Characterization of DEAE-dextran by means of light scattering and combined size-exclusion chromatography/low-angle laser light scattering/viscometry

Ten DEAE (2-(diethylamino)ethyl) dextran samples were investigated by means of static and dynamic light scattering, viscometry and size-exclusion chromatography (SEC) in combination with on-line small-angle laser light scattering (LALLS) and viscometry (VISC). In dilute solution the behavior of DEAE-dextran was compared with that of unsubstituted dextran and the molecular weight M dependences of the radius of gyration R_g, hydrodynamic radius R_h, intrinsic viscosity [η], second virial coefficient A₂ and z-average diffusion coefficient D _z were determined. The relationships for DEAE-dextran dissolved in a 0,8 molar sodium nitrate solution were nearly the same as for dextran dissolved in water with 0,05 wt.-% sodium azide and gave the same exponents. The molecular weight dependence of the intrinsic viscosity cannot be described by a Kuhn-Mark-Houwink relationship with a constant exponent. The slope in the plot of log [η] versus log M decreases with increasing molecular weight which indicates the occurrence of branching. By means of SEC/LALLS/VISC measurements the molecular weight distributions were determined. The distributions were calculated (1) directly from the light scattering signal, (2) from a calibration line obtained by light scattering data of a DEAE-dextran sample with a broad distribution and (3) from the intrinsic viscosity distribution obtained by the on-line viscosity/refractive index detector in combination with the [η]-M relationship. In order to get the correct molecular-weight dependence of the intrinsic viscosity it is necessary to determine the molecular weight distribution directly by LALLS (technique 1) and to combine this with the appropriate intrinsic viscosity data from the viscometer. Only the third technique, which is an extension of technique 1, gave satisfactory results over the whole molecular weight region observed.     

Negative pressure effects during the isothermal crystallization of isotactic poly(propylene) studied by light and atomic force microscopy

The formation of holes during the late stage of the isothermal crystallization in thin films of isotactic poly(propylene) between two cover glasses was observed by light microscopy and atomic force microscopy. This behavior can be described consistently by the well-known negative pressure effect. Light microscopy reveals the simultaneous and sudden occurrence of a large number of small holes at the liquid-solid interface after the liquid in front of the spherulites is completely confined by other spherulites for a certain time interval. In exceptional cases only a few holes appear and finally large cavities are formed. Atomic force microscopy measurements carried out in the height mode are able to prove the hole formation in front of the spherulites. Furthermore, a substantial thinning of the two-dimensional spherulites in thin films can be observed prior to the hole formation.     

Plasma cystine concentration and redox state in aging and physical exercise

Because redox-regulated signalling pathways are often modulated by the thiol/disulfide redox state (REDST), changes in plasma REDST may possibly account for pathological processes. We, therefore, investigated the mechanisms that account for changes in the plasma REDST as derived in first approximation from the cystine and acid soluble thiol (mainly cysteine) concentrations. Elderly subjects (studies A) and younger subjects after intensive physical exercise (IPE) (study B) i.e. subjects in conditions typically associated with decreased insulin responsiveness, showed, on the average, an increase in the plasma total free amino acid (TAA) concentration to approximately 3000 microM, including an increase in cystine but no increase in the thiol concentration if compared with controls. The REDST was decreased accordingly. A study on the postabsorptive amino acid exchange rates across the lower extremities (study C) indicated that a TAA level > or =2800 microM supports a balanced net protein synthesis even under conditions of weak insulin stimulation, suggesting that high TAA levels may prevent the release of cysteine into the blood in the postabsorptive state. Collectively, these studies indicate that the age-related oxidative shift in plasma REDST may result from the decrease in amino acid clearance capacity and may be aggravated by excessive physical exercise.     

Temperature rising elution fractionation of a random ethene/styrene copolymer

A random ethene/styrene copolymer containing 13.8 mol-% styrene was prepared with the Ziegler-Natta catalyst system Me₂Si(Me₄Cp)(N-t-butyl)TiCl₂/methylaluminoxane and characterized by means of preparative temperature rising elution fractionation (TREF) combined with size exclusion chromatography, NMR, differential scanning calorimetry and wide-angle X-ray scattering analyses of the copolymer fractions. Efforts are made to describe the distribution of the styrene content of the copolymers using the Stockmayer-Tacx distribution function. Both, comonomer distribution and molar mass distribution strongly support the presence of a single type of catalytically active center.     

Knowledge representation model for systems-level analysis of signal transduction networks

A Petri-net based model for knowledge representation has been developed to describe as explicitly and formally as possible the molecular mechanisms of cell signaling and their pathological implications. A conceptual framework has been established for reconstructing and analyzing signal transduction networks on the basis of the formal representation. Such a conceptual framework renders it possible to qualitatively understand the cell signaling behavior at systems-level. The mechanisms of the complex signaling network are explored by applying the established framework to the signal transduction induced by potent proinflammatory cytokines, IL-1beta and TNF-alpha The corresponding expert-knowledge network is constructed to evaluate its mechanisms in detail. This strategy should be useful in drug target discovery and its validation.     

