Prospects for use of iodine-free radiopaques for cholecystocholangiography

By using the compounds tantalum and lanthanum, a new radiopaque substance (ROS) was synthesized at the Institute for Chemistry of Solids, Urals Branch of the Russian Academy of Sciences. Experiments on laboratory animals provided evidence that there was no acute or chronic toxicity of helium-based lanthanum orthotantalate. The specific radiopaque properties of the agent versus iodine- and barium-containing ROSs were studied in in vitro and in vivo experiments. The radiodiagnostic properties and locally irritant effect of the test ROS versus iodine-containing agents were evaluated in dogs at cholecystocholangiography.     

Risk factors for rejection for morphological reasons of heart valves for transplantation

The study aim was to identify risk factors for morphological rejection of aortic and pulmonary valves for transplantation that could be used to optimize donor selection. The files of all Dutch heart valve donors, donating in a 2.5 years period, whose hearts were processed at Heart Valve Bank Rotterdam, were reviewed for all factors that could be relevant for valve rejection and related to outcome of morphological assessment of the valves. Valves were retrieved from 813 deceased Dutch donors, 24.1% also donating organs. For 797 aortic and 767 pulmonary valves, who met retrieval criteria, morphological assessment was done. 69.5% of aortic and 37.5% of pulmonary valves were considered unsuitable for transplantation at morphological assessment. Backward stepwise multivariate logistic regression analysis, showed age, cardiac cause of death, cerebrovascular accident as cause of death or in medical history, and number of cardiovascular risk factors in a donor to be independent risk factors for morphological rejection of aortic valves. Age, sex, weight >100 kg and ruptured aortic aneurysm as cause of death were independent risk factors for morphological rejection of pulmonary valves. Being an organ donor was an independent predictor of morphological approval of aortic and pulmonary valves, while hypertension was an independent predictor for morphological approval of aortic valves. Thus, independent factors were identified that are associated with morphological rejection of aortic and pulmonary valves for transplantation, and that could be used to optimize donor selection by preventing unnecessary retrievals, limiting costs, while improving yield per donor with minimal compromise for availability.     

Standards for Approval of Hospitals for Graduate Training

The assessment program for approval of hospitals for training in specialties has been augmented by survey visits. Surveyors for the Canadian Council on Hospital Accreditation have acted on behalf of the Royal College of Physicians and Surgeons of Canada and have assisted in the formulation of standard criteria for evaluation of teaching programs. Areas where considerable variation in program exists and where further standardization might profitably be developed are suggested. It is recommended that periodic on-the-site visits be made to all specialty training hospitals. Standards should be further elucidated to guide teachers, students and Royal College surveyors alike. It is emphasized that standards cannot be used to assess the calibre of teaching and learning, but can be employed to assist in evaluating the teachings and learning media that are likely to produce good (or poor) results.     

Method for epitope selection of antisera for immunohistology

Polyclonal anti-laminin serum was affinity-purified on paraformaldehyde-fixed laminin on a nitrocellulose filter. The purified antibodies were tested for their specificity in immunohistological stainings on frozen sections of paraformaldehyde-fixed tissue. As compared to the initial polyclonal serum, the purified antibodies increased the specificity of antigen detection, since all background caused by nonspecific reactions was eliminated. This technique promises to be very useful for immunohistological analysis using light and electron microscopy.     

Cercariometry for detection of transmission sites for schistosomiasis

Cercariometry provided information on diurnal fluctuation, seasonal and spatial distribution of cercariae in the suitable natural water bodies. There was an apparent mismatch between the results of cercariometry and snail sampling. Water, which cercariometry showed to contain cercariae was potentially infective, although the resultant worm load of sentinel rodents may not bear a linear relationship with cercarial density. Cercariometry has some weakness in practices and analysis of data, however, it provides the valuable information on the active transmission sites of schistosomiasis.     

Algorithms for mapping of mRNA targets for microRNA

MicroRNAs (miRNAs) are involved in human health and disease as endogenous suppressors of the translation of coding genes. At this early point of time in miRNA biology, it is important to identify specific cognate mRNA targets for miRNA. Investigation of the significance of miRNAs in disease processes relies on algorithms that hypothetically link specific miRNAs to their putative target genes. The development of such algorithms represents a hot area of research in biomedical informatics. Lack of biological data linking specific miRNAs to their respective mRNA targets represents the most serious limitation at this time. This article presents a concise review addressing the most popular concepts underlying state-of-the-art algorithms and principles aimed at target mapping for specific miRNAs. Strategies for improvement of the current bioinformatics tools and effective approaches for biological validation are discussed.     

