajo se llevó a cabo en la Universidad Autónoma de Guerrero, en 1999. En la comunidad de Atzacoaloya, en el municipio de Chilapa de Alvarez, Guerrero, se compró carne de cerdo infectada, con la que se preparó carne adobada y chorizo; sólo se empleó aquélla en la cual se comprobó la viabilidad de los metacéstodos. Los productos obtenidos fueron sometidos a: a temperatura ambiente durante 12 a 100 horas; b temperaturas de -10 a 37 ºC por 24 horas, y c ebullición (97 ºC de 1 a 15 minutos. Para determinar el efecto de los condimentos se prepararon lotes con el doble de ingredientes de cada uno. Todas las evaluaciones se realizaron y evaluaron con tres repeticiones. Se establecieron diferencias de proporciones mediante c². RESULTADOS: A temperatura ambiente la menor evaginación fue a las 100 horas para ambos productos (pOBJECTIVE: To assess the effect of different cooking times and temperatures, as well as of some seasonings, on the viability of Taenia solium metacestodes in spicy meat and hot sausage. MATERIAL AND METHODS: This study was conducted by the Universidad Autónoma de Guerrero (Guerrero State Autonomous University, Mexico in 1999. Infected pork meat was bought in the community of Azacoaloya, in the municipality of Chilapa de Alvarez, Guerrero State. It was used to prepare spicy meat (adobada and hot sausage (chorizo. Only the meat in which metacestode viability was proven was used. The products obtained underwent a room temperature for 12 to 100 hours; b temperatures of -10 to 37ºC for 24 hours; c boiling (97ºC from 1 to 15 minutes. To determine the effect of the seasonings, batches were prepared using twice the amount of a specific seasoning. Trials were done and assessed three times. Proportion differences were

  19. Purifying hydrocarbons

    Energy Technology Data Exchange (ETDEWEB)

    Demoulins, H D; Garner, F H

    1923-02-07

    Hydrocarbon distillates, including natural gases and vapors produced by cracking hydrocarbon oils, are desulfurized etc. by treating the vapor with an aqueous alkaline solution of an oxidizing agent. The hydrocarbons may be previously purified by sulfuric acid. In examples aqueous solutions of sodium or calcium hydrochlorite containing 1.5 to 5.0 grams per liter of available chlorine and sufficient alkali to give an excess of 0.1 percent in the spent reagent are preheated to the temperature of the vapor, and either sprayed or atomized into the vapors near the outlet of the dephlegmator or fractionating tower, or passed in countercurrent to the vapors through one or a series of scrubbers.

  20. Serodiagnosis of human neurocysticercosis using antigenic components of Taenia solium metacestodes derived from the unbound fraction from jacalin affinity chromatography

    Directory of Open Access Journals (Sweden)

    Gleyce Alves Machado

    2013-05-01

    Full Text Available The aim of the present study was to analyse Taenia solium metacestode antigens that were derived from the unbound fraction of jacalin affinity chromatography and subsequent tert-octylphenoxy poly (oxyethylene ethanol Triton X-114 (TX-114 partitioning in the diagnosis of human neurocysticercosis (NCC. Immunoassays were designed to detect T. solium-specific IgG antibodies by ELISA and immunoblot. Serum samples were collected from 132 individuals who were categorised as follows: 40 had NCC, 62 presented Taenia spp or other parasitic diseases and 30 were healthy individuals. The jacalin-unbound (J unbound fraction presented higher sensitivity and specificity rates than the jacalin-bound fraction and only this fraction was subjected to subsequent TX-114 partitioning, resulting in detergent (DJ unbound and aqueous (AJ unbound fractions. The ELISA sensitivity and specificity were 85% and 84.8% for J unbound , 92.5% and 93.5% for DJ unbound and 82.5% and 82.6% for AJ unbound . By immunoblot, the DJ unbound fraction showed 100% sensitivity and specificity and only serum samples from patients with NCC recognised the 50-70 kDa T. solium-specific components. We conclude that the DJ unbound fraction can serve as a useful tool for the differential immunodiagnosis of NCC by immunoblot.

  1. An Experiment with Air Purifiers in Delhi during Winter 2015-2016.

    Science.gov (United States)

    Vyas, Sangita; Srivastav, Nikhil; Spears, Dean

    2016-01-01

    Particulate pollution has important consequences for human health, and is an issue of global concern. Outdoor air pollution has become a cause for alarm in India in particular because recent data suggest that ambient pollution levels in Indian cities are some of the highest in the world. We study the number of particles between 0.5μm and 2.5μm indoors while using affordable air purifiers in the highly polluted city of Delhi. Though substantial reductions in indoor number concentrations are observed during air purifier use, indoor air quality while using an air purifier is frequently worse than in cities with moderate pollution, and often worse than levels observed even in polluted cities. When outdoor pollution levels are higher, on average, indoor pollution levels while using an air purifier are also higher. Moreover, the ratio of indoor air quality during air purifier use to two comparison measures of air quality without an air purifier are also positively correlated with outdoor pollution levels, suggesting that as ambient air quality worsens there are diminishing returns to improvements in indoor air quality during air purifier use. The findings of this study indicate that although the most affordable air purifiers currently available are associated with significant improvements in the indoor environment, they are not a replacement for public action in regions like Delhi. Although private solutions may serve as a stopgap, reducing ambient air pollution must be a public health and policy priority in any region where air pollution is as high as Delhi's during the winter.

  2. Steroidogenesis in amlodipine treated purified Leydig cells

    Energy Technology Data Exchange (ETDEWEB)

    Latif, Rabia, E-mail: rabialatif08@hotmail.com [Department of Physiology, Army Medical College, National University of Sciences and Technology, Islamabad (Pakistan); Lodhi, Ghulam Mustafa, E-mail: drmustafa786@gmail.com [Department of Physiology, Wah Medical College, Wah (Pakistan); Hameed, Waqas, E-mail: waqham@hotmail.com [Department of Physiology, Rehman Medical College, Peshawar (Pakistan); Aslam, Muhammad, E-mail: professormaslam@yahoo.com [Department of Physiology, Shifa College of Medicine, Islamabad (Pakistan)

    2012-01-01

    Drugs have been shown to adversely affect male fertility and recently anti-hypertensive drugs were added to the list. The anti-fertility effects of amlodipine, a calcium channel blocker, are well-illustrated in in vivo experiments but lack an in vitro proof. The present study was designed to experimentally elucidate the effects of amlodipine on Leydig cell steroidogenesis and intracellular calcium in vitro. Leydig cells of Sprague–Dawley rats were isolated and purified by Percoll. Cells were incubated for 3 h with/without amlodipine in the presence/absence of LH, dbcAMP, Pregnenolone and 25-Hydroxycholesterol. Cytosolic calcium was measured in purified Leydig cells by fluorometric technique. The results showed significantly reduced (P < 0.05) steroidogenesis and intracellular calcium in amlodipine exposed rats. The site of amlodipine induced steroidogenic inhibition seems to be prior to the formation of Pregnenolone at the level of StAR protein. -- Highlights: ► Inhibition of steroidogenesis in isolated and purified Leydig cells by amlodipine. ► Site of inhibition was before Pregnenolone formation, at the level of StAR protein. ► Inhibition of LH stimulated rise in cytosolic calcium by amlodipine.

  3. Development of a biogas purifier for rural areas in Japan

    Energy Technology Data Exchange (ETDEWEB)

    Kimura, Y.; Hinata, T. [Hokkaido Central Agricultural Experiment Station, Hokkaido (Japan); Yasui, S. [Zukosha Co. Ltd., Obihiro, Hokkaido (Japan); Noguchi, N. [Hokkaido Univ., Sapporo, Hokkaido (Japan); Tsukamoto, T. [IHI Shibaura. Co. Ltd., Obihiro, Hokkaido (Japan); Imai, T. [Green Plan Co. Ltd., Sapporo, Hokkaido (Japan); Kanai, M. [Air Water Co. Ltd, Sakai, Osaka (Japan); Matsuda, Z. [Hokuren Agricultural Research Center, Sapporo, Hokkaido (Japan)

    2010-07-01

    Although the biogas that is currently produced for dairy farms in Japan is a carbon-neutral energy, its use is restricted to farming areas only because there is no effective method of transporting unused biogas. There is a need for establishing practical methods for biogas removal from operating systems. In this study, a gas separation membrane was used in order to modify biogas to city gas 12A specifications, and to develop a biogas purifier equipped with a device to fill high pressure purified gas into cylinders to be taken outside the farming area. The objective was to expand the use of biogas produced from stand-alone gas plants. The amount of purified gas produced at a newly created refining-compression-filling (RCF) facility was approximately 97.0 Nm{sup 3}/day, for a raw material amount of about 216.0 Nm{sup 3}/day. The heat quantity of the purified gas was 38.9 MJ/Nm{sup 3}, which was within city gas 12A specifications. A total of 14.3 cylinders were filled each day with the manufactured purified gas. Test calculations along with a simulation exercise revealed that it would be possible to provide purified gas to approximately 6 per cent of common residences in a town in northern Japan. It was concluded that the newly created RCF facility allowed the modification of carbon-neutral biogas to conform to city gas 12A specifications, and allowed the transport of this gas out of the farming area.

  4. Method for purifying bidentate organophosphorus compounds

    International Nuclear Information System (INIS)

    Schulz, W.W.

    1977-01-01

    Bidentate organophosphorus compounds useful for extracting actinide elements from acidic nuclear waste solutions are purified of undesirable acidic impurities by contacting the compounds with ethylene glycol which preferentially extracts the impurities found in technical grade bidentate compounds

  5. Process for purifying zirconium sponge

    International Nuclear Information System (INIS)

    Abodishish, H.A.M.; Kimball, L.S.

    1992-01-01

    This patent describes a Kroll reduction process wherein a zirconium sponge contaminated with unreacted magnesium and by-product magnesium chloride is produced as a regulus, a process for purifying the zirconium sponge. It comprises: distilling magnesium and magnesium chloride from: a regulus containing a zirconium sponge and magnesium and magnesium chloride at a temperature above about 800 degrees C and at an absolute pressure less than about 10 mmHg in a distillation vessel to purify the zirconium sponge; condensing the magnesium and the magnesium chloride distilled from the zirconium sponge in a condenser; and then backfilling the vessel containing the zirconium sponge and the condenser containing the magnesium and the magnesium chloride with a gas; recirculating the gas between the vessel and the condenser to cool the zirconium sponge from above about 800 degrees C to below about 300 degrees C; and cooling the recirculating gas in the condenser containing the condensed magnesium and the condensed magnesium chloride as the gas cools the zirconium sponge to below about 300 degrees C

  6. Measurement of Ozone Emission and Particle Removal Rates from Portable Air Purifiers

    Science.gov (United States)

    Mang, Stephen A.; Walser, Maggie L.; Nizkorodov, Sergey A.; Laux, John M.

    2009-01-01

    Portable air purifiers are popular consumer items, especially in areas with poor air quality. Unfortunately, most users of these air purifiers have minimal understanding of the factors affecting their efficiency in typical indoor settings. Emission of the air pollutant ozone (O[subscript 3]) by certain air purifiers is of particular concern. In an…

  7. Purifying hydrocarbons in the gaseous stage

    Energy Technology Data Exchange (ETDEWEB)

    1937-02-01

    Gaseous tar oils are subjected, at temperatures of 320 to 380/sup 0/C, to the action of a mixture of activated carbon mixed with powdered metal which removes the sulfur contamination from the substance to be purified.

  8. Purified water quality study

    International Nuclear Information System (INIS)

    Spinka, H.; Jackowski, P.

    2000-01-01

    Argonne National Laboratory (HEP) is examining the use of purified water for the detection medium in cosmic ray sensors. These sensors are to be deployed in a remote location in Argentina. The purpose of this study is to provide information and preliminary analysis of available water treatment options and associated costs. This information, along with the technical requirements of the sensors, will allow the project team to determine the required water quality to meet the overall project goals

  9. Purifying oils

    Energy Technology Data Exchange (ETDEWEB)

    1930-04-15

    Gasoline, lamp oils, and lubricating or other mineral or shale oils are refined by contacting the vapor with a hot aqueous solution of salts of zinc, cadmium, or mercury, or mixtures thereof which may contain 0-5-3-0 percent of oxide or hydroxide in solution or suspension. Chlorides, bromides, iodides, sulfates, nitrates, and sulfonates of benzol, toluol, xylol, and petroleum are specified. Washing with a solution of sodium or potassium hydroxide or carbonate of calcium hydroxide may follow. The oil may first be purified by sulfuric acid or other known agent, or afterwards caustic alkali and sulfuric acid. The Specification as open to inspection under Sect. 91 (3) (a) describes also the use of salts of copper, iron, chromium, manganese, aluminum, nickel, or cobalt, with or without their oxides or hydroxides. This subject-matter does not appear in the Specification as accepted.

  10. Impossibility criterion for obtaining pure entangled states from mixed states by purifying protocols

    International Nuclear Information System (INIS)

    Chen Pingxing; Liang Linmei; Li Chengzu; Huang Mingqiu

    2002-01-01

    Purifying noisy entanglement is a protocol that can increase the entanglement of a mixed state (as a source) at the expense of the entanglement of others (such as an ancilla) by collective measurement. A protocol with which one can get a pure entangled state from a mixed state is defined as purifying mixed states. We address a basic question: can one get a pure entangled state from a mixed state? We give a necessary and sufficient condition of purifying a mixed state by fit local operations and classical communication and show that for a class of source states and ancilla states in arbitrary bipartite systems purifying mixed states is impossible by finite rounds of purifying protocols. For 2x2 systems, it is proved that arbitrary states cannot be purified by individual measurement. The possible application and meaning of the conclusion are discussed

  11. The taruca (Hippocamelus antisensis) and the red brocket deer (Mazama americana) as intermediate hosts of Taenia hydatigena in Peru, morphological and molecular evidence.

    Science.gov (United States)

    Gomez-Puerta, Luis A; Pacheco, Joel; Gonzales-Viera, Omar; Lopez-Urbina, Maria T; Gonzalez, Armando E

    2015-09-15

    In the present report metacestodes were collected from the mesentery of a taruca (Hippocamelus antisensis) and from the omentum of a red brocket deer (Mazama americana) in Peru. Various metacestodes parameters, including rostellar hook characteristics, were measured. Molecular analysis was performed to amplify the mitochondrial cytochrome c oxidase subunit 1 gene from metacestode isolates. Metacestodes were identified as T. hydatigena by morphology and molecular methods. This constitutes the first molecular detection of T. hydatigena metacestodes in the taruca and the red brocket deer and demonstrates that these animal species are natural intermediate hosts for this parasite. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Home drinking-water purifiers

    International Nuclear Information System (INIS)

    Pizzichini, Massimo; Pozio, Alfonso; Russo, Claudio

    2005-01-01

    To salve the widespread problem of contaminated drinking water, home purifiers are now sold in Italy as well as other countries. This article describes how these devices work, how safe they are to use and how safe the water they produce, in the broad context of regulations on drinking water and mineral water. A new device being developed by ENEA to treat municipal water and ground water could provide greater chemical and bacteriological safety. However, the appearance of these new systems makes it necessary to update existing regulations [it

  13. Robust sparse image reconstruction of radio interferometric observations with PURIFY

    Science.gov (United States)

    Pratley, Luke; McEwen, Jason D.; d'Avezac, Mayeul; Carrillo, Rafael E.; Onose, Alexandru; Wiaux, Yves

    2018-01-01

    Next-generation radio interferometers, such as the Square Kilometre Array, will revolutionize our understanding of the Universe through their unprecedented sensitivity and resolution. However, to realize these goals significant challenges in image and data processing need to be overcome. The standard methods in radio interferometry for reconstructing images, such as CLEAN, have served the community well over the last few decades and have survived largely because they are pragmatic. However, they produce reconstructed interferometric images that are limited in quality and scalability for big data. In this work, we apply and evaluate alternative interferometric reconstruction methods that make use of state-of-the-art sparse image reconstruction algorithms motivated by compressive sensing, which have been implemented in the PURIFY software package. In particular, we implement and apply the proximal alternating direction method of multipliers algorithm presented in a recent article. First, we assess the impact of the interpolation kernel used to perform gridding and degridding on sparse image reconstruction. We find that the Kaiser-Bessel interpolation kernel performs as well as prolate spheroidal wave functions while providing a computational saving and an analytic form. Secondly, we apply PURIFY to real interferometric observations from the Very Large Array and the Australia Telescope Compact Array and find that images recovered by PURIFY are of higher quality than those recovered by CLEAN. Thirdly, we discuss how PURIFY reconstructions exhibit additional advantages over those recovered by CLEAN. The latest version of PURIFY, with developments presented in this work, is made publicly available.

  14. Process for purifying graphite

    International Nuclear Information System (INIS)

    Clausius, R.A.

    1985-01-01

    A process for purifying graphite comprising: comminuting graphite containing mineral matter to liberate at least a portion of the graphite particles from the mineral matter; mixing the comminuted graphite particles containing mineral matter with water and hydrocarbon oil to form a fluid slurry; separating a water phase containing mineral matter and a hydrocarbon oil phase containing grahite particles; and separating the graphite particles from the hydrocarbon oil to obtain graphite particles reduced in mineral matter. Depending upon the purity of the graphite desired, steps of the process can be repeated one or more times to provide a progressively purer graphite

  15. Home Air Purifiers Eradicate Harmful Pathogens

    Science.gov (United States)

    2014-01-01

    Marshall Space Flight Center funded the University of Madison-Wisconsin to develop ethylene scrubbers to keep produce fresh in space. Akida Holdings of Jacksonville, Florida, licensed the technology and developed Airocide, an air purifier that can kill airborne pathogens. Previously designed for industrial spaces, there is now a specially designed unit for home use.

  16. Guanine nucleotide regulatory protein co-purifies with the D2-dopamine receptor

    International Nuclear Information System (INIS)

    Senogles, S.E.; Caron, M.G.

    1986-01-01

    The D 2 -dopamine receptor from bovine anterior pituitary was purified ∼1000 fold by affinity chromatography on CMOS-Sepharose. Reconstitution of the affinity-purified receptor into phospholipid vesicles revealed the presence of high and low affinity agonist sites as detected by N-n-propylnorapomorphine (NPA) competition experiments with 3 H-spiperone. High affinity agonist binding could be converted to the low affinity form by guanine nucleotides, indicating the presence of an endogenous guanine nucleotide binding protein (N protein) in the affinity-purified D 2 receptor preparations. Furthermore, this preparation contained an agonist-sensitive GTPase activity which was stimulated 2-3 fold over basal by 10 μM NPA. 35 S-GTPγS binding to these preparations revealed a stoichiometry of 0.4-0.7 mole N protein/mole receptor, suggesting the N protein may be specifically coupled with the purified D 2 -dopamine receptor and not present as a contaminant. Pertussis toxin treatment of the affinity purified receptor preparations prevented high affinity agonist binding, as well as agonist stimulation of the GTPase activity, presumably by inactivating the associated N protein. Pertussis toxin lead to the ADP-ribosylation of a protein of 39-40K on SDS-PAGE. These findings indicate that an endogenous N protein, N/sub i/ or N/sub o/, co-purifies with the D 2 -dopamine receptor which may reflect a precoupling of this receptor with an N protein within the membranes

  17. Mine water purify from radium

    International Nuclear Information System (INIS)

    Lebecka, J.

    1996-01-01

    The article describes purification of radium containing water in coal mines. Author concludes that water purification is relatively simple and effective way to decrease environmental pollution caused by coal mining. The amount of radium disposed with type A radium water has been significantly decreased. The results of investigations show that it will be soon possible to purify also type B radium water. Article compares the amounts of radium disposed by coal mines in 1990, 1995 and forecast for 2000

  18. Simultaneous purifying of Hg0, SO2, and NOx from flue gas by Fe3+/H2O2: the performance and purifying mechanism.

    Science.gov (United States)

    Xing, Yi; Li, Liuliu; Lu, Pei; Cui, Jiansheng; Li, Qianli; Yan, Bojun; Jiang, Bo; Wang, Mengsi

    2018-03-01

    Hg 0 , SO 2 , and NOx result in heavily global environmental pollution and serious health hazards. Up to now, how to efficiently remove mercury with SO 2 and NOx from flue gas is still a tough task. In this study, series of high oxidizing Fenton systems were employed to purify the pollutants. The experimental results showed that Fe 3+ /H 2 O 2 was more suitable to purify Hg 0 than Fe 2+ /H 2 O 2 and Cu 2+ /H 2 O 2. The optimal condition includes Fe 3+ concentration of 0.008 mol/L, Hg 0 inlet concentration of 40 μg/m 3 , solution temperature of 50 °C, pH of 3, H 2 O 2 concentration of 0.7 mol/L, and O 2 percentage of 6%. When SO 2 and NOx were taken into account under the optimal condition, Hg 0 removal efficiency could be enhanced to 91.11% while the removal efficiency of both NOx and SO 2 was slightly declined, which was consistent to the analysis of purifying mechanism. The removal efficiency of Hg 0 was stimulated by accelerating the conversion of Fe 2+ to Fe 3+ , which resulted from the existence of SO 2 and NOx. The results of this study suggested that simultaneously purifying Hg 0 , SO 2 , and NOx from flue gas is feasible.

  19. Studies on a novel peptide isolated and purified from rat insulinoma tissue

    Energy Technology Data Exchange (ETDEWEB)

    Al-Akhras, G N

    1987-01-01

    Rat insulinoma peptide (RIP) which appears to be either a fragment of, or an altered rat C-peptide was isolated and purified by dialysis. The purity of this peptide was investigated using polyacrylamide gel electrophoresis with sodium dodecyl sulfate, isoelectric focusing, and high performance liquid chromatography. RIP may contain two peptides similar to each other but differing in their isoelectric points. The molecular weight of RIP was found to be 1982 daltons by fast atoms bombardment mass spectrometry giving a chain length of approximately 22 amino acid residues. From information obtained using radioimmunoassay employing antiserum R901, RIP appears to share a common C-terminus with rat C-peptide. A radioimmunoassay for RIP was developed using the purified RIP as immunogen and for standards and tracers. An indirect enzyme linked immunosorbent assay (ELISA) for rat insulinoma peptide was developed using purified RIP for immunogen and semi-purified RIP as a standard.

  20. Cooling performance of R510A in domestic water purifiers

    International Nuclear Information System (INIS)

    Park, Ki Jung; Lee, Yo Han; Jung, Dong Soo

    2010-01-01

    Cooling performance of R510A is examined both numerically and experimentally in an effort to replace HFC134a in the refrigeration system of domestic water purifiers. Although the use of HFC134a is currently dominant, it is being phased out in Europe and most developed countries due to its high potential contribution to global warming. To solve this problem, cycle simulation and experimental measurements are conducted with a new refrigerant mixture of 88%RE170/12%R600a using actual domestic water purifiers. This mixture has been recently numbered and listed as R510A by ASHRAE. Test results show that, due to the small internal volume of the refrigeration system of the domestic water purifiers, system performance with R510A is greatly influenced by the amount of charge. With the optimum charge amount of 20 to 21 g, approximately 50% that of HFC134a, the energy consumption of R510A is 22.3% lower than that of HFC134a. The compressor discharge temperature of R510A is 3.7 .deg. C lower than that of HFC134a at the optimum charge. Overall, R510A, a new, long term, and environmentally safe refrigerant, is a good alternative for HFC134a. Furthermore, it requires only minor changes in the refrigeration system of the domestic water purifiers

  1. Current status of taeniasis in Thailand.

    Science.gov (United States)

    Anantaphruti, Malinee Thairungroj

    2013-02-01

    Taeniasis is prevalent in all regions of Thailand, except the South. Infections were more frequently found in males than females of any age from 7-83 years. Taenia saginata is the most common species throughout the country. Taenia asiatica was reported only in the province of Kanchanaburi in the Central region. Co-infections, with Taenia solium and T. asiatica or T. solium and T. saginata, were found. Hybridization between T. asiatica and T. saginata is evidence that co-infection is never found between these 2 species. Finding more than 1 worm in a single patient was not entirely rare. Genetic variation was found without correlation to its geographic distribution in T. saginata, whereas no variation was found in T. asiatica.

  2. Characterization and treatment of cyanide in MGP purifier wastes

    Energy Technology Data Exchange (ETDEWEB)

    Theis, T.L. [Clarkson University, Potsdam, NY (United States). Dept. of Civil and Environmental Engineering

    1995-12-31

    Purifier wastes were generated from the clean-up gaseous impurities, principally hydrogen sulfide and hydrogen cyanide, contained in raw gas from MGP operations through retention by iron oxide solids. These materials were generated at a rate of about 10-20 kg/1000 m{sup 3} of gas produced, and although regeneration was sometimes practised, eventual disposal as fill material, usually on site, was eventually necessary. The remediation of MGP sites generally requires that the disposition of these waste solids be addressed. The effective treatment of purifier wastes presents special problems due to the acid-base properties of the material, its elevated sulfur content, and the significant quantities of carbon both added as wood shavings and present as compounds generated as a result of gas manufacture. In broad terms, treatment approaches can be divided into two classes, those aimed at destroying the cyanide and objectionable carbon compounds and otherwise disposing of the residual, and those which attempt to isolate the waste from its surroundings. The latter approach attempts to take advantage of the natural insolubility of most of the constituents of concern found in purifier wastes, while destructive technologies limit potential liability. 9 refs.

  3. Full scale demonstration of air-purifying pavement

    NARCIS (Netherlands)

    Ballari, M.; Brouwers, H.J.H.

    2013-01-01

    Experiments concerning a full-scale demonstration of air purifying pavement in Hengelo, The Netherlands, are reported. The full width of the street was provided with concrete pavement containing TiO2 over a length of 150 m ("DeNOx street"). Another part of the street, about 100 m, was paved with

  4. [Studies on the process of Herba Clinopodii saponins purified with macroporous adsorption resin].

    Science.gov (United States)

    Zhang, Yi; Yan, Dan; Han, Yumei

    2005-10-01

    To study the technological parameters of the purification process of saponins with macroporous adsorption resin. The adsorptive characteristics and elutive parameters of the process were studied by taking the elutive and purified ratio of saponins as markers. 11.4 ml of the extraction of Herba Clinopodii (crude drugs 0.2 g/ml) was purified with a column of macroporous adsorption resin (phi15 mm x H90 mm, dry weight 2.5 g) and washed with 3BV of distilled water, then eluted with 3BV of 30% ethanol and 3BV of 70% ethanol. Most of saponins were collected in the 70% ethanol. With macroporous adsorption resin adsorbing and purifying,the elutive ratio of saponins is 86.8% and the purity reaches 153.2%. So this process of applying macroporous adsorption resin to adsorb and purify Saponins is feasible.

  5. 76 FR 29191 - Purified Carboxymethylcellulose From Finland and the Netherlands: Continuation of Antidumping...

    Science.gov (United States)

    2011-05-20

    ... Carboxymethylcellulose From Finland and the Netherlands: Continuation of Antidumping Duty Orders AGENCY: Import... antidumping duty orders on purified carboxymethylcellulose from Finland and the Netherlands would likely lead...) from Finland and the Netherlands. See Notice of Antidumping Duty Orders: Purified...

  6. 78 FR 9884 - Purified Carboxymethylcellulose From the Netherlands: Final Results of Antidumping Duty...

    Science.gov (United States)

    2013-02-12

    ... Carboxymethylcellulose From the Netherlands: Final Results of Antidumping Duty Administrative Review and Final No... carboxymethylcellulose (purified CMC) from the Netherlands.\\1\\ This review covers two respondents, Akzo Nobel Functional... Review'' section of this notice. \\1\\ See Purified Carboxymethylcellulose From the Netherlands...

  7. Proteomic analysis of purified coronavirus infectious bronchitis virus particles

    Directory of Open Access Journals (Sweden)

    Shu Dingming

    2010-06-01

    Full Text Available Abstract Background Infectious bronchitis virus (IBV is the coronavirus of domestic chickens causing major economic losses to the poultry industry. Because of the complexity of the IBV life cycle and the small number of viral structural proteins, important virus-host relationships likely remain to be discovered. Toward this goal, we performed two-dimensional gel electrophoresis fractionation coupled to mass spectrometry identification approaches to perform a comprehensive proteomic analysis of purified IBV particles. Results Apart from the virus-encoded structural proteins, we detected 60 host proteins in the purified virions which can be grouped into several functional categories including intracellular trafficking proteins (20%, molecular chaperone (18%, macromolcular biosynthesis proteins (17%, cytoskeletal proteins (15%, signal transport proteins (15%, protein degradation (8%, chromosome associated proteins (2%, ribosomal proteins (2%, and other function proteins (3%. Interestingly, 21 of the total host proteins have not been reported to be present in virions of other virus families, such as major vault protein, TENP protein, ovalbumin, and scavenger receptor protein. Following identification of the host proteins by proteomic methods, the presence of 4 proteins in the purified IBV preparation was verified by western blotting and immunogold labeling detection. Conclusions The results present the first standard proteomic profile of IBV and may facilitate the understanding of the pathogenic mechanisms.

  8. 21 CFR 880.6500 - Medical ultraviolet air purifier.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Medical ultraviolet air purifier. 880.6500 Section 880.6500 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... to ultraviolet radiation. (b) Classification. Class II (performance standards). ...

  9. Ultrasonic-resonator-combined apparatus for purifying nuclear aerosol particles

    Energy Technology Data Exchange (ETDEWEB)

    Hou, Suxia; Zhang, Quanhu; Li, Sufen; Chen, Chen; Su, Xianghua [Xi' an Hi-Tech Institute, Xi' an (China)

    2017-12-15

    The radiation hazards of radionuclides in the air arising from the storage room of nuclear devices to the operators cannot be ignored. A new ultrasonic-resonator-combined method for purifying nuclear aerosol particles is introduced. To remove particles with diameters smaller than 0.3 μm, an ultrasonic chamber is induced to agglomerate these submicron particles. An apparatus which is used to purify the nuclear aerosol particles is described in the article. The apparatus consists of four main parts: two filtering systems, an ultrasonic chamber and a high-pressure electrostatic precipitator system. Finally, experimental results demonstrated the effectiveness of the implementation of the ultrasonic resonators. The feasibility of the method is proven by its application to the data analysis of the experiments.

  10. Effect of streamer plasma air purifier on sbs symptoms and performance of office work

    DEFF Research Database (Denmark)

    Zhang, X.J.; Fang, Lei; Wargocki, Pawel

    2011-01-01

    Subjective experiments were conducted to evaluate the effect of a streamer plasma air purifier on perceived air quality, SBS symptoms and performance of office work during 5-hour exposure of 32 recruited subjects in field laboratory in which real materials were used to establishing a realistic...... level of air pollution. Intensity of SBS symptoms were indicated using visual-analogue scales. Subjects’ performance was evaluated with several computer tasks. The results show that operation of the air purifiers improved perceived air quality and reduced the odor intensity of indoor air. Eye dryness...... symptom was found significantly improved when the air purifiers were used but no other SBS symptoms or performance of office work were improved when the air purifiers were in operation compared to the condition when they were off....

  11. Comparison of single-step and two-step purified coagulants from Moringa oleifera seed for turbidity and DOC removal.

    Science.gov (United States)

    Sánchez-Martín, J; Ghebremichael, K; Beltrán-Heredia, J

    2010-08-01

    The coagulant proteins from Moringa oleifera purified with single-step and two-step ion-exchange processes were used for the coagulation of surface water from Meuse river in The Netherlands. The performances of the two purified coagulants and the crude extract were assessed in terms of turbidity and DOC removal. The results indicated that the optimum dosage of the single-step purified coagulant was more than two times higher compared to the two-step purified coagulant in terms of turbidity removal. And the residual DOC in the two-step purified coagulant was lower than in single-step purified coagulant or crude extract. (c) 2010 Elsevier Ltd. All rights reserved.

  12. Effect of partially purified angiotensin converting enzyme inhibitory ...

    African Journals Online (AJOL)

    This study evaluated the effect of partially-purified angiotensin converting enzyme (ACE) inhibitory proteins obtained from the leaves of Moringa oleifera on blood glucose, serum ACE activity and lipid profile of alloxaninduced diabetic rats. Twenty-five apparently healthy male albino rats were divided into five groups of five ...

  13. HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans.

    Science.gov (United States)

    Flores, María D; Gonzalez, Luis M; Hurtado, Carolina; Motta, Yamileth Monje; Domínguez-Hidalgo, Cristina; Merino, Francisco Jesús; Perteguer, María J; Gárate, Teresa

    2018-02-27

    Taenia solium, T. asiatica and T. saginata tapeworms cause human taeniasis and are the origin of porcine and bovine cysticercosis. Furthermore, T. solium eggs can cause human cysticercosis, with neurocysticercosis being the most serious form of the disease. These helminth infections are neglected tropical diseases and are endemic in several countries in the Americas, Asia and Africa. As a result of globalization, migration in particular, the infections have been extending to non-endemic territories. Species-specific diagnosis of taeniasis is subject to drawbacks that could be resolved using molecular approaches. In the present study, conventional and real-time amplification protocols (cPCR and qPCR) based on the T. saginata HDP2 sequence were applied in the differential diagnosis of taeniasis (T. saginata, T. solium) in both fecal samples and proglottids expelled by patients. The HDP2 homolog in T. solium was cloned and characterized. Semi-nested cPCR and qPCR (Sn-HDP2 cPCR and Sn-HDP2 qPCR) amplified T. saginata and T. solium DNA, with an analytical sensitivity of 40 and 400 fg, respectively, and identically in both protocols. Eighteen taeniasis patients were diagnosed directly with T. saginata or T. solium, either from proglottids or fecal samples with/without eggs (detected using microscopy), based on the optimized Sn-HDP2 qPCR. After cloning, the T. solium HDP2 homolog sequence was confirmed to be a ribosomal sequence. The HDP2 fragment corresponded to a non-transcribed sequence/external transcribed repeat (NTS/ETS) of ribosomal DNA. Compared with the T. saginata HDP2 homolog, the T solium HDP2 sequence lacked the first 900 nt at the 5' end and showed nucleotide substitutions and small deletions. Sn-HDP2 cPCR and Sn-HDP2 qPCR were set up for the diagnosis of human taeniasis, using proglottids and fecal samples from affected patients. The new Sn-HDP2 qPCR protocol was the best option, as it directly differentiated T. saginata from T. solium. The diagnosis of

  14. Can a photocatalytic air purifier be used to improve the perceived air quality indoors?

    DEFF Research Database (Denmark)

    Kolarik, Jakub; Wargocki, Pawel

    2010-01-01

    The effect of a photocatalytic air purifier on perceived air quality(PAQ) was examined in rooms polluted by typical sources of indoor pollution.The rooms were ventilated at three different outdoor air supply rates. The air quality was assessed by a sensory panel when the purifier was in operation...... as well as when it was off. Operation of the purifier significantly improved PAQ in the rooms polluted by building materials (used carpet, old linoleum, and old chip-board), and a used ventilation filter as well as a mixture of building materials, used ventilation filter and cathode-ray tube computer...... monitors. The effect cor-responded to approximately doubling the outdoor air supply rate. Operation of the purifier significantly worsened the PAQ in rooms with human bioeffluents, probably due to incomplete oxidation of alcohols which are one of the main pollutants emitted by humans. Present results show...

  15. Effect of Amphiphilic Alkyl Chain Length Upon Purified LATEX Stability

    International Nuclear Information System (INIS)

    Amira Amir Hassan; Amir Hashim Mohd Yatim

    2015-01-01

    Rubber particles in purified latex (PL) are stabilized by a film of protein and fatty acid soap (surfactant). Saturated straight-chain fatty acid soaps can assist an enhancement of latex stability. However, whether the alkyl chain length plays an important role in increasing the stability is still an issue. The aim of this study is to investigate the effect of alkyl chain length of anionic surfactant on the stability of purified latex. The fatty acid soap of decanoate (9), laurate (11), sodium dodecyl sulphate (SDS) (12) and palmitate (15) were used. The numbers in parentheses indicating the number of carbon present in alkyl chain of the soap. The results showed that the impact of alkyl chain length on the stability of latex is in the order of laurate > decanoate > SDS > palmitate > purified latex accordingly. The alkyl chain length does giving a significant effect on latex stability after longer stirring time. The particle size of latex with the presence of surfactant is greater compare to a single particle itself due to extension of particles diameter. Thus suitable interaction of the nonpolar tail of surfactant with the hydrophobic regions of latex surface played a major role in maintaining a stable latex system. (author)

  16. Extraction and characterization of highly purified collagen from bovine pericardium for potential bioengineering applications

    International Nuclear Information System (INIS)

    Santos, Maria Helena; Silva, Rafael M.; Dumont, Vitor C.; Neves, Juliana S.; Mansur, Herman S.; Heneine, Luiz Guilherme D.

    2013-01-01

    Bovine pericardium is widely used as a raw material in bioengineering as a source of collagen, a fundamental structural molecule. The physical, chemical, and biocompatibility characteristics of these natural fibers enable their broad use in several areas of the health sciences. For these applications, it is important to obtain collagen of the highest possible purity. The lack of a method to produce these pure biocompatible materials using simple and economically feasible techniques presents a major challenge to their production on an industrial scale. This study aimed to extract, purify, and characterize the type I collagen protein originating from bovine pericardium, considered to be an abundant tissue resource. The pericardium tissue was collected from male animals at slaughter age. Pieces of bovine pericardium were enzymatically digested, followed by a novel protocol developed for protein purification using ion-exchange chromatography. The material was extensively characterized by electrophoresis, scanning electron microscopy, energy dispersive X-ray spectroscopy, and infrared spectroscopy. The results showed a purified material with morphological properties and chemical functionalities compatible with type I collagen and similar to a highly purified commercial collagen. Thus, an innovative and relatively simple processing method was developed to extract and purify type I collagen from bovine tissue with potential applications as a biomaterial for regenerative tissue engineering. - Highlights: ► Type I collagen was obtained from bovine pericardium, an abundant tissue resource. ► A simple and feasible processing technique was developed to purify bovine collagen. ► The appropriate process may be performed on industrial scale. ► The pure collagen presented appropriate morphological and molecular characteristics. ► The purify collagen has shown potential use as a biomaterial in tissue engineering.

  17. Studying the fate of non-volatile organic compounds in a commercial plasma air purifier

    Energy Technology Data Exchange (ETDEWEB)

    Schmid, Stefan [ETH Zürich, Department of Chemistry and Applied Biosciences, CH-8093 Zürich (Switzerland); Seiler, Cornelia; Gerecke, Andreas C. [Swiss Federal Laboratories for Material Science and Technology (EMPA), CH-8600 Dübendorf (Switzerland); Hächler, Herbert [University of Zürich, Institute for Food Safety and Hygiene, National Centre for Enteropathogenic Bacteria and Listeria (NENT), CH-8057 Zürich (Switzerland); Hilbi, Hubert [Ludwig-Maximilians-Universität München Max von Pettenkofer-Institut, D-80336 München (Germany); Frey, Joachim [University of Bern, Institute for Veterinary Bacteriology, CH-3001 Bern (Switzerland); Weidmann, Simon; Meier, Lukas; Berchtold, Christian [ETH Zürich, Department of Chemistry and Applied Biosciences, CH-8093 Zürich (Switzerland); Zenobi, Renato, E-mail: zenobi@org.chem.ethz.ch [ETH Zürich, Department of Chemistry and Applied Biosciences, CH-8093 Zürich (Switzerland)

    2013-07-15

    Highlights: • Degradation of environmental toxins, a protein, and bioparticles were studied. • A commercial air purifier based on a cold plasma was used. • Passage through the device reduced the concentration of the compounds/particles. • Deposition inside the plasma air purifier was the main removal process. -- Abstract: Degradation of non-volatile organic compounds–environmental toxins (methyltriclosane and phenanthrene), bovine serum albumin, as well as bioparticles (Legionella pneumophila, Bacillus subtilis, and Bacillus anthracis)–in a commercially available plasma air purifier based on a cold plasma was studied in detail, focusing on its efficiency and on the resulting degradation products. This system is capable of handling air flow velocities of up to 3.0 m s{sup −1} (3200 L min{sup −1}), much higher than other plasma-based reactors described in the literature, which generally are limited to air flow rates below 10 L min{sup −1}. Mass balance studies consistently indicated a reduction in concentration of the compounds/particles after passage through the plasma air purifier, 31% for phenanthrene, 17% for methyltriclosane, and 80% for bovine serum albumin. L. pneumophila did not survive passage through the plasma air purifier, and cell counts of aerosolized spores of B. subtilis and B. anthracis were reduced by 26- and 15-fold, depending on whether it was run at 10 Hz or 50 Hz, respectively. However rather than chemical degradation, deposition on the inner surfaces of the plasma air purifier occured. Our interpretation is that putative “degradation” efficiencies were largely due to electrostatic precipitation rather than to decomposition into smaller molecules.

  18. Biological properties of purified recombinant HCV particles with an epitope-tagged envelope

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Hitoshi; Akazawa, Daisuke [Department of Virology II, National Institute of Infectious Diseases, Tokyo (Japan); Toray Industries, Inc., Kanagawa (Japan); Kato, Takanobu; Date, Tomoko [Department of Virology II, National Institute of Infectious Diseases, Tokyo (Japan); Shirakura, Masayuki [Department of Virology II, National Institute of Infectious Diseases, Tokyo (Japan); Toray Industries, Inc., Kanagawa (Japan); Nakamura, Noriko; Mochizuki, Hidenori [Toray Industries, Inc., Kanagawa (Japan); Tanaka-Kaneko, Keiko; Sata, Tetsutaro [Department of Pathology, National Institute of Infectious Diseases, Tokyo (Japan); Tanaka, Yasuhito [Department of Clinical Molecular Informative Medicine, Nagoya City University Graduate School of Medicine, Nagoya (Japan); Mizokami, Masashi [Research Center for Hepatitis and Immunology, Kohnodai Hospital, International Medical Center of Japan, Chiba (Japan); Suzuki, Tetsuro [Department of Virology II, National Institute of Infectious Diseases, Tokyo (Japan); Wakita, Takaji, E-mail: wakita@nih.go.jp [Department of Virology II, National Institute of Infectious Diseases, Tokyo (Japan)

    2010-05-14

    To establish a simple system for purification of recombinant infectious hepatitis C virus (HCV) particles, we designed a chimeric J6/JFH-1 virus with a FLAG (FL)-epitope-tagged sequence at the N-terminal region of the E2 hypervariable region-1 (HVR1) gene (J6/JFH-1/1FL). We found that introduction of an adaptive mutation at the potential N-glycosylation site (E2N151K) leads to efficient production of the chimeric virus. This finding suggests the involvement of glycosylation at Asn within the envelope protein(s) in HCV morphogenesis. To further analyze the biological properties of the purified recombinant HCV particles, we developed a strategy for large-scale production and purification of recombinant J6/JFH-1/1FL/E2N151K. Infectious particles were purified from the culture medium of J6/JFH-1/1FL/E2N151K-infected Huh-7 cells using anti-FLAG affinity chromatography in combination with ultrafiltration. Electron microscopy of the purified particles using negative staining showed spherical particle structures with a diameter of 40-60 nm and spike-like projections. Purified HCV particle-immunization induced both an anti-E2 and an anti-FLAG antibody response in immunized mice. This strategy may contribute to future detailed analysis of HCV particle structure and to HCV vaccine development.

  19. Biological properties of purified recombinant HCV particles with an epitope-tagged envelope

    International Nuclear Information System (INIS)

    Takahashi, Hitoshi; Akazawa, Daisuke; Kato, Takanobu; Date, Tomoko; Shirakura, Masayuki; Nakamura, Noriko; Mochizuki, Hidenori; Tanaka-Kaneko, Keiko; Sata, Tetsutaro; Tanaka, Yasuhito; Mizokami, Masashi; Suzuki, Tetsuro; Wakita, Takaji

    2010-01-01

    To establish a simple system for purification of recombinant infectious hepatitis C virus (HCV) particles, we designed a chimeric J6/JFH-1 virus with a FLAG (FL)-epitope-tagged sequence at the N-terminal region of the E2 hypervariable region-1 (HVR1) gene (J6/JFH-1/1FL). We found that introduction of an adaptive mutation at the potential N-glycosylation site (E2N151K) leads to efficient production of the chimeric virus. This finding suggests the involvement of glycosylation at Asn within the envelope protein(s) in HCV morphogenesis. To further analyze the biological properties of the purified recombinant HCV particles, we developed a strategy for large-scale production and purification of recombinant J6/JFH-1/1FL/E2N151K. Infectious particles were purified from the culture medium of J6/JFH-1/1FL/E2N151K-infected Huh-7 cells using anti-FLAG affinity chromatography in combination with ultrafiltration. Electron microscopy of the purified particles using negative staining showed spherical particle structures with a diameter of 40-60 nm and spike-like projections. Purified HCV particle-immunization induced both an anti-E2 and an anti-FLAG antibody response in immunized mice. This strategy may contribute to future detailed analysis of HCV particle structure and to HCV vaccine development.

  20. Investigating the characteristic strength of flocs formed from crude and purified Hibiscus extracts in water treatment.

    Science.gov (United States)

    Jones, Alfred Ndahi; Bridgeman, John

    2016-10-15

    The growth, breakage and re-growth of flocs formed using crude and purified seed extracts of Okra (OK), Sabdariffa (SB) and Kenaf (KE) as coagulants and coagulant aids was assessed. The results showed floc size increased from 300 μm when aluminium sulphate (AS) was used as a coagulant to between 696 μm and 722 μm with the addition of 50 mg/l of OK, KE and SB crude samples as coagulant aids. Similarly, an increase in floc size was observed when each of the purified proteins was used as coagulant aid at doses of between 0.123 and 0.74 mg/l. The largest floc sizes of 741 μm, 460 μm and 571 μm were obtained with a 0.123 mg/l dose of purified Okra protein (POP), purified Sabdariffa (PSP) and purified Kenaf (PKP) respectively. Further coagulant aid addition from 0.123 to 0.74 mg/l resulted in a decrease in floc size and strength in POP and PSP. However, an increase in floc strength and reduced d50 size was observed in PKP at a dose of 0.74 mg/l. Flocs produced when using purified and crude extract samples as coagulant aids exhibited high recovery factors and strength. However, flocs exhibited greater recovery post-breakage when the extracts were used as a primary coagulant. It was observed that the combination of purified proteins and AS improved floc size, strength and recovery factors. Therefore, the applications of Hibiscus seeds in either crude or purified form increases floc growth, strength, recoverability and can also reduce the cost associated with the import of AS in developing countries. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

  1. Genetic diversity of Taenia asiatica from Thailand and other geographical locations as revealed by cytochrome c oxidase subunit 1 sequences.

    Science.gov (United States)

    Anantaphruti, Malinee Thairungroj; Thaenkham, Urusa; Watthanakulpanich, Dorn; Phuphisut, Orawan; Maipanich, Wanna; Yoonuan, Tippayarat; Nuamtanong, Supaporn; Pubampen, Somjit; Sanguankiat, Surapol

    2013-02-01

    Twelve 924 bp cytochrome c oxidase subunit 1 (cox1) mitochondrial DNA sequences from Taenia asiatica isolates from Thailand were aligned and compared with multiple sequence isolates from Thailand and 6 other countries from the GenBank database. The genetic divergence of T. asiatica was also compared with Taenia saginata database sequences from 6 different countries in Asia, including Thailand, and 3 countries from other continents. The results showed that there were minor genetic variations within T. asiatica species, while high intraspecies variation was found in T. saginata. There were only 2 haplotypes and 1 polymorphic site found in T. asiatica, but 8 haplotypes and 9 polymorphic sites in T. saginata. Haplotype diversity was very low, 0.067, in T. asiatica and high, 0.700, in T. saginata. The very low genetic diversity suggested that T. asiatica may be at a risk due to the loss of potential adaptive alleles, resulting in reduced viability and decreased responses to environmental changes, which may endanger the species.

  2. Mechanical performance of HMA-2 modified with purified and unpurified carbon nanotubes and nanofibers

    Directory of Open Access Journals (Sweden)

    Mario Rodrigo Rubio

    2017-05-01

    Full Text Available The present study evaluates the mechanical performance of a Hot Mix Asphalt – Type II (HMA-2 modified with carbon nanotubes and carbon nanofibers (CNTF. CNTF were made by means the Catalytic Vapor Deposition (CVD technique at 700° C using a Nickel, Copper and Aluminum (NiCuAl catalyst with a Cu/Ni molar relation of 0,33. In order to properly assess HMA-2 performance, three different mixtures were analyzed: 1 HMA-2 modified with purified CNTF; 2 HMA-2 modified with non-purified CNTF and, 3 a Conventional HMA-2 (control. Samples manufactured in accordance with the Marshall Mix Design were tested in the laboratory to study rutting, resilient modulus (Mr and fatigue. In addition to the aforementioned dynamic characterization, the effect of CNTF purification on the asphalt mixture’s mechanical properties was analyzed. In short, a comparative study was designed to determine whether or not CNTF should be purified before introduction into the HMA-2. This investigation responds to the growing demand for economical materials capable of withstanding traffic loads while simultaneously enhancing pavement durability and mechanical properties. Although purified CNTF increased HMA-2 stiffness and elastic modulus, non-purified CNTF increased the asphalt mixture’s elastic modulus without considerable increases in stiffness. Thus, the latter modification is deemed to help address fatiguerelated issues and improve the long-term durability of flexible pavements.

  3. Extraction and characterization of highly purified collagen from bovine pericardium for potential bioengineering applications

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Maria Helena, E-mail: mariahelena.santos@gmail.com [Department of Dentistry, Federal University of Vales do Jequitinhonha e Mucuri-UFVJM, Diamantina/MG 39100-000 (Brazil); Center for Assessment and Development of Biomaterials-BioMat, Federal University of Vales do Jequitinhonha e Mucuri-UFVJM, Diamantina/MG 39100-000 (Brazil); Silva, Rafael M.; Dumont, Vitor C. [Department of Dentistry, Federal University of Vales do Jequitinhonha e Mucuri-UFVJM, Diamantina/MG 39100-000 (Brazil); Center for Assessment and Development of Biomaterials-BioMat, Federal University of Vales do Jequitinhonha e Mucuri-UFVJM, Diamantina/MG 39100-000 (Brazil); Neves, Juliana S. [Center for Assessment and Development of Biomaterials-BioMat, Federal University of Vales do Jequitinhonha e Mucuri-UFVJM, Diamantina/MG 39100-000 (Brazil); Mansur, Herman S. [Department of Metallurgical and Materials Engineering, Federal University of Minas Gerais-UFMG, Belo Horizonte/MG 31270-901 (Brazil); Heneine, Luiz Guilherme D. [Department of Health Science, Ezequiel Dias Foundation-FUNED, Belo Horizonte/MG 30510-010 (Brazil)

    2013-03-01

    Bovine pericardium is widely used as a raw material in bioengineering as a source of collagen, a fundamental structural molecule. The physical, chemical, and biocompatibility characteristics of these natural fibers enable their broad use in several areas of the health sciences. For these applications, it is important to obtain collagen of the highest possible purity. The lack of a method to produce these pure biocompatible materials using simple and economically feasible techniques presents a major challenge to their production on an industrial scale. This study aimed to extract, purify, and characterize the type I collagen protein originating from bovine pericardium, considered to be an abundant tissue resource. The pericardium tissue was collected from male animals at slaughter age. Pieces of bovine pericardium were enzymatically digested, followed by a novel protocol developed for protein purification using ion-exchange chromatography. The material was extensively characterized by electrophoresis, scanning electron microscopy, energy dispersive X-ray spectroscopy, and infrared spectroscopy. The results showed a purified material with morphological properties and chemical functionalities compatible with type I collagen and similar to a highly purified commercial collagen. Thus, an innovative and relatively simple processing method was developed to extract and purify type I collagen from bovine tissue with potential applications as a biomaterial for regenerative tissue engineering. - Highlights: Black-Right-Pointing-Pointer Type I collagen was obtained from bovine pericardium, an abundant tissue resource. Black-Right-Pointing-Pointer A simple and feasible processing technique was developed to purify bovine collagen. Black-Right-Pointing-Pointer The appropriate process may be performed on industrial scale. Black-Right-Pointing-Pointer The pure collagen presented appropriate morphological and molecular characteristics. Black-Right-Pointing-Pointer The purify

  4. Prevalence of cysticercosis in Estonian pigs and cattle

    DEFF Research Database (Denmark)

    Dorbek-Kilin, Elisabeth; Åhlberg, Tuuli; Tummeleht, Lea

    2018-01-01

    Taenia solium has been ranked as the most important foodborne parasite and Taenia saginata as the most commonly found human Taenia tapeworm worldwide. The last official reports of taeniosis from Estonia were in 2003 for T. solium and 2012 for T. saginata. By law, all animal cases of cysticercosis...

  5. Screening of the Open Source Malaria Box Reveals an Early Lead Compound for the Treatment of Alveolar Echinococcosis.

    Directory of Open Access Journals (Sweden)

    Britta Stadelmann

    2016-03-01

    Full Text Available The metacestode (larval stage of the tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE, a very severe and in many cases incurable disease. To date, benzimidazoles such as albendazole and mebendazole are the only approved chemotherapeutical treatment options. Benzimidazoles inhibit metacestode proliferation, but do not act parasiticidal. Thus, benzimidazoles have to be taken a lifelong, can cause adverse side effects such as hepatotoxicity, and are ineffective in some patients. We here describe a newly developed screening cascade for the evaluation of the in vitro efficacy of new compounds that includes assessment of parasiticidal activity. The Malaria Box from Medicines for Malaria Venture (MMV, comprised of 400 commercially available chemicals that show in vitro activity against Plasmodium falciparum, was repurposed. Primary screening was carried out at 10 μM by employing the previously described PGI assay, and resulted in the identification of 24 compounds that caused physical damage in metacestodes. Seven out of these 24 drugs were also active at 1 μM. Dose-response assays revealed that only 2 compounds, namely MMV665807 and MMV665794, exhibited an EC50 value below 5 μM. Assessments using human foreskin fibroblasts and Reuber rat hepatoma cells showed that the salicylanilide MMV665807 was less toxic for these two mammalian cell lines than for metacestodes. The parasiticidal activity of MMV665807 was then confirmed using isolated germinal layer cell cultures as well as metacestode vesicles by employing viability assays, and its effect on metacestodes was morphologically evaluated by electron microscopy. However, both oral and intraperitoneal application of MMV665807 to mice experimentally infected with E. multilocularis metacestodes did not result in any reduction of the parasite load.

  6. Process for purifying molybdenum

    International Nuclear Information System (INIS)

    Cheresnowsky, J.

    1989-01-01

    This patent describes a process for purifying molybdenum containing arsenic and phosphorus. The process comprising: adding to an acidic slurry of molybdenum trioxide, a source of magnesium ions in a solid form, with the amount of magnesium and the magnesium ion concentration in the subsequently formed ammonium molybdate solution being sufficient to subsequently form insoluble compounds containing greater than about 80% by weight of the arsenic and greater than about 80% by weight of the phosphorus, and ammonia in an amount sufficient to subsequently dissolve the molybdenum and subsequently form the insoluble compounds, with the source of magnesium ions being added prior to the addition of the ammonia; digesting the resulting ammoniated slurry at a temperature sufficient to dissolve the molybdenum and form an ammonium molybdate solution while the pH is maintained at from bout 9 to about 10 to form a solid containing the insoluble compounds; and separating the solid from the ammonium molybdate solution

  7. Method of purifying phosphoric acid after solvent extraction

    International Nuclear Information System (INIS)

    Kouloheris, A.P.; Lefever, J.A.

    1979-01-01

    A method of purifying phosphoric acid after solvent extraction is described. The phosphoric acid is contacted with a sorbent which sorbs or takes up the residual amount of organic carrier and the phosphoric acid separated from the organic carrier-laden sorbent. The method is especially suitable for removing residual organic carrier from phosphoric acid after solvent extraction uranium recovery. (author)

  8. 78 FR 69361 - Development of Inward Leakage Standards for Half-Mask Air-Purifying Particulate Respirators

    Science.gov (United States)

    2013-11-19

    ... DEPARTMENT OF HEALTH AND HUMAN SERVICES 42 CFR Part 84 [Docket No. CDC-2013-0017; NIOSH-250] Development of Inward Leakage Standards for Half-Mask Air- Purifying Particulate Respirators AGENCY: Centers... regarding the development of inward leakage performance standards for half-mask air- purifying particulate...

  9. Characterization of a purified nicotinic receptor from rat brain by using idiotypic and anti-idiotypic antibodies

    International Nuclear Information System (INIS)

    Abood, L.G.; Langone, J.J.; Bjercke, R.; Lu, X.; Banerjee, S.

    1987-01-01

    The availability of an anti-nicotine monoclonal antibody has made it possible to further establish the nature of the nicotine recognition proteins purified from rat brain by affinity chromatography and to provide a highly sensitive assay for determining [ 3 H]nicotine binding to the purified material. An enantiomeric analogue of nicotine. (-)-6-hydroxymethylnicotine, was used to prepare the affinity column. In addition, with the use of an anti-idiotypic monoclonal antibody, it was confirmed that the recognition site for nicotine resides on a protein complex composed of two components with molecular masses of 62 and 57 kDa. It was also demonstrated that the same two proteins could be purified by immunoaffinity chromatography with the use of an anti-idiotypic monoclonal antibody. With the use of the anti-nicotine antibody to measure [ 3 H]nicotine binding, the purified material was shown to bind 250 pmol/mg of protein. By utilizing a procedure in which the purified receptor protein was conjugated to membranes by disulfide bonds, a binding activity of 80 pmol/mg was obtained. With the availability of sterospecific monoclonal antibodies to (-)-nicotine as well as monoclonal anti-idiotypic antibodies derived when the anti-nicotine antibodies were used as immunogens, additional procedures became available for the further characterization of the purified nicotine receptor and examining its (-)-[ 3 H]nicotine-binding characteristics

  10. Improved detection of a staphylococcal infection by monomeric and protein A-purified polyclonal human immunoglobulin

    International Nuclear Information System (INIS)

    Calame, W.

    1993-01-01

    The present study was undertaken to compare the technetium-99m labelled non-specific polyclonal human immunoglobulin (Ig) with 99m Tc-labelled monomeric human immunoglobulin (m-Ig), 99m Tc-labelled, protein A-purified, human immunoglobulin (A-IG) and 99m Tc-labelled monomeric, protein A-purified, human immunoglobulin (mA-Ig) as tracer agents for the detection of a thigh infection with Staphylococcus aureus. In vitro the binding of the various tracer agents to bacteria at various intervals was determined. For the in vivo evaluation, mice were infected and received one of the various labelled proteins. Scintigrams were made 0.25, 1, 4 and 24 h later. All 99m Tc-labelled Igs bound to bacteria in vitro: The percentages of binding for the m-Ig (from 1 h onwards) and A-Ig and mA-Ig (from 3 h onwards) were significantly higher than that for Ig. The in vivo target-to-non-target (T/NT) ratios were significantly higher from 4 h onwards for all purified Igs than for Ig. Protein A-purified Ig yielded higher T/NT ratios than m-Ig. Furthermore, the amount of activity in the liver was significantly lower 24 h after administration of m-Ig, A-Ig and mA-Ig than after administration of Ig. It is concluded that in this experimental infection 99m Tc-labelled monomeric Ig localizes a staphylococcal thigh infection better and faster than 99m Tc-labelled unpurified Ig. However, the accumulation obtained with protein A-purified Ig or protein A-purified monomeric Ig was the highest of all tracer agents tested. (orig.)

  11. Caspase inhibitors of the P35 family are more active when purified from yeast than bacteria.

    Directory of Open Access Journals (Sweden)

    Ingo L Brand

    Full Text Available Many insect viruses express caspase inhibitors of the P35 superfamily, which prevent defensive host apoptosis to enable viral propagation. The prototypical P35 family member, AcP35 from Autographa californica M nucleopolyhedrovirus, has been extensively studied. Bacterially purified AcP35 has been previously shown to inhibit caspases from insect, mammalian and nematode species. This inhibition occurs via a pseudosubstrate mechanism involving caspase-mediated cleavage of a "reactive site loop" within the P35 protein, which ultimately leaves cleaved P35 covalently bound to the caspase's active site. We observed that AcP35 purifed from Saccharomyces cerevisae inhibited caspase activity more efficiently than AcP35 purified from Escherichia coli. This differential potency was more dramatic for another P35 family member, MaviP35, which inhibited human caspase 3 almost 300-fold more potently when purified from yeast than bacteria. Biophysical assays revealed that MaviP35 proteins produced in bacteria and yeast had similar primary and secondary structures. However, bacterially produced MaviP35 possessed greater thermal stability and propensity to form higher order oligomers than its counterpart purified from yeast. Caspase 3 could process yeast-purified MaviP35, but failed to detectably cleave bacterially purified MaviP35. These data suggest that bacterially produced P35 proteins adopt subtly different conformations from their yeast-expressed counterparts, which hinder caspase access to the reactive site loop to reduce the potency of caspase inhibition, and promote aggregation. These data highlight the differential caspase inhibition by recombinant P35 proteins purified from different sources, and caution that analyses of bacterially produced P35 family members (and perhaps other types of proteins may underestimate their activity.

  12. Aspartic acid racemisation in purified elastin from arteries as basis for age estimation.

    Science.gov (United States)

    Dobberstein, R C; Tung, S-M; Ritz-Timme, S

    2010-07-01

    Aspartic acid racemisation (AAR) results in an age-dependent accumulation of D: -aspartic acid in durable human proteins and can be used as a basis for age estimation. Routinely, age estimation based on AAR is performed by analysis of dentine. However, in forensic practise, teeth are not always available. Non-dental tissues for age estimation may be suitable for age estimation based on AAR if they contain durable proteins that can be purified and analysed. Elastin is such a durable protein. To clarify if purified elastin from arteries is a suitable sample for biochemical age estimation, AAR was determined in purified elastin from arteries from individuals of known age (n = 68 individuals, including n = 15 putrefied corpses), considering the influence of different stages of atherosclerosis and putrefaction on the AAR values. AAR was found to increase with age. The relationship between AAR and age was good enough to serve as basis for age estimation, but worse than known from dentinal proteins. Intravital and post-mortem degradation of elastin may have a moderate effect on the AAR values. Age estimation based on AAR in purified elastin from arteries may be a valuable additional tool in the identification of unidentified cadavers, especially in cases where other methods cannot be applied (e.g., no available teeth and body parts).

  13. Synthesis and characterization of highly purified nanosilica from pyrophyllite ores

    Energy Technology Data Exchange (ETDEWEB)

    Fuad, Abdulloh, E-mail: abdulloh.fuad.fmipa@um.ac.id; Mufti, Nandang; Diantoro, Markus; Subakti,; Septa Kurniawati, S. [Jurusan Fisika FMIPA Universitas Negeri Malang. Jl. Semarang No. 5 Malang, east Java (Indonesia)

    2016-03-11

    A simple method based on alkaline extraction followed by acid precipitation and acid dissolution has been developed to produce highly purified nanosilica from pyrophyllite materials. The reaction parameters such as molar ratio NaOH/SiO{sub 2}, reaction time and reaction temperature are varied for obtaining maximum nanosilica convertion. About 99,45% highly purified precipitated nanosilica measure with ICP, 259 m{sup 2}/gr measure with BET surface area, 97% whiteness and 3 ml/gr oil absorbtion from pyrophyllite materials has been achieved in closed system at 150°C within 180 min. The physicals and chemical properties (such as X-Ray Diffraction from PANalytical, X-Ray Fluorescence Minipal4 from PANanalytical, BET surface area, Forier Transform Infra Red Spectroscopy from Hitachi, and SEM-EDS Inspect-S50 from FEI Company) of the highly purity nanosilica were studied.

  14. Affinity-purified human interleukin I is cytotoxic to isolated islets of Langerhans

    DEFF Research Database (Denmark)

    Mandrup-Poulsen, T; Bendtzen, K; Nerup, J

    1986-01-01

    Addition of highly purified human Interleukin-1 to the culture medium of isolated rat islets of Langerhans for 6 days led to 88% inhibition of glucose-induced insulin-release, reduction of islet contents of insulin and glucagon to 31% and 8% respectively, and disintegration of the islets. These e......Addition of highly purified human Interleukin-1 to the culture medium of isolated rat islets of Langerhans for 6 days led to 88% inhibition of glucose-induced insulin-release, reduction of islet contents of insulin and glucagon to 31% and 8% respectively, and disintegration of the islets...

  15. The alpaca (Vicugna pacos) as a natural intermediate host of Taenia omissa (Cestoda: Taeniidae).

    Science.gov (United States)

    Gomez-Puerta, Luis A; Yucra, Dora; Lopez-Urbina, Maria T; Gonzalez, Armando E

    2017-11-15

    Three metacestodes were collected from the mesentery and the surface of the liver of three adult alpacas (Vicugna pacos) in a slaughterhouse located in Puno, Peru. Various features of the metacestodes were observed for morphological identification. A molecular diagnosis was performed by PCR-based sequencing of mitochondrial genes of cytochrome c oxidase subunit 1 (cox1) and the NADH dehydrogenase subunit 1 (nad1). All metacestodes were identified as Taenia omissa by morphology and molecular methods The isolates from alpacas showed significant sequence similarity with previously reported isolates of T. omissa (95.7-98.1% in cox1 and 94.6-95.1% in nad1). Our report is the first to detect T. omissa metacestodes in alpacas and to reveal that alpacas are natural intermediate hosts for this parasite. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Immunomodulatory activity of purified arabinoxylans from finger millet (Eleusine coracana, v. Indaf 15) bran.

