Dosimetry methods used in the studies of the effects of protons on primates: a review.

In a program to determine the acute and long-term effects of the space radiation environment, a series of primate irradiations was conducted from 1964 through 1969 by the USAF School of Aerospace Medicine. The animals were exposed total-body to protons, electrons, and X rays. The doses assigned were either body midline or body entrance geometrically averaged across the exposure field depending on the degree of penetration (total or partial body) of the radiation used. A 10-cm-diameter unit-density cylinder was used as an appropriate model for the animal in the determination of the body doses. The physical dosimetry methods used at the time of the irradiations are reviewed.     

Quality control parameters on a large dataset of regionally dissected human control brains for whole genome expression studies

We are building an open-access database of regional human brain expression designed to allow the genome-wide assessment of genetic variability on expression. Array and RNA sequencing technologies make assessment of genome-wide expression possible. Human brain tissue is a challenging source for this work because it can only be obtained several and variable hours post-mortem and after varying agonal states. These variables alter RNA integrity in a complex manner. In this report, we assess the effect of post-mortem delay, agonal state and age on gene expression, and the utility of pH and RNA integrity number as predictors of gene expression as measured on 1266 Affymetrix Exon Arrays. We assessed the accuracy of the array data using QuantiGene, as an independent non-PCR-based method. These quality control parameters will allow database users to assess data accuracy. We report that within the parameters of this study post-mortem delay, agonal state and age have little impact on array quality, array data are robust to variable RNA integrity, and brain pH has only a small effect on array performance. QuantiGene gave very similar expression profiles as array data. This study is the first step in our initiative to make human, regional brain expression freely available.     

Structural and performance characteristics of representative anion exchange resins used for weak partitioning chromatography

Weak partitioning chromatography (WPC) has been proposed for the purification of monoclonal antibodies using an anion exchange (AEX) resin to simultaneously remove both acidic and basic protein impurities. Despite potential advantages, the relationship between resin structure and WPC performance has not been evaluated systematically. In this work, we determine the structure of representative AEX resins (Fractogel® EMD TMAE HiCap, Q Sepharose FF, and POROS 50 HQ) using transmission electron microscopy and inverse size exclusion chromatography and characterize protein interactions while operating these resins under WPC conditions using two mAb monomers, a mAb dimer, mAb multimers, and BSA as model products and impurities. We determine the isocratic elution behavior of the weakly bound monomer and dimer species and the adsorptive and mass transfer properties of the strongly bound multimers and BSA by confocal laser scanning microscopy. The results show that for each resin, using the product K_p value as guidance, salt, and pH conditions can be found where mAb multimers and BSA are simultaneously removed. Isocratic elution and adsorption mechanisms are, however, different for each resin and for the different components. Under WPC conditions, the Fractogel resin exhibited very slow diffusion of both mAb monomer and dimer species but fast adsorption for both mAb multimers and BSA with high capacity for BSA, while the Sepharose resin, because of its small pore size, was unable to effectively remove mAb multimers. The POROS resin was instead able to bind both multimers and BSA effectively, while exhibiting a greater resolution of mAb monomer and dimer species. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:425–434, 2017     

Transformation ofBrassica oleracea L.: a critical review

Brassica oleracea is a highly polymorphic species encompassing a wide range of important vegetable and fodder crops. Gene transfer into cultivated forms of this species requires reproducible and efficient methods for genetic transformation and plant regeneration. In this review, we have collated the research experience on transformation ofB. oleracea to highlight the problems encountered. Most research effort has been directed at developingAgrobacterium-mediated transformation methods with relatively little emphasis to date on direct gene transfer techniques. Common procedures for the transformation ofB. oleracea have not emerged, due to the inherent variability between and amongst genotypes. Future progress would be facilitated by the use of genetically fixed material, such as double-haploid or inbred lines, to reduce variation of response within genotypes and would avoid the need for cultivar-specific transformation protocols if responsive lines amenable to crossing with cultivated forms could be identified. The principal difficulties relate to combining efficient plant regeneration with gene transfer. Methods that enhance bacterial virulence and increase the proportion of cells susceptible to transformation and competent for regeneration are discussed. Inefficient selection is a major cause of poor transformation frequencies inB. oleracea and has resulted in the regeneration of chimeric plants uponAgrobacterium tumefaciens-mediated transformation. Promising results have been obtained withAgrobacterium rhizogenes-mediated transformation but the impact of therol genes on flowering of primary transformants has not yet been fully assessed. Strategies to reduce the deleterious effects of therol genes on flowering are discussed. Few agronomically useful characters have been introduced, the majority of research having been confined to the introduction of marker and reporter genes; possible candidate genes are discussed.     

