Preparation of compound-specific and group-specific antibodies to 7-methylguanine and related 7-alkylguanines and their use in immunoaffinity purification

The preparation and characteristics of compound-specific and group-specific antibodies against 7-alkylguanines (7-alkGua) are described. A compound-specific antibody against 7-methylguanine was prepared using a hapten bound to carrier protein through the N2 position. In a competitive enzyme-linked immunosorbent assay (ELISA) 7-methylguanine (7-MeGua) showed 50% inhibition (I50%) at 10 pmol/well at room temperature, but the inhibition was found to be 40 times better at 4 degrees C (I50% at 250 fmol/well). When the antibody was bound to protein A-Sepharose CL4B 7-MeGua was retained in immunoaffinity columns. A group-specific antibody to 7-alkGua was prepared using 7-(2-carboxyethyl)guanine (7-CEGua) bound to carrier protein via the carboxyl group. In a competitive ELISA, this antibody cross-reacted well with 7-CEGua, 7-ethylguanine (7-EtGua), 7-(2-hydroxyethyl)guanine (7-HOEtGua) and 7-(2',3'-dihydroxy)-propylguanine (7-DHPGua) and some inhibition was seen with 7-MeGua. Immunoaffinity columns prepared from this antibody retained a number of 7-alkGua of diverse structure. 7-EtGua in calf thymus DNA treated with diethylsulphate and ethylnitrosourea was isolated by immunoaffinity purification and quantified by HPLC-fluorescence. These results illustrate the potential of immunoaffinity purification for both individual DNA adducts and groups of adducts.     

Routine prenatal screening for congenital heart disease: what can be expected? A decision-analytic approach

OBJECTIVES: This study assessed the potential impact of fetal ultrasound screening on the number of newborns affected by cardiac anomalies. METHODS: A decision model was developed that included the prevalence and history of congenital heart disease, characteristics of ultrasound, risk of abortion, and attitude toward pregnancy termination. Probabilities were obtained with a literature survey; sensitivity analysis showed their influence on expected outcomes. RESULTS: Presently, screening programs may prevent the birth of approximately 1300 severely affected newborns per million second-trimester pregnancies. However, over 2000 terminations of pregnancy would be required, 750 of which would have ended in intrauterine death or spontaneous abortion. Further, 9900 false-positive screening results would occur, requiring referral. Only the sensitivity of routine screening and attitude toward termination of pregnancy appeared to influence the yield substantially. CONCLUSIONS: The impact of routine screening for congenital heart disease appeared relatively small. Further data may be required to fully assess the utility of prenatal screening.     

Current and emerging molecular diagnostic technologies applicable to bacterial food safety

There is an increasing need for rapid test methods to certify the quality and safety of food products. Current tests applied for the microbiological assessment of food products are based on standard approved culture-based isolation methods and can take several days to yield results. Nucleic acid diagnostic (NAD) tests for the identification of bacterial foodborne pathogens employing in vitro amplification technologies are capable of sensitive and specific detection of single or multiple pathogens in foods in a shorter timeframe than traditional methods. New developments in molecular biosensors have the potential to provide at-line bioanalysis, whereas microarray-based technologies may in the future be the NAD platforms of choice for multiple pathogen detection and identification. This article reviews current and emerging NAD platforms for foodborne bacterial pathogens that have the potential to impact food safety.     

Immobilized α-amylase from Bacillus licheniformis: a potential enzymic time—temperature integrator for thermal processing

Biphasic and nth-order models were tested as to their usefulness to fit experimental inactivation data of Bacillus licheniformis α-amylase, immobilized on glass beads, and were discussed with respect to their suitability to characterize the considered enzymic system as a time—temperature integrator (TTI) to evaluate heat processes. Both isothermal and non-isothermal inactivation experiments were carried out. Model (kinetic) parameters (rate constant k, activation energy E_A and reaction order n) were estimated using a non-linear regression procedure. The results obtained, especially the activation energy of about 293 kJ mole^–1, indicated a potential use of this system as a TTI for heating processes in the temperature range of 96–108°C.     