Equipment for Simultaneous Processing of Groups of Slides for Radioautography

The equipment consists of commercially available 5-slide plastic grips (obtained from Lipshaw Manufacturing Co., 7446 Central Ave., Detroit, Mich. 48210) and specially designed lightproof exposure boxes to hold the grips. After deparaffinization and hydration, slides are inserted into the grips, dipped in liquid emulsion, drained, and sealed in the exposure box. After exposure, the 5-slide units are removed from the box (in darkness) and inserted into specially made Plexiglas racks that hold four 5-slide units (20 slides). Photographic developing, fixing, and washing are performed by immersing the 20-slide group in fluids contained in 30-slide glass staining dishes (Lipshaw No. 122 dishes). The equipment simplifies darkroom operations and provides means for standardizing the preparation of radioautographs.     

Assessment of Needs for Plasma for Fractionation in Europe

In the early 1990s, a series of outbreaks of hepatitis C (HCV) infections clustering among recipients of certain lots of plasma-derived medicinal products (PDMP) alarmed regulatory authorities, manufacturers and the public alike. Also, a few episodes of Hepatitis A (HAV) infections occurred in haemophiliacs receiving solvent-detergent-treated factor VIII concentrates. Thus, several measures were brought into effect to reestablish the safety of the incriminated products and to further increase the margin of safety of PDMP in general. Therefore, intramuscular immunoglobulins had to be free of HCV RNA as shown by nucleic acid amplification technology (NAT) in the final products. Furthermore, the manufacturing process of PDMP had to be validated for both viral inactivation and elimination. Finally, HCV-NAT was to be standardised and implemented as a validated test of plasma pool samples.In 1994, a joint meeting of EPFA, EAPPI and Regulatory Authorities was held in Brussels to outline the state of the art and to delineate the actions to be taken. Five years later, in 1999, the incidence rates of HIV, HBV and HCV in unpaid blood donors have been minimized, especially in European countries. With probabilities for window period donations as low as 0.6 in 1 million for both HIV and HCV and 2.1 in 1 million for HBV in Switzerland, labile blood products have reached extreme, but not absolute safety. The introduction of HCV-NAT roughly doubles this safety resulting in a 1 in 3 million probability of a window donation.Concomittantly, extensive viral validation studies document effective inactivation and removal of viruses in PDMP. The demonstrated margins of safety, expressed as logarithmical reduction factors (LRF), range from 4 to over 20 log(10), depending on product, virus, and inactivation procedure used. Further progress to even safer PDMP shall be acomplished by consolidating the GMP processes, abandoning of obsolete requirements and harmonising national regulations within Europe. Before introducing new measures for additional agents such as HAV or Parvovirus B 19, gains and risks and even potential new threats have to be carefully assessed. Alternative efforts for the safeguard of patients, e.g. vaccination for HAV, need to be balanced against the risks of changing established and validated manufacturing procedures of PDMP with long-lasting safety records.     

Nonlinear electrophoresis for purification of soil DNA for metagenomics

Purification of microbial DNA from soil is challenging due to the co-extraction of humic acids and associated phenolic compounds that inhibit subsequent cloning, amplification or sequencing. Removal of these contaminants is critical for the success of metagenomic library construction and high-throughput sequencing of extracted DNA. Using three different composite soil samples, we compared a novel DNA purification technique using nonlinear electrophoresis on the synchronous coefficient of drag alteration (SCODA) instrument with alternate purification methods such as direct current (DC) agarose gel electrophoresis followed by gel filtration or anion exchange chromatography, Wizard DNA Clean-Up System, and the PowerSoil DNA Isolation kit. Both nonlinear and DC electrophoresis were effective at retrieving high-molecular weight DNA with high purity, suitable for construction of large-insert libraries. The PowerSoil DNA Isolation kit and the nonlinear electrophoresis had high recovery of high purity DNA suitable for sequencing purposes. All methods demonstrated high consistency in the bacterial community profiles generated from the DNA extracts. Nonlinear electrophoresis using the SCODA instrument was the ideal methodology for the preparation of soil DNA samples suitable for both high-throughput sequencing and large-insert cloning applications.     

