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PubMed2006

DNA and adenoviral vectors encoding carcinoembryonic antigen fused to immunoenhancing sequences augment antigen-specific immune response and confer tumor protection.

A panel of vectors was constructed to encode carcinoembryonic antigen (CEA) fused at its C-terminal end to various polypeptides, so as to compare their immunogenicity by plasmid DNA immunization and adenovirus injection in wild-type and CEA transgenic (CEA.tg) mice. Fusions between CEA and the minimized domain of tetanus toxin fragment C (CEA-DOM) or the Fc portion of IgG1 (CEA-FcIgG) were identified as highly immunogenic and elicited significant CEA-specific antibody and CD8+ T cell responses. CEA.tg mice were protected from tumor growth on challenge with MC38-CEA tumor cells only when immunized with repeated injections of plasmid pV1J/CEA-DOM followed by Ad/CEA-DOM. Depletion of T-regulatory cells resulted in an increased immune response and antitumor effect with DNA plus adenovirus immunization. In addition, this protective effect was abrogated if the NK, CD4+, or CD8+ cell population from immunized mice was depleted before tumor challenge. Passive transfer studies demonstrated that CD4+ and CD8+ T cells and antibodies contributed to the antitumor effect, thus suggesting that a genetic vaccine based on the use of plasmid DNA and adenoviral vectors encoding CEA fused to immunoenhancing sequences augments CEA-specific immune responses and effectively protects from tumor development.

Adenoviridae↗
PubMed2005

caBIONet--A .NET wrapper to access and process genomic data stored at the National Cancer Institute's Center for Bioinformatics databases.

MOTIVATION: The National Cancer Institute's Center for Bioinformatics (NCICB) has developed a Java based data management and information system called caCORE. One component of this software suite is the object oriented API (caBIO) used to access the rich biological datasets collected at the NCI. This API can access the data using native Java classes, SOAP requests or HTTP calls. Non-Java based clients wanting to use this API have to use the SOAP or HTTP interfaces with the data being returned from the NCI servers as an XML data stream. Although the XML can be read and manipulated using DOM or SAX parsers, one loses the convenience and usability of an object oriented programming paradigm. caBIONet is a set of .NET wrapper classes (managers, genes, chromosomes, sequences, etc.) capable of serializing the XML data stream into local .NET objects. The software is able to search NCICB databases and provide local objects representing the data that can be manipulated and used by other .NET programs. The software was written in C# and compiled as a .NET DLL.

Chromosome Mapping↗
PubMed2000

Interaction among SOX10, PAX3 and MITF, three genes altered in Waardenburg syndrome.

Waardenburg syndrome (WS) is an autosomal dominant disorder with an incidence of 1 in 40 000 that manifests with sensorineural deafness and pigmentation defects. It is classified into four types depending on the presence or absence of additional symptoms. WS1 and WS3 are due to mutations in the PAX3 gene whereas some WS2 cases are associated with mutations in the microphthalmia-associated transcription factor (MITF) gene. The WS4 phenotype can result from mutations in the endothelin-B receptor gene (EDNRB), in the gene for its ligand, endothelin-3 (EDN3), or in the SOX10 gene. PAX3 has been shown to regulate MITF gene expression. The recent implication of SOX10 in WS4 prompted us to test whether this transcription factor, known to cooperate in vitro with PAX3, is also able to regulate expression from the MITF promoter. Here we show that SOX10, in synergy with PAX3, strongly activates MITF expression in transfection assays. Analyses revealed that PAX3 and SOX10 interact directly by binding to a proximal region of the MITF promoter containing binding sites for both factors. Moreover, SOX10 or PAX3 mutant proteins fail to transactivate this promoter, providing further evidence that the two genes act in concert to directly regulate expression of MITF. In situ hybridization experiments carried out in the dominant megacolon (DOM:) mouse, confirmed that SOX10 dysfunction impairs MITF: expression as well as melanocytic development and survival. These experiments, which demonstrate an interaction between three of the genes that are altered in WS, could explain the auditory-pigmentary symptoms of this disease.

Animals↗
PubMedDate not supplied

Cross-reactivity of amphetamine analogues with Roche Abuscreen radioimmunoassay reagents.