Characterization of monocyte-derived dendritic cells in recent-onset diabetes mellitus type 1

Dendritic cells (DC) may play an important role in the immunopathogenesis of type 1 diabetes mellitus (DM-1). In this study, we have analyzed phenotypical changes during cytokine-driven maturation from CD14+ monocytes to mature DC and DC-dependent T-cell stimulation in recent-onset pediatric DM-1 patients and healthy controls. DC maturation was monitored by flow cytometric analyses for the expression of surface markers (HLA-DR, CD1a, CD40, CD80, CD86, CD83, CD14, CD32, mannose-receptor, and CD11c). Flow cytometric analysis of isolated peripheral blood monocytes did not reveal apparent differences between patients and controls. During DC maturation no obvious differences in the expression patterns of surface markers over time or evidence for maturation impairments in DM-1 patients could be appreciated. Solely, a marginal, but significant, transient down-regulation of CD1a on Day 3 (mean MDFI 3.82 vs 7.25; P = 0.021), which was accompanied by an increase of IL-6, could be observed. The comparison of mature DCs (Day 10) between patients and controls indicated no significant differences, except for CD83 (mean MDFI 1.7 vs 1.5; P = 0.042) and CD80 (mean MDFI 15.92 vs 12.73; P = 0.042). Moreover, no difference in T-cell stimulatory capacity was seen. In conclusion, our analysis of a cohort of recent-onset DM-1 patients and controls does not support a role for disease-related alterations in cytokine-driven maturation of monocyte-derived DC.     

Microproteins in amniotic fluid as an index of changes in fetal renal function during development

Protein content and protein composition were studied in amniotic fluid obtained from 171 healthy pregnant women between the 16th and 38th week of gestation, using microgradient gel electrophoresis to separate proteins according to their molecular size into albumin (68 KD), proteins of low molecular weight (LMW proteins, <68 KD), and proteins of high molecular weight (HMW proteins, >68 KD). Additionally -1-microglobulin (-1-MG, 33 KD) and -2-microglobulin (-2-MG, 11,8 KD) were analysed as micromolecular marker proteins. Concentrations of LMW proteins were 0.15–0.22 g/l, of -1-MG 28.4–34.5 mg/l, and of -2-MG 7.2–11.6 mg/l during the second trimester of gestation, and thereafter decreased progressively to 0.03 g/l, 14.1 mg/l and 2.4 mg/l respectively near term. The same developmental trends were confirmed by calculating the protein/creatinine ratios in amniotic fluid. The concentrations of LMW proteins found in the first postnatal urine of 73 healthy infants born prematurely or at term were similar to those in amniotic fluid of corresponding fetal age. Concentrations of albumin and HMW proteins in postnatal urine were about 5% and 15% respectively when compared with amniotic fluid concentrations. No strong correlation existed between gestational age and either of the analysed proteins which would allow accurate assessment of fetal maturation by protein analysis in amniotic fluid. It is concluded that fetal urinary excretion is the major determinant of the microprotein content of amniotic fluid. Microproteins seem to reflect an increasing tubular reabsorption capacity, which accelerates rapidly after the second trimester of gestation.     

Using Longitudinal Social Network Analysis to Evaluate a Community-Wide Parenting Intervention

Prevention Science - Harsh parenting attitudes and behaviors negatively impact children’s behavior and development, and are linked to heightened levels of violence in children. Parent...     

Diagnostic value of routine semen analysis in clinical andrology

Infertility is a major health issue affecting over 48.5 million couples around the world, with the male factor accounting for about 50% of the cases. The conventional semen analysis recommended by the World Health Organization (WHO) is the cornerstone in the evaluation of male fertility status. It includes macroscopic and microscopic evaluation of the ejaculate, which reflects the production of spermatozoa in the testes, the patency of the duct system and the glandular secretory activity. Evaluation of seminal fructose, sperm vitality and leucocytes (Endtz test) are useful adjuncts to semen analysis that provide information on specific clinical conditions. Though several computer-assisted sperm analysis (CASA) systems have been developed, conventional methods for semen analysis are still widely accepted in clinical practice. This review summarises the conventional techniques used in routine semen analysis and their diagnostic value in clinical andrology.     

Protein profiling in unlocking the basis of varicocele-associated infertility

Varicocele is one of the major causes of male infertility and has a negative impact on spermatogenesis. The conventional semen analysis does not reveal the underlying subcellular mechanisms associated with defects in spermatozoa. Proteomics and bioinformatics analysis can be used to identify the molecular aetiologies associated with poor semen quality in varicocele patients. Mitochondrial dysfunction has been identified as the main factor affecting normal physiological functions of spermatozoa. This article discusses the proteomic studies of spermatozoa and seminal plasma in varicocele patients. Proteomics can identify potential spermatozoa and seminal plasma biomarkers in varicocele-mediated male infertility. In future, these protein biomarkers can be useful in the development of noninvasive diagnostic and therapeutic strategies for varicocele patients.