[Ways to psychosomatic rehabilitation--differential treatment selection between psychosomatic acute care and rehabilitation]. / Wege in die psychosomatische Rehabilitation--differenzielle Indikationsstellung an der Schnittstelle zwischen Krankenhaus und psychosomatischer Rehabilitationsklinik.

Consecutive admissions to the psychosomatic departments of 5 hospital units in southwest Germany registered between October 2012 and October 2013 were asked to participate in a study investigating the treatment selection process for psychosomatic rehabilitation. 527 patients were included in the study, 269 outpatients and 258 inpatients at the end of their inpatient treatment. 52 patients (10.1%) received the recommendation for rehabilitation. 47 (90.4%) could be followed up 3 months later. 22 patients had applied for a rehabilitation treatment, 11 (50%) had obtained an approval for their rehabilitation, 5 had still no answer and for 6 patients the request was refused. 4 of the latter had objected their refusal and were still waiting for an answer. Only one patient was already admitted to a rehabilitation center. Possible reasons for the low permeability at the interface between hospital care and rehabilitation are discussed.

Modulations in martensitic Heusler alloys originate from nanotwin ordering.

Heusler alloys exhibiting magnetic and martensitic transitions enable applications like magnetocaloric refrigeration and actuation based on the magnetic shape memory effect. Their outstanding functional properties depend on low hysteresis losses and low actuation fields. These are only achieved if the atomic positions deviate from a tetragonal lattice by periodic displacements. The origin of the so-called modulated structures is the subject of much controversy: They are either explained by phonon softening or adaptive nanotwinning. Here we used large-scale density functional theory calculations on the Ni2MnGa prototype system to demonstrate interaction energy between twin boundaries. Minimizing the interaction energy resulted in the experimentally observed ordered modulations at the atomic scale, it explained that a/b twin boundaries are stacking faults at the mesoscale, and contributed to the macroscopic hysteresis losses. Furthermore, we found that phonon softening paves the transformation path towards the nanotwinned martensite state. This unified both opposing concepts to explain modulated martensite.

Forty-eight new cases with infertility due to balanced chromosomal rearrangements: detailed molecular cytogenetic analysis of the 90 involved breakpoints.

A molecular cytogenetic study was performed on 48 infertile patients who were identified as carriers of balanced translocations (40 cases), inversions (6 cases) or insertions (2 cases) by means of banding cytogenetics. Cases with a Robertsonian translocation or pericentric inversion 2 or 9 were not included. In summary, 100 break-events occurred in these patients, and 90 different chromosomal regions were involved. Thus, this study confirmed the presence of abnormal karyotypes in a subgroup of patients seeking infertility treatment. Breaks were demonstrated to appear preferentially in GTG-light bands in these patients. Furthermore, the observed breakpoints were associated with genomic regions prone to instability due to the presence of segmental duplications. Nonetheless, further detailed molecular analysis will be necessary in the future to characterize the mechanisms and genetic basis for this phenomenon.

Acceptor-specific glucuronyl transfer catalyzed by beta-glucuronidase.

Highly purified rat liver microsomal or lysosomal beta-glucuronidase (beta-D-glucuronide glucuronosohydrolase, EC 3.2.1.31) catalyzes the specific transfer of glucuronly residues from phenyl-beta-D-[U-14C]glucuronide to acceptor sugars. Specificity requirements of acceptor sugars are found to be: pyranose structure, 4C1-conformation and equatorial position of C2 and C3 hydroxyl groups or pyranose structure, 1C4-conformation and equatorial position of C3 and C4 hydroxyl groups. The acceptor capacities of 30 monosaccharides and glycosides including di- and tri- saccharides conform to this prinicple. The specificity of the beta-glucuronidase catalyzed glucuronyl transfer is proved by the exclusive formation of beta-glucuronly (1--3)glycosidic linkages. Glucuronly transfer rates increase with increasing donor substrate and increasing acceptor sugar concentration. In the presence of 1 M acceptor sugar the ratio of the transfer rate to the rate of enzymatic hydrolysis is about 2:1. An 'acceptor substrate binding site' on the surface of the beta-glucuronidase molecule which brings the C3 hydroxyl function of the acceptor sugar close enough to the C1 atom of the glucuronyl residue, is postulated.

Aggregation properties of beta-galactosidase of human urine and degradation of its natural substrates by a purified preparation of the enzyme.