    Science.gov (United States)

    Savitha Prashanth, M R; Shruthi, R R; Muralikrishna, G

    2015-09-01

    Biological activities of alkali extracted (Barium hydroxide: BE-480 kDa, Potassium hydroxide: KE1-1080 and KE2-40 kDa), purified arabinoxylans (AX) from the finger millet bran varying in their molecular weight, phenolic acid content, arabinose to xylose ratios were evaluated for their immune-stimulatory activities using murine lymphocytes and peritoneal exudate macrophages. All three purified AX displayed significant (p 2 fold) and macrophage phagocytosis than KE1 and KE2. The above results clearly documented that the immunostimulatory activity of arabinoxylans is directly proportional to the amount of ferulic acid content (0.11 mg/100 g), whereas molecular weight as well as arabinose/xylose ratio, did not have any bearing. Purified AX from the finger millet bran can be explored as a potent natural immunomodulator.

  17. The Asian Taenia and the possibility of cysticercosis

    Science.gov (United States)

    V. Fuentes, Màrius

    2000-01-01

    In certain Asian countries, a third form of human Taenia, also known as the Asian Taenia, has been discovered. This Asian Taenia seems to be an intermediate between Taenia solium and T. saginata since in morphological terms it is similar to T. saginata, yet biologically, as it uses the same intermediate host (pigs), it is more akin to T. solium. Taenia solium causes human cysticercosis, while T. saginata does not. It is not known whether the Asian taeniid is able to develop to the larval stage in humans or not. The arguments proposed by those authors who consider it unlikely that the Asian Taenia causes human cysticercosis are: (a) its molecular similarities with T. saginata; (b) the absence of cases of human cysticercosis in populations where the Asian adult is highly prevalent; and (c) the unsupporting results derived from an experimental infestation study. These three arguments are debated, although bearing in mind that at present there is still no clear scientific data to support that human cysticercosis can be caused by the Asian Taenia. PMID:10743352

  18. Activation of purified calcium channels by stoichiometric protein phosphorylation

    Energy Technology Data Exchange (ETDEWEB)

    Nunoki, K.; Florio, V.; Catterall, W.A. (Univ. of Washington, Seattle (USA))

    1989-09-01

    Purified dihydropyridine-sensitive calcium channels from rabbit skeletal muscle were reconstituted into phosphatidylcholine vesicles to evaluate the effect of phosphorylation by cyclic AMP-dependent protein kinase (PK-A) on their function. Both the rate and extent of {sup 45}Ca{sup 2+} uptake into vesicles containing reconstituted calcium channels were increased severalfold after incubation with ATP and PK-A. The degree of stimulation of {sup 45}Ca{sup 2+} uptake was linearly proportional to the extent of phosphorylation of the alpha 1 and beta subunits of the calcium channel up to a stoichiometry of approximately 1 mol of phosphate incorporated into each subunit. The calcium channels activated by phosphorylation were determined to be incorporated into the reconstituted vesicles in the inside-out orientation and were completely inhibited by low concentrations of dihydropyridines, phenylalkylamines, Cd{sup 2+}, Ni{sup 2+}, and Mg{sup 2+}. The results demonstrate a direct relationship between PK-A-catalyzed phosphorylation of the alpha 1 and beta subunits of the purified calcium channel and activation of the ion conductance activity of the dihydropyridine-sensitive calcium channels.

  19. Activation of purified calcium channels by stoichiometric protein phosphorylation

    International Nuclear Information System (INIS)

    Nunoki, K.; Florio, V.; Catterall, W.A.

    1989-01-01

    Purified dihydropyridine-sensitive calcium channels from rabbit skeletal muscle were reconstituted into phosphatidylcholine vesicles to evaluate the effect of phosphorylation by cyclic AMP-dependent protein kinase (PK-A) on their function. Both the rate and extent of 45 Ca 2+ uptake into vesicles containing reconstituted calcium channels were increased severalfold after incubation with ATP and PK-A. The degree of stimulation of 45 Ca 2+ uptake was linearly proportional to the extent of phosphorylation of the alpha 1 and beta subunits of the calcium channel up to a stoichiometry of approximately 1 mol of phosphate incorporated into each subunit. The calcium channels activated by phosphorylation were determined to be incorporated into the reconstituted vesicles in the inside-out orientation and were completely inhibited by low concentrations of dihydropyridines, phenylalkylamines, Cd 2+ , Ni 2+ , and Mg 2+ . The results demonstrate a direct relationship between PK-A-catalyzed phosphorylation of the alpha 1 and beta subunits of the purified calcium channel and activation of the ion conductance activity of the dihydropyridine-sensitive calcium channels

  20. Influence of a highly purified senna extract on colonic epithelium

    NARCIS (Netherlands)

    van Gorkom, B A; Karrenbeld, A; van Der Sluis, T; Koudstaal, J; de Vries, E G; Kleibeuker, J H

    2000-01-01

    BACKGROUND: Chronic use of sennoside laxatives often causes pseudomelanosis coli. A recent study suggested that pseudomelanosis coli is associated with an increased colorectal cancer risk. A single high dose of highly purified senna extract increased proliferation rate and reduced crypt length in

  1. Characterization of product RNAs synthesized in vitro by poliovirus RNA polymerase purified by chromatography on hydroxylapatite or poly(U) Sepharose.

    OpenAIRE

    Young, D C; Tobin, G J; Flanegan, J B

    1987-01-01

    The size of the product RNA synthesized by the poliovirus RNA polymerase and host factor was significantly affected by the type of column chromatography used to purify the polymerase. Dimer length product RNA was synthesized by the polymerase purified by chromatography on hydroxylapatite. This contrasted with the monomer length product RNA synthesized by the polymerase purified by chromatography on poly(U) Sepharose. The poly(U) Sepharose-purified polymerase was shown to contain oligo(U) that...

  2. Oxidative Stability of Dispersions Prepared from Purified Marine Phospholipid and the Role of α-Tocopherol

    DEFF Research Database (Denmark)

    Lu, Henna Fung Sieng; Nielsen, Nina Skall; Baron, Caroline P.

    2012-01-01

    , respectively, during 32 days of storage at 2 °C. Nonenzymatic browning was investigated through measurement of Strecker aldehydes, color changes, and pyrrole content. Dispersions containing α-tocopherol or higher levels of purified marine PL showed a lower increment of volatiles after 32 days storage......The objective of this study was to investigate the oxidative stability of dispersions prepared from different levels of purified marine phospholipid (PL) obtained by acetone precipitation, with particular focus on the interaction between α-tocopherol and PL in dispersions. This also included...... the investigation of nonenzymatic browning in purified marine PL dispersions. Dispersions were prepared by high-pressure homogenizer. The oxidative and hydrolytic stabilities of dispersions were investigated by determination of hydroperoxides, secondary volatile oxidation products, and free fatty acids...

  3. Air purification by cementitious materials: Evaluation of air purifying properties

    NARCIS (Netherlands)

    Hüsken, G.; Brouwers, H.J.H.; Al-Mattarneh, H.; Mustapha, K.N.; Nuruddin, M.F.

    2008-01-01

    This paper addresses the evaluation of the photocatalytic properties of concrete containing titanium dioxide (TiO2). Here, the assessment of the air purifying abilities of the hardened concrete regarding the degradation of nitric oxide (NO) is of major interest. A setup for measuring the performance

  4. Air purification by cementitious materials : Evaluation of air purifying properties

    NARCIS (Netherlands)

    Hüsken, G.; Brouwers, H.J.H.; Al-Mattarneh, H.; Mustapha, K.N.; Nuruddin, M.F.

    2008-01-01

    This paper addresses the evaluation of the photocatalytic properties of concrete containing titanium dioxide (TiO2). Here, the assessment of the air purifying abilities of the hardened concrete regarding the degradation of nitric oxide (NO) is of major interest. A setup for measuring the performance

  5. High-level water purifying technology. Kodo josui shori gijutsu

    Energy Technology Data Exchange (ETDEWEB)

    Tsugura, H; Tsukiashi, K [Meidensha Corp., Tokyo (Japan)

    1993-07-01

    Research and development have been carried out on a high-level water purifying system using ozone and activated charcoals to supply drinking water free of carcinogenic matters and odors. This system comprises a system to utilize ozone by using silent discharge and oxygen enriching device, and a living organism/activated charcoal treatment system. The latter system utilizes living organisms deposited on activated charcoal surfaces to remove polluting substances including ammonia. The treatment experimenting equipment comprises an ozone generating system, an ozone treating column, an activated charcoal treating column, an ozone/activated charcoal control device, and a water amount and quality measuring system. An experiment was carried out using an experimental plant with a capacity of 20 m[sup 3]/day on water taken from the sedimentation process at an actual water purifying plant. As a result, trihalomethane formation potential was removed at about 40% in the ozone treatment, and at 70% in the whole treatment combining the ozone and living organism/activated charcoal treatments. For parameterization of palatability of water, a method is being studied that utilizes nuclear magnetic resonance to investigate degrees of water cluster. The method is regarded promising. 1 ref., 4 figs.

  6. Biological and Histological Studies of Purified Product from Streptomyces janthinus M7 Metabolites

    Directory of Open Access Journals (Sweden)

    Tawfik Zahira S.

    2015-02-01

    Full Text Available Fifteen clinical samples were taken out from patients suffering cancer, these patients being under the treatment with radio- and/or chemotherapy. The samples were used for the isolation of bacterial cells surrounding tumor; the samples were collected from Center of Cancer Therapy, Ain Shams University, Cairo, Egypt. The clinical bacterial isolates were purified and identified according to Bergey's manual of determinative bacteriology ninth edition (1994. The bacterial isolates were found to be Klebsiella oxytoca m1; Enterobacter cancerogenus m2; P. aeruginosa m3; Citrobacter diversus m4; Enterobacter agglomerans m5; Klebsiella oxytoca m6; Enterobacter dissolvens m7; Serratia fonticola m8; Escherichia coli m9; Citrobacter freundii m10; Staphylococcus aureus m11; Escherichia coli m12; P. aeruginosa m13; Staphylococcus aureus m14; and Bacillus cereus m15. In the present study both primary and secondary screening methods were used to screen the antibacterial activity of St. janthinus M7 against fifteen clinical bacterial isolates. The St. janthinus M7 showed an increase in antibacterial activity against all the tested human bacterial pathogens. In this study Gamma irradiation at dose levels (0.5 and 1.5 kGy was used for the enhancement of the antibacterial activity of Streptomyces strain against the clinical isolates. Several commercial antibiotic discs (Doxorubicin, Augmentin, Norfloxacin, Ofloxacin, Oxacillin, and Cefazolin were used for comparing their antimicrobial activity with purified product. The results declared a significant increase in the antibacterial activity in most cases. The physiochemical properties of the purified product were carried out for determination of Rf, empirical formula, M.W, and chemical structure of product and then analyzed by thin layer chromatography, elemental analysis, UV, Mass, and NMR. The result exhibited brown color, one spot, Rf (0.76, M.W (473, while it recorded 270 nm in UV region and the calculated

  7. Taeniases and cysticercosis in Indonesia: past and present situations.

    Science.gov (United States)

    Wandra, Toni; Ito, Akira; Swastika, Kadek; Dharmawan, Nyoman S; Sako, Yasuhito; Okamoto, Munehiro

    2013-11-01

    The main aim of this study is to overview the past and present situations of human taeniases and cysticercosis in Indonesia and including future perspectives. Through joint projects from 1996, we have confirmed the occurrence of Taenia saginata (beef tapeworm) in Bali, of Taenia solium (pork tapeworm) mainly in Papua and sporadically in Bali, and of Taenia asiatica in North Sumatra. These taeniases were caused through eating uncooked pork and pig viscera for T. solium and T. asiatica, respectively, and beef for T. saginata. The distribution of these tapeworms in Indonesia is basically highly restricted by the traditional cultural and religious backgrounds in each island. T. saginata is relatively common in Bali although people consume pork 'lawar' more than beef 'lawar'. Taeniases due to T. saginata or T. asiatica and T. solium and cysticercosis due to T. solium have also been sporadically reported in some other islands. Among these species, T. solium is exceptional since humans can be infected not only by larval stages (cysticerci) in pork but also by eggs released from human tapeworm carriers. Cysticercosis has been confirmed in Indonesia in humans, pigs and even dogs.

  8. Taeniasis and cysticercosis in Asia and the Pacific: present state of knowledge and perspectives.

    Science.gov (United States)

    Ito, Akira; Nakao, Minoru; Wandra, Toni; Suroso, Thomas; Okamoto, Munehiro; Yamasaki, Hiroshi; Sako, Yasuhito; Nakaya, Kazuhiro

    2005-01-01

    Several topics on taeniasis and cysticercosis in Asia and the Pacific are overviewed. In Asia and the Pacific, three human taeniid species have been recognized: Taenia solium, Taenia saginata and Taenia asiatica. The first topic is on evolution of T. solium. Mitochondrial DNA polymorphisms of T. solium worldwide are discussed with emphasis of two specific genotypes: American-African and Asian. The second topic is recent major advances in sero- and molecular-diagnosis of T. solium cysticercosis in humans, pigs and dogs. The third is the present situation of T. solium taeniasis/cysticercosis in Papua (Irian Jaya), Indonesia. The forth is the present situation of T. solium cysticercosis and T. saginata taeniasis in Bali, Indonesia. The fifth is the present situation of T. asiatica taeniasis in Asia and the Pacific and in North Sumatra, Indonesia. The sixth is on the debate of the exact definition of T. asiatica. Because T. asiatica can not be differentiated from T. saginata morphologically, it is time to re-evaluate T. saginata in Asia and the Pacific. New and broad-based surveys across this region are necessary from epidemiological and public health perspectives, based on evidence.

  9. 76 FR 3159 - Purified Carboxymethylcellulose From Finland, Mexico, Netherlands, and Sweden

    Science.gov (United States)

    2011-01-19

    ... INTERNATIONAL TRADE COMMISSION [Investigation No. 731-TA-1084-1087 (Review)] Purified Carboxymethylcellulose From Finland, Mexico, Netherlands, and Sweden AGENCY: United States International Trade Commission. ACTION: Revised schedule for the subject reviews. DATES: Effective Date: January 7, 2011. FOR FURTHER...

  10. AQUAPEAT 95. New methods for purifying the run-offs of peat production areas

    International Nuclear Information System (INIS)

    Selin, P.; Marja-aho, J.; Madekivi, O.

    1994-01-01

    The aim of Aqua Peat 95-project was to develop new methods for purifying the runoff coming from the peat production areas. The national water protection program for the year 1995 (Ympaeristoeministerioe 1988) as well as the level of the requirements and instructions from the authorities will obligate the peat producers to find new and practical methods for water purification. The chemical treatment reduced the load of peat production areas and the quality of treated water was almost equal to the runoffs coming from the natural bog area. The chemicals were the same as used in purifying drinking water. This purifying method is quite expensive and for this reason applicable only in special cases. The transpiration and evaporation and the soil filtering capacity of the forest area was also observed. The purifying capacity was very good, especially for the total nutrients and suspended solids. The changes of the ground water quality were insignificant but the level of the ground water in the field areas was higher than before. The long term changes of the vegetation and the trees could not be seen, yet. The most important water management practice is the detention of the discharge. The capacity of the sedimentation will increase by using the flow regulation in the sedimentation ponds and ditches. The changes in the water biology downstreams the Laeynioensuo peat production area were clearly seen near the main ditch. Because of the suspended solids the bottom sediment changed which lead to impacts to the bottom fauna. The colour of the runoffs as well as the changes in the sediment influenced on the macrophytes

  11. The design and commissioning of cold trap purifying system of hydrogen meter sodium loop

    International Nuclear Information System (INIS)

    Zhao Zhaoyi; Jia Baoshan; Chen Xiaoming; Pan Fengguo

    1993-01-01

    The design feature and parameters of cold trap purifying system of hydrogen meter sodium loop and its commissioning results are reported and discussed. In order to adjust the flow easily,. the cold trap purifying system is arranged in the exit of the electromagnetic pump. It is composed of regenerator and the cold trap. The regenerator is above the cold trap. The high temperature sodium in the main-loop flows through the regenerator, in the entrance of the cold trap, its temperature is reduced to 180 degree C. After entering into the cold trap, the sodium flows to the purifying region by side, when it arrives the bottom of the trap, its temperature is reduced to 110 degree C. The cold trap is cooled by air. The temperature of the clean sodium rises nearby the main-loop's by the regenerator, and then it returns to the entrance of the electromagnetic pump. According to the commissioning results, the sodium's temperature of the cold trap could be reduced to 110 degree C by reducing the flow of the cold trap purifying system and the temperature of the main-loop, or increasing the air flow and cutting off the power supply of its heating. The authors think that the latter is more conformable with the design stipulation and with the requirement of the hydrogen meter experiment, and it can meet the requirements of the operation of the Nuclear Power Plant

  12. Integrated Microchannel Reformer/Hydrogen Purifier for Fuel Cell Power Systems, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — Makel Engineering, Inc. (MEI) and Colorado School of Mines (CSM) propose to develop an integrated hydrogen generator and purifier system for conversion of in-situ...

  13. Consumer Behavior Modeling: Fuzzy Logic Model for Air Purifiers Choosing

    Directory of Open Access Journals (Sweden)

    Oleksandr Dorokhov

    2017-12-01

    Full Text Available At the beginning, the article briefly describes the features of the marketing complex household goods. Also provides an overview of some aspects of the market for indoor air purifiers. The specific subject of the study was the process of consumer choice of household appliances for cleaning air in living quarters. The aim of the study was to substantiate and develop a computer model for evaluating by the potential buyers devices for air purification in conditions of vagueness and ambiguity of their consumer preferences. Accordingly, the main consumer criteria are identified, substantiated and described when buyers choose air purifiers. As methods of research, approaches based on fuzzy logic, fuzzy sets theory and fuzzy modeling were chosen. It was hypothesized that the fuzzy-multiple model allows rather accurately reflect consumer preferences and potential consumer choice in conditions of insufficient and undetermined information. Further, a computer model for estimating the consumer qualities of air cleaners by customers is developed. A proposed approach based on the application of fuzzy logic theory and practical modeling in the specialized computer software MATLAB. In this model, the necessary membership functions and their terms are constructed, as well as a set of rules for fuzzy inference to make decisions on the estimation of a specific air purifier. A numerical example of a comparative evaluation of air cleaners presented on the Ukrainian market is made and is given. Numerical simulation results confirmed the applicability of the proposed approach and the correctness of the hypothesis advanced about the possibility of modeling consumer behavior using fuzzy logic. The analysis of the obtained results is carried out and the prospects of application, development, and improvement of the developed model and the proposed approach are determined.

  14. Synthesis and characterization of nano-sized CaCO3 in purified diet

    Science.gov (United States)

    Mulyaningsih, N. N.; Tresnasari, D. R.; Ramahwati, M. R.; Juwono, A. L.; Soejoko, D. S.; Astuti, D. A.

    2017-07-01

    The growth and development of animals depend strongly on the balanced nutrition in the diet. This research aims is to characterize the weight variations of nano-sized calcium carbonate (CaCO3) in purified diet that to be fed to animal model of rat. The nano-sized CaCO3 was prepared by milling the calcium carbonate particles for 20 hours at a rotation speed of 1000 rpm and resulting particle size in a range of 2-50 nm. Nano-sized CaCO3 added to purified diet to the four formulas that were identified as normal diet (N), deficiency calcium (DC), rich in calcium (RC), and poor calcium (PC) with containing in nano-sized CaCO3 much as 0.50 %, 0.00 %, 0.75 % and 0.25 % respectively. The nutritional content of the purified diet was proximate analyzed, it resulted as followed moisture, ash, fat, protein, crude fiber. The quantities of chemical element were analyzed by atomic absorption spectrometry (AAS), it resulted iron, magnesium, potassium and calcium. The results showed that N diet (Ca: 16,914.29 ppm) were suggested for healthy rats and RC diet (Ca: 33,696.13 ppm) for conditioned osteoporosis rats. The crystalline phases of the samples that were examined by X-ray diffraction showed that crystalline phase increased with the increasing concentration of CaCO3.

  15. Endurance Pump Tests With Fresh and Purified MIL-PRF-83282 Hydraulic Fluid

    National Research Council Canada - National Science Library

    Sharma, Shashi

    1999-01-01

    .... Two endurance pump tests were conducted with F-16 aircraft hydraulic pumps, using both fresh and purified MIL-PRF-83282 hydraulic fluid, to determine if fluid purification had any adverse effect on pump life...

  16. Occurrence of Conjugated Linolenic Acids in Purified Soybean Oil

    OpenAIRE

    Kinami, Tomohisa; Horii, Naoto; Narayan, Bhaskar; Arato, Shingo; Hosokawa, Masashi; Miyashita, Kazuo; Negishi, Hironori; Ikuina, Junichi; Noda, Ryuji; Shirasawa, Seiichi

    2007-01-01

    A high-performance liquid chromatographic (HPLC) method is described for the determination of conjugated linoleic acids (CLA) and conjugated linolenic acids (CLN). Methyl esters prepared from purified lipid fractions of soybean oil were analyzed using an HPLC system equipped with photodiode-array detector to detect peaks having maximum absorption around 233 and 275 nm. These peaks were concentrated by AgNO3-silicic acid column chromatography and reversed-phase HPLC. The structural analysis, o...

  17. Inference of purifying and positive selection in three subspecies of chimpanzees (Pan troglodytes) from exome sequencing

    DEFF Research Database (Denmark)

    Bataillon, Thomas; Duan, Jinjie; Hvilsom, Christina

    2015-01-01

    of recent gene flow from Western into Eastern chimpanzees. The striking contrast in X-linked vs. autosomal polymorphism and divergence previously reported in Central chimpanzees is also found in Eastern and Western chimpanzees. We show that the direction of selection (DoS) statistic exhibits a strong non......-monotonic relationship with the strength of purifying selection S, making it inappropriate for estimating S. We instead use counts in synonymous vs. non-synonymous frequency classes to infer the distribution of S coefficients acting on non-synonymous mutations in each subspecies. The strength of purifying selection we...... infer is congruent with the differences in effective sizes of each subspecies: Central chimpanzees are undergoing the strongest purifying selection followed by Eastern and Western chimpanzees. Coding indels show stronger selection against indels changing the reading frame than observed in human...

  18. Fabrication of Simple Indoor Air Haze Purifier using Domestic Discarded Substances and Its Haze Removal Performance

    Science.gov (United States)

    Wang, Zhou; Cao, Haoshu; Zhao, Shuang

    2018-01-01

    Based on the concept of circular economy, discarded plastic bottles stuffed with discarded cotton, clothing and sofa cushion were used as pre-filter to remove big particles (dust and coal dust) in air and 4 L tap water in discarded plastic bottle was worked as an absorbing medium to dissolve the water soluble ions in air (SO4 2-, NO3-, NH4+, Cl- and Ca2+). Moreover, the internet control design was used in this homemade indoor air haze purifier to achieve the performance of remote control and intelligent management. The experimental results showed that this indoor air haze purifier can effectively reduce the level of indoor air haze and the air quality after 20 minutes treatment is higher than that of two commercial well-known air haze purifier

  19. Detection of taeniid (Taenia spp., Echinococcus spp. eggs contaminating vegetables and fruits sold in European markets and the risk for metacestode infections in captive primates

    Directory of Open Access Journals (Sweden)

    Karin Federer

    2016-12-01

    Full Text Available Due to frequent cases of alveolar echinococcosis (AE in captive primates in Europe, 141 samples of food, which consisting of vegetables and fruits, were investigated for contamination with egg-DNA of taeniids. Each sample consisted of at least 40 heads of lettuce as well as various vegetables and fruits. The samples were purchased at different times of the year: either from September to November (autumn, originating from greenhouses or fields in the Basel region in the North of Switzerland, or in April and May (spring when fruit and vegetables are sourced from throughout Europe from various wholesalers. Each sample was washed, and the washing water sieved through mesh apertures of 50 μm and 21 μm, respectively. The debris, including taeniid eggs, collected on the 21 μm sieve were investigated by a multiplex PCR-analysis followed by direct sequencing. In 17 (18% of the 95 samples collected in autumn, taeniid-DNA was detected (Taenia hydatigena in four, Taenia ovis in three, Taenia polyacantha in two and Hydatigera (Taenia taeniaeformis in five cases. Similarly, in 13 (28% of the 46 samples collected during spring taeniid-DNA was detected (Echinococcus granulosus s.l. in two, Taenia crassiceps in one, T. hydatigena in two, Taenia multiceps/Taenia serialis in two, Taenia saginata in one and H. taeniaeformis in five cases. Although DNA of Echinococcus multilocularis was not found specifically in this study, the detection of other fox taeniids reveals that vegetables and fruit fed to the primates at the Zoo Basel at different times of the year and from different origin are contaminated with carnivore's faeces and therefore act as a potential source of AE infections.

  20. Biocontrol activity of surfactin A purified from Bacillus NH-100 and NH-217 against rice bakanae disease.

    Science.gov (United States)

    Sarwar, Ambrin; Hassan, Muhammad Nadeem; Imran, Muhammad; Iqbal, Mazhar; Majeed, Saima; Brader, Günter; Sessitsch, Angela; Hafeez, Fauzia Yusuf

    2018-04-01

    The potential of the Bacillus genus to antagonize phytopathogens is associated with the production of cyclic lipopeptides. Depending upon the type of lipopeptide, they may serve as biocontrol agents that are eco-friendly alternatives to chemical fertilizers. This study evaluates the biocontrol activity of surfactin-producing Bacillus (SPB) strains NH-100 and NH-217 and purified surfactin A from these strains against rice bakanae disease. Biologically active surfactin fractions were purified by HPLC, and surfactin A variants with chain lengths from C12 to C16 were confirmed by LCMS-ESI. In hemolytic assays, a positive correlation between surfactin A production and halo zone formation was observed. The purified surfactin A had strong antifungal activity against Fusarium oxysporum, F. moniliforme, F. solani, Trichoderma atroviride and T. reesei. Maximum fungal growth suppression (84%) was recorded at 2000 ppm against F. moniliforme. Surfactin A retained antifungal activity at different pH levels (5-9) and temperatures (20, 50 and 121 °C). Hydroponic and pot experiments were conducted to determine the biocontrol activity of SPB strains and the purified surfactin A from these strains on Super Basmati rice. Surfactin production in the rice rhizosphere was detected by LCMS-ESI at early growth stages in hydroponics experiments inoculated with SPB strains. However, the maximum yield was observed with a consortium of SPB strains (T4) and purified surfactin A (T5) treatments in the pot experiment. The outcomes of the present study revealed that surfactin A significantly reduced rice bakanae disease by up to 80%. These findings suggest that purified surfactin A could be an effective biocontrol agent against bakanae disease in rice and should be incorporated into strategies for disease management. Copyright © 2018 Elsevier GmbH. All rights reserved.

  1. [Investigation on the process of sapindus saponin purified with macroporous adsorption resin and screening of its bacteriostasis].

    Science.gov (United States)

    Fu, Yong; Lei, Peng; Han, Yu-mei; Yan, Dan

    2010-02-01

    To study the technological parameters of the purification process of saponins with macroporous adsorption resin. The adsorptive characteristics and elutive parameters of the process were studied by taking the elutive and purified ratio of saponins as markers. Bacteriostasis activity of each parts eluted was evaluated by the mean of cup-plate method. 13.6 mL of the extraction of sapindus saponin (crude drugs 0.01 g/mL) was purified with a column of macroporous adsorption resin (phi15 mm x H90 mm, dry weight 2.5 g) and washed with 3BV of distilled water, then eluted with 3BV of 30% ethanol and 3BV of 70% ethanol, most of saponins were collected in the 70% ethanol. With macroporous adsorption resin adsorbing and purifying, the elutive ratio of saponins was 93.8% and the purity reached 250.1%. So this process of applying macroporous adsorption resin to adsorb and purify saponins is feasible, and supplies reference to the purification of other types of saponin.

  2. Apparatus and methods for purifying lead

    Science.gov (United States)

    Tunison, Harmon M.

    2016-01-12

    Disclosed is an exemplary method of purifying lead which includes the steps of placing lead and a fluoride salt blend in a container; forming a first fluid of molten lead at a first temperature; forming a second fluid of the molten fluoride salt blend at a second temperature higher than the first temperature; mixing the first fluid and the second fluid together; separating the two fluids; solidifying the molten fluoride salt blend at a temperature above a melting point of the lead; and removing the molten lead from the container. In certain exemplary methods the molten lead is removed from the container by decanting. In still other exemplary methods the molten salt blend is a Lewis base fluoride eutectic salt blend, and in yet other exemplary methods the molten salt blend contains sodium fluoride, lithium fluoride, and potassium fluoride.

  3. Evaluating the Effectiveness of a Commercial Portable Air Purifier in Homes with Wood Burning Stoves: A Preliminary Study

    Directory of Open Access Journals (Sweden)

    Julie F. Hart

    2011-01-01

    Full Text Available Wood burning for residential heating is prevalent in the Rocky Mountain regions of the United States. Studies have shown that wood stoves can be a significant source of PM2.5 within homes. In this study, the effectiveness of an electrostatic filter portable air purifier was evaluated (1 in a home where a wood stove was the sole heat source and (2 in a home where a wood stove was used as a supplemental heat source. Particle count concentrations in six particle sizes and particle mass concentrations in two particle sizes were measured for ten 12-hour purifier on and ten purifier off trials in each home. Particle count concentrations were reduced by 61–85 percent. Similar reductions were observed in particle mass concentrations. These findings, although limited to one season, suggest that a portable air purifier may effectively reduce indoor particulate matter concentrations associated with wood combustion during home heating.

  4. Directed growth of graphene nanomesh in purified argon via chemical vapor deposition.

    Science.gov (United States)

    Sun, Haibin; Fu, Can; Shen, Xia; Yang, Wenchao; Guo, Pengfei; Lu, Yang; Luo, Yongsong; Yu, Benhai; Wang, Xiaoge; Wang, Chunlei; Xu, Junqi; Liu, Jiangfeng; Song, Fengqi; Wang, Guanghou; Wan, Jianguo

    2017-06-16

    Graphene nanomeshes (GNMs), new graphene nanostructures with tunable bandgaps, are potential building blocks for future electronic or photonic devices, and energy storage and conversion materials. In previous works, GNMs have been successfully prepared on Cu foils by the H 2 etching effect. In this paper, we investigated the effect of Ar on the preparation of GNMs, and how the mean density and shape of them vary with growth time. In addition, scanning electron microscopy (SEM) and high resolution transmission electron microscopy (TEM) revealed the typical hexagonal structure of GNM. Atomic force microscopy (AFM) and x-ray photoelectron spectroscopy (XPS) indicated that large copper oxide nanoparticles produced by oxidization in purified Ar can play an essential catalytic role in preparing GNMs. Then, we exhibited the key reaction details for each growth process and proposed a growth mechanism of GNMs in purified Ar.

  5. The effect of a photocatalytic air purifier on indoor air quality quantified using different measuring methods

    DEFF Research Database (Denmark)

    Kolarik, Barbara; Wargocki, Pawel; Skorek-Osikowska, A.

    2010-01-01

    The effect on indoor air quality of an air purifier based on photocatalytic oxidation (PCO) was determined by different measuring techniques: sensory assessments of air quality made by human subjects, Proton-Transfer-Reaction Mass Spectrometry (PTR-MS) and chromatographic methods (Gas......, additional measurements were made with no pollution sources present in the office. All conditions were tested with the photocatalytic air purifier turned on and off. The results show that operation of the air purifier in the presence of pollutants emitted by building materials and furniture improves indoor...... Chromatography/Mass Spectrometry and High-Pressure Liquid Chromatography with UV detection). The experiment was conducted in a simulated office, ventilated with 0.6 h(-1), 2.5 h(-1) and 6 h(-1), in the presence of additional pollution sources (carpet, chipboard and linoleum). At the lowest air change rate...

  6. Creating and purifying an observation instrument using the generalizability theory

    Directory of Open Access Journals (Sweden)

    Elena Rodríguez-Naveiras

    2013-12-01

    Full Text Available The control of quality of data it is one of the most relevant aspects in observational researches. The Generalizability Theory (GT provides a method of analysis that allows us to isolate the various sources of error measurement. At the same time, it helps us to determine the extent to which various factors can change and analyze the effect on the generalizability coefficient. In the work shown here, there are two studies aimed to creating and purifying an observation instrument, Observation Protocol in the Teaching Functions (Protocolo de Funciones Docentes, PROFUNDO, v1 and v2, for behavioral assessment which has been carried out by instructors in a social-affective out-of-school program. The reliability and homogeneity studies are carried out once the instrument has been created and purified. The reliability study will be done through the GT method taking both codes (c and agents (a as differential facets in. The generalization will be done through observers using a crossed multi-faceted design (A × O × C. In the homogeneity study the generalization facet will be done through codes using the same design that the reliability study.

  7. Composition and potency characterization of Mycobacterium avium subsp. paratuberculosis purified protein derivatives

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) purified protein derivatives (PPDs) are immunologic reagents prepared from cultured filtrates of the type strain ATCC 19698. Traditional production consists of floating culture incubation at 37oC, organism inactivation by autoclaving, coarse filtrat...

  8. Worldwide Report, Epidemiology

    Science.gov (United States)

    1985-11-22

    caused by ingesting eggs of the pork tapeworm, Taenia solium . It has been described as the most com- mon parasitic disease of the central nervous...rather difficult to detect in stool specimens and at this stage it is impossible to distinguish between T. solium and T. saginata eggs either by...Knowledge and Perspectives, pp. 25-38. Academic Press, New York. 12. Pawlowskz S. (1982). Epidemiology and prevention of Taenia saginata infec- tion. In

  9. A purified inactivated Japanese encephalitis virus vaccine made in Vero cells.

    Science.gov (United States)

    Srivastava, A K; Putnak, J R; Lee, S H; Hong, S P; Moon, S B; Barvir, D A; Zhao, B; Olson, R A; Kim, S O; Yoo, W D; Towle, A C; Vaughn, D W; Innis, B L; Eckels, K H

    2001-08-14

    A second generation, purified, inactivated vaccine (PIV) against Japanese encephalitis (JE) virus was produced and tested in mice where it was found to be highly immunogenic and protective. The JE-PIV was made from an attenuated strain of JE virus propagated in certified Vero cells, purified, and inactivated with formalin. Its manufacture followed current GMP guidelines for the production of biologicals. The manufacturing process was efficient in generating a high yield of virus, essentially free of contaminating host cell proteins and nucleic acids. The PIV was formulated with aluminum hydroxide and administered to mice by subcutaneous inoculation. Vaccinated animals developed high-titered JE virus neutralizing antibodies in a dose dependent fashion after two injections. The vaccine protected mice against morbidity and mortality after challenge with live, virulent, JE virus. Compared with the existing licensed mouse brain-derived vaccine, JE-Vax, the Vero cell-derived JE-PIV was more immunogenic and as effective as preventing encephalitis in mice. The JE-PIV is currently being tested for safety and immunogenicity in volunteers.

  10. Preparation of Silicon by Calcium Reduction of Purified Rice Husk Ash

    International Nuclear Information System (INIS)

    Swe Zin Tun

    2011-12-01

    This research has studied on the possibility of production and preparation of silicon powder from rice husk ash (RHA) as raw material. RHA from gasifier, a waste product of the rice mill is rich in silica which contains 90.43% of silica. RHAs were purified by precipitation method using 1.5N, 2N, 2.5N and 3N of sodium hydroxide solution and 4.5N, 5N, 5.5N and 6.5N of sulphuric acid solution. The highest yield percent of silica was given by using 2.5N sodium hydroxide solution and 5N sulphuric acid solution X-ray fluoresence (XRF), X-rays diffraction (HRD) and Fourier transform infrared (FTRI) spectra were applied for determination of mineral content and chemical bonding in extracted silica and rice husk ash. Metallothermic reduction of purified extracted silica with calcium was investigated within the temperacture range of 700-900 C. The reduction product was characterized by XRD, XRF and scanning electron microcopy (SEM). The effect of temperature and reaction time in which reduction process was studied in this research.

  11. Biochemical Properties and Mechanism of Action of Enterocin LD3 Purified from Enterococcus hirae LD3.

    Science.gov (United States)

    Gupta, Aabha; Tiwari, Santosh Kumar; Netrebov, Victoria; Chikindas, Michael L

    2016-09-01

    Enterocin LD3 was purified using activity-guided multistep chromatography techniques such as cation-exchange and gel-filtration chromatography. The preparation's purity was tested using reverse-phase ultra-performance liquid chromatography. The specific activity was tested to be 187.5 AU µg(-1) with 13-fold purification. Purified enterocin LD3 was heat stable up to 121 °C (at 15 psi pressure) and pH 2-6. The activity was lost in the presence of papain, reduced by proteinase K, pepsin and trypsin, but was unaffected by amylase and lipase, suggesting proteinaceous nature of the compound and no role of carbohydrate and lipid moieties in the activity. MALDI-TOF/MS analysis of purified enterocin LD3 resolved m/z 4114.6, and N-terminal amino acid sequence was found to be H2NQGGQANQ-COOH suggesting a new bacteriocin. Dissipation of membrane potential, loss of internal ATP and bactericidal effect were recorded when indicator strain Micrococcus luteus was treated with enterocin LD3. It inhibited Gram-positive and Gram-negative bacteria including human pathogens such as Staphylococcus aureus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Salmonella typhi, Shigella flexneri, Listeria monocytogenes, Escherichia coli O157:H7, E. coli (urogenic, a clinical isolate) and Vibrio sp. These properties of purified enterocin LD3 suggest its applications as a food biopreservative and as an alternative to clinical antibiotics.

  12. Polymorphism in the Mr 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes

    International Nuclear Information System (INIS)

    Saboori, A.M.; Smith, B.L.; Agre, P.

    1988-01-01

    A M r 32,000 integral membrane protein has previously been identified on erythrocytes bearing the Rh(D) antigen and is thought to contain the antigenic variations responsible for the different Rh phenotypes. To study it on a biochemical level, a simple large-scale method was developed to purify the M r 32,000 Rh protein from multiple units of Rh(D)-positive and -negative blood. Erythrocyte membrane vesicles were solubilized in NaDodSO 4 , and a tracer of immunoprecipitated 125 I surface-labeled Rh protein was added. The Rh protein was purified to homogeneity by hydroxylapatite chromatography followed by preparative NaDodSO 4 /PAGE. Approximately 25 nmol of pure Rh protein was recovered from each unit of Rh(D)-positive and -negative blood. Rh protein purified from both Rh phenotypes appeared similar by one-dimensional NaDodSO 4 /PAGE, and the N-terminal amino acid sequences for the first 20 residues were identical. Rh proteins purified from Rh(D)-positive and -negative blood were compared by two-dimensional iodopeptide mapping after 125 I-labeling and α-chymotrypsin digestion. The peptide maps were very similar. These data indicate that a similar core Rh protein exists in both Rh(D)-positive and -negative erythrocytes, and the Rh proteins from erythrocytes with different Rh phenotypes contain distinct structural polymorphisms

  13. Population Level Purifying Selection and Gene Expression Shape Subgenome Evolution in Maize.

    Science.gov (United States)

    Pophaly, Saurabh D; Tellier, Aurélien

    2015-12-01

    The maize ancestor experienced a recent whole-genome duplication (WGD) followed by gene erosion which generated two subgenomes, the dominant subgenome (maize1) experiencing fewer deletions than maize2. We take advantage of available extensive polymorphism and gene expression data in maize to study purifying selection and gene expression divergence between WGD retained paralog pairs. We first report a strong correlation in nucleotide diversity between duplicate pairs, except for upstream regions. We then show that maize1 genes are under stronger purifying selection than maize2. WGD retained genes have higher gene dosage and biased Gene Ontologies consistent with previous studies. The relative gene expression of paralogs across tissues demonstrates that 98% of duplicate pairs have either subfunctionalized in a tissuewise manner or have diverged consistently in their expression thereby preventing functional complementation. Tissuewise subfunctionalization seems to be a hallmark of transcription factors, whereas consistent repression occurs for macromolecular complexes. We show that dominant gene expression is a strong determinant of the strength of purifying selection, explaining the inferred stronger negative selection on maize1 genes. We propose a novel expression-based classification of duplicates which is more robust to explain observed polymorphism patterns than the subgenome location. Finally, upstream regions of repressed genes exhibit an enrichment in transposable elements which indicates a possible mechanism for expression divergence. © The Author 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  14. Characterization of purified Sindbis virus nsP4 RNA-dependent RNA polymerase activity in vitro

    International Nuclear Information System (INIS)

    Rubach, Jon K.; Wasik, Brian R.; Rupp, Jonathan C.; Kuhn, Richard J.; Hardy, Richard W.; Smith, Janet L.

    2009-01-01

    The Sindbis virus RNA-dependent RNA polymerase (nsP4) is responsible for the replication of the viral RNA genome. In infected cells, nsP4 is localized in a replication complex along with the other viral non-structural proteins. nsP4 has been difficult to homogenously purify from infected cells due to its interactions with the other replication proteins and the fact that its N-terminal residue, a tyrosine, causes the protein to be rapidly turned over in cells. We report the successful expression and purification of Sindbis nsP4 in a bacterial system, in which nsP4 is expressed as an N-terminal SUMO fusion protein. After purification the SUMO tag is removed, resulting in the isolation of full-length nsP4 possessing the authentic N-terminal tyrosine. This purified enzyme is able to produce minus-strand RNA de novo from plus-strand templates, as well as terminally add adenosine residues to the 3' end of an RNA substrate. In the presence of the partially processed viral replicase polyprotein, P123, purified nsP4 is able to synthesize discrete template length minus-strand RNA products. Mutations in the 3' CSE or poly(A) tail of viral template RNA prevent RNA synthesis by the replicase complex containing purified nsP4, consistent with previously reported template requirements for minus-strand RNA synthesis. Optimal reaction conditions were determined by investigating the effects of time, pH, and the concentrations of nsP4, P123 and magnesium on the synthesis of RNA

  15. Epidemiology and genetic diversity of Taenia asiatica: a systematic review.

    Science.gov (United States)

    Ale, Anita; Victor, Bjorn; Praet, Nicolas; Gabriël, Sarah; Speybroeck, Niko; Dorny, Pierre; Devleesschauwer, Brecht

    2014-01-22

    Taenia asiatica has made a remarkable journey through the scientific literature of the past 50 years, starting with the paradoxical observation of high prevalences of T. saginata-like tapeworms in non-beef consuming populations, to the full description of its mitochondrial genome. Experimental studies conducted in the 1980s and 1990s have made it clear that the life cycle of T. asiatica is comparable to that of T. saginata, except for pigs being the preferential intermediate host and liver the preferential location of the cysts. Whether or not T. asiatica can cause human cysticercosis, as is the case for Taenia solium, remains unclear. Given the specific conditions needed to complete its life cycle, in particular the consumption of raw or poorly cooked pig liver, the transmission of T. asiatica shows an important ethno-geographical association. So far, T. asiatica has been identified in Taiwan, South Korea, Indonesia, the Philippines, Thailand, south-central China, Vietnam, Japan and Nepal. Especially this last observation indicates that its distribution is not restricted to South-East-Asia, as was thought so far. Indeed, the molecular tools developed over the last 20 years have made it increasingly possible to differentiate T. asiatica from other taeniids. Such tools also indicated that T. asiatica is related more closely to T. saginata than to T. solium, feeding the debate on its taxonomic status as a separate species versus a subspecies of T. saginata. Furthermore, the genetic diversity within T. asiatica appears to be very minimal, indicating that this parasite may be on the verge of extinction. However, recent studies have identified potential hybrids between T. asiatica and T. saginata, reopening the debate on the genetic diversity of T. asiatica and its status as a separate species.

  16. Epidemiology and genetic diversity of Taenia asiatica: a systematic review

    Science.gov (United States)

    2014-01-01

    Taenia asiatica has made a remarkable journey through the scientific literature of the past 50 years, starting with the paradoxical observation of high prevalences of T. saginata-like tapeworms in non-beef consuming populations, to the full description of its mitochondrial genome. Experimental studies conducted in the 1980s and 1990s have made it clear that the life cycle of T. asiatica is comparable to that of T. saginata, except for pigs being the preferential intermediate host and liver the preferential location of the cysts. Whether or not T. asiatica can cause human cysticercosis, as is the case for Taenia solium, remains unclear. Given the specific conditions needed to complete its life cycle, in particular the consumption of raw or poorly cooked pig liver, the transmission of T. asiatica shows an important ethno-geographical association. So far, T. asiatica has been identified in Taiwan, South Korea, Indonesia, the Philippines, Thailand, south-central China, Vietnam, Japan and Nepal. Especially this last observation indicates that its distribution is not restricted to South-East-Asia, as was thought so far. Indeed, the molecular tools developed over the last 20 years have made it increasingly possible to differentiate T. asiatica from other taeniids. Such tools also indicated that T. asiatica is related more closely to T. saginata than to T. solium, feeding the debate on its taxonomic status as a separate species versus a subspecies of T. saginata. Furthermore, the genetic diversity within T. asiatica appears to be very minimal, indicating that this parasite may be on the verge of extinction. However, recent studies have identified potential hybrids between T. asiatica and T. saginata, reopening the debate on the genetic diversity of T. asiatica and its status as a separate species. PMID:24450957

  17. The surface activity of purified ocular mucin at the air-liquid interface and interactions with meibomian lipids.

    Science.gov (United States)

    Millar, Thomas J; Tragoulias, Sophia T; Anderton, Philip J; Ball, Malcolm S; Miano, Fausto; Dennis, Gary R; Mudgil, Poonam

    2006-01-01

    Ocular mucins are thought to contribute to the stability of the tear film by reducing surface tension. The purpose of this study was to compare the effect of different mucins and hyaluronic acid (HA) alone and mixed with meibomian lipids on the surface pressure at an air-liquid interface. A Langmuir trough and Wilhelmy balance were used to measure and compare the surface activity of bovine submaxillary gland mucin (BSM), purified BSM, purified bovine ocular mucin and HA, and mixtures of these with meibomian lipids, phosphatidylcholine, and phosphatidylglycerol. Their appearance at the surface of an air-buffer interface was examined using epifluorescence microscopy. Purified ocular mucin had no surface activity even at concentrations that were 100 times more than normally occur in tears. By contrast, commercial BSM caused changes to surface pressure that were concentration dependent. The surface pressure-area profiles showed surface activity with maximum surface pressures of 12.3-22.5 mN/m depending on the concentration. Purified BSM showed no surface activity at low concentrations, whereas higher concentrations reached a maximum surface pressure of 25 mN/m. HA showed no surface activity, at low or high concentrations. Epifluorescence showed that the mucins were located at the air-buffer interface and changed the appearance of lipid films. Purified bovine ocular mucin and HA have no surface activity. However, despite having no surface activity in their own right, ocular mucins are likely to be present at the surface of the tear film, where they cause an increase in surface pressure by causing a compression of the lipids (a reorganization of the lipids) and alter the viscoelastic properties at the surface.

  18. Influence of lysozyme complexation with purified Aldrich humic acid on lysozyme activity

    NARCIS (Netherlands)

    Li, Y.; Tan, W.F.; Wang, M.X.; Liu, F.; Weng, L.P.; Norde, W.; Koopal, L.K.

    2012-01-01

    Humic acid is an important component of dissolved organic matter and in two previous papers it has been shown that purified Aldrich humic acid (PAHA) forms strong complexes with the oppositely charged protein lysozyme (LSZ). The complexation and aggregation of enzymes with humic acids may lead to

  19. Characterization of purified bacterial cellulose focused on its use on paper restoration.

    Science.gov (United States)

    Santos, Sara M; Carbajo, José M; Quintana, Ester; Ibarra, David; Gomez, Nuria; Ladero, Miguel; Eugenio, M Eugenia; Villar, Juan C

    2015-02-13

    Bacterial cellulose (BC) synthesized by Gluconacetobacter sucrofermentans CECT 7291 seems to be a good option for the restoration of degraded paper. In this work BC layers are cultivated and purified by two different methods: an alkaline treatment when the culture media contains ethanol and a thermal treatment if the media is free from ethanol. The main goal of these tests was the characterization of BC layers measured in terms of tear and burst indexes, optical properties, SEM, X-ray diffraction, FTIR, degree of polymerization, static and dynamic contact angles, and mercury intrusion porosimetry. The BC layers were also evaluated in the same terms after an aging treatment. Results showed that BC has got high crystallinity index, low internal porosity, good mechanical properties and high stability over time, especially when purified by the alkaline treatment. These features make BC an adequate candidate for degraded paper reinforcement. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Kinetic Analysis of Lactose Exchange in Proteoliposomes Reconstituted with Purified lac Permease

    NARCIS (Netherlands)

    Lolkema, Julius S.; Carrasco, Nancy; Kaback, H. Ronald

    1991-01-01

    Lactose exchange catalyzed by purified lac permease reconstituted into proteoliposomes was analyzed with unequal concentrations of lactose on either side of the membrane and at low pH so as to prevent equilibration of the two pools. Exchange with external concentrations below 1.0 mM is a

  1. Effect of purified gambir leaves extract to prevent atherosclerosis in rats

    Directory of Open Access Journals (Sweden)

    Nanang Yunarto

    2016-03-01

    , antiaterosklerosis AbstractBackground: Atherosclerosis is a risk factor for coronary heart disease (CHD. Catechin have highantioxidant activity that can prevent atherosclerosis. Gambir (Uncaria gambir, Roxb. leaves extract havehigh catechin content thereby potentially inhibiting atherosclerosis. This research was aimed to examineeffect of purified gambir leaves extract to prevent atherosclerosis in rats.Methods: The experimental laboratory study was conducted in Pharmacy Laboratory and Animal Laboratory,National Institute of Health Research and Development, Ministry of Health, Republic of Indonesia in 2014.Gambir leaves extract were purified to gain optimum catechin. Afterwards, antioxidant activity was testedusing 2.2-diphenyl-1-picrylhydrazyl (DPPH method, with ascorbic acid as positive control. Thirty six whitemale Sprague Dawley rats aged 2.5 months were randomly divided into six groups, i.e. normal control group,negative control group (aquadest, positive control group (atorvastatin 2 mg/200 g bw,extract dose I (20mg/200 g bw, dose II (40 mg/200 g bw and dose III (80 mg/200 g bw. The rats were given high fat diet andtreatment according to their group for 60 days, except for normal control group.Results: Catechin content in the purified gambir leaves extract was 92,69%. From antioxidant activity test, IC50 wasfound to be 11,76 μg/mL. Anti-atherosclerotic activity study shown that compared to negative control, all three dosesof purified gambir leaves extract were able to prevent atherosclerosis through inhibition of aortic wall thickening andfoam cell formation due to high fat diet (p<0.05. Anti-atherosclerotic activity increased with increasing dose.Conclusion: Gambir leaves purified extract had the effect of preventing the thickening of the walls andfoam cell formation rat aorta. (Health Science Journal of Indonesia 2015;6:105-10Keywords: gambir, catechin, antiatherosclerosis

  2. Echinococcus-Host Interactions at Cellular and Molecular Levels.

    Science.gov (United States)

    Brehm, K; Koziol, U

    2017-01-01

    The potentially lethal zoonotic diseases alveolar and cystic echinococcosis are caused by the metacestode larval stages of the tapeworms Echinococcus multilocularis and Echinococcus granulosus, respectively. In both cases, metacestode growth and proliferation occurs within the inner organs of mammalian hosts, which is associated with complex molecular host-parasite interactions that regulate nutrient uptake by the parasite as well as metacestode persistence and development. Using in vitro cultivation systems for parasite larvae, and informed by recently released, comprehensive genome and transcriptome data for both parasites, these molecular host-parasite interactions have been subject to significant research during recent years. In this review, we discuss progress in this field, with emphasis on parasite development and proliferation. We review host-parasite interaction mechanisms that occur early during an infection, when the invading oncosphere stage undergoes a metamorphosis towards the metacestode, and outline the decisive role of parasite stem cells during this process. We also discuss special features of metacestode morphology, and how this parasite stage takes up nutrients from the host, utilizing newly evolved or expanded gene families. We comprehensively review mechanisms of host-parasite cross-communication via evolutionarily conserved signalling systems and how the parasite signalling systems might be exploited for the development of novel chemotherapeutics. Finally, we point to an urgent need for the development of functional genomic techniques in this parasite, which will be imperative for hypothesis-driven analyses into Echinococcus stem cell biology, developmental mechanisms and immunomodulatory activities, which are all highly relevant for the development of anti-infective measures. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Hydrodynamic and Membrane Binding Properties of Purified Rous Sarcoma Virus Gag Protein

    Energy Technology Data Exchange (ETDEWEB)

    Dick, Robert A.; Datta, Siddhartha A.K.; Nanda, Hirsh; Fang, Xianyang; Wen, Yi; Barros, Marilia; Wang, Yun-Xing; Rein, Alan; Vogt, Volker M. (NCI); (Cornell); (CM); (NIST)

    2016-05-06

    Previously, no retroviral Gag protein has been highly purified in milligram quantities and in a biologically relevant and active form. We have purified Rous sarcoma virus (RSV) Gag protein and in parallel several truncation mutants of Gag and have studied their biophysical properties and membrane interactionsin vitro. RSV Gag is unusual in that it is not naturally myristoylated. From its ability to assemble into virus-like particlesin vitro, we infer that RSV Gag is biologically active. By size exclusion chromatography and small-angle X-ray scattering, Gag in solution appears extended and flexible, in contrast to previous reports on unmyristoylated HIV-1 Gag, which is compact. However, by neutron reflectometry measurements of RSV Gag bound to a supported bilayer, the protein appears to adopt a more compact, folded-over conformation. At physiological ionic strength, purified Gag binds strongly to liposomes containing acidic lipids. This interaction is stimulated by physiological levels of phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P2] and by cholesterol. However, unlike HIV-1 Gag, RSV Gag shows no sensitivity to acyl chain saturation. In contrast with full-length RSV Gag, the purified MA domain of Gag binds to liposomes only weakly. Similarly, both an N-terminally truncated version of Gag that is missing the MA domain and a C-terminally truncated version that is missing the NC domain bind only weakly. These results imply that NC contributes to membrane interactionin vitro, either by directly contacting acidic lipids or by promoting Gag multimerization.

    Retroviruses like HIV assemble at and bud from the plasma membrane of cells. Assembly requires the interaction between thousands of Gag molecules to form a lattice. Previous work indicated that lattice formation at the plasma membrane is influenced by the conformation of monomeric HIV. We have extended this work to the more tractable RSV Gag. Our

  4. Epidemiology and genetic diversity of Taenia asiatica: a systematic review.

    OpenAIRE

    Ale, Anita; Victor, Bjorn; Praet, Nicolas; Gabriël, Sarah; Speybroeck, Niko; Dorny, Pierre; Devleesschauwer, Brecht

    2014-01-01

    Taenia asiatica has made a remarkable journey through the scientific literature of the past 50 years, starting with the paradoxical observation of high prevalences of T. saginata-like tapeworms in non-beef consuming populations, to the full description of its mitochondrial genome. Experimental studies conducted in the 1980s and 1990s have made it clear that the life cycle of T. asiatica is comparable to that of T. saginata, except for pigs being the preferential intermediate host and liver th...

  5. Kinetics and Thermal Properties of Crude and Purified β-Galactosidase with Potential for the Production of Galactooligosaccharides

    Directory of Open Access Journals (Sweden)

    Anna Rafaela Cavalcante Braga

    2013-01-01

    Full Text Available β-Galactosidase is an enzyme that catalyzes the hydrolysis of lactose. It has potential importance due to various applications in the food and dairy industries, involving lactose-reduced ingredients. The properties of two β-galactosidase enzymes, crude and purified, from different sources, Kluyveromyces marxianus CCT 7082 and Kluyveromyces marxianus ATCC 16045, were analyzed. The pH and temperature optima, deactivation energy, thermal stability and kinetic and thermodynamic parameters were determined, as well as the ability to hydrolyze lactose and produce galactooligosaccharides. Purification process improved the properties of the enzymes, and the results showed that purified enzymes from both strains had a higher optimum temperature, and lower values of Km, thus showing greater affinity for o-nitrophenyl-β-D-galactopiranoside than the crude enzymes. The production of galactooligosaccharides was also greater when using purified enzymes, increasing the synthesis by more than 30 % by both strains.

  6. Comparison of solubilized and purified plasma membrane and nuclear insulin receptors

    International Nuclear Information System (INIS)

    Wong, K.Y.; Hawley, D.; Vigneri, R.; Goldfine, I.D.

    1988-01-01

    Prior studies have detected biochemical and immunological differences between insulin receptors in plasma membranes and isolated nuclei. To further investigate these receptors, they were solubilized in Triton X-100 partially purified by wheat germ agglutinin-agarose chromatography. In these preparations, the nuclear and plasma membrane receptors had very similar pH optima (pH 8.0) and reactivities to a group of polyclonal antireceptor antibodies. Further, both membrane preparations had identical binding activities when labeled insulin was competed for by unlabeled insulin (50% inhibition at 800 pM). Next, nuclear and plasma membranes were solubilized and purified to homogeneity by wheat germ agglutinin-agarose and insulin-agarose chromatography. In both receptors, labeled insulin was covalently cross-linked to a protein of 130 kilodaltons representing the insulin receptor α subunit. When preparations of both receptors were incubated with insulin and then adenosine 5'-[γ- 32 P]triphosphate, a protein of 95 kilodaltons representing the insulin receptor β subunit was phosphorylated in a dose-dependent manner. These studies indicate, therefore, that solubilized plasma membrane and nuclear insulin receptors have similar structures and biochemical properties, and they suggest that they are the same (or very similar) proteins

  7. Cloning and characterization of the fatty acid-binding protein gene from the protoscolex of Taenia multiceps.

    Science.gov (United States)

    Nie, Hua-Ming; Xie, Yue; Fu, Yan; Yang, Ying-Dong; Gu, Xiao-Bin; Wang, Shu-Xian; Peng, Xi; Lai, Wei-Ming; Peng, Xue-Rong; Yang, Guang-You

    2013-05-01

    Taenia multiceps (Cestoda: Taeniidae), a worldwide cestode parasite, is emerging as an important helminthic zoonosis due to serious or fatal central nervous system disease commonly known as coenurosis in domestic and wild ruminants including humans. Herein, a fatty acid-binding protein (FABP) gene was identified from transcriptomic data in T. multiceps. This gene, which contains a complete coding sequence, was amplified by reverse-transcriptase polymerase chain reaction. The corresponding protein, which was named TmFABP, had a molecular weight of 14 kDa, and subsequently was recombinantly expressed in Escherichia coli. The fusion protein was purified on Ni-NTA beads (Bio-Rad). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analyses showed that the purified recombinant protein caused immunogenicity. Immunohistochemical studies showed that TmFABP was expressed at the tegumental level in the protoscolices and in the cells between the body wall and parenchyma layer of the cestode. In sections from gravid proglottids, intense staining was detected in the uterus and eggs. Based on this, TmFABP could be switched on during differentiation of germinative layers to protoscoleces and from metacestodes to adult worms. Taken together, our results already reported for T. multiceps suggest the possibility of TmFABP developing a vaccine to control and prevent coenurosis.

  8. Sympatric Distribution of Three Human Taenia Tapeworms Collected between 1935 and 2005 in Korea

    Science.gov (United States)

    Jeon, Hyeong-Kyu; Kim, Kyu-Heon; Chai, Jong-Yil; Yang, Hyun-Jong; Rim, Han-Jong

    2008-01-01

    Taeniasis has been known as one of the prevalent parasitic infections in Korea. Until recently, Taenia saginata had long been considered a dominant, and widely distributed species but epidemiological profiles of human Taenia species in Korea still remain unclear. In order to better understand distribution patterns of human Taenia tapeworms in Korea, partial nucleotide sequences of mitochondrial cox1 and ITS2 (internal transcribed spacer 2) were determined, along with morphological examinations, on 68 Taenia specimens obtained from university museum collections deposited since 1935. Genomic DNA was extracted from formalin-preserved specimens. Phylogenetic relationships among the genotypes (cox1 haplotype) detected in this study were inferred using the neighbor-joining method as a tree building method. Morphological and genetic analyses identified 3 specimens as T. solium, 51 specimens as T. asiatica, and 14 specimens as T. saginata. Our results indicate that all 3 Taenia tapeworms are sympatrically distributed in Korea with T. asiatica dominating over T. saginata and T. solium. PMID:19127329

  9. C@Fe 3 O 4 /NTA-Ni magnetic nanospheres purify histidine-tagged ...

    African Journals Online (AJOL)

    This study reports synthesis of Ni-nitrilotriacetic acid (Ni-NTA) modified carbon nanospheres containing magnetic Fe3O4 particles (C@Fe3O4), which can act as a general tool to separate and purify histidine-tagged fetidin. In this experiment, C nanospheres are prepared from glucose using the hydrothermal process, ...

  10. On the preparation of as-produced and purified single-walled carbon nanotube samples for standardized X-ray diffraction characterization

    International Nuclear Information System (INIS)

    Allaf, Rula M.; Rivero, Iris V.; Spearman, Shayla S.; Hope-Weeks, Louisa J.

    2011-01-01

    The aim of this research was to specify proper sample conditioning for acquiring representative X-ray diffraction (XRD) profiles for single-walled carbon nanotube (SWCNT) samples. In doing so, a specimen preparation method for quantitative XRD characterization of as-produced and purified arc-discharge SWCNT samples has been identified. Series of powder XRD profiles were collected at different temperatures, states, and points of time to establish appropriate conditions for acquiring XRD profiles without inducing much change to the specimen. It was concluded that heating in the 300-450 deg. C range for 20 minutes, preferably vacuum-assisted, and then sealing the sample is an appropriate XRD specimen preparation technique for purified arc-discharge SWCNT samples, while raw samples do not require preconditioning for characterization. - Graphical Abstract: A sample preparation method for XRD characterization of as-produced and purified arc-discharge SWCNT samples is identified. The preparation technique seeks to acquire representative XRD profiles without inducing changes to the samples. Purified samples required 20 minutes of heating at (300-450)deg. C, while raw samples did not require preconditioning for characterization. Highlights: → Purification routines may induce adsorption onto the SWCNT samples. → Heating a SWCNT sample may result in material loss, desorption, and SWCNTs closing. → Raw arc-discharge samples do not require preparation for XRD characterization. → Heating is appropriate specimen preparation for purified and heat-treated samples. → XRD data fitting is required for structural analysis of SWCNT bundles.

  11. Ferromagnetic Levan Composite: An Affinity Matrix to Purify Lectin

    Directory of Open Access Journals (Sweden)

    Renata Angeli

    2009-01-01

    Full Text Available A simple and inexpensive procedure used magnetite and levan to synthesize a composite recovered by a magnetic field. Lectins from Canavalia ensiformis (Con A and Cratylia mollis (Cramoll 1 and Cramoll 1,4 did bind specifically to composite. The magnetic property of derivative favored washing out contaminating proteins and recovery of pure lectins with glucose elution. Cramoll 1 was purified by this affinity binding procedure in two steps instead of a previous three-step protocol with ammonium sulfate fractionation, affinity chromatography on Sephadex G-75, and ion exchange chromatography through a CM-cellulose column.

  12. Immobilized Metal Affinity Chromatography Co-Purifies TGF-β1 with Histidine-Tagged Recombinant Extracellular Proteins

    Science.gov (United States)

    Kaur, Jasvir; Reinhardt, Dieter P.

    2012-01-01

    Extracellular recombinant proteins are commonly produced using HEK293 cells as histidine-tagged proteins facilitating purification by immobilized metal affinity chromatography (IMAC). Based on gel analyses, this one-step purification typically produces proteins of high purity. Here, we analyzed the presence of TGF-β1 in such IMAC purifications using recombinant extracellular fibrillin-1 fragments as examples. Analysis of various purified recombinant fibrillin-1 fragments by ELISA consistently revealed the presence of picomolar concentrations of active and latent TGF-β1, but not of BMP-2. These quantities of TGF-β1 were not detectable by Western blotting and mass spectrometry. However, the amounts of TGF-β1 were sufficient to consistently trigger Smad2 phosphorylation in fibroblasts. The purification mechanism was analyzed to determine whether the presence of TGF-β1 in these protein preparations represents a specific or non-specific co-purification of TGF-β1 with fibrillin-1 fragments. Control purifications using conditioned medium from non-transfected 293 cells yielded similar amounts of TGF-β1 after IMAC. IMAC of purified TGF-β1 and the latency associated peptide showed that these proteins bound to the immobilized nickel ions. These data clearly demonstrate that TGF-β1 was co-purified by specific interactions with nickel, and not by specific interactions with fibrillin-1 fragments. Among various chromatographic methods tested for their ability to eliminate TGF-β1 from fibrillin-1 preparations, gel filtration under high salt conditions was highly effective. As various recombinant extracellular proteins purified in this fashion are frequently used for experiments that can be influenced by the presence of TGF-β1, these findings have far-reaching implications for the required chromatographic schemes and quality controls. PMID:23119075

  13. Characterization of Affinity-Purified Isoforms of Acinetobacter calcoaceticus Y1 Glutathione Transferases

    Directory of Open Access Journals (Sweden)

    Chin-Soon Chee

    2014-01-01

    Full Text Available Glutathione transferases (GST were purified from locally isolated bacteria, Acinetobacter calcoaceticus Y1, by glutathione-affinity chromatography and anion exchange, and their substrate specificities were investigated. SDS-polyacrylamide gel electrophoresis revealed that the purified GST resolved into a single band with a molecular weight (MW of 23 kDa. 2-dimensional (2-D gel electrophoresis showed the presence of two isoforms, GST1 (pI 4.5 and GST2 (pI 6.2 with identical MW. GST1 was reactive towards ethacrynic acid, hydrogen peroxide, 1-chloro-2,4-dinitrobenzene, and trans,trans-hepta-2,4-dienal while GST2 was active towards all substrates except hydrogen peroxide. This demonstrated that GST1 possessed peroxidase activity which was absent in GST2. This study also showed that only GST2 was able to conjugate GSH to isoproturon, a herbicide. GST1 and GST2 were suggested to be similar to F0KLY9 (putative glutathione S-transferase and F0KKB0 (glutathione S-transferase III of Acinetobacter calcoaceticus strain PHEA-2, respectively.

