A strategy for the comparative analysis of serum proteomes for the discovery of biomarkers for hepatocellular carcinoma

Many of the emerging technologies for the global evaluation of gene expression, at both the RNA and protein level, are being applied to the problem of finding biomarkers for human disease progression. These analyses can be made difficult, however, by variation between samples that arises from both technical and nondisease related physiological or genetic causes. In an effort to identify serum polypeptides whose presence or absence correlates with the clinical status of patients at high risk for hepatocellular carcinoma (HCC), we have developed a strategy that helps to focus the analysis on meaningful changes in protein levels above the background of variation. For the current study we divided the patient population into four clinically defined diagnostic groups that represent a generally increasing risk for HCC. Chronic infection with hepatitis B virus (HBV) is a major risk factor for HCC and our groups included patients with no indication of liver disease (healthy), those with inactive chronic HBV, those with active chronic HBV, and patients with a diagnosis of HCC and history of chronic HBV infection. Serum polypeptides from these patients were first analyzed in two-dimensional gels by combining the serum from patients in each of the four groups to generate composite gel profiles. Analysis of these composite gels allowed us to identify two relatively abundant features that were reduced in the HCC group as compared to the healthy group. Tryptic fragment mass fingerprinting identified the features as a carboxy terminal fragment of complement C3 and an isoform of apolipoprotein A1. These two features were examined by two-dimensional gel electrophoresis of serum from each individual in the four groups in order to verify that the inter-group differences seen in composite gels reported changes in abundance for most members of the group, rather than extreme changes for a small fraction of the group. These preliminary studies suggest that a proteomic methodology can be used for the identification of serum biomarkers for HCC and other liver disease.     

A Procedure for the Design of Novel Assisting Devices for the Sit-to-Stand

The Sit-to-Stand （STS） is an activity most people perform numerous times daily. Standing up deals with the transition from two stabilized postures, namely seated to standing, with movement of all body segments except the feet. During the STS the body＇s Center of Gravity （COG） is moved upward from a sitting position to a standing position without losing balance and requiring a good coordination of many muscles. Three main phases of the STS movement can be recognized. One begins to stand up by inclining the upper body forward, which moves body mass toward the feet in order to maintain balance after lift-off. Prior to leaving the chair, hip and knee extensor muscles are activated to provide antigravity support for these joints, this action is commonly referred to as ＂weight shift＂. Finally; after leaving the chair, the leg and trunk joints are straightened to achieve upright stance. The STS task can be considered of major importance for impaired and elderly people to achieve minimal mo- bility and independence. In this paper we detail a procedure for the design of assisting devices to be used for the STS. In par- ticular, an experimental procedure is described firstly to track and record point trajectories and the orientation of the trunk during the STS. This analysis is then used to get information for the design of assisting devices. A proposal and simulation results are presented for a novel mechatronic system. In particular, for the case under study experimental tests are used to drive the actua- tion system for the reported simulation. A functional mechatronic scheme is then proposed to control the device during its operation.     

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.     

Template for developing guidelines for the evaluation of the clinical efficacy of psychophysiological interventions

An essential function of both the Association for Applied Psychophysiology and Biofeedback (AAPB) and the Society for Neuronal Regulation (SNR) is the systematic evaluation of psychophysiological interventions that have been developed for the treatment of medical and psychiatric disorders. In order to address scientific concerns regarding the efficacy of specific clinical applications of biofeedback, these two societies formed and Efficacy Task Force. The process to be used in the assessment of treatment efficacy, specificity and clinical utility is presented in the form of a template that will serve as the foundation for a series of scientific reviews and practice guidlines to be published by both societies.     

A model for the functioning of the striatum

A model is presented for the operation of the striatum. The model posits that the basal ganglia are responsible for driving smooth transitions of state for an organism. We propose that this is accomplished through the computation of a potential function within the striatum on which a gradient descent is performed toward the goal state. The model suggests that various somatotopic regions of the striatum correspond to state spaces, each of which pertains to a different aspect of the organism. This paper discusses this model only in the context of motor control, i.e., egomotion and limb movement. The model appears to account for a variety of experimental results, and for some unusual properties of the striatum.     

Structural basis for the lower affinity of the insulin-like growth factors for the insulin receptor

