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Sample records for maculatus fab coleoptera

  1. Activités insecticides de Striga hermonthica (Del. Benth (Scrophulariaceae sur Callosobrichus maculatus (Fab. (Coleoptera : Bruchidae

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    Nacoulma OG.

    2006-01-01

    Full Text Available Insecticidal activities of Striga hermonthica (Del. Benth (Scrophulariacecae on Callobruchus maculatus (Fab. (Coleptera Bruchidae. This paper deals with insecticidal potentialities of Striga hermonthica (Del. (Scrophulariaceae in protection of cowpea Vigna unguculata (L. Walp against Callosobruchus maculatus (Fab. (Coleoptera: Bruchidae during storage. Crude acetone extract at 0,5% w/w (100 mg of extract for 20 g of grain exhibits 48% of ovicidal effect and then reduces by half emergence rate of adult beetles at the first generation. This extract shows a weak insecticide activity against adults of C. maculatus. Petroleum ether fraction (0,4% w/w of the crude extract reveals ovicidal (51% and larvicidal (72% effects which reduce the emergence rate of adults to only 9%. LD50 and LD90 are monitored during crude extract fractionation to follow ovicidal and larvicidal compounds and to evaluate their efficacy during the isolation procedure. One fraction, mainly composed of two triterpenoid compounds has been identified as responsible of the ovicidal activity of S. hermonthica while the origin of the larvicidal activity hasn’t been identified.

  2. Observations on the Biology of Callosobruchus maculatus (Fab ...

    African Journals Online (AJOL)

    Observations on the biology of Callosobruchus maculatus( Fab.) were carried out under ambient laboratory conditions. Pattern of oviposition, female adult longevity and F1 progeny emergence in Callosobruchus maculatus ( Fab. ) were observed. The results showed that over 60% of the total number of eggs was laid during ...

  3. Attraction of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) to four varieties of Lathyrus sativus L. seed volatiles.

    Science.gov (United States)

    Adhikary, P; Mukherjee, A; Barik, A

    2015-04-01

    Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) is an important stored grain pest of Lathyrus sativus L. (Leguminosae), commonly known as khesari, in India, Bangladesh and Ethiopia. Volatiles were collected from four varieties, i.e., Bio L 212 Ratan, Nirmal B-1, WBK-14-7 and WBK-13-1 of uninfested khesari seeds, and subsequently identified and quantified by gas chromatography mass spectrometry and gas chromatography flame ionization detector analyses, respectively. A total of 23 volatiles were identified in the four varieties of khesari seeds. In Bio L 212 Ratan and WBK-13-1 seeds, nonanal was the most abundant followed by farnesyl acetone; whereas farnesyl acetone was predominant followed by nonanal in Nirmal B-1 and WBK-14-7 khesari seeds. The olfactory responses of female C. maculatus toward volatile blends from four varieties of khesari seeds, and individual synthetic compounds and their combinations were examined through Y-shaped glass tube olfactometer bioassays. Callosobruchus maculatus showed significant preference for the whole volatile blends from Bio L 212 Ratan seeds compared to whole volatile blends from other three varieties. The insect exhibited attraction to five individual synthetic compounds, 3-octanone, 3-octanol, linalool oxide, 1-octanol and nonanal. A synthetic blend of 448, 390, 1182, 659 and 8114 ng/20 μl methylene chloride of 3-octanone, 3-octanol, linalool oxide, 1-octanol and nonanal, respectively, was most attractive to C. maculatus, and this combination might be used for insect pest management program such as baited traps.

  4. Gamma radiation effects on Callosobruchus Maculatus (Fabr. , 1792) (Coleoptera, Bruchidae)

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    Walder, J M.M.

    1974-01-01

    The effect of gamma irradiation from a Cobalt-60 source on Callosobruchus maculatus (Fabr.) was studied. Experiments showed that at a dose-rate of 9820 +- 300 rad/hour the LD/sub 100/ of egg embryos was 2,2 Krad of larvae 3 Krad of pupae 22,8 Krad. Total sterilization of adults irradiated of the pupal stage was 5 Krad. The sterilizing dose of irradiated adults was 8 krad at a calculated dose-rate of 57.630+-5000 rad/hour, whereas the LD/sub 100/ was 330 krad at a dose-rate of 63650+-1500 rad/hour. It was further verified that the number of eggs laid decrease in the females and diminish in the males as a result of irradiation.

  5. ATIVIDADE INSETICIDA DE PLANTAS MEDICINAIS SOBRE O Callosobruchus maculatus (COLEOPTERA: BRUCHIDAE

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    Antonia Mirian Nogueira de Moura Guerra

    2009-01-01

    Full Text Available This study aimed to evaluate the insecticidal activity of eight medicinal plants on Callosobruchus maculatus (F. (Coleoptera: Bruchidae. Plant powder from Rosmarinus officinalis (L. leaves, Peumus boldus (Mol leaves, Matricaria chamomilla (L. flowers, Baccharis trimera (Less. leaves, Camellia sinensis (L. leaves, Thea sinensis (L. leaves, Ilex paraguariensis (St. Hil. leaves, and fruits of Pimpinella anisum (L. were used in the experiment. Bioassays were carried out under constant conditions of temperature (28±2 °C, relative humidity (70±5% and scotophase of 24 hours. Experimental units consisted of 20 grams of cowpea (Vigna unguiculata (L. Walp. grains with 13% wet basis moisture content, 2 g plant powder and ten adult insects, both placed in plastic containers (60 mm in diameter x 60 mm high. The analyzed variables included: insect mortality, oviposition (egg number and instantaneous rate of population growth (ri. The number of dead insects was recorded 5 days after bioassays were started, the egg number was recorded after 12 days, and ri was estimated on the 60th day. All the tested plants had toxicity to C. maculatus. The highest mortality was found in insects treated with P. boldus and I. paraguariensis. Number of eggs and ri also varied significantly among the powders. The lowest mean values for these parameters were recorded in the plots treated with P. boldus, P. anisum and R. officinalis.

  6. Some effects of gamma radiation in Callosobruchus maculatus (Fabr., 1792) (Coleoptera, Bruchidae); Alguns efeitos da radiacao gama em Callosobruchus maculatus (Fabr., 1792) (Coleoptera, Bruchidae)

    Energy Technology Data Exchange (ETDEWEB)

    Walder, J M.M.

    1975-12-31

    The effect of gamma irradiation from a Cobalt-60 source on Callosobruchus maculatus (Fabr.) was studied in this work. Experiments showed that at a dose-rate of 9820 {+-} 300 rad/hour the LD sub(100) of egg embryos was 2,2 krad, of larvae 3 krad and of pupae 22,8 krad. Total sterilization of adults irradiated of the pupal stage was 5 krad. The sterilizing dose of irradiated adults was 8 krad at a calculated dose-rate of 57.630 {+-} 5000 rad/hour, whereas the LD sub(100) was 330 krad at a dose-rate of 63650 {+-} 1500 rad/hour. It was further verified that the number of eggs laid decreased with the irradiation dose. Longevity tended by increase in the females and diminish in the males as a result of irradiation. (author).

  7. Alternation of cowpea genotypes affects the biology of Callosobruchus maculatus (fabr. (Coleoptera: Bruchidae

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    Lima Marcileyne Pessôa Leite de

    2004-01-01

    Full Text Available Callosobruchus maculatus (Fabr. is an important pest in stored cowpea, Vigna unguiculata (L. Walp., with ample distribution in tropical and subtropical regions. The effect of alternation of cowpea genotypes, susceptible (S and resistant (R, on the biology of (C. maculatus was studied after four generations. A no-choice test was carried out in a completely randomized design, factorial scheme, with five treatments, four host combinations (RR, RS, SR and SS and five replications. Each replication consisted of 30 grains of each genotype infested by two insect couples. The number of eggs per female was not different within or between combinations, evidencing that the genotypes and their alternation did not affect C. maculatus fecundity. Egg viability, however, varied between genotypes and between combinations. In combination RR, the longest duration of the immature stage was verified for genotype IT89KD-245; in addition, all genotypes presented the smallest survival for the same stage, resulting in a higher mortality of the pest. The resistance index categorized combination RR as moderately resistant for genotypes IT89KD-245, BR14-Mulato and BR17-Gurguéia, and as susceptible (S only for IT89KD-260, demonstrating that these combinations were not very adequate for the development of C. maculatus, a fact that was confirmed by the better performance of the pest on the genotype from combination SS, and because of a reduction in its performance when it returned to resistant genotypes.

  8. Control of Cowpea Weevil, Callosobruchus Maculatus (F.) (Coleoptera: Bruchidae), Using Natural Plant Products

    OpenAIRE

    Tiroesele, Bamphitlhi; Thomas, Kesegofetse; Seketeme, Seipati

    2014-01-01

    A laboratory study was conducted to investigate the effects of natural products on the reproduction and damage of Callosobruchus maculatus, the cowpea weevil, on cowpea seeds at Botswana College of Agriculture in Gaborone, Botswana. The cowpea variety Blackeye was used in the study. Fifty grams of each plant product (garlic, peppermint and chilies) was added to 500 g of the cowpea seeds. Findings of this experiment revealed that chilies and garlic had negative effects on cowpea weevils for al...

  9. Gamma radiation influence on oviposition and longevity of 'Callosobruchus maculatus' (Fabr., 1972) (Coleoptera, Bruchidae)

    International Nuclear Information System (INIS)

    Walder, J.M.M.; Wiendl, F.M.

    1973-01-01

    It was verified that 'Callosobruchus maculatus' (Fabr.), a serious pest attacking seeds of 'Vigna sinensis', is highly susceptible to gamma radiation for sterilization. A dosis of 5 krad causes the egg fertility to decrease from 90% to only 1%. Adults irradiated with 10 krad become totally infertile. Doses of 10 krad and more cause an increase of about 19% in the longevity of females, but decreases the longetivy of males by about 5%. Also, irradiation causes a decrease in the total number of eggs, inversely proportional to the dosage increase [pt

  10. Gamma radiation influence on oviposition and longevity of 'Callosobruchus maculatus' (Fabr. , 1972) (Coleoptera, Bruchidae)

    Energy Technology Data Exchange (ETDEWEB)

    Walder, J M.M. [Centro de Energia Nuclear na Agricultura, Piracicaba (Brazil); Wiendl, F M [Sao Paulo Univ., Piracicaba (Brazil). Escola Superior de Agricultura Luiz de Queiroz

    1973-03-01

    It was verified that 'Callosobruchus maculatus' (Fabr.), a serious pest attacking seeds of 'Vigna sinensis', is highly susceptible to gamma radiation for sterilization. A dosis of 5 krad causes the egg fertility to decrease from 90% to only 1%. Adults irradiated with 10 krad become totally infertile. Doses of 10 krad and more cause an increase of about 19% in the longevity of females, but decreases the longevity of males by about 5%. Also, irradiation causes a decrease in the total number of eggs, inversely proportional to the dosage increase.

  11. Oxygen Consumption and Acoustic Activity of Adult Callosobruchus maculatus (F.) (Coleoptera: Chrysomelidae: Bruchinae) during Hermetic Storage.

    Science.gov (United States)

    Njoroge, Anastasia W; Mankin, Richard W; Smith, Bradley W; Baributsa, Dieudonne

    2018-04-20

    Acoustic monitoring was applied to consider hermetic exposure durations and oxygen levels required to stop adult Callosobruchus maculatus activity and economic damage on cowpea. A 15-d study was conducted with six treatments of 25, 50, and 100 C. maculatus adults in 500 and 1000 mL jars using acoustic probes inserted through stoppers sealing the jars. Acoustic activity as a result of locomotion, mating, and egg-laying was measured by identifying sound impulses with frequency spectra representative of known insect sounds, and counting trains (bursts) of impulses separated by intervals of <200 ms, that typically are produced only by insects. By the end of the first week of storage in all treatments, oxygen levels declined to levels below 4%, which has been demonstrated to cause mortality in previous studies. Concomitantly, insect sound burst rates dropped below an acoustic detection threshold of 0.02 bursts s −1 , indicating that the insects had ceased feeding. Statistically significant relationships were obtained between two different measures of the acoustic activity and the residual oxygen level. Based on the experimental results, a simple equation can be used to estimate the time needed for oxygen to decline to levels that limit insect feeding damage and thus grain losses in hermetic storage containers of different insect population levels and various volumes.

  12. Condition dependence of male and female genital structures in the seed beetle Callosobruchus maculatus (Coleoptera: Bruchidae).

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    Cayetano, L; Bonduriansky, R

    2015-07-01

    Theory predicts that costly secondary sexual traits will evolve heightened condition dependence, and many studies have reported strong condition dependence of signal and weapon traits in a variety of species. However, although genital structures often play key roles in intersexual interactions and appear to be subject to sexual or sexually antagonistic selection, few studies have examined the condition dependence of genital structures, especially in both sexes simultaneously. We investigated the responses of male and female genital structures to manipulation of larval diet quality (new versus once-used mung beans) in the bruchid seed beetle Callosobruchus maculatus. We quantified effects on mean relative size and static allometry of the male aedeagus, aedeagal spines, flap and paramere and the female reproductive tract and bursal spines. None of the male traits showed a significant effect of diet quality. In females, we found that longer bursal spines (relative to body size) were expressed on low-quality diet. Although the function of bursal spines is poorly understood, we suggest that greater bursal spine length in low-condition females may represent a sexually antagonistic adaptation. Overall, we found no evidence that genital traits in C. maculatus are expressed to a greater extent when nutrients are more abundant. This suggests that, even though some genital traits appear to function as secondary sexual traits, genital traits do not exhibit heightened condition dependence in this species. We discuss possible reasons for this finding. © 2015 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2015 European Society For Evolutionary Biology.

  13. Effect of gamma radiation on the metamorphic stages of Dermestes maculatus DeGeer (Coleoptera: Dermestidae)

    International Nuclear Information System (INIS)

    Seal, D.R.; Tilton, E.W.

    1986-01-01

    The effects of γ-radiation on all stages of the hide beetle Dermestes maculatus, DeGeer were studied. Eggs of D. maculatus were more susceptible to γ radiation than other stages. Egg radiosensitivity decreased with increasing embryonic development. An absorbed dose of 200 Gy killed the 1st, 6th and 7th instar larvae, but the 4th and 5th instar larvae were more resistant. The developmental period increased in treated larvae. Pupae (24 h) treated with 150 Gy failed to eclose, but eclosion was not affected in older pupae. Adults from female pupae irradiated at 72 h with 150 Gy were infertile, but male pupae required more than 200 Gy for sterilization. The average number of eggs per female decreased with increasing doses when either the male or female of the pair was irradiated as pupae or adults. Adult males were sterile after irradiation 300 Gy and adult females treated with the same dose failed to lay eggs. Newly emerged irradiated adults or female adults from irradiated 72-h-old pupae recovered some fertility after treatment with doses as high as 150 Gy. Adult males from irradiated 72-h-old pupae were treated at doses of 50 and 100 Gy showed a higher reproductive capacity at 60 days than at 15 days post-treatment. A dose between 200 and 300 Gy was necessary to provide complete sterility of 24-h-old adults. (author)

  14. Insecticidal activity of 2-tridecanone against the cowpea weevil Callosobruchus maculatus (Coleoptera: Bruchidae

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    Yussef F.B. Braga

    2007-03-01

    Full Text Available The effect of 2-tridecanone vapor on the cowpea weevil (Callosobruchus maculatus development was determined. Seeds of cowpea were infested with adults and exposed to different doses of 2-tridecanone isolated from Pilocarpus microphyllus Stapf ex Holm, a plant species native from northeastern Brazil. The pure monoterpene was evaluated both undiluted as well as in the dilutions 1:10, 1:100 and 1:1,000 (v/v. The following parameters of the cowpea weevil life cycle were analyzed in response to decreasing doses of 2-tridecanone: number of eggs laid, percentage of egg hatching on seeds, percentage of adult emergence, adult weight at emergence, mean developmental time and number of adults emerged. Vapor of 2-tridecanone caused a significant (P O efeito dos vapores da 2-tridecanona sobre o caruncho do feijão-de-corda (Callosobruchus maculatus foi avaliado. Sementes de feijão-de-corda infestados com insetos adultos foram expostas a diferentes doses de 2-tridecanona isolada de Pilocarpus microphyllus, uma espécie nativa do Nordeste do Brasil. O monoterpeno puro foi utilizado nas diluições 1:10, 1:100 e 1:1000 (v/v. Os parâmetros da biologia do inseto foram analisados em função da resposta a doses decrescentes de 2-tridecanona: número de ovos postos por fêmea, percentagem de eclosão de ovos, percentagem de emergência de adultos, peso dos adultos recém-emergidos, tempo médio de desenvolvimento e número total de ovos emergidos. Diferenças significativas (P < 0.05 entre as doses de 2-tridecanona testadas foram observadas, para quatro dos seis parâmetros biológicos analisados. Os resultados obtidos indicaram que a 2-tridecanona é tóxica para C. maculatus, reduzindo significativamente (P < 0.05 o número de insetos emergidos após a infestação. Esse efeito foi causado principalmente pela significativa redução observada na eclosão dos ovos expostos ao vapor da substância.

  15. Irradiation disinfestation of cowpea seeds to control southern cowpea beetle Callosbruchus maculatus (Fabricius) (Coleoptera Bruchidae)

    International Nuclear Information System (INIS)

    El-Kady, E.A.; Hekal, A.M.; El-Refai, S.

    1987-01-01

    The possibility of using gamma radiation as an alternative to fumigation or direct chemical applications for the control of stored-products insects was investigated in the laboratory and in a warehouse in Cairo, Egypt. Polyethylene bags of the type used in local markets were filled with cowpeas [Vigna unguiculata] Callosobruchus maculatus and irradiated at 400, 600 or 800 Gy [4, 6 or 8 krad]. All the treatments were effective, and no living adults infested by were found in the bags until 3 months later, when adults from outside penetrated the bags with their mandibles. It is therefore recommended that 4 krad should be administered for initial control, but that treated cowpeas should be stored in polypropylene rather than polyethylene bags in order to prevent reinfestation

  16. Global proteome changes in larvae of Callosobruchus maculatus Coleoptera:Chrysomelidae:Bruchinae) following ingestion of a cysteine proteinase inhibitor

    DEFF Research Database (Denmark)

    Nogueira, Fábio C S; Silva, Carlos P; Alexandre, Daniel

    2012-01-01

    The seed-feeding beetle Callosobruchus maculatus is an important cowpea pest (Vigna unguiculata) as well as an interesting model to study insect digestive physiology. The larvae of C. maculatus rely on cysteine and aspartic peptidases to digest proteins in their diet. In this work, the global...

  17. Fumigant Toxicity of Crushed Bulbs of Two Allium Species to Callosobruchus maculatus (Fabricius (Coleoptera: Bruchidae Toxicidad Fumigante para Callosobruchus maculatus (Fabriciu (Coleoptera: Bruchidae de Bulbos Trozados de Especies Allium

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    T.I Ofuya

    2010-09-01

    Full Text Available Fumigant toxicity of crushed fresh bulbs of Allium sativum L. and A. cepa L. to the Callosobruchus maculatus (Fabricius, a major pest of stored cowpea (Vigna unguiculata (L. Walp. seeds was assessed under laboratory conditions in Akure, Nigeria. In the tests, 20 g of infested cowpea seeds were suspended in a piece of muslin cloth, over an amount of crushed bulb in a container with a tightly fitted lid. Adult emergence was completely prevented from freshly laid eggs of C. maculatus on cowpea seeds that was fumigated with 6.0 g or more of crushed bulbs of A. sativum. Such fumigated seeds were not holed at all. Other amounts of A. sativum tested (1.0, 2.0, 3.0, 4.0 and 5.0 g significantly reduced C. maculatus adult emergence from fumigated eggs and seed holing in comparison with the control. Crushed A. sativum was ineffective in preventing adult emergence from fumigated C. maculatus larvae in seeds. The fumigant effect of crushed A. cepa did not kill all C. maculatus eggs. An amount of 7.0 g significantly reduced C. maculatus adult emergence from fumigated eggs and seed holing in comparison with the control. There is good prospect in using crushed bulbs of A. sativum> as fumigant in C. maculatus control in stored cowpea seeds.Se evaluó la toxicidad fumigante de bulbos frescos trozados de Allium sativum L. y A. cepa L. sobre Callosobruchus maculatus (Fabricius, una importante plaga de semilla almacenada de caupí Vigna unguiculata (L. Walp. bajo condiciones de laboratorio en Akure, Nigeria. En las pruebas, 20 g de semillas infestadas se suspendieron en un trozo de tela sobre cierta cantidad de bulbos trozados en un contenedor con una tapa ajustada. Se previno completamente la emergencia de adultos desde huevos recién puestos de C. maculatus en semillas de caupí que se fumigaron con 6,0 g o más de bulbos de A. sativum. Estas semillas fumigadas no estaban ahuecadas. Otras cantidades de A. sativum probadas (1.0, 2.0, 3.0, 4.0 y 5.0 g redujeron

  18. Bioactivity of Powder and Extracts from Garlic, Allium sativum L. (Alliaceae and Spring Onion, Allium fistulosum L. (Alliaceae against Callosobruchus maculatus F. (Coleoptera: Bruchidae on Cowpea, Vigna unguiculata (L. Walp (Leguminosae Seeds

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    Abiodun A. Denloye

    2010-01-01

    Full Text Available Laboratory bioassays were conducted to investigate the bioactivity of powders, extracts, and essential oils from Allium sativum L. (Alliaceae and A. fistulosum L. (Liliaceae against adults, eggs, and larvae of Callosobruchus maculatus F. (Coleoptera: Bruchidae. On the basis of 48 hr median lethal toxicity (LC50, test plant powders and extracts from A. sativum were more toxic to C. maculatus adults than those from A. fistulosum. The 48 hr LC50 values for the powder against the test insect species were 9.66 g/kg and 26.29 g/kg for A. sativum and A. fistulosum, respectively. Also the 48 hr LC50 values obtained show that aqueous extracts of the test plant species, 0.11 g/L (A. sativum and 0.411 g/L (A. fistulosum were more toxic to C. maculatus than the corresponding ethanol extracts. There was no significant difference in the toxicity of vapours from the two test plant species against C. maculatus, although A. sativum gave lower values. The study shows that A. sativum and A. fistulosum have potentials for protecting stored cowpea from damage by C. maculatus.

  19. The seed coat of Phaseolus vulgaris interferes with the development of the cowpea weevil [Callosobruchus maculatus (F. (Coleoptera: Bruchidae

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    Silva Luciana B.

    2004-01-01

    Full Text Available We have confirmed here that the seeds of the common bean (Phaseolus vulgaris, L. do not support development of the bruchid Callosobruchus maculatus (F., a pest of cowpea [Vigna unguiculata (L. Walp] seeds. Analysis of the testa (seed coat of the bean suggested that neither thickness nor the levels of compounds such as tannic acid, tannins, or HCN are important for the resistance. On the other hand, we have found that phaseolin (vicilin-like 7S storage globulin, detected in the testa by Western blotting and N-terminal amino acid sequencing, is detrimental to the development of C. maculatus. As for the case of other previously studied legume seeds (Canavalia ensiformis and Phaseolus lunatus we suggest that the presence of vicilin-like proteins in the testa of P. vulgaris may have had a significant role in the evolutionary adaptation of bruchids to the seeds of leguminous plants.

  20. Ovipositional Deterrence of Methanolic and Etherial Extracts of Five Plants to the Cowpea Bruchid, Callosobruchus maculatus (F. (Coleoptera: Bruchidae

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    E.A. Elhag

    1999-06-01

    Full Text Available Methanol and diethyl ether extracts of Harmal, Rhazya stricta Decne.; neem seed kernels, Azadirachta indica A.Juss; cloves, Syzygeum aromarticum (L.; citrus peel and Ramram, Heliotropium bacciferum (Forssk- were evaluated for their deterrence to oviposition by Callosobruchus maculatus (F. on chickpeas in choice tests. Both extracts of all materials significantly reduced oviposition on treated seeds. Maximum deterrent effects (91.8% were obtained in the neem seed methanol extract at 0.5% concentration, citrus peel O. l% ether extract (90.9%, R stricta 0.5% methanol extract (83.9%, and clove 0. 1% ether extract (80.0%. Methanol extracts of neem seeds and R. stricta evoked higher deterrent effects than their etherial extracts, whereas the responses for cloves and citrus peel were more pronounced in their ether extracts. H. bacirferum % deterrency due to both types of extracts were practically identical. The results encourage future incorporation of such plant extracts as ovipositional deterrents in stored-product lPM programmes.

  1. Electron beam irradiation: a technology for quarantine disinfestation of green gram seeds against Callsobruchus maculatus

    International Nuclear Information System (INIS)

    Bhalla, Shashi; Srinivasan, K.; Singh, Subadas; Thakur, Manju; Sharma, S.K.; Pramod, R.; Dwivedi, J.; Bapna, S.C.

    2010-01-01

    Green gram (Vigna radiata (L.) Wilkzec) an important legume crop in India is grown in 33.4 lakh hectares. India accounts for ∼ 60% of the world's green gram area but contributes only 47% of its production. Major constraint in storage is the pulse beetle Callosobruchus maculatus Fab. (Coleoptera: Bruchidae), which may cause damage upto 100%. It is widespread throughout tropics and sub-tropics with wide host range and also has different strains. Fumigation with methyl bromide (MB) has been the most widely applied management practice for its control. However, the ozone depleting effect of MB has led to restrictions in its use. Therefore, there is a need for an alternative strategy for controlling the pests. Irradiation, an approved technology by International Plant Protection Convention, seems to be a viable non-chemical, residue-free strategy. Therefore, present studies were conducted to see the efficacy of electron beam (EB) irradiation as quarantine disinfestation treatment against green gram seeds infested with different stages of the target pest, C. maculatus

  2. Resistência de genótipos de feijão-caupi [Vigna unguiculata (L. Walp.] ao Ataque do Caruncho Callosobruchus maculatus (Fabr. (Coleoptera: Chrysomelidae

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    Westerllanya Medeiros

    2017-04-01

    Abstract. The Cowpea weevil Callosobruchus maculatus (Fabr. is considered the main Prague during storage of grains of Cowpea [Vigna unguiculata (L. Walp.]. Due to the economic losses caused by this insect, is of paramount importance to develop studies to select varieties resistant to this pest. In this study assessed the effect of genotypes of V. unguiculata about the behavior and the development of C. maculatus in two consecutive generations. The study was conducted in laboratory with conditions of temperature and relative humidity monitored, in completely randomized design using BRS Epace 10, Capela, BRS Urubuquara, BRS Pajeu, Itaim, BRS Rouxinol, Pingo de ouro, BRS Corujinha, IT85 F-2687 and BR 17 Gurguéia genotypes with eight repetitions. In each genotype were confined 10 adults of c. maculatus for it was held the oviposition. After a period of five days, the adults were removed from the pots containing the genotypes. Waited-if the emergence of adults (first generation and new infestation was held in the same access to obtain the second generation. The parameters evaluated were number of eggs, egg viability, total number of adults emergency, viability of immature phase, average period of development and dry weight of adults. IT85 F2687 genotype presented resistance of non-preference for oviposition and the genotypes BR 17 Gurguéia and BRS Urubuquara presented antibiose type resistance in relation to C. maculatus. BR 17 Gurguéia genotype was the strongest among the genotypes studied in relation to c. maculatus. The genotypes Capela and Itaim were characterized as susceptible to C. maculatus.

  3. Biochemical studies of amylase, lipase and protease in Callosobruchus maculatus (Coleoptera: Chrysomelidae) populations fed with Vigna unguiculata grain cultivated with diazotrophic bacteria strains.

    Science.gov (United States)

    Silva, L B; Torres, É B; Nóbrega, R A S; Lopes, G N; Vogado, R F; Pavan, B E; Fernandes-Junior, P I

    2017-12-01

    The objective of this study was to evaluate the enzymatic activity of homogenates of insects fed on grain of cowpea, Vigna unguiculata (L.), cultivars grown with different nitrogen sources. For the experiment we used aliquots of the homogenate of 100 unsexed adult insects, emerged from 10 g of grain obtained from four cowpea cultivars: 'BRS Acauã', 'BRS Carijó', 'BRS Pujante', and 'BRS Tapaihum' grown under different regimes of nitrogen sources: mineral fertilizer, inoculation with strains of diazotrophs (BR 3267, BR 3262, BR 3299; INPA 03-11B, 03-84 UFLA, as well as the control (with soil nitrogen). The parameters evaluated were enzymatic activities of insect protease, amylase and lipase and the starch content of the grains. There were differences in the enzymatic activity of amylase, lipase and protease of insect homogenate according to the food source. A lower activity of the enzyme amylase from C. maculatus homogenate was observed when insects were fed grain of the cultivar BRS Carijó. A lower activity of lipase enzyme from C. maculatus homogenate was observed when the insects fed on grain from the interaction of the cultivar Tapaihum inoculated with BR 3262 diazotrophs. The lowest proteolytic activity was observed in homogenate of insects fed on interaction of 'BRS Carijó' inoculated with BR 3262 diazotrophs. Starch content correlated positively with the amylase activity of C. maculatus homogenate. The cultivar BRS Carijó had a different behavior from the other cultivars, according to the cluster analysis.

  4. Inherited influence of low dose gamma radiation on the reproductive potential and spermiogenesis of the cowpea weevil, Callosobruchus maculatus (F (Coleoptera: Chrysomelidae

    Directory of Open Access Journals (Sweden)

    Hatem A. Ibrahim

    2017-10-01

    Full Text Available The southern cowpea weevil, Callosobruchus maculatus (F is a severe agriculture pest worldwide. In the current work, newly emerged adult males of C. maculatus have been irradiated with a low dose of 20Gy gamma radiation. The inherited deleterious effects on the fecundity, hatchability, adult emergence, and the sterility percent were recorded for the progenies F1 and F2 of the irradiated parental males. The fecundity, hatched larvae, the number of males and females were reduced in both F1 and F2. The sterility percent was high in F1 (70.8% and increased in the F2 (88.3% generation. Histopathological effects were also documented in the testes of F1 and F2 progenies. The spermatids and sperms have exhibited a variety of abnormalities. In the early spermatids, the nebenkern outer cell membrane was ruptured. The spermatid nucleus loses its homogeneous texture and has multiple foci of dense chromatin, as well as, profiles range has little dense material. In some groups, the nucleus had a peculiar ring of chromatin. The sperms had shown a variety of aberrations. The sperms irregularity distributed in lysed cysts by unusual manner. Also, some sperms had remarkably enlarged axoneme and small rounded nucleus. Many of the sperm cells were observed with two axonemes, abnormal mitochondria derivatives, and more than two accessory bodies. These results indicate that the low dose of 20 Gy induces semi-sterility in C. maculatus through generations. The same technique would help to improve using of sterile insect technique for other agriculture pests.

  5. Evaluation of the seed oil of three citrus species, for the control of the bean beetle, callosobruchus maculatus (F) (coleoptera: bruchidae)

    International Nuclear Information System (INIS)

    Ogunleye, R.F.

    2009-01-01

    On application of the seed oil of ripe and unripe fruits of Citrus sinensis, C. paradisi and C. aurantifolia to the cowpea bruchid, Callosobruchus maculatus (F) for three days, a dose of 0.5 ml of C. sinensis gave, significantly, high mortality rate upto 85%. In case of C. aurantifolia, mortality ranged from 75% to 100%. Same least dosage of seed oil of ripe C. paradissi produced 58.8% to 100% mortality, whereas, except the dose of 0.5 ml, all the other treatments of unripe C. paradissi resulted in 100% mortality after 24 h. (author)

  6. Comparaison de quelques caractéristiques biologiques entre Dinarmus basalis Rond. (Hymenoptera: Pteromalidae élevé soit sur son hôte habituel Callosobruchus maculatus F. (Coleoptera: Bruchidae soit sur Acanthoscelides macrophthalmus Schaef. ou Bruchidius lineatopygus Pic. identifiés comme hôtes de substitution

    Directory of Open Access Journals (Sweden)

    Nuto, Y.

    2006-01-01

    Full Text Available Comparison of some Biological Characteristics between Dinarmus basalis Rond. (Hymenoptera: Pteromalidae Reared either on his Habitual Host Callosobruchus maculatus F. (Coleoptera: Bruchidae or on Acanthoscelides macrophthalmus Schaef. and Bruchidius lineatopygus Pic. Identified as Substitution Hosts. In this study, we have measured some biological parameters of Dinarmus basalis Rond. (Hymenoptera: Pteromalidae when this parasitoid grows on three hosts: Callosobruchus maculatus F. (usual host, Acanthoscelides macrophthalma Schaef. and Bruchidius lineatopygus Pic. (substitution hosts. The results of this comparative study in no choice situation show that, life duration and development duration of this parasitoid are identical in laboratory conditions. By contrast, the female fecundity, the sexual rate and the parasitism rate of D. basalis are different according to the hosts presented. Although such differences appear to be small between C. maculatus and A. macrophthalmus. In presence of these two species, the number of eggs laid by the female of D. basalis is respectively 61 ± 5.24 and 54 ± 5.60, the sexual rate 37.63 ± 2.13% and 41.73 ± 3.69% and the parasitism rate 85.46 ± 2.61% and 76.48 ± 5.90%. This indicates that these two Bruchids can be used as hosts to D. basalis for a mass production. In presence of B. lineatopygus, the fecundity and the parasitism rate of D. basalis female are very low and the sexual rate more favourable of the males. This situation is unfavourable for a parasitoid production. When D. basalis females are placed in choice situation with equal number of the hosts nowely C. maculatus and A. macrophthalmus, the parasitism rate obtained, is higher for C. maculatus that of A. macrophthalmus. This parasitism rate remains high for C. maculatus even when the other host A. macrophthalmus is twice the number of C. maculatus. Thus the parasitoid seems to have a preference for his natural host. However, A. macrophthalmus can

  7. Influência do período de armazenamento do caupi [Vigna unguiculata (L. Walp.], tratado com óleos essenciais e fixos, no controle de Callosobruchus maculatus (Fabricius, 1775 (Coleoptera, Chrysomelidae, Bruchinae Influence of the storage period of cowpea [Vigna unguiculata (L. Walp.] treated with essential and fixed oils, for the control of Callosobruchus maculatus (Fabricius, 1775 (Coleoptera, Chrysomelidae, Bruchinae

    Directory of Open Access Journals (Sweden)

    Adriana Carla Ribeiro Lopes Pereira

    2009-02-01

    Full Text Available Compostos secundários obtidos de plantas podem ser utilizados no controle de Callosobruchus maculatus, como uma tática alternativa potencial aos inseticidas sintéticos. Foram testados óleos essenciais (Cymbopogon martini Roxb., Piper aduncum L., Piper hispidinervum C.DC., Melaleuca sp., Lippia gracillis Shau e fixos (Helianthus annus L., Sesamum indicum L., Gossypium hirsutum L., Glycine max L. e Caryocar brasiliense Camb., na concentração de 50µl/20g, de acordo com estudos anteriores. Grãos de caupi, cv. Sempre Verde, foram impregnados com os óleos, em recipientes de vidro e submetidos à agitação manual por dois minutos. Cada parcela de 20g foi infestada com oito fêmeas de C. maculatus com 0 a 48h de idade, durante quatro dias. Os óleos foram avaliados logo após a impregnação e aos 30, 60, 90 e 120 dias de armazenamento. Na primeira avaliação, todos os óleos essenciais provocaram 100% de mortalidade e para os óleos fixos, a mortalidade variou entre 35% (G. hirsutum e 67,5% (G. max. Com o prolongamento do período de armazenamento, houve um aumento do número de ovos viáveis e de insetos emergidos, exceto para P. aduncum. Em relação aos óleos fixos, S. indicum, G. max, G. hirsutum e C. brasiliense foram os mais eficientes até os 30 dias de armazenamento. Os resultados indicam que os óleos testados na concentração de 50µl/20g apresentam baixo efeito residual, com exceção de P. aduncum, que foi efetivo durante todo o período de armazenamento.The secondary compounds extracted from plants are considered potential alternative to synthetic insecticides in the control of agricultural pests. Essential oils (Cymbopogon martini Roxb., Piper aduncum L., P. hispidinervum C.DC., Melaleuca sp. and Lippia gracillis Shau and fixed oils (Helianthus annus L., Sesamum indicum L., Gossypium hirsutum L., Glycine max L. and Caryocar brasiliense Camb. at the concentration of 50µl/20g were tested according to previous studies. Samples

  8. Atividade inseticida de óleos essenciais e fixos sobre Callosobruchus maculatus (FABR., 1775 (Coleoptera: Bruchidae em grãos de caupi [Vigna unguiculata (L. WALP.] Insecticide activity of essential and fixed oils in Callosobruchus maculatus (FABR., 1775 (Coleoptera: Bruchidae in cowpea grains [Vigna unguiculata (L. WALP.

    Directory of Open Access Journals (Sweden)

    Adriana Carla Ribeiro Lopes Pereira

    2008-06-01

    Full Text Available O caruncho, Callosobruchus maculatus (Fabr., é considerado a praga mais importante do caupi, Vigna unguiculata (L. Walp., armazenado em regiões tropicais e subtropicais. Visando minimizar os efeitos indesejáveis dos inseticidas químicos sintéticos, o controle dessa praga com óleos de origem vegetal vem se constituindo numa alternativa promissora, de baixo custo e segura para os aplicadores e consumidores. Foram testados os óleos essenciais [(Cymbopogon martini (Roxb. J.F. Watson], Piper aduncum L., Piper hispidinervum C.DC., Melaleuca sp., Lippia gracillis Shauer e fixos (Helianthus annuus L, Sesamum indicum L, Gossypium hirsutum L., Glycine max (L. Merr. e Caryocar brasiliense Camb., em grãos de caupi, cv. Sempre Verde. Os óleos foram utilizados nas concentrações 10, 20, 30, 40 e 50mL/20g, correspondendo a 0,5, 1,0, 1,5, 2,0 e 2,5 L/t e impregnados aos grãos no interior de recipientes de plástico, com auxílio de pipetador automático e agitados manualmente durante dois minutos. Parcelas de 20g de caupi foram infestadas com oito fêmeas de C. maculatus, com 0 a 48 h de idade. Cada óleo foi testado, separadamente, em delineamento inteiramente casualisado com seis repetições. Os óleos essenciais de C. martini, P. aduncum e L. gracillis causaram 100% de mortalidade em todas as concentrações, P. hispidinervum a partir de 1,5 L/t e Melaleuca sp. nas concentrações de 2,0 e 2,5 L/t. A redução do número de ovos viáveis e de insetos emergidos foi de 100% para todos os óleos essenciais, exceto Melaleuca sp. Por outro lado, os óleos fixos, apesar de apresentarem baixa mortalidade em todas as concentrações testadas, reduziram em praticamente 100% o número de ovos viáveis e de insetos emergidos.The weevil Callosobruchus maculatus (Fabr. is considered the most important pest of cowpea, Vigna unguiculata (L. Walp., stored in tropical and subtropical regions. Focusing to minimize the undesired effects of synthetic chemical

  9. Beyond Fab Four

    International Nuclear Information System (INIS)

    Babichev, E; Charmousis, C; Langlois, D; Saito, R

    2015-01-01

    We show that the two additional Lagrangians that appear in theories beyond Horndeski can be reexpressed in terms of simple generalizations of the ‘John’ and ‘Paul’ terms of the Fab Four theories. We find that these extended Fab Four satisfy the same properties of self-tuning as the original Fab Four. (fast track communication)

  10. FabIO

    DEFF Research Database (Denmark)

    Bergbäck Knudsen, Erik; Sørensen, Henning O.; Wright, Jonathan P.

    2013-01-01

    FabIO is a Python module written for easy and transparent reading of raw two-dimensional data from various X-ray detectors. The module provides a function for reading any image and returning a fabioimage object which contains both metadata (header information) and the raw data. All fabioimage...

  11. Fab Chaperone-Assisted RNA Crystallography (Fab CARC).

    Science.gov (United States)

    Sherman, Eileen; Archer, Jennifer; Ye, Jing-Dong

    2016-01-01

    Recent discovery of structured RNAs such as ribozymes and riboswitches shows that there is still much to learn about the structure and function of RNAs. Knowledge learned can be employed in both biochemical research and clinical applications. X-ray crystallography gives unparalleled atomic-level structural detail from which functional inferences can be deduced. However, the difficulty in obtaining high-quality crystals and their phasing information make it a very challenging task. RNA crystallography is particularly arduous due to several factors such as RNA's paucity of surface chemical diversity, lability, repetitive anionic backbone, and flexibility, all of which are counterproductive to crystal packing. Here we describe Fab chaperone assisted RNA crystallography (CARC), a systematic technique to increase RNA crystallography success by facilitating crystal packing as well as expediting phase determination through molecular replacement of conserved Fab domains. Major steps described in this chapter include selection of a synthetic Fab library displayed on M13 phage against a structured RNA crystallization target, ELISA for initial choice of binding Fabs, Fab expression followed by protein A affinity then cation exchange chromatography purification, final choice of Fab by binding specificity and affinity as determined by a dot blot assay, and lastly gel filtration purification of a large quantity of chosen Fabs for crystallization.

  12. "Fab 13": The Learning Factory.

    Science.gov (United States)

    Crooks, Steven M.; Eucker, Tom R.

    2001-01-01

    Describes how situated learning theory was employed in the design of Fab 13, a four-day simulation-based learning experience for manufacturing professionals at Intel Corporation. Presents a conceptual framework for understanding situated learning and discusses context, content, anchored instruction, facilitation, scaffolding, collaborating,…

  13. Efek Beauveria bassiana pada Anopheles maculatus Fase Aquatik di Laboratorium

    Directory of Open Access Journals (Sweden)

    Bina Ikawati

    2017-07-01

    Full Text Available AbstractBeauveria bassiana can be used both as for controlling agricultural insect and protecting health. Thisstudy aims to examine the effects of aquatic phase of B. bassiana on An. maculatus in the laboratory.Samples were eggs and larvae of An. maculatus reared from Banjarnegara colony. Eggs and instarlarvae II, III and IV and their control which consisted of 10 larvae/eggs were replicated six times andcontacted to B. bassiana spores for 15 minutes and then transferred to aquades to be maintained forobservation, in every 24 hours as long as 192 hours (8 days. Probit analysis found that applicationof B. bassiana caused damage of external coat of eggs and inhibited >60% of unhatched eggs. Lethaldosage was dosage spores of 1,713x107 (16 days, whereas the lethal dose required to make 50% ofunperached eggs is a dose of spore concentration of 1,361x107 (11.6 days. Higher concentrationswill be needed to know the faster effects B bassiana on An. maculatus larvae or eggs.Key words: Beauveria bassiana, Anopheles maculatus, aquatic phase, laboratory AbstrakBeauveria bassiana dapat digunakan baik sebagai pengendali serangga pertanian maupun kesehatan.Penelitian ini bertujuan mengkaji efek B. bassiana terhadap An. maculatus pada fase akuatik dilaboratorium. Sampel uji berupa telur dan larva An. maculatus dari koloni Banjarnegara. Telur danlarva instar II,III dan IV serta kontrol masing-masing sebanyak 10 ekor/butir dengan replikasi 6 kalidikontakkan dengan spora B. bassiana selama 15 menit dan selanjutnya dipindahkan ke aquadesuntuk dipelihara untuk dilakukan pengamatan, pengamatan dengan mikroskop compound terutamapada larva yang lemah/mati setiap 24 jam selama 192 jam (8 hari. Analisis probit membuktikanbahwa aplikasi B. bassiana pada telur menimbulkan efek kerusakan pada lapisan luar telur diketahuidari pengamatan dengan mikroskop serta mampu menghambat >60% telur tidak menetas. Simulasidengan análisis probit menunjukkan lethal dose yang dibutuhkan

  14. Nuclear DNA content in Galaxias maculatus (Teleostei: Osmeriformes: Galaxiidae Contenido de ADN nuclear en Galaxias maculatus (Teleostei: Osmeriformes: Galaxiidae

    Directory of Open Access Journals (Sweden)

    Pedro Jara-Seguel

    2008-01-01

    Full Text Available The nuclear DNA content (2C value was determined in the commercial fish Galaxias maculatus (Galaxiidae was determined by microdensitometry of erythrocyte nuclei after Feulgen staining; rainbow trout erythrocytes with a known 2C value were used as a standard. The 2C value of G. maculatus was 2.21 ± 0.12 pg and its C value was equivalent to 1.105 pg (1,082.9 Mbp. This C value is within the range recorded for other osmeriform species (0.62-3.2 pg. The average sperm head diameter of G. maculatus is lower than the average sperm head diameter of rainbow trout (used as a standard, which coincides with the differences observed in the nuclear DNA content of both species. This information increases the genome data available for G. maculatus and might be useful in future programs dealing with its genetic manipulation.El contenido de ADN nuclear (valor 2C fue determinado en el pez comercial Galaxias maculatus (Galaxiidae usando microdensitometría de núcleos de eritrocitos sometidos a tinción de Feulgen, utilizando como estándar eritrocitos de trucha arco iris con un valor 2C conocido. El valor 2C de G. maculatus fue 2,21 ± 0,12 pg y su valor C es equivalente a 1,105 pg (1.082,9 pMb. Este valor C está dentro del rango registrado para otras especies de osmeriformes (0,62-3,2 pg. El diámetro promedio de la cabeza del espermatozoide de G. maculatus es menor al promedio descrito para la trucha arco iris utilizado como estándar, lo que coincide con las diferencias observadas en el contenido de ADN nuclear entre ambas especies. Estos datos contribuyen a ampliar los antecedentes genómicos disponibles para G. maculatus y podrían ser útiles en futuros programas tendientes a su manipulación genética.

  15. Efficacy of plant extracts against the cowpea beetle, Callosobruchus maculatus

    NARCIS (Netherlands)

    Boeke, S.J.; Barnaud, B.; Loon, van J.J.A.; Kossou, D.K.; Huis, van A.; Dicke, M.

    2004-01-01

    Traditionally used African plant powders, with a known effect against the cowpea beetle Callosobruchus maculatus in stored cowpea, were extracted with water. The extracts, 13 volatile oils, 2 non-volatile oils and 8 slurries, were evaluated for their toxic and repellent effects against the beetle.

  16. Effect of Dermestes maculatus on the nutritional qualities of two ...

    African Journals Online (AJOL)

    This study aimed to assess the effect on nutritional composition of two 'Edible Insects' (Larva of Oryctes boas and Rhynchophorus phoenicis) after infestation by Dermestes maculatus. The study was done over a period of six weeks by subjecting these edible insects to different conditions of storage (using pepper and salt).

  17. Baculovirus display of functional antibody Fab fragments.

    Science.gov (United States)

    Takada, Shinya; Ogawa, Takafumi; Matsui, Kazusa; Suzuki, Tasuku; Katsuda, Tomohisa; Yamaji, Hideki

    2015-08-01

    The generation of a recombinant baculovirus that displays antibody Fab fragments on the surface was investigated. A recombinant baculovirus was engineered so that the heavy chain (Hc; Fd fragment) of a mouse Fab fragment was expressed as a fusion to the N-terminus of baculovirus gp64, while the light chain of the Fab fragment was simultaneously expressed as a secretory protein. Following infection of Sf9 insect cells with the recombinant baculovirus, the culture supernatant was analyzed by enzyme-linked immunosorbent assay using antigen-coated microplates and either an anti-mouse IgG or an anti-gp64 antibody. A relatively strong signal was obtained in each case, showing antigen-binding activity in the culture supernatant. In western blot analysis of the culture supernatant using the anti-gp64 antibody, specific protein bands were detected at an electrophoretic mobility that coincided with the molecular weight of the Hc-gp64 fusion protein as well as that of gp64. Flow cytometry using a fluorescein isothiocyanate-conjugated antibody specific to mouse IgG successfully detected the Fab fragments on the surface of the Sf9 cells. These results suggest that immunologically functional antibody Fab fragments can be displayed on the surface of baculovirus particles, and that a fluorescence-activated cell sorter with a fluorescence-labeled antigen can isolate baculoviruses displaying specific Fab fragments. This successful baculovirus display of antibody Fab fragments may offer a novel approach for the efficient selection of specific antibodies.

  18. Characterization of cowpea genotype resistance to Callosobruchus maculatus

    Directory of Open Access Journals (Sweden)

    Maria de Jesus Passos de Castro

    2013-09-01

    Full Text Available The objective of this work was to characterize the resistance of 50 cowpea (Vigna unguiculata genotypes to Callosobruchus maculatus. A completely randomized design with five replicates per treatment (genotype was used. No-choice tests were performed using the 50 cowpea genotypes to evaluate the preference for oviposition and the development of the weevil. The genotypes IT85 F-2687, MN05-841 B-49, MNC99-508-1, MNC99-510-8, TVu 1593, Canapuzinho-1-2, and Sanzi Sambili show non-preference-type resistance (oviposition and feeding. IT81 D-1045 Ereto and IT81 D-1045 Enramador exhibit antibiosis against C. maculatus and descend from resistant genitors, which grants them potential to be used in future crossings to obtain cowpea varieties with higher levels of resistance.

  19. Comparison of F(ab')2 versus Fab antivenom for pit viper envenomation: a prospective, blinded, multicenter, randomized clinical trial.

    Science.gov (United States)

    Bush, Sean P; Ruha, Anne-Michelle; Seifert, Steven A; Morgan, David L; Lewis, Brandon J; Arnold, Thomas C; Clark, Richard F; Meggs, William J; Toschlog, Eric A; Borron, Stephen W; Figge, Gary R; Sollee, Dawn R; Shirazi, Farshad M; Wolk, Robert; de Chazal, Ives; Quan, Dan; García-Ubbelohde, Walter; Alagón, Alejandro; Gerkin, Richard D; Boyer, Leslie V

    2015-01-01

    Crotalidae Polyvalent Immune Fab (Ovine) has been the only antivenom commercially available in the US since 2007 for treatment of Crotalinae envenomation. Late coagulopathy can occur or recur after clearance of Fab antivenom, often after hospital discharge, lasting in some cases more than 2 weeks. There have been serious, even fatal, bleeding complications associated with recurrence phenomena. Frequent follow-up is required, and additional intervention or hospitalization is often necessary. F(ab')2 immunoglobulin derivatives have longer plasma half life than do Fab. We hypothesized that F(ab')2 antivenom would be superior to Fab in the prevention of late coagulopathy following treatment of patients with Crotalinae envenomation. We conducted a prospective, double-blind, randomized clinical trial, comparing late coagulopathy in snakebitten patients treated with F(ab')2 with maintenance doses [F(ab')2/F(ab')2], or F(ab')2 with placebo maintenance doses [F(ab')2/placebo], versus Fab with maintenance doses [Fab/Fab]. The primary efficacy endpoint was coagulopathy (platelet count Fab/Fab cohort experienced late coagulopathy versus 4/39 (10.3%, p < 0.05) in the F(ab')2/F(ab')2 cohort and 2/38 (5.3%, p < 0.05) in the F(ab')2/placebo cohort. The lowest heterologous protein exposure was with F(ab')2/placebo. No serious adverse events were related to study drug. In each study arm, one patient experienced an acute serum reaction and one experienced serum sickness. In this study, management of coagulopathic Crotalinae envenomation with longer-half-life F(ab')2 antivenom, with or without maintenance dosing, reduced the risk of subacute coagulopathy and bleeding following treatment of envenomation.

  20. A screen for bacterial endosymbionts in the model organisms Tribolium castaneum, T. confusum, Callosobruchus maculatus, and related species.

    Science.gov (United States)

    Goodacre, Sara L; Fricke, Claudia; Martin, Oliver Y

    2015-04-01

    Reproductive parasites such as Wolbachia are extremely widespread amongst the arthropods and can have a large influence over the reproduction and fitness of their hosts. Undetected infections could thus confound the results of a wide range of studies that focus on aspects of host behavior, reproduction, fitness, and degrees of reproductive isolation. This potential problem has already been underlined by work investigating the incidence of Wolbachia infections in stocks of the model system Drosophila melanogaster. Here we survey a range of lab stocks of further commonly used model arthropods, focusing especially on the flour beetles Tribolium castaneum and Tribolium confusum, the cowpea weevil Callosobruchus maculatus and related species (Coleoptera: Tenebrionidae and Bruchidae). These species are widespread stored product pests so knowledge of infections with symbionts further has potential use in informing biocontrol measures. Beetles were assessed for infection with 3 known microbial reproductive parasites: Wolbachia, Rickettsia, Spiroplasma. Infections with some of these microbes were found in some of the lab stocks studied, although overall infections were relatively rare. The consequences of finding infections in these or other species and the type of previous studies likely to be affected most are discussed. © 2013 Institute of Zoology, Chinese Academy of Sciences.

  1. Structure of the omalizumab Fab

    Science.gov (United States)

    Jensen, Rasmus K.; Plum, Melanie; Tjerrild, Luna; Jakob, Thilo; Spillner, Edzard; Andersen, Gregers Rom

    2015-01-01

    Omalizumab is a humanized anti-IgE antibody that inhibits the binding of IgE to its receptors on mast cells and basophils, thus blocking the IgE-mediated release of inflammatory mediators from these cells. Omalizumab binds to the Fc domains of IgE in proximity to the binding site of the high-affinity IgE receptor Fc∊RI, but the epitope and the mechanisms and conformations governing the recognition remain unknown. In order to elucidate the molecular mechanism of its anti-IgE activity, the aim was to analyse the interaction of omalizumab with human IgE. Therefore, IgE Fc C∊2–4 was recombinantly produced in mammalian HEK-293 cells. Functionality of the IgE Fc was proven by ELISA and mediator-release assays. Omalizumab IgG was cleaved with papain and the resulting Fab was purified by ion-exchange chromatography. The complex of IgE Fc with omalizumab was prepared by size-exclusion chromatography. However, crystals containing the complex were not obtained, suggesting that the process of crystallization favoured the dissociation of the two proteins. Instead, two structures of the omalizumab Fab with maximum resolutions of 1.9 and 3.0 Å were obtained. The structures reveal the arrangement of the CDRs and the position of omalizumab residues known from prior functional studies to be involved in IgE binding. Thus, the structure of omalizumab provides the structural basis for understanding the function of omalizumab, allows optimization of the procedure for complex crystallization and poses questions about the conformational requirements for anti-IgE activity. PMID:25849503

  2. STECH, 3(3), S/NO 12, SEPTEMBER, 2014

    African Journals Online (AJOL)

    DR Nneka

    2014-09-12

    Sep 12, 2014 ... Observations on the Biology of Callosobruchus Maculatus. (Fab.) (Coleoptera: Bruchidae) under Ambient Laboratory ... carried out under ambient laboratory conditions. Pattern of oviposition, female adult longevity and F1 .... Effect of cowpea seed coat texture on the oviposition and progeny development of ...

  3. Journal of the Ghana Science Association - Vol 10, No 2 (2008)

    African Journals Online (AJOL)

    Susceptibility Of Five Cowpea (Vigna unguiculata) Varieties To Attack By Callosobruchus maculatus (Fab.) [Coleoptera: Bruchidae] · EMAIL FULL TEXT EMAIL FULL TEXT DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. JI Adam, PK Baidoo, 85-92. http://dx.doi.org/10.4314/jgsa.v10i2.18044 ...

  4. Meditope-Fab interaction: threading the hole.

    Science.gov (United States)

    Bzymek, Krzysztof P; Ma, Yuelong; Avery, Kendra N; Horne, David A; Williams, John C

    2017-12-01

    Meditope, a cyclic 12-residue peptide, binds to a unique binding side between the light and heavy chains of the cetuximab Fab. In an effort to improve the affinity of the interaction, it was sought to extend the side chain of Arg8 in the meditope, a residue that is accessible from the other side of the meditope binding site, in order to increase the number of interactions. These modifications included an n-butyl and n-octyl extension as well as hydroxyl, amine and carboxyl substitutions. The atomic structures of the complexes and the binding kinetics for each modified meditope indicated that each extension threaded through the Fab `hole' and that the carboxyethylarginine substitution makes a favorable interaction with the Fab, increasing the half-life of the complex by threefold compared with the unmodified meditope. Taken together, these studies provide a basis for the design of additional modifications to enhance the overall affinity of this unique interaction.

  5. Rapportage LTO FAB II 2008 : Functionele Agro Biodiversiteit

    NARCIS (Netherlands)

    Scheele, J.; Gurp, van H.; Alebeek, van F.A.N.; Belder, den E.; Elderson, J.

    2009-01-01

    Doel van het LTO FAB II project is een gebruiksklaar FAB concept te ontwikkelen voor een aantal ziekten en plagen in een aantal gewassen die op eenvoudige wijze door telers benut kan worden en voor de toepasser kostenneutraal zijn. Uitgangspunt in het FAB project is een evenwichtige balans tussen de

  6. FAB (Functionally Alert Behavior Strategies) to Improve Self-Control

    Science.gov (United States)

    Pagano, John

    2015-01-01

    This paper describes the FAB (Functionally Alert Behavior) Strategies approach to improve behavior in children and adolescents with complex behavioral challenges. FAB Strategies include evidence-based environmental adaptations, sensory modulation, positive behavioral support, and physical self-regulation strategies. FAB Strategies can be used by…

  7. Estabilidade da resistência de genótipos de caupi a Callosobruchus maculatus (Fabr. em gerações sucessivas

    Directory of Open Access Journals (Sweden)

    Lima Marcileyne Pessôa Leite de

    2002-01-01

    Full Text Available Callosobruchus maculatus (Fabr. (Coleoptera: Bruchidae é a principal praga do caupi (Vigna unguiculata L. armazenado em condições tropicais e subtropicais. Avaliaram-se a estabilidade da resistência e a capacidade de adaptação de C. maculatus a genótipos de caupi, durante seis gerações. Utilizou-se o teste sem chance de escolha, em delineamento experimental inteiramente casualizado, em esquema fatorial, com dez tratamentos, seis gerações do inseto e cinco repetições. Cada repetição constou de 30 grãos infestados com dois casais da praga. O número de ovos/fêmea diferiu entre os genótipos de caupi apenas na sexta geração, e entre as gerações em BR14-Mulato, Bico de Pato, TE90-180-3E e TE87-98-8G. A viabilidade de ovos diferiu entre as gerações em BR17-Gurguéia, BR14-Mulato, IT89KD-260 e IT89KD-245, e entre os genótipos nas terceira, quinta e sexta gerações. Os genótipos diferiram entre e dentro das gerações, em relação à duração e a viabilidade da fase imatura. Observou-se redução na emergência, especialmente em IT89KD-245, IT89KD-260, CNC 0434, Bico de Pato, TE90-180-10F e BR14-Mulato, provavelmente devido a substâncias químicas presentes nos grãos que afetaram a sobrevivência dos insetos, ao longo das gerações. Os genótipos IT89KD-245 e IT89KD-260 comportaram-se como moderadamente resistentes. Não se observou adaptação de C. maculatus aos genótipos, mantendo-se a resistência estável através das gerações.

  8. FabH Mutations Confer Resistance to FabF-Directed Antibiotics in Staphylococcus aureus

    OpenAIRE

    Parsons, Joshua B.; Yao, Jiangwei; Frank, Matthew W.; Rock, Charles O.

    2014-01-01

    Delineating the mechanisms for genetically acquired antibiotic resistance is a robust approach to target validation and anticipates the evolution of clinical drug resistance. This study defines a spectrum of mutations in fabH that render Staphylococcus aureus resistant to multiple natural products known to inhibit the elongation condensing enzyme (FabF) of bacterial type II fatty acid synthesis. Twenty independently isolated clones resistant to platensimycin, platencin, or thiolactomycin were...

  9. Controlled Fab installation onto polymeric micelle nanoparticles for tuned bioactivity

    Science.gov (United States)

    Chen, Shaoyi; Florinas, Stelios; Teitgen, Abigail; Xu, Ze-Qi; Gao, Changshou; Wu, Herren; Kataoka, Kazunori; Cabral, Horacio; Christie, R. James

    2017-12-01

    Antibodies and antigen-binding fragments (Fabs) can be used to modify the surface of nanoparticles for enhanced target binding. In our previous work, site-specific conjugation of Fabs to polymeric micelles using conventional methods was limited to approximately 30% efficiency, possibly due to steric hindrance related to macromolecular reactants. Here, we report a new method that enables conjugation of Fabs onto a micelle surface in a controlled manner with up to quantitative conversion of nanoparticle reactive groups. Variation of (i) PEG spacer length in a heterofunctionalized cross-linker and (ii) Fab/polymer feed ratios resulted in production of nanoparticles with a range of Fab densities on the surface up to the theoretical maximum value. The biological impact of variable Fab density was evaluated in vitro with respect to cell uptake and cytotoxicity of a drug-loaded (SN38) targeted polymeric micelle bearing anti-EphA2 Fabs. Fab conjugation increased cell uptake and potency compared with non-targeted micelles, although a Fab density of 60% resulted in decreased uptake and potency of the targeted micelles. Altogether, our findings demonstrate that conjugation strategies can be optimized to allow control of Fab density on the surface of nanoparticles and also that Fab density may need to be optimized for a given cell-surface target to achieve the highest bioactivity.

  10. Fungi associated to Platypus cylindrus Fab. (Coleoptera: Platypodidae in cork oak Fungos associados ao insecto Platypus cylindrus Fab. (Coleoptera: Platypodidae em sobreiro

    Directory of Open Access Journals (Sweden)

    Joana Henriques

    2009-12-01

    Full Text Available Platypus cylindrus is a pest that since the 80’s of the last century has been considered a cork oak mortality agent in Portugal. It is an ambrosia beetle that establishes complex symbioses with fungi whose role in the insect-fungus-host interaction has not been completely clarified. In order to characterize P. cylindrus associated micoflora in Portugal, fungi were isolated from different beetle organs and from its galleries in cork oak trees. Fungi of the genera Acremonium, Aspergillus, Beauveria, Botrytis, Chaetomium, Fusarium, Geotrichum, Gliocladium, Nodulisporium, Paecilomyces, Penicillium, Raffaelea, Scytalidium, Trichoderma and of the order Mucorales were identified. An actinomycete of the genus Streptomyces was also identified. Some of these genera were related for the first time to this interaction. In the present work the isolated fungi are characterized and their contribution for beetle population establishment and tree weakness is discussed.Platypus cylindrus é uma praga que desde os anos 80 do século passado tem sido referida como agente de mortalidade do sobreiro em Portugal. É um insecto ambrósia que estabelece simbioses complexas com fungos cujo papel não está completamente esclarecido na interacção insecto-fungo-sobreiro. Com o objectivo de caracterizar a micoflora associada a P. cylindrus em Portugal foram efectuados isolamentos a partir de diferentes órgãos do insecto e suas galerias em sobreiro. Identificaram-se fungos dos géneros Acremonium, Aspergillus, Beauveria, Botrytis, Chaetomium, Fusarium, Geotrichum, Gliocladium, Nodulisporium, Paecilomyces, Penicillium, Raffaelea, Scytalidium, Trichoderma e da ordem Mucorales. Foi igualmente identificado um actinomiceta do género Streptomyces. Alguns destes géneros são referidos pela primeira vez nesta interacção. No presente trabalho caracterizam-se os fungos isolados e discute-se a sua contribuição para o estabelecimento das populações do insecto e enfraquecimento das árvores.

  11. PSMA-targeted bispecific Fab conjugates that engage T cells.

    Science.gov (United States)

    Patterson, James T; Isaacson, Jason; Kerwin, Lisa; Atassi, Ghazi; Duggal, Rohit; Bresson, Damien; Zhu, Tong; Zhou, Heyue; Fu, Yanwen; Kaufmann, Gunnar F

    2017-12-15

    Bioconjugate formats provide alternative strategies for antigen targeting with bispecific antibodies. Here, PSMA-targeted Fab conjugates were generated using different bispecific formats. Interchain disulfide bridging of an αCD3 Fab enabled installation of either the PSMA-targeting small molecule DUPA (SynFab) or the attachment of an αPSMA Fab (BisFab) by covalent linkage. Optimization of the reducing conditions was critical for selective interchain disulfide reduction and good bioconjugate yield. Activity of αPSMA/CD3 Fab conjugates was tested by in vitro cytotoxicity assays using prostate cancer cell lines. Both bispecific formats demonstrated excellent potency and antigen selectivity. Copyright © 2017. Published by Elsevier Ltd.

  12. Bringing optics to Fab Labs in Europe

    Science.gov (United States)

    Adam, Aurèle; Zuidwijk, Thim; Urbach, Paul

    2017-08-01

    The Optics Group of Delft University of Technology plays a major role in teaching optics to bachelor and master students. In addition, the group has a long record of introducing, demonstrating and teaching optics to quite diverse groups of people from outside of the university. We will describe some of these activities and focus on a recently started project funded by the European Commission called Phablabs 4.0, which aims to bring photonics to European Fab labs.

  13. Machine learning for fab automated diagnostics

    Science.gov (United States)

    Giollo, Manuel; Lam, Auguste; Gkorou, Dimitra; Liu, Xing Lan; van Haren, Richard

    2017-06-01

    Process optimization depends largely on field engineer's knowledge and expertise. However, this practice turns out to be less sustainable due to the fab complexity which is continuously increasing in order to support the extreme miniaturization of Integrated Circuits. On the one hand, process optimization and root cause analysis of tools is necessary for a smooth fab operation. On the other hand, the growth in number of wafer processing steps is adding a considerable new source of noise which may have a significant impact at the nanometer scale. This paper explores the ability of historical process data and Machine Learning to support field engineers in production analysis and monitoring. We implement an automated workflow in order to analyze a large volume of information, and build a predictive model of overlay variation. The proposed workflow addresses significant problems that are typical in fab production, like missing measurements, small number of samples, confounding effects due to heterogeneity of data, and subpopulation effects. We evaluate the proposed workflow on a real usecase and we show that it is able to predict overlay excursions observed in Integrated Circuits manufacturing. The chosen design focuses on linear and interpretable models of the wafer history, which highlight the process steps that are causing defective products. This is a fundamental feature for diagnostics, as it supports process engineers in the continuous improvement of the production line.

  14. Biology of Pityophthorus pulchellus tuberculatus (Coleoptera: Curculionidae: Scolytinae) in Lodgepole Pine in Northern Idaho.

    Science.gov (United States)

    Furniss, Malcolm M; Kegley, Sandra J

    2018-04-19

    The twig beetle, Pityophthorus pulchellus tuberculatus Eichhoff, infests dead branches of pines in western United States and Canada, including lodgepole pine, Pinus contorta Douglas, in northern Idaho. Adult broods overwintered in their host and emerged and colonized new hosts in late April. Males initiated galleries and were joined by up to seven females, each of which constructed an egg gallery radiating from a central chamber. Galleries had an average of 4.7 egg niches each with an egg that was large relative to the mother beetle. Two larval instars were recognized. Dentition of larval mandibles differed in shape from that in literature. Mature larvae pupated either in a cell excavated on the wood surface or in a cell below the wood surface. First-generation adults mined extensively in the inner bark and wood before emerging to infest new trees in late June. Their progeny became adults beginning in early August and likewise mined and fed on the inner bark and wood before overwintering. Predacious beetles present as larvae in the galleries included Enoclerus lecontei (Wolcott) (Coleoptera: Cleridae) and Lasconotus sp. (Coleoptera: Zopheridae). Parasitoid Hymenoptera reared from infested trees were Cosmophorus pityophthori Rohwer (Braconidae), Phasmidiasta n. sp. (Braconidae), Spathius sp. (Braconidae), Acerocephala n. sp. (Pteromalidae), Metacolus fasciatus Girault (Pteromalidae), Rhaphitelus maculatus Walker (Pteromalidae), Rhopalicus sp. (Pteromalidae), and an unidentified pteromalid.

  15. Radioiodinated iodobenzoyl conjugates of a monoclonal antibody Fab fragment. In vivo comparisons with chloramine-T-labeled Fab

    International Nuclear Information System (INIS)

    Wilbur, D.S.; Hadley, S.W.; Grant, L.M.; Hylarides, M.D.

    1991-01-01

    A comparative investigation of the biodistributions of radioiodinated p- and m-iodobenzoyl conjugates of a monoclonal antibody Fab fragment, NR-LU-10 Fab, and the same antibody Fab fragment radioiodinated by the chloramine-T (ChT) method has been carried out in mice. Coinjected, dual-isotope studies in athymic mice with tumor xenografts have demonstrated that there are only minor differences in the in vivo distributions of the iodobenzoyl-labeled Fabs, except in the excretory organs, kidneys, and intestines, where major differences were observed. Similarly, coinjection of either the p-iodobenzoyl or m-iodobenzoyl conjugate of NR-LU-10 Fab with the Fab radioiodinated with ChT/radioiodide into BALB/c mice provided additional data that indicated that the two iodobenzoyl conjugates distributed similar in a number of selected tissues. The tissue-distribution differences of the regioisomeric iodobenzoyl conjugates in relation to the ChT-radioiodinated Fab were large for the stomach and neck, consistent with previous studies. The most notable difference between the two iodobenzoyl conjugates was the kidney activity, where the m-iodobenzoyl conjugate was similar to the directly labeled Fab, but the p-iodobenzoyl-conjugated Fab was higher by nearly a factor of 2

  16. UVB-induced gene expression in the skin of Xiphophorus maculatus Jp 163 B☆

    Science.gov (United States)

    Yang, Kuan; Boswell, Mikki; Walter, Dylan J.; Downs, Kevin P.; Gaston-Pravia, Kimberly; Garcia, Tzintzuni; Shen, Yingjia; Mitchell, David L.; Walter, Ronald B.

    2014-01-01

    Xiphophorus fish and interspecies hybrids represent long-standing models to study the genetics underlying spontaneous and induced tumorigenesis. The recent release of the Xiphophorus maculatus genome sequence will allow global genetic regulation studies of genes involved in the inherited susceptibility to UVB-induced melanoma within select backcross hybrids. As a first step toward this goal, we report results of an RNA-Seq approach to identify genes and pathways showing modulated transcription within the skin of X. maculatus Jp 163 B upon UVB exposure. X. maculatus Jp 163 B were exposed to various doses of UVB followed by RNA-Seq analysis at each dose to investigate overall gene expression in each sample. A total of 357 genes with a minimum expression change of 4-fold (p-adj fish skin to UVB exposure. PMID:24556253

  17. The mimetic repertoire of the spotted bowerbird Ptilonorhynchus maculatus

    Science.gov (United States)

    Kelley, Laura A.; Healy, Susan D.

    2011-06-01

    Although vocal mimicry in songbirds is well documented, little is known about the function of such mimicry. One possibility is that the mimic produces the vocalisations of predatory or aggressive species to deter potential predators or competitors. Alternatively, these sounds may be learned in error as a result of their acoustic properties such as structural simplicity. We determined the mimetic repertoires of a population of male spotted bowerbirds Ptilonorhynchus maculatus, a species that mimics predatory and aggressive species. Although male mimetic repertoires contained an overabundance of vocalisations produced by species that were generally aggressive, there was also a marked prevalence of mimicry of sounds that are associated with alarm such as predator calls, alarm calls and mobbing calls, irrespective of whether the species being mimicked was aggressive or not. We propose that it may be the alarming context in which these sounds are first heard that may lead both to their acquisition and to their later reproduction. We suggest that enhanced learning capability during acute stress may explain vocal mimicry in many species that mimic sounds associated with alarm.

  18. Een inventarisatie naar de bekendheid van Functionele Agrobiodiversiteit (FAB) en de mogelijkheden om FAB met andere agroranden te combineren

    NARCIS (Netherlands)

    Geerts, R.H.E.M.; Meerburg, B.G.

    2011-01-01

    Op dit moment zijn er regio's waar het FAB concept al breed wordt toegepast en goed is ingebed (West-Brabant/Zeeland), waar het in opmars is (Flevoland en Zuid-Holland) en enkele regio's waar het FAB gedachtegoed slechts matig aanwezig is (Groningen / Drenthe).

  19. Efficacy of IgG, Fab, and F(ab')2 fragments of horse antivenom in the ...

    African Journals Online (AJOL)

    The ability of horse antivenoms, consisting of immunoglobulin G (IgG) and its fragments F(ab')2 and Fab were comparatively studied in mice to neutralize several effects of Cerastes cerastes venom. The three antivenoms were produced from the same batch of hyperimmune horse plasma. Neutralization was only partial ...

  20. The cosmology of the Fab-Four

    International Nuclear Information System (INIS)

    Copeland, Edmund J.; Padilla, Antonio; Saffin, Paul M.

    2012-01-01

    We have recently proposed a novel self tuning mechanism to alleviate the famous cosmological constant problem, based on the general scalar tensor theory proposed by Horndeski. The self-tuning model ends up consisting of four geometric terms in the action, with each term containing a free potential function of the scalar field; the four together being labeled as the Fab-Four. In this paper we begin the important task of deriving the cosmology associated with the Fab-Four Lagrangian. Performing a phase plane analysis of the system we are able to obtain a number of fixed points for the system, with some remarkable new solutions emerging from the trade-off between the various potentials. As well as obtaining inflationary solutions we also find conventional radiation/matter-like solutions, but in regimes where the energy density is dominated by a cosmological constant, and where we do not have any explicit forms of radiation or matter. Stability conditions for matter solutions are obtained and we show how it is possible for there to exist an extended period of 'matter domination' opening up the possibility that we can generate cosmological structures, and recover a consistent cosmology even in the presence of a large cosmological constant

  1. The damage caused by Callosobruchus maculatus on cowpea grains is dependent on the plant genotype.

    Science.gov (United States)

    Torres, Elida Barros; Nóbrega, Rafaela S A; Fernandes-Júnior, Paulo Ivan; Silva, Luciana Barboza; Dos Santos Carvalho, Gabriel; Marinho, Rita de Cassia Nunes; Pavan, Bruno E

    2016-09-01

    Beans from cowpea cultivars fertilized with mineral N or inoculated with various rhizobium strains may contain different nitrogen concentrations and nitrogen metabolite composition, which affects the beans' defense mechanisms against pests. In this study, the population growth of Callosobruchus maculatus reared on beans from four cowpea cultivars fertilized with different nitrogen sources was evaluated. The factors tested were beans from four cowpea cultivars and seven different nitrogen sources: mineral N fertilization, inoculation with five strains of symbiotic diazotrophic bacteria, and soil nitrogen (absolute control). BRS Tapaihum and BRS Acauã cultivars had lower cumulative emergence and instantaneous rate of population growth of the insects compared with other cultivars, indicating antixenosis resistance against C. maculatus. Inoculation of BRS Acauã cultivar with the diazotrophic bacteria strain BR 3299 resulted in higher mortality of C. maculatus. For BRS Tapaihum cultivar, inoculation with diazotrophic bacteria strains BR3267, BR 3262 and BR 3299, and nitrogen fertilization resulted in higher mortality among C. maculatus. BRS Tapaihum and BRS Acauã cultivars showed the lowest cumulative insect emergence and instantaneous rates of population growth, and the highest insect mortality, mainly when the grains were obtained from plants inoculated with rhizobial strains. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  2. The interactive effect of entomopathogenic nematode, Steinernema carpocapsae (Weiser) and gamma radiation ori callosobruchus maculatus (F.) and their biochemical Effects

    International Nuclear Information System (INIS)

    EI-Orabi, M.N.; Slwfei, D.M.Y.; Amer, A.M.

    2007-01-01

    Cowpea weevil, Callosobruchus maculatus (F.) adults were infected with entomopathogenic nematodes, Steinernema carpocapsae (Weiser) for evaluating their potential for suppressing this insect pest. Morality percentage of un-irradiated C. maculatus infected with irradiated nematodes decreased at higher doses of gamma irradiation. The results generally Indicated that the reduction of mortality was directly related to gamma irradiation. The results indicated that infestation intensity decreased at higher doses of gamma irradiation when irradiated nematodes infected the irradiated or non-irradiated C. maculatus. Protein content of irradiated C. maculatus adults infected with nematodes showed a slight decrease by increasing radiation dose and time elapsed from infection. Comparing the protein bands at different time intervals for all tested dosage used showed absence of some bands and reduction in the intensity of the others

  3. Semiconductor industry wafer fab exhaust management

    CERN Document Server

    Sherer, Michael J

    2005-01-01

    Given the myriad exhaust compounds and the corresponding problems that they can pose in an exhaust management system, the proper choice of such systems is a complex task. Presenting the fundamentals, technical details, and general solutions to real-world problems, Semiconductor Industry: Wafer Fab Exhaust Management offers practical guidance on selecting an appropriate system for a given application. Using examples that provide a clear understanding of the concepts discussed, Sherer covers facility layout, support facilities operations, and semiconductor process equipment, followed by exhaust types and challenges. He reviews exhaust point-of-use devices and exhaust line requirements needed between process equipment and the centralized exhaust system. The book includes information on wet scrubbers for a centralized acid exhaust system and a centralized ammonia exhaust system and on centralized equipment to control volatile organic compounds. It concludes with a chapter devoted to emergency releases and a separ...

  4. How fabulous is Fab 5 cosmology?

    International Nuclear Information System (INIS)

    Linder, Eric V.

    2013-01-01

    Extended gravity origins for cosmic acceleration can solve some fine tuning issues and have useful characteristics, but generally have little to say regarding the cosmological constant problem. Fab 5 gravity can be ghost free and stable, have attractor solutions in the past and future, and possess self tuning that solves the original cosmological constant problem. Here we show however it does not possess all these qualities at the same time. We also demonstrate that the self tuning is so powerful that it not only cancels the cosmological constant but also all other energy density, and we derive the scalings of its approach to a renormalized de Sitter cosmology. While this strong cancellation is bad for the late universe, it greatly eases early universe inflation

  5. How fabulous is Fab 5 cosmology?

    Energy Technology Data Exchange (ETDEWEB)

    Linder, Eric V., E-mail: evlinder@lbl.gov [Berkeley Center for Cosmological Physics and Berkeley Lab, University of California, Berkeley, CA, 94720 (United States)

    2013-12-01

    Extended gravity origins for cosmic acceleration can solve some fine tuning issues and have useful characteristics, but generally have little to say regarding the cosmological constant problem. Fab 5 gravity can be ghost free and stable, have attractor solutions in the past and future, and possess self tuning that solves the original cosmological constant problem. Here we show however it does not possess all these qualities at the same time. We also demonstrate that the self tuning is so powerful that it not only cancels the cosmological constant but also all other energy density, and we derive the scalings of its approach to a renormalized de Sitter cosmology. While this strong cancellation is bad for the late universe, it greatly eases early universe inflation.