Acid beta-D-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) was purified to near homogeneity from normal human urine by two affinity chromatography steps. On polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate the major protein band had an apparent molecular weight of 59000, thus being 5000 daltons smaller than the protein purified from human liver. Upon gel filtration on Sephadex G-150 column the purified enzyme had an apparent molecular weight of 70000 of pH 7.0. At pH 4.0 partial aggregation to a dimer of an apparent molecular weight of 150000 was found. Addition of 0.1 M galactose caused at pH 3.5, but not at pH 4.0 and 7.0, an increased formation of multimeric beta-galactosidase which eluted with the void volume of the column. Crude beta-galactosidase from human urine showed a higher aggregation tendency than the purified enzyme. None of the conditions produced an enzyme species of an apparent molecular weight of less than 40000. pH-activity profiles were measured against p-nitrophenyl-beta-D-galactoside, 3H-labelled GM1-ganglioside, [3H]keratan sulfate and the pentasaccharide O-beta-(1 leads to 4)-[6-3H]galactopyranosyl-O-beta-(1 leads to 2)-2-deoxy-2-acetamidoglycopyranosyl-O-alpha-(1 leads to 6)-mannopyranosyl-O-beta-(1 leads to 4)-mannopyranosyl-2-deoxy-2-acetamidoglucopyranoside. While p-nitrophenyl-beta-D-galactopyranoside and GM1-ganglioside were optimally hydrolyzed at pH 4.0, keratan sulfate and the pentasaccharide were optimally degraded at pH 4.3 and pH 5.0, respectively. With the chromogenic substrate and with GM1-ganglioside Km values of 0.33 mM were calculated. At pH 3.5 the hydrolysis of the synthetic substrate did not follow Michaelis-Menten kinetics. Two enzyme species appeared with Km values of 0.006 mM and 3.2 mM, respectively. The affinity of beta-galactosidase for [3H]keratan sulfate and the 3H-labelled pentasaccharide was at least one order of magnitude lower than for the amphiphilic substrates. Keratan sulfate and GM1-ganglioside did not act as competitive inhibitors of p-nitrophenyl-beta-galactosidase at the concentration tested. These findings could be explained by the existence of different binding sites for the substrates used.

Assay of mercaptopurine in plasma using paired-ion high-performance liquid chromatography.

A sensitive, quantitative, and specific high-performance liquid chromatographic method for mercaptopurine in plasma is described. The analysis, in which mercaptopurine and the internal standard, 6-methylthio-2-hydroxypurine, are chromatographed as ion-pairs with heptane-sulfonic acid, employs a simple and rapid sample preparation based on deproteination using 60% trichloroacetic acid. Quantitation of plasma samples to 0.2 microgram of mercaptopurine/ml is reported. The retention times of the major metabolites do not interfere.

Dye ingress in molars: furcation to chamber floor.

An in vitro dye leakage study was performed to determine the incidence of patent furcal accessory canals following exposure of the pulp chamber to 5.25% sodium hypochlorite. One hundred extracted molars were labeled, endodontically opened, and irrigated for 1 h at 5-min intervals. The external furcations were exposed to 0.5% basic fuchsin for 24 h. Patency was determined by dye presence on the chamber floor. Statistical analysis revealed that first and second molars, regardless of arch position or location, demonstrated a patent accessory canal at a rate of 57% +/- 19.6% in the furcal area. It was concluded that accessory canal exposure to 5.25% sodium hypochlorite in the furcal area of molars produced patency which was demonstrable via passive methods--no vacuum or injection pressures were utilized.

Nematode test to estimate the hazard potential of solved contaminations.

The acute toxicity of lindane, pentachlorophenol and a fluortenside was examined using a recently developed ecotoxicity test with the terrestrial nematode species Panagrellus redivivus. The exposure was performed in aqueous solution over an investigation period of 96 h. For lindane a time dependent decrease of the survival rate (LC50 after 96 h: 0.4 mg/l) was observed which indicates a high cuticular permeability of the insecticide. For pentachlorophenol the LC50 (96 h) was 13 mg/l, for the fluortenside a value of 110 mg/l was determined.

Determination of formetanate hydrochloride in selected fruits by coupled-column cation exchange liquid chromatography.

A strong cation exchange (SCX) liquid chromatographic (LC) method is described for determination of formetanate hydrochloride residue in pome, citrus, and stone fruits. A test portion of fruit, homogenized with the peel left on, was blended with acidified acetonitrile and filtered. A portion of extract was finely filtered, and a 500 microL aliquot (ca 0.2 g test sample equivalent) was loaded onto an SCX solid-phase extraction (SPE) LC column, which replaced the injection loop of the LC injection valve. Cations were selectively enriched; noncations were eluted by acetonitrile in a pre-separation cleanup. Turning the valve to the inject position coupled the SPE column to an SCX analytical column for separation and detection at 250 nm. The mobile phase was 0.4M pH 3.0 ammonium phosphate buffer-water-acetonitrile (50 + 25 + 25). Formetanate cation was quantitated by peak area and regression coefficients from a 5-point linear calibration covering a 100-fold range. Recovery of duplicate fortifications of apple, pear, orange, and peach averaged 89-99% at the respective U.S. tolerances of 3, 3, 4, or 5 ppm and averaged 93-99% at one-tenth of the respective tolerance level. Peel pigments or variable peel bulk of crop varieties tested, as well as other endogenous fruit material, contributed interference that was below the 0.02 ppm limit of detection. In a 1991 limited survey comprising 15 samples, none were found violative. Residues were found in 2 samples, but only 1 measurement was quantifiable, near the 0.06 ppm limit of quantitation.