A critical review and a new proposal of karyotype asymmetry indices

In literature seven different methods of evaluating karyotype asymmetry – the TF%, the As K%, Stebbins' classification, the Rec and the Syi, the A₁ and the A₂, the DI, and the A – are used for the elucidation of phylogenetic relationships and taxonomic treatments within a particular group or taxon. The investigation of these seven methods reveals that the intervals used by Stebbins to separate the different types of karyotype asymmetry are very broad and only one quantitative parameter, the A₂ index, correctly describes the variation in chromosome length in a complement. A new asymmetry index (AI) is proposed to measure karyotype asymmetry and a new parameter, the CV_CI, is offered, that precisely assesses the relative variation in centromere position in a complement. The AI index, the CV_CI and the CV_CL (=A₂ × 100) have the potential to display even minor karyotypic variations. Thus, these three indices together increase the precision of results in comparison with other existing methods. All this has important consequences as regards the interpretation of the results of karyological studies.     

The demographic impact of breastfeeding: a critical review.

Demography has corrected the old wives tale that nursing mothers cannot become pregnant. Few realize the extent to which knowledge about the effects of breast feeding on birth intervals is based on tenous assumptions, biased observations, and widely divergent methodology. The purpose of this analysis is to provide a word of caution for those who are unaware that the major reasearch on lactation and postpartum ovulation is yet to be done. Data from which the demographic significance of lactation is estimated come primarily from 3 types of studies: 1) analysis of completed inter-birth intervals in populations assumed not to practice birth control; 2) surveys in which information on both breastfeeding and the delay to the resumption of menstruation is collected; and 3) the clinical study where an attempt is made to measure the delay to the resumption of ovulation directly. Each of these 3 types of studies is reviewed, underscoring why most quantitative assertions which derive from these studies about the magnitude of the relationship between nursing and its impact on fertility are conditional at best and in many respects still speculative. 1 promising line of research currently being pursued by Frisch (1975, 1978) is that the amount of reserve energy stored by the nursing mother in the form of fat is a critical intermediate variable in determining the demographic impact of lactation. Another promising line of research is concerned with the hormonal mechanisms involved in letdown. Until the individual data are sampled and aggregated to reflect conditions in the population at large, the kinds of exact inferences about the demographic impact of lactation will continue to be elusive.     

High-performance liquid chromatographic determination of bradykinin in saliva: a critical review and a new method

Because of difficulties or dubious results with previously published methodologies, a new semi-automated HPLC method with UV absorbance detection was developed and applied to the determination of bradykinin (BK) in human saliva. The new method consisted of an uncomplicated sample preparation involving the addition to saliva of an equal volume of 0.1 M orthophosphoric acid to stabilize BK, vortex-mixing, centrifugation, and separation, followed by chromatography of the supernatant phase on a C₈, 150×3.9-mm (I.D.) stainless steel column. The mobile phase was composed of 19% acetonitrile/0.1% trifluoroacetic acid at flow-rate of 0.4 ml/min. Using UV detection at 220 nm, the detection limit was 1 ng/ml for the BK standard, and 7 ng/ml for the assay of endogenous salivary BK. The orthophosphoric acid initially added to the saliva allowed BK to be stabilized from enzymic degradation at 20°C for 5 days and at 4°C for 60 days. Assignment made to the peak with the chromatographic properties of salivary BK was confirmed by HPLC–MS with an electrospray interface. This paper presents a new method that is reproducible, reliable and allows kinetic studies of salivary BK to be performed for clinical investigations.     

A critical review of mathematical models and data used in diabetology

The literature dealing with mathematical modelling for diabetes is abundant. During the last decades, a variety of models have been devoted to different aspects of diabetes, including glucose and insulin dynamics, management and complications prevention, cost and cost-effectiveness of strategies and epidemiology of diabetes in general. Several reviews are published regularly on mathematical models used for specific aspects of diabetes. In the present paper we propose a global overview of mathematical models dealing with many aspects of diabetes and using various tools. The review includes, side by side, models which are simple and/or comprehensive; deterministic and/or stochastic; continuous and/or discrete; using ordinary differential equations, partial differential equations, optimal control theory, integral equations, matrix analysis and computer algorithms.     

Pollution biomarkers in estuarine animals: critical review and new perspectives

In this review, recent developments in monitoring toxicological responses in estuarine animals are analyzed, considering the biomarker responses to different classes of pollutants. The estuarine environment imposes stressful conditions to the organisms that inhabit it, and this situation can alter their sensitivity to many pollutants. The specificity of some biomarkers like metallothionein tissue concentration is discussed in virtue of its dependence on salinity, which is highly variable in estuaries. Examples of cholinesterase activity measurements are also provided and criteria to select sensitive enzymes to detect pesticides and toxins are discussed. Regarding non-specific biomarkers, toxic responses in terms of antioxidant defenses and/or oxidative damage are also considered in this review, focusing on invertebrate species. In addition, the presence of an antioxidant gradient along the body of the estuarine polychaete Laeonereis acuta (Nereididae) and its relationship to different strategies, which deal with the generation of oxidative stress, is reviewed. Also, unusual antioxidant defenses against environmental pro-oxidants are discussed, including the mucus secreted by L. acuta. Disruption of osmoregulation by pollutants is of paramount importance in several estuarine species. In some cases such as in the estuarine crab Chasmagnathus granulatus, there is a trade off between bioavailability of toxicants (e.g. metals) and their interaction with key enzymes such as Na(+)-K(+)-ATPase and carbonic anhydrase. Thus, the metal effect on osmoregulation is also discussed in the present review. Finally, field case studies with fish species like the croaker Micropogonias furnieri (Scianidae) are used to illustrate the application of DNA damage and immunosuppressive responses as potential biomarkers of complex mixture of pollutants.     

