Approximately 30% of foxes given two doses of an inactivated rabies antigen delivered directly into the intestinal tract developed an immune response as measured by rabies serum neutralizing antibodies....Full Text Available
The efficacy of a heat inactivated hepatitis B virus vaccine, containing 3 micrograms hepatitis B surface antigen (HBsAg), was studied in a high risk group of 800 susceptible homosexual men by a randomised...Full Text Available
Israel has faced the challenge presented by epidemic poliomyelitis by using different immunization strategies. In the 1950s, inactivated poliovirus vaccine (IPV) helped to reduce the total burden of...Full Text Available
BackgroundAntigen sparing and cross-protective immunity are regarded as crucial in pandemic influenza vaccine development. Both targets can be achieved by adjuvantation strategy...Full Text Available
Hepatitis B subviral particles, purified from plasma of asymptomatic carriers seropositive for hepatitis B e antigen, were treated with various conditions reported for the processing of vaccines. Thereafter,...Full Text Available
Data from 52 litters farrowed in two seasons of a cross-fostering experiment were analyzed to estimate variances and covariances for additive direct and maternal genetic effects on immune response to pseudorabies virus and B. bronchiseptica vaccine. Twenty purebred boars and 44 sows of the Duroc, Landrace and Yorkshire breeds were used. Immune response was measured after vaccine challenge. A modified-live pseudorabies (PR) vaccine was administered to piglets at 28 d of age; response was measured by log2 serum neutralization titers at 56 d. An inactivated B. bronchiseptica bacterin was administered at 28, 42 and 112 d. Antibody levels were measured relative to positive and negative controls at 28, 56 and 119 d by using an enzyme-linked immunosorbent assay (ELISA). Results from this study for heritability were .18 +/- .09 for PR titer and .15 +/- .07 and .52 +/- .15 for 56- and 119-d ELISA values, ...
Radiation inactivation and sedimentation equilibrium analysis were used to determine the functional and physical size of the chicken hepatic membrane receptor that binds N-acetylglucosamine-terminated glycoproteins. Purified plasma membranes from chicken liver were irradiated with high energy electrons and assayed for 125I-agalactoorosomucoid binding. Increasing the dose of ionizing radiation resulted in a monoexponential decay in binding activity due to a progressive loss of binding sites. The molecular mass of the chicken lectin, determined in situ by target analysis, was 69,000 +/- 9,000 Da. When the same irradiated membranes were solubilized in Brij 58 and assayed, the binding protein exhibited a target size of 62,000 +/- 4,000 Da; in Triton X-100, the functional size of the receptor was 85,000 +/- 10,000 Da. Sedimentation equilibrium measurements of the purified binding protein yielded a lower limit molecular weight of ...
... a flu shot There is a new nasal spray version of the vaccine, and it contains weakened ... the LAIV (Live Attenuated Influenza Vaccine). The nasal-spray flu vaccine contains three different live but weakened ...
home when you are sick helps everyone you care about Questions & Answers Flu Vaccine: Q&A Vaccine Dosage & Administration Vaccine Supply for 2011-12 Season Vaccine Supply and...
home when you are sick helps everyone you care about Questions & Answers Flu Vaccine: Q&A Vaccine Dosage & Administration Vaccine Supply for 2011-12 Season Vaccine Supply and...
home when you are sick helps everyone you care about Questions & Answers Flu Vaccine: Q&A Vaccine Dosage & Administration Vaccine Supply for 2011-12 Season Vaccine Supply and...
Ten children receiving maintenance dialysis were immunized with the standard dose of measles-mumps-rubella vaccine between 15 and 33 months of age. Immune responses to vaccination were determined using commercially available enzyme-linked immunosorbent assays for measles, mumps, and rubella viruses. Eight children responded to measles vaccine, 5 to mumps vaccine, 8 to rubella vaccine, and only 3 children to all three vaccines, compared with a seroconversion rate of over 90% to all three vaccines in healthy children (P less than 0.0001). We contend that the relatively poor immunocompetence of our dialysis patients explains their less than optimal vaccine response and suggest that children vaccinated while undergoing dialysis be tested to confirm serological evidence of immunity. PMID:1571219
The first candidate rotavirus vaccine was a live attenuated oral vaccine made by the classical empirical method of serial passage of virus in tissue culture cells. Current tetravalent vaccine candidates that are in the final stages of efficacy testing in the United States were made by genetic reassortment. This article briefly highlights how advances in the basic understanding of the molecular biology of rotaviruses have facilitated vaccine development. New approaches for second-generation vaccines and improvements in vaccine efficacy based on further exploitation of the tools and knowledge of rotavirus molecular biology and pathogenesis are discussed. PMID:8752289
Ninety percent of foxes fed commercial ERA vaccine in a specially designed bait developed rabies serum neutralizing antibodies. The vaccine bait did not cause clinical signs of rabies when consumed...Full Text Available
BACKGROUND: Although the effectiveness of influenza vaccination in high-risk groups has been proven, vaccine coverage continues to be less than 50% in The Netherlands. To improve vaccination rates,...Full Text Available
SummaryBecause increasing numbers of HIV vaccine candidates are being tested globally, it is essential to differentiate vaccine-from virus-induced antibodies. Most of the currently...Full Text Available
The extensive diversity of Plasmodium falciparum antigens is a major obstacle to a broadly effective malaria vaccine but population genetics has rarely been used to guide vaccine design....Full Text Available
BackgroundThe first vaccine to prevent human papillomavirus (HPV) and cervical cancer has been licensed, and in future, vaccination may be routinely offered to 10–14 year...Full Text Available
The mechanism of trypsin inactivation by intact Hymenolepis diminuta has been investigated by biochemical and autoradiographic methods. Although worms inactivate trypsin and chymotrypsin in vitro, no inactivation of other endoproteases (subtilisin, pepsin and papain) could be demonstrated. Trypsin inactivation, as demonstrated by macromolecular substrates (azoalbumin, hemoglobin and casein), could not be detected using low molecular weight synthetic substrates such as N-p-benzoyl-DL-arginine-p-nitroanilide (BAPA) or N-p-tosyl-L-arginine methyl ester (TAME). In addition, the kinetic parameters (K/sub m/ and k_3) for H. diminuta-inactivated trypsin, using BAPA as the substrate, were not different from those of the native enzyme. The number of active sites for both native and inactivated trypsin were determined by titration with p-nitro-phenyl-p'-guanidinobenzoate. ...
The evolution of "humanized" (i.e., free of animal sourced reagents) and ultimately chemically defined culture systems for human embryo stem cell (hESC) isolation and culture is of importance to improving their efficacy and safety in research and therapeutic applications. This can be achieved by integration of a multitude of individual approaches to replace or eliminate specific animal sourced reagents into a single comprehensive protocol. In the present study our objective was to integrate strategies obviating reliance on some of the most poorly defined and path-critical factors associated with hESC derivation, namely the use of animal immune compliment to isolate embryo inner cell mass, and animal sourced serum products and feeder cells to sustain hESC growth and attachment. As a result we report the derivation of six new hESC lines isolated by outgrowth from whole blastocysts on an extracellular matrix substrate of purified human laminin (Ln) with transitional ...
The photoaffinity probes [#gamma#-"3"2P]2-azidoATP (2-N_3ATP) and [#alpha#-"3"2P]8-azido-ATP (8-N_3ATP) were used to investigate the binding of ATP to highly purified 2-5A synthetase. 2-N_3APT and 8-N_3ATP are substrates for 2-5A synthetase. In this study the authors show that 2- and 8-N_3ATP are competitive inhibitors of the enzymatic conversion of ATP to 2-5A. Ultraviolet irradiation results in the photoinsertion of 2-N_3ATP and 8-N_3ATP into the enzyme. The covalent photoinsertion of [#alpha#-"3"2P]8-N_3ATP into the 2-5A synthetase is proportional to the inactivation of the enzyme as UV irradiation is increased. Photolabeling of 2-5A synthetase is saturated at 1.5 mM 2-N_3ATP and 2.0 mM 8-N_3ATP. Computer analysis of the curvilinear Scatchard plots of the 2-5A synthetase suggest the presence of high-affinity and low-affinity binding sites that may correspond to the acceptor and the 2'-adenylation sites of the enzyme. The competition of ...
Nitrate reductase from wheat (Triticum aestivum L. cv Bindawarra) leaves is inactivated by pretreatment with NADH, in the absence of nitrate, a 50% loss of activity occurring in 30...Full Text Available
Phenothiazine-N-carbonyl chloride inactivated chymotrypsin and trypsin by means of a 1:1 stoicheiometric reaction. Its reaction with chymotrypsin was 29 times as fast as that with...Full Text Available
Sustainable management of toilet waste must prevent disease transmission but allow reuse of plant nutrients. Inactivation of uterus-derived Ascaris suum eggs was studied in relation...Full Text Available
Treatment of poliovirus Type I with 10−3m 2-thiouracil (2-TU) resulted in the inactivation of greater than 90% of the virus infectivity and stabilization of approximately...Full Text Available
This work investigated the inactivation of Microcystis aeruginosa (MA) with contact glow discharge electrolysis (CGDE). The influences of applied voltage, current and treatment time on the inactivation rate were critically examined. Based on the above results, the optimal conditions were chosen to sufficiently utilize chemically active species and enhance the inactivation of MA. Under the optimal conditions (voltage: 530?V; current: 30?mA; treatment time: 20?min), the inactivation rate of algae was more than 90% within 5?days incubation after inoculating. At the same time, the concentrations of Chlorophyll-a and dehydrogenase decreased, which demonstrated that 20?min CGDE treatment could effectively inhibit the growth of MA and caused deterioration of cell integrity. The present work would...
The invention relates to a method for purifying a gas mixture containing at least one undesirable gas compound utilizing certain sulfonamide or sulfamide solvents.
There is a quest for the development of a new generation of vaccines consisting of well-defined subunit antigens. For a number of practical reasons it is attractive to develop vaccines on the basis...Full Text Available
The human immunodeficiency virus Tat protein is essential for virus replication and is a candidate vaccine antigen. Macaques immunized with Tat or chemically modified Tat toxoid having the same clade...Full Text Available
The development of an HIV vaccine will require a more precise understanding of the immunological and virological underpinnings of HIV infection. Magnetofection, the process of magnetizing HIV...Full Text Available
Although major inroads into making antiretroviral therapy available in resource-poor countries have been made, there is an urgent need for an effective vaccine administered shortly after birth, which...Full Text Available
... Weiss, S., D. Kobiler, H. Levy, H. Marcus, A. Pass, N. Rothschild, and Z ... of Bacillus anthracis spores conferred by a protective antigen-based vaccine in rabbits ...
The attitude of Nigerian mothers, mainly Yoruba, towards measles vaccine and other aspects of prevention are influenced by different perceptions of the cause of measles. There is a significant correlation...Full Text Available
The entire genome of rabies virus vaccine strain Flury-LEP-C, a Chinese variant of the rabies virus vaccine strain Flury-LEP, was sequenced. The overall length of the genome of Flury-LEP-C strain was...Full Text Available
BackgroundAlthough the World Health Organization had recommended that every child be vaccinated for Hepatitis B by the early 1980s, large multinational pharmaceutical companies held...Full Text Available
During the anamnestic response, the sera of four rabbits immunized with a polyvalent pneumococcal vaccine contained large amounts of cryoglobulins belonging to the G and M immunoglobulin classes. These...Full Text Available
Splenectomy or pretreatment of adult hamsters with cyclophosphamide (Cytoxan) increased the lethality of the TC-83 vaccine strain of Venezuelan encephalitis virus (VEE), inoculated subcutaneously, from 12% for normal hamsters to 75 and 76% respectively. A...
PurposeWe previously reported a randomized phase II clinical trial combining a poxvirus-based vaccine encoding PSA with radiotherapy in patients with localized prostate...Full Text Available
The effect of abatacept, a selective T-cell co-stimulation modulator, on vaccination has not been previously investigated. In this open-label, single-dose, randomized, parallel-group, controlled study,...Full Text Available
We studied the effect of booster injections and the long-term immune response after injections of an anti-human immunodeficiency virus type 1 (HIV-1) lipopeptide vaccine. This vaccine was injected alone...Full Text Available
A field study was conducted under commercial feedlot conditions at 2 sites in western Canada to determine the relative effects of a univalent viral vaccine (MLV 1) program versus a multivalent viral...Full Text Available
Introduction: Tumor-derived heat shock protein (HSP)-peptide complexes (HSPPCs) induced immunity against malignancies in preclinical trials, working across tumor types and bypassing the need to identify single immunogenic peptides. These results paved the way for the use of human gp96 obtained from autologous tumor samples as an anti-cancer vaccine. Areas covered: Autologous tumor-derived HSP gp96 peptide complex (HSPPC-96) vaccine is emerging as a tumor- and patient-specific cancer vaccine, with confirmed activity in several malignancies. It has been tested in Phase III clinical trials in advanced melanoma and kidney cancer with evidence for efficacy in patients with earlier stage disease. HSPPC-96-based vaccine demonstrated an excellent safety profile, thus emerging as a novel therapeuti...
Inactivation of aflatoxin B1 was studied by using gamma radiation and hydrogen peroxide. A 100-krad dose of gamma radiation was sufficient to inactivate 50 micrograms of aflatoxin B1 in the presence of 5% hydrogen peroxide, and 400 krad was required for total degradation of 100 micrograms of aflatoxin in the same system. Degradation of aflatoxin B1 was confirmed by high-pressure liquid chromatographic and thin-layer chromatographic analysis. Ames microsomal mutagenicity test showed loss of aflatoxin activity. This method of detoxification also reduces the toxin levels effectively in artificially contaminated groundnuts.
Eight different viruses, suspended in drinking water, were examined for their ability to be inactivated by microwaves from a microwave oven. Up to a virus content of 10/sup 5/ TCID/sub 50//ml inactivation was successful within a few minutes of microwave treatment and occurred in parallel to the heat stability of the viruses. Evidence for direct effects of microwaves on viruses could not be detected. 7 of the viruses studied were inactivated rapidly when temperatures of 50 to 65/sup 0/C under microwave treatment were reached in the flowing water, while a bovine parvovirus was only inactivated by temperatures above 90/sup 0/C. The advantages of a thermal virus-decontamination of fluids and material by microwaves are discussed.
The Molecular Genetics and Carcinogenesis Section conducts studies using human epithelial cells to assess: activation of proto-oncogenes by chemical and physical carcinogens; inactivation and dysregulation of tumor suppressor genes by chemical and physical
A new semi-empirical algorithm for the radial distribution of dose is compared with available data. The algorithm is used to calculate the inactivation cross section for dry enzymes and viruses using an extended target model of a 1-hit detector. Agreement with data is at about the 15% level, approximating the precision of the data itself. (author).
The mechanism of inactivation of cytochrome P450 2B1 (CYP2B1) by 4-tert-butylphenylacetylene (BPA) has been characterized previously to be caused by the covalent binding of a reactive...Full Text Available
Activities of glycogen synthase (GS) and GS phosphatase were determined on human muscle biopsies before and after isometric contraction at 2/3 maximal voluntary force. Total GS activity did not change...Full Text Available
Persistent hepatitis C virus (HCV) infection affects 170 million people worldwide. Acute HCV infection is often asymptomatic, but many infected individuals develop persistent infections that may lead to development of end-stage liver diseases, including liver cirrhosis and hepatocellular carcinoma. Thus, an HCV vaccine that could significantly lower the chronicity rate would have a major impact on the disease burden. Unfortunately, HCV is a highly mutable virus, and escape mutations can undermine vaccine-induced virus-specific immunity. Also, HCV exists as multiple genotypes, and so genotype-specific vaccines might be required to achieve broad protection. Finally, vaccine development has been hampered by the lack of a small animal model and cell culture systems, but these are currently being established. Despite these obstacles, several vaccine candidates tested in the chimpanzee ...
The clotting activity of Staphylococcus aureus strain 104 was purified 46,000-fold, but absolute purity was not achieved. Carbohydrate content of the purified material was not more than 5%. Elution...Full Text Available
Isolated cauliflower (Brassica oleracea) bud plastids, purified by isopycnic centrifugation in density gradients of Percoll, were found to be highly intact, to be practically devoid...Full Text Available
The aim of the present study was to describe the physicochemical characteristics of streptococcal T antigen. T protein isolated from Streptococcus pyogenes type 12 (R53/1077, Colindale) and purified...Full Text Available
IntroductionThe outbreak of cholera in Zimbabwe intensified interest in the control and prevention of cholera. While there is agreement that safe water, sanitation, and personal...Full Text Available
Age-related alterations in the cellular composition of the B lineage are a major cause of the poor antibody response to vaccination and to infectious agents among the elderly population. The mechanisms...Full Text Available
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A letter of inquiry was sent to 21 countries in Europe to ascertain their experience of diphtheria during the last 20 years. An analysis of the 15 replies showed that all relied on childhood vaccination...Full Text Available
BackgroundAlthough advances in rehydration therapy have made cholera a treatable disease with low case-fatality in settings with appropriate medical care, cholera continues...Full Text Available
... view on the utility of such attacks, see JH Rothschild, Tomorrow's Weapons ... Aum Shinrikyo may have had a stolen vaccine anthrax strain incapable ...
Background and aims. The multiplicity of vaccine injections during childhood leads to iterative painful and stressful experiences which may lead in turn to anticipated pain and then possibly to a true needle phobia. We aimed at evaluating a multifactorial strategy of pain management combining pharmacological and non-pharmacological approaches during vaccination, as compared to usual care, in 4- to 12-year-old children. Methods. In all, 239 children were enroled by 25 family practitioners in an open-label study. After a pseudo-randomization, usual pain management (n = 132) was compared to a multifactorial strategy (n = 107) associating preliminary application of an anesthesic patch, preferential use of specified vaccines, child education by the parents and the doctor, parental accompaniment...
BackgroundMycobacterium avium subspecies paratuberculosis causes systemic infection and chronic intestinal inflammation in many species including...Full Text Available
This study was focused on the possibility to inactivate main food pathogens, their spores and biofilms on the surface of packaging material polyolefine by Na-chlorophyllin (Na-Chl)-based photosensitization and to compare efficiency of this treatment with conventional antimicrobials. Data indicate that Bacillus cereus and Listeria monocytogenes were effectively inactivated (7 log) by Na-Chl (7.5x10-7M)-based photosensitization in vitro and on the surface of packaging. Meanwhile to achieve adequate inactivation of thermo-resistant strains, spores or biofilms the higher Na-Chl concentration and longer illumination times had to be used. Comparison of different surface decontamination treatments reveal that photosensitization is much more effective against B. cereus and L. monocytogenes attache...
This study was focused on the possibility to inactivate thermosensitive Listeria monocytogenes ATCL3C 7644 and thermoresistant 56 Ly strain by Na-Chlorophyllin (Na-Chl)-based photosensitization in vitro and on the surface of packaging. Comparative analysis of antimicrobial efficiency of photosensitization with conventional surface cleaning was performed. Data indicate that both Listeria strains, after incubation with Na-Chl and following illumination (l=400nm, 20mWcm-2), were inactivated by 7 log in vitro. This treatment cleaned both Listeria strains from packaging surfaces. Comparative analysis indicates that washing with water diminishes pathogens by less than 1 log, 200ppm Na-hypochlorite by 1.7 log, Na-Chl-based photosensitization by 4.5 log. Listeria biofilms were totally removed from...
BackgroundThe Plasmodium falciparum pre-erythrocytic stage candidate vaccine RTS,S is being developed for protection of young children against...Full Text Available
Vibrio cholerae 638 (El Tor, Ogawa), a new CTXΦ-negative hemagglutinin/protease-defective strain that is a cholera vaccine candidate, was examined for safety and immunogenicity...Full Text Available
Adenovirus vaccine vectors derived from rare human serotypes have been shown to be less potent than serotype 5 (Ad5) at inducing immune responses to encoded antigens. To identify highly immunogenic...Full Text Available
...INFORMATION Diarrhoeal disease Food safety and foodborne illness Enterohaemorrhagic Escherichia coli (EHEC) Cholera WHO PROGRAMMES AND ACTIVITIES Child and Adolescent Health and Development (...FOS) Initiative for Vaccine Research (IVR) TECHNICAL INFORMATION Vaccine research: diarrhoeal diseases Cholera Water-related diseases Household water treatment and safe storage WHO Global Salm-Surv ...PUBLICATIONS Diarrhoea: child and adolescent health Diarrhoea: cholera RELATED TOPICS - Child health - Water - Food safety - Cholera - Travel - Breastfeeding ...
The use of tick vaccines in mammalian hosts has been shown to be the most promising alternative tick control method to current use of acaricides, which suffers from a number of limitations. However,...Full Text Available
Veterinarians and farmers employing multivalent killed vaccines in lactating dairy cows have reported transient losses in milk production. Few studies have quantified this loss. In this report, effects...Full Text Available
The Yellow Fever Vaccine 17D (YFV17D) has been used as a vector for the Lassa virus glycoprotein precursor (LASV-GPC) resulting in construction of YFV17D/LASV-GPC recombinant virus. The virus...Full Text Available
BackgroundThe malaria vaccine candidate antigen RTS,S includes parts of the pre-erythrocytic stage circumsporozoite protein fused to the Hepatitis B surface antigen. Two Adjuvant...Full Text Available
Pathogenic strains of Vibrio cholerae are lysogens of the filamentous phage CTXφ, which carries the genes for cholera toxin (ctxAB). We found that the titers...Full Text Available
Recessive inactivating mutations in human matrix metalloproteinase 2 (MMP2, gelatinase A) are associated with syndromes that include abnormal facial appearance, short stature, and severe bone...Full Text Available
Staphylococcus aureus preferentially catabolizes glucose, generating pyruvate, which is subsequently oxidized to acetate under aerobic growth conditions. Catabolite repression of the...Full Text Available
The in situ assembly states of the glucose transport carrier protein in the plasma membrane and in the intracellular (microsomal) storage pool of rat adipocytes were assessed by studying radiation-induced inactivation of the D-glucose-sensitive cytochalasin B binding activities. High energy radiation inactivated the glucose-sensitive cytochalasin B binding of each of these membrane preparations by reducing the total number of the binding sites without affecting the dissociation constant. The reduction in total number of binding sites was analyzed as a function of radiation dose based on target theory, from which a radiation-sensitive mass (target size) was calculated. When the plasma membranes of insulin-treated adipocytes were used, a target size of approximately 58,000 daltons was obtained. For adipocyte microsomal membranes, we obtained target sizes of approximately 112,000 and 109,000 daltons prior to and after insulin treatment, ...
Neutral endopeptidase 24.11 (NEP, CD10) is a cell-surface enzyme expressed by prostatic epithelial cells that cleaves and inactivates neuropeptides implicated in the growth of androgen-independent prostate...Full Text Available
Scytalidium thermophilum isolates in culture, as well as the endogenous strain(s) in mushroom compost, were inactivated at 70°C. This temperature was used to pasteurize composts...Full Text Available
Hyperproliferation of the colonic epithelium, leading to expansion of colonic crypt progenitors, is a recognized risk factor for colorectal cancer. Overexpression of progastrin, a nonamidated and incompletely...Full Text Available
Isogenic strains containing insertional disruptions of 10 Haemophilus influenzae Rd genes were investigated for their effects on the susceptibility of the organism to various classes...Full Text Available
Although most self-reactive T cells are eliminated in the thymus, mechanisms to inactivate or control T cells specific for extrathymic antigens are required and exist in the periphery. By investigating...Full Text Available
Inactivation of the RB tumor suppressor and activation of the MYC family of oncogenes are frequent events in a large spectrum of human cancers. Loss of RB function and MYC activation are thought to...Full Text Available
Heat inactivated Aspergillus ustus (Asp), silicon dioxide-nano-powder (N Si), and silicon dioxide nano-powder-combined-heat inactivated Aspergillus ustus (N Si Asp) were used to study the biosorption of Cd(II) from aqueous solutions via batch equilibrium technique. Surface characterization and immobilization of the fungal cells on silicon dioxide-nano-powder were examined and confirmed by using FT-IR and ESM analysis. Cadmium biosorption processes were investigated under the effect of pH, contact time, sorbent dosage and initial metal concentration. The three examined sorbents were found to exhibit maximum mmolg^-^1 capacity values in pH 7.0. The maximum determined cadmium capacity by silicon dioxide-nano-powder (N Si) (600mmolg^-^1) was found higher than that exhibited by the heat inactiv...
Mutational inactivation of the RB1 tumor suppressor gene initiates retinoblastoma and other human cancers. RB1 protein (pRb) restrains cell proliferation by binding...Full Text Available
In order to identify novel high value antibacterial targets it is desirable to delineate whether the inactivation of the target enzyme will lead to bacterial death or stasis. This knowledge is particularly...Full Text Available
The pathogenic mechanisms of human autosomal dominant polycystic kidney disease (ADPKD) have been well known to include the mutational inactivation of PKD2. Although haploinsufficiency...Full Text Available
The most characteristic features of the Lyme disease pathogens, the Borrelia burgdorferi sensu lato (s.l.) group, are their ability to invade tissues and to circumvent the immune defenses...Full Text Available
Mechanism-based inhibition of cytochrome P450 (CYP) 3A4 is characterized by NADPH-, time-, and concentration-dependent enzyme inactivation, occurring when some drugs are converted by CYPs to reactive...Full Text Available
Na+ channels are often spliced but little is known about the functional consequences of splicing. We have been studying the regulation of Na+ current inactivation in an...Full Text Available
During sexual development, Neurospora crassa inactivates genes in duplicated DNA segments by a hypermutation process, repeat-induced point mutation (RIP). RIP introduces C:G to T:A...Full Text Available
A search for homologs of the Bacillus subtilis PhoP response regulator in the group A streptococcus (GAS) genome revealed three good candidates. Inactivation of one of these, recently...Full Text Available
The safety of tissue allografts has come under increased scrutiny due to recent reports of allograft-associated bacterial and viral infections in tissue recipients. We report that 50 kGy of gamma irradiation, nearly three times the dose currently used, is an effective pathogen inactivation method when used under optimized conditions that minimize damage to the tissue. Cancellous bone dowels treated with a radioprotectant solution and 50 kGy of optimized irradiation had an ultimate compressive strength and modulus of elasticity equal to conventionally irradiated (18 kGy) and non-irradiated control bone grafts. We subjected bone dowels treated with this pathogen inactivation method to an in vitro cytotoxicity test using three different mammalian cell lines and concluded that the treated grafts were not cytotoxic. The log reduction of nine pathogens spiked into radioprotectant-treated bone irradiated to 50 kGy was also tested. We achieved 4.9 logs ...
Designer natriuretic peptides (NPs) represent an active area of drug development. In canine and human studies, the designer natriuretic peptide CD-NP demonstrated more desirable therapeutic potential than recombinant B-type NP (BNP), which is known as nesiritide and is approved for treatment of acute decompensated heart failure. However, why CD-NP is more effective than BNP is not known. We previously reported that CD-NP is a poorer activator of human guanylyl cyclase-A (GC-A) and a better activator of human guanylyl cyclase-B than BNP. Here, guanylyl cyclase bioassays were used to compare the susceptibility of CD-NP verses ANP, BNP, CNP and DNP to inactivation by human kidney membranes. The half time (t(1/2)) for CD-NP inactivation was increased by factors of 13, 3 and 4 compared to ANP, BNP and CNP, respectively, when measured in the same assay. Surprisingly, DNP failed to undergo complete inactivation and was the most ...
The issue of the duration of immunity, particularly for the modified live viral components of veterinary vaccines, has been a significant part of the recent vaccination debate. One manufacturer has increased the recommended booster interval for these components to 3 years give name and another now states `up to 4 years' immunity.There remain many unanswered questions regarding this duration of immunity (DOI). Studies suitable for data sheet claims are time consuming and costly and can only be performed in laboratory dogs under tightly controlled conditions. Evidence from rabies serology testing in the UK shows that the response of individual animals to routine vaccination is highly variable. Much of the published field evidence on the persistence of antibody titres originates from North Am...
In this paper we present an overview of the literature on efficacy and safety trials of the various pneumococcal conjugate vaccines on the market (PCV7) and in development (PCV9, PCV11 and allegedly PCV10 and PCV13), as well as of observations from post-licensure studies. Seven- (PCV7) and nine-valent PCV (PCV9) are reported to be sufficiently immunogenic after administration of a 3+1 schedule in infants in various RCTs. PncOMPC (PCV7 with a protein of N. meningitidis as a carrier) is less immunogenic, though this may have no repercussions for the protective efficacy against clinical disease. PCV7 is 82-97% efficacious against vaccine serotype (VT) IPD, 90% efficacious against (clinically diagnosed) pneumococcal pneumonia, and, like the 11-valent PCV, 57% efficacious against VT acute otiti...