Fam40b is required for lineage commitment of murine embryonic stem cells.

FAM40B (STRIP2) is a member of the striatin-interacting phosphatase and kinase (STRIPAK) complex that is involved in the regulation of various processes such as cell proliferation and differentiation. Its role for differentiation processes in embryonic stem cells (ESCs) is till now completely unknown. Short hairpin RNA (shRNA)-mediated silencing of Fam40b expression in ESCs and differentiating embryoid bodies (EBs) led to perturbed differentiation to embryonic germ layers and their derivatives including a complete abrogation of cardiomyogenesis. Pluripotency factors such as Nanog, Oct4 and Sox2 as well as epigenetic factors such as histone acetyltransferase type B (HAT1) and DNA (cytosine-5)-methyltransferase 3-ß (Dnmt3b) were highly upregulated in Fam40b knockdown EBs as compared with control and scrambled EBs. To examine the relevance of Fam40b for development in vivo, Fam40b was knocked down in developing zebrafish. Morpholino-mediated knockdown of Fam40b led to severe abnormalities of the cardiovascular system, including an impaired expression of ventricular myosin heavy chain (vmhc) and of cardiac myosin light chain 2 (cmlc2) in the heart. We identified the gene product of Fam40b in ESCs as a perinuclear and nucleolar protein with a molecular weight of 96 kDa. We conclude that the expression of Fam40b is essential for the lineage commitment of murine embryonic stem cells (mESCs) into differentiated somatic cells via mechanisms involving pluripotency and epigenetic networks.

The effects of xenobiotics on hepatic lipid and lipo-protein metabolism.

The liver occupies a central position in lipid and lipo-protein metabolism. Its function includes lipid and lipoprotein biosynthesis, assembly, packaging, transport, secretion, uptake and degradation of lipoproteins. In addition, enzymes synthesized and secreted by the liver into the blood stream or remaining bound to the endothelial cells in the capillaries, affect lipoprotein metabolism in the circulation. Xenobiotics may influence each of these steps. The mechanisms include more specific actions such as hormone or transmitter agonism and antagonism, membrane effects (stabilization or changes in trans-membrane gradients), influence on protein synthesis, influence on lipid metabolism by induction or inhibition of involved enzymes, or more general actions such as disturbances or damage of cellular membranes and cellular function. Some of these effects can easily be described as pharmacological actions, more or less independent of specific requirements in the chemical structure of the xenobiotics. Others are linked to specific chemical substituents such as carboxyl or alcoholic hydroxyl groups allowing the formation of lipid-xenobiotic-conjugates and/or the channeling of xenobiotics into lipid metabolism. This review will give a short overview of the mechanisms of xenobiotic-influenced hepatic lipid and lipoprotein metabolism.

Surrogate-assisted determination of 2,3,7,8-tetrachlorodibenzo-p-dioxin in fish by electron capture capillary gas chromatography.

Surrogate spiking the sample with 1000 parts per trillion (pptr) 1,3,7,8-tetrachlorodibenzo-p-dioxin (1378-TCDD) has doubled analytical throughput in determining toxic 2378-TCDD (analyte) at the low part-per-trillion level in fish, using multicolumn high resolution liquid chromatographic cleanup before quantitation by capillary gas chromatography with electron capture detection. The 1378- and 2378-TCDD were recovered equally and were well separated by the capillary column so that the earlier-eluting surrogate did not interfere with the quantitation of levels of analyte many-fold lower. Matrix interference contributed less than 1% bias in surrogate quantitation. Using surrogate recovery to correct for analyte losses during analysis, accuracy averaged (n = 7) 105% in determining 18 or 45 pptr 2378-TCDD added to fish without detectable bioincurred analyte. Analyses of selected fish with bioincurred 2378-TCDD gave results comparable to earlier work where recovery correction required a second analysis of sample fortified with analyte. With surrogate fortification, repeatability of determination (n = 3 or 4) improved markedly to less than 5% relative standard deviation at 37-46 pptr.

Purification and characterization of 3'-phosphoadenylylsulfate: N-desulfoheparan sulfate sulfotransferase from arterial tissue.