  6. Rasch Analysis of the Fullerton Advanced Balance (FAB) Scale

    OpenAIRE

    Klein, Penelope J.; Fiedler, Roger C.; Rose, Debra J.

    2011-01-01

    Purpose: This cross-sectional study explores the psychometric properties and dimensionality of the Fullerton Advanced Balance (FAB) Scale, a multi-item balance test for higher-functioning older adults.

  7. A Case Study of a High School Fab Lab

    Science.gov (United States)

    Lacy, Jennifer E.

    This dissertation examines making and design-based STEM education in a formal makerspace. It focuses on how the design and implementation of a Fab Lab learning environment and curriculum affect how instructors and students see themselves engaging in science, and how the Fab Lab relates to the social sorting practices that already take place at North High School. While there is research examining design-based STEM education in informal and formal learning environments, we know little about how K-12 teachers define STEM in making activities when no university or museum partnership exists. This study sought to help fill this gap in the research literature. This case study of a formal makerspace followed instructors and students in one introductory Fab Lab course for one semester. Additional observations of an introductory woodworking course helped build the case and set it into the school context, and provided supplementary material to better understand the similarities and differences between the Fab Lab course and a more traditional design-based learning course. Using evidence from observational field notes, participant interviews, course materials, and student work, I found that the North Fab Lab relies on artifacts and rhetoric symbolic of science and STEM to set itself apart from other design-based courses at North High School. Secondly, the North Fab Lab instructors and students were unable to explain how what they were doing in the Fab Lab was science, and instead relied on vague and unsupported claims related to interdisciplinary STEM practices and dated descriptions of science. Lastly, the design and implementation of the Fab Lab learning environment and curriculum and its separation from North High School's low tech, design-based courses effectively reinforced social sorting practices and cultural assumptions about student work and intelligence.

  8. Bioactivity of Lantana camara L. essential oil against Callosobruchus maculatus (Fabricius Bioactividad de aceite esencial de Lantana camara L. contra Callosobruchus maculatus (Fabricius

    Directory of Open Access Journals (Sweden)

    Nooshin Zandi-Sohani

    2012-12-01

    Full Text Available Lantana camara L. is a widespread plant species mostly native to subtropical and tropical regions of the world. In this study, insecticidal and repellent activities of L. camara essential oil were evaluated against Callosobruchus maculatus (Fabricius (Col: Bruchidae. Analysis of chemical composition by gas chromatography/mass spectrometry (GC/MS showed high amounts of sesquiterpenes, mainly α-humelene (23.3% and cis-caryophyllene (16.2%. The results showed that the essential oil of L. camara has strong repellent activity against adults of C. maculatus at all tested concentrations. After 2 and 4 h, 97.4 and 100% repellency was seen at highest concentrations of 0.4 μL cm-2, respectively. Moreover, the oil was found to be toxic to adults when applied by fumigation. Responses varied with the gender of insect and exposure time. The LC50 values were 282.7 and 187.9 μL L-1 for females and males, respectively. An increase in the exposure time from 3 to 24 h caused increasing in mortality from 23.6 to 100% in males and from 14.1 to 97.1% in females, at highest concentration (1160 μL L-1. According to these results, L. camara essential oil may be useful as an alternative for bean protection against C. maculatus.Lantana camara L. es una especie vegetal nativa de las regiones subtropicales y tropicales del mundo. En este estudio se evaluaron las actividades insecticida y repelente del aceite esencial de L. camara contra Callosobruchus maculatus (Fabricius. El análisis de la composición volátil de este aceite esencial mediante cromatografía de gas/ espectrometría de masa (GC/MS demostró la presencia de elevadas cantidades de sesquiterpenos, principalmente α-humuleno (23.3% y ci's-cariofileno (16.2%. Los resultados obtenidos demostraron que el aceite esencial de L. camara tiene una elevada actividad repelente contra los adultos de C. maculatus a todas las concentraciones ensayadas. Después de 2 y 4 h la concentración de 0.4 μL cm-2 causó 97

  9. Camallanus maculatus n. sp. (Nematoda: Camallanidae in an ornamental fish Xiphophorus maculatus (Osteichthyes: Poeciliidae cultivated in São Paulo State, Brazil Camallanus maculatus n. sp. (Nematoda: Camallanidae em peixe ornamental Xiphophorus maculatus (Osteichthyes: Poeciliidae cultivado no estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    M.L. Martins

    2007-10-01

    Full Text Available The present study describes Camallanus maculatus n. sp. (Nematoda: Camallanidae from Xiphophorus maculatus (Osteichthyes: Poecilidae collected in an ornamental fish farm in São Paulo State, Brazil. Thirty-three out of 40 examined fish were parasitized (82% prevalence with mean intensity of 2.8 (1-20. Male specimens were 3,500µm total length and females 7,100µm total length, provided with orange-brown buccal capsule and two developed tridents. Males showed two unequal spicules, gubernaculum absent and females with vulva situated in the middle of the body. The main characteristics were the sleekly longitudinal ridges in the buccal capsule in both sexes, enlargement at the anterior end of the smaller spicule, the number of genital papillae, longer muscular and glandular portion of esophagus and round tail in female.Descreve-se Camallanus maculatus n. sp. (Nematoda: Camallanidae em Xiphophorus maculatus (Osteichthyes: Poecilidae capturado em uma piscicultura ornamental no estado de São Paulo. De 40 peixes examinados, 33 estavam parasitados (prevalência de 82% cuja intensidade média foi de 2,8 (1-20. Os machos apresentaram 3.500µm e as fêmeas com 7,100µm de comprimento total, providos de uma cápsula bucal laranja-castanho e dois tridentes. Observaram-se machos com espículos desiguais, gubernáculo ausente e fêmeas com vulva situada na metade do corpo. As principais características foram as estrias longitudinais lisas em cada valva da cápsula bucal em ambos os sexos, o espículo menor com alargamento na extremidade anterior, o número de papilas genitais, as porções muscular e glandular do esôfago longas e a cauda longa e arredondada na fêmea.

  10. Sensitivity of the different developmental stages of Callosobruchus maculatus to gamma irradiation

    International Nuclear Information System (INIS)

    Olaifa, J.I.; Adebayo, T.A.; Adesanmi, C.A.; Salau, A.M.

    1990-01-01

    Life stages of the cowpea seed beetle (Callosobruchus maculatus F.) were γ irradiated from a 60 Co source. Arrested development was observed at dosages of 1.0, 4.0, 10.0 and 10.0 Gy in the egg, 1st, 2nd, 3rd and 4th larvae stages respectively while 4.0 Gy induced sterility. Longevity of 2-day old adults was not affected by irradiation of 100 Gy. Preservation of cowpeas by irradiation is discussed. (author)

  11. Alterations in an indium-111 Fab' under conditions of utilization

    International Nuclear Information System (INIS)

    Halpern, S.E.; Sudora, E.; Tarburton, J.P.; Hagar, P.

    1992-01-01

    This study was conducted to investigate alterations that occur in an indium/111 Fab' of a monoclonal antibody following its in vivo administration. Patients were infused with 111 In-Fab' of the monoclonal antibody ZCE-025. Serum and urine specimens were collected from these pateients. Starting materials, serum, urine and controls samples were studied by electrophoresis. Animal distribution studies were performed in normal Balb/c mice and, in some cases, nude mice bearing a carcinoembryonic antigen (CEA)/producing human colon tumour since the antibody targets CEA. The studies indicated that the molecule circulated almost totally intact for at least 4 h and to a considerable extent for 24 h, with some evidence for in vivo fragmentation by 24 h. Evidence was also obtained suggesting the formation of a high molecular weight species in some patients. Shortly after infusion, some of the 111 In in the urine appeared as the intact Fab', but within hours the majority migrated electro-phoretically as low molecular weight species. We conclude that while the majority of the 111 In-Fab' of this particular antibody remains intact and immunoreactive following its administration, the molecule is structurally changed to some degree shortly after its infusion into humans. Since each monoclonal antibody is unique, the degree and rapidity of degradation of its Fab' in vivo could vary markedly from the above and possibly adversely effect its utility as a radiopharmaceutical. (orig.)

  12. 1825 (Coleoptera: Cerambycidae

    Directory of Open Access Journals (Sweden)

    Márcia d´Avila

    2006-01-01

    Full Text Available The erva-mate (Ilex paraguariensis A.St.-Hil. has a social, cultural and economic importance in the southern states of Brazil. The pure stands of this culture was responsible for the increase of many species of insects. Hedypathes betulinus (Klug, 1825 (Coleoptera: Cerambycidae is considered the main pest from an economic viewpoint because of its difficult control and potential for damage. The larval phase occurs inside the twings and trunks, what makes more difficult to deal with its detention and management. Cultural and mechanical management are the most indicated, such as collection of adults, prunning and burning of plant parts damaged by the insect, balanced nutrition, adequate plant density and maintenance of areas with native vegetation or also the introduction of policulture. These strategies may increase the agroecossystem balance and thus a reduction of the insect-pest to an aceptable level. Therefore, the objective of this study was to assemble and the discuss the information on the bioecology and management of erva-mate borer.

  13. 20 CFR 30.316 - How does the FAB issue a final decision on a claim?

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false How does the FAB issue a final decision on a... Adjudicatory Process Hearings and Final Decisions on Claims § 30.316 How does the FAB issue a final decision on... waives any objections to all or part of the recommended decision, the FAB may issue a final decision...

  14. 21 CFR 866.5520 - Immunoglobulin G (Fab fragment specific) immunological test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Immunoglobulin G (Fab fragment specific... Test Systems § 866.5520 Immunoglobulin G (Fab fragment specific) immunological test system. (a) Identification. An immunoglobulin G (Fab fragment specific) immunological test system is a device that consists...

  15. Albizia lebbeck Seed Coat Proteins Bind to Chitin and Act as a Defense against Cowpea Weevil Callosobruchus maculatus.

    Science.gov (United States)

    Silva, Nadia C M; De Sá, Leonardo F R; Oliveira, Eduardo A G; Costa, Monique N; Ferreira, Andre T S; Perales, Jonas; Fernandes, Kátia V S; Xavier-Filho, Jose; Oliveira, Antonia E A

    2016-05-11

    The seed coat is an external tissue that participates in defense against insects. In some nonhost seeds, including Albizia lebbeck, the insect Callosobruchus maculatus dies during seed coat penetration. We investigated the toxicity of A. lebbeck seed coat proteins to C. maculatus. A chitin-binding protein fraction was isolated from seed coat, and mass spectrometry showed similarity to a C1 cysteine protease. By ELM program an N-glycosylation interaction motif was identified in this protein, and by molecular docking the potential to interact with N-acetylglucosamine (NAG) was shown. The chitin-binding protein fraction was toxic to C. maculatus and was present in larval midgut and feces but not able to hydrolyze larval gut proteins. It did not interfere, though, with the intestinal cell permeability. These results indicate that the toxicity mechanism of this seed coat fraction may be related to its binding to chitin, present in the larvae gut, disturbing nutrient absorption.

  16. Thrombus imaging with indium-111 and iodine-131-labeled fibrin-specific monoclonal antibody and its F(ab')2 and Fab fragments

    International Nuclear Information System (INIS)

    Rosebrough, S.F.; Grossman, Z.D.; McAfee, J.G.

    1988-01-01

    We have previously reported successful imaging of fresh (2-4 hr old) and aged (1-5 days old) canine thrombi with 131 I-labeled intact monoclonal antibody (MAb) specific for fibrin. We now report thrombus imaging with 131 I-labeled F(ab')2 and Fab and 111 In-labeled intact MAb, F(ab')2, and Fab. Indium-111-labeled F(ab')2 proved to be the best imaging agent due to less nonspecific binding in the liver than whole IgG. Image quality was improved by the higher administered dose permissible with 111 In and its better physical characteristics for imaging, compared to 131 I. Immunofluorescence of fresh human histologic sections showed intact MAb and F(ab')2 binding to thrombi, pulmonary emboli, and atherosclerotic plaques, strengthening the feasibility of clinical thrombus imaging

  17. Pollution prevention opportunity assessment for MicroFab and SiFab facilities at Sandia National Laboratories.

    Energy Technology Data Exchange (ETDEWEB)

    Gerard, Morgan Evan

    2011-12-01

    This Pollution Prevention Opportunity Assessment (PPOA) was conducted for the MicroFab and SiFab facilities at Sandia National Laboratories/New Mexico in Fiscal Year 2011. The primary purpose of this PPOA is to provide recommendations to assist organizations in reducing the generation of waste and improving the efficiency of their processes and procedures. This report contains a summary of the information collected, the analyses performed, and recommended options for implementation. The Sandia National Laboratories Environmental Management System (EMS) and Pollution Prevention (P2) staff will continue to work with the organizations to implement the recommendations.

  18. Direct labelling of monomeric antibody fragments Fab' with 99mTc

    International Nuclear Information System (INIS)

    Li Jun; Wang Shizhen; Yang Ziyi

    1994-01-01

    Direct labelling method and conditions of monomeric antibody Fab' with 99m Tc were investigated. Polyclonal antibody IgG was digested with ficin to produce dimeric fragments F(ab') 2 , which was subsequently reduced to monomeric fragments Fab' with 2-mercaptoethylamine. Finally, Fab' was incubated with sodium gluconate (Sn(II)) kit solution and 99m TcO 4 - eluted at room temperature to form 99m Tc-Fab'. The labelling efficiency was 85%-95%. The stability of labelled products was satisfactory and the elimination rate was faster than 99m Tc-IgG

  19. 20 CFR 30.319 - May a claimant request reconsideration of a final decision of the FAB?

    Science.gov (United States)

    2010-04-01

    ... final decision of the FAB? 30.319 Section 30.319 Employees' Benefits OFFICE OF WORKERS' COMPENSATION... reconsideration of a final decision of the FAB? (a) A claimant may request reconsideration of a final decision of the FAB by filing a written request with the FAB within 30 days from the date of issuance of such...

  20. COMPARATIVE STUDY OF THE TOXICITY OF OILS FROM SEEDS OF Citrullus colocynthis AND Citrullus vulgaris ON LARVAE OF Dermestes Maculatus

    OpenAIRE

    Akpotu, Onoriode Joseph; Oniye, Sonnie Johnson; Abolude, David; Yusuf, Abdullateef

    2016-01-01

    Studywas carried out in the Fisheries Research Laboratory, Ahmadu Bello Universityto evaluate the effect of Citrulluscolocynthis (Eguisi) and Citrullusvulgaris (Watermelon) seed oils on the larvae of Dermestes maculatus, an important pest of smoke-dried Clarias gariepinus (African catfish).The effect of the oils at different tested concentrations against the larvae ofD. maculatus was dose dependent as3.33% and 81.11% mortality for C.colocynthis while 2.22% and 91.11% for C.vulgaris were recor...

  1. Description of Acleotrema maculatus sp. nov. (Monogenea: Diplectanidae infecting the spotted coral grouper Plectropomus maculatus (F:Serranidea from the Red Sea and its histopathological impact

    Directory of Open Access Journals (Sweden)

    Kareem Morsy

    2014-09-01

    Full Text Available Acleotrema maculatus sp. nov. (Monogenea: Diplectanidae was described from the gills of The spotted coral grouper Plectropomus maculatus (F:Serranidea, Forsskal, 1775. Fish were collected from boot landing sites and fishermen at different water locations along the Red Sea at Hurghada City, Egypt. The morphology and morphometric characterization of the recovered worms were described by means of light microscopy. Eight (53.3% out 15 specimens of P. maculatus were infected. Most of the infected fish had very pale gills. Morphologically, the adult worm of A. maculatus sp. nov., possessed a body which was elongated, fusiform with a total length 0.86–0.90 (0.88 ± 0.02 mm, and a maximum width 0.09–0.13 (0.11 ± 0.02 mm at the level of ovary. Haptor, broad, differentiated from the rest of the body, measured 0.04–0.08 (0.06 ± 0.02 mm and provided with continuous rows of squamodiscs. Two pairs of lateral hamuli, three bars and 14 marginal hooklets were also observed. Lateral (dorsal bars two, stout, dumbbell-shaped, measured 0.052–0.056 (0.054 ± 0.002 mm in length. Ventral bar slender, with transverse groove, measured 0.09–0.11 (0.10 ± 0.01 mm in length. Ventral hamuli measured 0.02–0.06 (0.04 ± 0.02 mm in total length with an outer root that was very long measured 0.016–0.02 (0.018 ± 0.002, stout, slightly notched at broad proximal end; inner root was conical and measured 0.013–0.017 (0.015 ± 0.002 mm, with shaft measured 0.025–0.029 (0.027 ± 0.002 mm and point length 0.007–0.009 (0.008 ± 0.002 mm. Dorsal hamuli measured 0.034–0.038 (0.036 ± 0.002 in total length; base large, stout, with only lateral rudiment of roots; blade and point were long and curved with shaft length measured 0.024–0.028 (0.026 ± 0.002 mm and 0.02–0.06 (0.04 ± 0.02 mm point length. The worm is hermaphrodite, male copulatory organ measured 0.058–0.062 (0.060 ± 0.002 mm in length with a sclerotized part composed of

  2. Improved Fab presentation on phage surface with the use of molecular chaperone coplasmid system.

    Science.gov (United States)

    Loh, Qiuting; Leong, Siew Wen; Tye, Gee Jun; Choong, Yee Siew; Lim, Theam Soon

    2015-05-15

    The low presentation efficiency of Fab (fragment antigen binding) fragments during phage display is largely due to the complexity of disulphide bond formation. This can result in the presentation of Fab fragments devoid of a light chain during phage display. Here we propose the use of a coplasmid system encoding several molecular chaperones (DsbA, DsbC, FkpA, and SurA) to improve Fab packaging. A comparison was done using the Fab fragment from IgG and IgD. We found that the use of the coplasmid during phage packaging was able to improve the presentation efficiency of the Fab fragment on phage surfaces. A modified version of panning using the coplasmid system was evaluated and was successful at enriching Fab binders. Therefore, the coplasmid system would be an attractive alternative for improved Fab presentation for phage display. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Protein crystallization with microseed matrix screening: application to human germline antibody Fabs

    International Nuclear Information System (INIS)

    Obmolova, Galina; Malia, Thomas J.; Teplyakov, Alexey; Sweet, Raymond W.; Gilliland, Gary L.

    2014-01-01

    The power of microseed matrix screening is demonstrated in the crystallization of a panel of antibody Fab fragments. The crystallization of 16 human antibody Fab fragments constructed from all pairs of four different heavy chains and four different light chains was enabled by employing microseed matrix screening (MMS). In initial screening, diffraction-quality crystals were obtained for only three Fabs, while many Fabs produced hits that required optimization. Application of MMS, using the initial screens and/or refinement screens, resulted in diffraction-quality crystals of these Fabs. Five Fabs that failed to give hits in the initial screen were crystallized by cross-seeding MMS followed by MMS optimization. The crystallization protocols and strategies that resulted in structure determination of all 16 Fabs are presented. These results illustrate the power of MMS and provide a basis for developing future strategies for macromolecular crystallization

  4. Improved radioimaging and tumor localization with monoclonal F(ab')2

    International Nuclear Information System (INIS)

    Wahl, R.L.; Parker, C.W.; Philpott, G.W.

    1983-01-01

    Monoclonal anti-tumor antibodies have great promise for radioimmunodetection and localization of tumors. Fab and F(ab')2 fragments, which lack the Fc fragment of antibody (Ab), are cleared more rapidly from the circulation and may have less nonspecific tissue binding than intact Ab. In radioimaging studies using a murine monoclonal antibody to carcinoembryonic antigen in a human colon carcinoma xenografted into hamsters, F(ab')2 fragments were shown superior to Fab fragments and intact antibody for scintiscanning. In double-label experiments with anti-CEA antibody and control monoclonal IgG, F(ab')2 fragments were found to give better and more rapid specific tumor localization than intact antibody or Fab fragments. F(ab')2 fragments offer significant promise for tumor imaging and possibly therapy

  5. Opening the Arms: The FAB Projects and Digital Resilience

    Science.gov (United States)

    Longden, Alison; Monaghan, Tom; Mycroft, Lou; Kelly, Claire

    2018-01-01

    A chance observation in a staff training session led to a series of action research projects, which dug into why some educators are digitally resistant whilst others, however experienced, are not. The Education and Training Foundation-funded "FAB Projects" have so far spanned three years and three small-scale research investigations and…

  6. Scatterometry on pelliclized masks: an option for wafer fabs

    Science.gov (United States)

    Gallagher, Emily; Benson, Craig; Higuchi, Masaru; Okumoto, Yasuhiro; Kwon, Michael; Yedur, Sanjay; Li, Shifang; Lee, Sangbong; Tabet, Milad

    2007-03-01

    Optical scatterometry-based metrology is now widely used in wafer fabs for lithography, etch, and CMP applications. This acceptance of a new metrology method occurred despite the abundance of wellestablished CD-SEM and AFM methods. It was driven by the desire to make measurements faster and with a lower cost of ownership. Over the last year, scatterometry has also been introduced in advanced mask shops for mask measurements. Binary and phase shift masks have been successfully measured at all desired points during photomask production before the pellicle is mounted. There is a significant benefit to measuring masks with the pellicle in place. From the wafer fab's perspective, through-pellicle metrology would verify mask effects on the same features that are characterized on wafer. On-site mask verification would enable quality control and trouble-shooting without returning the mask to a mask house. Another potential application is monitoring changes to mask films once the mask has been delivered to the fab (haze, oxide growth, etc.). Similar opportunities apply to the mask metrologist receiving line returns from a wafer fab. The ability to make line-return measurements without risking defect introduction is clearly attractive. This paper will evaluate the feasibility of collecting scatterometry data on pelliclized masks. We explore the effects of several different pellicle types on scatterometry measurements made with broadband light in the range of 320-780 nm. The complexity introduced by the pellicles' optical behavior will be studied.

  7. Mechanism of immunoglobulin G4 Fab-arm exchange

    NARCIS (Netherlands)

    Rispens, Theo; Ooijevaar-de Heer, Pleuni; Bende, Onno; Aalberse, Rob C.

    2011-01-01

    Immunoglobulin G (IgG) antibodies are symmetrical molecules that may be regarded as covalent dimers of 2 half-molecules, each consisting of a light chain and a heavy chain. Human IgG4 is an unusually dynamic antibody, with half-molecule exchange ("Fab-arm exchange") resulting in asymmetrical,

  8. Bio-efficacy of gamma irradiation against pulse beetle, Callosobruchus maculatus L. infesting cowpea seeds

    International Nuclear Information System (INIS)

    Chauhan, Sumit Kumar; Bhalla, S.; Gautam, S.

    2015-01-01

    The pulse beetle, Callosobruchus maculatus is an important pest of cowpea in storage. It has cosmopolitan distribution with wide host range and also has different strains. It causes 20-60 per cent losses during storage of cowpea. Hazardous environmental effects posed by the chemicals used for its management necessitate the need for an alternative ecofriendly strategy to control the insect. Gamma irradiation seems to be a viable, non-chemical, residue-free, ecofriendly strategy. The experimental insect, C. maculatus was reared on the cowpea seeds under controlled conditions (28±1℃ and 65±5% RH). The freshly emerged adults (about 24-36 h of age) were exposed in very fine thin polythene envelops to gamma radiation at different doses viz., 25, 50, 100, 200, 400, 600 and 800 Gy using Cobalt-60 Gamma irradiation facility at Food Technology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, India. The parameters observed included adult mortality, longevity, fecundity of the survivors and adult emergence in F1 generation. Dose dependent insect mortality was observed with immediate mortality at higher doses. High mortality was observed within 24 hour of irradiation. However, complete mortality resulted within five days of irradiation at 600 and 800 Gy as compared to 12 days in control. Adult longevity decreased with increase in radiation dose. The mean longevity decreased from 6.00 days in control to 2.48 days at the highest dose. The eggs laid by the treated adult beetles did not develop into the adults of next generation. Much higher doses were required to kill the adult while the complete sterility (100% sterility) was found even at the lowest dose of 25 Gy. Thus, gamma irradiation has potential to be used as an eco-friendly mitigation measure against C. maculatus. (author)

  9. Insecticidal Activity and Chemical Composition of the Morinda lucida Essential Oil against Pulse Beetle Callosobruchus maculatus

    Directory of Open Access Journals (Sweden)

    Moses S. Owolabi

    2014-01-01

    Full Text Available Insecticidal activity of essential oil extracted from Morinda lucida was tested on pulse beetle Callosobruchus maculatus, which is a pest that causes serious damage to several pulses. The insecticidal activity was compared with two pesticides, Phostoxin and Primo-ban-20. 120 mixed sex adult C. maculatus were introduced, along with 30 g of cowpeas. Four concentrations (0.40, 0.20, 0.10, and 0.05 μg/mL of the M. lucida essential oil, Phostoxin, and Primo-ban-20 were tested. Essential oil chemical composition was analyzed by GC-MS. M. lucida essential oil showed a high toxicological effect, producing 100% mortality after 72 hours at a dose of 0.20 μg/mL. M. lucida essential oil had a potent insecticidal activity (LC90 = 0.629 μg/mL compared to both pesticides, Phostoxin (LC90 = 0.652 μg/mL and Primo-ban-20 (LC90 = 0.726 μg/mL, at 24 h. The main compounds of the essential oil were the oxygenated monoterpenoids, 1,8-cineole (43.4%, and α-terpinyl acetate (14.5%, and the monoterpene hydrocarbons, mostly sabinene (8.2% and β-pinene (4.0%. Results clearly indicate that M. lucida essential oil can be used as an effective alternative for pulse beetle C. maculatus control, and it could be tested against other pulse beetles affecting Asia and Africa and throughout the world, thereby reducing use of synthetic pesticides.

  10. Some gamma radiation effects on Callosobruchus maculatus (Fabr., 1775) (Col. Bruchidae)

    International Nuclear Information System (INIS)

    Walder, J.M.M.

    1975-10-01

    The effect of gamma irradiation from a Cobalt-60 source on Callosobruchus maculatus (Fabr.) was studied. Experiments showed that at a dose-rate of 9820 rad/hour the LD 75 of egg embryos was 2 Krad the LD 100 of larvae was 3 Krad and the LD 90 of pupae was 20 Krad. Total sterilization of adults irradiated of the pupae stage was 5 Krad. The sterilizing dose of irradiated adults was 8 Krad at a calculated dose-rate of 57.630 rad/hour

  11. Biological effects of some plant oils, gamma radiation and their interactive effects on callosobruchus maculatus (F.)

    International Nuclear Information System (INIS)

    EI-Orabi, M. N.; Shafei, D.M.Y.; Amer, A.M.

    2007-01-01

    Garlic, lemon and castor oils were evaluated for the control of infestation of Callosobruchus maculatus (F.). Oviposition, hatch ability, adult emergence were evaluated, Egg deposition was completely inhibited when cowpea seeds were treated with garlic, lemon and castor oils at doses 80,120 and 1300μl. Mean number of eggs laid by a female was significantly reduced by subjecting all developmental stages to increased radiation doses when the targeted sex was male, female or both. Egg deposition was. completely inhibited depending, on the irradiated sex and the developmental stage

  12. Surviving historical Patagonian landscapes and climate: molecular insights from Galaxias maculatus

    Directory of Open Access Journals (Sweden)

    Carrea Cecilia

    2010-03-01

    Full Text Available Abstract Background The dynamic geological and climatic histories of temperate South America have played important roles in shaping the contemporary distributions and genetic diversity of endemic freshwater species. We use mitochondria and nuclear sequence variation to investigate the consequences of mountain barriers and Quaternary glacial cycles for patterns of genetic diversity in the diadromous fish Galaxias maculatus in Patagonia (~300 individuals from 36 locations. Results Contemporary populations of G. maculatus, east and west of the Andes in Patagonia, represent a single monophyletic lineage comprising several well supported groups. Mantel tests using control region data revealed a strong positive relationship when geographic distance was modeled according to a scenario of marine dispersal. (r = 0.69, P = 0.055. By contrast, direct distance between regions was poorly correlated with genetic distance (r = -0.05, P = 0.463. Hierarchical AMOVAs using mtDNA revealed that pooling samples according to historical (pre-LGM oceanic drainage (Pacific vs. Atlantic explained approximately four times more variance than pooling them into present-day drainage (15.6% vs. 3.7%. Further post-hoc AMOVA tests revealed additional genetic structure between populations east and west of the Chilean Coastal Cordillera (coastal vs. interior. Overall female effective population size appears to have remained relatively constant until roughly 0.5 Ma when population size rapidly increased several orders of magnitude [100× (60×-190×] to reach contemporary levels. Maximum likelihood analysis of nuclear alleles revealed a poorly supported gene tree which was paraphyletic with respect to mitochondrial-defined haplogroups. Conclusions First diversifying in the central/north-west region of Patagonia, G. maculatus extended its range into Argentina via the southern coastal regions that join the Pacific and Atlantic oceans. More recent gene flow between northern

  13. Toxicity and repellence of African plants traditionally used for the protection of stored cowpea against Callosobruchus maculatus

    NARCIS (Netherlands)

    Boeke, S.J.; Baumgart, I.R.; Loon, van J.J.A.; Huis, van A.; Dicke, M.; Kossou, D.K.

    2004-01-01

    In a search for botanical products to control the main insect pest of stored cowpea, Callosobruchus maculatus, 33 traditionally used African plants were tested in the laboratory for their toxic and repellent effects against this beetle. Toxicity was evaluated measuring life history parameters in a

  14. Exposure to 4100K fluorescent light elicits sex specific transcriptional responses in Xiphophorus maculatus skin.

    Science.gov (United States)

    Boswell, William T; Boswell, Mikki; Walter, Dylan J; Navarro, Kaela L; Chang, Jordan; Lu, Yuan; Savage, Markita G; Shen, Jianjun; Walter, Ronald B

    2018-06-01

    It has been reported that exposure to artificial light may affect oxygen intake, heart rate, absorption of vitamins and minerals, and behavioral responses in humans. We have reported specific gene expression responses in the skin of Xiphophorus fish after exposure to ultraviolet light (UV), as well as, both broad spectrum and narrow waveband visible light. In regard to fluorescent light (FL), we have shown that male X. maculatus exposed to 4100K FL (i.e. "cool white") rapidly suppress transcription of many genes involved with DNA replication and repair, chromosomal segregation, and cell cycle progression in skin. We have also detailed sex specific transcriptional responses of Xiphophorus skin after exposure to UVB. However, investigation of gender differences in global gene expression response after exposure to 4100K FL has not been reported, despite common use of this FL source for residential, commercial, and animal facility illumination. Here, we compare RNA-Seq results analyzed to assess changes in the global transcription profiles of female and male X. maculatus skin in response to 4100K FL exposure. Our results suggest 4100K FL exposure incites a sex-biased genetic response including up-modulation of inflammation in females and down modulation of DNA repair/replication in males. In addition, we identify clusters of genes that become oppositely modulated in males and females after FL exposure that are principally involved in cell death and cell proliferation. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Negative Assortative Mating Based on Body Coloration in the Freshwater Platyfish (Poecillidae: Xiphophorus maculatus

    Directory of Open Access Journals (Sweden)

    Tyler E. Frankel

    2017-04-01

    Full Text Available The ability of individuals within a population to survive and thrive is highly dependent upon the maintenance of genetic variation and phenotypic diversity, thereby ensuring adaptation to dynamic environments. A fundamental method of maintaining such variation is through a negative assortative mating strategy, in which individuals would be expected to reproductively select members of the opposite sex that exhibit dissimilar phenotypes. Employing three uniform body color morphs, red, yellow and blue, of the platyfish (Xiphophorus maculatus, this study was designed to investigate whether X. maculatus females would preferentially be attracted to males exhibiting an alternative color, thereby enabling an examination of the effect of male body coloration on mate choice by adult females. Mate choice was determined based on the initial preference of each female, as well as the amount of time females spent associating with each male. Initial preferences were analyzed using a binomial distribution test, and overall preference data using Wilcoxon signed rank tests. Red females initially selected for dissimilar colored males, and spent a significantly larger amount of time associating with blue and yellow males, as did yellow females with red and blue males. Blue females initially selected and spent a significantly larger amount of time associating with red males but, interestingly, showed no selective preference between blue and yellow males. In these experimental trials, the overall strong mate selection exhibited by female platyfish for males of dissimilar coloration is suggestive of a negative assortative mating strategy and provides evidence for the maintenance of color polymorphism in nature populations.

  16. New morphological data on Cucullanus pinnai pinnai (Nematoda) parasitizing Pimelodus maculatus (Pimelodidae) in southeastern Brazil.

    Science.gov (United States)

    Vieira, Vivian Suane de Freitas; Vieira, Fabiano Matos; Luque, José Luis

    2015-01-01

    This paper describes the morphology of Cucullanus pinnai pinnai parasitizing Pimelodus maculatus in the Guandu River, Brazil, based on differential interference contrast (DIC) microscopy and scanning electron microscopy (SEM), providing new morphological data about this species of parasite. Nematodes were collected between May and October 2012 from specimens of Pimelodus maculatus in the Guandu River (22°48'2"S, 43°37'35"W), in the state of Rio de Janeiro, Brazil. Some characteristics of specimens of Cucullanus in this study fall within the range of morphological variations of previously studied C. pinnai pinnai. Most of the specimens studied here had excretory pore and deirids located at the posterior end of the oesophagus, a feature not recorded in previous studies of this species. In addition, the size of the gubernaculum was larger than the other specimens previously studied. The SEM and DIC analyses of C. pinnai revealed several morphological details of the cephalic region and the tail papillae. With regard to the polymorphism of C. pinnai, morphological and genetic studies of this cucullanid nematode are needed, involving large numbers of host species and a wide geographical distribution.

  17. Novel Infection Site and Ecology of Cryptic Didymocystis sp. (Trematoda) in the Fish Scomberomorus maculatus.

    Science.gov (United States)

    Schrandt, Meagan N; Andres, Michael J; Powers, Sean P; Overstreet, Robin M

    2016-06-01

    An undescribed, cryptic species of Didymocystis, as determined from sequences of 2 ribosomal genes and superficially similar to Didymocystis scomberomori ( MacCallum and MacCallum, 1916 ), infected the skin of the Spanish mackerel, Scomberomorus maculatus , in the north-central Gulf of Mexico (GOM). An analysis of 558 fish from 2011 to 2013 from Louisiana, Mississippi, Alabama, and the Florida panhandle showed the prevalence of the trematode varied both spatially and temporally but not with sex of the fish host. Month, year, and geographic location were identified by a negative binomial generalized linear model as indicators of the abundance and intensity of infection. Prevalence, abundance, and intensity of infection were greatest in spring and fall months off the Florida panhandle. Furthermore, the abundance and intensity of infection correlated negatively with fork length, weight, and gonad weight of mature fish but positively with longitude. Therefore, smaller adult fish tended to be more infected than larger adults, and prevalence and intensity increased from west to east (Louisiana to Florida). Spatial and temporal trends seemed to result from physical factors (e.g., water temperature, salinity, bottom type), but they also coincided with the annual migration of S. maculatus as fish moved northward along the GOM coastline from the southern tip of Florida in the spring months and returned in the fall, being present in the north-central GOM from late spring through fall. This pattern suggests the possibility that acquisition of infections occurred from a molluscan host in waters off the Florida panhandle.

  18. Haematological changes in Bufo maculatus treated with sublethal concentrations of Cadmium

    Directory of Open Access Journals (Sweden)

    Lawrence Ikechukwu Ezemonye

    2011-12-01

    Full Text Available Adult Bufo maculatus was exposed to sublethal cadmium concentrations of 0.25, 0.50, 1.00 and 2.00 mg/L. The toxicant from which the cadmium concentrations were prepared was cadmium chloride (CdCl2.H2O. There were three replicate tanks per treatment and three individuals per tank including control groups. The hematologic alterations based on the examination of blood indices during the 28 days of exposure showed that total erythrocyte count (TEC, hematocrit (Hct and hemoglobin (Hb concentration decreased (P<0.05 relative to controls. The decline was concentration- dependent as concentration of cadmium increased. The decline in hemoglobin and hematocrit in the experimental organism could be due to a decrease in the synthesis or release of erythrocytes into the circulation or an increase in the rate of erythrocyte destruction inflicted by cadmium toxicity. There was significant (P<0.05 elevation in total leuko- leukocyte count (TLC with increase in the concen- cyte concentration of cadmium. The increase in total leukocyte count observed in this study could be attributed to a stimulation of the immune system in response to tissue damage caused by cadmium toxicity. The study has shown that the exposure of the Bufo maculatus toad to cadmium can inflict alterations in the hematologic indices, which could induce unfavorable physiological changes in the amphibian, which may lead to death. There is, therefore, the need to protect amphibians in order to sustain the biodiversity in the Nigerian Niger Delta ecological zone.

  19. Effect of body size and temperature on respiration of Galaxias maculatus (Pisces: Galaxiidae)

    Science.gov (United States)

    Milano, D.; Vigliano, P.H.; Beauchamp, David A.

    2017-01-01

    Body mass and temperature are primary determinants of metabolic rate in ectothermic animals. Oxygen consumption of post-larval Galaxias maculatus was measured in respirometry trials under different temperatures (5–21°C) and varying body masses (0.1–>1.5 g) spanning a relevant range of thermal conditions and sizes. Specific respiration rates (R in g O2 g−1 d−1) declined as a power function of body mass and increased exponentially with temperature and was expressed as: R = 0.0007 * W −0.31 * e 0.13 * T. The ability of this model to predict specific respiration rate was evaluated by comparing observed values with those predicted by the model. Our findings suggest that the respiration rate of G. maculatus is the result of multiple interactive processes (intrinsic and extrinsic factors) that modulate each other in ‘meta-mechanistic’ ways; this would help to explain the species’ ability to undergo the complex ontogenetic habitat shifts observed in the lakes of the Andean Patagonic range.

  20. Introduction of a glycosylation site in the constant region decreases the aggregation of adalimumab Fab.

    Science.gov (United States)

    Nakamura, Hitomi; Oda-Ueda, Naoko; Ueda, Tadashi; Ohkuri, Takatoshi

    2018-06-18

    The production of therapeutic monoclonal antibodies is costly; therefore, antigen-binding fragments (Fabs) can be used instead. However, their tendency toward aggregation can reduce the half-life in the plasma and the therapeutic effectiveness. To examine the effect of glycosylation on the properties of the Fab of a therapeutic antibody, an N-glycosylation site was introduced at position 178 of the H-chain constant region of adalimumab Fab through site-directed mutagenesis of L178 N (H:L178 N Fab), and then H:L178 N Fab was expressed in Pichia pastoris. SDS-PAGE analysis with treatment of N-glycosidase F or periodic acid-Schiff reagent showed that H:L178 N Fab contained a relatively low glycan level. Moreover, the H:L178 N mutation did not decrease the binding activity and thermal stability of Fab, and H:L178 N Fab was more resistant to protease digestion than wild-type Fab. The aggregation of Fab induced by pH-shift stress was measured by monitoring the optical density at 350 nm. Although the wild-type Fab showed a large increase in optical density with an increase of protein concentration, no such increase of turbidity during aggregation was found in H:L178 N Fab. These results demonstrated that glycosylation at position 178 of the H-chain constant region of adalimumab Fab can prevent protein aggregation, and therefore serve as a potentially effective platform for drug development. Copyright © 2018. Published by Elsevier Inc.

  1. A single-domain antibody-linked Fab bispecific antibody Her2-S-Fab has potent cytotoxicity against Her2-expressing tumor cells.

    Science.gov (United States)

    Li, Aifen; Xing, Jieyu; Li, Li; Zhou, Changhua; Dong, Bin; He, Ping; Li, Qing; Wang, Zhong

    2016-12-01

    Her2, which is frequently overexpressed in breast cancer, is one of the most studied tumor-associated antigens for cancer therapy. Anti-HER2 monoclonal antibody, trastuzumab, has achieved significant clinical benefits in metastatic breast cancer. In this study, we describe a novel bispecific antibody Her2-S-Fab targeting Her2 by linking a single domain anti-CD16 VHH to the trastuzumab Fab. The Her2-S-Fab antibody can be efficiently expressed and purified from Escherichia coli, and drive potent cancer cell killing in HER2-overexpressing cancer cells. In xenograft model, the Her2-S-Fab suppresses tumor growth in the presence of human immune cells. Our results suggest that the bispecific Her2-S-Fab may provide a valid alternative to Her2 positive cancer therapy.

  2. Evaluation of strategies to control Fab light chain dimer during mammalian expression and purification: A universal one-step process for purification of correctly assembled Fab.

    Science.gov (United States)

    Spooner, Jennifer; Keen, Jenny; Nayyar, Kalpana; Birkett, Neil; Bond, Nicholas; Bannister, David; Tigue, Natalie; Higazi, Daniel; Kemp, Benjamin; Vaughan, Tristan; Kippen, Alistair; Buchanan, Andrew

    2015-07-01

    Fabs are an important class of antibody fragment as both research reagents and therapeutic agents. There are a plethora of methods described for their recombinant expression and purification. However, these do not address the issue of excessive light chain production that forms light chain dimers nor do they describe a universal purification strategy. Light chain dimer impurities and the absence of a universal Fab purification strategy present persistent challenges for biotechnology applications using Fabs, particularly around the need for bespoke purification strategies. This study describes methods to address light chain dimer formation during Fab expression and identifies a novel CH 1 affinity resin as a simple and efficient one-step purification for correctly assembled Fab. © 2015 Wiley Periodicals, Inc.

  3. Sampling methods for Graphoderus bilineatus (Coleoptera: Dytiscidae).

    NARCIS (Netherlands)

    Koese, B.; Cuppen, J.G.M.

    2006-01-01

    Onderzoek naar vangmethodes voor Graphoderus bilineatus (Coleoptera: Dytiscidae) Als onderdeel van een integrale studie naar de waterroofkever Graphoderus bilineatus in Nederland (in opdracht van het ministerie van lnv), werd een vergelijkend onderzoek uitgevoerd naar verschillende

  4. Immobilization and functional reconstitution of antibody Fab fragment by solid-phase refolding.

    Science.gov (United States)

    Kumada, Yoichi; Hamasaki, Kyoto; Nakagawa, Aya; Sasaki, Eiju; Shirai, Tatsunori; Okumura, Masahiro; Inoue, Manami; Kishimoto, Michimasa

    2013-12-31

    In this study, we demonstrated the successful preparation of a Fab antibody-immobilized hydrophilic polystyrene (phi-PS) plate via one- and two-step solid-phase refolding methods. Both polystyrene-binding peptide (PS-tag)-fused Fd fragment of heavy chain (Fab H-PS) and full-length of light-chain (Fab L-PS) were individually produced in insoluble fractions of Escherichia coli cells, and they were highly purified in the presence of 8M of urea. Antigen-binding activities of Fab antibody immobilized were correctly recovered by the one-step solid-phase refolding method that a mixture of Fab H-PS and Fab L-PS was immobilized in the presence of 0.5-2M urea, followed by surface washing of the phi-PS plate with PBST. These results indicate that by genetic fusion of a PS-tag, a complex between Fab H and Fab L was efficiently immobilized on the surface of a phi-PS plate even in the presence of a low concentration of urea, and was then correctly refolded to retain its high antigen-binding activity via removal of the urea. A two-step solid-phase refolding method whereby Fab H-PS and Fab L-PS were successively refolded on the surface of a phi-PS plate also resulted in Fab antibody formation on the plate. Furthermore, both the binding affinity and the specificity of the Fab antibody produced by the two-step method were highly maintained, according to the results of sandwich ELISA and competitive ELISA using Fab antibody-immobilized plate via two-step solid-phase refolding. Thus, the solid-phase refolding method demonstrated in this study should be quite useful for the preparation of a Fab antibody-immobilized PS surface with high efficiency from individually produced Fab H-PS and Fab L-PS. This method will be applicable to the preparation of a large Fab antibody library on the surface of a PS plate for use in antibody screening. © 2013. Published by Elsevier B.V. All rights reserved.

  5. A pilot pharmacokinetic study of tricyclic antidepressant ovine Fab for TCA poisoning in children.

    Science.gov (United States)

    Yalindağ-Oztürk, Nilüfer; Goto, Collin S; Shepherd, Greene; Torres, Olivia Nayeli; Giroir, Brett

    2010-06-01

    A pilot study of tricyclic antidepressant (TCA)-specific antibody fragments (TCA Fab) in TCA-intoxicated adults showed a marked increase in serum total TCA concentrations following TCA Fab infusion with no worsening signs of TCA toxicity. TCA Fab pharmacokinetics (PK) was not described in this adult study. The objective of this study was to evaluate the PK of TCA Fab in children with TCA poisoning. This was an open-label, single-center, dose escalation pilot trial of three patients. Inclusion criteria were documented TCA ingestion with at least one serious complication (QRS prolongation, dysrhythmia, hypotension, seizure, or coma). Patients were assigned to either a low-dose intravenous TCA Fab regimen (15, 30, and 60 mg/kg) or a high-dose regimen (30, 60, and 120 mg/kg) as needed to reverse TCA toxicity. Following the administration of TCA Fab, samples of blood and urine were obtained for PK evaluations. The outcomes of interest were serum and urine TCA concentrations (free and total), serum and urine Fab concentrations, improvement or worsening of TCA toxicity, and adverse effects. Three study patients were 11, 11, and 14 years of age. Two patients received 15 mg/kg of TCA Fab and one patient received a total of 90 mg/kg of TCA Fab (30 + 60 mg/kg). Serum-bound TCA increased significantly following TCA Fab administration with concomitant enhanced urinary elimination. Serum-free TCA concentrations were minimal to undetectable. Fab data were available for two patients. The serum TCA Fab area under the curve was 306.12 mg/L/h for the 15 mg/kg dose and 2,198.10 mg/L/h for the 90 mg/kg dose of TCA Fab. Maximum Fab concentrations correlated with maximum bound TCA in serum. The volume of distribution (V(D)) of TCA Fab was 0.2-0.3 L/kg. The clearance was 0.036-0.05 L/kg/h and the elimination half-life was 4 h. No adverse effects were observed. The limited PK data from this study are consistent with binding of TCA to TCA Fab and redistribution of TCA from the tissue to

  6. Wind Fab: Manufacturer of water pumping windmills in India

    International Nuclear Information System (INIS)

    Gurumoorthy, S.

    1991-01-01

    Wind Fab is one of the four manufacturers appointed recently by the Indian government to produce a classic gear type wind pump for deep well pumping (H = 50-100 meter). In various regions, the ground water table has been lowering considerably and a deep well wind pump is required. Wind Fab is still producing the 12PU500 windmill. The main problem is that the wind pumps are applied in a wide range of climatological, hydrological and agricultural conditions. Different types of windmills suitable for the different application ranges should be produced. It is explained that the drawings of the 12PU500 windmills were easily available and consequently the windmills were produced all over the country. There was no quality control and the 12PU500 was applied under low lift to high lift conditions. 5 figs., 5 tabs

  7. Spherical cows in the sky with fab four

    Energy Technology Data Exchange (ETDEWEB)

    Kaloper, Nemanja; Sandora, McCullen, E-mail: kaloper@physics.ucdavis.edu, E-mail: mesandora@ucdavis.edu [Department of Physics, University of California, Davis, CA 95616 (United States)

    2014-05-01

    We explore spherically symmetric static solutions in a subclass of unitary scalar-tensor theories of gravity, called the 'Fab Four' models. The weak field large distance solutions may be phenomenologically viable, but only if the Gauss-Bonnet term is negligible. Only in this limit will the Vainshtein mechanism work consistently. Further, classical constraints and unitarity bounds constrain the models quite tightly. Nevertheless, in the limits where the range of individual terms at large scales is respectively Kinetic Braiding, Horndeski, and Gauss-Bonnet, the horizon scale effects may occur while the theory satisfies Solar system constraints and, marginally, unitarity bounds. On the other hand, to bring the cutoff down to below a millimeter constrains all the couplings scales such that 'Fab Fours' can't be heard outside of the Solar system.

  8. A novel bispecific antibody, S-Fab, induces potent cancer cell killing.

    Science.gov (United States)

    Li, Li; He, Ping; Zhou, Changhua; Jing, Li; Dong, Bin; Chen, Siqi; Zhang, Ning; Liu, Yawei; Miao, Ji; Wang, Zhong; Li, Qing

    2015-01-01

    Bispecific antibodies that engage immune cells to kill cancer cells have been actively studied in cancer immunotherapy. In this study, we present a novel bispecific format, S-Fab, fabricated by linking a single-domain anti-carcinoembryonic antigen VHH to a conventional anti-CD3 Fab. In contrast to most bispecific antibodies, the S-Fab bispecific antibody can be efficiently expressed and purified from bacteria. The purified S-Fab is stable in serum and is able to recruit T cells to drive potent cancer cell killing. In xenograft models, the S-Fab antibody suppresses tumor growth in the presence of human immune cells. Our study suggested that the bispecific S-Fab format can be applied to a wide range of immunotherapies.

  9. Fab Four self-interaction in quantum regime

    Science.gov (United States)

    Arbuzov, A. B.; Latosh, B. N.

    2017-10-01

    Quantum behavior of the John Lagrangian from the Fab Four class of covariant Galileons is studied. We consider one-loop corrections to the John interaction due to cubic scalar field interaction. Counter terms are calculated, one appears because of massless scalar field theory infrared issues, another one lies in the George class, and the rest of them can be reduced to the initial Lagrangian up to surface terms. The role of quantum corrections in the context of cosmological applications is discussed.

  10. Fab Four self-interaction in quantum regime

    Energy Technology Data Exchange (ETDEWEB)

    Arbuzov, A.B.; Latosh, B.N. [JINR, Bogoliubov Laboratory for Theoretical Physics, Dubna (Russian Federation); Dubna State University, Dubna, Moscow Region (Russian Federation)

    2017-10-15

    Quantum behavior of the John Lagrangian from the Fab Four class of covariant Galileons is studied. We consider one-loop corrections to the John interaction due to cubic scalar field interaction. Counter terms are calculated, one appears because of massless scalar field theory infrared issues, another one lies in the George class, and the rest of them can be reduced to the initial Lagrangian up to surface terms. The role of quantum corrections in the context of cosmological applications is discussed. (orig.)

  11. Detection of experimental myocarditis by monoclonal antimyosin antibody, Fab fragment

    Energy Technology Data Exchange (ETDEWEB)

    Rezkalla, S.; Kloner, R.A.; Khaw, B.A.; Haber, E.; Fallon, J.T.; Smith, F.E.; Khatib, R.

    1989-02-01

    The purpose of this study was to determine whether monoclonal antimyosin Fab (antigen binding fragment) was capable of labeling hearts with experimental coxsackievirus myocarditis, and to determine whether Fab could be used for detecting myocardial damage in either early or chronic phases of the disease. Sixty-five, 3-week-old cesarean-derived 1 (CD 1) mice were divided into two groups: group I (noninfected animals) and group II (infected with coxsackievirus B3). Mice from each group were killed on days 7, 17, 30, or 90 of infection. Forty-eight hours before killing, mice were injected with monoclonal I-125 antimyosin, Fab (25 microCi/injection) and radioactivity was counted in the heart. Selected heart sections were also examined by autoradiography. Heart radioactivity, count/m/mg (m +/- SEM) on days 7, 17, 30, and 90 of infection was 10.8 +/- 1.7, 21.3 +/- 1.1, 11.2 +/- 3.4, and 12.4 +/- 1.5 for group I, versus 36.7 +/- 8.0 (p less than 0.01), 50.0 +/- 4.5 (p less than 0.001), 33.4 +/- 16.1 (p = NS), and 40.6 +/- 8.5 (p less than 0.01) for group II, respectively. Autoradiography revealed focal uptake within areas of necrotic myocardium. We conclude that I125 Fab may be useful in detecting myocardial damage in the experimental model of murine myocarditis up to day 90 of infection.

  12. Detection of experimental myocarditis by monoclonal antimyosin antibody, Fab fragment

    International Nuclear Information System (INIS)

    Rezkalla, S.; Kloner, R.A.; Khaw, B.A.; Haber, E.; Fallon, J.T.; Smith, F.E.; Khatib, R.

    1989-01-01

    The purpose of this study was to determine whether monoclonal antimyosin Fab (antigen binding fragment) was capable of labeling hearts with experimental coxsackievirus myocarditis, and to determine whether Fab could be used for detecting myocardial damage in either early or chronic phases of the disease. Sixty-five, 3-week-old cesarean-derived 1 (CD 1) mice were divided into two groups: group I (noninfected animals) and group II (infected with coxsackievirus B3). Mice from each group were killed on days 7, 17, 30, or 90 of infection. Forty-eight hours before killing, mice were injected with monoclonal I-125 antimyosin, Fab (25 microCi/injection) and radioactivity was counted in the heart. Selected heart sections were also examined by autoradiography. Heart radioactivity, count/m/mg (m +/- SEM) on days 7, 17, 30, and 90 of infection was 10.8 +/- 1.7, 21.3 +/- 1.1, 11.2 +/- 3.4, and 12.4 +/- 1.5 for group I, versus 36.7 +/- 8.0 (p less than 0.01), 50.0 +/- 4.5 (p less than 0.001), 33.4 +/- 16.1 (p = NS), and 40.6 +/- 8.5 (p less than 0.01) for group II, respectively. Autoradiography revealed focal uptake within areas of necrotic myocardium. We conclude that I125 Fab may be useful in detecting myocardial damage in the experimental model of murine myocarditis up to day 90 of infection

  13. Rasch Analysis of the Fullerton Advanced Balance (FAB) Scale

    Science.gov (United States)

    Fiedler, Roger C.; Rose, Debra J.

    2011-01-01

    ABSTRACT Purpose: This cross-sectional study explores the psychometric properties and dimensionality of the Fullerton Advanced Balance (FAB) Scale, a multi-item balance test for higher-functioning older adults. Methods: Participants (n=480) were community-dwelling adults able to ambulate independently. Data gathering consisted of survey and balance performance assessment. Psychometric properties were assessed using Rasch analysis. Results: Mean age of participants was 76.4 (SD=7.1) years. Mean FAB Scale scores were 24.7/40 (SD=7.5). Analyses for scale dimensionality showed that 9 of the 10 items fit a unidimensional measure of balance. Item 10 (Reactive Postural Control) did not fit the model. The reliability of the scale to separate persons was 0.81 out of 1.00; the reliability of the scale to separate items in terms of their difficulty was 0.99 out of 1.00. Cronbach's alpha for a 10-item model was 0.805. Items of differing difficulties formed a useful ordinal hierarchy for scaling patterns of expected balance ability scoring for a normative population. Conclusion: The FAB Scale appears to be a reliable and valid tool to assess balance function in higher-functioning older adults. The test was found to discriminate among participants of varying balance abilities. Further exploration of concurrent validity of Rasch-generated expected item scoring patterns should be undertaken to determine the test's diagnostic and prescriptive utility. PMID:22210989

  14. Rasch Analysis of the Fullerton Advanced Balance (FAB) Scale.

    Science.gov (United States)

    Klein, Penelope J; Fiedler, Roger C; Rose, Debra J

    2011-01-01

    This cross-sectional study explores the psychometric properties and dimensionality of the Fullerton Advanced Balance (FAB) Scale, a multi-item balance test for higher-functioning older adults. Participants (n=480) were community-dwelling adults able to ambulate independently. Data gathering consisted of survey and balance performance assessment. Psychometric properties were assessed using Rasch analysis. Mean age of participants was 76.4 (SD=7.1) years. Mean FAB Scale scores were 24.7/40 (SD=7.5). Analyses for scale dimensionality showed that 9 of the 10 items fit a unidimensional measure of balance. Item 10 (Reactive Postural Control) did not fit the model. The reliability of the scale to separate persons was 0.81 out of 1.00; the reliability of the scale to separate items in terms of their difficulty was 0.99 out of 1.00. Cronbach's alpha for a 10-item model was 0.805. Items of differing difficulties formed a useful ordinal hierarchy for scaling patterns of expected balance ability scoring for a normative population. The FAB Scale appears to be a reliable and valid tool to assess balance function in higher-functioning older adults. The test was found to discriminate among participants of varying balance abilities. Further exploration of concurrent validity of Rasch-generated expected item scoring patterns should be undertaken to determine the test's diagnostic and prescriptive utility.

  15. A novel engineered interchain disulfide bond in the constant region enhances the thermostability of adalimumab Fab.

    Science.gov (United States)

    Nakamura, Hitomi; Oda-Ueda, Naoko; Ueda, Tadashi; Ohkuri, Takatoshi

    2018-01-01

    We constructed a system for expressing the Fab of the therapeutic human monoclonal antibody adalimumab at a yield of 20 mg/L in the methylotrophic yeast Pichia pastoris. To examine the contribution of interchain disulfide bonds to conformational stability, we prepared adalimumab Fab from which the interchain disulfide bond at the C-terminal region at both the CH 1 and CL domains was deleted by substitution of Cys with Ala (Fab ΔSS ). DSC measurements showed that the Tm values of Fab ΔSS were approximately 5 °C lower than those of wild-type Fab, suggesting that the interchain disulfide bond contributes to conformational thermostability. Using computer simulations, we designed a novel interchain disulfide bond outside the C-terminal region to increase the stability of Fab ΔSS . The resulting Fab (mutSS Fab ΔSS ) had the mutations H:V177C and L:Q160C in Fab ΔSS , confirming the formation of the disulfide bond between CH 1 and CL. The thermostability of mutSS Fab ΔSS was approximately 5 °C higher than that of Fab ΔSS . Therefore, the introduction of the designed interchain disulfide bond enhanced the thermostability of Fab ΔSS and mitigated the destabilization caused by partial reduction of the interchain disulfide bond at the C-terminal region, which occurs in site-specific modification such as PEGylation. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. The reliability and validity of the Turkish version of Fullerton Advanced Balance (FAB-T) scale.

    Science.gov (United States)

    Iyigun, Gozde; Kirmizigil, Berkiye; Angin, Ender; Oksuz, Sevim; Can, Filiz; Eker, Levent; Rose, Debra J

    2018-06-04

    The aim of this study was to evaluate the reliability and validity of the Turkish version of the FAB(FAB-T) scale in the older Turkish adults. The reliability and validity of the scale was tested on 200 community-dwelling older adults. FAB-T scale was scored by different physiotherapists on different days to evaluate inter-rater and intrarater reliability. The Berg Balance Scale (BBS) was used for the evaluation of convergent validity, and the content validity of the FAB-T scale was investigated. The FAB-T scale showed very high inter- and intra-rater reliability. For inter-rater agreement, on the individual test items and total score ICC values were 0.92 (95 %CI; 0.90-0.94) and 0.96 (95% CI; 0.95-0.97) respectively. The intra-rater agreement, on the individual test items and total score ICC values were 0.93 (95 %CI; 0.91- 0.95) and 0.96 (95% CI; 0.95- 0.97) respectively. There was a good agreement between the FAB-T and BBS scales. A high correlation was found between the BBS and FAB-T scales [rho = 0.70 (%95 CI; 0.62-0.76)] indicating good convergent validity. Considering the content validity of the FAB-T scale, no floor (floor score: 0%) or ceiling (ceiling score: 6.5%) effect was detected. The FAB-T scale was successfully translated from the original English version (FAB) and demonstrated strong psychometric features. It was found that the FAB-T scale has very high inter-rater and intra-rater reliability. Considering the convergent validity, the scale has high correlation with the BBS. The FAB-T has no floor and ceiling effect. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. Complex Binding of the FabR Repressor of Bacterial Unsaturated Fatty Acid Biosynthesis to its Cognate Promoters

    OpenAIRE

    Feng, Youjun; Cronan, John E.

    2011-01-01

    Two transcriptional regulators, the FadR activator and the FabR repressor control biosynthesis of unsaturated fatty acids in Escherichia coli. FabR represses expression of the two genes, fabA and fabB, required for unsaturated fatty acid synthesis and has been reported to require the presence of an unsaturated thioester (of either acyl carrier protein or CoA) in order to bind the fabA and fabB promoters in vitro. We report in vivo experiments in which unsaturated fatty acid synthesis was bloc...

  18. Assessment of Digoxin-Specific Fab Fragment Dosages in Digoxin Poisoning.

    Science.gov (United States)

    Nordt, Sean Patrick; Clark, Richard F; Machado, Carol; Cantrell, F Lee

    2016-01-01

    Digoxin poisoning still remains a common cause of morbidity and mortality. Fortunately, digoxin-specific Fab fragments are commercially available as an antidote. However, these Fab fragments are several thousand dollars per vial. There is a standardized formula to calculate appropriate Fab fragment dosage based on the serum digoxin concentration. This can greatly reduce the amount of Fab fragment administered. There is also an empiric dosing guideline recommending 6-10 vials be given; however, this may result in higher amounts of Fab fragments being administered than required. We performed this study to assess the amounts of digoxin-specific Fab fragments administered in the treatment of digoxin poisonings recorded in a poison control system database from January 1, 2000, to December 31, 2009, in which digoxin serum concentrations were available. This was a retrospective study of 278 patients, 107 with acute poisonings (group A) and 171 following chronic poisoning (group B). In group A, the calculated Fab dose was higher than the calculated dose based on available concentrations in 39 (36%) of group A and 15 (9%) of group B patients. The average wholesale price cost of the excessive dosages ranged from $4818 to as high as $50,589 per patient. Our data suggests that clinician education on digoxin poisoning and the use of the standardized formula to calculate the Fab dose may decrease over utilization and decrease costs associated with the administration of digoxin-specific Fab fragments in the treatment of digoxin poisonings.

  19. Frontal assessment battery (FAB) performance following traumatic brain injury hospitalized in an acute care setting.

    Science.gov (United States)

    Rojas, Natalia; Laguë-Beauvais, Maude; Belisle, Arielle; Lamoureux, Julie; AlSideiri, Ghusn; Marcoux, Judith; Maleki, Mohammed; Alturki, Abdulrahman Y; Anchouche, Sonia; Alquraini, Hanan; Feyz, Mitra; Guise, Elaine de

    2018-01-19

    The Frontal Assessment Battery (FAB) has been shown to be useful in several clinical settings. The aim of the present study was to examine the performance of patients with traumatic brain injury (TBI) on the FAB and to predict their acute outcome. The FAB was administered to 89 patients with mild (27 = uncomplicated and 39 = complicated) and moderate (n = 23) TBI during hospitalization in an acute care setting. The length of stay in days (LOS), Glasgow Outcome Scale-Revised score (GOSE) and Disability Rating Scale (DRS) score were collected. Results showed no significant differences between the three groups on the FAB score, but age and education were significantly associated with the FAB score. Parietal lesions were associated with lower total FAB score, and with the Similarities, Motor series and Conflicting instructions subscales, while frontal lesions were associated with lower performance on the Motor series and Conflicting instructions subscales. Total FAB score was significantly correlated with all outcome measures, and together the FAB total score and the Glasgow Coma Scale (GCS) score explained 30.8% of the variance in the DRS score. The FAB may be useful clinically to acutely assess frontal and parietal lobe functions at bedside in patients with TBI and, in combination with the GCS score to measure TBI severity, can enable clinicians to predict early outcome.

  20. Efficient production of Trastuzumab Fab antibody fragments in Brevibacillus choshinensis expression system.

    Science.gov (United States)

    Mizukami, Makoto; Onishi, Hiromasa; Hanagata, Hiroshi; Miyauchi, Akira; Ito, Yuji; Tokunaga, Hiroko; Ishibashi, Matsujiro; Arakawa, Tsutomu; Tokunaga, Masao

    2018-10-01

    The Brevibacillus expression system has been successfully employed for the efficient productions of a variety of recombinant proteins, including enzymes, cytokines, antigens and antibody fragments. Here, we succeeded in secretory expression of Trastuzumab Fab antibody fragments using B. choshinensis/BIC (Brevibacillus in vivocloning) expression system. In the fed-batch high-density cell culture, recombinant Trastuzumab Fab with amino-terminal His-tag (His-BcFab) was secreted at high level, 1.25 g/liter, and Fab without His-tag (BcFab) at ∼145 mg/L of culture supernatant. His-BcFab and BcFab were purified to homogeneity using combination of conventional column chromatographies with a yield of 10-13%. This BcFab preparation exhibited native structure and functions evaluated by enzyme-linked immunosorbent assay, surface plasmon resonance, circular dichroism measurements and size exclusion chromatography. To our knowledge, this is the highest production of Fab antibody fragments in gram-positive bacterial expression/secretion systems. Copyright © 2018 Elsevier Inc. All rights reserved.

  1. Aerial imaging for FABs: productivity and yield aspects

    Science.gov (United States)

    Englard, Ilan; Cohen, Yaron; Elblinger, Yair; Attal, Shay; Berns, Neil; Shoval, Lior; Ben-Yishai, Michael; Mangan, Shmoolik

    2009-03-01

    The economy of wafer fabs is changing faster for 3x geometry requirements and below. Mask set and exposure tool costs are almost certain to increase the overall cost per die requiring manufacturers to develop productivity and yield improvements to defray the lithography cell economic burden. Lithography cell cost effectiveness can be significantly improved by increasing mask availability while reducing the amount of mask sets needed during a product life cycle. Further efficiency can be gained from reducing send-ahead wafers and qualification cycle time, and elimination of inefficient metrology. Yield is the overriding die cost modulator and is significantly more sensitive to lithography as a result of masking steps required to fabricate the integrated circuit. Thus, for productivity to increase with minimal yield risk, the sample space of reticle induced source of variations should be large, with shortest measurement acquisition time possible. This paper presents the latest introduction of mask aerial imaging technology for the fab, Aera2TM for Lithography with IntenCTM, as an enabler for efficient lithography manufacturing. IntenCD is a high throughput, high density mask-based critical dimension (CD) mapping technology, with the potential for increasing productivity and yield in a wafer production environment. Connecting IntenCD to a feed forward advance process control (APC) reduces significantly the amount of traditional CD metrology required for robust wafer CD uniformity (CDU) correction and increases wafer CD uniformity. This in turn improves the lithography process window and yield and contributes to cost reduction and cycle time reduction of new reticles qualification. Advanced mask technology has introduced a new challenge. Exposure to 193nm wavelength stimulates haze growth on the mask and imposes a regular cleaning schedule. Cleaning eventually causes mask degradation. Haze growth impacts mask CD uniformity and induce global transmission fingerprint

  2. Revision of the Neotropical diving beetle genus Hydrodessus J. Balfour-Browne, 1953 (Coleoptera, Dytiscidae, Hydroporinae, Bidessini).

    Science.gov (United States)

    Miller, Kelly B

    2016-01-01

    The Neotropical diving beetle genus Hydrodessus J. Balfour-Browne, 1953 (Coleoptera: Dytiscidae: Hydroporinae: Bidessini) is revised. Thirty species are recognized. The following new species are described: Hydrodessus bimaculatus sp. n. (Venezuela), Hydrodessus brevis sp. n. (Venezuela), Hydrodessus concolorans sp. n. (Venezuela), Hydrodessus continuus sp. n. (Venezuela), Hydrodessus disjunctus sp. n. (Suriname), Hydrodessus fasciatus sp. n. (Brazil), Hydrodessus imparilis sp. n. (Ecuador), Hydrodessus keithi sp. n. (Brazil, Colombia, Ecuador), Hydrodessus kurti sp. n. (Suriname), Hydrodessus kylei sp. n. (Suriname, Venezuela), Hydrodessus laetus sp. n. (Venezuela), Hydrodessus latotibialis sp. n. (Peru), Hydrodessus maculatus sp. n. (Guyana, Venezuela), Hydrodessus morsus sp. n. (Venezuela), Hydrodessus palus sp. n. (Venezuela), and Hydrodessus tenuatus sp. n. (Suriname). The following new synonyms are established: Hydrodessus fragrans Spangler, 1985 = Hydrodessus biguttatus (Guignot, 1957) syn. n. and Hydrodessus robinae Spangler, 1985 = Hydrodessus octospilus (Guignot, 1957), syn. n. One species is transferred from Hydrodessus to Amarodytes Régimbart, Amarodytes soekhnandanae (Makhan, 1994), comb. n. Habitus photographs (dorsal and lateral) and photos of the ventral surfaces are provided for most species. Line drawings of male and female genitalia and other diagnostic features are also provided along with distribution maps.

  3. Acute hypersensitivity reaction to Crotalidae polyvalent immune Fab (CroFab) as initial presentation of galactose-α-1,3-galactose (α-gal) allergy.

    Science.gov (United States)

    Rizer, Justin; Brill, Kaitlin; Charlton, Nathan; King, Joshua

    2017-08-01

    Crotalidae polyvalent immune Fab antivenom (CroFab), commonly used for the treatment of clinically significant North American crotalinae envenomation, is generally well-tolerated. A novel form of anaphylaxis due to an IgE antibody response to the mammalian oligosaccharide galactose-α-1,3-galactose (α-gal) has been established following red-meat consumption as well as IV administration of cetuximab, which contain the α-gal epitope. We present a case of α-gal allergy discovered after acute hypersensitivity reaction to FabAV. A 61-year-old healthy female was bitten on her left ankle by Agkistrodon contortrix. Given the patient's rapid progression of pain and swelling, she was given FabAV. During infusion of FabAV, she developed diffuse hives over her entire body and itching, but denied respiratory or gastrointestinal symptoms and her vital signs remained stable. The FabAV was immediately discontinued and she received intravenous diphenhydramine and famotidine with gradual resolution of symptoms. On further discussion, she denied a history of α-gal or papaya allergy but rarely ate red meat and endorsed sustaining frequent tick bites. Subsequent antibody testing was significant for an α-1,3-galactose IgE concentration of 45,000 U/L (normal allergy. To our knowledge, this is the first report of FabAV hypersensitivity associated with an underlying α-gal allergy.

  4. IgG Fab Fragments Forming Bivalent Complexes by a Conformational Mechanism That Is Reversible by Osmolytes*

    Science.gov (United States)

    Nelson, Alfreda D.; Hoffmann, Michele M.; Parks, Christopher A.; Dasari, Surendra; Schrum, Adam G.; Gil, Diana

    2012-01-01

    Generated by proteolytic cleavage of immunoglobulin, Fab fragments possess great promise as blocking reagents, able to bind receptors or other targets without inducing cross-linking. However, aggregation of Fab preparations is a common occurrence, which generates intrinsic stimulatory capacity and thwarts signal blockade strategies. Using a panel of biochemical approaches, including size exclusion chromatography, SDS-PAGE, mass spectrometry, and cell stimulation followed by flow cytometry, we have measured the oligomerization and acquisition of stimulatory capacity that occurs in four monoclonal IgG Fabs specific for TCR/CD3. Unexpectedly, we observed that all Fabs spontaneously formed complexes that were precisely bivalent, and these bivalent complexes possessed most of the stimulatory activity of each Fab preparation. Fabs composing bivalent complexes were more susceptible to proteolysis than monovalent Fabs, indicating a difference in conformation between the Fabs involved in these two different states of valency. Because osmolytes represent a class of compounds that stabilize protein folding and conformation, we sought to determine the extent to which the amino acid osmolyte l-proline might impact bivalent Fab complexation. We found that l-proline (i) inhibited the adoption of the conformation associated with bivalent complexation, (ii) preserved Fab monovalency, (iii) reversed the conformation of preformed bivalent Fabs to that of monovalent Fabs, and (iv) separated a significant percentage of preformed bivalent complexes into monovalent species. Thus, Fab fragments can adopt a conformation that is compatible with folding or packing of a bivalent complex in a process that can be inhibited by osmolytes. PMID:23109335

  5. Quarantine treatment by gamma radiation for different development stages of Callosobruchus maculatus in bean Vigna sinensis

    International Nuclear Information System (INIS)

    Arthur, Valter; Machi, André R.; Franco, Suely S.H.

    2017-01-01

    The loss of stored grain caused by insects generates a problem of economic order of importance, due to concern about the increased supply of food for the world population is expanding. Associated with this fact, there is the problem of nutritional deficiency due to lack of protein, especially for the less privileged populations. The use of ionizing radiation in grains and products stored without a doubt can solve the problem of the losses in these products, since it does not induce resistance to insects and leaves no toxic residue in the products, and is considered an effective and safe method. The aim of the experiment was to determine the effect of ionizing radiation from cobalt-60 as a quarantine treatment for the different stages of development of Callosobruchus maculatus (Fabr., 1972) (Coleóptera, Chysomilidae) in bean Vigna sinensis. The experiment was conducted in the laboratory of Radiobiology and Environment CENA/USP, Piracicaba, SP, Brazil. Bean samples infested with eggs, larvae, pre-pupae and pupae C. maculatus, the experiment consisted of 4 replicates for each stage of the insect's life cycle, and each repetition consisted of 20 individuals (eggs, larvae, pre-pupae and pupae), a total of 200 subjects per treatment which were irradiated with doses of 0 (control), 25, 50, 75 and 100 Gy, a source of cobalt-60, Gammabeam-650 type, in a rate dose of 1.3 kGy / h. The experiment was conducted in a room with a relative of 25 ± 2 ° C temperature and humidity of 70 ± 5%. After 35 days of irradiation process were carried out evaluations of the number of insects emerged in each repetition within the treatments. From the results obtained it was concluded that the dose lethal to eggs and larvae was 25 Gy, while for pre-pupae was 50 Gy, to pupae 100 Gy was not sufficient to control the adult emergence. (author)

  6. Competitiveness values of irradiated adults of callosobruchus maculatus (F.) irradiated as mature pupae

    International Nuclear Information System (INIS)

    Ahmed, M.Y.Y.

    1981-01-01

    When mature pupae of Callosobruchus maculatus were treated with 3 Krad, the resulting adults were sterile when they were paired with untreated opposite sex. Males and females both treated with a sterilizing dose (3 Krad) and confined with untreated (U) males and females at a I male: I female: U male: U female (irradiated males: irradiated females: unirradiated males: unirradiated females) ratio caused 69.1% infertility in the resulting eggs. When the ratio of sterile males and females was increased to 5.5:1:1; 10:10:1:1 or 15:1:1 (I male: female: U male: U female) the percentage infertility reached 82.5, 95.0 and 100.0, respectively. The percentage of observed infertility was less than the expected infertility for the ratios 1:1:1:1:5:5:1:1 and 10:10:1:1, but it was exceeded with the highest ratio used (15:15:1:1). Competitiveness values for irradiated adults increased with an increasing ratio of irradiated to unirradiated adults. Since the ratio of 15:15:1:1 gave rise to 100% egg infertility (the expected infertility was 99.6%), no F 1 adults was produced; and the competitiveness value slightly exceeded 1.0 (i.e. the sterile adults were fully competitive with the normal ones). These results indicated that irradiation with 3 Krad, a sterilizing dose, did not decrease sexual competitiveness of irradiated adults. Also, the release of (I) females together with (I) males could give good results in controlling a population of C. maculatus in a autocidal control program; and, therefore, separation of the sexes prior to release is probably unnecessary. (author)

  7. Quarantine treatment by gamma radiation for different development stages of Callosobruchus maculatus in bean Vigna sinensis

    Energy Technology Data Exchange (ETDEWEB)

    Arthur, Valter; Machi, André R.; Franco, Suely S.H., E-mail: arthur@cena.usp.br [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). laboratório de Radiobiologia e Ambiente; Fontes, Lucia da Silva, E-mail: lsfontes@uol.com.br [Universidade Federal do Piauí (UFPI), Teresina, PI (Brazil). Dept. de Biologia; Harder, Márcia N.C., E-mail: marcia.harder@fatec.sp.gov.br [Faculdade de Tecnologia de Piracicaba (FATEC), SP (Brazil). Dep. Roque Trevisan; Rossi, Rodrigo S.; Arthur, Paula B.; Franco, José G., E-mail: paula.arthur@hotmail.com, E-mail: zegilmar60@gamil.com, E-mail: gilmita@uol.com.br [Instituto de Pesquisas Energéticas e Nucleares (IPEN/CNEN-SP), São Paulo, SP (Brazil)

    2017-07-01

    The loss of stored grain caused by insects generates a problem of economic order of importance, due to concern about the increased supply of food for the world population is expanding. Associated with this fact, there is the problem of nutritional deficiency due to lack of protein, especially for the less privileged populations. The use of ionizing radiation in grains and products stored without a doubt can solve the problem of the losses in these products, since it does not induce resistance to insects and leaves no toxic residue in the products, and is considered an effective and safe method. The aim of the experiment was to determine the effect of ionizing radiation from cobalt-60 as a quarantine treatment for the different stages of development of Callosobruchus maculatus (Fabr., 1972) (Coleóptera, Chysomilidae) in bean Vigna sinensis. The experiment was conducted in the laboratory of Radiobiology and Environment CENA/USP, Piracicaba, SP, Brazil. Bean samples infested with eggs, larvae, pre-pupae and pupae C. maculatus, the experiment consisted of 4 replicates for each stage of the insect's life cycle, and each repetition consisted of 20 individuals (eggs, larvae, pre-pupae and pupae), a total of 200 subjects per treatment which were irradiated with doses of 0 (control), 25, 50, 75 and 100 Gy, a source of cobalt-60, Gammabeam-650 type, in a rate dose of 1.3 kGy / h. The experiment was conducted in a room with a relative of 25 ± 2 ° C temperature and humidity of 70 ± 5%. After 35 days of irradiation process were carried out evaluations of the number of insects emerged in each repetition within the treatments. From the results obtained it was concluded that the dose lethal to eggs and larvae was 25 Gy, while for pre-pupae was 50 Gy, to pupae 100 Gy was not sufficient to control the adult emergence. (author)

  8. Neutron Reflection Study of Surface Adsorption of Fc, Fab, and the Whole mAb.

    Science.gov (United States)

    Li, Zongyi; Li, Ruiheng; Smith, Charles; Pan, Fang; Campana, Mario; Webster, John R P; van der Walle, Christopher F; Uddin, Shahid; Bishop, Steve M; Narwal, Rojaramani; Warwicker, Jim; Lu, Jian Ren

    2017-07-12

    Characterizing the influence of fragment crystallization (Fc) and antigen-binding fragment (Fab) on monoclonal antibody (mAb) adsorption at the air/water interface is an important step to understanding liquid mAb drug product stability during manufacture, shipping, and storage. Here, neutron reflection is used to study the air/water adsorption of a mAb and its Fc and Fab fragments. By varying the isotopic contrast, the adsorbed amount, thickness, orientation, and immersion of the adsorbed layers could be determined unambiguously. While Fc adsorption reached saturation within the hour, its surface adsorbed amount showed little variation with bulk concentration. In contrast, Fab adsorption was slower and the adsorbed amount was concentration dependent. The much higher Fc adsorption, as compared to Fab, was linked to its lower surface charge. Time and concentration dependence of mAb adsorption was dominated by Fab behavior, although both Fab and Fc behaviors contributed to the amount of mAb adsorbed. Changing the pH from 5.5 to 8.8 did not much perturb the adsorbed amount of Fc, Fab, or mAb. However, a small decrease in adsorption was observed for the Fc over pH 8-8.8 and vice versa for the Fab and mAb, consistent with a dominant Fab behavior. As bulk concentration increased from 5 to 50 ppm, the thicknesses of the Fc layers were almost constant at 40 Å, while Fab and mAb layers increased from 45 to 50 Å. These results imply that the adsorbed mAb, Fc, and Fab all retained their globular structures and were oriented with their short axial lengths perpendicular to the interface.