Insulin and the insulin-like growth factors (IGFs) bind with high affinity to their cognate receptor and with lower affinity to the noncognate receptor. The major structural difference between insulin and the IGFs is that the IGFs are single chain polypeptides containing A-, B-, C-, and D-domains, whereas the insulin molecule contains separate A- and B-chains. The C-domain of IGF-I is critical for high affinity binding to the insulin-like growth factor I receptor, and lack of a C-domain largely explains the low affinity of insulin for the insulin-like growth factor I receptor. It is less clear why the IGFs have lower affinity for the insulin receptor. In this study, 24 insulin analogues and four IGF analogues were expressed and analyzed to explore the role of amino acid differences in the A- and B-domains between insulin and the IGFs in binding affinity for the insulin receptor. Using the information obtained from single substituted analogues, four multiple substituted analogues were produced. A "quadruple insulin" analogue ([Phe(A8), Ser(A10), Thr(B5), Gln(B16)]Ins) showed affinity as IGF-I for the insulin receptor, and a "sextuple insulin" analogue ([Phe(A8), Ser(A10), Thr(A18), Thr(B5), Thr(B14), Gln(B16)]Ins) showed an affinity close to that of IGF-II for the insulin receptor, whereas a "quadruple IGF-I" analogue ([His(4), Tyr(15), Thr(49), Ile(51)]IGF-I) and a "sextuple IGF-II" analogue ([His(7), Ala(16), Tyr(18), Thr(48), Ile(50), Asn(58)]IGF-II) showed affinities similar to that of insulin for the insulin receptor. The mitogenic potency of these analogues correlated well with the binding properties. Thus, a small number of A- and B-domain substitutions that map to the IGF surface equivalent to the classical binding surface of insulin weaken two hotspots that bind to the insulin receptor site 1.     

Aspects for the future of biotransformations

Biotransformations have gained extensive importance in practical use as a support for chemical synthesis or in the conversion of natural products. Biotransformations may present an enlargement, a sequential degradation or a specific modification of synthetic or natural compounds. The tools for biotransformations are principally mammalian, plant or microbial cells and their cell-free enzymes. In technical practice the biocatalysts are so far limited to the use of microorganisms and some cell-free enzymes of low cost. Although numerous microbial or enzymatical reactions were already developed for industrial processes, the capacities of biotransformations offer a broad field of inexhaustible possibilities for the future.

     

On the nature of mechanisms responsible for the higher efficiency of urea supergranules for rice

Summary A laboratory study made with a sandy clay loam soil of pH 7.7 showed that hydrolysis of urea was slower when applied as urea supergranules (USG) than commercial urea prills. This resulted in maintenance of applied nitrogen as ammonium in the soil for a longer period. Nitrite concentration was also much less with USG. These three mechanisms are responsible for the higher efficiency of USG over urea prills.     

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.     

Criteria for the selection of the most desirable radionuclide for radiolabeling monoclonal antibodies

Efficient labeling of monoclonal antibodies depends on a number of key factors, mostly related to the characteristics of the radionuclide itself and to the manner of its incorporation into the protein. Such factors include the physical half-life, the photon or particle energy of the radionuclide and its selective deposition of energy in tissues, the method of labeling used (covalent binding or chelation), and the effect that the chemical changes inherent in the labeling process may have on the properties of the protein or of its fragments. The major biological factor in determining the radionuclide of choice for labeling is the projected use of the labeled antibody. When the intended use is diagnostic, then what is required is high-photon density for achieving the high resolution needed for imaging, whereas therapeutic use requires radionuclides with high energy deposition at the target sites, i.e. β or α emitters. A further consideration is to be given to the mode of administration of the radiolabeled monoclonal antibody: determination of the radiopharmacokinetic parameters of compartmental models of biodistribution of the labeled monoclonal antibody and/or its fragments may also assist in selecting which radionuclide may be best to use for radiolabeling a given monoclonal antibody intended for either tumor diagnosis, prognosis and/or therapy.     

Program for the application of genetic transformation for crop improvement in the semi-arid tropics

Summary The semi-arid tropics are characterized by unpredictable weather, limited and erratic rainfall and nutrient-poor soils and suffer from a host of agricultural constraints Several diseases, insect pests and drought affect crop productivity. Developing stress-resistant crops has been a worthwhile activity of the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT). Mandated crops of ICRISAT, including groundnut, pigeonpea, chickpea, sorghum and pearl millet, are the main staple foods for nearly one billion people in the semi-arid tropics. Genetic transformation provides a complementary means for the genetic betterment of the genome of these crops. Judicious application of biotechnological tools holds great potential for alleviating some of the major constraints to productivity of these crops in the agricultural systems of the semi arid tropics. This article reviews plant genetic engineering in relation to its applications in genetic enhancement and the improvement of important crops of the semi-arid tropics. For the benefit of nonbiotechnologists, a brief review of technical aspects of plant genetic engineering is also included.     

An enzymatic pathway for the biosynthesis of the formylhydroxyornithine required for rhodochelin iron coordination

Rhodochelin, a mixed catecholate-hydroxamate type siderophore isolated from Rhodococcus jostii RHA1, holds two L-δ-N-formyl-δ-N-hydroxyornithine (L-fhOrn) moieties essential for proper iron coordination. Previously, bioinformatic and genetic analysis proposed rmo and rft as the genes required for the tailoring of the L-ornithine (L-Orn) precursor [Bosello, M. (2011) J. Am. Chem. Soc.133, 4587-4595]. In order to investigate if both Rmo and Rft constitute a pathway for L-fhOrn biosynthesis, the enzymes were heterologously produced and assayed in vitro. In the presence of molecular oxygen, NADPH and FAD, Rmo monooxygenase was able to convert L-Orn into L-δ-N-hydroxyornithine (L-hOrn). As confirmed in a coupled reaction assay, this hydroxylated intermediate serves as a substrate for the subsequent N(10)-formyl-tetrahydrofolate-dependent (N(10)-fH(4)F) Rtf-catalyzed formylation reaction, establishing a route for the L-fhOrn biosynthesis, prior to its incorporation by the NRPS assembly line. It is of particular interest that a major improvement to this study has been reached with the use of an alternative approach to the chemoenzymatic FolD-dependent N(10)-fH(4)F conversion, also rescuing the previously inactive CchA, the Rft-homologue in coelichelin assembly line [Buchenau, B. (2004) Arch. Microbiol.182, 313-325; Pohlmann, V. (2008) Org. Biomol. Chem.6, 1843-1848].     