  14. Characterization of Affinity-Purified Isoforms of Acinetobacter calcoaceticus Y1 Glutathione Transferases

    Science.gov (United States)

    Chee, Chin-Soon; Tan, Irene Kit-Ping; Alias, Zazali

    2014-01-01

    Glutathione transferases (GST) were purified from locally isolated bacteria, Acinetobacter calcoaceticus Y1, by glutathione-affinity chromatography and anion exchange, and their substrate specificities were investigated. SDS-polyacrylamide gel electrophoresis revealed that the purified GST resolved into a single band with a molecular weight (MW) of 23 kDa. 2-dimensional (2-D) gel electrophoresis showed the presence of two isoforms, GST1 (pI 4.5) and GST2 (pI 6.2) with identical MW. GST1 was reactive towards ethacrynic acid, hydrogen peroxide, 1-chloro-2,4-dinitrobenzene, and trans,trans-hepta-2,4-dienal while GST2 was active towards all substrates except hydrogen peroxide. This demonstrated that GST1 possessed peroxidase activity which was absent in GST2. This study also showed that only GST2 was able to conjugate GSH to isoproturon, a herbicide. GST1 and GST2 were suggested to be similar to F0KLY9 (putative glutathione S-transferase) and F0KKB0 (glutathione S-transferase III) of Acinetobacter calcoaceticus strain PHEA-2, respectively. PMID:24892084

  15. Oxidation of aromatic alcohols by purified methanol dehydrogenase from Methylosinus trichosporium.

    OpenAIRE

    Mountfort, D O

    1990-01-01

    Methanol dehydrogenase was found to be present in subcellular preparations of methanol-grown Methylosinus trichosporium and occurred almost wholly in the soluble fraction of the cell. The enzyme, purified by DEAE-Sephadex and Sephadex G-100 chromatography, showed broad specificity toward different substrates and oxidized the aromatic alcohols benzyl, vanillyl, and veratryl alcohols in addition to a range of aliphatic primary alcohols. No enzyme activity was found toward the corresponding alde...

  16. 75 FR 6211 - Prospective Grant of Exclusive License: Purified Inactivated Dengue Tetravalent Vaccine...

    Science.gov (United States)

    2010-02-08

    ... Exclusive License: Purified Inactivated Dengue Tetravalent Vaccine Containing a Common 30 Nucleotide Deletion in the 3'-UTR of Dengue Types 1,2,3, and 4 AGENCY: National Institutes of Health, Public Health...., ``Development of Mutations Useful for Attenuating Dengue Viruses and Chimeric Dengue Viruses''-- European Patent...

  17. Dry fumes purifying system using anhydrous baking soda; Procede chimique d`epuration des fumees au bicarbonate de soude anhydre

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    1998-04-01

    UNISYSTEMS has developed the industrial implementation of the chemical process using anhydrous backing soda, patented by SOLVAY, for purifying fumes containing inorganic salts and sulphur oxides as polluting agents. The system can be applied to industrial processes releasing this type of polluting agents in the fumes at a temperature over 160 deg C, as it is specially indicated in purifying fumes coming from ceramic firing kilns. (authors)

  18. Taeniasis and Cysticercosis as A Zoonotic Parasitic Disease

    Directory of Open Access Journals (Sweden)

    Sarwitri Endah Estuningsih

    2009-06-01

    Full Text Available Taeniasis is a parasitic disease caused by tapeworms from the genus Taenia, and infection with the larvae form of Taenia is called Cysticercosis. Some species of Taenia are zoonotic, and humans serve as the definitive host, the intermediate host or both. Humans are the definitive hosts for Taenia solium, T. saginata and T. asiatica, however, humans also act as an intermediate host for T. solium and T. asiatica. Animals, such as pigs, are the intermediate host for T. solium and T. asiatica, and cattle are the intermediate host for T. saginata. Humans can be infected by taeniasis when they eat beef or pork that contains larvae (cysticercus. While, cysticercosis is transmitted via food or water contaminated with the eggs of Taenia spp. The transmission may also occur by autoinfection due to lack of hygiene. The diagnosis of taeniasis based on finding the eggs or proglotid in the human feces. For diagnosing cysticercosis in live animals can be done by tongue palpation to find the presence of cysts or nodules. Serological test may also help for diagnosing cysticercosis in humans or animals. Adult tapeworms in the intestine can be killed by anthelmintic and prevention of taeniasis can be conducted by avoiding raw or undercooked pork (T. solium and T. asiatica and beef (T. saginata. Besides that, to prevent the infection of T. solium, T. saginata or T. asiatica, pigs or cattle should not be exposed to human feces.

  19. Magnetism for understanding catalyst analysis of purified carbon nanotubes

    Energy Technology Data Exchange (ETDEWEB)

    Bellouard, Christine; Mercier, Guillaume; Cahen, Sébastien; Ghanbaja, Jaafar; Medjahdi, Ghouti [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France); Gleize, Jérôme [Laboratoire de Chimie Physique-Approche Multi-échelle de Milieux Complexes-Université de Lorraine, 1 Bd Arago, 57078 Metz (France); Lamura, Gianrico [CNR-SPIN – Dipartimento di Fisica, via Dodecaneso 33, 16146 Genova (Italy); Hérold, Claire [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France); Vigolo, Brigitte, E-mail: Brigitte.Vigolo@univ-lorraine.fr [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France)

    2016-08-01

    The precise quantification of catalyst residues in purified carbon nanotubes is often a major issue in view of any fundamental and/or applicative studies. More importantly, since the best CNTs are successfully grown with magnetic catalysts, their quantification becomes strictly necessary to better understand intrinsic properties of CNT. For these reasons, we have deeply analyzed the catalyst content remained in nickel–yttrium arc-discharge single walled carbon nanotubes purified by both a chlorine-gas phase and a standard acid-based treatment. The study focuses on Ni analysis which has been investigated by transmission electron microscopy, X-ray diffraction, thermogravimetry analysis, and magnetic measurements. In the case of the acid-based treatment, all quantifications result in a decrease of the nanocrystallized Ni by a factor of two. In the case of the halogen gas treatment, analysis and quantification of Ni content is less straightforward: a huge difference appears between X-ray diffraction and thermogravimetry results. Thanks to magnetic measurements, this disagreement is explained by the presence of Ni{sup 2+} ions, belonging to NiCl{sub 2} formed during the Cl-based purification process. In particular, NiCl{sub 2} compound appears under different magnetic/crystalline phases: paramagnetic or diamagnetic, or well intercalated in between carbon sheets with an ordered magnetic phase at low temperature. - Highlights: • Cl-gas treatment of Ni catalyst of carbon nanotubes leads to NiCl{sub 2} residue. • Magnetic measurements show the transformation of Ni{sup 0} in Ni{sup 2+}through a purification process. • High temperature Cl treatment removes 75% of metallic impurities. • Cl-purification yields to an amount of metal of 1.5% in arc-discharge CNT samples.

  20. CHARACTERIZATION OF THE PARTIALLY PURIFIED PLANTARCIN SR18 PRODUCED BY LACTOBACILLUS PLANTARUM SR18

    Directory of Open Access Journals (Sweden)

    Wagih El-Shouny

    2013-04-01

    Full Text Available The bacteriocin bound to the cells and that secreted into the culture filtrate of Lactobacillus plantarum SR18 were precipitated by 75% ammomium sulphate, dialysed and further purified by Gel filtration on Sephadex G-100. Bacteriocins were purified from proteins bound to the cell of L. plantarum SR18 (plantarcin SR18 a and culture filtrate proteins (plantarcin SR18 b, respectively. The SDS-PAGE of partially purified Plantarcin SR18a showed a molecular weight of 3.5 KDa. While, plantarcin SR18 b had a molecular weight of 10.3 KDa. The antibacterial activity of the tested plantarcin SR18 preparations suffered no measurable loss after 45 min at 80ºC. Whereas, At 100ºC, significant decrease in the activity of bacteriocin preparations (60- 80 % took place by the end of 45 min. At pH ranged from 5-8, the activity of the plantarcin SR18 preparations suffered no measurable loss. Dissociating agents significantly affected the bacteriocin activity. Thus, tween 80 and mercaptoethanol increased the activity of bacteriocin preparations to 1.2-1.4 fold. Sodium dodecyl sulphate (SDS increased the activity of the tested bacteriocin preparations by about 20%.The lowest residual activity (60% was recorded after treatment with Triton X100 for 45 min. Protease completely inhibited the activities of all forms of plantarcin SR18 after 45 min at 37ºC.

  1. 78 FR 78812 - Purified Carboxymethylcellulose From the Netherlands: Final Results of Antidumping Duty...

    Science.gov (United States)

    2013-12-27

    ... Carboxymethylcellulose From the Netherlands: Final Results of Antidumping Duty Administrative Review and Final No... Netherlands. For the final results, we continue to find that sales of subject merchandise by Akzo Nobel... of the AD order on purified CMC from the Netherlands.\\1\\ We invited interested parties to comment on...

  2. Influence of a highly purified senna extract on colonic epithelium.

    Science.gov (United States)

    van Gorkom, B A; Karrenbeld, A; van Der Sluis, T; Koudstaal, J; de Vries, E G; Kleibeuker, J H

    2000-01-01

    Chronic use of sennoside laxatives often causes pseudomelanosis coli. A recent study suggested that pseudomelanosis coli is associated with an increased colorectal cancer risk. A single high dose of highly purified senna extract increased proliferation rate and reduced crypt length in the sigmoid colon compared to historical controls. To evaluate in a controlled study the effects of highly purified senna extract on cell proliferation and crypt length in the entire colon and on p53 and bcl-2 expression. Addition of a senna extract to colonic lavage was studied in 184 consecutive outpatients. From 32 randomised patients, 15 with sennosides (Sen), 17 without (NSen), biopsies were taken. Proliferative activity was studied in 4 areas of the colon, using 5-bromo-2'-deoxyuridine labelling and immunohistochemistry (labelling index, LI). Expression of p53 and bcl-2 in the sigmoid colon was determined immunohistochemically. Crypts were shorter in Sen than in NSen in the transverse and sigmoid colon. LI was higher in Sen than in NSen in the entire colon. No difference in p53 expression was seen. Bcl-2 expression was higher in both groups when crypts were shorter and/or proliferation was increased. Sennosides induce acute massive cell loss probably by apoptosis, causing shorter crypts, and increased cell proliferation and inhibition of apoptosis to restore cellularity. These effects may reflect the mechanism for the suggested cancer-promoting effect of chronic sennoside use. Copyright 2000 S. Karger AG, Basel

  3. Surface plasmon resonance sensing: from purified biomolecules to intact cells.

    Science.gov (United States)

    Su, Yu-Wen; Wang, Wei

    2018-04-12

    Surface plasmon resonance (SPR) has become a well-recognized label-free technique for measuring the binding kinetics between biomolecules since the invention of the first SPR-based immunosensor in 1980s. The most popular and traditional format for SPR analysis is to monitor the real-time optical signals when a solution containing ligand molecules is flowing over a sensor substrate functionalized with purified receptor molecules. In recent years, rapid development of several kinds of SPR imaging techniques have allowed for mapping the dynamic distribution of local mass density within single living cells with high spatial and temporal resolutions and reliable sensitivity. Such capability immediately enabled one to investigate the interaction between important biomolecules and intact cells in a label-free, quantitative, and single cell manner, leading to an exciting new trend of cell-based SPR bioanalysis. In this Trend Article, we first describe the principle and technical features of two types of SPR imaging techniques based on prism and objective, respectively. Then we survey the intact cell-based applications in both fundamental cell biology and drug discovery. We conclude the article with comments and perspectives on the future developments. Graphical abstract Recent developments in surface plasmon resonance (SPR) imaging techniques allow for label-free mapping the mass-distribution within single living cells, leading to great expansions in biomolecular interactions studies from homogeneous substrates functionalized with purified biomolecules to heterogeneous substrates containing individual living cells.

  4. Photovoltaic performance of TiO2 electrode adsorbed with gardenia yellow purified by nonionic polymeric sorbent in dye-sensitized solar cells.

    Science.gov (United States)

    Kwon, Oh Oun; Kim, Eui Jin; Lee, Jae Hyeok; Kim, Tae Young; Park, Kyung Hee; Kim, Sang Yook; Suh, Hwa Jin; Lee, Hyo Jung; Lee, Jae Wook

    2015-02-05

    To improve the photovoltaic conversion efficiency in dye-sensitized solar cells (DSSCs), TiO2 electrode adsorbed with gardenia yellow purified by nonionic polymeric sorbent was successfully formulated on nanoporous TiO2 surface. Adsorption and desorption properties of crude gardenia yellow solution on a macroporous resin, XAD-1600, were investigated to purify gardenia yellow because of its strong adsorption and desorption abilities as well as high selectivity. To this end, adsorption equilibrium and kinetic data were measured and fitted using adsorption isotherms and kinetic models. Adsorption and desorption breakthrough curves in a column packed with XAD-1600 resin was obtained to optimize the separation process of gardenia yellow. The photovoltaic performance of the photo-electrode adsorbed with the crude and purified gardenia yellow in DSSCs was compared from current-voltage measurements. The results showed that the photovoltaic conversion efficiency was highly dependent on how to separate and purify gardenia yellow as a photosensitizer. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Phospholipid environment alters hormone-sensitivity of the purified insulin receptor kinase.

    OpenAIRE

    Lewis, R E; Czech, M P

    1987-01-01

    Insulin receptor kinase, affinity-purified by adsorption and elution from immobilized insulin, is stimulated 2-3-fold by insulin in detergent solution. Reconstitution of the receptor kinase into leaky vesicles containing phosphatidylcholine and phosphatidylethanolamine (1:1, w/w) by detergent removal on Sephadex G-50 results in the complete loss of receptor kinase sensitivity to activation by insulin. Insulin receptors in these vesicles also exhibit an increase in their apparent affinity for ...

  6. Tuberculin purified protein derivative-reactive T cells in cord blood lymphocytes.

    OpenAIRE

    Shiratsuchi, H; Tsuyuguchi, I

    1981-01-01

    Lymphocytes obtained from cord blood of newborn babies who were born of healthy mothers were studied in vitro for their responsiveness to purified protein derivative (PPD) of tuberculin. Cord blood lymphocytes proliferated in vitro by stimulation with PPD, despite wide variations in the results. Studies with fractionated lymphocytes revealed that PPD-responding cells belonged to E-rosetting, nylon wool-nonadherent T lymphocytes. Non-E-rosetting B lymphocytes alone did not proliferate at all a...

  7. The effect of purified sewage discharge from a sewage treatment plant on the physicochemical state of water in the receiver

    Directory of Open Access Journals (Sweden)

    Kanownik Włodzimierz

    2016-09-01

    Full Text Available The paper presents changes in the contents of physicochemical indices of the Sudół stream water caused by a discharge of purified municipal sewage from a small mechanical-biological treatment plant with throughput of 300 m3·d−1 and a population equivalent (p.e. – 1,250 people. The discharge of purified sewage caused a worsening of the stream water quality. Most of the studied indices values increased in water below the treatment plant. Almost a 100-fold increase in ammonium nitrogen, 17-fold increase in phosphate concentrations and 12-fold raise in BOD5 concentrations were registered. Due to high values of these indices, the water physicochemical state was below good. Statistical analysis revealed a considerable effect of the purified sewage discharge on the stream water physicochemical state. A statistically significant increase in 10 indices values (BOD5, COD-Mn, EC, TDS, Cl−, Na+, K+, PO43−, N-NH4+ and N-NO2 as well as significant decline in the degree of water saturation with oxygen were noted below the sewage treatment plant. On the other hand, no statistically significant differences between the water indices values were registered between the measurement points localised 150 and 1,000 m below the purified sewage discharge. It evidences a slow process of the stream water self-purification caused by an excessive loading with pollutants originating from the purified sewage discharge.

  8. Evaluation of monoclonal anti-A and anti-B and affinity-purified Ulex europaeus lectin I for forensic blood grouping.

    Science.gov (United States)

    Gaensslen, R E; Lee, H C; Carroll, J E

    1984-01-01

    Two different monoclonal anti-A and anti-B and several different affinity purified Ulex europaeus lectin I reagents were evaluated and compared with conventional anti-A and anti-B sera and Ulex anti-H for serologic properties, in inhibition tests with secretor salivas, and in elution tests with bloodstains. The monoclonal and purified reagents were found to be comparable to conventional ones, and accordingly suitable for forensic inhibition and elution procedures.

  9. Experimental studies on removal of airborne haloanisoles by non-thermal plasma air purifiers

    DEFF Research Database (Denmark)

    Fang, Lei; Hallam, David; Bermúdez, Raúl

    2016-01-01

    A laboratory study was conducted to test the performance of non-thermal plasma air purifiers on its removal effectiveness of two haloanisoles – 2,4,6-trichloroanisole (TCA) and 2,4,6-Tribromoanisole (TBA). TCA and TBA are the two major compounds found in wine cellars that can contaminate wine to ...

  10. Gamma ray irradiation to semi-purified diet

    International Nuclear Information System (INIS)

    Takigawa, Akihiro; Danbara, Hiroshi; Ohyama, Yoshinobu

    1976-01-01

    Semi-purified diet containing 10% soybean oil was irradiated with gamma rays at levels of 0.6, 3 and 6 Mrad and was fed to chicks. Crude fat contents of the diets decreased and a considerable amount of peroxide was formed with high doses of irradiation. Feed consumption and feed efficiency of the highly irradiated diets were less than those of control. Metabolizable energy and digestibility of the diets, especially of fat, were decreased with the irradiation. The chicks fed with irradiated diets showed marked dilatation of the small intestine and the liver, and their erythrocytes were more fragile than those of control. The same phenomena were found with the chicks fed the diet containing the oil highly oxidized by autoxidation. Irradiation of the diet excluding oil showed little effect on the growth of chicks. It was considered that these phenomena were caused by the peroxide or other oxidation products of fat which were formed with gamma ray irradiation. (auth.)

  11. Subpopulations in purified platelets adhering on glass.

    Science.gov (United States)

    Donati, Alessia; Gupta, Swati; Reviakine, Ilya

    2016-06-22

    Understanding how platelet activation is regulated is important in the context of cardiovascular disorders and their management with antiplatelet therapy. Recent evidence points to different platelet subpopulations performing different functions. In particular, procoagulant and aggregating subpopulations have been reported in the literature in platelets treated with the GPVI agonists. How the formation of platelet subpopulations upon activation is regulated remains unclear. Here, it is shown that procoagulant and aggregating platelet subpopulations arise spontaneously upon adhesion of purified platelets on clean glass surfaces. Calcium ionophore treatment of the adhering platelets resulted in one platelet population expressing both the procoagulant and the adherent population markers phosphatidylserine and the activated form of GPIIb/IIIa, while all of the platelets expressed CD62P independently of the ionophore treatment. Therefore, all platelets have the capacity to express all three activation markers. It is concluded that platelet subpopulations observed in various studies reflect the dynamics of the platelet activation process.

  12. A long-term aging study of honeycomb drift tubes for the HERA-B Outer Tracker using a circulated and purified CF$_{4}$ gas mixture

    CERN Document Server

    Capéans-Garrido, M; Hohlmann, M; Schmidt, B

    2003-01-01

    The Outer Tracker of HERA-B uses a gas mixture containing CF/sub 4/ to obtain high electron drift velocities. The high cost of this gas makes it necessary to circulate the gas mixture which must then be purified to avoid accumulation of air and pollutants. However, the usage of gas purifiers poses the danger of outgassing pollutants from the purifiers themselves into the gas stream. Purifiers could also be attacked chemically by the aggressive products from the cracking of CF/sub 4/ molecules in the plasma avalanches of the detector. This could potentially release further harmful pollutants into the gas stream. To test for such effects, a long-term irradiation study of about 3000 h was carried out with the honeycomb drift tubes that are used in the Outer Tracker. This provided a check of the long-term stability of the gas purifiers before putting them into operation for the full-size detector. We report on the experimental setup, procedures and the results obtained. (8 refs).

  13. Clinical studies of a purified timothy pollen extract: desensitization therapy with a purified timothy pollen preparation compared to a crude timothy pollen extract. II. Results of the tests in vitro and their relation to symptoms and tests in vivo.

    Science.gov (United States)

    Nordvall, S L; Berg, T; Johansson, S G; Lanner, A

    1982-01-01

    Perennial desensitization therapy was given during a period of 3.5 years to 40 children allergic to grass pollen allergens. 20 patients were treated with a crude and another 20 with a purified timothy pollen extract. 8 children served as untreated controls. The concentration of total and specific IgE in the treated groups covaried with those in the control group. Neither a suppression of the seasonal booster effect nor a suppression of IgE synthesis attributable to the treatment was found. The rise of timothy-specific "blocking' IgG antibodies was more pronounced in the group treated with the purified extract than in the group treated with the crude extract. A significant difference was found only after 3.5 years of treatment. The amplitude of rise of IgG antibodies correlated significantly with the effect of the treatment as judged by repeated conjunctival titration test. The results suggest that a good IgG response is an indication of successful therapy and that a better IgG response may be achieved with purified allergen extracts.

  14. Functional reconstitution into liposomes of purified human RhCG ammonia channel.

    Directory of Open Access Journals (Sweden)

    Isabelle Mouro-Chanteloup

    Full Text Available BACKGROUND: Rh glycoproteins (RhAG, RhBG, RhCG are members of the Amt/Mep/Rh family which facilitate movement of ammonium across plasma membranes. Changes in ammonium transport activity following expression of Rh glycoproteins have been described in different heterologous systems such as yeasts, oocytes and eukaryotic cell lines. However, in these complex systems, a potential contribution of endogenous proteins to this function cannot be excluded. To demonstrate that Rh glycoproteins by themselves transport NH(3, human RhCG was purified to homogeneity and reconstituted into liposomes, giving new insights into its channel functional properties. METHODOLOGY/PRINCIPAL FINDINGS: An HA-tag introduced in the second extracellular loop of RhCG was used to purify to homogeneity the HA-tagged RhCG glycoprotein from detergent-solubilized recombinant HEK293E cells. Electron microscopy analysis of negatively stained purified RhCG-HA revealed, after image processing, homogeneous particles of 9 nm diameter with a trimeric protein structure. Reconstitution was performed with sphingomyelin, phosphatidylcholine and phosphatidic acid lipids in the presence of the C(12E(8 detergent which was subsequently removed by Biobeads. Control of protein incorporation was carried out by freeze-fracture electron microscopy. Particle density in liposomes was a function of the Lipid/Protein ratio. When compared to empty liposomes, ammonium permeability was increased two and three fold in RhCG-proteoliposomes, depending on the Lipid/Protein ratio (1/300 and 1/150, respectively. This strong NH(3 transport was reversibly inhibited by mercuric and copper salts and exhibited a low Arrhenius activation energy. CONCLUSIONS/SIGNIFICANCE: This study allowed the determination of ammonia permeability per RhCG monomer, showing that the apparent Punit(NH3 (around 1x10(-3 microm(3xs(-1 is close to the permeability measured in HEK293E cells expressing a recombinant human RhCG (1.60x10

  15. Preparation of sol-gel TiO2/purified Na-bentonite composites and their photovoltaic application for natural dye-sensitized solar cells

    International Nuclear Information System (INIS)

    Saelim, Ni-on; Magaraphan, Rathanawan; Sreethawong, Thammanoon

    2011-01-01

    Highlights: → Natural dye from red cabbage was successfully employed in DSSC. → A fast sol-gel method to produce TiO 2 /clay thin film was proposed. → The sol-gel-prepared TiO 2 /clay was applied as the scattering layer on top of TiO 2 electrode. → Thicker sol-gel-prepared TiO 2 /clay electrode showed higher DSSC efficiency. - Abstract: The sol-gel TiO 2 /purified natural clay electrodes having Ti:Si molar ratios of 95:5 and 90:10 were initially prepared, sensitized with natural red cabbage dye, and compared to the sol-gel TiO 2 electrode in terms of physicochemical characteristics and solar cell efficiency. The results showed that the increase in purified Na-bentonite content greatly increased the specific surface area and total pore volume of the prepared sol-gel TiO 2 /purified Na-bentonite composites because the clay platelets prevented TiO 2 particle agglomeration. The sol-gel TiO 2 /5 mol% Si purified Na-bentonite and sol-gel TiO 2 /10 mol% Si purified Na-bentonite composites could increase the film thickness of solar cells without cracking when they were coated as a scattering layer on the TiO 2 semiconductor-based film, leading to increasing the efficiency of the natural dye-sensitized solar cells in this work.

  16. Process for the winning of a concentrate containing uranium and purified phosphoric acid, as well as the concentrate containing uranium and purified phosphoric acid obtained by this process

    International Nuclear Information System (INIS)

    1980-01-01

    The uranium containing concentrate and purified phosphoric acid are obtained by treating wet phosphoric acid with an inorganic fluorine compound (ammonium fluoride) and an aliphatic ketone (acetone) in the presence of a reducing agent (finely divided iron). The ketone is added first and the formed uranium precipitate is separated from the solution. If the fluorine compound is added first, the yield is lowered by a factor of 2. (Th.P.)

  17. Proteomic characterisation of bovine and avian purified protein derivatives and identification of specific antigens for serodiagnosis of bovine tuberculosis

    OpenAIRE

    Infantes-Lorenzo, José Antonio; Moreno, Inmaculada; Risalde, María de los Ángeles; Roy, Álvaro; Villar, Margarita; Romero, Beatriz; Ibarrola, Nieves; de la Fuente, José; Puentes, Eugenia; de Juan, Lucía; Gortázar, Christian; Bezos, Javier; Domínguez, Lucas; Domínguez, Mercedes

    2017-01-01

    Background Bovine purified protein derivative (bPPD) and avian purified protein derivative (aPPD) are widely used for bovine tuberculosis diagnosis. However, little is known about their qualitative and quantitative characteristics, which makes their standardisation difficult. In addition, bPPD can give false-positive tuberculosis results because of sequence homology between Mycobacterium bovis (M. bovis) and M. avium proteins. Thus, the objective of this study was to carry out a proteomic cha...

  18. Towards the international collaboration for detection, surveillance and control of taeniasis/ cysticercosis and echinococcosis in Asia and the Pacific.

    Science.gov (United States)

    Ito, Akira; Wandra, Toni; Sato, Marcello O; Mamuti, Wulamu; Xiao, Ning; Sako, Yasuhito; Nakao, Minoru; Yamasaki, Hiroshi; Nakaya, Kazuhiro; Okamoto, Munehiro; Craig, Philip S

    2006-01-01

    Both cysticercosis and echinococcosis are potentially among the most serious helminth zoonoses threatening human health worldwide. However, due to the lack of reliable tools for confirmation or identification of patients or infected animals, epidemiological data are expected to be underestimated. Conversely, sometimes, such data are over estimated due to the lack of specificity. The most important issue for doing field surveys is that they use evidence based science. In this communication, advanced immunological and molecular tools for detection of individuals infected with either metacestodes or adult tapeworms are briefly overviewed, and the applications of such tools for epidemiological surveys in Indonesia, China and other countries are introduced. As immunological tools are based on antigen-antibody responses, there may exist some cross-reactions. Therefore, immunodiagnostic tools are expected to be useful for primary screening, and should be combined with confirmation of direct parasitological evidence (morphology or DNA), and imaging techniques for cysts. As a risk factor for human cysticercosis is the presence of tapeworm carriers, detection of taeniasis cases and differentiation of the three human Taenia species (Taenia solium, T. saginata and T. asiatica) in Asia and the Pacific requires consideration. Similarly, in northwest China, Echinococcus granulosus and E. multilocularis are coendemic and differentiation of these species is required in humans and definitive hosts. It is stressed that combination of several tools for identification of the parasite and for confirmation of diseases is important for obtaining highly reliable data before consideration of control of these zoonoses. Recent projects coordinated by Asahikawa Medical College have concentrated on immunological and molecular diagnostic techniques transferable to colleagues from endemic regions of Asia and the Pacific, and on organization of two international symposia to establish a platform

  19. 75 FR 61700 - Purified Carboxymethylcellulose From Finland, the Netherlands, and Sweden: Final Results of the...

    Science.gov (United States)

    2010-10-06

    ... also referred to as purified sodium CMC, polyanionic cellulose, or cellulose gum, which is a white to....gov/frn . The paper copy and electronic version of the Decision Memo are identical in content. Final...

  20. In vitro and in vivo inhibitory effects of some fungicides on catalase produced and purified from white-rot fungus Phanerochaete chrysosporium.

    Science.gov (United States)

    Kavakçıoğlu, Berna; Tarhan, Leman

    2014-10-01

    In this study, in vitro and in vivo effects of some commonly used fungicides, antibiotics, and various chemicals on isolated and purified catalase from Phanerochaete chrysosporium were investigated. The catalase was purified 129.10-fold by using 60% ammonium sulfate and 60% ethanol precipitations, DEAE-cellulose anion exchange and Sephacryl-S-200 gel filtration chromatographies from P. chrysosporium growth in carbon- and nitrogen-limited medium for 12 days. The molecular weight of native purified catalase from P. chrysosporium was found to be 290 ± 10 kDa, and sodium dodecyl sulfate (SDS)-PAGE results indicated that enzyme consisted of four apparently identical subunits, with a molecular weight of 72.5 ± 2.5 kDa. Kinetic characterization studies showed that optimum pH and temperature, Km and Vmax values of the purified catalase which were stable in basic region and at comparatively high temperatures were 7.5, 30°C, 289.86 mM, and 250,000 U/mg, respectively. The activity of purified catalase from P. chrysosporium was significantly inhibited by dithiothreitol (DTT), 2-mercaptoethanol, iodoacetamide, EDTA, and sodium dodecyl sulfate (SDS). It was found that while antibiotics had no inhibitory effects, 45 ppm benomyl, 144 ppm captan, and 47.5 ppm chlorothalonil caused 14.52, 10.82, and 38.86% inhibition of purified catalase, respectively. The inhibition types of these three fungicides were found to be non-competitive inhibition with the Ki values of 1.158, 0.638, and 0.145 mM and IC50 values of 0.573, 0.158, 0.010 mM, respectively. The results of in vivo experiments also showed that benomyl, captan and chlorothalonil caused 15.25, 1.96, and 36.70% activity decreases after 24-h treatments compared to that of the control.

  1. Taeniasis and cysticercosis in Bali and North Sumatra, Indonesia.

    Science.gov (United States)

    Wandra, Toni; Depary, A A; Sutisna, Putu; Margono, Sri S; Suroso, Thomas; Okamoto, Munehiro; Craig, Philip S; Ito, Akira

    2006-01-01

    It has been reported that three human Taenia species are distributed in Indonesia: Taenia solium, Taenia asiatica and Taenia saginata. T. asiatica is well known in North Sumatra, especially on Samosir island in Lake Toba. T. solium and T. saginata are known from Bali. T. solium is most serious public health issue in Papua (former Irian Jaya). In this report, we briefly review the present situation of these three human Taenia species mainly in Bali and North Sumatra. For community based epidemiological survey, we have adopted and applied questionnaire, microscopic observation of eggs, coproantigen tests, coproDNA tests, mitochondrial DNA analysis and morphology for isolated specimens, serology for cysticercosis and taeniasis. For detection of taeniasis cases of both T. saginata and T. asiatica, questionnaire by expert doctors is perfect to detect worm carriers. Corpoantigen test developed for detection of taeniasis of T. solium is also highly reliable to detect taeniasis carriers in the community. Epidemiological data from these two islands 2002-2005 are reviewed with backgrounds of historical culture, religions and customs.

  2. New host, geographical records, and factors affecting the prevalence of helminths infection from synanthropic rodents in Yucatán, Mexico

    OpenAIRE

    Panti-May J. A.; Palomo-Arjona E.; Gurubel-González Y.; Torres-Castro M. A.; Vidal-Martínez V. M.; Machain-Williams C.; Hernández-Betancourt S. F.; Del Rosario Robles M.

    2017-01-01

    The aim of this paper was to study the occurrence of helminths in Mus musculus and Rattus rattus from urban, suburban and rural settlements in Yucatán, Mexico; and to analyse the host factors (e.g. sex) related to helminths’ distribution. Helminths in a total of 279 rodents were surveyed by visual examination of the liver for metacestodes and faecal examination for helminth eggs using the formalin-ethyl acetate sedimentation technique. The cestodes Hydatigera taeniaeformis (metacestodes detec...

  3. A study of the process of two staged anaerobic fermentation as a possible method for purifying sewage

    Energy Technology Data Exchange (ETDEWEB)

    Inoue, Y; Kouama, K; Matsuo, T

    1983-01-01

    Great attention has recently been given to the study of the processes of anaerobic fermentation, which may become alternatives to the existing methods for purifying waste waters which use aerobic microorganisms. A series of experimentswere conducted with the use of an artificially prepared liquid (fermented milk and starch) which imitates the waste to be purified, in order to explain the capabilities of the process of two staged anaerobic fermentation (DAS) as a method for purifying waste waters. The industrial system of the process includes: a fermentation vat for acetic fermentation with recirculation of the sediment, a primary settler, a fermentation tank for methane fermentation and a secondary settler. The process was conducted at a loading speed (based on Carbon) from 0.15 to 0.4 kilograms per cubic meter per day at a temperature of 38C. The degree of conversion of the fermented organic substances into volatile organic acids was not a function of the loading speed and was 54 to 57 percent in the acetic fermentation tank, where 95 to 97 percent of the organic material was broken down with the production of methane and carbon dioxide.

  4. Application of aqueous biphasic systems as strategy to purify tannase from Aspergillus tamarii URM 7115.

    Science.gov (United States)

    de Sena, Amanda Reges; Barros Oliveira, Flávio Manoel; Campos Leite, Tonny Cley; Evaristo da Silva Nascimento, Talita Camila; Moreira, Keila Aparecida; de Assis, Sandra Aparecida

    2017-10-21

    The aims of the current study are to assess the influence of polyethylene glycol (PEG) concentration, molar mass, pH, and citrate concentrations on aqueous biphasic systems based on 2 4 factorial designs, as well as to check their capacity to purify tannase secreted by Aspergillus tamarii URM 7115. Tannase was produced through submerged fermentation at 26°C for 67 h in Czapeck-Dox modified broth and added with yeast extract and tannic acid. The factorial design was followed to assess the influence of PEG molar mass (M PEG 600; 4,000 and 8,000 g/ mol), and PEG (C PEG 20.0; 22.0 and 24.0% w/w) and citrate concentrations (C CIT 15.0, 17.5, and 20.0%, w/w), as well as of pH (6.0, 7.0, and 8.0) on the response variables; moreover, partition coefficient (K), yield (Y), and purification factor (PF) were analyzed. The most suitable parameters to purify tannase secreted by A. tamarii URM 7115 through a biphasic system were 600 (g/mol) M PEG , 24% (w/w) C PEG , 15% (w/w) C CIT at pH 6.0 and they resulted in 6.33 enzyme partition, 131.25% yield, 19.80 purification factor and 195.08 selectivity. Tannase secreted by A. tamarii URM 7115 purified through aqueous biphasic systems composed of PEG/citrate can be used for industrial purposes, since it presents suitable purification factor and yield.

  5. Determining inhibition effects of some aromatic compounds on peroxidase enzyme purified from white and red cabbage

    Energy Technology Data Exchange (ETDEWEB)

    Öztekin, Aykut, E-mail: aoztekin@agri.edu.tr [Ataturk University, Science Faculty, Department of Chemistry, 25240-Erzurum (Turkey); Agri Ibrahim Cecen University Faculty of Arts and Sciences, Department of Chemistry, 04100-Agri (Turkey); Almaz, Züleyha, E-mail: zturkoglu-2344@hotmail.com [Ataturk University, Science Faculty, Department of Chemistry, 25240-Erzurum (Turkey); Mus Alparslan University Faculty of Sciences, Department of Moleculer Biology, 49250-Mus (Turkey); Özdemir, Hasan, E-mail: hozdemir@atauni.edu.tr [Ataturk University, Science Faculty, Department of Chemistry, 25240-Erzurum (Turkey)

    2016-04-18

    Peroxidases (E.C.1.11.1.7) catalyze the one electron oxidation of wide range of substrates. They are used in synthesis reaction, removal of peroxide from industrial wastes, clinical biochemistry and immunoassays. In this study, the white cabbage (Brassica Oleracea var. capitata f. alba) and red cabbage (Brassica oleracea L. var. capitata f. rubra) peroxidase enzymes were purified for investigation of inhibitory effect of some aromatic compounds on these enzymes. IC{sub 50} values and Ki constants were calculated for the molecules of 6-Amino nicotinic hydrazide, 6-Amino-5-bromo nicotinic hydrazide, 2-Amino-5-hydroxy benzohydrazide, 4-Amino-3-hydroxy benzohydrazide on purified enzymes and inhibition type of these molecules were determined. (This research was supported by Ataturk University. Project Number: BAP-2015/98).

  6. Determining inhibition effects of some aromatic compounds on peroxidase enzyme purified from white and red cabbage

    Science.gov (United States)

    Öztekin, Aykut; Almaz, Züleyha; Özdemir, Hasan

    2016-04-01

    Peroxidases (E.C.1.11.1.7) catalyze the one electron oxidation of wide range of substrates. They are used in synthesis reaction, removal of peroxide from industrial wastes, clinical biochemistry and immunoassays. In this study, the white cabbage (Brassica Oleracea var. capitata f. alba) and red cabbage (Brassica oleracea L. var. capitata f. rubra) peroxidase enzymes were purified for investigation of inhibitory effect of some aromatic compounds on these enzymes. IC50 values and Ki constants were calculated for the molecules of 6-Amino nicotinic hydrazide, 6-Amino-5-bromo nicotinic hydrazide, 2-Amino-5-hydroxy benzohydrazide, 4-Amino-3-hydroxy benzohydrazide on purified enzymes and inhibition type of these molecules were determined. (This research was supported by Ataturk University. Project Number: BAP-2015/98).

  7. Determining inhibition effects of some aromatic compounds on peroxidase enzyme purified from white and red cabbage

    International Nuclear Information System (INIS)

    Öztekin, Aykut; Almaz, Züleyha; Özdemir, Hasan

    2016-01-01

    Peroxidases (E.C.1.11.1.7) catalyze the one electron oxidation of wide range of substrates. They are used in synthesis reaction, removal of peroxide from industrial wastes, clinical biochemistry and immunoassays. In this study, the white cabbage (Brassica Oleracea var. capitata f. alba) and red cabbage (Brassica oleracea L. var. capitata f. rubra) peroxidase enzymes were purified for investigation of inhibitory effect of some aromatic compounds on these enzymes. IC_5_0 values and Ki constants were calculated for the molecules of 6-Amino nicotinic hydrazide, 6-Amino-5-bromo nicotinic hydrazide, 2-Amino-5-hydroxy benzohydrazide, 4-Amino-3-hydroxy benzohydrazide on purified enzymes and inhibition type of these molecules were determined. (This research was supported by Ataturk University. Project Number: BAP-2015/98).

  8. 75 FR 15678 - Certain Purified Carboxymethylcellulose from the Netherlands: Extension of Time Limit for...

    Science.gov (United States)

    2010-03-30

    ... Carboxymethylcellulose from the Netherlands: Extension of Time Limit for Preliminary Results of Antidumping Duty...) from the Netherlands. The period of review is July 1, 2008, through June 30, 2009. This extension is... of the antidumping duty order on purified CMC from the Netherlands. See Initiation of Antidumping and...

  9. Differentiation of purified malignant B cells induced by PMA or by activated normal T cells

    NARCIS (Netherlands)

    van Kooten, C.; Rensink, I.; Aarden, L.; van Oers, R.

    1993-01-01

    We studied the in vitro differentiation (immunoglobulin production) of purified malignant B cells of 21 patients with different B-cell malignancies, including chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), hairy cell leukemia (HCL) and non-Hodgkin lymphoma (NHL). Direct

  10. Evaluation of protection factors of a breath-responsive-powered air-purifying respirator

    International Nuclear Information System (INIS)

    Nakagawa, Masahiro; Nojima, Shun; Fujii, Katsutoshi; Shishido, Nobuhito; Sakai, Toshiya; Umehara, Takashi; Shimizu, Isamu

    2012-01-01

    It is essential to wear an air-purifying respirator in the radiation works in a contaminated atmosphere. A breath-responsive-powered air-purifying respirator (BR-PAPR) has been recently developed. However, no research has yet been conducted to determine the protection factor (PF) of the BR-PAPR in actual workplaces. In this study, the PFs of the BR-PAPR were measured by a man-test apparatus and compared with those of a non-powered full face mask. The PFs were measured under three different situations; normal wearing condition, clogging the filter and leaving a gap between the face and the mask. Under these situations, it was found that the PFs of the BR-PAPR are higher than those of the non-powered full face mask. PFs greater than 4,000 were obtained for 95% of the subjects who wear the BR-PAPR, and PFs over 6,667, the upper limit of the man-test apparatus, were obtained for 49% of them. The questionnaire survey was conducted for workers. The results showed that the workers feel a reduced burden when they wear the BR-PAPR. The results of this study showed high protection performance and operation efficiency of the BR-PAPR. (author)

  11. An Experiment with Air Purifiers in Delhi during Winter 2015-2016

    OpenAIRE

    Vyas, Sangita; Srivastav, Nikhil; Spears, Dean

    2016-01-01

    Particulate pollution has important consequences for human health, and is an issue of global concern. Outdoor air pollution has become a cause for alarm in India in particular because recent data suggest that ambient pollution levels in Indian cities are some of the highest in the world. We study the number of particles between 0.5μm and 2.5μm indoors while using affordable air purifiers in the highly polluted city of Delhi. Though substantial reductions in indoor number concentrations are ob...

  12. Infection of Taenia asiatica in a Bai Person in Dali, China.

    Science.gov (United States)

    Wang, Li; Luo, Xuenong; Hou, Junling; Guo, Aijiang; Zhang, Shaohua; Li, Hailong; Cai, Xuepeng

    2016-02-01

    We report here a human case of Taenia asiatica infection which was confirmed by genetic analyses in Dali, China. A patient was found to have symptoms of taeniasis with discharge of tapeworm proglottids. By sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene, we observed nucleotide sequence identity of 99% with T. asiatica and 96% with T. saginata. Using the cytochrome b (cytb) gene, 99% identity with T. asiatica and 96% identity with T. saginata were found. Our findings suggest that taeniasis of people in Dali, China may be mainly caused by T. asiatica.

  13. LH-RH binding to purified pituitary plasma membranes: absence of adenylate cyclase activation.

    Science.gov (United States)

    Clayton, R N; Shakespear, R A; Marshall, J C

    1978-06-01

    Purified bovine pituitary plasma membranes possess two specific LH-RH binding sites. The high affinity site (2.5 X 10(9) l/mol) has low capacity (9 X 10(-15) mol/mg membrane protein) while the low affinity site 6.1 X 10(5) l/mol) has a much higher capacity (1.1 X 10(-10) mol/mg). Specific LH-RH binding to plasma membranes is increased 8.5-fold during purification from homogenate whilst adenylate cyclase activity is enriched 7--8-fold. Distribution of specific LH-RH binding to sucrose density gradient interface fractions parallels that of adenylate cyclase activity. Mg2+ and Ca2+ inhibit specific [125I]LH-RH binding at micromolar concentrations. Synthetic LH-RH, up to 250 microgram/ml, failed to stimulate adenylase cyclase activity of the purified bovine membranes. Using a crude 10,800 g rat pituitary membrane preparation, LH-RH similarly failed to activate adenylate cyclase even in the presence of guanyl nucleotides. These data confirm the presence of LH-RH receptor sites on pituitary plasma membranes and suggest that LH-RH-induced gonadotrophin release may be mediated by mechanisms other than activation of adenylate cyclase.

  14. 77 FR 14733 - Purified Carboxymethylcellulose From Finland and the Netherlands: Extension of Time Limit for...

    Science.gov (United States)

    2012-03-13

    ... DEPARTMENT OF COMMERCE International Trade Administration [A-405-803, A-421-811] Purified Carboxymethylcellulose From Finland and the Netherlands: Extension of Time Limit for Preliminary Results of Antidumping... carboxymethylcellulose from Finland and the Netherlands covering the period July 1, 2010, through June 30, 2011. See...

  15. Application of decolourized and partially purified polygalacturonase and α-amylase in apple juice clarification

    Directory of Open Access Journals (Sweden)

    Tapati Bhanja Dey

    2014-01-01

    Full Text Available Polygalacturonase and α-amylase play vital role in fruit juice industry. In the present study, polygalacturonase was produced by Aspergillus awamori Nakazawa MTCC 6652 utilizing apple pomace and mosambi orange (Citrus sinensis var mosambi peels as solid substrate whereas, α-amylase was produced from A. oryzae (IFO-30103 using wheat bran by solid state fermentation (SSF process. These carbohydrases were decolourized and purified 8.6-fold, 34.8-fold and 3.5-fold, respectively by activated charcoal powder in a single step with 65.1%, 69.8% and 60% recoveries, respectively. Apple juice was clarified by these decolourized and partially purified enzymes. In presence of 1% polygalacturonase from mosambi peels (9.87 U/mL and 0.4% α-amylase (899 U/mL, maximum clarity (%T660nm = 97.0% of juice was attained after 2 h of incubation at 50 ºC in presence of 10 mM CaCl2. Total phenolic content of juice was reduced by 19.8% after clarification, yet with slightly higher %DPPH radical scavenging property.

  16. Application of decolourized and partially purified polygalacturonase and α-amylase in apple juice clarification.

    Science.gov (United States)

    Dey, Tapati Bhanja; Banerjee, Rintu

    2014-01-01

    Polygalacturonase and α-amylase play vital role in fruit juice industry. In the present study, polygalacturonase was produced by Aspergillus awamori Nakazawa MTCC 6652 utilizing apple pomace and mosambi orange (Citrus sinensis var mosambi) peels as solid substrate whereas, α-amylase was produced from A. oryzae (IFO-30103) using wheat bran by solid state fermentation (SSF) process. These carbohydrases were decolourized and purified 8.6-fold, 34.8-fold and 3.5-fold, respectively by activated charcoal powder in a single step with 65.1%, 69.8% and 60% recoveries, respectively. Apple juice was clarified by these decolourized and partially purified enzymes. In presence of 1% polygalacturonase from mosambi peels (9.87 U/mL) and 0.4% α-amylase (899 U/mL), maximum clarity (%T(660 nm) = 97.0%) of juice was attained after 2 h of incubation at 50 °C in presence of 10 mM CaCl2. Total phenolic content of juice was reduced by 19.8% after clarification, yet with slightly higher %DPPH radical scavenging property.

  17. Antigenic activity of concentrated purified cultural rabies vaciine inactivated by gamma-rays

    Energy Technology Data Exchange (ETDEWEB)

    Morogova, V M; Krutilina, D.V.; Latypova, R G; Dulina, A V; Nigamov, F N; Pogrebnyak, E M [Ufimskij Inst. Vaktsin i Syvorotok; Sanitarno-Ehpidemiologicheskaya Stantsiya, Ufimskaya Gorodskaya [USSR

    1978-12-01

    The ability to stimulate the production of virus-neutralizing antibodies and the reactivity of concentrated, purified and gamma-inactivated cultural antirabic vaccine from the Vnukovo-32 strain (35-38th passages) were studied in experiments with humans and animals. After two intramuscular immunizations (2 ml) at 21- or 23-day intervals, this preparation of the vaccine yielded antibody titers (both in humans and in animals) not lower than those obtained after a full course of immunization with the cultural or cerebral antirabic vaccine.

  18. Antioxidant and ACE Inhibitory Bioactive Peptides Purified from Egg Yolk Proteins

    Directory of Open Access Journals (Sweden)

    Marwa Yousr

    2015-12-01

    Full Text Available Protein by-products from the extraction of lecithin from egg yolk can be converted into value-added products, such as bioactive hydrolysates and peptides that have potential health enhancing antioxidant, and antihypertensive properties. In this study, the antioxidant and angiotensin converting enzyme (ACE inhibitory activities of peptides isolated and purified from egg yolk protein were investigated. Defatted egg yolk was hydrolyzed using pepsin and pancreatin and sequentially fractionated by ultrafiltration, followed by gel filtration to produce egg yolk gel filtration fractions (EYGF. Of these, two fractions, EYGF-23 and EYGF-33, effectively inhibited the peroxides and thiobarbituric acid reactive substance (TBARS in an oxidizing linoleic acid model system. The antioxidant mechanism involved superoxide anion and hydroxyl radicals scavenging and ferrous chelation. The presence of hydrophobic amino acids such as tyrosine (Y and tryptophan (W, in sequences identified by LC-MS as WYGPD (EYGF-23 and KLSDW (EYGF-33, contributed to the antioxidant activity and were not significantly different from the synthetic BHA antioxidant. A third fraction (EYGF-56 was also purified from egg yolk protein by gel filtration and exhibited high ACE inhibitory activity (69% and IC50 value (3.35 mg/mL. The SDNRNQGY peptide (10 mg/mL had ACE inhibitory activity, which was not significantly different from that of the positive control captopril (0.5 mg/mL. In addition, YPSPV in (EYGF-33 (10 mg/mL had higher ACE inhibitory activity compared with captopril. These findings indicated a substantial potential for producing valuable peptides with antioxidant and ACE inhibitory activity from egg yolk.

  19. Antioxidant and ACE Inhibitory Bioactive Peptides Purified from Egg Yolk Proteins.

    Science.gov (United States)

    Yousr, Marwa; Howell, Nazlin

    2015-12-07

    Protein by-products from the extraction of lecithin from egg yolk can be converted into value-added products, such as bioactive hydrolysates and peptides that have potential health enhancing antioxidant, and antihypertensive properties. In this study, the antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of peptides isolated and purified from egg yolk protein were investigated. Defatted egg yolk was hydrolyzed using pepsin and pancreatin and sequentially fractionated by ultrafiltration, followed by gel filtration to produce egg yolk gel filtration fractions (EYGF). Of these, two fractions, EYGF-23 and EYGF-33, effectively inhibited the peroxides and thiobarbituric acid reactive substance (TBARS) in an oxidizing linoleic acid model system. The antioxidant mechanism involved superoxide anion and hydroxyl radicals scavenging and ferrous chelation. The presence of hydrophobic amino acids such as tyrosine (Y) and tryptophan (W), in sequences identified by LC-MS as WYGPD (EYGF-23) and KLSDW (EYGF-33), contributed to the antioxidant activity and were not significantly different from the synthetic BHA antioxidant. A third fraction (EYGF-56) was also purified from egg yolk protein by gel filtration and exhibited high ACE inhibitory activity (69%) and IC50 value (3.35 mg/mL). The SDNRNQGY peptide (10 mg/mL) had ACE inhibitory activity, which was not significantly different from that of the positive control captopril (0.5 mg/mL). In addition, YPSPV in (EYGF-33) (10 mg/mL) had higher ACE inhibitory activity compared with captopril. These findings indicated a substantial potential for producing valuable peptides with antioxidant and ACE inhibitory activity from egg yolk.

  20. Phylogenetic systematics of the genus Echinococcus (Cestoda: Taeniidae).

    Science.gov (United States)

    Nakao, Minoru; Lavikainen, Antti; Yanagida, Tetsuya; Ito, Akira

    2013-11-01

    Echinococcosis is a serious helminthic zoonosis in humans, livestock and wildlife. The pathogenic organisms are members of the genus Echinococcus (Cestoda: Taeniidae). Life cycles of Echinococcus spp. are consistently dependent on predator-prey association between two obligate mammalian hosts. Carnivores (canids and felids) serve as definitive hosts for adult tapeworms and their herbivore prey (ungulates, rodents and lagomorphs) as intermediate hosts for metacestode larvae. Humans are involved as an accidental host for metacestode infections. The metacestodes develop in various internal organs, particularly in liver and lungs. Each metacestode of Echinococcus spp. has an organotropism and a characteristic form known as an unilocular (cystic), alveolar or polycystic hydatid. Recent molecular phylogenetic studies have demonstrated that the type species, Echinococcus granulosus, causing cystic echinococcosis is a cryptic species complex. Therefore, the orthodox taxonomy of Echinococcus established from morphological criteria has been revised from the standpoint of phylogenetic systematics. Nine valid species including newly resurrected taxa are recognised as a result of the revision. This review summarises the recent advances in the phylogenetic systematics of Echinococcus, together with the historical backgrounds and molecular epidemiological aspects of each species. A new phylogenetic tree inferred from the mitochondrial genomes of all valid Echinococcus spp. is also presented. The taxonomic nomenclature for Echinococcus oligarthrus is shown to be incorrect and this name should be replaced with Echinococcus oligarthra. Copyright © 2013 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  1. Rheological properties of purified illite clays in glycerol/water suspensions

    Science.gov (United States)

    Dusenkova, I.; Malers, J.; Berzina-Cimdina, L.

    2015-04-01

    There are many studies about rheological properties of clay-water suspensions, but no published investigations about clay-glycerol suspensions. In this work apparent viscosity of previously purified illite containing clay fraction clay minerals were almost totally removed by centrifugation. All obtained suspensions behaved as shear-thinning fluids with multiple times higher viscosity than pure glycerol/water solutions. Reduction of clay fraction concentration by 5% decreased the apparent viscosity of 50% glycerol/water suspensions approximately 5 times. There was basically no difference in apparent viscosity between all four 50% glycerol/water suspensions, but in 90% glycerol/water suspensions samples from Iecava deposit showed slightly higher apparent viscosity, which could be affected by the particle size distribution.

  2. Antifungal properties of wheat histones (H1-H4) and purified wheat histone H1

    Science.gov (United States)

    Wheat (Triticum sp.) histones H1, H2, H3, and H4 were extracted. H1 was further purified. Their activities against fungi with varying degrees of wheat pathogenicity were determined. They included Aspergillus flavus, A. fumigatus, A. niger, F. oxysporum, F. verticillioides, F. solani, F. graminearu...

  3. [Prokaryotic expression and histological localization of the Taenia solium CDC37 gene].

    Science.gov (United States)

    Huang, Jiang; Li, Bo; Dai, Jia-Lin; Zhang, Ai-Hua

    2013-02-01

    To express Taenia solium gene encoding cell division cycle 37 protein (TsCDC37) and investigate its antigenicity and localization in adults of Taenia solium. The complete coding sequence of TsCDC37 was amplified by PCR based on the recombinant plasmid clone from the cDNA library of adult Taenia solium. The PCR product was cloned into a prokaryotic expression vector pET-28a (+). The recombinant expression plasmid was identified by PCR, double endonuclease digestion and sequencing. The recombinant plasmid was transformed into E. coli BL21/DE3 and followed by expression of the protein induced by IPTG. The mice were immunized subcutaneously with purified recombinant TsCDC37 formulated in Freund's adjuvant. The antigenicity of the recombinant protein was examined by Western blotting. The localization of TsCDC37 in adult worms was demonstrated by immunofluorescent technique. The recombinant expression vector was constructed successfully. The recombinant protein was about M(r) 52 000, it was then purified and specifically recognized by immuno sera of SD rats and sera from patients infected with Taenia solium, Taenia saginata or Taenia asiatica. The immunofluorescence assay revealed that TsCDC37 located at the tegument of T. solium adult and the eggs. TsCDC37 gene has been expressed with immunoreactivity. The recombinant protein is mainly expressed in tegument and egg, and is a common antigen of the three human taenia cestodes.

  4. Removal efficiency of water purifier and adsorbent for iodine, cesium, strontium, barium and zirconium in drinking water.

    Science.gov (United States)

    Sato, Itaru; Kudo, Hiroaki; Tsuda, Shuji

    2011-01-01

    The severe incident of Fukushima Daiichi Nuclear Power Station has caused radioactive contamination of environment including drinking water. Radioactive iodine, cesium, strontium, barium and zirconium are hazardous fission products because of the high yield and/or relatively long half-life. In the present study, 4 pot-type water purifiers and several adsorbents were examined for the removal effects on these elements from drinking water. Iodide, iodate, cesium and barium were removed by all water purifiers with efficiencies about 85%, 40%, 75-90% and higher than 85%, respectively. These efficiencies lasted for 200 l, which is near the recommended limits for use of filter cartridges, without decay. Strontium was removed with initial efficiencies from 70% to 100%, but the efficiencies were slightly decreased by use. Zirconium was removed by two models, but hardly removed by the other models. Synthetic zeolite A4 efficiently removed cesium, strontium and barium, but had no effect on iodine and zirconium. Natural zeolite, mordenite, removed cesium with an efficiency as high as zeolite A4, but the removal efficiencies for strontium and barium were far less than those of zeolite A4. Activated carbon had little removal effects on these elements. In case of radioactive contamination of tap water, water purifiers may be available for convenient decontamination of drinking water in the home.

  5. The addition of medium-chain triglycerides to a purified fish oil-based diet alters inflammatory profiles in mice.

    Science.gov (United States)

    Carlson, Sarah J; Nandivada, Prathima; Chang, Melissa I; Mitchell, Paul D; O'Loughlin, Alison; Cowan, Eileen; Gura, Kathleen M; Nose, Vania; Bistrian, Bruce R; Puder, Mark

    2015-02-01

    Parenteral nutrition associated liver disease (PNALD) is a deadly complication of long term parenteral nutrition (PN) use in infants. Fish oil-based lipid emulsion has been shown in recent years to effectively treat PNALD. Alternative fat sources free of essential fatty acids have recently been investigated for health benefits related to decreased inflammatory response. We hypothesized that the addition of medium-chain triglycerides (MCT) to a purified fish oil-based diet would decrease the response to inflammatory challenge in mice, while allowing for sufficient growth and development. Six groups of ten adult male C57/Bl6 mice were pair-fed different dietary treatments for a period of twelve weeks, varying only in fat source (percent calories by weight): 10.84% soybean oil (SOY), 10% coconut oil (HCO), 10% medium-chain triglycerides (MCT), 3% purified fish oil (PFO), 3% purified fish oil with 3% medium-chain triglycerides (50:50 MCT:PFO) and 3% purified fish oil with 7.59% medium-chain triglycerides (70:30 MCT:PFO). An endotoxin challenge was administered to half of the animals in each group at the completion of dietary treatment. All groups demonstrated normal growth throughout the study period. Groups fed MCT and HCO diets demonstrated biochemical essential fatty acid deficiency and decreased IL-6 and TNF-α response to endotoxin challenge. Groups containing PFO had increased inflammatory response to endotoxin challenge, and the addition of MCT to PFO mitigated this inflammatory response. These results suggest that the addition of MCT to PFO formulations may decrease the host response to inflammatory challenge, which may pose potential for optimized PN formulations. Inclusion of MCT in lipid emulsions given with PN formulations may be of use in therapeutic interventions for disease states resulting from chronic inflammation. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Continuous Quadrupole Magnetic Separation of Islets during Digestion Improves Purified Porcine Islet Viability.

    Science.gov (United States)

    Weegman, Bradley P; Kumar Sajja, Venkata Sunil; Suszynski, Thomas M; Rizzari, Michael D; Scott Iii, William E; Kitzmann, Jennifer P; Mueller, Kate R; Hanley, Thomas R; Kennedy, David J; Todd, Paul W; Balamurugan, Appakalai N; Hering, Bernhard J; Papas, Klearchos K

    2016-01-01

    Islet transplantation (ITx) is an emerging and promising therapy for patients with uncontrolled type 1 diabetes. The islet isolation and purification processes require exposure to extended cold ischemia, warm-enzymatic digestion, mechanical agitation, and use of damaging chemicals for density gradient separation (DG), all of which reduce viable islet yield. In this paper, we describe initial proof-of-concept studies exploring quadrupole magnetic separation (QMS) of islets as an alternative to DG to reduce exposure to these harsh conditions. Three porcine pancreata were split into two parts, the splenic lobe (SPL) and the combined connecting/duodenal lobes (CDL), for paired digestions and purifications. Islets in the SPL were preferentially labeled using magnetic microparticles (MMPs) that lodge within the islet microvasculature when infused into the pancreas and were continuously separated from the exocrine tissue by QMS during the collection phase of the digestion process. Unlabeled islets from the CDL were purified by conventional DG. Islets purified by QMS exhibited significantly improved viability (measured by oxygen consumption rate per DNA, p < 0.03) and better morphology relative to control islets. Islet purification by QMS can reduce the detrimental effects of prolonged exposure to toxic enzymes and density gradient solutions and substantially improve islet viability after isolation.

  7. Continuous Quadrupole Magnetic Separation of Islets during Digestion Improves Purified Porcine Islet Viability

    Directory of Open Access Journals (Sweden)

    Bradley P. Weegman

    2016-01-01

    Full Text Available Islet transplantation (ITx is an emerging and promising therapy for patients with uncontrolled type 1 diabetes. The islet isolation and purification processes require exposure to extended cold ischemia, warm-enzymatic digestion, mechanical agitation, and use of damaging chemicals for density gradient separation (DG, all of which reduce viable islet yield. In this paper, we describe initial proof-of-concept studies exploring quadrupole magnetic separation (QMS of islets as an alternative to DG to reduce exposure to these harsh conditions. Three porcine pancreata were split into two parts, the splenic lobe (SPL and the combined connecting/duodenal lobes (CDL, for paired digestions and purifications. Islets in the SPL were preferentially labeled using magnetic microparticles (MMPs that lodge within the islet microvasculature when infused into the pancreas and were continuously separated from the exocrine tissue by QMS during the collection phase of the digestion process. Unlabeled islets from the CDL were purified by conventional DG. Islets purified by QMS exhibited significantly improved viability (measured by oxygen consumption rate per DNA, p<0.03 and better morphology relative to control islets. Islet purification by QMS can reduce the detrimental effects of prolonged exposure to toxic enzymes and density gradient solutions and substantially improve islet viability after isolation.

  8. Optimum design of water supply purifying station in living section of a uranium mine

    International Nuclear Information System (INIS)

    Li Jianjun; Zhang Yu

    2012-01-01

    A design of water supply purifying station in living section of a uranium mine was optimized, and appropriate technique and equipment were chosen based on the raw water quality characteristic, water consumption and change, landform of construction field, etc. After the engineering finished, the circulation is steady-going, the quality of treated water fulfills water standards for drinking water quality. (authors)

  9. Excretory/secretory-products of Echinococcus multilocularis larvae induce apoptosis and tolerogenic properties in dendritic cells in vitro.

    Directory of Open Access Journals (Sweden)

    Justin Komguep Nono

    Full Text Available BACKGROUND: Alveolar echinococcosis, caused by Echinococcus multilocularis larvae, is a chronic disease associated with considerable modulation of the host immune response. Dendritic cells (DC are key effectors in shaping the immune response and among the first cells encountered by the parasite during an infection. Although it is assumed that E.multilocularis, by excretory/secretory (E/S-products, specifically affects DC to deviate immune responses, little information is available on the molecular nature of respective E/S-products and their mode of action. METHODOLOGY/PRINCIPAL FINDINGS: We established cultivation systems for exposing DC to live material from early (oncosphere, chronic (metacestode and late (protoscolex infectious stages. When co-incubated with Echinococcus primary cells, representing the invading oncosphere, or metacestode vesicles, a significant proportion of DC underwent apoptosis and the surviving DC failed to mature. In contrast, DC exposed to protoscoleces upregulated maturation markers and did not undergo apoptosis. After pre-incubation with primary cells and metacestode vesicles, DC showed a strongly impaired ability to be activated by the TLR ligand LPS, which was not observed in DC pre-treated with protoscolex E/S-products. While none of the larvae induced the secretion of pro-inflammatory IL-12p70, the production of immunosuppressive IL-10 was elevated in response to primary cell E/S-products. Finally, upon incubation with DC and naïve T-cells, E/S-products from metacestode vesicles led to a significant expansion of Foxp3+ T cells in vitro. CONCLUSIONS: This is the first report on the induction of apoptosis in DC by cestode E/S-products. Our data indicate that the early infective stage of E. multilocularis is a strong inducer of tolerance in DC, which is most probably important for generating an immunosuppressive environment at an infection phase in which the parasite is highly vulnerable to host attacks. The

  10. Effect of preparation conditions on the characteristics and photocatalytic activity of TiO2/purified diatomite composite photocatalysts

    International Nuclear Information System (INIS)

    Sun, Zhiming; Hu, Zhibo; Yan, Yang; Zheng, Shuilin

    2014-01-01

    Highlights: • TiO 2 /purified diatomite composites were synthesized under different conditions. • The optimum preparation conditions of composites were obtained. • The obtained photocatalyst showed good photocatalytic activity. • The dispersity and grain size of loaded TiO 2 NPs are the critical factors. - Abstract: TiO 2 /purified diatomite composite materials were prepared through a modified hydrolysis-deposition method under low temperature using titanium tetrachloride as precursor combined with a calcination crystallization process. The microstructure and crystalline phases of the obtained composites prepared under different preparation conditions were characterized by high resolution scanning electron microscope (SEM) and X-ray diffraction (XRD), respectively. The photocatalytic performance of TiO 2 /purified diatomite composites was evaluated by Rhodamine B as the target pollutant under UV irradiation, and the optimum preparation conditions of composites were obtained. The TiO 2 crystal form in composites prepared under optimum conditions was anatase, the grain size of which was 34.12 nm. The relationships between structure and property of composite materials were analyzed and discussed. It is indicated that the TiO 2 nanoparticles uniformly dispersed on the surface of diatoms, and the photocatalytic performance of the composite materials was mainly determined by the dispersity and grain size of loaded TiO 2 nanoparticles

  11. Use of ion exchange during preparation of raw materials for production of molybdenum and tungsten of high purify

    International Nuclear Information System (INIS)

    Blokhin, A.A.; Rumyantsev, V.K.; Taushkanov, V.P.; Maksimkov, S.M.; Majorov, D.Yu.; Pak, V.I.