  9. A Study of the Charge Trap Transistor (CTT) for Post-Fab Modification of Wafers

    Science.gov (United States)

    2018-04-01

    AFRL-RY-WP-TR-2018-0030 A STUDY OF THE CHARGE TRAP TRANSISTOR (CTT) FOR POST- FAB MODIFICATION OF WAFERS Subramanian S. Iyer University of California...Final 13 June 2016 – 13 December 2017 4. TITLE AND SUBTITLE A STUDY OF THE CHARGE TRAP TRANSISTOR (CTT) FOR POST- FAB MODIFICATION OF WAFERS 5a. CONTRACT

  10. Phage-display libraries of murine and human antibody Fab fragments

    DEFF Research Database (Denmark)

    Engberg, J; Andersen, P S; Nielsen, L K

    1996-01-01

    We provide efficient and detailed procedures for construction, expression, and screening of comprehensive libraries of murine or human antibody Fab fragments displayed on the surface of filamentous phage. In addition, protocols for producing and using ultra-electrocompetent cells, for producing Fab...

  11. Use of isothiocyanatobenzyl-DTPA derivatized monoclonal antimyosin fab for enhanced in vivo target localization

    International Nuclear Information System (INIS)

    Khaw, B.A.; Gansow, O.; Brechbiel, M.W.; O'Donnell, S.M.; Nossiff, N.

    1990-01-01

    Monoclonal antimyosin Fab (AM-Fab) was radiolabeled with 111In via a new bifunctional chelating agent, isothiocyanatobenzyl-DTPA (SCN-DTPA), and used to visualize acute reperfused experimental myocardial infarction. Antibody localization was compared to 201 Tl (0.6 mCi) distribution in nine animals. Each animal was injected intravenously with 0.5 mCi of 111 In-SCN-DTPA AM-Fab preparations (Prep 1 [n = 5] and 2 [n = 4]). The biodistribution was compared to that of 111 In-labeled conventional bicyclic anhydride DTPA-AM-Fab (n = 5). 111 In-SCN-DTPA AM-Fab Prep 1 (lowest specific activity) showed highest specific target localization (31.6 +/- 3.5, MEAN infarct[0-20% Tl-201] to normal ration +/- SE) and lowest hepatic sequestration (0.0108 +/- 0.002% ID/g). Prep 2 showed similar infarct localization (18.4 +/- 1.2) to control 111 In-DTPA AM-Fab (16.9 +/- 1.1), but had higher hepatic activity (0.0326 +/- 0.014 and 0.0267 +/- 0.006 respectively). This difference in in vivo localization occurred despite the lack of changes in in vitro immunoreactivities of the AM-Fab preparations. The enhanced target localization with minimal hepatic activity may permit a more sensitive diagnostic application of 111 In-labeled AM-Fab in future clinical studies

  12. 75 FR 9438 - Samsung Austin Semiconductor, LLC, DRAM Fab 1, a Subsidiary of Samsung Electronics Corporation...

    Science.gov (United States)

    2010-03-02

    ... Semiconductor, LLC, DRAM Fab 1, a Subsidiary of Samsung Electronics Corporation, Including On-Site Leased... Semiconductor, LLC, a subsidiary of Samsung Electronics Corporation, DRAM Fab 1, including on-site leased.... The workers are engaged in activities related to the production of DRAM chips for use in electronics...

  13. Extending green technology innovations to enable greener fabs

    Science.gov (United States)

    Takahisa, Kenji; Yoo, Young Sun; Fukuda, Hitomi; Minegishi, Yuji; Enami, Tatsuo

    2015-03-01

    Semiconductor manufacturing industry has growing concerns over future environmental impacts as fabs expand and new generations of equipment become more powerful. Especially rare gases supply and price are one of prime concerns for operation of high volume manufacturing (HVM) fabs. Over the past year it has come to our attention that Helium and Neon gas supplies could be unstable and become a threat to HVM fabs. To address these concerns, Gigaphoton has implemented various green technologies under its EcoPhoton program. One of the initiatives is GigaTwin deep ultraviolet (DUV) lithography laser design which enables highly efficient and stable operation. Under this design laser systems run with 50% less electric energy and gas consumption compared to conventional laser designs. In 2014 we have developed two technologies to further reduce electric energy and gas efficiency. The electric energy reduction technology is called eGRYCOS (enhanced Gigaphoton Recycled Chamber Operation System), and it reduces electric energy by 15% without compromising any of laser performances. eGRYCOS system has a sophisticated gas flow design so that we can reduce cross-flow-fan rotation speed. The gas reduction technology is called eTGM (enhanced Total gas Manager) and it improves gas management system optimizing the gas injection and exhaust amount based on laser performances, resulting in 50% gas savings. The next steps in our roadmap technologies are indicated and we call for potential partners to work with us based on OPEN INNOVATION concept to successfully develop faster and better solutions in all possible areas where green innovation may exist.

  14. 20 CFR 30.908 - How will the FAB evaluate new medical evidence submitted to challenge the impairment...

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false How will the FAB evaluate new medical... Medical Evidence of Impairment § 30.908 How will the FAB evaluate new medical evidence submitted to... impairment evaluation that differs from the impairment evaluation relied upon by the district office, the FAB...

  15. Characterization of deamidation at Asn138 in L-chain of recombinant humanized Fab expressed from Pichia pastoris.

    Science.gov (United States)

    Ohkuri, Takatoshi; Murase, Eri; Sun, Shu-Lan; Sugitani, Jun; Ueda, Tadashi

    2013-10-01

    A method was previously established for evaluating Asn deamidation by matrix-assisted laser desorption/ionization time of flight-mass spectrometry using endoproteinase Asp-N. In this study, we demonstrated that this method could be applied to the identification of the deamidation site of the humanized fragment antigen-binding (Fab). First, a system for expressing humanized Fab from methylotrophic yeast Pichia pastoris was constructed, resulting in the preparation of ∼30 mg of the purified humanized Fab from 1 l culture. Analysis of the L-chain derived from recombinant humanized Fab that was heated at pH 7 and 100°C for 1 h showed the deamidation at Asn138 in the constant region. Then, we prepared L-N138D Fab and L-N138A Fab and examined their properties. The circular dichroism (CD) spectrum of the L-N138D Fab was partially different from that of the wild-type Fab. The measurement of the thermostability showed that L-N138D caused a significant decrease in the thermostability of Fab. On the other hand, the CD spectrum and thermostability of L-N138A Fab showed the same behaviour as the wild-type Fab. Thus, it was suggested that the introduction of a negative charge at position 138 in the L-chain by the deamidation significantly affected the stability of humanized Fab.

  16. 20 CFR 30.320 - Can a claim be reopened after the FAB has issued a final decision?

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false Can a claim be reopened after the FAB has... AMENDED Adjudicatory Process Reopening Claims § 30.320 Can a claim be reopened after the FAB has issued a final decision? (a) At any time after the FAB has issued a final decision pursuant to § 30.316, and...

  17. 75 FR 21353 - Intel Corporation, Fab 20 Division, Including On-Site Leased Workers From Volt Technical...

    Science.gov (United States)

    2010-04-23

    ... DEPARTMENT OF LABOR Employment and Training Administration [TA-W-73,642] Intel Corporation, Fab 20... of Intel Corporation, Fab 20 Division, including on-site leased workers of Volt Technical Resources... Precision, Inc. were employed on-site at the Hillsboro, Oregon location of Intel Corporation, Fab 20...

  18. Dynamics of ovarian maturation during the reproductive cycle of Metynnis maculatus, a reservoir invasive fish species (Teleostei: Characiformes

    Directory of Open Access Journals (Sweden)

    Thiago Scremin Boscolo Pereira

    Full Text Available In this study, we evaluated the dynamics of ovarian maturation and the spawning processes during the reproductive cycle of Metynnis maculatus. Adult females (n = 36 were collected bimonthly between April 2010 and March 2011. The mean gonadosomatic index (GSI was determined, ovarian and blood samples were submitted for morphometric evaluation and the steroid plasma concentration was determined by ELISA. This species demonstrated asynchronous ovarian development with multiple spawns. This study revealed that, although defined as a multiple spawning species, the ovaries of M. maculatus have a pattern of development with a predominance of vitellogenesis between April and August and have an intensification in spawning in September; in October, a drop in the mean GSI values occurred, and the highest frequencies of post-ovulatory follicles (POFs were observed. We observed a positive correlation between the POF and the levels of 17α-hydroxyprogesterone. Metynnis maculatus has the potential to be used as a source of pituitary tissue for the preparation of crude extracts for hormonal induction; the theoretical period for use is from September to December, but specific studies to determine the feasibility of this approach must be conducted.

  19. Effect of Gamma Rays on Some Aspects of Biological Performance of Callosobrchus maculatus (Fabricius) and Trogoderma granarium (Everts)

    International Nuclear Information System (INIS)

    Al-Taweel, A.A.; Al-Jowary, S.A.; Mahmmod, E.A.

    2007-01-01

    The effect of gamma rays on males and females of Callasobruchus maculatus and Trogoderma granarium which were irradiated as 1-3 days old adults was investigated.The results revealed that the percent egg hatch for both pests was zero, and average number of egg (34.2, 21.5) for both pests respectively where their males where irradiated with 0.18 KGy and mated with unirradiated females.While the percent of egg hatch and the average number of egg (zero, 21, 3) respectively when the females where irradiated with 0.18 and 0.15 KGy and mated to unirradiated males for C. maculatus and T. granarium respectively.Furthermore, the results showed that the percent of eggs hatch and average number of egg was (zero,22.7) for C. maculatus when both sexes irradiated with 0.12 KGy and mated together and it was also (zero, 11.5) for T. granarium when both sexes where irradiated with 0.15 KGy mated together

  20. Locking the Elbow: Improved Antibody Fab Fragments as Chaperones for Structure Determination.

    Science.gov (United States)

    Bailey, Lucas J; Sheehy, Kimberly M; Dominik, Pawel K; Liang, Wenguang G; Rui, Huan; Clark, Michael; Jaskolowski, Mateusz; Kim, Yejoon; Deneka, Dawid; Tang, Wei-Jen; Kossiakoff, Anthony A

    2018-02-02

    Antibody Fab fragments have been exploited with significant success to facilitate the structure determination of challenging macromolecules as crystallization chaperones and as molecular fiducial marks for single particle cryo-electron microscopy approaches. However, the inherent flexibility of the "elbow" regions, which link the constant and variable domains of the Fab, can introduce disorder and thus diminish their effectiveness. We have developed a phage display engineering strategy to generate synthetic Fab variants that significantly reduces elbow flexibility, while maintaining their high affinity and stability. This strategy was validated using previously recalcitrant Fab-antigen complexes where introduction of an engineered elbow region enhanced crystallization and diffraction resolution. Furthermore, incorporation of the mutations appears to be generally portable to other synthetic antibodies and may serve as a universal strategy to enhance the success rates of Fabs as structure determination chaperones. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. The Emerging Importance of IgG Fab Glycosylation in Immunity.

    Science.gov (United States)

    van de Bovenkamp, Fleur S; Hafkenscheid, Lise; Rispens, Theo; Rombouts, Yoann

    2016-02-15

    Human IgG is the most abundant glycoprotein in serum and is crucial for protective immunity. In addition to conserved IgG Fc glycans, ∼15-25% of serum IgG contains glycans within the variable domains. These so-called "Fab glycans" are primarily highly processed complex-type biantennary N-glycans linked to N-glycosylation sites that emerge during somatic hypermutation. Specific patterns of Fab glycosylation are concurrent with physiological and pathological conditions, such as pregnancy and rheumatoid arthritis. With respect to function, Fab glycosylation can significantly affect stability, half-life, and binding characteristics of Abs and BCRs. Moreover, Fab glycans are associated with the anti-inflammatory activity of IVIgs. Consequently, IgG Fab glycosylation appears to be an important, yet poorly understood, process that modulates immunity. Copyright © 2016 by The American Association of Immunologists, Inc.

  2. Isolation of Osteosarcoma-Associated Human Antibodies from a Combinatorial Fab Phage Display Library

    Directory of Open Access Journals (Sweden)

    Carmela Dantas-Barbosa

    2009-01-01

    Full Text Available Osteosarcoma, a highly malignant disease, is the most common primary bone tumor and is frequently found in children and adolescents. In order to isolate antibodies against osteosarcoma antigens, a combinatorial osteosarcoma Fab library displayed on the surface of phages was used. After three rounds of selection on the surface of tumor cells, several osteosarcoma-reactive Fabs were detected. From these Fabs, five were better characterized, and despite having differences in their VH (heavy chain variable domain and Vκ (kappa chain variable domain regions, they all bound to a protein with the same molecular mass. Further analysis by cell ELISA and immunocytochemistry suggested that the Fabs recognize a membrane-associated tumor antigen expressed in higher amounts in neoplasic cells than in normal tissue. These results suggest that the human Fabs selected in this work are a valuable tool for the study of this neoplasia.

  3. Optimal expression of a Fab-effector fusion protein in Escherichia coli by removing the cysteine residues responsible for an interchain disulfide bond of a Fab molecule.

    Science.gov (United States)

    Kang, Hyeon-Ju; Kim, Hye-Jin; Jung, Mun-Sik; Han, Jae-Kyu; Cha, Sang-Hoon

    2017-04-01

    Development of novel bi-functional or even tri-functional Fab-effector fusion proteins would have a great potential in the biomedical sciences. However, the expression of Fab-effector fusion proteins in Escherichia coli is problematic especially when a eukaryotic effector moiety is genetically linked to a Fab due to the lack of proper chaperone proteins and an inappropriate physicochemical environment intrinsic to the microbial hosts. We previously reported that a human Fab molecule, referred to as SL335, reactive to human serum albumin has a prolonged in vivo serum half-life in rats. We, herein, tested six discrete SL335-human growth hormone (hGH) fusion constructs as a model system to define an optimal Fab-effector fusion format for E. coli expression. We found that one variant, referred to as HserG/Lser, outperformed the others in terms of a soluble expression yield and functionality in that HserG/Lser has a functional hGH bioactivity and possesses an serum albumin-binding affinity comparable to SL335. Our results clearly demonstrated that the genetic linkage of an effector domain to the C-terminus of Fd (V H +C H1 ) and the removal of cysteine (Cys) residues responsible for an interchain disulfide bond (IDB) ina Fab molecule optimize the periplasmic expression of a Fab-effector fusion protein in E. coli. We believe that our approach can contribute the development of diverse bi-functional Fab-effector fusion proteins by providing a simple strategy that enables the reliable expression of a functional fusion proteins in E. coli. Copyright © 2017 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.

  4. Fab-based bispecific antibody formats with robust biophysical properties and biological activity.

    Science.gov (United States)

    Wu, Xiufeng; Sereno, Arlene J; Huang, Flora; Lewis, Steven M; Lieu, Ricky L; Weldon, Caroline; Torres, Carina; Fine, Cody; Batt, Micheal A; Fitchett, Jonathan R; Glasebrook, Andrew L; Kuhlman, Brian; Demarest, Stephen J

    2015-01-01

    A myriad of innovative bispecific antibody (BsAb) platforms have been reported. Most require significant protein engineering to be viable from a development and manufacturing perspective. Single-chain variable fragments (scFvs) and diabodies that consist only of antibody variable domains have been used as building blocks for making BsAbs for decades. The drawback with Fv-only moieties is that they lack the native-like interactions with CH1/CL domains that make antibody Fab regions stable and soluble. Here, we utilize a redesigned Fab interface to explore 2 novel Fab-based BsAbs platforms. The redesigned Fab interface designs limit heavy and light chain mixing when 2 Fabs are co-expressed simultaneously, thus allowing the use of 2 different Fabs within a BsAb construct without the requirement of one or more scFvs. We describe the stability and activity of a HER2×HER2 IgG-Fab BsAb, and compare its biophysical and activity properties with those of an IgG-scFv that utilizes the variable domains of the same parental antibodies. We also generated an EGFR × CD3 tandem Fab protein with a similar format to a tandem scFv (otherwise known as a bispecific T cell engager or BiTE). We show that the Fab-based BsAbs have superior biophysical properties compared to the scFv-based BsAbs. Additionally, the Fab-based BsAbs do not simply recapitulate the activity of their scFv counterparts, but are shown to possess unique biological activity.

  5. Radioimmunoimaging using F(ab')2 fragment of monoclonal antibodies against human alpha-fetoprotein

    International Nuclear Information System (INIS)

    Sakahara, Harumi; Endo, Keigo; Nakashima, Tetsuo; Koizumi, Mitsuru; Ohta, Hitoya; Torizuka, Kanji; Okada, Kenichiro; Yoshida, Osamu; Nishi, Shinzo.

    1985-01-01

    Using monoclonal antibodies against human α-fetoprotein (AFP), radioiodinated F(ab') 2 fragments were compared with whole IgG as a radiotracer for radioimmunoimaging of cancer. F(ab') 2 fragments were obtained by pepsin digestion of whole IgG (IgGl). IgG and F(ab') 2 were labeled with 125 I or 131 I by the chloramine-T method with almost full retention of antibody activity. F(ab') 2 fragments were cleared more rapidly from the circulation in normal mice with a half life of 6.3 hours than whole IgG with a half life of 5.5 days. Radioactivity of F(ab') 2 in various organs also decreased faster than IgG. In nude mice transplanted with AFP-producing human testicular tumor, F(ab') 2 fragments demonstrated superior scintigrams to whole IgG at 2 days after the injection, because of the fast disappearance of background radioactivity. Although absolute accumulation of 131 I labeled F(ab') 2 in the tumor was less than that of 131 I labeled IgG, tumor to other organ ratios were much higher with F(ab') 2 than those of IgG. The tumor to blood ratio of 131 I labeled F(ab') 2 was 1.04 at day 2, whereas tumor to blood ratio of 131 I labeled IgG was 0.55 at day 2 and 0.92 at day 4, respectively. These results indicated that for the radiolabeling of monoclonal antibodies, F(ab') 2 fragments would be superior to whole IgG in the radioimmunoimaging of cancer. (author)

  6. Clinical usefulness of human-mouse chimeric Fab monoclonal antibody A7 for radioimmunoguided surgery

    International Nuclear Information System (INIS)

    Yamamoto, Kazuhito

    1999-01-01

    This study was designed to determine the clinical usefulness of radioimmunoguided surgery (RIGS) using the human-mouse chimeric Fab monoclonal antibody A7 (chA7Fab) for colorectal cancer patients. Whole murine monoclonal antibody A7 (whole A7) and chA7Fab were labelled with 125 I and 131 I, and their biodistributions were investigated experimentally and clinically. Radioactivities of the antibodies in the tissues were measured by a portable gamma detecting probe (GDP) purchased from Neoprobe Corp.. Of the four labelled antibodies used in a mouse model, 125 I-chA7Fab revealed the highest tumor/surrounding tissue ratio and all values were greater than 2.0. All tumor/surrounding tissue ratios of 131 I-chA7Fab were greater than 1.5, but the values were lower than those of 125 I-chA7Fab. Due to the limited clinical use of 125 I in Japan, 131 I was used as a radio-tracer for chA7Fab in the clinical trial. RIGS using 131 I-chA7Fab was performed on ten colorectal cancer patients. Tumor localization was intraoperatively determined in four of ten patients using the GDP. Liver metastasis and lymph node metastasis were identified in two patients and one patient, respectively. The GDP revealed tumor/surrounding tissue ratios of 1.5 or greater in eight of the ten resected tumors. Although radioimmunoguided surgery using chA7Fab is a promising tool to intraoperatively determine the tumor localization of colorectal cancer, 125 I and not 131 I should be used as a tracer for radioimmunoguided surgery to increase the accuracy of chA7Fab. (author)

  7. Comparing domain interactions within antibody Fabs with kappa and lambda light chains.

    Science.gov (United States)

    Toughiri, Raheleh; Wu, Xiufeng; Ruiz, Diana; Huang, Flora; Crissman, John W; Dickey, Mark; Froning, Karen; Conner, Elaine M; Cujec, Thomas P; Demarest, Stephen J

    2016-10-01

    IgG antibodies are multi-domain proteins with complex inter-domain interactions. Human IgG heavy chains (HCs) associate with light chains (LCs) of the κ or λ isotype to form mature antibodies capable of binding antigen. The HC/LC interaction involves 4 domains: VH and CH1 from the HC and VL and CL from the LC. Human Fabs with κ LCs have been well characterized for their unfolding behaviors and demonstrate a significant level of cooperativity and stabilization when all 4 domains are intact. Very little is known regarding the thermodynamic properties of human Fabs with λ LCs. Here, we dissect the domain contributions to Fab stability for both κ and λ LC-containing Fabs. We find the cooperativity of unfolding between the constant domains, CH1/Cλ, and variable domains, VH/Vλ, within λ LC-containing Fabs is significantly weaker than that of κ LC-containing Fabs. The data suggests there may not be an evolutionary necessity for strong variable/constant domain cooperativity within λ LC-containing Fabs. After investigating the biophysical properties of Fabs with mismatched variable and constant domain subunits (e.g., VH/Vκ paired with CH1/Cλ or T cell receptor Cα/Cβ), the major role of the constant domains for both κ- and λ-containing Fabs may be to reduce the hydrophobic exposure at the VH/VL interface. Even though Fabs with these non-native pairings were thermodynamically less stable, they secreted well from mammalian cells as well behaved monodisperse proteins, which was in contrast to what was observed with the VH/Vκ and VH/Vλ scFvs that secreted as a mixture of monomer and aggregates.

  8. Subzero water permeability parameters and optimal freezing rates for sperm cells of the southern platyfish, Xiphophorus maculatus.

    Science.gov (United States)

    Pinisetty, D; Huang, C; Dong, Q; Tiersch, T R; Devireddy, R V

    2005-06-01

    This study reports the subzero water transport characteristics (and empirically determined optimal rates for freezing) of sperm cells of live-bearing fishes of the genus Xiphophorus, specifically those of the southern platyfish Xiphophorus maculatus. These fishes are valuable models for biomedical research and are commercially raised as ornamental fish for use in aquariums. Water transport during freezing of X. maculatus sperm cell suspensions was obtained using a shape-independent differential scanning calorimeter technique in the presence of extracellular ice at a cooling rate of 20 degrees C/min in three different media: (1) Hanks' balanced salt solution (HBSS) without cryoprotective agents (CPAs); (2) HBSS with 14% (v/v) glycerol, and (3) HBSS with 10% (v/v) dimethyl sulfoxide (DMSO). The sperm cell was modeled as a cylinder with a length of 52.35 microm and a diameter of 0.66 microm with an osmotically inactive cell volume (Vb) of 0.6 V0, where V0 is the isotonic or initial cell volume. This translates to a surface area, SA to initial water volume, WV ratio of 15.15 microm(-1). By fitting a model of water transport to the experimentally determined volumetric shrinkage data, the best fit membrane permeability parameters (reference membrane permeability to water at 0 degrees C, Lpg or Lpg [cpa] and the activation energy, E(Lp) or E(Lp) [cpa]) were found to range from: Lpg or Lpg [cpa] = 0.0053-0.0093 microm/minatm; E(Lp) or E(Lp) [cpa] = 9.79-29.00 kcal/mol. By incorporating these membrane permeability parameters in a recently developed generic optimal cooling rate equation (optimal cooling rate, [Formula: see text] where the units of B(opt) are degrees C/min, E(Lp) or E(Lp) [cpa] are kcal/mol, L(pg) or L(pg) [cpa] are microm/minatm and SA/WV are microm(-1)), we determined the optimal rates of freezing X. maculatus sperm cells to be 28 degrees C/min (in HBSS), 47 degrees C/min (in HBSS+14% glycerol) and 36 degrees C/min (in HBSS+10% DMSO). Preliminary empirical

  9. Defect reduction in different periods of a fab's life

    Science.gov (United States)

    Kallis, Norbert

    1997-08-01

    The reduction of yield loss caused by particles is a key topic of productivity enhancement. It deals with many components of a fab's structure: (1) Cultural mindset of managers, engineers, and operators -- to have champions for that mission, (2) selection of defect metrology tools -- with high throughput and reliable data, (3) single equipment characterization -- to set challenging specs according to the best performing tool, (4) inline reaction procedures -- to transfer knowledge and responsibility to the operators, (5) statistical evaluation of large datasets -- including correlation to electrical yield, (6) and to trace back the way to the single tools or the low performing processes. This paper discusses some topics to be considered during process and equipment planning, ramp up, and mature production. It has been proven to be successful during a very aggressive ramp up in wafer starts and an even more convincing yield increase of a fab, that now started the production of DRAM's in quarter micron technology. It will conclude in an understanding, that besides a reliable statistical methodology the human performance is the key: can we really integrate people of different groups and levels of responsibility and motivate them to take joint actions for yield goals?

  10. Radioactive tracers in the study of energy turnover by a grazing insect (Chrysochus auratus Fab. ; coleoptera chrysomelidae)

    Energy Technology Data Exchange (ETDEWEB)

    Williams, Jr, E C; Reichle, D E

    1968-01-01

    Radioisotope tags of rubidium-86 and phosphorus-32 were used to measure food consumption by the chrysomelid beetle Chrysochus auratus feeding upon the old field herb Apocynum cannabinum. The elimination of isotope by beetles which had fed upon tagged plants was used to estimate the rates of food consumption. For individuals at isotopic equilibrium with their food base, isotope input (I) equals the biological turnover of isotope: I = kQ/sub e/M/a, where k is the turnover rate of isotope, Q/sub e/ is the equilibrium concentration of isotope in the beetles, M is mass (mg dry wt), and a is the assimilation coefficient for the particular radioisotope. Turnover rates (k) were calculated from biological half-lives (T/sub b/), where k = 0.693/T/sub b/. The biological half-life of YWRb in Chrysochus was one day, while that of TSP was 7.5 days in females and 10.2 days in males. The calorific value of Apocynum leaves was 4625 cal/g dry wt (5640 cal/g ash-free wt); that of Chrysochus was 5227 cal/g dry wt (5537 cal/g ash-free wt). Individuals consumed 56.8% (15.9 mg) of their dry body weight in food per day. Of the ingested foliage, 43% of the ash-free dry matter was digested, 56% of the energy assimilated, and 71% of the ash content assimilated. The calorific value (cal/g) of digested food was 1.17 times greater than that consumed. Beetles assimilated 87% of the YWRb and 74% of the TSP present in food. Ranking of turnover times showed Chrysochus to be most conservative in its expenditure of food energy and dry matter, followed by mineral constituents. 9 references, 3 figures, 3 tables.

  11. Structural and dynamical aspects of Streptococcus gordonii FabH through molecular docking and MD simulations.

    Science.gov (United States)

    Shamim, Amen; Abbasi, Sumra Wajid; Azam, Syed Sikander

    2015-07-01

    β-Ketoacyl-ACP-synthase III (FabH or KAS III) has become an attractive target for the development of new antibacterial agents which can overcome the multidrug resistance. Unraveling the fatty acid biosynthesis (FAB) metabolic pathway and understanding structural coordinates of FabH will provide valuable insights to target Streptococcus gordonii for curing oral infection. In this study, we designed inhibitors against therapeutic target FabH, in order to block the FAB pathway. As compared to other targets, FabH has more interactions with other proteins, located on the leading strand with higher codon adaptation index value and associated with lipid metabolism category of COG. Current study aims to gain in silico insights into the structural and dynamical aspect of S. gordonii FabH via molecular docking and molecular dynamics (MD) simulations. The FabH protein is catalytically active in dimerization while it can lock in monomeric state. Current study highlights two residues Pro88 and Leu315 that are close to each other by dimerization. The active site of FabH is composed of the catalytic triad formed by residues Cys112, His249, and Asn279 in which Cys112 is involved in acetyl transfer, while His249 and Asn279 play an active role in decarboxylation. Docking analysis revealed that among the studied compounds, methyl-CoA disulfide has highest GOLD score (82.75), binding affinity (-11 kcal/mol) and exhibited consistently better interactions. During MD simulations, the FabH structure remained stable with the average RMSD value of 1.7 Å and 1.6 Å for undocked protein and docked complex, respectively. Further, crucial hydrogen bonding of the conserved catalytic triad for exhibiting high affinity between the FabH protein and ligand is observed by RDF analysis. The MD simulation results clearly demonstrated that binding of the inhibitor with S. gordonii FabH enhanced the structure and stabilized the dimeric FabH protein. Therefore, the inhibitor has the potential to become

  12. The response of Dendroctonus valens (Coleoptera: Scolytidae) and Temnochila chlorodia (Coleoptera: Trogossitidae) to Ips paraconfusus (Coleoptera: Scolytidae) pheromone components and verbenone

    Science.gov (United States)

    Christopher J. Fettig; Stepehen R. McKelvey; Christopher P. Dabney; Robert R. Borys

    2007-01-01

    The red turpentine beetle, Dendroctonus valens LeConte, 1860 (Coleoptera: Curculionidae, Scolytinae), is a common bark beetle species found throughout much of North America and China. In 2004, we observed that California fivespined ips, Ips paraconfusus Lanier, 1970 (Coleoptera: Curculionidae, Scolytinae), attack densities in logging debris were inversely related to D...

  13. FabQ, a Dual-Function Dehydratase/Isomerase, Circumvents the Last Step of the Classical Fatty Acid Synthesis Cycle

    OpenAIRE

    Bi, Hongkai; Wang, Haihong; Cronan, John E.

    2013-01-01

    In the classical anaerobic pathway of unsaturated fatty acid biosynthesis, that of Escherichia coli, the double bond is introduced into the growing acyl chain by the FabA dehydratase/isomerase. Another dehydratase, FabZ, functions in the chain elongation cycle. In contrast, Aerococcus viridans has only a single FabA/FabZ homolog we designate FabQ. FabQ can not only replace the function of E. coli FabZ in vivo, but it also catalyzes the isomerization required for unsaturated fatty acid biosynt...

  14. Meleagrin, a new FabI inhibitor from Penicillium chryosogenum with at least one additional mode of action.

    Directory of Open Access Journals (Sweden)

    Chang Ji Zheng

    Full Text Available Bacterial enoyl-acyl carrier protein reductase (FabI is a promising novel antibacterial target. We isolated a new class of FabI inhibitor from Penicillium chrysogenum, which produces various antibiotics, the mechanisms of some of them are unknown. The isolated FabI inhibitor was determined to be meleagrin by mass spectroscopy and nuclear magnetic resonance spectral analyses, and its more active and inactive derivatives were chemically prepared. Consistent with their selective inhibition of Staphylococcus aureus FabI, meleagrin and its more active derivatives directly bound to S. aureus FabI in a fluorescence quenching assay, inhibited intracellular fatty acid biosynthesis and growth of S. aureus, and increased the minimum inhibitory concentration for fabI-overexpressing S. aureus. The compounds that were not effective against the FabK isoform, however, inhibited the growth of Streptococcus pneumoniae that contained only the FabK isoform. Additionally no resistant mutant to the compounds was obtained. Importantly, fabK-overexpressing Escherichia coli was not resistant to these compounds, but was resistant to triclosan. These results demonstrate that the compounds inhibited another target in addition to FabI. Thus, meleagrin is a new class of FabI inhibitor with at least one additional mode of action that could have potential for treating multidrug-resistant bacteria.

  15. Immobilization of Fab' fragments onto substrate surfaces: A survey of methods and applications.

    Science.gov (United States)

    Crivianu-Gaita, Victor; Thompson, Michael

    2015-08-15

    Antibody immobilization onto surfaces has widespread applications in many different fields. It is desirable to bind antibodies such that their fragment-antigen-binding (Fab) units are oriented away from the surface in order to maximize analyte binding. The immobilization of only Fab' fragments yields benefits over the more traditional whole antibody immobilization technique. Bound Fab' fragments display higher surface densities, yielding a higher binding capacity for the analyte. The nucleophilic sulfide of the Fab' fragments allows for specific orientations to be achieved. For biosensors, this indicates a higher sensitivity and lower detection limit for a target analyte. The last thirty years have shown tremendous progress in the immobilization of Fab' fragments onto gold, Si-based, polysaccharide-based, plastic-based, magnetic, and inorganic surfaces. This review will show the current scope of Fab' immobilization techniques available and illustrate methods employed to minimize non-specific adsorption of undesirables. Furthermore, a variety of examples will be given to show the versatility of immobilized Fab' fragments in different applications and future directions of the field will be addressed, especially regarding biosensors. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Fab glycosylation of immunoglobulin G does not associate with improvement of rheumatoid arthritis during pregnancy.

    Science.gov (United States)

    Bondt, Albert; Wuhrer, Manfred; Kuijper, T Martijn; Hazes, Johanna M W; Dolhain, Radboud J E M

    2016-11-25

    Changes in immunoglobulin G (IgG) constant domain (Fc) glycosylation are associated with changes in rheumatoid arthritis (RA) disease activity in response to pregnancy. Here, we sought to determine whether the same holds true for variable domain (Fab) glycosylation. IgGs were captured from RA and control sera obtained before (RA only), during and after pregnancy, followed by Fc and Fab separation, glycan release, and mass spectrometric detection. In parallel, glycans from intact IgG were analysed. The data was used to calculate glycosylation traits, and to estimate the level of Fab glycosylation. The overall level of Fab glycosylation was increased in RA patients compared to controls, while no differences in Fab glycosylation patterns were found. For the Fc and intact IgG (Total) previously observed differences in galactosylation and bisection were confirmed. Furthermore, increased galactosylation of Fc and Total were associated with lower disease activity and autoantibody positivity. In addition, the change in Fc galactosylation associated with the change in disease activity during pregnancy and after delivery, while this was not the case for Fab. In contrast to changes in Fc glycosylation, changes in Fab glycosylation are not associated with improvement of RA during pregnancy and arthritis flare after delivery.

  17. 'Zipbody' leucine zipper-fused Fab in E. coli in vitro and in vivo expression systems.

    Science.gov (United States)

    Ojima-Kato, Teruyo; Fukui, Kansuke; Yamamoto, Hiroaki; Hashimura, Dai; Miyake, Shiro; Hirakawa, Yuki; Yamasaki, Tomomi; Kojima, Takaaki; Nakano, Hideo

    2016-04-01

    A small antibody fragment, fragment of antigen binding (Fab), is favorable for various immunological assays. However, production efficiency of active Fab in microorganisms depends considerably on the clones. In this study, leucine zipper-peptide pairs that dimerize in parallel (ACID-p1 (LZA)/BASE-p1 (LZB) or c-Jun/c-Fos) were fused to the C-terminus of heavy chain (Hc, VH-CH1) and light chain (Lc, VL-CL), respectively, to accelerate the association of Hc and Lc to form Fab in Escherichia coli in vivo and in vitro expression systems. The leucine zipper-fused Fab named 'Zipbody' was constructed using anti-E. coli O157 monoclonal antibody obtained from mouse hybridoma and produced in both in vitro and in vivo expression systems in an active form, whereas Fab without the leucine zipper fusion was not. Similarly, Zipbody of rabbit monoclonal antibody produced in in vitro expression showed significant activity. The purified, mouse Zipbody produced in the E. coli strain Shuffle T7 Express had specificity toward the antigen; in bio-layer interferometry analysis, the KD value was measured to be 1.5-2.0 × 10(-8) M. These results indicate that leucine zipper fusion to Fab C-termini markedly enhances active Fab formation in E. coli. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  18. Isolation of human anti-serum albumin Fab antibodies with an extended serum-half life.

    Science.gov (United States)

    Kang, Hyeon-Ju; Kim, Hye-Jin; Cha, Sang-Hoon

    2016-01-01

    The serum albumin (SA) has been exploited to generate long-acting biotherapeutics by taking advantage of the FcRn-mediated recycling mechanism in a direct or an indirect way. Since Fab fragments have been proven to be clinically safe for human usage, we assumed that human anti-SA Fab antibodies could have a great potential as a carrier molecule to extend the serum half-life of therapeutic proteins. We, herein, had attempted to isolate anti-SA Fab antibodies from HuDVFab-8L antibody library via a phage display technology, and identified eight discrete human Fab antibodies. One of the Fab antibodies, SL335, showed the strongest binding reactivity to human SA with nM range of affinity at both pH 6 and pH 7.4, and cross-reacted to SAs from various species including rat, mouse, canine and monkey. The in vivo pharmacokinetic assay using a rat model indicated that SL335 has approximately 10 fold longer serum half-life and 26 to 44-fold increase in AUC0 → ∞ compared to the negative control Fab molecule in both intravenous and subcutaneous administrations. Knowing that Fabs have proven to be safe in clinics for a long time, SL335 seems to have a great potential in generating long-acting protein drugs by tagging effector molecules with either chemical conjugation or genetic fusion. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Structural features of Fab fragments of rheumatoid factor IgM-RF in solution

    International Nuclear Information System (INIS)

    Volkov, V. V.; Lapuk, V. A.; Shtykova, E. V.; Stepina, N. D.; Dembo, K. A.; Sokolova, A. V.; Amarantov, S. V.; Timofeev, V. P.; Ziganshin, R. Kh.; Varlamova, E. Yu.

    2008-01-01

    The structural features of the Fab fragments of monoclonal (Waldenstroem's disease) immunoglobulin M (IgM) and rheumatoid immunoglobulin M (IgM-RF) were studied by a complex of methods, including small-angle X-ray scattering (SAXS), electron spin resonance (ESR), and mass spectrometry (MS). The Fab-RF fragment was demonstrated to be much more flexible in the region of interdomain contacts, the molecular weights and the shapes of the Fab and Fab-RF macromolecules in solution being only slightly different. According to the ESR data, the rotational correlation time for a spin label introduced into the peptide sequence for Fab is twice as large as that for Fab-RF (21±2 and 11±1 ns, respectively), whereas the molecular weights of these fragments differ by only 0.5% (mass-spectrometric data), which correlates with the results of molecular-shape modeling by small-angle X-ray scattering. The conclusion about the higher flexibility of the Fab-RF fragment contributes to an understanding of the specificity of interactions between the rheumatoid factor and the antigens of the own organism.

  20. Hepatic targeting and hypocholesterolemic effect of lactosaminated Fab against low density lipoproteins

    International Nuclear Information System (INIS)

    Bernini, F.; Bocan, T.M.A.; Via, D.P.; Gotto, A.M. Jr.; Smith, L.C.

    1986-01-01

    Lactosaminated Fab (lac-Fab) specific for human LDL induces plasma clearance and uptake of circulating [ 125 -I]-iodo-LDL in rat, a process mediated by galactose receptors of the liver. This study demonstrates that lac-Fab is a specific carrier of LDL to the liver parenchymal cells and exhibits hypocholesterolemic activity in vivo. Rats were injected with fluorescent 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine-LDL (diI-LDL) or 6 mg of LDL plus tracer amounts of [ 125 I]-iodo-LDL. After 10-20 min, the animals received 3-10 mg of lac-Fab. Histologic examination of the liver sections showed the uptake of diI-LDL in the parenchymal cells, as compared to diI-acetyl-LDL which was localized in sinusoidal cells. More than 85% of human LDL disappeared within 2.5 hr after lac-Fab injection, reducing plasma cholesterol from 133.0 +/- 12.6 mg/dl to 66.4 +/- 8.0 mg/dl, the basal value in the rat. In control rats, only about 20% of radioactivity and cholesterol disappeared at 2.5 hr. HDL levels were unaffected. The authors conclude that lac-Fab is a specific carrier of LDL to hepatocytes and can lower plasma LDL-cholesterol in vivo. Lac-Fab specific for other antigens may act as specific carriers of molecule or macromolecules to hepatocytes

  1. The community FabLab platform: applications and implications in biomedical engineering.

    Science.gov (United States)

    Stephenson, Makeda K; Dow, Douglas E

    2014-01-01

    Skill development in science, technology, engineering and math (STEM) education present one of the most formidable challenges of modern society. The Community FabLab platform presents a viable solution. Each FabLab contains a suite of modern computer numerical control (CNC) equipment, electronics and computing hardware and design, programming, computer aided design (CAD) and computer aided machining (CAM) software. FabLabs are community and educational resources and open to the public. Development of STEM based workforce skills such as digital fabrication and advanced manufacturing can be enhanced using this platform. Particularly notable is the potential of the FabLab platform in STEM education. The active learning environment engages and supports a diversity of learners, while the iterative learning that is supported by the FabLab rapid prototyping platform facilitates depth of understanding, creativity, innovation and mastery. The product and project based learning that occurs in FabLabs develops in the student a personal sense of accomplishment, self-awareness, command of the material and technology. This helps build the interest and confidence necessary to excel in STEM and throughout life. Finally the introduction and use of relevant technologies at every stage of the education process ensures technical familiarity and a broad knowledge base needed for work in STEM based fields. Biomedical engineering education strives to cultivate broad technical adeptness, creativity, interdisciplinary thought, and an ability to form deep conceptual understanding of complex systems. The FabLab platform is well designed to enhance biomedical engineering education.

  2. Single-reagent one-step procedures for the purification of ovine IgG, F(ab')2 and Fab antivenoms by caprylic acid.

    Science.gov (United States)

    Al-Abdulla, Ibrahim; Casewell, Nicholas R; Landon, John

    2014-01-15

    Antivenoms are typically produced in horses or sheep and often purified using salt precipitation of immunoglobulins or F(ab')2 fragments. Caprylic (octanoic) acid fractionation of antiserum has the advantage of not precipitating the desired antibodies, thereby avoiding potential degradation that can lead to the formation of aggregates, which may be the cause of some adverse reactions to antivenoms. Here we report that when optimising the purification of immunoglobulins from ovine antiserum raised against snake venom, caprylic acid was found to have no effect on the activity of the enzymes pepsin and papain, which are employed in antivenom manufacturing to digest immunoglobulins to obtain F(ab')2 and Fab fragments, respectively. A "single-reagent" method was developed for the production of F(ab')2 antivenom whereby whole ovine antiserum was mixed with both caprylic acid and pepsin and incubated for 4h at 37°C. For ovine Fab antivenom production from whole antiserum, the "single reagent" comprised of caprylic acid, papain and l-cysteine; after incubation at 37°C for 18-20h, iodoacetamide was added to stop the reaction. Caprylic acid facilitated the precipitation of albumin, resulting in a reduced protein load presented to the digestion enzymes, culminating in substantial reductions in processing time. The ovine IgG, F(ab')2 and Fab products obtained using these novel caprylic acid methods were comparable in terms of yield, purity and specific activity to those obtained by multi-step conventional salt fractionation with sodium sulphate. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Fab fragments of ovine antibody to colchicine enhance its clearance in the rat.

    Science.gov (United States)

    Peake, Philip W; Pianta, Timothy J; Succar, Lena; Fernando, Mangalee; Buckley, Nicholas A; Endre, Zoltan H

    2015-06-01

    Colchicine is an anti-inflammatory alkaloid used for the treatment of acute gout, but has a narrow therapeutic index. Colchicine overdoses are relatively rare, but have high mortality requiring rapid treatment. To evaluate the ability of a newly available ovine fragment antigen-binding (Fab) antibody to colchicine (ColchiFab(™)) to protect rats against renal and other injury 24 h after colchicine ingestion. Rats were gavaged with colchicine (5 mg/kg), then 2 h later injected intraperitoneally with 5 ml of sterile saline, or Fab anti-colchicine, a newly available ovine antibody to colchicine. Samples of blood were taken at 1, 2, 5 and 24 h after gavage, and urine was collected from 5 to 24 h after gavage. Concentrations of colchicine in tissue, blood and urine were measured by liquid chromatography/mass spectrometry, concentrations of Fab anti-colchicine, urinary neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 or KIM-1 by enzyme-linked immunosorbent assay or ELISA, while concentrations of creatine kinase and creatinine (Cr) were measured enzymatically. Colchicine equilibrated rapidly throughout the body and increased serum creatine kinase. Fab anti-colchicine also rapidly redistributed to the blood and remained at high concentrations over 24 h. Fab anti-colchicine caused a rapid 7.1-fold increase in serum colchicine level, followed by excretion of both colchicine and Fab anti-colchicine through the urine. This was associated with the accumulation of colchicine in the kidney, a reversal of colchicine-induced diarrhoea, and increasing urinary NGAL level; from 168 ± 48 to 477 ± 255 ng/mmol Cr [mean ± standard deviation or SD]. Fab anti-colchicine greatly increased the clearance of colchicine, although increasing NGAL level suggested the presence of mild kidney damage. These data suggest clinical utility for Fab anti-colchicine in the treatment of colchicine overdose.

  4. Site-specific fab fragment biotinylation at the conserved nucleotide binding site for enhanced Ebola detection.

    Science.gov (United States)

    Mustafaoglu, Nur; Alves, Nathan J; Bilgicer, Basar

    2015-07-01

    The nucleotide binding site (NBS) is a highly conserved region between the variable light and heavy chains at the Fab domains of all antibodies, and a small molecule that we identified, indole-3-butyric acid (IBA), binds specifically to this site. Fab fragment, with its small size and simple production methods compared to intact antibody, is good candidate for use in miniaturized diagnostic devices and targeted therapeutic applications. However, commonly used modification techniques are not well suited for Fab fragments as they are often more delicate than intact antibodies. Fab fragments are of particular interest for sensor surface functionalization but immobilization results in damage to the antigen binding site and greatly reduced activity due to their truncated size that allows only a small area that can bind to surfaces without impeding antigen binding. In this study, we describe an NBS-UV photocrosslinking functionalization method (UV-NBS(Biotin) in which a Fab fragment is site-specifically biotinylated with an IBA-EG11-Biotin linker via UV energy exposure (1 J/cm(2)) without affecting its antigen binding activity. This study demonstrates successful immobilization of biotinylated Ebola detecting Fab fragment (KZ52 Fab fragment) via the UV-NBS(Biotin) method yielding 1031-fold and 2-fold better antigen detection sensitivity compared to commonly used immobilization methods: direct physical adsorption and NHS-Biotin functionalization, respectively. Utilization of the UV-NBS(Biotin) method for site-specific conjugation to Fab fragment represents a proof of concept use of Fab fragment for various diagnostic and therapeutic applications with numerous fluorescent probes, affinity molecules and peptides. © 2015 Wiley Periodicals, Inc.

  5. Family of Advanced Beyond Line-of-Sight Terminals (FAB-T)

    Science.gov (United States)

    2015-12-01

    Selected Acquisition Report (SAR) RCS: DD-A&T(Q&A)823-199 Family of Advanced Beyond Line-of-Sight Terminals ( FAB -T) As of FY 2017 President’s...Budget Defense Acquisition Management Information Retrieval (DAMIR) March 21, 2016 15:24:15 UNCLASSIFIED FAB -T December 2015 SAR March 21, 2016 15:24...Operational Requirements Document OSD - Office of the Secretary of Defense O&S - Operating and Support PAUC - Program Acquisition Unit Cost FAB -T December 2015

  6. Cationization increases brain distribution of an amyloid-beta protofibril selective F(ab')2 fragment

    OpenAIRE

    Syvänen, Stina; Edén, Desireé; Sehlin, Dag

    2017-01-01

    Antibodies and fragments thereof are, because of high selectivity for their targets, considered as potential therapeutics and biomarkers for several neurological disorders. However, due to their large molecular size, antibodies/fragments do not easily penetrate into the brain. The aim of the present study was to improve the brain distribution via adsorptive-mediated transcytosis of an amyloid-beta (A beta) protofibril selective F(ab')2 fragment (F(ab')2-h158). F(ab')2-h158 was cationized to d...

  7. Tanyproctini (Coleoptera: Scarabaeidae: Melolonthinae) of Socotra Island

    Czech Academy of Sciences Publication Activity Database

    Král, D.; Sehnal, R.; Bezděk, Aleš

    2012-01-01

    Roč. 52, suppl. 2 (2012), s. 153-182 ISSN 0374-1036 Grant - others:Ministry of Education, Youth and Sports (CZ) LA10036/MSMT Institutional support: RVO:60077344 Keywords : Coleoptera * Scarabaeidae * Melolonthinae Subject RIV: EG - Zoology Impact factor: 0.963, year: 2012 http://www.aemnp.eu/PDF/52_s2/52_S2_153.pdf

  8. Species delineation and hybrid identification using diagnostic nuclear markers for Plectropomus leopardus and Plectropomus maculatus

    KAUST Repository

    He, Song

    2018-06-01

    Diagnostic molecular markers are an essential tool in the study of species’ ecology and evolution, particularly in closely related and sympatric species. Furthermore, the increasing awareness of wild-hybrids has led to a renewed interest in rapid diagnostic assays. Here, we test the ability of two mitochondrial (Cytb and COI) and two nuclear markers (ETS2 and TMO-4c4) to confidently discriminate purebred P. leopardus and P. maculatus and their first-generation hybrids. A sample of 48 purebred individuals and 91 interspecific hybrids were used in this study and their delineation confirmed using a set of microsatellite markers. Our results indicate mitochondrial markers could not distinguish even between species but both nuclear markers confidently identified species and first-generation hybrids. However, later-generation hybrids could not always be confidently identified due to on-going introgression between species. Our findings provide a robust tool to distinguish purebred individuals and interspecific hybrids in a pair of species with an unexpectedly high incidence of hybridization. The quick species discrimination abilities provided by these diagnostic markers are important for stock assessment and recruitment studies of these important fishery species.

  9. Lipid Status of the Two High Latitude Fish Species, Leptoclinus maculatus and Lumpenus fabricii

    Directory of Open Access Journals (Sweden)

    Nina N. Nemova

    2013-03-01

    Full Text Available A comparative study of the lipid status (i.e., the total lipid and phospholipid concentrations and the percentage of fatty acids of the total lipids of adult specimens of daubed shanny (Leptoclinus maculatus from Svalbard waters (Isfjord and slender eel blenny (Lumpenus fabricii from the White Sea (Onega Bay and Tersky shore was performed to study the metabolism and functions of lipids of these fishes in ontogeny and under various ecological conditions. Slender eel blenny from both areas of the White Sea were distinguished by a high level of sphingomyelin compared with the daubed shanny from Svalbard, and the amount of total phospholipids was higher in slender eel blenny from Onega Bay than in slender eel blenny from the Tersky shore. The extent of saturation and the signature of polyenic fatty acids varied according to the specific species of the Stichaeidae family under study. These results demonstrate the differences in the trophoecological and hydrobiological conditions of habitations of these species and highlighted the importance of considering certain trends in the lipid profiles of these fishes as specific features of the organization of the ecological and biochemical mechanisms of adaptation.

  10. Phylogeography in Galaxias maculatus (Jenyns, 1848) along Two Biogeographical Provinces in the Chilean Coast

    Science.gov (United States)

    González-Wevar, Claudio A.; Salinas, Pilar; Hüne, Mathias; Segovia, Nicolás I.; Vargas-Chacoff, Luis; Astorga, Marcela; Cañete, Juan I.; Poulin, Elie

    2015-01-01

    Major geologic and climatic changes during the Quaternary exerted a major role in shaping past and contemporary distribution of genetic diversity and structure of aquatic organisms in southern South America. In fact, the northern glacial limit along the Pacific coast, an area of major environmental changes in terms of topography, currents, and water salinity, represents a major biogeographic transition for marine and freshwater species. We used mitochondrial DNA sequences (D-loop) to investigate the consequences of Quaternary glacial cycles over the pattern of genetic diversity and structure of G. maculatus (Pisces: Galaxiidae) along two biogeographical provinces in the Chilean coast. Extreme levels of genetic diversity and strong phylogeographic structure characterize the species suggesting a low amount of influence of the last glacial cycle over its demography. However, we recognized contrasting patterns of genetic diversity and structure between main biogeographical areas here analyzed. Along the Intermediate Area (38°–41° S) each estuarine population constitutes a different unit. In contrast, Magellanic populations (43°–53° S) exhibited low levels of genetic differentiation. Contrasting patterns of genetic diversity and structure recorded in the species between the analyzed biogeographic areas are consistent with the marked differences in abiotic factors (i.e., different coastal configurations, Quaternary glacial histories, and oceanographic regimes) and to inherent characteristics of the species (i.e., salt-tolerance, physiology, and reproductive behavior). PMID:26161896

  11. Salinity-dependent mechanisms of copper toxicity in the galaxiid fish, Galaxias maculatus.

    Science.gov (United States)

    Glover, Chris N; Urbina, Mauricio A; Harley, Rachel A; Lee, Jacqueline A

    2016-05-01

    The euryhaline galaxiid fish, inanga (Galaxias maculatus) is widely spread throughout the Southern hemisphere occupying near-coastal streams that may be elevated in trace elements such as copper (Cu). Despite this, nothing is known regarding their sensitivity to Cu contamination. The mechanisms of Cu toxicity in inanga, and the ameliorating role of salinity, were investigated by acclimating fish to freshwater (FW), 50% seawater (SW), or 100% SW and exposing them to a graded series of Cu concentrations (0-200μgL(-1)) for 48h. Mortality, whole body Cu accumulation, measures of ionoregulatory disturbance (whole body ions, sodium (Na) influx, sodium/potassium ATPase activity) and ammonia excretion were monitored. Toxicity of Cu was greatest in FW, with mortality likely resulting from impaired Na influx. In both FW and 100% SW, ammonia excretion was significantly elevated, an effect opposite to that observed in previous studies, suggesting fundamental differences in the effect of Cu in this species relative to other studied fish. Salinity was protective against Cu toxicity, and physiology seemed to play a more important role than water chemistry in this protection. Inanga are sensitive to waterborne Cu through a conserved impairment of Na ion homeostasis, but some effects of Cu exposure in this species are distinct. Based on effect concentrations, current regulatory tools and limits are likely protective of this species in New Zealand waters. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Phylogeography in Galaxias maculatus (Jenyns, 1848 along Two Biogeographical Provinces in the Chilean Coast.

    Directory of Open Access Journals (Sweden)

    Claudio A González-Wevar

    Full Text Available Major geologic and climatic changes during the Quaternary exerted a major role in shaping past and contemporary distribution of genetic diversity and structure of aquatic organisms in southern South America. In fact, the northern glacial limit along the Pacific coast, an area of major environmental changes in terms of topography, currents, and water salinity, represents a major biogeographic transition for marine and freshwater species. We used mitochondrial DNA sequences (D-loop to investigate the consequences of Quaternary glacial cycles over the pattern of genetic diversity and structure of G. maculatus (Pisces: Galaxiidae along two biogeographical provinces in the Chilean coast. Extreme levels of genetic diversity and strong phylogeographic structure characterize the species suggesting a low amount of influence of the last glacial cycle over its demography. However, we recognized contrasting patterns of genetic diversity and structure between main biogeographical areas here analyzed. Along the Intermediate Area (38°-41° S each estuarine population constitutes a different unit. In contrast, Magellanic populations (43°-53° S exhibited low levels of genetic differentiation. Contrasting patterns of genetic diversity and structure recorded in the species between the analyzed biogeographic areas are consistent with the marked differences in abiotic factors (i.e., different coastal configurations, Quaternary glacial histories, and oceanographic regimes and to inherent characteristics of the species (i.e., salt-tolerance, physiology, and reproductive behavior.

  13. Effects of dietary antioxidant of tomato extract and lycopene on Carassius auratus and Xiphophorus maculatus

    Directory of Open Access Journals (Sweden)

    Cynthia Montoya M.

    2014-06-01

    Full Text Available Objective. Evaluate the effect on tegument pigmentation, survival, growth and antioxidant capacity in diets supplemented with tomato extract and lycopene as additives in experimental feed for Carassius auratus and Xiphophorus maculatus. Materials and methods. The additives were added in different concentrations to a basic diet. We performed beginning and an ending biometrics for 100% of the population in each bioassay. The growth and survival of organisms were evaluated. The antioxidant capacity was analyzed by ABTS assay, both in the tomato extract sample as well as in foods used in different bioassays. The concentration of lycopene was determined in food and liver and muscle samples of fish fed with it. Acquired pigmentation of fish was assessed through photographs analyzed with Adobe Photoshop®. The results were evaluated by analysis of variance, and when differences were found (p0.05 on pigmentation and growth of the organisms under the established experimental conditions was obtained. Significant differences in antioxidant capacity (p<0.05 were obtained in foods with added lycopene. Conclusions. The inclusion of lycopene or tomato extract in food for the organisms used is not recommended to improve pigmentation, but further studies are needed to demonstrate antioxidant effect.

  14. Sexual competion of irradiated males of the cowpea weevil, callosobruchus maculatus (F.)

    International Nuclear Information System (INIS)

    Ahmed, M.Y.Y.; Younes, M.W.F.

    1981-01-01

    When unmated males of Callosobruchus maculatus were irradiated with 7.5 or 10 Krad and confined with normal females, 97.90 and 89.32%, respectively, of the eggs did not hatch, compared with 15.49% infertility of the control eggs. Increasing the dose to 12.5 or 15 Krad induced 99.44 and 100%, respectively, sterility in the irradiated (I) males. When males treated with 7.5, 10 or 12.5 Krad were confined with unirradiated (U) males and females (1:1:1 ratio), infertility of the resulting eggs were 60.06, 67.96 and 69.04%, respectively. The competitiveness values of irradiated males with the above doses and ratio were 1.2, 1.6 and 1.8. Thus, males treated with substerilizing doses (7.5, 10 or 12.5 Krad) competed favourably with (U) males, and males treated with the sterilizing dose (15 Krad) were also fully competitive with (U) males. These results indicated that irradiation with 15 Krad, a sterilizing dose, did not decrease sexual competitiveness of irradiated males. (author)

  15. Species delineation and hybrid identification using diagnostic nuclear markers for Plectropomus leopardus and Plectropomus maculatus

    KAUST Repository

    He, Song; Harrison, Hugo B.; Berumen, Michael L.

    2018-01-01

    Diagnostic molecular markers are an essential tool in the study of species’ ecology and evolution, particularly in closely related and sympatric species. Furthermore, the increasing awareness of wild-hybrids has led to a renewed interest in rapid diagnostic assays. Here, we test the ability of two mitochondrial (Cytb and COI) and two nuclear markers (ETS2 and TMO-4c4) to confidently discriminate purebred P. leopardus and P. maculatus and their first-generation hybrids. A sample of 48 purebred individuals and 91 interspecific hybrids were used in this study and their delineation confirmed using a set of microsatellite markers. Our results indicate mitochondrial markers could not distinguish even between species but both nuclear markers confidently identified species and first-generation hybrids. However, later-generation hybrids could not always be confidently identified due to on-going introgression between species. Our findings provide a robust tool to distinguish purebred individuals and interspecific hybrids in a pair of species with an unexpectedly high incidence of hybridization. The quick species discrimination abilities provided by these diagnostic markers are important for stock assessment and recruitment studies of these important fishery species.

  16. Automated reticle inspection data analysis for wafer fabs

    Science.gov (United States)

    Summers, Derek; Chen, Gong; Reese, Bryan; Hutchinson, Trent; Liesching, Marcus; Ying, Hai; Dover, Russell

    2009-04-01

    To minimize potential wafer yield loss due to mask defects, most wafer fabs implement some form of reticle inspection system to monitor photomask quality in high-volume wafer manufacturing environments. Traditionally, experienced operators review reticle defects found by an inspection tool and then manually classify each defect as 'pass, warn, or fail' based on its size and location. However, in the event reticle defects are suspected of causing repeating wafer defects on a completed wafer, potential defects on all associated reticles must be manually searched on a layer-by-layer basis in an effort to identify the reticle responsible for the wafer yield loss. This 'problem reticle' search process is a very tedious and time-consuming task and may cause extended manufacturing line-down situations. Often times, Process Engineers and other team members need to manually investigate several reticle inspection reports to determine if yield loss can be tied to a specific layer. Because of the very nature of this detailed work, calculation errors may occur resulting in an incorrect root cause analysis effort. These delays waste valuable resources that could be spent working on other more productive activities. This paper examines an automated software solution for converting KLA-Tencor reticle inspection defect maps into a format compatible with KLA-Tencor's Klarity Defect(R) data analysis database. The objective is to use the graphical charting capabilities of Klarity Defect to reveal a clearer understanding of defect trends for individual reticle layers or entire mask sets. Automated analysis features include reticle defect count trend analysis and potentially stacking reticle defect maps for signature analysis against wafer inspection defect data. Other possible benefits include optimizing reticle inspection sample plans in an effort to support "lean manufacturing" initiatives for wafer fabs.

  17. Reclassification of leukemia among A-bomb survivors by French-American-British (FAB) classification, 1

    International Nuclear Information System (INIS)

    Matsuo, Tatsuki; Tomonaga, Masao; Bennett, J.M.

    1987-09-01

    The concordance rate for the French-American-British (FAB) reclassification diagnoses of atomic bomb-related cases of leukemia in Nagasaki was determined by a group of RERF hematologists and one of the members of the FAB cooperative gruop. The peripheral blood and/or bone marrow smears from 193 persons with leukemia or related disorder were reviewed. There was 85% agreement in the identification of leukemia types and subtypes. There was almost complete agreement for the diagnosis of non-FAB disorders (chronic myeloid leukemia and others) resulting in overall concordance of 88.2%. The conclusion from this remarkably high rate of concordance is that it is feasible to accurately apply the FAB classification system to the cases of A-bomb-related leukemia. These preliminary observations suggest that the previously established leukemia types for about a quarter of the cases of acute leukemia and related disorders should be changed. (author)

  18. Genome-Wide SNP Discovery, Genotyping and Their Preliminary Applications for Population Genetic Inference in Spotted Sea Bass (Lateolabrax maculatus.

    Directory of Open Access Journals (Sweden)

    Juan Wang

    Full Text Available Next-generation sequencing and the collection of genome-wide single-nucleotide polymorphisms (SNPs allow identifying fine-scale population genetic structure and genomic regions under selection. The spotted sea bass (Lateolabrax maculatus is a non-model species of ecological and commercial importance and widely distributed in northwestern Pacific. A total of 22 648 SNPs was discovered across the genome of L. maculatus by paired-end sequencing of restriction-site associated DNA (RAD-PE for 30 individuals from two populations. The nucleotide diversity (π for each population was 0.0028±0.0001 in Dandong and 0.0018±0.0001 in Beihai, respectively. Shallow but significant genetic differentiation was detected between the two populations analyzed by using both the whole data set (FST = 0.0550, P < 0.001 and the putatively neutral SNPs (FST = 0.0347, P < 0.001. However, the two populations were highly differentiated based on the putatively adaptive SNPs (FST = 0.6929, P < 0.001. Moreover, a total of 356 SNPs representing 298 unique loci were detected as outliers putatively under divergent selection by FST-based outlier tests as implemented in BAYESCAN and LOSITAN. Functional annotation of the contigs containing putatively adaptive SNPs yielded hits for 22 of 55 (40% significant BLASTX matches. Candidate genes for local selection constituted a wide array of functions, including binding, catalytic and metabolic activities, etc. The analyses with the SNPs developed in the present study highlighted the importance of genome-wide genetic variation for inference of population structure and local adaptation in L. maculatus.

  19. Morphological and molecular data reveal a new species of Neoechinorhynchus (Acanthocephala: Neoechinorhynchidae) from Dormitator maculatus in the Gulf of Mexico.

    Science.gov (United States)

    Pinacho-Pinacho, Carlos Daniel; Sereno-Uribe, Ana L; García-Varela, Martín

    2014-12-01

    Neoechinorhynchus (Neoechinorhynchus) mexicoensis sp. n. is described from the intestine of Dormitator maculatus (Bloch 1792) collected in 5 coastal localities from the Gulf of Mexico. The new species is mainly distinguished from the other 33 described species of Neoechinorhynchus from the Americas associated with freshwater, marine and brackish fishes by having smaller middle and posterior hooks and possessing a small proboscis with three rows of six hooks each, apical hooks longer than other hooks and extending to the same level as the posterior hooks, 1 giant nucleus in the ventral body wall and females with eggs longer than other congeneric species. Sequences of the internal transcribed spacer (ITS) and the large subunit (LSU) of ribosomal DNA including the domain D2+D3 were used independently to corroborate the morphological distinction among the new species and other congeneric species associated with freshwater and brackish water fish from Mexico. The genetic divergence estimated among congeneric species ranged from 7.34 to 44% for ITS and from 1.65 to 32.9% for LSU. Maximum likelihood and Bayesian inference analyses with each dataset showed that the 25 specimens analyzed from 5 localities of the coast of the Gulf of Mexico parasitizing D. maculatus represent an independent clade with strong bootstrap support and posterior probabilities. The morphological evidence, plus the monophyly in the phylogenetic analyses, indicates that the acanthocephalans collected from intestine of D. maculatus from the Gulf of Mexico represent a new species, herein named N. (N.) mexicoensis sp. n. Copyright © 2014. Published by Elsevier Ireland Ltd.

  20. Comparative imaging and biodistribution studies with an anti-CEA monoclonal antibody and its F(ab)2 and Fab fragments in mice with colon carcinoma xenografts

    International Nuclear Information System (INIS)

    Andrew, S.M.; Pimm, M.V.; Baldwin, R.W.; Perkins, A.C.

    1986-01-01

    An IgG1 mouse monoclonal antibody directed against CEA has been digested with papain to yield F(ab) 2 and Fab fragments. Following radioiodination, intact antibody and fragments showed specific binding to cells of a CEA-producing tumour, although the immune reactivities of the fragments were lower than that of intact antibody. Gamma scintigraphy of nude mice bearing CEA producing human tumour xenografts and injected with 131 I-labelled fragments showed earlier and superior imaging of tumours than did 131 I-intact antibody, and this was most marked with the Fab fragment. Sequential dissection analyses showed that this was due to earlier and higher tumour-to-blood ratios with fragments than with intact antibody, but in absolute terms the degree of localization of both fragment types was significantly lower than that of intact antibody. (orig.)

  1. Near-Atomic Resolution Structure of a Highly Neutralizing Fab Bound to Canine Parvovirus.

    Science.gov (United States)

    Organtini, Lindsey J; Lee, Hyunwook; Iketani, Sho; Huang, Kai; Ashley, Robert E; Makhov, Alexander M; Conway, James F; Parrish, Colin R; Hafenstein, Susan

    2016-11-01

    Canine parvovirus (CPV) is a highly contagious pathogen that causes severe disease in dogs and wildlife. Previously, a panel of neutralizing monoclonal antibodies (MAb) raised against CPV was characterized. An antibody fragment (Fab) of MAb E was found to neutralize the virus at low molar ratios. Using recent advances in cryo-electron microscopy (cryo-EM), we determined the structure of CPV in complex with Fab E to 4.1 Å resolution, which allowed de novo building of the Fab structure. The footprint identified was significantly different from the footprint obtained previously from models fitted into lower-resolution maps. Using single-chain variable fragments, we tested antibody residues that control capsid binding. The near-atomic structure also revealed that Fab binding had caused capsid destabilization in regions containing key residues conferring receptor binding and tropism, which suggests a mechanism for efficient virus neutralization by antibody. Furthermore, a general technical approach to solving the structures of small molecules is demonstrated, as binding the Fab to the capsid allowed us to determine the 50-kDa Fab structure by cryo-EM. Using cryo-electron microscopy and new direct electron detector technology, we have solved the 4 Å resolution structure of a Fab molecule bound to a picornavirus capsid. The Fab induced conformational changes in regions of the virus capsid that control receptor binding. The antibody footprint is markedly different from the previous one identified by using a 12 Å structure. This work emphasizes the need for a high-resolution structure to guide mutational analysis and cautions against relying on older low-resolution structures even though they were interpreted with the best methodology available at the time. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  2. Investigation of protein selectivity in multimodal chromatography using in silico designed Fab fragment variants.

    Science.gov (United States)

    Karkov, Hanne Sophie; Krogh, Berit Olsen; Woo, James; Parimal, Siddharth; Ahmadian, Haleh; Cramer, Steven M

    2015-11-01

    In this study, a unique set of antibody Fab fragments was designed in silico and produced to examine the relationship between protein surface properties and selectivity in multimodal chromatographic systems. We hypothesized that multimodal ligands containing both hydrophobic and charged moieties would interact strongly with protein surface regions where charged groups and hydrophobic patches were in close spatial proximity. Protein surface property characterization tools were employed to identify the potential multimodal ligand binding regions on the Fab fragment of a humanized antibody and to evaluate the impact of mutations on surface charge and hydrophobicity. Twenty Fab variants were generated by site-directed mutagenesis, recombinant expression, and affinity purification. Column gradient experiments were carried out with the Fab variants in multimodal, cation-exchange, and hydrophobic interaction chromatographic systems. The results clearly indicated that selectivity in the multimodal system was different from the other chromatographic modes examined. Column retention data for the reduced charge Fab variants identified a binding site comprising light chain CDR1 as the main electrostatic interaction site for the multimodal and cation-exchange ligands. Furthermore, the multimodal ligand binding was enhanced by additional hydrophobic contributions as evident from the results obtained with hydrophobic Fab variants. The use of in silico protein surface property analyses combined with molecular biology techniques, protein expression, and chromatographic evaluations represents a previously undescribed and powerful approach for investigating multimodal selectivity with complex biomolecules. © 2015 Wiley Periodicals, Inc.

  3. A universal phage display system for the seamless construction of Fab libraries.

    Science.gov (United States)

    Nelson, Renae S; Valadon, Philippe

    2017-11-01

    The construction of Fab phage libraries requires the cloning of domains from both the light and the heavy chain of antibodies. Despite the advent of powerful strategies such as splicing-by-overlap extension PCR, obtaining high quality libraries with excellent coverage remains challenging. Here, we explored the use of type IIS restriction enzymes for the seamless cloning of Fab libraries. We analyzed human, murine and rabbit germline antibody repertoires and identified combinations of restriction enzymes that exhibit very few or no recognition sites in the antibody sequences. We describe three phagemid vectors, pUP-22Hb, pUP-22Mc and pUP-22Rc, which were employed for cloning the Fab repertoire of these hosts using BsmBI and SapI (human) or SapI alone (mouse and rabbit). Using human serum albumin as a model immunization, we built a mouse/human chimeric Fab library and a mouse Fab library in a single step ligation and successfully panned multiple cognate antibodies. The overall process is highly scalable and faster than PCR-based techniques, with a Fab insertion success rate of around 80%. By using carefully chosen overhangs on each end of the antibody domains, this approach paves the way to the universal, sequence- and vector-independent cloning and reformatting of antibody libraries. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Fab is the most efficient format to express functional antibodies by yeast surface display.

    Science.gov (United States)

    Sivelle, Coline; Sierocki, Raphaël; Ferreira-Pinto, Kelly; Simon, Stéphanie; Maillere, Bernard; Nozach, Hervé

    2018-04-30

    Multiple formats are available for engineering of monoclonal antibodies (mAbs) by yeast surface display, but they do not all lead to efficient expression of functional molecules. We therefore expressed four anti-tumor necrosis factor and two anti-IpaD mAbs as single-chain variable fragment (scFv), antigen-binding fragment (Fab) or single-chain Fabs and compared their expression levels and antigen-binding efficiency. Although the scFv and scFab formats are widely used in the literature, 2 of 6 antibodies were either not or weakly expressed. In contrast, all 6 antibodies expressed as Fab revealed strong binding and high affinity, comparable to that of the soluble form. We also demonstrated that the variations in expression did not affect Fab functionality and were due to variations in light chain display and not to misfolded dimers. Our results suggest that Fab is the most versatile format for the engineering of mAbs.

  5. Effect of DNA sequence of Fab fragment on yield characteristics and cell growth of E. coli.

    Science.gov (United States)

    Kulmala, Antti; Huovinen, Tuomas; Lamminmäki, Urpo

    2017-06-19

    Codon usage is one of the factors influencing recombinant protein expression. We were interested in the codon usage of an antibody Fab fragment gene exhibiting extreme toxicity in the E. coli host. The toxic synthetic human Fab gene contained domains optimized by the "one amino acid-one codon" method. We redesigned five segments of the Fab gene with a "codon harmonization" method described by Angov et al. and studied the effects of these changes on cell viability, Fab yield and display on filamentous phage using different vectors and bacterial strains. The harmonization considerably reduced toxicity, increased Fab expression from negligible levels to 10 mg/l, and restored the display on phage. Testing the impact of the individual redesigned segments revealed that the most significant effects were conferred by changes in the constant domain of the light chain. For some of the Fab gene variants, we also observed striking differences in protein yields when cloned from a chloramphenicol resistant vector into an identical vector, except with ampicillin resistance. In conclusion, our results show that the expression of a heterodimeric secretory protein can be improved by harmonizing selected DNA segments by synonymous codons and reveal additional complexity involved in heterologous protein expression.

  6. Functional Requirements for Fab-7 Boundary Activity in the Bithorax Complex

    Science.gov (United States)

    Wolle, Daniel; Cleard, Fabienne; Aoki, Tsutomu; Deshpande, Girish; Karch, Francois

    2015-01-01

    Chromatin boundaries are architectural elements that determine the three-dimensional folding of the chromatin fiber and organize the chromosome into independent units of genetic activity. The Fab-7 boundary from the Drosophila bithorax complex (BX-C) is required for the parasegment-specific expression of the Abd-B gene. We have used a replacement strategy to identify sequences that are necessary and sufficient for Fab-7 boundary function in the BX-C. Fab-7 boundary activity is known to depend on factors that are stage specific, and we describe a novel ∼700-kDa complex, the late boundary complex (LBC), that binds to Fab-7 sequences that have insulator functions in late embryos and adults. We show that the LBC is enriched in nuclear extracts from late, but not early, embryos and that it contains three insulator proteins, GAF, Mod(mdg4), and E(y)2. Its DNA binding properties are unusual in that it requires a minimal sequence of >65 bp; however, other than a GAGA motif, the three Fab-7 LBC recognition elements display few sequence similarities. Finally, we show that mutations which abrogate LBC binding in vitro inactivate the Fab-7 boundary in the BX-C. PMID:26303531

  7. Production and characterization of anti-human IgG F(ab')2 antibody fragment.

    Science.gov (United States)

    Valedkarimi, Zahra; Nasiri, Hadi; Aghebati-Maleki, Leili; Abdolalizadeh, Jalal; Esparvarinha, Mojghan; Majidi, Jafar

    2018-04-10

    In present study an optimized protocol for the separation of antibodies into antigen-binding fragments F(ab')2 using pepsin digestion was investigated. The production of these fragments is a consequential step in the development of medical research, treatment and diagnosis. For production of polyclonal antibody rabbit received antigen in four steps. The rabbit serum at 1/128000 dilution showed high absorbance in reaction with human IgG at the designed ELISA method. Rabbit IgG was purified by Ion-Exchange Chromatography (IEC) method. Purity was assessed by SDS-PAGE method. In non-reduced condition only one band was seen in about 150 kDa MW position and in reduced form, two bands were seen in 50 and 25 kDa MW positions. Rabbit IgG was digested by pepsin enzyme. The antibody fragments solution was applied to Gel filtration column to isolate the F(ab')2. Non-reduced SDS-PAGE for determining the purity of F(ab')2 fragment resulted in one band in 100 kDa corresponds to F(ab')2 fragment and a band in 150 kDa MW position corresponds to undigested IgG antibodies. The activities of FITC conjugated F(ab')2 fragment and commercial ones were compared using flowcytometry method. The activity results implied that the FITC conjugated- anti human F(ab')2 fragment worked as efficiently as the commercial one.

  8. Functional Requirements for Fab-7 Boundary Activity in the Bithorax Complex.

    Science.gov (United States)

    Wolle, Daniel; Cleard, Fabienne; Aoki, Tsutomu; Deshpande, Girish; Schedl, Paul; Karch, Francois

    2015-11-01

    Chromatin boundaries are architectural elements that determine the three-dimensional folding of the chromatin fiber and organize the chromosome into independent units of genetic activity. The Fab-7 boundary from the Drosophila bithorax complex (BX-C) is required for the parasegment-specific expression of the Abd-B gene. We have used a replacement strategy to identify sequences that are necessary and sufficient for Fab-7 boundary function in the BX-C. Fab-7 boundary activity is known to depend on factors that are stage specific, and we describe a novel ∼700-kDa complex, the late boundary complex (LBC), that binds to Fab-7 sequences that have insulator functions in late embryos and adults. We show that the LBC is enriched in nuclear extracts from late, but not early, embryos and that it contains three insulator proteins, GAF, Mod(mdg4), and E(y)2. Its DNA binding properties are unusual in that it requires a minimal sequence of >65 bp; however, other than a GAGA motif, the three Fab-7 LBC recognition elements display few sequence similarities. Finally, we show that mutations which abrogate LBC binding in vitro inactivate the Fab-7 boundary in the BX-C. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. Characterization of Human Colorectal Cancer MDR1/P-gp Fab Antibody

    Directory of Open Access Journals (Sweden)

    Xuemei Zhang

    2013-01-01

    Full Text Available In this study, the peptide sized 21 kDa covering P-gp transmembrane region was first prepared for generating a novel mouse monoclonal antibody Fab fragment with biological activity against multiple drug resistance protein P-gp21 by phage display technology. Phage-displayed antibody library prepared from mice spleen tissues was selected against the recombinant protein P-gp21 with five rounds of panning. A number of clones expressing Fab bound to P-gp21, showing neutralized activity in vitro, were isolated and screened by enzyme-linked immunosorbent assay based on its recognition properties to P-gp21 and human colorectal cancer tissue homogenate, resulting in identification of an optimal recombinant Fab clone (Number 29. Further characterization by recloning number 29 into an expression vector showed significant induction of the Fab antibody in the clone number 29 by Isopropyl β-D-1-thiogalactopyranoside (IPTG. After purified by HiTrap Protein L, the specificity of the Fab antibody to P-gp21 was also confirmed. Not only was the targeted region of this monoclonal Fab antibody identified as a 16-peptide epitope (ALKDKKELEGSGKIAT comprising residues 883–898 within the transmembrane (TM domain of human P-gp, but also the binding ability with it was verified. The clinical implication of our results for development of personalized therapy of colorectal cancer will be further studied.