Characterization of the intracellular GLUT-4 compartment

Learned behaviors and tolerance to ethanol can be maintained by peripheral injection of arginine8-vasopressin (vasopressin) under conditions in which they would otherwise be lost. However, the sites of this action in the brain have not been clearly identified. Using a polyclonal antibody raised against Fos and Fos-like proteins, we have demonstrated increases in immunoreactive Fos and Fos-like proteins in the suprachiasmatic, supraoptic and paraventricular nuclei of the hypothalamus, and lesser increases in piriform cortex and amygdala, of the rat 2 h after a s.c. injection of vasopressin. Our results suggest that the exogenous vasopressin may exert its central action by activating a cellular immediate early gene in specific brain regions.     

Physical, Chemical and Sensory Changes in Irradiated Fresh Pork Packaged in Modified Atmosphere

The effects of irradiation dose (0, 0.5 and 1.0 kGy), headspace oxygen (0, 10 and 20% O₂, balance nitrogen) and storage temperature (5, 15 and 25°C) on the physical, chemical and sensory changes in fresh pork were studied using factorial design experiments. Irradiation in the absence of oxygen extended the sensory shelf life of pork from 9 to 26 days at 5°C and from & lt; 2 to 2 days at 25°C. Oxygen in the package headspace combined with irradiation adversely affected physical, chemical and sensory characteristics of the end product.     

Palatability of prerigor cooked boar meat

Cooking reduces odor intensity in boar meat but also may induce lipid oxidation unless the meat pH is above approximately 6.0. This research was designed to determine the feasibility of cooking boar meat in the prerigor state to overcome boar odor and lipid oxidation problems. Prerigor and postrigor triceps brachii muscle samples from 40 boars (20 Duroc and 20 Yorkshire) were cooked to 60 degrees C, frozen and stored at -20 degrees C, reheated in a 60 degrees C water bath for 1 h, and then subjected to pH, thiobarbituric acid (TBA), and sensory analyses. Boar odor intensity and skatole concentration in backfat samples were determined by olfactory test and HPLC, respectively. Cooked (initial cooking) prerigor meat was found to have higher (P < .05) pH and lower (P < .05) TBA values than comparable postrigor meat (6.44 vs 6.09 and 2.15 vs 3.23, respectively). Regression analysis indicated an inverse relationship between pH and TBA values (r = -.52; P < .01). No appreciable changes in TBA values were noted after frozen storage for 14 to 98 d, but reheating increased TBA values (P < .05) in both prerigor and postrigor samples (3.45 vs 4.32, respectively). Sensory evaluation scores indicated that prerigor cooked meat was less tender with more pronounced rancid flavor than postrigor cooked meat (P < .05), but panelists may have allowed the toughness of the prerigor samples to adversely affect their flavor scores. No difference in boar odor was detected between rigor states or breeds. Mean skatole concentration in backfat was .12 micrograms/g and no difference was detected between breeds.(ABSTRACT TRUNCATED AT 250 WORDS)     

The complex pathology of trinucleotide repeats

The expansion of trinucleotide repeat sequences has now been shown to be the underlying cause of at least ten human disorders. Unifying features among these diseases include the unstable behavior of the triplet repeat during germline transmission when the length of the repeat exceeds a critical value. However, the trinucleotide repeat disorders can be divided into two distinct groups. Type I disorders involve the expansion of CAG repeats, which encode an expanded polyglutamine, inserted into the open-reading frame of a gene that is usually quite broadly expressed. Recently, mouse models for type I disorders have been developed and the basis of pathology is under study, both in these models and through biochemical and cell biological approaches. The type II disorders involve repeat expansions in noncoding regions of genes. The mechanisms by which these repeat expansions lead to pathology may be quite diverse.