Oral vaccination is the most desirable immunization method by which to prevent fish disease occurring at the early larval stage. We developed an Escherichia coli-based oral nervous necrosis virus (NNV) subunit vaccine and demonstrated its effectiveness in orange spotted grouper (Epinephelus coioides) fry. However, the incubation period was determined to be over 2weeks. To enhance the efficacy and shorten the incubation period, a safe and digestible biological adjuvant should be found. Vibrio anguillarum, a common marine bacterium with immune-stimulatory capability was selected as an expression host. An expression vector containing its heat shock protein 60 gene (groE) as an inducible promoter was constructed to express recombinant NNV coat protein as an antigen. The recombinant V. anguilla...
Recommended treatment of chronic hepatitis B with interferon-α and/or nucleos(t)ide analogues does not lead to a satisfactory result. Induction of HBV-specific T cells by therapeutic...Full Text Available
The trematode Fasciola hepatica (Fa.he.) is a common parasite of human and livestock. The hemoglobin (Hb) of Fa.he., a potential immunogen, was chosen for characterization...Full Text Available
BackgroundTrypanosoma cruzi, the etiologic agent of Chagas Disease, is a major vector borne health problem in Latin America and an emerging...Full Text Available
AbstractIndividual informed consent is a key ethical obligation for clinical studies, but empirical studies show that key requirements are often not met. Common recommendations to strengthen...Full Text Available
In the absence of a vaccine, there is an urgent need for the development of safe and effective topical microbicides to prevent the sexual transmission of human immunodeficiency virus type 1 (HIV-1)....Full Text Available
Genetically identical rhesus monkeys would have tremendous utility as models for the study of human disease and would be particularly valuable for vaccine trials and tissue transplantation studies where...Full Text Available
BackgroundPolychlorinated biphenyls (PCBs) may cause immunotoxic effects, but the detailed dose–response relationship and possible vulnerable time windows of exposure are...Full Text Available
A preventive medicine program of rubella control for trainees at the Air Force Military Training Center, Lackland AFB, Texas, was begun in October 1977. Incoming trainees were screened for rubella susceptibility,...Full Text Available
...broad range of experience in designing and building research and development laboratories for products such as vaccines, blood and sera, biological products, medical devices, solid dosage forms , highly potent products and cytotoxic material. We also have experience in a range of modular laboratories for TSE research, ...
A prospective cohort study was undertaken in two commercial California dairies. The treatment group, 246 cows, received three doses of a whole cell bacterin of J5 Escherichia coli (mutant of E. coli...Full Text Available
BackgroundSurrogate immunologic markers for natural and vaccine-mediated protection against tuberculosis (TB) have not been identified.MethodsHIV-infected...Full Text Available
In vivo plasminogen responses to various stimuli were studied. Plasminogen-125I was prepared and used first for metabolic studies of plasminogen in control dogs. The average results were:...Full Text Available
Cholera vaccine candidate Peru-15 was derived from a Vibrio cholerae O1 El Tor Inaba strain by deleting the cholera toxin genetic element, introducing the gene encoding cholera toxin B subunit into recA, and screening for nonmotility. In a controlled study, Peru-15 (2 x 10(8) cfu) was administered to 11 volunteers. No vaccinee developed diarrhea, and 10 of 11 had > 4-fold rises in vibriocidal antibody titers. One month later, 5 vaccinees and 5 control volunteers were challenged with wild type V. cholerae O1. Four of 5 controls developed diarrhea (mean, 1.9 L). Two Peru-15 vaccinees developed diarrhea, 1 with < 0.3 L and 1 with approximately 1.0 L; this latter volunteer had not developed a significant vibriocidal immune response to vaccination. Peru-15 shows promise as a single-dose, oral cholera vaccine that is safe, immunogenic, and protective. PMID:7561195
Severe acute respiratory syndrome (SARS) is an infectious disease caused by a novel coronavirus that cost nearly 800 lives. While there have been no recent outbreaks of the disease, the threat...Full Text Available
Antibodies against feline leukemia virus (FeLV) and the feline oncornavirus-associated cell membrane antigen (FOCMA) were transferred from pregnant cats to their suckling kittens. All of these kittens...Full Text Available
... were erroneous. The oriental rat flea, Xenopsylla cheopis Rothschild, is considered to be the primary vector of ... laboratory and production of Weigl's exanthematous typhus vaccine. In Maintenance of hum...
In this report we define the parameters of the human immune response after immunization with hepatitis B vaccine. 2 wk after booster immunization, there is significant spontaneous secretion of antibody...Full Text Available
The use of dendritic cells (DCs) as a cellular adjuvant is a promising approach to the immunotherapy of cancer. It has previously been demonstrated that DCs pulsed ex vivo with Toxoplasma gondii...Full Text Available
Increasing numbers of human cowpox virus infections that are being observed and that particularly affect young non-vaccinated persons have renewed interest in this zoonotic disease. Usually causing...Full Text Available
BackgroundThe objective of the study was to analyse the evolution of Bordetella pertussis population and the influence of herd immunity in different areas of the...Full Text Available
Evaluation of the protective efficacy of recombinant T-cell-reactive proteins of Coccidioides posadasii in a murine model of coccidioidomycosis has led to the discovery of potential...Full Text Available
Definition of the T-lymphocyte responses to dengue viruses should aid in the development of safe and effective vaccines and help to explain the pathophysiology of dengue hemorrhagic fever and dengue...Full Text Available
The identification of the protein targets for dengue virus-specific T lymphocytes may be useful for planning the development of subunit vaccines against dengue. We studied the recognition by murine...Full Text Available
vaccination card during trial enrollment at the KEMRI/CDC site in Kenya. (Alan Rubin, KEMRI) KEMRI Clinical Officer Paul Ogai reviews a prospective participant's...
Public health activities, especially infectious disease control, depend on effective teamwork. We present the results of a pilot audit questionnaire aimed at assessing the quality of public health services...Full Text Available
... A better understanding of the links between nutrients, the gut microbiome and the intestinal immune system is likely to pave the way to the development of innovative approaches that target activation of specific immunological pathways for promoting the hostrsquo;s gut defense. Such knowledge should positively impact on the development of vaccine formulations and intervention strategies for improved control of enteric infections in developing countries. Registration fees # The Registration Fee is $715.00 on and before September 7, 2011. # After September 7, 2011, the ...
Phosphoenolpyruvate carboxykinase (GTP) (PEPCK) specifically utilizes a guanosine or inosine nucleotide as a substrate, yet it does not share extended sequence homology with other GTP-binding proteins, and the molecular basis for its nucleotide specificity is not understood. In an effort to locate the enzyme's nucleotide-binding site, the authors have studied the interaction of cytosolic PEPCK from rat liver with the photoprobe 8-azidoGTP, which fulfills the criteria of a specific photoaffinity label for PEPCK. The photoprobe binds reversibly to the enzyme prior to modification and at low concentrations causes greater than 60% inactivation-GTP provides nearly complete protection against inactivation by 8-azidoGTP, whereas phosphoenolpyruvate and metal ions provide partial protection. In addition, the photoprobe is a substrate for the enzyme and has a K_m similar to that for GTP. However, the extent of covalent modification by ["3"2P]8-azido-GTP ...
Phosphoenolpyruvate carboxykinase (GTP) (PEPCK) specifically utilizes a guanosine or inosine nucleotide as a substrate, yet it does not share extended sequence homology with other GTP-binding proteins, and the molecular basis for its nucleotide specificity is not understood. In an effort to locate the enzyme's nucleotide-binding site, the authors have studied the interaction of cytosolic PEPCK from rat liver with the photoprobe 8-azidoGTP, which fulfills the criteria of a specific photoaffinity label for PEPCK. The photoprobe binds reversibly to the enzyme prior to modification and at low concentrations causes greater than 60% inactivation-GTP provides nearly complete protection against inactivation by 8-azidoGTP, whereas phosphoenolpyruvate and metal ions provide partial protection. In addition, the photoprobe is a substrate for the enzyme and has a K{sub m} similar to that for GTP. However, the extent of covalent modification by ...
Correlates between immunological parameters and protection against Bacillus anthracis infection in animals vaccinated with protective antigen (PA)-based vaccines could provide surrogate markers to evaluate the putative protective efficiency of immunization in humans. In previous studies we demonstrated that neutralizing antibody levels serve as correlates for protection in guinea pigs (S. Reuveny et al., Infect. Immun. 69:2888-2893, 2001; H. Marcus et al., Infect. Immun. 72:3471-3477, 2004). In this study we evaluated similar correlates for protection by active and passive immunization of New Zealand White rabbits. Full immunization and partial immunization were achieved by single and multiple injections of standard and diluted doses of a PA-based vaccine. Passive immunization was carried out by injection of immune sera from rabbits vaccinated with PA-based vaccine prior to ...
In this paper we present an overview of the literature on efficacy and safety trials of the various pneumococcal conjugate vaccines on the market (PCV7) and in development (PCV9, PCV11 and allegedly PCV10 and PCV13), as well as of observations from post-licensure studies. Seven- (PCV7) and nine-valent PCV (PCV9) are reported to be sufficiently immunogenic after administration of a 3+1 schedule in infants in various RCTs. PncOMPC (PCV7 with a protein of N. meningitidis as a carrier) is less immunogenic, though this may have no repercussions for the protective efficacy against clinical disease. PCV7 is 82-97% efficacious against vaccine serotype (VT) IPD, 90% efficacious against (clinically diagnosed) pneumococcal pneumonia, and, like the 11-valent PCV, 57% efficacious against VT acute otitis media. Naturally, it would be of paramount public health interest if the same levels of efficacy and effectiveness could be achieved with fewer doses. ...
Crude N,N-dialkylcarbamoylmethylphosphonates and phosphine oxide extractants, and particularly crude dihexyl N,N-diethylcarbamoylmethylphosphonate and octylphenyl N,N-diisobutylcarbamoyl phosphine oxide, are purified by distilling the crude materials in a thin film evaporator. Preferably, the crude materials are reacted with concentrated hydrochloric acid and then with aqueous sodium hydroxide prior to distillation in the thin film evaporator to hydrolyze impurities contained in them. The purified extractants are useful for extracting actinides and lanthanides from liquid waste streams. (author).
Crude N,N-dialkylcarbamoylmethylphosphonates and phosphine oxide extractants, and particularly crude dihexyl N,N-diethylcarbamoylmethylphosphonate and octylphenyl N,N-diisobutylcarbamoyl phosphine oxide, are purified by distilling the crude materials in a thin film evaporator. Preferably, the crude materials are reacted with concentrated hydrochloric acid and then with aqueous sodium hydroxide prior to distillation in the thin film evaporator to hydrolyze impurities contained in them. The purified extractants are useful for extracting actinides and lanthanides from liquid waste streams. (author).
Sewage gas or biogas as a product of anaerobic digestion contains methane and carbon dioxide as its main components. Biogas is scrubbed off carbon dioxide in absorption columns and purified to natural-gas quality. The purified gas then is piped to the gas pressure control station and fed into the public network. Description of process and plant is given.
The annual production of phosphate rock, on the order of about 30-40 million tons yearly, represents several million pounds of uranium. The present invention provides a process of purifying uranium tetrafluoride hydrate to produce a uranium (VI) peroxide product meeting 'yellow cake' standards using a double precipitation procedure. A fluoride complexing agent is used in the precipitation.
Wildland fire suppression personnel employed by the CDF do not currently have the equipment to protect themselves from the short-term acute affects of smoke from wildland fires. In addition, no regulations exist that specify appropriate respiratory protection and the current air-purifying respirator technology and carbon monoxide monitoring has not been adapted to fit wildland fire suppression requirements. This three-year limited study evaluated the ability of wildland fire fighters to perform their normal job function while wearing full-face air-purifying respirators. In the first two years of this study we designed, developed and field tested a prototype ``smart`` air-purifying respirator which incorporated a real-time carbon monoxide monitor into a commercial full-face respirator.` Data on carbon monoxide exposure while fighting wildland fires was collected. During the third year of this study we evaluated eight ...
The effects of eight chemicals on the inactivation rate of ultraviolet radiation on the colony building capabilities of two strains of Saccharomyces cervisae - a wild type strain and a mutant deficient in excision repair - were studied. The insecticide methoxychlor, the herbicide 2,4-dichlorophenoxyacetic acid, the fungicide pentachlorophenol and its metabolite tetrachlorohydroquinone, as well as the chemicals acrylonitrile and 2,3-dichloro-1-propene have no significant impact on the effects of UV radiation in Saccharomyces cerevisae. Depending on the concentration, trichloroethylene increases the sensitivity to UV radiation. The herbicide paraquat provides efficient protection against UV radiation at concentrations where a toxic effect cannot be observed even without UV. The results were rather similar for both strains. (orig.).
Relative doses of 75 MeV/u "1"2C ion beam after passing through different thickness of degraders were measured with the double ionization chamber method. The degrader's thickness equalized to penetration depth of the beam in water and its Bragg curve were obtained. The measured Bragg curve is compared with that calculated based on a model set up by the authors. Meanwhile, the inactivation effect of the ion beam passing through different thickness of degraders on melanoma (B_1_6) cells was measured, and the corresponding survival fraction data for B_1_6 cells were acquired
The 50th anniversary of Mary Lyon?s 1961 Nature paper, proposing random inactivation in early embryonic life of one of the two X chromosomes in the cells of mammalian females, provides an opportunity to remember and celebrate the work of those involved. While the hypothesis was initially put forward by Lyon based on findings in the mouse, it was founded on earlier studies, notably the work of Susumu Ohno; it was also suggested independently by Beutler and colleagues using experimental evidence from a human X-linked disorder, glucose-6-phosphate dehydrogenase deficiency, and has proved to be of as great importance for human and medical genetics as it has for general mammalian genetics. Alongside the hypothesis itself, previous cytological studies of mouse and human chromosomes, and the obse...
The removal phenol and 2-chlorophenol using soybean seed-hulls in the presence of hydrogen peroxide was demonstrated. The performance of a stirred membrane reactor containing soluble purified SEP was compared with a batch stirred reactor containing raw soybean seed-hulls. The purified enzyme reactor proved to be ineffective while good results were obtained with the crude seed-hulls for the removal of phenol and 2-chlorophenol. A single batch reactor containing raw seed-hulls was effective in removing more than 98.5% of 2-chlorophenol (initially at 10000 ppm) in less than 15 min. The performance of these rectors is comparable to existing HRP-based technology. The stability of the soybean peroxidase (SBP) enzyme was also examined in the presence of detergents (SDS, Tween 20 and Triton X-100).
Phycocyanin - a major phycobiliprotein constitutively produced by many cyanobacteria hold several promising applications in diagnostics, biomedical research, and therapeutics. This paper discusses a novel rapid method for the purification of cyanobacterial phycocyanin (C-PC) from Phormidium fragile using hydrophobic interaction chromatography. The protein was extracted and concentrated by grinding under liquid nitrogen and ammonium sulfate fractionation. C-PC was purified by single step hydrophobic interaction chromatography. Purified phycocyanin showed absorbance maximum (lmax) at 624nm. The criterion of purity (R) achieved was 4.52. Phycocyanin to phycoerythrin and phycocyanin to allophycocyanin purity ratio were 3.85 and 7.49, respectively. The purified protein showed a pI of 5.2 and ha...
Brief summaries are given of research carried out in the following areas: absorption spectroscopy of titanium, niobium, and molybdenum in uranium alloys; atomic absorption spectroscopy of selenium in stainless steel, computer program improvements for spectrophotometer control, and detection of beryllium in stack gases; electrolysis for inactivation of bacteria in cooling-tower water; improved solvent analysis by gas chromatography; sequential radionuclide separation for various sample matrices; progress on an electron microprobe analyzer; and miscellaneous projects.
The yeast phosphatidylinositol transfer protein (Sec14p) is required for biogenesis of Golgi-derived transport vesicles and cell viability, and this essential Sec14p requirement is abrogated by inactivation...Full Text Available
The rapidly activating and inactivating voltage-dependent outward K+ (Kv) current, IA, is widely expressed in central and peripheral neurons. IA has long been...Full Text Available
Nonthyroidal illness syndrome (NTIS) is a state of low serum 3,5,3′ triiodothyronine (T3) that occurs in chronically ill patients; the degree of reduction in T3 is associated...Full Text Available
Effects of gamma irradiation on hygienic quality and extraction yields in twenty-one kinds of Korean medicinal herbs were investigated. Gamma irradiation at 5-10 kGy inactivated contaminating microorganisms. The total extraction yield in fifteen kinds of the investigated medicinal herbs increased by 5-25% by a dose of 10 kGy. (author)
Cyclin-Dependent Kinase 1 (CDK1) is the major M-phase kinase known also as the M-phase Promoting Factor or MPF. Studies performed during the last decade have shown many details of how CDK1 is regulated...Full Text Available
The tumor suppressor p53 is often inactivated in breast cancer cells because the overexpression of its repressors (e.g., MDM2 and MDMX). Restoration of p53 activity by small molecules through counteracting...Full Text Available
Suicide substrate β, γ-bidentate Rh(III)ATP (RhATP) was used to map the metal ion-binding site in yeast phosphoglycerate kinase (PGK). Cleavage of the RhATP-inactivated enzyme with...Full Text Available
All Streptococcus pneumoniae strains isolated from paediatric clinical samples at Heraklion University General Hospital in the 10-year period 2000-2009 were tested for serotype and susceptibility to antimicrobials. Among a total of 258 strains, 159 were isolated in the 5-year period 2000-2004, before the introduction of the heptavalent pneumococcal conjugate vaccine (PCV7), and 99 in the post-PCV7 5-year period 2005-2009. The prevalence of PCV7-included serotypes decreased in the post-PCV7 period (p?=?0.0002), but an increase was observed for serotypes 7F (p?=?0.002) and 19A (p?=?0.004). Pan-susceptibility rates and susceptibility to cotrimoxazole increased in the post-PCV7 period (p?=?0.01 and p?=?0.008, respectively), but serotype 19A emerged as a contributor to multi-resistance (p?=?0.007). PCV7 was followed by decreased S. pneumoniae resistance and prevalence of vaccine-related serotypes but increased prevalence of serotypes 7F and 19A. ...
Based on studies on the genetic and molecular basis of Shigella flexneri invasive properties, we have constructed and evaluated a double mutant of S. flexneri serotype 5 for utilization as a live attenuated oral vaccine against shigellosis. The first mutation, icsA, blocks intracellular spread of bacteria as well as cell-to-cell infection. It affects the capacity of the invasive pathogen to form large abscesses in epithelia. The second mutation, iuc, eliminates production of the siderophore aerobactin thus impairing growth of the bacterium within tissues. This double mutant, SC5700 appeared safe when administered intragastrically to macaque monkeys as three doses (5 x 10(10) c.f.u. each) at weekly intervals. Protection against a challenge by the wild type isolate (M90T) was observed 4 weeks after the last vaccine inoculation. Duration of carriage was considerably reduced as compared to the control group in which all animals had developed severe ...
Vascular permeability factor (VPF) is an Mr 40-kD protein that has been purified from the conditioned medium of guinea pig line 10 tumor cells grown in vitro, and increases fluid permeability from blood...Full Text Available
Equine herpesvirus type 1 was cultivated in swine testis cell cultures and partially purified by differential centrifugation and centrifugation in a linear sucrose density gradient. The viral envelope...Full Text Available
N alpha-Acetylenkephalin carboxypeptidase was co-purified with N-acetyltyrosine deacetylase from monkey kidney. Almost 90% of the activity from the homogenate was recovered in a high-speed supernatant...Full Text Available
The LW blood group antigens reside on a 42-kDa erythrocyte membrane glycoprotein that was purified by immunoaffinity and partially sequenced. From this information, a specific PCR-amplified DNA fragment...Full Text Available
Several dissimilatory, sulfate-reducing bacteria were isolated from the rumen fluid of sheep fed purified diets containing sulfate. One isolate, strain D, was selected for characterization. This organism...Full Text Available
An immunoassay employing 125I-labeled enterotoxin B and polystyrene tubes coated with specific antibody was used for assaying purified and crude enterotoxin. Antibody was adsorbed to untreated...Full Text Available
Purified beta-toxin from Clostridium perfringens type C was found to be a single polypeptide chain protein with a molecular weight of approximately 30,000. The toxin was heat labile, with 75% of its...Full Text Available
Purified Shiga toxin (Stx) alone is capable of producing systemic complications, including hemolytic-uremic syndrome (HUS), in animal models of disease. Stx includes two major antigenic forms (Stx1...Full Text Available
The chloroplast and cytoplasmic isoenzymes of phosphoglycerate kinase (PGK) (EC. 2.7.2.3) from Hordeum vulgare leaves have been separated and purified for the first time to apparent...Full Text Available
The ruminal selenomonad strain H18 grew rapidly (mu = 0.50 h-1) in a defined medium containing glucose, ammonia, purified amino acids, and sodium (95 mM); little if any ammonia was utilized as a nitrogen...Full Text Available
mRNA guanylyltransferase has been extensively purified from calf thymus. A GTP-binding assay was used based on the observations by Shuman and Hurwitz (1981) and Venkatesan and Moss (1982) that vaccinia...Full Text Available
An enzyme active against carboxymethyl cellulose (CMC) was purified from the stationary-phase-culture supernatant of Clostridium josui grown in a medium containing ball-milled cellulose. The purification...Full Text Available
In a process for separating insoluble red mud from Bayer process streams the improvement is described which comprises contacting and mixing a Bayer process stream with a tertiary polyamine having a molecular weight of at least about 10,000 in an amount effective to reduce the iron content thereof.
BackgroundIt is important to generate biofuels and society must be weaned from its dependency on fossil fuels. In order to produce biofuels, lignocellulose is pretreated and the...Full Text Available
Polyamine oxidase was purified to homogeneity from leaves of water hyacinth by the criterion of sodium dodecyl sulfate gel electrophoresis (SDS disc PAGE). The enzyme showed a high specificity for spermidine...Full Text Available
Phosphoglycerate kinase (PGK) from the hyperthermophilic bacterium Thermotoga maritima has been purified to homogeneity. A second larger enzyme with PGK activity and identical N-terminal sequence was...Full Text Available
A cholinesterase was partially purified from bush bean (Phaseolus vulgaris L.) roots by using acridinium-based ligand affinity chromatography. The procedure gave a 78-fold increase in specific activity,...Full Text Available
Aminopeptidase II, one of the two major aminopeptidases in the giant alga Chara australis, was partially purified. Its molecular weight was estimated to be about 80,000 by gel permeation...Full Text Available
Cd-binding protein was extracted from tomato roots and purified on QAE-Sephadex A-25 and on Sephadex G-75 in 1 molar KCl buffer. The protein preparation was light brown and contained predominantly Cd...Full Text Available
A highly purified preparation of double-stranded RNA, obtained from virus-like particles in Penicillium cultures, was found to affert humoral immune responses in mice differentially depending on its...Full Text Available
All molybdoenzyme activities are absent in chlB mutants because of their inability to synthesize molybdopterin guanine dinucleotide, which together with molybdate constitutes the molybdenum cofactor...Full Text Available
Two bacteriolytic enzymes secreted by Achromobacter lyticus M497-1 were purified and identified as being very similar (considering their amino acid composition and N-terminal sequence) to alpha- and...Full Text Available
An antiserum was raised in a rabbit against highly purified human liver dihydropteridine reductase (EC 1.6.99.7). Dihydropteridine reductase from human liver, in human cultured fibroblasts and in continuous...Full Text Available
Antibodies of the IgM type present in rabbit anti-epiglycanin antiserum were purified by (NH4)2SO4 precipitation and by ion-exchange, affinity and gel-filtration chromatography. After papain treatment...Full Text Available
Pyrophosphate:fructose-6-phosphate phosphotransferase (PFP) was purified over 500-cold from endosperm of germinating castor bean (Ricinus commiunis L. var. Hale). The kinetic properties...Full Text Available
[Leu]- and [Met]enkephalin from thoracic ganglia of the shore crab Carcinus maenas have been purified to homogeneity by a reversed-phase HPLC procedure. Automated gas-phase sequencing revealed a primary...Full Text Available
Bisulfite reductase (desulfoviridin) and an assimilatory sulfite reductase have been purified from extracts of Desulfovibrio vulgaris. The bisulfite reductase has absorption maxima...Full Text Available
Although integration generally is considered a critical step in the retrovirus life cycle, it has been reported that visna virus, which causes degenerative neurologic disease in sheep, can productively...Full Text Available
The Moloney murine leukemia virus (MMLV) belongs to the Retroviridae family of enveloped viruses, which is known to acquire minute amounts of host cellular proteins both on the surface...Full Text Available
Phenylalanine hydroxylase was purified from crude extracts of human livers which show enzyme activity by usine two different methods: (a) affinity chromatography and (b) immunoprecipitation with an...Full Text Available
High-density lipoproteins (HDL) were examined as potential carriers of small peptides in plasma. HDL purified by density gradient centrifugation was delipidated and fractionated by size-exclusion...Full Text Available
Human Factor VIII/von Willebrand factor, purified by calcium citrate-cellulose chromatography and 4% agarose gel filtration was subjected to sodium dodecyl sulfate gel electrophoresis on gels containing...Full Text Available
NG2 chondroitin sulfate proteoglycan is a surface marker of oligodendroglial progenitor cells (OPCs) in various species. In contrast to well-studied rat OPCs, however, we found that purified...Full Text Available
AIMS: To evaluate the sensitivity of the Roche Cobas, Roche Amplicor plate kit, ligase chain reaction (LCR), and an in house polymerase chain reaction (PCR) by titration of purified elementary bodies...Full Text Available
Enolase is a multifunctional enzyme that plays important roles in many biological and disease processes. α-Enolase from human liver (hENO1) was expressed as a soluble protein and purified by...Full Text Available
Malate synthase (EC 4.1.3.2), an enzyme unique to the glyoxylate cycle, was purified to homogeneity from cotyledons of 72-hours, darkgrown cotton (Gossypium hirsutum L.) seedlings....Full Text Available
Lipopolysaccharides from different bacteria isolated from cotton were purified and chemically analyzed. Their pulmonary toxicity to animals was tested in inhalation tests. Lipopolysaccharides from Agrobacterium...Full Text Available
A pigment-dispersing hormone (PDH) from eyestalks of the fiddler crab Uca pugilator has been purified by gel filtration, ion-exchange chromatography, partition chromatography, and reversed-phase...Full Text Available
Recombinant mouse UDP-glucose pyrophosphatase (UGPPase), encoded by the Nudt14 gene, was produced in Escherichia coli and purified close to homogeneity. The...Full Text Available
Venomous mammals are rare, and their venoms have not been characterized. We have purified and characterized the blarina toxin (BLTX), a lethal mammalian venom with a tissue kallikrein-like activity...Full Text Available
An extracellular polysaccharide was purified from culture supernatants of Paenibacillus jamilae CP-7, a gram-positive bacillus that was isolated from compost prepared with olive mill...Full Text Available
The extracellular domain of human fibroblast growth factor receptor (XC-FGF-R) was expressed in Escherichia coli. The protein was purified to homogeneity and the interaction with basic fibroblast growth...Full Text Available
The chlorophyll fluorescence characteristics of mesophyll and bundle-sheath thylakoids from plant species with the C4 dicarboxylic acid pathway of photosynthesis were investigated using flow cytometry....Full Text Available
The acetylcholine receptor from the electric tissue of Torpedo californica is a large, integral membrane protein containing four different types of polypeptide chains. The structure of the purified...Full Text Available
Background and AimsPlants from the family Lemnaceae are widely used in ecological engineering projects to purify wastewater and eutrophic water bodies. However, the biology of nutrient...Full Text Available
1. ATP sulphurylase from Saccharomyces cerevisiae was purified 140-fold by using heat treatment, DEAE-cellulose chromatography and Sepharose 6B gel filtration. 2. The enzyme was stable...Full Text Available
From Toki-shakuyaku-san, an herbal formulation for “cleansing stagnated blood,” a key gene regulatory compound was purified and identified through a screening based on DNA microarray...Full Text Available
Purified cell walls were prepared from roots of Horse bean (Vicia faba L., var. minor) and Yellow Lupine (Lupinus luteus L.). Two methods were used:...Full Text Available
NAD(P)H:rubredoxin oxidoreductase (NROR) has been purified from the hyperthermophilic archaeon Pyrococcus furiosus. The enzyme is exceedingly active in catalyzing the NADPH-dependent...Full Text Available
An enantio-specific polyphenol oxidase (PPO) was purified ≈1,700-fold to apparent homogeneity from the creosote bush (Larrea tridentata), and its encoding gene was cloned. The...Full Text Available
A simple and rapid procedure was developed for the purification of cyclotron-produced {sup 203}Pb via the {sup 203}Tl(d,2n) {sup 203}Pb reaction. A Pb(II) selective ion-exchange resin, with commercial name Pb Resin from Eichrom Technologies, Inc., was used to purify {sup 203}Pb from the cyclotron-irradiated Tl target with excellent recovery of the enriched Tl target material. The purified {sup 203}Pb was used to radiolabel the monoclonal antibody Herceptin. The in vitro and in vivo properties of the {sup 203}Pb radioimmunoconjugate were evaluated.