  10. Use of I-131 labeled, murine Fab against a high molecular weight antigen of human melanoma: Preliminary experience

    International Nuclear Information System (INIS)

    Larson, S.M.; Carrasquillo, J.A.; McGuffin, R.W.

    1985-01-01

    High molecular-weight antigen (HMWA) is tumor-associated proteoglycan of human malignant melanoma. I-131 labeled Fab fragments of these specific antibodies were used for preliminary feasibility studies for radioimmunodetection and therapy of human subjects who had inoperable metastatic melanoma. Ten patients received tracer doses of I-131 (anti-HMWA) Fab. All patients (8/8) who had melanoma lesions greater than 1 cm by correlative diagnosis methods had one or more lesions that had localization to tumor of the radiolabelled Fab. In all, 17 of 23 (74%) documented metastases were seen. Two patients who had avid uptake received potentially radiotherapeutic doses. For both of these patients, whole imaging studies showed that the localization of the high dose I-131 Fab was predominantly in tumor. On whole body images, the anti-Fab HMWA appears to be more tumor selective than Fab preparations that target the p97 antigen for melanoma, and there is less uptake in liver

  11. Efecto de la temperatura en la sobrevivencia embrionaria del puye Galaxias maculatus (Jenyns, 1842

    Directory of Open Access Journals (Sweden)

    Juan Bariles

    2013-11-01

    Full Text Available Se evaluó el efecto de seis temperaturas constantes en la sobrevivencia embrionaria, periodo embrionario y periodo de eclosión de Galaxias maculatus de 7 a 22°C, en condiciones de laboratorio. Los resultados de la sobrevivencia embrionaria fueron: 90,6 ± 2,0; 88,7 ± 8,6; 87,7 ± 0,7; 89,7 ± 4,2; 70,0 ± 5,6 y 49,5 ± 7,4% a temperaturas de 7, 10, 13, 16, 19 y 22°C. Exceptuando la temperatura de 22°C, no hay diferencias significativas entre ellas (P > 0,05. Estos resultados caracterizan al embrión como euritérmico independientemente de ser una especie de aguas subantárticas. En relación al periodo embrionario (tH50%, los resultados fueron de 40,8 ± 0,6; 27,0 ± 0,0; 17,3 ± 0,6; 16,3 ± 0,6 y 13,3 ± 0,6 días a temperaturas de 7, 10, 13, 16 y 19°C respectivamente, con diferencias significativas entre ellas (P 87,7%, periodos embrionarios <27 días y periodos de eclosión <6,3 días. Estos resultados permitirán optimizar la sobrevivencia embrionaria y el eventual cultivo de la especie para fines comerciales o de repoblamiento.

  12. Efecto de la salinidad en la supervivencia embrionaria de puye Galaxias maculatus (Jenyns, 1842

    Directory of Open Access Journals (Sweden)

    Juan Barile

    2015-05-01

    Full Text Available Se evaluó e l efecto de la salinidad en la supervivencia embrionaria, período embrionario y período de eclosión de puye, Galaxias maculatus. Se utilizaron ocho tratamientos, con salinidades de 0, 4, 8, 12, 16 y 20 obteniéndose supervivencias embrionarias promedios respectivas de 84,9; 84,0; 85,5; 86,1; 82,6 y 75,8%, sin diferencias significativas entre ellas, mientras que con salinidad de 24 se obtuvo sobrevivencia de 10,4%, significativamente diferente al resto de los tratamientos y con salinidad de 28 causó 100% de mortalidad. A diferencia del estado juvenil y adulto, el embrión es incapaz de resistir la salinidad marina y su umbral de tolerancia se encuentra entre salinidades de 24 y 28. Las mayores supervivencias promedios larvales a 10 días de vida fueron con salinidades intermedias de 8, 12 y 16 de 79,3; 80,3 y 74,6% respectivamente, sin diferencias significativas entre ellas, demostrando alta viabilidad del embrión post-eclosión. Los períodos embrionarios promedios (50% más breves fueron a bajas salinidades 0, 4, 8, 12 con 28,7; 28,0; 29,7 y 29,7 días y los más extensos en los tratamientos con salinidades de 16 y 20, con 34,0 y 34,3 días, sin diferencias significativas entre ellos. Los períodos de eclosión promedio (50% con salinidades de 0, 4, 8, 12, 16, 20 y 24 fueron de 5,0; 5,5; 7,5; 8,8; 13,0; 14,3 y 15,8 días respectivamente, sin diferencias significativas entre ellos.

  13. New species of Elaphidiini (Coleoptera, Cerambycidae) from Bolivia.

    Science.gov (United States)

    Galileo, Maria Helena M; Martins, Ubirajara R; Santos-Silva, Antonio

    2014-11-14

    Five new Elaphidiini species are described from Bolivia including Anelaphus maculatus sp. nov. from Tarija and Mephritus bonasoi sp. nov., M. meyeri, sp. nov., M. punctulatus sp. nov. and M. eleandroi sp. nov. from Santa Cruz. A revised key to all 22 South American Mephritus species is provided.

  14. Effect of Gamma Radiation on Morphology and Histology of Sex Pheromone Gland of Callosobruchus maculatus (F.) Female

    International Nuclear Information System (INIS)

    El-Degwi, M.S.; Salem, H.M.; El-Maasarawy, S.A.S.; Ismail, I.I.

    2008-01-01

    The pheromone gland in normal adult female of C. maculatus consists of an internal lobe composed of glandular epithelia cells, connected dorsally with the abdominal tip. It has many tubule opening dorsally in large sets which have a groove. The size of pheromone gland of 4-day old females increased than 2-day old females. There is no difference between the gland size of 6-day old females and 4-day ones, but it decreased in females of 8-day old. In this age the gland divided into two separate parts. When females 4-day old irradiated with 100 Gy the glandular cells were separated and the nuclei were not clear. The fat bodies were less in numbers than in unirradiated ones and the glandular cell membrane is irregular. After increasing the radiation dose to 200 Gy, the glandular cells destroyed and become undistinguished. All the tubule were closed. The glandular cells in the irradiated 6-day old females with 100 Gy were compact. The nuclei and cytoplasm are very difficult to distinguished, the cytoplasm is deteriorated. As increasing the irradiation dose to 200 Gy, the glandular tissue completely damaged and appeared as narrow ribbon. Scanning electron micrographs of the abdominal tip of C.maculatus showed that the sex pheromone releasing area contains a mammiform, also it has a large number of large setae which has groove

  15. Essential oil optimizes the susceptibility of Callosobruchus maculatus and enhances the nutritional qualities of stored cowpea Vigna unguiculata.

    Science.gov (United States)

    Akami, Mazarin; Chakira, Hamada; Andongma, Awawing A; Khaeso, Kanjana; Gbaye, Olajire A; Nicolas, Njintang Y; Nukenine, E-N; Niu, Chang-Ying

    2017-08-01

    The intensive use of synthetic pesticides in cowpea storage has led to the development of resistance by Callosobruchus maculatus and subsequent degradation of grain quality. In an attempt to circumvent these constraints, the susceptibility of C. maculatus to 2,2-dichlorovinyldimethyl phosphate (DDVP) and Lippia adoensis essential oil (EO) was investigated and variations in the proportions of nutritional values of treated grains 150 days after storage were assessed. The survival rate was recorded after five generations. The resistance index and biochemical parameters of grains were determined. The results from this study revealed that the survival rate and resistance index significantly increased proportionally with damage in DDVP treatments ( r  = 0.889; p  = 0.018) while in EO treatments, those values remained low without significant variations ( p  = 0.0764) throughout the generations. DDVP stored grains yielded higher crude protein values, but lower carbohydrates, tannins, phenolics and minerals compared to EO. Eighteen amino acids were detected in EO treated grains and 14 in DDVP which was devoid of albumin and prolamin. Lippia adoensis EO could therefore represent a safe alternative bio-pesticide to cope with insect resistance and enhance the nutritional qualities of stored cowpea seeds.

  16. Conversion of a Mouse Fab into a Whole Humanized IgG Antibody for Detecting Botulinum Toxin

    National Research Council Canada - National Science Library

    Palys, Thomas J; Schmid, Kara E; Scherer, John M; Schoepp, Randal J

    2006-01-01

    Antibodies serve as the gold standard in most immunodiagnostic assays. Recent advances in recombinant DNA technology have offered the production of antibody fragments or Fabs as promising alternatives...

  17. Phage Fab Display Selection In Vitro and In Vivo: Novel Means to Identify New Breast Cancer Avid Compounds

    National Research Council Canada - National Science Library

    Meighan, Mark

    2001-01-01

    .... In this annual report we present preliminary results on the isolation of antibody fragments (Fabs), isolated from phage display libraries, when affinity selected against breast cancer cell lines...

  18. Effect of polyethylene glycol conjugation on conformational and colloidal stability of a monoclonal antibody antigen-binding fragment (Fab').

    Science.gov (United States)

    Roque, Cristopher; Sheung, Anthony; Rahman, Nausheen; Ausar, S Fernando

    2015-02-02

    We have investigated the effects of site specific "hinge" polyethylene glycol conjugation (PEGylation) on thermal, pH, and colloidal stability of a monoclonal antibody antigen-binding fragment (Fab') using a variety of biophysical techniques. The results obtained by circular dichroism (CD), ultraviolet (UV) absorbance, and fluorescence spectroscopy suggested that the physical stability of the Fab' is maximized at pH 6-7 with no apparent differences due to PEGylation. Temperature-induced aggregation experiments revealed that PEGylation was able to increase the transition temperature, as well as prevent the formation of visible and subvisible aggregates. Statistical comparison of the three-index empirical phase diagram (EPD) revealed significant differences in thermal and pH stability signatures between Fab' and PEG-Fab'. Upon mechanical stress, micro-flow imaging (MFI) and measurement of the optical density at 360 nm showed that the PEG-Fab' had significantly higher resistance to surface-induced aggregation compared to the Fab'. Analysis of the interaction parameter, kD, indicated repulsive intermolecular forces for PEG-Fab' and attractive forces for Fab'. In conclusion, PEGylation appears to protect Fab' against thermal and mechanical stress-induced aggregation, likely due to a steric hindrance mechanism.

  19. Heavy metals in carabids (Coleoptera, Carabidae

    Directory of Open Access Journals (Sweden)

    Ruslan Butovsky

    2011-05-01

    Full Text Available Carabid beetles (Coleoptera, Carabidae are one of the most studied soil groups in relation to heavy metal (HM accumulation and use for bioindication of environmental pollution. Accumulation of Zn and Cu in carabid beetles was species-, sex- and trophic group-specific. No differences were found in HM contents between omnivorous and carnivorous species. The use of carabid beetles as indicators of HM accumulation appears to be rather limited.

  20. Effects of Fab' fragments of specific egg yolk antibody (IgY-Fab') against Shewanella putrefaciens on the preservation of refrigerated turbot.

    Science.gov (United States)

    Zhang, Qian; Lin, Hong; Sui, Jianxin; Wang, Jingxue; Cao, Limin

    2015-01-01

    In our previous studies the specific egg yolk antibody (IgY) against Shewanella putrefaciens (one of the specific spoilage organisms for marine products during aerobic chilling storage) demonstrated significant activity to prolong the shelf life of refrigerated fish. The exploitation of the antigen-binding fragment plus the hinge region (IgY-Fab') is now considered a promising method for improving the efficiency of such natural antimicrobial agents. The antimicrobial activity of IgY-Fab' against S. putrefaciens was investigated using refrigerated turbot as samples. By microbial, chemical and sensory tests, it was shown to be able to effectively inhibit bacterial growth and prolong the shelf life of samples, with an efficiency evaluated significantly higher than that of whole IgY with the same molarity. The interaction between IgY agents and S. putrefaciens cells was also investigated, and the IgY-Fab' showed a much greater ability to damage cell membranes than the whole IgY. Compared to whole IgY with the same molarity, IgY-Fab' demonstrated higher and more durable antimicrobial efficiency. Such a result was assumed to be closely related to its structural properties (such as the much lower molecular weight), which may enhance its ability to influence physiological activities of antigen bacteria, especially the property or/and structure of cell membranes. © 2014 Society of Chemical Industry.

  1. Management of Tissue Loss After Agkistrodon Snakebite: Appropriate Use of Crotalidae-Fab Antivenin.

    Science.gov (United States)

    Larson, Kenneth W; Schaefer, Keith R; Austin, Cindy; Norton, Rhy; Finley, Phillip J

    2016-01-01

    Although initially created for the treatment of rattlesnake (genus: Crotalus) bites, Crotalidae-Fab antivenin is used to treat many different pit viper envenomations. However, the efficacy of Crotalidae-Fab in preventing tissue loss from copperhead (Agkistrodon contortrix) or cottonmouth (Agkistrodon piscivorus) snakebites remains unclear. Recent reports show that Agkistrodon-related bites rarely require treatment beyond simple observation and pain control. The purpose of this study was to examine the amount of tissue loss in patients who received Crotalidae-Fab compared with those who did not after an Agkistrodon bite. After institutional review board approval, a retrospective study was completed at a Level 1 trauma center. Between 2009 and 2013, a total of 57 snakebites were identified. Of the 57 bites, the snake species was documented in 36 cases including 31 copperheads, 1 cottonmouth, and 4 rattlesnakes. The other 21 bites were from unknown or nonvenomous species. Of the 32 Agkistrodon-related bites, 15 patients received Crotalidae-Fab (average of 3 vials administered) and 17 did not receive Crotalidae-Fab. None of the 32 patients, regardless of treatment option, had tissue loss or required surgical interventions. Only 1 patient received Crotalidae-Fab and debridement of a vesicle associated with the bite. No clinically significant differences were observed between the groups. These findings support previous literature that failed to show added benefit of Crotalidae-Fab treatment for Agkistrodon bites beyond patient comfort and pain control. Evaluation of current protocols for Agkistrodon envenomations is warranted. Snakebite wound education in trauma physicians and nurses may decrease unnecessary use of antivenom medication.

  2. Fab Four: When John and George Play Gravitation and Cosmology

    Directory of Open Access Journals (Sweden)

    J.-P. Bruneton

    2012-01-01

    Full Text Available Scalar-tensor theories of gravitation attract again a great interest since the discovery of the Chameleon mechanism and of the Galileon models. The former allows reconciling the presence of a scalar field with the constraints from Solar System experiments. The latter leads to inflationary models that do not need ad hoc potentials. Further generalizations lead to a tensor-scalar theory, dubbed the “Fab Four,” with only first and second order derivatives of the fields in the equations of motion that self-tune to a vanishing cosmological constant. This model needs to be confronted with experimental data in order to constrain its large parameter space. We present some results regarding a subset of this theory named “John,” which corresponds to a nonminimal derivative coupling between the scalar field and the Einstein tensor in the action. We show that this coupling gives rise to an inflationary model with very unnatural initial conditions. Thus, we include the term named “George,” namely, a nonminimal, but nonderivative, coupling between the scalar field and Ricci scalar. We find a more natural inflationary model, and, by performing a post-Newtonian analysis, we derive the set of equations that constrain the parameter space with data from experiments in the Solar System.

  3. Coagulation parameters in copperhead compared to other Crotalinae envenomation: secondary analysis of the F(ab')2 versus Fab antivenom trial.

    Science.gov (United States)

    Gerardo, Charles J; Vissoci, Joao R Nickenig; Brown, Michael W J; Bush, Sean P

    2017-02-01

    Coagulation derangements in copperhead envenomation are considered less severe than other crotaline envenomations, resulting in recommendations to limit both coagulation testing and antivenom treatment. A prospective, blinded, multicenter, randomized clinical trial comparing the effectiveness of F(ab') 2 versus Fab antivenom in crotaline envenomation patients was completed in 2011. We determined the difference between coagulation parameters in copperhead compared to other crotaline envenomations. We performed a post hoc analysis comparing the coagulation parameters (platelets and fibrinogen) prospectively obtained in the aforementioned trial. All the patients received antivenom in one of three treatment arms [F(ab') 2 with maintenance, F(ab') 2 with placebo maintenance, or Fab with maintenance]. Coagulation parameters were measured at pretreatment baseline, during acute hospitalization, day 5, day 8, and day 15 post-envenomation. Mean platelet count and fibrinogen levels for the copperhead and other crotaline groups were compared. The platelet and fibrinogen point estimates with distribution are presented graphically over time. 122 patients were enrolled in the study. There were 22 patients with copperhead envenomation, 93 with other crotaline envenomations, and 7 that could not be definitively determined. The mean age was 42 (SD 20) years. There was a minor pretreatment difference in mean baseline platelet count between the copperhead group (246 × 109/L 95% CI 215, 277) compared to other crotaline envenomation patients (184 × 109/L 95% CI 167, 202). There was a modest pretreatment difference in mean fibrinogen level between copperhead patients (345 mg/dL 95% CI 277, 415) and other crotaline patients (261mg/dL 95% CI 241, 281). Pretreatment coagulation parameter means were normal and converged post treatment. On average, copperhead envenomations have less severe initial coagulation derangements. However, in mild envenomations, differences in laboratory

  4. Contribution of Antibody Hydrodynamic Size to Vitreal Clearance Revealed through Rabbit Studies Using a Species-Matched Fab.

    Science.gov (United States)

    Shatz, Whitney; Hass, Philip E; Mathieu, Mary; Kim, Hok Seon; Leach, Kim; Zhou, Michelle; Crawford, Yongping; Shen, Amy; Wang, Kathryn; Chang, Debby P; Maia, Mauricio; Crowell, Susan R; Dickmann, Leslie; Scheer, Justin M; Kelley, Robert F

    2016-09-06

    We have developed a tool Fab fragment of a rabbit monoclonal antibody that is useful for early evaluation in rabbit models of technologies for long acting delivery (LAD) of proteins to the eye. Using this Fab we show that vitreal clearance can be slowed through increased hydrodynamic size. Fab (G10rabFab) and Fab' (G10rabFab') fragments of a rabbit monoclonal antibody (G10rabIgG) were expressed in Chinese hamster ovary (CHO) cells and purified using antigen-based affinity chromatography. G10rabFab retains antigen-binding upon thermal stress (37 °C) for 8 weeks in phosphate-buffered saline (PBS) and can be detected in rabbit tissues using an antigen-based ELISA. Hydrodynamic radius, measured using quasi-elastic light scattering (QELS), was increased through site-specific modification of the G10rabFab' free cysteine with linear methoxy-polyethylene glycol(PEG)-maleimide of 20000 or 40000 molecular weight. Pharmacokinetic studies upon intravitreal dosing in New Zealand white rabbits were conducted on the G10rabFab and PEGylated G10rabFab'. Results of single and multidose pharmacokinetic experiments yield reproducible results and a vitreal half-life for G10rabFab of 3.2 days. Clearance from the eye is slowed through increased hydrodynamic size, with vitreal half-life showing a linear dependence on hydrodynamic radius (RH). A linear dependence of vitreal half-life on RH suggests that molecule diffusivity makes an important contribution to vitreal clearance. A method for prediction of vitreal half-life from RH measurements is proposed.

  5. Structural Characterisation of FabG from Yersinia pestis, a Key Component of Bacterial Fatty Acid Synthesis.

    Directory of Open Access Journals (Sweden)

    Jeffrey D Nanson

    Full Text Available Ketoacyl-acyl carrier protein reductases (FabG are ubiquitously expressed enzymes that catalyse the reduction of acyl carrier protein (ACP linked thioesters within the bacterial type II fatty acid synthesis (FASII pathway. The products of these enzymes, saturated and unsaturated fatty acids, are essential components of the bacterial cell envelope. The FASII reductase enoyl-ACP reductase (FabI has been the focus of numerous drug discovery efforts, some of which have led to clinical trials, yet few studies have focused on FabG. Like FabI, FabG appears to be essential for survival in many bacteria, similarly indicating the potential of this enzyme as a drug target. FabG enzymes are members of the short-chain alcohol dehydrogenase/reductase (SDR family, and like other SDRs, exhibit highly conserved secondary and tertiary structures, and contain a number of conserved sequence motifs. Here we describe the crystal structures of FabG from Yersinia pestis (YpFabG, the causative agent of bubonic, pneumonic, and septicaemic plague, and three human pandemics. Y. pestis remains endemic in many parts of North America, South America, Southeast Asia, and Africa, and a threat to human health. YpFabG shares a high degree of structural similarity with bacterial homologues, and the ketoreductase domain of the mammalian fatty acid synthase from both Homo sapiens and Sus scrofa. Structural characterisation of YpFabG, and comparison with other bacterial FabGs and the mammalian fatty acid synthase, provides a strong platform for virtual screening of potential inhibitors, rational drug design, and the development of new antimicrobial agents to combat Y. pestis infections.

  6. Structural Characterisation of FabG from Yersinia pestis, a Key Component of Bacterial Fatty Acid Synthesis.

    Science.gov (United States)

    Nanson, Jeffrey D; Forwood, Jade K

    2015-01-01

    Ketoacyl-acyl carrier protein reductases (FabG) are ubiquitously expressed enzymes that catalyse the reduction of acyl carrier protein (ACP) linked thioesters within the bacterial type II fatty acid synthesis (FASII) pathway. The products of these enzymes, saturated and unsaturated fatty acids, are essential components of the bacterial cell envelope. The FASII reductase enoyl-ACP reductase (FabI) has been the focus of numerous drug discovery efforts, some of which have led to clinical trials, yet few studies have focused on FabG. Like FabI, FabG appears to be essential for survival in many bacteria, similarly indicating the potential of this enzyme as a drug target. FabG enzymes are members of the short-chain alcohol dehydrogenase/reductase (SDR) family, and like other SDRs, exhibit highly conserved secondary and tertiary structures, and contain a number of conserved sequence motifs. Here we describe the crystal structures of FabG from Yersinia pestis (YpFabG), the causative agent of bubonic, pneumonic, and septicaemic plague, and three human pandemics. Y. pestis remains endemic in many parts of North America, South America, Southeast Asia, and Africa, and a threat to human health. YpFabG shares a high degree of structural similarity with bacterial homologues, and the ketoreductase domain of the mammalian fatty acid synthase from both Homo sapiens and Sus scrofa. Structural characterisation of YpFabG, and comparison with other bacterial FabGs and the mammalian fatty acid synthase, provides a strong platform for virtual screening of potential inhibitors, rational drug design, and the development of new antimicrobial agents to combat Y. pestis infections.

  7. The Ubx Polycomb response element bypasses an unpaired Fab-8 insulator via cis transvection in Drosophila.

    Science.gov (United States)

    Lu, Danfeng; Li, Zhuoran; Li, Lingling; Yang, Liping; Chen, Guijun; Yang, Deying; Zhang, Yue; Singh, Vikrant; Smith, Sheryl; Xiao, Yu; Wang, Erlin; Ye, Yunshuang; Zhang, Wei; Zhou, Lei; Rong, Yikang; Zhou, Jumin

    2018-01-01

    Chromatin insulators or boundary elements protect genes from regulatory activities from neighboring genes or chromatin domains. In the Drosophila Abdominal-B (Abd-B) locus, the deletion of such elements, such as Frontabdominal-7 (Fab-7) or Fab-8 led to dominant gain of function phenotypes, presumably due to the loss of chromatin barriers. Homologous chromosomes are paired in Drosophila, creating a number of pairing dependent phenomena including transvection, and whether transvection may affect the function of Polycomb response elements (PREs) and thus contribute to the phenotypes are not known. Here, we studied the chromatin barrier activity of Fab-8 and how it is affected by the zygosity of the transgene, and found that Fab-8 is able to block the silencing effect of the Ubx PRE on the DsRed reporter gene in a CTCF binding sites dependent manner. However, the blocking also depends on the zygosity of the transgene in that the barrier activity is present when the transgene is homozygous, but absent when the transgene is heterozygous. To analyze this effect, we performed chromatin immunoprecipitation and quantitative PCR (ChIP-qPCR) experiments on homozygous transgenic embryos, and found that H3K27me3 and H3K9me3 marks are restricted by Fab-8, but they spread beyond Fab-8 into the DsRed gene when the two CTCF binding sites within Fab-8 were mutated. Consistent with this, the mutation reduced H3K4me3 and RNA Pol II binding to the DsRed gene, and consequently, DsRed expression. Importantly, in heterozygous embryos, Fab-8 is unable to prevent the spread of H3K27me3 and H3K9me3 marks from crossing Fab-8 into DsRed, suggesting an insulator bypass. These results suggest that in the Abd-B locus, deletion of the insulator in one copy of the chromosome could lead to the loss of insulator activity on the homologous chromosome, and in other loci where chromosomal deletion created hemizygous regions of the genome, the chromatin barrier could be compromised. This study highlights

  8. Parasitism and olfactory responses of Dastarcus helophoroides (Coleoptera: Bothrideridae) to different Cerambycid hosts

    Science.gov (United States)

    Jian-Rong Wei; Zhong-Qi Yang; Therese M. Poland; Jia-Wei. Du

    2009-01-01

    Dastarcus helophoroides (Fairmaire) (Coleoptera: Bothrideridae) is an important natural enemy of longhorned beetles (Coleoptera: Cerambycidae). It is distributed throughout most Provinces in China. We investigated whether there were differences among D. helophoroides populations collected from different hosts in different...

  9. /sup 99m/Tc labeling of antibodies to cardiac myosin Fab and to human fibrinogen

    International Nuclear Information System (INIS)

    Khaw, B.A.; Strauss, H.W.; Carvalho, A.; Locke, E.; Gold, H.K.; Haber, E.

    1982-01-01

    We have developed a method of labeling biologically active labile macromolecules, such as human fibrinogen (HF) and anticardiac-myosin Fab (AM-Fab), with /sup 99m/Tc at neutral pH. This method uses dithionite reduction of pertechnetate and subsequent labeling, to test the method with acid-labile macromolecules. Complexes of diethylene triamine pentaacetic acid with macromolecules such as human fibrinogen (D-HF) and anticardiac-myosin Fab (D-AM-Fab) were labeled and utilized in in vitro and in vivo studies. In biodistribution studies, the /sup 99m/Tc D-HF had a two-component blood clearance (half-times 1 hr and 15 hr) and was 80--88% coagulable. The /sup 99m/Tc AM-Fab retained its immunoreactivity as tested by affinity chromatography; also during in vivo localization in experimental myocardial infarction. This labeling technique provides an easy and efficient approach to the /sup 99m/Tc labeling of other biologically active and acid-labile macromolecules

  10. Reorienting the Fab domains of trastuzumab results in potent HER2 activators.

    Directory of Open Access Journals (Sweden)

    Justin M Scheer

    Full Text Available The structure of the Fab region of antibodies is critical to their function. By introducing single cysteine substitutions into various positions of the heavy and light chains of the Fab region of trastuzumab, a potent antagonist of HER2, and using thiol chemistry to link the different Fabs together, we produced a variety of monospecific F(ab'(2-like molecules with activities spanning from activation to inhibition of breast tumor cell growth. These isomers (or bis-Fabs of trastuzumab, with varying relative spatial arrangements between the Fv-regions, were able to either promote or inhibit cell-signaling activities through the PI3K/AKT and MAPK pathways. A quantitative phosphorylation mapping of HER2 indicated that the agonistic isomers produced a distinct phosphorylation pattern associated with activation. This study suggests that antibody geometric isomers, found both in nature and during synthetic antibody development, can have profoundly different biological activities independent of their affinities for their target molecules.

  11. Technetium-99m labeling of antibodies to cardiac myosin Fab and to human fibrinogen

    International Nuclear Information System (INIS)

    Khaw, B.A.; Strauss, H.W.; Carvalho, A.; Locke, E.; Gold, H.K.; Haber, E.

    1982-01-01

    A method of labeling biologically active labile macromolecules, such as human fibrinogen (HF) and anticardiac-myosin Fab (AM-Fab), with Tc-99m at neutral pH was developed. This method uses dithionite reduction of pertechnetate and subsequent labeling to test the method with acid-labile macromolecules. Complexes of diethylene triamine pentaacetic acid with macromolecules such as human fibrinogen (D-HF) and anticardiac-myosin Fab (D-AM-Fab) were labeled and utilized in in vitro and in vivo studies. In biodistribution studies, the Tc-99m D-HF had a two-component blood clearance (half-times 1 hr and 15 hr) and was 80-88% coagulable. The Tc-99m AM-Fab retained its immunoreactivity as tested by affinity chromatography; also during in vivo localization in experimental myocardial infarction. This labeling technique provides an easy and efficient approach to the Tc-99m labeling of other biologically active and acid-labile macromolecules

  12. Cyclization strategies of meditopes: affinity and diffraction studies of meditope–Fab complexes

    International Nuclear Information System (INIS)

    Bzymek, Krzysztof P.; Ma, Yuelong; Avery, Kendra A.; Horne, David A.; Williams, John C.

    2016-01-01

    An overview of cyclization strategies of a Fab-binding peptide to maximize affinity. Recently, a unique binding site for a cyclic 12-residue peptide was discovered within a cavity formed by the light and heavy chains of the cetuximab Fab domain. In order to better understand the interactions that drive this unique complex, a number of variants including the residues within the meditope peptide and the antibody, as well as the cyclization region of the meditope peptide, were created. Here, multiple crystal structures of meditope peptides incorporating different cyclization strategies bound to the central cavity of the cetuximab Fab domain are presented. The affinity of each cyclic derivative for the Fab was determined by surface plasmon resonance and correlated to structural differences. Overall, it was observed that the disulfide bond used to cyclize the peptide favorably packs against a hydrophobic ‘pocket’ and that amidation and acetylation of the original disulfide meditope increased the overall affinity ∼2.3-fold. Conversely, replacing the terminal cysteines with serines and thus creating a linear peptide reduced the affinity over 50-fold, with much of this difference being reflected in a decrease in the on-rate. Other cyclization methods, including the formation of a lactam, reduced the affinity but not to the extent of the linear peptide. Collectively, the structural and kinetic data presented here indicate that small perturbations introduced by different cyclization strategies can significantly affect the affinity of the meditope–Fab complex

  13. Fast conversion of scFv to Fab antibodies using type IIs restriction enzymes.

    Science.gov (United States)

    Sanmark, Hanna; Huovinen, Tuomas; Matikka, Tero; Pettersson, Tiina; Lahti, Maria; Lamminmäki, Urpo

    2015-11-01

    Single chain variable fragment (scFv) antibody libraries are widely used for developing novel bioaffinity reagents, although Fab or IgG molecules are the preferred antibody formats in many final applications. Therefore, rapid conversion methods for combining multiple DNA fragments are needed to attach constant domains to the scFv derived variable domains. In this study we describe a fast and easy cloning method for the conversion of single framework scFv fragments to Fab fragments using type IIS restriction enzymes. All cloning steps excluding plating of the Fab transformants can be done in 96 well plates and the procedure can be completed in one working day. The concept was tested by converting 69 scFv clones into Fab format on 96 well plates, which resulted in 93% success rate. The method is particularly useful as a high-throughput tool for the conversion of the chosen scFv clones into Fab molecules in order to analyze them as early as possible, as the conversion can significantly affect the binding properties of the chosen clones. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Cyclization strategies of meditopes: affinity and diffraction studies of meditope–Fab complexes

    Energy Technology Data Exchange (ETDEWEB)

    Bzymek, Krzysztof P.; Ma, Yuelong; Avery, Kendra A.; Horne, David A.; Williams, John C., E-mail: jcwilliams@coh.org [Beckman Research Institute of City of Hope, 1710 Flower Street, Duarte, CA 91010 (United States)

    2016-05-23

    An overview of cyclization strategies of a Fab-binding peptide to maximize affinity. Recently, a unique binding site for a cyclic 12-residue peptide was discovered within a cavity formed by the light and heavy chains of the cetuximab Fab domain. In order to better understand the interactions that drive this unique complex, a number of variants including the residues within the meditope peptide and the antibody, as well as the cyclization region of the meditope peptide, were created. Here, multiple crystal structures of meditope peptides incorporating different cyclization strategies bound to the central cavity of the cetuximab Fab domain are presented. The affinity of each cyclic derivative for the Fab was determined by surface plasmon resonance and correlated to structural differences. Overall, it was observed that the disulfide bond used to cyclize the peptide favorably packs against a hydrophobic ‘pocket’ and that amidation and acetylation of the original disulfide meditope increased the overall affinity ∼2.3-fold. Conversely, replacing the terminal cysteines with serines and thus creating a linear peptide reduced the affinity over 50-fold, with much of this difference being reflected in a decrease in the on-rate. Other cyclization methods, including the formation of a lactam, reduced the affinity but not to the extent of the linear peptide. Collectively, the structural and kinetic data presented here indicate that small perturbations introduced by different cyclization strategies can significantly affect the affinity of the meditope–Fab complex.

  15. Optimizing Production of Antigens and Fabs in the Context of Generating Recombinant Antibodies to Human Proteins.

    Directory of Open Access Journals (Sweden)

    Nan Zhong

    Full Text Available We developed and optimized a high-throughput project workflow to generate renewable recombinant antibodies to human proteins involved in epigenetic signalling. Three different strategies to produce phage display compatible protein antigens in bacterial systems were compared, and we found that in vivo biotinylation through the use of an Avi tag was the most productive method. Phage display selections were performed on 265 in vivo biotinylated antigen domains. High-affinity Fabs (<20nM were obtained for 196. We constructed and optimized a new expression vector to produce in vivo biotinylated Fabs in E. coli. This increased average yields up to 10-fold, with an average yield of 4 mg/L. For 118 antigens, we identified Fabs that could immunoprecipitate their full-length endogenous targets from mammalian cell lysates. One Fab for each antigen was converted to a recombinant IgG and produced in mammalian cells, with an average yield of 15 mg/L. In summary, we have optimized each step of the pipeline to produce recombinant antibodies, significantly increasing both efficiency and yield, and also showed that these Fabs and IgGs can be generally useful for chromatin immunoprecipitation (ChIP protocols.

  16. Waveband specific transcriptional control of select genetic pathways in vertebrate skin (Xiphophorus maculatus).

    Science.gov (United States)

    Walter, Ronald B; Boswell, Mikki; Chang, Jordan; Boswell, William T; Lu, Yuan; Navarro, Kaela; Walter, Sean M; Walter, Dylan J; Salinas, Raquel; Savage, Markita

    2018-05-10

    Evolution occurred exclusively under the full spectrum of sunlight. Conscription of narrow regions of the solar spectrum by specific photoreceptors suggests a common strategy for regulation of genetic pathways. Fluorescent light (FL) does not possess the complexity of the solar spectrum and has only been in service for about 60 years. If vertebrates evolved specific genetic responses regulated by light wavelengths representing the entire solar spectrum, there may be genetic consequences to reducing the spectral complexity of light. We utilized RNA-Seq to assess changes in the transcriptional profiles of Xiphophorus maculatus skin after exposure to FL ("cool white"), or narrow wavelength regions of light between 350 and 600 nm (i.e., 50 nm or 10 nm regions, herein termed "wavebands"). Exposure to each 50 nm waveband identified sets of genes representing discrete pathways that showed waveband specific transcriptional modulation. For example, 350-400 or 450-500 nm waveband exposures resulted in opposite regulation of gene sets marking necrosis and apoptosis (i.e., 350-400 nm; necrosis suppression, apoptosis activation, while 450-500 nm; apoptosis suppression, necrosis activation). Further investigation of specific transcriptional modulation employing successive 10 nm waveband exposures between 500 and 550 nm showed; (a) greater numbers of genes may be transcriptionally modulated after 10 nm exposures, than observed for 50 nm or FL exposures, (b) the 10 nm wavebands induced gene sets showing greater functional specificity than 50 nm or FL exposures, and (c) the genetic effects of FL are primarily due to 30 nm between 500 and 530 nm. Interestingly, many genetic pathways exhibited completely opposite transcriptional effects after different waveband exposures. For example, the epidermal growth factor (EGF) pathway exhibits transcriptional suppression after FL exposure, becomes highly active after 450-500 nm waveband exposure, and again, exhibits strong

  17. Targeting human prostate cancer with In-111-labeled D2B IgG, F(ab ')(2) and Fab fragments in nude mice with PSMA-expressing xenografts

    NARCIS (Netherlands)

    Lutje, Susanne; van Rij, Catharina M.; Franssen, Gerben M.; Fracasso, Giulio; Helfrich, Wijnand; Eek, Annemarie; Oyen, Wim J.; Colombatti, Marco; Boerman, Otto C.

    2014-01-01

    D2B is a new monoclonal antibody directed against an extracellular domain of prostate-specific membrane antigen (PSMA), which is overexpressed in prostate cancer. The potential of D2B IgG, and F(ab)(2) and Fab fragments of this antibody for targeting prostate cancer was determined in mice bearing

  18. Targeting human prostate cancer with (111) In-labeled D2B IgG, F(ab')2 and Fab fragments in nude mice with PSMA-expressing xenografts

    NARCIS (Netherlands)

    Lutje, S.; Rij, C.M. van; Franssen, G.M.; Fracasso, G.; Helfrich, W.; Eek, A.; Oyen, W.J.G.; Colombatti, M.; Boerman, O.C.

    2015-01-01

    D2B is a new monoclonal antibody directed against an extracellular domain of prostate-specific membrane antigen (PSMA), which is overexpressed in prostate cancer. The potential of D2B IgG, and F(ab')2 and Fab fragments of this antibody for targeting prostate cancer was determined in mice bearing

  19. Role of fibronectin in collagen deposition: Fab' to the gelatin-binding domain of fibronectin inhibits both fibronectin and collagen organization in fibroblast extracellular matrix

    OpenAIRE

    1982-01-01

    We report the effect of Fab' (anti-60k) to a 60,000 mol wt gelatin binding domain of fibronectin (1981, J. Biol. Chem. 256:5583) on diploid fibroblast (IMR-90) extracellular fibronectin and collagen organization. Anti-60k Fab' did not inhibit IMR-90 attachment or proliferation in fibronectin-depleted medium. Fibroblasts cultured with preimmune Fab' deposited a dense extracellular network of fibronectin and collagen detectable by immunofluorescence, while anti-60k Fab' prevented extracellular ...

  20. Ostrea puelchana (D'Orbigny 1842: a new host of Tumidotheres maculatus (Say, 1818 in northern Patagonia, Argentina Ostrea puelchana D’Orbigny 1842 nuevo hospedador de Tumidotheres maculatus (Say, 1818 en el norte de Patagonia, Argentina

    Directory of Open Access Journals (Sweden)

    María del Socorro Doldan

    2012-03-01

    Full Text Available Tumidotheres maculatus has been reported as a commensal of Mytilus platensis and Atrina seminuda and suggested as a parasite of Aequipecten tehuelchus in San Matías Gulf (SMG. The native flat oyster, Ostrea puelchana, has been studied in SMG for decades, and no crab-oyster association has been reported. In autumn-spring 2009 and 2010, oysters were collected from four banks in SMG, and the occurrence of T. maculatus inside the pallial cavity of O. puelchana was recorded. The prevalence varied between sites, with 0-12.5% at El Sótano, 0-16.7% at Caleta de los Loros, and no crabs found in the samples from Las Grutas and Puerto Lobos. Infected oysters hosted a single crab. Oysters were infested by either one male (47.7% or one female (15.1%. A positive but low correlation was found for male crab size and oyster size. The presence of these crabs inside the oysters could be due to the overlap of the distribution areas of O. puelchana and other bivalve hosts. Our findings may also be a consequence of local fishing. Commercial extraction of traditional hosts may have caused crabs to look for new hosts.En el Golfo San Matías (GSM, Tumidotheres maculatus ha sido registrado como comensal de Mytilus platensis y Atrina seminuda, y se lo ha indicado como parásito de Aequipecten tehuelchus. La ostra plana nativa Ostrea puelchana ha sido objeto de estudio por décadas en el GSM y no existen precedentes sobre una asociación entre las ostras y el cangrejo pinotérido. Durante otoño-primavera de 2009 y de 2010 se recolectaron ostras de cuatro bancos del GSM, registrándose la ocurrencia de T. maculatus en la cavidad paleal de O. puelchana. La prevalencia varió entre los sitios: 0-12,5% en El Sótano, 0-16,7% en Caleta de los Loros; no se encontraron cangrejos en los muestreos en Las Grutas y en Puerto Lobos. Se encontró un único cangrejo por hospedador; las ostras estuvieron infestadas por cangrejos macho (47,7%, o hembras (15,1%. Se encontró una

  1. Localization of 131I-labeled p97-specific Fab fragments in human melanoma as a basis for radiotherapy

    International Nuclear Information System (INIS)

    Larson, S.M.; Carrasquillo, J.A.; Krohn, K.A.

    1983-01-01

    33 patients with advanced malignant melanoma were studied after intravenous administration of 131 I-labeled Fab fragments specific for p97, an oncofetal glycoprotein of human melanoma. In all, 47 gamma camera imaging studies were performed for the purpose of localization of metastatic deposits. In addition to tumor, 131 I-Fab uptake was also seen in liver and kidney. 20 of these studies included simultaneous administration of both an 131 I-labeled Fab specific for p97, and an 125 I-labeled Fab not specific for p97. Blood clearance of p97-specific Fab was significantly more rapid than for nonspecific Fab. Eight of these patients had biopsies of subcutaneous nodules at 48 and 72 h postinjection in order to assess whether localization of radioactivity was antigen specific. Antigen-specific localization was observed with average ratios of specific/nonspecific uptake of 3.7 (48 h) and 3.4 (72 h); uptake was strongly correlated with tumor p97 concentration (r . 0.81, P less than 0.01). Also, imaging studies of the bio-distribution of 131 I-labeled anti-p97 Fab in patients selected for high p97 tumor concentration showed avid tumor uptake and more prolonged retention of labeled Fab in tumor than in normal tissues. Based on these studies, we estimated that total 131 I doses of 500 mCi could be safely given to patients before dose-limiting toxicity would be observed

  2. Detection of irradiation-induced, membrane heat shock protein 70 (Hsp70) in mouse tumors using Hsp70 Fab fragment

    International Nuclear Information System (INIS)

    Stangl, Stefan; Themelis, George; Friedrich, Lars; Ntziachristos, Vasilis; Sarantopoulos, Athanasios; Molls, Michael; Skerra, Arne; Multhoff, Gabriele

    2011-01-01

    Background and purpose: The major stress-inducible heat shock protein 70 (Hsp70) is frequently overexpressed in highly aggressive tumors, and elevated intracellular Hsp70 levels mediate protection against apoptosis. Following therapeutic intervention, such as ionizing irradiation, translocation of cytosolic Hsp70 to the plasma membrane is selectively increased in tumor cells and therefore, membrane Hsp70 might serve as a therapy-inducible, tumor-specific target structure. Materials and methods: Based on the IgG1 mouse monoclonal antibody (mAb) cmHsp70.1, we produced the Hsp70-specific recombinant Fab fragment (Hsp70 Fab), as an imaging tool for the detection of membrane Hsp70 positive tumor cells in vitro and in vivo. Results: The binding characteristics of Hsp70 Fab towards mouse colon (CT26) and pancreatic (1048) carcinoma cells at 4 deg. C were comparable to that of cmHsp70.1 mAb, as determined by flow cytometry. Following a temperature shift to 37 deg. C, Hsp70 Fab rapidly translocates into subcellular vesicles of mouse tumor cells. Furthermore, in tumor-bearing mice Cy5.5-conjugated Hsp70 Fab, but not unrelated IN-1 control Fab fragment (IN-1 ctrl Fab), gradually accumulates in CT26 tumors between 12 and 55 h after i.v. injection. Conclusions: In summary, the Hsp70 Fab provides an innovative, low immunogenic tool for imaging of membrane Hsp70 positive tumors, in vivo.

  3. Enoyl-Acyl Carrier Protein Reductase I (FabI) Is Essential for the Intracellular Growth of Listeria monocytogenes

    Science.gov (United States)

    Ericson, Megan E.; Frank, Matthew W.

    2016-01-01

    Enoyl-acyl carrier protein reductase catalyzes the last step in each elongation cycle of type II bacterial fatty acid synthesis and is a key regulatory protein in bacterial fatty acid synthesis. Genes of the facultative intracellular pathogen Listeria monocytogenes encode two functional enoyl-acyl carrier protein isoforms based on their ability to complement the temperature-sensitive growth phenotype of Escherichia coli strain JP1111 [fabI(Ts)]. The FabI isoform was inactivated by the FabI selective inhibitor AFN-1252, but the FabK isoform was not affected by the drug, as expected. Inhibition of FabI by AFN-1252 decreased endogenous fatty acid synthesis by 80% and lowered the growth rate of L. monocytogenes in laboratory medium. Robust exogenous fatty acid incorporation was not detected in L. monocytogenes unless the pathway was partially inactivated by AFN-1252 treatment. However, supplementation with exogenous fatty acids did not restore normal growth in the presence of AFN-1252. FabI inactivation prevented the intracellular growth of L. monocytogenes, showing that neither FabK nor the incorporation of host cellular fatty acids was sufficient to support the intracellular growth of L. monocytogenes. Our results show that FabI is the primary enoyl-acyl carrier protein reductase of type II bacterial fatty acid synthesis and is essential for the intracellular growth of L. monocytogenes. PMID:27736774

  4. Conversion of a Mouse Fab into a Whole Humanized IgG Antibody for Detecting Botulinum Toxin

    National Research Council Canada - National Science Library

    Palys, Thomas J; Schmid, Kara E; Scherer, John M; Schoepp, Randal J

    2006-01-01

    .... Therefore we sought to convert a murine Fab into a whole humanized IgG. The variable regions from an anti-botulinum Fab were cloned into human IgG heavy and light chain vectors and produced in myeloma cells...

  5. Efeitos de extratos alcoólicos de plantas sobre o caruncho do feijão vigna (Callosobruchus maculatus Effect of alcoholic extract of plants on weevil of cowpea (Callosobruchus maculatus

    Directory of Open Access Journals (Sweden)

    Francisco de A. C. Almeida

    2005-12-01

    Full Text Available Através de três métodos, extratos vegetais foram aplicados, ao Callosobruchus maculatus na fase adulta, inoculados ou não em uma massa de sementes, e na fase imatura (ovo com o objetivo de se controlar esta praga do feijão armazenado. Utilizaram-se flores, folhas, frutos e caule secos de oito espécies vegetais na obtenção dos extratos, em percolador, com solvente álcool etílico (70%. O delineamento experimental utilizado foi o inteiramente ao acaso, com os tratamentos distribuídos em esquema fatorial, cujos fatores quantitativos foram revelados pela regressão na análise de variância. Mediante os resultados obtidos, concluiu-se que a mortalidade dos insetos está relacionada com o tipo de extrato, os métodos de aplicação e com a dosagem aplicada, sendo os extratos de Callopogonium caeruleum e Piper nigrum os mais eficientes no controle do caruncho de feijão.Vegetable extracts were applied, through three methods, to the Callosobruchus maculatus in the adult phase, inoculated or not in a mass of seeds, in the immature phase (egg with the objective of controlling this pest of the stored beans. Dry flowers, leaves, fruits and dry stems of eight vegetable species were used to obtain the extracts in an extractor, with ethyl alcohol (70%. A completely randomized statistical design was used with the treatments distributed in a factorial scheme, the quantitative factors were analysed by the regression in the variance analysis. From the results obtained, it was concluded that the mortality of the insects is related to the extract type, the application methods and the applied dose, being the extracts of Callopogonium caeruleum and Piper nigrum the most efficient in the control of the weevil of cowpea.

  6. Development of 90Y-DOTA-nimotuzumab Fab fragment for radioimmunotherapy

    International Nuclear Information System (INIS)

    Alonso Martinez, L.M.; Marylaine Perez-Malo Cruz; Rene Leyva Montana; Calzada Falcon, V.N.; Minely Zamora Barrabi; Alejandro Arbesu Valdivia; Ignacio Hernandez Gonzalez; Mariela Leon Perez

    2014-01-01

    Yttrium-90-( 90 Y) labeled monoclonal antibodies prepared with a chelating agent, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), have been used for radioimmunotherapy of cancer. In the present work, the Fab fragment of anti-EGFR monoclonal antibody nimotuzumab was prepared with high purity, integrity and biological activity. The Fab fragment with high specific recognition of EGFR in NCI-H125 human lung adenocarcinoma cells was derivatized with DOTA-NHS applying a simple procedure. DOTA-nimotuzumab Fab fragment was successfully radiolabeled with 90 Y with high radiochemical yield. The in vitro stability of labeled product was optimal over 24 h in buffered solution at 37 deg C. Biodistribution and pharmacokinetic studies correctly evaluated the in vivo non-tumor uptake, dosage regimen and excretion pathway in normal Wistar rats. (author)

  7. 3D printing in social education: Eki-Fab and student PBL

    Science.gov (United States)

    Makino, Masato; Saito, Azusa; Kodama, Mai; Takamatsu, Kyuuichiro; Tamate, Hideaki; Sakai, Kazuyuki; Wada, Masato; Khosla, Ajit; Kawakami, Masaru; Furukawa, Hidemitsu

    2017-04-01

    Additive manufacturing or 3D printer is one of the most innovative material processing methods. We are considering that human resources for 3D printing would be needed in the future. To educate the abilities of the digital fabrication, we have the public digital fabrication space "Eki-Fab" for junior and high school students and Project Based Learning (PBL) class for undergraduate students. Eki-Fab is held on every Saturday at the Yonezawa train station. In the "Eki-Fab", anybody can study the utilizing of 3D printer and modeling technics under the instruction of staff in Yamagata University. In the PBL class, we have the class every Thursday. The students get the techniques of the digital fabrication through the PBL.

  8. Structure of Rotavirus Outer-Layer Protein VP7 Bound with a Neutralizing Fab

    Energy Technology Data Exchange (ETDEWEB)

    Aoki, Scott T.; Settembre, Ethan C.; Trask, Shane D.; Greenberg, Harry B.; Harrison, Stephen C.; Dormitzer, Philip R.; (Stanford-MED); (CH-Boston)

    2009-06-17

    Rotavirus outer-layer protein VP7 is a principal target of protective antibodies. Removal of free calcium ions (Ca{sup 2+}) dissociates VP7 trimers into monomers, releasing VP7 from the virion, and initiates penetration-inducing conformational changes in the other outer-layer protein, VP4. We report the crystal structure at 3.4 angstrom resolution of VP7 bound with the Fab fragment of a neutralizing monoclonal antibody. The Fab binds across the outer surface of the intersubunit contact, which contains two Ca{sup 2+} sites. Mutations that escape neutralization by other antibodies suggest that the same region bears the epitopes of most neutralizing antibodies. The monovalent Fab is sufficient to neutralize infectivity. We propose that neutralizing antibodies against VP7 act by stabilizing the trimer, thereby inhibiting the uncoating trigger for VP4 rearrangement. A disulfide-linked trimer is a potential subunit immunogen.

  9. 20 CFR 30.312 - What will the FAB do if the claimant objects to the recommended decision but does not request a...

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false What will the FAB do if the claimant objects....312 What will the FAB do if the claimant objects to the recommended decision but does not request a... period of time allotted in § 30.310 but does not request a hearing, the FAB will consider any objections...

  10. 20 CFR 30.318 - Can the FAB consider objections to HHS's reconstruction of a radiation dose or to the guidelines...

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false Can the FAB consider objections to HHS's... OF 2000, AS AMENDED Adjudicatory Process Hearings and Final Decisions on Claims § 30.318 Can the FAB... to HHS's reconstruction of the radiation dose to which the employee was exposed, the FAB will...

  11. 20 CFR 30.317 - Can the FAB request a further response from the claimant or return a claim to the district office?

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false Can the FAB request a further response from....317 Can the FAB request a further response from the claimant or return a claim to the district office? At any time before the issuance of its final decision, the FAB may request that the claimant submit...

  12. Imaging Potential Evaluation of Fab Derived from the Anti-EGFRvIII Monoclonal Antibody 4G1.

    Science.gov (United States)

    Jing, Shen; He, Yujia; He, Yanqiong; Wang, Liang; Jia, Jianhua; Shan, Xiaomin; Liu, Shuang; Tang, Min; Peng, Zhiping; Liu, Xujie

    2018-05-31

    As one of the most crucial epidermal growth factor receptor (EGFR) variants, EGFRvIII can be detected in various tumors but rarely in normal tissues, making it an ideal target for prognosis, diagnosis or immune therapy. The recently developed anti-EGFRvIII monoclonal antibody (mAb), 4G1, has been validated as a promising molecular probe to detect EGFRvIII expression in tumors by single-photon emission computed tomography/computed tomography imaging. To overcome shortcomings associated with the whole antibody, including long-term retention, circulation and enhanced permeability and retention effects, the Fab fragment of 4G1 (Fab-4G1) was generated, labeled with 131 I and evaluated in vitro and in vivo to test its potential application in molecular imaging. Whole mAb 4G1 was first digested by immobilized ficin and then purified through a protein A column to generate the Fab fragment, Fab-4G1. Next, SDS-PAGE, Western blot, indirect fluorescence assay, flow cytometry and enzyme-linked immunosorbent assay were performed to verify molecular weight, specificity and affinity of Fab-4G1. Finally, biodistribution planar gamma imaging was performed by injection of 131 I-labeled Fab-4G1 into xenografted EGFRvIII-overexpressed tumors in nude mice. Parallel studies were also performed with intact 4G1. The molecular weight of Fab was determined to be 35-40 kDa by SDS-PAGE. In vitro tests confirmed both intact 4G1 and Fab-4G1 specifically bound EGFRvIII but not wild-type EGFR, and Fab-4G1 showed decreased affinity. Compared to 131 I-4G1, biodistribution studies showed lower tumor uptake of 131 I-Fab-4G1 at all time points, but much faster elimination in all normal organs. As for planar gamma imaging, 131 I-Fab-4G1 and 31 I-4G1 showed similar imaging effect at 2 h after injection of tracer, while 131 I-Fab-4G1 was eliminated more quickly with time, suggesting radiolabeled Fab-4G1 could be potentially used for imaging of EGFRvIII-positive tumors at early time points. Radiolabeled

  13. High contrast tumor imaging with radio-labeled antibody Fab fragments tailored for optimized pharmacokinetics via PASylation.

    Science.gov (United States)

    Mendler, Claudia T; Friedrich, Lars; Laitinen, Iina; Schlapschy, Martin; Schwaiger, Markus; Wester, Hans-Jürgen; Skerra, Arne

    2015-01-01

    Although antigen-binding fragments (Fabs) of antibodies constitute established tracers for in vivo radiodiagnostics, their functionality is hampered by a very short circulation half-life. PASylation, the genetic fusion with a long, conformationally disordered amino acid chain comprising Pro, Ala and Ser, provides a convenient way to expand protein size and, consequently, retard renal filtration. Humanized αHER2 and αCD20 Fabs were systematically fused with 100 to 600 PAS residues and produced in E. coli. Cytofluorimetric titration analysis on tumor cell lines confirmed that antigen-binding activities of the parental antibodies were retained. The radio-iodinated PASylated Fabs were studied by positron emission tomography (PET) imaging and biodistribution analysis in mouse tumor xenograft models. While the unmodified αHER2 and αCD20 Fabs showed weak tumor uptake (0.8% and 0.2% ID/g, respectively; 24 h p.i.) tumor-associated radioactivity was boosted with increasing PAS length (up to 9 and 26-fold, respectively), approaching an optimum for Fab-PAS400. Remarkably, 6- and 5-fold higher tumor-to-blood ratios compared with the unmodified Fabs were measured in the biodistribution analysis (48 h p.i.) for αHER2 Fab-PAS100 and Fab-PAS200, respectively. These findings were confirmed by PET studies, showing high imaging contrast in line with tumor-to-blood ratios of 12.2 and 5.7 (24 h p.i.) for αHER2 Fab-PAS100 and Fab-PAS200. Even stronger tumor signals were obtained with the corresponding αCD20 Fabs, both in PET imaging and biodistribution analysis, with an uptake of 2.8% ID/g for Fab-PAS100 vs. 0.24% ID/g for the unmodified Fab. Hence, by engineering Fabs via PASylation, plasma half-life can be tailored to significantly improve tracer uptake and tumor contrast, thus optimally matching reagent/target interactions.

  14. Crotaline Fab antivenom appears to be effective in cases of severe North American pit viper envenomation: An integrative review

    Directory of Open Access Journals (Sweden)

    Mlynarchek Sara L

    2009-06-01

    Full Text Available Abstract Background In 2000, the United States Food and Drug Administration approved Crotalidae Polyvalent Immune Fab (Ovine (hereafter, FabAV, "for the management of patients with minimal to moderate North American Crotalid envenomation." Because whole-IgG pit viper antivenom is no longer available in the United States, FabAV is currently the only specific treatment option available to United States clinicians treating snakebite victims of any severity. No clinical trial data are available concerning the effectiveness of FabAV for treatment of severe snakebite, but several published articles describe its use in this setting. Methods We performed a comprehensive review of the English-language medical literature to identify all publications (1996 to July, 2008 containing data about the administration of FabAV. Two trained reviewers separately extracted case-level data concerning the administration of FabAV to patients with severe envenomation by North American crotaline snakes to a standardized form. Descriptive statistics were used. In addition, we hand-searched the US National Poison Data System reports for the years 2000–2006 to identify and describe any reports of death that occurred after FabAV administration. Results The literature review found 147 unique publications regarding FabAV. Twenty-four evaluable cases of severe human envenomation treated with FabAV were identified in 19 publications. Seven cases were described in five cohort studies, and 17 cases were described in 14 single patient case reports or non-cohort case series. Sixty-five specific severe venom effects were reported in these 24 patients, of which 50 effects (77% improved or resolved after FabAV therapy. Initial control of all severe venom effects was achieved in 12 patients (50%. The rate at which initial control was achieved was significantly higher among patients reported in the cohort series than in the case series and non-cohort reports (100% vs. 29%, P = 0

  15. EFEKTIVITAS VECTOBAC 12 AS (Bt H-14 DAN Bacillus thuringiensis H-14 TERHADAP VEKTOR MALARIA An. maculatus DI KOBAKAN DESA HARGOTIRTO, KECAMATAN KOKAP, KABUPATEN KULON PROGO

    Directory of Open Access Journals (Sweden)

    Blondine Ch. P.

    2012-09-01

    Full Text Available A study using Vectobac 12 AS ( Bt H-14 and Bacillus thuringiensis H-14 local strain was conducted at laboratory Vector and Reservoir Control Research Unit and breeding ponds of Anopheles maculatus in Kokap regency and Kulon Progo district. The objectives of this study are : (1. to detect the efficacy of B. thuringiensis H-14 local strain toward An. maculatus larvae at the laboratory. (2. to measure the effectiveness of B. thuringiensis H-14 local strain dosages 1 x LC95. 5 x LC95 and 10 x LC95 toward An. maculatus at the field. The efficacy test of B. thuringiensis H-14 local strain toward An. maculatus based on to the method proposed by WHO in order to determine the LC50 and LC90 which is computed using the probit analysis at the laboratory. The methods used R thuringiensis H-14 local strain dosages of 2.145 ppm (1 x LC95, 10.724 ppm (5 x LC95 and 21.448 ppm (10 x LC95 respectively were applied 8 ponds with the width of ponds ranging from 0.08 to 0.45 m2, 0.29 to 0.64 m2 and from 0.08-0.79 m2.The results showed, the dosages after 24 hours were 10.22 ppm (LC50, 27.11 ppm (LC90 and 35.75 ppm (LC95. After 48 hours the dosages were needed 7.74 ppm (LC50, 17.06 ppm (LC90 and 21.34 ppm (LC95, The effectiveness o/R thuringiensis H-14 local strain dosages of 2.145ppm (1 x LC95 toward An. maculatus larvae until 50 % survive the same time (7.35 days as B. thuringiensis H-14 (8.14 days dosages of 10.724 ppm (5 x LC95. B. thuringiensis H-14 local strain dosages of 21.448 ppm (10 x LC95 toward An. maculatus larvae until 50 % survive longer time (16.21 days than B. thuringiensis H-14 local strain 1 x LC95 and 5 x LC95 The B. thuringiensis H-14 local strain is effective for controlling mosquitoes larvae

  16. New synonymy in Cuban Tilloclytus (Coleoptera: Cerambycidae: Anaglyptini)

    Science.gov (United States)

    Examination of holotypes of Tilloclytus (Coleoptera: Cerambycidae: Anaglyptini) in the Fernando de Zayas collection (Havana, Cuba) and the Museum of Comparative Zoology, Harvard University reveals that T. elongatus Zayas (1975) is a new synonym of T. rufipes Fisher (1942)....

  17. QSAR models for prediction of chromatographic behavior of homologous Fab variants.

    Science.gov (United States)

    Robinson, Julie R; Karkov, Hanne S; Woo, James A; Krogh, Berit O; Cramer, Steven M

    2017-06-01

    While quantitative structure activity relationship (QSAR) models have been employed successfully for the prediction of small model protein chromatographic behavior, there have been few reports to date on the use of this methodology for larger, more complex proteins. Recently our group generated focused libraries of antibody Fab fragment variants with different combinations of surface hydrophobicities and electrostatic potentials, and demonstrated that the unique selectivities of multimodal resins can be exploited to separate these Fab variants. In this work, results from linear salt gradient experiments with these Fabs were employed to develop QSAR models for six chromatographic systems, including multimodal (Capto MMC, Nuvia cPrime, and two novel ligand prototypes), hydrophobic interaction chromatography (HIC; Capto Phenyl), and cation exchange (CEX; CM Sepharose FF) resins. The models utilized newly developed "local descriptors" to quantify changes around point mutations in the Fab libraries as well as novel cluster descriptors recently introduced by our group. Subsequent rounds of feature selection and linearized machine learning algorithms were used to generate robust, well-validated models with high training set correlations (R 2  > 0.70) that were well suited for predicting elution salt concentrations in the various systems. The developed models then were used to predict the retention of a deamidated Fab and isotype variants, with varying success. The results represent the first successful utilization of QSAR for the prediction of chromatographic behavior of complex proteins such as Fab fragments in multimodal chromatographic systems. The framework presented here can be employed to facilitate process development for the purification of biological products from product-related impurities by in silico screening of resin alternatives. Biotechnol. Bioeng. 2017;114: 1231-1240. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  18. Selective disulfide reduction for labeling and enhancement of Fab antibody fragments

    International Nuclear Information System (INIS)

    Kirley, Terence L.; Greis, Kenneth D.; Norman, Andrew B.

    2016-01-01

    Many methods have been developed for chemical labeling and enhancement of the properties of antibodies and their common fragments, including the Fab and F(ab’) 2 fragments. Somewhat selective reduction of some antibody disulfide bonds has been previously achieved, yielding antibodies and antibody fragments that can be labeled at defined sites, enhancing their utility and properties. Selective reduction of the two hinge disulfide bonds present in F(ab’) 2 fragments using mild reduction has been useful. However, such reduction is often not quantitative and results in the reduction of multiple disulfide bonds, and therefore subsequent multiple labeling or conjugation sites are neither homogenous nor stoichiometric. Here, a simple and efficient selective reduction of the single disulfide bond linking the partial heavy chain and the intact light chain which compose the Fab fragment is accomplished utilizing tris(2-carboxyethyl)phosphine (TCEP) immobilized on agarose beads. The resultant reduced cysteine residues were labeled with several cysteine-selective fluorescent reagents, as well as by cysteine-directed PEGylation. These two cysteine residues can also be re-ligated by means of a bifunctional cysteine cross-linking agent, dibromobimane, thereby both restoring a covalent linkage between the heavy and light chains at this site, far removed from the antigen binding site, and also introducing a fluorescent probe. There are many other research and clinical uses for these selectively partially reduced Fab fragments, including biotinylation, toxin and drug conjugation, and incorporation of radioisotopes, and this technique enables simple generation of very useful Fab fragment derivatives with many potential applications. - Highlights: • TCEP agarose is effective for selective reduction of a single Fab disulfide bond. • This disulfide is solvent accessible and distant from the antigen binding site. • A variety of buffers of varying pHs can be used, simplifying

  19. Selective disulfide reduction for labeling and enhancement of Fab antibody fragments.

    Science.gov (United States)

    Kirley, Terence L; Greis, Kenneth D; Norman, Andrew B

    2016-11-25

    Many methods have been developed for chemical labeling and enhancement of the properties of antibodies and their common fragments, including the Fab and F(ab') 2 fragments. Somewhat selective reduction of some antibody disulfide bonds has been previously achieved, yielding antibodies and antibody fragments that can be labeled at defined sites, enhancing their utility and properties. Selective reduction of the two hinge disulfide bonds present in F(ab') 2 fragments using mild reduction has been useful. However, such reduction is often not quantitative and results in the reduction of multiple disulfide bonds, and therefore subsequent multiple labeling or conjugation sites are neither homogenous nor stoichiometric. Here, a simple and efficient selective reduction of the single disulfide bond linking the partial heavy chain and the intact light chain which compose the Fab fragment is accomplished utilizing tris(2-carboxyethyl)phosphine (TCEP) immobilized on agarose beads. The resultant reduced cysteine residues were labeled with several cysteine-selective fluorescent reagents, as well as by cysteine-directed PEGylation. These two cysteine residues can also be re-ligated by means of a bifunctional cysteine cross-linking agent, dibromobimane, thereby both restoring a covalent linkage between the heavy and light chains at this site, far removed from the antigen binding site, and also introducing a fluorescent probe. There are many other research and clinical uses for these selectively partially reduced Fab fragments, including biotinylation, toxin and drug conjugation, and incorporation of radioisotopes, and this technique enables simple generation of very useful Fab fragment derivatives with many potential applications. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Analysis and functional consequences of increased Fab-sialylation of intravenous immunoglobulin (IVIG) after lectin fractionation.

    Science.gov (United States)

    Käsermann, Fabian; Boerema, David J; Rüegsegger, Monika; Hofmann, Andreas; Wymann, Sandra; Zuercher, Adrian W; Miescher, Sylvia

    2012-01-01

    It has been proposed that the anti-inflammatory effects of intravenous immunoglobulin (IVIG) might be due to the small fraction of Fc-sialylated IgG. In this study we biochemically and functionally characterized sialic acid-enriched IgG obtained by Sambucus nigra agglutinin (SNA) lectin fractionation. Two main IgG fractions isolated by elution with lactose (E1) or acidified lactose (E2) were analyzed for total IgG, F(ab')(2) and Fc-specific sialic acid content, their pattern of specific antibodies and anti-inflammatory potential in a human in vitro inflammation system based on LPS- or PHA-stimulated whole blood. HPLC and LC-MS testing revealed an increase of sialylated IgG in E1 and more substantially in the E2 fraction. Significantly, the increased amount of sialic acid residues was primarily found in the Fab region whereas only a minor increase was observed in the Fc region. This indicates preferential binding of the Fab sialic acid to SNA. ELISA analyses of a representative range of pathogen and auto-antigens indicated a skewed antibody pattern of the sialylated IVIG fractions. Finally, the E2 fraction exerted a more profound anti-inflammatory effect compared to E1 or IVIG, evidenced by reduced CD54 expression on monocytes and reduced secretion of MCP-1 (CCL2); again these effects were Fab- but not Fc-dependent. Our results show that SNA fractionation of IVIG yields a minor fraction (approx. 10%) of highly sialylated IgG, wherein the sialic acid is mainly found in the Fab region. The tested anti-inflammatory activity was associated with Fab not Fc sialylation.

  1. Analysis and functional consequences of increased Fab-sialylation of intravenous immunoglobulin (IVIG after lectin fractionation.

    Directory of Open Access Journals (Sweden)

    Fabian Käsermann

    Full Text Available It has been proposed that the anti-inflammatory effects of intravenous immunoglobulin (IVIG might be due to the small fraction of Fc-sialylated IgG. In this study we biochemically and functionally characterized sialic acid-enriched IgG obtained by Sambucus nigra agglutinin (SNA lectin fractionation. Two main IgG fractions isolated by elution with lactose (E1 or acidified lactose (E2 were analyzed for total IgG, F(ab'(2 and Fc-specific sialic acid content, their pattern of specific antibodies and anti-inflammatory potential in a human in vitro inflammation system based on LPS- or PHA-stimulated whole blood. HPLC and LC-MS testing revealed an increase of sialylated IgG in E1 and more substantially in the E2 fraction. Significantly, the increased amount of sialic acid residues was primarily found in the Fab region whereas only a minor increase was observed in the Fc region. This indicates preferential binding of the Fab sialic acid to SNA. ELISA analyses of a representative range of pathogen and auto-antigens indicated a skewed antibody pattern of the sialylated IVIG fractions. Finally, the E2 fraction exerted a more profound anti-inflammatory effect compared to E1 or IVIG, evidenced by reduced CD54 expression on monocytes and reduced secretion of MCP-1 (CCL2; again these effects were Fab- but not Fc-dependent. Our results show that SNA fractionation of IVIG yields a minor fraction (approx. 10% of highly sialylated IgG, wherein the sialic acid is mainly found in the Fab region. The tested anti-inflammatory activity was associated with Fab not Fc sialylation.

  2. Reference metrology in a research fab: the NIST clean calibrations thrust

    Science.gov (United States)

    Dixson, Ronald; Fu, Joe; Orji, Ndubuisi; Renegar, Thomas; Zheng, Alan; Vorburger, Theodore; Hilton, Al; Cangemi, Marc; Chen, Lei; Hernandez, Mike; Hajdaj, Russell; Bishop, Michael; Cordes, Aaron

    2009-03-01

    In 2004, the National Institute of Standards and Technology (NIST) commissioned the Advanced Measurement Laboratory (AML) - a state-of-the-art, five-wing laboratory complex for leading edge NIST research. The NIST NanoFab - a 1765 m2 (19,000 ft2) clean room with 743 m2 (8000 ft2) of class 100 space - is the anchor of this facility and an integral component of the new Center for Nanoscale Science and Technology (CNST) at NIST. Although the CNST/NanoFab is a nanotechnology research facility with a different strategic focus than a current high volume semiconductor fab, metrology tools still play an important role in the nanofabrication research conducted here. Some of the metrology tools available to users of the NanoFab include stylus profiling, scanning electron microscopy (SEM), and atomic force microscopy (AFM). Since 2001, NIST has collaborated with SEMATECH to implement a reference measurement system (RMS) using critical dimension atomic force microscopy (CD-AFM). NIST brought metrology expertise to the table and SEMATECH provided access to leading edge metrology tools in their clean room facility in Austin. Now, in the newly launched "clean calibrations" thrust at NIST, we are implementing the reference metrology paradigm on several tools in the CNST/NanoFab. Initially, we have focused on calibration, monitoring, and uncertainty analysis for a three-tool set consisting of a stylus profiler, an SEM, and an AFM. Our larger goal is the development of new and supplemental calibrations and standards that will benefit from the Class 100 environment available in the NanoFab and offering our customers calibration options that do not require exposing their samples to less clean environments. Toward this end, we have completed a preliminary evaluation of the performance of these instruments. The results of these evaluations suggest that the achievable uncertainties are generally consistent with our measurement goals.

  3. Improving methyl ketone production in Escherichia coli by heterologous expression of NADH-dependent FabG

    DEFF Research Database (Denmark)

    Goh, Ee Been; Chen, Yan; Petzold, Christopher J.

    2018-01-01

    balance, as fatty acid-derived pathways face the systematic metabolic challenge of net NADPH consumption (in large part, resulting from the key fatty acid biosynthetic enzyme FabG [β-ketoacyl-ACP reductase]) and net NADH production. In this study, we attempted to mitigate cofactor imbalance...... by heterologously expressing NADH-dependent, rather than NADPH-dependent, versions of FabG identified in previous studies. Of the four NADH-dependent versions of FabG tested in our previously best-reported methyl ketone-producing strain (EGS1895), the version from Acholeplasma laidlawii (Al_FabG) showed...... for the base strain (EGS1895) under fermentation conditions optimized in this study. Shotgun proteomic data for strains EGS2920 and EGS1895 during fed-batch fermentation were consistent with the goal of alleviating NADPH limitation through expression of Al_FabG. For example, relative to strain EGS1895, strain...

  4. Comparison of childhood myelodysplastic syndrome, AML FAB M6 or M7, CCG 2891: report from the Children's Oncology Group.

    Science.gov (United States)

    Barnard, Dorothy R; Alonzo, Todd A; Gerbing, Robert B; Lange, Beverly; Woods, William G

    2007-07-01

    Myelodysplastic syndromes (MDS), acute erythroleukemia (FAB M6), and acute megakaryocytic leukemia (FAB M7) have overlapping features. Children without Down syndrome or acute promyelocytic leukemia who were newly diagnosed with primary myelodysplastic syndrome or acute myeloid leukemia (AML) M6 or M7 were compared to children with de novo AML M0-M5. All children were entered on the Children's Cancer Group therapeutic research study CCG 2891. The presentation and outcomes of the 132 children diagnosed with MDS (60 children), AML FAB M6 (19 children), or AML FAB M7 (53 children) were similar. Children with AML FAB M7 were diagnosed at a significantly younger age (P = 0.001). Children with MDS, M6, or M7 had significantly lower white blood cell (WBC) counts (P = 0.001), lower peripheral blast counts (P M6 and AML M7 resemble MDS in presentation, poor induction success rates, and outcomes.

  5. Dietary shift in juvenile coral trout ( Plectropomus maculatus) following coral reef degradation from a flood plume disturbance

    Science.gov (United States)

    Wen, Colin K. C.; Bonin, Mary C.; Harrison, Hugo B.; Williamson, David H.; Jones, Geoffrey P.

    2016-06-01

    Acute environmental disturbances impact on habitat quality and resource availability, which can reverberate through trophic levels and become apparent in species' dietary composition. In this study, we observed a distinct dietary shift of newly settled and juvenile coral trout ( Plectropomus maculatus) following severe coral reef habitat degradation after a river flood plume affected the Keppel Islands, Australia. Hard coral cover declined by ~28 % in the 2 yr following the 2010-2011 floods, as did the abundance of young coral trout. Gut contents analysis revealed that diets had shifted from largely crustacean-based to non-preferred prey fishes following the disturbances. These results suggest that newly settled and juvenile coral trout modify their diet and foraging strategy in response to coral habitat degradation. This bottom-up effect of habitat degradation on the diet of a top coral reef predator may incur a metabolic cost, with subsequent effects on growth and survival.

  6. Influence of gamma radiation on oviposition and egg viability of Callosobruchus maculatus F. and grain loss in mungbean storage

    International Nuclear Information System (INIS)

    Dongre, T.K.; Rananavare, H.D.; Padwal Desai, S.R.

    1997-01-01

    Influence of gamma radiation on reproductive potential of Callosobruchus maculatus F. was studied by exposing them to 10, 30 and 50 Gy of radiation. Radiation at pupal stage was more effective in arresting egg laying and its further development. The dose of 50 Gy induced complete infertility in males when they were irradiated at pupal stage and only partial infertility was induced when irradiated at adult stage. In case of females whether irradiated at pupal or adult stage the dose of 50 Gy was sufficient to induce complete infertility. In order to assess the requirement of radiation doses for preventing the insect multiplication and subsequent damage in storage, mungbean samples with various developmental stages of insect were exposed to radiation at 50, 100 and 150 Gy. After three months of storage no grain damage was noticed when grains infested with different insect stages were irradiated with the dose of 100 or 150 Gy. (author)

  7. Process monitoring in high volume semiconductor production environment with in-fab TXRF

    International Nuclear Information System (INIS)

    Ghatak-Roy, A.R.; Hossain, T.Z.

    2000-01-01

    After its introduction in the 80's, TXRF has become an important tool for surface contamination analysis. This is particularly true for the semiconductor industries, where monitoring trace level contamination in ultra clean environment is absolutely necessary for successful device production with reasonable yield. In FAB 25 of the Advanced Micro Devices in Austin, we have installed two TXRF tools, which are model TXRF3750 manufactured by Rigaku. They contain rotating tungsten anodes with three beam capability for wide selection of elements. One of the beams (WM) is used for monitoring of low Z elements such as Na, Mg and Al. The standard output is 9 kW with 300 mA at 30 kV. The tool runs 24 hours a day, 7 days a week, except for maintenance and breakdowns. We have been using TXRF for in-fab monitoring of various tools and processes for trace contamination and some quantification of materials. This in-fab operation is important because it gives real time monitoring without the necessity of bringing the wafers out of the fab. Secondly, being in ultra clean fab environment, the risk of background contamination is minimized. Since TXRF measurement is fast and does not need any sample preparation, this works very well as production support tool. Several wafer fab technicians have been trained to use the tool for round the clock operation. We have successfully monitored tools and processes in our fab. One example is the monitoring of numerous sinks used in the cleaning of production wafers after various processes. Monitor wafers are run after sink cleaning and solvent changes and they are then analyzed for any contamination. Another example is the monitoring of tools that use Ferrofluidic seals so as to prevent any contamination from Fe and Cr. Other tools using TXRF include diffusion furnaces, etchers and plasma cleaning tools. We have also been monitoring processes such as ion implantation, metal deposition and rapid thermal annealing. In this presentation, we will

  8. Coleoptera species of forensic importance from Brazil: an updated list

    Directory of Open Access Journals (Sweden)

    Lúcia Massutti de Almeida

    2015-12-01

    Full Text Available ABSTRACT A list of the Coleoptera of importance from Brazil, based on published records was compiled. The checklist contains 345 species of 16 families allocated to 16 states of the country. In addition, three species of two families are registered for the first time. The fauna of Coleoptera of forensic importance is still not entirely known and future collection efforts and taxonomic reviews could increase the number of known species considerably in the near future.

  9. Immatures of Acanthocinini (Coleoptera, Cerambycidae, Lamiinae

    Directory of Open Access Journals (Sweden)

    Sônia A. Casari

    2014-06-01

    Full Text Available Immatures of Acanthocinini (Coleoptera, Cerambycidae, Lamiinae. Larva and pupa of Eutrypanus dorsalis (Germar, 1928, collected in trunks of Pinus elliottii Engelm., and Paratenthras martinsi Monné, 1998, collected in spathes of Scheelea phalerata (Mart. ex Spreng. Burret, are described and illustrated. Larva and pupa of Lophopoeum timbouvae Lameere, 1884, collected in Hymenaea corbaril L., Enterolobium contortisiliquum (Vell. Morong and Pterogyne nitens Tul., are redescribed and illustrated. A table with all described immatures of Lamiinae, and a comparison among the immatures of Acanthocinini are presented. Biological notes and new records are also included.

