Extent of radial sarcomere coupling revealed in passively stretched cardiac myocytes

We report a simple method, the PinPoint assay, for detecting and identifying single-base variations (polymorphisms) at specific locations within DNA sequences. An oligonucleotide primer is annealed to the target DNA immediately upstream of the polymorphic site and is extended by a single base in the presence of all four dideoxynucleotide triphosphates and a thermostable DNA polymerase. The extension products are desalted, concentrated, and subjected to delayed-extraction MALDI-TOF mass spectrometry. The base at the polymorphic site is identified by the mass added onto the primer. Heterozygous targets produce two mass-resolved species that represent the addition of both bases complementary to those at the polymorphic site. The assay is suitable for double-stranded PCR products without purification or strand separation. More than one primer can be simultaneously extended and then mass-analyzed. The mass spectrometric method thus shows promise for high-volume diagnostic or genotyping applications.     

Evaluation of different fracture-mechanical J-integral initiation values with regard to their usability in the safety assessment of components

Determining fracture-mechanical material characteristic values on the basis of the J-integral is described and stipulated in a variety of standards and guidelines. The individual specifications differ in terms of procedure when determining the characteristic values and, therefore, also in terms of the meaningfulness of the results. This paper presents the different procedures, suggested in the course of the development of test methods in the field of elastic—plastic fracture mechanics, used to characterize crack initiation behaviour with regard to their features as material characteristic values and their usability in the safety assessment of components.     

Biodistribution of radiolabeled monoclonal antibody E48 IgG and F(ab'')2 in patients with head and neck cancer

The amphibian pronephros is fated to die during early development. Pronephric cells undergo apoptosis and their function is replaced by the mesonephros, which becomes the functional kidney of the adult frog. Tadpoles of the northern leopard frog, Rana pipiens, were inoculated with a Lucké tumour herpesvirus (LTV) preparation. Most of the animals developed typical Lucké renal carcinomas at metamorphosis. Fewer developed carcinomas of the pronephric cell type. A pronephric carcinoma, rescued from apoptosis by the herpesvirus, was harvested from a post-metamorphic frog. The tumour was judged to be pronephric by its anatomical location (in the anterior part of the body) and because both mesonephric kidneys were intact and tumour-free upon removal of the tumour mass. A tumour fragment was fixed for histological examination, which confirmed that the tissue was a renal carcinoma. A further fragment was subjected to short-term culture in order to produce metaphase cells for cytogenetical analysis. Based upon silverstained nucleolar organizing region numbers, 14 of 15 metaphase cells were estimated to have the diploid number (2N = 26) of chromosomes and a karyotype was constructed which did not appear to differ from that of normal cells. A single cell was estimated to be tetraploid (4N = 52). This is the first report of chromosomes of a pronephric Lucké carcinoma. LTV replicates only in tumour tissue maintained in the cold. Because the frog in this study had been maintained in the laboratory at 22 degrees C for about 10 months, no viruses would have been detectable with electron microscopy. However, the presence of Lucké herpesvirus DNA was detected in tumour homogenates by polymerase chain reaction amplification of a 1.2 kbp Hind III restriction fragment of the LTV DNA. The presence of LTV DNA provided assurance that the rescued pronephric tumour was indeed a Lucké carcinoma.     

The economic cost of a streptococcal tonsillitis episode

OBJECTIVE: To estimate the cost of a streptococcal tonsillitis episode from the data of a questionnaire. SETTING: Five primary health centres in the west of Sweden. PARTICIPANTS: 101 consecutive patients treated for streptococcal tonsillitis. MAIN OUTCOME MEASURE: The cost estimation included costs for physician visit and drug, travel costs to and from the primary health centre, cost of lost production resulting from the patient's or the guardian's absence from work for physician visit or sick-leave, and cost of telephone consultation with a physician or nurse. RESULTS: The period of illness was on average seven days, time to recovery after treatment five days, and the mean period of sick-leave 2.5 days. The total cost of a tonsillitis episode was about SEK 3,300 (385 USD). Of this sum, the cost for the antibiotic accounted for only 3% and loss of production for 75%. CONCLUSION: Differences in the cost of drugs only have a minor influence on the total cost, while factors causing loss of production, such as efficacy and side effects of the drug, have a greater influence. Economic evaluation of pharmaceuticals will be more relevant in the future, and in the search for the most effective treatment, cost effective studies will be integrated with clinical trials.     