This study was carried out mainly to produce uranium trioxide (UO_3), matching standard commercial specification from rock phosphate deposits in Uro and Kurun at eastern part of the Nuba Mountains. A simplified hydrometallurgical procedure has been adopted for production of yellow cake from the ore. The powdered ore sample was leached with concentrated H_2SO_4 acid with and without addition of KCIO_3 as an oxidant. The crude yellow cake was precipitated from the resulting green solution of phosphoric acid as Na_2U_2O_7 and (NH_4)_2U_2O_7 and subsequently purified by TBP extraction (tributylphosphate) and hydrogen peroxide as UO_4.2H_2O. TBP purified product was dried and calcined to UO_3 whereas UO_4.2H-2O was dried and reduced to UO_3 by Na_2S_2O_3. Prior to precipitation of crude yellow cake, Fe in the phosphoric acid solution was precipitated using magnesia. Elemental analysis has shown that the ore is rich in Ca and deficient in elements of ...
European sea bass (Dicentrarchus labrax L.) is a marine species of great economic importance, particularly in Mediterranean aquaculture. However, numerous pathogenic viruses, bacteria, fungi and parasites affect the species, causing various infectious diseases and thereby leading to the most heavy losses in aquaculture production of sea bass. In this respect, knowledge on molecular and genetic mechanisms of resistance to pathogens and specific features of immune response against various infectious agents should greatly benefit the development of effective vaccines and proper vaccination strategies in marker-assisted selection of fish resistant to a range of infections. To date, genetic knowledge on sea bass immune regulatory genes responsible for resistance to pathogens is relatively poor ...
Vaccine-mediated prevention of primary HIV-1 infection at the heterosexual mucosal portal of entry may be facilitated by highly optimised formulations or drug delivery devices for intravaginal (i.vag) immunization. Previously we described hydroxyethylcellulose (HEC)-based rheologically structured gel vehicles (RSVs) for vaginal immunization of an HIV-1 vaccine candidate, a soluble recombinant trimeric HIV-1 clade-C envelope glycoprotein designated CN54gp140. Here we investigated the efficacy of lyophilized solid dosage formulations (LSDFs) for prolonging antigen stability and as i.vag delivery modalities. LSDFs were designed and developed that upon i.vag administration they would reconstitute with the imbibing of vaginal fluid to mucoadhesive, site-retentive semi-solids. Mice were immunize...
AbstractBackground Randomised controlled trials (RCTs) in the 1970s and early 1980s showed the polysaccharide serogroup A vaccine (SgAV) prevented serogroup A meningococcal meningitis (SGAMM). Subsequent non-RCTs suggested significant variations in the age-specific duration of protection among children. Objectives To determine the protective effect, duration of protection, age-specific effects and the effect of booster doses in children of the SgAV against SGAMM. Search strategy We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2010, issue 2) which contains the Acute Respiratory Infections Group's Specialised Register, MEDLINE (January 1950 to May Week 3, 2010) and EMBASE (January 1974 to May 2010). Selection criteria We included RCTs. Non-RCTs ...
The plerocercoid stage of the tapeworm Spirometra mansonoides produces a functional analog of human growth hormone (hGH). Among the similarities between plerocercoid growth factor (PGF) and hGH is competition for the same receptors on rabbit liver membranes. To take advantage of this characteristic in a purification scheme for PGF, rabbit liver microsomes were solubilized in Triton X-100 and the hGH receptors were purified over an hGH affinity column. The purified receptors from six rabbit livers were coupled to Affi-Gel-10 to create a receptor affinity column which was used to purify PGF. Chromatography of crude PGF over the receptor column resulted in a 1044 fold increase in specific activity. SDS-PAGE in the presence of 2-mercaptoethanol showed that the affinity-purified PGF contained three protein bands with apparent Mrs of 27.5 K, 22 K, and 16.7 K. Injections of the ...
This study was carried out mainly to produce uranium trioxide (UO_3), with the standard of commercial specifications from rock phosphate deposits in eastern part of Nuba mountains, south Kurdufan state. A simplified hydrometallurgical procedure has been adopted for production of yellow cake from the ore. Elemental analysis has shown that the ore is rich in Ca and deficient in elements of potential interest such as Fe, Cu and Zn. Uranium content in ore, phosphoric acid and purified yellow cake (UO_3) obtained with different precipitants was analyzed using alpha-spectrometry. On the average, the activity concentration of uranium in ore corresponds to 82 #+-# 24 ppm (0.10%). From the data of pregnant liquor, it was observed that the addition of KCIO_3 as an oxidant improves the dissolution of uranium from the ore by almost 20%. Data has also indicated that the yellow cake purified by hydrogen peroxide has higher concentration of uranium by 44.5% ...
Serum and salivary responses of 95 infants to either a standard (4 x 10(4) plaque-forming units (PFU), 47 neonates) or a high dose (4 x 10(5) PFU, 48 neonates) of tetravalent reassortant rhesus rotavirus...Full Text Available
The ERA strain of rabies virus was propagated in a baby hamster kidney cell line (BHK-21/C13). The viral titer was 10(1.8) tissue culture infective doses (TCID) higher than that of commercial ERA vaccine....Full Text Available
To assess the influence of a live attenuated oral vaccine against porcine post-weaning colibacillosis (PWC) induced by enterotoxigenic Escherichia coli (ETEC) on mucosal lymphoid cell CD45 isoforms expression, experimental group of weaned pigs (n=6) was immunized orally with F4ac+ non-ETEC strain (day 0) and challenged with F4ac+ ETEC strain 7 days latter. Non-immunized ETEC-infected pigs (n=6) served as control. All pigs were killed on post-challenge day 7. The small intestine was excised for isolation of jejunal lamina propria (JLP) and ileal Peyer's patch (IPP) lymphocytes and immunohistochemical studies. The results obtained by immunophenotyping of isolated cells show that the proportion of CD45RA+ and CD45RC+ JLP, but not IPP, cells were higher in the non-ETEC-immunized ETEC-infected pigs versus non-immunized infected. Additionally, while CD45RA+ JLP cells increased only slightly, the expression of CD45RC isoform on the JLP cells was significantly higher ...
Two plasmids which express either nearly intact or truncated filamentous hemagglutinin (FHA) from Bordetella pertussis and which are marked with a tetracycline resistance (Tcr) gene were transformed into Salmonella dublin SL1438, an aroA deletion mutant intended for use as an attenuated oral vaccine against salmonellosis. These S. dublin recombinants, when fed to mice, induced serum immunoglobulin, immunoglobulin M (IgM), and sometimes IgA antibody responses to FHA and S. dublin. In addition, IgA antibodies against FHA were found in gut wash fluids. S. dublin carrying pDB2300, a multicopy plasmid encoding truncated FHA protein, induced a better antibody response than did S. dublin carrying pDB2000, a low-copy-number plasmid encoding full-sized FHA. Administration of tetracycline to mice enhanced the stability of recombinant plasmids, and tetracycline-treated mice developed higher anti-FHA titers. Although neither strain examined is suitable for use in a human oral ...
Newly hatched chicks were dosed orally with a Salmonella typhimurium wild-type strain, an S. enteritidis wild-type strain, and a genetically defined S. enteritidis aroA vaccine candidate, strain CVL30....Full Text Available
Type I interferon is important in anti-viral responses and in coordinating the innate immune response. Here we explore the use of interferon-β to adjuvant the response to a rabies virus...Full Text Available
BackgroundAfrican horse sickness virus (AHSV) causes a non-contagious, infectious disease in equids, with mortality rates that can exceed 90% in susceptible horse populations....Full Text Available
Among important candidates for babesial vaccines are apical complex proteins, including rhoptry-associated protein 1 (RAP-1) from Babesia bovis and B. bigemina, which...Full Text Available
A recent clinical trial of a T-cell-based AIDS vaccine delivered with recombinant adenovirus type 5 (rAd5) vectors showed no efficacy in lowering viral load and was associated with increased risk of...Full Text Available
In order to conserve vaccines in hot countries a solar refrigerator has been made by a French university. It is a photothermal refrigerator without batteries. It uses two gases: ammonia and calcium chloride. The refrigerator is autonomous and can work eleven days without sun. (N.C.)
Although the subtrate binding properties of adenylate kinase (AK) have been studied extensively by various biochemical and biophysical techniques, it remains controversial whether uncomplexed adenosine 5'-triphosphate (ATP) binds to the adenosine 5'-monophosphate (AMP) site of AK. The authors present two sets of experiments which argue against binding of ATP to the AMP site. (a) /sup 31/P nuclear magnetic resonance titration of ATP with AK indicated a 1:1 stoichiometry on the basis of changes in coupling constants and line widths. This ruled out binding of ATP to both sites. (b) ATP and MgATP were found to behave similarly by protecting AK from spontaneous inactivation while AMP showed only a small degree of protection. Such inactivation could also be protected or reversed by dithioerythritol and is most likely due to oxidation of sulfhydryl groups, one of which (cysteine-25) is located near the MgATP site. The results support ...
Microbial population and moisture content of traditional herbal medicines contaminated with 3 kinds of aerobic microbes, packed in 5 kinds of plastic packaging materials, followed by irradiation at minimum dose of 5 kGy and stored for 6 months were investigated. The highest reduction of microbial counts during storage was observed on samples packed in polyethylene bags. All of packaging materials used were found to be impermeable to microbes and water vapour. Radiation and packaging materials used acted synergistically to inactivate microbes durind storage. The microbial counts decreased as much as 2 to 4 log cycles during storage. (author).
Biosorption is an important removal mechanism in treating hydrophobic toxic organics in biological reactors. Equilibrium sorption isotherms and sorption kinetics of 2,4-dichlorophenol on live and chemically inactive anaerobic granules were studied. A metabolic inhibitor was used to inactivate the biological activity of the biomass. Results showed that the difference in the biosorption of live and chemically inactive anaerobic granules is not significant. This would suggest that anaerobic biosorption is mainly a physical-chemical process and that metabolic-mediated diffusion in the process is negligible. 12 refs., 3 tabs., 7 figs.
The grafted human glioblastoma cell CB109 was used as a model for intralesional therapy with 131I-labelled hyaluronectin glycoprotein (131I-HN). 131I-HN bound specifically to in situ hyaluronic acid (HA), a main component of the extracellular matrix which is involved in tumour invasion. Labelling experimental conditions were determined and, finally, 25 {mu}Ci/{mu}gHN, 1 {mu}g chloramine-T/{mu}gHN and a 60-s stirring period provided a 131I-HN preparation with an optimal affinity for HA (64% compared to unlabelled HN). Following intratumoral injection, 131I-HN was retained with a limited diffusion outside the tumour. On day 4 the radioactivity concentrated in the tumour was still 25 times greater than that in the liver, spleen and kidneys combined. For therapeutic assays, 65 {mu}Ci 131I-HN was injected into the tumour, resulting in a delivery of 6.8 Gy over a 7-day period. Controls received unlabelled HN, heat-inactivated HN, a mixture of ...
In situ assembly states of rat adipocyte glucose transport protein in plasma membrane (PM) and in microsomal pool (MM) were assessed by measuring target size (TS) of D glucose-sensitive, cytochalasin B binding activity. High energy radiation inactivated the binding in both PM and MM by reducing the total capacity of the binding (B/sub T/) without affecting the dissociation constant (K/sub D/). The reduction in B/sub T/ as a function of radiation dose was analyzed based on classical target theory, from which TS was calculated. TS in the PM of insulin-treated adipocytes was 58 KDa. TS in the MM of noninsulin-treated and insulin-treated adipocytes were 112 and 109 KDa, respectively. With MM, however, inactivation data showed anomalously low radiation sensitivities at low radiation doses showing a shoulder in the semilog plots, which may be due to an interaction with a radiation sensitive inhibitor. With these results, they propose the following ...
Objective:? Primary hyperparathyroidism (HPT) is characterised by autonomous secretion of PTH from enlarged parathyroid glands leading, in most patients, to asymptomatic hypercalcaemia. Familial hypocalciuric hypercalcaemia (FHH) is an autosomal dominant disorder caused by inactivating mutations in the calcium sensing receptor (CaSR) gene; it is characterised by lifelong and usually asymptomatic hypercalcaemia. Establishing the correct diagnosis is important because surgery can be curative in HPT, but ineffective in FHH. There is overlap in the diagnostic criteria for the two disorders and some patients carrying inactivating mutations in the CaSR gene, which is suggestive of FHH, also have HPT with hyperplastic parathyroid glands or adenomas. Design and Patients:? CaSR gene mutations were analyzed and clinical and biochemical parameters evaluated in 139 consecutive out-patients presenting with hypercalcaemia and suspected of having HPT. ...
The affinity label 8-((4-bromo-2,3-dioxobutyl)thio)adenosine 5{prime}-triphosphate (8-BDB-TA-5{prime}-TP) reacts covalently with rabbit muscle pyruvate kinase, incorporating 2 mol of reagent/mol of enzyme subunit upon complete inactivation. Protection against inactivation is provided by phosphoenolpyruvate, K{sup +}, and Mn{sup 2+} and only 1 mol of reagent/mol of subunit is incorporated. The authors have now identified the resultant modified residues. After reaction with 8-BDB-TA-5{prime}-TP at pH 7.0, modified enzyme was incubated with ({sup 3}H)NaBH{sub 4} to reduce the carbonyl groups of enzyme-bound 8-BDB-TA-5{prime}-TP and to introduce a radioactive tracer into the modified residues. Following carboxymethylation and digestion with trypsin, the radioactive peptides were separated on a phenylboronate agarose column followed by reverse-phase high-performance liquid chromatography in 0.1% trifluoroacetic acid with an acetonitrile gradient. ...
A multifunctional protein kinase, purified from rat liver as ATP-citrate lyase kinase, has been identified as a glycogen synthase kinase. This kinase catalyzed incorporation of up to 1.5 mol of and)2numberSPO4/mol of synthase subunit associated with a decrease in the glycogen synthase activity ratio from 0.85 to a value of 0.15. Approximately 65-70% of the TUPO4 was incorporated into site 3 and 30-35% into site 2 as determined by reverse phase high performance liquid chromatography. This multifunctional kinase was distinguished from glycogen synthase kinase-3 on the basis of nucleotide and protein substrate specificities. Since the phosphate contents in glycogen synthase of sites 3 and 2 are altered in diabetes and by insulin administration, the possible involvement of the multifunctional kinase was explored. Glycogen synthase purified from diabetic rabbits was phosphorylated in vitro by this multifunctional kinase at only 10% of the rate ...
The acute effects of injections of the human growth hormone-like factor purified from plerocercoids of the tapeworm Spirometra mansonoides on carbohydrate, lipid, and protein metabolisms were determined in intact rats. Male rats were injected ip with saline, insulin, or various doses of partially purified PGF. The rats injected with insulin had significantly reduced serum glucose concentrations but no dose of PGF caused a change in serum glucose levels. Insulin and PGF stimulated [14C]glucose and [14C]leucine oxidation to 14CO2 in adipose tissue and muscle and increased incorporation of both [14C]glucose carbons into lipids and [14C]leucine into protein in fat and muscle. The responses to PGF were dose-dependent and persisted after 3 hr of incubation in vitro. Injections of naloxone prior to injecting PGF to block the stress response did not prevent the stimulation of insulin-like responses by PGF. Therefore, PGF has intrinsic insulin-like ...
We describe a method for affinity purification of sequence-specific DNA binding proteins that is fast and effective. Complementary chemically synthesized oligodeoxynucleotides that contain a recognition site for a sequence-specific DNA binding protein are annealed and ligated to give oligomers. This DNA is then covalently coupled to Sepharose CL-2B with cyanogen bromide to yield the affinity resin. A partially purified protein fraction is combined with competitor DNA and subsequently passed through the DNA-Sepharose resin. The desired sequence-specific DNA binding protein is purified because it preferentially binds to the recognition sites in the affinity resin rather than to the nonspecific competitor DNA in solution. For example, a protein fraction that is enriched for transcription factor Sp1 can be further purified 500- to 1000-fold by two sequential affinity chromatography steps to give Sp1 of an estimated 90% ...
A simple and rapid procedure was developed for the purification of cyclotron-produced "2"0"3Pb via the "2"0"3Tl(d,2n) "2"0"3Pb reaction. A Pb(II) selective ion-exchange resin, with commercial name Pb Resin from Eichrom Technologies, Inc., was used to purify "2"0"3Pb from the cyclotron-irradiated Tl target with excellent recovery of the enriched Tl target material. The purified "2"0"3Pb was used to radiolabel the monoclonal antibody Herceptin. The in vitro and in vivo properties of the "2"0"3Pb radioimmunoconjugate were evaluated.
We have developed a process for chemical purification of carbon nanotubes for solution-processable thin-film transistors (TFTs) having high mobility. Films of the purified carbon nanotubes fabricated by simple drop coating showed carrier mobilities as high as 164 cm2V?1s?1, normalized transconductances of 0.78 Sm?1, and on/off current ratios of 106. Such high performance requires the preparation of a suspension of micrometer-long and highly purified semiconducting single-walled carbon nanotubes (SWCNTs). Our purification process includes length and electronic-type selective trapping of SWCNTs using recycling gel filtration with a mixture of surfactants. The results provide an important milestone toward printed high-speed and large-area electronics with roll-to-roll and ink-jet device fabri...
Cyclization of deproteinized natural rubber (DPNR) or purified natural rubber latex was effectively performed in latex phase by using trimethylsilyl-trifluoromethane sulfonate or trimethylsilyl triflate (TMSOTF) as a novel catalyst, which is still not reported in the case of natural rubber latex. Various cyclization conditions affecting the degree of cyclization were studied, such as dry rubber contents, temperature, TMSOTF concentrations, and time. The cyclized products were characterized by FTIR, Raman, 1H-, and 13C-NMR spectroscopies, as well as DSC and TGA. The degree of cyclization was estimated by 1H-NMR spectrum. It was found that the degree of cyclization in NR was a function of cyclization conditions. The thermal stability of cyclized DPNR increased with the degree of cyclization....
Summary Background The incidence of food allergy to wheat is increasing. Its diagnosis depends on the purity of major allergens and their inclusion in tests. Isolation and characterization of wheat allergens are therefore of utmost importance. Objective To purify and identify wheat flour allergens most frequently recognized by patients' IgE antibodies and to study their allergenicity. Methods Water/salt-soluble extracts from wheat flour were prepared and separated using a combination of ultrafiltration, isoelectric focusing and liquid chromatography. Purified proteins were analysed by immunoblotting using pooled sera from patients with atopic dermatitis who possessed IgE specific to wheat. Wheat proteins found to bind IgE were subsequently identified by matrix-assisted laser desorption/ion...
Containing an epidemic at its origin is the most desirable mitigation. Epidemics have often originated in rural areas, with rural communities among the first affected. Disease dynamics in rural regions have received limited attention, and results of general studies cannot be directly applied since population densities and human mobility factors are very different in rural regions from those in cities. We create a network model of a rural community in Kansas, USA, by collecting data on the contact patterns and computing rates of contact among a sampled population. We model the impact of different mitigation strategies detecting closely connected groups of people and frequently visited locations. Within those groups and locations, we compare the effectiveness of random and targeted vaccinations using a Susceptible-Exposed-Infected-Recovered compartmental model on the contact network. Our simulations show that the targeted vaccinations of only 10% ...
Marburg virus belongs to the genus Marburgvirus in the family Filoviridae and causes a severe hemorrhagic fever, known as Marburg hemorrhagic fever (MHF), in both humans and nonhuman primates. Similar to the more widely known Ebola hemorrhagic fever, MHF is characterized by systemic viral replication, immunosuppression and abnormal inflammatory responses. These pathological features of the disease contribute to a number of systemic dysfunctions including hemorrhages, edema, coagulation abnormalities and, ultimately, multiorgan failure and shock, often resulting in death. A detailed understanding of the pathological processes that lead to this devastating disease remains elusive, a fact that contributes to the lack of licensed vaccines or effective therapeutics. This article will review the...
The heavy water reactor such as Wolsung No.1 and No.2 has a purification system to purify the reactor coolant. The control system regulates the coolant temperature to protect the ion exchanger. After the fuel exchanges of operating plant, the increase of the coolant pressure makes the purification temperature control difficult. In this paper, the controllability of the control dynamics of the purification system was analysed and the optimal parameters were proposed. To reduce the effects of the flow disturbance, the feedforward control structure was proposed and analysed.
The modes of action of the five major endo-(1----4)-beta-D-glucanases (I, II, III, IV and V) purified from Penicillium pinophilum cellulase were compared by h.p.l.c. analysis, with normal, 1-3H-labelled...Full Text Available
We have studied the self-association reactions of purified GDP-liganded tubulin into double rings and taxoid-induced microtubules, employing synchrotron time-resolved x-ray solution scattering. The...Full Text Available
NADP:malic enzyme from corn (Zea mays L.) leaves was purified by conventional techniques to apparent homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis....Full Text Available
In this study, Leydig cells were purified from 70 day-old Sprague Dawley male rats and incubated with 10 and 100 μg/mL of methanol extract of Basella alba (MEBa) for 4 hours...Full Text Available
Bovine paratuberculosis is caused by infection of young calves with Mycobacterium avium subsp. paratuberculosis. In some of the chronically infected cows the long asymptomatic...Full Text Available
The major core protein (p28) of MMC-1, an endogenous type C virus of the rhesus monkey (Macaca mulatta), was purified and subjected to structural and immunological analyses. The NH2-terminal amino acid...Full Text Available
Besarhanamides A (1) and B (2) are fatty acid amides purified from the marine cyanobacterium, Lyngbya majuscula, collected from Pulau Hantu, Singapore. The structure determination of these secondary metabolites was carried out using extensive 2D NMR spectral data as well as chemical manipulations including the Marfey's method. In addition, besarhanamide A exhibited moderate toxicity with LD50 at 13mM in the brine shrimp toxicity bioassay.
The mission of the Heavy Water portion of D Area (or 400 Area) at SRS is to purify the site inventory of heavy water for storage in the Reactor Areas for future DOE missions.
Platelet-rich plasma (PRP) is a platelet concentrate that has widely been used to accelerate the healing of soft and hard tissues. The preparation of PRP has been described by several authors. Preparation protocols vary from system to system, depending on the concentration of different integrating proteins. The objective of this article is to describe the principal use of PRP mixed with fat grafts in aesthetic plastic surgery.
We have purified and characterized two peptides, named KAaH1 and KAaH2 (AaH polypeptides 1 and 2 active on K+ channels, where AaH stands for Androctonus australis Hector),...Full Text Available
Abstract The Hippo pathway is an evolutionally conserved protein kinase cascade involved in regulating organ size in vivo and cell contact inhibition in vitro by governing cell proliferation and apoptosis. Deregulation of the Hippo pathway is linked to cancer development. Its first core kinase Warts was identified in Drosophila more than 15 years ago, but it gained much attention when other core components of the pathway were identified 8 years later. Major discoveries of the pathway were made during past several years. The core kinase components Hippo, Salvador, Warts, and Mats in the fly and Mst1/2, WW45, Lats1/2, and Mob1 in mammals phosphorylate and inactivate downstream transcriptional co-activators Yorkie in the fly, Yes-associated protein (YAP) and transcriptional co-activator with ...
Heart cells from the clam Ruditapes decussatus were routinely cultured with a high level of reproducibility in sea water based medium. Three cell types attached to the plastic after 2?days and could be maintained in vitro for at least 1?month: epithelial-like cells, round cells and fibroblastic cells. Fibroblastic cells were identified as functional cardiomyocytes due to their spontaneous beating, their ultrastructural characteristics and their reactivity with antibodies against sarcomeric ?-actinin, sarcomeric tropomyosin, myosin and troponin T-C. Patch clamp measurements allowed the identification of ionic currents characteristic of cardiomyocytes: a delayed potassium current (I K?slow) strongly suppressed (95%) by tetraethylammonium (1?mM), a fast inactivating potassium current (I K?fas...
AbstractBackground In recent years nano-metaloxides which easily penetrate into the cells with special interest due to their higher chemical reactivity as compared to that of similar materials in the bulk form. Of particular interest are nano-TiO2 and ZnO, which have been widely used for their bactericidal and anticancerous properties. Purpose The aim of the present study was to examine the bactericidal properties of nano-TiO2 and ZnO combined with visible light on S. aureus and S. epidermitis, known for their high prevalence in infected wounds. Study Using the technique of electron-spin resonance (ESR) coupled with spin trapping, we examined the ability of TiO2 and ZnO nanoparticle suspensions in water to produce reactive oxygen species (ROS) with and without visible light irradiation. Th...
Date palm (Phoenix dactylifera) is widely cultivated in Kutch district of Gujarat and the fruits are harvested at immature stage before the onset of monsoon to prevent spoilage. The immature date fruits with less commercial value were used for processing into date juice concentrate. Immature dates were crushed and treated with 0.1% pectinase enzyme for 120 min to obtain maximum juice. Date juice was found to be rich in reducing sugars (16.1%) and total sugars (18.3%). Juice was pasteurized at 85?C to inactivate the enzyme, cooled and centrifuged at 3000 rpm to get clear juice. The juice was concentrated in a thin film evaporator to a total soluble solids (TSS) of 76?Brix in 2 passes. Chemical composition of date juice during different stages of concentration was determined. Date juice conc...
Please cite this paper as: Ectopic mineralization of connective tissue in Abcc6-/- mice: effects of dietary modifications and a phosphate binder - a preliminary study. Experimental Dermatology 2008; 17: 203-207. Abstract: Pseudoxanthoma elasticum (PXE), a heritable multisystem disorder, is caused by mutations in the ABCC6 gene. We have developed a murine model for PXE by targeted inactivation of the corresponding mouse gene. A feature of this mouse model is ectopic mineralization of connective tissue capsule surrounding the bulb of vibrissae. This study was designed to investigate the effect of dietary sevelamer hydrochloride (Renagel), a phosphate binder, and specific mineral modifications on ectopic mineralization of connective tissue in Abcc6-/- mice. Three groups were fed a specific di...
Abstract Hereditary primary hyperparathyroidism (HPT) may develop as a solitary endocrinopathy (FIHP) or as part of multiple endocrine neoplasia Type 1, multiple endocrine neoplasia Type 2A, or hereditary HPT-jaw tumor syndrome. Inactivating germline mutations of the tumor suppressor gene CDC73 account for 14 and 50% of all FIHP and HPT-JT patients, respectively, and have also been found in almost 20% of apparently sporadic parathyroid carcinoma patients. Although more than 60 independent germline mutations have been described, to date no rearrangement affecting the CDC73 locus has been identified. By means of multiplex-PCR we found a large germline deletion affecting the whole gene in a two-generation HPT-JT family. Subsequently array-CGH and specific PCR analysis determined that the muta...
The moment analysis of pulse responses was applied to a biological activated carbon (BAC) bed to evaluate the microbial activity. Glucose and activated carbon fiber (ACF) were employed in experiments as a model tracer and an activated carbon respectively. No significant difference in biodegradation rates of glucose by microorganisms attached on different solid supports such as activated carbon inactivated carbon and glass fibers was observed. However, higher microbial activity was obtained from microorganisms supported on the ACF in terms of the apparent degradation of total organic including metabolic byproducts by microorganisms, since part of the metabolic byproducts was irreversibly deposited onto the ACF. 11 refs., 6 figs.
A fundamental study of the application of bacteria to the recovery of toxic heavy metals from aqueous environments was carried out. The biosorption characteristics of cadmium and lead ions were determined with purple nonsulfur bacteria, Rhodobacter sphaeroides and hydrogen bacteria, Alcaligenes eutrophus H16 that were inactivated by steam sterilization. A simplified version of the metal binding model proposed by Plette et al. was used for the description of meal binding data. The results showed that the biosorption of bivalent metal ions to whole cell bodies of the bacteria was due to monodentate binding to two different types of acidic sites: carboxilic and phosphatic-type sites. The number of metal binding sites of A. eutrophus was 2.4-fold larger than that of R. sphaeroides.