  10. Similar effectiveness of Fab and F(ab')2 antivenoms in the neutralization of hemorrhagic activity of Vipera berus snake venom in mice

    OpenAIRE

    Lomonte, Bruno; León Montero, Guillermo; Hanson, Lars Ake

    1996-01-01

    The ability of two antivenoms to Vipera spp., consisting of Fab (Therapeutic Antibodies), or of F(ab′)2 (Zagreb Institute of Immunology) antibody fragments, to neutralize the hemorrhagic activity of Vipera berus snake venom in mice, was compared. First, the neutralizing potency was determined by in vitro preincubation of venom and antivenom, followed by intradermal injection into mice and subsequent measurement of the hemorrhagic area. Both antivenoms had the same anti-hemorrhagic potency, in...

  11. AGILE integration into APC for high mix logic fab

    Science.gov (United States)

    Gatefait, M.; Lam, A.; Le Gratiet, B.; Mikolajczak, M.; Morin, V.; Chojnowski, N.; Kocsis, Z.; Smith, I.; Decaunes, J.; Ostrovsky, A.; Monget, C.

    2015-09-01

    mix logic Fab) in term of product and technology portfolio AGILE corrects for up to 120nm of product topography error on process layer with less than 50nm depth of focus Based on tool functionalities delivered by ASML and on high volume manufacturing requirement, AGILE integration is a real challenge. Regarding ST requirements "Automatic AGILE" functionality developed by ASML was not a turnkey solution and a dedicated functionality was needed. A "ST homemade AGILE integration" has been fully developed and implemented within ASML and ST constraints. This paper describes this integration in our Advanced Process Control platform (APC).

  12. High quality mask storage in an advanced Logic-Fab

    Science.gov (United States)

    Jähnert, Carmen; Fritsche, Silvio

    2012-02-01

    High efficient mask logistics as well as safe and high quality mask storage are essential requirements within an advanced lithography area of a modern logic waferfab. Fast operational availability of the required masks at the exposure tool with excellent mask condition requires a safe mask handling, safeguarding of high mask quality over the whole mask usage time without any quality degradation and an intelligent mask logistics. One big challenge is the prevention of haze on high advanced phase shift masks used in a high volume production line for some thousands of 248nm or 193nm exposures. In 2008 Infineon Dresden qualified a customer specific developed semi-bare mask storage system from DMSDynamic Micro Systems in combination with a high advanced mask handling and an interconnected complex logistic system. This high-capacity mask storage system DMS M1900.22 for more than 3000 masks with fully automated mask and box handling as well as full-blown XCDA purge has been developed and adapted to the Infineon Lithotoollandscape using Nikon and SMIF reticle cases. Advanced features for ESD safety and mask security, mask tracking via RFID and interactions with the exposure tools were developed and implemented. The stocker is remote controlled by the iCADA-RSM system, ordering of the requested mask directly from the affected exposure tool allows fast access. This paper discusses the advantages and challenges for this approach as well as the practical experience gained during the implementation of the new system which improves the fab performance with respect to mask quality, security and throughput. Especially the realization of an extremely low and stable humidity level in addition with a well controlled air flow at each mask surface, preventing masks from haze degradation and particle contamination, turns out to be a notable technical achievement. The longterm stability of haze critical masks has been improved significantly. Relevant environmental parameters like

  13. New species of Cerambycinae from the Neotropical Region, and nomen novum for Anelaphus maculatus Galileo, Martins, and Santos-Silva, 2014 (Elaphidiini).

    Science.gov (United States)

    Galileo, Maria Helena M; Martins, Ubirajara R; Santos-Silva, Antonio

    2015-07-17

    Six new species and one new genus are described: Criodion spinosum sp. nov. (Cerambycini), from Bolivia; Eburodacrys wappesi sp. nov. and Eburodacrys skillmani sp. nov. (Eburiini), from Bolivia; Eupempelus rileyorum sp. nov. (Heteropsini) from Panama; Sphalloeme mexicana sp. nov. (Oemini), from Mexico; Wappesoeme camiri sp. nov., new genus (Oemini), from Bolivia. Wappesoeme, Eburodacrys wappesi, E. skillmani, Eupempelus rileyorum, and Criodion spinosum are included in previously published keys. Anelaphus erakyra nomen novum for A. maculatus Galileo et al., 2014 is established.

  14. In vitro Fab display: a cell-free system for IgG discovery

    Science.gov (United States)

    Stafford, Ryan L.; Matsumoto, Marissa L.; Yin, Gang; Cai, Qi; Fung, Juan Jose; Stephenson, Heather; Gill, Avinash; You, Monica; Lin, Shwu-Hwa; Wang, Willie D.; Masikat, Mary Rose; Li, Xiaofan; Penta, Kalyani; Steiner, Alex R.; Baliga, Ramesh; Murray, Christopher J.; Thanos, Christopher D.; Hallam, Trevor J.; Sato, Aaron K.

    2014-01-01

    Selection technologies such as ribosome display enable the rapid discovery of novel antibody fragments entirely in vitro. It has been assumed that the open nature of the cell-free reactions used in these technologies limits selections to single-chain protein fragments. We present a simple approach for the selection of multi-chain proteins, such as antibody Fab fragments, using ribosome display. Specifically, we show that a two-chain trastuzumab (Herceptin) Fab domain can be displayed in a format which tethers either the heavy or light chain to the ribosome while retaining functional antigen binding. Then, we constructed synthetic Fab HC and LC libraries and performed test selections against carcinoembryonic antigen (CEA) and vascular endothelial growth factor (VEGF). The Fab selection output was reformatted into full-length immunoglobulin Gs (IgGs) and directly expressed at high levels in an optimized cell-free system for immediate screening, purification and characterization. Several novel IgGs were identified using this cell-free platform that bind to purified CEA, CEA positive cells and VEGF. PMID:24586053

  15. Impact of Fab Lab Tulsa on Student Self-Efficacy toward STEM Education

    Science.gov (United States)

    Dubriwny, Nicholas; Pritchett, Nathan; Hardesty, Michelle; Hellman, Chan M.

    2016-01-01

    Student self-confidence is important to any attempt to increase interest and achievement in Science, Technology, Engineering, and Math (STEM) education. This study presents a longitudinal examination of Fab Lab Tulsa's impact on attitude and self-efficacy toward STEM education among middle-school aged students. Paired samples t-test showed a…

  16. Impact of surfactants on the target recognition of Fab-conjugated PLGA nanoparticles.

    Science.gov (United States)

    Kennedy, Patrick J; Perreira, Ines; Ferreira, Daniel; Nestor, Marika; Oliveira, Carla; Granja, Pedro L; Sarmento, Bruno

    2018-06-01

    Targeted drug delivery with nanoparticles (NPs) requires proper surface ligand presentation and availability. Surfactants are often used as stabilizers in the production of targeted NPs. Here, we evaluated the impact of surfactants on ligand functionalization and downstream molecular recognition. Our model system consisted of fluorescent poly(lactic-co-glycolic acid) (PLGA) NPs that were nanoprecipitated in one of a small panel of commonly-used surfactants followed by equivalent washes and conjugation of an engineered Fab antibody fragment. Size, polydispersity index and zeta potential were determined by dynamic light scattering and laser Doppler anemometry, and Fab presence on the NPs was assessed by enzyme-linked immunosorbent assay. Most importantly, Fab-decorated NP binding to the cell surface receptor was monitored by fluorescence-activated cell sorting. 2% polyvinyl alcohol, 1% sodium cholate, 0.5% Pluronic F127 (F127) and 2% Tween-80 were initially tested. Of the four surfactants tested, PLGA NPs in 0.5% F127 and 2% Tween-80 had the highest cell binding. These two surfactants were then retested in two different concentrations, 0.5% and 2%. The Fab-decorated PLGA NPs in 2% F127 had the highest cell binding. This study highlights the impact of common surfactants and their concentrations on the downstream targeting of ligand-decorated NPs. Similar principles should be applied in the development of future targeted nanosystems where surfactants are employed. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. Wafer-level packaged RF-MEMS switches fabricated in a CMOS fab

    NARCIS (Netherlands)

    Tilmans, H.A.C.; Ziad, H.; Jansen, Henricus V.; Di Monaco, O.; Jourdain, A.; De Raedt, W.; Rottenberg, X.; De Backer, E.; Decoussernaeker, A.; Baert, K.

    2001-01-01

    Reports on wafer-level packaged RF-MEMS switches fabricated in a commercial CMOS fab. Switch fabrication is based on a metal surface micromachining process. A novel wafer-level packaging scheme is developed, whereby the switches are housed in on-chip sealed cavities using benzocyclobutene (BCB) as

  18. The Emerging Importance of IgG Fab Glycosylation in Immunity

    NARCIS (Netherlands)

    van de Bovenkamp, Fleur S.; Hafkenscheid, Lise; Rispens, Theo; Rombouts, Yoann

    2016-01-01

    Human IgG is the most abundant glycoprotein in serum and is crucial for protective immunity. In addition to conserved IgG Fc glycans, ∼15-25% of serum IgG contains glycans within the variable domains. These so-called "Fab glycans" are primarily highly processed complex-type biantennary N-glycans

  19. Fab glycosylation of immunoglobulin G does not associate with improvement of rheumatoid arthritis during pregnancy

    NARCIS (Netherlands)

    A. Bondt (Albert); M. Wuhrer (Manfred); T.M. Kuijper (Martijn); J.M.W. Hazes (Mieke); R.J.E.M. Dolhain (Radboud)

    2016-01-01

    textabstractBackground: Changes in immunoglobulin G (IgG) constant domain (Fc) glycosylation are associated with changes in rheumatoid arthritis (RA) disease activity in response to pregnancy. Here, we sought to determine whether the same holds true for variable domain (Fab) glycosylation. Methods:

  20. Reorienting the Fab Domains of Trastuzumab Results in Potent HER2 Activators

    Science.gov (United States)

    Scheer, Justin M.; Sandoval, Wendy; Elliott, J. Michael; Shao, Lily; Luis, Elizabeth; Lewin-Koh, Sock-Cheng; Schaefer, Gabriele; Vandlen, Richard

    2012-01-01

    The structure of the Fab region of antibodies is critical to their function. By introducing single cysteine substitutions into various positions of the heavy and light chains of the Fab region of trastuzumab, a potent antagonist of HER2, and using thiol chemistry to link the different Fabs together, we produced a variety of monospecific F(ab′)2-like molecules with activities spanning from activation to inhibition of breast tumor cell growth. These isomers (or bis-Fabs) of trastuzumab, with varying relative spatial arrangements between the Fv-regions, were able to either promote or inhibit cell-signaling activities through the PI3K/AKT and MAPK pathways. A quantitative phosphorylation mapping of HER2 indicated that the agonistic isomers produced a distinct phosphorylation pattern associated with activation. This study suggests that antibody geometric isomers, found both in nature and during synthetic antibody development, can have profoundly different biological activities independent of their affinities for their target molecules. PMID:23284778

  1. Advance in Research on Mycobacterium tuberculosis FabG4 and Its Inhibitor

    Directory of Open Access Journals (Sweden)

    Debajyoti Dutta

    2018-06-01

    Full Text Available Increasing evidence from recent reports of drug-resistant mycobacterial strains poses a challenge worldwide. Drug-resistant strains often undergo mutations, adopt alternative pathways, and express drug efflux pumps to reduce or eliminate drug doses. Besides these intrinsic resistance mechanisms, bacteria can evade drug doses by forming biofilms. Biofilms are the concerted growth of adherent microorganisms, which can also be formed at the air-water interface. The growth is supported by the extracellular polymer matrix which is self-produced by the microorganisms. Reduced metabolic activity in a nutrient-deficient environment in the biofilm may cause the microorganisms to take alternative pathways that can make the microorganisms recalcitrant to the drug doses. Recent works have shown that Mycobacterium tuberculosis expresses several proteins during its growth in biofilm, those when deleted, did not show any effect on mycobacterial growth in normal nutrient-sufficient conditions. Studying these unconventional proteins in mycobacterial biofilms is therefore of utmost importance. In this article, I will discuss one such mycobacterial biofilm-related protein FabG4 that is recently shown to be important for mycobacterial survival in the presence of antibiotic stressors and limited nutrient condition. In an attempt to find more effective FabG4 inhibitors and its importance in biofilm forming M. tuberculosis, present knowledge about FabG4 and its known inhibitors are discussed. Based on the existing data, a putative role of FabG4 is also suggested.

  2. The preparation of F(ab')2 fragment and it's application in tumor radioimmunoimaging

    International Nuclear Information System (INIS)

    Yang Ziyi

    1991-01-01

    Monoclonal antibody against lung cancer was digested into F(ab') 2 fragment by pepsin and papain digestion. The yields of pure F(ab') 2 were 32.3 ± 5.5% and 52.3 ± 12.0% respectively. The immunoreactivity of F(ab') 2 was based on the ELISA assay and the cell binding studies was retained well. In the localization experiments, radioiodinated F(ab') 2 was injected intraperitoneally into the nude mice bearing human xenografts of lung cancer. The highest radioactivity in tumors, 1.37% of injected dose per gram, was reached on the first day after injection; its T/NT ratios were higher than those of the intact IgG in all tissues except kidney. The localization index (LI) in tumors was 4.95, while the average LI value of normal tissues was 1.25. After the injection of 131 I-F(ab') 2 intraperitoneally into lung tumor-bearing nude mice, photo scintigraphy was performed at intervals of 12 hrs. The xenografts were visualized distinctly during 36 ∼ 48 hr, and the nonspecific background was very low at 48 hr

  3. Fab antibody fragment-functionalized liposomes for specific targeting of antigen-positive cells

    Czech Academy of Sciences Publication Activity Database

    Ohradanova-Repic, A.; Nogueira, E.; Hartl, I.; Gomes, A.C.; Preto, A.; Steinhuber, E.; Muehlgrabner, V.; Repic, M.; Kuttke, M.; Zwirzitz, A.; Prouza, M.; Suchánek, M.; Wozniak-Knopp, G.; Hořejší, Václav; Schabbauer, G.; Cavaco-Paulo, A.; Stockinger, H.

    2018-01-01

    Roč. 14, č. 1 (2018), s. 123-130 ISSN 1549-9634 Institutional support: RVO:68378050 Keywords : Active targeting * Liposome functionalization * Immunoliposome * Antibody engineering * Recombinant Fab antibody fragment Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Cell biology Impact factor: 5.720, year: 2016

  4. Development of radioinmunoconjugate 90Y-DOTA-nimotuzumab-Fab for therapy of EGFR over expressing tumors

    International Nuclear Information System (INIS)

    Rodriguez, A. M.; Alonso, L. M.; Gongora, M.; Leyva, R.; Solana, A.

    2015-01-01

    Many monoclonal antibodies conjugated with 1,4,7,10-tetraaza cyclododecane-N, N', N'', N'''-tetraacetic acid (DOTA) and radiolabeled with 90 Y, have been used for radioimmunotherapy. As know IgG molecules are heavy proteins with a molecular weight of approximately 150 kDa. Accordingly, intact IgG antibodies may have significant slow kinetics biodistribution and severely limited properties of tissue penetration. Antibody fragments labeled with radio metals could be promising radiopharmaceuticals for imaging and non-invasive therapy due to its high affinity to the tumor, the lack of effector function and rapid pharmacokinetic. In this work, the nimotuzumab Fab fragment was obtained by cleavage with papain in molar excess. After separating the reaction mixture in three steps using affinity, size exclusion and ion exchange chromatography; the Fab fragment showed high values of purity, integrity and identity. The Fab fragment was derivatized with DOTA and labeled with 90 Y. The radioimmunoconjugate with high radiochemical yield was assessed by in vitro stability with an excess of 50mM DTPA. The development of 90 Y-DOTA-Nimotuzumab-Fab radioimmunoconjugate allows to count on as a potential agent for radioimmunotherapy. (Author)

  5. The evolution of asymmetric genitalia in Coleoptera.

    Science.gov (United States)

    Schilthuizen, Menno; de Jong, Paulien; van Beek, Rick; Hoogenboom, Tamara; Schlochtern, Melanie Meijer Zu

    2016-12-19

    The evolution of asymmetry in male genitalia is a pervasive and recurrent phenomenon across almost the entire animal kingdom. Although in some taxa the asymmetry may be a response to the evolution of one-sided, male-above copulation from a more ancestral female-above condition, in other taxa, such as Mammalia and Coleoptera, this explanation appears insufficient. We carried out an informal assessment of genital asymmetry across the Coleoptera and found that male genital asymmetry is present in 43% of all beetle families, and at all within-family taxonomic levels. In the most diverse group, Cucujiformia, however, genital asymmetry is comparatively rare. We also reconstructed the phylogeny of the leiodid tribe Cholevini, and mapped aspects of genital asymmetry on the tree, revealing that endophallus sclerites, endophallus, median lobe and parameres are, in a nested fashion, increasingly unlikely to have evolved asymmetry. We interpret these results in the light of cryptic female choice versus sexually antagonistic coevolution and advocate further ways in which the phenomenon may be better understood.This article is part of the themed issue 'Provocative questions in left-right asymmetry'. © 2016 The Authors.

  6. Effect of some pulverised plant materials on the developmental stages of fish beetle, Dermestes maculatus Degeer in smoked catfish (Clarias gariepinus) during storage.

    Science.gov (United States)

    Fasakin, E A; Aberejo, B A

    2002-11-01

    The effectiveness of pulverised plant materials; Tithonium diversifolia, Afromomum melegueta, Nicotiana tabacum, Monodora myristica and Piper guineense as ovicidal, larvicidal and adult deterents of fish beetle (Dermestes maculatus) in smoked catfish (Clarias gariepinus) during storage were evaluated. Leaves of T. diversifolia, N. tabacum and seeds of A. melegueta, M. myristica and P. guineese were dried and pulverised into powder. Adults and larvae of third generation (F3) of D. maculatus were introduced into Kilner jars containing disinfested fish samples. Pulverised plant materials were applied to the surface of the fish samples at 10% (w/w) and monitored for 40 days, while egg hatchability of the insects was monitored for seven days. The result showed that all the plant materials had varying degree of insecticidal activities. Pulverised powder of P. guineense and A. melegueta were the most effective and significantly (P 0.05) affected by the plant materials. The percentage weight loss in fish treated with P. guineense and A. melegueta were minimal compared with the untreated fish sample. The result of this study showed that pulverised plant materials obtained from P. guineense and A. melegueta could be used to deter egg hatchability and adult emergence of D. maculatus in smoked catfish during storage. This could also reduce percentage losses due to insect infestation on smoked fish during storage.

  7. Evaluation of the efficacy of Levo 2.4 Sl against the cowpea beetle, Callosobruchus maculatus (F.) on stored cowpea grains

    International Nuclear Information System (INIS)

    Osekre, E. A.; Abrahamani, J.; Abudu, D.T.

    2013-01-01

    The adverse effects of synthetic pesticides on the environment and non-target organisms have necessitated the exploration of other safer and equally effective pesticides for the management of pests. The efficacy of Levo 2.4 SL (Oxymatrine 2.4 SL; Prosular oxamatrine a. i.), a botanical, against Callosobruchus maculatus (F.) infestation on stored cowpea, Vigna unguiculata (L.) (Walp.) was investigated in the insectary of the Entomology Section of the Faculty of Agriculture of the Kwame Nkrumah University of Science and Technology. The effects of Levo on the adult mortality, ovipoistion and F1 progeny of the insect were evaluated. Thus 200 g of cowpea grains were treated with three dosages; 0.01 ml, 0.02 ml and 0.03 ml of Levo 2.4 SL. Betallic Supper 2.5 EC (Primophos methyl and Permethrin a. i.) was used as a check and a control (untreated grains) was also maintained. The mean number of eggs laid and the number of F1 adults that emerged were significantly (P < 0.05) smaller in the Levo treated grains than the control. The higher dosages of Levo 2.4 SL were more effective against C. maculatus infestation and damage. At the end of four months of storage, Levo at all the dosages offered complete protection to the cowpea grains. Levo 2.4 SL at a dosage of 0.02 ml is considered the right dosage that can give optimum protection to cowpea grains against C. maculatus. (au)

  8. Preparation and radiolabeling of humanized anti-HER1 monoclonal antibody nimotuzumab Fab' fragment with 68Ga and 90Y

    International Nuclear Information System (INIS)

    Alonso Martinez, L. M.; Xiques Castillo, A.; Leyva Montanna, R.; Perez-Malo Cruz, M.; Zamora Barrabi, M.; Manresa Sanchez, Y.

    2013-01-01

    Antibody-based targeted delivery of radioisotopes to malignant tissues is a promising approach in cancer diagnostics and therapy. However, intact antibody molecules are large glycoproteins (∼150 kDa) that have limited application in molecular imaging and therapy due to their relatively slow clearance from the circulation leading to a high background signal rather both cases the sensitivity can be increased with the use of enzymatically produced Fab' fragments. In this work, the ability to get labeled with 62 Ga and 90 Y of a monoclonal antibody (mAb) Fab' fragment against the transmembrane receptor tyrosine kinase HER-1 was studied for future applications in PET imaging and radioimmunotherapy of tumors. In order to obtain the Fab' fragment the mAb was cleaved with pepsin in molar excess. After separating the reaction mixture in two steps using affinity and ion-exchange chromatography, the Fab' fragment was finally obtained by reduction of the F(ab') 2 with a molar excess of 2-mercaptoethanol followed by a size exclusion purification step. The Fab' fragment was derivatized with 1,4,7,10-tetraaza cyclododecane-1,4,7,10-tetraacetic acid mono N-hydroxysuccinimide commercial ester (DOTA-NHS-ester) applying a simple procedure and the number of DOTA groups linked to Fab' were determinate. The labeling of the conjugate with 68 Ga and 90 Y from 'in-house generators yielded radiochemically pure probes that can become a suitable radioimmunoconjugated in a near future. (Author)

  9. Application of 99mTc-labeled chimeric Fab fragments of monoclonal antibody A7 for radioimmunoscintigraphy of pancreatic cancer

    International Nuclear Information System (INIS)

    Matsumura, Hiroomi

    1999-01-01

    Pancreatic cancer is one of the most lethal diseases and its prognosis is still poor. To improve the survival rate, it is essential to develop new technologies for early and definitive diagnosis. In this study, chimeric Fab fragments of monoclonal antibody A7 were successfully radio-labeled with 99m Tc, preventing depression of the antigen-binding activity. 99m Tc-labeled monoclonal antibody A7, 99m Tc-labeled chimeric Fab fragments of monoclonal antibody A7, 99m Tc-labeled normal mouse IgG and 99m Tc-labeled Fab fragments of normal mouse IgG were injected intravenously into nude mice bearing human pancreatic cancer xenografts and the radioactivity was subsequently measured. The tumor accumulation was significantly higher with labeled monoclonal antibody A7 than with normal mouse IgG, and higher with chimeric Fab fragments of monoclonal antibody A7 than with Fab fragments of normal mouse IgG. The tumor/blood ratio of radioactivity increased rapidly over time with chimeric Fab fragments of monoclonal antibody A7. These results suggest that chimeric Fab fragments of monoclonal antibody A7 may be useful for diagnosing pancreatic cancer by means of radioimmunoscintigraphy. (author)

  10. Effectiveness of Alpha-toxin Fab Monoclonal Antibody Therapy in Limiting the Pathology of Staphylococcus aureus Keratitis.

    Science.gov (United States)

    Caballero, Armando R; Foletti, Davide L; Bierdeman, Michael A; Tang, Aihua; Arana, Angela M; Hasa-Moreno, Adela; Sangalang, Emma Ruth B; O'Callaghan, Richard J

    2015-08-01

    To investigate the effectiveness of a high-affinity human monoclonal antibody Fab fragment to Staphylococcus aureus alpha-toxin (LTM14 Fab) as therapy for S. aureus keratitis. A single topical drop of the LTM14 Fab antibody to alpha-toxin alone, or in 0.006% benzalkonium chloride (BAK), was applied every 30 min to S. aureus-infected rabbit corneas from 9 to 14 hours post-infection. Erosions and pathology were measured at 15 h post-infection. LTM14 Fab with BAK limited corneal erosions better than LTM14 Fab alone (p = 0.036), and both limited erosions compared to untreated eyes (p ≤ 0.0001). Overall pathology was similar in all groups (p ≥ 0.070), but iritis and chemosis were reduced by treatment (p ≤ 0.036). The high-affinity human monoclonal Fab fragment antibody (LTM14 Fab) to S. aureus alpha-toxin was effective in reducing corneal damage during S. aureus keratitis.

  11. X-ray studies of recombinant anti-testosterone Fab fragments: the use of PEG 3350 in crystallization.

    Science.gov (United States)

    Valjakka, J; Hemminki, A; Teerinen, T; Takkinen, K; Rouvinen, J

    2000-02-01

    Recombinant anti-testosterone wild-type Fab fragment and mutant Fab fragments with high binding selectivity developed by protein engineering have been crystallized with and without ligands. Crystals of these Fab fragments were obtained by the vapour-diffusion technique at room temperature using solutions of PEG 3350 with various biological buffers and with a wide pH range. So far, five data sets have been collected from crystals of three Fab-antigen complexes and from two uncomplexed Fab fragments, with resolutions ranging from 2.10 to 3.1 A. Crystallization conditions for Fab fragments were found by using modifications of the low ionic strength PEG 3350 series. Suitable concentrations of PEG 400, MPD and glycerol solutions for use as cryoprotectants in PEG 3350 solutions have been determined. One useful observation was that PEG 3350 is able to work alone as a cryoprotectant. The screening protocol used requires a smaller amount of protein material to achieve auspicious pre-crystals than previously. Results support the claim that PEG 3350 is more suitable for the crystallization of Fab fragments than higher molecular weight PEGs.

  12. Quantitative glycan profiling of normal human plasma derived immunoglobulin and its fragments Fab and Fc.

    Science.gov (United States)

    Anumula, Kalyan Rao

    2012-08-31

    Typical clinical grade human IgG (intravenous immunoglobulin, IVIG), used for carbohydrate analysis, is derived from thousands of healthy donors. Quantitative high-resolution glycan profiles of IgG and its Fc-Fab fragments are presented here. Glycan profiles were established following digestions with Fc specific endoglycosidase S and generic PNGase F under denaturing and non-denaturing (native) conditions. The native PNGase F glycan profile of IgG was similar (but not identical) to that of Endo S. Endo S profiles did not contain the glycans with bisecting GlcNAc. PNGase F glycan profiles were the same for Fc fragments that were isolated from pepsin and Ide S protease digests. Both isolated Fab fragments and the previously deglycosylated IVIG (native conditions) yielded the same glycan profile. Glycan profiles were established using high resolution HPLC with 2-aminobenzoic acid (2AA) labeling. An accurate determination of sialylation levels can be made by this method. Carbohydrate content in Fc and Fab was determined using an internal standard and corrected for both protein and glycan recoveries. Fab portion contained about 14% of the total carbohydrate which translates to 2.3 sugar chains per mol in IVIG where 2 chains are located in the CH2 domain of the Fc. Fc glycans consisted of neutral (N) 84.5%; mono-sialylated (S1) 15% and di-sialylated (S2) 0.5%. In contrast, Fab contained N, 21%; S1, 43% and S2, 36%. The distribution of bisecting N-acetylglucosamine and fucose was found to be very different in various glycans (N, S1 and S2) found in Fab and Fc. Total IgG glycan profile (Fab plus Fc) contained N, 78.5%; S1, 17% and S2, 4.5%. Percent distribution of glycans G0, G1 and G2 (with 0, 1 and 2 two galactoses) was 26, 49 and 25 respectively within the 78% of the neutral glycans. Glycan profiles were nearly the same for various clinical grade IVIG preparations from various manufacturers. A fast HPLC profiling method was developed for the separation and quantitation

  13. Crystallization and preliminary X-ray crystallographic analysis of enoyl-ACP reductase III (FabL) from Bacillus subtilis

    International Nuclear Information System (INIS)

    Kim, Kook-Han; Park, Joon Kyu; Ha, Byung Hak; Moon, Jin Ho; Kim, Eunice EunKyeong

    2007-01-01

    Enoyl-ACP reductase III (FabL) from B. subtilis has been overexpressed, purified and crystallized. The crystal belongs to space group P622, with unit-cell parameters a = b = 139.56, c = 62.75 Å, α = β = 90, γ = 120°, and data were collected to 2.5 Å resolution using synchrotron radiation. Enoyl-[acyl-carrier protein] reductase (enoyl-ACP reductase; ENR) is a key enzyme in type II fatty-acid synthase that catalyzes the last step in each elongation cycle. It has been considered as an antibiotic target since it is an essential enzyme in bacteria. However, recent studies indicate that some pathogens have more than one ENR. Bacillus subtilis is reported to have two ENRs, namely BsFabI and BsFabL. While BsFabI is similar to other FabIs, BsFabL shows very little sequence similarity and is NADPH-dependent instead of NADH-dependent as in the case of FabI. In order to understand these differences on a structural basis, BsFabL has been cloned, expressed and and crystallized. The crystal belongs to space group P622, with unit-cell parameters a = b = 139.56, c = 62.75 Å, α = β = 90, γ = 120° and one molecule of FabL in the asymmetric unit. Data were collected using synchrotron radiation (beamline 4A at the Pohang Light Source, Korea). The crystal diffracted to 2.5 Å resolution

  14. The Female Athlete Body (FAB) study: Rationale, design, and baseline characteristics.

    Science.gov (United States)

    Stewart, Tiffany M; Pollard, Tarryn; Hildebrandt, Tom; Beyl, Robbie; Wesley, Nicole; Kilpela, Lisa Smith; Becker, Carolyn Black

    2017-09-01

    Eating Disorders (EDs) are serious psychiatric illnesses marked by psychiatric comorbidity, medical complications, and functional impairment. Research indicates that female athletes are often at greater risk for developing ED pathology versus non-athlete females. The Female Athlete Body (FAB) study is a three-site, randomized controlled trial (RCT) designed to assess the efficacy of a behavioral ED prevention program for female collegiate athletes when implemented by community providers. This paper describes the design, intervention, and participant baseline characteristics. Future papers will discuss outcomes. Female collegiate athletes (N=481) aged 17-21 were randomized by site, team, and sport type to either FAB or a waitlist control group. FAB consisted of three sessions (1.3h each) of a behavioral ED prevention program. Assessments were conducted at baseline (pre-intervention), post-intervention (3weeks), and six-, 12-, and 18-month follow-ups. This study achieved 96% (N=481) of target recruitment (N=500). Few group differences emerged at baseline. Total sample analyses revealed moderately low baseline instances of ED symptoms and clinical cases. Health risks associated with EDs necessitate interventions for female athletes. The FAB study is the largest existing RCT for female athletes aimed at both reduction of ED risk factors and ED prevention. The methods presented and population recruited for this study represent an ideal intervention for assessing the effects of FAB on both the aforementioned outcomes. We anticipate that findings of this study (reported in future papers) will make a significant contribution to the ED risk factor reduction and prevention literature. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Native MS and ECD Characterization of a Fab-Antigen Complex May Facilitate Crystallization for X-ray Diffraction

    Science.gov (United States)

    Zhang, Ying; Cui, Weidong; Wecksler, Aaron T.; Zhang, Hao; Molina, Patricia; Deperalta, Galahad; Gross, Michael L.

    2016-07-01

    Native mass spectrometry (MS) and top-down electron-capture dissociation (ECD) combine as a powerful approach for characterizing large proteins and protein assemblies. Here, we report their use to study an antibody Fab (Fab-1)-VEGF complex in its near-native state. Native ESI with analysis by FTICR mass spectrometry confirms that VEGF is a dimer in solution and that its complex with Fab-1 has a binding stoichiometry of 2:2. Applying combinations of collisionally activated dissociation (CAD), ECD, and infrared multiphoton dissociation (IRMPD) allows identification of flexible regions of the complex, potentially serving as a guide for crystallization and X-ray diffraction analysis.

  16. [FAB immunoglobulin fragments. I. The comparative characteristics of the serological and virus-neutralizing properties of a gamma globulin against tick-borne encephalitis and of the FAB fragments isolated from it].

    Science.gov (United States)

    Barban, P S; Minaeva, V M; Pantiukhina, A N; Startseva, M G

    1976-06-01

    A comparative study was made of the serological properties and virus-neutralizing activity of antiencephalitis gamma-globulin and Fab-fragments isolated from it by gel-filtration. Horse immunoglobulins against the autumno-summer tick-borne encephalitis virus could be disintegrated with the aid of papaine to monovalent Fab-fragments which (according to the complement fixation reaction, the test of suppression of the complement fixation, and the HAIT) retained the serological activity whose level was compared with that of the serological activity of gamma-globulin. Fab-fragments possessed a marked virus-neutralizing activity. The mean value of a logarithm of the neutralization index was 2.65 +/- 0.2 for Fab-fragments and 3.74 +/- 0.38 for gamma-globulin (P less than 0.01).

  17. Generation of a haptoglobin-hemoglobin complex-specific Fab antibody blocking the binding of the complex to CD163

    DEFF Research Database (Denmark)

    Horn, Ivo R; Nielsen, Marianne Jensby; Madsen, Mette

    2003-01-01

    During intravascular hemolysis hemoglobin (Hb) binds to haptoglobin (Hp) leading to endocytosis of the complex by the macrophage receptor, CD163. In the present study, we used a phage-display Fab antibody strategy to explore if the complex formation between Hp and Hb leads to exposure of antigenic...... epitopes specific for the complex. By Hp-Hb-affinity screening of a phage-Fab library, we isolated a phage clone against the ligand complex. Surface plasmon resonance analyses of the Fab part expressed as a recombinant protein revealed a high affinity binding (KD = 3.9 nm) to Hp-Hb, whereas no binding...... was measured for non-complexed Hp or Hb. The Fab antibody completely inhibited the binding of 125I-labeled Hp-Hb complexes to CD163 and blocked their uptake in CD163-transfected cells. In conclusion, we have raised a receptor-blocking antibody specifically recognizing the Hp-Hb complex. In addition to provide...

  18. 188Re DD-3B6/22 Fab' for use in therapy of ovarian cancer: labelling and animal studies

    International Nuclear Information System (INIS)

    Schmidt, Peter F.; Smith, Suzanne V.; Bundesen, Peter G.

    1998-01-01

    A fast and high yielding method of 188 Re radiolabelling DD-3B6/22 Fab' is described. An inert atmosphere [N 2 (g)] and ascorbic acid was essential for preparation and storage of therapeutic levels (≤2 GBq/mg) for up to 24 h. Immunoreactivity was greater than 75%. Pharmacokinetic studies in nu/nu mice demonstrated localisation of 188 Re DD-3B6/22 Fab' was equivalent and correlated well with the behaviour observed for 99m Tc DD-3B6/22 Fab' used to image ovarian cancer. Excellent stability at the target site in vivo supports the potential use of 188 Re DD-3B6/22 Fab' in the therapy of ovarian cancer

  19. Application of a layout/material handling design method to a furnace area in a 300 mm wafer fab

    NARCIS (Netherlands)

    Hesen, P.M.C.; Renders, P.J.J.; Rooda, J.E.

    2001-01-01

    For many years, material handling within the semiconductor industry has become increasingly important. With the introduction of 300 mm wafer production, ergonomics and product safety become more critical. Therefore, the manufacturers of semiconductor wafer fabs are considering the automation of

  20. Construction and expression of a functional monoclonal antibody SZ-51 specific for GMP-140 chimeric fab fragment in Escherichia coli

    International Nuclear Information System (INIS)

    Gu Jianming; Zhang Xiaomin; Xia Lijun; Wan Haiying; Liu Yue; Li Peixia; Ruan Changgeng

    1996-04-01

    The variable region cDNAs of a monoclonal antibody SZ-51 specific for α-granule membrane protein (GMP-140) on the surface of activated human platelets were spliced with the constant region cDNA of the heavy chain CH1 and light chain k of human Ig G by means of the gene recombination techniques. The above recombinant gene was amplified by the polymerase chain reaction (PCR). The expression vector of phage plasmid pHEN1 SZ-51 Fab/Hu was constructed. The pHEN1-51 Fab/Hu was introduced into non-suppressor E. coli HB2151. The amount of expression of SZ-51 chimeric Fab/Hu measured by quantitative ELISA was about 500 μg/L. Western blot demonstrated that the SZ-51 chimeric Fab fragment could specifically bind to GMP-140. (2 figs.)

  1. Plants of an Eucalyptus clone damaged by Scolytidae and Platypodidae (Coleoptera Plantas de Eucalyptus atacadas por Scolytidae e Platypodidae (Coleoptera

    Directory of Open Access Journals (Sweden)

    José Cola Zanuncio

    2005-05-01

    Full Text Available Species of Scolytidae and Platypodidae were evaluated in a plantation using a clone of Eucalyptus grandis x Eucalyptus urophylla in the State of Minas Gerais, Brazil. Wood disks with galleries were taken from 15 eucalyptus trees between 0.5 and 1.5 m height. After collection, all trees attacked were burned. Woodborers obtained from these disks were identified as Premnobius cavipennis, Premnobius ambitiosus, Dryocoetoides cristatus (Coleoptera: Scolytidae and Euplatypus parallelus (Coleoptera: Platypodidae. This is the first report of D. cristatus, a Brazilian native species, damaging eucalyptus, which shows its adaptation to this plant.O objetivo deste trabalho foi avaliar o ataque de Scolytidae e Platypodidae em talhão de clone Eucalyptus grandis x Eucalyptus urophylla na região de Montes Claros, Estado de Minas Gerais, Brasil. Discos de madeira com galerias desses insetos foram retirados de 15 árvores de eucalipto entre 0,5 e 1,5 m de altura. Após a coleta deste material, as árvores atacadas foram queimadas. Os indivíduos da ordem Coleoptera obtidos desses discos foram identificados como Premnobius cavipennis, Premnobius ambitiosus e Dryocoetoides cristatus (Coleoptera: Scolytidae e Euplatypus parallelus (Coleoptera: Platypodidae. O ataque de D. cristatus, espécie nativa do Brasil, ao eucalipto ainda não havia sido relatado, o que mostra sua adaptação a essa planta.

  2. A novel human Fab antibody for Trop2 inhibits breast cancer growth in vitro and in vivo.

    Science.gov (United States)

    Lin, Hong; Zhang, Huiling; Wang, Jun; Lu, Meiping; Zheng, Feng; Wang, Changjun; Tang, Xiaojun; Xu, Ning; Chen, Renjie; Zhang, Dawei; Zhao, Ping; Zhu, Jin; Mao, Yuan; Feng, Zhenqing

    2014-03-01

    Human trophoblastic cell surface antigen 2 (Trop2) has been suggested as an oncogene, which is associated with the different types of tumors. In this study, a human Fab antibody against Trop2 extracellular domain was isolated from phage library by phage display technology, and characterized by ELISA, FACS, fluorescence staining and Western blotting analysis. MTT, apoptosis assay and wound healing assay were employed to evaluate the inhibitory effects of Trop2 Fab on breast cancer cell growth in vitro, while tumor-xenograft model was employed to evaluate the inhibitory effects on breast cancer growth in vivo. The results showed that Trop2 Fab inhibited the proliferation, induced the apoptosis and suspended the migration of MDA-MB-231 cells in a dose dependent manner. The expression caspase-3 was activated, and the expression of Bcl-2 was reduced while that of Bax was elevated in MDA-MB-231 cells by treating with Trop2 Fab. In addition, Trop2 Fab inhibited the growth of breast cancer xenografts and the expression of Bcl-2 was reduced while that of Bax was elevated in xenografts. Trop2 Fab, which was isolated successfully in this research, is a promising therapeutic agent for the treatment of Trop2 expressing breast cancer. © 2013 UICC.

  3. Improving Assessment of Work Related Mental Health Function Using the Work Disability Functional Assessment Battery (WD-FAB).

    Science.gov (United States)

    Marfeo, Elizabeth E; Ni, Pengsheng; McDonough, Christine; Peterik, Kara; Marino, Molly; Meterko, Mark; Rasch, Elizabeth K; Chan, Leighton; Brandt, Diane; Jette, Alan M

    2018-03-01

    Purpose To improve the mental health component of the Work Disability Functional Assessment Battery (WD-FAB), developed for the US Social Security Administration's (SSA) disability determination process. Specifically our goal was to expand the WD-FAB scales of mood & emotions, resilience, social interactions, and behavioral control to improve the depth and breadth of the current scales and expand the content coverage to include aspects of cognition & communication function. Methods Data were collected from a random, stratified sample of 1695 claimants applying for the SSA work disability benefits, and a general population sample of 2025 working age adults. 169 new items were developed to replenish the WD-FAB scales and analyzed using factor analysis and item response theory (IRT) analysis to construct unidimensional scales. We conducted computer adaptive test (CAT) simulations to examine the psychometric properties of the WD-FAB. Results Analyses supported the inclusion of four mental health subdomains: Cognition & Communication (68 items), Self-Regulation (34 items), Resilience & Sociability (29 items) and Mood & Emotions (34 items). All scales yielded acceptable psychometric properties. Conclusions IRT methods were effective in expanding the WD-FAB to assess mental health function. The WD-FAB has the potential to enhance work disability assessment both within the context of the SSA disability programs as well as other clinical and vocational rehabilitation settings.

  4. Structural characterization of POM6 Fab and mouse prion protein complex identifies key regions for prions conformational conversion.

    Science.gov (United States)

    Baral, Pravas Kumar; Swayampakula, Mridula; Aguzzi, Adriano; James, Michael N G

    2018-05-01

    Conversion of the cellular prion protein PrP C into its pathogenic isoform PrP S c is the hallmark of prion diseases, fatal neurodegenerative diseases affecting many mammalian species including humans. Anti-prion monoclonal antibodies can arrest the progression of prion diseases by stabilizing the cellular form of the prion protein. Here, we present the crystal structure of the POM6 Fab fragment, in complex with the mouse prion protein (moPrP). The prion epitope of POM6 is in close proximity to the epitope recognized by the purportedly toxic antibody fragment, POM1 Fab also complexed with moPrP. The POM6 Fab recognizes a larger binding interface indicating a likely stronger binding compared to POM1. POM6 and POM1 exhibit distinct biological responses. Structural comparisons of the bound mouse prion proteins from the POM6 Fab:moPrP and POM1 Fab:moPrP complexes reveal several key regions of the prion protein that might be involved in initiating mis-folding events. The structural data of moPrP:POM6 Fab complex are available in the PDB under the accession number www.rcsb.org/pdb/search/structidSearch.do?structureId=6AQ7. © 2018 Federation of European Biochemical Societies.

  5. Genetic variability and resistance of cultivars of cowpea [Vigna unguiculata (L.) Walp] to cowpea weevil (Callosobruchus maculatus Fabr.).

    Science.gov (United States)

    Vila Nova, M X; Leite, N G A; Houllou, L M; Medeiros, L V; Lira Neto, A C; Hsie, B S; Borges-Paluch, L R; Santos, B S; Araujo, C S F; Rocha, A A; Costa, A F

    2014-03-31

    The cowpea weevil (Callosobruchus maculatus Fabr.) is the most destructive pest of the cowpea bean; it reduces seed quality. To control this pest, resistance testing combined with genetic analysis using molecular markers has been widely applied in research. Among the markers that show reliable results, the inter-simple sequence repeats (ISSRs) (microsatellites) are noteworthy. This study was performed to evaluate the resistance of 27 cultivars of cowpea bean to cowpea weevil. We tested the resistance related to the genetic variability of these cultivars using ISSR markers. To analyze the resistance of cultivars to weevil, a completely randomized test design with 4 replicates and 27 treatments was adopted. Five pairs of the insect were placed in 30 grains per replicate. Analysis of variance showed that the number of eggs and emerged insects were significantly different in the treatments, and the means were compared by statistical tests. The analysis of the large genetic variability in all cultivars resulted in the formation of different groups. The test of resistance showed that the cultivar Inhuma was the most sensitive to both number of eggs and number of emerged adults, while the TE96-290-12-G and MNC99-537-F4 (BRS Tumucumaque) cultivars were the least sensitive to the number of eggs and the number of emerged insects, respectively.

  6. Salinity-dependent nickel accumulation and effects on respiration, ion regulation and oxidative stress in the galaxiid fish, Galaxias maculatus.

    Science.gov (United States)

    Blewett, Tamzin A; Wood, Chris M; Glover, Chris N

    2016-07-01

    Inanga (Galaxias maculatus) are a euryhaline and amphidromous Southern hemisphere fish species inhabiting waters highly contaminated in trace elements such as nickel (Ni). Ni is known to exert its toxic effects on aquatic biota via three key mechanisms: inhibition of respiration, impaired ion regulation, and stimulation of oxidative stress. Inanga acclimated to freshwater (FW), 50% seawater (SW) or 100% SW were exposed to 0, 150 or 2000 μg Ni L(-1), and tissue Ni accumulation, metabolic rate, ion regulation (tissue ions, calcium (Ca) ion influx), and oxidative stress (catalase activity, protein carbonylation) were measured after 96 h. Ni accumulation increased with Ni exposure concentration in gill, gut and remaining body, but not in liver. Only in the gill was Ni accumulation affected by exposure salinity, with lower branchial Ni burdens in 100% and 50% SW inanga, relative to FW fish. There were no Ni-dependent effects on respiration, or Ca influx, and the only Ni-dependent effect on tissue ion content was on gill potassium. Catalase activity and protein carbonylation were affected by Ni, primarily in FW, but only at 150 μg Ni L(-1). Salinity therefore offsets the effects of Ni, despite minimal changes in Ni bioavailability. These data suggest only minor effects of Ni in inanga, even at highly elevated environmental Ni concentrations. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. In-Situ Ion Source Cleaning: Review of Chemical Mechanisms and Evaluation Data at Production Fabs

    International Nuclear Information System (INIS)

    Kaim, R.; Bishop, S.; Byl, O.; Eldridge, D.; Marganski, P.; Mayer, J.; Sweeney, J.; Yedave, S.; Fuchs, D.; Spreitzer, S.; Vogel, J.; Dunn, J.; Lundquist, P.; Rolland, J.; Romig, T.; Newman, D.; Mitchell, M.; Ditzler, K.

    2008-01-01

    Since the concept of chemical in-situ ion implanter cleaning was introduced at IIT2006 [1], evaluations of the XeF 2 cleaning technology have taken place or are ongoing at more than 40 production fabs worldwide. Testing has been focused on assessing effects of cleaning in the source arc chamber and extraction regions. In this paper we describe use of the cleaning technology in a production environment and summarize evaluation data showing advantages of the technology for improving ion source life, reducing glitching, improving beam auto-tuning and avoiding species cross-contamination. More details of the evaluations are given in several separate papers submitted to this Conference. We have supplemented the fab production data with laboratory experiments designed to investigate the reactivity of XeF 2 and fundamental aspects of the source deposition and cleaning processes. These experiments are summarized here, and more details can be found in separate papers submitted to this Conference

  8. Characterization of a recombinant humanized anti-cocaine monoclonal antibody and its Fab fragment.

    Science.gov (United States)

    Kirley, Terence L; Norman, Andrew B

    2015-01-01

    Variations of post-translational modifications are important for stability and in vivo behavior of therapeutic antibodies. A recombinant humanized anti-cocaine monoclonal antibody (h2E2) was characterized for heterogeneity of N-linked glycosylation and disulfide bonds. In addition, charge heterogeneity, which is partially due to the presence or absence of C-terminal lysine on the heavy chains, was examined. For cocaine overdose therapy, Fab fragments may be therapeutic, and thus, a simplified method of generation, purification, and characterization of the Fab fragment generated by Endoproteinase Lys-C digestion was devised. Both the intact h2E2 antibody and purified Fab fragments were analyzed for their affinities for cocaine and 2 of its metabolites, benzoylecgonine and cocaethylene, by fluorescence quenching of intrinsic antibody tyrosine and tryptophan fluorescence resulting from binding of these drugs. Binding constants obtained from fluorescence quenching measurements are in agreement with recently published radioligand and ELISA binding assays. The dissociation constants determined for the h2E2 monoclonal and its Fab fragment are approximately 1, 5, and 20 nM for cocaethylene, cocaine, and benzoylecgonine, respectively. Tryptophan fluorescence quenching (emission at 330 nm) was measured after either excitation of tyrosine and tryptophan (280 nm) or selective excitation of tryptophan alone (295 nm). More accurate binding constants are obtained using tryptophan selective excitation at 295 nm, likely due to interfering absorption of cocaine and metabolites at 280 nm. These quenching results are consistent with multiple tryptophan and tyrosine residues in or near the predicted binding location of cocaine in a previously published 3-D model of this antibody's variable region.

  9. Fab the coming revolution on your desktop : from personal computers to personal fabrication

    CERN Document Server

    Gershenfeld, Neil

    2005-01-01

    What if you could someday put the manufacturing power of an automobile plant on your desktop? According to Neil Gershenfeld, the renowned MIT scientist and inventor, the next big thing is personal fabrication-the ability to design and produce your own products, in your own home, with a machine that combines consumer electronics and industrial tools. Personal fabricators are about to revolutionize the world just as personal computers did a generation ago, and Fab shows us how.

  10. Efficient production of antibody Fab fragment by transient gene expression in insect cells.

    Science.gov (United States)

    Mori, Keita; Hamada, Hirotsugu; Ogawa, Takafumi; Ohmuro-Matsuyama, Yuki; Katsuda, Tomohisa; Yamaji, Hideki

    2017-08-01

    Transient gene expression allows a rapid production of diverse recombinant proteins in early-stage preclinical and clinical developments of biologics. Insect cells have proven to be an excellent platform for the production of functional recombinant proteins. In the present study, the production of an antibody Fab fragment by transient gene expression in lepidopteran insect cells was investigated. The DNA fragments encoding heavy-chain (Hc; Fd fragment) and light-chain (Lc) genes of an Fab fragment were individually cloned into the plasmid vector pIHAneo, which contained the Bombyx mori actin promoter downstream of the B. mori nucleopolyhedrovirus (BmNPV) IE-1 transactivator and the BmNPV HR3 enhancer for high-level expression. Trichoplusia ni BTI-TN-5B1-4 (High Five) cells were co-transfected with the resultant plasmid vectors using linear polyethyleneimine. When the transfection efficiency was evaluated, a plasmid vector encoding an enhanced green fluorescent protein (EGFP) gene was also co-transfected. Transfection and culture conditions were optimized based on both the flow cytometry of the EGFP expression in transfected cells and the yield of the secreted Fab fragments determined by enzyme-linked immunosorbent assay (ELISA). Under optimal conditions, a yield of approximately 120 mg/L of Fab fragments was achieved in 5 days in a shake-flask culture. Transient gene expression in insect cells may offer a promising approach to the high-throughput production of recombinant proteins. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  11. Mass spectra of alkaline earth salts with a FAB source. Complexation with crown ethers

    International Nuclear Information System (INIS)

    Ulrich, J.

    1987-01-01

    With a liquid desorption FAB source it is possible to obtain alkaline earth metal ions complexed by a crown ether. Conditions for formation of these complexes ions are examined for selection of the complexing agent in function of cation size. Behaviour of alkaline and alkaline earth compounds are compared allowing the differentiation of ion extraction phenomena by liquid desorption ion source and solvent extraction [fr

  12. Anticomplementary Activity of Horse IgG and F(ab')2 Antivenoms

    OpenAIRE

    Squaiella-Baptistão, Carla Cristina; Marcelino, José Roberto; Ribeiro da Cunha, Luiz Eduardo; Gutiérrez, José María; Tambourgi, Denise V.

    2014-01-01

    2094-01 Embargo por política editorial Envenomation by poisonous animals is a neglected condition according to the World Health Organization (WHO). Antivenoms are included in the WHO Essential Medicines List. It has been assumed that immunoglobulin G (IgG) antivenoms could activate the complement system through Fc and induce early adverse reactions (EARs). However, data in the literature indicate that F(ab')2 fragments can also activate the complement system. Herein, we show that several b...

  13. Production and purification of polyclonal antibody against F(ab')2 fragment of human immunoglobulin G

    OpenAIRE

    Nasiri, Hadi; Valedkarimi, Zahra; Aghebati-Maleki, Leili; Abdolalizadeh, Jalal; Kazemi, Tohid; Esparvarinha, Mojghan; Majidi, Jafar

    2017-01-01

    Antibodies are essential tools of biomedical and biochemical researches. Polyclonal antibodies are produced against different epitopes of antigens. Purified F(ab')2 can be used for animal’s immunization to produce polyclonal antibodies. Human immunoglobulin G (IgG) was purified by ion exchange chromatography method. In all stages verification method of the purified antibodies was sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purified IgG was digested by pepsin enzyme a...

  14. Aerial image measurement technique for automated reticle defect disposition (ARDD) in wafer fabs

    Science.gov (United States)

    Zibold, Axel M.; Schmid, Rainer M.; Stegemann, B.; Scheruebl, Thomas; Harnisch, Wolfgang; Kobiyama, Yuji

    2004-08-01

    The Aerial Image Measurement System (AIMS)* for 193 nm lithography emulation has been brought into operation successfully worldwide. A second generation system comprising 193 nm AIMS capability, mini-environment and SMIF, the AIMS fab 193 plus is currently introduced into the market. By adjustment of numerical aperture (NA), illumination type and partial illumination coherence to match the conditions in 193 nm steppers or scanners, it can emulate the exposure tool for any type of reticles like binary, OPC and PSM down to the 65 nm node. The system allows a rapid prediction of wafer printability of defects or defect repairs, and critical features, like dense patterns or contacts on the masks without the need to perform expensive image qualification consisting of test wafer exposures followed by SEM measurements. Therefore, AIMS is a mask quality verification standard for high-end photo masks and established in mask shops worldwide. The progress on the AIMS technology described in this paper will highlight that besides mask shops there will be a very beneficial use of the AIMS in the wafer fab and we propose an Automated Reticle Defect Disposition (ARDD) process. With smaller nodes, where design rules are 65 nm or less, it is expected that smaller defects on reticles will occur in increasing numbers in the wafer fab. These smaller mask defects will matter more and more and become a serious yield limiting factor. With increasing mask prices and increasing number of defects and severability on reticles it will become cost beneficial to perform defect disposition on the reticles in wafer production. Currently ongoing studies demonstrate AIMS benefits for wafer fab applications. An outlook will be given for extension of 193 nm aerial imaging down to the 45 nm node based on emulation of immersion scanners.

  15. Nebulized Anti-IL-13 Monoclonal Antibody Fab' Fragment Reduces Allergen-Induced Asthma

    OpenAIRE

    Hacha, Jonathan; Tomlinson, K; Maertens, Ludovic; Paulissen, Geneviève; Rocks, Natacha; Foidart, Jean-Michel; Noël, Agnès; Palframan, R; Guéders, Maud; Cataldo, Didier

    2012-01-01

    Rationale: Interleukin-13 (IL-13) is a prototypic Th2 cytokine and a central mediator of the complex cascade of events leading to asthmatic phenotype. Indeed, IL-13 plays key roles in IgE synthesis, bronchial hyperresponsiveness, mucus hypersecretion, subepithelial fibrosis and eosinophil infiltration. Objectives: We assessed the potential efficacy of inhaled anti-IL-13 monoclonal antibody Fab' fragment on allergen-induced airway inflammation, hyperresponsiveness and remodeling in an experime...

  16. Molecular markers detect cryptic predation on coffee berry borer (Coleoptera: Curculionidae) by silvanid and laemophloeid flat bark beetles (Coleoptera: Silvanidae, Laemophloeidae) in coffee beans

    Science.gov (United States)

    The coffee berry borer, Hypothenemus hampei(Coleoptera: Curculionidae)(Ferrari), is a serious pest of coffee worldwide and has been recently introduced in Hawai’i, first detected in the state in 2010. Adult silvanid flat bark beetles, Cathartus quadricollis (Coleoptera: Silvanidae) and adult laemoph...

  17. Severe adverse drug reaction following Crotalidae Polyvalent Immune Fab (Ovine) administration for copperhead snakebite.

    Science.gov (United States)

    Lepak, Maryjoy R; Bochenek, Samantha H; Bush, Sean P

    2015-01-01

    To present the case of a severe anaphylactic/anaphylactoid reaction to Crotalidae Polyvalent Immune Fab (Ovine) in a patient bitten by a copperhead snake. A 68-year-old man presented with progressive envenomation after receiving a copperhead snakebite on each hand. Crotalinae Fab antivenom was administered. While the initial and only dose was partially infusing, the patient developed an adverse drug reaction (ADR) of urticaria and hypotension, which resolved with cessation of the infusion, recurred with resumption of the infusion, and ultimately was completed with supportive care. An additional episode of hypotension, urticaria, and angioedema occurred shortly after antivenom therapy completion. Epinephrine was administered, resolving the reaction with complete patient recovery. The event received a Naranjo score of 10, indicating a definite ADR. Treating copperhead snakebites with antivenom is a matter of debate. Concern over adverse events and cost induce some physicians to manage copperhead bites without antivenom because they are generally milder in severity. As demonstrated in this case, severe ADR can occur with Crotalinae Fab antivenom, and its efficacy for copperhead envenoming needs to be better established via placebo-controlled, randomized trials. © The Author(s) 2014.

  18. Acute myocardial infarct imaging with indium-111-labeled monoclonal antimyosin Fab

    International Nuclear Information System (INIS)

    Khaw, B.A.; Yasuda, T.; Gold, H.K.; Leinbach, R.C.; Johns, J.A.; Kanke, M.; Barlai-Kovach, M.; Strauss, H.W.; Haber, E.

    1987-01-01

    Indium-111 monoclonal antimyosin Fab scintigraphy was used to detect myocardial necrosis in 52 of 54 patients (96.3%) with acute myocardial infarction. Infarcts were visualized when coronary arteries were persistently occluded (n = 10), became patent after thrombolysis (n = 33), or became patent after spontaneous reperfusion (n = 7). Posteroinferolateral visualizations were obtained in two patients with clinical and enzymatic evidence of infarction but normal electrocardiograms. Of the two patients in whom no infarcts were visualized, one had an anterior myocardial infarct. This patient underwent successful thrombolytic therapy, with attendant minimization of creatine kinase release. The other patient had a small, nonreperfused inferior myocardial infarct. Five patients with a history of remote infarction and acute necrosis showed antimyosin uptake only in regions concordant with the acute episodes of infarction, and radiolabeled antimyosin Fab localized in neither old infarcts nor normal, noninfarcted myocardium. Antimyosin Fab scintigraphy, thus, appears to be a highly specific means of delineating necrotic myocardium, at least in this limited and selected group of patients

  19. PET imaging of osteosarcoma in dogs using a fluorine-18-labeled monoclonal antibody fab fragment

    International Nuclear Information System (INIS)

    Page, R.L.; Garg, P.K.; Gard, S.

    1994-01-01

    Four dogs with histologically confirmed osteogenic sarcoma were studied with PET following intravenous injection of the 18 F-labeled Fab fragment of TP-3, a monoclonal antibody specific for human and canine osteosarcomas. The antibody fragment was labeled using the N-succinimidyl (8-(4'-( 18 F)fluorobenzyl)amino)suberate acylation agent. Blood clearance of activity was biphasic in all dogs but half-times were variable (T 1/2β = 2-13 hr). Catabolism of labeled Fab was reflected by the decrease in protein-associated activity in serum from more than 90% at 1 min to 60%-80% at 4 hr. PET images demonstrated increased accumulation of 18 F at the primary tumor site relative to normal contralateral bone in one dog as early as 15 min after injection. Biopsies obtained after euthanasia indicated higher uptake at the edges of the tumor as observed on the PET scans. Tumor uptake was 1-3 x 10 -3 % injected dose/g, a level similar to that reported for other Fab fragments in human tumors. In the three dogs with metastatic disease, early PET images reflected activity in the blood pool but later uptake was observed in suspected metastatic sites. These results, although preliminary, suggest that PET imaging of 18 F-labeled antibody fragments is feasible and that dogs with spontaneous tumors could be a valuable model for preclinical research with radioimmunoconjugates. 34 refs., 6 figs., 2 tabs

  20. PET imaging of osteosarcoma in dogs using a fluorine-18-labeled monoclonal antibody fab fragment

    Energy Technology Data Exchange (ETDEWEB)

    Page, R.L.; Garg, P.K.; Gard, S. [North Carolina State Univ., Raleigh, NC (United States)]|[Duke Univ. Medical Center, Durham, NC (United States)]|[North Carolina and Norke Radium Hospital, Oslo (Norway)] [and others

    1994-09-01

    Four dogs with histologically confirmed osteogenic sarcoma were studied with PET following intravenous injection of the {sup 18}F-labeled Fab fragment of TP-3, a monoclonal antibody specific for human and canine osteosarcomas. The antibody fragment was labeled using the N-succinimidyl (8-(4{prime}-({sup 18}F)fluorobenzyl)amino)suberate acylation agent. Blood clearance of activity was biphasic in all dogs but half-times were variable (T{sub 1/2{beta}} = 2-13 hr). Catabolism of labeled Fab was reflected by the decrease in protein-associated activity in serum from more than 90% at 1 min to 60%-80% at 4 hr. PET images demonstrated increased accumulation of {sup 18}F at the primary tumor site relative to normal contralateral bone in one dog as early as 15 min after injection. Biopsies obtained after euthanasia indicated higher uptake at the edges of the tumor as observed on the PET scans. Tumor uptake was 1-3 x 10{sup -3}% injected dose/g, a level similar to that reported for other Fab fragments in human tumors. In the three dogs with metastatic disease, early PET images reflected activity in the blood pool but later uptake was observed in suspected metastatic sites. These results, although preliminary, suggest that PET imaging of {sup 18}F-labeled antibody fragments is feasible and that dogs with spontaneous tumors could be a valuable model for preclinical research with radioimmunoconjugates. 34 refs., 6 figs., 2 tabs.

  1. Regulation of fatty acid biosynthesis by the global regulator CcpA and the local regulator FabT in Streptococcus mutans

    OpenAIRE

    Faustoferri, R.C.; Hubbard, C.J.; Santiago, B.; Buckley, A.A.; Seifert, T.B.; Quivey, R.G.

    2014-01-01

    SMU.1745c, encoding a putative transcriptional regulator of the MarR family, maps to a location proximal to the fab gene cluster in Streptococcus mutans. Deletion of the SMU.1745c (fabTSm) coding region resulted in a membrane fatty acid composition comprised of longer-chained, unsaturated fatty acids (UFA), compared with the parent strain. Previous reports have indicated a role for FabT in regulation of genes in the fab gene cluster in other organisms, through binding to a palindromic DNA seq...

  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. Oriented Immobilization of Fab Fragments by Site-Specific Biotinylation at the Conserved Nucleotide Binding Site for Enhanced Antigen Detection.

    Science.gov (United States)

    Mustafaoglu, Nur; Alves, Nathan J; Bilgicer, Basar

    2015-09-08

    Oriented immobilization of antibodies and antibody fragments has become increasingly important as a result of the efforts to reduce the size of diagnostic and sensor devices to miniaturized dimensions for improved accessibility to the end-user. Reduced dimensions of sensor devices necessitate the immobilized antibodies to conserve their antigen binding activity for proper operation. Fab fragments are becoming more commonly used in small-scaled diagnostic devices due to their small size and ease of manufacture. In this study, we used the previously described UV-NBS(Biotin) method to functionalize Fab fragments with IBA-EG11-Biotin linker utilizing UV energy to initiate a photo-cross-linking reaction between the nucleotide binding site (NBS) on the Fab fragment and IBA-Biotin molecule. Our results demonstrate that immobilization of biotinylated Fab fragments via UV-NBS(Biotin) method generated the highest level of immobilized Fab on surfaces when compared to other typical immobilization methods while preserving antigen binding activity. UV-NBS(Biotin) method provided 432-fold, 114-fold, and 29-fold improved antigen detection sensitivity than physical adsorption, NHS-Biotin, and ε-NH3(+), methods, respectively. Additionally, the limit of detection (LOD) for PSA utilizing Fab fragments immobilized via UV-NBS(Biotin) method was significantly lower than that of the other immobilization methods, with an LOD of 0.4 pM PSA. In summary, site-specific biotinylation of Fab fragments without structural damage or loss in antigen binding activity provides a wide range of application potential for UV-NBS immobilization technique across numerous diagnostic devices and nanotechnologies.

  4. Pre- and Posttransplant IgA Anti-Fab Antibodies to Predict Long-term Kidney Graft Survival.

    Science.gov (United States)

    Amirzargar, M A; Amirzargar, A; Basiri, A; Hajilooi, M; Roshanaei, G; Rajabi, G; Solgi, G

    2015-05-01

    Immunologic factors are reliable markers for allograft monitoring, because of their seminal role in rejection process. One of these factors is the immunoglobulin (Ig)A anti-Fab of the IgG antibody. This study aimed to evaluate the predictive value of pre- and posttransplant levels of this marker for kidney allograft function and survival. Sera samples of 59 living unrelated donor kidney recipients were collected before and after transplantation (days 7, 14, and 30) and investigated for IgA anti-Fab of IgG antibody levels using enzyme-linked immunosorbent assay in relation with allograft outcome. Among 59 patients, 15 cases (25%) including 10 with acute rejection and 5 with chronic rejection episodes showed graft failure during a mean of 5 years of follow-up. High posttransplant levels of IgA anti-Fab antibodies were observed more frequently in patients with stable graft function (SGF) compared with patients with graft failure (P = 2 × 10(-6)). None of patients with acute or chronic rejection episodes had high levels of IgA anti-Fab antibodies at day 30 posttransplant compared with the SGF group (P = 10(-6) and P = .01, respectively). In addition, high levels of IgA anti-Fab antibody correlated with lesser concentration of serum creatinine at 1 month posttransplantation (P = .01). Five-year graft survival was associated with high levels of pre- and posttransplant IgA anti-Fab antibodies (P = .02 and P = .003, respectively). Our findings indicate the protective effect of higher levels of IgA anti-Fab antibodies regarding to kidney allograft outcomes and long-term graft survival. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Development of a high yielding E. coli periplasmic expression system for the production of humanized Fab' fragments.

    Science.gov (United States)

    Ellis, Mark; Patel, Pareshkumar; Edon, Marjory; Ramage, Walter; Dickinson, Robert; Humphreys, David P

    2017-01-01

    Humanized Fab' fragments may be produced in the periplasm of Escherichia coli but can be subject to degradation by host cell proteases. In order to increase Fab' yield and reduce proteolysis we developed periplasmic protease deficient strains of E. coli. These strains lacked the protease activity of Tsp, protease III and DegP. High cell density fermentations indicated Tsp deficient strains increased productivity two fold but this increase was accompanied by premature cell lysis soon after the induction of Fab' expression. To overcome the reduction in cell viability we introduced suppressor mutations into the spr gene. The mutations partially restored the wild type phenotype of the cells. Furthermore, we coexpressed a range of periplasmic chaperone proteins with the Fab', DsbC had the most significant impact, increasing humanized Fab' production during high cell density fermentation. When DsbC coexpression was combined with a Tsp deficient spr strain we observed an increase in yield and essentially restored "wild type" cell viability. We achieved a final periplasmic yield of over 2.4g/L (final cell density OD 600 105), 40 h post Fab' induction with minimal cell lysis.The data suggests that proteolysis, periplasm integrity, protein folding and disulphide bond formation are all potential limiting steps in the production of Fab' fragments in the periplasm of E. coli. In this body of work, we have addressed these limiting steps by utilizing stabilized protease deficient strains and chaperone coexpression. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:212-220, 2017. © 2016 American Institute of Chemical Engineers.

  6. Acute adverse events associated with the administration of Crotalidae polyvalent immune Fab antivenom within the North American Snakebite Registry.

    Science.gov (United States)

    Kleinschmidt, Kurt; Ruha, Anne-Michelle; Campleman, Sharan; Brent, Jeffrey; Wax, Paul

    2018-04-24

    Crotalidae Polyvalent Immune Fab (Fab Antivenom) is the primary Viperid antivenom used in the United States since 2000. Adverse event data associated with its use are limited. The purpose of this study is to describe the prevalence of acute adverse events associated with the use of Fab antivenom. The American College of Medical Toxicology's Toxicology Investigators Consortium maintains a prospective case registry of poisoned and envenomated patients managed by medical toxicologists at the bedside. This registry includes the North American Snakebite sub-registry. We performed a review of 438 cases entered into the Snakebite sub-registry. A total of 373 (85.2%) received at least one vial of Fab Antivenom. Forty percent were children. Adverse events occurred in 10 patients (2.7%) of whom six were adults. Rash was the most common adverse event. More severe adverse events (hypotension, bronchospasm, and/or angioedema) occurred in four (1.1%) patients. Prophylaxis was administered prior to Fab antivenom in 4.0%. Eight patients received various treatments for their adverse events. Neither the initial number of Fab antivenom vials, atopic history, nor prior envenomation correlated with the prevalence of adverse events. This prevalence of adverse events was lower than in previous studies and in a meta-analysis of 11 studies. The types of adverse events and treatments used are consistent with those in previous reports. There were no prior reports of prophylaxis use with which to compare. The prevalence of Fab antivenom adverse events in the North American Snakebite Registry was 2.7%.

  7. Functional Dissection of the Blocking and Bypass Activities of the Fab-8 Boundary in the Drosophila Bithorax Complex.

    Science.gov (United States)

    Kyrchanova, Olga; Mogila, Vladic; Wolle, Daniel; Deshpande, Girish; Parshikov, Alexander; Cléard, Fabienne; Karch, Francois; Schedl, Paul; Georgiev, Pavel

    2016-07-01

    Functionally autonomous regulatory domains direct the parasegment-specific expression of the Drosophila Bithorax complex (BX-C) homeotic genes. Autonomy is conferred by boundary/insulator elements that separate each regulatory domain from its neighbors. For six of the nine parasegment (PS) regulatory domains in the complex, at least one boundary is located between the domain and its target homeotic gene. Consequently, BX-C boundaries must not only block adventitious interactions between neighboring regulatory domains, but also be permissive (bypass) for regulatory interactions between the domains and their gene targets. To elucidate how the BX-C boundaries combine these two contradictory activities, we have used a boundary replacement strategy. We show that a 337 bp fragment spanning the Fab-8 boundary nuclease hypersensitive site and lacking all but 83 bp of the 625 bp Fab-8 PTS (promoter targeting sequence) fully rescues a Fab-7 deletion. It blocks crosstalk between the iab-6 and iab-7 regulatory domains, and has bypass activity that enables the two downstream domains, iab-5 and iab-6, to regulate Abdominal-B (Abd-B) transcription in spite of two intervening boundary elements. Fab-8 has two dCTCF sites and we show that they are necessary both for blocking and bypass activity. However, CTCF sites on their own are not sufficient for bypass. While multimerized dCTCF (or Su(Hw)) sites have blocking activity, they fail to support bypass. Moreover, this bypass defect is not rescued by the full length PTS. Finally, we show that orientation is critical for the proper functioning the Fab-8 replacement. Though the inverted Fab-8 boundary still blocks crosstalk, it disrupts the topology of the Abd-B regulatory domains and does not support bypass. Importantly, altering the orientation of the Fab-8 dCTCF sites is not sufficient to disrupt bypass, indicating that orientation dependence is conferred by other factors.

  8. Targeting human prostate cancer with 111In-labeled D2B IgG, F(ab')2 and Fab fragments in nude mice with PSMA-expressing xenografts.

    Science.gov (United States)

    Lütje, Susanne; van Rij, Catharina M; Franssen, Gerben M; Fracasso, Giulio; Helfrich, Wijnand; Eek, Annemarie; Oyen, Wim J; Colombatti, Marco; Boerman, Otto C

    2015-01-01

    D2B is a new monoclonal antibody directed against an extracellular domain of prostate-specific membrane antigen (PSMA), which is overexpressed in prostate cancer. The potential of D2B IgG, and F(ab')2 and Fab fragments of this antibody for targeting prostate cancer was determined in mice bearing subcutaneous prostate cancer xenografts. The optimal time point for imaging was determined in biodistribution and microSPECT imaging studies with (111)In-D2B IgG, (111)In-capromab pendetide, (111)In-D2B F(ab')2 and (111)In-D2B Fab fragments in mice with PSMA-expressing LNCaP and PSMA-negative PC3 tumors at several time points after injection. All (111)In-labeled antibody formats specifically accumulated in the LNCaP tumors, with highest uptake of (111)In-D2B IgG and (111)In-capromab pendetide at 168 h p.i. (94.8 ± 19.2% injected dose per gram (ID/g) and 16.7 ± 2.2% ID/g, respectively), whereas uptake of (111)In-D2B F(ab')2 and (111)In-D2B Fab fragments peaked at 24 h p.i. (12.1 ± 3.0% ID/g and 15.1 ± 2.9% ID/g, respectively). Maximum LNCaP tumor-to-blood ratios were 13.0 ± 2.3 (168 h p.i.), 6.2 ± 0.7 (24 h p.i.), 23.0 ± 4.0 (24 h p.i.) and 4.5 ± 0.6 (168 h p.i.) for (111)In-D2B IgG, (111)In-F(ab')2, (111)In-Fab and (111)In-capromab pendetide, respectively. LNCaP tumors were clearly visualized with microSPECT with all antibody formats. This study demonstrates the feasibility of D2B IgG, F(ab')2 and Fab fragments for targeting PSMA-expressing prostate cancer xenografts. Copyright © 2014 John Wiley & Sons, Ltd.

  9. Morphometric analysis of instar variation in Tenebrio molitor (Coleoptera: Tenebrionidae)

    Science.gov (United States)

    Measurements of head capsule, mandible, metanotum, and body weight were done on larvae of Tenebrio molitor L. (Coleoptera: Tenebrionide) from the second to the last instar. Instar number varied from 14 to 18, but 15 or 16 instars were the most common. The value of dimensional measurements was evalua...

  10. Attractant and disruptant semiochemicals for Dendroctonus jeffreyi (Coleoptera: Curculionidae: Scolytinae)

    Science.gov (United States)

    Brian Strom; Smith S.L.; Brownie C.

    2013-01-01

    Jeffrey pine, Pinus jeffreyi Greville and Balfour, is a dominant yellow pine and important overstory component of forests growing on diverse sites from southwestern Oregon to Baja California to western Nevada. The Jeffrey pine beetle, Dedroctonus jeffreyi Hopkins (Coleoptera: Curculionidae: Scolytinae), is monophagous on Jeffrey...

  11. A coffee berry borer (Coleoptera: Curculionidae: Scolytinae) bibliography

    Science.gov (United States)

    One hundred years ago, one of the most significant biological invasions of an agricultural insect pest in the Americas was initiated. Endemic to Africa, the coffee berry borer (Hypothenemus hampei; Coleoptera: Curculionidae) was accidentally introduced to Brazil in 1913 and years later invaded coffe...

  12. Changing distributions of Cantharidae and Buprestidae within Great Britain (Coleoptera)

    NARCIS (Netherlands)

    Alexander, K.

    2003-01-01

    Changing distributions of Cantharidae and Buprestidae within Great Britain (Coleoptera) Data are presented on the distribution of selected species from two coleopteran families chosen to represent a random slice of the British fauna. The species have been chosen as exhibiting extremes of range

  13. Using Malaise traps to sample ground beetles (Coleoptera: Carabidae)

    Science.gov (United States)

    Michael D. Ulyshen; James L. Hanula; Scott Horn

    2005-01-01

    Pitfall traps provide an easy and inexpensive way to sample ground-dwelling arthropods (Spence and Niemela 1994; Spence et al. 1997; Abildsnes and Tommeras 2000) and have been used exclusively in many studies of the abundance and diversity of ground beetles (Coleoptera: Carabidae). Despite the popularity of this trapping technique, pitfall traps have many disadvantages...

  14. The Peritelini (Coleoptera: Curculionidae, Entiminae of the Vitale collection

    Directory of Open Access Journals (Sweden)

    Cosimo Baviera

    2016-01-01

    Full Text Available The collection of Peritelini (Coleoptera Curculionidae Entiminae currently stored in the Vitale collection of Messina University is an element of great importance for studies of taxonomy and biogeography of these rarely collected weevils. All species are commented in relation to the contributions to this taxonomic group, published on several occasions by the authors.

  15. LC-MS/MS quantification of free and Fab-bound colchicine in plasma, urine and organs following colchicine administration and colchicine-specific Fab fragments treatment in Göttingen minipigs.

    Science.gov (United States)

    Fabresse, Nicolas; Allard, Julien; Sardaby, Marine; Thompson, Adrian; Clutton, R Eddie; Eddleston, Michael; Alvarez, Jean-Claude

    2017-08-15

    Clinical evaluation of a colchicine specific antigen-binding fragment (Fab) in order to treat colchicine poisoning required the development of an accurate method allowing quantification of free and Fab-bound colchicine in plasma and urine, and free colchicine in tissues, to measure colchicine redistribution after Fab administration. Three methods have been developed for this purpose, and validated in plasma, urine and liver: total colchicine was determined after denaturation of Fab by dilution in water and heating; free colchicine was separated from Fab-bound colchicine by filtration with 30KDa micro-filters; tissues were homogenized in a tissue mixer. Deuterated colchicine was used as internal standard. Samples were extracted by liquid-liquid extraction and analyzed with a LC-MS/MS. LOQ were 0.5ng/mL in plasma and urine for free and total colchicine and 5pg/mg in tissues. The methods were linear in the 0.5-100ng/mL range in plasma and urine, and 5-300pg/mg in tissues with determination coefficients>0.99. Precision and accuracy of QC samples presented a CVFab fragments. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures

    Science.gov (United States)

    2013-01-01

    Background Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Results Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Conclusions Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into

  17. Double mutation of cell wall proteins CspB and PBP1a increases secretion of the antibody Fab fragment from Corynebacterium glutamicum

    Science.gov (United States)

    2014-01-01

    Background Among other advantages, recombinant antibody-binding fragments (Fabs) hold great clinical and commercial potential, owing to their efficient tissue penetration compared to that of full-length IgGs. Although production of recombinant Fab using microbial expression systems has been reported, yields of active Fab have not been satisfactory. We recently developed the Corynebacterium glutamicum protein expression system (CORYNEX®) and demonstrated improved yield and purity for some applications, although the system has not been applied to Fab production. Results The Fab fragment of human anti-HER2 was successfully secreted by the CORYNEX® system using the conventional C. glutamicum strain YDK010, but the productivity was very low. To improve the secretion efficiency, we investigated the effects of deleting cell wall-related genes. Fab secretion was increased 5.2 times by deletion of pbp1a, encoding one of the penicillin-binding proteins (PBP1a), mediating cell wall peptidoglycan (PG) synthesis. However, this Δpbp1a mutation did not improve Fab secretion in the wild-type ATCC13869 strain. Because YDK010 carries a mutation in the cspB gene encoding a surface (S)-layer protein, we evaluated the effect of ΔcspB mutation on Fab secretion from ATCC13869. The Δpbp1a mutation showed a positive effect on Fab secretion only in combination with the ΔcspB mutation. The ΔcspBΔpbp1a double mutant showed much greater sensitivity to lysozyme than either single mutant or the wild-type strain, suggesting that these mutations reduced cell wall resistance to protein secretion. Conclusion There are at least two crucial permeability barriers to Fab secretion in the cell surface structure of C. glutamicum, the PG layer, and the S-layer. The ΔcspBΔpbp1a double mutant allows efficient Fab production using the CORYNEX® system. PMID:24731213

  18. Trastuzumab- and Fab' fragment-modified curcumin PEG-PLGA nanoparticles: preparation and evaluation in vitro and in vivo.