Cross-reactivity of amphetamine analogues with the Abuscreen amphetamine radioimmunoassay reagents was determined for both the standard and high specificity antibody systems. Compounds tested included 2-methoxyamphetamine, 4-hydroxymethamphetamine, 2,5-dimethoxyamphetamine (DMA), 4-bromo-2,5-dimethoxyamphetamine (DOB), 4-bromo-2,5-dimethoxy-beta-phenethylamine (BDMPEA), 3,4,5-trimethoxyamphetamine (TMA), 3,4-methylenedioxyamphetamine (MDA), N,N-dimethyl-3,4-methylenedioxyamphetamine and N-hydroxy-3,4-methylenedioxyamphetamine (N-OH MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), 2,5-dimethoxy-4-ethylamphetamine, 2,5-dimethoxy-4-methylamphetamine (DOM), and 3,4,5-trimethoxyphenethylamine (mescaline). Blank negative reference material was spiked with 1,000 to 100,000 ng/mL of the amphetamine analogue and used as sample in the assays. MDA was the only analogue that showed cross reactivity equal to or greater than that of amphetamine. None of the other analogue compounds demonstrated a positive result at even the highest concentration; however several showed depressed counts at various concentration levels.

3,4-Methylenedioxyamphetamine↗
PubMed2004

Breast cancer and the brain: a neurodevelopmental hypothesis to explain the opposing effects of caloric deprivation during the Dutch famine of 1944-1945 on breast cancer and its risk factors.

Most studies on calorie deprivation and cancer risk in rodents show reductions in tumor occurrence. However, the few human studies on calorie restriction are conflicting. An overview is given of results in the DOM (diagnostic onderzoek mammacarcinoom) cohorts among women exposed to the Dutch Famine of 1944-1945. Opposing effects were found on risk factors (shortening of leg length, later menarche, and earlier menopause), whereas urinary estrogens and plasma insulin-like growth factor (IGF)-1 and IGF binding protein-3 were increased, as was breast cancer itself. Exposure between 2 and 10 y old was an unexpected window of susceptibility to the effects of calorie deprivation. The effects of famine exposure were most clearly seen in women who never gave birth. These opposing observations can be explained by a neurodevelopmental hypothesis on set-point shifts at the level of the diencephalons/hypothalamus, either directly or from rebound effects. Such a mechanism reflects old evolutionary adaptation systems in lower and higher organisms to cope with periods of stress and famine by adjusting, for example, reproductive functions. These effects in exposed women may later also affect their unexposed offspring. This hypothesis provides several testable, hormone-mediated corollaries on the relationships between the role of calories in a Westernized lifestyle and human cancer risk. The underlying developmental perspective, as opposed to a risk factor approach, can explain why certain ages, even before breast development, are especially sensitive to effects of large fluctuations in calories. The observations presented may have implications for preventive strategies such as promoting moderation of calorie intake to curb cancer risks.

Adult↗
PubMed2006

Structural basis of the Methanothermobacter thermautotrophicus MCM helicase activity.

The MCM complex from the archaeon Methanother-mobacter thermautotrophicus is a model for the eukaryotic MCM2-7 helicase. We present electron-microscopy single-particle reconstructions of a DNA treated M.thermautotrophicus MCM sample and a ADP.AlF(x) treated sample, respectively assembling as double hexamers and double heptamers. The electron-density maps display an unexpected asymmetry between the two rings, suggesting that large conformational changes can occur within the complex. The structure of the MCM N-terminal domain, as well as the AAA+ and the C-terminal HTH dom-ains of ZraR can be fitted into the reconstructions. Distinct configurations can be modelled for the AAA+ and the HTH domains, suggesting the nature of the conformational change within the complex. The pre-sensor 1 and the helix 2 insertions, important for the activity, can be located pointing towards the centre of the channel in the presence of DNA. We propose a mechanistic model for the helicase activity, based on a ligand-controlled rotation of the AAA+ subunits.

Adenosine Triphosphatases↗
PubMed1998

Hypertension, antihypertensive drugs, and mortality from cancer among women.