    1988-01-01

    The data on the application of selective ionites for the steep purification of ammonium molybdate and tungstate solutions, are given. It is shown that to purify molybdenum and ammonium tungstate solutions from the impuerities of alkali earth and two- and threevalent transition metals, iminodiacetate ampholites of the ANKB-35 type are the most effective sorbents. To purify from phosphorus, silicon and arsenic impurities composition ionites on the base of hydrated oxides of multivalent metals introduced in the granules of porous cationites should be used. To extract phosphorus, silicon, arsenic impurities from ammonium molybdate and tungstate solutions and tungsten from ammonium molybdate solutions the method of their coprecipitation with iron (3) hydroxide can also be used. The best results on the purification of ammonium molybdate from tungstate provides for the application of structural organomineral ionites as well as weak-basicity anionites of the AN-31 type and its different modifications containing hydroxyl groups along with aminogroups. To purity ammonium tungstate solutions from molybdate a method is developed which transforms molybdenum in the form of thiocomplexes and the following selective sorption by strong-basicity anionites. The data on the quality of molybdenum monocrystals which are taken from the raw material purified using ionites, are given

  12. Soybean plant growth study conducted using purified protein hydrolysate-based fertilizer made from chrome-tanned leather waste.

    Science.gov (United States)

    Pati, Anupama; Chaudhary, Rubina

    2015-12-01

    Leather processing discharges enormous amount of chrome containing leather solid waste which creates a major disposal problem. Chrome-tanned leather solid waste is a complex of collagen and chromium. The presence of chromium limits protein application in fertilizer industry. The purified protein hydrolysate with zero chromium could be used as a nitrogen source for fertilizer formulation. In this study, an attempt has been made to employ purified protein hydrolysate derived from chrome-tanned leather shavings (CTLS) in formulation of fertilizer. The formulated fertilizer (1–3 t ha(-1)) is employed as nitrogen source in production of soybean. Plant growth study demonstrates that formulated fertilizer dosage 3 t ha(-1) produced similar effects of commercial fertilizer-treated plants. Application of formulated fertilizer yielded higher seed in plant than commercial fertilizer.

  13. In vitro and in vivo antioxidant activities of polysaccharide purified from aloe vera (Aloe barbadensis) gel.

    Science.gov (United States)

    Kang, Min-Cheol; Kim, Seo Young; Kim, Yoon Taek; Kim, Eun-A; Lee, Seung-Hong; Ko, Seok-Chun; Wijesinghe, W A J P; Samarakoon, Kalpa W; Kim, Young-Sun; Cho, Jin Hun; Jang, Hyeang-Su; Jeon, You-Jin

    2014-01-01

    The in vitro and in vivo antioxidant potentials of a polysaccharide isolated from aloe vera gel were investigated. Enzymatic extracts were prepared from aloe vera gel by using ten digestive enzymes including five carbohydrases and five proteases. Among them, the highest yield was obtained with the Viscozyme extract and the same extract showed the best radical scavenging activity. An active polysaccharide was purified from the Viscozyme extract using ethanol-added separation and anion exchange chromatography. Purified aloe vera polysaccharide (APS) strongly scavenged radicals including DPPH, hydroxyl and alkyl radicals. In addition, APS showed a protective effect against AAPH-induced oxidative stress and cell death in Vero cells as well as in the in vivo zebrafish model. In this study, it is proved that both the in vitro and in vivo antioxidant potentials of APS could be further utilized in relevant industrial applications. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Composition and Potency Characterization of Mycobacterium avium subsp. paratuberculosis Purified Protein Derivatives.

    Directory of Open Access Journals (Sweden)

    Randal T Capsel

    Full Text Available Mycobacterium avium subsp. paratuberculosis (MAP purified protein derivatives (PPDs are immunologic reagents prepared from cultured filtrates of the type strain. Traditional production consists of floating culture incubation at 37°C, organism inactivation by autoclaving, coarse filtration, and protein precipitation. Three traditional production PPDs were used in this study including lot 9801, which served as a reference and has been used in the field for decades. Alternative production PPDs (0902A and 0902B, in which the autoclaving step was removed, were also analyzed in this study. SDS-PAGE analysis revealed protein smearing in traditional PPDs, but distinct bands were observed in the alternative PPD preparations. Antibody bound distinct protein bands in the alternative PPDs by immunoblot analysis, whereas an immunoreactive smear was observed with the traditional PPDs. Mass spectrometry identified 194 proteins among three PPD lots representing the two different production methods, ten of which were present in all PPDs examined. Selected proteins identified by mass spectrometry were recombinantly expressed and purified from E. coli and evaluated by the guinea pig potency test. Seven recombinant proteins showed greater erythema as compared to the reference PPD lot 9801 in paired guinea pigs and were able to stimulate interferon-gamma production in blood from Johne's positive animals. These results suggest that autoclaving culture suspensions is not a necessary step in PPD production and specific proteins could supplant the PPD antigen for intradermal skin testing procedures and for use as in-vitro assay reagents.

  15. Evaluation of the purified fraction of Wilbrandia (c. f. verticillata for antitumour activity

    Directory of Open Access Journals (Sweden)

    V. S. N. Rao

    1991-01-01

    Full Text Available Cucurbatacins are known to produce cytotoxic and anticancer activities. Two novel norcucurbitacin glucosides (Wvl and Wv2 have recently been isolated from a purified fraction obtained from the rhizome of Wilbrandia verticillata. The present study evaluates the cytotoxic and anti-tumour activities of the norcucurbitacins. We have found a regular cytotoxicity in KB cells (Cy50 = 12µg/ml as well as a significant inhibition in the Walker 256 carcinosarcoma growth (approximately 75%.

  16. Biocompatibility evaluation of HDPE-UHMWPE reinforced β-TCP nanocomposites using highly purified human osteoblast cells.

    Science.gov (United States)

    Shokrgozar, M A; Farokhi, M; Rajaei, F; Bagheri, M H A; Azari, Sh; Ghasemi, I; Mottaghitalab, F; Azadmanesh, K; Radfar, J

    2010-12-15

    Biocompatibility of β-TCP/HDPE-UHMWPE nanocomposite as a new bone substitute material was evaluated by using highly purified human osteoblast cells. Human osteoblast cells were isolated from bone tissue and characterized by immunofluorescence Staining before and after purification using magnetic bead system. Moreover, proliferation, alkaline phosphatase production, cell attachment, calcium deposition, gene expression, and morphology of osteoblast cells on β-TCP/HDPE-UHMWPE nanocomposites were evaluated. The results have shown that the human osteoblast cells were successfully purified and were suitable for subsequent cell culturing process. The high proliferation rate of osteoblast cells on β-TCP/HDPE-UHMWPE nanocomposite confirmed the great biocompatibility of the scaffold. Expression of bone-specific genes was taken place after the cells were incubated in composite extract solutions. Furthermore, osteoblast cells were able to mineralize the matrix next to composite samples. Scanning electron microscopy demonstrated that cells had normal morphology on the scaffold. Thus, these results indicated that the nanosized β-TCP/HDPE-UHMWPE blend composites could be potential scaffold, which is used in bone tissue engineering. Copyright © 2010 Wiley Periodicals, Inc.

  17. Cultured rat and purified human Pneumocystis carinii stimulate intra- but not extracellular free radical production in human neutrophils

    DEFF Research Database (Denmark)

    Jensen, T; Aliouat, E M; Lundgren, B

    1998-01-01

    The production of free radicals in human neutrophils was studied in both Pneumocystis carinii derived from cultures of L2 rat lung epithelial-like cells and Pneumocystis carinii purified from human lung. Using the cytochrome C technique, which selectively measured extracellular superoxide...... generation, hardly any free radical production was observed after stimulation with cultured rat-derived P. carinii. A chemiluminescence technique, which separately measured intra- and extracellular free radical production, was subsequently employed to differentiate the free radical generation....... It was established that 1) P. carinii stimulated intra- but not extracellular free radical production in human neutrophils, 2) opsonized cultured rat-derived P. carinii stimulated human neutrophils to a strong intracellular response of superoxide production, and 3) opsonized P. carinii, purified from human lung also...

  18. Oxidation of eugenol by purified human term placental peroxidase.

    Science.gov (United States)

    Zhang, R; Kulkarni, K A; Kulkarni, A P

    2000-01-01

    The oxidation of eugenol by purified human term placental peroxidase (HTPP) was examined. Spectral analyses indicated that, similar to horseradish peroxidase, HTPP is capable of catalyzing the oxidation of eugenol. The accumulated stable product in the reaction medium due to eugenol oxidation by HTPP was tentatively identified as quinone methide of eugenol (EQM). The EQM formation exhibited a pH optimum of 8.0 and was dependent on incubation time, amount of HTPP and the concentration of both eugenol and hydrogen peroxide. The specific activity of approx 2.8 micromoles of EQM/min/mg protein was observed with different preparations of HTPP. The EQM formation was significantly suppressed by glutathione and ascorbic acid. The classical peroxidase inhibitors viz. potassium cyanide and sodium azide blocked the reaction in a concentration manner. Collectively, the results suggest that eugenol may undergo peroxidative metabolism in human placenta. Copyright 2000 Harcourt Publishers Ltd.

  19. An evaluation of purified Salmonella Typhi protein antigens for the serological diagnosis of acute typhoid fever

    NARCIS (Netherlands)

    Tran Vu Thieu, Nga; Trinh van, Tan; Tran Tuan, Anh; Klemm, Elizabeth J.; Nguyen Ngoc Minh, Chau; Voong Vinh, Phat; Pham Thanh, Duy; Ho Ngoc Dan, Thanh; Pham Duc, Trung; Langat, Pinky; Martin, Laura B.; Galan, Jorge; Liang, Li; Felgner, Philip L.; Davies, D. Huw; de Jong, Hanna K.; Maude, Rapeephan R.; Fukushima, Masako; Wijedoru, Lalith; Ghose, Aniruddha; Samad, Rasheda; Dondorp, Arjen M.; Faiz, Abul; Darton, Thomas C.; Pollard, Andrew J.; Thwaites, Guy E.; Dougan, Gordon; Parry, Christopher M.; Baker, Stephen

    2017-01-01

    The diagnosis of typhoid fever is a challenge. Aiming to develop a typhoid diagnostic we measured antibody responses against Salmonella Typhi (S. Typhi) protein antigens and the Vi polysaccharide in a cohort of Bangladeshi febrile patients. IgM against 12 purified antigens and the Vi polysaccharide

  20. Phospholipid analysis and fractional reconstitution of the ice nucleation protein activity purified from Escherichia coli overexpressing the inaZ gene of Pseudomonas syringae.

    Science.gov (United States)

    Palaiomylitou, M A; Kalimanis, A; Koukkou, A I; Drainas, C; Anastassopoulos, E; Panopoulos, N J; Ekateriniadou, L V; Kyriakidis, D A

    1998-08-01

    Ice nucleation protein was partially purified from the membrane fraction of E. coli carrying inaZ from Pseudomonas syringae. The ice nucleation protein was totally localized in the bacterial envelope and was extracted by either salt (0.25 M NH4Cl) or the nonionic detergent Tween 20. The extracted protein was partially purified by sequential passage through DEAE-52 cellulose and Sephacryl-S400 columns. The activity of the purified protein was lost after treatment with phospholipase C, and its activity was subsequently restored by addition of the naturally occurring lipid phosphatidylethanolamine. These results suggest that ice nucleation proteins have a requirement for lipids that reconstitute a physiological hydrophobic environment similar to the one existing in vivo, to attain and maintain a structure that enables ice catalysis. Copyright 1998 Academic Press.

  1. The purified ATPase from chromaffin granule membranes is an anion-dependent proton pump.

    Science.gov (United States)

    Moriyama, Y; Nelson, N

    1987-07-05

    The proton-ATPase of chromaffin granules was purified so as to maintain its proton-pumping activity when reconstituted into phospholipid vesicles. The purification procedure involved solubilization with polyoxyethylene 9 lauryl ether, hydroxylapatite column, precipitation by ammonium sulfate, and glycerol gradient centrifugation. The protease inhibitor mixture used in previous studies inhibited the proton-pumping activity of the enzyme; therefore, the protein was stabilized by pepstatin A and leupeptin. The enzyme was purified at least 50-fold with respect to both ATPase and proton-pumping activity. The ATP-dependent proton uptake activity of the reconstituted enzyme was absolutely dependent on the presence of Cl- or Br- outside the vesicles, whereas sulfate, acetate, formate, nitrate, and thiocyanate were inhibitory. Sulfate inhibition seems to be due to competition with Cl- on the anion-binding site outside the vesicles, whereas nitrate and thiocyanate inhibited only from the internal side. As with the inhibition by N-ethylmaleimide, the proton-pumping activity was much more sensitive to nitrate than the ATPase activity. About 20 mM nitrate were sufficient for 90% inhibition of the proton-pumping activity while 100 mM inhibited only 50% of the ATPase activity both in situ and in the reconstituted enzyme. The possible regulatory effect of anions on the ATP-dependent proton uptake in secretory granules is discussed.

  2. Neutralisation of the anti-coagulant effects of heparin by histones in blood plasma and purified systems.

    Science.gov (United States)

    Longstaff, Colin; Hogwood, John; Gray, Elaine; Komorowicz, Erzsebet; Varjú, Imre; Varga, Zoltán; Kolev, Krasimir

    2016-03-01

    Neutrophil extracellular traps (NETs) composed primarily of DNA and histones are a link between infection, inflammation and coagulation. NETs promote coagulation and approaches to destabilise NETs have been explored to reduce thrombosis and treat sepsis. Heparinoids bind histones and we report quantitative studies in plasma and purified systems to better understand physiological consequences. Unfractionated heparin (UFH) was investigated by activated partial thromboplastin time (APTT) and alongside low-molecular-weight heparins (LMWH) in purified systems with thrombin or factor Xa (FXa) and antithrombin (AT) to measure the sensitivity of UFH or LMWH to histones. A method was developed to assess the effectiveness of DNA and non-anticoagulant heparinoids as anti-histones. Histones effectively neutralised UFH, the IC50 value for neutralisation of 0.2 IU/ml UFH was 1.8 µg/ml histones in APTT and 4.6 µg/ml against 0.6 IU/ml UFH in a purified system. Histones also inhibited the activities of LMWHs with thrombin (IC50 6.1 and 11.0 µg/ml histones, for different LMWHs) or FXa (IC50 7.8 and 7.0 µg/ml histones). Direct interactions of UFH and LMWH with DNA and histones were explored by surface plasmon resonance, while rheology studies showed complex effects of histones, UFH and LMWH on clot resilience. A conclusion from these studies is that anticoagulation by UFH and LMWH will be compromised by high affinity binding to circulating histones even in the presence of DNA. A complete understanding of the effects of histones, DNA and heparins on the haemostatic system must include an appreciation of direct effects on fibrin and clot structure.

  3. Photoaffinity labelling of a small protein component of a purified (Na+-K+)ATPase

    International Nuclear Information System (INIS)

    Rogers, T.B.; Lazdunski, M.

    1979-01-01

    Studies have been carried out on the photoaffinity labelling of the (Na + -K + )ATPase from the electric organ of Electrophorus electricus. The aims were to see if different photoaffinity labels of the ouabain binding site, are capable of labelling a small protein component and to know if there is a small protein component, in addition to the major protein chains with molecular weights in the regions of 100 000 and 50 000, which is present in other purified (Na + -K + )ATPase preparations. (Auth.)

  4. Dynamics of spherical metallic particles in cylinder electrostatic separators/purifiers.

    Science.gov (United States)

    Lu, Hong-Zhou; Li, Jia; Guo, Jie; Xu, Zhen-Ming

    2008-08-15

    This paper presents a theoretical analysis of the dynamics of spherical metallic particles in electrostatic separators/purifiers (ESPs). The particle equations of motion are numerically solved in two dimensions using a computational algorithm. The ESPs consist of a pair of conductor cylinder electrodes. The upper cylinder is energized by HVdc, while the lower one is grounded and fixed horizontally on a revolvable axis. Some phenomena and aspects of separation process are explained and depicted including lifting off, impact, "motion collapse" and "sudden bouncing". The results reveal that the several phenomena depend on initial position, radius and density of the particle, curvature of the cylinder electrodes, distance between the electrodes and amplitude of the applied voltage. Optimization of the parameters is presented in order to get better separation/purification processes.

  5. Cysticercosis/taeniasis in Asia and the Pacific.

    Science.gov (United States)

    Ito, Akira; Wandra, Toni; Yamasaki, Hiroshi; Nakao, Minoru; Sako, Yasuhito; Nakaya, Kazuhiro; Margono, Sri S; Suroso, Thomas; Gauci, Charles; Lightowlers, Marshall W

    2004-01-01

    Three taeniid tapeworms infect humans in Asia and the Pacific: Taenia solim, Taenia saginata, and Taenia asiatica. Although there is continuing debate about the definition of a new species, phylogenetic analyses of these parasites have provided multiple lines of evidence that T. asiatica is an independent species and the sister species of T. saginata. Here we review briefly the morphology, pathology, molecular biology, distribution and control options of taeniasis/cysticercosis in Asia and the Pacific and comment on the potential role which dogs may play in the transmission of T. solium. Special attention is focused on Indonesia: taeniasis caused by T. asiatica in North Sumatra, taeniasis/cysticercosis of T. solium and taeniasis of T. saginata in Bali, and taeniasis/cysticercosis of T. solium in Papua (formerly Irian Jaya). Issues relating to the spread of taeniasis/cysticercosis caused by T. solium in Papua New Guinea are highlighted, since serological evidence suggests that cysticercosis occurs among the local residents. The use of modern techniques for detection of taeniasis in humans and cysticercosis in humans, pigs and dogs, with the possible adoption of new control measures will provide a better understanding of the epidemiology of taeniasis/cysticercosis in Asia and the Pacific and lead to improved control of zoonotic and simultaneously meat-borne disease transmission.

  6. Structural characterization of the thermally-tolerant pectin methylesterase purified from Citrus sinensis fruit and its gene sequence

    Science.gov (United States)

    Despite the longstanding importance for the thermally-tolerant pectin methylesterase (TT-PME) activity in citrus juice processing and product quality, unequivocal identification of the protein and its corresponding gene has remained elusive. We purified TT-PME from sweet orange [Citrus sinensis (L.)...

  7. Regulated eukaryotic DNA replication origin firing with purified proteins.

    Science.gov (United States)

    Yeeles, Joseph T P; Deegan, Tom D; Janska, Agnieszka; Early, Anne; Diffley, John F X

    2015-03-26

    Eukaryotic cells initiate DNA replication from multiple origins, which must be tightly regulated to promote precise genome duplication in every cell cycle. To accomplish this, initiation is partitioned into two temporally discrete steps: a double hexameric minichromosome maintenance (MCM) complex is first loaded at replication origins during G1 phase, and then converted to the active CMG (Cdc45-MCM-GINS) helicase during S phase. Here we describe the reconstitution of budding yeast DNA replication initiation with 16 purified replication factors, made from 42 polypeptides. Origin-dependent initiation recapitulates regulation seen in vivo. Cyclin-dependent kinase (CDK) inhibits MCM loading by phosphorylating the origin recognition complex (ORC) and promotes CMG formation by phosphorylating Sld2 and Sld3. Dbf4-dependent kinase (DDK) promotes replication by phosphorylating MCM, and can act either before or after CDK. These experiments define the minimum complement of proteins, protein kinase substrates and co-factors required for regulated eukaryotic DNA replication.

  8. Neuropeptide Y binding sites in rat brain identified with purified neuropeptide Y-I125

    International Nuclear Information System (INIS)

    Walker, M.W.; Miller, R.J.

    1986-01-01

    Neuropeptide Y (NPY) is a widely distributed neuronally localized peptide with 36 amino acids, 5 of which are tyrosines. The authors wished to investigate the properties of specific receptors for NPY. They therefore labeled the tyrosines with I125 using chloramine T and then purified the peptide using HPLC. A single mono-iodinated species of NPY which yielded > 85% specific binding in rat forebrain synaptosomes was selected as the ligand for all subsequent experiments. A time course of binding showed that equilibrium conditions were reached in 60 minutes at 21 0 C. Scatchard plots revealed a single class of binding sites with a Kd and a Bmax of 3 x 10-10 M and 28 pmol/mg, respectively. Competition binding with unlabeled NPY showed 50% displacement of bound ligand at 1 x 10-10 M NPY. Competition binding with rat pancreatic polypeptide (RPP), a homologous peptide possessing little NPY-like activity, showed 50% displacement of bound ligand at 2 x 10 -7 M RPP. No binding was observed on F-11 or PC12 neuronal cell lines, or on HSWP fibroblast cells. They conclude that NPY-I125 purified to homogeneity with HPLC is a highly selective ligand for NPY receptor sites. They are currently investigating such sites in brain, gut, and other tissues

  9. Role of Purified Anthocyanins in Improving Cardiometabolic Risk Factors in Chinese Men and Women with Prediabetes or Early Untreated Diabetes-A Randomized Controlled Trial.

    Science.gov (United States)

    Yang, Liping; Ling, Wenhua; Yang, Yan; Chen, Yuming; Tian, Zezhong; Du, Zhicheng; Chen, Jianying; Xie, Yuanling; Liu, Zhaomin; Yang, Lili

    2017-10-10

    Objective: In vitro and animal studies suggest that purified anthocyanins have favorable effects on metabolic profiles, but clinical trials have reported inconsistent findings. Furthermore, no study has been specifically conducted among individuals with prediabetes. The aim of this study was to investigate whether purified anthocyanins could improve cardiometabolic risk factors in Chinese adults with early untreated hyperglycemia. Research Design and Methods: This was a 12-week randomized, double-blind, placebo-controlled trial. A total of 160 participants aged 40-75 years with prediabetes or early untreated diabetes were randomly allocated to receive either purified anthocyanins (320 mg/day, n = 80) or placebo ( n = 80) of identical appearance. A three-hour oral glucose tolerance test (OGTT) was performed, and cardiometabolic biomarkers (glycated hemoglobin A1c (HbA1c), fasting and postprandial glucose, insulin, C-peptide, and lipids) were measured at baseline and at the end of the trial. Results: A total of 138 subjects completed the protocol. Compared with placebo, purified anthocyanins moderately reduced HbA1c (-0.14%, 95% CI: -0.23~-0.04%; p = 0.005), low-density lipoprotein-c (LDL-c) (-0.2 mmol/L, 95% CI: -0.38~-0.01, p = 0.04), apolipoprotein A-1 (apo A1) (0.09 g/L, 95% CI: 0.02~0.17; p = 0.02), and apolipoprotein B (apo B) (-0.07 g/L, 95% CI: -0.13~-0.01; p = 0.01) according to intention-to-treat analysis. Subgroup analyses suggested that purified anthocyanins were more effective at improving glycemic control, insulin sensitivity, and lipids among patients with elevated metabolic markers. Conclusions: The 12-week randomized controlled trials (RCT) in Chinese adults with prediabetes or early untreated diabetes indicated that purified anthocyanins favorably affected glycemic control and lipid profile. Future studies of a longer duration that explore the dose-response relationship among patients with cardiometabolic disorders are needed to confirm our findings.

  10. Removal of Bound Triton X-100 from Purified Bovine Heart Cytochrome bc1

    OpenAIRE

    Varhač, Rastislav; Robinson, Neal C.; Musatov, Andrej

    2009-01-01

    Cytochrome bc1 isolated from Triton X-100 solubilized mitochondrial membranes contains up to 120 nmol of Triton X-100 bound per nmol of the enzyme. Purified cytochrome bc1 is fully active; however, protein bound Triton X-100 significantly interferes with structural studies of the enzyme. Removal of Triton X-100 bound to bovine cytochrome bc1 was accomplished by incubation with Bio-Beads SM-2 in presence of sodium cholate. Sodium cholate is critical since it does not interfere with the adsorpt...

  11. Studying placental transfer of highly purified non-dioxin-like PCBs in two models of the placental barrier

    DEFF Research Database (Denmark)

    Correia Carreira, S; Cartwright, L; Mathiesen, L

    2011-01-01

    Currently, toxicology and toxicokinetics of purified non-dioxin-like polychlorinated biphenyls (NDL-PCBs) are poorly characterised. Transplacental kinetics of NDL-PCBs can be studied in a variety of models, but careful validation of each model is crucial. We aimed to develop a standard operating...

  12. Production of rabbit antibodies against purified Glucose oxidase

    Directory of Open Access Journals (Sweden)

    Muhammad Anjum Zia

    2012-02-01

    Full Text Available Glucose oxidase is an active oxygen species generating enzyme produced from Aspergillus niger grown in submerged fermentation. Disintegration of the mycelium resulted in high glucose oxidase activity that was subjected to ammonium sulfate precipitation at 60-85% saturation rates that resulted to 6.14 U mg -1 specific activity. Purification of enzyme by anion exchange column (DEAE-Cellulose resulted into 22.53 U mg-1 specific activity and 10.27 fold purification. This was applied to sephadex G-200 column for gel filtration chromatography. It was observed that enzyme achieved 59.37 U mg-1of specific activity with 27.08 fold purity and 64.36% recovery. Purified glucose oxidase was injected into rabbits through intravenous route, to raise the glucose oxidase antibodies. After 30 days incubation period, the rabbits were slaughtered and serum was separated from blood. The antibodies were isolated by ammonium sulfate precipitation and confirmed by agar gel precipitation test. This could be a convenient and low cost alternate assay for the estimation of glucose oxidase in biological fluids. Moreover, such antibodies against the said enzyme could be used in various therapeutic and diagnostic applications.

  13. Can Phlorotannins Purified Extracts Constitute a Novel Pharmacological Alternative for Microbial Infections with Associated Inflammatory Conditions?

    Science.gov (United States)

    Lopes, Graciliana; Sousa, Carla; Silva, Luís R.; Pinto, Eugénia; Andrade, Paula B.; Bernardo, João; Mouga, Teresa; Valentão, Patrícia

    2012-01-01

    Bacterial and fungal infections and the emerging multidrug resistance are driving interest in fighting these microorganisms with natural products, which have generally been considered complementary to pharmacological therapies. Phlorotannins are polyphenols restricted to brown seaweeds, recognized for their biological capacity. This study represents the first research on the antibacterial, antifungal, anti-inflammatory and antioxidant activity of phlorotannins purified extracts, which were obtained from ten dominant brown seaweeds of the occidental Portuguese coast. Phlorotannins content was determined by the specific dimethoxybenzaldehyde (DMBA) method and a yield between 75 and 969 mg/Kg phloroglucinol units (dry matter) was obtained. Fucus spiralis ranked first, followed by three Cystoseira species. The anti-inflammatory potential of the purified extracts was assessed via inhibitory effect on nitric oxide (NO) production by lipopolysaccharide-stimulated RAW 264.7 macrophage cells, Cystoseira tamariscifolia being the one showing promising activity for the treatment of inflammation. NO scavenging ability was also addressed in cell free systems, F. spiralis being the species with highest capacity. The antimicrobial potential of the extracts was checked against five Gram-positive and four Gram-negative bacteria and three fungi strains, that commonly colonize skin and mucosa and are responsible for food contamination. The different extracts were more effective against Gram-positive bacteria, Staphylococcus epidermidis being the most susceptible species. Concerning antifungal activity, Trichophyton rubrum was the most sensitive species. Although the molecular mechanisms underlying these properties remain poorly understood, the results obtained turn phlorotannins purified extracts a novel and potent pharmacological alternative for the treatment of a wide range of microbial infections, which usually also present an inflammatory component. In addition to the biological

  14. Bovine pancreatic trypsin inhibitor immobilized onto sepharose as a new strategy to purify a thermostable alkaline peptidase from cobia (Rachycentron canadum) processing waste.

    Science.gov (United States)

    França, Renata Cristina da Penha; Assis, Caio Rodrigo Dias; Santos, Juliana Ferreira; Torquato, Ricardo José Soares; Tanaka, Aparecida Sadae; Hirata, Izaura Yoshico; Assis, Diego Magno; Juliano, Maria Aparecida; Cavalli, Ronaldo Olivera; Carvalho, Luiz Bezerra de; Bezerra, Ranilson Souza

    2016-10-15

    A thermostable alkaline peptidase was purified from the processing waste of cobia (Rachycentron canadum) using bovine pancreatic trypsin inhibitor (BPTI) immobilized onto Sepharose. The purified enzyme had an apparent molecular mass of 24kDa by both sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry. Its optimal temperature and pH were 50°C and 8.5, respectively. The enzyme was thermostable until 55°C and its activity was strongly inhibited by the classic trypsin inhibitors N-ρ-tosyl-l-lysine chloromethyl ketone (TLCK) and benzamidine. BPTI column allowed at least 15 assays without loss of efficacy. The purified enzyme was identified as a trypsin and the N-terminal amino acid sequence of this trypsin was IVGGYECTPHSQAHQVSLNSGYHFC, which was highly homologous to trypsin from cold water fish species. Using Nα-benzoyl-dl-arginine ρ-nitroanilide hydrochloride (BApNA) as substrate, the apparent km value of the purified trypsin was 0.38mM, kcat value was 3.14s(-1), and kcat/km was 8.26s(-1)mM(-1). The catalytic proficiency of the purified enzyme was 2.75×10(12)M(-1) showing higher affinity for the substrate at the transition state than other fish trypsin. The activation energy (AE) of the BApNA hydrolysis catalyzed by this enzyme was estimated to be 11.93kcalmol(-1) while the resulting rate enhancement of this reaction was found to be approximately in a range from 10(9) to 10(10)-fold evidencing its efficiency in comparison to other trypsin. This new purification strategy showed to be appropriate to obtain an alkaline peptidase from cobia processing waste with high purification degree. According with N-terminal homology and kinetic parameters, R. canadum trypsin may gathers desirable properties of psychrophilic and thermostable enzymes. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Practical screening of purified cellobiohydrolases and endoglucanases with α-cellulose and specification of hydrodynamics

    Directory of Open Access Journals (Sweden)

    Jäger Gernot

    2010-08-01

    Full Text Available Abstract Background It is important to generate biofuels and society must be weaned from its dependency on fossil fuels. In order to produce biofuels, lignocellulose is pretreated and the resulting cellulose is hydrolyzed by cellulases such as cellobiohydrolases (CBH and endoglucanases (EG. Until now, the biofuel industry has usually applied impractical celluloses to screen for cellulases capable of degrading naturally occurring, insoluble cellulose. This study investigates how these cellulases adsorb and hydrolyze insoluble α-cellulose − considered to be a more practical substrate which mimics the alkaline-pretreated biomass used in biorefineries. Moreover, this study investigates how hydrodynamics affects cellulase adsorption and activity onto α-cellulose. Results First, the cellulases CBH I, CBH II, EG I and EG II were purified from Trichoderma reesei and CBH I and EG I were utilized in order to study and model the adsorption isotherms (Langmuir and kinetics (pseudo-first-order. Second, the adsorption kinetics and cellulase activities were studied under different hydrodynamic conditions, including liquid mixing and particle suspension. Third, in order to compare α-cellulose with three typically used celluloses, the exact cellulase activities towards all four substrates were measured. It was found that, using α-cellulose, the adsorption models fitted to the experimental data and yielded parameters comparable to those for filter paper. Moreover, it was determined that higher shaking frequencies clearly improved the adsorption of cellulases onto α-cellulose and thus bolstered their activity. Complete suspension of α-cellulose particles was the optimal operating condition in order to ensure efficient cellulase adsorption and activity. Finally, all four purified cellulases displayed comparable activities only on insoluble α-cellulose. Conclusions α-Cellulose is an excellent substrate to screen for CBHs and EGs. This current investigation

  16. Characterization of a protein kinase activity associated with purified capsids of the granulosis virus infecting Plodia interpunctella.

    Science.gov (United States)

    Wilson, M E; Consigli, R A

    1985-06-01

    A cyclic-nucleotide independent protein kinase activity has been demonstrated in highly purified preparations of the granulosis virus infecting the Indian meal moth, Plodia interpunctella. A divalent cation was required for activity. Manganese was the preferred cation and a pH of 8.0 resulted in optimal incorporation of 32P radiolabel into acid-precipitable protein. Although both ATP and GTP could serve as phosphate donors, ATP was utilized more efficiently by the enzyme. The kinase activity was localized to purified capsids; and the basic, internal core protein, VP12, was found to be the predominant viral acceptor. Histones and protamine sulfate could also serve as acceptors for the capsid-associated kinase activity. Using acid hydrolysis and phosphoamino acid analysis of phosphorylated nucleocapsid protein and nuclear magnetic resonance of phosphorylated VP12, it was determined that the enzyme catalyzes the transfer of phosphate to both serine and arginine residues of acceptor proteins. We believe this kinase activity may play a significant role in the viral replication cycle.

  17. [Digestive utilization of purified cellulose in the rainbow trout (Salmo gairdneri) and the common carp (Cyprinus carpio)].

    Science.gov (United States)

    Bergot, F

    1981-01-01

    A semi-purified diet containing 22 p. 100 of a wood cellulose extract without lignin but still containing 22 p. 100 of hemicelluloses was distributed for one month to rainbow trout and common carp reared at 17 and 20 degrees C, respectively. The digestibility of the main dietary constituents was determined by an indirect method using chrome oxide as an inert tracer. The feces were recovered by a continuous automatic collector which rapidly removed them from the water, minimizing alteration by leaching. The cellulose content was estimated by the Weende (crude fiber) and the Van Soest (neutral detergent fiber and acid detergent fiber) methods. The digestibility coefficients obtained for trout as well as for carp indicate that cellulose and hemicelluloses were not digested. In both species, volatile fatty acid concentration in the different segments of the digestive tract was low (less than 10 mM/l). These results lead us to suggest that trout and carp cannot degrade purified cellulose.

  18. Effectiveness of air purifier on health outcomes and indoor particles in homes of children with allergic diseases in Fresno, California: A pilot study.

    Science.gov (United States)

    Park, Hye-Kyung; Cheng, Kai-Chung; Tetteh, Afua O; Hildemann, Lynn M; Nadeau, Kari C

    2017-05-01

    Epidemiologic studies indicate that indoor air pollution is correlated with morbidity caused by allergic diseases. We evaluated the effectiveness of reducing the levels of indoor fine particulate matter <2.5 micrometer diameter (PM 2.5 ) in Fresno, California using air purifiers on health outcomes in children with asthma and/or allergic rhinitis. The active group (with air purifiers) and the control group consisted of eight houses each. Air purifiers were installed in the living rooms and bedrooms of the subjects in the active group during the entire 12-week study duration. Childhood asthma control test, peak flow rate monitoring, and nasal symptom scores were evaluated at weeks 0, 6, and 12. At 12 weeks, the active group showed a trend toward an improvement of childhood asthma control test scores and mean evening peak flow rates, whereas the control group showed deterioration in the same measures. Total and daytime nasal symptoms scores significantly reduced in the active group (p = 0.001 and p = 0.011, respectively). The average indoor PM 2.5 concentrations reduced by 43% (7.42 to 4.28 μg/m 3 ) in the active group (p = 0.001). Intervention with air purifiers reduces indoor PM 2.5 levels with significant improvements in nasal symptoms in children with allergic rhinitis in Fresno.

  19. Recombinant Passenger Proteins Can Be Conveniently Purified by One-Step Affinity Chromatography.

    Science.gov (United States)

    Wang, Hua-zhen; Chu, Zhi-zhan; Chen, Chang-chao; Cao, Ao-cheng; Tong, Xin; Ouyang, Can-bin; Yuan, Qi-hang; Wang, Mi-nan; Wu, Zhong-kun; Wang, Hai-hong; Wang, Sheng-bin

    2015-01-01

    Fusion tag is one of the best available tools to date for enhancement of the solubility or improvement of the expression level of recombinant proteins in Escherichia coli. Typically, two consecutive affinity purification steps are often necessitated for the purification of passenger proteins. As a fusion tag, acyl carrier protein (ACP) could greatly increase the soluble expression level of Glucokinase (GlcK), α-Amylase (Amy) and GFP. When fusion protein ACP-G2-GlcK-Histag and ACP-G2-Amy-Histag, in which a protease TEV recognition site was inserted between the fusion tag and passenger protein, were coexpressed with protease TEV respectively in E. coli, the efficient intracellular processing of fusion proteins was achieved. The resulting passenger protein GlcK-Histag and Amy-Histag accumulated predominantly in a soluble form, and could be conveniently purified by one-step Ni-chelating chromatography. However, the fusion protein ACP-GFP-Histag was processed incompletely by the protease TEV coexpressed in vivo, and a large portion of the resulting target protein GFP-Histag aggregated in insoluble form, indicating that the intracellular processing may affect the solubility of cleaved passenger protein. In this context, the soluble fusion protein ACP-GFP-Histag, contained in the supernatant of E. coli cell lysate, was directly subjected to cleavage in vitro by mixing it with the clarified cell lysate of E. coli overexpressing protease TEV. Consequently, the resulting target protein GFP-Histag could accumulate predominantly in a soluble form, and be purified conveniently by one-step Ni-chelating chromatography. The approaches presented here greatly simplify the purification process of passenger proteins, and eliminate the use of large amounts of pure site-specific proteases.

  20. Effect of preparation conditions on the characteristics and photocatalytic activity of TiO{sub 2}/purified diatomite composite photocatalysts

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Zhiming, E-mail: szmcumtb@hotmail.com; Hu, Zhibo; Yan, Yang; Zheng, Shuilin, E-mail: shuilinzh@sina.com

    2014-09-30

    Highlights: • TiO{sub 2}/purified diatomite composites were synthesized under different conditions. • The optimum preparation conditions of composites were obtained. • The obtained photocatalyst showed good photocatalytic activity. • The dispersity and grain size of loaded TiO{sub 2} NPs are the critical factors. - Abstract: TiO{sub 2}/purified diatomite composite materials were prepared through a modified hydrolysis-deposition method under low temperature using titanium tetrachloride as precursor combined with a calcination crystallization process. The microstructure and crystalline phases of the obtained composites prepared under different preparation conditions were characterized by high resolution scanning electron microscope (SEM) and X-ray diffraction (XRD), respectively. The photocatalytic performance of TiO{sub 2}/purified diatomite composites was evaluated by Rhodamine B as the target pollutant under UV irradiation, and the optimum preparation conditions of composites were obtained. The TiO{sub 2} crystal form in composites prepared under optimum conditions was anatase, the grain size of which was 34.12 nm. The relationships between structure and property of composite materials were analyzed and discussed. It is indicated that the TiO{sub 2} nanoparticles uniformly dispersed on the surface of diatoms, and the photocatalytic performance of the composite materials was mainly determined by the dispersity and grain size of loaded TiO{sub 2} nanoparticles.

  1. Efficacy of Albendazole-Chitosan Microsphere-based Treatment for Alveolar Echinococcosis in Mice.

    Directory of Open Access Journals (Sweden)

    Maitiseyiti Abulaihaiti

    Full Text Available This study aimed to investigate the pharmacology and anti-parasitic efficacy of albendazole-chitosan microspheres (ABZ-CS-MPs for established intraperitoneal infections of Echinococcus multilocularis metacestodes in an experimental murine model. Male outbred Kunming mice infected with E. multilocularis Metacestodes were administered with three ABZ formulations, namely, ABZ-CS-MPs, Liposome-Albendazole (L-ABZ, and albendazole tablet (ABZ-T. Each of the ABZ formulations was given orally at three different doses of 37.5, 75, and 150 mg/kg, three times a week for 12 weeks postinfection. After administering the drugs, we monitored the pharmacological performance and anti-parasitic efficacy of ABZ-CS-MPs compared with L-ABZ, and ABZ-T treated mice. ABZ-CS-MPs reduced the weight of tissues containing E. multilocularis metacestodes most effectively compared with the ABZ-T group and untreated controls. Metacestode grown was Highly suppressed during treatment with ABZ-CS-MPs. Significantly higher plasma levels of ABZ metabolites were measured in mice treated with ABZ-CS-MPs or L-ABZ compared with ABZ-T. In particular, enhanced ABZ-sulfoxide concentration profiles were observed in the mice given 150 mg/kg of ABZ-CS-MPs, but not in the mice treated with L-ABZ. Histological examination showed that damages caused disorganization of both the germinal and laminated layers of liver hyatid cysts, demolishing their characteristic structures after treatment with ABZ-CS-MPs or L-ABZ. Over time, ABZ-CS-MPs treatment induced a shift from Th2-dominant to Th1-dominant immune response. CS-MPs As a new carrier exhibited improved absorption and increased bioavailability of ABZ in the treatment of E. multilocularis infections in mice.

  2. Efficacy of Albendazole-Chitosan Microsphere-based Treatment for Alveolar Echinococcosis in Mice.

    Science.gov (United States)

    Abulaihaiti, Maitiseyiti; Wu, Xiang-Wei; Qiao, Lei; Lv, Hai-Long; Zhang, Hong-Wei; Aduwayi, Nasrul; Wang, Yan-Jie; Wang, Xin-Chun; Peng, Xin-Yu

    2015-01-01

    This study aimed to investigate the pharmacology and anti-parasitic efficacy of albendazole-chitosan microspheres (ABZ-CS-MPs) for established intraperitoneal infections of Echinococcus multilocularis metacestodes in an experimental murine model. Male outbred Kunming mice infected with E. multilocularis Metacestodes were administered with three ABZ formulations, namely, ABZ-CS-MPs, Liposome-Albendazole (L-ABZ), and albendazole tablet (ABZ-T). Each of the ABZ formulations was given orally at three different doses of 37.5, 75, and 150 mg/kg, three times a week for 12 weeks postinfection. After administering the drugs, we monitored the pharmacological performance and anti-parasitic efficacy of ABZ-CS-MPs compared with L-ABZ, and ABZ-T treated mice. ABZ-CS-MPs reduced the weight of tissues containing E. multilocularis metacestodes most effectively compared with the ABZ-T group and untreated controls. Metacestode grown was Highly suppressed during treatment with ABZ-CS-MPs. Significantly higher plasma levels of ABZ metabolites were measured in mice treated with ABZ-CS-MPs or L-ABZ compared with ABZ-T. In particular, enhanced ABZ-sulfoxide concentration profiles were observed in the mice given 150 mg/kg of ABZ-CS-MPs, but not in the mice treated with L-ABZ. Histological examination showed that damages caused disorganization of both the germinal and laminated layers of liver hyatid cysts, demolishing their characteristic structures after treatment with ABZ-CS-MPs or L-ABZ. Over time, ABZ-CS-MPs treatment induced a shift from Th2-dominant to Th1-dominant immune response. CS-MPs As a new carrier exhibited improved absorption and increased bioavailability of ABZ in the treatment of E. multilocularis infections in mice.

  3. PRESERVATIVE POTENTIAL OF PURIFIED BACTERIOCIN PRODUCED FROM BREVIBACILLUS BORSTELENSIS AG1 ISOLATED FROM MARCHA – A TRADITIONAL WINE STARTER CULTURE CAKE IN TOMATO PASTE

    OpenAIRE

    Anupama Gupta; Nivedita Sharma; Neha Gautam

    2015-01-01

    Purified bacteriocin produced from Brevibacillus borstelensis AG1 isolated from Marcha a local wine starter herbal cake, was used to enhance the shelf life of tomato paste. Preservative effect of purified bacteriocin was studied for nine days in tomato paste inoculated with food borne pathogens and was compared to commercial biopreservative – nisin and chemical preservative – sodium benzoate. The indicator strains i.e. Listeria monocytogenes MTCC839, Bacillus subtilis CRI and Clostridium perf...

  4. Requirements for growth and IL-10 expression of highly purified human T regulatory cells

    OpenAIRE

    Bonacci, Benedetta; Edwards, Brandon; Jia, Shuang; Williams, Calvin; Hessner, Martin J.; Gauld, Stephen; Verbsky, James

    2012-01-01

    Human regulatory T cells (TR) cells have potential for the treatment of a variety of immune mediated diseases but the anergic phenotype of these cells makes them difficult to expand in vitro. We have examined the requirements for growth and cytokine expression from highly purified human TR cells, and correlated these findings with the signal transduction events of these cells. We demonstrate that these cells do not proliferate or secrete IL-10 even in the presence of high doses of IL-2. Stimu...

  5. Specific proliferative response of human lymphocytes to purified soluble antigens from Plasmodium falciparum in vitro cultures and to antigens from malaria patients' sera

    DEFF Research Database (Denmark)

    Bygbjerg, I C; Jepsen, S; Theander, T G

    1985-01-01

    Antigens of Plasmodium falciparum, in supernatants of in vitro cultures of the parasite were affinity purified on columns prepared with the IgG fraction of the serum of an immune individual. The purified antigens induced proliferation of lymphocytes from persons who had recently had malaria....... The responses were strongest with lymphocytes from individuals infected with falciparum and ovale malaria; vivax malaria infections induced a lower level of response and lymphocytes of unsensitized individuals were little affected. Lymphocytes from unsensitized individuals did not respond to the affinity...

  6. Production of Purified CasRNPs for Efficacious Genome Editing.

    Science.gov (United States)

    Lingeman, Emily; Jeans, Chris; Corn, Jacob E

    2017-10-02

    CRISPR-Cas systems have been harnessed as modular genome editing reagents for functional genomics and show promise to cure genetic diseases. Directed by a guide RNA, a Cas effector introduces a double stranded break in DNA and host cell DNA repair leads to the introduction of errors (e.g., to knockout a gene) or a programmed change. Introduction of a Cas effector and guide RNA as a purified Cas ribonucleoprotein complex (CasRNP) has recently emerged as a powerful approach to alter cell types and organisms. Not only does CasRNP editing exhibit increased efficacy and specificity, it avoids optimization and iteration of species-specific factors such as codon usage, promoters, and terminators. CasRNP editing has been rapidly adopted for research use in many contexts and is quickly becoming a popular method to edit primary cells for therapeutic application. This article describes how to make a Cas9 RNP and outlines its use for gene editing in human cells. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

  7. Common Wet Chemical Agents for Purifying Multiwalled Carbon Nanotubes

    Directory of Open Access Journals (Sweden)

    Rasel Das

    2014-01-01

    Full Text Available Purification and functionalization of multiwalled carbon nanotubes (MWCNTs are challenging but vital for their effective applications in various fields including water purification technologies, optoelectronics, biosensors, fuel cells, and electrode arrays. The currently available purification techniques, often complicated and time consuming, yielded shortened and curled MWCNTs that are not suitable for applications in certain fields such as membrane technologies, hybrid catalysis, optoelectronics, and sensor developments. Here we described the H2O2 synergy on the actions of HCl and KOH in purifying and functionalizing pristine MWCNTs. The method (HCl/H2O2 showed 100% purification yield as compared to HCl and KOH/H2O2 with purification yields 93.46 and 3.92%, respectively. We probed the findings using transmission electron microscope, energy dispersive X-ray spectroscope, attenuated total reflectance infrared spectroscope, Raman spectroscope, thermal gravimetric analysis, and X-ray powder diffraction. The study is a new avenue for simple, rapid, low cost, and scalable purification of pristine MWCNTs for application in versatile fields.

  8. Full scale demonstration of air-purifying pavement

    International Nuclear Information System (INIS)

    Ballari, M.M.; Brouwers, H.J.H.

    2013-01-01

    Highlights: ► The results of a demonstration project for photocatalytic pavement are shown. ► The photocatalytic performance was studied in a street as well as on lab scale. ► The outdoor monitoring was performed in different seasons and weather conditions. ► The NO x concentration was in average 19% lowered by the photocatalytic street. ► Under ideal weather conditions the NO x reduction reached up to 45%. -- Abstract: Experiments concerning a full-scale demonstration of air purifying pavement in Hengelo, The Netherlands, are reported. The full width of the street was provided with concrete pavement containing TiO 2 over a length of 150 m (“DeNO x street”). Another part of the street, about 100 m, was paved with normal paving blocks (“Control street”). The outdoor monitoring was done during 26 days for a period exceeding one year, and measured parameters included traffic intensity, NO, NO 2 and ozone concentrations, temperature, relative humidity, wind speed and direction, and the visible and UV light irradiance. Prior and parallel to these field measurements, the used blocks were also measured in the lab to assess their performance. The NO x concentration was, on average, 19% (considering the whole day) and 28% (considering only afternoons) lower than the obtained values in the Control street. Under ideal weather conditions (high radiation and low relative humidity) a NO x concentration decrease of 45% could be observed

  9. Full scale demonstration of air-purifying pavement

    Energy Technology Data Exchange (ETDEWEB)

    Ballari, M.M., E-mail: ballari@santafe-conicet.gov.ar [Department of the Built Environment, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven (Netherlands); Brouwers, H.J.H., E-mail: jos.brouwers@tue.nl [Department of the Built Environment, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven (Netherlands)

    2013-06-15

    Highlights: ► The results of a demonstration project for photocatalytic pavement are shown. ► The photocatalytic performance was studied in a street as well as on lab scale. ► The outdoor monitoring was performed in different seasons and weather conditions. ► The NO{sub x} concentration was in average 19% lowered by the photocatalytic street. ► Under ideal weather conditions the NO{sub x} reduction reached up to 45%. -- Abstract: Experiments concerning a full-scale demonstration of air purifying pavement in Hengelo, The Netherlands, are reported. The full width of the street was provided with concrete pavement containing TiO{sub 2} over a length of 150 m (“DeNO{sub x} street”). Another part of the street, about 100 m, was paved with normal paving blocks (“Control street”). The outdoor monitoring was done during 26 days for a period exceeding one year, and measured parameters included traffic intensity, NO, NO{sub 2} and ozone concentrations, temperature, relative humidity, wind speed and direction, and the visible and UV light irradiance. Prior and parallel to these field measurements, the used blocks were also measured in the lab to assess their performance. The NO{sub x} concentration was, on average, 19% (considering the whole day) and 28% (considering only afternoons) lower than the obtained values in the Control street. Under ideal weather conditions (high radiation and low relative humidity) a NO{sub x} concentration decrease of 45% could be observed.

  10. Chitosanase purified from bacterial isolate Bacillus licheniformis of ruined vegetables displays broad spectrum biofilm inhibition.

    Science.gov (United States)

    Muslim, Sahira Nsayef; Al-Kadmy, Israa M S; Hussein, Nadheema Hammood; Mohammed Ali, Alaa Naseer; Taha, Buthainah Mohammed; Aziz, Sarah Naji; Kheraif, Abdulaziz Abdullah Al; Divakar, Darshan Devang; Ramakrishnaiah, Ravikumar

    2016-11-01

    A number of bacterial species produces chitosanases which has variety of applications because of its high biodegradability, non-toxicity and antimicrobial assets. In the present study chitosanase is purified from new bacterial species Bacillus licheniformis from spoiled vegetable. This novel strain of Bacillus licheniformis isolated from spoilt cucumber and pepper samples has the ability to produce the chitosanase enzyme when grown on chitosan substrate. Study also examined its antibiofilm properties against diverse bacterial species with biofilm forming ability. The purified chitosanase inhibited the biofilm formation ability for all Gram-negative and Gram-positive biofilm-forming bacteria [biofilm producers] tested in this study in congo red agar and microtiter plate's methods. Highly antibiofilm activity of chitosanase was recorded against Pseudomonas aeruginosa followed by Klebsiella pneumoniae with reduction of biofilm formation upto 22 and 29%, respectively compared with [100] % of control. Biofilm formation has multiple role including ability to enhance resistance and self-protection from external stress. This chitosanase has promising benefit as antibiofilm agent against biofilm forming pathogenic bacteria and has promising application as alternative antibiofilm agents to combat the growing number of multidrug resistant pathogen-associated infections, especially in situation where biofilms are involved. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. serological screening for ante-natal toxoplasmosis in maiduguri ...

    African Journals Online (AJOL)

    parasite, Toxoplasma gondii (T. gondii), an obligate intracellular protozoan ... In this study, we examined the immunological status of pregnant .... transplacentally and parasite can cause systemic infection, we posit .... actors for Taenia saginata.

  12. The inhibition of the Human Immunodeficiency Virus type 1 activity by crude and purified human pregnancy plug mucus and mucins in an inhibition assay

    Directory of Open Access Journals (Sweden)

    Schoeman Leann

    2008-05-01

    Full Text Available Abstract Background The female reproductive tract is amongst the main routes for Human Immunodeficiency Virus (HIV transmission. Cervical mucus however is known to protect the female reproductive tract from bacterial invasion and fluid loss and regulates and facilitates sperm transport to the upper reproductive tract. The purpose of this study was to purify and characterize pregnancy plug mucins and determine their anti-HIV-1 activity in an HIV inhibition assay. Methods Pregnancy plug mucins were purified by caesium chloride density-gradient ultra-centrifugation and characterized by Western blotting analysis. The anti-HIV-1 activities of the crude pregnancy plug mucus and purified pregnancy plug mucins was determined by incubating them with HIV-1 prior to infection of the human T lymphoblastoid cell line (CEM SS cells. Results The pregnancy plug mucus had MUC1, MUC2, MUC5AC and MUC5B. The HIV inhibition assay revealed that while the purified pregnancy plug mucins inhibit HIV-1 activity by approximately 97.5%, the crude pregnancy plug mucus failed to inhibit HIV-1 activity. Conclusion Although it is not clear why the crude sample did not inhibit HIV-1 activity, it may be that the amount of mucins in the crude pregnancy plug mucus (which contains water, mucins, lipids, nucleic acids, lactoferrin, lysozyme, immunoglobulins and ions, is insufficient to cause viral inhibition or aggregation.

  13. Morphologic and genetic identification of Taenia tapeworms in Tanzania and DNA genotyping of Taenia solium.

    Science.gov (United States)

    Eom, Keeseon S; Chai, Jong-Yil; Yong, Tai-Soon; Min, Duk-Young; Rim, Han-Jong; Kihamia, Charles; Jeon, Hyeong-Kyu

    2011-12-01

    Species identification of Taenia tapeworms was performed using morphologic observations and multiplex PCR and DNA sequencing of the mitochondrial cox1 gene. In 2008 and 2009, a total of 1,057 fecal samples were collected from residents of Kongwa district of Dodoma region, Tanzania, and examined microscopically for helminth eggs and proglottids. Of these, 4 Taenia egg positive cases were identified, and the eggs were subjected to DNA analysis. Several proglottids of Taenia solium were recovered from 1 of the 4 cases. This established that the species were T. solium (n = 1) and T. saginata (n = 3). One further T. solium specimen was found among 128 fecal samples collected from Mbulu district in Arusha, and this had an intact strobila with the scolex. Phylegenetic analysis of the mtDNA cox1 gene sequences of these 5 isolates showed that T. saginata was basal to the T. solium clade. The mitochondrial cox1 gene sequences of 3 of these Tanzanian isolates showed 99% similarity to T. saginata, and the other 2 isolates showed 100% similarity to T. solium. The present study has shown that Taenia tapeworms are endemic in Kongwa district of Tanzania, as well as in a previously identified Mbulu district. Both T. solium isolates were found to have an "African/Latin American" genotype (cox1).

  14. Effect of preparation conditions on the characteristics and photocatalytic activity of TiO2/purified diatomite composite photocatalysts

    Science.gov (United States)

    Sun, Zhiming; Hu, Zhibo; Yan, Yang; Zheng, Shuilin

    2014-09-01

    TiO2/purified diatomite composite materials were prepared through a modified hydrolysis-deposition method under low temperature using titanium tetrachloride as precursor combined with a calcination crystallization process. The microstructure and crystalline phases of the obtained composites prepared under different preparation conditions were characterized by high resolution scanning electron microscope (SEM) and X-ray diffraction (XRD), respectively. The photocatalytic performance of TiO2/purified diatomite composites was evaluated by Rhodamine B as the target pollutant under UV irradiation, and the optimum preparation conditions of composites were obtained. The TiO2 crystal form in composites prepared under optimum conditions was anatase, the grain size of which was 34.12 nm. The relationships between structure and property of composite materials were analyzed and discussed. It is indicated that the TiO2 nanoparticles uniformly dispersed on the surface of diatoms, and the photocatalytic performance of the composite materials was mainly determined by the dispersity and grain size of loaded TiO2 nanoparticles.

  15. Development of intelligent monitoring purifier for indoor PM 2.5

    Science.gov (United States)

    Lou, Guanting; Zhu, Rong; Guo, Jiangwei; Wei, Yongqing

    2018-03-01

    The particulate matter 2.5 (PM2.5) refers to tiny particles or droplets in the air that are two and one half microns or less in width. PM2.5 is an air pollutant that is a concern for people’s health when levels in air are high. The intelligent monitoring purifier was developed to detect indoor PM2.5 concentration before and after purification and the monitoring data could be displayed on the LCD screen, displaying different color patterns according to the concentrations. Through the Bluetooth transport module, real-time values could also display on the mobile phone and voice broadcast PM2.5 concentration level in the air. When PM2.5 concentration is higher than the setting threshold, the convection fan rotation and the speed can be remote controlled with mobile phone through the Bluetooth transport. Therefore, the efficiency and scope of the purification could be enhanced and further better air quality could be achieved.

  16. Requirements for growth and IL-10 expression of highly purified human T regulatory cells

    Science.gov (United States)

    Bonacci, Benedetta; Edwards, Brandon; Jia, Shuang; Williams, Calvin; Hessner, Martin J.; Gauld, Stephen; Verbsky, James

    2013-01-01

    Human regulatory T cells (TR) cells have potential for the treatment of a variety of immune mediated diseases but the anergic phenotype of these cells makes them difficult to expand in vitro. We have examined the requirements for growth and cytokine expression from highly purified human TR cells, and correlated these findings with the signal transduction events of these cells. We demonstrate that these cells do not proliferate or secrete IL-10 even in the presence of high doses of IL-2. Stimulation with a superagonistic anti-CD28 antibody (clone 9D4) and IL-2 partially reversed the proliferative defect, and this correlated with reversal of the defective calcium mobilization in these cells. Dendritic cells were effective at promoting TR cell proliferation, and under these conditions the proliferative capacity of TR cells was comparable to conventional CD4 lymphocytes. Blocking TGF-β activity abrogated IL-10 expression from these cells, while addition of TGF-β resulted in IL-10 production. These data demonstrate that highly purified populations of TR cells are anergic even in the presence of high doses of IL-2. Furthermore, antigen presenting cells provide proper co-stimulation to overcome the anergic phenotype of TR cells, and under these conditions they are highly sensitive to IL-2. In addition, these data demonstrate for the first time that TGF-β is critical to enable human TR cells to express IL-10. PMID:22562448

  17. Purifying Selection Maintains Dosage-Sensitive Genes during Degeneration of the Threespine Stickleback Y Chromosome

    Science.gov (United States)

    White, Michael A.; Kitano, Jun; Peichel, Catherine L.

    2015-01-01

    Sex chromosomes are subject to unique evolutionary forces that cause suppression of recombination, leading to sequence degeneration and the formation of heteromorphic chromosome pairs (i.e., XY or ZW). Although progress has been made in characterizing the outcomes of these evolutionary processes on vertebrate sex chromosomes, it is still unclear how recombination suppression and sequence divergence typically occur and how gene dosage imbalances are resolved in the heterogametic sex. The threespine stickleback fish (Gasterosteus aculeatus) is a powerful model system to explore vertebrate sex chromosome evolution, as it possesses an XY sex chromosome pair at relatively early stages of differentiation. Using a combination of whole-genome and transcriptome sequencing, we characterized sequence evolution and gene expression across the sex chromosomes. We uncovered two distinct evolutionary strata that correspond with known structural rearrangements on the Y chromosome. In the oldest stratum, only a handful of genes remain, and these genes are under strong purifying selection. By comparing sex-linked gene expression with expression of autosomal orthologs in an outgroup, we show that dosage compensation has not evolved in threespine sticklebacks through upregulation of the X chromosome in males. Instead, in the oldest stratum, the genes that still possess a Y chromosome allele are enriched for genes predicted to be dosage sensitive in mammals and yeast. Our results suggest that dosage imbalances may have been avoided at haploinsufficient genes by retaining function of the Y chromosome allele through strong purifying selection. PMID:25818858

  18. Use of recombinant purified protein derivative (PPD) antigens as specific skin test for tuberculosis.

    Science.gov (United States)

    Stavri, Henriette; Bucurenci, Nadia; Ulea, Irina; Costache, Adriana; Popa, Loredana; Popa, Mircea Ioan

    2012-11-01

    Purified protein derivative (PPD) is currently the only available skin test reagent used worldwide for the diagnosis of tuberculosis (TB). The aim of this study was to develop a Mycobacterium tuberculosis specific skin test reagent, without false positive results due to Bacillus Calmette-Guerin (BCG) vaccination using recombinant antigens. Proteins in PPD IC-65 were analyzed by tandem mass spectrometry and compared to proteins in M. tuberculosis culture filtrate; 54 proteins were found in common. Top candidates MPT64, ESAT 6, and CFP 10 were overexpressed in Escherichia coli expression strains and purified as recombinant proteins. To formulate optimal immunodiagnostic PPD cocktails, the antigens were evaluated by skin testing guinea pigs sensitized with M. tuberculosis H37Rv and BCG. For single antigens and a cocktail mixture of these antigens, best results were obtained using 3 μg/0.1 ml, equivalent to 105 TU (tuberculin units). Each animal was simultaneously tested with PPD IC-65, 2 TU/0.1 ml, as reference. Reactivity of the multi-antigen cocktail was greater than that of any single antigen. The skin test results were between 34.3 and 76.6 per cent the level of reactivity compared to that of the reference when single antigens were tested and 124 per cent the level of reactivity compared to the reference for the multi-antigen cocktail. Our results showed that this specific cocktail could represent a potential candidate for a new skin diagnostic test for TB.

  19. In vivo behavior of detergent-solubilized purified rabbit thrombomodulin on intravenous injection into rabbits

    International Nuclear Information System (INIS)

    Ehrlich, H.J.; Esmon, N.L.; Bang, N.U.

    1990-01-01

    Thrombomodulin is a thrombin endothelial cell membrane receptor. The thrombomodulin-thrombin complex rapidly activates protein C resulting in anticoagulant activity. We investigated the anticoagulant effects and pharmacokinetic behavior of detergent-solubilized purified rabbit thrombomodulin labeled with iodine 125 when intravenously injected into rabbits. Thrombomodulin half-life (t1/2) was determined by tracking the 125I-radiolabeled protein and the biologic activity as determined by the prolongation of the activated partial thromboplastin time (APTT) and thrombin clotting time (TCT). When 200 micrograms/kg 125I-thrombomodulin was injected into rabbits, the APTT and TCT were immediately prolonged, whereas no effect on the prothrombin time was seen. In vitro calibration curves enabled us to convert the prolongations of the clotting times into micrograms per milliliter thrombomodulin equivalents. The best fit (r greater than 0.99) for the disappearance curves was provided by a two-compartment model with mean t1/2 alpha (distribution phase) of 18 minutes for 125I, 12 minutes for APTT, and 20 minutes for TCT, and mean t1/2 beta (elimination phase) of 385 minutes for 125I, 460 for APTT, and 179 for TCT. The administration of two doses of endotoxin (50 micrograms/kg) 24 hours apart did not accelerate the turnover rate of 125I-thrombomodulin as measured by the disappearance of 125I from the circulation. Thus, detergent-solubilized purified thrombomodulin administered intravenously circulates in a biologically active form for appreciable time periods

  20. Method of purifying uranium tetrafluoride hydrate and preparing uranium (VI) peroxide hydrate using a fluoride complexing agent

    International Nuclear Information System (INIS)

    Barreiro, A.J.; Lowe, C.M.T.; Lefever, J.A.; Pyman, R.L.

    1983-01-01

    The annual production of phosphate rock, on the order of about 30-40 million tons yearly, represents several million pounds of uranium. The present invention provides a process of purifying uranium tetrafluoride hydrate to produce a uranium (VI) peroxide product meeting 'yellow cake' standards using a double precipitation procedure. A fluoride complexing agent is used in the precipitation

  1. Cystercercus larva in the anterior chamber of the eye of a 7year o

    African Journals Online (AJOL)

    user

    dogs (T. solium, Echinococcus), cows (T. saginata), rodents. (Hymenolepsis), and fish ... Systemic examination was essentially normal. An assessment of .... David S, Mathai E. Ocular cystercercosis: a review of 25 cases. J Assoc Physicians ...