    Science.gov (United States)

    Duan, Dongyu; Wang, Aiping; Ni, Ling; Zhang, Liping; Yan, Xiuju; Jiang, Ying; Mu, Hongjie; Wu, Zimei; Sun, Kaoxiang; Li, Youxin

    2018-01-01

    Nanoparticles (NPs) modified with bio-ligands represent a promising strategy for active targeted drug delivery to tumour. However, many targeted ligands, such as trastuzumab (TMAB), have high molecular weight, limiting their application for targeting. In this study, we prepared Fab' (antigen-binding fragments cut from TMAB)-modified NPs (Fab'-NPs) with curcumin (Cur) as a model drug for more effective targeting of human epidermal growth factor receptor 2 (HER2/ErbB2/Neu), which is overexpressed on breast cancer cells. The release kinetics was conducted by dialysis bags. The ability to kill HER2-overexpressing BT-474 cells of Fab'-Cur-NPs compared with TMAB-Cur-NPs was conducted by cytotoxicity experiments. Qualitative and quantitative cell uptake studies using coumarin-6 (fluorescent probe)-loaded NPs were performed by fluorescence microscopy and flow cytometry. Pharmacokinetics and biodistribution experiments in vivo were assessed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The release kinetics showed that both Fab'-Cur-NPs and TMAB-Cur-NPs provided continuous, slow release of curcumin for 72 h, with no significant difference. In vitro cytotoxicity experiments showed that Fab'-Cur-NPs manifested prominent ability to kill HER2-overexpressing BT-474 cells compared with TMAB-Cur-NPs. Qualitative and quantitative cell uptake studies indicated that the accumulation of Fab'-NPs was greater than that of TMAB-NPs in BT-474 (HER2+) cells; However, there was no significant difference in MDA-MB-231 (HER2-) cells. Pharmacokinetics and biodistribution experiments in vivo demonstrated that the half-life (t1/2) and area under the blood concentration-time curve (AUC0-t) of Fab'-Cur-NPs increased 5.30-fold and 1.76-fold relative to those of TMAB-Cur-NPs, respectively. Furthermore, the tumor accumulation of Fab'-Cur-NPs was higher than that of TMAB-Cur-NPs. Fab' fragment has greater capacity than the intact antibody to achieve tumor targeting through NP

  19. Predation by Flat Bark Beetles (Coleoptera: Silvanidae and Laemophloeidae) on Coffee Berry Borer (Coleoptera: Curculionidae) in Hawaii coffee

    Science.gov (United States)

    Coffee berry borer(CBB), Hypothenemus hampei, is a serious pest of coffee worldwide and a new invasive pest in Hawaii. Adult flat bark beetles, mainly Leptophloeus sp.(75%) and Cathartus quadricollis(21%) (Coleoptera: Laemophloeidae and Silvanidae, respectively), were found feeding in CBB-infested c...

  20. Tumour imaging by the detection of fibrin clots in tumour stroma using an anti-fibrin Fab fragment

    Science.gov (United States)

    Obonai, Toshifumi; Fuchigami, Hirobumi; Furuya, Fumiaki; Kozuka, Naoyuki; Yasunaga, Masahiro; Matsumura, Yasuhiro

    2016-01-01

    The diagnosis of early and aggressive types of cancer is important for providing effective cancer therapy. Cancer-induced fibrin clots exist only within lesions. Previously, we developed a monoclonal antibody (clone 102-10) that recognizes insoluble fibrin but not fibrinogen or soluble fibrin and confirmed that fibrin clots form continuously in various cancers. Here, we describe the development of a Fab fragment probe of clone 102-10 for tumour imaging. The distribution of 102-10 Fab was investigated in genetically engineered mice bearing pancreatic ductal adenocarcinoma (PDAC), and its effect on blood coagulation was examined. Immunohistochemical and ex vivo imaging revealed that 102-10 Fab was distributed selectively in fibrin clots in PDAC tumours 3 h after injection and that it disappeared from the body after 24 h. 102-10 Fab had no influence on blood coagulation or fibrinolysis. Tumour imaging using anti-fibrin Fab may provide a safe and effective method for the diagnosis of invasive cancers by detecting fibrin clots in tumour stroma. PMID:27009516

  1. Feasibility study of the Fab fragment of a monoclonal antibody against tissue factor as a diagnostic tool.

    Science.gov (United States)

    Tsumura, Ryo; Sato, Ryuta; Furuya, Fumiaki; Koga, Yoshikatsu; Yamamoto, Yoshiyuki; Fujiwara, Yuki; Yasunaga, Masahiro; Matsumura, Yasuhiro

    2015-12-01

    Tissue factor (TF) is expressed strongly in various types of cancer, especially cancers that are often refractory to treatment, such as pancreatic cancer. In this study, we compared the differences in the biophysical and pharmacological properties of whole IgG and the Fab fragment of anti-human TF monoclonal antibody (1849 antibodies), in order to determine their suitability for application in the diagnosis and treatment of cancers. In the biophysical examination, we investigated the characteristics of 1849-whole IgG and 1849-Fab by SPR sensing and confocal fluorescence microscopy analysis using recombinant human TF antigen and TF-overexpressing human pancreatic cancer cell line, BxPC3, respectively. After conjugation with Alexa-Flour-647, in vivo imaging was conducted in mice bearing BxPC3 xenograft tumors. Furthermore, the distribution of the conjugates in tumors and major organs was evaluated by ex vivo study. The in vitro experiments showed that 1849 antibodies had high affinity against TF antigen. In addition, 1849-Fab showed a faster dissociation rate from the antigen than 1849-whole IgG. In mice, 1849-Fab-Alexa-Flour-647 showed rapid renal clearance and faster tumor accumulation, achieving a high contrast signal over nearby normal tissues in the early phase and enhanced tumor penetration after administration. On the other hand, 1849-whole IgG-Alexa-Flour-647 showed slow clearance from the blood and sustained high tumor accumulation. These results suggest that 1849-Fab may be a useful tool for pancreatic cancer diagnosis.

  2. The Fatty Acid Biosynthesis Enzyme FabI Plays a Key Role In the Development of Liver Stage Malarial Parasites

    Science.gov (United States)

    Yu, Min; Santha Kumar, T. R.; Nkrumah, Louis J.; Coppi, Alida; Retzlaff, Silke; Li, Celeste D.; Kelly, Brendan J.; Moura, Pedro A.; Lakshmanan, Viswanathan; Freundlich, Joel S.; Valderramos, Juan-Carlos; Vilcheze, Catherine; Siedner, Mark; Tsai, Jennifer H.-C.; Falkard, Brie; Sidhu, Amar bir Singh; Purcell, Lisa A.; Gratraud, Paul; Kremer, Laurent; Waters, Andy P.; Schiehser, Guy; Jacobus, David P.; Janse, Chris J.; Ager, Arba; Jacobs, William R.; Sacchettini, James C.; Heussler, Volker; Sinnis, Photini; Fidock, David A.

    2008-01-01

    SUMMARY Fatty acid biosynthesis has been viewed as an important biological function of and therapeutic target for Plasmodium falciparum asexual blood stage infection. This apicoplast-resident type II pathway, distinct from the mammalian type I process, includes FabI. Here, we report synthetic chemistry and transfection studies concluding that Plasmodium FabI is not the target of the antimalarial activity of the bacterial FabI inhibitor triclosan. Disruption of fabI in P. falciparum or the rodent parasite P. berghei does not impede blood stage growth. In contrast, mosquito-derived fabI-deficient P. berghei sporozoites are markedly less infective for mice and typically fail to complete liver stage development in vitro. This is characterized by an inability to form intra-hepatic merosomes that normally initiate blood stage infections. These data illuminate key differences between liver and blood stage parasites in their requirements for host versus de novo synthesized fatty acids, and create new prospects for stage-specific antimalarial interventions. PMID:19064257

  3. Metazoan parasites of Mandi-amarelo Pimelodus maculatus and of Jundiá Rhamdia quelen (Osteichthyes: Siluriformes) of Paraíba do Sul River, Volta Redonda, Rio de Janeiro.

    Science.gov (United States)

    Venancio, Aline Cristine Pinto; de Aguiar, Gesilene Ribeiro; Lopes, Patrícia da Silva; Alves, Dimitri Ramos

    2010-01-01

    Forty-one specimens of mandi-amarelo Pimelodus maculatus Lacépède, 1803 (Siluriformes: Pimelodidae) and 54 specimens of jundiá Rhamdia quelen (Quoy & Gaimard, 1824) (Siluriformes: Heptapteridae) were collected from the Paraíba do Sul River, Volta Redonda, State of Rio de Janeiro, Brazil between November 2007 and October 2008. These fish underwent necropsy so their infracommunities of metazoan parasites could be studied. The same three species of parasites were collected in the two fish species studied. These were one monogenean, one nematode, and one hirudinean. Cucullanus pinnai (Travassos, Artiga, and Pereira, 1928) (Nematoda: Cucullanidae) and Aphanoblastella sp. (Monogenea: Dactylogyridae) were the dominant species with the highest prevalence in P. maculatus and R. quelen. The parasite species of P. maculatus and R. quelen showed an atypical over-dispersed pattern of distribution. No parasite species showed significant correlation between the body total length of the siluriform hosts and their prevalence and abundance. The parasite species richness showed a mean value of 0.87 ± 0.67 (0-2) and 0.57 ± 0.56 (0-2) in P. maculatus and R. quelen, respectively, and no correlation with the body total length.

  4. [The endoparasitic helminths of Pimelodus maculatus Lacépède, 1803 Siluriformes, Pimelodidae) from the two localities (Lagoon and gutter of the River) of the Guandu River, State of Rio de Janeiro, Brazil].

    Science.gov (United States)

    Albuquerque, Marcia C; Santos, Michelle D; Monteiro, Cassandra M; Martins, Amanda N; Ederli, Nicole B; Brasil-Sato, Marilia C

    2008-09-01

    Between November 2003 and March 2004, fourty specimens of Pimelodus maculatus Lacépède, 1803 from Guandu River and thirty-nine from Guandu Lagoon (Nova Iguaçu, RJ) were collected, for the analysis of endoparasitic fauna. A total of 236 specimens of Cucullanus pinnai Travassos, Artigas & Pereira, 1928 (Nematoda, Cucullanidae) were collected, being 163 adult specimens in the gut, three in the stomach and 70 larvae in the celomatic cavity and seven specimens of adults Nomimoscolex sp. (Eucestoda, Proteocephalidea) in the gut were found. Cucullanus pinnai presented prevalence (P) 77.50%, mean intensity (MI) 3.40 and mean abundance (MA) 2.60 on River and P: 66.67%, MI: 5.04, MA: 3.36 on Lagoon. Nomimoscolex sp. presented on River P: 2.50%, MI: 2.00, MA: 0.05, and P: 10.26%, MI: 1.25, MA: 0.13 on Lagoon. There was not significant positive interspecific association on the lagoon. In this research, the endoparasitic richness of P. maculatus was scarcest than similar studies in Guandu River and others rivers of different basins. The results about C. pinnai could be suggesting that the cycle of C. pinnai evolve only a host, occurring a histotrophic fase, in this case, in P. maculatus. Periodic analysis of the endoparasites indices in P. maculatus through the years may be used to describe the hydric quality of the Guandu River.

  5. Activity of the thyroid and the pituitary gland in the viviparous Cyprinodonts Lebistes reticulatus (Peters), Xiphophorus helleri Heckel and Xiphophorus maculatus Günther during the development of the gonopodium

    NARCIS (Netherlands)

    Stolk, A.

    1959-01-01

    A description is given of the activity of the thyroid and of the pituitary gland during the development of the gonopodium in the viviparous Cyprinodonts Lebistes reticulatus (Peters), Xiphophorus helleri (Heckel) and Xiphophorus maculatus (Günther). During this process the thyroid gland and the

  6. Feeding ω-3 PUFA enriched rotifers to Galaxias maculatus (Jenyns, 1842 larvae reared at different salinity conditions: effects on growth parameters, survival and fatty acids profile

    Directory of Open Access Journals (Sweden)

    Patricio Dantagnan

    2013-07-01

    Full Text Available Despite the well known importance of ω-3 polyunsaturated fatty acids (PUFA in marine and freshwater fish larvae, there are few studies on how essential fatty acid requirements and composition on whole body can be altered by changes in water salinity. The present study aimed to determine the effect of salinity on ω-3 PUFA requirements, larval growth survival and fatty acid composition of Galaxias maculatus larvae cultured at two different salinities (0 and 15 g L-1 for 20 days while fed rotifers containing two different levels of ω-3 PUFA (1.87 and 3.16%. The results denoted a marked difference in ω-3 PUFA requirements and in the pattern of fatty acid deposition in the whole body of larvae reared at different salinities, depending of ω-3 PUFA in diets. Thus, to improve growth and survival larvae of G. maculatus reared at 0 g L-1 require higher levels of ω-3 PUFA, principally 18:3 ω-3. Larvae reared at salinities of 15 g L-1 require low levels of ω-3 PUFA for optimal survival, especially 18:3 ω-3. Eicosapentaenoic acid and docosahexaenoic acid content in the whole body of larvae was also affected by water salinity.

  7. Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins) with high toxicity to the larvae of the bruchid Callosobruchus maculatus

    Energy Technology Data Exchange (ETDEWEB)

    Souza, A.J. [Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ (Brazil); Ferreira, A.T.S.; Perales, J.; Beghini, D.G. [Laboratório de Toxinologia, Departamento de Fisiologia e Farmacodinâmica, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ (Brazil); Fernandes, K.V.S.; Xavier-Filho, J.; Venancio, T.M.; Oliveira, A.E.A. [Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ (Brazil)

    2011-01-27

    Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitinbinding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.

  8. Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins) with high toxicity to the larvae of the bruchid Callosobruchus maculatus

    International Nuclear Information System (INIS)

    Souza, A.J.; Ferreira, A.T.S.; Perales, J.; Beghini, D.G.; Fernandes, K.V.S.; Xavier-Filho, J.; Venancio, T.M.; Oliveira, A.E.A.

    2011-01-01

    Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitinbinding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation

  9. Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins) with high toxicity to the larvae of the bruchid Callosobruchus maculatus.

    Science.gov (United States)

    Souza, A J; Ferreira, A T S; Perales, J; Beghini, D G; Fernandes, K V S; Xavier-Filho, J; Venancio, T M; Oliveira, A E A

    2012-02-01

    Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitin-binding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.

  10. Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins with high toxicity to the larvae of the bruchid Callosobruchus maculatus

    Directory of Open Access Journals (Sweden)

    A.J. Souza

    2012-02-01

    Full Text Available Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitin-binding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.

  11. Spinal cord self-repair during tail regeneration in Polypedates maculatus and putative role of FGF1 as a neurotrophic factor.

    Science.gov (United States)

    Hota, Jutshina; Pati, Sushri Sangita; Mahapatra, Pravati Kumari

    2018-03-01

    Spinal cord injury could be fatal in man and often results in irreversible medical conditions affecting mobility. However, anuran amphibians win over such pathological condition by the virtue of regeneration abilities. The tail of anuran tadpoles therefore allures researchers to study spinal cord injury and self- repair process. In the present study, we inflicted injury to the spinal cord by means of surgical transection of the tail and investigated the self-repair activity in the tadpoles of the Indian tree frog Polypedates maculatus. We also demonstrate for the first time by immunofluorescence localization the expression pattern of Fibroblast Growth Factor1 (FGF1) during spinal cord regeneration which has not been documented earlier in anurans. FGF1, bearer of the mitogenic and neurotrophic properties seems to be expressed by progenitor cells that facilitate regeneration. Spinal cord during tail regeneration in P. maculatus attains functional recovery within a span of 2 weeks thus enabling the organism to survive in an aquatic medium till metamorphosis. Moreover, during the course of spinal cord regeneration in the regenerating tail, melanocytes showed an interesting behaviour as these neural crest derivatives were missing near the early regenerates until their reappearance where they were positioned in close proximity with the regenerated spinal cord as in the normal tail. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Improving Fab' fragment retention in an autonucleolytic Escherichia coli strain by swapping periplasmic nuclease translocation signal from OmpA to DsbA.

    Science.gov (United States)

    Schofield, Desmond M; Sirka, Ernestas; Keshavarz-Moore, Eli; Ward, John M; Nesbeth, Darren N

    2017-12-01

    To reduce unwanted Fab' leakage from an autonucleolytic Escherichia coli strain, which co-expresses OmpA-signalled Staphylococcal nuclease and Fab' fragment in the periplasm, by substituting in Serratial nuclease and the DsbA periplasm translocation signal as alternatives. We attempted to genetically fuse a nuclease from Serratia marcescens to the OmpA signal peptide but plasmid construction failed, possibly due to toxicity of the resultant nuclease. Combining Serratial nuclease to the DsbA signal peptide was successful. The strain co-expressing this nuclease and periplasmic Fab' grew in complex media and exhibited nuclease activity detectable by DNAse agar plate but its growth in defined medium was retarded. Fab' coexpression with Staphylococcal nuclease fused to the DsbA signal peptide resulted in cells exhibiting nuclease activity and growth in defined medium. In cultivation to high cell density in a 5 l bioreactor, DsbA-fused Staphylococcal nuclease co-expression coincided with reduced Fab' leakage relative to the original autonucleolytic Fab' strain with OmpA-fused staphylococcal nuclease. We successfully rescued Fab' leakage back to acceptable levels and established a basis for future investigation of the linkage between periplasmic nuclease expression and leakage of co-expressed periplasmic Fab' fragment to the surrounding growth media.

  13. The strength of small: Improved targeting of Insulin-like Growth Factor-1 Receptor (IGF-1R) with F(ab')2-R1507 fragments in Ewing sarcomas

    NARCIS (Netherlands)

    Fleuren, Emmy D. G.; Versleijen-Jonkers, Yvonne M. H.; Heskamp, Sandra; Roeffen, Melissa H. S.; Bouwman, Wilbert H.; Molkenboer-Kuenen, Janneke D. M.; van Laarhoven, Hanneke W. M.; Oyen, Wim J. G.; Boerman, Otto C.; van der Graaf, Winette T. A.

    2013-01-01

    To investigate whether F(ab')2-fragments of the monoclonal Insulin-like Growth Factor-1 Receptor (IGF-1R) antibody R1507 (F(ab')2-R1507) can successfully target IGF-1R in Ewing sarcomas (ES). BALB/c nude mice were subcutaneously implanted with IGF-1R-expressing human ES xenografts (EW-5 and EW-8)

  14. COPROPHANAEUS MORENOI ARNAUD, 1982 (COLEOPTERA: SCARABAEIDAE: SCARABAEINAE) IN THE GORGONA NATIONAL NATURAL PARK (COLOMBIAN PACIFIC OCEAN)

    OpenAIRE

    Cultid Medina, Carlos A; Giraldo López, Alan

    2012-01-01

    A new record of the dung beetle Coprophanaeus morenoi Arnaud, 1982 (Coleoptera: Scarabaeidae: Scarabaeinae), for the Gorgona Natural National Park (Gorgona NNP, Colombia) in the Colombian Pacific Ocean is presented. Se presenta un nuevo registro del escarabajo estercolero Coprophanaeus morenoi Arnaud, 1982 (Coleoptera: Scarabaeidae: Scarabaeinae), para el Parque Natural Nacional Gorgona, en el océano Pacifico colombiano.

  15. Het voorkomen van de glanskevers van het genus Caprophilus in Nederland (Coleoptera: Nitidulidae)

    NARCIS (Netherlands)

    Oude, de J.

    2005-01-01

    The occurrence of nitidulid beetles of the genus Carpophilus in the Netherlands (Coleoptera: Nitidulidae) Some species of the genus Carpophilus (Coleoptera: Nitidulidae) may act as pests of stored products like cocoabeans, peanuts, dried fruits like figs and dates, imported from tropical or

  16. Expression, purification and characterization of enoyl-ACP reductase II, FabK, from Porphyromonas gingivalis

    Energy Technology Data Exchange (ETDEWEB)

    Hevener, Kirk E.; Mehboob, Shahila; Boci, Teuta; Truong, Kent; Santarsiero, Bernard D.; Johnson, Michael E. (UIC)

    2012-10-25

    The rapid rise in bacterial drug resistance coupled with the low number of novel antimicrobial compounds in the discovery pipeline has led to a critical situation requiring the expedient discovery and characterization of new antimicrobial drug targets. Enzymes in the bacterial fatty acid synthesis pathway, FAS-II, are distinct from their mammalian counterparts, FAS-I, in terms of both structure and mechanism. As such, they represent attractive targets for the design of novel antimicrobial compounds. Enoyl-acyl carrier protein reductase II, FabK, is a key, rate-limiting enzyme in the FAS-II pathway for several bacterial pathogens. The organism, Porphyromonas gingivalis, is a causative agent of chronic periodontitis that affects up to 25% of the US population and incurs a high national burden in terms of cost of treatment. P. gingivalis expresses FabK as the sole enoyl reductase enzyme in its FAS-II cycle, which makes this a particularly appealing target with potential for selective antimicrobial therapy. Herein we report the molecular cloning, expression, purification and characterization of the FabK enzyme from P. gingivalis, only the second organism from which this enzyme has been isolated. Characterization studies have shown that the enzyme is a flavoprotein, the reaction dependent upon FMN and NADPH and proceeding via a Ping-Pong Bi-Bi mechanism to reduce the enoyl substrate. A sensitive assay measuring the fluorescence decrease of NADPH as it is converted to NADP{sup +} during the reaction has been optimized for high-throughput screening. Finally, protein crystallization conditions have been identified which led to protein crystals that diffract x-rays to high resolution.

  17. Effect of acetylation on monoclonal antibody ZCE-025 Fab': Distribution in normal and tumor-bearing mice

    International Nuclear Information System (INIS)

    Tarburton, J.P.; Halpern, S.E.; Hagan, P.L.; Sudora, E.; Chen, A.; Fridman, D.M.; Pfaff, A.E.

    1990-01-01

    Studies were performed to determine in vitro and in vivo effects of acetylation on Fab' fragments of ZCE-025, a monoclonal anti-CEA antibody. Isoelectric focusing revealed a drop in isoelectric point of 1.7 pI units following acetylation. Biodistribution studies of acetylated and nonacetylated [111In]Fab' were performed in normal BALB/c mice and in nude mice bearing the T-380 CEA-producing human colon tumor. The acetylated fragments remained in the vascular compartment longer and had significantly diminished renal uptake of 111In compared to controls. While acetylation itself effected a 50% drop in immunoreactivity, tumor uptake of the acetylated and nonacetylated 111In-labeled Fab' fragments was comparable, with the exception of one data point, through 72 h

  18. Successful use of digoxin-specific immune Fab in the treatment of severe Nerium oleander toxicosis in a dog.

    Science.gov (United States)

    Pao-Franco, Amaris; Hammond, Tara N; Weatherton, Linda K; DeClementi, Camille; Forney, Scott D

    2017-09-01

    To describe a case in which digoxin-specific immune Fab was used successfully in a dog with severe oleander toxicosis secondary to ingesting plant material. A 6-year-old intact female Rhodesian Ridgeback mixed breed dog was presented for severe oleander toxicosis and was refractory to all antiarrhythmic therapies and supportive care. Digoxin-specific immune Fab was successful in treating this dog. The dog recovered but suffered ischemic injuries, the long-term effects of which are unknown. This report describes the successful use of digoxin-specific immune Fab in the treatment of oleander toxicosis in a dog, which has not previously been published in veterinary literature. Oleander poisoning can be associated with permanent cardiac arrhythmias due to the ischemic damage. © Veterinary Emergency and Critical Care Society 2017.

  19. Dosimetry of 99mTc-DD-3B6/22 Fab' for use in staging ovarian cancer

    International Nuclear Information System (INIS)

    Hetherington, E.; Smith, S.V.; Schmidt, P.F.; Di Bartolo, N.M.; Prabakaran, K.; Femandes, V.; Donaghy, T.; Clingan, P.; Bundesen, P.; Hillyard, C.

    1998-01-01

    Full text: The diagnostic utility of 99 mTc-DD-3B6/22 Fab'' for staging ovarian cancer in a phase 11 trial is under way. The dose due to 99 mTc- DD-3B6/22 Fab'' was used to estimate the dose for the analogous 188 Re compound for therapy of ovarian cancer. The new agent, 99 mTc-DD-3B6/22 Fab'' (600 mBq) was found to clear rapidly from the blood via the renal system. A ''kidney'' phantom was used to calibrate renal activity. The dose to selected organs was estimated using the MlRDose 2 adult female model. In most cases the kidney uptake accounted for 30% of activity administered. Urine activity was measured directly and the dose calculated assuming a mean volume of 300 mL. All remaining activity was assumed to be uniformly distributed throughout the body. Results show the dose to kidneys, bladder wall and whole body were 4.5-9.1; 1.2-6.3; 0.2-0.3 cGy, respectively. The biological distribution of the 188 Re-DD-3B6/22 Fab'' was assumed to be similar to that of 99 mTc-DD-3B6/22 Fab''. The activity/time data was adjusted for the relative t 1/2 of 99 mTc and 188 Re. The calculated kidney dose due to 188 Re was approx. 800 cGy. As kidney damage from acute radiation nephritis is thought to occur at doses >1000 cGy, this study indicates that provided uptake at tumor site is adequate 188 Re-DD-3B6/22 Fab'' may have a role in therapy of ovarian cancer

  20. A study on EUV reticle surface molecular contamination under different storage conditions in a HVM foundry fab

    Science.gov (United States)

    Singh, SherJang; Yatzor, Brett; Taylor, Ron; Wood, Obert; Mangat, Pawitter

    2017-03-01

    The prospect of EUVL (Extreme Ultraviolet Lithography) insertion into HVM (High Volume Manufacturing) has never been this promising. As technology is prepared for "lab to fab" transition, it becomes important to comprehend challenges associated with integrating EUVL infrastructure within existing high volume chip fabrication processes in a foundry fab. The existing 193nm optical lithography process flow for reticle handling and storage in a fab atmosphere is well established and in-fab reticle contamination concerns are mitigated with the reticle pellicle. However EUVL reticle pellicle is still under development and if available, may only provide protection against particles but not molecular contamination. HVM fab atmosphere is known to be contaminated with trace amounts of AMC's (Atmospheric Molecular Contamination). If such contaminants are organic in nature and get absorbed on the reticle surface, EUV photon cause photo-dissociation resulting into carbon generation which is known to reduce multilayer reflectivity and also degrades exposure uniformity. Chemical diffusion and aggregation of other ions is also reported under the e-beam exposure of a EUV reticle which is known to cause haze issues in optical lithography. Therefore it becomes paramount to mitigate absorbed molecular contaminant concerns on EUVL reticle surface. In this paper, we have studied types of molecular contaminants that are absorbed on an EUVL reticle surface under HVM fab storage and handling conditions. Effect of storage conditions (gas purged vs atmospheric) in different storage pods (Dual pods, Reticle Clamshells) is evaluated. Absorption analysis is done both on ruthenium capping layer as well as TaBN absorber. Ru surface chemistry change as a result of storage is also studied. The efficacy of different reticle cleaning processes to remove absorbed contaminant is evaluated as well.

  1. Structure of FabH and factors affecting the distribution of branched fatty acids in Micrococcus luteus.

    Science.gov (United States)

    Pereira, Jose H; Goh, Ee-Been; Keasling, Jay D; Beller, Harry R; Adams, Paul D

    2012-10-01

    Micrococcus luteus is a Gram-positive bacterium that produces iso- and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters [coenzyme A (CoA) or acyl carrier protein (ACP)]; this activity is of interest for the production of advanced biofuels. In an effort to better understand the control of the formation of branched fatty acids in M. luteus, the structure of FabH (MlFabH) was determined. FabH, or β-ketoacyl-ACP synthase III, catalyzes the initial step of fatty-acid biosynthesis: the condensation of malonyl-ACP with an acyl-CoA. Analysis of the MlFabH structure provides insights into its substrate selectivity with regard to length and branching of the acyl-CoA. The most structurally divergent region of FabH is the L9 loop region located at the dimer interface, which is involved in the formation of the acyl-binding channel and thus limits the substrate-channel size. The residue Phe336, which is positioned near the catalytic triad, appears to play a major role in branched-substrate selectivity. In addition to structural studies of MlFabH, transcriptional studies of M. luteus were also performed, focusing on the increase in the ratio of anteiso:iso-branched alkenes that was observed during the transition from early to late stationary phase. Gene-expression microarray analysis identified two genes involved in leucine and isoleucine metabolism that may explain this transition.

  2. Structure of FabH and factors affecting the distribution of branched fatty acids in Micrococcus luteus

    International Nuclear Information System (INIS)

    Pereira, Jose H.; Goh, Ee-Been; Keasling, Jay D.; Beller, Harry R.; Adams, Paul D.

    2012-01-01

    In an effort to better understand the control of the formation of branched fatty acids in Micrococcus luteus, the structure of β-ketoacyl-ACP synthase III, which catalyzes the initial step of fatty-acid biosynthesis, has been determined. Micrococcus luteus is a Gram-positive bacterium that produces iso- and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters [coenzyme A (CoA) or acyl carrier protein (ACP)]; this activity is of interest for the production of advanced biofuels. In an effort to better understand the control of the formation of branched fatty acids in M. luteus, the structure of FabH (MlFabH) was determined. FabH, or β-ketoacyl-ACP synthase III, catalyzes the initial step of fatty-acid biosynthesis: the condensation of malonyl-ACP with an acyl-CoA. Analysis of the MlFabH structure provides insights into its substrate selectivity with regard to length and branching of the acyl-CoA. The most structurally divergent region of FabH is the L9 loop region located at the dimer interface, which is involved in the formation of the acyl-binding channel and thus limits the substrate-channel size. The residue Phe336, which is positioned near the catalytic triad, appears to play a major role in branched-substrate selectivity. In addition to structural studies of MlFabH, transcriptional studies of M. luteus were also performed, focusing on the increase in the ratio of anteiso:iso-branched alkenes that was observed during the transition from early to late stationary phase. Gene-expression microarray analysis identified two genes involved in leucine and isoleucine metabolism that may explain this transition

  3. Radioimmunoimaging of human colon carcinoma grafted into nudemice using 131I-labeled monoclonal anticea antibody and its F(ab')2 fragments

    International Nuclear Information System (INIS)

    Liu Guangda

    1988-01-01

    131 I-labeled monoclonal anti-CEA antibody and its F(ab') 2 fragments were injected into nude mice bearing human colon carcinoma xenografts for tumor localization and radioimmunoimaging studies. Transplanted tumors were visualized in 12 hours after injection of the labeled anti-CEA or its F(ab') 2 by gamma camera. Biodistribution data indicated that F(ab') 2 fragments were cleared more rapidly from blood (T 1/2 = 13.3 h for F(ab') 2 , T 1/2 = 21.1 h for intact antibody) over 6-24 h and had higher tumor blood ratios. The intact antibody was concentrated in the tumor better than F(ab') 2 . In double-label experiments, a nonspecific localization of the control ( 125 I-labeled anti-HCG) in the tumor was also observed

  4. ImmunoPET of tissue factor expression in triple-negative breast cancer with a radiolabeled antibody Fab fragment

    International Nuclear Information System (INIS)

    Shi, Sixiang; Hong, Hao; Orbay, Hakan; Yang, Yunan; Ohman, Jakob D.; Graves, Stephen A.; Nickles, Robert J.; Liu, Bai; Wong, Hing C.; Cai, Weibo

    2015-01-01

    To date, there is no effective therapy for triple-negative breast cancer (TNBC), which has a dismal clinical outcome. Upregulation of tissue factor (TF) expression leads to increased patient morbidity and mortality in many solid tumor types, including TNBC. Our goal was to employ the Fab fragment of ALT-836, a chimeric anti-human TF mAb, for PET imaging of TNBC, which can be used to guide future TNBC therapy. ALT-836-Fab was generated by enzymatic papain digestion. SDS-PAGE and FACS studies were performed to evaluate the integrity and TF binding affinity of ALT-836-Fab before NOTA conjugation and 64 Cu-labeling. Serial PET imaging and biodistribution studies were carried out to evaluate the tumor targeting efficacy and pharmacokinetics in the MDA-MB-231 TNBC model, which expresses high levels of TF on the tumor cells. Blocking studies, histological assessment, as well as RT-PCR were performed to confirm TF specificity of 64 Cu-NOTA-ALT-836-Fab. ALT-836-Fab was produced with high purity, which exhibited superb TF binding affinity and specificity. Serial PET imaging revealed rapid and persistent tumor uptake of 64 Cu-NOTA-ALT-836-Fab (5.1 ± 0.5 %ID/g at 24 h post-injection; n = 4) and high tumor/muscle ratio (7.0 ± 1.2 at 24 h post-injection; n = 4), several-fold higher than that of the blocking group and tumor models that do not express significant level of TF, which was confirmed by biodistribution studies. TF specificity of the tracer was also validated by histology and RT-PCR. 64 Cu-NOTA-ALT-836-Fab exhibited prominent tissue factor targeting efficiency in MDA-MB-231 TNBC model. The use of a Fab fragment led to fast tumor uptake and good tissue/muscle ratio, which may be translated into same-day immunoPET imaging in the clinical setting to improve TNBC patient management. (orig.)

  5. PET Imaging of Abdominal Aortic Aneurysm with 64Cu-Labeled Anti-CD105 Antibody Fab Fragment.

    Science.gov (United States)

    Shi, Sixiang; Orbay, Hakan; Yang, Yunan; Graves, Stephen A; Nayak, Tapas R; Hong, Hao; Hernandez, Reinier; Luo, Haiming; Goel, Shreya; Theuer, Charles P; Nickles, Robert J; Cai, Weibo

    2015-06-01

    The critical challenge in abdominal aortic aneurysm (AAA) research is the accurate diagnosis and assessment of AAA progression. Angiogenesis is a pathologic hallmark of AAA, and CD105 is highly expressed on newly formed vessels. Our goal was to use (64)Cu-labeled anti-CD105 antibody Fab fragment for noninvasive assessment of angiogenesis in the aortic wall in a murine model of AAA. Fab fragment of TRC105, a mAb that specifically binds to CD105, was generated by enzymatic papain digestion and conjugated to NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) for (64)Cu labeling. The binding affinity/specificity of NOTA-TRC105-Fab was evaluated by flow cytometry and various ex vivo studies. BALB/c mice were anesthetized and treated with calcium phosphate to induce AAA and underwent weekly PET scans using (64)Cu-NOTA-TRC105-Fab. Biodistribution and autoradiography studies were also performed to confirm the accuracy of PET results. NOTA-TRC105-Fab exhibited high purity and specifically bound to CD105 in vitro. Uptake of (64)Cu-NOTA-TRC105-Fab increased from a control level of 3.4 ± 0.1 to 9.5 ± 0.4 percentage injected dose per gram (%ID/g) at 6 h after injection on day 5 and decreased to 7.2 ± 1.4 %ID/g on day 12, which correlated well with biodistribution and autoradiography studies (i.e., much higher tracer uptake in AAA than normal aorta). Of note, enhanced AAA contrast was achieved, due to the minimal background in the abdominal area of mice. Degradation of elastic fibers and highly expressed CD105 were observed in ex vivo studies. (64)Cu-NOTA-TRC105-Fab cleared rapidly through the kidneys, which enabled noninvasive PET imaging of the aorta with enhanced contrast and showed increased angiogenesis (CD105 expression) during AAA. (64)Cu-NOTA-TRC105-Fab PET may potentially be used for future diagnosis and prognosis of AAA. © 2015 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

  6. ImmunoPET of tissue factor expression in triple-negative breast cancer with a radiolabeled antibody Fab fragment.

    Science.gov (United States)

    Shi, Sixiang; Hong, Hao; Orbay, Hakan; Graves, Stephen A; Yang, Yunan; Ohman, Jakob D; Liu, Bai; Nickles, Robert J; Wong, Hing C; Cai, Weibo

    2015-07-01

    To date, there is no effective therapy for triple-negative breast cancer (TNBC), which has a dismal clinical outcome. Upregulation of tissue factor (TF) expression leads to increased patient morbidity and mortality in many solid tumor types, including TNBC. Our goal was to employ the Fab fragment of ALT-836, a chimeric anti-human TF mAb, for PET imaging of TNBC, which can be used to guide future TNBC therapy. ALT-836-Fab was generated by enzymatic papain digestion. SDS-PAGE and FACS studies were performed to evaluate the integrity and TF binding affinity of ALT-836-Fab before NOTA conjugation and (64)Cu-labeling. Serial PET imaging and biodistribution studies were carried out to evaluate the tumor targeting efficacy and pharmacokinetics in the MDA-MB-231 TNBC model, which expresses high levels of TF on the tumor cells. Blocking studies, histological assessment, as well as RT-PCR were performed to confirm TF specificity of (64)Cu-NOTA-ALT-836-Fab. ALT-836-Fab was produced with high purity, which exhibited superb TF binding affinity and specificity. Serial PET imaging revealed rapid and persistent tumor uptake of (64)Cu-NOTA-ALT-836-Fab (5.1 ± 0.5 %ID/g at 24 h post-injection; n = 4) and high tumor/muscle ratio (7.0 ± 1.2 at 24 h post-injection; n = 4), several-fold higher than that of the blocking group and tumor models that do not express significant level of TF, which was confirmed by biodistribution studies. TF specificity of the tracer was also validated by histology and RT-PCR. (64)Cu-NOTA-ALT-836-Fab exhibited prominent tissue factor targeting efficiency in MDA-MB-231 TNBC model. The use of a Fab fragment led to fast tumor uptake and good tissue/muscle ratio, which may be translated into same-day immunoPET imaging in the clinical setting to improve TNBC patient management.

  7. ImmunoPET of tissue factor expression in triple-negative breast cancer with a radiolabeled antibody Fab fragment

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Sixiang [University of Wisconsin, Materials Science Program, Madison, WI (United States); Hong, Hao; Orbay, Hakan; Yang, Yunan; Ohman, Jakob D. [University of Wisconsin, Department of Radiology, Madison, WI (United States); Graves, Stephen A.; Nickles, Robert J. [University of Wisconsin, Department of Medical Physics, Madison, WI (United States); Liu, Bai; Wong, Hing C. [Altor BioScience, Miramar, FL (United States); Cai, Weibo [University of Wisconsin, Materials Science Program, Madison, WI (United States); University of Wisconsin, Department of Radiology, Madison, WI (United States); University of Wisconsin, Department of Medical Physics, Madison, WI (United States); University of Wisconsin Carbone Cancer Center, Madison, WI (United States); University of Wisconsin, Departments of Radiology and Medical Physics, Madison, WI (United States)

    2015-07-15

    To date, there is no effective therapy for triple-negative breast cancer (TNBC), which has a dismal clinical outcome. Upregulation of tissue factor (TF) expression leads to increased patient morbidity and mortality in many solid tumor types, including TNBC. Our goal was to employ the Fab fragment of ALT-836, a chimeric anti-human TF mAb, for PET imaging of TNBC, which can be used to guide future TNBC therapy. ALT-836-Fab was generated by enzymatic papain digestion. SDS-PAGE and FACS studies were performed to evaluate the integrity and TF binding affinity of ALT-836-Fab before NOTA conjugation and {sup 64}Cu-labeling. Serial PET imaging and biodistribution studies were carried out to evaluate the tumor targeting efficacy and pharmacokinetics in the MDA-MB-231 TNBC model, which expresses high levels of TF on the tumor cells. Blocking studies, histological assessment, as well as RT-PCR were performed to confirm TF specificity of {sup 64}Cu-NOTA-ALT-836-Fab. ALT-836-Fab was produced with high purity, which exhibited superb TF binding affinity and specificity. Serial PET imaging revealed rapid and persistent tumor uptake of {sup 64}Cu-NOTA-ALT-836-Fab (5.1 ± 0.5 %ID/g at 24 h post-injection; n = 4) and high tumor/muscle ratio (7.0 ± 1.2 at 24 h post-injection; n = 4), several-fold higher than that of the blocking group and tumor models that do not express significant level of TF, which was confirmed by biodistribution studies. TF specificity of the tracer was also validated by histology and RT-PCR. {sup 64}Cu-NOTA-ALT-836-Fab exhibited prominent tissue factor targeting efficiency in MDA-MB-231 TNBC model. The use of a Fab fragment led to fast tumor uptake and good tissue/muscle ratio, which may be translated into same-day immunoPET imaging in the clinical setting to improve TNBC patient management. (orig.)

  8. Generation of human Fab antibody libraries: PCR amplification and assembly of light- and heavy-chain coding sequences.

    Science.gov (United States)

    Andris-Widhopf, Jennifer; Steinberger, Peter; Fuller, Roberta; Rader, Christoph; Barbas, Carlos F

    2011-09-01

    The development of therapeutic antibodies for use in the treatment of human diseases has long been a goal for many researchers in the antibody field. One way to obtain these antibodies is through phage-display libraries constructed from human lymphocytes. This protocol describes the construction of human Fab (fragment antigen binding) antibody libraries. In this method, the individual rearranged heavy- and light-chain variable regions are amplified separately and are linked through a series of overlap polymerase chain reaction (PCR) steps to give the final Fab products that are used for cloning.

  9. The binding parameters of radiolabelled monoclonal F (ab')2 and Fab' fragments relative to immunoglobulin G in reactions with surface-bound antigens

    International Nuclear Information System (INIS)

    Fjeld, J.G.; Nustad, K.; Michaelsen, T.E.

    1992-01-01

    The binding parameters of iodine-125-labelled intact monoclonal immunoglobulin G (IgG), F(ab') 2 and Fab' fragments were compared. The study was carried out with the two monoclonal antibodies (MoAbs) K13 and K16 specific for human Ig light chains κ and λ, respectively. When testing the 125 I-MoAbs against monodisperse polymer particles coated with the specific antigens, the K a for the F(ab') 2 fragments were similar to that for IgG, while the K a for the Fab' fragments were reduced to 10%-20% of that for IgG. The number N of effective target sites revealed with Fab' was higher than with F(ab') and IgG, presumably because less surface area is occupied by the small Fab' molecules. The immunoreactive fraction F ranged according to IgG>F(ab') 2 >Fab'. The explanation of the moderate difference between the K a of the monoclonal Fab' and the divalent IgG and F(ab') 2 was that the divalent molecules were not divalently attached to the particles. When testing the same antibody preparations against humanlymphoma cells producing Ig with light chains κ or λ, the binding results were less reliable than when particles were utilised, presumably due to antigen shedding. Different MoAbs vary in their loss of immunoreactivity due to enzymatic degradation and the radiolabelling procedure. The preparation of the radiolabelled fragments should therefore be optimized for each MoAb, and evaluation is necessary before injection. Artificial targets with a low leakage of antigen, like the monodisperse polymer particles here applied, are recommended for the in vitro evaluation of the immunoreactivity of labelled MoAb preparations. (orig.)

  10. Development of positron emission tomography probe of 64Cu-labeled anti-C-kit 12A8 Fab to measure protooncogene C-kit expression

    International Nuclear Information System (INIS)

    Yoshida, Chisato; Tsuji, Atsushi B.; Sudo, Hitomi; Sugyo, Aya; Sogawa, Chizuru; Inubushi, Masayuki; Uehara, Tomoya; Fukumura, Toshimitsu; Koizumi, Mitsuru; Arano, Yasushi; Saga, Tsuneo

    2011-01-01

    Introduction: C-kit is an important diagnostic and therapeutic target molecule for several malignancies, and c-kit-targeted drugs have been used clinically. Because abundant c-kit expression in tumors is a prerequisite for successful c-kit-targeted therapy, imaging of c-kit expression is expected to play a pivotal role in the therapeutic decision for each patient. We evaluated 64 Cu-labeled Fab of anti-c-kit antibody 12A8 as a positron emission tomography (PET) imaging probe. Methods: 111 In- or 125 I-Labeled 12A8 Fab was evaluated in vitro by cell binding, competitive inhibition and cellular internalization assays, and in vivo by biodistribution in mice bearing c-kit-expressing and -non-expressing tumors. Next, Fab fragment was labeled with the positron emitter 64 Cu and evaluated by PET. Results: Radiolabeled 12A8 Fab showed specific binding to c-kit-expressing cells with high affinity and internalized into cells after binding to c-kit on cell surface. Although tumor accumulation of [ 111 In]Fab was lower than that of [ 111 In]IgG, the faster blood clearance of [ 111 In]Fab provided higher tumor-to-blood ratio at 6 h postinjection onwards. Blood clearance of 64 Cu-labeled 12A8 Fab was slower than that of [ 111 In]Fab, but PET using [ 64 Cu]Fab clearly visualized the tumor at 6 h postinjection onwards. Conclusion: The 64 Cu-labeled 12A8 Fab could be used for c-kit-specific PET imaging and might help in selecting appropriate patients for c-kit-targeted treatments.

  11. Recombinant Human Respiratory Syncytial Virus (RSV) Monoclonal Antibody Fab is Effective Therapeutically when Introduced Directly into the Lungs of RSV-Infected Mice

    Science.gov (United States)

    Crowe, James E., Jr.; Murphy, Brian R.; Chanock, Robert M.; Williamson, R. Anthony; Barbas, Carlos F., III; Burton, Dennis R.

    1994-02-01

    Previously, recombinant human respiratory syncytial virus (RSV) monoclonal antibody Fabs were generated by antigen selection from random combinatorial libraries displayed at the tip of filamentous phage. Two such Fabs, which exhibited high binding affinity for RSV F glycoprotein (a major protective antigen), were evaluated for therapeutic efficacy in infected mice just before or at the time of peak virus replication in the lungs. Fab 19, which neutralized RSV infectivity with high efficiency in tissue culture, was effective therapeutically when delivered directly into the lungs by intranasal instillation under anesthesia. In contrast, RSV Fab 126, which failed to neutralize virus in cell culture, did not exhibit a therapeutic effect under these conditions. The amount of Fab 19 required to effect a 5000- to 12,000-fold reduction in titer of RSV in the lungs within 24 hr was rather small. In four separate experiments, a single instillation of 12.9-50 μg of RSV Fab 19 was sufficient to achieve such a reduction in pulmonary virus in a 25g mouse. The use of Fabs instead of the whole immunoglobulin molecules from which they are derived reduced the protein content of a therapeutic dose. This is important because the protein load that can be delivered effectively into the lungs is limited. The therapeutic effect of a single treatment with Fab 19 was not sustained, so that a rebound in pulmonary virus titer occurred on the 2nd day after treatment. This rebound in pulmonary RSV titer could be prevented by treating infected mice with a single dose of Fab 19 daily for 3 days. These observations suggest that human monoclonal Fabs grown in Escherichia coli may prove useful in the treatment of serious RSV disease as well as diseases caused by other viruses where replication in vivo is limited primarily to the lumenal lining of the respiratory tract.

  12. Expression of a humanized SZ-63 McAb functional recombinant Fab fragment in E. Coli

    International Nuclear Information System (INIS)

    Xia Lijun; Gu Jianming; Zhang Xiaoming; Liu Yue; Wan Haiying; Li Peixia; Ruan Changgeng

    1995-06-01

    MRNA was selected on oligo(dT)-cellulose from total RNA isolated from SZ-63 hybridoma cells by CsCl ultracentrifugation. cDNA coding for heavy and light variable regions were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR). The amplified fragments were then cloned and sequenced by the 32 P labelled sanger dideoxy-mediated chain-termination method. The nucleotides of VH and Vκ are 354 and 321 respectively, the amino acid sequence of heavy and light chain of SZ-63 were also deduced. Then, linking the variable genes of SZ-63 with human immunoglobulin γ 1 CH and κ VL genes, constructing pHEN1-63 Fab/Hu chimera for expression and transforming E. coli HB2151. The expressed chimeric SZ-63 Fab was soluble. Both ELISA and Western blot results showed the expression products could specifically bind with cross-linked fibrin and the content in expression culture was about 225 μg/L. (5 figs.)

  13. Complexes of neutralizing and non-neutralizing affinity matured Fabs with a mimetic of the internal trimeric coiled-coil of HIV-1 gp41.

    Directory of Open Access Journals (Sweden)

    Elena Gustchina

    Full Text Available A series of mini-antibodies (monovalent and bivalent Fabs targeting the conserved internal trimeric coiled-coil of the N-heptad repeat (N-HR of HIV-1 gp41 has been previously constructed and reported. Crystal structures of two closely related monovalent Fabs, one (Fab 8066 broadly neutralizing across a wide panel of HIV-1 subtype B and C viruses, and the other (Fab 8062 non-neutralizing, representing the extremes of this series, were previously solved as complexes with 5-Helix, a gp41 pre-hairpin intermediate mimetic. Binding of these Fabs to covalently stabilized chimeric trimers of N-peptides of HIV-1 gp41 (named (CCIZN363 or 3-H has now been investigated using X-ray crystallography, cryo-electron microscopy, and a variety of biophysical methods. Crystal structures of the complexes between 3-H and Fab 8066 and Fab 8062 were determined at 2.8 and 3.0 Å resolution, respectively. Although the structures of the complexes with the neutralizing Fab 8066 and its non-neutralizing counterpart Fab 8062 were generally similar, small differences between them could be correlated with the biological properties of these antibodies. The conformations of the corresponding CDRs of each antibody in the complexes with 3-H and 5-Helix are very similar. The adaptation to a different target upon complex formation is predominantly achieved by changes in the structure of the trimer of N-HR helices, as well as by adjustment of the orientation of the Fab molecule relative to the N-HR in the complex, via rigid-body movement. The structural data presented here indicate that binding of three Fabs 8062 with high affinity requires more significant changes in the structure of the N-HR trimer compared to binding of Fab 8066. A comparative analysis of the structures of Fabs complexed to different gp41 intermediate mimetics allows further evaluation of biological relevance for generation of neutralizing antibodies, as well as provides novel structural insights into immunogen

  14. Plant compounds insecticide activity against Coleoptera pests of stored products

    OpenAIRE

    MOREIRA, M.D.; PICANÇO, M.C.; BARBOSA, L.C. de A.; GUEDES, R.N.C.; CAMPOS, M.R. de; SILVA, G.A.; MARTINS, J.C.

    2008-01-01

    The objective of this work was to screen plants with insecticide activity, in order to isolate, identify and assess the bioactivity of insecticide compounds present in these plants, against Coleoptera pests of stored products: Oryzaephilus surinamensis L. (Silvanidae), Rhyzopertha dominica F. (Bostrichidae) and Sitophilus zeamais Mots. (Curculionidae). The plant species used were: basil (Ocimum selloi Benth.), rue (Ruta graveolens L.), lion's ear (Leonotis nepetifolia (L.) R.Br.), jimson weed...

  15. Sex- and Size-Related Patterns of Carrion Visitation in Necrodes littoralis (Coleoptera: Silphidae) and Creophilus maxillosus (Coleoptera: Staphylinidae).

    Science.gov (United States)

    Mądra-Bielewicz, Anna; Frątczak-Łagiewska, Katarzyna; Matuszewski, Szymon

    2017-09-01

    The estimation of postmortem interval (PMI) based on successional patterns of adult insects is largely limited, due to the lack of potential PMI markers. Sex and size of adult insects could be easily used for such estimation. In this study, sex- and size-related patterns of carrion attendance by adult insects were analyzed in Necrodes littoralis (Coleoptera: Silphidae) and Creophilus maxillosus (Coleoptera: Staphylinidae). For both species, abundance of males and females changed similarly during decomposition. A slightly female-biased sex ratio was recorded in N. littoralis. Females of N. littoralis started visiting carcasses, on average, one day earlier than males. There was a rise in size of males of N. littoralis at the end of decomposition, whereas for females of both species and males of C. maxillosus, no size-related patterns of carrion visitation were found. Current results demonstrate that size and sex of adult carrion beetles are poor indicators of PMI. © 2016 American Academy of Forensic Sciences.

  16. Estados inmaduros de Ancognatha ustulata (Coleoptera: Melolonthidae: Dynastinae: Cyclocephalini Immature stages of Ancognatha ustulata (Coleoptera: Melolonthidae: Dynastinae: Cyclocephalini

    Directory of Open Access Journals (Sweden)

    Jhon César Neita-Moreno

    2008-12-01

    Full Text Available Se describen y se ilustran por primera vez la larva de tercer estadio y la pupa de Ancognatha ustulata Burmeister, 1847 (Coleoptera: Melolonthidae: Dynastinae: Cyclocephalini. Se aportan datos sobre la biología de la especie y su distribución en Colombia. Se proporciona una clave para la identificación de las larvas de tercer estadio conocidas de las especies del género Ancognatha Erichson.The third instar larva and pupa of Ancognatha ustulata Burmeister, 1847 (Coleoptera: Melolonthidae: Dynastinae: Cyclocephalini are described and illustrated for the first time. New data on larval biology and distribution of the species in Colombia are included. A key to the known third stage larvae of Ancognatha Erichson is provided.

  17. California State University, Bakersfield Fab Lab: "Making" a Difference in Middle School Students' STEM Attitudes

    Science.gov (United States)

    Medina, Andrea Lee

    2017-01-01

    The digital fabrication lab, or Fab Lab, at California State University, Bakersfield provided a 1-week, half-day summer program for local area middle school students. The purpose of this study was to examine the effect this summer program had on their attitudes towards math and science. The theoretical framework used for this study was based on…

  18. [Molecular dynamics of immune complex of photoadduct-containing DNA with Fab-Anti-DNA antibody fragment].

    Science.gov (United States)

    Akberova, N I; Zhmurov, A A; Nevzorova, T A; Litvinov, R I

    2016-01-01

    Antibodies to DNA play an important role in the pathogenesis of autoimmune diseases. The elucidation of structural mechanisms of both the antigen recognition and the interaction of anti-DNA antibodies with DNA will help to understand the role of DNA-containing immune complexes in various pathologies and can provide a basis for new treatment modalities. Moreover, the DNA-antibody complex is an analog of specific intracellular DNA-protein interactions. In this work, we used in silico molecular dynamic simulations of bimolecular complexes of the dsDNA segment containing the Fab fragment of an anti-DNA antibody to obtain the detailed thermodynamic and structural characteristics of dynamic intermolecular interactions. Using computationally modified crystal structure of the Fab-DNA complex (PDB ID: 3VW3), we studied the equilibrium molecular dynamics of the 64M-5 antibody Fab fragment associated with the dsDNA fragment containing the thymine dimer, the product of DNA photodamage. Amino acid residues that constitute paratopes and the complementary nucleotide epitopes for the Fab-DNA construct were identified. Stacking and electrostatic interactions were found to play the main role in mediating the most specific antibody-dsDNA contacts, while hydrogen bonds were less significant. These findings may shed light on the formation and properties of pathogenic anti-DNA antibodies in autoimmune diseases, such as systemic lupus erythematosus associated with skin photosensitivity and DNA photodamage.

  19. Potent neutralization of VEGF biological activities with a fully human antibody Fab fragment directed against VEGF receptor 2

    International Nuclear Information System (INIS)

    Miao, H.-Q.; Hu, Kun; Jimenez, Xenia; Navarro, Elizabeth; Zhang, Haifan; Lu Dan; Ludwig, Dale L.; Balderes, Paul; Zhu Zhenping

    2006-01-01

    Compelling evidence suggest that vascular endothelial growth factor (VEGF) and its receptors, especially receptor 2 (VEGFR2, or kinase insert domain-containing receptor, KDR), play a critical role in angiogenesis under both physiological and pathological conditions, including cancer and angiogenic retinopathies such as age-related macular degeneration (AMD). To this end, inhibition of angiogenesis with antagonists to either VEGF or KDR has yielded significant therapeutic efficacy both in preclinical studies in animal models and in clinical trials in patients with cancer and AMD. We previously reported the identification of a high affinity, fully human anti-KDR antibody fragment, 1121B Fab, through a highly stringent affinity maturation process with a Fab originally isolated from a naive human antibody phage display library. In this study, we demonstrate that 1121B Fab is able to strongly block KDR/VEGF interaction, resulting in potent inhibition of an array of biological activities of VEGF, including activation of the receptor and its signaling pathway, intracellular calcium mobilization, and migration and proliferation of endothelial cells. Taken together, our data lend strong support to the further development of 1121B Fab fragment as an anti-angiogenesis agent in both cancer and angiogenic retinopathies

  20. Fab-dsFv: A bispecific antibody format with extended serum half-life through albumin binding.

    Science.gov (United States)

    Davé, Emma; Adams, Ralph; Zaccheo, Oliver; Carrington, Bruce; Compson, Joanne E; Dugdale, Sarah; Airey, Michael; Malcolm, Sarah; Hailu, Hanna; Wild, Gavin; Turner, Alison; Heads, James; Sarkar, Kaushik; Ventom, Andrew; Marshall, Diane; Jairaj, Mark; Kopotsha, Tim; Christodoulou, Louis; Zamacona, Miren; Lawson, Alastair D; Heywood, Sam; Humphreys, David P

    2016-10-01

    An antibody format, termed Fab-dsFv, has been designed for clinical indications that require monovalent target binding in the absence of direct Fc receptor (FcR) binding while retaining substantial serum presence. The variable fragment (Fv) domain of a humanized albumin-binding antibody was fused to the C-termini of Fab constant domains, such that the VL and VH domains were individually connected to the Cκ and CH1 domains by peptide linkers, respectively. The anti-albumin Fv was selected for properties thought to be desirable to ensure a durable serum half-life mediated via FcRn. The Fv domain was further stabilized by an inter-domain disulfide bond. The bispecific format was shown to be thermodynamically and biophysically stable, and retained good affinity and efficacy to both antigens simultaneously. In in vivo studies, the serum half-life of Fab-dsFv, 2.6 d in mice and 7.9 d in cynomolgus monkeys, was equivalent to Fab'-PEG.

  1. Immunomodulatory effects of intravenous bis-1 f(ab')(2) administration in renal-cell cancer-patients

    NARCIS (Netherlands)

    Janssen, R. A. J.; Kroesen, B. J.; Mesander, G.; Sleijfer, D. T.; The, T. Hauw; Mulder, N. H.; de Leij, L

    We report the immunomodulatory effects of an intravenous treatment with F(ab')(2) fragments of the bispecific monoclonal antibody BIS-1 during subcutaneous recombinant interleukin 2 (rIL-2) therapy of renal cell cancer (RCC) patients. BIS-1 is directed against both the CD3 antigen on T cells and the

  2. A parallel panning scheme used for selection of a GluA4-specific Fab targeting the ligand-binding domain

    DEFF Research Database (Denmark)

    Clausen, Rasmus P; Mohr, Andreas Ø; Riise, Erik

    2016-01-01

    A method for development of murine Fab fragments towards extracellular domains of a surface receptor is presented. The GluA4 ionotropic glutamate receptor is used as a model system. Recombinant GluA4 ectodomain comprising both the N-terminal domain (NTD) and the ligand-binding domain (LBD) in one...... molecule was used for immunization. A Fab-phage library was constructed and a parallel panning approach enabled selection of murine Fab fragments towards either intact ectodomain or the isolated LBD of the GluA4 receptor. One LBD-Fab (FabL9) showed exclusive selectivity for the GluA4 LBD, over a panel...... of LBDs from GluA2, GluK1, GluK2 and GluD2. Soluble FabL9 was produced in amounts suitable for characterization. Competitive ELISA and rat-brain immunoprecipitation experiments confirmed that the FabL9 epitope is conserved in the LBD and in the intact native receptor. By an alignment of GluA2 and GluA4...

  3. Radioimmunotherapy of human colon cancer xenografts by using 131I labeled-CAb1 F(ab')2

    International Nuclear Information System (INIS)

    Li Ling; Xu Huiyun; Mi Li; Bian Huijie; Qin Jun; Xiong Hua; Feng Qiang; Wen Ning; Tian Rong; Xu Liqing; Shen Xiaomei; Tang Hao; Chen Zhinan

    2006-01-01

    Purpose: Therapeutic efficacy, suitable dose, and administration times of 131 I-CAb 1 F(ab') 2 , a new monoclonal antibody therapeutics specifically directed against a cell surface-associated glycoprotein of colon cancer, were investigated in this article. Methods and Materials: In human colon cancer xenografts, 131 I-CAb 1 F(ab') 2 at the dose of 125 μCi, 375 μCi, and 1125 μCi were administrated intraperitoneally on Days 6 and 18 after implantation of HR8348 cells with CAb 1 high reactivity. Survival time and tumor growth inhibition rate were used to evaluate the efficacy and safety of 131 I-CAb 1 F(ab') 2 in treatment of colon cancer xenografts. Results: Treatment of 125, 375, and 1125 μCi 131 I-CAb1 F(ab') 2 did not significantly decrease the mean survival time of nude mice when compared with nontreated groups (p = 0.276, 0.865, 0.582, respectively). Moreover, the mean survival times of nude mice receiving 375 μCi and 1125 μCi 131 I-CAb1 F(ab') 2 were significantly longer than that of 5-FU-treated groups (p 0.018 and 0.042). Tumor growth inhibition rates of the first therapy were 35.67% and 41.37%, with corresponding 131 I-labeled antibody dosage of 375 μCi and 1125 μCi. After single attack dosage, second reinforcement therapy may rise efficacy significantly. Tumor growth inhibition rates of 125 μCi, 375 μCi, and 1125 μCi 131 I-labeled antibody on Day 20 posttherapy were 42.65%, 56.56%, and 84.41%, respectively. Histopathology examination revealed that tissue necrosis of various degrees was found in 131 I-CAb1 F(ab') 2 -treated groups. Conclusion: 131 I-CAb 1 F(ab') 2 is safe and effective for colon cancer. It may be a novel and potentially adjuvant therapeutics for colon cancer

  4. IgG4 autoantibodies against muscle-specific kinase undergo Fab-arm exchange in myasthenia gravis patients.

    Science.gov (United States)

    Koneczny, Inga; Stevens, Jo A A; De Rosa, Anna; Huda, Saif; Huijbers, Maartje G; Saxena, Abhishek; Maestri, Michelangelo; Lazaridis, Konstantinos; Zisimopoulou, Paraskevi; Tzartos, Socrates; Verschuuren, Jan; van der Maarel, Silvère M; van Damme, Philip; De Baets, Marc H; Molenaar, Peter C; Vincent, Angela; Ricciardi, Roberta; Martinez-Martinez, Pilar; Losen, Mario

    2017-02-01

    Autoimmunity mediated by IgG4 subclass autoantibodies is an expanding field of research. Due to their structural characteristics a key feature of IgG4 antibodies is the ability to exchange Fab-arms with other, unrelated, IgG4 molecules, making the IgG4 molecule potentially monovalent for the specific antigen. However, whether those disease-associated antigen-specific IgG4 are mono- or divalent for their antigens is unknown. Myasthenia gravis (MG) with antibodies to muscle specific kinase (MuSK-MG) is a well-recognized disease in which the predominant pathogenic IgG4 antibody binds to extracellular epitopes on MuSK at the neuromuscular junction; this inhibits a pathway that clusters the acetylcholine (neurotransmitter) receptors and leads to failure of neuromuscular transmission. In vitro Fab-arm exchange-inducing conditions were applied to MuSK antibodies in sera, purified IgG4 and IgG1-3 sub-fractions. Solid-phase cross-linking assays were established to determine the extent of pre-existing and inducible Fab-arm exchange. Functional effects of the resulting populations of IgG4 antibodies were determined by measuring inhibition of agrin-induced AChR clustering in C2C12 cells. To confirm the results, κ/κ, λ/λ and hybrid κ/λ IgG4s were isolated and tested for MuSK antibodies. At least fifty percent of patients had IgG4, but not IgG1-3, MuSK antibodies that could undergo Fab-arm exchange in vitro under reducing conditions. Also MuSK antibodies were found in vivo that were divalent (monospecific for MuSK). Fab-arm exchange with normal human IgG4 did not prevent the inhibitory effect of serum derived MuSK antibodies on AChR clustering in C2C12 mouse myotubes. The results suggest that a considerable proportion of MuSK IgG4 could already be Fab-arm exchanged in vivo. This was confirmed by isolating endogenous IgG4 MuSK antibodies containing both κ and λ light chains, i.e. hybrid IgG4 molecules. These new findings demonstrate that Fab-arm exchanged antibodies

  5. Elderly patients with suspected chronic digoxin toxicity: A comparison of clinical characteristics of patients receiving and not receiving digoxin-Fab.

    Science.gov (United States)

    Arbabian, Hooman; Lee, Hwee Min; Graudins, Andis

    2018-04-01

    The aim of the present study was to compare clinical features of patients with elevated serum digoxin concentrations who were treated with digoxin-Fab with those where the immunotherapy was not given by a tertiary hospital toxicology service. This was a retrospective series of patients with supratherapeutic serum digoxin concentrations referred to the toxicology service from August 2013 to October 2015. Data collected included demographics, presenting complaint, digoxin dose, other medications taken, serum digoxin, potassium and creatinine concentration on presentation and initial and post-digoxin-Fab heart rate. There were 47 referrals. Digoxin-Fab was administered in 21 cases. It was given more commonly when the heart rate was 5.0 mmol/L. Patients receiving digoxin-Fab were more likely to be on maintenance therapy with beta-blockers or calcium channel blockers (95% vs 61%; OR 13.1; 95% CI 1.5-113) and/or potassium-sparing medications (95% vs 54%; OR 17.1; 95% CI 2.0-147). They had elevated serum creatinine (76% vs 42%; OR 8.2; 95% CI 1.9-34), higher serum potassium (median: 5.1 mmol/L vs 4.2 mmol/L, P = 0.02), higher serum digoxin concentration (median: 3.5 nmol/L vs 2.3 nmol/L, P = 0.02) and pretreatment heart rate Fab. However, individual heart rate response to digoxin-Fab was variable. Digoxin-Fab was more commonly administered when heart rate was Fab was variable as patients were using other negative chronotropic medications. In symptomatic bradycardic patients on multiple heart failure medications, positive chronotropic and potassium-lowering therapies should be considered in concert with digoxin-Fab. © 2018 Australasian College for Emergency Medicine and Australasian Society for Emergency Medicine.

  6. Technetium-99m-labelled monoclonal antibody fragment (99mTc-DD-3B6/22Fab') for ovarian cancer

    International Nuclear Information System (INIS)

    Fernandes, V.; Donaghy, T.; Prabakaran, K.; Smith, S.; Clingan, P.; Bartolo, N.; Bundesen, P.; Hillyard, C.

    1998-01-01

    Full text: This study is the first to assess 99m Tc-DD-3B6/22Fab'' as an agent for detecting OC recurrence. Five informed consenting volunteers with stages 3 or 4 OC (mean age 54.2 y) and with negative skin prick test to Fab'' were recruited. All had suspicion of OC recurrence, with elevated serum tumour marker CA125, and in some, OC confirmation on CT scans. Aggressive treatment was continued so that tumour regression was possible prior to 99m Tc-DD-3B6/22Fab''scan. CA125, FBC, biochemical and clot markers were measured pre-and post-study. All received IV 600-800 MBq of 99 mTc-DD-3B6/22Fab''. Temperature, pulse and BP were monitored for one hour. Data were acquired with a single head LFOV gamma camera and HIRES collimator at 1,4,6 and 24 hours. Studies were read for OC lesions which were scored as probably normal (PN) or abnormal (PAb), definitely abnormal (DAb). The gold standard was CA125 immediately post-scan and, when possible, confirmation with CT. Of 5 patients, 1 was disease negative with 99 mTc-DD-3B6/22Fab'', CA125 suggesting remission, and 4 were positive on all modalities (Sensitivity, specificity, 100% respectively). 99 mTc-DD-3B6/22Fab'' identified 15 lesions (100% DAb). CT detected 7 lesions (71% Dab) pre- 99 mTc-DD-3B6/22Fab''. Repeat CT detected 8 (100% DAb). Correlation with CA125 levels was best with 99 mTc-DD3B6/22Fab''. No side effects or adverse change in serum markers were noted. 99 mTc-DD-3B6/22Fab'' is sensitive and specific for detecting OC recurrence

  7. Re-188 labelling of DD-3B6/22 Fab' monoclonal antibody fragment for radio immuno therapy

    International Nuclear Information System (INIS)

    Schmidt, P.F.; Smith, S.V.; Bundesen, P.

    1996-01-01

    The chemical similarity of technetium and rhenium has created much interest in the nuclear medicine field to make a 'matched pair' of radiopharmaceuticals for radioimmuno- diagnosis and therapy. Clinical trials with the 99 mTc-DD-3B6/22 Fab' has shown promise in the diagnosis of ovarian cancer. The design of the analogous therapeutic agent with rhenium-188 (155 keV γ 15 % abundant, β E max 2.1 MeV, T 1/2 17 h) is under investigation. The present study describes the approach taken for direct radiolabelling of the DD-3B6/22 Fab' with carrier-free 188 Re and its biological evaluation in balb/c and nude mice. The effect of temperature, pH and antibody concentration on the amount and rate of transchelation was also evaluated. The final product had a specific activity of 35 mCi/mg with an immunoreactive fraction of 77%. Stability of the product was assessed under various conditions: temperature, presence and absence of an inert atmosphere and presence of ascorbic acid (stabilised). Pharmacokinetics of the final product was evaluated in balb/c and nude mice transplanted with both D-dimer (+Ve) and Glycine (-Ve) beads. Results show that 188 Re DD-3B6/22 Fab' clears rapidly from the blood (α = 2.4 hr, β = 3.5 hr) and is excreted through the renal system. Localisation to subcutaneous antigen beads shows specific uptake to the D-dimer (antigen) beads was achieved within 6 h (0.23% ID) and was maintained for 24 hour post injection. Specificity to antigen implants was 5:1 (P 99m Tc DD-3B6/22 Fab' in mice. The radiolabelling procedures are congenial for therapeutic levels and hence the authors believe that the 188 Re DD-3B6/22 Fab' has some potential for use in treatment of ovarian cancer

  8. Structure of the Francisella tularensis enoyl-acyl carrier protein reductase (FabI) in complex with NAD+ and triclosan

    International Nuclear Information System (INIS)

    Mehboob, Shahila; Truong, Kent; Santarsiero, Bernard D.; Johnson, Michael E.

    2010-01-01

    Structure of the ternary complex of F. tularensis enoyl-acyl carrier protein reductase reveals the structure of the substrate binding loop whose electron density was missing in an earlier structure, and demonstrates a shift in the position of the NAD + cofactor. Enoyl-acyl carrier protein reductase (FabI) catalyzes the last rate-limiting step in the elongation cycle of the fatty-acid biosynthesis pathway and has been validated as a potential antimicrobial drug target in Francisella tularensis. The development of new antibiotic therapies is important both to combat potential drug-resistant bioweapons and to address the broader societal problem of increasing antibiotic resistance among many pathogenic bacteria. The crystal structure of FabI from F. tularensis (FtuFabI) in complex with the inhibitor triclosan and the cofactor NAD + has been solved to a resolution of 2.1 Å. Triclosan is known to effectively inhibit FabI from different organisms. Precise characterization of the mode of triclosan binding is required to develop highly specific inhibitors. Comparison of our structure with the previously determined FtuFabI structure which is bound to only NAD + reveals the conformation of the substrate-binding loop, electron density for which was missing in the earlier structure, and demonstrates a shift in the conformation of the NAD + cofactor. This shift in the position of the phosphate groups allows more room in the active site for substrate or inhibitor to bind and be better accommodated. This information will be crucial for virtual screening studies to identify novel scaffolds for development into new active inhibitors

  9. Nonspecific bowel activity in imaging inflammation with Tc-99m labelled monoclonal anti-NCA-90 Fab' fragment MN3

    International Nuclear Information System (INIS)

    Ivancevic, V.; Wolter, A.; Munz, D.L.

    2001-01-01

    Since the Tc-99m labelled monoclonal anti-NCA 90 granulocyte antibody Fab' fragment MN3 (MN3 Fab') might be of interest for imaging abdominal inflammation which could be hampered by nonspecific bowel activity, we prospectively investigated the appearance of bowel activity in MN3 Fab' imaging. Methods: Eighty consecutive patients (age range 12-85 years) referred for suspected nonabdominal, mostly musculoskeletal infection, were included. Abdominal inflammation was excluded clinically and there were no signs of inflammatory bowel disease in the patients' histories. One, 5, and 24 hours after introvenous injection of up to 1.1 GBq of MN3 Fab' planar images of the abdomen were performed. Bowel activity was graded visually using a 5-point scale. Results: The one (N = 80), 5 (N = 79), and 24 (N = 52) hour images revealed 46 (10%), 162 (34%), and 173 (55%) accumulating bowel segments, respectively, in 37 (46%), 69 (87%), and 52 (100%) patients. The mean intensity score per accumulating segment was 1.1, 1.8 and 2.7 (p = 0), respectively. Relative frequencies of appearance of the small intestine were 38%, 57%, and 21%, ileocaecal region 6%, 53%, and 48%, ascending colon 5%, 67%, and 89%, transverse colon 1%, 9%, and 69%, descending colon 8%, 15%, and 67%, and rectosigmoid 0%, 4%, and 38%, respectively. Follow-up investigations in 13 patients revealed diverging uptake patterns. Conclusion: Nonspecific bowel activity is often present in the early and almost always and more intense, in the delayed images. Early imaging at one hour after administration seems feasible, but a loss in sensitivity has to be considered. Thus, nonspecific bowel activity can be anticipated to be a pitfall in imaging abdominal inflammation with MN3 Fab'. (orig.) [de

  10. Fab Performance

    NARCIS (Netherlands)

    Ron, de A.J.; Rooda, J.E.

    2005-01-01

    To remain competitive and to boost profitability, manufacturers in capital-intensive and highly competitive industries want to maximize throughput and minimize flow time. Achieving high throughput conflicts with achieving low flow time. In order to unhide the tradeoff between throughput and flow

  11. Absorbed Doses and Risk Estimates of (211)At-MX35 F(ab')2 in Intraperitoneal Therapy of Ovarian Cancer Patients

    DEFF Research Database (Denmark)

    Cederkrantz, Elin; Andersson, Håkan; Bernhardt, Peter

    2015-01-01

    dose associated with i.p. administration of (211)At-MX35 F(ab')2. METHODS AND MATERIALS: Patients in clinical remission after salvage chemotherapy for peritoneal recurrence of ovarian cancer underwent i.p. infusion of (211)At-MX35 F(ab')2. Potassium perchlorate was given to block unwanted accumulation...... 100 MBq/L, organ equivalent doses were less than 10% of the estimated tolerance dose. CONCLUSION: Intraperitoneal (211)At-MX35 F(ab')2 treatment is potentially a well-tolerated therapy for locally confined microscopic ovarian cancer. Absorbed doses to normal organs are low, but because the effective...

  12. Family-group names in Coleoptera (Insecta)

    Science.gov (United States)

    Bouchard, Patrice; Bousquet, Yves; Davies, Anthony E.; Alonso-Zarazaga, Miguel A.; Lawrence, John F.; Lyal, Chris H. C.; Newton, Alfred F.; Reid, Chris A. M.; Schmitt, Michael; Ślipiński, S. Adam; Smith, Andrew B. T.