Abstract:- Campylobacter jejuni-is the leading cause of bacterial diarrheal illness worldwide. Many strains are now becoming multidrug resistant. Apple-based edible films containing carvacrol and cinnamaldehyde were evaluated for bactericidal activity against antibiotic resistant and susceptible-C. jejuni-strains on chicken. Retail chicken breast samples inoculated with D28a and H2a (resistant strains) and A24a (a sensitive strain) were wrapped in apple films containing cinnamaldehyde or carvacrol at 0.5%, 1.5%, and 3% concentrations, and then incubated at 4 or 23 C for 72 h. Immediately after wrapping and at 72 h, samples were plated for enumeration of viable-C. jejuni. The antimicrobial films exhibited dose- and temperature-dependent bactericidal activity against all strains. Films with ...
Nitroaromatic radiosensitizers become metabolically bound preferentially to hypoxic cells and at least 10/sup 9/ adducts/cell can be tolerated as non-toxic. EMT-6 tumor cells have been incubated in hypoxia in the presence of /sup 3/H-Misonidazole and /sup 125/I-Azomycin Riboside for various times and the amount of /sup 3/H or /sup 125/I bound/cell was determined. Cells were stored as monolayers at 25"0C for up to 96 hr to accumulate radioactive decays and transferred at various times to 37"0C for colony-forming assays. No radiation inactivation was measured in cells which had incorporated at least 10/sup 6/ /sup 3/H or 10/sup 5/ /sup 125/I atoms. Previous studies had shown that -- 1% of MISO adducts to EMT-6 cells was associated with cellular DNA. These data indicate that the radiation-induced damage produced by these quantities of bound /sup 3/H or /sup 125/I causes little or not cell inactivation. The results of current studies to measure the ...
Cdc7 kinase plays an essential role in firing of replication origins by phosphorylating components of the replication complexes. Cdc7 kinase has also been implicated in S phase checkpoint signaling downstream of the ATR and Chk1 kinases. Inactivation of Cdc7 in yeast results in arrest of cell growth with 1C DNA content after completion of the ongoing DNA replication. In contrast, conditional inactivation of Cdc7 in undifferentiated mouse embryonic stem (ES) cells leads to growth arrest with rapid cessation of DNA synthesis, suggesting requirement of Cdc7 functions for continuation of ongoing DNA synthesis. Furthermore, loss of Cdc7 function induces recombinational repair (nuclear Rad51 foci) and G2/M checkpoint responses (inhibition of Cdc2 kinase). Eventually, p53 becomes highly activated and the cells undergo massive p53-dependent apoptosis. Thus, defective origin activation in mammalian cells can generate DNA replication checkpoint signals. ...
The receptor occupancy-biological effect relationship for muscarinic receptors in guinea pig ileal smooth muscle has been studied by comparison of radioligand binding and contractile response. Muscarinic receptors in homogenates of ileal smooth muscle were labeled with (/sub 3/H)-1-Quinuclidinyl benzilate. Treatment with propylbenzilylcholine mustard (PrBCM), to inactivate irreversibly muscarinic receptors, caused a large dose dependent rightward shift of the dose-response curve to three agonistic furtrethonium derivatives with a concomitant decrease in maximal response. Using those data, the fraction of receptors remaining unoccupied (q-values) and true affinity constants (-log K/sub A/-values) were calculated. Exposure to 20 or 60 nM PrBCM for 15 minutes resulted in a 39% and a 61% reduction in specific (/sup 3/H)-1-Quinuclidinyl benzilate binding sites respectively to be compared with a 62% and a 85% decrease expected from calculated q-values. K/sub A/-values ...
Studies by the University of Lowell Radiation Laboratory and the US National Marine Fisheries Service N.E. Laboratory in Gloucester, MA on softshelled clams (Mya arenaria) demonstrated the effectiveness of low to medium doses of Cobalt 60 source gamma irradiation in the inactivation of Escherichia coli, Salmonella typhimurium, Staphylococcus aureus and Streptococcus faecalis. Post-irradiation survival and organoleptic studies when extended to hardshelled clams (Mercinaria mercenaria) and American oysters (Crassostrea virginica) showed no significant decline in consumer qualities or 6 day post irradiation survival in oysters at doses of up to 3.0 kGy. The capacities of the American oyster to sustain relatively high doses of gamma irradiation were demonstrated by 6 day post-exposure survivorship values of greater than 90% for samples receiving 3.0, 5.0 and 7.0 kGy. Initial studies of inactivation of Polio I virus and a simian rotavirus (SA-11) ...
We recently found that many RNA polymerase II transcription factors are modified with N-acetylglucosamine residues. These sugar moieties confer upon transcription factors an ability to bind the lectin wheat germ agglutinin. We have taken advantage of this interaction to devise a purification procedure for the "GC-box" binding transcription factor Sp1. Crude nuclear extracts are first subjected to wheat germ agglutinin affinity chromatography and then subjected to sequence-specific DNA affinity chromatography. The Sp1 protein purified by this procedure is at least 95% pure, and the overall recovery is greater than 80%. In addition to yielding larger quantities of Sp1 than conventional schemes, the new purification procedure is also simpler and more rapid. We show that wheat germ agglutinin affinity chromatography can also be used to purify the glycosylated forms of the CCAAT-binding transcription factor. Thus, wheat germ agglutinin affinity ...
A liquid column was used for the experiments reported in Part 1. However, it only gives the observation of the effect of fast reaction because the liquid flow was controlled to uniform laminar flow and the contact is limited to short time of around 10 ms. In practical absorbing operation, turbulence is involved in liquid flow, and the residence time for contact is long. Hence, the absorption of trace iodine in the purified air has been experimented by using a constant interface area type stirred absorption tank. Prior to the experiment, the characteristics of the absorption tank was investigated by conducting pure carbon dioxide absorption test with purified water. It gave the conclusion that the tank was sufficiently usable for fundamental researches. In short contact time absorption, the iodine dissolved and absorbed in liquid phase is affected by reaction of hypoiodous acid and poly-iodide ion formation due to hydrolysis by basic catalyst, ...
Comparisons were made between various LH and FSH radioimmunoassays currently being used to measure avian hormones. The two LH assays were the homologous chicken system of Follett et al. (1972) and the turkey assay of Burke et al. (1979). These assays were also used in heterologous arrangement by interchanging the iodinated LH fractions and antisera. Five FSH assays were analyzed: two homologous chicken systems (Scanes et al., 1977; Sakai and Ishii, 1980) an assay based on mammalian materials (rat FSH and anti-ovine FSH antiserum, and one using labelled turkey FSH (Burke et al., 1979) with an anti-chicken FSH antiserum. The potencies of purified chicken and turkey gonadotrophin preparations and of a range of plasma samples from Japanese quail were measured in each assay. The two LH systems showed some degree of species specificity, such that chicken LH was more active than turkey LH in the chicken assay, whereas the reverse was true in the turkey LH assay. The ...
Summary In this study, antimicrobial activity of zein films incorporated with partially purified lysozyme and disodium ethylenediaminetetraacetic acid (Na2EDTA) has been tested on selected pathogenic bacteria and refrigerated ground beef patties. The developed films containing 700-g-cm-2 lysozyme and 300-g-cm-2 Na2EDTA showed antimicrobial activity on Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella typhimurium. The application of lysozyme and Na2EDTA incorporated zein films on beef patties significantly decreased total viable counts (TVC) and total coliform counts after 5-days of storage compared to those of control patties (P-2EDTA or Na2EDTA alone significantly slowed down the oxidative changes in patties during storage (P-P-2EDTA for active packaging of refrigerated mea...
Purified protein kinase C (C-kinase) from guinea pig pancreas and rat brain stimulated adenylate cyclase activity in purified rat adipocyte membranes. Cyclase stimulation occurred over 100 to 1000 mU/ml of C-kinase activity, required greater than 10 ..mu..M calcium, proceeded without a lag, was not readily reversible, and required no exogenous phospholipid. Moreover, C-kinase inhibitors, such as chlorpromazine and palmitoyl carnitine, inhibited selectively adenylate cyclase which was activated by C-kinase and calcium. Depending on assay conditions, 10 nM 12-0-tetradecanoylphorbol-13-acetate (TPA) either enhanced or was required for kinase action on cyclase. Also, TPA plus calcium promoted the quantitative association of C-kinase with membranes. Adenylate cyclase activation by C-kinase was seen both in the presence and absence of exogenous GTP, indicating that the kinase effect does not result from an action on the GTP-binding, inhibitory ...
We describe a compact ultrafiltration system for the removal of pyrogens and bacteria from water labeled with the stable isotopes of deuterium and oxygen-18. The ultrafiltration system is constructed from readily available commercial components and can achieve complete removal of pyrogens and bacteria from 1L contaminated water within 30 min. By use of our procedure, loss of the isotopically labeled water by retention in the filtration system was minimal. The purified water is suitable for both oral and intravenous administration to healthy human subjects participating in nutrition studies.
Carrier-free "8"8Y radioisotope, which has the longest half life (T=106.6 d) of yttrium radioisotopes and is a #gamma#-ray emitter, was obtained by proton irradiation of strontium, followed by cooling for one month. Then, "8"8Y was purified by precipitating strontium as strontium nitrate and extracting yttrium with tri-n-butyl phosphate (TBP). The decontamination factor of strontium to yttrium was more than 4x10"3 and chemically pure yttrium radioisotope was obtained. (author) 6 refs.
We present a simple protocol to purify a coherent-state superposition that has undergone a linear lossy channel. The scheme constitutes only a single beam splitter and a homodyne detector, and thus is experimentally feasible. In practice, a superposition of coherent states is transformed into a classical mixture of coherent states by linear loss, which is usually the dominant decoherence mechanism in optical systems. We also address the possibility of producing a larger amplitude superposition state from decohered states, and show that in most cases the decoherence of the states are amplified along with the amplitude.
Methods and apparatus are described for efficient photodetachment and purification of negative ion beams. A method of purifying an ion beam includes: inputting the ion beam into a gas-filled multipole ion guide, the ion beam including a plurality of ions; increasing a laser-ion interaction time by collisional cooling the plurality of ions using the gas-filled multipole ion guide, the plurality of ions including at least one contaminant; and suppressing the at least one contaminant by selectively removing the at least one contaminant from the ion beam by electron photodetaching at least a portion of the at least one contaminant using a laser beam.
Low-dust flue gases can be washed efficiently of SO/sub 2/, and the flue gas heat can be utilized. The residual heat fraction still required by the process is characterized by a temperature of 15/sup 0/K above the temperature of the purified flue gas. Marketable crystalline ammonium sulfate is produced. The high efficiency and low heat consumption are achieved by combining all process stages in a single, fully integrated cycle process free of heat and materials losses. The process is characterized by high efficiency, low investment and operating costs, recovery of thermal energy, and production of an ammonium sulfate fertilizer.
Flow cytogenetics is a rapidly developing technology which complements rather than supplants the traditional methods of cytogenetic analysis. As an adjunctive approach to chromosome classification, flow systems measurements of isolated chromosomes give new information relative to the enrichment of A-T or G-C base sequences on specific chromosomes. As a result, chromosomal fluorescence polymorphisms both within and among individuals that are not always associated with banding polymorphisms can be detected. Finally, flow sorting provides bulk quantities of highly purified chromosomes for use in biochemical studies. Methodology is detailed. (PSB)
FuelCell Energy in Connecticut has sold 4.5 MW worth of power plants to California renewable energy company BioFuels Fuel Cells, part of New Energy Capital and BioFuels Energy. Three molten carbonate fuel cell power plants - a 2.8 MW DFC3000 unit, a 1.4 MW DFC1500 system, and a 300 kW DFC300 unit - will be installed at three locations in the San Diego area, and utilize purified biogas from the Point Loma wastewater treatment plant as their primary fuel source.
Biogas supply to the public gas grid is a new option discussed in the most recent publication of Fachagentur Nachwachsende Rohstoffe (FNR) e.V. The biogas is purified, upgraded to natural gas quality and fed into an existing gas grid. Once there, it can be transported across long distances and also used for electric power generation at the consumer side. The study investigates inhowfar and in what instances this method is feasible and promising. It discusses the technical background and the regional potential of biogas. Seven model biogas systems are presented as examples. (orig.)
Borrelia burgdorferi sensu lato genospecies were differentiated by PCR-based sequencing of the borrelial flagellin gene. To evaluate the usefulness of KingFisher"T"M magnetic particle processor in PCR product purification, borrelia PCR products were purified with KingFisher"T"M magnetic particle processor prior to cycle sequencing and the quality of the sequence data received was analyzed. KingFisher was found to offer a rapid and reliable alternative for borrelial PCR product purification.
An anaerobic fermenter-decanter for the purification of residual water from the sugar industry, with recovery of methane, consists of a tank with inclined walls, with a central agitator on a vertical shaft. A flexible cover anchored by its periphery to the walls of the tank and totally submerged forms a collecting pocket for the fermentation gases. The water to be purified is introduced, after being heated to about 35, towards the bottom of the tank near the agitator. A metal collecting bell with submerged edges and with the shaft of the agitator passing axially through it is connected by its edges to a central opening of the cover. The purification yields may exceed 90%.
Al, Ga and In complexing with methylthymol blue (H_6R) purified by gel filtration is studied. in the case of metal excess complexes with the ratio of components M:H_6R=2:1 with #lambda#=587(Al), 590(Ga) and 593 nm(In) appear. In the case of reacting agent excess complexes with the ratio of component 1:1 with #lambda#_m_a_x=480-490 (Al) and 480 nm(Ga, In) appear. The mechanism of complexing reactions is studied. Molar extinction coefficients and stability constants are calculated.
The author presents a new technique for production of yellow cake with double precipitation, thus solving a series of problems for precipitating uranium with traditional double precipitation. The new technique can not only remove ferric ions and sulfate radicals but also make solid-liquid separation easy, utilize effectively the sulfuric acid produced in ferric ions precipitation process, and increase uranium concentration of leaching liquor. To take it as producing yellow cake will save investment, simplify operation, and cut down the consumption of raw materials and energies. It is more competitive than ion-exchange or solvent extraction in the process of extracting-purifying and preparing yellow cake.
This issue has the following articles: (1) Deterrence with a Minimum Nuclear Stockpile - Commentary by Bruce T. Goodwin; (2) Enhancing Confidence in the Nation's Nuclear Stockpile - Livermore experts are participating in a national effort aimed at predicting how nuclear weapon materials and systems will likely change over time; (3) Narrowing Uncertainties - For climate modeling and many other fields, understanding uncertainty, or margin of error, is critical; (4) Insight into a Deadly Disease - Laboratory experiments reveal the pathogenesis of tularemia in host cells, bringing scientists closer to developing a vaccine for this debilitating disease. (5) Return to Rongelap - On the Rongelap Atoll, Livermore scientists are working to minimize radiological exposure for natives now living on or wishing to return to the islands.
Selected applications of novel techniques in Agricultural Biotechnology, Health Food formulations and Medical Biotechnology are being reviewed with the aim of unraveling future developments and policy changes that are likely to open new markets for Biotechnology and prevent the shrinking or closing of existing ones. Amongst the selected novel techniques with applications in both Agricultural and Medical Biotechnology are: immobilized bacterial cells and enzymes, microencapsulation and liposome production, genetic manipulation of microorganisms, development of novel vaccines from plants, epigenomics of mammalian cells and organisms, and biocomputational tools for molecular modeling related to disease and Bioinformatics. Both fundamental and applied aspects of the emerging new techniques are being discussed in relation to their anticipated, marked impact on future markets and present policy changes that are needed for success in either Agricultural or Medical ...
Abstract Development of inhibitors and vaccines that mitigate rumen-derived methane by targeting methanogens relies on knowledge of the methanogens present. We investigated the composition of archaeal communities in the rumens of farmed sheep (Ovis aries), cattle (Bos taurus) and red deer (Cervus elaphus) using denaturing gradient gel electrophoresis (DGGE) to generate fingerprints of archaeal 16S rRNA genes. The total archaeal communities were relatively constant across species and diets, and were less variable and less diverse than bacterial communities. There were diet- and ruminant-species-based differences in archaeal community structure, but the same dominant archaea were present in all rumens. These were members of three coherent clades: species related to Methanobrevibacter ruminan...
The efficiency of dendrosome (a gene porter) was assessed in transferring recombinant human rotavirus VP2 cDNA into A549, a human lung cell line. After gene transferring, transmission electron microscopy showed core-like particles (CLPs) formation in the transfected cells both with dendrosome and lipofectamine porters. In addition, western blotting analysis showed that the expression of VP2 gene was almost equal in the dendrosome and lipofectamine-transfected cells. Also, the cytotoxicity studies revealed that dendrosome had a lower cytotoxicity than lipofectamine. Therefore, our study may introduce dendrosome as a possible carrier for gene transferring into the human lung cell line, especially, for intranasally administration of DNA vaccines.
Mycoplasma ovipneumoniae is associated with chronic non-progressive pneumonia of sheep and goats. As with many other mycoplasmas involved in animal diseases, protective immune responses have not been achieved with vaccines, even though antibody responses can be obtained. This study focuses on characterizing the interaction of M. ovipneumoniae with ovine PBMC using carboxy-fluorescein-succinimidyl-ester (CFSE) loading and flow cytometry to measure lymphoid cell division. M. ovipneumoniae induced a strong in vitro polyclonal suppression of CD4^+, CD8^+, and B blood lymphocyte subsets. The suppressive activity could be destroyed by heating to 60^oC, and partially impaired by formalin and binary ethyleneimine treatment that abolished its viability. The activity resided on the surface-exposed m...
Three in vivo assays were used to measure the immunocompetence of chickens in two generations of a selection experiment. The obtained data were used to estimate the variance components for sire and dam for antibody production to Pasteurella multocida and Mycoplasma gallisepticum vaccines, for T-cell-mediated immunity evaluated by a phytohemagglutinin wing web assay, and for clearance of foreign particles from the circulatory system. Heritabilities of and genetic correlations among these immunological traits were calculated from the sire variance components. Heritability estimates of the immunological traits based on the sire component of variance ranged from .06 to .53, and genetic correlations among immunological traits were generally negative. PMID:1956847
Chitosan (a?-(1-4)-amino-2-deoxy-b?-D-glucan) is a deacetylated form of chitin, a polysaccharide from crustacean shells. Its unique characteristics, such as positive charge, biodegradability, biocompatibility, nontoxicity, and rigid structure, make this macromolecule ideal for an oral vaccine delivery system. We prepared reverse-phase evaporation vesicles (REVs) sandwiched by chitosan (Chi) and polyvinylic alcohol (PVA). However, in this method, there are still some problems to be circumvented related to protein stabilization. During the inverted micelle phase of protein nanoencapsulation, hydrophobic interfaces are expanded, leading to interfacial adsorption, followed by protein unfolding and aggregation. Here, spectroscopic and immunological techniques were used to ascertain the effects ...
Brucella melitensis is a Gram-negative coccobacillus bacteria belonging to the Alphaproteobacteria subclass. It is an important zoonotic pathogen that causes brucellosis, a disease affecting sheep, cattle, and sometimes humans. The B. melitensis strain M5-90, a live attenuated vaccine cultured from the B. melitensis virulent strain M28, has been an effective tool to control brucellosis in goats and sheep in China. Here we report the complete genome sequences of B. melitensis M28 and M5-90, strains with different virulence backgrounds, which will serve as a valuable reference for future studies. PMID:21478357
Background: Chloroquine has in-vitro activity against influenza and could be an ideal candidate for worldwide prevention of influenza in the period between onset of a pandemic with a virulent influenza strain and the development and widespread dissemination of an effective vaccine. We aimed to assess the efficacy of such an intervention. Methods: In this randomised, double-blind, placebo-controlled trial done at a single centre in Singapore, we randomly assigned (1:1) healthy adults to receive chloroquine phosphate (500 mg/day for 1 week, then once a week to complete 12 weeks) or matching placebo by use of a computer-generated randomisation list. Participants filled an online symptom diary every week, supplemented by daily diaries and self-administered nasal swabs when unwell. Haemagglutin...
Summary The antigenic relationship between the phocine distemper virus (PDV) strain causing the epidemic in 2002 and the PDV strain of 1988, canine distemper virus from two dogs and one marten, and one measles virus strain was investigated in vivo and in vitro using monospecific polyclonal and monoclonal antibodies directed against five different proteins of canine or phocine distemper virus (N, P, M, F, H). Epitopic mapping revealed no difference between the PDV strains causing the epidemics in 1988 or 2002. However, the use of these antibodies allowed discrimination between different morbilliviruses including a vaccine strain of canine distemper virus. The major differences among the investigated morbilliviruses were found in the H protein.
Using the phasmid vector pSL5, the genomic DNA fragment of T. aquaticus YT1 which contained the thermostable DNA polymerase (Taq-polymerase) gene was cloned. The BglII fragment of this genome locus was subcloned in the BamHI site of the pUC19 plasmid. To optimize the Taq-polymerase gene expression in E. coli cells, the gene was cloned in the correct reading frame regarding the initiation ATG codon of the pPR-TGATG-1 expression vector. The gene expression in this vector was controlled by the phage lambda PR promoter and the temperature-sensitive phage lambda repressor. We used PCR to amplify the short 5'-end fragment of the Taq-polymerase gene coding for the part into which an artificial SacI site was introduced. This site has been used for cloning the PCR product into the pPR-TGATG-1 vector, and the missing gene part was cloned into the KpnI site of the PCR product from the natural cloned gene. The cells of the E. coli PVG-A1 strain, which was obtained in the end, expressed efficiently ...
The thermostable Thermus aquaticus DNA polymerase (Taq Pol) has been the key factor in transforming the initial PCR method into one with huge impact in molecular biology and biotechnology. Therefore, the development of effective affinity adsorbents for the purification of Taq Pol, as well as other DNA polymerases, attracts the attention of the enzyme manufacturers and the research laboratories. In this report we describe a simple protocol for the purification of Taq Pol from E. coli lysates, leading to enzymes of high specific activity and purity. The protocol is based on a single affinity chromatography step, featuring an immobilized ligand selected from a structure-biased combinatorial library of dNTP-mimetic synthetic ligands. The ligand library was screened for its ability to bind and purify Taq Pol from E. coli lysates. One immobilized ligand (mABSGu) of the general formula X-Trz-Y, bearing 9-aminoethylguanine (AEGu) and aniline-2-sulfonic acid (mABS) linked ...
Photoaffinity labeling analogs of the adenylate cyclase activator forskolin (PF) have been synthesized, purified and tested for their effect on preparations of membrane-bound, Lubrol solubilized and forskolin affinity-purified adenylate cyclase (AC). All analogs of forskolin significantly activated AC. However, in the presence of 0.1 to 0.3 microM forskolin, the less active forskolin photoaffinity probes at 100 microM caused inhibition. This inhibition was dose-dependent for PF, suggesting that PF may complete with F for the same binding site(s). After cross-linking (125I)PF-M to either membrane or Lubrol-solubilized AC preparations by photolysis, a radiolabeled 100-110 kDa protein band was observed after autoradiography following SDS-PAGE. F at 100 microM blocked the photoradiolabeling of this protein. Radioiodination of forskolin-affinity purified AC showed several protein bands on autoradiogram, however, only one band ...
ZnMT has been postulated to function in essential metal homeostasis and metal detoxication. This study was undertaken to evaluate the potential of liver ZnMT to mediate the interaction of Zn and Pb with the Zn metalloenzyme, ALAD. Pretreatment of rats with Zn activated liver ALAD and attenuated the inhibition of ALAD by Pb in vitro and in vivo. Liver cytosol from Zn-pretreated rats injected with /sup 203/Pb was fractionated via gel filtration and anion-exchange chromatography and disclosed that both Zn and /sup 203/Pb co-eluted in the MT fractions. Both purified ZnMT-I and ZnMT-II bound /sup 203/Pb in vitro as shown by gel filtration chromatography. Addition of purified liver ZnMT to purified bovine liver ALAD reaction mixtures increased enzyme activity by 2-fold and prevented inhibition of ALAD by Pb. Addition of apothionein alone decreased the activity of Zn-activated ALAD and also attenuated Pb inhibition of the enzyme. ...
The Drosophila sequence-specific DNA binding protein, Adf-1, is capable of activating transcription of the alcohol dehydrogenase gene, Adh, and is implicated in the transcriptional control of other developmentally regulated genes. We have cloned the cDNA encoding Adf-1 by generating specific DNA probes deduced from partial amino acid sequence of the protein. Several cDNA clones encoding an extended open reading frame were isolated from a phage lambda library. The complete amino acid sequence of Adf-1 deduced from the longest cDNA reveals structural similarities to the putative helix-turn-helix DNA binding motif of Myb and Myb-related proteins. DNA sequence analysis of genomic clones and Northern blot analysis of mRNA suggest that Adf-1 is a single-copy gene encoding a 1.9-kb transcript. Purified recombinant Adf-1 expressed in Escherichia coli binds specifically to Adf-1 recognition sites and activates transcription of a synthetic Adh promoter in vitro in a manner ...
Abstract:- This study was undertaken to investigate the antimicrobial effect of organic acids against-Escherichia coli-O157:H7,-Salmonella-Typhimurium, and-Listeria monocytogenes-on whole red organic apples and lettuce. Several studies have been conducted to evaluate organic acids as sanitizers. However, no studies have compared antimicrobial effects of various organic acids on organic fresh produce, including evaluation of color changes of produce. Apples and lettuce were inoculated with a cocktail of 3 strains each of 3 foodborne pathogens provided above and treated with 1% and 2% organic acids (propionic, acetic, lactic, malic, and citric acid) for 0, 0.5, 1, 5, and 10 min. With increasing treatment time and acid concentration, organic acid treatments showed significant reduction compar...
Ionizing radiation, mainly in the form of #gamma#-rays from the radioisotope cobalt 60, is being used increasingly for the inactivation of contaminants in cosmetic and toiletry preparations. The treatment is applied to the product in its final pack, and therefore, it is important to recognize that the properties of the packaging materials could be affected, particularly if high radiation doses are used. The stability of a wide range of materials is discussed. The nature and extent of chemical and physical changes in plastics very much depends on the presence of antioxidants and other additives, and on the environmental conditions during irradiation. Some useful information is available from research into the use of radiation for the preservation of food - many films and laminates have been cleared for use in the United States. The low radiation doses used for the control of contaminants in cosmetic products are quite unlikely to cause changes in packaging materials ...
The iron hormone hepcidin is inhibited by matriptase-2, a liver serine-protease encoded by TMPRSS6 gene. Cleaving the BMP-coreceptor hemojuvelin, matriptase-2 impairs the BMP/SMAD signaling pathway, downregulates hepcidin and facilitates iron absorption. TMPRSS6 inactivation causes iron-deficiency-anemia refractory to iron administration both in humans and mice. Genome wide association studies have shown that the SNP rs855791, which causes the matriptase-2 V736A amino acid substitution, is associated with variations of serum iron, transferrin saturation, hemoglobin and erythrocyte traits. Here we show that in vitro matriptase-2 736(A) inhibits hepcidin more efficiently than 736(V). Moreover, in a genotyped population, after exclusion of samples with iron deficiency and inflammation, hepcidin, hepcidin/transferrin saturation and hepcidin/ferritin ratios were significantly lower and iron parameters were consistently higher in homozygotes 736(A) than in 736(V). Our ...
Irradiation of live bivalve molluscs is a promising new technique to reduce the public health threats of viral and bacterial pathogens in the consumption of raw shellfish. In studies conducted within the past year, live hardshelled clams and oysters were innoculated with Simian Rotavirus SA-11, Poliovirus I, and Hepatitis-A virus and treated with gamma ray ionizing radiation from University of Lowell/DOE 800,000 Curie "6"0Co source over a dose range of 0.5--10 kiloGray (50--1000 kilorad) to determine the effectiveness of the process in eliminating viral activity and thus easing the potential viral health threats associated with uncooked bivalves. Two viral strains, SA-11 and Poliovirus I, were employed as models to perfect laboratory technique and to estimate the most appropriate range of dose exposures necessary for optimal inactivation of the infectious pathogen, Hepatitis A. Post-irradiation survivorship of the oysters and quahogs was assessed over a range of ...
Conditioned medium from Sertoli cells, prepared from testes of 20-day-old rats, contains component(s) that inhibit the incorporation of (3H)-thymidine into DNA of peritubular myoid cells (PMC) and inhibit the proliferation of PMC. These components are trypsin-resistant, heat-stable compounds having a molecular weight less than 30,000. The active inhibitory components in Sertoli cell conditioned medium are inactivated by treatment with heparinase, but not by treatment with hyaluronidase or chondroitin sulfate lyases. Addition of heparin or heparan sulfate results in inhibition of DNA synthesis by PMC in a dose-dependent manner, whereas other glycosaminoglycans (GAGs) examined (hyaluronic acid, keratan sulfate, and chondroitin sulfate) have no detectable effects. Heparin and heparan sulfate are unique among GAGs tested in inhibiting the characteristic multilayer growth pattern of PMC following the attainment of confluence in serum-rich medium. On the basis of these ...