Glass fibers covered with sol-gel glass as a new support for affinity chromatography columns: a review.

The search for mechanical supports for biochemically active compounds serving as immunochemical sensors has been the goal of many studies. A new compound in the form of gel fiberglass (GFG) membranes was recently developed as an example of such supports. In this review, these membranes were analyzed with respect to their use for cancer detection. The membranes are prepared from glass fibers covered with oxysilane to provide a sol-gel glass matrix. Derivatization of the support eliminates nonspecific adsorption. A thin layer of protein trapped in the gel glass during its preparation is deposited on the surface of a lattice of glass fibers. The major innovation of the membrane is its large area. External agents percolating through such a membrane contact a maximal number of molecules of the compounds trapped in the sol-gel glass. As a result, this membrane is highly effective. Each GFG column is built from a series of 20 to 30 membranes. The preparation of such columns is relatively simple, requiring only several hours. The capacity of GFG columns is high: the total amount of tumor-associated antigens (TAA) isolated by these columns from the blood of colon cancer patients reached 50% of the total protein and amounted to up to 9-12 mg/ml of serum. The main components of the isolated TAA were the soluble p66 and p51 proteins. A determination of their concentration by HPLC can be used for early cancer detection. Thus, the described method allows the easy and highly effective isolation of TAA and can be used for different goals, including cancer diagnosis. GFG supports have great potential for the isolation of various macromolecules utilizing specific ligands.     

A critical review of concepts and methods used in classical genome analysis

A short account of the development of classical genome analysis, the analysis of chromosome behaviour in metaphase I of meiosis, primarily in interspecific hybrids, is given. The application of the concept of homology to describe chromosome pairing between the respective chromosomes of a pair during meiosis is traced, and the relationship between this use of homology and the concept of homology as common ancestry is discussed at length. To equate the two concepts has led to the erroneous assumption that levels of chromosome pairing is an indication of phylogenetic relationship. Even accepting the fundamental premises, genome analysis is burdened by observational difficulties. Hence, chromosome pairing has been shown to be under genetic control and is also influenced by environmental conditions. Additionally, basic biological observations such as the distribution of meiotic configurations or the identity of the individual chromosomes are frequently neglected. Data from chromosome pairing are captured as pair-wise comparisons and are amenable only to phenetic analysis, and hence are not suited for phylogenetic inferences. As currently perceived, genome analysis may have a role to play in plant breeding but it has no place in systematics. With an increased knowledge and understanding of the mechanism behind meiosis, data useful in a systematic context may eventually be produced.     

Puberty as a critical risk period for eating disorders: A review of human and animal studies

This article is part of a Special Issue “Puberty and Adolescence”.     

Evaluation of new affinity chromatography resins for polyclonal,oligoclonal and monoclonal antibody pharmaceuticals

The performance of MabSelect SuRe and IgSelect affinity chromatography resins designed for process-scale purification of antibodies was investigated. Various antibodies (4 human monoclonal, 1 human polyclonal and 1 bovine polyclonal antibody and 1 Fc-fusion protein) were used to evaluate the elution pH and dynamic binding capacity of the resins. The elution pH for each human antibody was similar on MabSelect SuRe and IgSelect (pH 3.5–3.8). No significant differences in dynamic binding capacity were observed among human antibodies on MabSelect SuRe (∼20–40 mg/mL resin) and IgSelect (∼10–30 mg/mL resin). The binding capacity order for the human antibodies was the same on MabSelect SuRe and IgSelect. Using a linear pH gradient, both resins were able to partially separate monomeric and aggregated forms of the antibodies. The results indicate that these new affinity resins are powerful tools for the purification of human polyclonal antibodies from transgenic animals and oligoclonal antibodies from CHO cell cultures.     

Improved chromatographic separations on anion-exchange resins: I. Partition chromatography of sugars in ethanol

A resin conditioning/regeneration procedure is described which markedly enhances separation factors in partition chromatography of sugars on a commercially available anion-exchange resin. A mixture of 18 monosaccharides is well separated in less than 3 hr. A single ethanol-water solution is used as the eluent and no column temperature changes are necessary during a run. Sugars are located with a noncorrosive reagent.