OBJECTIVES: To investigate whether hypertension and the use of antihypertensive drugs are associated with mortality from cancer among women. DESIGN: A prospective study of 11075 women living in Utrecht, the Netherlands, aged 50-65 years at enrolment in a breast cancer screening project (DOM cohort). Women attended screening rounds between 1974 and 1985, during which blood pressure measurements were taken and information on drug use and smoking was ascertained. Since 1974 (median follow-up time 19 years) information on cause of death has been obtained from the patient's general practitioner. Hypertension was defined as a systolic blood pressure > 160 mmHg, a diastolic blood pressure > 95 mmHg, or current use of antihypertensive drugs. Cox regression analysis was used to investigate the association between hypertension (treated and untreated) and total and site-specific mortalities from cancer. The influences of systolic and diastolic blood pressures per 10 mmHg increase for women not using antihypertensive drugs were evaluated. Analyses were adjusted for age, smoking, and body mass index. RESULTS: In total, 704 women died of cancer and 1633 women left the study area. Hypertensive women had a greater than normal (not statistically significant) total risk of mortality from cancer [hazard ratio (HR) 1.10, 95% confidence interval (CI) 0.93-1.31]. Risks for treated and untreated hypertensive women were similar. Cancer-site-specific analyses revealed a statistically significantly lower mortality from gastrointestinal cancers among untreated hypertensive women (HR 0.64, 95% CI 0.44-0.93), whereas mortality from lung cancer was more prevalent among these women (HR 2.50, 95% CI 1.37-4.59). Mortality from lymphatic and hematopoietic cancers for drug-treated hypertensive women was greater than normal (HR 2.11, 95% CI 1.04-4.28), as was mortality from cancers of the uterus, cervix, and ovary (HR 1.80, 95% CI 1.00-3.26). CONCLUSION: These results support the hypothesis that, if there is a link between blood pressure and cancer, it is likely to be positive and relatively small (+10%); and applies also to nondrug-treated women. The relation may apply for some types of cancer, but not for others.

Aged↗
PubMed2001

Human hamstring muscles adapt to eccentric exercise by changing optimum length.

PURPOSE: It is now established that unaccustomed eccentric exercise leads to muscle fiber damage and to delayed-onset muscle soreness (DOMS) in the days after exercise. However, a second bout of eccentric exercise, a week after the first, produces much less damage and soreness. The purpose of this study was to provide evidence from muscle mechanical properties of a proposed mechanism for this training effect in human hamstring muscles. METHODS: The eccentric exercise involved 12 sets of 6 repetition "hamstring lowers," performed on specially designed equipment. Hamstring angle-torque curves were constructed for each of 10 subjects (8 male and 2 female) while they performed maximum voluntary knee extension and flexion movements on an isokinetic dynamometer. Testing sessions were performed over the week before eccentric exercise, immediately post exercise, and daily, up to 8 d post exercise. Subject soreness ratings and leg girth measurements were also made post exercise. Six subjects performed a second bout of eccentric exercise, 8 d after the first, and measurements were continued up to 10 d beyond that. RESULTS: There was a significant shift in the optimum angle for torque generation (Lo), to longer muscle lengths immediately post exercise (7.7 degrees +/- 2.1 degrees, P < 0.01), indicating an increase in series compliance within some muscle fibers. Subsequent measurements showed increases in leg girth and some muscle soreness, suggesting muscle damage. The shift in Lo persisted, even after other injury parameters had returned to normal, consistent with a training effect. Subjects also showed fewer signs of muscle damage after the second exercise bout. CONCLUSION: This is the first study to show a sustained shift in optimum angle of human muscle as a protective strategy against injury from eccentric exercise. Implications of this work for athletes, particularly those prone to hamstring strains are discussed.

Adaptation, Physiological↗
PubMed2002

Electro-membrane microcurrent therapy reduces signs and symptoms of muscle damage.