The phenomenon of the limit of pitting corrosion in direction to positive potentials is studied by potentiokinetic polarization after a jump in the transpassive range and by potentiostatic tests at technical wrought materials and at model alloys of the systems NiCrMo and NiMo in CaCl{sub 2} solution in the concentration range 1 to 9 mol/l chloride at pH-values of 1 to 9 at temperatures of 30 to 110 C. Surface-analytical investigations gives in connection with knowledges from anodic polarization studies directions to the mechanism of the limit of pitting corrosion. Ranges of the limit of pitting corrosion are obtained at materials with a Mo content above 6.5% and contents of chloride of the media above 2 mol/l chloride. Increasing temperatures, increasing contents of chloride and sulfate shift the potential of the limit of pitting corrosion being always above 0.2 V (SCE) at potentiostatic determination to noble direction. There are indications that the mechanisms of limit of pitting ...
Abstract Polycystic kidney diseases (PKD) are inherited disorders characterized by fluid-filled cysts primarily in the kidneys. We previously reported differences between the expression of Cux1, p21, and p27 in the cpk and Pkd1 null mouse models of PKD. Embryonic lethality of Pkd1 null mice limits its study to early stages of kidney development. Therefore, we examined mice with a collecting duct specific deletion in the Pkd1 gene. Cux1 was ectopically expressed in the cyst lining epithelial cells of newborn, P7 and P15 Pkd1CD mice. Cux1 expression correlated with cell proliferation in early stages of cystogenesis, however, as the disease progressed, fewer cyst lining cells showed increased cell proliferation. Rather, Cux1 expression in late stage cystogenesis was associated with increased ...
A tk{sup +/-} mouse embryonic stem (ES) cell line, designated 1G2, has been created in which one allele of the thymidine kinase (tk) gene was inactivated by targeted homologous recombination. This line is an analog of the mouse lymphoma tk{sup +/-} L5178Y cell line, which is used widely to assess the mutagenicity of chemical agents. Treatment of 1G2 cells with the alkylating agent N-ethyl-N-nitrosourea (ENU) resulted in a dose-related increase in tribluorothymidine-resistant colonies. Mutant frequencies of 152 and 296 per 10{sup 6} cells were determined for 0.1 and 0.3 mg/ml doses of ENU, compared with a spontaneous mutant frequency of 15 per 10{sup 6} cells. The data indicate that tk{sup +/-} 1G2 ES cells may be useful for the creation of a transgenic mouse model for assessing in vivo mutation using an endogenous autosomal gene. 45 refs., 2 figs., 1 tab.
Marketing of minimally processed vegetables (MPV) are gaining impetus due to its convenience, freshness and apparent health effect. However, minimal processing does not reduce pathogenic microorganisms to safe levels. Food irradiation is used to extend the shelf life and to inactivate food-borne pathogens. In combination with minimal processing it could improve safety and quality of MPV. A microbiological screening method based on the use of direct epifluorescent filter technique (DEFT) and aerobic plate count (APC) has been established for the detection of irradiated foodstuffs. The aim of this study was to evaluate the applicability of this technique in detecting MPV irradiation. Samples from retail markets were irradiated with 0.5 and 1.0 kGy using a {sup 60}Co facility. In general, with a dose increment, DEFT counts remained similar independent of the irradiation while APC counts decreased gradually. The difference of the two counts gradually increased with ...
Sphere-forming abilities in culture condition are considered a hallmark of cancer stem-like cells, which represents tumor cell invasiveness and stem-like characteristics. We aimed to show that the sphere-forming subpopulation of human malignant melanoma cell line WM-266-4 acts differently to zebrafish embryo extracts compared with their bulk counterpart. Spheres were maintained in neural stem cell culture conditions. The embryos of zebrafish at specific developmental stages were collected and the extracts were purified under 100 kDa. Spheres were treated with embyo extracts and proliferation assay and immunocytochemistry were conducted. Spheroid cells expressed nestin and epidermal growth factor receptor (EGFR) but not melanoma antigen recognized by T-cells (MART)1, indicating their stem-l...
Purification parameters of the vacuum distillation process for AM50A and AZ31B magnesium alloy were studied. The vacuum distillation tests were kept at 580 C to 620 C up to 10 h under a vacuum pressure of about 1 Pa. The purification ratio for AM50A magnesium alloy increases with increasing the purification time. The chemical compositions of materials deposited purifying at 580 C for AM50A magnesium alloy almost satisfy the values for pure magnesium specified by JIS standard specification. The chemical compositions of deposited materials at 600 C for AZ31B magnesium alloy satisfy the values specified by the standard specification except for Zn. (orig.)
The presence of two major aminopeptidases (aminopeptidases I and II) in the giant alga Chara australis was shown using polyacrylamide gel electrophoresis. Partially purified aminopeptidase I had a molecular weight of about 120,000, hydrolyzed both leucine-?-naphthylamide (pH optimum 6.0) and alanine-?-naphthylamide (pH optimum 7.5), and was located both inside and outside the vacuole. Aminopeptidase I was inhibited by p-chloromercuribenzoic acid, iodoacetic acid, 1,10-phenanthroline, and N-tosyl-l-phenylalanine chloromethyl ketone. Aminopeptidase II hydrolyzed alanine-?-naphthylamide but not leucine-?-naphthylamide and was located only outside the vacuole.ImagesFig. 1Fig. 2Fig. 4Fig. 5
Biological control agents can be more effective if their populations are genetically diverse, particularly when the target invasive plant comprises a range of genotypes with different susceptibilities and occurs across various microclimates. We report on the use of an efficient approach to find, in the native range, diverse isolates of a rust fungus for biological control. An outdoor trap garden containing various clones of invasive European blackberry (Rubus fruticosus agg.) collected in Australia, each with a different DNA phenotype, was established in France. Within 4?weeks of establishment, the leaf-rust fungus Phragmidium violaceum was recovered from all clones in the garden. Molecular analyses of eight recovered and purified isolates of the fungus from the garden revealed that they w...
The operation of the Bullhouse Minewater System, used to treat iron-rich minewaters discharged from long-abandoned coal mines, was studied to minimise pollution of the Upper Don river in the UK. Tests were carried out to determine optimal operating conditions. Ochre flocculation with polyacrylamide was efficient in batch tests as the ochre self-flocculated and sedimented in the lagoon without the need for chemical enhancement. The rate of accretion of the pipework in the system was modified by partial dewatering, reshaping and hardening of the deposits during ageing. Samples of ochre recovered from the drying beds and sump deposits were converted to ferric sulphate coagulant by dissolution with sulphuric acid and used with a polyelectrolyte to successfully purify reservoir water and pretreat wastewater. There was considerable local interest in the scheme with educational opportunities for schools and colleges.
ObjectiveExtended periods of muscle disuse, physical inactivity, immobilization, and bedrest result in a loss of muscle mass and a decrease in muscle force, which are accompanied by an increase in oxidative stress. We investigated the effects of the intake of green tea catechins on unloading-induced muscle dysfunction in tail-suspended mice. MethodsTen-week-old male BALB/c mice were fed a purified control diet or a diet containing 0.5% tea catechins for 14 d. Thereafter, the mice were subjected to continuous tail suspension for 10 d. On the final day, muscle mass, contractile force production, antioxidant potential, and carbonylated protein levels were evaluated. ResultsHind limb unloading caused a loss of soleus muscle weight and muscle force. Intake of tea catechins significantly inhibit...
Membranes in Palladium and its alloys, in particular Pd/Ag alloys, are selectively permeable to hydrogen and can therefore be used to purify hydrogen. It is intended to use them as material for electrodes in fuel cells. As the manufacturing process of a substrate Pd/Ag-layer consists of several stages and is still not completely understood, it would be advantageous to be able to characterise separately the layers of noble metals obtained by electro-plating . Attempts are being made in this work to vary the cristallographical structure of the deposited alloy by the choice of test parameters and by sintering when depositing the Pd/Ag. The layers produced are to be examined using an interference microscope and X-ray diffraction. Moreover, the diffusion behaviour of the hydrogen is to be examined with an electro-chemical pulse method specifically developed for this purpose. figs., tabs., 27 refs.
Summary Isomalto-oligosaccharides (IMO) belong to a group of prebiotics that can significantly increase the number of protective gut microflora. A one-step method using neopullulanase (NPN) in conjunction with saccharifying -amylase (SAA) for the bioconversion of rice starch into IMO was investigated. Purified rice starch slurry (30% w/w) was mixed with NPN (3.5 U g-1 starch substrate) and SAA (6.5 U g-1 starch substrate) and the slurry was incubated at 57 C for 92-h under constant stirring. The carbohydrate composition of the resulting syrup was analysed by high performance liquid chromatography (HPLC) and the dextrose equivalent (DE) determined by titration. The amount of IMO in the syrup reached maximum (59.2%, dry basis) after 72-h of bioconversion. The concentration of glucose and mal...
Mimivirus is the prototype of a new family (the Mimiviridae) of nucleocytoplasmic large DNA viruses (NCLDVs), which already include the Poxviridae, Iridoviridae, Phycodnaviridae and Asfarviridae. Mimivirus specifically replicates in cells from the genus Acanthamoeba. Proteomic analysis of purified mimivirus particles revealed the presence of many subunits of the DNA-directed RNA polymerase II complex. A fully functional pre-transcriptional complex appears to be loaded in the virions, allowing mimivirus to initiate transcription within the host cytoplasm immediately upon infection independently of the host nuclear apparatus. To fully understand this process, a systematic study of mimivirus proteins that are predicted (by bioinformatics) or suspected (by proteomic analysis) to be involved in...
We investigated the presence of XMRV in a cohort of Quebec patients with chronic fatigue syndrome (CFS). DNA was purified from activated peripheral blood mononuclear cells (PBMCs) and PCR was used to detect XMRV gag and env in 72 patients. Anti-XMRV antibodies were searched in sera of 62 patients by Western blot analysis. Attempts to detect XMRV antigens was made, using immunofluorescence with Gag anti-p30 antiserum on activated PBMC from 50 patients. Plasma viremia was measured by RT-PCR on 9 subjects. Finally, detection of infectious virus in 113 CFS subjects was made by co-culture of PHA+IL-2 activated PBMC with human LNCaP carcinoma cells, and by infecting the same susceptible cells with plasma, using a reverse transcriptase (RT) assay as a readout in both experiments. No detection of ...
Starch from the fruits of sweetsop (Anonna squamosa) and soursop (Anonna muricata) were isolated and purified and the fat, ash, phosphorus and protein contents measured. The amount of amylose present was determined spectrophotometrically and found to be very similar (19%) for both starches. Scanning electron microscopy showed very small indented and spherical granules from both with an average granule size of 4.84mm and 4.72mm, respectively. The physicochemical properties, namely the swelling power, solubility, pasting characteristics, paste clarity and freeze-thaw stability were studied to assess the functionality of the starch pastes as hydrocolloids. The sweetsop starch showed higher swelling power and solubility compared to soursop starch and had a lower gelatinization temperature indi...
Current studies in plants suggest that the content of the coenzyme NAD is variable and potentially important in determining cell fate. In cases that implicate NAD consumption, re-synthesis must occur to maintain dinucleotide pools. Despite information on the pathways involved in NAD synthesis in plants, the existence of a mitochondrial nicotinamide mononucleotide adenylyltransferase (NMNAT) activity which catalyses NAD synthesis from nicotinamide mononucleotide (NMN) and ATP has not been reported. To verify the latter assumed pathway, experiments with purified and bioenergetically active mitochondria prepared from tubers of Jerusalem artichoke (Helianthus tuberosus L.) were performed. To determine whether NAD biosynthesis might occur, NMN was added to Jerusalem artichoke mitochondria (JAM)...
The known biological functions of RNA have expanded in recent years and now include gene regulation, maintenance of sub-cellular structure, and catalysis, in addition to propagation of genetic information. As for proteins, RNA function is tightly correlated with structure. Unlike proteins, structural information for larger, biologically functional RNAs is relatively limited. NMR signal degeneracy, relaxation problems, and a paucity of long-range {sup 1}H-{sup 1}H dipolar contacts have limited the utility of traditional NMR approaches. Selective isotope labeling, including nucleotide-specific and segmental labeling strategies, may provide the best opportunities for obtaining structural information by NMR. Here we review methods that have been developed for preparing and purifying isotopically labeled RNAs, as well as NMR strategies that have been employed for signal assignment and structure determination.
SummaryBackgroundCandida albicans is one of the most important etiologic agents causing superficial and deep fungal infections. For prevention of candidiasis, it is important to develop a rapid system that discriminates C. albicans at the strain level.ObjectiveTo develop a system that can identify C. albicans at the strain level.MethodsGenomic DNAs were purified from 179 clinical isolates of C. albicans, and were used as templates for PCR amplification of 25S rDNA and ALT repeats in repetitive sequences (RPSs). PCR products generated from ALT repeats were digested with EcoRI and/or ClaI in order to study the relationships between restriction profiles and amplification profiles.ResultsOne hundred and seventy nine clinical isolates were grouped into genotypes A (92 isolates), B (38 isolates)...
The endophytic fungus Trichothecium roseum LZ93 from Maytenus hookeri was found to antagonize other pathogenic fungi in vitro. To identify which compound contributed substantially to the antagonism, we fermented the strain and purified its fermentation products. Eleven compounds were obtained, including two trichothecenes, five rosenonolactones, two cardiotonic cyclodepsipeptides, and two sterols. Compound 11?-hydroxyrosenonolactone (1) was assigned according to 1D and 2D-NMR data for the first time. At the same time, the 1H and 13C-NMR assignments for 6?-hydroxyrosenonolactone (2) were revised. Of all of them, only trichothecin (6) showed strong antifungal activity. Based on our observations of the antagonistic activity and the other experimental results, we suggest that the antifungal co...
Thioredoxin reductase (TrxR) isoforms play important roles in cell physiology, protecting cells against oxidative processes. In addition to its endogenous substrates (Trx isoforms), hepatic TrxR can reduce organic selenium compounds such as ebselen and diphenyl diselenide to their selenol intermediates, which can be involved in their hepatoprotective properties. Taking this into account, the aim of the present study was to evaluate the hypothesis that ebselen, diphenyl diselenide and its analogs (4,4'-bistrifluoromethyldiphenyl diselenide, 4,4'-bismethoxydiphenyl diselenide, 4.4'-biscarboxy-diphenyl diselenide, 4,4'-bischlorodiphenyl diselenide, 2,4,6,2',4',6'-hexamethyldiphenyl diselenide) could be substrates of rat brain TrxR. In the presence of partially purified rat brain TrxR, dipheny...
Waste cooking oil can be recovered and processed into animal feed additives after purification. Unfortunately, the traditional purification processes are insufficient for the removal of the harmful compounds formed during frying, mainly about malondialdehyde and other 2-thiobarbituric acid reactive substances. In the present paper, firstly, a simple and reliable HPLC method was developed to measure the content of malondialdehyde in purified waste cooking oil. The detection limit and the standard recovery of the analysis method are 1.20x10-5gl-1 and 96.5-99.2%, respectively, which is accurate and valid enough for the detection of malondialdehyde in waste cooking oil. Furthermore, the removal of malondialdehyde and other 2-thiobarbituric acid reactive substances from waste cooking oil was in...
A galactose-specific seed lectin was purified from the legume Spatholobus parviflorus and crystallized using the hanging-drop vapour-diffusion technique. Thecrystals belonged to space group P1, with unit-cell parameters a = 60.998, b=60.792, c = 78.179-, = 101.32, = 91.38, = 104.32. X-ray diffraction data were collected under cryoconditions (100-K) to a resolution of 2.04- using a MAR image-plate detector system mounted on a rotating-anode X-ray (Cu-K) generator. Molecular replacement using legume-lectin coordinates as a search model gave a tetrameric structure.
Phage display is effective in screening peptides that mimic venom's neutralizing epitopes. A phage display cyclized heptapeptide library (C7C library) was panned with purified divalent antivenin IgG, which neutralizes Naja naja atra venom (NAV) and Bungarus multicinctus venom (BMV). The selected heptapeptide sequences were aligned with known protein sequences of NAV and BMV in GenBank. One of the four consensus sequences, L/PKSSLL, mimicked the crucial epitope on Loop III of Taiwan cobra cardiotoxin that is associated with the venom's lethal potency. In dot blot analysis, several clones showed varying reactivities for NAV monovalent antivenin and lesser cross-reactions with BMV monovalent antivenin. The KSSLLRN-carrying phage occurred four times in selected clones and showed the strongest ...
Salt stress elicits betaine accumulation to high levels in species from several diverse dicot families (Chenopodiaceae, Amaranthaceae, Convolvulaceae, Solanaceae, and Asteraceae). FAM-MS studies with deuterated precursors showed that species from all these families synthesize betaine from choline. Enzyme assays and immunotitration data showed that all accumulating species contained betaine aldehyde dehydrogenase (BADH) enzyme activity recognized by antibodies raised against purified BADH isolated from Spinacia oleracea. Immunoblotting indicated that the BADH monomer was in all cases of Mr {approx} 63,000. The similarity of BADH in the different species is consistent with a single evolutionary origin for the betaine pathway. This was supported by the presence in immunoblots of a cross-reacting band at Mr {approx} 63,000 in Magnolia x Soulangiana, a primitive angiosperm.
Cholera toxin catalyzes transfer of radiolabel from ["3"2P]NAD"+ to several peptides in particulate preparations of human foreskin fibroblasts. Resolution of these peptides by two-dimensional gel electrophoresis allowed identification of two peptides of M/sub r/ = 42,000 and 52,000 as peptide subunits of a regulatory component of adenylate cyclase. The radiolabeling of another group of peptides (M/sub r/ = 50,000 to 65,000) suggested that cholera toxin could catalyze ADP-ribosylation of cytoskeletal proteins. This suggestion was confirmed by showing that incubation with cholera toxin and ["3"2P]NAD"+ caused radiolabeling of purified microtubule and intermediate filament proteins.
Thirty bacterial isolates were obtained from different sources and sites at Jeddah, Saudi Arabia, on chitin agar medium; 9 of the 30 isolates were cultured in liquid medium containing chitin as sole carbon and nitrogen sources. Isolate SM21, which was isolated from shrimp shells, showed the best growth and chitinase production in liquid medium. According to its morphological, physiological and biochemical characteristics, SM21 belongs to the genus Streptomyces and was identified as Streptomyces anulatus SM21. Identification was confirmed using 16S rDNA analysis. The chitinase enzyme was precipitated with 80% NH4SO4 and purified using DEAE-cellulose ion exchange chromatography followed by Sephadex G-100 gel filtration. The molecular weight determined using sodium dodecyl sulfate polyacrylam...
{sup 90}Sr, with a long half-life of 28.5 years, is the most dangerous strontium isotope. The adsorption behavior of radionuclides in the environment are closely related to the safe disposal of radioactive wastes. Since various types of minerals may exist in and around the repositories used for ultimate disposal of nuclear waste, the adsorption behavior of certain radionuclides onto and from these minerals and similar adsorbents should be studied in order to estimate the rates of transport of the nuclides in the event of water penetration into and through the repository. Information on the adsorption properties of the purified individual clay minerals may not be sufficient to predict the adsorption properties of a corresponding mixture, because these clay minerals may interact with each other and lead to modification of the adsorption properties of the mixture as compared to the pure minerals. The adsorption behavior of strontium on kaolinite and montmorillonite ...
A new microbial transglutaminase (MTGase or MTG, EC 2.3.2.13) from a Streptomyces sp. strain isolated from Brazilian soil samples was characterized in crude and purified forms. The aim of this work is to provide relevant information about a new transglutaminase and to compare its characteristics with the well-known commercial transglutaminase from Ajinomoto Co. Inc. (Activa? TG-BP). The enzyme from Streptomyces sp., in both crude and pure forms, exhibited optimal activity in the 6.0?6.5 pH range and at 35?40?C. The results for the commercial enzyme were the same. A second maximum of activity was observed at pH?10.0 with both the crude Streptomyces sp. enzyme and the commercial enzyme. This interesting fact has not been reported in the literature previously. The fact that this second maximu...
A rickettsia related to but distinct from the spotted fever agent, Rickettsia rickettsii, has been detected in 167 (18.9%) of 884 Rhipicephalus sanguineus taken off dogs in central and northern Mississippi. The organisms could readily be isolated in male meadow voles (Microtus pennsylvanicus), where it produced massive infections in the tissues of tunica vaginalis. It was practically nonpathogenic for male guinea pigs, although inoculation of these animals with infected tunica vaginalis of voles afforded in 30 of 38 instances solid immunity to challenge with virulent R. rickettsii. The Rhipicephalus rickettsia grew well in monolayers of chicken embryo fibroblast, Vero, mouse L, and HeLa cells. Cytopathogenic effects were minimal unless large concentrations of rickettsiae were used as inocula. It also could be established in embryonated hen eggs but only after injection of massive doses of L cell-propagated organisms. Serological tests (complement fixation, microagglutination and/or ...
We evaluated the use of a novel gene porter (Den123-a nontoxic self-assembled dendritic spheroidal nanoparticle made of biodegradable monomers), aiming to enhance and improve the desired immune response in protection from allergy. Footpad DNA immunization in Balb/c mice was done three times using the Bet v 1a gene with or without Den123 with 2-week intervals followed by sensitization with rBetv1 (5mg) in alum twice in a weekly interval. Different doses of pCMV-Betv1 were used (10mg and 100mg). The protective role of different formulations was evaluated by measuring the IgG1, IgG2a and IgE antibody production, cytokine release of isolated splenocytes and b-hexosaminidase release from the RBL cells. Higher and increasing ratios of IgG2a/IgG1 were seen in mice which received plasmids in combi...
Xenotropic murine leukemia virus-related virus (XMRV) was discovered in human prostate tumors and later in some chronic fatigue syndrome (CFS) patients. However, subsequent studies have identified various sources of potential contamination with XMRV and other murine leukemia virus (MLV)-related sequences in test samples. Biological and nucleotide sequence analysis indicates that XMRV is distinct from known xenotropic MLVs and has a broad host range and cell tropism including human cells. Therefore, it is prudent to minimize the risk of human exposure to infection by evaluating XMRV contamination in cell lines handled in laboratory research and particularly those used in the manufacture of biological products. Nested DNA PCR assays were optimized for investigating XMRV gag and env sequences in various cell lines, which included MRC-5, Vero, HEK-293, MDCK, HeLa, and A549, that may be used in the development of some vaccines and other cell lines broadly used in ...
OBJECTIVES: The authors examine the rationale and efficacy of pre-travel hepatitis B immunization strategies based on itinerary and presumed on-travel risk behaviour. METHODS: A large survey among 26,640 Danes survey provided data on journey lengths and destinations, immunization coverage, risk behaviour and knowledge. RESULTS: The estimated cumulative lifetime stay in endemic areas outside Europe is 4.3 months. The majority of risk situations are involuntary and unforeseeable. The majority of risk situations occur on short-term journeys. 5% nonimmune and 5% short-term travellers experienced at least one risk situation such as injections/operations/tattoos on their journey. The level of knowledge of hepatitis A and B is low. CONCLUSIONS: The rationale and efficacy of current immunization strategies are challenged. Based on the results presented here and the availability of vaccines with long-lasting coverage, the authors find that recommendations of immunization ...
1. Gamma globulin metabolism and distribution were studied employing rabbit gamma globulin (RGG)I(131) 24 times in 13 control rabbits. Similar studies were performed before and during the ananmestic response in 4 rabbits previously sensitized with a polyvalent pneumococcal vaccine. 2. During the ananmestic response, gamma-globulin levels increased from 1.0 to 6.0 gm/100 ml, and the gamma-globulin pool increased from 0.7 to 4.7 gm/kg. There was no change in the intravascular-extravascular partition of gamma globulin. 3. Gamma globulin degradation increased from 0.06 to 0.33 gm/kg/day during the 28 days of the immunization period while gamma globulin synthesis increased even further to average 0.47 gm/kg/day. Following the attainment of elevated gamma globulin levels the fractional rate of RGG-I(131) turnover increased from 8.0 to 12.5 per cent/day. 4. No differences were noted in the metabolism of homologous or autologous gamma globulin regardless of the allotypic ...
Hematopoietic stem cells (HSCs) have remained at the forefront of stem cell research for the past 50 years, since the therapeutic potential of bone marrow transplantation was realized. Uniquely, among stem and progenitor cells, research progress has been made in parallel between the laboratory benchtop and hospital bedside during this period. Integral to this work has been the role of the transfusion medicine services in the collection, storage, and processing of HSCs. The next decade promises to bring further developments: with new fields of cellular therapies, stem cell vaccination, and stem cell drug testing opening up. This article summarizes exciting areas of research concerning the behavior and potential clinical applications of HSCs. For the purposes of clarity, we describe in turn the trafficking and transfer of HSCs; ex vivo expansion of HSC units from different sources; and finally, applications of specifically selected subsets of hematopoietic cells and ...
Global meat and milk consumption is exponentially increasing due to population growth, urbanization and changes in lifestyle in the developing world. This is an excellent opportunity for developing countries to improve the livestock sector by using technological advances. Biotechnology is one of the avenues for improved production in the "Livestock revolution". Biotechnology developments applied to livestock health, nutrition, breeding and reproduction are improving with a reasonable pace in developing countries. Simple bio-techniques such as artificial insemination have been well implemented in many parts of the developing world. However, advanced technologies including transgenic plant vaccines, marker assisted selection, solid state fermentation for the production of fibrolytic enzymes, transgenic fodders, embryo transfer and animal cloning are confined largely to research organizations. Some developing countries such as Taiwan, China and Brazil have considered ...
Summary Background Treatment of localised renal cell carcinoma consists of partial or radical nephrectomy. A substantial proportion of patients are at risk for recurrence because no effective adjuvant therapy exists. We investigated the use of an autologous, tumour-derived heat-shock protein (glycoprotein 96)-peptide complex (HSPPC-96; vitespen) as adjuvant treatment in patients at high risk of recurrence after resection of locally advanced renal cell carcinoma. Methods In this open-label trial, patients were randomly assigned to receive either vitespen (n=409) or observation alone (n=409) after nephrectomy. Randomisation was done in a one to one ratio by a computer-generated pseudo-random number generator, with a block size of four, and was stratified by performance score, lymph node stat...
A factor which agglutinated the spores of Ceratocystis fimbriata in the presence of Ca2+ was purified from sweet potato (Ipomea batatas Lam cv. Norin[1]) root. Element composition of the purified factor was as follows; analysis found: C (29.8%), H (3.97%), O (65.34%), N (0.81%): calculated for C43H69O70N1: C (30.02%), H (4.01%), O (65.15%), N (0.81%). The factor was mainly composed of galacturonic acid (53% of dry weight) and contained arabinose, fucose, and unidentified component as minor components. The factor also agglutinated A-, B-, AB-, and O types of human erythrocytes to almost the same degree in the presence of Ca2+. The differential spore-agglutinating activity of the factor depended on the pH of the assay medium; it agglutinated similarly the germinated spores of sweet potato and coffee strains at pH 7.5 and 5.5, whereas it displayed a distinct differential agglutinating activity at pH 6.5. The factor was assayed for ...
The availability of polysilicon feedstock has become a major issue for the photovoltaic (PV) industry in recent years. Most of the current polysilicon feedstock is derived from rejected material from the semiconductor industry. However, the reject material can become scarce and more expensive during periods of expansion in the integrated-circuit industry. Continued rapid expansion of the PV crystalline-silicon industry will eventually require a dedicated supply of polysilicon feedstock to produce solar cells at lower costs. The photovoltaic industry can accept a lower purity polysilicon feedstock (solar-grade) compared to the semiconductor industry. The purity requirements and potential production techniques for solar-grade polysilicon have been reviewed. One interesting process from previous research involves reactive gas blowing of the molten silicon charge. As an example, Dosaj et all reported a reduction of metal and boron impurities from silicon melts using reactive gas blowing ...