    2011-01-01

    Abstract We synthesize data on all known extant and fossil Coleoptera family-group names for the first time. A catalogue of 4887 family-group names (124 fossil, 4763 extant) based on 4707 distinct genera in Coleoptera is given. A total of 4492 names are available, 183 of which are permanently invalid because they are based on a preoccupied or a suppressed type genus. Names are listed in a classification framework. We recognize as valid 24 superfamilies, 211 families, 541 subfamilies, 1663 tribes and 740 subtribes. For each name, the original spelling, author, year of publication, page number, correct stem and type genus are included. The original spelling and availability of each name were checked from primary literature. A list of necessary changes due to Priority and Homonymy problems, and actions taken, is given. Current usage of names was conserved, whenever possible, to promote stability of the classification. New synonymies (family-group names followed by genus-group names): Agronomina Gistel, 1848 syn. nov. of Amarina Zimmermann, 1832 (Carabidae), Hylepnigalioini Gistel, 1856 syn. nov. of Melandryini Leach, 1815 (Melandryidae), Polycystophoridae Gistel, 1856 syn. nov. of Malachiinae Fleming, 1821 (Melyridae), Sclerasteinae Gistel, 1856 syn. nov. of Ptilininae Shuckard, 1839 (Ptinidae), Phloeonomini Ádám, 2001 syn. nov. of Omaliini MacLeay, 1825 (Staphylinidae), Sepedophilini Ádám, 2001 syn. nov. of Tachyporini MacLeay, 1825 (Staphylinidae), Phibalini Gistel, 1856 syn. nov. of Cteniopodini Solier, 1835 (Tenebrionidae); Agronoma Gistel 1848 (type species Carabus familiaris Duftschmid, 1812, designated herein) syn. nov. of Amara Bonelli, 1810 (Carabidae), Hylepnigalio Gistel, 1856 (type species Chrysomela caraboides Linnaeus, 1760, by monotypy) syn. nov. of Melandrya Fabricius, 1801 (Melandryidae), Polycystophorus Gistel, 1856 (type species Cantharis aeneus Linnaeus, 1758, designated herein) syn. nov. of Malachius Fabricius, 1775 (Melyridae), Sclerastes

  13. Family-Group Names In Coleoptera (Insecta

    Directory of Open Access Journals (Sweden)

    Patrice Bouchard

    2011-04-01

    Full Text Available We synthesize data on all known extant and fossil Coleoptera family-group names for the first time. A catalogue of 4887 family-group names (124 fossil, 4763 extant based on 4707 distinct genera in Coleoptera is given. A total of 4492 names are available, 183 of which are permanently invalid because they are based on a preoccupied or a suppressed type genus. Names are listed in a classification framework. We recognize as valid 24 superfamilies, 211 families, 541 subfamilies, 1663 tribes and 740 subtribes. For each name, the original spelling, author, year of publication, page number, correct stem and type genus are included. The original spelling and availability of each name were checked from primary literature. A list of necessary changes due to Priority and Homonymy problems, and actions taken, is given. Current usage of names was conserved, whenever possible, to promote stability of the classification. New synonymies (family-group names followed by genus-group names: Agronomina Gistel, 1848 syn. n. of Amarina Zimmermann, 1832 (Carabidae, Hylepnigalioini Gistel, 1856 syn. n. of Melandryini Leach, 1815 (Melandryidae, Polycystophoridae Gistel, 1856 syn. n. of Malachiinae Fleming, 1821 (Melyridae, Sclerasteinae Gistel, 1856 syn. n. of Ptilininae Shuckard, 1839 (Ptinidae, Phloeonomini Ádám, 2001 syn. n. of Omaliini MacLeay, 1825 (Staphylinidae, Sepedophilini Ádám, 2001 syn. n. of Tachyporini MacLeay, 1825 (Staphylinidae, Phibalini Gistel, 1856 syn. n. of Cteniopodini Solier, 1835 (Tenebrionidae; Agronoma Gistel 1848 (type species Carabus familiaris Duftschmid, 1812, designated herein syn. n. of Amara Bonelli, 1810 (Carabidae, Hylepnigalio Gistel, 1856 (type species Chrysomela caraboides Linnaeus, 1760, by monotypy syn. n. of Melandrya Fabricius, 1801 (Melandryidae, Polycystophorus Gistel, 1856 (type species Cantharis aeneus Linnaeus, 1758, designated herein syn. n. of Malachius Fabricius, 1775 (Melyridae, Sclerastes Gistel, 1856 (type species

  14. Effective EUVL mask cleaning technology solutions for mask manufacturing and in-fab mask maintenance

    Science.gov (United States)

    Dietze, Uwe; Dress, Peter; Waehler, Tobias; Singh, Sherjang; Jonckheere, Rik; Baudemprez, Bart

    2011-03-01

    Extreme Ultraviolet Lithography (EUVL) is considered the leading lithography technology choice for semiconductor devices at 16nm HP node and beyond. However, before EUV Lithography can enter into High Volume Manufacturing (HVM) of advanced semiconductor devices, the ability to guarantee mask integrity at point-of-exposure must be established. Highly efficient, damage free mask cleaning plays a critical role during the mask manufacturing cycle and throughout the life of the mask, where the absence of a pellicle to protect the EUV mask increases the risk of contamination during storage, handling and use. In this paper, we will present effective EUVL mask cleaning technology solutions for mask manufacturing and in-fab mask maintenance, which employs an intelligent, holistic approach to maximize Mean Time Between Cleans (MBTC) and extend the useful life span of the reticle. The data presented will demonstrate the protection of the capping and absorber layers, preservation of pattern integrity as well as optical and mechanical properties to avoid unpredictable CD-linewidth and overlay shifts. Experiments were performed on EUV blanks and pattern masks using various process conditions. Conditions showing high particle removal efficiency (PRE) and minimum surface layer impact were then selected for durability studies. Surface layer impact was evaluated over multiple cleaning cycles by means of UV reflectivity metrology XPS analysis and wafer prints. Experimental results were compared to computational models. Mask life time predictions where made using the same computational models. The paper will provide a generic overview of the cleaning sequence which yielded best results, but will also provide recommendations for an efficient in-fab mask maintenance scheme, addressing handling, storage, cleaning and inspection.

  15. A KAS2 cDNA complements the phenotypes of the Arabidopsis fab1 mutant that differs in a single residue bordering the substrate binding pocket

    DEFF Research Database (Denmark)

    Carlsson, A.S.; LaBrie, S.T.; Kinney, A.J.

    2002-01-01

    The fab1 mutant of Arabidopsis is partially deficient in activity of ß-ketoacyl-[acyl carrier protein] synthase II (KAS II). This defect results in increased levels of 16 : 0 fatty acid and is associated with damage and death of the mutants at low temperature. Transformation of fab1 plants with a c......DNA from Brassica napus encoding a KAS II enzyme resulted in complementation of both mutant phenotypes. The dual complementation by expression of the single gene proves that low-temperature damage is a consequence of altered membrane unsaturation. The fab1 mutation is a single nucleotide change...... chain to bend. For functional analysis the equivalent Leu207Phe mutation was introduced into the fabB gene encoding the E. coli KAS I enzyme. Compared to wild-type, the Leu207Phe protein showed a 10-fold decrease in binding affinity for the fatty acid substrate, exhibited a modified behavior during size...

  16. Staphylococcus aureus but not Listeria monocytogenes adapt to triclosan and adaptation correlates with increased fabI expression and agr deficiency

    DEFF Research Database (Denmark)

    Nielsen, Lene Nørby; Larsen, Marianne Halberg; Skovgaard, Sissel

    2013-01-01

    was initially 4 mg/L and remained unaltered by the exposure. The adapted S. aureus isolates retained normal colony size but displayed increased expression of fabI encoding an essential enzyme in bacterial fatty acid synthesis. Also, they displayed decreased or no expression of the virulence associated agr......C of the agr quorum sensing system. While most adapted strains of USA300 carried mutations in fabI, none of the adapted strains of 8325-4 did. Conclusions. Adaptability to triclosan varies substantially between Gram positive human pathogens. S. aureus displayed an intrinsically lower MIC for triclosan compared...... to L. monocytogenes but was easily adapted leading to the same MIC as L. monocytogenes. Even though all adapted S. aureus strains over-expressed fabI and eliminated expression of the agr quorum sensing system, adaptation in USA300 involved fabI mutations whereas this was not the case for 8325-4. Thus...

  17. Anti-colchicine Fab fragments prevent lethal colchicine toxicity in a porcine model: a pharmacokinetic and clinical study.

    Science.gov (United States)

    Eddleston, Michael; Fabresse, Nicolas; Thompson, Adrian; Al Abdulla, Ibrahim; Gregson, Rachael; King, Tim; Astier, Alain; Baud, Frederic J; Clutton, R Eddie; Alvarez, Jean-Claude

    2018-08-01

    Colchicine poisoning is commonly lethal. Colchicine-specific Fab fragments increase rat urinary colchicine clearance and have been associated with a good outcome in one patient. We aimed to develop a porcine model of colchicine toxicity to study the pharmacokinetics and efficacy of ovine Fab. A Göttingen minipig critical care model was established and serial blood samples taken for colchicine and Fab pharmacokinetics, clinical chemistry, and haematology. Animals were euthanised when the mean arterial pressure fell below 45 mmHg without response to vasopressor, or at study completion. Initial studies indicated that oral dosing produced variable pharmacokinetics and time-to-euthanasia. By contrast, intravenous infusion of 0.25 mg/kg colchicine over 1 h produced reproducible pharmacokinetics (AUC 0-20 343 [SD = 21] µg/L/h), acute multi-organ injury, and cardiotoxicity requiring euthanasia a mean of 22.5 (SD = 3.2) h after dosing. A full-neutralising equimolar Fab dose given 6 h after the infusion (50% first hour, 50% next 6 h [to reduce renal-loss of unbound Fab]) produced a 7.35-fold increase in plasma colchicine (AUC 0-20 2,522 [SD = 14] µg/L/h), and removed all free plasma colchicine, but did not prevent toxicity (euthanasia at 29.1 [SD = 3.4] h). Earlier administration over 1 h of the full-neutralising dose, 1 or 3 h after the colchicine, produced a 12.9-fold (AUC 0-20 4,433 [SD = 607] µg/L/h) and 6.0-fold (AUC 0-20 2,047 [SD = 51] µg/L/h) increase in plasma colchicine, respectively, absence of free plasma colchicine until 20 h, and survival to study end without marked cardiotoxicity. Colchicine-specific Fab given early, in equimolar dose, bound colchicine, eliciting its movement into the blood, and preventing severe toxicity. Clinical studies are now needed to determine how soon this antidote must be given to work in human poisoning.

  18. Preparation, biodistribution and dosimetry of copper-64-labeled anti-colorectal carcinoma monoclonal antibody fragments 1A3-F(ab')2

    International Nuclear Information System (INIS)

    Anderson, C.J.; Schwarz, S.W.; Connett, J.M.

    1995-01-01

    Antibody fragments labeled with a radiometal using bifunctional chelates generally undergo renal clearance followed by trapping of the metabolites, leading to high radiation doses to the kidneys. Copper-64-labeled BAT-2IT-1A3-F(ab') 2 was recently reported to accumulate in colorectal tumors in an animal model, however, kidney uptake was also high. In this study, the preparation of 64 Cu-BAT-2IT-1A3-F(ab') 2 was optimized to reduce the renal uptake. The bifunctional chelate 6-bromoacetamidobenzyl-1,4,8,11-tetraazacyclotetradecane-N,N',N double-prime,N'double-prime-tetraacetic acid (BAT) was conjugated to 1A3-F(ab') 2 using the linking agent 2-iminothiolane (2IT). The conjugation reaction produced 20% of a lower molecular weight impurity found to be TETA-1A3-Fab'. The conjugation procedure was optimized to include FPLC purification of the BAT-2IT-1A3-F(ab') 2 from TETA-1A3-Fab' after conjugation prior to labeling with 64 Cu. The biodistribution of 64 Cu-labeled FPLC-purified and unpurified conjugates was determined in normal Sprague-Dawley rats and tumor-bearing Golden Syrian hamsters. Human absorbed doses were calculated from rat biodistribution data and PET imaging of a baboon. Upon FPLC purification of the BAT-2IT-1A3-F(ab') 2 , the immunoreactivity of 64 Cu-labeled 1A3-F(ab') 2 was significantly improved over that of non-FPLC-purified 64 Cu-BAT-2IT-1A3-F(ab') 2 , and the kidney uptake was decreased in normal rats. The biodistribution in hamsters showed some improvement in both tumor uptake and kidney clearance with FPLC-purified 64 Cu-BAT-2IT-1A3-F(ab') 2 .The improved dosimetry of 64 Cu-labeled FPLC purified BAT-2IT-1A3-F(ab') 2 should more readily allow this agent to be investigated clinically to image colorectal cancer using PET. 33 refs., 7 figs., 3 tabs

  19. Scymnus camptodromus (Coleoptera: Coccinellidae) larval development and predation of hemlock woolly adelgid (Hemiptera: Adelgidae)

    Science.gov (United States)

    Samita Limbu; Melody A. Keena; David Long; Nancy Ostiguy; Kelli. Hoover

    2015-01-01

    Development time and prey consumption of Scymnus (Neopullus) camptodromus Yu and Liu (Coleoptera: Coccinellidae) larvae by instar, strain, and temperature were evaluated. S. camptodromus, a specialist predator of hemlock woolly adelgid Adelges tsugae (Annand) (Hemiptera:...

  20. First record of Molorchus minor minor (Linnaeus (Coleoptera, Cerambycidae in Brazil

    Directory of Open Access Journals (Sweden)

    Ubirajara R. Martins

    2015-03-01

    Full Text Available Molorchus minor minor (Linnaeus (Coleoptera, Cerambycidae is recorded for the first time in Brazil (Bahia. It was originally described from Europe and is currently widely distributed in that continent and Asia.

  1. Aggregation pheromone compounds of the black larder beetle Dermestes haemorrhoidalis Kuster (Coleoptera: Dermestidae)

    NARCIS (Netherlands)

    Korada, R.R.; Griepink, F.C.

    2011-01-01

    Gas chromatography with simultaneous flame ionisation and electroantennographic detection (GCEAD) and gas chromatography with mass spectrometry analysis (GCMS) of abdominal extracts of adult male Dermestes haemorrhoidalis Kuster (Coleoptera: Dermestidae) revealed the presence of

  2. A predictive distribution model for Graphoderus bilineatus in the Netherlands (Coleoptera: Dytiscidae).

    NARCIS (Netherlands)

    Sierdsema, H.; Cuppen, J.G.M.

    2006-01-01

    Een voorspellend verspreidingsmodel voor de gestreepte waterroofkever Graphoderus bilineatus in Nederland (Coleoptera: Dytiscidae) Op verzoek van de Provincie Zuid-Holland en het Ministerie van lnv is het huidige voorkomen en de biotoopvoorkeur van de gestreepte waterroofkever Graphoderus bilineatus

  3. Illustrated guide to the emerald ash borer, Agrilus planipennis Fairmaire and related species (Coleoptera, Buprestidae)

    Science.gov (United States)

    The 33 species of Agrilus (Coleoptera: Buprestidae) hypothesized to be most closely related to Agrilus planipennis Fairmaire (the emerald ash borer), are described and illustrated. Morphology (adults and immatures), biology, distribution, detailed taxonomic history and systematics are presented fo...

  4. Two new fossil species of Cryptocephalus Geoffroy (Coleoptera: Chrysomelidae) from Baltic and Dominican Amber

    Science.gov (United States)

    Two new species of Cryptocephalus Geoffroy (Coleoptera: Chrysomelidae) are described and illustrated from fossil resin: Cryptocephalus groehni sp. nov (Baltic amber) and Cryptocephalus kheelorum sp. nov. (Dominican amber). These are the first described species of Cryptocephalinae from fossil resin. ...

  5. Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes.

    Science.gov (United States)

    Bondt, Albert; Rombouts, Yoann; Selman, Maurice H J; Hensbergen, Paul J; Reiding, Karli R; Hazes, Johanna M W; Dolhain, Radboud J E M; Wuhrer, Manfred

    2014-11-01

    The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. In contrast, little is known about Fab-linked N-glycosylation, carried by ∼ 20% of IgG. Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. Additionally, we observed several changes during pregnancy and after delivery. Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. Fc glycosylation changes were similar to results described before, with increased galactosylation and sialylation and decreased bisection during pregnancy. We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. Relative stabilities of IgG1 and IgG4 Fab domains: Influence of the light–heavy interchain disulfide bond architecture

    Science.gov (United States)

    Heads, James T; Adams, Ralph; D'Hooghe, Lena E; Page, Matt J T; Humphreys, David P; Popplewell, Andrew G; Lawson, Alastair D; Henry, Alistair J

    2012-01-01

    The stability of therapeutic antibodies is a prime pharmaceutical concern. In this work we examined thermal stability differences between human IgG1 and IgG4 Fab domains containing the same variable regions using the thermofluor assay. It was found that the IgG1 Fab domain is up to 11°C more stable than the IgG4 Fab domain containing the same variable region. We investigated the cause of this difference with the aim of developing a molecule with the enhanced stability of the IgG1 Fab and the biological properties of an IgG4 Fc. We found that replacing the seven residues, which differ between IgG1 CH1 and IgG4 CH1 domains, while retaining the native IgG1 light-heavy interchain disulfide (L–H) bond, did not affect thermal stability. Introducing the IgG1 type L–H interchain disulfide bond (DSB) into the IgG4 Fab resulted in an increase in thermal stability to levels observed in the IgG1 Fab with the same variable region. Conversely, replacement of the IgG1 L–H interchain DSB with the IgG4 type L–H interchain DSB reduced the thermal stability. We utilized the increased stability of the IgG1 Fab and designed a hybrid antibody with an IgG1 CH1 linked to an IgG4 Fc via an IgG1 hinge. This construct has the expected biophysical properties of both the IgG4 Fc and IgG1 Fab domains and may therefore be a pharmaceutically relevant format. PMID:22761163

  7. Immunoglobulin G (IgG) Fab Glycosylation Analysis Using a New Mass Spectrometric High-throughput Profiling Method Reveals Pregnancy-associated Changes*

    Science.gov (United States)

    Bondt, Albert; Rombouts, Yoann; Selman, Maurice H. J.; Hensbergen, Paul J.; Reiding, Karli R.; Hazes, Johanna M. W.; Dolhain, Radboud J. E. M.; Wuhrer, Manfred

    2014-01-01

    The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. In contrast, little is known about Fab-linked N-glycosylation, carried by ∼20% of IgG. Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. Additionally, we observed several changes during pregnancy and after delivery. Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. Fc glycosylation changes were similar to results described before, with increased galactosylation and sialylation and decreased bisection during pregnancy. We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. PMID:25004930

  8. Radioimmunoimaging of human breast carcinoma xenografts in nude mouse model with 111In-labeled new monoclonal antibody EBA-1 and F(ab')2 fragments

    International Nuclear Information System (INIS)

    Yemul, Shrishailam; Leon, J.A.; Pozniakoff, Ted; Esser, P.D.; Estabrook, Alison; Met-Path Inc., Teterboro, NJ

    1993-01-01

    Radioimmunoimaging characteristics of a new monoclonal antibody EBA-1 and its F(ab') 2 fragments utilizing nu/nu mice bearing human breast carcinoma xenografts are described. 111 In-DPTA conjugates of EBA-1 localized with tumor/blood ratios of 0.99 ± 0.10 (P 2 radioconjugates at 48 h. These results suggest that EBA-1 and its F(ab') 2 might be useful reagents in radioimmunoimaging and radioimmunotherapy. (author)

  9. Passive immunotherapy for influenza A H5N1 virus infection with equine hyperimmune globulin F(ab'2 in mice

    Directory of Open Access Journals (Sweden)

    Li Yanbin

    2006-03-01

    Full Text Available Abstract Background Avian influenza virus H5N1 has demonstrated considerable pandemic potential. Currently, no effective vaccines for H5N1 infection are available, so passive immunotherapy may be an alternative strategy. To investigate the possible therapeutic effect of antibody against highly pathogenic H5N1 virus on a mammal host, we prepared specific equine anti-H5N1 IgGs from horses vaccinated with inactivated H5N1 virus, and then obtained the F(ab'2 fragments by pepsin digestion of IgGs. Methods The horses were vaccinated with inactivated H5N1 vaccine to prepare anti-H5N1 IgGs. The F(ab'2 fragments were purified from anti-H5N1 hyperimmune sera by a protocol for 'enhanced pepsin digestion'. The protective effect of the F(ab'2 fragments against H5N1 virus infection was determined in cultured MDCK cells by cytopathic effect (CPE assay and in a BALB/c mouse model by survival rate assay. Results By the protocol for 'enhanced pepsin digestion', total 16 g F(ab'2 fragments were finally obtained from one liter equine antisera with the purity of over 90%. The H5N1-specific F(ab'2 fragments had a HI titer of 1:1024, and the neutralization titre of F(ab'2 reached 1: 2048. The in vivo assay showed that 100 μg of the F(ab'2 fragments could protect BALB/c mice infected with a lethal dose of influenza H5N1 virus. Conclusion The availability of highly purified H5N1-specific F(ab'2 fragments may be promising for treatment of influenza H5N1 infection. Our work has provided experimental support for the application of the therapeutic equine immunoglobulin in future large primate or human trials.

  10. Comparative study on the ability of IgG and F(ab')2 antivenoms to neutralize lethal and myotoxic effects induced by Micrurus nigrocinctus (coral snake) venom

    OpenAIRE

    León Montero, Guillermo; Stiles, Bradley G.; Alape Girón, Alberto; Rojas Céspedes, Gustavo; Gutiérrez, José María

    1999-01-01

    A comparative study was performed on the ability of IgG and F(ab')2 antivenoms to neutralize lethal and myotoxic activities of Micrurus nigrocinctus venom. Both antivenoms were adjusted to a similar neutralizing potency in experiments where venom and antivenoms were preincubated prior to injection. No significant differences were observed between IgG and F(ab')2 antivenoms concerning neutralization of lethal effect in rescue experiments, i.e., when antivenom was administered intravenously aft...

  11. Primer registro de Hippodamia variegata (Goeze 1777) (Coleoptera: Coccinellidae) en el Perú.

    OpenAIRE

    Bustamante-Navarrete, Abdhiel; Marquina-Montesinos, Edgar Luis; Elme-Tumpay, Araseli

    2017-01-01

    First report of Hippodamia variegata (Goeze 1777) (Coleoptera: Coccinellidae) in Peru. The first record of Hippodamia variegata (Goeze 1777) (Coleoptera: Coccinellidae) in Peru is reported, which also constitutes the highest record of the species. This introduced species is already established in the rest of the continent, but its presence in Peru is confirmed in this contribution. A distribution map and a key to distinguish the two species of this genus present in Peru are presented.

  12. Biologi Hama Kumbang Penggerek Pucuk Kelapa Sawit (Oryctes rhinoceros L.) (Coleoptera: Scarabaeidae) Di Rumah Kassa

    OpenAIRE

    Sejahtra, Ahmad

    2011-01-01

    Ahmad Sejahtra, "BIOLOGY SHOOT BORERS BEETLE PEST OF COCONUT PALM (Oryctes rhinoceros L.) (Coleoptera: Scarabaeidae) AT KASSA HOUSE", Under supervised by Ms Marheni and Ms Fatima Zahara. This study aims to determine biological pest of oil palm bud weevil (Oryctes rhinoceros L.) (Coleoptera: Scarabaeidae) at home kassa. The experiment was conducted on people's plantations in PERUMNAS Simalingkar starting from September 2010 until April 2011. Research using observational methods for this resear...

  13. Localization of mammary tumors in vivo with 131I-labeled Fab fragments of antibodies against mouse mammary epithelial (MME) antigens

    International Nuclear Information System (INIS)

    Wilbanks, T.; Peterson, J.A.; Miller, S.; Kaufman, L.; Ortendahl, D.; Ceriani, R.L.

    1981-01-01

    The Fab fragments of antibodies against cell-type-specific surface antigens of mouse mammary epithelial cells (MME-antigens) were used to localize mammary tumors successfully. The radioiodine-labeled anti-MME (Fab) was injected into mice carrying simulated mammary metastases, and after 24 hours the amount of label per gram of excised tissue was several times greater in the tumor than in liver, brain, lung, or muscle. Kidney showed considerable accumulation of label but this appeared to be nonspecific. Kinetic studies revealed a rapid elimination of labeled Fab in the urine with only 1% of the injected dose remaining in the entire blood pool after 24 hours. Wit a high-purity germanium camera, mammary tumors were clearly located ty the 131 I-labeled anti-MME (Fab), and normalization to /sup 99m/Tc-pertechnetate distribution in the animal increased the specificity. The density of 131 I-label was fourfold greater over the mammary tumor than over comparable areas of the mouse. No accumulation of 131 I-anti-MME (Fab) was observed in nonmammary tumors nor in mammary tumors when labeled nonspecific Fab was used. An analogous system using an antihuman mammary epithelial antiserum is being developed for localization of breast metastases in humans

  14. Extracorporeal life support and digoxin-specific Fab fragments for successful management of Taxus baccata intoxication with low output and ventricular arrhythmia.

    Science.gov (United States)

    Farag, Mina; Badowski, Dominika; Koschny, Ronald; Skopp, Gisela; Brcic, Andreas; Szabo, Gabor B

    2017-12-01

    Yew plants are evergreen shrubs which are widely spread throughout the northern hemisphere. Taxane alkaloid derivatives, mainly taxine B, represent the main toxins of Taxus baccata and are highly cardiotoxic. Due to the lack of randomized clinical trials, case reports on accidental or suicidal yew intoxications build the only source of knowledge of clinical treatment options. We report the case of a suicidal yew ingestion admitted to our hospital under prolonged cardiopulmonary resuscitation due to pulseless electrical activity. Extra-corporeal life support (ECLS) was established to maintain adequate organ perfusion. Repeated administration of digoxin-specific Fab antibody fragments, which cross-react with taxine, was associated with an immediate conversion from asystole to broad-complex bradycardia and a gradual normalization of the electrocardiogram (ECG). This was paralleled by a recovery of the cardiac function and weaning from the ECLS. The taxine metabolite 3,5-dimethoxyphenol could be detected by mass spectrometry before but not after the first Fab-fragment treatment. In contrast, the total amount of taxine (including the neutralized, Fab fragment-bound fraction) was increased after each Fab fragment administration, suggesting an accumulation of neutralized, since antibody-bound taxine in the blood by anti-digoxin Fab fragments. In conclusion, the successful clinical course of this case suggests a benefit of an early anti-digoxin Fab-fragment administration for the treatment of yew intoxication. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Studies of nontarget-mediated distribution of human full-length IgG1 antibody and its FAb fragment in cardiovascular and metabolic-related tissues.

    Science.gov (United States)

    Davidsson, Pia; Söderling, Ann-Sofi; Svensson, Lena; Ahnmark, Andrea; Flodin, Christine; Wanag, Ewa; Screpanti-Sundqvist, Valentina; Gennemark, Peter

    2015-05-01

    Tissue distribution and pharmacokinetics (PK) of full-length nontargeted antibody and its antigen-binding fragment (FAb) were evaluated for a range of tissues primarily of interest for cardiovascular and metabolic diseases. Mice were intravenously injected with a dose of 10 mg/kg of either human IgG1or its FAb fragment; perfused tissues were collected at a range of time points over 3 weeks for the human IgG1 antibody and 1 week for the human FAb antibody. Tissues were homogenized and antibody concentrations were measured by specific immunoassays on the Gyros system. Exposure in terms of maximum concentration (Cmax ) and area under the curve was assessed for all nine tissues. Tissue exposure of full-length antibody relative to plasma exposure was found to be between 1% and 10%, except for brain (0.2%). Relative concentrations of FAb antibody were the same, except for kidney tissue, where the antibody concentration was found to be ten times higher than in plasma. However, the absolute tissue uptake of full-length IgG was significantly higher than the absolute tissue uptake of the FAb antibody. This study provides a reference PK state for full-length whole and FAb antibodies in tissues related to cardiovascular and metabolic diseases that do not include antigen or antibody binding. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

  16. Comparative study of intact A7 MoAc and F(ab')2 fragments for radioimmunoimaging of human colon cancer in nude mice

    International Nuclear Information System (INIS)

    Kojima, Shuji; Suzuki, Naomi; Shimura, Noriko; Kubodera, Akiko; Kubota, Kazuhiko; Yamaguchi, Toshiharu; Takahashi, Toshio; Oyamada, Hiyoshimaru

    1993-01-01

    Differences of pharmacokinetics and tumor imaging ability between intact monoclonal antibody A7 (A7 MoAb) and F(ab) 2 fragments were studied in human colon cancer (LS-174T)-bearing nude mice. The authors examined the yield and the immunoreactivity of F(ab) 2 fragments after treatment with ficin as a function of time. The yield of F(ab) 2 fragments reached about 50% after ficin treatment for 8 h, and the F(ab) 2 retained about 80% of the immunoreactivity of the corresponding MoAb. Longer digestion with ficin produced smaller fragments (less than 92 kDa) with a lower yield and most of the immunoreactivity was lost. In pharmacokinetics studies, the F(ab') 2 was preferentially taken up by the tumor, cleared more rapidly from the blood circulation and seemed to have less non-specific tissue binding than intact A7 MoAb. The tumor image obtained at an early time using 131 I-F(ab') 2 was much superior in quality to that with intact 131 I-A7 MoAb. The use of F(ab') 2 fragments may be effective for tumor diagnosis and therapy. (author)

  17. Development of positron emission tomography imaging by 64Cu-labeled Fab for detecting ERC/mesothelin in a mesothelioma mouse model.

    Science.gov (United States)

    Yoshida, Chisato; Sogawa, Chizuru; Tsuji, Atsushi B; Sudo, Hitomi; Sugyo, Aya; Uehara, Tomoya; Hino, Okio; Yoshii, Yukie; Fujibayashi, Yasuhisa; Fukumura, Toshimitsu; Koizumi, Mitsuru; Arano, Yasushi; Saga, Tsuneo

    2010-05-01

    Malignant mesothelioma is a highly aggressive form of cancer. Curative surgery is the only effective therapy for mesothelioma, and therefore early diagnosis is important. However, early diagnosis is difficult using current diagnostic imaging techniques, and a new imaging method for early diagnosis is urgently required. We evaluated the affinity of radiolabeled monoclonal antibodies to the C-terminal fragment of ERC/mesothelin for this purpose. In-labeled or I-labeled IgG against C-terminal fragment of ERC and its Fab fragment were evaluated in vitro by cell binding, competitive inhibition, and cellular internalization assays, and in vivo by biodistribution in mice bearing ERC-expressing tumors. Next, the Fab fragment was labeled with the positron emitter Cu and evaluated by positron emission tomography (PET). Radiolabeled IgG and Fab showed specific binding to ERC-expressing mesothelioma cells with high affinity. Both radiolabeled IgG and Fab internalized into cells after binding to ERC on the cell surface. In-labeled IgG accumulated in ERC-expressing tumors and resulted in a moderate tumor-to-blood ratio at 4 days after injection. Furthermore, PET using Cu-labeled Fab visualized the tumor at 6 h after injection. Cu-labeled Fab can be useful for ERC-specific PET imaging, and can thus facilitate improved diagnosis of patients with early-stage mesothelioma.

  18. The tribe Phanaeini (Coleoptera: Scarabaeidae: Scarabaeinae in Peru

    Directory of Open Access Journals (Sweden)

    Luis Figueroa

    2014-10-01

    Full Text Available We examine the occurrence of the tribe Phanaeini (Coleoptera: Scarabaeidae: Scarabaeinae in Peru based on the collection at Museo de Historia Natural of the Universidad Nacional Mayor de San Marcos and on data provided in literature. Each species is presented with citations of its diagnosis, distribution and related comments. Peruvian Phanaeini includes 30 species in nine genera: Coprophanaeus, Dendropaemon, Gromphas, Oruscatus, Oxysternon, Phanaeus, Sulcophanaeus, Tetramereia and Megatharsis. Oruscatus davus is the only species distributed in the high Andes; Phanaeus lunaris and P. achilles occur in the northern arid zone shared by Peru and Ecuador; the remaining species are Amazonian.

  19. Papel dos besouros (Insecta, Coleoptera na Entomologia Forense

    Directory of Open Access Journals (Sweden)

    Wellington Emanuel dos Santos

    2014-12-01

    Full Text Available Esse trabalho apresenta uma revisão do papel que os besouros (Insecta, Coleoptera desempenham na Entomologia Forense. Discussões sobre ocorrência em cadáveres humanos e carcaças animais, estimativas de Intervalo Pós-Morte (IPM, estudos realizados no Brasil e em outros países, principais famílias de importância forense e aspectos biológicos, ecológicos e biogeográficos das espécies são apresentadas.

  20. Checklist of beetles (Coleoptera of Canada and Alaska. Second edition

    Directory of Open Access Journals (Sweden)

    Yves Bousquet

    2013-12-01

    Full Text Available All 8237 species-group taxa of Coleoptera known to occur in Canada and Alaska are recorded by province/territory or state, along with their author(s and year of publication, in a classification framework. Only presence of taxa in each Canadian province or territory and Alaska is noted. Labrador is considered a distinct geographical entity. Adventive and Holarctic species-group taxa are indicated. References to pertinent identification keys are given under the corresponding supraspecific taxa in the data archive.

  1. Ground beetles (Coleoptera, Carabidae agrocenoses of spring and winter wheat

    Directory of Open Access Journals (Sweden)

    Luboš Purchart

    2005-01-01

    Full Text Available On two monitoring areas of the Central Institute for Supervising and Testing in Agriculture (ÚKZÚZ loaded with risk elements we carried out investigations of beetles of the family Carabidae (Coleoptera in agricultural stands of winter and spring wheat. The focus of the present study is on synecological characteristics and in some extent on the impact of agricultural practise on the population and seasonal dynamics of the most important representatives of ground beetles. This paper precedes the following article aimed to contents of heavy metals in ground beetles.

  2. Revision of the genus Endochilus Weise (Coleoptera: Coccinellidae: Chilocorini)

    Science.gov (United States)

    Łączyński, Piotr; Tomaszewska, Wioletta

    2014-01-01

    Abstract The members of the endemic African genus EndochilusWeise, 1898 (Coleoptera: Coccinellidae: Chilocorini) are redescribed, diagnosed, and illustrated. Lectotypes are designated for Endochilus compater Weise, Endochilus minor Weise, Endochilus plagiatus Sicard, Endochilus rubicundus Weise, and Endochilus styx Sicard. One new species is described: Endochilus abdominalissp. nov. Notes on the genus and nomenclatural history for each species are provided. A key for iden- tification of all species is presented. Adult characters concerning similarities of Endochilus to other genera of African Chilocorini are discussed. PMID:25373218

  3. Tumour detection and localization using 99Tcm-labelled OV-TL 3 Fab' in patients suspected of ovarian cancer

    International Nuclear Information System (INIS)

    Tibben, J.G.; Massuger, L.F.A.G.; Claessens, R.A.M.J.; Schijf, C.P.T.; Strijk, S.P.; Kenemans, P.; Corstens, F.H.M.; Pak, K.Y.

    1992-01-01

    Fab' fragments of the monoclonal antibody OV-TL 3, that recognizes an ovarian carcinoma-associated antigen (OA3), were labelled with 99 Tc m using D-glucarate as a ligand. Twenty patients suspected of having primary or recurrent ovarian cancer received intravenously 1 mg of the Fab' labelled with 740 MBq 99 Tc m . Both planar and single photon emission computed tomographic (SPECT) scintigraphy were performed up to 30 h after intravenous infusion. Imaging results were compared with X-ray computed tomography (CT), ultrasonography (US) and CA 125 serum level. Blood clearance was fast (t 1/2 of 9.5 h). Thirty-seven per cent of the injected dose (% ID) was excreted in the urine within the first 24 h, whereas 7% ID was excreted in the 24 h faeces. In one patient with an OA3 negative ovarian carcinoma, radioimmunoscintigraphy (RIS) did not visualize the tumour. In two other patients a benign ovarian cyst was found, also showing no elevated uptake. In 13 out of 17 patients ovarian cancer lesions were detected with RIS, whereas CT and US detected lesions in, respectively, 15 and 12 patients. Of 36 surgically defined tumour deposits larger than 1 cm in diameter, 53% were detected and localized with RIS, whereas CT and US detected 61 and 40%, respectively. Radioimmunoscintigraphy with 99 Tc m -OV-TL 3 Fab' is less distressing for the patients but the overall imaging performance is not improved when compared with 111 In-OV-TL 3 F(ab') 2 . (Author)

  4. The R117A variant of the Escherichia coli transacylase FabD synthesizes novel acyl-(acyl carrier proteins).

    Science.gov (United States)

    Marcella, Aaron M; Barb, Adam W

    2017-12-01

    The commercial impact of fermentation systems producing novel and biorenewable chemicals will flourish with the expansion of enzymes engineered to synthesize new molecules. Though a small degree of natural variability exists in fatty acid biosynthesis, the molecular space accessible through enzyme engineering is fundamentally limitless. Prokaryotic fatty acid biosynthesis enzymes build carbon chains on a functionalized acyl carrier protein (ACP) that provides solubility, stability, and a scaffold for interactions with the synthetic enzymes. Here, we identify the malonyl-coenzyme A (CoA)/holo-ACP transacylase (FabD) from Escherichia coli as a platform enzyme for engineering to diversify microbial fatty acid biosynthesis. The FabD R117A variant produced novel ACP-based primer and extender units for fatty acid biosynthesis. Unlike the wild-type enzyme that is highly specific for malonyl-CoA to produce malonyl-ACP, the R117A variant synthesized acetyl-ACP, succinyl-ACP, isobutyryl-ACP, 2-butenoyl-ACP, and β-hydroxybutyryl-ACP among others from holo-ACP and the corresponding acyl-CoAs with specific activities from 3.7 to 120 nmol min -1  mg -1 . FabD R117A maintained K M values for holo-ACP (~ 40 μM) and displayed small changes in K M for acetoacetyl-CoA (110 ± 30 μM) and acetyl-CoA (200 ± 70 μM) when compared to malonyl-CoA (80 ± 30 μM). FabD R117A represents a novel catalyst that synthesizes a broad range of acyl-acyl-ACPs.

  5. Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions.

    Science.gov (United States)

    Gao, Jian; Sasse, Joelle; Lewald, Kyle M; Zhalnina, Kateryna; Cornmesser, Lloyd T; Duncombe, Todd A; Yoshikuni, Yasuo; Vogel, John P; Firestone, Mary K; Northen, Trent R

    2018-04-10

    Beneficial plant-microbe interactions offer a sustainable biological solution with the potential to boost low-input food and bioenergy production. A better mechanistic understanding of these complex plant-microbe interactions will be crucial to improving plant production as well as performing basic ecological studies investigating plant-soil-microbe interactions. Here, a detailed description for ecosystem fabrication is presented, using widely available 3D printing technologies, to create controlled laboratory habitats (EcoFABs) for mechanistic studies of plant-microbe interactions within specific environmental conditions. Two sizes of EcoFABs are described that are suited for the investigation of microbial interactions with various plant species, including Arabidopsis thaliana, Brachypodium distachyon, and Panicum virgatum. These flow-through devices allow for controlled manipulation and sampling of root microbiomes, root chemistry as well as imaging of root morphology and microbial localization. This protocol includes the details for maintaining sterile conditions inside EcoFABs and mounting independent LED light systems onto EcoFABs. Detailed methods for addition of different forms of media, including soils, sand, and liquid growth media coupled to the characterization of these systems using imaging and metabolomics are described. Together, these systems enable dynamic and detailed investigation of plant and plant-microbial consortia including the manipulation of microbiome composition (including mutants), the monitoring of plant growth, root morphology, exudate composition, and microbial localization under controlled environmental conditions. We anticipate that these detailed protocols will serve as an important starting point for other researchers, ideally helping create standardized experimental systems for investigating plant-microbe interactions.

  6. Technetium-99 labelling of DD-3B6/22 antifibrin monoclonal antibody fragmented Fab' for thrombus imaging

    International Nuclear Information System (INIS)

    Lee, F-T.; Boniface, G.R.; Lambrecht, R.M.; Rylatt, D.B.; Bundesen, P.G.

    1993-01-01

    The antifibrin DD-3B6/22 monoclonal antibody Fab' fragment, a murine immunoglobulin, IgG3, has been labelled with technetium-99m ( 99mTc ) via a transchelation reaction, to specific activity in excess of 30 mCi/mg protein. The radiolabelling of Fab' was dependent on time, temperature, pH, antibody concentrations and nature intermediary transchelation complex used. The resultant radioconjugate was stable in vitro and in vivo. Blood clearance of 99m Tc-Fab' in rat followed two compartment kinetics with the half time of the fast phase being 0.5 h. The main route of excretion was via the kidneys with little uptake indicated by other tissues. The results suggest that the inherent specificity of the antibody, small molecular size, rapid plasma clearance, high specific radioactivity, together with the physical properties of the 99m Tc label, combine to make this labelled monoclonal antibody (MoAb), potentially suitable as a radiopharmaceutical for the scintigraphic detection of thrombi in humans. 17 refs., 3 tabs., 5 figs

  7. Dual Constant Domain-Fab: A novel strategy to improve half-life and potency of a Met therapeutic antibody.

    Science.gov (United States)

    Cignetto, Simona; Modica, Chiara; Chiriaco, Cristina; Fontani, Lara; Milla, Paola; Michieli, Paolo; Comoglio, Paolo M; Vigna, Elisa

    2016-06-01

    The kinase receptor encoded by the Met oncogene is a sensible target for cancer therapy. The chimeric monovalent Fab fragment of the DN30 monoclonal antibody (MvDN30) has an odd mechanism of action, based on cell surface removal of Met via activation of specific plasma membrane proteases. However, the short half-life of the Fab, due to its low molecular weight, is a severe limitation for the deployment in therapy. This issue was addressed by increasing the Fab molecular weight above the glomerular filtration threshold through the duplication of the constant domains, in tandem (DCD-1) or reciprocally swapped (DCD-2). The two newly engineered molecules showed biochemical properties comparable to the original MvDN30 in vitro, acting as full Met antagonists, impairing Met phosphorylation and activation of downstream signaling pathways. As a consequence, Met-mediated biological responses were inhibited, including anchorage-dependent and -independent cell growth. In vivo DCD-1 and DCD-2 showed a pharmacokinetic profile significantly improved over the original MvDN30, doubling the circulating half-life and reducing the clearance. In pre-clinical models of cancer, generated by injection of tumor cells or implant of patient-derived samples, systemic administration of the engineered molecules inhibited the growth of Met-addicted tumors. Copyright © 2016 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  8. A fully synthetic human Fab antibody library based on fixed VH/VL framework pairings with favorable biophysical properties

    Science.gov (United States)

    Tiller, Thomas; Schuster, Ingrid; Deppe, Dorothée; Siegers, Katja; Strohner, Ralf; Herrmann, Tanja; Berenguer, Marion; Poujol, Dominique; Stehle, Jennifer; Stark, Yvonne; Heßling, Martin; Daubert, Daniela; Felderer, Karin; Kaden, Stefan; Kölln, Johanna; Enzelberger, Markus; Urlinger, Stefanie

    2013-01-01

    This report describes the design, generation and testing of Ylanthia, a fully synthetic human Fab antibody library with 1.3E+11 clones. Ylanthia comprises 36 fixed immunoglobulin (Ig) variable heavy (VH)/variable light (VL) chain pairs, which cover a broad range of canonical complementarity-determining region (CDR) structures. The variable Ig heavy and Ig light (VH/VL) chain pairs were selected for biophysical characteristics favorable to manufacturing and development. The selection process included multiple parameters, e.g., assessment of protein expression yield, thermal stability and aggregation propensity in fragment antigen binding (Fab) and IgG1 formats, and relative Fab display rate on phage. The framework regions are fixed and the diversified CDRs were designed based on a systematic analysis of a large set of rearranged human antibody sequences. Care was taken to minimize the occurrence of potential posttranslational modification sites within the CDRs. Phage selection was performed against various antigens and unique antibodies with excellent biophysical properties were isolated. Our results confirm that quality can be built into an antibody library by prudent selection of unmodified, fully human VH/VL pairs as scaffolds. PMID:23571156

  9. Reclassification of leukemia among A-bomb survivors in Nagasaki using French-American-British (FAB) classification for acute leukemia

    International Nuclear Information System (INIS)

    Matsuo, Tatsuki; Tomonaga, Masao; Bennett, J.M.

    1988-01-01

    The concordance rate for diagnoses of atomic bomb-related cases of leukemia in Nagasaki was determined using the French-American-British (FAB) classification for acute leukemias and myelodysplastic syndromes (MDS). Two Radiation Effects Research Foundation (RERF) hematologists and one of the members (JMB) of the FAB cooperative group reviewed independently the peripheral blood and/or bone marrow smears from 193 people with leukemia or a related disorder. There was 85 % agreement in the identification of types and subtypes of acute leukemia. There was almost complete agreement for the diagnoses of non-FAB disorders (chronic myeloid leukemia (CML), adult T-cell leukemia (ATL) and others) resulting in overall concordance of 88.2 %. The present study suggest that the previously established leukemia types for about a quarter of the cases of acute leukemia and related disorders except CML should be changed. Considerable numbers of cases of ATL and MDS were involved in this series. The frequency of the former disease was not high in the high-dose irradiated group, but that of the latter was considerably high. All subtypes of AML except M3 and M6 were present in the high-dose group. The striking difference in CML incidence between Nagasaki and Hiroshima may continue to be a problem in relation to biological response to radiation exposure. (author)

  10. Reclassification of leukemia among A-bomb survivors in Nagasaki using French-American-British (FAB) classification for acute leukemia

    Energy Technology Data Exchange (ETDEWEB)

    Matsuo, Tatsuki; Tomonaga, Masao; Bennett, J.M. and others

    1988-06-01

    The concordance rate for diagnoses of atomic bomb-related cases of leukemia in Nagasaki was determined using the French-American-British (FAB) classification for acute leukemias and myelodysplastic syndromes (MDS). Two Radiation Effects Research Foundation (RERF) hematologists and one of the members (JMB) of the FAB cooperative group reviewed independently the peripheral blood and/or bone marrow smears from 193 people with leukemia or a related disorder. There was 85 % agreement in the identification of types and subtypes of acute leukemia. There was almost complete agreement for the diagnoses of non-FAB disorders (chronic myeloid leukemia (CML), adult T-cell leukemia (ATL) and others) resulting in overall concordance of 88.2 %. The present study suggest that the previously established leukemia types for about a quarter of the cases of acute leukemia and related disorders except CML should be changed. Considerable numbers of cases of ATL and MDS were involved in this series. The frequency of the former disease was not high in the high-dose irradiated group, but that of the latter was considerably high. All subtypes of AML except M3 and M6 were present in the high-dose group. The striking difference in CML incidence between Nagasaki and Hiroshima may continue to be a problem in relation to biological response to radiation exposure.

  11. The Response Regulator YycF Inhibits Expression of the Fatty Acid Biosynthesis Repressor FabT in Streptococcus pneumoniae

    Science.gov (United States)

    Mohedano, Maria L.; Amblar, Mónica; de la Fuente, Alicia; Wells, Jerry M.; López, Paloma

    2016-01-01

    The YycFG (also known as WalRK, VicRK, MicAB, or TCS02) two-component system (TCS) is highly conserved among Gram-positive bacteria with a low G+C content. In Streptococcus pneumoniae the YycF response regulator has been reported to be essential due to its control of pcsB gene expression. Previously we showed that overexpression of yycF in S. pneumoniae TIGR4 altered the transcription of genes involved in cell wall metabolism and fatty acid biosynthesis, giving rise to anomalous cell division and increased chain length of membrane fatty acids. Here, we have overexpressed the yycFG system in TIGR4 wild-type strain and yycF in a TIGR4 mutant depleted of YycG, and analyzed their effects on expression of proteins involved in fatty acid biosynthesis during activation of the TCS. We demonstrate that transcription of the fab genes and levels of their products were only altered in the YycF overexpressing strain, indicating that the unphosphorylated form of YycF is involved in the regulation of fatty acid biosynthesis. In addition, DNA-binding assays and in vitro transcription experiments with purified YycF and the promoter region of the FabTH-acp operon support a direct inhibition of transcription of the FabT repressor by YycF, thus confirming the role of the unphosphorylated form in transcriptional regulation. PMID:27610104

  12. Activation of Exogenous Fatty Acids to Acyl-Acyl Carrier Protein Cannot Bypass FabI Inhibition in Neisseria*

    Science.gov (United States)

    Yao, Jiangwei; Bruhn, David F.; Frank, Matthew W.; Lee, Richard E.; Rock, Charles O.

    2016-01-01

    Neisseria is a Gram-negative pathogen with phospholipids composed of straight chain saturated and monounsaturated fatty acids, the ability to incorporate exogenous fatty acids, and lipopolysaccharides that are not essential. The FabI inhibitor, AFN-1252, was deployed as a chemical biology tool to determine whether Neisseria can bypass the inhibition of fatty acid synthesis by incorporating exogenous fatty acids. Neisseria encodes a functional FabI that was potently inhibited by AFN-1252. AFN-1252 caused a dose-dependent inhibition of fatty acid synthesis in growing Neisseria, a delayed inhibition of growth phenotype, and minimal inhibition of DNA, RNA, and protein synthesis, showing that its mode of action is through inhibiting fatty acid synthesis. Isotopic fatty acid labeling experiments showed that Neisseria encodes the ability to incorporate exogenous fatty acids into its phospholipids by an acyl-acyl carrier protein-dependent pathway. However, AFN-1252 remained an effective antibacterial when Neisseria were supplemented with exogenous fatty acids. These results demonstrate that extracellular fatty acids are activated by an acyl-acyl carrier protein synthetase (AasN) and validate type II fatty acid synthesis (FabI) as a therapeutic target against Neisseria. PMID:26567338

  13. Fab-based inhibitors reveal ubiquitin independent functions for HIV Vif neutralization of APOBEC3 restriction factors.

    Directory of Open Access Journals (Sweden)

    Jennifer M Binning

    2018-01-01

    Full Text Available The lentiviral protein Viral Infectivity Factor (Vif counteracts the antiviral effects of host APOBEC3 (A3 proteins and contributes to persistent HIV infection. Vif targets A3 restriction factors for ubiquitination and proteasomal degradation by recruiting them to a multi-protein ubiquitin E3 ligase complex. Here, we describe a degradation-independent mechanism of Vif-mediated antagonism that was revealed through detailed structure-function studies of antibody antigen-binding fragments (Fabs to the Vif complex. Two Fabs were found to inhibit Vif-mediated A3 neutralization through distinct mechanisms: shielding A3 from ubiquitin transfer and blocking Vif E3 assembly. Combined biochemical, cell biological and structural studies reveal that disruption of Vif E3 assembly inhibited A3 ubiquitination but was not sufficient to restore its packaging into viral particles and antiviral activity. These observations establish that Vif can neutralize A3 family members in a degradation-independent manner. Additionally, this work highlights the potential of Fabs as functional probes, and illuminates how Vif uses a multi-pronged approach involving both degradation dependent and independent mechanisms to suppress A3 innate immunity.

  14. Distribuição e alimentação de duas espécies simpátricas de piranhas Serrasalmus maculatus e Pygocentrus nattereri (Characidae, Serrasalminae do rio Ibicuí, Rio Grande do Sul, Brasil Distribution and feeding of two sympatric species of piranhas Serrasalmus maculatus and Pygocentrus nattereri (Characidae, Serrasalminae of the Ibicuí river, State of Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Everton R. Behr

    2008-12-01

    Full Text Available Duas espécies de piranhas ocorrem no rio Ibicuí, um dos principais afluentes do rio Uruguai. Com o objetivo de analisar a distribuição e a alimentação destas espécies, foram realizadas coletas de dezembro de 1999 a janeiro de 2002, utilizando-se diferentes artes de pesca em três pontos do rio, sendo que cada ponto foi amostrado em dois ambientes (lêntico e lótico. Foram capturados 203 indivíduos de Serrasalmus maculatus Kner, 1858 e 86 de Pygocentrus nattereri Kner, 1858, sendo que a maior captura de ambas as espécies ocorreu no ambiente lêntico e próximo ao rio Uruguai, onde P. nattereri parece estar mais concentrada. O número de exemplares capturados por horário não apresentou variações significativas em P. nattereri enquanto que S. maculatus apresentou a maior captura na revisão da meia-noite. Juvenis de S. maculatus (2-4 cm de comprimento padrão consumiram preferencialmente nadadeiras e insetos. Nas demais classes de tamanho de ambas as espécies, restos de peixes foi o alimento predominante, havendo sobreposição alimentar intra e interespecífica nas maiores classes (8-16 cm e >16 cm de comprimento padrão. Não foram constatadas diferenças no índice de repleção entre os horários e os ambientes para as duas espécies.Two species of piranhas occur in the Ibicuí River, one of the main tributaries of the Uruguay River in southern Brazil, Serrasalmus maculatus Kner, 1858 and Pygocentrus nattereri Kner, 1858. In order to analyze the distribution and feeding habits of these species we conducted collecting expeditions from December 1999 to January 2002, using a variety of fishing gears in three sites along the river, both in lentic and lotic habitats. A total of 203 individuals of S. maculatus and 86 of P. nattereri were caught, mostly in lentic habitats and closer to the Uruguay River, were P. nattereri seems to be more concentrated. The number of individuals captured at different times did not show variation in P

  15. Rove beetles of medical importance in Brazil (Coleoptera, Staphylinidae, Paederinae

    Directory of Open Access Journals (Sweden)

    Juliana S. Vieira

    2014-09-01

    Full Text Available Rove beetles of medical importance in Brazil (Coleoptera, Staphylinidae, Paederinae. The rove beetles of the genus Paederus Fabricius, 1775 are the most important group within Coleoptera causing dermatitis around the world. The medical importance of Paederus depends on its toxic hemolymph released when these beetles are crushed on human skin. The effects are mainly dermatitis linearis and some sporadic cases of conjunctivitis. In Brazil seven species of Paederus are known to cause dermatitis: P. amazonicus Sharp, 1876, P. brasiliensis Erichson, 1840, P. columbinus Laporte, 1835, P. ferus Erichson, 1840, P. mutans Sharp, 1876, P. protensus Sharp, 1876 stat. rev., and Paederus rutilicornis Erichson, 1840. Paederus mutans and P. protensus are for the first time recorded as of medical importance, whereas the record of P. rutilicornis in Brazil is doubtful. All seven species are redescribed and a dichotomous key is provided. The geographic distributions of all species are documented. The results provided here include the most recent and relevant taxonomic revision of Paederus of the Neotropical region, the first identification key for Brazilian species and the increase of recorded species of medical importance in the world.

  16. A kit to prepare {sup 111}In-DTPA-trastuzumab (Herceptin) Fab fragments injection under GMP conditions for imaging or radioimmunoguided surgery of HER2-positive breast cancer

    Energy Technology Data Exchange (ETDEWEB)

    Scollard, Deborah A.; Chan, Conrad [Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, M5S 3M2 (Canada); Holloway, Claire M.B. [Department of Surgery, Sunnybrook Health Sciences Centre, Toronto, ON, M4N 1H1 (Canada); Reilly, Raymond M., E-mail: raymond.reilly@utoronto.c [Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, M5S 3M2 (Canada); Department of Medical Imaging, University of Toronto, Toronto, ON, M5S 3E2 (Canada); Toronto General Research Institute, University Health Network, Toronto, ON, M5G 2M9 (Canada)

    2011-01-15

    Introduction: The human epidermal growth factor receptor-2 (HER2) gene is amplified in 25% of invasive breast cancers, and receptor overexpression has been noted in up to 60% of early stages of the disease [ductal carcinoma in situ (DCIS)]. Preclinical studies have revealed high tumor/blood ratios (>27:1) for {sup 111}In-labeled Fab fragments of the HER2 monoclonal antibody, trastuzumab (Herceptin) ({sup 111}In-DTPA-trastuzumab Fab) at 72 h pi in athymic mice bearing subcutaneous human breast cancer xenografts. Our aim in this study was to formulate a kit for preparation of {sup 111}In-DTPA-trastuzumab Fab injection under good manufacturing practice (GMP) conditions suitable for human administration in a Phase I clinical trial of imaging and radioimmunoguided surgery (RIGS) of HER2-positive breast cancer. Methods: Fab fragments were produced by digestion of trastuzumab IgG (Herceptin) with immobilized papain for 20 h at 37{sup o}C. Fab fragments were purified by ultrafiltration, then reacted with a 10-fold molar excess of diethylenetriaminepentaacetic acid (DTPA) dianhydride. DTPA-Fab fragments were purified, then sterilized by filtration into unit dose glass vials (kits). Kits were tested against specifications for volume (0.9-1.1 ml), protein concentration (0.45-0.55 mg/ml), pH (5.5-6.5), DTPA substitution (0.5-4.0 mol DTPA/mol Fab), appearance (clear, colorless and particle free), labeling efficiency ({>=}85%), and sterility and apyrogenicity (USP XXXII). Immunoreactivity of {sup 111}In-DTPA-trastuzumab Fab towards HER2 was measured by saturation radioligand binding assays using SKBR-3 human breast cancer cells (specifications: K{sub a}=0.6-9.6x10{sup 7} L/mol; B{sub max}=0.6-10.4x10{sup 6} sites/cell). {sup 111}In-DTPA-trastuzumab Fab injection was prepared by adding 80-100 MBq of {sup 111}InCl{sub 3} to a single kit vial and incubating for 30 min at room temperature. {sup 111}In-DTPA-trastuzumab Fab was assayed for the amount of radioactivity and tested for p

  17. Conjugation of 10 kDa Linear PEG onto Trastuzumab Fab' Is Sufficient to Significantly Enhance Lymphatic Exposure while Preserving in Vitro Biological Activity.

    Science.gov (United States)

    Chan, Linda J; Ascher, David B; Yadav, Rajbharan; Bulitta, Jürgen B; Williams, Charlotte C; Porter, Christopher J H; Landersdorfer, Cornelia B; Kaminskas, Lisa M

    2016-04-04

    The lymphatic system is a major conduit by which many diseases spread and proliferate. There is therefore increasing interest in promoting better lymphatic drug targeting. Further, antibody fragments such as Fabs have several advantages over full length monoclonal antibodies but are subject to rapid plasma clearance, which can limit the lymphatic exposure and activity of Fabs against lymph-resident diseases. This study therefore explored ideal PEGylation strategies to maximize biological activity and lymphatic exposure using trastuzumab Fab' as a model. Specifically, the Fab' was conjugated with single linear 10 or 40 kDa PEG chains at the hinge region. PEGylation led to a 3-4-fold reduction in binding affinity to HER2, but antiproliferative activity against HER2-expressing BT474 cells was preserved. Lymphatic pharmacokinetics were then examined in thoracic lymph duct cannulated rats after intravenous and subcutaneous dosing at 2 mg/kg, and the data were evaluated via population pharmacokinetic modeling. The Fab' displayed limited lymphatic exposure, but conjugation of 10 kDa PEG improved exposure by approximately 11- and 5-fold after intravenous (15% dose collected in thoracic lymph over 30 h) and subcutaneous (9%) administration, respectively. Increasing the molecular weight of the PEG to 40 kDa, however, had no significant impact on lymphatic exposure after intravenous (14%) administration and only doubled lymphatic exposure after subcutaneous administration (18%) when compared to 10 kDa PEG-Fab'. The data therefore suggests that minimal PEGylation has the potential to enhance the exposure and activity of Fab's against lymph-resident diseases, while no significant benefit is achieved with very large PEGs.

  18. Targeting Mast Cells and Basophils with Anti-FcεRIα Fab-Conjugated Celastrol-Loaded Micelles Suppresses Allergic Inflammation.

    Science.gov (United States)

    Peng, Xia; Wang, Juan; Li, Xianyang; Lin, Lihui; Xie, Guogang; Cui, Zelin; Li, Jia; Wang, Yuping; Li, Li

    2015-12-01

    Mast cells and basophils are effector cells in the pathophysiology of allergic diseases. Targeted elimination of these cells may be a promising strategy for the treatment of allergic disorders. Our present study aims at targeted delivery of anti-FcεRIα Fab-conjugated celastrol-loaded micelles toward FcεRIα receptors expressed on mast cells and basophils to have enhanced anti-allergic effect. To achieve this aim, we prepared celastrol-loaded (PEO-block-PPO-block-PEO, Pluronic) polymeric nanomicelles using thin-film hydration method. The anti-FcεRIα Fab Fragment was then conjugated to carboxyl groups on drug-loaded micelles via EDC amidation reaction. The anti-FcεRIα Fab-conjugated celastrol-loaded micelles revealed uniform particle size (93.43 ± 12.93 nm) with high loading percentage (21.2 ± 1.5% w/w). The image of micelles showed oval and rod like. The anti-FcεRIα Fab-conjugated micelles demonstrated enhanced cellular uptake and cytotoxity toward target KU812 cells than non-conjugated micelles in vitro. Furthermore, diffusion of the drug into the cells allowed an efficient induction of cell apoptosis. In mouse model of allergic asthma, treatment with anti-FcεRIα Fab-conjugated micelles increased lung accumulation of micelles, and significantly reduced OVA-sIgE, histamine and Th2 cytokines (IL-4, IL-5, TNF-α) levels, eosinophils infiltration and mucus production. In addition, in mouse model of passive cutaneous anaphylaxis, anti-FcεRIα Fab-conjugated celastrol-loaded micelles treatment significantly decreased extravasated evan's in the ear. These results indicate that anti-FcεRIα Fab-conjugated celastrol-loaded micelles can target and selectively kill mast cells and basophils which express FcεRIα, and may be efficient reagents for the treatment of allergic disorders and mast cell related diseases.

  19. Cytosolic expression of functional Fab fragments in Escherichia coli using a novel combination of dual SUMO expression cassette and EnBase® cultivation mode.

    Science.gov (United States)

    Rezaie, F; Davami, F; Mansouri, K; Agha Amiri, S; Fazel, R; Mahdian, R; Davoudi, N; Enayati, S; Azizi, M; Khalaj, V

    2017-05-08

    The Escherichia coli expression system is highly effective in producing recombinant proteins. However, there are some limitations in this system, especially in obtaining correctly folded forms of some complex proteins such as Fab fragments. To improve the solubility and folding quality of Fab fragments, we have examined the effect of simultaneous application of a SUMO fusion tag, EnBase ® cultivation mode and a redox mutant strain in the E. coli expression system. A bicistronic gene construct was designed to express an antivascular endothelial growth factor (VEGF) Fab fragment as a model system. The construct contained a dual SUMO fusion gene fragment to encode SUMO-tagged heavy and light chains. While the expression of the construct in batch cultures of BL21 or SHuffle ® transformants produced insoluble and unfolded products, the induction of the transformants in EnBase ® medium resulted in soluble and correctly folded Fab fragment, reaching as high as 19% of the total protein in shuffle strain. The functional assays indicated that the biological activity of the target Fab is similar to the commercial anti-VEGF, Lucentis ® . This study demonstrated that the combination of SUMO fusion technology, EnBase ® cultivation system and recruiting a redox mutant of E. coli can efficiently enhance the solubility and productivity of recombinant Fab fragments. The presented strategy provides not only a novel method to produce soluble and active form of an anti-VEGF Fab but also may use in the efficient production of other antibody fragments. © 2017 The Society for Applied Microbiology.

  20. California State University, Bakersfield Fab Lab: "Making" A Difference in Middle School Students' STEM Attitudes

    Science.gov (United States)

    Medina, Andrea Lee

    The digital fabrication lab, or Fab Lab, at California State University, Bakersfield provided a 1-week, half-day summer program for local area middle school students. The purpose of this study was to examine the effect this summer program had on their attitudes towards math and science. The theoretical framework used for this study was based on Papert’s (1980) theory of constructionism and Bandura’s (1977) self-efficacy theory. Papert’s interest in how learners engaged in discussions with the items they made, and how these interactions increased self-guided learning, promoted the development of new knowledge. Self-efficacy, or one’s belief in his or her ability to perform behaviors necessary to produce specific achievements, increases as a result of the self-guided learning. These beliefs are proposed to influence future aspirations and the commitment to them. Results of the paired t-tests show a marked difference between 2016 participants (n= 49) and 2017 participants (n=31). Of the 2016 participants, no overall significance was found on attitudes towards math or science, but male attitudes within the math subset did show significance. The results of the 2017 program do show statistical significance in the area of science for females. It is hypothesized that the difference in results were due to the delivery of the program between the 2 years. Further research is necessary to confirm this hypothesis.

  1. Fabrication of Quench Condensed Thin Films Using an Integrated MEMS Fab on a Chip

    Science.gov (United States)

    Lally, Richard; Reeves, Jeremy; Stark, Thomas; Barrett, Lawrence; Bishop, David

    Atomic calligraphy is a microelectromechanical systems (MEMS)-based dynamic stencil nanolithography technique. Integrating MEMS devices into a bonded stacked array of three die provides a unique platform for conducting quench condensed thin film mesoscopic experiments. The atomic calligraphy Fab on a Chip process incorporates metal film sources, electrostatic comb driven stencil plate, mass sensor, temperature sensor, and target surface into one multi-die assembly. Three separate die are created using the PolyMUMPs process and are flip-chip bonded together. A die containing joule heated sources must be prepared with metal for evaporation prior to assembly. A backside etch of the middle/central die exposes the moveable stencil plate allowing the flux to pass through the stencil from the source die to the target die. The chip assembly is mounted in a cryogenic system at ultra-high vacuum for depositing extremely thin films down to single layers of atoms across targeted electrodes. Experiments such as the effect of thin film alloys or added impurities on their superconductivity can be measured in situ with this process.

  2. Clinical experience of the radioimmunoscintigraphy with the 123-I-anti-CEA-Fab(No. 35)-fragment

    International Nuclear Information System (INIS)

    Meili, A.; Bekier, A.; Schulthess, G.K. von; Mach, J.P.

    1986-01-01

    Purified monoclonal murine 123-I-Fab-anti-CEA(Nr.35) were applied to 7 patients in a prospective study. Compared with the previous tested intact 131-I-labelled antibody (Nr. 202) the new preparation had many advantages for imaging: 2.7 times more counting efficiency of the gamma camera, higher activity utilizable and better accumulation in the tumor with sufficient image contrast in the ECT. Fifteen until sixteen hours after administration of 20 mCi 123-I the countrate in the pelvis was 130000/20 sec. In all patients the primary tumor was visualized, two patients demonstrated regional lymph node involvement and one patient liver metastasis in the left lobe. All findings were confirmed by surgery and histopathology. The average tumor/mucosa ratio was 3.5:1. The activity in the tumor 15 h after application was 0.015 per cent/g tumor/mCi 123-I used. 4/7 patients with colorectal cancer over 3.5 cm in diameter demonstrated normal plasma values of CEA < 3 microg/l. One patient with rising CEA plasma level was suspected of relapse, the focal accumulation corresponded with a mass lesion in the right sacrum in CT-scan. After the preliminary results the further applicability is recognizable: staging and screening before and after therapy, handicapped by high costs at time. A good advantage is the possibility of specific and organ independent diagnosis of tumor, which contents the corresponding surface antigen. (author)

  3. Clinical utility of immunoscintigraphy with the Immu-4 99mTc-Fab' antibody

    International Nuclear Information System (INIS)

    Moffat, F.L.; Pinsky, C.M.; Hammershaimb, L.; Petrelli, N.J.; Patt, Y.Z.; Whaley, F.S.; Goldenberg, D.M.

    1996-01-01

    The purpose of these prospective, pivotal studies was to assess the performance and the potential clinical impact of a new antibody imaging agent, CEA-Scan TM , in 210 pre-surgical patients with advanced recurrent or metastatic colorectal carcinomas. CEA-Scan TM , an anti-carcinoembryonic antigen (CEA) Fab' antibody fragment labeled with 99m Tc, was injected IV and external scintigraphy performed 2-5 h and 18-24 h later. Imaging with conventional diagnostic modalities (CDM) was also performed, and findings were confirmed by surgery and histology. The sensitivity of CEA-Scan TM was superior to that of CDM in the extrahepatic abdomen (55 % vs 32 %, ρ 0,007) and pelvis (69 % vs 48 %, p = 0,005), and CEA-Scan TM findings complemented those of CDM in the liver. Among 122 patients with known disease, the positive predictive value was significantly higher when both modalities were positive (98 %) as compared to each alone (68 % - 70 %), potentially obviating the need for histological confirmation when both tests are positive. Imaging accuracy also was significantly improved y adding CEA-Scan TM to CDM. In 88 patients with occult cancer, imaging accuracy was enhanced significantly by CEA-Scan TM combined with CDM (61 % vs 33 %). Potential clinical benefit from CEA-Scan TM was demonstrated in 89 of 210 patients. Only two patients developed human anti-mouse antibodies (HAMA) to CEA-Scan TM after a single injection, and none of 19 assessable patients after two injections. (author)

  4. Distribution and dosimetry of indium-111 labeled F(ab')2 fragments in humans

    International Nuclear Information System (INIS)

    Doherty, P.; Schwinger, R.; King, M.; Gionet, M.

    1986-01-01

    The purpose of this study was to obtain quantitative biodistribution data in patients injected with the indium-111 labeled F(ab') 2 fragments of mouse monoclonal antibody. From this data dosimetric calculations were made for the individual organs. The authors also evaluated the quantitative properties of SPECT in this application and compared it with the more conventional two view planar technique in both phantom and patient studies. For one antibody (19-9) the mean dose in rads/mCi for the organs of highest accumulation, namely, the liver and kidneys was 3.2 and 2.6 respectively. Preliminary data from another antibody (OC 125) showed much higher blood levels and a significantly lower liver dose of 2.3 indicating that antibody type is another significant determinant in dosimetry. The SPECT approach particularly in the presence of background activity, was more accurate in the phantom studies and resulted in larger estimated doses in the patient studies. Also, SPECT has the added advantage of providing an index of organ volume, which has to be balanced with the fact the planar is more rapid, and does not require special hardware. 24 references, 5 figures, 1 table

  5. A new gonad-infecting species of Philometra (Nematoda: Philometridae) from the Atlantic Spanish mackerel Scomberomorus maculatus (Scombridae) off the Atlantic Coast of Florida and South Carolina.

    Science.gov (United States)

    Moravec, František; Bakenhaster, Micah; de Buron, Isaure

    2013-04-01

    A new nematode species, Philometra atlantica n. sp. (Philometridae), is described from male and female specimens found in the ovary of the Atlantic Spanish mackerel, Scomberomorus maculatus (Mitchill) (Scombridae, Perciformes), off the Atlantic coast of Florida and South Carolina. Based on light and scanning electron microscopy examination, the new species differs from most other gonad-infecting Philometra spp. in the length of spicules (111-126 μm), number and arrangement of genital papillae, and a U-shaped, dorsally interrupted caudal mound in the male. A unique feature among all gonad-infecting philometrids is the presence of 2 reflexed dorsal barbs on the distal end of the gubernaculum. From a few congeneric, gonad-infecting species with unknown males, it can be distinguished by some morphological and biometrical features found in gravid females (body length, length of first-stage larvae or esophagus, structure of caudal end) and by the host type (fish family) and geographical distribution. Philometra atlantica is the fourth valid gonad-infecting species of Philometra reported from fishes of the family Scombridae.

  6. Helmintos parasitos das espécies Scomberomorus cavalla (Cuvier e Scomberomorus maculatus (Mitchill do litoral cearense: Contracaecum fortalezae sp. n. (Nematoda, Ascaridoidea

    Directory of Open Access Journals (Sweden)

    Vera Lucia M. Klein

    1973-01-01

    Full Text Available De nematódeos encontrados parasitando estômago, intestinos delgado e grosso de Scomberomorus cavalla (Cuv. e Scomberomorus maculatus (Mitch. o autor propões para o gênero Contracaecum Railliet et Henry, 1912 uma nova espécie, C. fortalezae sp. n., que mais se aproxima de C. clavatum (Rud., 1809 Baylis, 1920 dela se diferenciando por possuir espículos desiguais, um curto ovejetor, útero opistodelfo, ovos menores e por apresentar em todos exemplares estudados ceco intestinal curto e ceco esofagiano longo.In this paper the author proposes a new species of the genus Contracaecum Railliet et Henry, 1912. Of the species under Contracaecum, Contracaecum clavatum (Rudolphi, 1809 Baylis, 1920 is the nearest to Contracaecum fortalezae sp. n. differing from the latter by the following aspects: The males have unequal spicules. The females have a short ovijector and parallel uteri directed backward and the eggs are smaller. In both sexes the intestinal cecum is always short and the ventricular posterior appendix is always long, while Contracaecum clavatum presents these structures in an inverse way, considering the lenght of them. Besides, the thorn-like formations at the posterior end of the body are different.

  7. Catalogue of Tenebrionidae (Coleoptera of North America

    Directory of Open Access Journals (Sweden)

    Yves Bousquet

    2018-01-01

    Full Text Available This catalogue includes all valid family-group (8 subfamilies, 52 tribes, 14 subtribes, genus-group (349 genera, 86 subgenera, and species-group names (2825 species, 215 subspecies of darkling beetles (Coleoptera: Tenebrionidae known to occur in North America1 and their available synonyms. Data on extant, subfossil and fossil taxa are given. For each name the author and year and page number of the description are provided, with additional information (e.g., type species for genus-group names, author of synonymies for invalid taxa depending on the taxon rank. Several new nomenclatural acts are included. One new genus, Lepidocnemeplatia Bousquet and Bouchard, is described. Spelaebiosis Bousquet and Bouchard [for Ardoinia Özdikmen, 2004], Blapstinus marcuzzii Aalbu [for Blapstinus kulzeri Marcuzzi, 1977], and Hymenorus campbelli Bouchard [for Hymenorus oculatus Doyen and Poinar, 1994] are proposed as new replacement names. Supporting evidence is provided for the conservation of usage of Tarpela micans (Fabricius, 1798 nomen protectum over Tarpela vittata (Olivier, 1793 nomen oblitum. The generic names Psilomera Motschulsky, 1870 [= Stenomorpha Solier, 1836], Steneleodes Blaisdell, 1909 [= Xysta Eschscholtz, 1829], Ooconibius Casey, 1895 and Euconibius Casey, 1895 [= Conibius LeConte, 1851] are new synonyms (valid names in square brackets. The following 127 new synonymies of species-group names, listed in their original combination, are proposed (valid names, in their current combination, placed in square brackets: Bothrasida mucorea Wilke, 1922 [= Pelecyphorus guanajuatensis (Champion, 1884]; Parasida zacualpanicola Wilke, 1922 [= Pelecyphorus asidoides Solier, 1836]; Stenosides kulzeri Pallister, 1954, Stenosides bisinuatus Pallister, 1954, and Parasida trisinuata Pallister, 1954 [= Pelecyphorus dispar (Champion, 1892]; Asida favosa Champion, 1884 and Asida similata Champion, 1884 [= Pelecyphorus fallax (Champion, 1884]; Ologlyptus bicarinatus

  8. Imaging of HER2/neu-positive BT-474 human breast cancer xenografts in athymic mice using 111In-trastuzumab (Herceptin) Fab fragments

    International Nuclear Information System (INIS)

    Tang Ying; Wang, Judy; Scollard, Deborah A.; Mondal, Hridya; Holloway, Claire; Kahn, Harriette J.; Reilly, Raymond M.

    2005-01-01

    Trastuzumab (Herceptin) Fab were prepared by digestion of intact IgG with immobilized papain, derivatized with diethylenetriaminepentaacetic acid (DTPA) and radiolabeled with 111 In. The dissociation constant (K d ) for binding of Fab to HER2/neu-positive SK-BR-3 human breast cancer cells was two- to threefold higher than for intact IgG (14-36 vs. 8-14 nM). The binding affinity was not significantly decreased after DTPA derivatization (K d =47 nM). 111 In-trastuzumab Fab localized specifically in HER2/neu-positive BT-474 human breast cancer xenografts in athymic mice with tumor uptake of 7.8±0.7% injected dose (ID)/g and tumor/blood ratio of 25.2±1.6 at 72 h postinjection compared with 2.7±0.7% ID/g and 7.0±0.9 for 111 In-HuM195 anti-CD33 Fab (significantly different, P 111 In-trastuzumab Fab as early as 24 h postinjection

  9. Label-free Fab and Fc affinity/avidity profiling of the antibody complex half-life for polyclonal and monoclonal efficacy screening.

    Science.gov (United States)

    Read, Thomas; Olkhov, Rouslan V; Williamson, E Diane; Shaw, Andrew M

    2015-09-01

    A unified approach to affinity screening for Fab and Fc interactions of an antibody for its antigen and FcγR receptor has been developed. An antigen array is used for the Fab affinity and cross-reactivity screening and protein A/G proxy is the FcγR receptor. The affinities are derived using a simple 1:1 binding model with a consistent error analysis. The association and dissociation kinetics are measured over optimised times for accurate determination. The Fab/Fc affinities are derived for ten antibodies: mAb-actin (mouse), pAb-BSA (sheep), pAb-collagen V (rabbit), pAb-CRP (goat), mAb-F1 (mouse), mAbs (mouse) 7.3, 12.3, 29.3, 36.3 and 46.3 raised against LcrV in Yersinia pestis. The rate of the dissociation of antigen-antibody complexes relates directly to their immunological function as does the Fc-FcγR complex and a new half-life plot has been defined with a Fab/Fc half-life range of 17-470 min. The upper half-life value points to surface avidity. Two antibodies that are protective as an immunotherapy define a Fab half-life >250 min and an Fc half-life >50 min as characteristics of ideal interactions which can form the basis of an antibody screen for immunotherapy.

  10. Crystallization and preliminary X-ray diffraction analysis of prion protein bound to the Fab fragment of the POM1 antibody

    International Nuclear Information System (INIS)

    Baral, Pravas Kumar; Wieland, Barbara; Swayampakula, Mridula; Polymenidou, Magdalini; Aguzzi, Adriano; Kav, Nat N. V.; James, Michael N. G.

    2011-01-01

    The complex of MoPrP(120–232) and Fab POM1 has been crystallized (space group C2, unit-cell parameters a = 83.68, b = 106.9, c = 76.25 Å, β = 95.6°). Diffraction data to 2.30 Å resolution have been collected using synchrotron radiation. Prion diseases are neurodegenerative diseases that are characterized by the conversion of the cellular prion protein PrP c to the pathogenic isoform PrP sc . Several antibodies are known to interact with the cellular prion protein and to inhibit this transition. An antibody Fab fragment, Fab POM1, was produced that recognizes a structural motif of the C-terminal domain of mouse prion protein. To study the mechanism by which Fab POM1 recognizes and binds the prion molecule, the complex between Fab POM1 and the C-terminal domain of mouse prion (residues 120–232) was prepared and crystallized. Crystals of this binary complex belonged to the monoclinic space group C2, with unit-cell parameters a = 83.68, b = 106.9, c = 76.25 Å, β = 95.6°

  11. Structural and biophysical characterization of an epitope-specific engineered Fab fragment and complexation with membrane proteins: implications for co-crystallization.

    Science.gov (United States)

    Johnson, Jennifer L; Entzminger, Kevin C; Hyun, Jeongmin; Kalyoncu, Sibel; Heaner, David P; Morales, Ivan A; Sheppard, Aly; Gumbart, James C; Maynard, Jennifer A; Lieberman, Raquel L

    2015-04-01

    Crystallization chaperones are attracting increasing interest as a route to crystal growth and structure elucidation of difficult targets such as membrane proteins. While strategies to date have typically employed protein-specific chaperones, a peptide-specific chaperone to crystallize multiple cognate peptide epitope-containing client proteins is envisioned. This would eliminate the target-specific chaperone-production step and streamline the co-crystallization process. Previously, protein engineering and directed evolution were used to generate a single-chain variable (scFv) antibody fragment with affinity for the peptide sequence EYMPME (scFv/EE). This report details the conversion of scFv/EE to an anti-EE Fab format (Fab/EE) followed by its biophysical characterization. The addition of constant chains increased the overall stability and had a negligible impact on the antigen affinity. The 2.0 Å resolution crystal structure of Fab/EE reveals contacts with larger surface areas than those of scFv/EE. Surface plasmon resonance, an enzyme-linked immunosorbent assay, and size-exclusion chromatography were used to assess Fab/EE binding to EE-tagged soluble and membrane test proteins: namely, the β-barrel outer membrane protein intimin and α-helical A2a G protein-coupled receptor (A2aR). Molecular-dynamics simulation of the intimin constructs with and without Fab/EE provides insight into the energetic complexities of the co-crystallization approach.

  12. Specific Conjugation of the Hinge Region for Homogeneous Preparation of Antibody Fragment-Drug Conjugate: A Case Study for Doxorubicin-PEG-anti-CD20 Fab' Synthesis.