PURPOSE: Delayed onset muscle soreness (DOMS) occurs after unaccustomed physical activity or competitive sport, resulting in stiff, painful muscles with impaired function. Acustat electro-membrane microcurrent therapy has been used to treat postoperative pain and soft tissue injury; however, its efficacy in reducing symptoms of muscle damage is not known. METHODS: Thirty healthy men were recruited for a double-blind, placebo-controlled trial. The muscles of their nondominant arms were damaged using an eccentric-exercise protocol. Subjects were then randomly assigned to treatment with either Acustat or a matching placebo membrane for 96 h and monitored for a total of 168 h. RESULTS: Subjects in both groups experienced severe pain and swelling of the elbow flexors after the eccentric exercise. After 24 h, the elbow joint angle of the placebo group had increased significantly more than those in the Acustat group (13.7 +/- 8.9 degrees vs 7.5 +/- 5.5 degrees; placebo vs Acustat, P < 0.05), possibly as a consequence of the elbow flexor muscles shortening. For the first 48 h after exercise, maximum voluntary contraction of the elbow flexor muscles was significantly impaired in the placebo group by up to 25% (P < 0.05), whereas muscle function was unchanged in the Acustat group. Peak plasma creatine kinase activity was also lower in the Acustat group (peak = 777 +/- 1438 U.L-1) versus the placebo group (peak = 1918 +/- 2067 U.L-1; (P < 0.05). The membranes were well tolerated by the subjects in both groups without any adverse effects. CONCLUSION: These data show that treatment of muscle damage with Acustat electro-membrane microcurrent therapy reduces the severity of the symptoms. The mechanisms of action are unknown but are likely related to maintenance of intracellular Ca2+ homeostasis after muscle damaging exercise.

Adult↗
PubMed2004

Optimal positioning for a dental operating microscope during nonsurgical endodontics.

The most comfortable positioning for a dental operating microscope (DOM) during nonsurgical endodontics for operators was investigated. Operators were categorized into 3 groups according to height. We recorded the time taken to obtain magnified images, and the angles of binoculars, microscope body, and the mirror to floor surfaces. For the group of shorter operators, observations were also made with an angled optics or a short objective lens (200 mm). It took longer to observe the mandibular molars than maxillary in every group. Although the differences in angles among each group were not remarkable for maxillary observation, we confirmed significant differences for the mandibular. Shorter operators had to adopt a strained position for mandibular observation in a standard setup but were more comfortable using angled optics or a short objective lens. By understanding the proper position, operators could learn to perform microendodontics more efficiently.

Body Height↗
PubMed2001

Detection of palm fruit lipids in archaeological pottery from Qasr Ibrim, Egyptian Nubia.

In modern times, the trees of the palm family have been of great economic and social importance to the people in Egypt, as in other parts of the world. There are various species of palm and although different parts of the tree can be used, the fruit are of great value. In antiquity, it is expected that the palm fruit would also have been of great importance to people in the region. The chemical analysis of absorbed residues in archaeological pottery is well established, and through the investigation of ceramic vessels (via gas chromatography, gas chromatography-mass spectrometry and gas chromatography-combustion-isotope ratio mass spectrometry) saturated carboxylic acids in the range C12 to C18 have been detected (with an unusually high abundance of C12) from vessels from the Nubian site of Qasr Ibrim. This is mirrored in the saturated fatty acid distributions detected from the kernels of modern and ancient date palm (Phoenix dactylifera L.) and dom palm (Hyphaena thebaica (L.) Mart.). Mixing in some of the vessels of the palm fruit with another lipid source is indicated through the delta13C values. These results provide the first direct evidence for the exploitation of palm fruit in antiquity and the use of pottery vessels in its processing.

Archaeology↗
PubMed1987

An evaluation of the interactions between freshwater pulmonate snail hosts of human schistosomes and macrophytes.