Full text: The maturation of chicken forebrain is protracted and occurs well after synapse formation providing a good model for studying mechanisms of brain maturation. Using microslices from immature (10 day) and adult chicken forebrain prepared after decapitation, we have examined functional properties of NMDA and AMPA receptors by measuring agonist-induced uptake of "4"5Ca"2"+ . The rate and extent of NMDA induced "4"5Ca"2"+ accumulation decreased during maturation with no change in EC_5_0. The rate and extent of the AMPA induced response also decreased with a 60-fold increase in EC_5_0. However, the total NMDA receptor content did not change as indicated by 3 H-MK801 binding and NR1 immunoreactivity in P2 fractions. Similarly, there was no change in the B_m_a_x of "3H-AMPA, though there was a two-fold increase in K_D, and little or no change in the immunoreactivity in GluR1, 2, 2/3 or 4. These results suggest that it is the regulation of receptors, their subunit composition and/or ...
Thromboxane A2, the predominant product of arachidonic acid metabolism in the blood platelet, is a potent vasoconstrictor and platelet agonist. During its biosynthesis from cyclic endoperoxide, 12(S)-hydroxy-5Z,8E,10E-heptadecatrienoic acid (HHT) is formed in equal amounts. The further metabolism of HHT, catalyzed by 15-hydroxyprostaglandin dehydrogenase, leads to 12-oxo-5Z,8E,10E-heptadecatrienoic acid (Oxo-HT). Sample workup procedures are described which allow for the sensitive and reproducible determination of these two arachidonic acid metabolites in human plasma by gas chromatography-mass spectrometry in the presence of deuterated analogues as internal standards. HHT is derivatized to the pentafluorobenzyl ester tert-butyldimethylsilyl ether. In order to enable quantification of low concentrations of about 10 pg/ml in nonstimulated human plasma, the samples have to be purified by HPLC. Oxo-HT is derivatized to the pentafluorobenzyl ester, which is ...
The effects of purified Helminthosporium maydis T (HmT) toxin on active Ca/sup 2 +/ transport into isolated mitochondria and microsomal vesicles were compared for a susceptible (T) and a resistant (N) strain of corn (Zea mays). ATP, malate, NADH, or succinate could drive /sup 45/Ca/sup 2 +/ transport into mitochondria of corn roots. Ca/sup 2 +/ uptake was dependent on the proton electrochemical gradient generated by the redox substrates or the reversible ATP synthetase, as oligomycin inhibited ATP-driven CA/sup 2 +/ uptake while KCN inhibited transport driven by the redox substrates. Purified native HmT toxin completely inhibited Ca/sup 2 +/ transport into T mitochondria at 5 to 10 nanograms per milliliter while transport into N mitochondria was decreased slightly by 100 nanograms per milliliter toxin. Malate-driven Ca/sup 2 +/ transport in T mitochondria was frequently more inhibited by 5 nanograms per milliliter toxin than succinate or ...
In mice, coat pigmentation requires a stem cell (SC) system in which the survival, proliferation, and differentiation of melanocytes (MCs) are regulated by microenvironments in hair follicles (HFs). In vitro systems are required to analyze the behavior of single melanocyte stem cells (MCSCs) and their potential to form SC systems in vivo. We describe here an experimental system for the isolation, self-renewal, and differentiation of MCSCs, as well as an in vivo reconstitution assay for assessing their potential. Using Dct(tm1(Cre)Bee)/CAG-CAT-GFP mice, we show that, in the presence of stem cell factor and basic fibroblast growth factor and the XB2 feeder cell line, purified MCSCs can undergo clonogenic proliferation, resulting in c-Kit(low) side scatter(low) cells. In culture, these cells maintain their capacity to differentiate and reconstitute an MCSC system in HFs. As these cells are present in the upper part of the HF near the bulge region, express only low ...
A hyaluronic acid binding fraction was purified from the supernatant media of both 3T3 and murine sarcoma virus (MSV) transformed 3T3 cultures by hyaluronate and immunoaffinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis resolved the hyaluronate affinity-purified fraction into three major protein bands of estimated molecular weight (M/sub r,e/) 70K, 66K, and 56K which contained hyaluronate binding activity and which were termed hyaluronate binding proteins (HABP). Hyaluronate affinity chromatography combined with immunoaffinity chromatography, using antibody directed against the larger HABP, allowed a 20-fold purification of HABP. Fractions isolated from 3T3 supernatant medium also contained additional binding molecules in the molecular weight range of 20K. This material was present in vanishingly small amounts and was not detected with a silver stain or with (/sup 35/S)methionine label. The three protein species ...
A factor produced by the plerocercoid stage of S. mansonoides mimics some, but not all, of the actions reported for hGH. The biological actions of plerocercoid growth factor (PGF) suggest structural similarity to human GH (hGH). Plerocercoid membranes were solubilized, and PGF was purified more than 1000-fold by hGH receptor affinity chromatography. The ability of purified PGF to displace [125I]hGH from monoclonal antibodies specific for four distinct nonoverlapping antigenic determinants of hGH and from an anti-hGH polyclonal antibody was tested in liquid phase RIA. All of the hGH antibodies cross-reacted with PGF, with potencies ranging from more than 60% to less than 1% that of the hGH standard. Of the four major epitopes of hGH defined by the monoclonal antibodies used in this study, only one is not represented to a significant extent in PGF. The epitope of hGH that is only marginally present in PGF is highly conformationally dependent, and ...
The effects of gamma irradiation at 0.2#approx#0.4 Mrad and condition for processing of buckwheat noodles on sensory quality and physical properties were investigated. The results were as follows: (1) Sensory evaluation of buckwheat noodles, prepared with the 70 % ratio of buckwheat flour, was decreased according to an increase of irradiation dose of buckwheat flour. Hardness and adhesiveness of buckwheat noodles caused by irradiation should be given large effect to the texture. Elasticity of buckwheat noodles was linearly decreased with increasing of irradiation dose of buckwheat flour, and elasticity of buckwheat noodles was decreased about 15 % by an increase of irradiation dose at 0.3 Mrad. (2) Maximum torque in Farinograph test was linearly decreased with increasing of irradiation dose of buckwheat flour, and maximum torque was decreased about 3 % by increasing of irradiation dose with 0.1 Mrad. (3) Sensory evaluation was also decreased by increasing content of irradiated buck ...
The objective of this research is to determine the role of DNA repair in mutagenesis and carcinogenesis in mammalian cells. Use of the host-cell reactivation viral suicide enrichment procedure was initiated in the isolation of repair-deficient mutants. Lightly mutagenized BHK cells were infected with irradiated Herpes simplex virus (HSV); several radiation-sensitive strains were isolated among the survivors of the infection. The characterization of these strains is progressing and the enrichments are continuing. That alterations in the frequency of mutation of C3H/10T 1/2 cells, occurring as a result of holding the cells in a confluent state following treatment with ethylmethane sulfonate, parallel the alterations in the frequency of neoplastic transformation was found. The repair capabilities of BHK cells were found to be intermediate in comparison to repair-proficient and -deficient human cells with regard to the reactivation of HSV treated with various ...
The epileptogenic and neurodegenerative effects of gamma-dendrotoxin, from Dendroaspis angusticeps, a specific blocker of a non-inactivating, voltage-sensitive K+ channel, were studied after focal injection into one dorsal hippocampus in rats pretreated with CGP040116, a N-methyl-D-aspartate (NMDA) receptor antagonist, and in rats bearing a monolateral surgical lesion of the Schaffer collaterals whose terminals originate from CA3 pyramids and release glutamate in the CA1 hippocampal area. Administration of 35 pmol gamma-dendrotoxin elicited in all of the treated animals (n=8) bilateral EEG discharges and damage to the hippocampal formation. Quantitation of the damage revealed significant bilateral neuronal cell loss in the CA1, CA3 and CA4 pyramidal cell layers. The lowest dose (0.35 pmol; n=4) of the toxin used did not affect EEG activity and failed to cause significant hippocampal cell loss whereas the 3.5 pmol (n=6) dose caused EEG seizures and hippocampal cell ...
The authors have found that heat-inactivated serum (57/sup 0/C for 1 hour) from normal chickens reduces the proliferation of mitogen-stimulated chicken and murine splenocytes as well as some transformed mammalian lymphoblastoid cell lines. Greater than a 50% reduction in /sup 3/H-thymidine incorporation was observed when concanavalin A (Con A)-activated chicken splenocytes that were cultured in the presence of 10% autologous or heterologous serum were compared to mitogen-stimulated cells cultured in the absence of serum. Normal chicken serum (10%) also caused greater than 95% suppression of /sup 3/H-thymidine incorporation by bovine (EBL-1 and BL-3) and gibbon ape (MLA 144) transformed lymphoblastoid cell lines. The only cell line tested that was not inhibited by chicken serum was an IL-2-dependent, murine cell line. Chicken serum also inhibited both /sup 3/H-thymidine incorporation and IL-2 synthesis by Con A-activated murine splenocytes. Suppression was caused by ...
A nucleoside triphosphate binding site on calf thymus RNA polymerase II was identified by using photoaffinity analogues of adenosine 5'-triphosphate and guanosine 5'-triphosphate. Both radiolabeled 8-azidoadenosine 5'-triphosphate (8-N3ATP) and radiolabeled 8-azidoguanosine 5'-triphosphate (8-N3GTP) bound to a single polypeptide of this enzyme. This polypeptide has a molecular mass of 37 kilodaltons and an isoelectric point of 5.4. Ultraviolet (UV) irradiation was necessary for photolabeling to occur. In addition, no labeling occurred when the probe was prephotolyzed or when the enzyme was inactivated. Furthermore, photolabeling of the enzyme could be decreased by preincubation with natural substrates. To provide evidence that the radiolabeled polypeptide forms a part of the domain of the nucleoside triphosphate binding site, experiments were performed using unlabeled 8-N3ATP. Although this unlabeled analogue was not a substrate ...
Glucoamylase and pullulanase were immobilized on reconstituted bovine-hide collagen membranes using the covalent azide linkage method. A pretanning step was incorporated into the immobilization procedure to enable the support matrix to resist proteolytic activity while accommodating an operating temperature of 50 degrees C. The immobilized glucoamylase and pullulanase activities were 0.91 and 0.022 mg dextrose equivalent (DE) min(-1) cm(-2) of membrane, respectively. Immobilized glucoamylase had a half-life of 50 days while the immobilized pullulanase had a half-life of 7 days. This is a considerably improved stability over that reported by other researchers. The enzymes were studied in their free and immobilized forms on a variety of starch substrates including waxy maize, a material which contains 80% alpha-1-6-glucosidic linkages. Substrate concentrations ranged from 1% to a typical commercial concentration of 30%. Conversion efficiencies of 90-92% DE were obtained with free and ...
Burnaid is a sorbalene-based cream containing 40 mg/g of tea tree oil and 1 mg/g of triclosan. This investigation was carried out to determine the effect of Burnaid, a commercial tea tree oil preparation, against Enterococcus faecalis (ATCC29212), Staphylococcus aureus (ATCC29213), Escherichia coli (ATCC25922), and Pseudomonas aeruginosa (ATCC27853), with the activity of the base product in the commercial preparation. The organisms were suspended in sterile saline (0.5 McFarland Standard) and inoculated onto horse blood agar (E. faecalis and S. aureus) or Mueller-Hinton agar (E. coli and P. aeruginosa). One hundred microliters of Burnaid unsterilized, Burnaid sterilized and the base product (Tinasolve) were placed in duplicate in wells cut into the agar plates. Sterility and inactivation cultures were also performed on the samples. None of the samples were found to be contaminated with bacteria prior to testing. Only S. aureus and E. coli showed zones of growth ...
In this study, the biosorption of Cd(II), Ni(II) and Pb(II) on Aspergillus niger in a batch system was investigated, and optimal condition determined by means of central composite design (CCD) under response surface methodology (RSM). Biomass inactivated by heat and pretreated by alkali solution was used in the determination of optimal conditions. The effect of initial solution pH, biomass dose and initial ion concentration on the removal efficiency of metal ions by A. niger was optimized using a design of experiment (DOE) method. Experimental results indicated that the optimal conditions for biosorption were 5.22 g/L, 89.93 mg/L and 6.01 for biomass dose, initial ion concentration and solution pH, respectively. Enhancement of metal biosorption capacity of the dried biomass by pretreatment with sodium hydroxide was observed. Maximal removal efficiencies for Cd(II), Ni(III) and Pb(II) ions of 98, 80 and 99% were achieved, respectively. The biosorption capacity of A. ...
beta-Bungarotoxin (beta-Bgt), a presynaptic phospholipase A(2) (PLA(2)) neurotoxin isolated from the venom of Bungarus multicinctus, consists of A chain and B chain. The goal of the present study is to explore the functional contribution of the two subunits to the toxicity of beta-Bgt. beta-Bgt was found to induce apoptotic death of SK-N-SH cells via elevating intracellular Ca(2+) and intracellular ROS production. Moreover, an activation of p38 MAPK was associated with the cytotoxicity of beta-Bgt. SB202190 (p38 MAPK inhibitor), N-acetylcysteine (antioxidant reagent), 1,2-bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid (BAPTA) (Ca(2+) chelator) and the inhibitors of Ca(2+) release from intracellular depots (ruthenium red and 2-aminoethoxydiphenyl borate) effectively attenuated the cytotoxicity of beta-Bgt. In sharp contrast to the inability of A chain, B chain was able to induce cytotoxic effects on SK-N-SH cells as beta-Bgt did. Abolishment of PLA(2) activity did not significantly ...
CNPT-3 is the laboratory designation of a strain of bacteria that reproduces best at deep-sea pressures. It was isolated from a sample collected at a 5800 m depth of the Pacific Ocean. This isolation was achieved from a sample that had been retrieved without warning. This year we demonstrated that there are similarly behaving bacteria that can be isolated from samples even when retrieved with decompression. The purpose of this project was to define the temperatures and pressures over which CNPT-3 can exist and function. (1) At 2"0C CNPT-3 is eurybathic from about 1000 to 6500 m, as judged from an only slightly pressure dependent generation time. (2) CNPT-3 is unquestionably barophilic at 2"0C, with doubling times of 9 hours at 580 bars and over 18 hours at 1 bar. (3) At 10"0C, reproduction is adversely affected at 1 bar to the extent that CNPT-3 becomes obligately barophilic at 10"0C. Yet reproduction is enhanced above 280 bars and occurs with a regeneration time of 4 hours at 690 bars ...
The presence of endothelin (ET)-like immunoreactivity and the cardiovascular effects of mammalian ET-1 in fish have been reported. To identify ET-related peptides in fish, we screened the cDNA library of the salmon (Oncorhynchus keta) stomach by means of rapid amplification of cDNA ends, and we cloned cDNAs encoding an ET-related peptide. The salmon ET-related sequence of 21 amino acids is identical to the trout ET-1 peptide recently purified from kidney specimens of Oncorhynchus mykiss. The deduced amino acid sequence of salmon pre-proET-1 (PPET-1) comprises 244 amino acids, including a putative signal sequence and mature ET-1, as well as big ET-1 and ET-1-like sequences. This precursor, the first reported PPET-1 sequence for Salmoniformes, Teleostei, has low homology with the sequences of human, mouse, frog (Xenopus laevis), and zebrafish (Danio rerio) PPET-1 (26%, 29%, 24%, and 39%, respectively). PMID:16740985
Sulfated zirconia (SO4-ZrO2) catalysts, prepared with three different sulfur loading contents (0.75%, 1.8% and 2.5%) at two calcination temperatures (500degreeC and 700degreeC), were tested for use in the transesterification of purified palm oil (PPO) and the esterification of palm fatty acid (PFA) in near-critical and super-critical methanol. Techniques including BET, XRD, NH3- and CO2-TPD revealed that the sulfur content and calcination temperature strongly affects the catalyst base-acid site, specific surface area, average pore size, phase structure, and thus the catalytic reactivity. The most suitable sulfur loading content was found to be 1.8% and the optimum calcination temperature 500degreeC. The results show that the use of SO4-ZrO2 reduces esterification reaction times, the amount...
DNA polymerase gamma from purified nuclei of EMT-6 cells (mice) seems to be identical to the mitochondrial DNA polymerase from the same source following several criteria. These two enzyme activities are strongly inhibited by ethidium bromide and acriflavin, while proflavin, acridine orange, daunomycin and chloroquine inhibition is less pronounced. In the case of DNA polymerases alpha and beta very little inhibition by ethidium bromide was observed. Intercalation of this dye in a poly dA-dT 12-18 template-primer was studied spectrophotometrically under conditions similar to those in the in vitro DNA polymerase assay. The polymerase assay. The inhibition by this drug of the mitochondrial DNA polymerase gamma activity was shown to be competitive at varying concentrations of TTP while the inhibition was of the non-competitive type at different concentrations of poly dA-dT 12-18. We conclude that the drug, most probably in the intercalated form, is able to interact with ...
Novel complexes of the type (Pt(DACH)(N-R-iminodiacetate)), wherein DACH represents (R,S)- and (R,R)-1,2 diaminocyclohexane and R represents /minus/Me, /minus/EtOH, and /minus/CH/sub 2/Ph groups, have been prepared, purified, and characterized by spectroscopic techniques (/sup 1/H, /sup 13/C, and /sup 195/Pt NMR;MS(FAB);IR) and by the measurement of selected physical properties (pH, pK/sub a/, conductivity, and molecular weights). The data are consistent with the formation of two diastereomeric complexes in unequal proportions in which the N-R-iminodiacetate ligand appears to be bonded as a pseudofacial tridentate chelate. One are of the ligand forms a stable five-membered-ring O,N-chelate while the other arm appears to be involved in ion-pair formation (zwitterion-like) involving the carboxylate anion and the formally positive Pt(II) central metal atom. It has been demonstrated indirectly that an active impurity was present in predictably inactive bulk complexes ...
Somatogenic hormones play an important role in regulation of receptors for prolactin (PRL) and estrogen. Plerocercoids of the tapeworm, S. mansonoides produce a factor which mimics some, but not all of the actions reported for GH. Intact female rats were subjected to a constant infusion of plerocercoid growth factor (PGF) via a subcutaneous infection for two weeks to determine if PGF influences receptors for PRL, GH or estradiol. The rate of weight gain in the PGF-treated rats was accelerated in spite of a marked reduction in serum GH. Partially-purified PGF specifically displaced [125I]hGH from rat liver receptors but microsomes prepared from rats treated with PGF specifically bound significantly less [125I]hGH than microsomes from control rats. The reduction in [125I]hGH binding was not due to occupancy or to a change in affinity but to a suppression in receptor concentration. Scatchard analysis of [3H]estradiol binding in rat liver cytosols shows a 50% reduction ...
The non-blast furnace coke produced in the shaft ovens of the Angarsk Semi-coking Plant is used principally in gas generator ovens and partially as a carbonaceous reducing agent in the ferroalloy process. The sulfur content of this coke is not limited by the standards documents, but it is significant, especially in the production of process gases. In a number of recent years the sulfur removal system of the gasification divisions have been experiencing difficulties, not achieving the assigned gas purification parameters, with a resulting H/sub 2/S concentration in the purified gas of 300-700 mg/m/sup 3/ as compared to the standard of 50. Realizing that in the gasification practically all the organic matter of the coke, including the sulfur, enters the gas, the impairment in the quality of the sulfur removal is related to the increase in the sulfur content of the coke. Thus, an analysis of the results was conducted for the determination of the sulfur content of the ...
We have used a cell-free polymerase I transcription system derived from HeLa cells to study the regulation of human rRNA synthesis. Analysis of deletion mutants spanning the start site of transcription at nucleotide +1 indicates that the control region affecting initiation of human rRNA synthesis is contained within sequences from nucleotides -158 to +18. This promoter region can be subdivided into (i) a central segment of approximately 40 base pair that is required for transcription and (ii) flanking sequences that influence the efficiency of transcription in vitro. We have examined the in vitro transcriptional activity of the human extract under various conditions that are thought to modulate rRNA synthesis in vivo. Cell-free extracts prepared from HeLa cells infected with adenovirus 2 synthesize human rRNA at levels greatly decreased relative to uninfected cell extracts. By contrast, in vitro transcription of human rRNA is stimulated 2- to 3-fold by the addition of ...
By improving the currently used lactoperoxidase method, we were able to obtain radioiodinated chicken luteinizing hormone (LH) that shows high specific binding and low nonspecific binding to a crude plasma membrane fraction of testicular cells of the domestic fowl and the Japanese quail, and to the ovarian granulosa cells of the Japanese quail. The change we made from the original method consisted of (1) using chicken LH for radioiodination that was not only highly purified but also retained a high receptor binding potency; (2) controlling the level of incorporation of radioiodine into chicken LH molecules by employing a short reaction time and low temperature; and (3) fractionating radioiodinated chicken LH further by gel filtration using high-performance liquid chromatography. Specific radioactivity of the final {sup 125}I-labeled chicken LH preparation was 14 microCi/micrograms. When specific binding was 12-16%, nonspecific binding was as low as 2-4% in the ...
Four extracellular keratinases (designated KI, KII, KIII, and KIV) were produced during submerged aerobic cultivation in a medium containing native human foot skin (NHFS) for enzyme synthesis. The molecular weights, determined by SDS-PAGE, were 25, 50, 34, and 19kDa, respectively. Gel filtration of the four purified enzymes in native conditions indicated that active keratinase KI is a novel homo-octamer, KII a homo-dimer, and KIII and KIV monomers. All four keratinases exhibited high activities at pH 8.0-10.0 with an optimal pH of 9.0. The optimal temperature for keratinolytic activity of KI, KII, and KIII was approximately 50, and 60degreeC for KIV. One millimolar of PMSF completely inhibited the keratinolytic activities of the four enzymes. The N-terminal sequences of KI, KII, and KIII s...
Four proteins from NCBI cog1816, previously annotated as adenosine deaminases, have been subjected to structural and functional characterization. Pa0148 (Pseudomonas aeruginosa PAO1), AAur1117 (Arthrobacter aurescens TC1), Sgx9403e, and Sgx9403g have been purified and their substrate profiles determined. Adenosine is not a substrate for any of these enzymes. All of these proteins will deaminate adenine to produce hypoxanthine with k(cat)/K(m) values that exceed 10(5) M(-1) s(-1). These enzymes will also accept 6-chloropurine, 6-methoxypurine, N-6-methyladenine, and 2,6-diaminopurine as alternate substrates. X-ray structures of Pa0148 and AAur1117 have been determined and reveal nearly identical distorted (?/?)(8) barrels with a single zinc ion that is characteristic of members of the amidohydrolase superfamily. Structures of Pa0148 with adenine, 6-chloropurine, and hypoxanthine were also determined, thereby permitting identification of the residues responsible for ...
The most abundant minor hemoglobin component of human hemolysate is Hb A1c, which has glucose bound to the N-terminus of the beta chain by a ketoamine linkage. Hb A1c is formed slowly and continuously throughout the 120 day lifespan of the red cell. It can be synthesized in vitro by incubating purified hemoglobin with 14C-glucose. Other minor components, Hb A1a1 and Hb A1a2 are adducts of sugar phosphates at the N-terminus of the beta chain. Hb A1b contains an unidentified nonphosphorylated sugar at the beta N-terminus. In addition, a significant portion of the major hemoglobin component (Hb Ao) is also glycosylated by a glucose ketoamine linkage at other sites on the molecule, including the N-terminus of the alpha chain and the epsilon-amino group of several lysine residues on both the alpha and the beta chains. The results indicate that the interaction of glucose and hemoglobin is rather nonspecific and suggests that other proteins are modified in a similar ...
Atomic Energy of Canada Limited (AECL) operates the Waste Treatment Centre (WTC) to treat and immobilize some of the low- level radioactive waste (LLRW) streams at the Chalk River Laboratories (CRL). The WTC at treats low- level radioactive liquid waste by removing the contaminants from the wastewater, concentrating them, and immobilizing them. The fundamental design concept for the WTC is to process the waste streams using forced circulation type liquid waste evaporation (LWE), to solidify the concentrates using thin film evaporator and to discharge the purified effluent into the Ottawa River following verification monitoring. The solidified product drums are stored in existing storage facilities in the CRL. The LWE was installed in the WTC to treat the LLRW. After about four (4) years of design, construction and cold commissioning, the active commissioning of the evaporator process using radioactive waste streams commenced in February 2000. The LWE has overcome ...
Atomic Energy of Canada Limited (AECL) operates the Waste Treatment Centre (WTC) to treat and immobilize some of the low- level radioactive waste (LLRW) streams at the Chalk River Laboratories (CRL). The WTC at treats low- level radioactive liquid waste by removing the contaminants from the wastewater, concentrating them, and immobilizing them. The fundamental design concept for the WTC is to process the waste streams using forced circulation type liquid waste evaporation (LWE), to solidify the concentrates using thin film evaporator and to discharge the purified effluent into the Ottawa River following verification monitoring. The solidified product drums are stored in existing storage facilities in the CRL. The LWE was installed in the WTC to treat the LLRW. After about four (4) years of design, construction and cold commissioning, the active commissioning of the evaporator process using radioactive waste streams commenced in February 2000. The LWE has overcome ...
Measurements of the Kapitza conductance to liquid helium II across the (100) surface of single crystals of copper are presented. The temperature range of these measurements was 1.6-- 2.1 K. The sample surfaces were subjected to several different treatments. Some surfaces were cleaned by low-energy argon ion bombardment, annealed in an ultrahigh-vacuum system, and preserved under vacuum until purified liquid helium was admitted. Other surfaces were intentionally damaged by machining and/or exposure to the atmosphere. The conductance after these latter treatments was found to be about a factor of three higher than that of the more ideally cleaned and annealed surfaces, and a significant difference in the temperature dependence of the conductance was also observed. Conductances were reproducible for similarly treated surfaces and correlated with surface damage determined by x-ray diffraction. The relationship of these results to the numerous current theories of the ...
In this study, two different humic acids extracted from England and China peat soil, as well as three purified products extracted from commercially available humic acids [Fluka (Fluka, Neu-Ulm, Germany, Alfa (Alfa Aesar, Ward Hill, USA) and Aldrich (Aldrich-Chemical Co, Gillingham-Dorset)] were obtained respectively with the modified IHSS (International humic substances society) procedure. The chemical and structural information of these humic acids were characterized by some techniques, such as elemental analysis, ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FTIR) and electropotential titration. UV-Vis spectra, infrared spectra, values for elemental composition and the contents of carboxyls, phenolic hydroxyls and total acidities of these humic acids were acquired. It is indicated that all these humic acids are different in the aspect of chemical structure, respectively, and these humic acids would be used for the ...
Gel filtration of human erythrocyte (RBC) lysate incubated with labeled thyroxine (Tu) or triiodothyronine (Tt) revealed co-elution of a major iodothyronine-binding fraction (R-2) and hemoglobin. Solutions of purified human hemoglobin and Tt also showed co-elution of hormone and hemoglobin. Because hematin and protoporphyrin were shown to bind labeled Tt, the oxygen-binding site on hemoglobin was excluded as the site of iodothyronine-hemoglobin interaction. Analysis of hormone binding by heme and globin moieties showed Tt binding to be limited to the heme fraction. Addition of excess unlabeled Tt to hemoglobin or heme incubated with labeled Tt indicated 75% to 90% of hormone binding was poorly dissociable. These observations suggested that the presence of hemoglobin in RBC lysate or in serum could influence the measurement of Tu and Tt by specific radioimmunoassay (RIA). Subsequent studies of the addition to serum of human hemoglobin revealed a significant ...
The effect of nutrition on reproductive performance was studied in a series of experiments using two models. Increased supply of nutrients was based on feeding lupin grain as a supplement and measurements of testicular growth or ovulation rates were used to measure reproductive characteristics. Purified sources of protein and energy were provided to determine the macronutrient(s), supplied with lupin feeding, responsible for initiating increases in testicular growth and ovulation rate. Increases in both of these parameters were closely related to the supply of additional energy but not protein. The production of acetate and glucose resulting from supplementary feeding with 750 g lupins/d was measured using infusions of "1"4C tracers. Subsequently, infusions of glucose and acetate (individually and in combination) were found to produce similar increases in ovulation rate to those measured in ewes receiving lupins. It was concluded that effects of nutrition on ...
The objective of this work was to investigate the efficiency of imidazole derivatives for corrosion inhibition of copper in 0.5 M hydrochloric acid. Corrosion inhibition was studied using impedance spectroscopy. Imidazole and its derivatives 4-methylimida-zole, 4-methyl-5-hydroxymethylimidazole, 1-phenyl-4-methylimidazole, 1-(p-tolyl)-4-methylimidazole were investigated. These studies have shown that 1-(p-tolyl)-4-methylimidazole is the best inhibitor in this series and that it acts as mixed inhibitor. The nature of the chemical interaction between these molecules and the copper surface was investigated by Cu exposed to solutions having two very different pH values: 0.5 M HCl and unbuffered purified water. X-ray photoelectron spectroscopy and secondary ion mass spectrometry were used to explore the nature of the interaction. Possible mechanisms of corrosion inhibition for these molecules are discussed.