Identification of QTL for agronomic traits of importance for olive breeding

Development of methodologies for early selection is one of the most important goals of olive breeding programs at present. In this context, the identification of molecular markers associated with beneficial alleles could allow the development of marker-assisted selection (MAS) strategies in olive breeding programs. Fruit-related and plant vigor traits, which are of key importance for olive selection and breeding, were analyzed during two seasons in a progeny derived from the cross ‘Picual’ × ‘Arbequina.’ Quantitative trait loci (QTL) analyses were performed using MapQTL 4.0. A total of 22 putative QTLs were identified in the map of ‘Arbequina.’ QTLs clustered in linkage groups (LG) 1, 10 and 17. QTLs for oil-related traits located in LG 1 and 10 explained around 20–30 % of the phenotypic variability depending on the season and the trait. QTL for moisture-related traits were detected in LG 1, 10 and 17, and QTLs for the ratio pulp to stone were identified in LG 10 and 17 explaining around 15–20 %. Interaction between QTLs for the same trait was investigated. The significance of these results was discussed considering the co-localization of QTLs and Pearson correlations among traits. Five additional QTLs were detected in the map of ‘Picual.’ Four of them clustered in LG 17 indicating the presence of a QTL for fruit weight explaining around 12.7–15.2 % of the variability. Additionally, a QTL for trunk diameter was detected in LG 14 explaining 16 % of the variation. These results represent an important step toward the application of MAS in olive breeding programs.     

Procedure for Magnification of Lethal Deletions of Genes for Ribosomal RNA

GENE duplications are recognized as a powerful background for evolution^1–3: thus, one of the genes maintains the function, the other accumulates mutations. Several cell functions, however, require multiple copies of the same gene^4–7; and these are, generally, tightly clustered^8–13. One might hence expect to find many mutations within blocks of duplicates; but in the cases tested, none were found^9,12,14. Nevertheless, the genes of the cluster evolve^15,16. I think that mechanisms like magnification are the basis for the “horizontal” evolution of tandemly duplicated genes¹⁶; a specific model has recently been proposed¹⁷. The mechanism I propose is as follows. Unequal or sister strand crossing over leads to the formation of partial deletions of the block or to blocks of increased redundancy^18,19. Eventually there is selection against the latter²⁰, while partially deleted blocks in the appropriate conditions can magnify: that is, can form a large number of new copies on those left in the block. This will lead to an increase of homogeneity which will be greater the lower the number of genes used as copies to rebuild the block. It seemed desirable to test whether deletions of genes for rRNA strong enough to be lethal were susceptible to magnification. This paper shows that this is feasible and strengthens a previous result by Atwood²¹.     

A checklist for ecological management of landscapes for conservation

The management of landscapes for biological conservation and ecologically sustainable natural resource use are crucial global issues. Research for over two decades has resulted in a large literature, yet there is little consensus on the applicability or even the existence of general principles or broad considerations that could guide landscape conservation. We assess six major themes in the ecology and conservation of landscapes. We identify 13 important issues that need to be considered in developing approaches to landscape conservation. They include recognizing the importance of landscape mosaics (including the integration of terrestrial and aquatic areas), recognizing interactions between vegetation cover and vegetation configuration, using an appropriate landscape conceptual model, maintaining the capacity to recover from disturbance and managing landscapes in an adaptive framework. These considerations are influenced by landscape context, species assemblages and management goals and do not translate directly into on-the-ground management guidelines but they should be recognized by researchers and resource managers when developing guidelines for specific cases. Two crucial overarching issues are: (i) a clearly articulated vision for landscape conservation and (ii) quantifiable objectives that offer unambiguous signposts for measuring progress.     

New technic for purification of lymphocytes for HLA typing

Many of the difficulties encountered in HLA typing derive from the separation of relatively pure cell suspensions for the lymphocytotoxicity test. We have used Immunomagnetic Beads (I.B.) coated with anti-CD8 or CD2 MAb for Class I positive cell selection and I.B. coated with anti-DR for Class II. With this method, compared to traditional techniques, we obtained high purity and viability of cell populations (about 90%), directly from whole blood in 25 minutes. Shorter incubation with antisera and complement was needed and the global time of tissue typing results decreased (150 min. versus 6 hours). There was no single discrepancy in the tissue typing results between I.B. and traditional methods.     