  2. Influence of Heat Treatment on the Corrosion Behavior of Purified Magnesium and AZ31 Alloy

    OpenAIRE

    Khalifeh, Sohrab; Burleigh, T. David

    2017-01-01

    Magnesium and its alloys are ideal for biodegradable implants due to their biocompatibility and their low-stress shielding. However, they can corrode too rapidly in the biological environment. The objective of this research was to develop heat treatments to slow the corrosion of high purified magnesium and AZ31 alloy in simulated body fluid at 37{\\deg}C. Heat treatments were performed at different temperatures and times. Hydrogen evolution, weight loss, PDP, and EIS methods were used to measu...

  3. Production, characterization, and immobilization of partially purified surfactant-detergent and alkali-thermostable protease from newly isolated Aeromonas caviae.

    Science.gov (United States)

    Datta, Sumitra; Menon, Gopalakrishnan; Varughese, Bincy

    2017-04-21

    Proteolytic Aeromonas caviae P-1-1 growing at wide-ranging pH (7.0-11.0) and moderate salinity (0-5% NaCl) was isolated from cattle shed of Thanjavur, India. It produced lipase, gelatinase, and polyhydroxybutyrate. Different culture conditions, incubation time, carbon and nitrogen sources, vitamins, amino acids, surfactants, and metal ions for optimal growth and protease production of P-1-1 were examined. Maximum protease (0.128 U/mL) production was achieved with 1% fructose, 1% yeast extract, 0.1% ammonium sulfate, 3% NaCl, 0.1% CaCl 2  · 2H 2 O, 1% glycine, 0.1% vitamin E, and 0.1% Tween-40 at pH 8.0 after 42 hr of incubation at 37°C. It was active over broad range of pH (7.0-12.0), temperature (15-100°C), and salinity (0-9% NaCl) with optima at pH 10.0, 55°C, and 3% NaCl. It retained 65 and 48% activities at pH 12.0 and 100°C, respectively. Partially purified protease was highly stable (100%) within pH range 7.0-12.0 and salinities of 0-5% NaCl for 48 hr. Cu 2+ , Mn 2+ , Co 2+ , and Ca 2+ did not inhibit its activity. Its stability at extreme pHs, temperatures, and in the presence of surfactants and commercial detergents suggests its possible application in laundry detergents. Partially purified protease was immobilized and reused. This is the first report of alkali-thermotolerant, surfactant-detergent-stable partially purified extracellular protease from A. caviae.

  4. Naloxone-sensitive, haloperidol-sensitive, [3H](+)SKF-10047-binding protein partially purified from rat liver and rat brain membranes: an opioid/sigma receptor?

    Science.gov (United States)

    Tsao, L I; Su, T P

    1997-02-01

    A naloxone-sensitive, haloperidol-sensitive, [3H](+)SKF-10047-binding protein was partially purified from rat liver and rat brain membranes in an affinity chromatography originally designed to purify sigma receptors. Detergent-solubilized extracts from membranes were adsorbed to Sephadex G-25 resin containing an affinity ligand for sigma receptors: N-(2- 3,4-dichlorophenyl]ethyl)-N-(6-aminohexyl)-(2-[1-pyrrolidinyl]) ethylamine (DAPE). After eluting the resin with haloperidol, a protein that bound [3H](+)SKF-10047 was detected in the eluates. However, the protein was not the sigma receptor. [3H](+)SKF-10047 binding to the protein was inhibited by the following compounds in the order of decreasing potency: (+)pentazocine > (-) pentazocine > (+/-)cyclazocine > (-)morphine > (-)naloxone > haloperidol > (+)SKF-10047 > DADLE > (-)SKF-10047. Further, the prototypic sigma receptor ligands, such as 1,3-di-o-tolylguanidine (DTG), (+)3-PPP, and progesterone, bound poorly to the protein. Tryptic digestion and heat treatment of the affinity-purified protein abolished radioligand binding. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE) of the partially-purified protein from the liver revealed a major diffuse band with a molecular mass of 31 kDa, a polypeptide of 65 kDa, and another polypeptide of > 97 kDa. This study demonstrates the existence of a novel protein in the rat liver and rat brain which binds opioids, benzomorphans, and haloperidol with namomolar affinity. The protein resembles the opioid/sigma receptor originally proposed by Martin et al. [(1976): J. Pharmacol. Exp. Ther., 197:517-532.]. A high degree of purification of this protein has been achieved in the present study.

  5. Workplace performance of a loose-fitting powered air purifying respirator during nanoparticle synthesis

    International Nuclear Information System (INIS)

    Koivisto, Antti J.; Aromaa, Mikko; Koponen, Ismo K.; Fransman, Wouter; Jensen, Keld A.; Mäkelä, Jyrki M.; Hämeri, Kaarle J.

    2015-01-01

    Nanoparticle (particles with diameter ≤100 nm) exposure is recognized as a potentially harmful size fraction for pulmonary particle exposure. During nanoparticle synthesis, the number concentrations in the process room may exceed 10 × 10 6 cm −3 . During such conditions, it is essential that the occupants in the room wear highly reliable high-performance respirators to prevent inhalation exposure. Here we have studied the in-use program protection factor (PPF) of loose-fitting powered air purifying respirators, while workers were coating components with TiO 2 or Cu x O y nanoparticles under a hood using a liquid flame spray process. The PPF was measured using condensation particle counters, an electrical low pressure impactor, and diffusion chargers. The room particle concentrations varied from 4 × 10 6 to 40 × 10 6 cm −3 , and the count median aerodynamic diameter ranged from 32 to 180 nm. Concentrations inside the respirator varied from 0.7 to 7.2 cm −3 . However, on average, tidal breathing was assumed to increase the respirator concentration by 2.3 cm −3 . The derived PPF exceeded 1.1 × 10 6 , which is more than 40 × 10 3 times the respirator assigned protection factor. We were unable to measure clear differences in the PPF of respirators with old and new filters, among two male and one female user, or assess most penetrating particle size. This study shows that the loose-fitting powered air purifying respirator provides very efficient protection against nanoparticle inhalation exposure if used properly

  6. Approaches to raw sugar quality improvement as a route to sustaining a reliable supply of purified industrial sugar feedstocks

    Science.gov (United States)

    Energy costs in the sugar industry are outstripping costs of manufacture, particularly in refineries. This, as well as increasing transportation costs and the need to meet manufacturers’ tight specifications, has increased the demand for a sustainable supply of purified, raw sugar. Agricultural com...

  7. Some properties of purified hepatoredoxin from bovine liver mitochondria

    International Nuclear Information System (INIS)

    Gilevich, S.N.; Gurev, O.L.; Shkumatov, V.M.; Chashchin, V.L.; Akhrem, A.A.

    1986-01-01

    Some of the most important physicochemical properties of hepatoredoxin from bovine liver, purified to a homogeneous state, were determined for the first time. The protein contains a [2Fe-2S] cluster in its active site and in an oxidized state has absorption maxima at 280, 320, 415, and 455 nm. The spectrophotometric index of purity (A 415 /A 280 ) of the homogeneous native preparation is 0.84; the extinction coefficient (epsilon 415 ) is equal to 9800 M -1 cm -1 . According to the data of gel electrophoresis in the presence of SDS, hepatoredoxin has an M/sub r/ of 12,500; its isoelectric point (pI) is equal to 4.2. Hepatoredoxin is necessary for the reconstitution of the C 27 -steroid hydroxylase activity and can be replaced by the related protein, adrenodoxin. All the parameters listed above, as well as the CD spectra, the immunochemical properties, and sequence of the first five N-terminal amino acids of hepatoredoxin and adrenodoxin are very similar of identical. At the same time, the amino acid composition of the two ferredoxins, along with common properties, has some differences

  8. Process for recovering a uranium containing concentrate and purified phosphoric acid from a wet process phosphoric acid containing uranium

    International Nuclear Information System (INIS)

    Weterings, C.A.M.; Janssen, J.A.

    1985-01-01

    A process is claimed for recovering from a wet process phosphoric acid which contains uranium, a uranium containing concentrate and a purified phosphoric acid. The wet process phosphoric acid is treated with a precipitant in the presence of a reducing agent and an aliphatic ketone

  9. Process for recovering a uranium containing concentrate and purified phosphoric acid from a wet process phosphoric acid containing uranium

    Energy Technology Data Exchange (ETDEWEB)

    Weterings, C.A.M.; Janssen, J.A.

    1985-04-30

    A process is claimed for recovering from a wet process phosphoric acid which contains uranium, a uranium containing concentrate and a purified phosphoric acid. The wet process phosphoric acid is treated with a precipitant in the presence of a reducing agent and an aliphatic ketone.

  10. Development of DNA affinity techniques for the functional characterization of purified RNA polymerase II transcription factors

    International Nuclear Information System (INIS)

    Garfinkel, S.; Thompson, J.A.; Cohen, R.B.; Brendler, T.; Safer, B.

    1987-01-01

    Affinity adsorption, precipitation, and partitioning techniques have been developed to purify and characterize RNA Pol II transcription components from whole cell extracts (WCE) (HeLa) and nuclear extracts (K562). The titration of these extracts with multicopy constructs of the Ad2 MLP but not pUC8, inhibits transcriptional activity. DNA-binding factors precipitated by this technique are greatly enriched by centrifugation. Using this approach, factors binding to the upstream promoter sequence (UPS) of the Ad2 MLP have been rapidly isolated by Mono Q, Mono S, and DNA affinity chromatography. By U.V. crosslinking to nucleotides containing specific 32 P-phosphodiester bonds within the recognition sequence, this factor is identified as a M/sub r/ = 45,000 polypeptide. To generate an assay system for the functional evaluation of single transcription components, a similar approach using synthetic oligonucleotide sequences spanning single promoter binding sites has been developed. The addition of a synthetic 63-mer containing the UPS element of the Ad2 MLP to HeLa WCE inhibited transcription by 60%. The addition of partially purified UPS binding protein, but not RNA Pol II, restored transcriptional activity. The addition of synthetic oligonucleotides containing other regulatory sequences not present in the Ad2 MLP was without effect

  11. The fate of purified radio-labelled alkaline phosphatase from the liver in the organism

    International Nuclear Information System (INIS)

    Herbert, V.

    1981-01-01

    Alkaline phosphatase (AP) from dog liver was enriched by a factor of 5.444 in various steps. Rabbit antiserum to the purified AP was produced; 125-I was used then to radiolabel the highly purified AP. Four dogs were cholecystectomized and subsequently received an extracorporal drainage of the bile ducts. Decrease rate of total radio-activity and of PBI in the serum was determined in one dog; likewise in three other dogs before and one week after occlusion of their main bile ducts. In addition, radioactivity above the organs was measured in some animals at short intervals. In the dogs with main bile duct drainage, bile was collected continuously for up to 70 h, samples were taken, and residual bile plus native dog bile were re-infused into the distal choledochus catheter. Total radioactivity, PBI and immunoprecipitability with antibodies were determined in the bile and serum samples. AP, GOT, CPT and bilirubin were determined in some serum samples. In addition, total radioactivity excreted by urine was established. Results show injected 125-I-AP to be rapidly stored in the liver and not to be excreted via bile to a decisive extent. The fact that 125-I-AP is not excreted via bile is further indicated by the identical decrease rate of injected 125-I-AP in the serum in dogs with and without main bile duct occlusion. Injected 125-I-AP appears to be metabolized very rapidly in the liver as is indicated by the rapid decrease of immuno precipitability of 125-I-AP in the serum. (orig./MG) [de

  12. Development of a water purifier for radioactive cesium removal from contaminated natural water by radiation-induced graft polymerization

    Science.gov (United States)

    Seko, Noriaki; Hoshina, Hiroyuki; Kasai, Noboru; Shibata, Takuya; Saiki, Seiichi; Ueki, Yuji

    2018-02-01

    Six years after the Fukushima-nuclear accident, the dissolved radioactive cesium (Cs) is now hardly detected in environmental natural waters. These natural waters are directly used as source of drinking and domestic waters in disaster-stricken areas in Fukushima. However, the possibility that some radioactive Cs adsorbed on soil or leaves will contaminate these natural waters during heavy rains or typhoon is always present. In order for the returning residents to live with peace of mind, it is important to demonstrate the safety of the domestic waters that they will use for their daily life. For this purpose, we have synthesized a material for selective removal of radioactive Cs by introducing ammonium 12-molybdophosphate (AMP) onto polyethylene nonwoven fabric through radiation-induced emulsion graft polymerization technique. Water purifiers filled with the grafted Cs adsorbent were installed in selected houses in Fukushima. The capability of the grafted adsorbent to remove Cs from domestic waters was evaluated for a whole year. The results showed that the tap water filtered through the developed water purifier contained no radioactive Cs, signifying the very effective adsorption performance of the developed grafted adsorbent. From several demonstrations, we have commercialized the water purifier named "KranCsair®". Furthermore, we have also developed a method for the mass production of the grafted nonwoven fabric. Using a 30 L grafting reactor, it was possible to produce the grafted nonwoven fabric with a suitable range of degree of grafting. When an irradiated roll of nonwoven trunk fabric with a length of 10 m and a width of 30 cm was set in the reactor filled with glycidyl methacrylate (GMA), AMP, Tween 80 monomer emulsion solution at 40 °C for 1 h, the difference of Dgs in the length and the width on roll of fabrics was negligible.

  13. A Rapid and Efficient Method for Purifying High Quality Total RNA from Peaches (Prunus persica for Functional Genomics Analyses

    Directory of Open Access Journals (Sweden)

    LEE MEISEL

    2005-01-01

    Full Text Available Prunus persica has been proposed as a genomic model for deciduous trees and the Rosaceae family. Optimized protocols for RNA isolation are necessary to further advance studies in this model species such that functional genomics analyses may be performed. Here we present an optimized protocol to rapidly and efficiently purify high quality total RNA from peach fruits (Prunus persica. Isolating high-quality RNA from fruit tissue is often difficult due to large quantities of polysaccharides and polyphenolic compounds that accumulate in this tissue and co-purify with the RNA. Here we demonstrate that a modified version of the method used to isolate RNA from pine trees and the woody plant Cinnamomun tenuipilum is ideal for isolating high quality RNA from the fruits of Prunus persica. This RNA may be used for many functional genomic based experiments such as RT-PCR and the construction of large-insert cDNA libraries.

  14. Mixed protocols: Multiple ratios of FSH and LH bioactivity using highly purified, human-derived FSH (BRAVELLE and highly purified hMG (MENOPUR are unaltered by mixing together in the same syringe

    Directory of Open Access Journals (Sweden)

    Raike Elizabeth

    2005-11-01

    Full Text Available Abstract Background The use of mixed or blended protocols, that utilize both FSH and hMG, for controlled ovarian hyperstimulation is increasing in use. To reduce the number of injections a patient must administer, many physicians instruct their patients to mix their FSH and hMG together to be given as a single injection. Therefore, the goal of this study was to definitively determine if the FSH and LH bioactivities of highly purified, human-derived FSH (Bravelle(R and highly purified hMG (Menopur(R were altered by reconstituting in 0.9% saline and mixing in the same syringe. Methods Bravelle(R and Menopur(R were reconstituted in 0.9% saline and mixed in a Becton Dickinson plastic syringe. The FSH and LH bioactivities of the products were determined after injecting female and male rats, respectively, with Bravelle(R, Menopur(R, or a mixture of Bravelle(R and Menopur(R. Ratios of FSH:LH activity tested were 150:75 IU (1 vial Bravelle(R: 1 vial Menopur(R, 300:75 IU (3 vials Bravelle(R: 1 vial Menopur(R or 300:225 IU (1 vial Bravelle(R: 3 vials of Menopur(R. Results There were no statistically significant changes in either FSH or LH bioactivity that occurred after mixing Bravelle(R with Menopur(R in the same syringe. The theoretical vs. actual FSH bioactivity for Bravelle(R and Menopur(R were 75 vs. 76.58 IU/mL and 75 vs. 76.0 IU/mL, respectively. For the 3 ratios of FSH:LH activity tested, 150:75 IU (1 vial Bravelle(R: 1 vial Menopur(R, 300:75 IU (3 vials Bravelle(R: 1 vial Menopur(R or 300:225 IU (1 vial Bravelle(R: 3 vials of Menopur(R tested, the theoretical vs. actual FSH bioactivities were 150 vs. 156.86 IU/mL, 300 vs. 308.69 IU/mL and 300 vs. 306.58 IU/mL, respectively. The theoretical vs. actual LH bioactivity for Menopur(R in the above mentioned ratios tested were 75 vs. 77.50 IU/mL. For the 3 ratios of FSH:LH activity tested, 150:75 IU (1 vial Bravelle(R: 1 vial Menopur(R, 300:75 IU (3 vials Bravelle(R: 1 vial Menopur(R or 300:225 IU (1

  15. Dietary n-3 PUFA affect TcR-mediated activation of purified murine T cells and accessory cell function in co-cultures

    Science.gov (United States)

    CHAPKIN, R S; ARRINGTON, J L; APANASOVICH, T V; CARROLL, R J; MCMURRAY, D N

    2002-01-01

    Diets enriched in n-3 polyunsaturated fatty acids (PUFA) suppress several functions of murine splenic T cells by acting directly on the T cells and/or indirectly on accessory cells. In this study, the relative contribution of highly purified populations of the two cell types to the dietary suppression of T cell function was examined. Mice were fed diets containing different levels of n-3 PUFA; safflower oil (SAF; control containing no n-3 PUFA), fish oil (FO) at 2% and 4%, or 1% purified docosahexaenoic acid (DHA) for 2 weeks. Purified (>90%) T cells were obtained from the spleen, and accessory cells (>95% adherent, esterase-positive) were obtained by peritoneal lavage. Purified T cells or accessory cells from each diet group were co-cultured with the alternative cell type from every other diet group, yielding a total of 16 different co-culture combinations. The T cells were stimulated with either concanavalin A (ConA) or antibodies to the T cell receptor (TcR)/CD3 complex and the costimulatory molecule CD28 (αCD3/αCD28), and proliferation was measured after four days. Suppression of T cell proliferation in the co-cultures was dependent upon the dose of dietary n-3 PUFA fed to mice from which the T cells were derived, irrespective of the dietary treatment of accessory cell donors. The greatest dietary effect was seen in mice consuming the DHA diet (P = 0·034 in the anova; P = 0·0053 in the Trend Test), and was observed with direct stimulation of the T cell receptor and CD28 costimulatory ligand, but not with ConA. A significant dietary effect was also contributed accessory cells (P = 0·033 in the Trend Test). We conclude that dietary n-3 PUFA affect TcR-mediated by T cell activation by both direct and indirect (accessory cell) mechanisms. PMID:12296847

  16. Workplace performance of a loose-fitting powered air purifying respirator during nanoparticle synthesis

    Energy Technology Data Exchange (ETDEWEB)

    Koivisto, Antti J., E-mail: jok@nrcwe.dk [National Research Centre for the Working Environment (Denmark); Aromaa, Mikko [Tampere University of Technology, Department of Physics (Finland); Koponen, Ismo K. [National Research Centre for the Working Environment (Denmark); Fransman, Wouter [TNO (Netherlands); Jensen, Keld A. [National Research Centre for the Working Environment (Denmark); Mäkelä, Jyrki M. [Tampere University of Technology, Department of Physics (Finland); Hämeri, Kaarle J. [University of Helsinki, Department of Physics (Finland)

    2015-04-15

    Nanoparticle (particles with diameter ≤100 nm) exposure is recognized as a potentially harmful size fraction for pulmonary particle exposure. During nanoparticle synthesis, the number concentrations in the process room may exceed 10 × 10{sup 6} cm{sup −3}. During such conditions, it is essential that the occupants in the room wear highly reliable high-performance respirators to prevent inhalation exposure. Here we have studied the in-use program protection factor (PPF) of loose-fitting powered air purifying respirators, while workers were coating components with TiO{sub 2} or Cu{sub x}O{sub y} nanoparticles under a hood using a liquid flame spray process. The PPF was measured using condensation particle counters, an electrical low pressure impactor, and diffusion chargers. The room particle concentrations varied from 4 × 10{sup 6} to 40 × 10{sup 6} cm{sup −3}, and the count median aerodynamic diameter ranged from 32 to 180 nm. Concentrations inside the respirator varied from 0.7 to 7.2 cm{sup −3}. However, on average, tidal breathing was assumed to increase the respirator concentration by 2.3 cm{sup −3}. The derived PPF exceeded 1.1 × 10{sup 6}, which is more than 40 × 10{sup 3} times the respirator assigned protection factor. We were unable to measure clear differences in the PPF of respirators with old and new filters, among two male and one female user, or assess most penetrating particle size. This study shows that the loose-fitting powered air purifying respirator provides very efficient protection against nanoparticle inhalation exposure if used properly.

  17. Producing Biosurfactants from Purified Microorganisms Obtained from Oil-contaminated Soil

    Directory of Open Access Journals (Sweden)

    Nader Mokhtarian

    2010-09-01

    Full Text Available Contamination of soil by crude oil can pose serious problems to ecosystems. Soil washing by solutions containing biosurfactants is one of the most efficient methods for the remediation of contaminated soil by crude oil because it removes not only the crude oil but also heavy metals. In this study, five soil samples were taken from fields exposed to oil compounds over the years in order to produce biosurfactants from microorganisms that were capable of degrading oil compounds. Sixteen such microorganisms were isolated. After cultivation, their emulsification strength was examined using E24 test. From among the experimental microorganisms, a gram-negative and rod-shape microorganism called A-12 showed the greatest value of the E24 test index (36%. For each liter of the culture medium containing 365 mg of microorganisms, 3 gr of the biosurfactant compound was produced and separated as dried powder. The purified biosurfactant was used in the soil washing process. Also, the insulated microorganisms were capable of degrading crude oil floating on wastewaters.

  18. The stability of human, bovine and avian tuberculin purified protein derivative (PPD).

    Science.gov (United States)

    Maes, Mailis; Giménez, José Francisco; D'Alessandro, Adriana; De Waard, Jacobus H

    2011-11-15

    Guidelines recommend storing tuberculin purified protein derivative (PPD) refrigerated. However, especially in developing countries, maintaining the product refrigerated under field conditions can be difficult, limiting its use. Here we determine the effect of prolonged exposure to high temperatures on the potency of human, bovine and avian tuberculin PPD. Human, bovine and avian tuberculin PPD were stored for several weeks exposed to temperatures ranging from 37º to 100ºC. The potency was evaluated in vivo, in sensitized or naturally infected animals. Most test situations didn't affect the biological activity of the tuberculin PPDs and only very long and extreme incubations (several days at 100 °C) compromised the potency. Tuberculin PPD is very stable and can be stored or transported for long periods without refrigeration. 

  19. Hydrolysis of triolein in phospholipid vesicles and microemulsions by a purified rat liver acid lipase.

    Science.gov (United States)

    Burrier, R E; Brecher, P

    1983-10-10

    An acid lipase was purified from rat liver lysosomes. Lipase purification involved affinity chromatography, gel filtration, and stabilization of the purified preparation using ethylene glycol and Triton X-100. A molecular weight of 67,000-69,000 was determined independently using density gradient centrifugation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and gel filtration. To study enzyme action, model substrates were prepared by incorporating radiolabeled triolein into either unilamellar vesicles or microemulsions. Substrates were prepared by cosonicating aqueous dispersions of lecithin and triolein. Formation of vesicles or emulsions depended on the relative amount of each lipid and on sonication conditions. Vesicles were prepared at molar ratios between 70:1 and 26:1 (lecithin:triolein) and the microemulsion preparation at a molar ratio of 1:1. The substrate particles were of similar size (220-250 A) as determined by Bio-Gel A-15m chromatography. Hydrolysis of triolein contained in vesicles or emulsions was similar with respect to pH, temperature, and reaction products. Kinetic studies on vesicles with increasing triolein content showed progressively greater Vmax values (0-0.6 mumol/min/mg), and Vmax for the emulsion was 3.1 mumol/min/mg. Addition of human very low or low density lipoprotein produced a dose-dependent inhibition with both substrates. The results show that synthetically prepared microemulsions are stable and effective substrates for the acid lipase and indicate that surface-oriented triolein is hydrolyzed in both preparations.

  20. THERAPEUTIC EFFECTS OF HIGHLY PURIFIED DE-GLYCOSYLATED GCMAF IN THE IMMUNOTHERAPY OF PATIENTS WITH CHRONIC DISEASES

    OpenAIRE

    Lynda Thyer; Emma Ward; Rodney Smith; Jacopo J.V. Branca; Gabriele Morucci; Massimo Gulisano; David Noakes; Stefania Pacini

    2013-01-01

    The de-Glycosylated vitamin D binding protein is a powerful Macrophage Activating Factor (GcMAF) that shows multiple biological effects that could be exploited in the immunotherapy of tumours, viral infections and autism. Here we report the observation of a series of clinical cases describing the results obtained administering highly purified GcMAF to patients with diverse types of chronic diseases. These are heterogeneous and refer to patients with different types of diseases at different s...

  1. Analysis of cavitation effect for water purifier using electrolysis

    Science.gov (United States)

    Shin, Dong Ho; Ko, Han Seo; Lee, Seung Ho

    2015-11-01

    Water is a limited and vital resource, so it should not be wasted by pollution. A development of new water purification technology is urgent nowadays since the original and biological treatments are not sufficient. The microbubble-aided method was investigated for removal of algal in this study since it overcomes demerits of the existing purification technologies. Thus, the cavitation effect in a venturi-type tube using the electrolysis was analyzed. Ruthenium-coated titanium plates were used as electrodes. Optimum electrode interval and applied power were determined for the electrolysis. Then, the optimized electrodes were installed in the venturi-type tube for generating cavitation. The cavitation effect could be enhanced without any byproduct by the bubbly flow induced by the electrolysis. The optimum mass flow rate and current were determined for the cavitation with the electrolysis. Finally, the visualization techniques were used to count the cell number of algal and microbubbles for the confirmation of the performance. As a result, the energy saving and high efficient water purifier was fabricated in this study. This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Korean government (MEST) (No. 2013R1A2A2A01068653).

  2. Human neurocysticercosis case and an endemic focus of Taenia solium in Lao PDR.

    Science.gov (United States)

    Jeon, Hyeong-Kyu; Yong, Tai-Soon; Sohn, Woon-Mok; Chai, Jong-Yil; Min, Duk-Young; Rim, Han-Jong; Insisiengmay, Bounnaloth; Eom, Keeseon S

    2013-10-01

    A male patient with neurocysticercosis was identified in Montai Village, Xay District, Oudomxay Province, Lao PDR in February 2004. He had a history of diagnosis for neurocysticercosis by a CT scan in Thailand after an onset of epileptic seizure in 1993. A pig in the same district was found to contain Taenia solium metacestodes (=cysticerci); the slaughtered pig body contained more than 2,000 cysticerci. In addition to morphological identification, molecular identification was also performed on the cysticerci by DNA sequencing analysis of the mitochondrial cox1 gene; they were confirmed as T. solium metacestodes. The patient is regarded as an indigenous case of neurocysticercosis infected in an endemic focus of T. solium taeniasis/cysticercosis in Oudomxay Province, Lao PDR.

  3. A Method of Effective Quarry Water Purifying Using Artificial Filtering Arrays

    Science.gov (United States)

    Tyulenev, M.; Garina, E.; Khoreshok, A.; Litvin, O.; Litvin, Y.; Maliukhina, E.

    2017-01-01

    The development of open pit mining in the large coal basins of Russia and other countries increases their negative impact on the environment. Along with the damage of land and air pollution by dust and combustion gases of blasting, coal pits have a significant negative impact on water resources. Polluted quarry water worsens the ecological situation on a much larger area than covered by air pollution and land damage. This significantly worsens the conditions of people living in cities and towns located near the coal pits, and complicates the subsequent restoration of the environment, irreversibly destroying the nature. Therefore, the research of quarry wastewater purifying is becoming an important mater for scholars of technical colleges and universities in the regions with developing open-pit mining. This paper describes the method of determining the basic parameters of the artificial filtering arrays formed on coal pits of Kuzbass (Western Siberia, Russia), and gives recommendations on its application.

  4. Immunomodulatory Activity of Ganoderma atrum Polysaccharide on Purified T Lymphocytes through Ca2+/CaN and Mitogen-Activated Protein Kinase Pathway Based on RNA Sequencing.

    Science.gov (United States)

    Xiang, Quan-Dan; Yu, Qiang; Wang, Hui; Zhao, Ming-Ming; Liu, Shi-Yu; Nie, Shao-Ping; Xie, Ming-Yong

    2017-07-05

    Our previous study has demonstrated that Ganoderma atrum polysaccharide (PSG-1) has immunomodulatory activity on spleen lymphocytes. However, how PSG-1 exerts its effect on purified lymphocytes is still obscure. Thus, this study aimed to investigate the immunomodulatory activity of PSG-1 on purified T lymphocytes and further elucidate the underlying mechanism based on RNA sequencing (RNA-seq). Our results showed that PSG-1 promoted T lymphocytes proliferation and increased the production of IL-2, IFN-γ, and IL-12. Meanwhile, RNA-seq analysis found 394 differentially expressed genes. KEGG pathway analysis identified 20 significant canonical pathways and seven biological functions. Furthermore, PSG-1 elevated intracellular Ca 2+ concentration and calcineurin (CaN) activity and raised the p-ERK, p-JNK, and p-p38 expression levels. T lymphocytes proliferation and the production of IL-2, IFN-γ, and IL-12 were decreased by the inhibitors of calcium channel and mitogen-activated protein kinases (MAPKs). These results indicated that PSG-1 possesses immunomodulatory activity on purified T lymphocytes, in which Ca 2+ /CaN and MAPK pathways play essential roles.

  5. Molecular characterization of Cysticercus tenuicollis of slaughtered livestock in Upper Egypt governorates

    Directory of Open Access Journals (Sweden)

    Mosaab Adl Eldin Omar

    2016-08-01

    Conclusions: Clearly molecular diagnosis for C. tenuicollis infection significantly helps to differentiate it from such other metacestodes as hydatidosis, which manifests a completely different pathogenicity and requires different control programs.

  6. Purified natural pig immunoglobulins can substitute dietary zinc in reducing piglet post weaning diarrhoea

    DEFF Research Database (Denmark)

    Hedegaard, Chris Juul; Lauridsen, Charlotte; Heegaard, Peter M. H.

    2017-01-01

    antibiotic resistance and pose environmental problems. Recently, in an experimental model of PWD, we observed that oral administration of purified porcine immunoglobulin G (ppIgG) from pooled natural pig plasma could reduce enteric infection. In the present study we were able to reproduce these results...... as it was observed that oral ppIgG accelerated clearance of faecal haemolytic bacteria in pigs challenged with E. coli in comparison with pigs not receiving ppIgG. This effect was observed upon feeding ppIgG for seven days postweaning suggesting that ppIgG does not have to be used prophylactically for several days...

  7. Highly purified HMG versus recombinant FSH for ovarian stimulation in IVF cycles

    DEFF Research Database (Denmark)

    Platteau, P.; Nyboe, Andersen A.; Loft, A.

    2008-01-01

    The objective of this study was to compare the live birth rates resulting from ovarian stimulation with highly purified human menopausal gonadotrophin (HP-HMG), which combines FSH and human chorionic gonadotrophin-driven LH activities, or recombinant FSH (rFSH) alone in women undergoing IVF cycles....... An integrated analysis was performed of the raw data from two randomized controlled trials that were highly comparable in terms of eligibility criteria and post-randomization treatment regimens with either HP-HMG or rFSH for ovarian stimulation in IVF, following a long down-regulation protocol. All randomized...... subjects who received at least one dose of gonadotrophin in an IVF cycle (HP-HMG, n = 491; rFSH, n = 495) were included in the analysis. Subjects who underwent intracytoplasmic sperm injection cycles were excluded. The superiority of one gonadotrophin preparation over the other was tested using...

  8. Cation gating and selectivity in a purified, reconstituted, voltage-dependent sodium channel

    International Nuclear Information System (INIS)

    Barchi, R.L.; Tanaka, J.C.

    1984-01-01

    In excitable membranes, the voltage-dependent sodium channel controls the primary membrane conductance change necessary for the generation of an action potential. Over the past four decades, the time- and voltage-dependent sodium currents gated by this channel have been thoroughly documented with increasingly sophisticated voltage-clamp techniques. Recent advances in the biochemistry of membrane proteins have led to the solubilization and purification of this channel protein from nerve (6) and from muscle (4) or muscle-derived (1) membranes, and have provided an approach to the correlation of the channel's molecular structure with its functional properties. Each of these sodium channel preparations appears to contain a large glycoprotein either as its sole component (2) or in association with several small subunits (6, 3). Evidence that these purified proteins represent the excitable membrane sodium channel is presented. 8 refs., 1 fig., 1 tab

  9. Characterization of the cytochrome c oxidase in isolated and purified plasma membranes from the cyanobacterium Anacystis nidulans

    International Nuclear Information System (INIS)

    Peschek, G.A.; Wastyn, M.; Trnka, M.; Molitor, V.; Fry, I.V.; Packer, L.

    1989-01-01

    Functionally intact plasma membranes were isolated from the cyanobacterium (blue-green alga) Anacystis nidulans through French pressure cell extrusion of lysozyme/EDTA-treated cells, separated from thylakoid membranes by discontinuous sucrose density gradient centrifugation, and purified by repeated recentrifugation. Origin and identity of the chlorophyll-free plasma membrane fraction were confirmed by labeling of intact cells with impermeant protein markers, [ 35 S]diazobenzenesulfonate and fluorescamine, prior to membrane isolation. Rates of oxidation of reduced horse heart cytochrome c by purified plasma and thylakoid membranes were 90 and 2 nmol min -1 (mg of protein) -1 , respectively. The cytochrome oxidase in isolated plasma membranes was identified as a copper-containing aa 3 -type enzyme from the properties of its redox-active and EDTA-resistant Cu 2+ ESR signal, the characteristic inhibition profile, reduced minus oxidized difference spectra, carbon monoxide difference spectra, photoaction and photodissociation spectra of the CO-inhibited enzyme, and immunological cross-reaction of two subunits of the enzyme with antibodies against subunits I and II, and the holoenzyme, of Paracoccus denitrificans aa 3 -type cytochrome oxidase. The data presented are the first comprehensive evidence for the occurrence of aa 3 -type cytochrome oxidase in the plasma membrane of a cyanobacterium similar to the corresponding mitochondrial enzyme

  10. Antioxidant and antibacterial activities of polysaccharides isolated and purified from Diaphragma juglandis fructus.

    Science.gov (United States)

    Meng, Qingran; Li, Yinghao; Xiao, Tiancun; Zhang, Lianfu; Xu, Dan

    2017-12-01

    A water-soluble polysaccharide fraction (DJP-2) isolated from Diaphragma juglandis was successfully purified by ion-exchange chromatography (DEAE-cellulose) and gel-permeation chromatography (Sephadex G-100). The weight-average molecular weight (Mw) and number-average molecular weight (Mn) of DJP-2 were 4.95 and 3.99kDa, respectively. Monosaccharide component analysis indicated that DJP-2 comprised arabinose, galactose, glucose, xylose, and mannose in a molar ratio of 0.27:0.55:1:0.14:0.08. The evaluation of the antioxidant and antibacterial activities of polysaccharides from Diaphragma juglandis fructus indicated that they could be explored as promising natural antioxidant and bacteriostatic agents in the food and pharmaceutical industries. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Subcutaneous infection model facilitates treatment assessment of secondary Alveolar echinococcosis in mice.

    Directory of Open Access Journals (Sweden)

    Tatiana Küster

    Full Text Available Alveolar echinococcosis (AE in humans is a parasitic disease characterized by severe damage to the liver and occasionally other organs. AE is caused by infection with the metacestode (larval stage of the fox tapeworm Echinococcus multilocularis, usually infecting small rodents as natural intermediate hosts. Conventionally, human AE is chemotherapeutically treated with mebendazole or albendazole. There is, however still the need for improved chemotherapeutical options. Primary in vivo studies on drugs of interest are commonly performed in small laboratory animals such as mice and Mongolian jirds, and in most cases, a secondary infection model is used, whereby E. multilocularis metacestodes are directly injected into the peritoneal cavity or into the liver. Disadvantages of this methodological approach include risk of injury to organs during the inoculation and, most notably, a limitation in the macroscopic (visible assessment of treatment efficacy. Thus, in order to monitor the efficacy of chemotherapeutical treatment, animals have to be euthanized and the parasite tissue dissected. In the present study, mice were infected with E. multilocularis metacestodes through the subcutaneous route and were then subjected to chemotherapy employing albendazole. Serological responses to infection were comparatively assessed in mice infected by the conventional intraperitoneal route. We demonstrate that the subcutaneous infection model for secondary AE facilitates the assessment of the progress of infection and drug treatment in the live animal.

  12. A sylvatic lifecycle of Echinococcus equinus in the Etosha National Park, Namibia

    Science.gov (United States)

    Wassermann, Marion; Aschenborn, Ortwin; Aschenborn, Julia; Mackenstedt, Ute; Romig, Thomas

    2014-01-01

    Various species of Echinococcus have been described in the past from wild mammals of sub-Saharan Africa. However, it is only recently, that a few isolates have become available for molecular identification; therefore, the involvement of wildlife in the lifecycles of the various cryptic species within Echinococcus granulosus sensu lato is still only partially known. A preliminary survey was undertaken in Etosha National Park, Namibia, from August to October 2012. Faecal samples were obtained from 34 individual wild carnivores, and metacestodes were collected from carcasses of 18 culled herbivores. Single eggs and metacestode tissue were lysed and identified from sequences of the mitochondrial nad1 gene. In case of metacestodes, the cox1 gene was additionally sequenced and haplotype networks were constructed. Echinococcus equinus was found in lions (4 of 6), black-backed jackals (2 of 7) and Burchell's zebras (11 of 12). The frequency of this parasite in the absence of domestic dogs, horses and donkeys strongly indicates its transmission in a wildlife cycle. Further, a variety of sequences were obtained from eggs and cysticerci from lions, cheetahs, caracals, spotted hyenas and oryx, which most closely clustered with species of Taenia. Only 3 of them, 2 of lion and 1 of hyena origin, could be allocated to Hydatigera (=Taenia) taeniaeformis (lion), Taenia regis (lions and oryx) and Taenia cf. crocutae (spotted hyena and oryx). PMID:25830103

  13. Filter system for purifying gas or air streams

    International Nuclear Information System (INIS)

    Ohlmeyer, M.; Wilhelm, J.

    1981-01-01

    A filter system is provided for purifying a gas stream by means of flowable or tricklable contact filter material, wherein the stream flows through the filter material and the filter material forms a movable bed. The system contains a filter chamber through which the filter material can flow and which is provided with an inlet opening and an outlet opening for the filter material between which the filter material is conveyed by gravity. The filter system includes deflection means for deflecting the stream , after a first passage of the stream through the filter bed to charge the filter bed for a first time, to a position above where the stream first passed through the filter bed and for conducting the stream at least once again transversely through the filter bed above the first charge so that the filter bed is charged a second time. The filter chamber contains a first opening where the stream enters the filter bed for the first time and is aligned with the deflection means, and a second opening aligned with the deflection means and above the first opening. The second opening is located where the stream leaves the filter bed for the second time, with a partial quantity of the gas stream being able to pass directly through the filter bed from the first opening to the second opening without going through the deflection means. The distance between the upper edge of the first opening and the lower edge of the second opening is at least twice the thickness of the filter chamber

  14. Pyroelectric energy harvesting using Olsen cycles in purified and porous poly(vinylidene fluoride-trifluoroethylene) [P(VDF-TrFE)] thin films

    International Nuclear Information System (INIS)

    Navid, Ashcon; Pilon, Laurent

    2011-01-01

    This paper is concerned with the direct conversion of heat into electricity using pyroelectric materials. The Olsen (or Ericsson) cycle was experimentally performed on three different types of 60/40 poly(vinylidene fluoride-trifluoroethylene) [P(VDF-TrFE)] copolymer samples, namely commercial, purified, and porous films. This cycle consists of two isoelectric field and two isothermal processes. The commercial and purified films were about 50 µm thick and produced a maximum energy density of 521 J l −1 and 426 J l −1 per cycle, respectively. This was achieved by successively dipping the films in cold and hot silicone oil baths at 25 and 110 °C under low and high applied electric fields of about 200 and 500 kV cm −1 , respectively. The 11 µm thick porous films achieved a maximum energy density of 188 J l −1 per cycle between 25 and 100 °C and electric field between 200 and 400 kV cm −1 . The performance of the purified and porous films suffered from their lower electrical resistivity and electric breakdown compared with commercial thin films. However, the energy densities of all 60/40 P(VDF-TrFE) films considered matched or exceeded those reported recently for 0.9Pb(Mg 1/3 Nb 2/3 )O 3 –0.10PbTiO 3 (0.9PMN–0.1PT) (186 J l −1 ) and Pb(Zn 1/3 Nb 2/3 ) 0.955 Ti 0.045 O 3 (243 J l −1 ) bulk ceramics. Furthermore, the results are discussed in light of recently proposed figures of merit for energy harvesting applications

  15. Removal of Inorganic, Microbial, and Particulate Contaminants from a Fresh Surface Water: Village Marine Tec. Expeditionary Unit Water Purifier, Generation 1

    Science.gov (United States)

    The Village Marine Tec. Generation 1 Expeditionary Unit Water Purifier (EUWP) is a mobile skid-mounted system employing ultrafiltration (UF) and reverse osmosis (RO) to produce drinking water from a variety of different water quality sources. The UF components were evaluated to t...

  16. Purified blueberry anthocyanins and blueberry juice alter development of obesity in mice fed an obesogenic high-fat diet.

    Science.gov (United States)

    Prior, Ronald L; E Wilkes, Samuel; R Rogers, Theodore; Khanal, Ramesh C; Wu, Xianli; Howard, Luke R

    2010-04-14

    Male C57BL/6J mice (25 days of age) were fed either a low-fat diet (10% kcal from fat) (LF) or a high-fat diet (45% kcal from fat) (HF45) for a period of 72 days. Blueberry juice or purified blueberry anthocyanins (0.2 or 1.0 mg/mL) in the drinking water were included in LF or HF45 treatments. Sucrose was added to the drinking water of one treatment to test if the sugars in blueberry juice would affect development of obesity. Total body weights (g) and body fat (%) were higher and body lean tissue (%) was lower in the HF45 fed mice compared to the LF fed mice after 72 days, but in mice fed HF45 diet plus blueberry juice or blueberry anthocyanins (0.2 mg/mL), body fat (%) was not different from those mice fed the LF diet. Anthocyanins (ACNs) decreased retroperitoneal and epididymal adipose tissue weights. Fasting serum glucose concentrations were higher in mice fed the HF45 diet. However, it was reduced to LF levels in mice fed the HF45 diet plus 0.2 mg of ACNs/mL in the drinking water, but not with blueberry juice. beta cell function (HOMA-BCF) score was lowered with HF45 feeding but returned to normal levels in mice fed the HF45 diet plus purified ACNs (0.2 mg/mL). Serum leptin was elevated in mice fed HF45 diet, and feeding either blueberry juice or purified ACNs (0.2 mg/mL) decreased serum leptin levels relative to HF45 control. Sucrose in drinking water, when consumption was restricted to the volume of juice consumed, produced lower serum leptin and insulin levels, leptin/fat, and retroperitoneal and total fat (% BW). Blueberry juice was not as effective as the low dose of anthocyanins in the drinking water in preventing obesity. Additional studies are needed to determine factors responsible for the differing responses of blueberry juice and whole blueberry in preventing the development of obesity.

  17. The highly antigenic 53/25 kDa Taenia solium protein fraction with cathepsin-L like activity is present in the oncosphere/cysticercus and induces non-protective IgG antibodies in pigs

    Science.gov (United States)

    Zimic, Mirko; Pajuelo, Mónica; Gilman, Robert H.; Gutiérrez, Andrés H.; Rueda, Luis D.; Flores, Myra; Chile, Nancy; Verástegui, Manuela; Gonzalez, Armando; García, Héctor H.; Sheen, Patricia

    2011-01-01

    Cathepsin L-like proteases are secreted by several parasites including Taenia solium. The mechanism used by T. solium oncospheres to degrade and penetrate the intestine and infect the host is incompletely understood. It is assumed that intestinal degradation is driven by the proteolytic activity of enzymes secreted by the oncosphere. Blocking the proteolytic activity by an antibody response would prevent the oncosphere penetration and further infection. Serine and cysteine proteases including chymotrypsin, trypsin, elastase, and cathepsin L, are secreted by T. solium and Taenia saginata oncospheres when cultured in vitro, being potential vaccine candidates. However, the purification of a sufficient quantity of proteases secreted by oncospheres to conduct a vaccine trial is costly and lengthy. A 53/25 kDa cathepsin L-like fraction partially purified from T. solium cyst fluid was described previously as an important antigen for immunodiagnostics. In this study we found that this antigen is present in the T. solium oncosphere and is also secreted by the cysticercus. This protein fraction was tested for its ability to protect pigs against an oral challenge with T. solium oncospheres in a vaccine trial. IgG antibodies against the 53/25 kDa cathepsin L-like protein fraction were elicited in the vaccinated animals but did not confer protection. PMID:22119017

  18. Current status of human taeniasis in Lao People's Democratic Republic.

    Science.gov (United States)

    Jeon, Hyeong-Kyu; Yong, Tai-Soon; Sohn, Woon-Mok; Chai, Jong-Yil; Min, Duk-Young; Yun, Cheong-Ha; Rim, Han-Jong; Pongvongsa, Tiengkham; Banouvong, Virasack; Insisiengmay, Bounnaloth; Phommasack, Bounlay; Eom, Keeseon S

    2013-04-01

    Human taeniasis was investigated in Lao People's Democratic Republic (Lao PDR) between 2000 and 2011 as part of the nation's helminthiasis survey. A total of 55,038 inhabitants, including 29,846 school children, were examined using the Kato-Katz and scotch-tape anal swab method, and morphological observation of adult worms. Molecular identification of Taenia tapeworms was performed by multiplex PCR or DNA sequence analysis of the mitochondrial cox1 gene. Taenia eggs were present at a rate of 1.5% (845/55,038) in the subject population. Adult tapeworms were identified as T. solium or T. saginata by analyzing the collectable stool specimens (n=126). Three specimens identified as T. solium were found in Luang Prabang, while the remaining 123 specimens, which were T. saginata, were found in Bokeo, Bolikhamxay, Champasak, Houaphan, Khammouane, Luang Namta, Luang Prabang, Oudomxay, Phongsaly, Saysomboune, Saravane, Savannakhet, Xayaboury, Xekong, Xieng Khouang Province, and Vientiane Municipality.

  19. Prophylactic treatment of flea-infested cats with an imidacloprid/flumethrin collar to forestall infection with Dipylidium caninum.

    Science.gov (United States)

    Fourie, Josephus J; Crafford, Dionne; Horak, Ivan G; Stanneck, Dorothee

    2012-07-27

    The objective of the study was to determine the sustained effectiveness of 10% imidacloprid (w/w) and 4.5% flumethrin (w/w) incorporated in a slow-release matrix collar in preventing Dipylidium caninum infection in cats following repeated laboratory-infestations with fleas infected with metacestodes. Efficacy against infection with D. caninum was evaluated by infesting 16 cats with the flea Ctenocephalides felis felis infected with metacestodes of the tapeworm. Medicated collars were fitted to 8 of the cats and infestation of each cat with 200 fleas from a suitably infected batch commenced 7 days later and continued at weekly intervals until Day 28. Efficacy against fleas was evaluated 24 h after each infestation. Infection of the cats with D. caninum was verified by daily examination of the cats' faeces and immediate surroundings for proglottids from Day 21 to Day 60. Calculation of the prophylactic effectiveness of the collars in preventing infection of the cats with D. caninum was based on the difference in the geometric mean number of scoleces recovered from the gastrointestinal tracts of collared compared to untreated cats at necropsy on Day 61. Efficacy of the collars against infestation of the cats with fleas was 99.9% on Day 7 and 100% at each subsequent weekly assessment. Infection of the fleas with metacestodes was ≥40% in 7 to 13 day old fleas, but progressively decreased thereafter. At necropsy all the control cats were infected with D. caninum and harboured between 19 and 346 scoleces with a geometric mean of 58.3. A single treated cat was infected and harboured 2 scoleces. Effective prevention of infection with D. caninum, based on a comparison of the geometric mean numbers of scoleces recovered from control and treated cats, was 99.7%. The insecticidal components of the medicated collars are capable of rapidly eliminating newly-acquired infestations of fleas that are infected with the metacestodes of D. caninum, thus preventing infection with the

  20. Prophylactic treatment of flea-infested dogs with an imidacloprid / flumethrin collar (Seresto®, Bayer) to preempt infection with Dipylidium caninum.

    Science.gov (United States)

    Fourie, Josephus J; Crafford, Dionne; Horak, Ivan G; Stanneck, Dorothee

    2013-08-01

    The objective of the study was to determine the sustained effectiveness of 10 % imidacloprid and 4.5 % flumethrin, incorporated in a slow-release matrix collar, in preventing Dipylidium caninum infection in dogs after repeated laboratory infestations with fleas infected with metacestodes of this tapeworm. Efficacy against infection with D. caninum was evaluated by infesting 16 dogs with cat fleas (Ctenocephalides felis) on study days 7, 14, 21, 28, 35 and 42, from batches suitably infected with D. caninum metacestodes. Prior to each post-treatment infestation the D. caninum infection rate for the fleas was determined by microscopically examining 100 fleas for D. caninum metacestodes. The D. caninum prevalence in the fleas used for infestations ranged from 23 % to 52 %. Medicated collars were fitted to 8 of the dogs on study day 0. The weight of the IVP collars varied between 35.48 g and 38.48 g (average 37.16 g), whilst animal weight varied between 12.20 kg and 17.98 kg (treated group, n = 8, average 14.79 kg). Seven days later infestation of each of the 16 dogs with 250 fleas commenced. Infestations continued at weekly intervals until Day 42 with efficacy against fleas evaluated 24 hours after each infestation. From Days 21 to 74, infection of the dogs with D. caninum was verified (daily examination of faeces and cages for the presence of expelled proglottids). Calculation of prophylactic effectiveness of the collars in preventing infection with D. caninum was based on the difference in geometric mean numbers of scoleces between groups at necropsy on Day 75. Effective prevention of infection with D. caninum was found to be 96.6 %. Efficacy of the collars against fleas was ≥ 99.9 % for the duration of the assessment period. Newly acquired infestations of fleas are rapidly eliminated by the insecticidal components of the medicated collars over a period of several months. In the event of fleas being infected with metacestodes, with D. caninum can be prevented in

  1. Prophylactic treatment of flea-infested cats with an imidacloprid/flumethrin collar to forestall infection with Dipylidium caninum

    Directory of Open Access Journals (Sweden)

    Fourie Josephus J

    2012-07-01

    Full Text Available Abstract Background The objective of the study was to determine the sustained effectiveness of 10% imidacloprid (w/w and 4.5% flumethrin (w/w incorporated in a slow-release matrix collar in preventing Dipylidium caninum infection in cats following repeated laboratory-infestations with fleas infected with metacestodes. Methods Efficacy against infection with D. caninum was evaluated by infesting 16 cats with the flea Ctenocephalides felis felis infected with metacestodes of the tapeworm. Medicated collars were fitted to 8 of the cats and infestation of each cat with 200 fleas from a suitably infected batch commenced 7 days later and continued at weekly intervals until Day 28. Efficacy against fleas was evaluated 24 h after each infestation. Infection of the cats with D. caninum was verified by daily examination of the cats’ faeces and immediate surroundings for proglottids from Day 21 to Day 60. Calculation of the prophylactic effectiveness of the collars in preventing infection of the cats with D. caninum was based on the difference in the geometric mean number of scoleces recovered from the gastrointestinal tracts of collared compared to untreated cats at necropsy on Day 61. Results Efficacy of the collars against infestation of the cats with fleas was 99.9% on Day 7 and 100% at each subsequent weekly assessment. Infection of the fleas with metacestodes was ≥40% in 7 to 13 day old fleas, but progressively decreased thereafter. At necropsy all the control cats were infected with D. caninum and harboured between 19 and 346 scoleces with a geometric mean of 58.3. A single treated cat was infected and harboured 2 scoleces. Effective prevention of infection with D. caninum, based on a comparison of the geometric mean numbers of scoleces recovered from control and treated cats, was 99.7%. Conclusion The insecticidal components of the medicated collars are capable of rapidly eliminating newly-acquired infestations of fleas that are infected

  2. Multielemental segregation analysis of the thallium bromide impurities purified by repeated Bridgman technique

    International Nuclear Information System (INIS)

    Santos, Robinson A. dos; Hamada, Margarida M.; Costa, Fabio E. da; Gennari, Roseli F.; Martins, Joao F.T.; Marcondes, Renata M.; Mesquita, Carlos H. de

    2011-01-01

    TlBr crystals were purified and grown by the repeated Bridgman method from two commercial TlBr salts and characterized to be used as radiation detectors. To evaluate the purification efficiency, measurements of the impurity concentration were made after each growth, analyzing the trace impurities by inductively coupled plasma mass spectroscopy (ICP-MS). A significant decrease of the impurity concentration resulting from the purification number was observed. To evaluate the crystal as a radiation semiconductor detector, measurements of its resistivity and gamma-ray spectroscopy were carried out. The radiation response depended on the crystal purity. The repeated Bridgman technique improved the TlBr crystal quality used as a radiation detector. A compartmental model was proposed to fit the impurity concentration as a function of the repetition number of the Bridgman growth. (author)

  3. Performance Variation of Spent Resin in Mixed Bed From Water Purifying System of Xi'an Pulse Reactor

    International Nuclear Information System (INIS)

    Li Hua; Ma Yan; Xiao Yan; Liu Yueheng; Yang Yongqing

    2010-01-01

    Detailed physical and chemical characteristic analysis was performed on the spent cation and anion resins in the mixed bed from Xi'an Pulse Reactor water purifying system.The exchange performance variations of used resins and the contributions from different factors to the variation were discussed.Based on the obtained information of the impurities in the used resin, the contamination state of the water in the Xi'an Pulse Reactor water pool, the corrosion state of the structural material in the reactor was presented. The spent anion resin almost completely losses its exchange performance,while the remaining exchange capacity in the spent cation resin is still high.The radiation field from the reactor operation contributes little to the degradation of the performance of the resins. The exchange capacity loss of the spent anion resin is due to the exchange of its active groups into abundant carbonate and a certain amount of organics. The impurity amount in the anion and cation exchange resins is low,which suggests(that) the water in the Xi'an Pulse Reactor water pool is little contaminated. A certain extent of corrosion is occurred on the structural material in the swimming pool of the reactor. The results provide important referential data for the operational safety of the water purifying system of similar research reactor. (authors)

  4. Metabolism of benzene and phenol by a reconstituted purified phenobarbital induced rat liver mixed function oxidase system

    International Nuclear Information System (INIS)

    Griffiths, J.C.

    1986-01-01

    Cytochrome P-450 and the electron-donor, NADPH-cytochrome c reductase were isolated from phenobarbital induced rat liver microsomes. Both benzene and its primary metabolite phenol, were substrates for the reconstituted purified phenobarbital induced rat liver mixed function oxidase system. Benzene was metabolized to phenol and the polyhydroxylated metabolites; catechol, hydroquinone and 1,2,4 benzenetriol. Benzene elicited a Type I spectral change upon its interaction with the cytochrome P-450 while phenol's interaction with the cytochrome P-450 produced a reverse Type I spectra. The formation of phenol showed a pH optimum of 7.0 compared with 6.6-6.8 for the production of the polyhyrdoxylated metabolites. Cytochrome P-450 inhibitors, such as metyrapone and SKF 525A, diminished the production of phenol from benzene but not the production of the polyhydroxylated metabolites from phenol. The radical trapping agents, DMSO, KTBA and mannitol, decreased the recovery of polyhydroxylated metabolites, from 14 C-labeled benzene and/or phenol. As KTBA and DMSO interacted with OH. There was a concomitant release of ethylene and methane, which was measured. Desferrioxamine, an iron-chelator and catalase also depressed the recovery of polyhydroxylated metabolites. In summary, benzene and phenol were both substrates for this reconstituted purified enzyme system, but they differed in binding to cytochrome P-450, pH optima and mode of hydroxylation

  5. Cysticercosis in cattle and its public health implications in Mekelle ...

    African Journals Online (AJOL)

    animals highlights the public health significance of this infection. Key words: Cysticercosis, Cysticercus bovis, occurrence, taeniasis, Taenia saginata,. Introduction. Bovine cysticercosis is the infection of the cattle caused by Cysticercus bovis and consumption of the raw or undercooked meat results in human taeniasis. Also ...

  6. Browse Title Index

    African Journals Online (AJOL)

    ... Spectrum of neurosurgical complications following medical tourism: challenges of ... Vol 13, No 3 (2013), Steroid induced diabetes mellitus in patients receiving .... Vol 15, No 1 (2015), Taenia solium metacestode preparation in rural areas of ...

  7. Oligomer formation and G-quadruplex binding by purified murine Rif1 protein, a key organizer of higher-order chromatin architecture.

    Science.gov (United States)

    Moriyama, Kenji; Yoshizawa-Sugata, Naoko; Masai, Hisao

    2018-03-09

    Rap1-interacting protein 1 (Rif1) regulates telomere length in budding yeast. We previously reported that, in metazoans and fission yeast, Rif1 also plays pivotal roles in controlling genome-wide DNA replication timing. We proposed that Rif1 may assemble chromatin compartments that contain specific replication-timing domains by promoting chromatin loop formation. Rif1 also is involved in DNA lesion repair, restart after replication fork collapse, anti-apoptosis activities, replicative senescence, and transcriptional regulation. Although multiple physiological functions of Rif1 have been characterized, biochemical and structural information on mammalian Rif1 is limited, mainly because of difficulties in purifying the full-length protein. Here, we expressed and purified the 2418-amino-acid-long, full-length murine Rif1 as well as its partially truncated variants in human 293T cells. Hydrodynamic analyses indicated that Rif1 forms elongated or extended homo-oligomers in solution, consistent with the presence of a HEAT-type helical repeat segment known to adopt an elongated shape. We also observed that the purified murine Rif1 bound G-quadruplex (G4) DNA with high specificity and affinity, as was previously shown for Rif1 from fission yeast. Both the N-terminal (HEAT-repeat) and C-terminal segments were involved in oligomer formation and specifically bound G4 DNA, and the central intrinsically disordered polypeptide segment increased the affinity for G4. Of note, pulldown assays revealed that Rif1 simultaneously binds multiple G4 molecules. Our findings support a model in which Rif1 modulates chromatin loop structures through binding to multiple G4 assemblies and by holding chromatin fibers together. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Influence of thermodynamic mechanism of inter- facial adsorption on purifying air-conditioning engineering under intensification of electric field

    Directory of Open Access Journals (Sweden)

    Chen Yun-Yu

    2016-12-01

    Full Text Available As a kind of mass transfer process as well as the basis of separating and purifying mixtures, interfacial adsorption has been widely applied to fields like chemical industry, medical industry and purification engineering in recent years. Influencing factors of interfacial adsorption, in addition to the traditional temperature, intensity of pressure, amount of substance and concentration, also include external fields, such as magnetic field, electric field and electromagnetic field, etc. Starting from the point of thermodynamics and taking the Gibbs adsorption as the model, the combination of energy axiom and the first law of thermodynamics was applied to boundary phase, and thus the theoretical expression for the volume of interface absorption under electric field as well as the mathematical relationship between surface tension and electric field intensity was obtained. In addition, according to the obtained theoretical expression, the volume of interface absorption of ethanol solution under different electric field intensities and concentrations was calculated. Moreover, the mechanism of interfacial adsorption was described from the perspective of thermodynamics and the influence of electric field on interfacial adsorption was explained reasonably, aiming to further discuss the influence of thermodynamic mechanism of interfacial adsorption on purifying air-conditioning engineering under intensification of electric field.

  9. Purifier-integrated methanol reformer for fuel cell vehicles

    Science.gov (United States)

    Han, Jaesung; Kim, Il-soo; Choi, Keun-Sup

    We developed a compact, 3-kW, purifier-integrated modular reformer which becomes the building block of full-scale 30-kW or 50-kW methanol fuel processors for fuel cell vehicles. Our proprietary technologies regarding hydrogen purification by composite metal membrane and catalytic combustion by washcoated wire-mesh catalyst were combined with the conventional methanol steam-reforming technology, resulting in higher conversion, excellent quality of product hydrogen, and better thermal efficiency than any other systems using preferential oxidation. In this system, steam reforming, hydrogen purification, and catalytic combustion all take place in a single reactor so that the whole system is compact and easy to operate. Hydrogen from the module is ultrahigh pure (99.9999% or better), hence there is no power degradation of PEMFC stack due to contamination by CO. Also, since only pure hydrogen is supplied to the anode of the PEMFC stack, 100% hydrogen utilization is possible in the stack. The module produces 2.3 Nm 3/h of hydrogen, which is equivalent to 3 kW when PEMFC has 43% efficiency. Thermal efficiency (HHV of product H 2/HHV of MeOH in) of the module is 89% and the power density of the module is 0.77 kW/l. This work was conducted in cooperation with Hyundai Motor Company in the form of a Korean national project. Currently the module is under test with an actual fuel cell stack in order to verify its performance. Sooner or later a full-scale 30-kW system will be constructed by connecting these modules in series and parallel and will serve as the fuel processor for the Korean first fuel cell hybrid vehicle.

  10. Bitistatin-functionalized fluorescent nanodiamond particles specifically bind to purified human platelet integrin receptor αIIbβ3 and activated platelets.

    Science.gov (United States)

    Marcinkiewicz, Cezary; Gerstenhaber, Jonathan A; Sternberg, Mark; Lelkes, Peter I; Feuerstein, Giora

    2017-01-01

    Thromboembolic events (TEE) underwrite key causes of death in developed countries. While advanced imaging technologies such as computed tomography scans serve to diagnose blood clots during acute cardiovascular events, no such technology is available in routine primary care for TEE risk assessment. Here, we describe an imaging platform technology based on bioengineered fluorescent nanodiamond particles (F-NDPs) functionalized with bitistatin (Bit), a disintegrin that specifically binds to the α IIb β 3 integrin, platelet fibrinogen receptor (PFR) on activated platelets. Covalent linkage of purified Bit to F-NDP was concentration-dependent and saturable, as validated by enzyme-linked immunosorbent assay using specific anti-Bit antibodies. F-NDP-Bit interacted with purified PFR, either in immobilized or soluble form. Lotrafiban, a nonpeptide, α IIb β 3 receptor antagonist, specifically blocked F-NDP-Bit-PFR complex formation. Moreover, F-NDP-Bit specifically binds to activated platelets incorporated into a clot generated by thrombin-activated rat platelet-rich plasma (PRP). Our results suggest that engineered F-NDP-Bit particles could serve as noninvasive, "real-time" optical diagnostics for clots present in blood vessels.

  11. Utilization of highly purified single wall carbon nanotubes dispersed in polymer thin films for an improved performance of an electrochemical glucose sensor

    Energy Technology Data Exchange (ETDEWEB)

    Goornavar, Virupaxi [Molecular Toxicology Laboratory, Center for Biotechnology and Biomedical Sciences, Norfolk State University, 700 Park Avenue, Norfolk, VA 23504 (United States); Center for Materials Research, Norfolk State University, 555 Park Avenue, Norfolk, VA 23504 (United States); Jeffers, Robert [Molecular Toxicology Laboratory, Center for Biotechnology and Biomedical Sciences, Norfolk State University, 700 Park Avenue, Norfolk, VA 23504 (United States); Luna Innovations, Inc., 706 Forest St., Suite A, Charlottesville, VA 22902 (United States); Biradar, Santoshkumar [RICE University, 6100 Main St, Houston, TX 77251 (United States); Ramesh, Govindarajan T., E-mail: gtramesh@nsu.edu [Molecular Toxicology Laboratory, Center for Biotechnology and Biomedical Sciences, Norfolk State University, 700 Park Avenue, Norfolk, VA 23504 (United States); Center for Materials Research, Norfolk State University, 555 Park Avenue, Norfolk, VA 23504 (United States)

    2014-07-01

    In this work we report the improved performance an electrochemical glucose sensor based on a glassy carbon electrode (GCE) that has been modified with highly purified single wall carbon nanotubes (SWCNTs) dispersed in polyethyleneimine (PEI), polyethylene glycol (PEG) and polypyrrole (PPy). The single wall carbon nanotubes were purified by both thermal and chemical oxidation to achieve maximum purity of ∼ 98% with no damage to the tubes. The SWCNTs were then dispersed by sonication in three different organic polymers (1.0 mg/ml SWCNT in 1.0 mg/ml of organic polymer). The stable suspension was coated onto the GCE and electrochemical characterization was performed by Cyclic Voltammetry (CV) and Amperometry. The electroactive enzyme glucose oxidase (GOx) was immobilized on the surface of the GCE/(organic polymer–SWCNT) electrode. The amperometric detection of glucose was carried out at 0.7 V versus Ag/AgCl. The GCE/(SWCNT–PEI, PEG, PPY) gave a detection limit of 0.2633 μM, 0.434 μM, and 0.9617 μM, and sensitivities of 0.2411 ± 0.0033 μA mM{sup −1}, r{sup 2} = 0.9984, 0.08164 ± 0.001129 μA mM{sup −1}, r{sup 2} = 0.9975, 0.04189 ± 0.00087 μA mM{sup −1}, and r{sup 2} = 0.9944 respectively and a response time of less than 5 s. The use of purified SWCNTs has several advantages, including fast electron transfer rate and stability in the immobilized enzyme. The significant enhancement of the SWCNT modified electrode as a glucose sensor can be attributed to the superior conductivity and large surface area of the well dispersed purified SWCNTs. - Highlights: • Purification method employed here use cheap and green oxidants. • The method does not disrupt the electronic structure of nanotubes. • This method removes nearly < 2% metallic impurities. • Increases the sensitivity and performance of glassy carbon electrode • This system can detect as low as 0.066 μM of H{sub 2}O{sub 2} and 0.2633 μM of glucose.