A laminated material composed of glass cloth/polyimide film/epoxy resin will be used as an insulating material for superconducting coil of International Thermonuclear Experimental Reactor (ITER). In order to keep safe and stable operation of the superconducting coil system, it is indispensable to evaluate radiation resistance of the material, because the material is exposed to severe environments such as high radiation field and low temperature of 4 K. Especially, it is important to estimate the amount of gases evolved from the insulating material by irradiation, because the gases affect on the purifying system of liquid helium in the superconducting coil system. In this work, the gas evolution from the laminated material by gamma ray irradiation at liquid nitrogen temperature (77 K) was investigated, and the difference of gas evolution behavior due to difference of composition in the epoxy resin was discussed. It was found that the main gases evolved from the ...
Functionalization of carbon nanotubes is very challenging for their applications. The paper here describes a new method to functionalize multi-walled carbon nanotubes (MWCNTs) as specific affinity adsorbents. MWCNTs were acid purified and pretreated with (3-aminopropyl)-triethoxysilane (APTES) in order to introduce abundant amino groups on the surface of MWCNTs. After the conversion of amino groups to carboxyl groups by succinic acid anhydride, MWCNTs were attached to protein A or aminodextran using 1-ethyl-3,3' (dimethylamion)-propylcarbodiimide as a biofunctional crosslinker. The incorporation of aminodextran as a spacer arm noticeably increased the binding capacity of the APTES-modified MWCNTs for protein A. The application of affinity MWCNTs for purification of immunoglobulin G was then evaluated. The affinity of MWCNTs with AMD spacer exhibited a high adsorption capacity of {approx}361 {mu}g IgG/mg MWCNT (wet basis). About 75% of bound IgG was eluted ...
The time of appearance of newly synthesized rRNAs and ribosomal proteins (r-proteins) in the ribosomes of pea cotyledons (Pisum sativum L.) during germination was investigated. The ribosomal fraction was isolated and analyzed according to the method of germination of the embryo in the presence of labeled precursors or after pulse labeling of the embryos at different stages of germination. For the identification of newly synthesized rRNAs in the ribosomes we estimated the relative stability of labeled RNAs to the action of RNase, the sedimentation rate, the ability to be methylated in vivo in the presence of (/sup 14/C)CH/sub 3/-methionine, and the localization in the subunits of dissociated ribosomes. The presence of newly synthesized r-proteins in the ribosomes was judged on the basis of the electrophoretic similarity in SDS-disc electrophoresis of labeled polypeptides of purified ribosome preparations and of genuine r-proteins, as well as according to the ...
Synthesis genes encoding the human #alpha#- and #beta#-globin polypeptides have been expressed from a single operon in Escherichia coli. The #alpha#- and #beta#-globin polypeptides associate into soluble tetramers, incorporate heme, and accumulate to >5% of the total cellular protein. Purified recombinant hemoglobin has the correct stoichiometry of #alpha#- and #beta#-globin chains and contains a full complement of heme. Each globin chain also contains an additional methionine as an extension to the amino terminus. The recombinant hemoglobin has a C_4 reversed-phase HPLC profile essentially identical to that of human hemoglobin A_0 and comigrates with hemoglobin A_0 on SDS/PAGE. The visible spectrum and oxygen affinity are similar to that of native human hemoglobin A_0. The authors have also expressed the #alpha#- and #beta#-globin genes separately and found that the expression of the #alpha#-globin gene alone results in a marked decrease in the accumulation of ...
It Peripheral blood progenitor enriched CD34+ cells (PBPC) are rather often used as stem cell background in cancer patients following high dose therapy. Keeping in mind that precursor dendritic cells (DCs) originate from haematopoietic progenitor cells, purified CD34+ cells might also serve as starting cells for ex-vivo production of DC. The aim of the present study is to develop a clinical grade procedure for ex-vivo production of DC derived from enriched CD34+ cells. Various concentrations of CD34+ cells were grown in gas-permeable Teflon bags with different serum-free and serum-containing media supplemented with GM-CSF, IL-4, TNF-a, SCF, Flt-3L and INF-a. Serum-free CellGroSCGM medium for 7 days followed by CellGroDC medium in 7 days gave equal results as serum-containing medium. Follow...
A comprehensive dataset of NDV genome sequences was evaluated using bioinformatics to characterize the evolutionary forces affecting NDV genomes. Despite evidence of recombination in most genes, only one event in the fusion gene of genotype V viruses produced evolutionarily viable progenies. The codon-associated rate of change for the six NDV proteins revealed that the highest rate of change occurred at the fusion protein. All proteins were under strong purifying (negative) selection; the fusion protein displayed the highest number of amino acids under positive selection. Regardless of the phylogenetic grouping or the level of virulence, the cleavage site motif was highly conserved implying that mutations at this site that result in changes of virulence may not be favored. The coding sequence of the fusion gene and the genomes of viruses from wild birds displayed higher yearly rates of change in virulent viruses than in viruses of low virulence, suggesting that an ...
Partially purified UDPgalactosyltransferase (EC 2.4.1.22) from bovine milk has been used to synthesize millimolar amounts of compounds such as Gal#beta#(1#->#4)Glc, Gal#beta#(1#->#4)GlcNAc-#beta#-hexanolamine, and Gal#beta#(1#->#4)-GlcNAc#beta#(1#->#4)GlcNAc. The same method has been used to prepare similar compounds containing "1"3C-enriched galactopyranosyl moieties. Gal#beta#(1#->#4)GlcNAc-#beta#-hexanolamine was also synthesized in a solid-phase system in which the GlcNAc-#beta#-hexanolamine glycoside was covalently linked to agarose beads. At pH 7.0 and at 1 to 5 mM Mn"2"+"+ the yields of the galactosyl saccharides are greater than 90% by using 10% excess of UDPGal donor. The use of a 90% enriched [1-"1"3C]galactosyl residue allowed the determination of the most abundant conformer about the galactopyranosyl-glycoside linkage by analysis of the carbon-carbon coupling constants from Cl to Gal to the C3', C4', and C5' of GlcNAc or Glc. 3 figures, 1 ...
Combustion scientists are primarily concerned with the fuels most often burned as energy sources (coal, petroleum products, and natural gas), with the goal of learning to burn them as efficiently, intensely, and cleanly as possible. Discovering those slight rearrangements of chemical bonds that together account for the net chemical transformation is the key to understanding how combustion proceeds. Once these reactions have been defined, the chemist can determine the rate coefficient of each reaction as a function of temperature and assemble the information into flame models. The computer programs that use these models to predict experimental results combine two sets of equations describing (1) the diffusive and reactive rates of change in concentration of all the molecules in the flame and (2) the flow of the reacting gases. Although the details of hydrocarbon-flame models are still disputed, many of their general features are clear and the basic reactions well-known; computer models ...
The study of chemoattractant receptors on human monocytes had been limited by the lack of a radioligand suitable for use with the small numbers of cells routinely available from human donors. A new synthetic oligopeptide radioligand f(/sup 35/S)Met-leu-phe, with a higher specific radioactivity than was available with the tritiated compound, was used to characterize a chemoattractant receptor on freshly isolated human blood monocytes. These cells bind f(/sup 35/)met-leu-phe with a dissociation constant (KD) of 30.2 +/- 5.6 nM and contain 84,000 +/- 11,300 receptors per cell. f(/sup 35/)met-leu-phe does not bind specifically to blood lymphocytes. The specificity of the oligopeptide receptor on monocytes is indistinguishable from the oligopeptide chemoattractant receptor on human polymorphonuclear leukocytes. Using f(/sup 35/)met-leu-phe, it will now be feasible to study the chemotactic peptide receptor on small numbers of partially purified peripheral blood monocytes ...
Abstract Plant annexins represent a multigene family involved in cellular elongation and development. A cDNA encoding a novel annexin was isolated from a cotton (Gossypium hirsutum) fiber cDNA library and designated-GhAnx1. This gene encodes a 316 amino acid protein with a theoretical molecular mass of 36.06 kDa and a theoretical pI of 6.19. At the amino acid level, it shares high sequence similarity and has evolutionary relationships with annexins from higher plants. The purified recombinant protein expressed in-Escherichia coli-was used to investigate its physicochemical properties. Circular dichroism spectrum analyses showed a positive peak rising to the maximum at 196 nm and a broad negative band rounding 215 nm, suggesting that the GhAnx1 protein was prominently -helical. The fluoresc...
A novel single-chain immunotoxin was constructed by combining a CD33-specific single chain Fv (scFv) antibody fragment with an engineered variant of Pseudomonas exotoxin A (ETA). The variant toxin carries the KDEL peptide at its C-terminus, a cellular peptide mediating improved retrograde transport to the endoplasmic reticulum. The purified recombinant fusion protein induced potent apoptosis of the human myeloid cell lines U937, HL-60 and THP-1. Up to 98% of U937 cells were eliminated after treatment for 72 h with a single dose of 500 ng/ml (c. 7 nmol/l). Killing was antigen-specific and occurred by apoptosis. A control protein, consisting of a CD19-specific scFv antibody fragment fused to the ETA-KDEL toxin, failed to induce death of the CD19-negative cell lines U937, HL-60 and THP-1. The CD33-ETA toxin also mediated apoptosis of fresh patient-derived acute myeloid leukaemia cells from bone marrow and peripheral blood. The pronounced antigen-restricted ...
The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) has two major functions: interacting with the receptor to mediate virus entry and inducing protective immunity. Coincidently, the receptor-binding domain (RBD, residues 318-510) of SAR-CoV S protein is a major antigenic site to induce neutralizing antibodies. Here, we used RBD-Fc, a fusion protein containing the RBD and human IgG1 Fc, as a model in the studies and found that a single amino acid substitution in the RBD (R441A) could abolish the immunogenicity of RBD to induce neutralizing antibodies in immunized mice and rabbits. With a panel of anti-RBD mAbs as probes, we observed that R441A substitution was able to disrupt the majority of neutralizing epitopes in the RBD, suggesting that this residue is critical for the antigenic structure responsible for inducing protective immune responses. We also demonstrated that the RBD-Fc bearing R441A mutation could not bind to soluble and cell-associated ...
A {lambda}gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. ...
A #lambda#gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. ...
PCR detection of genetic material of the parasites present in faeces may be an alternative for microscopic and serological tests routinely used for diagnosing parasitic enteral infections. However, small amount of target DNA combined with low efficiency of total DNA extraction, and presence of PCR inhibitors in the samples to be amplified, may cause false negative detection results. The aim of this work was to evaluate the impact of DNA isolation procedure used on the amplification of DNA fragments from the genomes of protozoan Cryptosporidium parvum and the nematode Trichinella spiralis. Two methods based on different principles of biological material lysis were evaluated; NucliSENS miniMAG employing simultaneously applied chemical lysis and mechanical disruption or mechanical disruption followed by enzymatic lysis in case of QIAamp DNA Stool Mini Kit. Both of the analyzed systems for nucleic acids purification allowed isolation of DNA from purified ...
["3H]Indole-3-acetyl-myo-inositol was hydrolyzed by buffered extracts of acetone powders prepared from 4 day shoots of dark grown Zea mays L. seedlings. The hydrolytic activity was proportional to the amount of extract added and was linear for up to 6 hours at 37"0C. Boiled or alcohol denatured extracts were inactive. Analysis of reaction mixtures by high performance liquid chromatography demonstrated that not all isomers of indole-3-acetyl-myo-inositol were hydrolyzed at the same rate. Buffered extracts of acetone powders were prepared from coleoptiles and mesocotyls. The rates of hydrolysis observed with coleoptile extracts were greater than those observed with mesocotyl extracts. Active extracts also catalyzed the hydrolysis of esterase substrates such as #alpha#-naphthyl acetate and the methyl esters of indoleacetic acid and naphthyleneacetic acid. Attempts to purify the indole-3-acetyl-myo-inositol hydrolyzing activity by chromatographic procedures resulted in ...
Chorismate mutase, a branch-point enzyme in the aromatic amino acid pathway of Saccharomyces cerevisiae, and also a mutant chorismate mutase with a single amino acid substitution in the C-terminal part of the protein have been purified approximately 20-fold and 64-fold from overproducing strains, respectively. The wild-type enzyme is activated by tryptophan and subject to feedback inhibition by tyrosine, whereas the mutant enzyme does not respond to activation by tryptophan nor inhibition by tyrosine. Both enzymes are dimers consisting of two identical subunits of M_r 30,000, each one capable of binding one substrate and one activator molecule. Each subunit of the wild-type enzyme also binds one inhibitor molecule, whereas the mutant enzyme lost this ability. The enzyme reaction was observed by "1H NMR and shows a direct and irreversible conversion of chorismate to prephenate without the accumulation of any enzyme-free intermediates. The kinetic data of the ...
The entrapment of proteins using the sol-gel route provides a means to retain its native properties and artificially reproduce the molecular crowding and confinement experienced by proteins in the cell allowing investigation of the physico-chemical and structural properties of biomolecules at the biotic/abiotic interface. The biomolecules are spatially separated and 'caged' in the gel structure but solutes can freely permeate the matrix. Thus, properties such as the folding of ensembles of individual molecules can be examined in the absence of aggregation effects that can occur in solution studies. Green fluorescent protein from Aequorea coerulescens was used as a model protein to examine the unfolding/re-folding properties of protein in silica gels. The recombinant protein was isolated and purified from Escherichia coli extracts by cell lysis, three-phase partitioning, dialysis, and anion exchange chromatography. The purity of the protein was ...
Vulnerability of 25-hydroxy-(26,27-{sup 3}H)vitamin D{sub 3} 3{beta}-N-(4-azido-2-nitrophenyl)glycinate, a photoaffinity analogue of 25-hydroxyvitamin D{sub 3} (25-OH-D{sub 3}) toward standard conditions of carboxymethylationin promoted the authors to synthesize 25-hydroxyvitamin D{sub 3} 3{beta}-3{prime}-(N-(4-azido-2-nitrophenyl)amino)propyl ether (25-ANE), a hydrolytically stable photoaffinity analogue of 25-OH-D{sub 3}, and 25-hydroxyvitamin D{sub 3} 3{beta}-3{prime}-(N-(4-azido-2-nitro-(3,5-{sup 3}H)phenyl)amino)propyl ether ({sup 3}H-25-ANE), the radiolabeled counterpart of 25-ANE competes for the 25-OH-D{sub 3} binding site in rat serum vitamin D binding protein (rDBP). On the other hand, UV exposure of a sample of purified rat DBP (rDBP), preincubated in the dark with {sup 3}H-25-ANE, covalently labeled the protein. However, very little covalent labeling was observed in the absence of UV light or in the presence of a large excess of 25-OH-D{sub 3}. These ...
(methine-{sup 3}H)1,1-Di-(4-chlorophenyl)-2,2,2-trichloroethane ((methine-{sup 3}H)DDT) and its di-(4-nitrophenyl) analog, both of high purity with a moderately high specific activity were prepared. Chloro-benzene was condensed with (1-{sup 3}H)1-(4-chlorophenyl)-2,2,2-trichloro-ethanol, which has been synthesized by sodium boro({sup 3}H)hydride reduction of 4-chlorophenyl trichloromethyl ketone. The purified ({sup 3}H)DDT had a specific activity of 0.77 Ci/mmol (28.49 GBq/mmol). (methine-{sup 3}H)1,1-Diphenyl-2,2,2-trichloroethane was similarly synthesized and was nitrated to give (methine-{sup 3}H)1,1-di-(4-nitrophenyl)-2,2,2-trichloro-ethane of 1.63 Ci/mmol (60.31 GBq/mmol). Dehydrochlorination with housefly enzyme (glutathione-dependent DDT dehydrochlorinase) showed a remarkable isotope effect. For DDT, the observed tritium isotope effect on V{sub max}/K{sub m} was 11.51{plus minus}0.52. For the nitro-analog, the value was 11.3{plus minus}1.2. We measured ...
Barbiturates substituted at the 5-position with organyltelluroalkyl or organylselenoalkyl groups were prepared by ring annulation of appropriately substituted diethyl malonates with urea or thiourea. The substituted diethyl malonates (phenyltellurobutyl(ethyl), i-propyltelluropropyl(ethyl), i-propyltelluropropyl(allyl), i-propyselenopropyl(ethyl), and phenylselenohexyl(methyl)) were prepared in 55-91% yield by reaction of diethyl omega-bromoalkyl(alkyl)malonates with organyltellurolates or selenolates (RXNa; X = Se, Te) in ethanol/benzene. The following barbiturates were obtained in 28-84% yield: phenyltellurobutyl(ethyl), m.p. 100/sup 0/C; i-propyltelluropropyl(ethyl), m.p. 119/sup 0/C; i-propylselenopropyl(ethyl), m.p. 137/sup 0/C; phenyl-selenohexyl(methyl), m.p. 124/sup 0/C. The 5-substituted thiobarbiturates i-propyltelluropropyl(ethyl)-(m.p. 75/sup 0/C) and i-propylselenopropyl(ethyl)thiobarbiturate (m.p. 83/sup 0/C) were isolated in 32 and 20% yield, respectively. The ...
Full text: HMPAO, a butane monoxime derivative of propylene amine and ECD, ethyl cysteinate dimer, are currently being used as brain imaging agents. In an effort to develop new radiopharmaceuticals for imaging brain, we have synthesized amine and imine derivatives of ethyl cysteinate with butane monoxime, optimized labelling procedure with "9"9"mTc and studied the in vivo behaviour of their "9"9"mTc complexes. Imine derivative of ethyl cysteinate with butane monoxime was synthesized by refluxing equimolar amounts of butane monoxime and ethyl cystinate in dry benzene for 3 hours. The product was purified over silica with 10% ethyl acetate in chloroform as eluent. Amine derivative was prepared by reducing the imine derivative with sodium borohydride. The ligands were characterized by NMR spectroscopy and were found to be pure. The ligands were labelled with "9"9"mTc using stannous chloride as reducing agent. The complexes were characterized by paper chromatography, ...
The central benzodiazepine receptor tracer [N-methyl-"1"1C]iomazenil (Ro 16-0154) was synthesized by alkylation of the desmethyl precursor noriomazenil with ["1"1C]methyl iodide. The ["1"1C]CH_3I (prepared by reduction of ["1"1C]CO_2 with LiAlH_4 followed by reaction with HI) was reacted with noriomazenil inN,N -dimethylformamide and Bu_4N"+OH"- for 1 min at 80 deg. C and purified by HPLC (C_1_8, 34% CH_3CN/H_2O, 7 mL/min). The product was obtained with synthesis time 35 #+-# 5 min (mean #+-# SD, n = 7), radiochemical yield (EOB) 36 #+-# 16%, radiochemical purity 99 #+-# 1%, and specific activity 5100 #+-# 2800 mCi/#mu#mol. Absorbed radiation doses were calculated from previously acquired human biodistribution data. The urinary bladder wall received the highest dose (0.099 mGy/MBq) for 4.8 h voiding interval and the effective dose equivalent was 0.015 mSv/MBq. After i.v. injection of ["1"1C]iomazenil in an adult baboon or healthy human volunteer, radioactivity ...
Eight phospholipases A2 (PLAs) and four three-finger-toxins (3FTx) from the pooled venom of Bungarus fasciatus (Bf) were previously studied and sequenced, but their expression pattern in individual Bf venom and possible geographic variations remained to be investigated. We herein analyze the individual venom of two Bf specimens from Kolkata (designated as KBf) to address this question. Seven PLAs and five 3FTx were purified from the KBf venoms, and respective cDNAs were cloned from venom glands of one of the snakes. Comparison of their mass and N-terminal sequence revealed that all the PLAs were conserved in both KBf venoms, but that two of their 3FTx isoforms were variable. When comparing the sequences of these KBf-PLAs with those published, only one was found to be identical to that of Bf Vb-2, and the other five were 94-98% identical to those of Bf II, III, Va, VI and XI-2, respectively. Notably, the most abundant PLA isoforms of Bf and KBf venoms contain Pro31 ...
A new method that allows rapid preconcentration and separation of plutonium and neptunium in water samples was developed for the measurement of "2"3"7Np and Pu isotopes by inductively-coupled plasma mass spectrometry (ICP-MS) and alpha spectrometry. "2"3"8U can interfere with "2"3"9Pu measurement by ICP-MS as "2"3"8UH"+ mass overlap and "2"3"7Np via peak tailing. The method provide enhanced removal of uranium by separating Pu and Np initially on TEVA Resin, then moving Pu to DGA resin for additional removal of uranium. The decontamination factor for uranium from Pu is almost 100,000 and the decontamination factor for U from Np is greater than 10,000. This method uses stacked extraction chromatography cartridges and vacuum box technology to facilitate rapid separations. Preconcentration is performed using a streamlined calcium phosphate precipitation method. Purified solutions are split between ICP-MS and alpha spectrometry so that long and short-lived Pu isotopes ...
3-Quinuclidinyl benzilate (QNB) is a potent muscarinic antagonist that binds to muscarinic acetylcholine receptors. Iodination of QNB is difficult because it contains rings that are deactivated, while phenol rings are activated. The purpose of this research is to develop an appropriate method for the iodination of QNB at no-carrier-added concentrations of /sup 125/I/NaI. Phenol was chose as the test compound for direct iodination by adding chloramine T and nonradioactive sodium iodide to the phenol in a phosphate buffer and methanol system. A simple method to radioiodinate QNB at several concentrations was developed in situ in the presence of QNB, thallic trifluoroacetate /sup 125/I/NaI, /sup 127/I/NaI, and aluminium chloride. Using milligram amounts of QNB (7.12 x 10/sup -6/ moles), three reaction steps were involved: QNB was reacted with thallic trifluoroacetate at 60/sup 0/C for 24 hours, then radioactive /sup 125/I/NaI and aluminium chloride were added to the reaction mixture and ...
The thermal neutron capture cross sections and the neutron capture resonance integrals of {sup 241}Am leading to the production of the isomer {sup 242m}Am and the ground-state {sup 242g}Am were measured radiochemically by the Cd-ratio technique with neutron flux monitors of Co/Al and Au/Al alloy. Highly-purified {sup 241}Am targets were irradiated in an aluminum capsule by using JMTR. The neutron fluxes and their epithermal neutron fractions were determined by measuring {gamma}-rays of {sup 60}Co and {sup 198}Au. The yields of {sup 242m}Am and {sup 242g}Am were decided by analyzing growth and decay curves of the {alpha}-ray activity ratios {sup 242}Cm/{sup 241}Am. The resultant thermal neutron capture cross sections are 85.7 {+-} 6.3 b and 768 {+-} 58 b for {sup 242m}Am and {sup 242g}Am, and the resonance integrals 114 {+-} 7 b and 1,694 {+-} 146 b, respectively. The differences between the present results and the evaluated values by Mughabghab are 38-59%. The ...
A new method that allows rapid preconcentration and separation of plutonium and neptunium in water samples was developed for the measurement of {sup 237}Np and Pu isotopes by inductively-coupled plasma mass spectrometry (ICP-MS) and alpha spectrometry; a hybrid approach. {sup 238}U can interfere with {sup 239}Pu measurement by ICP-MS as {sup 238}UH{sup +} mass overlap and {sup 237}Np via peak tailing. The method provide enhanced removal of uranium by separating Pu and Np initially on TEVA Resin, then moving Pu to DGA resin for additional removal of uranium. The decontamination factor for uranium from Pu is almost 100,000 and the decontamination factor for U from Np is greater than 10,000. This method uses stacked extraction chromatography cartridges and vacuum box technology to facilitate rapid separations. Preconcentration is performed using a streamlined calcium phosphate precipitation method. Purified solutions are split between ICP-MS and alpha spectrometry so ...
A cryogenic separation method of chloromethane (CH{sub 3}Cl) from methyl iodide (CH{sub 3}I) for {delta}{sup 37}Cl measurement with isotope ratio mass spectrometer is described. A cold trap with sealed 2,2,4-trimethylpentane (TMP) as the cryogen is used in this method. CH{sub 3}Cl can be separated from CH{sub 3}I at the TMP melting point (-107 deg. C) based on the difference in the vapor pressure between CH{sub 3}Cl (322.6 Pa) and CH{sub 3}I (lower than 1.3 Pa) at -107 deg. C. After two-step separation processes, the yields of CH{sub 3}Cl purified from CH{sub 3}Cl-CH{sub 3}I mixture are 96-101%, and the difference between the {delta}{sup 37}Cl{sub original} and {delta}{sup 37}Cl{sub Afterseparation} is from -0.06 to +0.06 per mille (0.01 {+-} 0.04 per mille ). These results suggest that CH{sub 3}Cl is completely separated from CH{sub 3}I with no change of {delta}{sup 37}Cl value. This method using the cold trap with sealed TMP is very safe and not harmful, because ...
(-)-N6-(R-4-Hydroxyphenylisopropyl)adenosine (HPIA) was iodinated with NaI and trace /sup 125/I. Mono- and diiodinated reaction products and the starting material were separated by high pressure liquid chromatography and the structures of the reaction products were verified by NMR. (-)-N6-(R-Phenylisopropyl)adenosine (PIA), IHPIA, and I2HPIA decreased rat atrial contractility with ED50 values of 24, 28, and 33 nM, respectively. The contractile effects of these compounds were competitively blocked by theophylline (KI . 7.9 microM), but were not affected by adenosine deaminase. IHPIA also inhibited (-)isoproterenol-stimulated cyclic AMP accumulation in adipocytes with an ED50 (10 nM) and to an extent (83%) nearly identical to PIA. (/sup 125/I)HPIA prepared using carrier-free /sup 125/I bound to adenosine receptors on membranes from rat cerebral cortex, adipocyte ghosts, and heart ventricles. Binding was inhibited stereospecifically by PIA and by other adenosine analogues and ...
Four trypsin inhibitor homologs, the first known from Dendroaspis angusticeps venom, were characterized using a combination of gel filtration, cation exchange, reverse-phase liquid chromatography, Edman degradation and mass spectrometry. The four toxins comprise two 57 residue and two 59 residue isoforms. The long toxins possess a Lys-Gln N-terminal extension lacked by the short toxins. The only other structural difference is an Arg/His replacement at position 55. The long Arg55 variant is identical to trypsin inhibitor E from the venom of Dendroaspis polylepis. The name epsilon-dendrotoxin is suggested so as to follow the nomenclature of Benishin, C.G., Sorensen, R.G., Brown, W.E., Krueger, B.K., Blaustein, M.P., 1988. Four polypeptide components of green mamba venom selectively block certain potassium channels in rat brain synaptosomes. Mol. Pharmacol. 34, 152-159. Among snake venom protease inhibitors, the epsilon-dendrotoxins are structurally most like the delta-dendrotoxins, with ...
Research at the interface between biomolecules and inorganic nanocrystals has resulted in a great number of new discoveries. In part this arises from the synergistic duality of the system: biomolecules may act as self-assembly agents for organizing inorganic nanocrystals into functional materials; alternatively, nanocrystals may act as microscopic or spectroscopic labels for elucidating the behavior of complex biomolecular systems. However, success in either of these functions relies heavily uponthe ability to control the conjugation and assembly processes.In the work presented here, we first design a branched DNA scaffold which allows hybridization of DNA-nanocrystal monoconjugates to form discrete assemblies. Importantly, the asymmetry of the branched scaffold allows the formation of asymmetric2assemblies of nanocrystals. In the context of a self-assembled device, this can be considered a step toward the ability to engineer functionally distinct inputs and outputs.Next we develop an ...
ADP-ribosylation factors (ARFs) are small guanine nucleotide-binding proteins that enhance the enzymatic activities of cholera toxin. Two ARF cDNAs, ARF1 and ARF3, were cloned from a human cerebellum library. Based on deduced amino acid sequences and patterns of hybridization of cDNA and oligonucleotide probes with mammalian brain poly(A)"+ RNA, human ARF1 is the homologue of bovine ARF1. Human ARF3, which differs from bovine ARF1 and bovine ARF2, appears to represent a newly identified third type of ARF. Hybridization patterns of human ARF cDNA and clone-specific oligonucleotides with poly(A)"+ RNA are consistent with the presence of at least two, and perhaps four, separate ARF messages in human brain. In vitro translation of ARF1, ARF2, and ARF3 produced proteins that behaved, by SDS/PAGE, similar to a purified soluble brain ARF. Deduced amino acid sequences of human ARF1 and ARF3 contain regions, similar to those in other G proteins, that are believed to be ...