An account is given of a laboratory investigation designed to evaluate the extent to which the freshwater pulmonate snail Biomphalaria glabrata (Say) can utilize various species of aquatic plants, mainly macrophytes, when presented in the following forms over different time scales: normal plants; dried plant material; homogenized plant material in calcium alginate matrices; water-soluble filtrates of plant homogenates in the medium. The following propositions, derived from the theory of phased coevolution of components of the module consisting of the epiphytic bacteria, algae, snails and macrophytes, are evaluated on the basis of the present results and others including those obtained in this laboratory. That as the snails had become specialized to exploit surface communities of epiphytic algae, decaying plant material and dissolved organic matter (DOM) early in their evolutionary history they would continue to exploit these resources when they later become associated with aquatic macrophytes. That pulmonate snails would tend to be feeding generalists capable of adapting to food of varying chemical composition, given sufficient time, provided it was sufficiently small or flaccid. That although macrophytes and snails show a strong positive relationship, the living macrophyte tissue would be little used by the snails. That the hard outer envelope, inherited from their terrestrial ancestors, would remain as the major defence mechanism of aquatic macrophytes against attack by snails and other aquatic invertebrates. That aquatic macrophytes would invest little in the nutrient deficiency strategy to reduce attack by invertebrates such as snails. That truly aquatic submerged macrophytes would not possess secondary plant compounds (SPC) that would be molluscicidal. Emergent parts of subaquatic or aquatic plants might be expected to be better sources of SPC with molluscicidal factors than submerged aquatic plants. Species of epiphytic or planktonic algae might be better sources of SPC with molluscicidal effects than aquatic macrophytes. That the strategies developed by pulmonate snails for obtaining their energy supplies would not be conducive to rapid speciation. The analysis of the present and other related results supports these propositions. Predictions based on the theory of mutualism involving the snails, macrophytes and other components of the module also receive some support from an analysis of the present results. The additional empirical work that could be undertaken to test this theory is briefly discussed.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed2000

Development of a chromosome-plasmid balanced lethal system for Lactobacillus acidophilus with thyA gene as selective marker.

A chromosome-plasmid balanced lethal gene delivery system for Lactobacillus acidophilus based on the thyA gene was developed. The selected L. acidophilus DOM La strain carries a mutated thyA gene and has an obligate requirement for thymidine. This strain can be used as a host for the constructed shuttle vector pFXL03, lacking antibiotic-resistant markers but having the wild-type thyA gene from L. casei which complements the thyA chromosomal mutation. The vector also contains the replicon region from plasmid pUC19 and that of the Lactococcus plasmid pWV01, which allows the transfer between Escherichia coli, L. casei and L. acidophilus. Eight unique restriction sites (i.e., PstI, HindIII, SphI, SalI, AccI, XbaI, KpnI and SacI) are available for cloning. After 40-time transfers in modified MRS medium, no plasmid loss was observed. The vector pFXL03 is potentially useful as a food-grade vaccine delivery system for L. acidophilus.

Drug Delivery Systems↗
PubMed2005

Professor Peter Choyce: an early pioneer of intraocular lenses and corneal/refractive surgery.

Professor Peter Choyce, FRCS, DOMS, MS, was one of the pioneers of intraocular lens implant surgery. He developed an interest in artificial lens implantation following cataract surgery, a procedure that was widely criticized by the ophthalmic establishment in the UK, Europe, North America and other countries. Owing to the opposition to the intraocular lenses, Peter Choyce together with Sir Harold Ridley co-founded the International Intraocular Implant Club in 1966, which was responsible for the gradual acceptance of artificial lens implantation. Peter Choyce developed several models of intraocular lens, but did not patent the majority of them. The Choyce Mark IX, manufactured by Rayner Intraocular Lenses, became the first US Food and Drug Administration-approved intraocular lens in 1981. A review of Peter Choyce's record confirms a significant number of original innovations in the field of anterior segment surgery, including many procedures taken for granted today, but not associated with his name. These include early work on both kerato- and intraocular lens-refractive procedures, keratoprosthesis, pioneering paediatric implant procedures and others. Unfortunately his tenacious adherence to anterior chamber lens technology, while in general clinically sound, caused many to question his influence and hence he remained poorly understood even until after his death. He passed away on 8 August 2001 after a long fight with colon cancer. In this article, we provide evidence and elaborate Peter Choyce's accomplishments, which places him as one of the most innovative ophthalmologist in his surgical field in the twentieth century.

Cornea↗
PubMed2005

Methane fluxes in permafrost habitats of the Lena Delta: effects of microbial community structure and organic matter quality.