A salt-concentration gradient method for the determination of the maximum salt concentration for microbial growth

A method is described for establishing a discontinuous salt concentration gradient which is useful for determining the maximum salt concentration which can be tolerated by growing microorganisms. The technique can be used aerobically or anaerobically, and has a standard error varying from ± 0.1% to ± 0.4%, depending on the organism. Data are given for several yeasts and forSerratia marcescens. The results indicate that the method has a potential usefulness for separating and identifying closely related microorganisms.     

Possibilities for a new breed of cattle for the south

Ohne Zusammenfassung

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Possibilities for a new breed of cattle for the south

     

检验医学专业临床实习管理存在的问题及对策

临床实习是医学理论和临床实践工作有机结合的连接点,是医学生走向临床工作必不可少的重要环节。临床实习教学是医学生教育中一个重要的组成部分,是从课堂到临床、从理论到实践、从理性到感性的重要过程,也是巩固基础知识、理论联系实际、走向独立工作岗位的重要一步。在临床实践过程中,教职人员要培养学生获取、分析和处理疾病信息的能力,正确书写病历、熟练掌握诊疗技术以及与患者及家属和谐沟通的能力。医学检验学是运用现代物理化学方法、手段进行医学诊断的一门学科,通俗的说,就是利用仪器、试剂、CT扫描等检测人的各项指标是否正常,为临床诊断提供依据。因此,加强临床检验医学专业的实习管理工作至关重要。本文从建立合理的教学管理体系、加强临床实习考核制度、完善实习管理、加强教师队伍建设等几个方面,探讨临床检验学实习管理存在的问题,并提出相应的对策,为高等医学院校教学管理工作提供思路。     

The role of mutants in the search for the photoreceptor for phototropism in higher plants

Early attempts to identify the chromophore of the photoreceptor for phototropism are reviewed. Carotenoids and flavins were the principal candidates, but studies with grass coleoptiles devoid of carotenoids suggest that at least in these organs carotenoids are most unlikely to play that role. The status of characterization of a gene for a putative photoreceptor protein is also reviewed. As the action spectrum for phototropism resembles the absorption spectrum of a flavoprotein, flavoproteins are attractive candidates at present, especially since the CRY1 photoreceptor in Arabidopsis thaliana that mediates blue light-dependent hypocotyl growth suppression has flavin adenine dinucleotide as one of its two chromophores. As the second chromophore appears to be pterin, pterins should not be ruled out as candidate chromophores for the photoreceptor for phototropism.     

Technical Procedures for the Study of the Nucleolonema

In a recent paper¹ the authors have shown filamentous structures within the nucleolus of all cells. This structure was named the nucleolonema. In the present paper the technical procedures which prove their existence are described in detail.

There are two groups: a) Silver impregnation procedures and iron-pyrogallic stain for fixed material; b) phase contrast microscopy, dark-field illumination and oblique transillumination for fresh material.

The suitable objects are mentioned in every case in order to obtain the best results.     

A method for assessing the efficacy of using bifidumbacterin for the prevention of suppurative-inflammatory diseases in the newborn

A method has been developed for the evaluation of the effectiveness of bifidumbacterin in different quantitative morbidity characteristics in purulent inflammatory diseases of newborns in risk groups. This method requires a limited number of observations. In purulent inflammatory infections bifidumbacterin can be used as an effective remedy for the prophylaxis of hospital infections. The proposed method may be used for the analysis of the effectiveness of other antiepidemic measures, e.g. the sanitation of carriers.     

The outlook for the development of live vaccines for the prevention of melioidosis

The effectiveness of immunization with Burkholderia pseudomallei attenuated strains (Pur and Ts), heterologous vaccines and the recombinant culture of Francisella tularensis RM2 carrying a plasmid with fragments of B. pseudomallei chromosome was studied in four species of experimental animals, essentially differing in their sensitivity to melioidosis. The most immunogenic B. pseudomallei mutants, introduced subcutaneously, created a statistically significant level of protection in animals, moderately sensitive to melioidosis, but proved to be ineffective in highly sensitive animal models when tested under the same conditions. In aerogenic infection the effectiveness of the experimental vaccines under study in all species of the animals was on the same level. The study showed good prospects of using tularemia vaccine for inducing heterologous immunity to melioidosis, as well as the possibility of its use as the basis of a bivalent gene-engineering vaccine.