A cDNA library was constructed using RNA isolated from the livers of chickens which had been treated with zinc. This library was screened with a RNA probe complementary to mouse metallothionein-I (MT), and eight chicken MT cDNA clones were obtained. All of the cDNA clones contained nucleotide sequences homologous to regions of the longest (375 bp) cDNA clone. The latter contained an open reading frame of 189 bp, and the deduced amino acid sequence indicates a protein of 63 amino acids of which 20 are cysteine residues. Amino acid composition and partial amino acid sequence analyses of purified chicken MT protein agreed with the amino acid composition and sequence deduced from the cloned cDNA. Amino acid sequence comparison establish that chicken MT shares extensive homology with mammalian MTs. Southern blot analysis of chicken DNA indicates that the chicken MT gene is not a part of a large family of related sequences, but rather is likely to be a unique gene ...
High molecular weight DNA from pleroceroid larvae of the tapeworm Spirometra mansonoides was purified from isolated nuclei by conventional techniques. The DNA so isolated has a melting temperature (Tm) of 87 degrees C and a guanine plus cytosine (G/C) content of 44%. 5-Methyl cytosine could not be detected in plerocercoid DNA by HPLC analysis of DNA hydrolysates, by radiolabeling 5'-termini of MspI digests with polynucleotide kinase, or by comparing restriction patterns generated by MspI and HpaII. Renaturation kinetics demonstrated that the genome of S. mansonoides contains repetitive as well as single copy sequences and has a genome size estimated at approx. 1.6 X 10(9) bp. Hybridization was carried out between plerocercoid DNA and cDNAs for human beta-actin, alpha-tubulin and growth hormone (hGH). Rationale for this analysis was based on known homologies among actin and tubulin genes in numerous species and on apparent similarities between hGH and a plerocercoid ...
Immidacloprid is a cyclodiene organochlorine used as an insecticide all over the world and possessing a serous environmental threat. It is mostly used for cotton insects (bollworm, aphid and white fly). For isolation of imidacloprid degrading bacteria, two soil samples were collected from industrial contaminated sites of Kala Shah Kahu district sheikupura, having ten year history of use. Soil samples were analyzed by measuring pH and electric conductivity. The isolation of imidacroprid degrading bacteria was performed by enrichment technique. Eight bacterial strains, S/sub 1-a/ S/2-2-b/ S/2-c/ S/2-d/ S/2-e/ S/sub 2-f/ and S/sub 2-g/ and S/sub e-a/ were isolated on the basis of their colony morphologies. The purified colonies were characterized morphologically, physiologically and biochemically. Gram staining was done and Gram negative strain were confirmed on MacConkey agar and Eosin Methylene Blue. Bacterial strains were also checked for different minimal media in ...
The purified integral membrane protein, band 3, from human erythrocytes was inserted into egg lecithin liposomes. The insertion of band 3 was determined from thermal transition data from the analysis of the C-H stretching region bands recorded at temperatures from 25 to -22[sup o]C. Raman spectra show that band 3 considerably broadens and lowers the thermal transition of egg lecithin liposomes, suggesting the insertion of band 3. The band 3-inserted liposomes were irradiated with gamma-rays (40 Gy) and the radiation target groups were determined by the analysis of the structural sensitive Raman bands in the 1600-1700 cm[sup -1] (amide I), 1200-1300 cm[sup -1] (amide III) and 550-1030 cm[sup -1] (side chain amino groups) regions. The radiation-sensitive groups as identified from Raman spectra in the region 550-1030 cm[sup -1] are tyrosines and cysteines. The radiation-induced changes in the secondary structure were determined from amide I and III bands. Quantitative ...
The purified integral membrane protein, band 3, from human erythrocytes was inserted into egg lecithin liposomes. The insertion of band 3 was determined from thermal transition data from the analysis of the C-H stretching region bands recorded at temperatures from 25 to -22"oC. Raman spectra show that band 3 considerably broadens and lowers the thermal transition of egg lecithin liposomes, suggesting the insertion of band 3. The band 3-inserted liposomes were irradiated with gamma-rays (40 Gy) and the radiation target groups were determined by the analysis of the structural sensitive Raman bands in the 1600-1700 cm"-"1 (amide I), 1200-1300 cm"-"1 (amide III) and 550-1030 cm"-"1 (side chain amino groups) regions. The radiation-sensitive groups as identified from Raman spectra in the region 550-1030 cm"-"1 are tyrosines and cysteines. The radiation-induced changes in the secondary structure were determined from amide I and III bands. Quantitative estimation using the ...
From the alkalised crude extract of Symphytum cordatum (L.) W.K. roots, pyrrolizidine alkaloids (PAs) were extracted as free tertiary bases and polar N-oxides in a merely one-step liquid-liquid partitioning (LLP) in separation funnel and subsequently pre-fractionated by preparative multiple-development (MD) thin-layer chromatography (TLC) on silica gel plates. In this way three alkaloid fractions of different polarities and retention on silica gel plates were obtained as: the most polar N-oxides of the highest retention, the tertiary bases of medium retention, and diesterified N-oxides of the lowest retention. The former fraction was reduced into free bases by sodium hydrosulfite and purified by LLP on Extrelut-NT3 cartridge. It was further analysed together with the two other fractions by high-performance liquid chromatography (HPLC)-ion-trap mass spectrometry with atmospheric pressure chemical ionization (APCI) interface on XTerra C{sub 18} column using a ...
Incorporation of /sup 32/P from (gamma-32P)ATP into tyrosine residues of the insulin-like growth factor (IGF)-II receptor was observed in a Triton X-100-insoluble fraction of rat adipocyte plasma membranes. IGF-II receptor phosphorylation proceeded to a stoichiometry of approximately 0.5 mol of phosphate/IGF-II binding site after 10 min of incubation at 4 degrees C. A Km for ATP of 6 microM was calculated for this phosphorylation reaction. Addition of IGF-II caused an approximately 2-fold increase in tyrosine phosphorylation of the IGF-II receptor in this preparation. In contrast, phosphorylation of angiotensin II by the Triton X-100 washed membranes was not stimulated by IGF-II. Incubation of purified receptor immobilized on IGF-II agarose or of receptor-enriched low density microsomal membranes with (gamma-32P)ATP did not result in appreciable incorporation of (/sup 32/P)phosphate into the IGF-II receptor nor into exogenous substrates. These data suggest that the ...
A major regulatory element required for expression of the human [alpha]-globin genes is located 40 kb upstream of the embryonic [zeta]-globin gene. To understand how this and other locus control region (LCR) elements contribute to high-level expression in erythroid cells, we have performed high-resolution, in vivo dimethyl sulfate footprinting. In addition, we have modified the dimethyl sulfate-based ligation-mediated polymerase chain reaction in vivo footprinting procedure to permit the assessment of interactions at guanine and adenine residues, rather than guanines alone. In vivo footprinting of the human [alpha]-LCR element carried on chromosome 16 in a mouse erythroleukemia cell environment revealed protein occupancy at GATA-1, AP-1/NF-E2, and CACC/GGTGG motifs, specific differences compared with in vitro protein binding, and distinct changes in one region upon dimethyl sulfoxide-induced cellular maturation. No protein contacts were detected in nonexpressing hepatoma cells. In ...
Genes encoding mutants of the thymine photodimer repair enzyme from bacteriophage T4 (T4 endonuclease V) having an amino acid substitution (T127M, W128A, W128S, Y129A, K130L, Y131A, Y132A) were constructed by use of a previously obtained synthetic gene and expressed in Escherichia coli under the control of the E. coli tryptophan promoter. An in vitro assay of partially fractionated mutant proteins for glycosylase activity was performed with chemically synthesized substrates containing a thymine photodimer. T127M and K130L showed almost the same activity as the wild-type protein. Although W128S, Y131A, and Y132A were slightly active, W128A and Y129A lost activity. The results indicated that the aromatic amino acids around position 130 may be important for the glycosylase activity. Mutant T127M was purified, and the Km value was found to be of the same order as that of the wild type (10(-8) M). In vivo activities for all mutants were characterized with UV-sensitive ...
Partially purified UDPgalactosyltransferase (EC 2.4.1.22) from bovine milk has been used to synthesize millimolar amounts of compounds such as Gal..beta..(1..-->..4)Glc, Gal..beta..(1..-->..4)GlcNAc-..beta..-hexanolamine, and Gal..beta..(1..-->..4)-GlcNAc..beta..(1..-->..4)GlcNAc. The same method has been used to prepare similar compounds containing /sup 13/C-enriched galactopyranosyl moieties. Gal..beta..(1..-->..4)GlcNAc-..beta..-hexanolamine was also synthesized in a solid-phase system in which the GlcNAc-..beta..-hexanolamine glycoside was covalently linked to agarose beads. At pH 7.0 and at 1 to 5 mM Mn/sup 2 + +/ the yields of the galactosyl saccharides are greater than 90% by using 10% excess of UDPGal donor. The use of a 90% enriched (1-/sup 13/C)galactosyl residue allowed the determination of the most abundant conformer about the galactopyranosyl-glycoside linkage by analysis of the carbon-carbon coupling constants from Cl to Gal to the ...
The authors have investigated insulin receptor phosphorylation in differentiated cultured BC3H-1 myocytes. As for other insulin-responsive cell systems in partially purified wheat germ agglutinin receptor preparations, insulin stimulates the phosphorylation of its own receptor (95K ..beta..-subunits) in a dose dependent manner (0-400 nM), as identified by immunoprecipitation with antiinsulin receptor antibodies and SDS-PAGE. In the same preparations they show that 12-0-tetradecanyl phorbol acetate (TPA), which in many respect ..beta..-subunits in the same dose dependent manner (0-5 ..mu..M). In addition, antiinsulin receptor antibodies (B-10) also induced phosphorylation of mimics insulin action, also induced phosphorylation of the insulin receptor and HPLC tryptic maps of the /sup 32/P-labeled ..beta..-subunit were identical to those for insulin-induced receptor phosphorylation. However, while insulin and TPA are potent stimulators of glucose transport in these ...
Autoreactive cytotoxic CD8 T-cells (CTLs) play a key pathogenic role in the destruction of insulin-producing beta-cells resulting in type 1 diabetes. However, knowledge regarding their targets is limited, restricting the ability to monitor the course of the disease and immune interventions. In a multi-step discovery process to identify novel CTL epitopes in human preproinsulin (PPI), PPI was digested with purified human proteasomes, and resulting COOH-fragments aligned with algorithm-predicted HLA-binding peptides to yield nine potential HLA-A1, -A2, -A3 or -B7-restricted candidates. An UV-exchange method allowed the generation of a repertoire of multimers including low-affinity HLA-binding peptides. These were labeled with quantum dot-fluorochromes and encoded in a combinatorial fashion, allowing parallel and sensitive detection of specific, low-avidity T-cells. Significantly increased frequencies of T-cells against four novel PPI epitopes (PPI(4-13)/B7, ...
The active site of chicken gizzard myosin was labeled by direct photoaffinity labeling with ["3H]UDP. ["3H] UDP was stably trapped at the active site by addition of vanadate (Vi) and Co"2"+. The extraordinary stability of the myosin.Co2+.[3H]UDP.Vi complex (t1/2 greater than 5 days at 0 degrees C) allowed it to be purified free of extraneous ["3H]UDP before irradiation began. Upon UV irradiation, greater than 60% of the trapped ["3H]UDP was photoincorporated into the active site. Only the 200-kDa heavy chain was labeled, confirming earlier results using ["3H]UTP. Extensive tryptic digestion of photolabeled myosin subfragment 1 followed by high performance liquid chromatography separations and removal of nucleotide phosphates by treatment with alkaline phosphatase allowed two labeled peptides to be isolated. Sequencing of the labeled peptides and radioactive counting showed that Glu185 was the residue labeled. Since UDP is a zero-length cross-linker, Glu185 is ...
It was the aim of this study to specifically detect the DNA sequences for the bphC gene, the meta-cleavage enzyme of the aerobic catabolic pathway for biphenyl and polychlorinated biphenyl degradation, in aquatic sediments without prior cultivation of microorganisms by using extraction of total DNA, PCR amplification of bphC sequences, and detection with specific gene probes. The direct DNA extraction protocol used was modified to enhance lysis efficiency. Crude extracts of DNA were further purified by gel filtration, which yielded DNA that could be used for the PCR. PCR primers were designed for conserved regions of the bphC gene from a sequence alignment of five known sequences. The specificity of PCR amplification was verified by using digoxigenin-labeled DNA probes which were located internal to the amplified gene sequence. The detection limit for the bphC gene of Pseudomonas paucimobilis Q1 and Pseudomaonas sp. strain LB 400-like sequences for the bphC gene ...
Rapid detection and characterization of food borne pathogens such as Escherichia coli O157:H7 is crucial for epidemiological investigations and food safety surveillance. As an alternative to conventional technologies, we examined the sensitivity and specificity of nucleic acid microarrays for detecting and genotyping E. coli O157:H7. The array was composed of oligonucleotide probes (25-30 mer) complementary to four virulence loci (intimin, Shiga-like toxins I and II, and hemolysin A). Target DNA was amplified from whole cells or from purified DNA via single or multiplexed polymerase chain reaction (PCR), and PCR products were hybridized to the array without further modification or purification. The array was 32-fold more sensitive than gel electrophoresis and capable of detecting amplification products from < 1 cell equivalent of genomic DNA (1 fg). Immunomagnetic capture, PCR and a microarray were subsequently used to detect 55 CFU ml-1 (E. coli O157:H7) ...
In order to optimize the performance of molecular organic electronic devices it is important to study the intermolecular density of states and charge transport mechanisms in the environment of crystalline organic material. Using this approach in Field Effect Transistors (FETs) we show that material purification improves carrier mobility and decreases density of the deep localized electronic state. We also report a general exponential energy dependence of the density of localized states in a vicinity of the mobility edge (Fermi energies up to approx7 times higher than the thermal energy (kT)) in a variety of the extensively purified molecular organic crystal FETs. This observation and the low activation energy of the order of approxkT suggest that molecular structural misplacements of the sizes that are comparable with thermal molecular modes rather than impurity deep traps play a role in formation of these shallow states. We find that the charge carrier mobility in ...
A unique feature of cyanobacteria genomes is the abundance of genes that code for hypothetical proteins containing tandem pentapeptide repeats approximately described by the consensus motif A(N/D)LXX. To date, the structures of two pentapeptide-repeat proteins (PRPs) have been determined, with the tandem pentapeptide-repeat sequences observed to adopt a novel type of right-handed quadrilateral ?-helix, or Rfr-fold, in both structures. One structure, Mycobacterium tuberculosis MfpA, is a 183-residue protein that contains 30 consecutive pentapeptide repeats and appears to offer antibiotic resistance by acting as a DNA mimic. The other structure, Cyanothece 51142 Rfr32, is a 167-residue protein that contains 21 consecutive pentapeptide repeats. The function of Rfr32, like the other 35 hypothetical PRPs identified in the genome of Cyanothece, is unknown. In an effort to understand the role of PRPs in cyanobacteria and to better characterize the structural properties of Rfr-folds with ...
Duchenne and Becker muscular dystrophies are caused by defects of dystrophin, which forms a part of the membrane cytoskeleton of specialized cells such as muscle. It has been previously shown that the dystrophin-associated protein A1 (59-kDa DAP) is actually a heterogeneous group of phosphorylated proteins consisting of an acidic ({alpha}-A1) and a distinct basic ({beta}-A1) component. Partial peptide sequence of the A1 complex purified from rabbit muscle permitted the design of oligonucleotide probes that were used to isolate a cDNA for one human isoform of A1. This cDNA encodes a basic A1 isoform that is distinct from the recently described syntrophins in Torpedo and mouse and is expressed in many tissues with at least five distinct mRNA species of 5.9, 4.8, 4.3, 3.1, and 1.5 kb. A comparison of the human cDNA sequence with the GenBank expressed sequence tag (EST) data base has identified a relative from human skeletal muscle, EST25263, which is probably a human ...
A carrier-free radioiodinated phenylazide derivative of forskolin, 3-iodo-4-azidophenethylamido-7-O-succinyl-deacetyl-forskolin (({sup 125}I)IAPS-forskolin), has been shown to be a highly selective photoaffinity probe for the human erythrocyte glucose transported and the glucose transport proteins found in several mammalian tissues and cultured cells where the glucose transport protein is present at a low concentration. The photoincorporation of ({sup 125}I)IAPS-forskolin into these glucose transporters was blocked by D- (but not L-) glucose, cytochalasin B, and forskolin. In addition to labeling the mammalian glucose transport proteins, ({sup 125}I)IAPS-forskolin also labeled the L-arabinose transporter from E. coli. In muscle and adipose tissues, glucose transport is markedly increased in response to insulin. ({sup 125}I)IAPS-forskolin was shown to selectivity tag the glucose transporter in membranes derived from these cells. In addition, the covalent derivatization of the transport ...
A carrier-free radioiodinated phenylazide derivative of forskolin, 3-iodo-4-azidophenethylamido-7-O-succinyl-deacetyl-forskolin (["1"2"5I]IAPS-forskolin), has been shown to be a highly selective photoaffinity probe for the human erythrocyte glucose transported and the glucose transport proteins found in several mammalian tissues and cultured cells where the glucose transport protein is present at a low concentration. The photoincorporation of ["1"2"5I]IAPS-forskolin into these glucose transporters was blocked by D- (but not L-) glucose, cytochalasin B, and forskolin. In addition to labeling the mammalian glucose transport proteins, ["1"2"5I]IAPS-forskolin also labeled the L-arabinose transporter from E. coli. In muscle and adipose tissues, glucose transport is markedly increased in response to insulin. ["1"2"5I]IAPS-forskolin was shown to selectivity tag the glucose transporter in membranes derived from these cells. In addition, the covalent derivatization of the transport protein in ...
We examined a catalytic effect of niobium oxide (Nb{sub 2}O{sub 5}) on the hydrogen storage properties of MgH{sub 2} prepared by mechanical ball milling method. The MgH{sub 2} composite doped with 1 mol% Nb{sub 2}O{sub 5} by ball milling for 20 h desorbed hydrogen up to {approx}6 mass% in the temperature range from 200 to 250 {sup o}C at the heating rate of 5 {sup o}C/min under a purified helium flow. After dehydrogenation at 200 {sup o}C, the product showed remarkable hydrogen absorption kinetics. A large amount of gaseous hydrogen up to {approx}4.5 mass% was absorbed even at room temperature under 1 MPa hydrogen pressure within 15 s and finally its capacity reached up to 5 mass%. Furthermore, the valence state of Nb{sub 2}O{sub 5} doped in MgH{sub 2} was examined by X-ray absorption near edge structure (XANES) measurement. The results indicated that additive Nb{sub 2}O{sub 5} was reduced by MgH{sub 2} during mechanical milling. This suggests that the Nb compound, ...
Anti-carcinoembryonic antigen monclonal antibody (MAb) CEA 102 was produced by immunization with purified CEA and the specific accumulation of radiolabeled CEA 102 in colorectal cancers was investigated by autoradiography of sugical specimens using Fuji Computed Radiography (FCR). Five patients with colorectal cancer were injected intravenously with {sup 131}I-labeled intact CEA 102 or its F(ab'){sub 2}. Primary tumor and liver metastases were successfully detected by external scanning with a gamma camera in 4 cases. Autoradiographic study of the surgical specimens using FCR showed predominant localization of {sup 131}I-labeled CEA 102 in primary tumors and liver metastases in all cases. Even a small liver metastasis (0.5 cm) was clearly visualized in the autoradiogram by FCR. The pixel distribution curves of the density of the respective tissues in the autoradiograms by FCR showed the heterogeneity of the distribution of administreted radiolabeled MAb in ...
Anti-carcinoembryonic antigen monclonal antibody (MAb) CEA 102 was produced by immunization with purified CEA and the specific accumulation of radiolabeled CEA 102 in colorectal cancers was investigated by autoradiography of sugical specimens using Fuji Computed Radiography (FCR). Five patients with colorectal cancer were injected intravenously with "1"3"1I-labeled intact CEA 102 or its F(ab')_2. Primary tumor and liver metastases were successfully detected by external scanning with a gamma camera in 4 cases. Autoradiographic study of the surgical specimens using FCR showed predominant localization of "1"3"1I-labeled CEA 102 in primary tumors and liver metastases in all cases. Even a small liver metastasis (0.5 cm) was clearly visualized in the autoradiogram by FCR. The pixel distribution curves of the density of the respective tissues in the autoradiograms by FCR showed the heterogeneity of the distribution of administreted radiolabeled MAb in individual tumors, ...
The three main objectives of this study were: (1) to determine if depletion of proteoglycans from the cartilage matrix that occurs during osteoarthritis causes a measurable increase of cartilage proteoglycan components in the synovial fluid and sera, (2) to observe what effect intracellular cAMP has on the expression of matrix components by chondrocytes, and (3) to determine if freshly isolated chondrocytes contain detectable levels of mRNA for fibronectin. Canine serum keratan sulfate and hyaluronate were measured to determine if there was an elevation of these serum glycosaminoglycans in a canine model of osteoarthritis. A single intra-articular injection of chymopapain into a shoulder joint increased serum keratan sulfate 10 fold and hyaluronate less than 2 fold in 24 hours. Keratan sulfate concentrations in synovial fluids of dogs about one year old were unrelated to the presence of spontaneous cartilage degeneration in the joints. High keratan sulfate in synovial fluids correlated ...
We previously reported the identification of a new family of plant methyltransferases (MTs), named the SABATH family, that use S-adenosyl-l-methionine (SAM) to methylate a carboxyl moiety or a nitrogen-containing functional group on a diverse array of plant compounds. The Arabidopsis genome alone contains 24 distinct SABATH genes. To identify the catalytic specificities of members of this protein family in Arabidopsis, we screened recombinantly expressed and purified enzymes with a large number of potential substrates. Here, we report that the Arabidopsis thaliana gene At3g44860 encodes a protein with high catalytic specificity towards farnesoic acid (FA). Under steady-state conditions, this farnesoic acid carboxyl methyltransferase (FAMT) exhibits K{sub M} values of 41 and 71 {mu}M for FA and SAM, respectively. A three-dimensional model of FAMT constructed based upon similarity to the experimentally determined structure of Clarkia breweri salicylic acid ...
MnO2 in pyrolusite can react with SO2 in flue gas and obtain by-product MnSO4 x H2O. A pilot scale jet bubbling reactor was applied in this work. Different factors affecting both SO2 absorption efficiency and Mn2+ extraction rate have been investigated, these factors include temperature of inlet gas flue, ration of liquid/solid mass flow rate (L/S), pyrolusite grade, and SO2 concentration in the inlet flue gas. In the meantime, the procedure of purification of absorption liquid was also discussed. Experiment results indicated that the increase of temperature from 30 to 70 K caused the increase of SO2 absorption efficiency from 81.4% to 91.2%. And when SO2 concentration in the inlet flue gas increased from 500 to 3000 ppm, SO2 absorption efficiency and Mn2+ extraction rate decreased from 98.1% to 82.2% and from 82.8% to 61.7%, respectively. The content of MnO2 in pyrolusite had a neglectable effect on SO2 absorption efficiency. Low L/S was good for both removal of SO2 and Mn2+ ...
The characteristics of per-fluorocarbon compounds (PFC) are colorless, non-toxic, easily vaporized and nonflammable. Also, some of them are liquids of a high density, low surface tension, low latent heat and low specific heat. These particular chemical and physical properties of fluoro-organic compounds permit their use in very different fields such as electronics, medicine, tribology, nuclear and material science. The Sonatol process was developed under a contract with the DOE. The Sonatol process uses an ultrasonic agitation in a PFC solution that contains a fluorinated surfactant to remove radioactive particles from surfaces. Filtering the suspended particles allows the solutions to be reused indefinitely. They applied the Sonatol process to the decontamination of a heterogeneous legacy Pu-238 waste that exhibited an excessive hydrogen gas generation, which prevents a transportation of such a waste to a Waste Isolation Pilot Plant. Korea Atomic Energy Research Institute (KAERI) is ...
Classical semiconductor physics has been continuously improving electronic components such as diodes, light-emitting diodes, solar cells and transistors based on highly purified inorganic crystals over the past decades. Organic semiconductors, notably polymeric, are a comparatively young field of research, the first light-emitting diode based on conjugated polymers having been demonstrated in 1990. Polymeric semiconductors are of tremendous interest for high-volume, low-cost manufacturing (''printed electronics''). Due to their rather simple device structure mostly comprising only one or two functional layers, polymeric diodes are much more difficult to optimize compared to small-molecular organic devices. Usually, functions such as charge injection and transport are handled by the same material which thus needs to be highly optimized. The present work contributes to expanding the knowledge on the physical mechanisms determining ...
One of the refinements of modern Nuclear Medicine is the capacity of providing dynamic and kinetics images of the administered radiopharmaceutical, reproducing its transport mechanism, action sites, receptor binding and excretion route. With the continues technological advances new radiopharmaceuticals have been developed in order to express higher specificity and with higher characters of affinity between receptor/complex. One radiopharmaceutical is formed by a reagent or bio molecule that has in its structure a radioisotope, that has the objectives of carrying it to the organs of affinity or to benign or malign tumoral process. Somatostatin inhibits the growing and proliferation of several tumoral cells. Somatostatin analogs bind to somatostatic receptors that are expressed in different kind of neoplasia DOTA-LANREOTIDE (DOTALAN) is an octapeptide analog to somatostatin. The interest of labeling the bio conjugate with gallium-67 in Nuclear Medicine comes from its physical, chemical ...
Shewanella oneidensis MR-1 is a model environmental organism that possesses diverse respiratory capacities, including the ability to reduce soluble Cr(VI) to sparingly soluble, less toxic Cr(III). Effective bioremediation of Cr-contaminated sites requires knowledge of the molecular mechanisms and regulation of heavy metal resistance and biotransformation by dissimilatory metal-reducing bacteria. Towards this goal, our ERSP-funded work is focused on the identification and functional analysis of genes/proteins comprising the response pathways for chromate detoxification and/or reduction. Previous transcriptomic profiling and whole-cell proteomic analyses implicated the involvement of a functionally undefined DNA-binding response regulator (SO2426) and a putative azoreductase (SO3585) in the chromate stress response of MR-1. Here we describe a detailed functional analysis of SO2426 and SO3585 in order to begin to understand the role of these proteins in the cellular response to chromate. ...
Enzyme-linked Immunosorbent assay systems for the identification of irradiated egg, pork and chicken was developed. Eggs were irradiated in their shells to 0.5{approx}7kGy. Pork was irradiated to 0.5{approx}3kGy and chicken irradiated to 0.5kGy{approx}5kGy. The most sensitive proteins to irradiation were screened by SDS-PAGE and purified. Ovalbumin from egg, salt soluble protein(p) from pork, and salt soluble protein(c) from chicken showed the most sensitivity to irradiation. To investigate for a practical use in identifying of irradiated egg, pork and chicken, competitive ELISA was performed. The binding activity of ovalbumin to anti-ovalbumin IgG was reduced in a dose-dependent manner by irradiating up to 7kGy, and considerably lowered after irradiating at 7kGy. The concentration of 50% inhibition of ovalbumin to IgG was increased to 1.5(0.5kGy){approx}3.7(7kGy) times in an dose-dependent relationship. The binding activity of salt soluble protein(p) to anti-salt ...
Measurements using radioactive targets are important for the determination of key reaction path ways associated with the synthesis of the elements in nuclear astrophysics (sprocess), advanced fuel cycle initiative (transmutation of radioactive waste), and stockpile stewardship. High precision capture cross-section measurements are needed to interpret observations, predict elemental or isotopical ratios, and unobserved abundances. There are two new detector systems that are presently being commissioned at Los Alamos National Laboratory for very precise measurements of (n,{gamma}) and (n,f) cross-sections using small quantities of radioactive samples. DANCE (Detector for Advanced Neutron-Capture Experiments), a 4 {pi} gamma array made up of 160 BaF{sub 2} detectors, is designed to measure neutron capture cross-sections of unstable nuclei in the low-energy range (thermal to {approx}500 keV). The high granularity and high detection efficiency of DANCE, combined with the high TOF-neutron ...
This thesis deals with the evaluation of Solar Grade Silicon (SoG-Si) purified by different techniques, and also the fabrication and characterization of high efficiency and advanced bifacial solar cells. In the beginning of Chapter 1, various SoG-Si production methods relevant for this work are qualitatively described. The three feedstock materials used in this work are from the Fluidized Bed Reactor (FBR) process, metallurgical feedstock-I and feedstock-II process. In metallurgical feedstock-I, the lifetime of the minority charge carriers in multicrystalline silicon (mc-Si) samples at the grain boundaries are found to be higher than the grains themselves possibly due to lower resistivities in the grain boundaries. The efficiency of the best solar cell obtained using the mc-Si metallurgical feedstock-I is 16.1%. It has been identified that the fast light induced degradation, whose magnitude is lower than that of a reference cell suggests the formation of a B-metal ...