For the understanding and assessment of recent and future carbon dynamics of arctic permafrost soils the processes of CH(4) production and oxidation, the community structure and the quality of dissolved organic matter (DOM) were studied in two soils of a polygonal tundra. Activities of methanogens and methanotrophs differed significantly in their rates and distribution patterns among the two investigated profiles. Community structure analysis showed similarities between both soils for ester-linked phospholipid fatty acids (PLFAs) and differences in the fraction of unsaponifiable PLFAs and phospholipid ether lipids. Furthermore, a shift of the overall composition of the microbiota with depth at both sites was indicated by an increasing portion of iso- and anteiso-branched fatty acids related to the amount of straight-chain fatty acids. Although permafrost soils represent a large carbon pool, it was shown that the reduced quality of organic matter leads to a substrate limitation of the microbial metabolism. It can be concluded from our and previous findings first that microbial communities in the active layer of an Arctic polygon tundra are composed by members of all three domains of life, with a total biomass comparable to temperate soil ecosystems, and second that these microorganisms are well adapted to the extreme temperature gradient of their environment.

Archaea↗
PubMed1994

Species-dependent functional properties of non-NMDA receptors expressed in Xenopus laevis oocytes injected with mammalian and avian brain mRNA.

1. Species-dependent variation in the functional properties of non-NMDA receptors was investigated by intracellular recording in Xenopus laevis oocytes injected with rat, chick and calf brain mRNA. 2. In all mRNA-injected oocytes, kainic acid (KA), domoic acid (Dom) and 5-bromowillardiine (BrW) evoked large, maintained membrane currents, in contrast to the smaller, desensitizing responses elicited by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), quisqualic acid (QA) and L-glutamic acid (L-Glu). Dose-response curves for KA in oocytes injected with calf (EC50 = 96.4 +/- 12.3 microM; mean +/- s.e. mean), chick (87.0 +/- 8.9 microM) or rat (88.7 +/- 4.3 microM) brain mRNA were similar. 3. Current-voltage (I-V) relationships determined with KA inwardly rectified in oocytes injected with calf or chick mRNA; whereas, outward rectification was observed in oocytes injected with rat brain mRNA. 4. In oocytes injected with rat brain mRNA, AMPA antagonized responses evoked by KA in a competitive manner. The absolute amplitudes of KA and AMPA responses in the same oocytes were significantly correlated, which is consistent with both agonists acting on the same receptor-ionophore complex. 5. In contrast, in oocytes injected with calf or chick brain mRNA, AMPA (QA and L-Glu) antagonized the response evoked by KA in a non-competitive manner. The response amplitudes of KA compared to AMPA, QA or L-Glu in the same oocytes were not correlated suggesting discrete receptor-ionophores. 6. This study favours the existence of distinct non-NMDA receptor subtypes that are equi-sensitive to KA. The expressed receptors from different species of mRNA may be distinguished by their voltage sensitivities and the type of antagonism exerted by AMPA on KA-activated responses. Our observations may reflect further heterogeneity of non-NMDA receptors in the central nervous system of different vertebrate species.

Animals↗
PubMed1998

The effect of passive stretching on delayed onset muscle soreness, and other detrimental effects following eccentric exercise.

The aim of this study was to measure if passive stretching would influence delayed onset muscle soreness (DOMS), dynamic muscle strength, plasma creatine kinase concentration (CK) and the ratio of phosphocreatine to inorganic phosphate (PCr/P(i)) following eccentric exercise. Seven healthy untrained women, 28-46 years old, performed eccentric exercise with the right m. quadriceps in an isokinetic dynamometer (Biodex, angle velocity: 60 degrees.s-1) until exhaustion, in two different experiments, with an interval of 13-23 months. In both experiments the PCr/P(i) ratio, dynamic muscle strength, CK and muscle pain were measured before the eccentric exercise (day 0) and the following 7 d. In the second experiment daily passive stretching (3 times of 30 s duration, with a pause of 30 s in between) of m. quadriceps was included in the protocol. The stretching was performed before and immediately after the eccentric exercise at day 0, and before measurements of the dependent variables daily for the following 7 d. The eccentric exercise alone led to significant decreases in PCr/P(i) ratio (P < 0.001) and muscle strength (P < 0.001), and an increase in CK concentration (P < 0.01). All subjects reported pain in the right m. quadriceps with a peak 48 h after exercise. There was no difference in the reported variables between experiments one and two. It is concluded that passive stretching did not have any significant influence on increased plasma-CK, muscle pain, muscle strength and the PCr/P(i) ratio, indicating that passive stretching after eccentric exercise cannot prevent secondary pathological alterations.

Adult↗