  12. Purified umbilical cord derived mesenchymal stem cell treatment in a case of systemic lupus erythematosus.

    Science.gov (United States)

    Phillips, Christopher D; Wongsaisri, Pornpatcharin; Htut, Thein; Grossman, Terry

    2017-12-01

    Systemic lupus erythematosus (SLE) is a multiple organ system autoimmune disorder for which there is no known cure. We report a case of a young adult lady with SLE and Sjogren's with diagnostic and clinical resolution following purified umbilical cord derived mesenchymal stem cell (MSC) and globulin component protein macrophage activating factor (GcMAF) therapy in a combined multidisciplinary integrative medicine protocol. Our patient had complete reversal of all clinical and laboratory markers. We recommend a prospective randomized double blind study to assess the sustained efficacy of MSC and GcMAF in the treatment of autoimmune connective tissue diseases such as systemic lupus erythematosus.

  13. Method of purifying zirconium tetrachloride and hafnium tetrachloride in a vapor stream

    International Nuclear Information System (INIS)

    Snyder, T.S.; Stolz, R.A.

    1992-01-01

    This patent describes a method of purifying zirconium tetrachloride and hafnium tetrachloride in a vapor stream from a sand chlorinator in which the silicon and metals present in sand fed to the chlorinator are converted to chlorides at temperatures over about 800 degrees C. It comprises cooling a vapor stream from a sand chlorinator, the vapor stream containing principally silicon tetrachloride, zirconium tetrachloride, and hafnium tetrachloride contaminated with ferric chloride, to a temperature of from about 335 degrees C to about 600 degrees C; flowing the vapor stream through a gaseous diffusion separative barrier to produce a silicon tetrachloride-containing vapor stream concentrated in zirconium tetrachloride and hafnium tetrachloride and a silicon tetrachloride-containing vapor stream depleted in zirconium tetrachloride and hafnium tetrachloride; adsorbing the ferric chloride in the separative barrier; and recovering the silicon tetrachloride stream concentrated in zirconium tetrachloride and hafnium tetrachloride separately from the silicon tetrachloride stream depleted in zirconium tetrachloride and hafnium tetrachloride

  14. Combined measurement of growth and differentiation in suspension cultures of purified human CD34-positive cells enables a detailed analysis of myelopoiesis

    NARCIS (Netherlands)

    Kerst, J. M.; Slaper-Cortenbach, I. C.; von dem Borne, A. E.; van der Schoot, C. E.; van Oers, R. H.

    1992-01-01

    In this study we have made a detailed analysis of growth factor (granulocyte-macrophage colony-stimulating factor [GM-CSF], granulocyte colony-stimulating factor [G-CSF], and macrophage colony-stimulating factor [M-CSF])-induced proliferation and differentiation of highly purified CD34+ committed

  15. Inhibition of Epithelial TNF-α Receptors by Purified Fruit Bromelain Ameliorates Intestinal Inflammation and Barrier Dysfunction in Colitis

    OpenAIRE

    Zhou, Zijuan; Wang, Liang; Feng, Panpan; Yin, Lianhong; Wang, Chen; Zhi, Shengxu; Dong, Jianyi; Wang, Jingyu; Lin, Yuan; Chen, Dapeng; Xiong, Yongjian; Peng, Jinyong

    2017-01-01

    Activation of the TNF-α receptor (TNFR) leads to an inflammatory response, and anti-TNF therapy has been administered to reduce inflammation symptoms and heal mucosal ulcers in inflammatory bowel disease (IBD). Bromelain, a complex natural mixture of proteolytic enzymes, has been shown to exert anti-inflammatory effects. This study aimed to investigate the effect of purified fruit bromelain (PFB)-induced inhibition of epithelial TNFR in a rat colitis model. Colitis was established by intracol...

  16. Properties of Folate Binding Protein Purified from Cow’s Milk

    Directory of Open Access Journals (Sweden)

    SUBANDRATE

    2012-09-01

    Full Text Available Folic acid played an important role in the metabolism of the body. To measure the serum folic acid levels could use the folate binding protein (FBP from cow’s milk with a technique analogous to ELISA. The aims of this study were to identify characteristics of FBP from cow’s milk and binding capacity of FBP to folic acid and to purify FBP from other whey protein passed through DEAE-cellulose chromatography column. Each of DEAE-cellulose peaks was passed in affinity chromatography column. FBP was released from affinity column with sodium acetate buffer pH 3.5. The purity of obtained FBP was demonstrated by a single spot in SDS-PAGE analysis and the estimated molecular weight of FBP was around 31 kDa. Our study indicated that 1 mol FBP bound 1 mol folic acid. Alkylation with iodoacetic acid decreased the binding capacity of FBP which suggested the presence of a–SH or imidazol group in its active site. The importance of disulfide bridge was proven by decreasing of folate binding capacity of FBP after -mercaptoethanol treatment. In contrary, the folate binding didn need Ca2+ ion, as indicated by EDTA test which gave the same result as control.

  17. PURIFIED WASTE FCC CATALYST AS A CEMENT REPLACEMENT MATERIAL

    Directory of Open Access Journals (Sweden)

    Danute Vaiciukyniene

    2015-06-01

    Full Text Available Zeolites are commonly used in the fluid catalytic cracking process. Zeolite polluted with oil products and became waste after some time used. The quantity of this waste inevitably rises by expanding rapidly oil industry. The composition of these catalysts depends on the manufacturer and on the process that is going to be used. The main factors retarding hydration process of cement systems and modifying them strength are organic compounds impurities in the waste FCC catalyst. The present paper shows the results of using purified waste FCC catalyst (pFCC from Lithuania oil refinery, as Portland cement replacement material. For this purpose, the purification of waste FCC catalyst (FCC samples was treated with hydrogen peroxide. Hydrogen peroxide (H2O2 is one of the most powerful oxidizers known. By acting of waste with H2O2 it can eliminate the aforementioned waste deficiency, and the obtained product becomes one of the most promising ingredients, in new advanced building materials. Hardened cement paste samples with FCC or pFCC were formed. It was observed that the pFCC blended cements developed higher strength, after 28 days, compared to the samples with FCC or reference samples. Typical content of Portland cement substituting does not exceed 30 % of mass of Portland cement in samples. Reducing the consumption of Portland cement with utilizing waste materials is preferred for reasons of environmental protection.

  18. The correlation between yielding behavior and precipitation in ultra purified ferritic stainless steels

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Z.Y., E-mail: zyliu@mail.neu.edu.cn [State Key Lab of Rolling Technologies and Automation, Northeastern University, Heping Qu, Wenhua St, P.O. Box 105, Shenyang, Liaoning Province 110004 (China); Gao, F. [State Key Lab of Rolling Technologies and Automation, Northeastern University, Heping Qu, Wenhua St, P.O. Box 105, Shenyang, Liaoning Province 110004 (China); Jiang, L.Z. [Research Institute for Stainless Steels, R and D Center, Baosteel Co., Shanghai 201900 (China); Wang, G.D. [State Key Lab of Rolling Technologies and Automation, Northeastern University, Heping Qu, Wenhua St, P.O. Box 105, Shenyang, Liaoning Province 110004 (China)

    2010-06-25

    Cold rolled sheets of a ultra purified ferritic stainless steel were annealed either by being slowly cooled from 950 deg. C or being rapidly cooled to room temperature from the intermediate holding at 750 deg. C. The former exhibited substantial Lueders elongation during tensile testing, while the later showed continuous yielding behavior. In the slowly cooled sheet, both Nb(C, N) and (Fe, Cr){sub 2}Nb have been formed, and no (Fe, Cr){sub 2}Nb could be observed in the rapidly cooled sheet. The fast growth of (Fe, Cr){sub 2}Nb is believed to have caused local depletion of Nb atoms around fine NbC particles, resulting in their dissolution and having carbon atoms released for the formation of the Cottrell atmosphere. These results have been confirmed by the internal friction measurements and thermodynamic calculations.

  19. Radioimmunoassay kit formulation and its validation for serum progesterone using progesterone radiotracer purified by gel filtration

    International Nuclear Information System (INIS)

    Karir, T.; Pal, N.; Sivaprasad, N.

    2003-01-01

    Purification of the radioiodinated progesterone tyrosine methyl ester conjugate by gel filtration and the development, optimization and clinical validation of a direct radioimmunoassay (RIA) of progesterone using this radiotracer are described. High purity radiotracer is essential for the error free performance of any RIA. Progesterone 11α hemisuccinate was conjugated to tyrosine methyl ester (TME) by the mixed anhydride method and this conjugate was then radioiodinated by the chloramine-T method. Purification of the radioiodinated product was carried out by gel filtration. About 12 batches of the radiotracer were prepared and purified. The purification by gel filtration gave reproducible elution pattern and purity. The radiotracer thus purified was found to have consistent quality as compared to that of any other purification methods. Non-specific binding of the radiotracer was found to be 95% as checked by paper electrophoresis. The stability (retention of the immunoreactivity) of the radiotracer was two to three months. No appreciable changes in the assay characteristics were observed during this period. The assay involved 3 hours incubation of progesterone antibody with individual standards or sample and radiotracer at room temperature. The optimized assay was then validated for internal and external quality control parameters. A RIA kit was then formulated with this radiotracer for estimation of progesterone in serum. The assay kit consisted of lyophilized individual standards ranging from 0.25 to 50 ng/ml. The clinical performance of the developed kit was compared with that of a commercial ELISA kit and a correlation of 0.94 was observed. (author)

  20. Strategic of Applying Free Chemical Usage In Purified Water System For Pharmaceutical Industry Toward CPOB (Cara Pembuatan Obat yang Baik Indonesia To Reducing Environmental Pollution

    Directory of Open Access Journals (Sweden)

    Kartono R.

    2014-03-01

    Full Text Available The purpose of this paper is to examine the sets of model and literature review to prove that strategy of applying free chemical usage in purified water system for pharmaceutical industry would be help the existing and new pharmaceutical companies to comply with part of Natioanal Agency of Drug and Food Control / Badan Pengawas Obat dan Makanan (NADFC/BPOM regulation in order to achieve “Cara Pembuatan Obat yang Baik” (CPOB of Indonesia pharmaceutical industry. One of the main reasons is when we figured out the number of Indonesian pharmaceutical industries in 2012 are kept reducing compare to the increasing numbers of Indonesian population growth. This strategy concept also might help the industries to reducing environmental pollution, and operational cost in pharmaceutical industries, by reducing of the chemical usage for water treatment process in floculation and cougulation and chlorination for sterillization. This new model is free usage of chemicals for purified water generation system process and sterilization. The concept offering of using membrane technology- Reverse Osmosis (RO membrane base treatment to replace traditional chemical base treatment, following enhance Electrodeionization (EDI as final polisher for controlling conductivity, and finally Ultra Violet (UV disinfectant technology as final guard for bacteria controls instead of chemical base system in purified water generation system.

  1. Strategic of Applying Free Chemical Usage In Purified Water System For Pharmaceutical Industry Toward CPOB (Cara Pembuatan Obat yang Baik) Indonesia To Reducing Environmental Pollution

    Science.gov (United States)

    Kartono, R.; Basuki, Y. T.

    2014-03-01

    The purpose of this paper is to examine the sets of model and literature review to prove that strategy of applying free chemical usage in purified water system for pharmaceutical industry would be help the existing and new pharmaceutical companies to comply with part of Natioanal Agency of Drug and Food Control / Badan Pengawas Obat dan Makanan (NADFC/BPOM) regulation in order to achieve "Cara Pembuatan Obat yang Baik" (CPOB) of Indonesia pharmaceutical industry. One of the main reasons is when we figured out the number of Indonesian pharmaceutical industries in 2012 are kept reducing compare to the increasing numbers of Indonesian population growth. This strategy concept also might help the industries to reducing environmental pollution, and operational cost in pharmaceutical industries, by reducing of the chemical usage for water treatment process in floculation and cougulation and chlorination for sterillization. This new model is free usage of chemicals for purified water generation system process and sterilization. The concept offering of using membrane technology- Reverse Osmosis (RO) membrane base treatment to replace traditional chemical base treatment, following enhance Electrodeionization (EDI) as final polisher for controlling conductivity, and finally Ultra Violet (UV) disinfectant technology as final guard for bacteria controls instead of chemical base system in purified water generation system.

  2. Simultaneously upgrading biogas and purifying biogas slurry using cocultivation of Chlorella vulgaris and three different fungi under various mixed light wavelength and photoperiods.

    Science.gov (United States)

    Cao, Weixing; Wang, Xue; Sun, Shiqing; Hu, Changwei; Zhao, Yongjun

    2017-10-01

    In order to purify biogas slurry and biogas simultaneously, three different fungi, Pleurotus geesteranus (P. geesteranus), Ganoderma lucidum (G. lucidum), and Pleurotus ostreatus (P. ostreatus) were pelletized with Chlorella vulgaris (C. vulgaris). The results showed that the optimal light wavelength ratio for red:blue was 5:5 for these three different fungi-assisted C. vulgaris, resulting in higher specific growth rate as well as nutrient and CO 2 removal efficiency compared with other ratios. G. lucidum/C. vulgaris was screened as the best fungi-mialgae for biogas slurry purification and biogas upgrading with light/dark ratio of 14h:10h, which was also confirmed by the economic efficiency analysis of the energy consumptions. These results will provide a theoretical foundation for large-scale biogas slurry purifying and biogas upgrading using microalgae. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Molecular identification of species of Taenia causing bovine cysticercosis in Ethiopia.

    Science.gov (United States)

    Hailemariam, Z; Nakao, M; Menkir, S; Lavikainen, A; Iwaki, T; Yanagida, T; Okamoto, M; Ito, A

    2014-09-01

    Bovine cysticercosis causing damage to the beef industry is closely linked to human taeniasis due to Taenia saginata. In African countries, Taenia spp. from wildlife are also involved as possible sources of infections in livestock. To identify the aetiological agents of bovine cysticercosis in Ethiopia, cysticerci were collected from 41 cattle slaughtered in the eastern and central areas during 2010-2012. A single cysticercus per animal was subjected to the polymerase chain reaction (PCR)-based DNA sequencing of mitochondrial cytochrome c oxidase subunit 1 gene, and the resultant sequence was compared with those of members of the genus Taenia. Although 38 out of 41 cysticerci (92.7%) were identified as T. saginata, three samples (7.3%) showed the hitherto unknown sequences of Taenia sp., which is distantly related to Taenia solium, Taenia arctos and Taenia ovis. Old literatures suggest it to be Taenia hyaenae, but morphological identification of species could not be completed by observing only the larval samples.

  4. [Effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes in the murine model of vaginal candidiasis].

    Science.gov (United States)

    Wang, F; Huo, Y; Yin, L R; Sun, B; Zhang, P P

    2016-07-25

    To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. (1)Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis(VVC)murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group(n=15)and control group(n=15). Suspension(30 μl, 100 μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline(PBS)was given to the control group.(2)Tweenty-four hours after treatment, the fungal burden and colony-forming unit(CFU)of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. (1)VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group.(2)Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×10(4) CFU/ml]than that in the control group [(8.5±2.1)×10(4) CFU/ml, P=0.017]. IFN-γ level of the test group was increased [(257±11)vs(197±4)pg/ml, P=0.000], while the level of IL-10 was reduced [(287 ± 15)vs(379 ± 17)pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group(0.892±0.008 vs 0.496±0.013, P=0.000). Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γ and IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37

  5. Characterization of 14-3-3 isoforms expressed in the Echinococcus granulosus pathogenic larval stage.

    Science.gov (United States)

    Teichmann, Aline; Vargas, Daiani M; Monteiro, Karina M; Meneghetti, Bruna V; Dutra, Cristine S; Paredes, Rodolfo; Galanti, Norbel; Zaha, Arnaldo; Ferreira, Henrique B

    2015-04-03

    The 14-3-3 protein family of eukaryotic regulators was studied in Echinococcus granulosus, the causative agent of cystic hydatid disease. These proteins mediate important cellular processes in eukaryotes and are expected to play important roles in parasite biology. Six isoforms of E. granulosus 14-3-3 genes and proteins (Eg14-3-3.1-6) were analyzed, and their phylogenetic relationships were established with bona fide 14-3-3 orthologous proteins from eukaryotic species. Eg14-3-3 isoforms with previous evidence of expression (Eg14-3-3.1-4) in E. granulosus pathogenic larval stage (metacestode) were cloned, and recombinant proteins were used for functional studies. These protein isoforms were detected in different components of E. granulosus metacestode, including interface components with the host. The roles that are played by Eg14-3-3 proteins in parasite biology were inferred from the repertoires of interacting proteins with each isoform, as assessed by gel overlay, cross-linking, and affinity chromatography assays. A total of 95 Eg14-3-3 protein ligands were identified by mass spectrometry. Eg14-3-3 isoforms have shared partners (44 proteins), indicating some overlapping functions; however, they also bind exclusive partners (51 proteins), suggesting Eg14-3-3 functional specialization. These ligand repertoires indicate the involvement of Eg14-3-3 proteins in multiple biochemical pathways in the E. granulosus metacestode and note some degree of isoform specialization.

  6. Characterization of purified α-amylase produced by Aspergillus terreus NCFT 4269.10 using pearl millet as substrate

    Directory of Open Access Journals (Sweden)

    Bijay Kumar Sethi

    2016-12-01

    Full Text Available α-amylase was produced by Aspergillus terreus NCFT 4269.10 using both liquid static surface (LSSF and solid-state fermentation using pearl millet residues as substrate. The maximum production of α-amylase was noticed at 30°C incubated for 96h. The crude α-amylase was purified to electrophoretic homogeneity and characterized. Characterization of amylase confirmed that the purified α-amylase was found to be most stable at pH 5.0, 60°C temperature, and a substrate concentration of 1.25%. The enzyme was active for 40 min at 70°C with an optimum enzyme–substrate reaction time of 60 min. Amylase was compatible with all detergents tested having highest activity with Surf excel followed by Henko and Ariel. SDS and Tween 20 reduced the activity. Among the metal ions tested, the maximum α-amylase activity was attained in the presence of Ca2+, followed by Mg2+ and Mn2+. The activity of α-amylase was not considerably affected in the presence of ethylenediaminetetraacetic acid and Triton X-100. Amylase activity was accelerated in the presence of sodium lauryl sulfate and phenylmethylsulfonyl fluoride did not significantly (or slightly affect the activity and stability. Tween 20, urea (5%, and the reducing agent, β-mercaptoethanol significantly inhibited the activity of α-amylase. Owing to its noteworthy stability in the presence of detergents, additives, inhibitors, and metal ions, this α-amylase could be an impending enzyme for significant industrial exploitations.

  7. Regenerated silica gel as stationary phase on vacuum column chromatography to purify temulawak’s extracts

    International Nuclear Information System (INIS)

    Cahyono, Bambang; Maduwu, Ratna Dewi; Widayat,; Suzery, Meiny

    2015-01-01

    Commercial silica gel only used once by many researchers and affected high cost for purification process, also less support the green chemistry program. This research focused in regeneration silica gel that used purification of temulawak’s extracts (Curcuma xanthorrhiza Roxb) by vacuum column chromatography. Sample extracts (contains 10.1195±0.5971% of curcuminoids) was purified by vacuum column chromatography (pressure: 45 kPa, column: 100mm on length and 16mm on diameter). Ethanol 96% and acetone were compared as eluent. The amount of solvent and yield of curcuminoids used as indicator purification. The silica gel was regenerated with heating in 600°C for 8 hours The silica gels were analyzed by IR spectroscopy and X-ray diffraction. Furthermore, regenerated silica gel was used as the stationary phase in vacuum column chromatography under the same conditions with the previous purification. All the purification experiments were performed in three repetitions. Based on regression equation, y=0.132x+0.0011 (r 2 =0.9997) the yield of curcuminoids on purified products using ethanol as the eluent was improved 4.26% (to 14.3724±0.5749%) and by acetone was improved 3,03% (to 13.1450 ±0.6318%). The IR spectrum of both silica gel showed the same vibration profile and also there were three crystallinity peaks missing on its X-ray diffraction. Regenerated silica gel has the same performance with new silica gel in purification of temulawak’s extract: by ethanol has increased 4.08% (14.1947±0.7415%) and 2.93% (13.0447±0.4822) by acetone. In addition, all purification products showed similar TLC profiles. Purification using regenerated silica gel as the adsorbent on vacuum column chromatography has exactly same potential with the new silica gel

  8. Regenerated silica gel as stationary phase on vacuum column chromatography to purify temulawak’s extracts

    Energy Technology Data Exchange (ETDEWEB)

    Cahyono, Bambang; Maduwu, Ratna Dewi; Widayat,; Suzery, Meiny [Organic Chemistry Laboratory, Departement of Chemistry, Diponegoro University Jln Prof. Soedharto SH, Tembalang, Semarang 50275, Indonesia Tel / Fax: (024) 7460058 (Indonesia)

    2015-12-29

    Commercial silica gel only used once by many researchers and affected high cost for purification process, also less support the green chemistry program. This research focused in regeneration silica gel that used purification of temulawak’s extracts (Curcuma xanthorrhiza Roxb) by vacuum column chromatography. Sample extracts (contains 10.1195±0.5971% of curcuminoids) was purified by vacuum column chromatography (pressure: 45 kPa, column: 100mm on length and 16mm on diameter). Ethanol 96% and acetone were compared as eluent. The amount of solvent and yield of curcuminoids used as indicator purification. The silica gel was regenerated with heating in 600°C for 8 hours The silica gels were analyzed by IR spectroscopy and X-ray diffraction. Furthermore, regenerated silica gel was used as the stationary phase in vacuum column chromatography under the same conditions with the previous purification. All the purification experiments were performed in three repetitions. Based on regression equation, y=0.132x+0.0011 (r{sup 2}=0.9997) the yield of curcuminoids on purified products using ethanol as the eluent was improved 4.26% (to 14.3724±0.5749%) and by acetone was improved 3,03% (to 13.1450 ±0.6318%). The IR spectrum of both silica gel showed the same vibration profile and also there were three crystallinity peaks missing on its X-ray diffraction. Regenerated silica gel has the same performance with new silica gel in purification of temulawak’s extract: by ethanol has increased 4.08% (14.1947±0.7415%) and 2.93% (13.0447±0.4822) by acetone. In addition, all purification products showed similar TLC profiles. Purification using regenerated silica gel as the adsorbent on vacuum column chromatography has exactly same potential with the new silica gel.

  9. Fatty acid biosynthesis. VIII. The fate of malonyl-CoA in fatty acid biosynthesis by purified enzymes from lactating-rabbit mammary gland

    DEFF Research Database (Denmark)

    Hansen, Heinz Johs. Max; Carey, E.M.; Dils, R.

    1971-01-01

    - 1. We have investigated the formation and utilization of malonyl-CoA in fatty acid synthesis catalysed by preparations of partially purified acetyl-CoA carboxylase and purified fatty acid synthetase from lactating-rabbit mammary gland. - 2. Carboxylation of [1-14C]acetyl-CoA was linked to fatty...... acid synthesis by the presence of fatty acid synthetase and NADPH. The rate of fatty acid formation was equal to that of acetyl-CoA carboxylation, without the accumulation of free malonyl-CoA to a concentration required to obtain the same rate of fatty acid synthesis from added [1,3-14C2]malonyl......-CoA. - 3. The preparations of acetyl-CoA carboxylase and fatty acid synthetase were each able to decarboxylate [1,3-14C2]malonyl-CoA. - 4. Both enzyme preparations acted as competitive inhibitors of 14CO2 fixation into acetyl-CoA catalysed by acetyl-CoA carboxylase in the absence of NADPH...

  10. The first outbreak of Taenia ovis infection in China.

    Science.gov (United States)

    Shi, Wangui; He, Wei; Guo, Xiaola; Liu, Quanyuan; Gao, Shengzhi; Zhan, Fang; Liu, Xu; Pan, Yonghong; Luo, Xuenong; Zheng, Yadong

    2016-10-01

    Infection of Taenia ovis metacestodes in sheep or goats causes great economic losses due to condemnation of carcasses. T. ovis infection is not formally recorded in China to date. In October, 2015, T. ovis infection occurred in Jingtai County, China, and 113 of 192 sheep from one farm were infected. Cysts resided in the cardiac and skeletal muscle, and evaginated metacestodes had four suckers and scolex armed with approximately 23 hooks. Using cox1 and nad1 as molecular markers, the sample was further identified and the results showed that the cox1 and nad1 nucleotide sequences of the sample shared 99% identity with that of T. ovis and 75%-91.3% with those of other Taenia species. Taken together, these results confirm the first occurrence of T. ovis in China. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  11. Effects of purified lignin on rumen metabolism and growth performance of feedlot cattle

    Directory of Open Access Journals (Sweden)

    Yuxi Wang

    2017-03-01

    Full Text Available Objective The objectives were to assess the effects of purified lignin from wheat straw (sodium hydroxide dehydrated lignin; SHDL on in vitro ruminal fermentation and on the growth performance of feedlot cattle. Methods In vitro experiments were conducted by incubating a timothy-alfalfa (50:50 forage mixture (48 h and barley grain (24 h with 0, 0.25, 0.5, 1.0, and 2.0 mg/mL of rumen fluid (equivalent to 0, 2, 4, 8, and 16 g SHDL/kg diet. Productions of CH4 and total gas, volatile fatty acids, ammonia, dry matter (DM disappearance (DMD and digestion of neutral detergent fiber (NDF or starch were measured. Sixty Hereford-Angus cross weaned steer calves were individually fed a typical barley silage-barley grain based total mixed ration and supplemented with SHDL at 0, 4, 8, and 16 g/kg DM for 70 (growing, 28 (transition, and 121 d (finishing period. Cattle were slaughtered at the end of the experiment and carcass traits were assessed. Results With forage, SHDL linearly (p<0.001 reduced 48-h in vitro DMD from 54.9% to 39.2%, NDF disappearance from 34.1% to 18.6% and the acetate: propionate ratio from 2.56 to 2.41, but linearly (p<0.001 increased CH4 production from 9.5 to 12.4 mL/100 mg DMD. With barley grain, SHDL linearly increased (p<0.001 24-h DMD from74.6% to 84.5%, but linearly (p<0.001 reduced CH4 production from 5.6 to 4.2 mL/100 mg DMD and NH3 accumulation from 9.15 to 4.49 μmol/mL. Supplementation of SHDL did not affect growth, but tended (p = 0.10 to linearly reduce feed intake, and quadratically increased (p = 0.059 feed efficiency during the finishing period. Addition of SHDL also tended (p = 0.098 to linearly increase the saleable meat yield of the carcass from 52.5% to 55.7%. Conclusion Purified lignin used as feed additive has potential to improve feed efficiency for finishing feedlot cattle and carcass quality.

  12. Leukotriene formation by purified 5-lipoxygenase from rat basophilic leukemia cells

    International Nuclear Information System (INIS)

    Hogaboom, G.K.; Cook, M.; Sarau, H.M.; Newton, J.F.; Crooke, S.T.

    1986-01-01

    Arachidonate 5-lipoxygenase (5-LO) from rat basophilic leukemia (RBL-1) cell high speed (105,000 x g for 60 min) supernatants was purified to electrophoretic homogeneity by gel filtration and anion-exchange protein-high pressure liquid chromatography (HPLC). The 5-LO rapidly converted [ 14 C]arachidonate at 20 0 C to [ 14 C]5-hydroperoxyeicosate-traenoic acid (HPETE) as determined by reversed phase-HPLC, scanning spectrophotometry and radiochemical detection. In addition, 5-LO converted both 5-HPETE and arachidonate to 5,12-dihydroxyeicosatetraenoic acids (diHETEs). The 5,12-diHETEs were identified as 6-trans-leukotriene (LT) B4 and 6-trans-12-epi-LTB4 as determined by reversed phase HPLC, scanning spectrophotometry and gas chromatography-mass spectrometry. These data indicate that the RBL-1 5-LO and LTA4 synthetase activities reside on the same protein and that it catalyzes the bioconversion of arachidonate to not only 5-HPETE but also to LTA4. The results suggest that a critical regulatory step in LT biosynthesis in mammalian systems involves the intricate coupling of the enzymes 5-LO and LTA4 synthetase and the interactions of their respective cofactors, substrates and reaction products

  13. Purified reconstituted lac carrier protein from Escherichia coli is fully functional.

    Science.gov (United States)

    Viitanen, P; Garcia, M L; Kaback, H R

    1984-03-01

    Proteoliposomes reconstituted with lac carrier protein purified from the plasma membrane of Escherichia coli catalyze each of the translocation reactions typical of the beta-galactoside transport system (i.e., active transport, counterflow, facilitated influx and efflux) with turnover numbers and apparent Km values comparable to those observed in right-side-out membrane vesicles. Furthermore, detailed kinetic studies show that the reconstituted system exhibits properties analogous to those observed in membrane vesicles. Imposition of a membrane potential (delta psi, interior negative) causes a marked decrease in apparent Km (by a factor of 7 to 10) with a smaller increase in Vmax (approximately equal to 3-fold). At submaximal values of delta psi, the reconstituted carrier exhibits biphasic kinetics, with one component manifesting the kinetic parameters of active transport and the other exhibiting the characteristics of facilitated diffusion. Finally, at low lactose concentrations, the initial velocity of influx varies linearly with the square of the proton electro-chemical gradient. The results provide quantitative support for the contention that a single polypeptide species, the product of the lac y gene, is responsible for each of the transport reactions typical of the beta-galactoside transport system.

  14. Phosphoribosylpyrophosphate synthetase of Escherichia coli. Properties of the purified enzyme and primary structure of the prs gene

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne; Harlow, Kenneth W.; King, Cheryl J.

    1986-01-01

    of ADP. The nucleotide sequence of the E. coli prs gene has been determined and the coding segment established. The deduced amino acid sequence of P-Rib-PP synthetase contained 314 amino acid residues and the molecular weight was calculated as 34,060. The initiation site of transcription was determined......Phosphoribosylpyrophosphate (P-Rib-PP) synthetase of Escherichia coli has been purified to near homogeneity from a strain harboring the prs gene, encoding P-Rib-PP synthetase, on a multicopy plasmid. Analysis of the enzyme showed that it required inorganic phosphate for activity and for stability...

  15. Separation of Am from lanthanides by a synergistic mixture of purified Cyanex 301 and TBP

    International Nuclear Information System (INIS)

    Xinghai Wang; Yongjun Zhu; Rongzhou Jiao

    2002-01-01

    The dependence of the distribution ratios of 241 Am and lanthanides between purified Cyanex 301 (HBTMPDTP)-TBP-kerosene/nitrate solution on pH, lanthanide concentration in aqueous phase and degree of saponification of HBTMPDTP was investigated. The distribution ratios of 241 Am and lanthanides increase with pH and degree of saponification of HBTMPDTP and decrease with lanthanides concentration. Countercurrent multistage extraction consisting of 7 extraction, 3 washing and 2 stripping stages showed that more than 99,99% of 241 Am and less than 0.04% of lanthanides were extracted. The pH 1/2 value of Am was 2.45 compared to 3.16 in case of HBTMPDTP-kerosene extraction. (author)

  16. Protein content and functional characteristics of serum-purified exosomes from patients with colorectal cancer revealed by quantitative proteomics.

    Science.gov (United States)

    Chen, Yanyu; Xie, Yong; Xu, Lai; Zhan, Shaohua; Xiao, Yi; Gao, Yanpan; Wu, Bin; Ge, Wei

    2017-02-15

    Tumor cells of colorectal cancer (CRC) release exosomes into the circulation. These exosomes can mediate communication between cells and affect various tumor-related processes in their target cells. We present a quantitative proteomics analysis of the exosomes purified from serum of patients with CRC and normal volunteers; data are available via ProteomeXchange with identifier PXD003875. We identified 918 proteins with an overlap of 725 Gene IDs in the Exocarta proteins list. Compared with the serum-purified exosomes (SPEs) of normal volunteers, we found 36 proteins upregulated and 22 proteins downregulated in the SPEs of CRC patients. Bioinformatics analysis revealed that upregulated proteins are involved in processes that modulate the pretumorigenic microenvironment for metastasis. In contrast, differentially expressed proteins (DEPs) that play critical roles in tumor growth and cell survival were principally downregulated. Our study demonstrates that SPEs of CRC patients play a pivotal role in promoting the tumor invasiveness, but have minimal influence on putative alterations in tumor survival or proliferation. According to bioinformatics analysis, we speculate that the protein contents of exosomes might be associated with whether they are involved in premetastatic niche establishment or growth and survival of metastatic tumor cells. This information will be helpful in elucidating the pathophysiological functions of tumor-derived exosomes, and aid in the development of CRC diagnostics and therapeutics. © 2016 UICC.

  17. Cerebral cysticercosis in a cat : clinical communication

    Directory of Open Access Journals (Sweden)

    E.V. Schwan

    2002-07-01

    Full Text Available The metacestode of Taenia solium, Cysticercus cellulosae, was recovered from the brain of a cat showing central nervous clinical signs ante mortem. This is the first record of cerebral cysticercosis in a cat in South Africa.

  18. Host-Parasite Interactions and Purifying Selection in a Microsporidian Parasite of Honey Bees.

    Science.gov (United States)

    Huang, Qiang; Chen, Yan Ping; Wang, Rui Wu; Cheng, Shang; Evans, Jay D

    2016-01-01

    To clarify the mechanisms of Nosema ceranae parasitism, we deep-sequenced both honey bee host and parasite mRNAs throughout a complete 6-day infection cycle. By time-series analysis, 1122 parasite genes were significantly differently expressed during the reproduction cycle, clustering into 4 expression patterns. We found reactive mitochondrial oxygen species modulator 1 of the host to be significantly down regulated during the entire infection period. Our data support the hypothesis that apoptosis of honey bee cells was suppressed during infection. We further analyzed genome-wide genetic diversity of this parasite by comparing samples collected from the same site in 2007 and 2013. The number of SNP positions per gene and the proportion of non-synonymous substitutions per gene were significantly reduced over this time period, suggesting purifying selection on the parasite genome and supporting the hypothesis that a subset of N. ceranae strains might be dominating infection.

  19. Taenia saginata cysticercosis in Oyo State, Nigeria | Faleke ...

    African Journals Online (AJOL)

    http://dx.doi.org/10.4314/tv.v22i3.4562 · AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors · FAQ's · More about AJOL · AJOL's Partners · Terms and Conditions of Use · Contact AJOL · News. OTHER RESOURCES... for Researchers · for Journals · for Authors · for Policy Makers ...

  20. The effect of wound instillation of a novel purified capsaicin formulation on postherniotomy pain

    DEFF Research Database (Denmark)

    Aasvang, Eske K; Hansen, Jeanette B; Malmstrøm, Jørgen

    2008-01-01

    , preclinical, and clinical studies, and may be an effective adjunct to postoperative pain management. METHODS: We performed a single-center, randomized, double-blind, placebo-controlled study of the analgesic efficacy of a single intraoperative wound instillation of 1000 microg ultrapurified capsaicin (ALGRX......BACKGROUND: Acute postoperative pain is common after most surgical procedures. Despite the availability of many analgesic options, postoperative pain management is often unsatisfactory. Purified capsaicin (ALGRX 4975 98% pure) has demonstrated prolong inhibition of C-fiber function in in vitro...... 4975) after open mesh groin hernia repair in 41 adult male patients. The primary end-point was average daily visual analog scale (VAS) pain scores during the first week after surgery assessed as area under the curve (AUC). Pain was recorded twice daily in a pain diary for 4 wk. Physical examination...

  1. Human Pol ζ purified with accessory subunits is active in translesion DNA synthesis and complements Pol η in cisplatin bypass.

    Science.gov (United States)

    Lee, Young-Sam; Gregory, Mark T; Yang, Wei

    2014-02-25

    DNA polymerase ζ (Pol ζ) is a eukaryotic B-family DNA polymerase that specializes in translesion synthesis and is essential for normal embryogenesis. At a minimum, Pol ζ consists of a catalytic subunit Rev3 and an accessory subunit Rev7. Mammalian Rev3 contains >3,000 residues and is twice as large as the yeast homolog. To date, no vertebrate Pol ζ has been purified for biochemical characterization. Here we report purification of a series of human Rev3 deletion constructs expressed in HEK293 cells and identification of a minimally catalytically active human Pol ζ variant. With a tagged form of an active Pol ζ variant, we isolated two additional accessory subunits of human Pol ζ, PolD2 and PolD3. The purified four-subunit Pol ζ4 (Rev3-Rev7-PolD2-PolD3) is much more efficient and more processive at bypassing a 1,2-intrastrand d(GpG)-cisplatin cross-link than the two-subunit Pol ζ2 (Rev3-Rev7). We show that complete bypass of cisplatin lesions requires Pol η to insert dCTP opposite the 3' guanine and Pol ζ4 to extend the primers.

  2. Inhibition of purified enolases from oral bacteria by fluoride.

    Science.gov (United States)

    Guha-Chowdhury, N; Clark, A G; Sissons, C H

    1997-04-01

    Enolase activity in strains of oral streptococci previously has been found to be inhibited by 50% (Ki) by fluoride concentrations ranging from 50 to 300 microM or more in the presence of 0.5 to 1.0 mM inorganic phosphate ions. In this study, enolase was extracted and partly purified by a two-step process from five oral bacterial species and the effect of fluoride on the kinetics of enolase examined. The molecular weight of the putative enolase proteins was 46-48 kDa. The Vmax values ranged from 20 to 323 IU/mg and K(m) for glycerate-2-phosphate from 0.22 to 0.74 mM. Enolase activity was inhibited competitively by fluoride, with Ki values ranging from 16 to 54 microM in the presence of 5 mM inorganic phosphate ions. Ki values for phosphate ranged from 2 to 8 mM. The enolase from Streptococcus sanguis ATCC 10556 was more sensitive to fluoride (Ki = 16 +/- 2) than was enolase from Streptococcus salivarius ATCC 10575 (Ki = 19 +/- 2) or Streptococcus mutans NCTC 10449 (Ki = 40 +/- 4) and all three streptococcal strains were more sensitive to fluoride than either Actinomyces naeslundii WVU 627 (Ki = 46 +/- 6) or Lactobacillus rhamnosus ATCC 7469 (Ki = 54 +/- 6) enolases. The levels of fluoride found to inhibit the streptococcal enolases in this study are much lower than previously reported and are likely to be present in plaque, especially during acidogenesis, and could exert an anti-glycolytic effect.

  3. Selective Biological Responses of Phagocytes and Lungs to Purified Histones.

    Science.gov (United States)

    Fattahi, Fatemeh; Grailer, Jamison J; Lu, Hope; Dick, Rachel S; Parlett, Michella; Zetoune, Firas S; Nuñez, Gabriel; Ward, Peter A

    2017-01-01

    Histones invoke strong proinflammatory responses in many different organs and cells. We assessed biological responses to purified or recombinant histones, using human and murine phagocytes and mouse lungs. H1 had the strongest ability in vitro to induce cell swelling independent of requirements for toll-like receptors (TLRs) 2 or 4. These responses were also associated with lactate dehydrogenase release. H3 and H2B were the strongest inducers of [Ca2+]i elevations in phagocytes. Cytokine and chemokine release from mouse and human phagocytes was predominately a function of H2A and H2B. Double TLR2 and TLR4 knockout (KO) mice had dramatically reduced cytokine release induced in macrophages exposed to individual histones. In contrast, macrophages from single TLR-KO mice showed few inhibitory effects on cytokine production. Using the NLRP3 inflammasome protocol, release of mature IL-1β was predominantly a feature of H1. Acute lung injury following the airway delivery of histones suggested that H1, H2A, and H2B were linked to alveolar leak of albumin and the buildup of polymorphonuclear neutrophils as well as the release of chemokines and cytokines into bronchoalveolar fluids. These results demonstrate distinct biological roles for individual histones in the context of inflammation biology and the requirement of both TLR2 and TLR4. © 2017 S. Karger AG, Basel.

  4. Structure-Guided Design of an Engineered Streptavidin with Reusability to Purify Streptavidin-Binding Peptide Tagged Proteins or Biotinylated Proteins

    OpenAIRE

    Wu, Sau-Ching; Wong, Sui-Lam

    2013-01-01

    Development of a high-affinity streptavidin-binding peptide (SBP) tag allows the tagged recombinant proteins to be affinity purified using the streptavidin matrix without the need of biotinylation. The major limitation of this powerful technology is the requirement to use biotin to elute the SBP-tagged proteins from the streptavidin matrix. Tight biotin binding by streptavidin essentially allows the matrix to be used only once. To address this problem, differences in interactions of biotin an...

  5. PRESERVATIVE POTENTIAL OF PURIFIED BACTERIOCIN PRODUCED FROM BREVIBACILLUS BORSTELENSIS AG1 ISOLATED FROM MARCHA – A TRADITIONAL WINE STARTER CULTURE CAKE IN TOMATO PASTE

    Directory of Open Access Journals (Sweden)

    Anupama Gupta

    2015-04-01

    Full Text Available Purified bacteriocin produced from Brevibacillus borstelensis AG1 isolated from Marcha a local wine starter herbal cake, was used to enhance the shelf life of tomato paste. Preservative effect of purified bacteriocin was studied for nine days in tomato paste inoculated with food borne pathogens and was compared to commercial biopreservative – nisin and chemical preservative – sodium benzoate. The indicator strains i.e. Listeria monocytogenes MTCC839, Bacillus subtilis CRI and Clostridium perfringens MTCC1739 were used at the amount 8.16, 8.13 and 8.18 log CFU/ml. Viable cells were counted periodically and a consistent reduction in number of viable cells of each tested pathogen was observed. It was found antagonistic against L. monocytogenes MTCC839, B. subtilis CRI and C. perfringes MTCC1739 which are the most challengeable and food borne pathogens found in processed vegetables products. Purified bacteriocin was found active over a wide pH range i.e. 3.0 to 11.0 and was able to withstand temperature up to 100oC. It showed a better preservative potential by reducing pathogenic load of the tested strains (by 2.02, 2.05 and 2.02 log cycles (CFU/ml of L. monocytogenes MTCC839, B. subtilis CRI and C. perfringes MTCC1739, respectively in tomato paste as compared to control (without bacteriocin. This proves efficiency of bacteriocin produced by B. borstelensis AG1 as biopreservative to enhance the safety and shelf life of acidic foods.

  6. Purifying, Separating, and Concentrating Cells From a Sample Low in Biomass

    Science.gov (United States)

    Benardini, James N.; LaDuc, Myron T.; Diamond, Rochelle

    2012-01-01

    Frequently there is an inability to process and analyze samples of low biomass due to limiting amounts of relevant biomaterial in the sample. Furthermore, molecular biological protocols geared towards increasing the density of recovered cells and biomolecules of interest, by their very nature, also concentrate unwanted inhibitory humic acids and other particulates that have an adversarial effect on downstream analysis. A novel and robust fluorescence-activated cell-sorting (FACS)-based technology has been developed for purifying (removing cells from sampling matrices), separating (based on size, density, morphology), and concentrating cells (spores, prokaryotic, eukaryotic) from a sample low in biomass. The technology capitalizes on fluorescent cell-sorting technologies to purify and concentrate bacterial cells from a low-biomass, high-volume sample. Over the past decade, cell-sorting detection systems have undergone enhancements and increased sensitivity, making bacterial cell sorting a feasible concept. Although there are many unknown limitations with regard to the applicability of this technology to environmental samples (smaller cells, few cells, mixed populations), dogmatic principles support the theoretical effectiveness of this technique upon thorough testing and proper optimization. Furthermore, the pilot study from which this report is based proved effective and demonstrated this technology capable of sorting and concentrating bacterial endospore and bacterial cells of varying size and morphology. Two commercial off-the-shelf bacterial counting kits were used to optimize a bacterial stain/dye FACS protocol. A LIVE/DEAD BacLight Viability and Counting Kit was used to distinguish between the live and dead cells. A Bacterial Counting Kit comprising SYTO BC (mixture of SYTO dyes) was employed as a broad-spectrum bacterial counting agent. Optimization using epifluorescence microscopy was performed with these two dye/stains. This refined protocol was further

  7. Analysis of purified gp96 preparations from rat and mouse livers using 2-D gel electrophoresis and tandem mass spectrometry.

    Science.gov (United States)

    Fairburn, B; Muthana, M; Hopkinson, K; Slack, L K; Mirza, S; Georgiou, A S; Espigares, E; Wong, C; Pockley, A G

    2006-09-01

    The stress protein gp96 exhibits a number of immunological activities, the majority of studies into which have used gp96 purified from a variety of tissues. On the basis of 1-D gel electrophoresis, the purity of these preparations has been reported to range between 70% and 99%. This study analyzed gp96 preparations from rat and mouse livers using 2-D gel electrophoresis and liquid chromatography electrospray ionization tandem mass spectrometry (MS-MS). The procedure for purifying gp96 was reproducible, as similar protein profiles were observed in replicate gels of gp96 preparations. The purity of the preparations was typically around 70%, with minor co-purified proteins of varying molecular weights and mobilities being present. Dominant bands at 95-100 kDa in preparations from Wistar rats and C57BL/6 mice were identified as gp96 by ECL Western blotting. Multiple bands having similar, yet distinct molecular weights and differing pI mobility on ECL Western blots were confirmed as being gp96 in preparations from Wistar rats using MS-MS. The most striking feature of the 2-D gel analysis was the presence of additional dominant bands at 55 kDa in preparations from Wistar rats, and at 75-90 kDa in preparations from C57BL/6 mice. These were identified as gp96 by ECL Western blotting and, in the case of preparations from Wistar rats, by MS-MS. Although the lower molecular weight, gp96-related molecules might be partially degraded gp96, their reproducible presence, definition and characteristics suggest that they are alternative, species-specific isoforms of the molecule. A 55 kDa protein which exhibited a lower pI value than gp96 was present in all preparations and this was identified as calreticulin, another putative immunoregulatory molecule. This study confirms the reproducibility of the gp96 purification protocol and reveals the presence of multiple gp96 isoforms, some of which likely result from post-translational modifications such as differential glycosylation and

  8. Uso del aralen en el tratamiento de la teniasis

    Directory of Open Access Journals (Sweden)

    Carlos Arteaga Camargo

    1951-08-01

    Full Text Available En el presente trabajo se hace referencia al tratamiento de siete casos de Taeniasis (todos de Taenia saginata, por medio del "Aralen". Dicho tratamiento es de administración fácil, no produjo manifestaciones tóxicas y dio éxito en todos los casos tratados.

  9. Efficacy of soluble glycoprotein fraction from Allium sativum purified by size exclusion chromatography on murine Schistosomiasis mansoni.

    Science.gov (United States)

    Aly, Ibrahim; Taher, Eman E; El-Sayed, Hoda; Mohammed, Faten A; ELnain, Gehan; Hamad, Rabab S; Bayoumy, Elsayed M

    2017-06-01

    In this work, the efficiency of crude MeOH extracts and soluble glycoprotein fraction of Allium sativum purified by size-exclusion chromatography (SEC) on parasitological, histopathological and some biochemical parameters in Schistosoma mansoni infected mice were investigated. Animals were infected by tail immersion with 100 cercariae/each mouse and divided into five groups in addition to the normal control. The results revealed a significant decrease in mean worm burden in all treated mice especially in the group treated with soluble glycoprotein fraction of A. sativum as compared to infected non-treated control with the disappearance of female worms. Administration of the studied extracts revealed remarkable amelioration in the levels of all the measured parameters in S. mansoni infected mice. In addition, treatment of mice with crude A. sativum MeOH extract and soluble glycoprotein fraction of A. sativum decreased significantly the activities of studied enzymes as compared to the infected untreated group. The highest degrees of enhancement in pathological changes was observed in the treated one with soluble glycoprotein fraction of A. sativum compared to the infected group represented by small sized, late fibro-cellular granuloma, the decrease in cellular constituents and degenerative changes in eggs. In conclusion, A. sativum treatment had effective schistosomicidal activities, through reduction of worm burden and tissue eggs, especially when it was given in purified glycoprotein fraction. Moreover, the soluble glycoprotein fraction of A. sativum largely modulates both the size and the number of granulomas. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Integration-deficient lentivectors: an effective strategy to purify and differentiate human embryonic stem cell-derived hepatic progenitors.

    Science.gov (United States)

    Yang, Guanghua; Si-Tayeb, Karim; Corbineau, Sébastien; Vernet, Rémi; Gayon, Régis; Dianat, Noushin; Martinet, Clémence; Clay, Denis; Goulinet-Mainot, Sylvie; Tachdjian, Gérard; Tachdjian, Gérard; Burks, Deborah; Vallier, Ludovic; Bouillé, Pascale; Dubart-Kupperschmitt, Anne; Weber, Anne

    2013-07-19

    Human pluripotent stem cells (hPSCs) hold great promise for applications in regenerative medicine. However, the safety of cell therapy using differentiated hPSC derivatives must be improved through methods that will permit the transplantation of homogenous populations of a specific cell type. To date, purification of progenitors and mature cells generated from either embryonic or induced pluripotent stem cells remains challenging with use of conventional methods. We used lentivectors encoding green fluorescent protein (GFP) driven by the liver-specific apoliprotein A-II (APOA-II) promoter to purify human hepatic progenitors. We evaluated both integrating and integration-defective lentivectors in combination with an HIV integrase inhibitor. A human embryonic stem cell line was differentiated into hepatic progenitors using a chemically defined protocol. Subsequently, cells were transduced and sorted at day 16 of differentiation to obtain a cell population enriched in hepatic progenitor cells. After sorting, more than 99% of these APOA-II-GFP-positive cells expressed hepatoblast markers such as α-fetoprotein and cytokeratin 19. When further cultured for 16 days, these cells underwent differentiation into more mature cells and exhibited hepatocyte properties such as albumin secretion. Moreover, they were devoid of vector DNA integration. We have developed an effective strategy to purify human hepatic cells from cultures of differentiating hPSCs, producing a novel tool that could be used not only for cell therapy but also for in vitro applications such as drug screening. The present strategy should also be suitable for the purification of a broad range of cell types derived from either pluripotent or adult stem cells.

  11. Population genetic evidence for positive and purifying selection acting at the human IFN-γ locus in Africa.

    Science.gov (United States)

    Campbell, Michael C; Smith, Lunden T; Harvey, Jayla

    2018-03-29

    Despite its critical role in the defense against microbial infection and tumor development, little is known about the range of nucleotide and haplotype variation at IFN-γ, or the evolutionary forces that have shaped patterns of diversity at this locus. To address this gap in knowledge, we examined sequence data from the IFN-γ gene in 1461 individuals from 15 worldwide populations. Our analyses uncovered novel patterns of variation in distinct African populations, including an excess of high frequency-derived alleles, unusually long haplotype structure surrounding the IFN-γ gene, and a "star-like" genealogy of African-specific haplotypes carrying variants previously associated with infectious disease. We also inferred a deep time to coalescence of variation at IFN-γ (~ 0.8 million years ago) and ancient ages for common polymorphisms predating the evolution of modern humans. Taken together, these results are congruent with a model of positive selection on standing variation in African populations. Furthermore, we inferred that common variants in intron 3 of IFN-γ are the likely targets of selection. In addition, we observed a paucity of non-synonymous substitutions relative to synonymous changes in the exons of IFN-γ in African and non-African populations, suggestive of strong purifying selection. Therefore, we contend that positive and purifying selection have influenced levels of diversity in different regions of IFN-γ, implying that these distinct genic regions are, or have been, functionally important. Overall, this study provides additional insights into the evolutionary events that have contributed to the frequency and distribution of alleles having a role in human health and disease.

  12. Host-Parasite Interactions and Purifying Selection in a Microsporidian Parasite of Honey Bees.

    Directory of Open Access Journals (Sweden)

    Qiang Huang

    Full Text Available To clarify the mechanisms of Nosema ceranae parasitism, we deep-sequenced both honey bee host and parasite mRNAs throughout a complete 6-day infection cycle. By time-series analysis, 1122 parasite genes were significantly differently expressed during the reproduction cycle, clustering into 4 expression patterns. We found reactive mitochondrial oxygen species modulator 1 of the host to be significantly down regulated during the entire infection period. Our data support the hypothesis that apoptosis of honey bee cells was suppressed during infection. We further analyzed genome-wide genetic diversity of this parasite by comparing samples collected from the same site in 2007 and 2013. The number of SNP positions per gene and the proportion of non-synonymous substitutions per gene were significantly reduced over this time period, suggesting purifying selection on the parasite genome and supporting the hypothesis that a subset of N. ceranae strains might be dominating infection.

  13. An analysis of the repair processes in ultraviolet-irradiated Micrococcus luteus using purified ultraviolet-endonuclease

    International Nuclear Information System (INIS)

    Tomilin, N.V.; Zherebtsov, S.V.

    1982-01-01

    The measurement of the frequency of endonucleolytic incisions in ultraviolet-irradiated DNA serves as the test for the presence of pyrimidine dimers. In accordance with this approach, the lysates of three Micrococcus luteus strains containing radioactively labeled chromosomes were treated with purified M. luteus ultraviolet-endonuclease to trace segregation of dimers amongst parental and newly synthesized DNA and their removal during postreplication and excision DNA repair. A considerable proportion of the dimers in all strains tested proved to be insensitive to the action of exogenous incising enzyme. The use of chloramphenicol as an inhibitor of postirradiation protein synthesis in combination with ultraviolet-endonuclease treatment of DNA allowed to reveal at least two alternative pathways of postreplication repair: constitutively active recombinational pathway and inducible nonrecombinational one. (Auth.)

  14. Passive administration of purified secretory IgA from human colostrum induces protection against Mycobacterium tuberculosis in a murine model of progressive pulmonary infection

    Directory of Open Access Journals (Sweden)

    Alvarez Nadine

    2013-02-01

    Full Text Available Abstract Background Immunoglobulin A is the most abundant isotype in secretions from mucosal surfaces of the gastrointestinal, respiratory and genitourinary tracts and in external secretions such as colostrum, breast milk, tears and saliva. The high concentration of human secretory IgA (hsIgA in human colostrum strongly suggests that it should play an important role in the passive immune protection against gastrointestinal and respiratory infections. Materials and methods Human secretory IgA was purified from colostrum. The reactivity of hsIgA against mycobacterial antigens and its protective capacity against mycobacterial infection was evaluated. Results The passive administration of hsIgA reduces the pneumonic area before challenge with M. tuberculosis. The intratracheal administration of M. tuberculosis preincubated with hsIgA to mice greatly reduced the bacterial load in the lungs and diminished lung tissue injury. Conclusions HsIgA purified from colostrum protects against M. tuberculosis infection in an experimental mouse model.

  15. Instantaneous Purified Orbit: A New Tool for Analysis of Nonstationary Vibration of Rotor System

    Directory of Open Access Journals (Sweden)

    Shi Dongfeng

    2001-01-01

    Full Text Available In some circumstances, vibration signals of large rotating machinery possess time-varying characteristics to some extent. Traditional diagnosis methods, such as FFT spectrum and orbit diagram, are confronted with a huge challenge to deal with this problem. This work aims at studying the four intrinsic drawbacks of conventional vibration signal processing method and instantaneous purified orbit (IPO on the basis of improved Fourier spectrum (IFS to analyze nonstationary vibration. On account of integration, the benefits of short period Fourier transform (SPFT and regular holospectrum, this method can intuitively reflect vibration characteristics of’a rotor system by means of parameter analysis for corresponding frequency ellipses. Practical examples, such as transient vibration in run-up stages and bistable condition of rotor show that IPO is a powerful tool for diagnosis and analysis of the vibration behavior of rotor systems.

  16. Enriched surface acidity for surfactant-free suspensions of carboxylated carbon nanotubes purified by centrifugation

    Directory of Open Access Journals (Sweden)

    Elizabeth I. Braun

    2016-06-01

    Full Text Available It is well known that surfactant-suspended carbon nanotube (CNT samples can be purified by centrifugation to decrease agglomerates and increase individually-dispersed CNTs. However, centrifugation is not always part of protocols to prepare CNT samples used in biomedical applications. Herein, using carboxylated multi-walled CNTs (cMWCNTs suspended in water without a surfactant, we developed a Boehm titrimetric method for the analysis of centrifuged cMWCNT suspensions and used it to show that the surface acidity of oxidized carbon materials in aqueous cMWCNT suspensions was enriched by ∼40% by a single low-speed centrifugation step. This significant difference in surface acidity between un-centrifuged and centrifuged cMWCNT suspensions has not been previously appreciated and is important because the degree of surface acidity is known to affect the interactions of cMWCNTs with biological systems.

  17. Retinoblastoma protein co-purifies with proteasomal insulin-degrading enzyme: Implications for cell proliferation control

    Energy Technology Data Exchange (ETDEWEB)

    Radulescu, Razvan T., E-mail: ratura@gmx.net [Molecular Concepts Research (MCR), Muenster (Germany); Duckworth, William C. [Department of Medicine, Phoenix VA Health Care System, Phoenix, AZ (United States); Levy, Jennifer L. [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States); Fawcett, Janet, E-mail: janet.fawcett@va.gov [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States)

    2010-04-30

    Previous investigations on proteasomal preparations containing insulin-degrading enzyme (IDE; EC 3.4.24.56) have invariably yielded a co-purifying protein with a molecular weight of about 110 kDa. We have now found both in MCF-7 breast cancer and HepG2 hepatoma cells that this associated molecule is the retinoblastoma tumor suppressor protein (RB). Interestingly, the amount of RB in this protein complex seemed to be lower in HepG2 vs. MCF-7 cells, indicating a higher (cytoplasmic) protein turnover in the former vs. the latter cells. Moreover, immunofluorescence showed increased nuclear localization of RB in HepG2 vs. MCF-7 cells. Beyond these subtle differences between these distinct tumor cell types, our present study more generally suggests an interplay between RB and IDE within the proteasome that may have important growth-regulatory consequences.

  18. Method and apparatus for purifying nucleic acids and performing polymerase chain reaction assays using an immiscible fluid

    Energy Technology Data Exchange (ETDEWEB)

    Koh, Chung-Yan; Light, Yooli Kim; Piccini, Matthew Ernest; Singh, Anup K.

    2017-10-31

    Embodiments of the present invention are directed toward devices, systems, and methods for purifying nucleic acids to conduct polymerase chain reaction (PCR) assays. In one example, a method includes generating complexes of silica beads and nucleic acids in a lysis buffer, transporting the complexes through an immiscible fluid to remove interfering compounds from the complexes, further transporting the complexes into a density medium containing components required for PCR where the nucleic acids disassociate from the silica beads, and thermocycling the contents of the density medium to achieve PCR. Signal may be detected from labeling agents in the components required for PCR.

  19. Effects on the development of Dipylidium caninum and on the host reaction to this parasite in the adult flea (Ctenocephalides felis felis).

    Science.gov (United States)

    Pugh, R E

    1987-01-01

    Temperature was found to be a major factor affecting the development of Dipylidium caninum and the presence of a host reaction of adult Ctenocephalides felis felis to D. caninum. Adult fleas reared at 30-32 degrees C contained fully developed metacestodes when they emerged from their cocoons. However at lower temperatures, D. caninum could not complete development until the flea hosts had spent some time on their mammalian hosts. It was the surface temperature of the mammals (31-36 degrees C) and not the fleas' blood meals which resulted in the metacestodes completing their development. This development of D. caninum was therefore independent of the flea development. At 20 degrees C, a larger and more prolonged host reaction was mounted than at higher temperatures. The larval flea diet had a small effect on the subsequent cestode development and the adult fleas' reaction to it.

  20. Development of a Microwave Regenerative Sorbent-Based Hydrogen Purifier

    Science.gov (United States)

    Wheeler, Richard R., Jr.; Dewberry, Ross H.; McCurry, Bryan D.; Abney, Morgan B.; Greenwood, Zachary W.

    2016-01-01

    This paper describes the design and fabrication of a Microwave Regenerative Sorbent-based Hydrogen Purifier (MRSHP). This unique microwave powered technology was developed for the purification of a hydrogen stream produced by the Plasma Pyrolysis Assembly (PPA). The PPA is a hydrogen recovery (from methane) post processor for NASA's Sabatier-based carbon dioxide reduction process. Embodied in the Carbon dioxide Reduction Assembly (CRA), currently aboard the International Space Station (ISS), the Sabatier reaction employs hydrogen to catalytically recover oxygen, in the form of water, from respiratory carbon dioxide produced by the crew. This same approach is base-lined for future service in the Air Revitalization system on extended missions into deep space where resupply is not practical. Accordingly, manned exploration to Mars may only become feasible with further closure of the air loop as afforded by the greater hydrogen recovery permitted by the PPA with subsequent hydrogen purification. By utilizing the well-known high sorbate loading capacity of molecular sieve 13x, coupled with microwave dielectric heating phenomenon, MRSHP technology is employed as a regenerative filter for a contaminated hydrogen gas stream. By design, freshly regenerated molecular sieve 13x contained in the MRSHP will remove contaminants from the effluent of a 1-CM scale PPA for several hours prior to breakthrough. By reversing flow and pulling a relative vacuum the MRSHP prototype then uses 2.45 GHz microwave power, applied through a novel coaxial antenna array, to rapidly heat the sorbent bed and drive off the contaminants in a short duration vacuum/thermal contaminant desorption step. Finally, following rapid cooling via room temperature cold plates, the MRSHP is again ready to serve as a hydrogen filter.

  1. Epidemiology of taeniosis/cysticercosis in Europe, a systematic review:Western Europe

    OpenAIRE

    Laranjo-González, Minerva; Devleesschauwer, Brecht; Trevisan, Chiara; Allepuz, Alberto; Sotiraki, Smaragda; Abraham, Annette; Afonso, Mariana Boaventura; Blocher, Joachim; Cardoso, Luís; Correia Da Costa, José Manuel; Dorny, Pierre; Gabriël, Sarah; Gomes, Jacinto; Gómez-Morales, María Ángeles; Jokelainen, Pikka

    2017-01-01

    Abstract Background Taenia solium and Taenia saginata are zoonotic parasites of public health importance. Data on their occurrence in humans and animals in western Europe are incomplete and fragmented. In this study, we aimed to update the current knowledge on the epidemiology of these parasites in this region. Methods We conducted a ...

  2. Purification by ozonolysis of 18O enriched water after cyclotron irradiation and the utilization of the purified water for the production of [18F]-FDG (2-deoxy-2-[18F]-fluoro-D-glucose)

    International Nuclear Information System (INIS)

    Asti, M.; Grassi, E.; Sghedoni, R.; De Pietri, G.; Fioroni, F.; Versari, A.; Borasi, G.; Salvo, D.

    2007-01-01

    The high cost of virgin 18 O-enriched water has forced many researchers to study methods to purify and recycle enriched water after the first irradiation for the production of radiopharmaceuticals. In our study, [ 18 O]H 2 O was purified by ozonolysis and distillation. Analyses showed a large decrease in impurities after this treatment. The purification procedure was carried out after the production of 94 batches of [ 18 F]-FDG, which were manufactured using a GE Minitrace cyclotron and a GE Mx TracerLab synthesizer. Saturation yields after bombardment, using virgin and re-purified water were, respectively, 2864±204 MBq/μA and 2727±167 MBq/μA, a decrease of 5.5%. The decrease in [ 18 F]-FDG yield, from 67.2±0.7% to 65.5±0.9%, can be ascribed to the irradiation step only

  3. Synthèses enzymatiques de néoglucoconjugués catalysées par l'alpha-glucosidase purifiée de la blatte Periplaneta americana (Linnaeus

    Directory of Open Access Journals (Sweden)

    Kamenan A.

    2005-01-01

    Full Text Available Enzymatic synthesis of neoglucoconjugates by purified α-glucosidase from cockroach Periplaneta americana (Linnaeus. Cockroach Periplaneta americana (Linnaeus contains in his digestive tract an acid (pH 5,0 and mesophile (50°C α-glucosidase. This enzyme, purified to homogeneity, hydrolyses highly maltose, sucrose and p-nitrophenyl-α-Dglucopyranoside. The ability of α-glucosidase from cockroach purified to homogeneity to catalyse transglucosylation reactions was tested using maltose and saccharose as glucosyl donors and 2-phenylethanol and phenol as acceptors. The experimental conditions were optimized in relation to the time course of the reaction, pH and concentrations of glucosyl donors and acceptors. The yields in transglucosylation reactions at 37 °C were very high and could attain 67% and 48% with 2-phenylethanol and phenol respectively as glucosyl acceptors. This α-glucosidase hydrolyzed the products formed. It seems that the products formed were the phenylethyl-α-D-glucoside and phenyl-α-D-glucoside. These results suggest that α- glucosidase from cockroach is an exoglucosidase which catalyse the splitting of the α-glucosyl residue from the non reducing terminal of the substrate to liberate α-glucose. This comportment indicates that this enzyme operated by a mechanism involving the retention of the anomeric configuration. On the basis of this work, α-glucosidase from P. americana appears to be a valuable tool for the preparation of α-neoglucoconjugates.

  4. Behavioral and hormonal changes associated with the infective dose in experimental taeniasis in golden hamsters (Mesocricetus auratus).