Oxysterol incorporation into rat aorta resulting in elastin compositional changes

The incorporation of dietary cholestan-3β, 5α, 6β-triol (triol) into rat thoracic aortic tissue and changes in amino acid composition of the elastin were investigated to identify the cytotoxic properties of the triol. Weanling male Sprague-Dawley rats were fed the following diets for three months: (i) normal chow, (ii) normal chow with 1% (w/w) cholesterol added, or (iii) normal chow with 0.9% (w/w) cholesterol and 0.1% (w/w) triol added. Triol levels in the blood and in the thoracic aortic tissue were measured. Compositional changes of elastin were also determined. After three months on the triol-containing diet, triol was found in the thoracic aorta but was not detected in the blood. Amino acid analyses of the aortic tissue elastin revealed that the proline levels in the triol-fed animals were significantly greater than in the other two diet groups, while the elastin levels of leucine, aspartate, arginine, and phenylalanine decreased significantly. The mechanism for these observed changes induced by triol may reflect alternate splicing of elastin messenger ribonucleic acid (mRNA) resulting in structural changes in the elastin molecule. Dietary triol does contribute to tissue triol content and is associated with aortic elastin compositional changes. How these changes may contribute to the development of cardiovascular disease is not known.     

Comparison of Nonenzymatic Browning Kinetics in Spray-dried and Freeze-dried Carbohydrate-based Food Model Systems

ABSTRACT: Nonenzymatic browning (NEB) rates of amorphous, carbohydrate-based, freeze-dried and spray-dried food model systems containing L-lysine and D-xylose as reactants (5% w/w) were investigated at different temperatures (40 °C, 50 °C, 60 °C, 70 °C, 80 °C, 90 °C). Samples were exposed to various relative vapor pressure levels (11%, 23%, and 33%) to adjust water contents. Water sorption was determined gravimetrically, and data were modeled using the Brunauer-Emmett-Teller and Guggenheim-Anderson-deBoer equations. Glass transition, T_g, was observed by differential scanning calorimetry. NEB was followed spectrophotometrically. The surface structures of freeze-dried and spray-dried models were observed by scanning electron microscopy. The freeze-dried samples and the spray-dried samples showed different surface structures and slight differences in thermal behavior. Crystallization of component sugars in the freeze-dried samples was slightly more delayed than in spray-dried samples. The glass transition temperatures in spray-dried samples were higher than those of the freeze-dried samples at the same water activity. The temperature dependence of NEB rate in both systems followed the Arrhenius kinetics, but the activation energies were different. Williams-Landel-Ferry equation could be used to model the NEB kinetics in the freeze-dried system, but for the spray-dried system, negative constant values were not within the allowable range.     

Water sorption isotherms of freeze-dried milk products: applicability of linear and non-linear regression analysis in modelling

Summary  Water sorption of various freeze-dried milk products was modelled using several water sorption isotherm models, most of which proved to be applicable. Sorption models were fitted to experimental data using linear and non-linear regression analysis. Both methods gave almost the same prediction of water sorption when the model had a good fit. The GAB model was considered to be the most applicable in predicting water sorption in practical applications, as the use of one universal model is desirable. Time-dependent changes, e.g. lactose crystallization above glass transition, were taken into account in the water sorption modelling. Water contents and relative humidities which allowed changes in physico-chemical properties were not included in the modelling, because of unsteady amounts of sorbed water.