    Science.gov (United States)

    Zhou, Zhan; Zhang, Jing; Zhang, Yan; Ma, Guanghui; Su, Zhiguo

    2016-01-20

    Conventional preparation strategies for antibody-drug conjugates (ADCs) result in heterogeneous products with various molecular sizes and species. In this study, we developed a homogeneous preparation strategy by site-specific conjugation of the anticancer drug with an antibody fragment. The model drug doxorubicin (DOX) was coupled to the Fab' fragment of anti-CD20 IgG at its permissive sites through a heterotelechelic PEG linker, generating an antibody fragment-drug conjugate (AFDC). Anti-CD20 IgG was digested and reduced specifically with β-mercaptoethylamine to generate the Fab' fragment with two free mercapto groups in its hinge region. Meanwhile, DOX was conjugated with α-succinimidylsuccinate ω-maleimide polyethylene glycol (NHS-PEG-MAL) to form MAL-PEG-DOX, which was subsequently linked to the free mercapto containing Fab' fragment to form a Fab'-PEG-DOX conjugate. The dual site-specific bioconjugation was achieved through the combination of highly selective reduction of IgG and introduction of heterotelechelic PEG linker. The resulting AFDC provides an utterly homogeneous product, with a definite ratio of one fragment to two drugs. Laser confocal microscopy and cell ELISA revealed that the AFDC could accumulate in the antigen-positive Daudi tumor cell. In addition, the Fab'-PEG-DOX retained appreciable targeting ability and improved antitumor activity, demonstrating an excellent therapeutic effect on the lymphoma mice model for better cure rate and significantly reduced side effects.

  13. Generation and selection of naïve Fab library for parasitic antigen: Anti-BmSXP antibodies for lymphatic filariasis.

    Science.gov (United States)

    Omar, Noorsharmimi; Hamidon, Nurul Hamizah; Yunus, Muhammad Hafiznur; Noordin, Rahmah; Choong, Yee Siew; Lim, Theam Soon

    2018-05-01

    Phage display has been applied successfully as a tool for the generation of monoclonal antibodies (mAbs). Naive antibody libraries are unique as they are able to overcome several limitations associated with conventional mAb generation methods like the hybridoma technology. Here, we performed an in vitro selection and generation of Fab antibodies against Brugia malayi SXP protein (BmSXP), a recombinant antigen for the detection of lymphatic filariasis. We developed a naïve multi ethnic Fab antibody library with an estimated diversity of 2.99 × 10 9 . The antibody library was used to screen for mAbs against BmSXP recombinant antigen. Soluble monoclonal Fab antibodies against BmSXP were successfully isolated from the naïve library. The Fab antibodies obtained were expressed and analyzed to show its binding capability. The diversity obtained from a pool of donors from various ethnic groups allowed for a diverse antibody library to be generated. The mAbs obtained were also functional in soluble form, which makes it useful for further downstream applications. We believe that the Fab mAbs are valuable for further studies and could also contribute to improvements in the diagnosis of filariasis. © 2017 International Union of Biochemistry and Molecular Biology, Inc.

  14. Human/murine chimeric 81C6 F(ab')2 fragment: preclinical evaluation of a potential construct for the targeted radiotherapy of malignant glioma

    International Nuclear Information System (INIS)

    Boskovitz, Abraham; Akabani, Gamal H.; Pegram, Charles N.; Bigner, Darrell D.; Zalutsky, Michael R.

    2004-01-01

    We have obtained encouraging responses in recent Phase I studies evaluating 131 I-labeled human/murine chimeric 81C6 anti-tenascin monoclonal antibody (ch81C6) administered into surgically-created tumor resection cavities in brain tumor patients. However, because the blood clearance is slow, hematologic toxicity has been higher than seen with murine 81C6 (mu81C6). In the current study, a series of paired-label experiments were performed in athymic mice bearing subcutaneous D-245 MG human glioma xenografts to compare the biodistribution of the fragment ch81C6 F(ab') 2 labeled using Iodogen to a) intact ch81C6, b) mu81C6, and c) ch81C6 F(ab') 2 labeled using N-succinimidyl 3-[ 131 I]iodobenzoate. Tumor retention of radioiodine activity for the F(ab') 2 fragment was comparable to that for intact ch81C6 for the first 24 h and to that for mu81C6 for the first 48 h; as expected, blood and other normal tissue levels declined faster for ch81C6 F(ab') 2. Radiation dosimetry calculations suggest that 131 I-labeled ch81C6 F(ab') 2 may warrant further evaluation as a targeted radiotherapeutic for the treatment of brain tumors

  15. The fatty acid biosynthesis enzyme FabI plays a key role in the development of liver-stage malarial parasites.

    Science.gov (United States)

    Yu, Min; Kumar, T R Santha; Nkrumah, Louis J; Coppi, Alida; Retzlaff, Silke; Li, Celeste D; Kelly, Brendan J; Moura, Pedro A; Lakshmanan, Viswanathan; Freundlich, Joel S; Valderramos, Juan-Carlos; Vilcheze, Catherine; Siedner, Mark; Tsai, Jennifer H-C; Falkard, Brie; Sidhu, Amar Bir Singh; Purcell, Lisa A; Gratraud, Paul; Kremer, Laurent; Waters, Andrew P; Schiehser, Guy; Jacobus, David P; Janse, Chris J; Ager, Arba; Jacobs, William R; Sacchettini, James C; Heussler, Volker; Sinnis, Photini; Fidock, David A

    2008-12-11

    The fatty acid synthesis type II pathway has received considerable interest as a candidate therapeutic target in Plasmodium falciparum asexual blood-stage infections. This apicoplast-resident pathway, distinct from the mammalian type I process, includes FabI. Here, we report synthetic chemistry and transfection studies concluding that Plasmodium FabI is not the target of the antimalarial activity of triclosan, an inhibitor of bacterial FabI. Disruption of fabI in P. falciparum or the rodent parasite P. berghei does not impede blood-stage growth. In contrast, mosquito-derived, FabI-deficient P. berghei sporozoites are markedly less infective for mice and typically fail to complete liver-stage development in vitro. This defect is characterized by an inability to form intrahepatic merosomes that normally initiate blood-stage infections. These data illuminate key differences between liver- and blood-stage parasites in their requirements for host versus de novo synthesized fatty acids, and create new prospects for stage-specific antimalarial interventions.

  16. Imaging of HER2/neu-positive BT-474 human breast cancer xenografts in athymic mice using {sup 111}In-trastuzumab (Herceptin) Fab fragments

    Energy Technology Data Exchange (ETDEWEB)

    Tang Ying [Division of Nuclear Medicine, University Health Network, Toronto, Ontario, M5G 2C4 (Canada); Department of Pharmaceutical Sciences, University of Toronto, Toronto, Ontario, M5S 2S2 (Canada); Wang, Judy [Division of Nuclear Medicine, University Health Network, Toronto, Ontario, M5G 2C4 (Canada); Scollard, Deborah A. [Division of Nuclear Medicine, University Health Network, Toronto, Ontario, M5G 2C4 (Canada); Mondal, Hridya [Division of Nuclear Medicine, University Health Network, Toronto, Ontario, M5G 2C4 (Canada); Holloway, Claire [Sunnybrook and Women' s College Health Sciences Center, Toronto, Ontario, M4N 3M5 (Canada); Kahn, Harriette J. [Sunnybrook and Women' s College Health Sciences Center, Toronto, Ontario, M4N 3M5 (Canada); Reilly, Raymond M. [Division of Nuclear Medicine, University Health Network, Toronto, Ontario, M5G 2C4 (Canada) and Department of Pharmaceutical Sciences, University of Toronto, Toronto, Ontario, M5S 2S2 (Canada) and Department of Medical Imaging, University of Toronto, Toronto, Ontario, M5S 3E2 (Canada)]. E-mail: raymond.reilly@utoronto.ca

    2005-01-01

    Trastuzumab (Herceptin) Fab were prepared by digestion of intact IgG with immobilized papain, derivatized with diethylenetriaminepentaacetic acid (DTPA) and radiolabeled with {sup 111}In. The dissociation constant (K{sub d}) for binding of Fab to HER2/neu-positive SK-BR-3 human breast cancer cells was two- to threefold higher than for intact IgG (14-36 vs. 8-14 nM). The binding affinity was not significantly decreased after DTPA derivatization (K{sub d}=47 nM). {sup 111}In-trastuzumab Fab localized specifically in HER2/neu-positive BT-474 human breast cancer xenografts in athymic mice with tumor uptake of 7.8{+-}0.7% injected dose (ID)/g and tumor/blood ratio of 25.2{+-}1.6 at 72 h postinjection compared with 2.7{+-}0.7% ID/g and 7.0{+-}0.9 for {sup 111}In-HuM195 anti-CD33 Fab (significantly different, P<.001). Small (3-5 mm in diameter) BT-474 tumors were imaged with {sup 111}In-trastuzumab Fab as early as 24 h postinjection.

  17. Acoustic detection of Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae) and Oryctes elegans (Coleoptera: Scarabaeidae) in Phoenix dactylifera (Arecales: Arecacae) trees and offshoots in Saudi Arabian orchards

    Science.gov (United States)

    Rhynchophorus ferrugineus (Olivier) (Coleoptera: Dryophthoridae) larvae are cryptic, internal-tissue feeding pests of palm trees that are difficult to detect until after they have caused severe economic damage; consequently, infestations may remain undetected until they are widespread in an orchard....

  18. Impact of FAB classification on predicting outcome in acute myeloid leukemia, not otherwise specified, patients undergoing allogeneic stem cell transplantation in CR1: An analysis of 1690 patients from the acute leukemia working party of EBMT.

    Science.gov (United States)

    Canaani, Jonathan; Beohou, Eric; Labopin, Myriam; Socié, Gerard; Huynh, Anne; Volin, Liisa; Cornelissen, Jan; Milpied, Noel; Gedde-Dahl, Tobias; Deconinck, Eric; Fegueux, Nathalie; Blaise, Didier; Mohty, Mohamad; Nagler, Arnon

    2017-04-01

    The French, American, and British (FAB) classification system for acute myeloid leukemia (AML) is extensively used and is incorporated into the AML, not otherwise specified (NOS) category in the 2016 WHO edition of myeloid neoplasm classification. While recent data proposes that FAB classification does not provide additional prognostic information for patients for whom NPM1 status is available, it is unknown whether FAB still retains a current prognostic role in predicting outcome of AML patients undergoing allogeneic stem cell transplantation. Using the European Society of Blood and Bone Marrow Transplantation registry we analyzed outcome of 1690 patients transplanted in CR1 to determine if FAB classification provides additional prognostic value. Multivariate analysis revealed that M6/M7 patients had decreased leukemia free survival (hazard ratio (HR) of 1.41, 95% confidence interval (CI), 1.01-1.99; P = .046) in addition to increased nonrelapse mortality (NRM) rates (HR, 1.79; 95% CI, 1.06-3.01; P = .028) compared with other FAB types. In the NPM1 wt AML, NOS cohort, FAB M6/M7 was also associated with increased NRM (HR, 2.17; 95% CI, 1.14-4.16; P = .019). Finally, in FLT3-ITD + patients, multivariate analyses revealed that specific FAB types were tightly associated with adverse outcome. In conclusion, FAB classification may predict outcome following transplantation in AML, NOS patients. © 2017 Wiley Periodicals, Inc.

  19. A novel antibody-drug conjugate anti-CD19(Fab)-LDM in the treatment of B-cell non-Hodgkin lymphoma xenografts with enhanced anticancer activity.

    Science.gov (United States)

    Jiang, Linlin; Yang, Ming; Zhang, Xiaoyun; Bao, Shiqi; Ma, Li; Fan, Dongmei; Zhou, Yuan; Xiong, Dongsheng; Zhen, Yongsu

    2016-01-01

    Rituximab is widely used in clinical setting for the treatment of B malignant lymphoma and has achieved remarkable success. However, in most patients, the disease ultimately relapses and become resistant to rituximab. To overcome the limitation, there is still a need to find novel strategy for improving therapeutic efficacy. To construct genetically engineered antibody anti-CD19(Fab)-LDM, and verify the anticancer activity targeted toward B-lymphoma. The anticancer activity of anti-CD19(Fab)-LDM in vitro and in vivo was examined. In vitro, the binding activity and internalization of anti-CD19(Fab)-LDP were measured. Using comet assay and apoptosis, the cytotoxicity of energized fusion proteins was observed. From in vivo experiments, targeting of therapeutic effect and anticancer efficacy bythe fusion protein was verified. Data showed that anti-CD19(Fab)-LDM does not only binding the cell surface but is also internalized into the cell. The energized fusion proteins anti-CD19(Fab)-LDM can induce DNA damage. Furthermore, significant in vivo therapeutic efficacy was observed. The present study demonstrated that the genetically engineered antibody anti-CD19(Fab)-LDM exhibited enhanced cytotoxicity compared to LDM alone. One of the most powerful advantages of anti-CD19(Fab)-LDM, however, is that it can be internalized within the cells and carry out cytotoxic effects. Therefore, anti-CD19(Fab)-LDM may be as a useful targeted therapy for B-cell lymphoma.

  20. Brachylophora, a new brachypterous genus of Rhopalophorini (Coleoptera, Cerambycidae

    Directory of Open Access Journals (Sweden)

    Robin O. S. Clarke

    2011-06-01

    Full Text Available Brachylophora, a new brachypterous genus of Rhopalophorini (Coleoptera, Cerambycidae. Brachylophora auricollis (Bruch, 1918 comb. nov. = Pasiphyle auricollis Bruch, 1918, originally described from Argentina (Salta, is redescribed and illustrated. Although with reduced elytra, the genus is transferred from Rhinotragini to Rhopalophorini based on the following characters: eyes well separated in both sexes, frons between eyes depressed and lacking frontal suture; pro-, meso-, and metasternum planar; mesothorax parallel-sided, not at all declivous before mesosternal process; metasternum large, together with mesosternum twice length of prosternum, metepisternum very wide, entire suture separating it from metasternum clearly visible when viewed from below; female ovipositor shortened with short cylindrical styles; and, more generally, structural features of hind legs, and surface ornamentation. Habitus similar to Coremia group. Bolivian specimens were netted as they visited flowers of Croton sp. (Euphorbiaceae.

  1. Diversidad de la familia Carabidae (Coleoptera en Chile Diversity of the family Carabidae (Coleoptera in Chile

    Directory of Open Access Journals (Sweden)

    SERGIO ROIG-JUÑENT

    2001-09-01

    Full Text Available Carabidae constituye dentro de los coléopteros chilenos, la cuarta familia en importancia por su cantidad de especies. El presente trabajo incluye una breve compilación acerca de la historia de la familia y de las primeras expediciones realizadas en Chile. También se realizan comparaciones de la diversidad de carábidos chilenos con respecto a otros países y el Neotrópico. Para Chile, se conocen 21 tribus, con 95 géneros y 365 especies, que representan el 38,8, 28,8 y 7,9 % de la fauna del Neotrópico, respectivamente. Chile posee un bajo número de tribus comparado con otros países, sin embargo, constituye un área importante por la presencia de seis tribus relictuales, principalmente pangeicas o gondwánicas. Chile posee 18 géneros endémicos (18,5 % de su fauna de Carabidae, 28 cuya distribución está restringida a Chile y Argentina y seis restringidos a Chile, Argentina y Uruguay. La cantidad de especies presentes en Chile es inferior a la que poseen otros países de América del Sur, pero la cantidad de especies endémicas es muy alta (204 y representa el 55,8 % de su fauna de carábidos. El alto grado de endemismo que posee Chile con respecto a otros países de América del Sur puede deberse a su condición de aislamiento, siendo las barreras más importantes la región desértica del norte y la cordillera de Los Andes. Este hecho también se vislumbra por la ausencia de importantes tribus neotropicales como Galeritini, Scaritini y Brachinini. También se incluyen en este trabajo claves para la identificación de todas las tribus y géneros presentes en Chile, como así también una breve descripción acerca de la diversidad y ambientes en los que se encuentra cada géneroThe family Carabidae is the fourth largest Coleoptera family in Chile. The present work includes a brief compilation on the taxonomic history of the family and the first expeditions to Chile. In addition, knowledge of carabid diversity in Chile is compared with

  2. Structure of anti-FLAG M2 Fab domain and its use in the stabilization of engineered membrane proteins

    International Nuclear Information System (INIS)

    Roosild, Tarmo P.; Castronovo, Samantha; Choe, Senyon

    2006-01-01

    The X-ray crystallographic analysis of anti-FLAG M2 Fab is reported and the implications of the structure on FLAG epitope binding are described as a first step in the development of a tool for the structural and biophysical study of membrane proteins. The inherent difficulties of stabilizing detergent-solubilized integral membrane proteins for biophysical or structural analysis demand the development of new methodologies to improve success rates. One proven strategy is the use of antibody fragments to increase the ‘soluble’ portion of any membrane protein, but this approach is limited by the difficulties and expense associated with producing monoclonal antibodies to an appropriate exposed epitope on the target protein. Here, the stabilization of a detergent-solubilized K + channel protein, KvPae, by engineering a FLAG-binding epitope into a known loop region of the protein and creating a complex with Fab fragments from commercially available anti-FLAG M2 monoclonal antibodies is reported. Although well diffracting crystals of the complex have not yet been obtained, during the course of crystallization trials the structure of the anti-FLAG M2 Fab domain was solved to 1.86 Å resolution. This structure, which should aid future structure-determination efforts using this approach by facilitating molecular-replacement phasing, reveals that the binding pocket appears to be specific only for the first four amino acids of the traditional FLAG epitope, namely DYKD. Thus, the use of antibody fragments for improving the stability of target proteins can be rapidly applied to the study of membrane-protein structure by placing the short DKYD motif within a predicted peripheral loop of that protein and utilizing commercially available anti-FLAG M2 antibody fragments

  3. Engineering an improved IgG4 molecule with reduced disulfide bond heterogeneity and increased Fab domain thermal stability.

    Science.gov (United States)

    Peters, Shirley J; Smales, C Mark; Henry, Alistair J; Stephens, Paul E; West, Shauna; Humphreys, David P

    2012-07-13

    The integrity of antibody structure, stability, and biophysical characterization are becoming increasingly important as antibodies receive increasing scrutiny from regulatory authorities. We altered the disulfide bond arrangement of an IgG4 molecule by mutation of the Cys at the N terminus of the heavy chain constant domain 1 (C(H)1) (Kabat position 127) to a Ser and introduction of a Cys at a variety of positions (positions 227-230) at the C terminus of C(H)1. An inter-LC-C(H)1 disulfide bond is thus formed, which mimics the disulfide bond arrangement found in an IgG1 molecule. The antibody species present in the supernatant following transient expression in Chinese hamster ovary cells were analyzed by immunoblot to investigate product homogeneity, and purified product was analyzed by a thermofluor assay to determine thermal stability. We show that the light chain can form an inter-LC-C(H)1 disulfide bond with a Cys when present at several positions on the upper hinge (positions 227-230) and that such engineered disulfide bonds can consequently increase the Fab domain thermal stability between 3 and 6.8 °C. The IgG4 disulfide mutants displaying the greatest increase in Fab thermal stability were also the most homogeneous in terms of disulfide bond arrangement and antibody species present. Importantly, mutations did not affect the affinity for antigen of the resultant molecules. In combination with the previously described S241P mutation, we present an IgG4 molecule with increased Fab thermal stability and reduced product heterogeneity that potentially offers advantages for the production of IgG4 molecules.

  4. Manufacture of β-TCP/alginate scaffolds through a Fab@home model for application in bone tissue engineering

    International Nuclear Information System (INIS)

    Diogo, G S; Gaspar, V M; Serra, I R; Fradique, R; Correia, I J

    2014-01-01

    The growing need to treat bone-related diseases in an elderly population compels the development of novel bone substitutes to improve patient quality of life. In this context, the advent of affordable and effective rapid prototyping equipment, such as the Fab@home plotter, has contributed to the development of novel scaffolds for bone tissue engineering. In this study, we report for the first time the use of a Fab@home plotter for the production of 3D scaffolds composed by beta-tricalcium phosphate (β-TCP)/alginate hybrid materials. β-TCP/alginate mixtures were used in a proportion of 50/50% (w/w), 30/70% (w/w) and 20/80% (w/w). The printing parameters were optimized to a nozzle diameter of 20 Gauge for the production of rigid scaffolds with pre-defined architectures. We observed that, despite using similar printing parameters, both the precision and resolution of the scaffolds were significantly affected by the blend's viscosity. In particular, we demonstrate that the higher viscosity of 50/50 scaffolds (150.0 ± 3.91 mPa s) provides a higher precision in the extrusion process. The physicochemical and biological characterization of the samples demonstrated that the 50/50 scaffolds possessed a resistance to compression comparable to that of native trabecular bone. Moreover, this particular formulation also exhibited a Young's modulus that was higher than that of trabecular bone. Scanning electron microscopy and fluorescence microscopy analysis revealed that osteoblasts were able to adhere, proliferate and also penetrate into the scaffold's architecture. Altogether, our findings suggest that the Fab@home printer can be employed in the manufacture of reproducible scaffolds, using a formulation 50/50 alginate-β-TCP that has suitable properties to be applied as bone substitutes in the future. (paper)

  5. Glomerular filtration rate after alpha-radioimmunotherapy with 211At-MX35-F(ab')2: a long-term study of renal function in nude mice

    DEFF Research Database (Denmark)

    Back, T.; Haraldsson, B.; Hultborn, R

    2009-01-01

    of the glomerular filtration rate (GFR). The renal toxicity was evaluated at levels close to the dose limit for the bone marrow and well within the range for therapeutic efficacy on tumors. Astatinated MX35-F(ab')(2) monoclonal antibodies were administered intravenously to nude mice. Both non-tumor-bearing animals...... manifested late. Examination of the kidney sections showed histologic changes that were overall subdued. Following alpha-RIT with (211)At-MX35-F(ab')(2) at levels close to the dose limit of severe myelotoxicity, the effects found on renal function were relatively small, with only minor to moderate reductions...... in GFR. These results suggest that a mean absorbed dose to the kidneys of approximately 10 Gy is acceptable, and that the kidneys would not be the primary dose-limiting organ in systemic alpha-RIT when using (211)At-MX35-F(ab')(2) Udgivelsesdato: 2009/12...

  6. Pre-existing IgG antibodies cross-reacting with the Fab region of infliximab predict efficacy and safety of infliximab therapy in inflammatory bowel disease

    DEFF Research Database (Denmark)

    Steenholdt, Casper; Palarasah, Yaseelan; Bendtzen, Klaus

    2013-01-01

    are common and may cross-react with the murine part of IFX. AIM: To investigate if Abs binding to IFX's Fab region (IFX-Fab) are present in IBD patients before exposure to IFX, and whether they predict efficacy and safety of IFX therapy. METHODS: Observational, retrospective cohort study of patients with CD...... (n = 29) and UC (n = 22). RESULTS: Pre-treatment levels of IFX-Fab reactive IgG Abs were significantly lower in CD patients in remission after 1 year of maintenance IFX (median 91 mU/L, n = 8) than in the rest of the patients (639 mU/L, n = 21; P ...

  7. Purification, crystallization and preliminary X-ray diffraction analysis of enoyl-acyl carrier protein reductase (FabK) from Streptococcus mutans strain UA159

    International Nuclear Information System (INIS)

    Kim, Tae-O; Im, Dong-Won; Jung, Ha Yun; Kwon, Seong Jung; Heo, Yong-Seok

    2012-01-01

    Enoyl-acyl carrier protein reductase (FabK) from S. mutans strain UA159 was cloned, overexpressed, purified and crystallized. X-ray diffraction data were collected to a resolution of 2.40 Å. A triclosan-resistant flavoprotein termed FabK is the sole enoyl-acyl carrier protein reductase in Streptococcus pneumoniae and Streptococcus mutans. In this study, FabK from S. mutans strain UA159 was overexpressed in Escherichia coli, purified and crystallized. Diffraction data were collected to 2.40 Å resolution using a synchrotron-radiation source. The crystal belonged to space group P6 2 , with unit-cell parameters a = b = 105.79, c = 44.15 Å. The asymmetric unit contained one molecule, with a corresponding V M of 2.05 Å 3 Da −1 and a solvent content of 39.9%

  8. Walking stability of Rhyzopertha dominica (Fabricius, 1792 (Coleoptera: Bostrichidae

    Directory of Open Access Journals (Sweden)

    E. M. Pires

    Full Text Available Abstract Results obtained in studies can contribute to the advancement of science and innovative methods and techniques for developing practical activities. Reporting conditions that may restrict the implementation of research is critical to ensure the optimal development of further technical studies. The objective of this study was to assess the walking stability of R. dominica on a flat and smooth surface. The study was based on the determination of mortality, morphology and walking stability of the insect outside the grain mass, on a flat and smooth surface. Mortality of adults of this Coleoptera in conditions with and without food was similar, which explains the difficulty that this insect had for accessing the food source on the flat and smooth surface. The measurements of body length (BOL, width (BOW and height (BOH of R. dominica were compared with those of Tribolium castaneum (Coleoptera: Tenebrionidae, which showed good ability to walk in these conditions. This study indicated that the former presents lower BOL and BOW, and greater BOH than the second, and all these variables showed differences when analyzed simultaneously by means of the construction of multivariate morphometric indices (Width × Height, Length × Height and Height × Length × Width. These morphometric variables, together with the definition of the geometry most similar to the body shape, resulted in determination of the center of gravity (CG and static rollover threshold (SRTgeom for both species. Rhyzopertha dominica and T. castaneum presented CGs considered high and low, respectively, and together with the values obtained for SRTgeom, may justify that R. dominica can be considered a less stable species during movement, and presents greater risk of rollover on flat and smooth surfaces.

  9. Coleoptera associated with macrophytes of the genus Salvinia in four oxbow lakes in two river basins in southeast Brazil

    Directory of Open Access Journals (Sweden)

    M. C. Paula-Bueno

    Full Text Available Abstract Macrophytes in oxbow lakes represent an important substrate for the Coleoptera. Two oxbow lakes the Rio Paranapanema were studied and the other two Rio Mogi-Guaçu, in the State de São Paulo, Brasil. In this study, there is greater similarity between the communities of Coleoptera of lakes greater connectivity with the main river channel or the difference in the species of Salvinia collected in the lakes studied interferes Coleoptera fauna that uses as substrate. A total of 9,222 specimens of Coleoptera were collected and identified in 10 families and 40 genera. The analysis MDS for abundance of Coleoptera showed the grouping of the oxbow lakes the Paranapanema River and a distancing the oxbow lakes the Mogi-Guaçu. The PERMANOVA test did not reveal any difference in the fauna between the wet and dry periods. It was concluded that the connectivity between river and lake is not decisive for the richness and abundance of aquatic fauna of Coleoptera. Therefore, the richness and abundance of aquatic Coleoptera associated vary with the species of Salvinia used as substrate.

  10. Coleoptera associated with macrophytes of the genus Salvinia in four oxbow lakes in two river basins in southeast Brazil.

    Science.gov (United States)

    Paula-Bueno, M C; Fonseca-Gessner, A A

    2015-11-01

    Macrophytes in oxbow lakes represent an important substrate for the Coleoptera. Two oxbow lakes the Rio Paranapanema were studied and the other two Rio Mogi-Guaçu, in the State de São Paulo, Brasil. In this study, there is greater similarity between the communities of Coleoptera of lakes greater connectivity with the main river channel or the difference in the species of Salvinia collected in the lakes studied interferes Coleoptera fauna that uses as substrate. A total of 9,222 specimens of Coleoptera were collected and identified in 10 families and 40 genera. The analysis MDS for abundance of Coleoptera showed the grouping of the oxbow lakes the Paranapanema River and a distancing the oxbow lakes the Mogi-Guaçu. The PERMANOVA test did not reveal any difference in the fauna between the wet and dry periods. It was concluded that the connectivity between river and lake is not decisive for the richness and abundance of aquatic fauna of Coleoptera. Therefore, the richness and abundance of aquatic Coleoptera associated vary with the species of Salvinia used as substrate.

  11. Clinical trial of an F(ab')(2) polyvalent equine antivenom for African snake bites in Benin

    OpenAIRE

    Chippaux, Jean-Philippe; Massougbodji, A.; Stock, R. P.; Alagon, A.

    2007-01-01

    We report the results of a trial designed to measure the safety and efficacy of African Antivipmyn((R)), a new freeze-dried polyvalent equine F(ab')(2)-based antivenom. We tested 289 envenomations. After treatment, 19% of treated patients had undesirable events, all benign. A possible adverse effect was attributed to this antivenom in 11% of the patients. Bleeding was observed in 48% of the patients; it stopped within 2 hours after treatment with antivenom in 60% of the patients. Blood incoag...

  12. Diversity of forensic rove beetles (Coleoptera, Staphylinidae) associated with decaying pig carcass in a forest biotope.

    Science.gov (United States)

    Dekeirsschieter, Jessica; Frederick, Christine; Verheggen, Francois J; Drugmand, Didier; Haubruge, Eric

    2013-07-01

    Most forensic studies are focused on Diptera pattern colonization while neglecting Coleoptera succession. So far, little information is available on the postmortem colonization by beetles and the decomposition process they initiate under temperate biogeoclimatic countries. These beetles have, however, been referred to as being part of the entomofaunal colonization of a dead body. Forensic entomologists need increased databases detailing the distribution, ecology, and phenology of necrophagous insects, including staphylinids (Coleoptera, Staphylinidae). While pig carcasses are commonly used in forensic entomology studies to surrogate human decomposition and to investigate the entomofaunal succession, very few works have been conducted in Europe on large carcasses. Our work reports the monitoring of the presence of adult rove beetles (Coleoptera, Staphylinidae) on decaying pig carcasses in a forest biotope during four seasons (spring, summer, fall, and winter). A total of 23 genera comprising 60 species of rove beetles were collected from pig carcasses. © 2013 American Academy of Forensic Sciences.

  13. [Infestation of the human digestive system with beetle larvae (Coleoptera: Cantharidae): a case report].

    Science.gov (United States)

    Yılmaz, Hasan; Taş Cengiz, Zeynep; Dülger, Ahmet Cumhur; Ekici, Pınar

    2014-12-01

    This study was conducted to report the digestive system infestation caused by the larvae of Coleoptera in a female pediatric patient. She was admitted to our hospital with the complaints of emergence of insect larvae from her vomit and feces, abdominal pain, inguinal pain, lack of appetite, hair loss, excessive cleaning behavior, extreme irritability, and distractibility. The larvae observed typically had the morphology of the larvae of insects related to the Cantharidae family in the Coleoptera order. For treatment, a single dose of albendazole (400 mg) was used. Consequently, in the present case, it was seen that the larvae of Coleoptera incidentally taken orally could continue to live for a period in the digestive tract of people, without losing vitality, and the larvae caused a variety of symptoms due to both their toxic agents and the possible irritation they caused.

  14. Pharmacological efficacy of anti-IL-1β scFv, Fab and full-length antibodies in treatment of rheumatoid arthritis.

    Science.gov (United States)

    Qi, Jianying; Ye, Xianlong; Ren, Guiping; Kan, Fangming; Zhang, Yu; Guo, Mo; Zhang, Zhiyi; Li, Deshan

    2014-02-01

    Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease that mainly causes the synovial joint inflammation and cartilage destruction. Interleukin-1β (IL-1β) is an important proinflammatory cytokine involved in the pathogenesis of RA. In this study, we constructed and expressed anti-IL-1β-full-length antibody in CHO-K1-SV, anti-IL-1β-Fab and anti-IL-1β-scFv in Rosetta. We compared the therapeutic efficacy of three anti-IL-1β antibodies for CIA mice. Mice with CIA were subcutaneously injected with humanized anti-IL-1β-scFv, anti-IL-1β-Fab or anti-IL-1β-full-length antibody. The effects of treatment were determined by arthritis severity score, autoreactive humoral, cellular immune responses, histological lesion and cytokines production. Compared with anti-IL-1β-scFv treatments, anti-IL-1β-Fab and anti-IL-1β-full-length antibody therapy resulted in more significant effect in alleviating the severity of arthritis by preventing bone damage and cartilage destruction, reducing humoral and cellular immune responses, and down-regulating the expression of IL-1β, IL-6, IL-2, IFN-γ, TNF-α and MMP-3 in inflammatory tissue. The therapeutic effects of anti-IL-1β-Fab and anti-IL-1β-full-length antibodies on CIA mice had no significant difference. However, production of anti-IL-1β-full-length antibody in eukaryotic system is, in general, time-consuming and more expensive than that of anti-IL-1β-Fab in prokaryotic systems. In conclusion, as a small molecule antibody, anti-IL-1β-Fab is an ideal candidate for RA therapy. Copyright © 2013 Elsevier Ltd. All rights reserved.

  15. Sortase-catalyzed in vitro functionalization of a HER2-specific recombinant Fab for tumor targeting of the plant cytotoxin gelonin

    Science.gov (United States)

    Kornberger, Petra; Skerra, Arne

    2014-01-01

    We report on the preparation of a new type of immunotoxin via in vitro ligation of the αHer2 antigen binding fragment (Fab) of the clinically-validated antibody trastuzumab to the plant toxin gelonin, employing catalysis by the bacterial enzyme sortase A (SrtA). The αHer2 Fab was fused with the extended SrtA recognition motif LPET↓GLEH6 at the C-terminus of its heavy chain, thereby preventing interference with antigen binding, while the toxin was equipped with a Gly2 sequence at its N-terminus, distant to the catalytically active site in the C-terminal region. Site-specific in vitro transpeptidation led to a novel antibody-toxin conjugate wherein gelonin had effectively replaced the Fc region of a conventional (monomerized) immunoglobulin. After optimization of reaction conditions and incubation time, the resulting Fab-Gelonin ligation product was purified to homogeneity in a two-step procedure by means of Strep-Tactin affinity chromatography—utilizing the Strep-tag II appended to gelonin—and size exclusion chromatography. Binding activity of the immunotoxin for the Her2 ectodomain was indistinguishable from the unligated Fab as measured by real-time surface plasmon resonance spectroscopy. Specific cytotoxic potency of Fab-Gelonin was demonstrated against two Her2-positive cell lines, resulting in EC50 values of ~1 nM or lower, indicating a 1000-fold enhanced cell-killing activity compared with gelonin itself. Thus, our strategy provides a convenient route to the modular construction of functional immunotoxins from Fabs of established tumor-specific antibodies with gelonin or related proteotoxins, also avoiding the elevated biosafety levels that would be mandatory for the direct biotechnological preparation of corresponding fusion proteins. PMID:24492291

  16. Safety, pharmacokinetic and dosimetry evaluation of the proposed thrombus imaging agent 99mTc-DI-DD-3B6/22-80B3 Fab'

    International Nuclear Information System (INIS)

    Macfarlane, David J.; Smart, Richard C.; Tsui, Wendy W.; Gerometta, Michael; Eisenberg, Paul R.; Scott, Andrew M.

    2006-01-01

    99m Tc-DI-DD-3B6/22-80B3 (Thromboview, hereafter abbreviated to 99m Tc-DI-80B3 Fab') is a humanised, radiolabelled monoclonal antibody Fab' fragment with high affinity and specificity for the D-dimer domain of cross-linked fibrin. The purpose of this study was to evaluate the safety, pharmacokinetics and dosimetry of four increasing doses of 99m Tc-DI-80B3 Fab' in healthy volunteers. Thirty-two healthy volunteers (18-70 years; 16 male, 16 female) received a single intravenous injection of 0.5, 1.0, 2.0 or 4.0 mg of 99m Tc-DI-80B3 Fab'. Safety outcomes (vital signs, electrocardiography, haematology, biochemistry, adverse events and development of human anti-human antibodies) were assessed up to 30 days post injection. Blood and urine samples were collected up to 48 h post injection. Gamma camera images were acquired at 0.5, 1, 2, 4, 6 and 24 h post injection. Dosimetry was performed using standard MIRD methodology. No adverse events considered to be drug related were observed. Human anti-human antibody was not detectable in any subject during the follow-up period. 99m Tc-DI-80B3 Fab' had a rapid initial plasma clearance (t 1/2 α=1 h). The pharmacokinetic profile of the Fab' fragment was generally linear across the four dose cohorts. By 24 h, 30-35% of the administered radioactivity appeared in the urine. There was marked renal accumulation with time, but no specific uptake was identified within other normal tissues. The effective dose was 9 mSv/750 MBq. (orig.)

  17. Modified cytokeratins expressed on the surface of carcinoma cells undergo endocytosis upon binding of human monoclonal antibody and its recombinant Fab fragment

    DEFF Research Database (Denmark)

    Ditzel, H J; Garrigues, U; Andersen, C B

    1997-01-01

    display selection and the human Fab fragment was expressed in bacteria. Analysis by confocal laser scanning microscopy demonstrated that COU-1 bound in a uniform punctate pattern to the surface of viable carcinoma cells stained at 4 degrees C, and binding increased significantly when cells were cultured...... was significantly reduced. Similar results were obtained using intact IgM COU-1 and the recombinant Fab fragment. Immunohistological studies indicated that COU-1, in contrast to murine monoclonal antibodies against normal cytokeratin 8 and 18, could differentiate between malignant and normal colon epithelia...

  18. Integration of Gamma Irradiation and Some Botanical oils To Protect Cowpea And Chickpea Seeds From Infestation With The Bruchid Beetle Callosobruchus Maculatus

    International Nuclear Information System (INIS)

    Hassan, R.S.; Mikhaiel, A.A.; Sileem, Th.M.

    2013-01-01

    The lethal effect of gamma radiation doses of 0.75 or 1.0 kGy on the adults Callosobruchus maculates reared on cowpeas and chickpeas were slow during the first and third days post-treatments. By increasing the dose to 1.5 kGy, the values of the percent mortality of adults in both seeds 24 h posttreatment were 53 and 40%, respectively. On the other hand, the dose 2 kGy caused sooner mortality for adults post-treatment for cowpeas. Different concentrations from eight plant oils; lemon grass, pinus sylvestris, parsley, fennel, geranium, peppermint, petitgrain and sweet basil, were used for protection of cowpea and chickpea seeds from infestation by Callosobruchus maculates. The results showed that sweet basil and geranium caused 89 and 79 % larval mortality, respectively, in case of cowpeas at concentration 0.5 % with exposure period of 48 hour while 71.0 and 63.33% adult mortality was occurred at the same concentration of both oils in chickpeas. The latent effects of tested plant oils on adult stage when beetles of C. maculatus were fed on seeds treated with the lowest two concentrations (0.0312, 0.0625%) of tested oils, the number of eggs laid per female was decreased in female exposed to all tested oils especially in case of cowpea treated with sweet basil and lemongrass. Most of the tested oils caused high reduction in larval penetration in both types of seeds. The adult weight was non significantly reduced at all treatments. The use of different plant oils leads to reduction in the progeny comparing to the control and sweet basil or geranium was found to be highly effective in decreasing the percentage of emergence (30 and 40% , respectively). No harmful effect was observed on germination of plant oils treated cowpea and chickpea seeds with concentration 2%.

  19. Protecting embryos from stress: Corticosterone effects and the corticosterone response to capture and confinement during pregnancy in a live-bearing lizard (Hoplodactylus maculatus)

    Science.gov (United States)

    Cree, A.; Tyrrell, C.L.; Preest, M.R.; Thorburn, D.; Guillette, L.J.

    2003-01-01

    Hormones in the embryonic environment, including those of the hypothalamo-pituitary-adrenal (HPA) axis, have profound effects on development in eutherian mammals. However, little is known about their effects in reptiles that have independently evolved viviparity. We investigated whether exogenous corticosterone affected embryonic development in the viviparous gecko Hoplodactylus maculatus, and whether pregnant geckos have a corticosterone response to capture and confinement that is suppressed relative to that in non-pregnant (vitellogenic) females and males. Corticosterone implants (5 mg, slow-release) administered to females in mid-pregnancy caused a large elevation of corticosterone in maternal plasma (P<0.001), probable reductions in embryonic growth and development (P=0.069-0.073), developmental abnormalities and eventual abortions. Cool temperature produced similar reductions in embryonic growth and development (P???0.036 cf. warm controls), but pregnancies were eventually successful. Despite the potentially harmful effects of elevated plasma corticosterone, pregnant females did not suppress their corticosterone response to capture and confinement relative to vitellogenic females, and both groups of females had higher responses than males. Future research should address whether lower maternal doses of corticosterone produce non-lethal effects on development that could contribute to phenotypic plasticity. Corticosterone implants also led to increased basking in pregnant females (P<0.001), and basal corticosterone in wild geckos (independent of reproductive condition) was positively correlated with body temperature (P<0.001). Interactions between temperature and corticosterone may have broad significance to other terrestrial ectotherms, and body temperature should be considered as a variable influencing plasma corticosterone concentrations in all future studies on reptiles. ?? 2003 Elsevier Inc. All rights reserved.

  20. Effects of Thermal Stress on the mRNA Expression of SOD, HSP90, and HSP70 in the Spotted Sea Bass ( Lateolabrax maculatus)

    Science.gov (United States)

    Shin, Moon-Kyeong; Park, Ho-Ra; Yeo, Won-Jun; Han, Kyung-Nam

    2018-03-01

    The aim of this study was to elucidate the molecular mechanisms underlying the thermal stress response in the spotted sea bass ( Lateolabrax maculatus). Spotted sea basses were exposed to 4 different water temperatures (20, 22, 24, and 28°C) in increasing increments of 2°C/h from 18°C (control) for different time periods (0, 6, 12, 24, 48, 72, and 96 h). Subsequently, 3 tissues (liver, muscle, and gill) were isolated, and the levels of SOD, HSP90, and HSP70 mRNA were assessed. SOD mRNA expression was maintained at baseline levels of control fish at all water temperatures in the liver, while muscle and gill tissue showed an increase followed by a decrease over each certain time with higher water temperature. HSP90 mRNA expression increased in the liver at ≤ 24°C over time, but maintained baseline expression at 28°C. In muscle, HSP90 mRNA expression gradually increased at all water temperatures, but increased and then decreased at ≥ 24°C in gill tissue. HSP70 mRNA expression exhibited an increase and then a decrease in liver tissue at 28°C, but mainly showed similar expression patterns to HSP90 in all tissues. These results suggest the activity of a defense mechanism using SOD, HSP90, and HSP70 in the spotted sea bass upon rapid increases in water temperature, where the expression of these genes indicated differences between tissues in the extent of the defense mechanisms. Also, these results indicate that high water temperature and long-term thermal stress exposure can inhibit physiological defense mechanisms.

  1. Biomarkers and bioindicators of the environmental condition using a fish species (Pimelodus maculatus Lacepède, 1803 in a tropical reservoir in Southeastern Brazil

    Directory of Open Access Journals (Sweden)

    F. G. Araújo

    2017-08-01

    Full Text Available Abstract The Funil Reservoir receives a large amount of xenobiotics from the Paraíba do Sul River (PSR from large number of industries and municipalities in the watershed. This study aimed to assess environmental quality along the longitudinal profile of the Paraíba do Sul River–Funil Reservoir system, by using biomarkers and bioindicators in a selected fish species. The raised hypothesis is that Funil Reservoir acts as a filter for the xenobiotics of the PSR waters, improving river water quality downstream the dam. Two biomarkers, the ethoxyresorufin–O–deethylase activity (EROD, measured as fluorimetricly in S9 hepatic fraction, and the micronuclei frequency (MN, observed in erythrocytes of the cytoplasm, and three bioindicators, the hepatosomatic index (HSI, gonadosomatic index (GSI and condition factor (CF were used in Pimelodus maculatus, a fish species widely distributed in the system. Four zones were searched through a longitudinal gradient: 1, river upstream from the reservoir; 2, upper reservoir; 3, lower reservoir; 4, river downstream of the reservoir. EROD activity and HSI and GSI had significant differences among the zones (P<0.05. The upper reservoir had the lowest EROD activity and HSI, whereas the river downstream of the reservoir had the highest EROD and lowest GSI. The river upstream from the reservoir showed the highest HSI and GSI. It is suggested that the lowest environmental condition occur at the river downstream of the reservoir, where it seems to occur more influence of xenobiotics, which could be associated with hydroelectric plant operation. The hypothesis that Funil reservoir acts as a filter decanting pollution from the Paraíba do Sul River waters was rejected. These results are novel information on this subject for a native fish species and could be useful for future comparisons with other environments.

  2. Bio-edafology of the Coleoptera order, in three Colombia Natural Regions

    International Nuclear Information System (INIS)

    Camero R, Edgar; Chamorro B, Clara

    1997-01-01

    The characterization of Coleoptera families is showed to three different Colombian Natural Regions. This is given in relation with biological and environmental factors, different vegetation covers and soil uses. In addition to it, susceptible Coleoptera taxa are determined when natural conditions are disturbed. Methodically, a literature subject research was made, and Barber and Berlesse traps were used to organism extractions from superficial and under superficial soil. Horizons diversity, riches and constancy index were determined to each family. Results show different diversity, riches and constancy values to each family, as much in each natural region, such as to each soil use

  3. Controle do caruncho Callosobruchus maculatus (Fabr., 1775 (Coleóptera: Bruchidae utilizando extratos de Piper nigrum L. (Piperaceae pelo método de vapor Control of cowpea weevil Callosobruchus maculatus (Fabr., 1775(Coleóptera: Bruchidae using extracts of Piper nigrum L. (Piperaceae by the steam method

    Directory of Open Access Journals (Sweden)

    Silvana Alves de Almeida

    2006-08-01

    Full Text Available Considerando os prejuízos causados pelo inseto-praga Callosobruchus maculatus (Fabr., 1775 ao feijão Vigna unguiculata Walp. armazenado, com o presente trabalho objetivou-se avaliar a eficácia de extratos de Piper nigrum L. na mortalidade de adultos daquela espécie. O delineamento estatístico utilizado foi inteiramente ao acaso com quatro repetições e arranjo fatorial 3 x 5, constituído por três concentrações do extrato e cinco períodos de exposições dos extratos (5, 10, 15, 20 e 25 minutos. Utilizaram-se frutos secos triturados de P. nigrum para extração em percolador com solvente álcool etílico (70, 50 e 30%. Os extratos foram aplicados na forma de vapor, por meio de um compressor adaptado, para dentro de recipiente contendo 100 insetos. Os resultados permitiram concluir que a mortalidade dos insetos aumenta com o aumento do período de exposição aos extratos, e que todas as concentrações se mostraram eficientes, embora, em termos de valores absolutos, o extrato com 70% de álcool etílico foi o mais eficaz.Considering the damage caused by the insect pest Callosobruchus maculatus (Fabr., 1775 to stored Vigna unguiculata Walp beans, this work aimed to evaluate the effectiveness of extracts of Piper nigrum L. in the control of adults of the referred species. The statistical outline used was made at random with four repetitions and factorial arrangement 3 x 5, formed by three concentrations of the extract and five periods of exposure to the extracts (5, 10, 15, 20 and 25 minutes. P. nigrum grounded dry fruits were used for extraction , which was done with ethyl alcohol (70, 50 and 30%, using a percolator. The extracts were applied as steam, using an adapted compressor into a container having 100 insects. By the results it was possible to conclude that the mortality of the insects becomes higher as the period of exposure to the extracts is increased, and that all the concentrations were shown to be effective, although, in

  4. UTILISATION DE POUDRES ET D'EXTRAITS TOTAUX ISSUS DE ...

    African Journals Online (AJOL)

    AISA

    EXTRAITS TOTAUX. ISSUS DE PLANTES LOCALES DES GENRES Ocimum sp. ET Mentha sp. COMME BIOPESTICIDES DANS LA LUTTE. CONTRE Callosobruchus maculatus FAB. F. JOHNSON, B. SERI-KOUASSI, L. R. N. ABOUA, K. FOUA-BI.

  5. The role of indium-111 antimyosin (Fab) imaging as a noninvasive surveillance method of human heart transplant rejection

    International Nuclear Information System (INIS)

    De Nardo, D.; Scibilia, G.; Macchiarelli, A.G.

    1989-01-01

    The identification of rejection after heart transplantation in patients receiving cyclosporine immunosuppressive therapy requires the endomyocardial biopsy, an invasive method associated with a finite morbidity. To evaluate the role of indium-111 antimyosin (Fab) scintigraphy as a noninvasive surveillance method of heart transplant rejection, the Fab fragment of murine monoclonal antimyosin antibodies labeled with indium-111 was administered intravenously in 30 scintigraphic studies to 10 consecutive heart transplant recipients. Endomyocardial biopsy specimens were obtained 72 hours after each scintigraphic study. Nineteen scintigraphic studies had negative findings; no false negative finding was obtained. Eleven antimyosin scintigraphic studies had positive findings, and in these studies endomyocardial biopsy revealed mild rejection in two cases, moderate acute rejection with myocyte necrosis in two cases, myocyte necrosis as a consequence of ischemic injury in six cases, and possibly cytotoxic damage in one case. Antimyosin scintigraphy may represent a reliable screening method for the surveillance of heart transplant patients. In the presence of a negative finding from antimyosin scintigraphy, it may be possible to avoid endomyocardial biopsy. Conversely, in patients who have a positive finding from antimyosin scintigraphy, the endomyocardial biopsy is mandatory to establish the definitive diagnosis by histologic examination of the myocardium

  6. Serum and urine analysis of the aminoterminal procollagen peptide type III by radioimmunoassay with antibody Fab fragments.

    Science.gov (United States)

    Rohde, H; Langer, I; Krieg, T; Timpl, R

    1983-09-01

    A radioimmunoassay based on antibody Fab fragments was developed for the aminoterminal peptide Col 1-3 of bovine type III procollagen. This assay does not distinguish the intact aminopropeptide Col 1-3 from its globular fragment Col 1. Parallel inhibition profiles were observed with human serum and urine allowing the simultaneous quantitative determination of intact and fragmented antigens in these samples. Most of the material has a size similar to that of fragment Col 1 indicating that the aminopropeptide is degraded under physiologic conditions. The concentration of aminopeptide in normal sera was in the range 15-63 ng/ml. Daily excretion was found to be in the range 30-110 micrograms. More than 50% of patients with alcoholic hepatitis and liver cirrhosis showed elevated serum levels of aminopropeptide by the Fab assay. Elevated concentrations were detected more frequently with an antibody radioimmunoassay which measures mainly the intact form of the aminopropeptide. It is suggested that analysis of patients material by both assays could improve their diagnostic application.

  7. F(ab'2 antibody fragments against Trypanosoma cruzi calreticulin inhibit its interaction with the first component of human complement

    Directory of Open Access Journals (Sweden)

    LORENA AGUILAR

    2005-01-01

    Full Text Available Trypanosoma cruzi calreticulin (TcCRT, described in our laboratory, retains several important functional features from its vertebrate homologues. We have shown that recombinant TcCRT inhibits the human complement system when it binds to the collagenous portion of C1q. The generation of classical pathway convertases and membrane attack complexes is thus strongly inhibited. In most T. cruzi-infected individuals, TcCRT is immunogenic and mediates the generation of specific antibodies. By reverting the C1q / TcCRT interaction, a parasite immune evasion strategy, these antibodies contribute to the host / parasite equilibrium. In an in vitro correlate of this situation, we show that the C1q / TcCRT interaction is inhibited by F(ab'2 polyclonal anti-TcCRT IgG fragments. It is therefore feasible that in infected humans anti-TcCRT antibodies participate in reverting an important parasite strategy aimed at inhibiting the classical complement pathway. Thus, membrane-bound TcCRT interacts with the collagenous portion C1q, and this C1q is recognized by the CD91-bound host cell CRT, thus facilitating parasite internalization. Based on our in vitro results, it could be proposed that the in vivo interaction between TcCRT and vertebrate C1q could be inhibited by F(ab'2 fragments anti-rTcCRT or against its S functional domain, thus interfering with the internalization process

  8. Influence of cage farming on feeding and reproductive aspects of Pimelodus maculatus Lacépède, 1803 (Siluriformes: Pimelodidae in the Chavantes reservoir, Brazil - doi: 10.4025/actascibiolsci.v36i1.

    Directory of Open Access Journals (Sweden)

    Heleno Brandão

    2013-09-01

    Full Text Available This study evaluated the diet and reproductive aspects of the population of Pimelodus maculatus around net cage fish farming in order to assess the possible impacts of this activity. Monthly collections were performed from March 2008 to February 2009 on two populations: one close to the net cages (NC and one from an area not influenced by these cages denominated the “reference site” (RS. Results of the Alimentary Index (AI, Gonadosomatic Index (GSI, reproductive potential and histological analysis were obtained for both NC and RS populations. The population from NC used leftover food (ration that escapes from net cages as the main food item (99.3%. For the RS population, the detritus item was the more important food source (51.7%. The Detrended Correspondence Analysis (DCA showed that the use of food resources was different between the two sites. The reproductive period of the species (indicated by the GSI revealed that the population of the NC showed an extended reproductive period compared to RS. The histology of the ovaries indicated that the specimens in the NC were spawning capable. This study indicates that fish farming activities influence the species P. maculatus in the Chavantes reservoir by adding a new resource to the food web.

  9. Responses by Dendroctonus frontalis and Dendroctonus mesoamericanus (Coleoptera: Curculionidae) to Ssemiochemical lures in Chiapas, Mexico: possible roles of pheromones during joint host attacks

    Science.gov (United States)

    Alicia Nino-Dominguez; Brian T. Sullivan; Jose H. Lopez-Urbina; Jorge E. Macias-Samano

    2016-01-01

    In southern Mexico and Central America, the southern pine beetle Dendroctonus frontalis Zimmermann (Coleoptera: Curculionidae: Scolytinae) commonly colonizes host trees simultaneously with Dendroctonus mesoamericanus Armend

  10. Susceptibility Of Five Cowpea ( Vigna unguiculata ) Varieties To ...

    African Journals Online (AJOL)

    Susceptibility Of Five Cowpea ( Vigna unguiculata ) Varieties To Attack By Callosobruchus maculatus (Fab.) ... in Ghana were screened for susceptibility to the cowpea beetle, C. maculatus, in storage for a period of 30 days in the laboratory at an ambient temperature of 27.5 ± 0.9oC and relative humidity of 64.6 ± 2%.

  11. Role of emerald ash borer (Coleoptera: Buprestidae) larval vibrations in host-quality assessments by Tetrastichus planipennisi (Hymenoptera: Eulophidae)

    Science.gov (United States)

    Michael D. Ulyshen; Richard W. Mankin; Yigen Chen; Jian J. Duan; Therese M. Poland; Leah S. Bauer

    2011-01-01

    The biological control agent Tetrastichus planipennisi Yang (Hymenoptera: Eulophidae) is a gregarious larval endoparasitoid of the emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), an invasive cambium-feeding species responsible for recent, widespread mortality of ash (Fraxinus spp.) in...

  12. Crystallization and preliminary X-ray analysis of birch-pollen allergen Bet v 1 in complex with a murine monoclonal IgG Fab' fragment

    DEFF Research Database (Denmark)

    Spangfort, M D; Mirza, Osman Asghar; Gajhede, M

    1999-01-01

    of the clinical symptoms of allergy. In order to study the structural basis of allergen-antibody interaction, a complex between the major birch-pollen allergen Bet v 1 and a Fab' fragment isolated from the murine monoclonal Bet v 1 antibody BV16 has been crystallized. Complex crystals belong to space group P1...

  13. The experimental study on the radioimmunotherapy of the hepatoma in nude mice model with intratumoral injection of 131I-human anti-HBsAg Fab

    International Nuclear Information System (INIS)

    Luo Rongcheng; Wu Guichen; Han Huanxing; You Changxuan; Ding Xuemei; Li Aimin; Wang Chuanbin; Zhang Mingjiang

    2001-01-01

    Objective: To study the therapeutic efficacy of radioimmunotherapy of 131 I-human anti-HBsAg Fab via different routes of administration. Methods: The human hepatoma bearing nude mice we reinjected with 131 I-human anti-HBsAg Fab intra-tumor (IT) and intra-peritoneum (IP). Biodistribution was measured on the 5th day. The tumor growth inhibition rate was determined by measurement of tumor volume. Results: In the IT-treated mice, tumor uptake of 131 I-human anti-HBsAg Fab was four-fold greater than in the IP-treated mice, and normal organ uptake was half of that in the IP-treated mice. At the 3rd week after the infusion, the tumor growth inhibition rate in IT-treated mice was higher than that in the IP-treated mice. Conclusions: Intratumoral administration of 131 I-human anti-HBsAg Fab makes high level of radioactivity retained in tumor with significantly lower radioactivity retained in normal tissues, and provides a more effective regional therapy

  14. Mapping the Binding Interface of VEGF and a Monoclonal Antibody Fab-1 Fragment with Fast Photochemical Oxidation of Proteins (FPOP) and Mass Spectrometry

    Science.gov (United States)

    Zhang, Ying; Wecksler, Aaron T.; Molina, Patricia; Deperalta, Galahad; Gross, Michael L.

    2017-05-01

    We previously analyzed the Fab-1:VEGF (vascular endothelial growth factor) system described in this work, with both native top-down mass spectrometry and bottom-up mass spectrometry (carboxyl-group or GEE footprinting) techniques. This work continues bottom-up mass spectrometry analysis using a fast photochemical oxidation of proteins (FPOP) platform to map the solution binding interface of VEGF and a fragment antigen binding region of an antibody (Fab-1). In this study, we use FPOP to compare the changes in solvent accessibility by quantitating the extent of oxidative modification in the unbound versus bound states. Determining the changes in solvent accessibility enables the inference of the protein binding sites (epitope and paratopes) and a comparison to the previously published Fab-1:VEGF crystal structure, adding to the top-down and bottom-up data. Using this method, we investigated peptide-level and residue-level changes in solvent accessibility between the unbound proteins and bound complex. Mapping these data onto the Fab-1:VEGF crystal structure enabled successful characterization of both the binding region and regions of remote conformation changes. These data, coupled with our previous higher order structure (HOS) studies, demonstrate the value of a comprehensive toolbox of methods for identifying the putative epitopes and paratopes for biotherapeutic antibodies.

  15. Kinetics and tissue distribution of the radiolabeled chimeric monoclonal antibody MOv18 IgG and F(ab')2 fragments in ovarian carcinoma patients

    NARCIS (Netherlands)

    Buist, M. R.; Kenemans, P.; den Hollander, W.; Vermorken, J. B.; Molthoff, C. J.; Burger, C. W.; Helmerhorst, T. J.; Baak, J. P.; Roos, J. C.

    1993-01-01

    Twenty-four patients suspected of having ovarian carcinoma received i.v. injection with a combination of radiolabeled intact IgG (1 mg) and F(ab')2 fragments (1 mg) of the chimeric monoclonal antibody MOv18, each form labeled with 1.85 MBq 131I or 125I. Laparotomy was performed either 2 or 6 days

  16. Pharmacokinetics and tumor targeting of 131I-labeled F(ab')2 fragments of the chimeric monoclonal antibody G250: preclinical and clinical pilot studies.

    NARCIS (Netherlands)

    Brouwers, A.H.; Mulders, P.F.A.; Oosterwijk, E.; Buijs, W.C.A.M.; Corstens, F.H.M.; Boerman, O.C.; Oyen, W.J.G.

    2004-01-01

    INTRODUCTION: Clinical and animal studies of chimeric monoclonal antibody G250 (moAb cG250) for the targeting of clear-cell renal cell carcinoma (RCC), to date, have been with the intact IgG form. To determine whether F(ab')2 fragments are more suited for radioimmunotherapy (RIT) than intact IgG,

  17. Hybrid IgG4/IgG4 Fc antibodies form upon 'Fab-arm' exchange as demonstrated by SDS-PAGE or size-exclusion chromatography

    NARCIS (Netherlands)

    Rispens, Theo; den Bleker, Tamara H.; Aalberse, Rob C.

    2010-01-01

    Human IgG4 antibodies are dynamic molecules that in vivo exchange half-molecules to become bispecific antibodies. Here we show that IgG4 antibodies and IgG4 Fc fragments similarly exchange resulting in hybrid antibodies (a single Fab + Fc) with a molecular weight of ca. 100 kDa. These antibodies can

  18. A Fab fragment directed against the neural cell adhesion molecule L1 enhances functional recovery after injury of the adult mouse spinal cord.

    Science.gov (United States)

    Loers, Gabriele; Cui, Yi-Fang; Neumaier, Irmgard; Schachner, Melitta; Skerra, Arne

    2014-06-15

    Lack of permissive mechanisms and abundance of inhibitory molecules in the lesioned central nervous system of adult mammals contribute to the failure of functional recovery, which leads to severe disabilities in motor functions or pain. Previous studies have indicated that the neural cell adhesion molecule L1 constitutes a viable target to promote regeneration. In the present study, we describe the cloning, functional expression in Escherichia coli cells and purification of a recombinant αL1 Fab fragment that binds to L1 with comparable activity as the function-triggering monoclonal antibody 557.B6 and induces neurite outgrowth and neuronal survival in cultured neurons, despite its monovalent function. Infusion of αL1 Fab into the lesioned spinal cord of mice enhanced functional recovery after thoracic spinal cord compression injury. αL1 Fab treatment resulted in reduced scar volume, enhanced number of tyrosine hydroxylase-positive axons and increased linear density of VGLUT1 (vesicular glutamate transporter 1) on motoneurons. Furthermore, the number and soma size of ChAT (choline acetyltransferase)-positive motoneurons and the linear density of ChAT-positive boutons on motoneurons as well as parvalbumin-positive interneurons in the lumbar spinal cord were elevated. Stimulation of endogenous L1 by application of the αL1 Fab opens new avenues for recombinant antibody technology, offering prospects for therapeutic applications after traumatic nervous system lesions.

  19. Therapeutic IgG4 antibodies engage in Fab-arm exchange with endogenous human IgG4 in vivo

    NARCIS (Netherlands)

    Labrijn, Aran F.; Buijsse, Antonio Ortiz; van den Bremer, Ewald T. J.; Verwilligen, Annemiek Y. W.; Bleeker, Wim K.; Thorpe, Susan J.; Killestein, Joep; Polman, Chris H.; Aalberse, Rob C.; Schuurman, Janine; van de Winkel, Jan G. J.; Parren, Paul W. H. I.

    Two humanized IgG4 antibodies, natalizumab and gemtuzumab, are approved for human use, and several others, like TGN1412, are or have been in clinical development. Although IgG4 antibodies can dynamically exchange half-molecules(1), Fab-arm exchange with therapeutic antibodies has not been

  20. Therapeutic IgG4 antibodies engage in Fab-arm exchange with endogenous human IgG4 in vivo

    NARCIS (Netherlands)

    Labrijn, Aran F.; Buijsse, Antonio Ortiz; van den Bremer, Ewald T. J.; Verwilligen, Annemiek Y. W.; Bleeker, Wim K.; Thorpe, Susan J.; Killestein, Joep; Polman, Chris H.; Aalberse, Rob C.; Schuurman, Janine; van de Winkel, Jan G. J.; Parren, Paul W. H. I.

    2009-01-01

    Two humanized IgG4 antibodies, natalizumab and gemtuzumab, are approved for human use, and several others, like TGN1412, are or have been in clinical development. Although IgG4 antibodies can dynamically exchange half-molecules, Fab-arm exchange with therapeutic antibodies has not been demonstrated

  1. Identification of a conserved protein involved in anaerobic unsaturated fatty acid synthesis in Neiserria gonorrhoeae: implications for facultative and obligate anaerobes that lack FabA

    Science.gov (United States)

    Isabella, Vincent M.; Clark, Virginia L.

    2011-01-01

    SUMMARY Transcriptome analysis of the facultative anaerobe, Neisseria gonorrhoeae, revealed that many genes of unknown function were induced under anaerobic conditions. Mutation of one such gene, NGO1024, encoding a protein belonging to the 2-nitropropane dioxygenase-like superfamiliy of proteins, was found to result in an inability of gonococci to grow anaerobically. Anaerobic growth of an NG1024 mutant was restored upon supplementation with unsaturated fatty acids (UFA), but not with the saturated fatty acid palmitate. Gonococcal fatty acid profiles confirmed that NGO1024 was involved in UFA synthesis anaerobically, but not aerobically, demonstrating that gonococci contain two distinct pathways for the production of UFAs, with a yet unidentified aerobic mechanism, and an anaerobic mechanism involving NGO1024. Expression of genes involved in classical anaerobic UFA synthesis, fabA, fabM, and fabB, was toxic in gonococci and unable to complement a NGO1024 mutation, suggesting that the chemistry involved in gonococcal anaerobic UFA synthesis is distinct from that of the classical pathway. NGO1024 homologs, which we suggest naming UfaA, form a distinct lineage within the 2-nitropropane dioxygenase-like superfamily, and are found in many facultative and obligate anaerobes that produce UFAs but lack fabA, suggesting that UfaA is part of a widespread pathway involved in UFA synthesis. PMID:21895795

  2. A comparison of 111In- or 64Cu-DOTA-trastuzumab Fab fragments for imaging subcutaneous HER2-positive tumor xenografts in athymic mice using microSPECT/CT or microPET/CT

    Science.gov (United States)

    2011-01-01

    Background Our objective was to compare 111In- or 64Cu-DOTA-trastuzumab Fab fragments for imaging small or large s.c. tumor xenografts in athymic mice that display a wide range of human epidermal growth factor receptor-2 (HER2) expression using microSPECT/CT or microPET/CT. Methods Trastuzumab Fab were labeled with 111In or 64Cu by conjugation to 1,4,7,10-tetraazacyclododecane N, N', N'', N'''-tetraacetic acid (DOTA). The purity of 111In- and 64Cu-DOTA-trastuzumab Fab was measured by SDS-PAGE and HPLC. HER2 binding affinity was determined in saturation radioligand binding assays using SKBR-3 cells (1.3 × 106 HER2/cell). MicroSPECT/CT and microPET/CT were performed in athymic mice bearing s.c. BT-20 and MDA-MB-231 xenografts with low (0.5 to 1.6 × 105 receptors/cell), MDA-MB-361 tumors with intermediate (5.1 × 105 receptors/cell) or SKOV-3 xenografts with high HER2 expression (1.2 × 106 receptors/cell) at 24 h p.i. of 70 MBq (10 μg) of 111In-DOTA-trastuzumab Fab or 22 MBq (10 μg) of 64Cu-DOTA-trastuzumab Fab or irrelevant 111In- or 64Cu-DOTA-rituximab Fab. Tumor and normal tissue uptake were quantified in biodistribution studies. Results 111In- and 64Cu-DOTA-trastuzumab were > 98% radiochemically pure and bound HER2 with high affinity (Kd = 20.4 ± 2.5 nM and 40.8 ± 3.5 nM, respectively). MDA-MB-361 and SKOV-3 tumors were most clearly imaged using 111In- and 64Cu-DOTA-trastuzumab Fab. Significantly higher tumor/blood (T/B) ratios were found for 111In-DOTA-trastuzumab Fab than 111In-DOTA-rituximab Fab for BT-20, MDA-MB-231 and MDA-MB-361 xenografts, and there was a direct association between T/B ratios and HER2 expression. In contrast, tumor uptake of 64Cu-DOTA-trastuzumab Fab was significantly higher than 64Cu-DOTA-rituximab Fab in MDA-MB-361 tumors but no direct association with HER2 expression was found. Both 111In- and 64Cu-DOTA-trastuzumab Fab imaged small (5 to 10 mm) or larger (10 to 15 mm) MDA-MB-361 tumors. Higher blood, liver, and spleen

  3. Vertical Distribution and Daily Flight Periodicity of Ambrosia Beetles (Coleoptera: Curculionidae) in Florida Avocado Orchards Affected by Laurel Wilt.

    Science.gov (United States)

    Menocal, Octavio; Kendra, Paul E; Montgomery, Wayne S; Crane, Jonathan H; Carrillo, Daniel

    2018-03-08

    Ambrosia beetles have emerged as significant pests of avocado ((Persea americana Mill. [Laurales: Lauraceae])) due to their association with pathogenic fungal symbionts, most notably Raffaelea lauricola T.C. Harr., Fraedrich & Aghayeva (Ophiostomatales: Ophiostomataceae), the causal agent of the laurel wilt (LW) disease. We evaluated the interaction of ambrosia beetles with host avocado trees by documenting their flight height and daily flight periodicity in Florida orchards with LW. Flight height was assessed passively in three avocado orchards by using ladder-like arrays of unbaited sticky traps arranged at three levels (low: 0-2 m; middle: 2-4 m; high: 4-6 m). In total, 1,306 individuals of 12 Scolytinae species were intercepted, but six accounted for ~95% of the captures: Xyleborus volvulus (Fabricius) (Coleoptera: Curculionidae), Xyleborinus saxesenii Ratzeburg (Coleoptera: Curculionidae), Euplatypus parallelus (Fabricius) (Coleoptera: Curculionidae), Xyleborus bispinatus Eichhoff (Coleoptera: Curculionidae), Xyleborus affinis Eichhoff (Coleoptera: Curculionidae), and Hypothenemus sp. (Coleoptera: Curculionidae). The primary vector of R. lauricola, Xyleborus glabratus Eichhoff (Coleoptera: Curculionidae), was not detected. Females of X. volvulus showed a preference for flight at low levels and X. bispinatus for the low and middle levels; however, captures of all other species were comparable at all heights. At a fourth orchard, a baiting method was used to document flight periodicity. Females of X. saxesenii and Hypothenemus sp. were observed in flight 2-2.5 h prior to sunset; X. bispinatus, X. volvulus, and X. affinis initiated flight at ~1 h before sunset and Xylosandrus crassiusculus (Motschulsky) (Coleoptera: Curculionidae) at 30 min prior to sunset. Results suggest that ambrosia beetles in South Florida fly near sunset (when light intensity and wind speed decrease) at much greater heights than previously assumed and have species-specific patterns in host

  4. Bioacoustics of Acanthoscelides obtectus (Coleoptera: Chrysomelidae: Bruchinae) on Phaseolus vulgaris (Fabaceae)

    Science.gov (United States)

    Acanthoscelides obtectus (Say) (Coleoptera: Chrysomelidae), is an economically important pest of common bean Phaseolus vulgaris L. (Fabaceae) in the tropics and subtropics. It is difficult to detect the presence of A. obtectus because the larvae are cryptic and spend most of their developmental time...

  5. Hydraena Kugelann, 1794 (Coleoptera, Hydraenidae from the Seychelles, Indian Ocean, with description of a new species

    Directory of Open Access Journals (Sweden)

    Manfred A. Jäch

    2016-10-01

    Full Text Available Hydraena matyoti sp. n. (Coleoptera, Hydraenidae is described from the Seychelles, Indian Ocean. Hydraena mahensis Scott, 1913 is redescribed. The latter is here recorded from La Digue for the first time. A key to the species of the genus Hydraena Kugelann, 1794 of the Seychelles is presented.

  6. Hydraena Kugelann, 1794 (Coleoptera, Hydraenidae) from the Seychelles, Indian Ocean, with description of a new species.

    Science.gov (United States)

    Jäch, Manfred A; Delgado, Juan A

    2016-01-01

    Hydraena matyoti sp. n. (Coleoptera, Hydraenidae) is described from the Seychelles, Indian Ocean. Hydraena mahensis Scott, 1913 is redescribed. The latter is here recorded from La Digue for the first time. A key to the species of the genus Hydraena Kugelann, 1794 of the Seychelles is presented.

  7. Review of the genus Ceresium Newman, 1842 (Coleoptera: Cerambycidae) in Fiji

    Science.gov (United States)

    A taxonomic review of the genus Ceresium (Coleoptera: Cerambycidae) found within the Fiji Islands is presented. A total of 17 species is treated. Full morphological descriptions and comparative images of each species are included, along with a dichotomous key for their identification....

  8. Review of the genus Ceresium Newman, 1842 (Coleoptera, Cerambycidae) in Fiji

    Science.gov (United States)

    Waqa-Sakiti, Hilda; Winder, Linton; Lingafelter, Steven W.

    2015-01-01

    Abstract A taxonomic review of the genus Ceresium (Coleoptera: Cerambycidae) found within the Fiji Islands is presented. A total of 17 species is treated. Full morphological descriptions and comparative images of each species are included, along with a dichotomous key for their identification. PMID:26692805

  9. Use of nutrient self selection as a diet refining tool in Tenebrio molitor (Coleoptera: Tenebrionidae)

    Science.gov (United States)

    A new method to refine existing dietary supplements for improving production of the yellow mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae), was tested. Self selected ratios of 6 dietary ingredients by T. molitor larvae were used to produce a dietary supplement. This supplement was compared...

  10. Impact of Adult Weight, Density, and Age on Reproduction of Tenebrio molitor (Coleoptera: Tenebrionidae)

    Science.gov (United States)

    The impact of adult weight, age, and density on reproduction of Tenebrio molitor L. (Coleoptera: Tenebrionidae) was studied. The impact of adult weight on reproduction was determined in two ways: 1) counting the daily progeny of individual adult pairs of known weight and analyzing the data with line...

  11. Temperature influences on diapause induction and survival in the boll weevil (Coleoptera: Curculionidae)

    Science.gov (United States)

    The boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), has been the most important pest of cotton (Gossypium spp.) wherever it occurs. Although eradication programs in the U.S. have reduced the range of this pest, the weevil remains an intractable problem in subtropical Tex...

  12. Development of an improved attractive lure for the pine shoot beetle, Tomicus piniperda (Coleoptera: Scolytidae)

    Science.gov (United States)

    Therese M. Poland; Peter de Groot; Stephen Burke; David Wakarchuk; Robert A. Haack; Reginald Nott; Taylor Scarr

    2003-01-01

    1) The pine shoot beetle, Tomicus piniperda (L.) (Coleoptera: Scolytidae), is an exotic pest of pine, Pinus spp., and was first discovered in North America in 1992. 2) Although primary attraction to host volatiles has been clearly demonstrated for T. piniperda, the existence and role of secondary attraction to...

  13. Influence of temperature on spring flight initiation for southwestern ponderosa pine bark beetles (Coleoptera: Curculionidae, Scolytinae)

    Science.gov (United States)

    M. L. Gaylord; K. K. Williams; R. W. Hofstetter; J. D. McMillin; T. E. Degomez; M. R. Wagner

    2008-01-01

    Determination of temperature requirements for many economically important insects is a cornerstone of pest management. For bark beetles (Coleoptera: Curculionidae, Scolytinae), this information can facilitate timing of management strategies. Our goals were to determine temperature predictors for flight initiation of three species of Ips bark beetles...

  14. Tanyproctus (Tanyproctus) arher (Coleoptera: Scarabaeidae: Melolonthinae: Tanyproctini), a new species from the Socotra Island, Yemen

    Czech Academy of Sciences Publication Activity Database

    Bezděk, Aleš; Sehnal, R.; Král, D.

    2013-01-01

    Roč. 3737, č. 2 (2013), s. 191-196 ISSN 1175-5326 Institutional support: RVO:60077344 Keywords : Coleoptera * Scarabaeoidea * Scarabaeidae Subject RIV: EG - Zoology Impact factor: 1.060, year: 2013 http://www.mapress.com/zootaxa/2013/f/zt03737p196.pdf

  15. Stomanomala subcostata (Coleoptera: Scarabaeidae), the first record of ruteline chafer from Socotra Archipelago, Yemen

    Czech Academy of Sciences Publication Activity Database

    Bezděk, Aleš; Král, D.; Limbourg, P.

    2017-01-01

    Roč. 57, č. 3 (2017), s. 87-91 ISSN 0374-1036 Institutional support: RVO:60077344 Keywords : Coleoptera * Scarabaeidae * Rutelinae Subject RIV: EG - Zoology OBOR OECD: Zoology Impact factor: 0.632, year: 2016 https://www.biotaxa.org/AEMNP/article/view/35053

  16. Microbial control of Agrilus planipennis (Coleoptera: Buprestidae) with Beauveria bassiana strain GHA: field applications

    Science.gov (United States)

    Houping Liu; Leah S. Bauer

    2008-01-01

    The effects of Beauveria bassiana strain GHA, applied as BotaniGard ES, on newly colonised and well-established populations of emerald ash borer, Agrilus planipennis (Coleoptera: Buprestidae) were evaluated in the field using foliar and trunk sprays in Michigan in 2004-2005. Results from field trials at a newly colonised white ash...

  17. Study on the genus Daptus ground-beetles (Coleoptera: Carabidae from Korea

    Directory of Open Access Journals (Sweden)

    Ik Je Choi

    2016-03-01

    Full Text Available A genus Daptus Fischer von Waldheim, 1823 of the tribe Harpalini Bonelli, 1810 (Coleoptera: Carabidae is reported for the first time from Korea, based on the Daptus vittatus Fischer von Waldheim from Incheon, Korea. Redescription of the species and illustrations of diagnostic characteristics, including genitalia characteristics of both sexes, are provided.

  18. Molecular Diagnostic for Boll Weevil (Coleoptera: Curculionidae) Based on Amplification of Three Species-specific Microsatellites

    Science.gov (United States)

    The boll weevil, Anthonomus grandis Boheman (Coleoptera: Curculionidae), is a serious pest of cultivated cotton in the Americas, and reinfestation of zones from which they have been eradicated is of perpetual concern. Extensive arrays of pheromone traps monitor for reintroductions, but occasionally...

  19. Flight propensty of Anoplophora glabripennis, an Asian longhorned beetle (Coleoptera: Cerambycidae)

    Science.gov (United States)

    J. A. Francese; B. Wang; D. R. Lance; Z. Xu; S. Zong; Y. Luo; A. J. Sawyer; V. C. Mastro

    2003-01-01

    Anoplophora glabripennis (Coleoptera: Cerambycidae) (Motschulsky), is a recently introduced pest of hardwoods. Research to study its flight behavior was conducted in the field in Ningxia Autonomous Region, Peoples' Republic of China. To study the flight propensity of A. glabripennis, adult beetles were observed in population...

  20. Characterization of an Aggregation Pheromone in Hylesinus pruinosus (Coleoptera: Curculionidae: Scolytinae)

    Science.gov (United States)

    William Shepherd; Brian Sullivan; Bradley Hoosier; JoAnne Barrett; Tessa Bauman

    2010-01-01

    We conducted laboratory and field bioassays to characterize the pheromone system of an ash bark beetle, Hylesinus pruinosus Eichhoff (Coleoptera: Curculionidae: Scolytinae). Solitary females in newly initiated galleries in ash logs produced (+)-exo-brevicomin, whereas male beetles paired with females produced (+)-endo-brevicomin, lesser quantities of...