    Science.gov (United States)

    Domínguez-Roldan, Rosa; Hallal-Calleros, Claudia; Sciutto, Edda; Hernández, Marisela; Aguirre-Flores, Virginio; García-Jiménez, Sara; Báez-Saldaña, Armida; Flores-Pérez, Fernando Iván

    2016-07-01

    It has been reported that behavioral changes relate to infection in different parasitoses. However, the relation between the extent of the behavioral changes and the magnitude of the infection has been scarcely studied. The aim of this study was to evaluate the relationship between different doses of infection and the behavioral changes induced in the experimental Taenia pisiformis taeniasis in golden hamsters. Groups of nine hamsters were infected with three or six T. pisiformis metacestodes. The locomotor activity was quantified daily in an open field test during the 21 days after infection; anxiety test was performed in an elevated plus-maze with a dark/light area at 7, 14 and 21 days post-infection, and serum cortisol levels were determined by radioimmunoassay before infection and at day 22 after infection. The challenge itself induced modifications on behavior and cortisol levels in hamsters, with or without successful infection (taenia development). Animals challenged with three metacestodes induced a decrease in locomotor activity and an increase in anxiety in infected animals. A higher and earlier decrease in locomotor activity and increased anxiety levels were observed in hamsters challenged with six cysticerci, which were accompanied by higher levels of sera cortisol at the end of the experiment. At necropsy, 44-55% of hamster became infected with an efficiency of implantation of 22-26%, challenged with three or six cysticerci respectively. The challenge of hamsters with metacestodes, promote behavioral changes in an extent dependent on the magnitude of the challenge, disregarding the effectiveness of the infection. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Effectiveness of purified sulfur intake per os to reduce a reaction in radiotherapy of uterine cervix

    International Nuclear Information System (INIS)

    Smirnova, O.V.; Saliev, V.P.; Klemparskaya, N.N.; Dobronravova, N.N.

    1991-01-01

    Theoretical basis of this work is the development of autosensitization in exposure to ionizing radiation and well-known desensitizing action of sulfuric agents. To reduce clinical manifestations of a side reaction to combined radiotherapy 34 women with diagnosis of cervical cancer were given 0.5-1 g of purified sulfur mixed with 0.25 g of glucose in the morning every 2-3 hours before irradiation, per os; 24 patients received placebo, in 21 patients no protective means were used. All 79 patients were given unified adjuvant therapy and diet No.15. A significant decrease in the reaction to therapeutic irradiation was noted in the study group. No side effects were observed

  6. Phosphoribosylpyrophosphate synthetase of Escherichia coli. Properties of the purified enzyme and primary structure of the prs gene

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne; Harlow, Kenneth W.; King, Cheryl J.

    1986-01-01

    Phosphoribosylpyrophosphate (P-Rib-PP) synthetase of Escherichia coli has been purified to near homogeneity from a strain harboring the prs gene, encoding P-Rib-PP synthetase, on a multicopy plasmid. Analysis of the enzyme showed that it required inorganic phosphate for activity and for stability...... the UAA translation stop codon, within a Thy-rich region following an inverted repeat sequence, indicative of an rho-independent transcription terminator....

  7. Carboxyl group modification significantly altered the kinetic properties of purified carboxymethylcellulase from Aspergillus niger.

    Science.gov (United States)

    Siddiqui; Saqib; Rashid; Rajoka

    2000-10-01

    Carboxymethylcellulase (CMCase) from Aspergillus niger NIAB280 was purified by a combination of ammonium sulphate precipitation, ion-exchange, hydrophobic interaction and gel filtration chromatography on FPLC with 9-folds increase in specific activity. Native and subunit molecular weights were found to be 36 kDa each. The purified CMCase was modified by 1-ethyl-3(3-dimethylaminopropyl) carbodiimide (EDC) in the presence of glycinamide for 15 min (GAM15) and glycinamide plus cellobiose for 75 min (GAM75). Similarly, the enzyme was modified by EDC in the presence of ethylenediamine dihydrochloride plus cellobiose for 75 min (EDAM75). The neutralization (GAM15 and GAM75) and reversal (EDAM75) of negative charges of carboxyl groups of CMCase had profound effect on the specificity constant (k(cat)/K(m)), pH optima, pK(a)'s of the active-site residues and thermodynamic parameters of activation. The specificity constants of native, GAM15, GAM75, and EDAM75 were 143, 340, 804, and 48, respectively. The enthalpy of activation (DeltaH(#)) of Carboxymethylcellulose (CMC) hydrolysis of native (50 and 15 kJ mol(-1)) and GAM15 (41 and 16 kJ mol(-1)) were biphasic whereas those of GAM75 (43 kJ mol(-1)) and EDAM75 (41 k J mol(-1)) were monophasic. Similarly, the entropy of activation (DeltaS(#)) of CMC hydrolysis of native (-61 and -173 J mol(-1) K(-1)) and GAM15 (-91 and -171 J mol(-1) K(-1)) were biphasic whereas those of GAM75 (-82 J mol(-1) K(-1)) and EDAM75 (-106 J mol(-1) K(-1)) were monophasic. The pH optima/pK(a)'s of both acidic and basic limbs of charge neutralized CMCases increased compared with those of native enzyme. The CMCase modification in the presence of glycinamide and absence of cellobiose at different pH's periodically activated and inhibited the enzyme activity indicating conformational changes. We believe that the alteration of the surface charges resulted in gross movement of loops that surround the catalytic pocket, thereby inducing changes in the vicinity

  8. MANUSCRIPT NUMBER: NVJ/10/02/08

    African Journals Online (AJOL)

    SALIS LAWAN

    zoonotic disease caused by the metacestode (hydatid cyst) stage of the dog tapeworm. Echinicoccus granulosus .... TABLE III: Prevalence of hydatid cysts infected organs at Damatura abattoir, 2006. Species. No. of animals examined at slaughter. No. (%) of organs infected. Lungs Liver. Spleen. Sheep. Goats. Total. 3672.

  9. Lipofection of purified adeno-associated virus Rep68 protein: toward a chromosome-targeting nonviral particle.

    Science.gov (United States)

    Lamartina, S; Roscilli, G; Rinaudo, D; Delmastro, P; Toniatti, C

    1998-09-01

    Adeno-associated virus (AAV) integrates very efficiently into a specific site (AAVS1) of human chromosome 19. Two elements of the AAV genome are sufficient: the inverted terminal repeats (ITRs) and the Rep78 or Rep68 protein. The incorporation of the AAV integration machinery in nonviral delivery systems is of great interest for gene therapy. We demonstrate that purified recombinant Rep68 protein is functionally active when directly delivered into human cells by using the polycationic liposome Lipofectamine, promoting the rescue-replication of a codelivered ITR-flanked cassette in adenovirus-infected cells and its site-specific integration in noninfected cells. The sequencing of cloned virus-host DNA junctions confirmed that lipofected Rep68 protein triggers site-specific integration at the same sites in chromosome 19 already characterized in cells latently infected with AAV.

  10. Modulation of phenotypic and functional maturation of murine dendritic cells (DCs) by purified Achyranthes bidentata polysaccharide (ABP).

    Science.gov (United States)

    Zou, Yaxuan; Meng, Jingjuan; Chen, Wenna; Liu, Jingling; Li, Xuan; Li, Weiwei; Lu, Changlong; Shan, Fengping

    2011-08-01

    There are a large number of interactions at molecular and cellular levels between the plant polysaccharides and immune system. Plant polysaccharides present an interesting effects as immunomodulators, particularly in the induction of the cells both in innate and adaptive immune systems. Activation of DCs could improve antitumoral responses usually diminished in cancer patients, and natural adjuvants provide a possibility of inducing this activation. ABP is a purified polysaccharide isolated from Achyranthes bidentata, a traditional Chinese medicine (TCM). The aim of this study is to investigate modulation of phenotypic and functional maturation of murine DCs by ABP. Both phenotypic and functional activities were assessed with use of conventional scanning electronic microscopy (SEM) for the morphology of the DC, transmitted electron microscopy (TEM) for intracellular lysosomes inside the DC, cellular immunohistochemistry for phagocytosis by the DCs, flow cytometry (FCM) for the changes in key surface molecules, bio-assay for the activity of acidic phosphatases (ACP), and ELISA for the production of pro-inflammatory cytokine IL-12. In fact, we found that purified ABP induced phenotypic maturation revealed by increased expression of CD86, CD40, and MHC II. Functional experiments showed the down-regulation of ACP inside DCs (which occurs when phagocytosis of DCs is decreased, and antigen presentation increased with maturation). Finally, ABP increased the production of IL-12. These data reveal that ABP promotes effective activation of murine DCs. This adjuvant-like activity may have therapeutic applications in clinical settings where immune responses need boosting. It is therefore concluded that ABP can exert positive modulation to murine DCs. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. A Cross-Sectional Study of Taenia solium in a Multiple Taeniid-Endemic Region Reveals Competition May be Protective

    Science.gov (United States)

    Conlan, James V.; Vongxay, Khamphouth; Khamlome, Boualam; Dorny, Pierre; Sripa, Banchob; Elliot, Aileen; Blacksell, Stuart D.; Fenwick, Stanley; Thompson, R. C. Andrew

    2012-01-01

    We conducted cross-sectional surveys for taeniasis and cysticercosis in humans, pigs, and dogs in four northern provinces of Laos. Human cysticercosis and taeniasis prevalence was 2.2% (95% confidence interval [CI] = 1.4–3.0%) and 8.4% (95% CI = 6.9–9.9%), respectively. Eating uncooked beef, being male, province of residence, age, and ethnicity were significant risk factors for taeniasis and only province of residence was a significant risk factor for cystiercosis. Thirty-five human tapeworms were recovered during the survey and 33 (94.3%) and 2 (5.7%) were identified as Taenia saginata and T. solium, respectively. Maximum-likelihood adjusted prevalence of T. solium and T. hydatigena in pigs was 4.2% (95% CI = 0.5–7.9%) and 55.9% (95% CI = 47.5–64.3%), respectively, and T. hydatigena taeniasis in dogs was 4.8% (95% CI = 0.0–11.3%). Taenia hydatigena and T. saginata were the most prevalent taeniids in the respective pig and human populations and together may suppress T. solium transmission. PMID:22855759

  12. Identification of cancer stem-like side population cells in purified primary cultured human laryngeal squamous cell carcinoma epithelia.

    Directory of Open Access Journals (Sweden)

    Chun-Ping Wu

    Full Text Available Cancer stem-like side population (SP cells have been identified in many solid tumors; however, most of these investigations are performed using established cancer cell lines. Cancer cells in tumor tissue containing fibroblasts and many other types of cells are much more complex than any cancer cell line. Although SP cells were identified in the laryngeal squamous cell carcinoma (LSCC cell line Hep-2 in our pilot study, it is unknown whether the LSCC tissue contains SP cells. In this study, LSCC cells (LSCCs were primary cultured and purified from a surgically resected LSCC specimen derived from a well-differentiated epiglottic neoplasm of a Chinese male. This was followed by the verification of epithelium-specific characteristics, such as ultrastructure and biomarkers. A distinct SP subpopulation (4.45±1.07% was isolated by Hoechst 33342 efflux analysis from cultured LSCCs by using a flow cytometer. Cancer stem cell (CSC-associated assays, including expression of self-renewal and CSC marker genes, proliferation, differentiation, spheroid formation, chemotherapy resistance, and tumorigenicity were then conducted between SP and non-SP (NSP LSCCs. In vitro and in vivo assays revealed that SP cells manifested preferential expression of self-renewal and CSC marker genes, higher capacity for proliferation, differentiation, and spheroid formation; enhanced resistance to chemotherapy; and greater xenograft tumorigenicity in immunodeficient mice compared with NSP cells. These findings suggest that the primary cultured and purified LSCCs contain cancer stem-like SP cells, which may serve as a valuable model for CSC research in LSCC.

  13. Incorporation of the purified epstein barr virus/C3d receptor (CR2) into liposomes and demonstration of its dual ligand binding functions

    Energy Technology Data Exchange (ETDEWEB)

    Mold, C.; Cooper, N.R.; Nemerow, G.R.

    1986-06-01

    The 145-kDA molecule that has been identified as the C3d receptor CR2 was isolated from lysates of Raji cells by affinity chromatography by using the monoclonal antibody (MoAb)HB-5. The purified protein was incorporated into /sup 14/C-phosphatidylcholine liposomes by deoxycholate dialysis followed by flotation on discontinuous sucrose gradients. Incorporation of the receptor was verified by testing the gradient fractions for CR2 by an enzyme-linked immunosorbent assay. Liposomes were shown to be unilamellar vesicles ranging in diameter from 25 to 100 nm by electron microscopy. The external orientation of CR2 in the membranes was demonstrated by immunoelectron microscopy. The functional activities of liposomes containing CR2 and liposomes without protein were compared. CR2 liposomes bound to EC3d, but not to E, and this binding was inhibited by the anti-CR2 MoAb OKB7 and by a MoAb specific for C3d. Control liposomes failed to bind to either E or EC3D. The ability of CR2 to function as a receptor for Epstein Barr virus (EBV) was tested in two ways. First, CR2 liposomes bound to B95-8, a cell line expressing EBV membrane antigens, but not to B95-8 cells treated with the viral DNA polymerase inhibitor phosphonoformic acid. Second, liposomes containing CR2 were shown by ultracentrifugal analyses to bind directly to purified EBV, and this binding was also inhibited by OKB7. Control liposomes did not bind to B95-8 cells or to EBV. These findings show that CR2 purified from detergent extracts of Raji cells can be reconstituted into lipid membranes with maintenance of its dual functions as a receptor for C3d and EBV.

  14. Incorporation of the purified epstein barr virus/C3d receptor (CR2) into liposomes and demonstration of its dual ligand binding functions

    International Nuclear Information System (INIS)

    Mold, C.; Cooper, N.R.; Nemerow, G.R.

    1986-01-01

    The 145-kDA molecule that has been identified as the C3d receptor CR2 was isolated from lysates of Raji cells by affinity chromatography by using the monoclonal antibody (MoAb)HB-5. The purified protein was incorporated into 14 C-phosphatidylcholine liposomes by deoxycholate dialysis followed by flotation on discontinuous sucrose gradients. Incorporation of the receptor was verified by testing the gradient fractions for CR2 by an enzyme-linked immunosorbent assay. Liposomes were shown to be unilamellar vesicles ranging in diameter from 25 to 100 nm by electron microscopy. The external orientation of CR2 in the membranes was demonstrated by immunoelectron microscopy. The functional activities of liposomes containing CR2 and liposomes without protein were compared. CR2 liposomes bound to EC3d, but not to E, and this binding was inhibited by the anti-CR2 MoAb OKB7 and by a MoAb specific for C3d. Control liposomes failed to bind to either E or EC3D. The ability of CR2 to function as a receptor for Epstein Barr virus (EBV) was tested in two ways. First, CR2 liposomes bound to B95-8, a cell line expressing EBV membrane antigens, but not to B95-8 cells treated with the viral DNA polymerase inhibitor phosphonoformic acid. Second, liposomes containing CR2 were shown by ultracentrifugal analyses to bind directly to purified EBV, and this binding was also inhibited by OKB7. Control liposomes did not bind to B95-8 cells or to EBV. These findings show that CR2 purified from detergent extracts of Raji cells can be reconstituted into lipid membranes with maintenance of its dual functions as a receptor for C3d and EBV

  15. Identification and characterization of a chitin-binding protein purified from coelomic fluid of the lugworm Arenicola marina defining a novel protein sequence family

    DEFF Research Database (Denmark)

    Vitashenkova, Nina; Moeller, Jesper Bonnet; Leth-Larsen, Rikke

    2012-01-01

    by coelomocytes, in the nephridium and in round cells in epidermis and in eggs. Moreover, AML-1 expression was up-regulated in response to a parasitic infection. We conclude that AML-1 purified from coelomic fluid is encoded by AML-1b and represents a novel type of protein family that binds acetylated components....

  16. Strychnos pseudoquina and Its Purified Compounds Present an Effective In Vitro Antileishmanial Activity

    Directory of Open Access Journals (Sweden)

    Paula Sousa Lage

    2013-01-01

    Full Text Available The development of new and cost-effective alternative therapeutic strategies to treat leishmaniasis has become a high priority. In the present study, the antileishmanial activity of Strychnos pseudoquina St. Hil. was investigated and pure compounds that presented this biological effect were isolated. An ethyl acetate extract was prepared, and it proved to be effective against Leishmania amazonensis. A bioactivity-guided fractionation was performed, and two flavonoids were identified, quercetin 3-O-methyl ether and strychnobiflavone, which presented an effective antileishmanial activity against L. amazonensis, and studies were extended to establish their minimum inhibitory concentrations (IC50, their leishmanicidal effects on the intra-macrophage Leishmania stage, as well as their cytotoxic effects on murine macrophages (CC50, and in O+ human red blood cells. The data presented in this study showed the potential of an ethyl acetate extract of S. pseudoquina, as well as two flavonoids purified from it, which can be used as a therapeutic alternative on its own, or in association with other drugs, to treat disease evoked by L. amazonensis.

  17. In vitro Activity and Safety Assessment of New Synthesized Thiazolo ...

    African Journals Online (AJOL)

    Purpose: To synthesis a series of novel thiazolo pyrimidine derivatives and evaluate them in vitro for their safety and anthelmintic activity against E. multilocularis metacestodes using BALB/c mice. Methods: A new series of substituted amino thiazole, hydrazinothiazole and thiazolo pyrimidine derivatives (2-6) were ...

  18. Improved radioenzymatic assay for plasma norepinephrine using purified phenylethanolamine n-methyltransferase

    International Nuclear Information System (INIS)

    Bowsher, R.R.; Henry, D.P.

    1986-01-01

    Radioenzymatic assays have been developed for catecholamines using either catechol O-methyltransferase (COMT) or phenylethanolamine N-methyltransferase (PNMT). Assays using PNMT are specific for norepinephrine (NE) and require minimal manipulative effort but until now have been less sensitive than the more complex procedures using COMT. The authors report an improved purification scheme for bovine PNMT which has permitted development of an NE assay with dramatically improved sensitivity (0.5 pg), specificity and reproducibility (C.V. < 5%). PNMT was purified by sequential pH 5.0 treatment and dialysis and by column chromatographic procedures using DEAE-Sephacel, Sepharcryl S-200 and Phenyl-Boronate Agarose. Recovery of PNMT through the purification scheme was 50%, while blank recovery was <.001%. NE can be directly quantified in 25 ul of human plasma and an 80 tube assay can be completed within 4 h. The capillary to venous plasma NE gradient was examined in 8 normotensive male subjects. Capillary plasma (NE (211.2 +/- 61.3 pg/ml)) was lower than venous plasma NE (366.6 +/- 92.5 pg/ml) in all subjects (p < 0.005). This difference suggests that capillary (NE) may be a unique indicator of sympathetic nervous system activity in vivo. In conclusion, purification of PNMT has facilitated development of an improved radioenzymatic for NE with significantly improved sensitivity

  19. Multiplex PCR identification of Taenia spp. in rodents and carnivores.

    Science.gov (United States)

    Al-Sabi, Mohammad N S; Kapel, Christian M O

    2011-11-01

    The genus Taenia includes several species of veterinary and public health importance, but diagnosis of the etiological agent in definitive and intermediate hosts often relies on labor intensive and few specific morphometric criteria, especially in immature worms and underdeveloped metacestodes. In the present study, a multiplex PCR, based on five primers targeting the 18S rDNA and ITS2 sequences, produced a species-specific banding patterns for a range of Taenia spp. Species typing by the multiplex PCR was compared to morphological identification and sequencing of cox1 and/or 12S rDNA genes. As compared to sequencing, the multiplex PCR identified 31 of 32 Taenia metacestodes from rodents, whereas only 14 cysts were specifically identified by morphology. Likewise, the multiplex PCR identified 108 of 130 adult worms, while only 57 were identified to species by morphology. The tested multiplex PCR system may potentially be used for studies of Taenia spp. transmitted between rodents and carnivores.

  20. Genetic diversity of Taenia hydatigena in the northern part of the West Bank, Palestine as determined by mitochondrial DNA sequences.

    Science.gov (United States)

    Adwan, Kamel; Jayousi, Alaa; Abuseir, Sameh; Abbasi, Ibrahim; Adwan, Ghaleb; Jarrar, Naser

    2018-06-26

    Cysticercus tenuicollis is the metacestode of canine tapeworm Taenia hydatigena, which has been reported in domestic and wild ruminants and is causing veterinary and economic losses in the meat industry. This study was conducted to determine the sequence variation in the mitochondrial cytochrome c oxidase subunit 1 (coxl) gene in 20 isolates of T. hydatigena metacestodes (cysticercus tenuicollis) collected from northern West Bank in Palestine. Nine haplotypes were detected, with one prevailing (55%). The total haplotype diversity (0.705) and the total nucleotide diversity (0.0045) displayed low genetic diversity among our isolates. Haplotype analysis showed a star-shaped network with a centrally positioned common haplotype. The Tajima's D, and Fu and Li's statistics in cysticercus tenuicollis population of this region showed a negative value, indicating deviations from neutrality and both suggested recent population expansion for the population. The findings of this study would greatly help to implement control and preventive measures for T. hydatigena larvae infection in Palestine.

  1. The effect of lignin removal by alkaline peroxide pretreatment on the susceptibility of corn stover to purified cellulolytic and xylanolytic enzymes.

    Science.gov (United States)

    Selig, Michael J; Vinzant, Todd B; Himmel, Michael E; Decker, Stephen R

    2009-05-01

    Pretreatment of corn stover with alkaline peroxide (AP) at pH 11.5 resulted in reduction of lignin content in the residual solids as a function of increasing batch temperature. Scanning electron microscopy of these materials revealed notably more textured surfaces on the plant cell walls as a result of the delignifying pretreatment. As expected, digestion of the delignified samples with commercial cellulase preparations showed an inverse relationship between the content of lignin present in the residual solids after pretreatment and the extent of both glucan and xylan conversion achievable. Digestions with purified enzymes revealed that decreased lignin content in the pretreated solids did not significantly impact the extent of glucan conversion achievable by cellulases alone. Not until purified xylanolytic activities were included with the cellulases were significant improvements in glucan conversion realized. In addition, an inverse relationship was observed between lignin content after pretreatment and the extent of xylan conversion achievable in a 24-h period with the xylanolytic enzymes in the absence of the cellulases. This observation, coupled with the direct relationship between enzymatic xylan and glucan conversion observed in a number of cases, suggests that the presence of lignins may not directly occlude cellulose present in lignocelluloses but rather impact cellulase action indirectly by its association with xylan.

  2. Generating and Purifying Fab Fragments from Human and Mouse IgG Using the Bacterial Enzymes IdeS, SpeB and Kgp.

    Science.gov (United States)

    Sjögren, Jonathan; Andersson, Linda; Mejàre, Malin; Olsson, Fredrik

    2017-01-01

    Fab fragments are valuable research tools in various areas of science including applications in imaging, binding studies, removal of Fc-mediated effector functions, mass spectrometry, infection biology, and many others. The enzymatic tools for the generation of Fab fragments have been discovered through basic research within the field of molecular bacterial pathogenesis. Today, these enzymes are widely applied as research tools and in this chapter, we describe methodologies based on bacterial enzymes to generate Fab fragments from both human and mouse IgG. For all human IgG subclasses, the IdeS enzyme from Streptococcus pyogenes has been applied to generate F(ab')2 fragments that subsequently can be reduced under mild conditions to generate a homogenous pool of Fab' fragments. The enzyme Kgp from Porphyromonas gingivalis has been applied to generate intact Fab fragments from human IgG1 and the Fab fragments can be purified using a CH1-specific affinity resin. The SpeB protease, also from S. pyogenes, is able to digest mouse IgGs and has been applied to digest antibodies and Fab fragments can be purified on light chain affinity resins. In this chapter, we describe methodologies that can be used to obtain Fab fragments from human and mouse IgG using bacterial proteases.

  3. Oocyte formation by mitotically-active germ cells purified from ovaries of reproductive age women

    Science.gov (United States)

    White, Yvonne A. R.; Woods, Dori C.; Takai, Yasushi; Ishihara, Osamu; Seki, Hiroyuki; Tilly, Jonathan L.

    2012-01-01

    Germline stem cells that produce oocytes in vitro and fertilization-competent eggs in vivo have been identified in and isolated from adult mouse ovaries. Here we describe and validate a FACS-based protocol that can be used with adult mouse ovaries and human ovarian cortical tissue to purify rare mitotically-active cells that exhibit a gene expression profile consistent with primitive germ cells. Once established in vitro, these cells can be expanded for months and spontaneously generate 35–50 µm oocytes, as determined by morphology, gene expression and attainment of haploid (1n) status. Injection of the human germline cells, engineered to stably express GFP, into human ovarian cortical biopsies leads to formation of follicles containing GFP-positive oocytes 1–2 weeks after xenotransplantation into immunodeficient female mice. Thus, ovaries of reproductive-age women, like adult mice, possess rare mitotically-active germ cells that can be propagated in vitro as well as generate oocytes in vitro and in vivo. PMID:22366948

  4. Oocyte formation by mitotically active germ cells purified from ovaries of reproductive-age women.

    Science.gov (United States)

    White, Yvonne A R; Woods, Dori C; Takai, Yasushi; Ishihara, Osamu; Seki, Hiroyuki; Tilly, Jonathan L

    2012-02-26

    Germline stem cells that produce oocytes in vitro and fertilization-competent eggs in vivo have been identified in and isolated from adult mouse ovaries. Here we describe and validate a fluorescence-activated cell sorting-based protocol that can be used with adult mouse ovaries and human ovarian cortical tissue to purify rare mitotically active cells that have a gene expression profile that is consistent with primitive germ cells. Once established in vitro, these cells can be expanded for months and can spontaneously generate 35- to 50-μm oocytes, as determined by morphology, gene expression and haploid (1n) status. Injection of the human germline cells, engineered to stably express GFP, into human ovarian cortical biopsies leads to formation of follicles containing GFP-positive oocytes 1-2 weeks after xenotransplantation into immunodeficient female mice. Thus, ovaries of reproductive-age women, similar to adult mice, possess rare mitotically active germ cells that can be propagated in vitro as well as generate oocytes in vitro and in vivo.

  5. Thermal Characterization of Purified Glucose Oxidase from A Newly Isolated Aspergillus Niger UAF-1

    Science.gov (United States)

    Anjum Zia, Muhammad; Khalil-ur-Rahman; K. Saeed, Muhammad; Andaleeb, Fozia; I. Rajoka, Muhammad; A. Sheikh, Munir; A. Khan, Iftikhar; I. Khan, Azeem

    2007-01-01

    An intracellular glucose oxidase was isolated from the mycelium extract of a locally isolated strain of Aspergillus niger UAF-1. The enzyme was purified to a yield of 28.43% and specific activity of 135 U mg−1 through ammonium sulfate precipitation, anion exchange and gel filtration chromatography. The enzyme showed high affinity for D-glucose with a Km value of 2.56 mM. The enzyme exhibited optimum catalytic activity at pH 5.5. Temperature optimum for glucose oxidase, catalyzed D-glucose oxidation was 40°C. The enzyme showed a high thermostability having a half-life 30 min, enthalpy of denaturation 99.66 kJ mol−1 and free energy of denaturation 103.63 kJ mol−1. These characteristics suggest the use of glucose oxidase from Aspergillus niger UAF-1 as an analytical reagent and in the design of biosensors for clinical, biochemical and diagnostic assays. PMID:18193107

  6. Comparative analysis of complete mitochondrial genomes suggests that relaxed purifying selection is driving high nonsynonymous evolutionary rate of the NADH2 gene in whitefish (Coregonus ssp.)

    DEFF Research Database (Denmark)

    Jacobsen, Magnus W.; Rodrigues da Fonseca, Rute Andreia; Bernatchez, Louis

    2016-01-01

    (Coregonus ssp.) for signals of positive selection. These salmonids show several distinct morphological and ecological differences that may be associated with energetics and therefore potentially positive selection at the mitogenome level. We found that purifying selection and genetic drift were...

  7. Antisense myb inhibition of purified erythroid progenitors in development and differentiation is linked to cycling activity and expression of DNA polymerase alpha

    International Nuclear Information System (INIS)

    Valtieri, M.; Venturelli, D.; Care, A.; Fossati, C.; Pelosi, E.; Labbaye, C.; Mattia, G.; Gewirtz, A.M.; Calabretta, B.; Peschle, C.

    1991-01-01

    These studies aimed to determine the expression and functional role of c-myb in erythroid progenitors with different cycling activities. In the first series of experiments the erythroid burst-forming unit (BFU-E) and colony-forming unit (CFU-E) populations from adult peripheral blood (PB), bone marrow (BM), and embryonic-fetal liver (FL) were treated with either c-myb antisense oligomers or 3H-thymidine (3H-TdR). A direct correlation was always observed between the inhibitory effect of anti-myb oligomers and the level of cycling activity. Thus, the inhibitory effect of antisense c-myb on the number of BFU-E colonies was 28.3% +/- 15.8% in PB, 53.4% +/- 9.3% in BM, and 68.2% +/- 24.5% in FL. Both adult and embryonic CFU-E were markedly inhibited. Using purified PB progenitors, we observed a similar pattern, although with slightly lower inhibitory effects. In the 3H-TdR suicide assay the killing index of BFU-E was 8.9% +/- 4.2% in PB, 29.4% +/- 6.5% in BM, and 40.1% +/- 9.6% in FL. The values for adult and embryonic CFU-E were 55.7% +/- 7.9% and 60.98% +/- 6.6%, respectively. We then investigated the kinetics of c-myb mRNA level during the erythroid differentiation of purified adult PB and FL BFU-E, as evaluated in liquid-phase culture by reverse transcription-polymerase chain reaction. Adult erythroid precursors showed a gradual increase of c-myb mRNA from day 4 through day 8 of culture and a sharp decrease at later times, whereas the expression of c-myb mRNA and protein in differentiation embryonic precursors peaked 2 days earlier. In both cases, c-myb mRNA level peaked at the CFU-E stage of differentiation. Finally, highly purified adult PB BFU-E were stimulated into cycling by a 3-day treatment with interleukin-3 in liquid phase: both the sensitivity to c-myb antisense oligomers and the 3H-TdR suicide index showed a gradual, strictly parallel increase

  8. The interaction between beta 2-microglobulin (beta 2m) and purified class-I major histocompatibility (MHC) antigen

    DEFF Research Database (Denmark)

    Pedersen, L O; Hansen, A S; Olsen, A C

    1994-01-01

    been generated recently and this paper reports on a similar assay for the interaction between beta 2m and class I. As a model system human beta 2m binding to mouse class I was used. The assay is strictly biochemical using purified reagents which interact in solution and complex formation is determined...... by size separation. It is specific and highly sensitive. The observed affinity of the interaction, KD, is close to 0.4 nM. The rate of association at 37 degrees C is very fast (the ka is around 5 x 10(4)/M/s) whereas the dissociation is slow (the kd is around 8 x 10(-6)/s); the ratio of dissociation...

  9. Effect of honey on oxidation, chlorination and nitration by purified equine myeloperoxidase

    Directory of Open Access Journals (Sweden)

    Saad Aissat

    2017-09-01

    Full Text Available Objective: To evaluate the antioxidant effect of honey using two classical methods generally used, and for the first time to test the effect of honey on the oxidation, chlorination and nitration by purified equine myeloperoxidase (MPO. Methods: The antioxidant activity of three Algerian honey samples (nectar honey, mixed honey and honeydew honey was evaluated by two classical methods, the ferric- reducing/antioxidant power (FRAP assay and 2,2-diphenyl-1-picrylhydrazyl (DPPH radical-scavenging capacity. Results: Honeydew honey had the highest reducing power and DPPH radical-scavenging activity, whereas nectar honey showed the lowest reducing power and DPPH radical-scavenging activity. All honey samples showed a significant inhibitory effect on the chlorination activity of equine MPO, but honeydew honey was the weakest inhibitor. The three samples were poorly inhibitor on the MPO oxidation and nitration activities, except for nectar honey that exerted an inhibitory effect at the highest tested concentration of 10%. These later results seem to contradict those obtained with DPPH and FRAP. Conclusions: The antioxidant capacity of honey is mainly due to the phenolic compounds and flavonoids it contained. It has been suggested that MPO might be involved in the antioxidant, not pro-oxidant, activity of phenolic compounds.

  10. Mycobacterium avium and purified protein derivative-specific cytotoxicity mediated by CD4+ lymphocytes from healthy HIV-seropositive and-seronegative individuals

    DEFF Research Database (Denmark)

    Ravn, P; Pedersen, B K

    1996-01-01

    mycobacteria. Our objective was to investigate the M.tuberculosis-and M. avium-specific cytotoxic capacity of T cells from healthy, bacille Calmette-Guérin-vaccinated, HIV-seropositive individuals. Blood mononuclear cells were obtained from 10 healthy HIV-seropositive and 10 healthy seronegative persons...... with no history of previous or active mycobacterial infection. Antigen-specific killing of macrophages presenting mycobacterial antigens (purified protein derivative or M. avium culture filtrate) was conducted. The phenotype of the killer cells was determined by a fluorescence-activated cell sorter after antigen...

  11. Measles virus polypeptides in purified virions and in infected cells

    International Nuclear Information System (INIS)

    Vainionpaeae, R.; Ziola, B.; Salmi, A.

    1978-01-01

    A wild-type measles virus was radiolabeled during growth in VERO cells and purified by two successive potassium tartrate gradient centrifugations. The virion polypeptide composition was determined by SDS-polyacrylamide gel electrophoresis employing two different buffer systems. Six virus-specific polypeptides were consistently detected. The largest (L) had a molecular weight (MW) of greater than 150,000. The second largest polypeptide, G (MW 79,000), was the only glycoprotein found. The proteins designated polypeptide 2 (MW 66 to 70,000) and nucleocapsid protein or NP (MW 61,000) were phosphorylated. The remaining virus-coded proteins were polypeptide 5 (MW 40,000) and the matrix or M protein (MW 37,000). Measles virions also contained a polypeptide (MW 42,000) thought to be actin due to co-migration with this component of uninfected cells. Analysis of in vitro 3 H-acetic anhydride radiolabeled virions confirmed the presence of these seven polypeptides. Acetic anhydride also labeled a protein designated polypeptide 4 (MW 53,000) which was not consistently radiolabeled in vivo, as well as several other minor proteins believed to be cellular in origin. Synthesis of the six virus-specific structural polypeptides was detected in lysates of infected cells by SDS-polyacrylamide slab gel electrophoresis. Virus specificity of polypeptide 4 could not be confirmed due to the similar MW of several cellular polypeptides. Two non-virion, but virus-specified polypeptides, of MW 38,000 and 18,000 were also detected. Synthesis of the virus structural proteins was in the same proportions as the polypeptides found in virions except for under production of polypeptide G and over production of polypeptide 2. (author)

  12. DNA/RNA hybrid substrates modulate the catalytic activity of purified AID.

    Science.gov (United States)

    Abdouni, Hala S; King, Justin J; Ghorbani, Atefeh; Fifield, Heather; Berghuis, Lesley; Larijani, Mani

    2018-01-01

    Activation-induced cytidine deaminase (AID) converts cytidine to uridine at Immunoglobulin (Ig) loci, initiating somatic hypermutation and class switching of antibodies. In vitro, AID acts on single stranded DNA (ssDNA), but neither double-stranded DNA (dsDNA) oligonucleotides nor RNA, and it is believed that transcription is the in vivo generator of ssDNA targeted by AID. It is also known that the Ig loci, particularly the switch (S) regions targeted by AID are rich in transcription-generated DNA/RNA hybrids. Here, we examined the binding and catalytic behavior of purified AID on DNA/RNA hybrid substrates bearing either random sequences or GC-rich sequences simulating Ig S regions. If substrates were made up of a random sequence, AID preferred substrates composed entirely of DNA over DNA/RNA hybrids. In contrast, if substrates were composed of S region sequences, AID preferred to mutate DNA/RNA hybrids over substrates composed entirely of DNA. Accordingly, AID exhibited a significantly higher affinity for binding DNA/RNA hybrid substrates composed specifically of S region sequences, than any other substrates composed of DNA. Thus, in the absence of any other cellular processes or factors, AID itself favors binding and mutating DNA/RNA hybrids composed of S region sequences. AID:DNA/RNA complex formation and supporting mutational analyses suggest that recognition of DNA/RNA hybrids is an inherent structural property of AID. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Bitistatin-functionalized fluorescent nanodiamond particles specifically bind to purified human platelet integrin receptor αIIbβ3 and activated platelets

    Directory of Open Access Journals (Sweden)

    Marcinkiewicz C

    2017-05-01

    Full Text Available Cezary Marcinkiewicz,1,2 Jonathan A Gerstenhaber,1 Mark Sternberg,2 Peter I Lelkes,1 Giora Feuerstein1,2 1Department of Bioengineering, College of Engineering, Temple University, Philadelphia, 2Debina Diagnostic, Inc., Newton Square, PA, USA Abstract: Thromboembolic events (TEE underwrite key causes of death in developed countries. While advanced imaging technologies such as computed tomography scans serve to diagnose blood clots during acute cardiovascular events, no such technology is available in routine primary care for TEE risk assessment. Here, we describe an imaging platform technology based on bioengineered fluorescent nanodiamond particles (F-NDPs functionalized with bitistatin (Bit, a disintegrin that specifically binds to the αIIbβ3 integrin, platelet fibrinogen receptor (PFR on activated platelets. Covalent linkage of purified Bit to F-NDP was concentration-dependent and saturable, as validated by enzyme-linked immunosorbent assay using specific anti-Bit antibodies. F-NDP–Bit interacted with purified PFR, either in immobilized or soluble form. Lotrafiban, a nonpeptide, αIIbβ3 receptor antagonist, specifically blocked F-NDP–Bit–PFR complex formation. Moreover, F-NDP–Bit specifically binds to activated platelets incorporated into a clot generated by thrombin-activated rat platelet-rich plasma (PRP. Our results suggest that engineered F-NDP–Bit particles could serve as noninvasive, “real-time” optical diagnostics for clots present in blood vessels. Keywords: carbon nanoparticles, blood clots, imaging, platelet fibrinogen receptor, fluorescence, disintegrin, thromboembolic complications, thrombosis

  14. Modulation in the activity of purified tonoplast H+-ATPase by tonoplast glycolipids prepared from cultured rice (Oryza sativa L. var. Boro) cells.

    Science.gov (United States)

    Yamaguchi, M; Kasamo, K

    2001-05-01

    Glycolipids, phospholipids, and neutral lipids were extracted from the tonoplast fraction of cultured rice cells (Oryza sativa L. var. Boro). Acyl steryl glucoside (ASG) and glucocerebroside (GlcCer) were also prepared from this fraction. We determined the effects of these tonoplast lipids on the activity of H+-ATPase which was delipidated and purified from the tonoplast fraction. Exogenously added tonoplast phospholipids stimulated the activity of purified tonoplast H+-ATPase, but tonoplast glycolipids did not. When tonoplast glycolipids or tonoplast ASG was added in the presence of tonoplast phospholipids, they decreased the phospholipid-induced activation of the tonoplast H+-ATPase; tonoplast GlcCer only caused a small decrease. Steryl glucoside (SG) did not cause any decrease in this activation. Phospholipids, ASG, and GlcCer made up 35 mol%, 20 mol% and 7 mol% of the total lipids of the tonoplast fraction of cultured rice cells, respectively, and these glycolipid levels were enough to depress the phospholipid-induced activation of the tonoplast H+-ATPASE: These results revealed that H+-ATPase activity in the tonoplast may be modulated toward activation and depression by tonoplast phospholipids and glycolipids, respectively. The acylation of SG would be responsible for the depression in the phospholipid-induced H+-ATPase activity.

  15. Indium-111 labeled purified granulocytes in the diagnosis of synthetic vascular graft infection

    International Nuclear Information System (INIS)

    Forstrom, L.A.; Dewanjee, M.K.; Chowdhury, S.; Brown, M.L.

    1988-01-01

    Indium-111 labeled leukocytes have been shown to be useful in the diagnosis of synthetic vascular graft infection. To minimize the potential effects of labeled red blood cells and platelets on image interpretation, the authors prepared purified autologous granulocytes (PG) from 84 ml of blood using Volex enhanced gravity sedimentation and Ficoll-Hypaque double density centrifugation. The labeling efficiency of PG with In-111 tropolone was 90 +/- 9% (mean +/- SD). Imaging was performed 18-24 hours following injection of approximately 445 microcuries of In-111 PG in 26 patients with suspected infection of vascular grafts that had been implanted 12 days to 12 years prior to the study. In ten patients with proven graft infection, seven had positive In-111 PG scans. Ten of 11 patients without infection had negative scans. In five patients with clinically equivocal findings, scan results were positive in one, negative in one, and equivocal in three. A false-positive scan occurred in a patient with an uninfected inflammatory pseudoaneurysm of an aortic graft. These results confirm an earlier report that In-111 PG imaging is a useful technique in the diagnosis of synthetic vascular graft infection

  16. HARP preferentially co-purifies with RPA bound to DNA-PK and blocks RPA phosphorylation.

    Science.gov (United States)

    Quan, Jinhua; Yusufzai, Timur

    2014-05-01

    The HepA-related protein (HARP/SMARCAL1) is an ATP-dependent annealing helicase that is capable of rewinding DNA structures that are stably unwound due to binding of the single-stranded DNA (ssDNA)-binding protein Replication Protein A (RPA). HARP has been implicated in maintaining genome integrity through its role in DNA replication and repair, two processes that generate RPA-coated ssDNA. In addition, mutations in HARP cause a rare disease known as Schimke immuno-osseous dysplasia. In this study, we purified HARP containing complexes with the goal of identifying the predominant factors that stably associate with HARP. We found that HARP preferentially interacts with RPA molecules that are bound to the DNA-dependent protein kinase (DNA-PK). We also found that RPA is phosphorylated by DNA-PK in vitro, while the RPA-HARP complexes are not. Our results suggest that, in addition to its annealing helicase activity, which eliminates the natural binding substrate for RPA, HARP blocks the phosphorylation of RPA by DNA-PK.

  17. AKUT-II: an experimental plant for purifying the HTR loop of combustion waste gas

    Energy Technology Data Exchange (ETDEWEB)

    Beaujean, H.; Vygen, H.

    1976-02-15

    A plant for the separation of aerosols, krypton and tritium (AKUT) used for purifying the head end of the reprocessing of thorium-containing fuel elements from combustion waste gases is described. Data are to be collected to enable a process engineer to plan and construct a large-scale plant, and the correctness and practicability of the concept adopted is to be proved in conjunction with the JUPITER plant. It is true that the tests on the AKUT I plant confirmed that the flow scheme was basically correct, but the actual experimental operation was considerably limited by a fixed and rigid coupling to the combustion furnace. Some operational conditions were encountered which did not meet the design values. Part of the plant (krypton separation) is being tested in the USA. The German concept was taken over in the early stages of tests and adapted to existing apparatuses, the result inevitably being different experimental conditions. The AKUT II plant can now be used for consideration of the economic and safety conditions, and comparisons can be made.

  18. Cardiopulmonary benefits of reducing indoor particles of outdoor origin: a randomized, double-blind crossover trial of air purifiers.

    Science.gov (United States)

    Chen, Renjie; Zhao, Ang; Chen, Honglei; Zhao, Zhuohui; Cai, Jing; Wang, Cuicui; Yang, Changyuan; Li, Huichu; Xu, Xiaohui; Ha, Sandie; Li, Tiantian; Kan, Haidong

    2015-06-02

    Indoor exposure to fine particulate matter (PM2.5) from outdoor sources is a major health concern, especially in highly polluted developing countries such as China. Few studies have evaluated the effectiveness of indoor air purification on the improvement of cardiopulmonary health in these areas. This study sought to evaluate whether a short-term indoor air purifier intervention improves cardiopulmonary health. We conducted a randomized, double-blind crossover trial among 35 healthy college students in Shanghai, China, in 2014. These students lived in dormitories that were randomized into 2 groups and alternated the use of true or sham air purifiers for 48 h with a 2-week washout interval. We measured 14 circulating biomarkers of inflammation, coagulation, and vasoconstriction; lung function; blood pressure (BP); and fractional exhaled nitric. We applied linear mixed-effect models to evaluate the effect of the intervention on health outcome variables. On average, air purification resulted in a 57% reduction in PM2.5 concentration, from 96.2 to 41.3 μg/m3, within hours of operation. Air purification was significantly associated with decreases in geometric means of several circulating inflammatory and thrombogenic biomarkers, including 17.5% in monocyte chemoattractant protein-1, 68.1% in interleukin-1β, 32.8% in myeloperoxidase, and 64.9% in soluble CD40 ligand. Furthermore, systolic BP, diastolic BP, and fractional exhaled nitrous oxide were significantly decreased by 2.7%, 4.8%, and 17.0% in geometric mean, respectively. The impacts on lung function and vasoconstriction biomarkers were beneficial but not statistically significant. This intervention study demonstrated clear cardiopulmonary benefits of indoor air purification among young, healthy adults in a Chinese city with severe ambient particulate air pollution. (Intervention Study on the Health Impact of Air Filters in Chinese Adults; NCT02239744). Copyright © 2015 American College of Cardiology Foundation

  19. Optimization of freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan) peel.

    Science.gov (United States)

    Mehrnoush, Amid; Mustafa, Shuhaimi; Yazid, Abdul Manap Mohd

    2012-01-01

    Response surface methodology (RSM) along with central composite design (CCD) was applied to optimize the freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan) peel. The effect of pectinase content (-2.66, 62.66 mg/mL), Arabic gum (-1.21, 10.21%, w/v), and maltodextrin (0.73, 7.26%, w/v) as independent variables on activity, yield, and storage stability of freeze-dried enzyme was evaluated. Storage stability of pectinase was investigated after one week at 4 °C and yield percentage of the enzyme after encapsulation was also determined. The independent variables had the most significant (p mango peel were obtained using 30 mg/mL of pectinase content, 4.5 (%, w/v) of Arabic gum, and 4 (%, w/v) of maltodextrin. Under these conditions, the maximum activity (11.12 U/mL), yield (86.4%) and storage stability (84.2%) of encapsulated pectinase were achieved.

  20. Cellular cytotoxic response induced by highly purified multi-wall carbon nanotube in human lung cells.

    Science.gov (United States)

    Tsukahara, Tamotsu; Haniu, Hisao

    2011-06-01

    Carbon nanotubes, a promising nanomaterial with unique characteristics, have applications in a variety of fields. The cytotoxic effects of carbon nanotubes are partially due to the induction of oxidative stress; however, the detailed mechanisms of nanotube cytotoxicity and their interaction with cells remain unclear. In this study, the authors focus on the acute toxicity of vapor-grown carbon fiber, HTT2800, which is one of the most highly purified multi-wall carbon nanotubes (MWCNT) by high-temperature thermal treatment. The authors exposed human bronchial epithelial cells (BEAS-2B) to HTT2800 and measured the cellular uptake, mitochondrial function, cellular LDH release, apoptotic signaling, reactive oxygen species (ROS) generation and pro-inflammatory cytokine release. The HTT2800-exposed cells showed cellular uptake of the carbon nanotube, increased cell death, enhanced DNA damage, and induced cytokine release. However, the exposed cells showed no obvious intracellular ROS generation. These cellular and molecular findings suggest that HTT2800 could cause a potentially adverse inflammatory response in BEAS-2B cells.

  1. Steviol glycosides in purified stevia leaf extract sharing the same metabolic fate.

    Science.gov (United States)

    Purkayastha, Sidd; Markosyan, Avetik; Prakash, Indra; Bhusari, Sachin; Pugh, George; Lynch, Barry; Roberts, Ashley

    2016-06-01

    The safety of steviol glycosides is based on data available on several individual steviol glycosides and on the terminal absorbed metabolite, steviol. Many more steviol glycosides have been identified, but are not yet included in regulatory assessments. Demonstration that these glycosides share the same metabolic fate would indicate applicability of the same regulatory paradigm. In vitro incubation assays with pooled human fecal homogenates, using rebaudiosides A, B, C, D, E, F and M, as well as steviolbioside and dulcoside A, at two concentrations over 24-48 h, were conducted to assess the metabolic fate of various steviol glycoside classes and to demonstrate that likely all steviol glycosides are metabolized to steviol. The data show that glycosidic side chains containing glucose, rhamnose, xylose, fructose and deoxy-glucose, including combinations of α(1-2), β-1, β(1-2), β(1-3), and β(1-6) linkages, were degraded to steviol mostly within 24 h. Given a common metabolite structure and a shared metabolic fate, safety data available for individual steviol glycosides can be used to support safety of purified steviol glycosides in general. Therefore, steviol glycosides specifications adopted by the regulatory authorities should include all steviol glycosides belonging to the five groups of steviol glycosides and a group acceptable daily intake established. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Simple Identification of Human Taenia Species by Multiplex Loop-Mediated Isothermal Amplification in Combination with Dot Enzyme-Linked Immunosorbent Assay

    Science.gov (United States)

    Nkouawa, Agathe; Sako, Yasuhito; Okamoto, Munehiro; Ito, Akira

    2016-01-01

    For differential detection of Taenia solium, Taenia saginata, and Taenia asiatica, loop-mediated isothermal amplification (LAMP) assay targeting the cytochrome c oxidase subunit 1 gene has been recently developed and shown to be sensitive, specific, and effective. However, to achieve differential identification, one specimen requires three reaction mixtures containing a primer set of each Taenia species separately, which is complex and time consuming and increases the risk of cross-contamination. In this study, we developed a simple differential identification of human Taenia species using multiplex LAMP (mLAMP) in combination with dot enzyme-linked immunosorbent assay (dot-ELISA). Forward inner primers of T. solium, T. saginata, and T. asiatica labeled with fluorescein isothiocyanate (FITC), digoxigenin (DIG), and tetramethylrhodamine (TAMRA), respectively, and biotin-labeled backward inner primers were used in mLAMP. The mLAMP assay succeeded in specific amplification of each respective target gene in a single tube. Furthermore, the mLAMP product from each species was easily distinguished by dot-ELISA with an antibody specific for FITC, DIG, or TAMRA. The mLAMP assay in combination with dot-ELISA will make identification of human Taenia species simpler, easier, and more practical. PMID:27044566

  3. Use of anionic denaturing detergents to purify insoluble proteins after overexpression

    Directory of Open Access Journals (Sweden)

    Schlager Benjamin

    2012-12-01

    Full Text Available Background Many proteins form insoluble protein aggregates, called “inclusion bodies”, when overexpressed in E. coli. This is the biggest obstacle in biotechnology. Ever since the reversible denaturation of proteins by chaotropic agents such as urea or guanidinium hydrochloride had been shown, these compounds were predominantly used to dissolve inclusion bodies. Other denaturants exist but have received much less attention in protein purification. While the anionic, denaturing detergent sodiumdodecylsulphate (SDS is used extensively in analytical SDS-PAGE, it has rarely been used in preparative purification. Results Here we present a simple and versatile method to purify insoluble, hexahistidine-tagged proteins under denaturing conditions. It is based on dissolution of overexpressing bacterial cells in a buffer containing sodiumdodecylsulfate (SDS and whole-lysate denaturation of proteins. The excess of detergent is removed by cooling and centrifugation prior to affinity purification. Host- and overexpressed proteins do not co-precipitate with SDS and the residual concentration of detergent is compatible with affinity purification on Ni/NTA resin. We show that SDS can be replaced with another ionic detergent, Sarkosyl, during purification. Key advantages over denaturing purification in urea or guanidinium are speed, ease of use, low cost of denaturant and the compatibility of buffers with automated FPLC. Conclusion Ionic, denaturing detergents are useful in breaking the solubility barrier, a major obstacle in biotechnology. The method we present yields detergent-denatured protein. Methods to refold proteins from a detergent denatured state are known and therefore we propose that the procedure presented herein will be of general application in biotechnology.

  4. Steroid metabolism by purified adult rat Leydig cells in primary culture

    International Nuclear Information System (INIS)

    Browning, J.Y.; Tcholakian, R.K.; Kessler, M.J.; Grotjan, H.E. Jr.

    1982-01-01

    To characterize Leydig cell steroidogensis, we examined the metabolism of [3H]pregnenolone (3 beta-hydroxy-5-pregnen-20-one) to androgens in the presence and absence of human chorionic gonadotropin (hCG) as a function of culture duration. Approximately 20-30% of the (3H)pregnenolone was converted to testosterone (17 beta-hydroxy-4-androsten-3-one) by purified Leydig cells at 0, 3 and 5 days (d) of culture. Androstenedione (4-androstene-3,17-dione) and dihydrotestosterone (17 beta-hydroxy-5 alpha-androstan-3-one) were also produced while on day 5 of culture, significant amounts of progesterone (4-pregnene-3,20-dione) were isolated. The delta 5 intermediates, 17-hydroxypregnenolone (3 beta, 17-dihydroxy-5-pregnen-20-one) and dehydroepiandrosterone (3 beta-hydroxy-5-androsten-17-one), accounted for less than 1% of substrate conversion, indicating a clear preference for Leydig cells to metabolize (3H)pregnenolone via the delta 4 pathway. On day 0 of culture, unidentified metabolites considered of predominately polar steroids while on day 5 of culture, the unidentified metabolites consisted of predominately nonpolar steroids. In the presence of hCG, (3H-pregnenolone metabolism did not differ from basal on day 0 or 3 of culture. HCG increased the conversion of pregnenolone to progesterone and 17-hydroxyprogesterone (17-hydroxy-4-pregnene-3,20-dione) on 5d. This suggests that Leydig cells cultured for 5d have decreased C17-20 desmolase activity or that hCG acutely stimulates 3 beta-hydroxysteroid dehydrogenase and delta 5-delta 5 isomerase activities

  5. The industrial applicability of purified cellulase complex indigenously produced by Trichoderma viride through solid-state bio-processing of agro-industrial and municipal paper wastes

    Directory of Open Access Journals (Sweden)

    Muhammad Irshad

    2013-02-01

    Full Text Available An indigenous strain of Trichoderma viride produced high titers of cellulase complex in solid-state bio-processing of agro-industrial orange peel waste, which was used as the growth-supporting substrate. When the conditions of the SSF medium containing 15 g orange peel (50% w/w moisture inoculated with 5 mL of inoculum were optimal, the maximum productions of endoglucanase (655 ± 5.5 U/mL, exoglucanase (412 ± 4.3 U/mL, and β-glucosidase (515 ± 3.7 U/mL were recorded after 4 days of incubation at pH 5 and 35 °C. The enzyme with maximum activity (endoglucanase was purified by ammonium sulfate fractionation and Sephadex G-100 column gel filtration chromatographic technique. Endoglucanase was 5.5-fold purified with specific activity of 498 U/mg in comparison to the crude enzyme. The enzyme was shown to have a molecular weight of 58 kDa by sodium dodecyl sulphate poly-acrylamide gel electrophoresis (SDS-PAGE. The shelf life profile revealed that the enzyme could be stored at room temperature (30 °C for up to 45 days without losing much of its activity.

  6. Identification of UDPG-binding polypeptides and purified (1,3)-β-glucan synthase by photoaffinity labelling with 5-azido-UDPG

    International Nuclear Information System (INIS)

    Frost, D.J.; Wu, A.; Read, S.M.; Wasserman, B.P.; Drake, R.R.; Haley, B.E.

    1989-01-01

    The photoaffinity probe 5-azido-uridine 5'-β-[ 32 P]-diphosphate glucose was used to identify the major UDPG-binding polypeptide of red beet (1,3)-β-glucan synthase. Glucan synthase was purified from plasma membranes by sequential solubilization with CHAPS followed by product entrapment. Two major polypeptides at 72 and 54 kD were labelled by probe. Labelling of both was abolished with increasing levels of cold UDPG. However, labelling of the 54 kD polypeptide was dependent upon the presence of divalent cations. These data suggest that the 54 kD polypeptide is a substrate-binding and cation-regulated component of the glucan synthase complex

  7. Purifying capability, enzyme activity, and nitrification potentials in December in integrated vertical flow constructed wetland with earthworms and different substrates.

    Science.gov (United States)

    Xu, Defu; Gu, Jiaru; Li, Yingxue; Zhang, Yu; Howard, Alan; Guan, Yidong; Li, Jiuhai; Xu, Hui

    2016-01-01

    The response of purifying capability, enzyme activity, nitrification potentials, and total number of bacteria in the rhizosphere in December to wetland plants, substrates, and earthworms was investigated in integrated vertical flow constructed wetlands (IVFCW). The removal efficiency of total nitrogen (TN), NH4-N, chemical oxygen demand (COD), and total phosphorus (TP) was increased when earthworms were added into IVFCW. A significantly average removal efficiency of N in IVFCW that employed river sand as substrate and in IVFCW that employed a mixture of river sand and Qing sand as substrate was not found. However, the average removal efficiency of P was higher in IVFCW with a mixture of river sand and Qing sand as substrate than in IVFCW with river sand as substrate. Invertase activity in December was higher in IVFCW that used a mixture of river sand and Qing sand as substrate than in IVFCW which used only river sand as substrate. However, urease activity, nitrification potential, and total number of bacteria in December was higher in IVFCW that employed river sand as substrate than in IVFCW with a mixture of river sand and Qing sand as substrate. The addition of earthworms into the integrated vertical flow constructed wetland increased the above-ground biomass, enzyme activity (catalase, urease, and invertase), nitrification potentials, and total number of bacteria in December. The above-ground biomass of wetland plants was significantly positively correlated with urease and nitrification potentials (p earthworms into IVFCW increased enzyme activity and nitrification potentials in December, which resulted in improving purifying capability.

  8. Targeting Echinococcus multilocularis stem cells by inhibition of the Polo-like kinase EmPlk1.

    Directory of Open Access Journals (Sweden)

    Andreas Schubert

    2014-06-01

    Full Text Available Alveolar echinococcosis (AE is a life-threatening disease caused by larvae of the fox-tapeworm Echinococcus multilocularis. Crucial to AE pathology is continuous infiltrative growth of the parasite's metacestode stage, which is driven by a population of somatic stem cells, called germinative cells. Current anti-AE chemotherapy using benzimidazoles is ineffective in eliminating the germinative cell population, thus leading to remission of parasite growth upon therapy discontinuation.We herein describe the characterization of EmPlk1, encoded by the gene emplk1, which displays significant homologies to members of the Plk1 sub-family of Polo-like kinases that regulate mitosis in eukaryotic cells. We demonstrate germinative cell-specific expression of emplk1 by RT-PCR, transcriptomics, and in situ hybridization. We also show that EmPlk1 can induce germinal vesicle breakdown when heterologously expressed in Xenopus oocytes, indicating that it is an active kinase. This activity was significantly suppressed in presence of BI 2536, a Plk1 inhibitor that has been tested in clinical trials against cancer. Addition of BI 2536 at concentrations as low as 20 nM significantly blocked the formation of metacestode vesicles from cultivated Echinococcus germinative cells. Furthermore, low concentrations of BI 2536 eliminated the germinative cell population from mature metacestode vesicles in vitro, yielding parasite tissue that was no longer capable of proliferation.We conclude that BI 2536 effectively inactivates E. multilocularis germinative cells in parasite larvae in vitro by direct inhibition of EmPlk1, thus inducing mitotic arrest and germinative cell killing. Since germinative cells are decisive for parasite proliferation and metastasis formation within the host, BI 2536 and related compounds are very promising compounds to complement benzimidazoles in AE chemotherapy.

  9. Production of Antimicrobial Films by Incorporation of Partially Purified Lysozyme into Biodegradable Films of Crude Exopolysaccharides Obtained from Aureobasidium pullulans Fermentation

    Directory of Open Access Journals (Sweden)

    Nilay Kandemir

    2005-01-01

    Full Text Available Antimicrobial films were produced by incorporating partially purified lysozyme into films of crude exopolysaccharides (59 % pullulan obtained from Aureobasidium pullulans fermentation. After film making, the films containing lysozyme at 100, 260, 520 and 780 μg/cm2 showed 23 to 70 % of their expected enzyme activities. The highest recovery of enzyme activity (65–70 % after the film making was obtained in films prepared by incorporating lysozyme at 260 μg/cm2 (1409 U/cm2. The incorporation of disodium EDTA×2H2O and sucrose did not affect the initial lysozyme activity of the films significantly. With or without the presence of disodium EDTA×2H2O at 52 or 520 μg/cm2, lysozyme activity showed sufficient stability in the films during 21 days of cold storage. However, the presence of sucrose at 10 mg/cm2 in the films caused the destabilization of part of enzyme activity (almost 35 % at the end of storage. The combinational incorporation of lysozyme at 780 μg/cm2 (4227 U/cm2 and disodium EDTA×2H2O at 520 μg/cm2 gave antimicrobial films effective on Escherichia coli. However, in the studied lysozyme concentration range the films did not show any antimicrobial activity against Lactobacillus plantarum. This study clearly showed that the partially purified lysozyme and crude exopolysaccharides from Aureobasidium pullulans may be used to obtain antimicrobial films to increase the safety of foods.

  10. Larval gryporhynchid tapeworms (Cestoda: Cyclophyllidea) of British freshwater fish, with a description of the pathology caused by Paradilepis scolecina

    Czech Academy of Sciences Publication Activity Database

    Williams, C. F.; Reading, A. J.; Scholz, Tomáš; Shinn, A. P.

    2012-01-01

    Roč. 86, č. 1 (2012), 1-9 ISSN 0022-149X R&D Projects: GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : CARP CYPRINUS-CARPIO * METACESTODES * DILEPIDIDAE * PARASITES * PLEROCERCOIDS * MOZAMBIQUE * INFECTION * LINNAEUS * LAKE Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.157, year: 2012

  11. Identification of proteins from tuberculin purified protein derivative (PPD) by LC-MS/MS.

    Science.gov (United States)

    Borsuk, Sibele; Newcombe, Jane; Mendum, Tom A; Dellagostin, Odir A; McFadden, Johnjoe

    2009-11-01

    The tuberculin purified protein derivative (PPD) is a widely used diagnostic antigen for tuberculosis, however it is poorly defined. Most mycobacterial proteins are extensively denatured by the procedure employed in its preparation, which explains previous difficulties in identifying constituents from PPD to characterize their behaviour in B- and T-cell reactions. We here described a proteomics-based characterization of PPD from several different sources by LC-MS/MS, which combines the solute separation power of HPLC, with the detection power of a mass spectrometer. The technique is able to identify proteins from complex mixtures of peptide fragments. A total of 171 different proteins were identified among the four PPD samples (two bovine PPD and two avium PPD) from Brazil and UK. The majority of the proteins were cytoplasmic (77.9%) and involved in intermediary metabolism and respiration (24.25%) but there was a preponderance of proteins involved in lipid metabolism. We identified a group of 21 proteins that are present in both bovine PPD but were not detected in avium PPD preparation. In addition, four proteins found in bovine PPD are absent in Mycobacterium bovis BCG vaccine strain. This study provides a better understanding of the tuberculin PPD components leading to the identification of additional antigens useful as reagents for specific diagnosis of tuberculosis.

  12. Kajian Aspek Epidemiologi Taeniasis dan Sistiserkosis di Papua

    OpenAIRE

    Sandi, Semuel

    2014-01-01

    . Taeniasis merupakan infeksi pada saluran pencernaan oleh cacing dewasaTaenia solium, Taenia saginata dan Taenia asiaticasedangkan sistiserkosis merupakan penyakit/infeksi pada jaringan lunak yang disebabkan oleh larva Taenia solium. Penyakit ini masih ditemukan di Indonesia khusunya Provinsi Bali, Sumatra Utara dan Papua dengan kisaran prevalensi 2–48%. Analisis data epidemiologi penyakit taeniasis dan sistiserkosis sangat diperlukan untuk memahami pola distribusi, prevalensi dan cara penul...

  13. Metallic mercury uptake by catalase Part 1 In Vitro metallic mercury uptake by various kind of animals' erythrocytes and purified human erythrocyte catalase

    OpenAIRE

    劒持,堅志

    1980-01-01

    The uptake of metallic mercury was studied using erythrocytes with different catalase activities taken from various kind of animals. The results were: 1) The uptake of metallic mercury by erythrocytes paralleled the activity of catalase in the erythrocytes with and without hydrogen peroxide, suggesting that the erythrocyte catalase activity is related to the uptake of metallic mercury. 2) The uptake of metallic mercury occurred not only with purified human erythrocyte catalase but also with h...

  14. Identification and subcellular localization of a 21-kilodalton molecule using affinity-purified antibodies against α-transforming growth factor

    International Nuclear Information System (INIS)

    Hazarika, P.; Pardue, R.L.; Earls, R.; Dedman, J.R.