Semiautomatic regulator for the preparation of microelectrodes for electrophysiology

This paper describes an electronic device for making metallic microelectrodes to be used for electrophysiological experiments. These microelectrodes are endowed with a small tissue-electrode capacity and a high mechanical strength. The same microelectrodes are applicable for derivation of spikes as in the case of microstimulation of nervous structures during repeated penetrations.     

Simple Method for Repurification of Endotoxins for Biological Use

A method for obtaining highly purified endotoxin (lipopolysaccharide [LPS]) in a few hours by repurification of commercial or laboratory preparations was devised. It avoids the use of phenol, which is not suitable for phenol-soluble lipopolysaccharides nor for some industrial purposes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization mass spectrometry analysis confirmed the integrity of the purified LPSs. The purified products did not activate Toll-like receptor 2 (TLR2), nuclear oligomerization domain 1 (NOD1), or NOD2 but did activate TLR4. Applied to different lipopolysaccharides, the method also improved their mass spectra, thus facilitating their structural analysis.     

Novel procedures for derivatization of ribosomes for crystallographic studies

Undecagold and tetrairidium clusters have been used for the preparation of heavy-metal derivatives of ribosomal particles, necessary for the evaluation of phases in the x-ray structure determination of these large particles. To obtain specific binding, monofunctional reagents of the clusters were prepared and were covalently bound to free sulfhydryl groups on the surface of the ribosome. In addition, a mutant of Bacillus stearothermophilus which lacks one ribosomal protein (BL11) was grown. The heavy-atom clusters were covalently bound to isolated protein BL11, and the modified protein was consequently reconstituted into the mutated ribosomal subunits. Crystallographic data have been collected from crystals of native particles, from the mutated ones, and from the iridium- and gold-derivatized subunits. All these crystal forms are isomorphous within the experimental error.     

Methods of searching for markers for serological serum diagnosis of tumors

This review describes the most popular methods of searching for serological markers of tumors that are used in a clinical setting, as well as a comparison of their efficiency.     

Diagnostic Criteria for Differentiation of Pseudomonads Pathogenic for Man

To determine the most useful diagnostic characters for the differentiation of pseudomonads pathogenic for man, including Pseudomonas maltophilia (Alcaligenes bookeri), P. stutzeri (Bacillus denitrificans), P. pseudomallei (Malleomyces pseudomallei), and apyocyanogenic strains of P. aeruginosa, a comparative examination was made of 58 strains of these bacilli for their morphological and biochemical characteristics. It was concluded that the criteria for differentiating these bacteria are type of flagellation, growth on S S Agar, fluorescein production, oxidation of carbohydrates in the medium of Hugh and Leifson, nitrogen gas production, gluconate oxidation, gelatinase activity, urease activity, lysine decarboxylase activity, arginine dihydrolase activity, oxidase reaction, sensitivity to polymyxin, requirement for methionine as a growth factor, and assimilation of organic compounds as the sole source of carbon and energy.     

Utility of Polycation-Treated Filters for the Assay of Receptors for VIP

Abstract

This study examined the utility of four polycationic agents for treating glass fibre filters used in the receptor binding assay for vasoactive intestinal peptide (VIP). Polyethylenimine (PEI), polybrene, protamine and methylated bovine serum albumin proved satisfactory in terms of low filter binding of free radioligand and retention of membrane-bound radioligand. Their performance was superior or comparable to untreated Millipore EGWP cellulose acetate filters which we had previously utilized but which are no longer manufactured. The results with polycations indicate the importance of ionic interactions between filter, biological membranes and radioligand in determining the performance of a filtration assay for radioligand-receptor binding. At a practical level, PEI has the disadvantage of potential toxicity. The satisfactory performance of the other polycations indicates that they provide safer alternatives to PEI for filtration assay of the VIP receptor and possibly receptors for other basic ligands.     

Method for assessment of functional affinity of antibodies for live bacteria

A rapid and convenient method for assessment of functional affinity of antibodies against live bacteria is described. When a combination of immunomagnetic separation (IMS) with bioluminescent or fluorescent genetic labelling of the cells was employed, the method showed good correlation with plate count. However, the use of reporter bacteria allowed results to be obtained within 1 h compared with days using conventional methods. Due to its lower detection limit, the bioluminescent assay performed better than the fluorescent assay. Antibody affinities for Escherichia coli O157:H7 and Salmonella enteritidis were examined at different environmental conditions such as pH 3-7, temperature 4-25 degrees C, and sodium chloride concentrations 0-5% and compared with sensitivities of ELISA.