    1987-01-01

    Monospecific antibodies were generated against each of six different peptide sequences derived from rat and human α-transforming growth factor (α-TGF). The affinity-purified antibody to the 17 amino acid carboxyl-terminal portion of the molecule proved most useful in detecting α-TGF. When used in a peptide-based radioimmunoassay, it was possible to measure nanogram quantities of native α-TGF in conditioned cell culture media. When used to analyze cell lysate, these antibodies specifically recognized a 21-kilodalton protein species. Indirect immunofluorescence localization procedures revealed a high concentration of α-TCF in a perinuclear ring with a diffuse cytoplasmic distribution. These results suggest that a precursor form of α-TGF has a cellular role beyond that of an autocrine growth factor

  15. Analyses of residual iron in carbon nanotubes produced by camphor/ferrocene pyrolysis and purified by high temperature annealing

    Energy Technology Data Exchange (ETDEWEB)

    Antunes, E.F., E-mail: ericafa@las.inpe.br [Instituto Tecnologico de Aeronautica (ITA), Praca Marechal Eduardo Gomes, 50, CEP 12.228-900, Sao Jose dos Campos, SP (Brazil); Instituto Nacional de Pesquisas Espaciais (INPE), Av. dos Astronautas, 1758, CEP 12.227-010, Sao Jose dos Campos, SP (Brazil); Resende, V.G. de; Mengui, U.A. [Instituto Nacional de Pesquisas Espaciais (INPE), Av. dos Astronautas, 1758, CEP 12.227-010, Sao Jose dos Campos, SP (Brazil); Cunha, J.B.M. [Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Goncalves, 9500, CEP 91.501-970, Porto Alegre, RS (Brazil); Corat, E.J.; Massi, M. [Instituto Nacional de Pesquisas Espaciais (INPE), Av. dos Astronautas, 1758, CEP 12.227-010, Sao Jose dos Campos, SP (Brazil)

    2011-07-01

    A detailed analysis of iron-containing phases in multiwall carbon nanotube (MWCNT) powder was carried out. The MWCNTs were produced by camphor/ferrocene and purified by high temperature annealing in an oxygen-free atmosphere (N{sub 2} or VC). Thermogravimetric analysis, Moessbauer spectroscopy, X-ray diffraction and X-ray photoelectron spectroscopy enabled the evaluation of the residual iron in MWCNTs after purification. The VC treatments provided MWCNTs with a purity degree higher than 99%. Moreover, Raman spectroscopy revealed a significant improvement in graphitic ordering after thermal annealing. A brief description of the mechanism of iron removal was included. We highlight the mobility of iron atoms through graphitic sheets and the large contact angle of iron clusters formed on MWCNT surfaces at high temperatures.

  16. An Artificial Channel Experiment for Purifying Drainage Water Containing Arsenic by Using Eleocharis acicularis

    Science.gov (United States)

    Okazaki, Kenji; Yamazaki, Shusaku; Kurahashi, Toshiyuki; Sakakibara, Masayuki

    2017-06-01

    This paper reports the results of an artificial channel experiment in which water containing arsenic was purified by using Eleocharis acicularis. The experiment was conducted to investigate the feasibility of phytoremediation by Eleocharis acicularis in civil engineering projects. In the experiment, 15 m2 of Eleocharis acicularis mats were laid in an artificial channel. Three sessions of artificial flow were implemented by leading 100.0 L of river water containing 0.234 mg/L of arsenic into the channel each time. The arsenic concentration of the leachate from the channel was analyzed. As the results of experiment, the arsenic concentrations of the leachate for the three sessions were 0.045 mg/L, 0.133 mg/L, and 0.249 mg/L. This shows that the arsenic concentration decreased during the first two sessions, whose flow totaled 200 L. The arsenic concentrations in the Eleocharis acicularis were 0.87 mg/kg, 1.01 mg/kg, and 4.16 mg/kg, which show that the plant absorbs arsenic. Moreover, it was found that the amount of sample water was reduced through evapotranspiration from the plant and the artificial channel.

  17. Hemolytic and cytotoxic properties of saponin purified from Holothuria leucospilota sea cucumber.

    Science.gov (United States)

    Soltani, Mozhgan; Parivar, Kazem; Baharara, Javad; Kerachian, Mohammad Amin; Asili, Javad

    2014-10-01

    Holothuroids (sea cucumbers) are members of the phylum echinodermata, which produce saponins. Saponins exhibit a wide spectrum of pharmacological and biological activities. In this study, we isolated the crude saponins from the body wall of the dominant Iranian species of sea cucumber, Holothuria leucospilota (H. leucospilota). The purpose of this study was to confirm the presence of saponins in the Persian Gulf H. leucospilota and study the hemolytic and cytotoxic activities of these compounds. The body wall of sea cucumber was dried and powdered and the crude saponins were isolated using various solvents. The crude saponins were further purified by column chromatography using HP-20 resin. The foam test, Thin Layer Chromatography (TLC), hemolytic assay, and Fourier Transform Infrared Spectroscopy (FTIR) confirmed the presence of saponins. Cytotoxicity was analyzed using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay on A549 cells, a human lung cancer cell line. The foam test, hemolytic assay, and TLC supported the presence of saponin compounds in the 80% ethanol fraction of H. leucospilota. The infrared (IR) spectrum of the extract showed hydroxyl (-OH), alkyl (C-H), ether (C-O) and ester (-C=O) absorption characteristic of teriterpenoid saponins. The C-O-C absorption indicated glycoside linkages to the sapogenins. The crude saponin extracted from sea cucumber was cytotoxic to A549 cells. The 80% ethanol fraction of saponin isolated from H. leucospilota exhibited hemolytic activity and offers promise as an anti-cancer candidate.

  18. N-terminal processing of affinity-tagged recombinant proteins purified by IMAC procedures.

    Science.gov (United States)

    Mooney, Jane T; Fredericks, Dale P; Christensen, Thorkild; Bruun Schiødt, Christine; Hearn, Milton T W

    2015-07-01

    The ability of a new class of metal binding tags to facilitate the purification of recombinant proteins, exemplified by the tagged glutathione S-transferase and human growth hormone, from Escherichia coli fermentation broths and lysates has been further investigated. These histidine-containing tags exhibit high affinity for borderline metal ions chelated to the immobilised ligand, 1,4,7-triazacyclononane (tacn). The use of this tag-tacn immobilised metal ion affinity chromatography (IMAC) system engenders high selectivity with regard to host cell protein removal and permits facile tag removal from the E. coli-expressed recombinant protein. In particular, these tags were specifically designed to enable their efficient removal by the dipeptidyl aminopeptidase 1 (DAP-1), thus capturing the advantages of high substrate specificity and rates of cleavage. MALDI-TOF MS analysis of the cleaved products from the DAP-1 digestion of the recombinant N-terminally tagged proteins confirmed the complete removal of the tag within 4-12 h under mild experimental conditions. Overall, this study demonstrates that the use of tags specifically designed to target tacn-based IMAC resins offers a comprehensive and flexible approach for the purification of E. coli-expressed recombinant proteins, where complete removal of the tag is an essential prerequisite for subsequent application of the purified native proteins in studies aimed at delineating the molecular and cellular basis of specific biological processes. Copyright © 2015 John Wiley & Sons, Ltd.

  19. Morphology and Molecular Composition of Purified Bovine Viral Diarrhea Virus Envelope.

    Directory of Open Access Journals (Sweden)

    Nathalie Callens

    2016-03-01

    Full Text Available The family Flaviviridae includes viruses that have different virion structures and morphogenesis mechanisms. Most cellular and molecular studies have been so far performed with viruses of the Hepacivirus and Flavivirus genera. Here, we studied bovine viral diarrhea virus (BVDV, a member of the Pestivirus genus. We set up a method to purify BVDV virions and analyzed their morphology by electron microscopy and their protein and lipid composition by mass spectrometry. Cryo-electron microscopy showed near spherical viral particles displaying an electron-dense capsid surrounded by a phospholipid bilayer with no visible spikes. Most particles had a diameter of 50 nm and about 2% were larger with a diameter of up to 65 nm, suggesting some size flexibility during BVDV morphogenesis. Morphological and biochemical data suggested a low envelope glycoprotein content of BVDV particles, E1 and E2 being apparently less abundant than Erns. Lipid content of BVDV particles displayed a ~2.3 to 3.5-fold enrichment in cholesterol, sphingomyelin and hexosyl-ceramide, concomitant with a 1.5 to 5-fold reduction of all glycerophospholipid classes, as compared to lipid content of MDBK cells. Although BVDV buds in the endoplasmic reticulum, its lipid content differs from a typical endoplasmic reticulum membrane composition. This suggests that BVDV morphogenesis includes a mechanism of lipid sorting. Functional analyses confirmed the importance of cholesterol and sphingomyelin for BVDV entry. Surprisingly, despite a high cholesterol and sphingolipid content of BVDV envelope, E2 was not found in detergent-resistant membranes. Our results indicate that there are differences between the structure and molecular composition of viral particles of Flaviviruses, Pestiviruses and Hepaciviruses within the Flaviviridae family.

  20. Partial oxidation process for producing a stream of hot purified gas

    Science.gov (United States)

    Leininger, T.F.; Robin, A.M.; Wolfenbarger, J.K.; Suggitt, R.M.

    1995-03-28

    A partial oxidation process is described for the production of a stream of hot clean gas substantially free from particulate matter, ammonia, alkali metal compounds, halides and sulfur-containing gas for use as synthesis gas, reducing gas, or fuel gas. A hydrocarbonaceous fuel comprising a solid carbonaceous fuel with or without liquid hydrocarbonaceous fuel or gaseous hydrocarbon fuel, wherein said hydrocarbonaceous fuel contains halides, alkali metal compounds, sulfur, nitrogen and inorganic ash containing components, is reacted in a gasifier by partial oxidation to produce a hot raw gas stream comprising H{sub 2}, CO, CO{sub 2}, H{sub 2}O, CH{sub 4}, NH{sub 3}, HCl, HF, H{sub 2}S, COS, N{sub 2}, Ar, particulate matter, vapor phase alkali metal compounds, and molten slag. The hot raw gas stream from the gasifier is split into two streams which are separately deslagged, cleaned and recombined. Ammonia in the gas mixture is catalytically disproportionated into N{sub 2} and H{sub 2}. The ammonia-free gas stream is then cooled and halides in the gas stream are reacted with a supplementary alkali metal compound to remove HCl and HF. Alkali metal halides, vaporized alkali metal compounds and residual fine particulate matter are removed from the gas stream by further cooling and filtering. The sulfur-containing gases in the process gas stream are then reacted at high temperature with a regenerable sulfur-reactive mixed metal oxide sulfur sorbent material to produce a sulfided sorbent material which is then separated from the hot clean purified gas stream having a temperature of at least 1000 F. 1 figure.

  1. Optoacoustic tomography in preclinical research: in vivo distribution of highly purified PEG-coated gold nanorods

    Science.gov (United States)

    Su, Richard; Liopo, Anton; Brecht, Hans-Peter; Ermilov, Sergey; Larin, Kirill; Oraevsky, Alexander A.

    2011-07-01

    We report on the optoacoustic (OA) imaging of the whole mouse body using a biocompatible contrast agent - highly purified, pegylated gold nanorods (GNR) - which has strong optical absorption in the near-infrared region and low level of toxicity. In vitro toxicity studies showed no significant change in survival rates of the cultured normal epithelium IEC-6 cells when incubated for 24 hours with up to 1 nM of GNR. In vivo toxicity studies in nude mice showed no pathological changes in liver 1 month after the IV injection of GNR with intra-body concentration around 0.25-0.50 nM. In order to study the enhancement of the OA contrast and accumulation of GNR in different tissues, we performed 3D OA imaging of live nude mice with IV-injected GNR. The enhancement of the OA contrast in comparison with the images of the untreated mice was visible starting 1 hour after the GNR injection. The OA contrast of kidneys, liver, and spleen peaked at about 2-3 days after the administration of the GNR, and then was gradually reducing.

  2. Anti-Taenia solium metacestodes antibodies in serum from blood donors from four cities of Triângulo Mineiro area, Minas Gerais, Brazil, 1995 Anticorpos anti-formas metacestódeas de Taenia solium em amostras de soros de doadores de sangue de quatro cidades da região do Triângulo Mineiro, Minas Gerais, Brasil, 1995

    Directory of Open Access Journals (Sweden)

    Elisângela de Paula SILVEIRA-LACERDA

    2002-07-01

    Full Text Available Serological survey was performed to detect IgG antibodies anti-Taenia solium metacestodes in blood donors of Hemocentro Regional de Uberlândia, Minas Gerais, Brazil. A total of 1133 sera from blood donors coming from four cities of Triângulo Mineiro area were analyzed by the indirect fluorescence antibody test (IFAT and the enzyme linked immunosorbent assay (ELISA. Specific IgG antibodies were found in 5.6% of the studied population, showing differences in the positive rates according to their origin: Araguari (13.5%, Tupaciguara (5.0%, Monte Alegre de Minas (4.8% and Uberlândia (4.7%. The results indicate the probable endemicity of cysticercosis in this population.Realizou-se pesquisa sorológica para detectar anticorpos IgG anti-formas metacestódeas de Taenia solium em doadores de sangue do Hemocentro Regional de Uberlândia, Minas Gerais, Brasil. O total de 1133 amostras de soros de doadores de sangue de quatro cidades do Triângulo Mineiro foi analisado pelo teste de imunofluorescência indireta (IFI e o teste imunoenzimático (ELISA. Anticorpos IgG específicos foram detectados em 5,6% da população estudada, mostrando diferenças nas taxas de positividade de acordo com suas cidades de origens: Araguari (13,5%, Tupaciguara (5,0%, Monte Alegre de Minas (4,8% e Uberlândia (4,7%. Os resultados indicam a provável endemicidade de cisticercose nesta população.

  3. Studies on cell-free metabolism: ethanol production by a yeast glycolytic system reconstituted from purified enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Welch, P; Scopes, R K

    1985-07-01

    A reconstituted glycolytic system has been established from individually purified enzymes to simulate the conversion of glucose to ethanol plus CO/sub 2/ by yeast. Sustained and extensive conversion occurred provided that input of glucose matched the rate of ATP degradation appropriately. ATPase activity could be replaced by arsenate, which uncoupled ATP synthesis from glycolysis. The mode of uncoupling was investigated, and it was concluded that the artificial intermediate, 1-arseno-3-phosphoglycerate, has a half-life of no more than a few milliseconds. Arsenate at 4 mM concentration could simulate the equivalent of 10 ..mu..mol/ml min. of ATPase activity. The reconstituted enzyme system was capable of totally degrading one M (18% w/v) glucose in 8 hours giving 9% (w/v) ethanol. The levels of metabolites during metabolism were measured to detect rate-limiting steps. The successful operation of the reconstituted enzyme system demonstrates that it is possible to carry out complex chemical transformations with multiple enzyme systems in vitro. 36 references.

  4. Comparison of safety and immunogenicity of purified chick embryo cell vaccine using Zagreb and Essen regimens in patients with category II exposure in China.

    Science.gov (United States)

    Hu, Quan; Liu, Man-Qing; Zhu, Zheng-Gang; Zhu, Ze-Rong; Lu, Sha

    2014-01-01

    The aim was to compare the safety and immunogenicity of purified chick embryo cell vaccine (PCECV) with Zagreb 2-1-1 and Essen 1-1-1-1-1 regimens in patients with WHO category II exposure in China. Side effects including systemic and local symptoms were recorded for all patients during vaccination with purified chick embryo cell vaccine (PCECV) under Zagreb 2-1-1 or Essen 1-1-1-1-1 regimens, and the rabies neutralization antibody titers in patients' serum at days 0, 7, 14, 45, 365 post-immunization were measured to determine the immunogenicity. Fever and pain were the most common events for systemic and local symptoms respectively, and most side effects (86.78%, 105/121) occurred after the first dose of vaccination. Safety analysis showed differences in side effects inZagreb and Essen regimens, especially after the first dose of vaccination (P = 0.043). Immunogenicity analysis indicated that Zagreb can achieve higher neutralization antibody titers and a greater seroconversion rate in a shorter time but had less persistence than Essen. When compared with the Essen regimen, the Zagreb regimen had a different immunogenicity in all study subjects, and different safety profile in young children, and a further study with a larger population and longer surveillance is warranted.

  5. Discovery and Targeted LC-MS/MS of Purified Polerovirus Reveals Differences in the Virus-Host Interactome Associated with Altered Aphid Transmission

    Science.gov (United States)

    Howe, Kevin; Fish, Tara; Smith, Dawn; Gildow, Fredrick; MacCoss, Michael J.; Thannhauser, Theodore W.; Gray, Stewart M.

    2012-01-01

    Circulative transmission of viruses in the Luteoviridae, such as cereal yellow dwarf virus (CYDV), requires a series of precisely orchestrated interactions between virus, plant, and aphid proteins. Natural selection has favored these viruses to be retained in the phloem to facilitate acquisition and transmission by aphids. We show that treatment of infected oat tissue homogenate with sodium sulfite reduces transmission of the purified virus by aphids. Transmission electron microscopy data indicated no gross change in virion morphology due to treatments. However, treated virions were not acquired by aphids through the hindgut epithelial cells and were not transmitted when injected directly into the hemocoel. Analysis of virus preparations using nanoflow liquid chromatography coupled to tandem mass spectrometry revealed a number of host plant proteins co-purifying with viruses, some of which were lost following sodium sulfite treatment. Using targeted mass spectrometry, we show data suggesting that several of the virus-associated host plant proteins accumulated to higher levels in aphids that were fed on CYDV-infected plants compared to healthy plants. We propose two hypotheses to explain these observations, and these are not mutually exclusive: (a) that sodium sulfite treatment disrupts critical virion-host protein interactions required for aphid transmission, or (b) that host infection with CYDV modulates phloem protein expression in a way that is favorable for virus uptake by aphids. Importantly, the genes coding for the plant proteins associated with virus may be examined as targets in breeding cereal crops for new modes of virus resistance that disrupt phloem-virus or aphid-virus interactions. PMID:23118947

  6. Discovery and targeted LC-MS/MS of purified polerovirus reveals differences in the virus-host interactome associated with altered aphid transmission.

    Directory of Open Access Journals (Sweden)

    Michelle Cilia

    Full Text Available Circulative transmission of viruses in the Luteoviridae, such as cereal yellow dwarf virus (CYDV, requires a series of precisely orchestrated interactions between virus, plant, and aphid proteins. Natural selection has favored these viruses to be retained in the phloem to facilitate acquisition and transmission by aphids. We show that treatment of infected oat tissue homogenate with sodium sulfite reduces transmission of the purified virus by aphids. Transmission electron microscopy data indicated no gross change in virion morphology due to treatments. However, treated virions were not acquired by aphids through the hindgut epithelial cells and were not transmitted when injected directly into the hemocoel. Analysis of virus preparations using nanoflow liquid chromatography coupled to tandem mass spectrometry revealed a number of host plant proteins co-purifying with viruses, some of which were lost following sodium sulfite treatment. Using targeted mass spectrometry, we show data suggesting that several of the virus-associated host plant proteins accumulated to higher levels in aphids that were fed on CYDV-infected plants compared to healthy plants. We propose two hypotheses to explain these observations, and these are not mutually exclusive: (a that sodium sulfite treatment disrupts critical virion-host protein interactions required for aphid transmission, or (b that host infection with CYDV modulates phloem protein expression in a way that is favorable for virus uptake by aphids. Importantly, the genes coding for the plant proteins associated with virus may be examined as targets in breeding cereal crops for new modes of virus resistance that disrupt phloem-virus or aphid-virus interactions.

  7. Discovery and targeted LC-MS/MS of purified polerovirus reveals differences in the virus-host interactome associated with altered aphid transmission.

    Science.gov (United States)

    Cilia, Michelle; Peter, Kari A; Bereman, Michael S; Howe, Kevin; Fish, Tara; Smith, Dawn; Gildow, Fredrick; MacCoss, Michael J; Thannhauser, Theodore W; Gray, Stewart M

    2012-01-01

    Circulative transmission of viruses in the Luteoviridae, such as cereal yellow dwarf virus (CYDV), requires a series of precisely orchestrated interactions between virus, plant, and aphid proteins. Natural selection has favored these viruses to be retained in the phloem to facilitate acquisition and transmission by aphids. We show that treatment of infected oat tissue homogenate with sodium sulfite reduces transmission of the purified virus by aphids. Transmission electron microscopy data indicated no gross change in virion morphology due to treatments. However, treated virions were not acquired by aphids through the hindgut epithelial cells and were not transmitted when injected directly into the hemocoel. Analysis of virus preparations using nanoflow liquid chromatography coupled to tandem mass spectrometry revealed a number of host plant proteins co-purifying with viruses, some of which were lost following sodium sulfite treatment. Using targeted mass spectrometry, we show data suggesting that several of the virus-associated host plant proteins accumulated to higher levels in aphids that were fed on CYDV-infected plants compared to healthy plants. We propose two hypotheses to explain these observations, and these are not mutually exclusive: (a) that sodium sulfite treatment disrupts critical virion-host protein interactions required for aphid transmission, or (b) that host infection with CYDV modulates phloem protein expression in a way that is favorable for virus uptake by aphids. Importantly, the genes coding for the plant proteins associated with virus may be examined as targets in breeding cereal crops for new modes of virus resistance that disrupt phloem-virus or aphid-virus interactions.

  8. PRODUCTION AND PURIFICATION OF IgY ANTIBODIES AS A NOVEL TOOL TO PURIFY THE NR1 SUBUNIT OF NMDA RECEPTO

    Directory of Open Access Journals (Sweden)

    Edgar Antonio Reyes Montaño

    2011-12-01

    Full Text Available Producing polyclonal antibodies (IgY inchickens has advantages over those obtainedin other animal models, since theyhave been used as a tool for studyingdifferent proteins (NMDA glutamate receptorin our case, specifically the NR1subunit. We produced specific antibodiesagainst expression products by thealternative splicing of the gene encodingNMDA receptor NR1 subunit in adult ratbrain. Three peptides corresponding tothe splicing sites (N1, C1 and C2’ cassetteswere designed, synthesised and usedindividually as antigens in hens. Specificimmunoglobulins were purified fromyolks. The antibodies were then used forpurifying the NMDA receptor NR1 subunitusing affinity chromatography couplingthe three antibodies to the support.R

  9. Comparative study of two purified inulinases from thermophile Thielavia Terrestris NRRL 8126 and mesophile Aspergillus Foetidus NRRL 337 grown on Cichorium Intybus l

    Directory of Open Access Journals (Sweden)

    Eman Mohamed Fawzi

    2011-06-01

    Full Text Available Thirty fungal species grown on Cichorium intybus L. root extract as a sole carbon source, were screened for the production of exo-inulinase activities. The thermophile Thielavia terrestris NRRL 8126 and mesophile Aspergillus foetidus NRRL 337 gave the highest production levels of inulinases I & II at 50 and 24 ºC respectively. Yeast extract and peptone were the best nitrogen sources for highest production of inulinases I & II at five and seven days of incubation respectively. The two inulinases I & II were purified to homogeneity by gel-filtration and ion-exchange chromatography with 66.0 and 42.0 fold of purification respectively. The optimum temperatures of purified inulinases I & II were 75 and 50 ºC respectively. Inulinase I was more thermostable than the other one. The optimum pH for activity was found to be 4.5 and 5.5 for inulinases I & II respectively. A comparatively lower Michaelis-Menten constant (2.15 mg/ml and higher maximum initial velocity (115 µmol/min/mg of protein for inulinase I on inulin demonstrated the exoinulinase's greater affinity for inulin substrate. These findings are significant for its potential industrial application. The molecular mass of the inulinases I & II were estimated to be 72 & 78 kDa respectively by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

  10. Structure/activity relationship of thapsigargin inhibition on the purified Golgi/secretory pathway Ca2+/Mn2+-transport ATPase (SPCA1a)

    DEFF Research Database (Denmark)

    Chen, Jialin; De Raeymaecker, Joren; Hovgaard, Jannik Brondsted

    2017-01-01

    SPCA1a displays a higher apparent Ca2+ affinity and lower maximal turnover rate than the purified sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA1a). The lipids cholesteryl hemisuccinate, linole-/oleamide and phosphatidyl ethanolamine inhibit, whereas phosphatidic acid and sphingomyelin enhance SPCA1a...... activity. Moreover, SPCA1a is blocked by μM concentrations of commonly used SERCA1a inhibitors thapsigargin (Tg), cyclopiazonic acid (CPA) and 2,5-di-tert-butyl hydroquinone (BHQ). Since tissue-specific targeting of SERCA2b by Tg analogues is considered for prostate cancer therapy, the inhibition of SPCA1a...

  11. Structural Identification of 19 Purified Isomers of the OPV Acceptor Material bisPCBM by 13C NMR and UV-Vis Absorption Spectroscopy and High-Performance Liquid Chromatography.

    Science.gov (United States)

    Liu, Tong; Abrahams, Isaac; Dennis, T John S

    2018-04-26

    The molecular structures of 19 purified isomers of bis-phenyl-C 62 -butyric acid methyl ester were identified by a combination of 13 C NMR and UV-vis absorption spectroscopies and high-performance liquid chromatography (HPLC) retention time analysis. All 19 isomers are dicyclopropafullerenes (none are homofullerenes). There were seven isomers with C 1 molecular point-group symmetry, four with C s , six with C 2 , one with C 2 v , and one with C 2 h symmetry. The C 2 h , C 2 v , and all five nonequatorial C 1 isomers were unambiguously assigned to their respective HPLC fractions. For the other 12 isomers, the 13 C NMR and UV-vis spectra placed them in six groups of two same-symmetry isomers. On the basis of the widely spaced HPLC retention times of the two isomers within each of these six groups, and the empirical inverse correlation between retention time and addend spacing, each isomer was assigned to its corresponding HPLC fraction. In addition, the missing trans-1 isomer was found, purified, and characterized.

  12. Sensitization to epithelial antigens in chronic mucosal inflammatory disease. Characterization of human intestinal mucosa-derived mononuclear cells reactive with purified epithelial cell-associated components in vitro.

    OpenAIRE

    Roche, J K; Fiocchi, C; Youngman, K

    1985-01-01

    To explore the auto-reactive potential of cells infiltrating the gut mucosa in idiopathic chronic inflammatory bowel disease, intestinal lamina propria mononuclear cells (LPMC) were isolated, characterized morphologically and phenotypically, and evaluated for antigen-specific reactivity. The last was assessed by quantitating LPMC cytotoxic capabilities against purified, aqueous-soluble, organ-specific epithelial cell-associated components (ECAC) characterized previously. Enzyme-isolated infla...

  13. Characterization of purified and Xerogel immobilized Novel Lignin Peroxidase produced from Trametes versicolor IBL-04 using solid state medium of Corncobs

    Directory of Open Access Journals (Sweden)

    Asgher Muhammad

    2012-08-01

    Full Text Available Abstract Background Cost-effective production of industrially important enzymes is a key for their successful exploitation on industrial scale. Keeping in view the extensive industrial applications of lignin peroxidase (LiP, this study was performed to purify and characterize the LiP from an indigenous strain of Trametes versicolor IBL-04. Xerogel matrix enzyme immobilization technique was applied to improve the kinetic and thermo-stability characteristics of LiP to fulfil the requirements of the modern enzyme consumer sector of biotechnology. Results A novel LiP was isolated from an indigenous T. versicolor IBL-04 strain. T. versicolor IBL-04 was cultured in solid state fermentation (SSF medium of corn cobs and maximum LiP activity of 592 ± 6 U/mL was recorded after five days of incubation under optimum culture conditions. The crude LiP was 3.3-fold purified with specific activity of 553 U/mg after passing through the DEAE-cellulose and Sephadex-G-100 chromatography columns. The purified LiP exhibited a relatively low molecular weight (30 kDa homogenous single band on native and SDS-PAGE. The LiP was immobilized by entrapping in xerogel matrix of trimethoxysilane (TMOS and proplytetramethoxysilane (PTMS and maximum immobilization efficiency of 88.6% was achieved. The free and immobilized LiPs were characterized and the results showed that the free and immobilized LiPs had optimum pH 6 and 5 while optimum temperatures were 60°C and 80°C, respectively. Immobilization was found to enhance the activity and thermo-stability potential of LiP significantly and immobilized LiP remained stable over broad pH and temperature range as compare to free enzyme. Kinetic constants Km and Vmax were 70 and 56 μM and 588 and 417 U/mg for the free and immobilized LiPs, respectively. Activity of this novel extra thermo-stable LiP was stimulated to variable extents by Cu2+, Mn2+ and Fe2+ whereas, Cystein, EDTA and Ag+ showed inhibitory effects

  14. Decontamination efficiencies of pot-type water purifiers for ¹³¹I, ¹³⁴Cs and ¹³⁷Cs in rainwater contaminated during Fukushima Daiichi nuclear disaster.

    Directory of Open Access Journals (Sweden)

    Shogo Higaki

    Full Text Available Rainwater was contaminated by a large release of radionuclides into the environment during the Fukushima Daiichi nuclear disaster. It became a matter of concern for Japan when several water purification plants detected ¹³¹I contamination in the drinking water. In the present study, the decontamination efficiency of two easily obtainable commercial water purifiers were examined for rainwater contaminated with ¹³¹I, ¹³⁴Cs and ¹³⁷Cs. The water purifiers removed 94.2-97.8% of the ¹³¹I and 84.2-91.5% of the ¹³⁴Cs and ¹³⁷Cs after one filtration. Seven filtrations removed 98.2-99.6% of the ¹³¹I and over 98.0% of the ¹³⁴Cs and ¹³⁷Cs. From a practical perspective, over the fourth filtrations were not needed because of no significant improvements after the third filtration.

  15. Hemolytic and Cytotoxic Properties of Saponin Purified from Holothuria leucospilota Sea Cucumber

    Directory of Open Access Journals (Sweden)

    Mozhgan Soltani

    2014-10-01

    Full Text Available Background: Holothuroids (sea cucumbers are members of the phylum echinodermata, which produce saponins. Saponins exhibit a wide spectrum of pharmacological and biological activities. In this study, we isolated the crude saponins from the body wall of the dominant Iranian species of sea cucumber, Holothuria leucospilota (H. leucospilota. The purpose of this study was to confirm the presence of saponins in the Persian Gulf H. leucospilota and study the hemolytic and cytotoxic activities of these compounds. Methods: The body wall of sea cucumber was dried and powdered and the crude saponins were isolated using various solvents. The crude saponins were further purified by column chromatography using HP-20 resin. The foam test, Thin Layer Chromatography (TLC, hemolytic assay, and Fourier Transform Infrared Spectroscopy (FTIR confirmed the presence of saponins. Cytotoxicity was analyzed using a 3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide (MTT assay on A549 cells, a human lung cancer cell line. Results: The foam test, hemolytic assay, and TLC supported the presence of saponin compounds in the 80% ethanol fraction of H. leucospilota. The infrared (IR spectrum of the extract showed hydroxyl (-OH, alkyl (C-H, ether (C-O and ester (–C=O absorption characteristic of teriterpenoid saponins. The C-O-C absorption indicated glycoside linkages to the sapogenins. The crude saponin extracted from sea cucumber was cytotoxic to A549 cells. Conclusion: The 80% ethanol fraction of saponin isolated from H. leucospilota exhibited hemolytic activity and offers promise as an anti-cancer candidate.

  16. Evaluation of anti-acne property of purified bee venom serum in humans.

    Science.gov (United States)

    Han, Sang Mi; Pak, Sok Cheon; Nicholls, Young Mee; Macfarlane, Nicola

    2016-12-01

    Acne vulgaris is a chronic dermatologic disease with four factors involved in the development of lesions. Treatments need to address as many of these underlying factors as possible in order to reduce acne lesions. As such, purified bee venom (PBV ™ ) serum is an attractive therapeutic option for acne, but little data exist on the efficacy of this treatment strategy. In this prospective, noncomparative study, 30 subjects having mild-to-moderate acne vulgaris were enrolled and treated with PBV ™ serum twice daily for a period of 6 weeks. Clinical evaluation of lesions by expert visual grading and image analysis were made at weeks 0 (baseline), 3, and 6. The average visual acne grade of all volunteers significantly improved with the PBV ™ serum treatment at weeks 3 (P < 0.05) and 6 (P < 0.001) when compared with the baseline grade at week 0. In addition, there was a mean percent improvement of 8.6% and 52.3% in acne grade observed after 3 and 6 weeks of PBV ™ serum use, with 20% and 77% of the subjects showing improvement, respectively, when compared with baseline. Moreover, the subjects showed improvement in open comedones, closed comedones, papules, pustules, and nodules after 3 and 6 weeks of PBV ™ serum use. Six weeks of treatment with PBV ™ serum was found to be effective in the treatment of mild-to-moderate acne vulgaris, with no incidence of serious side effects or irritation. © 2016 The Authors Journal of Cosmetic Dermatology Published by Wiley Periodicals, Inc.

  17. Kinetic Characterisation of Phosphofructokinase Purified from Setaria cervi: A Bovine Filarial Parasite

    Directory of Open Access Journals (Sweden)

    Bechan Sharma

    2011-01-01

    Full Text Available Phosphofructokinase (PFK, a regulatory enzyme in glycolytic pathway, has been purified to electrophoretic homogeneity from adult female Setaria cervi and partially characterized. For this enzyme, the Lineweaver-Burk's double reciprocal plots of initial rates and D-fructose-6-phosphate (F-6-P or Mg-ATP concentrations for varying values of cosubstrate concentration gave intersecting lines indicating that Km values for F-6-P (1.05 mM and ATP (3 μM were independent of each other. S. cervi PFK, when assayed at inhibitory concentration of ATP (>0.1 mM, exhibited sigmoidal behavior towards binding with F-6-P with a Hill coefficient (n value equal to 1.8 and 1.7 at 1.0 and 0.33 mM ATP, respectively. D-fructose-1,6-diphosphate (FDP competitively inhibited the filarial enzyme: Ki and Hill coefficient values being 0.18 μM and 2.0, respectively. Phosphoenolpyruvate (PEP also inhibited the enzyme competitively with the Ki value equal to 0.8 mM. The Hill coefficient values (>1.5 for F-6-P (at inhibitory concentration of ATP and FDP suggested its positive cooperative kinetics towards F-6-P and FDP, showing presence of more than one binding sites for these molecules in enzyme protein and allosteric nature of the filarial enzyme. The product inhibition studies gave us the only compatible mechanism of random addition process with a probable orientation of substrates and products on the enzyme surface.

  18. Purified phenolics from hydrothermal treatments of biomass: ability to protect sunflower bulk oil and model food emulsions from oxidation.

    Science.gov (United States)

    Conde, Enma; Moure, Andrés; Domínguez, Herminia; Gordon, Michael H; Parajó, Juan Carlos

    2011-09-14

    The phenolic fractions released during hydrothermal treatment of selected feedstocks (corn cobs, eucalypt wood chips, almond shells, chestnut burs, and white grape pomace) were selectively recovered by extraction with ethyl acetate and washed with ethanol/water solutions. The crude extracts were purified by a relatively simple adsorption technique using a commercial polymeric, nonionic resin. Utilization of 96% ethanol as eluting agent resulted in 47.0-72.6% phenolic desorption, yielding refined products containing 49-60% w/w phenolics (corresponding to 30-58% enrichment with respect to the crude extracts). The refined extracts produced from grape pomace and from chestnut burs were suitable for protecting bulk oil and oil-in-water and water-in-oil emulsions. A synergistic action with bovine serum albumin in the emulsions was observed.

  19. Treatment of snake envenomations by a new polyvalent antivenom composed of highly purified F(ab')2 : results of a clinical trial in northern Cameroon

    OpenAIRE

    Chippaux, Jean-Philippe; Lang, J.; Amadi-Eddine, S.; Fagot, P.; Le Mener, V.

    1999-01-01

    A clinical trail was conducted in 2 health centers in northern Cameroon to assess the safety and efficacy of a new polyvalent antivenom composed of higly purified and pasteurized F(ab')2 (FAV-Africa). Forty-six patients with objective signs of envenomation, including 67% with hemorrhage, were included in the study. Each patient received at least 20 ml of FAV-Africa by direct, slow intravenous injection ; 172 10-ml ampules were administered. All patients were clinically cured after treatment. ...

  20. [Biomaterials for bone filling: comparisons between autograft, hydroxyapatite and one highly purified bovine xenograft].

    Science.gov (United States)

    Chappard, D; Zhioua, A; Grizon, F; Basle, M F; Rebel, A

    1993-12-01

    Bone grafts are becoming increasingly common in orthopaedics, neurosurgery and periodontology. Twenty one New Zealand rabbits were used in the present study comparing several materials usable as bone substitutes. A 4.5 mm hole was drilled in the inner femoral condyles. Holes were filled with either an autograft (from the opposite condyle), an hydroxylapatite (Bioapatite), or a highly purified bovine xenograft (T650 Lubboc). Animals were sacrificed at 1, 3 and 6 months post implantation and a quantitative analysis of newly-formed bone volume (BNF/IV) and remaining biomaterials (BMAT/IV) was done. In addition, some holes were left unfilled and served as controls. At 6 months, there was no tendency for spontaneous repair in the control animals. The autografted animals have repaired their trabecular mass and architecture within the first month. Hydroxylapatite appeared unresorbed at six months and only thin and scanty new trabeculae were observed. The xenograft induced woven bone trabeculae formation on the first month. This was associated with resorption of the material by two multinucleated cell populations. At six months, the epiphyseal architecture was restored and the biomaterial has disappeared in most cases. Xenografts appear a promising alternative to autografts and allografts, whose infectious risks and ethical problems should always be borne in mind.

  1. Site-directed mutagenesis in Petunia × hybrida protoplast system using direct delivery of purified recombinant Cas9 ribonucleoproteins.

    Science.gov (United States)

    Subburaj, Saminathan; Chung, Sung Jin; Lee, Choongil; Ryu, Seuk-Min; Kim, Duk Hyoung; Kim, Jin-Soo; Bae, Sangsu; Lee, Geung-Joo

    2016-07-01

    Site-directed mutagenesis of nitrate reductase genes using direct delivery of purified Cas9 protein preassembled with guide RNA produces mutations efficiently in Petunia × hybrida protoplast system. The clustered, regularly interspaced, short palindromic repeat (CRISPR)-CRISPR associated endonuclease 9 (CRISPR/Cas9) system has been recently announced as a powerful molecular breeding tool for site-directed mutagenesis in higher plants. Here, we report a site-directed mutagenesis method targeting Petunia nitrate reductase (NR) gene locus. This method could create mutations efficiently using direct delivery of purified Cas9 protein and single guide RNA (sgRNA) into protoplast cells. After transient introduction of RNA-guided endonuclease (RGEN) ribonucleoproteins (RNPs) with different sgRNAs targeting NR genes, mutagenesis at the targeted loci was detected by T7E1 assay and confirmed by targeted deep sequencing. T7E1 assay showed that RGEN RNPs induced site-specific mutations at frequencies ranging from 2.4 to 21 % at four different sites (NR1, 2, 4 and 6) in the PhNR gene locus with average mutation efficiency of 14.9 ± 2.2 %. Targeted deep DNA sequencing revealed mutation rates of 5.3-17.8 % with average mutation rate of 11.5 ± 2 % at the same NR gene target sites in DNA fragments of analyzed protoplast transfectants. Further analysis from targeted deep sequencing showed that the average ratio of deletion to insertion produced collectively by the four NR-RGEN target sites (NR1, 2, 4, and 6) was about 63:37. Our results demonstrated that direct delivery of RGEN RNPs into protoplast cells of Petunia can be exploited as an efficient tool for site-directed mutagenesis of genes or genome editing in plant systems.

  2. Simple Identification of Human Taenia Species by Multiplex Loop-Mediated Isothermal Amplification in Combination with Dot Enzyme-Linked Immunosorbent Assay.

    Science.gov (United States)

    Nkouawa, Agathe; Sako, Yasuhito; Okamoto, Munehiro; Ito, Akira

    2016-06-01

    For differential detection of Taenia solium, Taenia saginata, and Taenia asiatica, loop-mediated isothermal amplification (LAMP) assay targeting the cytochrome c oxidase subunit 1 gene has been recently developed and shown to be sensitive, specific, and effective. However, to achieve differential identification, one specimen requires three reaction mixtures containing a primer set of each Taenia species separately, which is complex and time consuming and increases the risk of cross-contamination. In this study, we developed a simple differential identification of human Taenia species using multiplex LAMP (mLAMP) in combination with dot enzyme-linked immunosorbent assay (dot-ELISA). Forward inner primers of T. solium, T. saginata, and T. asiatica labeled with fluorescein isothiocyanate (FITC), digoxigenin (DIG), and tetramethylrhodamine (TAMRA), respectively, and biotin-labeled backward inner primers were used in mLAMP. The mLAMP assay succeeded in specific amplification of each respective target gene in a single tube. Furthermore, the mLAMP product from each species was easily distinguished by dot-ELISA with an antibody specific for FITC, DIG, or TAMRA. The mLAMP assay in combination with dot-ELISA will make identification of human Taenia species simpler, easier, and more practical. © The American Society of Tropical Medicine and Hygiene.

  3. General aspects concerning strictly meat and fish transmitted parasitic infections

    Directory of Open Access Journals (Sweden)

    Daniele Crotti

    2012-03-01

    Full Text Available All helminths parasitosis transmitted to humans trough ingestion of infested fleshes, where man is definitive host too, are represented by four groups of helminths: the cestodes Dyphyllobothrium spp and Spirometra spp. (Sparganum proliferum is the name of the immature plerocercoid larva, the trematodes Opisthorchis Clonorchis “group” (many could be the genera and species involved, and the nematode Capillaria philippinensis. So, for fishes humans foods (fresh or salted water the control and prevention in veterinary health must be directed to investigation regarding intermediate stages of these parasites in fishes for human alimentation; if present, they must be eliminated. The helminths parasitosis transmitted to humans trough ingestion of infected mammals meats, are represented by taeniasis (Taenia saginata, T. solium and T. saginata asiatica, where man id definitive host and the infection is caused by ingestion of bovine or swine meat, containing larvae of these cestodes, and by trichinellosis, where humans represent a intermediate stage, and the eventual pathology is caused as by adult (acute infection as by larvae (chronic infection of this nematode: usually the meats responsible are infected pork, wild pork or horse (Trichinella spp. Is inside the meats of these animals. So the veterinary control and prophylaxis are necessary to avoid this disease and preventing the infection that could be severe.

  4. Comparison of the Effect of Curing Ingredients Derived from Purified and Natural Sources on Inhibition of Clostridium perfringens Outgrowth during Cooling of Deli-Style Turkey Breast.

    Science.gov (United States)

    King, Amanda M; Glass, Kathleen A; Milkowski, Andrew L; Sindelar, Jeffrey J

    2015-08-01

    The antimicrobial impact of purified and natural sources of both nitrite and ascorbate were evaluated against Clostridium perfringens during the postthermal processing cooling period of deli-style turkey breast. The objective of phase I was to assess comparable concentrations of nitrite (0 or 100 ppm) and ascorbate (0 or 547 ppm) from both purified and natural sources. Phase II was conducted to investigate concentrations of nitrite (50, 75, or 100 ppm) from cultured celery juice powder and ascorbate (0, 250, or 500 ppm) from cherry powder to simulate alternative curing formulations. Ground turkey breast (75% moisture, 1.2% salt, pH 6.2) treatments were inoculated with C. perfringens spores (three-strain mixture) to yield 2.5 log CFU/g. Individual 50-g portions were vacuum packaged, cooked to 71.1°C, and chilled from 54.4 to 26.7°C in 5 h and from 26.7 to 7.2°C in 10 additional hours. Triplicate samples were assayed for growth of C. perfringens at predetermined intervals by plating on tryptose-sulfite-cycloserine agar; experiments were replicated three times. In phase I, uncured, purified nitrite, and natural nitrite treatments without ascorbate had 5.3-, 4.2-, and 4.4-log increases in C. perfringens, respectively, at 15 h, but nitrite and 547 ppm of ascorbate from either source. In phase II, 0, 50, 75, and 100 ppm of nitrite and 50 ppm of nitrite plus 250 ppm of ascorbate supported 4.5-, 3.9-, 3.5-, 2.2-, and 1.5-log increases in C. perfringens, respectively. In contrast, nitrite and 500 ppm of ascorbate or ≥75 ppm of nitrite and ≥250 ppm of ascorbate. These results confirm that equivalent concentrations of nitrite, regardless of the source, provide similar inhibition of C. perfringens during chilling and that ascorbate enhances the antimicrobial effect of nitrite on C. perfringens at concentrations commonly used in alternative cured meats.

  5. Multi-species sequence comparison reveals dynamic evolution of the elastin gene that has involved purifying selection and lineage-specific insertions/deletions

    Directory of Open Access Journals (Sweden)

    Green Eric D

    2004-05-01

    Full Text Available Abstract Background The elastin gene (ELN is implicated as a factor in both supravalvular aortic stenosis (SVAS and Williams Beuren Syndrome (WBS, two diseases involving pronounced complications in mental or physical development. Although the complete spectrum of functional roles of the processed gene product remains to be established, these roles are inferred to be analogous in human and mouse. This view is supported by genomic sequence comparison, in which there are no large-scale differences in the ~1.8 Mb sequence block encompassing the common region deleted in WBS, with the exception of an overall reversed physical orientation between human and mouse. Results Conserved synteny around ELN does not translate to a high level of conservation in the gene itself. In fact, ELN orthologs in mammals show more sequence divergence than expected for a gene with a critical role in development. The pattern of divergence is non-conventional due to an unusually high ratio of gaps to substitutions. Specifically, multi-sequence alignments of eight mammalian sequences reveal numerous non-aligning regions caused by species-specific insertions and deletions, in spite of the fact that the vast majority of aligning sites appear to be conserved and undergoing purifying selection. Conclusions The pattern of lineage-specific, in-frame insertions/deletions in the coding exons of ELN orthologous genes is unusual and has led to unique features of the gene in each lineage. These differences may indicate that the gene has a slightly different functional mechanism in mammalian lineages, or that the corresponding regions are functionally inert. Identified regions that undergo purifying selection reflect a functional importance associated with evolutionary pressure to retain those features.

  6. Fatty acid and drug binding to a low-affinity component of human serum albumin, purified by affinity chromatography

    DEFF Research Database (Denmark)

    Vorum, H; Pedersen, A O; Honoré, B

    1992-01-01

    Binding equilibria for decanoate to a defatted, commercially available human serum albumin preparation were investigated by dialysis exchange rate determinations. The binding isotherm could not be fitted by the general binding equation. It was necessary to assume that the preparation was a mixture...... of two albumin components about 40% of the albumin having high affinity and about 60% having low affinity. By affinity chromatography we succeeded in purifying the low-affinity component from the mixture. The high-affinity component, however, could not be isolated. We further analyzed the fatty acid...... and drug binding abilities of the low-affinity component. The fatty acids decanoate, laurate, myristate and palmitate were bound with higher affinity to the mixture than to the low-affinity component. Diazepam was bound with nearly the same affinity to the low-affinity component as to the albumin mixture...

  7. Improvement of a measurement method of purified flows in a reflector of HANARO by an ultra-sonic flowmeter

    International Nuclear Information System (INIS)

    Choi, Young-San; Bae, Sang-Hoon; Kang, In-Hyuk; Lee, Yong-Sub; Jung, Hoan-Sung

    2007-01-01

    Heavy water is used in the reflector system in HANARO and the flow in the system is measured by a flowmeter and indicated in a control room. The Turbine Flowmeter to measure the purified flow, which had been used from the start up of reactor was broken down in the end of 2001. In order to avoid the exposure of tritium generated from heavy water leaked during a replacement, instead of fixing the flowmeter, an ultrasonic flowmeter was selected and installed and has been used to measure the flow. This paper describes the measurement principles, issues and calibration errors of the turbine flowmeter that was broken down. Also, it explains in detail the measurement principles of the ultrasonic flowmeter, the results of its field test and the results of its periodic tests for five years after the installation

  8. Bacterial Reaction Centers Purified with Styrene Maleic Acid Copolymer Retain Native Membrane Functional Properties and Display Enhanced Stability**

    Science.gov (United States)

    Swainsbury, David J K; Scheidelaar, Stefan; van Grondelle, Rienk; Killian, J Antoinette; Jones, Michael R

    2014-01-01

    Integral membrane proteins often present daunting challenges for biophysical characterization, a fundamental issue being how to select a surfactant that will optimally preserve the individual structure and functional properties of a given membrane protein. Bacterial reaction centers offer a rare opportunity to compare the properties of an integral membrane protein in different artificial lipid/surfactant environments with those in the native bilayer. Here, we demonstrate that reaction centers purified using a styrene maleic acid copolymer remain associated with a complement of native lipids and do not display the modified functional properties that typically result from detergent solubilization. Direct comparisons show that reaction centers are more stable in this copolymer/lipid environment than in a detergent micelle or even in the native membrane, suggesting a promising new route to exploitation of such photovoltaic integral membrane proteins in device applications. PMID:25212490

  9. High level of microsynteny and purifying selection affect the evolution of WRKY family in Gramineae.

    Science.gov (United States)

    Jin, Jing; Kong, Jingjing; Qiu, Jianle; Zhu, Huasheng; Peng, Yuancheng; Jiang, Haiyang

    2016-01-01

    The WRKY gene family, which encodes proteins in the regulation processes of diverse developmental stages, is one of the largest families of transcription factors in higher plants. In this study, by searching for interspecies gene colinearity (microsynteny) and dating the age distributions of duplicated genes, we found 35 chromosomal segments of subgroup I genes of WRKY family (WRKY I) in four Gramineae species (Brachypodium, rice, sorghum, and maize) formed eight orthologous groups. After a stepwise gene-by-gene reciprocal comparison of all the protein sequences in the WRKY I gene flanking areas, highly conserved regions of microsynteny were found in the four Gramineae species. Most gene pairs showed conserved orientation within syntenic genome regions. Furthermore, tandem duplication events played the leading role in gene expansion. Eventually, environmental selection pressure analysis indicated strong purifying selection for the WRKY I genes in Gramineae, which may have been followed by gene loss and rearrangement. The results presented in this study provide basic information of Gramineae WRKY I genes and form the foundation for future functional studies of these genes. High level of microsynteny in the four grass species provides further evidence that a large-scale genome duplication event predated speciation.

  10. Properties of purified cytosolic isoenzyme I of Cu,Zn-superoxide dismutase from Nicotiana plumbaginifolia leaves.

    Science.gov (United States)

    Ragusa, S; Cambria, M T; Scarpa, M; Di Paolo, M L; Falconi, M; Rigo, A; Cambria, A

    2001-11-01

    The isoenzyme I of cytosolic Cu,Zn-superoxide dismutase (SOD) from Nicotiana plumbaginifolia (tobacco) leaves has been purified to apparent homogeneity. The relative molecular mass of the native isoenzyme, determined by gel filtration chromatography, is about 33.2 kDa. SDS-polyacrylamide gel electrophoresis shows that the enzyme is composed of two equal subunits of 16.6 kDa The isolectric point, assayed by isoelectric focusing, in the pH range of 3.5-6.5, is 4.3. The enzyme stability was tested at different temperatures, pH, and concentration of inhibitors (KCN and H(2)O(2)). The catalytic constant (k(cat)) was 1.17 +/- 0.14 x 10(9) M(-1) s(-1) at pH 9.9 and 0.1 M ionic strength. The activation energy of the thermal denaturation process is 263 kJ mol(-1). The electrostatic surface potential of the modeled tobacco Cu,Zn-SOD I was calculated showing that the functional spatial network of charges on the protein surface has been maintained, independently of the amino acid substitution around the active sites. Copyright 2001 Academic Press.

  11. All-Optical dc Nanotesla Magnetometry Using Silicon Vacancy Fine Structure in Isotopically Purified Silicon Carbide

    Directory of Open Access Journals (Sweden)

    D. Simin

    2016-07-01

    Full Text Available We uncover the fine structure of a silicon vacancy in isotopically purified silicon carbide (4H-^{28}SiC and reveal not yet considered terms in the spin Hamiltonian, originated from the trigonal pyramidal symmetry of this spin-3/2 color center. These terms give rise to additional spin transitions, which would be otherwise forbidden, and lead to a level anticrossing in an external magnetic field. We observe a sharp variation of the photoluminescence intensity in the vicinity of this level anticrossing, which can be used for a purely all-optical sensing of the magnetic field. We achieve dc magnetic field sensitivity better than 100  nT/sqrt[Hz] within a volume of 3×10^{-7}mm^{3} at room temperature and demonstrate that this contactless method is robust at high temperatures up to at least 500 K. As our approach does not require application of radio-frequency fields, it is scalable to much larger volumes. For an optimized light-trapping waveguide of 3  mm^{3}, the projection noise limit is below 100  fT/sqrt[Hz].

  12. Optimization of Freeze Drying Conditions for Purified Pectinase from Mango (Mangifera indica cv. Chokanan Peel

    Directory of Open Access Journals (Sweden)

    Abdul Manap Mohd Yazid

    2012-03-01

    Full Text Available Response surface methodology (RSM along with central composite design (CCD was applied to optimize the freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan peel. The effect of pectinase content (−2.66, 62.66 mg/mL, Arabic gum (−1.21, 10.21%, w/v, and maltodextrin (0.73, 7.26%, w/v as independent variables on activity, yield, and storage stability of freeze-dried enzyme was evaluated. Storage stability of pectinase was investigated after one week at 4 °C and yield percentage of the enzyme after encapsulation was also determined. The independent variables had the most significant (p < 0.05 effect on pectinase activity and yield of the enzyme. It was observed that the interaction effect of Arabic gum and maltodextrin improved the enzymatic properties of freeze-dried pectinase. The optimal conditions for freeze-dried pectinase from mango peel were obtained using 30 mg/mL of pectinase content, 4.5 (%, w/v of Arabic gum, and 4 (%, w/v of maltodextrin. Under these conditions, the maximum activity (11.12 U/mL, yield (86.4% and storage stability (84.2% of encapsulated pectinase were achieved.

  13. Optimization of Freeze Drying Conditions for Purified Pectinase from Mango (Mangifera indica cv. Chokanan) Peel

    Science.gov (United States)

    Mehrnoush, Amid; Mustafa, Shuhaimi; Yazid, Abdul Manap Mohd

    2012-01-01

    Response surface methodology (RSM) along with central composite design (CCD) was applied to optimize the freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan) peel. The effect of pectinase content (−2.66, 62.66 mg/mL), Arabic gum (−1.21, 10.21%, w/v), and maltodextrin (0.73, 7.26%, w/v) as independent variables on activity, yield, and storage stability of freeze-dried enzyme was evaluated. Storage stability of pectinase was investigated after one week at 4 °C and yield percentage of the enzyme after encapsulation was also determined. The independent variables had the most significant (p < 0.05) effect on pectinase activity and yield of the enzyme. It was observed that the interaction effect of Arabic gum and maltodextrin improved the enzymatic properties of freeze-dried pectinase. The optimal conditions for freeze-dried pectinase from mango peel were obtained using 30 mg/mL of pectinase content, 4.5 (%, w/v) of Arabic gum, and 4 (%, w/v) of maltodextrin. Under these conditions, the maximum activity (11.12 U/mL), yield (86.4%) and storage stability (84.2%) of encapsulated pectinase were achieved. PMID:22489134

  14. α-Glucosidase inhibitory activities of fatty acids purified from the internal organ of sea cucumber Stichopus japonicas.

    Science.gov (United States)

    Nguyen, T H; Kim, S M

    2015-04-01

    α-Glucosidase inhibitory activities of the various solvent fractions (n-hexane, CHCl3 , EtOAc, BuOH, and water) of sea cucumber internal organ were investigated. 1,3-Dipalmitolein (1) and cis-9-octadecenoic acid (2) with potent α-glucosidase inhibitory activity were purified from the n-hexane fraction of sea cucumber internal organ. IC50 values of compounds 1 and 2 were 4.45 and 14.87 μM against Saccharomyces cerevisiae α-glucosidase. These compounds mildly inhibited rat-intestinal α-glucosidase. In addition, both compounds showed a mixed competitive inhibition against S. cerevisiae α-glucosidase and were very stable at pH 2 up to 60 min. The KI values of compounds 1 and 2 were 0.48 and 1.24 μM, respectively. Therefore, the internal organ of sea cucumber might be a potential new source of α-glucosidase inhibitors suitably used for prevention of obesity and diabetes mellitus. © 2015 Institute of Food Technologists®

  15. Structure characteristics of an acidic polysaccharide purified from banana (Musa nana Lour.) pulp and its enzymatic degradation.

    Science.gov (United States)

    Liu, Huiling; Jiang, Yueming; Yang, Hongshun; Yang, Bao

    2017-08-01

    Banana is one of the most important fruits over the world. The chemical composition is critical for the organoleptic properties and health benefits. As one of the leading bioactive components in banana pulp, the polysaccharides may contribute to the beneficial health effects. However, their precise structure information remains unknown. A leading acidic polysaccharide (ABPP) of banana pulp was purified and identified by nuclear magnetic resonnance spectroscopy (NMR) and gas chromatography-mass spectrometry (GC-MS). →4-α-d-GalpA-1→ and →4-α-d-GalpAMe-1→ constituted the backbone. No branch chains were detected. The molecular weight was determined to be 8.9kDa by gel permeation chromatography, which was smaller than previously reported fruit-derived polygalacturonic acids. The precise structure was identified as below. Digestion by enzyme would lead to production of oligogalacturonic acids and quick accumulation of 5000-7000Da fraction. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Commercial Milk Enzyme-Linked Immunosorbent Assay (ELISA) Kit Reactivities to Purified Milk Proteins and Milk-Derived Ingredients.

    Science.gov (United States)

    Ivens, Katherine O; Baumert, Joseph L; Taylor, Steve L

    2016-07-01

    Numerous commercial enzyme-linked immunosorbent assay (ELISA) kits exist to quantitatively detect bovine milk residues in foods. Milk contains many proteins that can serve as ELISA targets including caseins (α-, β-, or κ-casein) and whey proteins (α-lactalbumin or β-lactoglobulin). Nine commercially-available milk ELISA kits were selected to compare the specificity and sensitivity with 5 purified milk proteins and 3 milk-derived ingredients. All of the milk kits were capable of quantifying nonfat dry milk (NFDM), but did not necessarily detect all individual protein fractions. While milk-derived ingredients were detected by the kits, their quantitation may be inaccurate due to the use of different calibrators, reference materials, and antibodies in kit development. The establishment of a standard reference material for the calibration of milk ELISA kits is increasingly important. The appropriate selection and understanding of milk ELISA kits for food analysis is critical to accurate quantification of milk residues and informed risk management decisions. © 2016 Institute of Food Technologists®

  17. Measurement of IgE antibodies against purified grass pollen allergens (Lol p 1, 2, 3 and 5) during immunotherapy.

    Science.gov (United States)

    Van Ree, R; Van Leeuwen, W A; Dieges, P H; Van Wijk, R G; De Jong, N; Brewczyski, P Z; Kroon, A M; Schilte, P P; Tan, K Y; Simon-Licht, I F; Roberts, A M; Stapel, S O; Aalberse, R C

    1997-01-01

    IgE titres tend to rise early after the start of immunotherapy, followed by a decline to pre-immunotherapy levels or lower. We were interested to know whether the early increase in IgE antibodies includes new specificities of IgE, and whether these responses persist. Sera of 64 patients undergoing grass pollen immunotherapy were tested for IgE against four purified grass pollen allergens: Lol p 1, 2, 3, and 5. At least two serum samples were taken, one before the start of therapy and one between 5 and 18 months after the first immunization (mean: 10 months). The mean IgE responses to Lol p 1, 2 and 3 showed a moderate but not significant increase. In contrast, the mean IgE response to Lol p 5 showed a significant decrease of > 30%. IgE against total Lohum perenne pollen extract moderately increased (> 20%), showing that a RAST for total pollen is not always indicative for the development of IgE against its major allergens. For > 40% of the patients it was found that IgE against one or more of the four allergens increased, while IgE against the remaining allergen(s) decreased. For 10 sera the ratio of IgE titres against at least two allergens changed by at least a factor of 5. The changes in specific IgE also included conversions from negative (< 0.1 RU) to positive (0.6 to 5.0 RU) for five patients. For two patients, the induction of these 'new' IgE antibodies against major allergens was shown to result in a response that was persistent over several years. Although active induction of new IgE specificities by immunotherapy was not really proven, the observations in this study indicate that monitoring of IgE against purified (major) allergens is necessary to evaluate changes in specific IgE in a reliable way.

  18. Purifying Selection and Birth-and-Death Evolution in the Class II Hydrophobin Gene Families of the Ascomycete Trichoderma/Hypocrea

    Energy Technology Data Exchange (ETDEWEB)

    kubicek, Christian P.; Baker, Scott E.; Gamauf, Christian; Kenerley, Chuck; Druzhinina, Irina S.

    2008-01-10

    Hydrophobins are proteins containing eight conserved cysteine residues that occur uniquely in mycelial fungi, where their main function is to confer hydrophobicity to fungal surfaces in contact with air and during attachment of hyphae to hydrophobic surfaces of hosts, symbiotic partners or of themselves resulting in morphogenetic signals. Based on their hydropathy patterns and their solubility characteristics, they are classified in class I and class II hydrophobins, the latter being found only in ascomycetes. Here we have investigated the mechanisms driving the evolution of the class II hydrophobins in nine species of the mycoparasitic ascomycetous genus Trichoderma/Hypocrea, using three fully sequenced genomes (H. jecorina=T. reesei, H. atroviridis=T. atroviride; H. virens=T. virens) and a total of 14.000 ESTs of six others (T. asperellum, H. lixii=T. harzianum, T. aggressivum var. europeae, T. longibrachiatum, T. cf. viride). The former three contained six, ten and nine members, which is the highest number found in any other ascomycete so far. They all showed the conserved four beta-strands/one helix structure, which is stabilized by four disulfide bonds. In addition, a small number of these HFBs contained an extended N-terminus rich in either praline and aspartate, or glycine-asparagine. Phylogenetic analysis reveals a mosaic of terminal clades contain duplicated genes and shows only three reasonably supported clades. Calculation of the ratio of differences in synonymous vs. non-synonymous nucleotide substitutions provides evidence for strong purifying selection (KS/Ka >> 1). A genome database search for class II HFBs from other ascomycetes retrieved a much smaller number of hydrophobins (2-4) from each species, and most of them were from Pyrenomycetes. A combined phylogeny of these sequences with those of Trichoderma showed that the Trichoderma HFBs mostly formed their own clades, whereas those of other pyrenomycetes occured in shared clades. Our study shows

  19. Structure-guided design of an engineered streptavidin with reusability to purify streptavidin-binding peptide tagged proteins or biotinylated proteins.

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    Sau-Ching Wu

    Full Text Available Development of a high-affinity streptavidin-binding peptide (SBP tag allows the tagged recombinant proteins to be affinity purified using the streptavidin matrix without the need of biotinylation. The major limitation of this powerful technology is the requirement to use biotin to elute the SBP-tagged proteins from the streptavidin matrix. Tight biotin binding by streptavidin essentially allows the matrix to be used only once. To address this problem, differences in interactions of biotin and SBP with streptavidin were explored. Loop3-4 which serves as a mobile lid for the biotin binding pocket in streptavidin is in the closed state with biotin binding. In contrast, this loop is in the open state with SBP binding. Replacement of glycine-48 with a bulkier residue (threonine in this loop selectively reduces the biotin binding affinity (Kd from 4 × 10(-14 M to 4.45 × 10(-10 M without affecting the SBP binding affinity. Introduction of a second mutation (S27A to the first mutein (G48T results in the development of a novel engineered streptavidin SAVSBPM18 which could be recombinantly produced in the functional form from Bacillus subtilis via secretion. To form an intact binding pocket for tight binding of SBP, two diagonally oriented subunits in a tetrameric streptavidin are required. It is vital for SAVSBPM18 to be stably in the tetrameric state in solution. This was confirmed using an HPLC/Laser light scattering system. SAVSBPM18 retains high binding affinity to SBP but has reversible biotin binding capability. The SAVSBPM18 matrix can be applied to affinity purify SBP-tagged proteins or biotinylated molecules to homogeneity with high recovery in a reusable manner. A mild washing step is sufficient to regenerate the matrix which can be reused for multiple rounds. Other applications including development of automated protein purification systems, lab-on-a-chip micro-devices, reusable biosensors, bioreactors and microarrays, and strippable

  20. Structure-guided design of an engineered streptavidin with reusability to purify streptavidin-binding peptide tagged proteins or biotinylated proteins.

    Science.gov (United States)

    Wu, Sau-Ching; Wong, Sui-Lam

    2013-01-01

    Development of a high-affinity streptavidin-binding peptide (SBP) tag allows the tagged recombinant proteins to be affinity purified using the streptavidin matrix without the need of biotinylation. The major limitation of this powerful technology is the requirement to use biotin to elute the SBP-tagged proteins from the streptavidin matrix. Tight biotin binding by streptavidin essentially allows the matrix to be used only once. To address this problem, differences in interactions of biotin and SBP with streptavidin were explored. Loop3-4 which serves as a mobile lid for the biotin binding pocket in streptavidin is in the closed state with biotin binding. In contrast, this loop is in the open state with SBP binding. Replacement of glycine-48 with a bulkier residue (threonine) in this loop selectively reduces the biotin binding affinity (Kd) from 4 × 10(-14) M to 4.45 × 10(-10) M without affecting the SBP binding affinity. Introduction of a second mutation (S27A) to the first mutein (G48T) results in the development of a novel engineered streptavidin SAVSBPM18 which could be recombinantly produced in the functional form from Bacillus subtilis via secretion. To form an intact binding pocket for tight binding of SBP, two diagonally oriented subunits in a tetrameric streptavidin are required. It is vital for SAVSBPM18 to be stably in the tetrameric state in solution. This was confirmed using an HPLC/Laser light scattering system. SAVSBPM18 retains high binding affinity to SBP but has reversible biotin binding capability. The SAVSBPM18 matrix can be applied to affinity purify SBP-tagged proteins or biotinylated molecules to homogeneity with high recovery in a reusable manner. A mild washing step is sufficient to regenerate the matrix which can be reused for multiple rounds. Other applications including development of automated protein purification systems, lab-on-a-chip micro-devices, reusable biosensors, bioreactors and microarrays, and strippable detection agents for