A 3'-phosphoadenylsulfate: N-desulfoheparan sulfate sulfotransferase (EC 2.8.2.12) was purified 450-fold from the microsomal fraction of calf arterial tissue and separated from 3'-phosphoadenylylsulfate:chondroitin sulfotransferase (EC 2.8.2.5) activity. The enzyme has optimal activity at neutral pH, requires divalent cations (Mn2+, Mg2+, Ca2+) for maximal activity and exhibits specificity towards N-desulfoheparan sulfate, N,O-desulfoheparan sulfate and oligosaccharides derived therefrom. N,O-desulfoheparan sulfate tetrasaccharides serve as acceptor substrates only if the nonreducing terminus is occupied by glucuronic acid (not iduronic acid). The N,O-desulfoheparan sulfate sulfotransferase transfers [35S]sulfate from 3'-phosphoadenylyl[35S]sulfate to the 2-amino groups and to the 6-hydroxy groups of glucosamine units of the acceptor substrates. The ratio of N/O-sulfation ranged between 3:1 and 2:1. O-[35S]Sulfated unsaturated disaccharides were obtained from enzymatically labelled [35S]N-desulfoheparan sulfate by heparitinase degradation and subsequent deamination. Evidence for the O-sulfation at C-6 of the glucosamine units was provided by isolation of anhydromannose [35S]monosulfate, which was formed from uronosylanhydromannose [35S]monosulfate by beta-glucuronidase treatment. An N-desulfo-N-[1-14C]lacetylheparan sulfate deacetylase activity was copurified with the N-desulfoheparan sulfate sulfotransferase.

Two N-acetylgalactosaminyltransferase are involved in the biosynthesis of chondroitin sulfate.

Two N-acetylgalactosaminyltransferases, designated I and II, have been purified from the microsomal fraction of calf arterial tissue and separated on Bio-Gel A. N-Acetylgalactosaminyltransferase I was purified 450-fold. It requires Mn2+ for maximal activity and transfers N-acetylgalactosamine residues from UDP-[1-3H]GalNAc in beta-glycosidic configuration to the non-reducing terminus of the acceptor substrates GlcA(beta 1-3)Gal(beta 1-3)Gal, GlcA(beta 1-3)Gal(beta 1-4)Glc and GlcA(beta 1-3)Gal. Even-numbered chondroitin oligosaccharides serve as acceptors for N-acetylgalactosaminyltransferase II, which transfers N-acetylgalactosamine from UDP-[1-3H]GalNAc to the non-reducing glucuronic acid residues of oligosaccharide acceptor substrates. Maximum transfer rates were obtained with a decasaccharide derived from chondroitin. Longer or shorter-chain chondroitin oligosaccharides are less effective acceptor substrates. All reaction products formed by N-acetylgalactosaminyltransferases I and II are substrates of beta-N-acetylhexosaminidase, which splits off the transferred [1-3H]GalNAc completely. In the microsomal fraction N-acetylgalactosaminyltransferase II had a 300-fold higher specific activity than N-acetylgalactosaminyltransferase I. In contrast to enzyme I, enzyme II loses much of its activity during the purification procedure and undergoes rapid thermodenaturation. GlcA-Gal-Gal is a characteristic sequence of the carbohydrate-protein linkage region of proteochondrioitin sulfate. The acceptor capacity of this trisaccharide suggests that N-acetylgalactosaminyltransferase I is involved in the synthesis of the carbohydrate-protein linkage region. Since N-acetylgalactosaminyltransferase II is highly specific for chondroitin oligosaccharides, we conclude that it participates in chain elongation during chondroitin sulfate synthesis.

Purification and characterization of a 3'-phosphoadenylylsulfate:chondroitin 6-sulfotransferase from arterial tissue.

A 3'-phosphoadenylylsulfate:chondroitin sulfotransferase (EC 2.8.2.5) was purified to homogeneity (about 760-fold) from the cytosolic fraction of calf arterial tissue by Con A-Sepharose, ion exchange and affinity chromatography. The enzyme has a molecular mass of 38000 Da, optimal activity at pH 6.0 (100%) and 7.25 (75%), requires divalent cations for maximal activity (Mn2+ greater than Mg2+, Ca2+) and exhibits specificity towards desulfated chondroitin sulfate and oligosaccharides derived therefrom. The enzyme transfers sulfate groups from [35S]phosphoadenylylsulfate exclusively to C-6 OH groups of N-acetylgalactosamine units of the acceptor substrates. Maximal sulfate transfer occurs at 2mM chondroitin disaccharide units (100%), the transfer rates decreasing with decreasing chain length in the order deca (55%), octa (17%) and hexasaccharides (4%). Lineweaver-Burk plots revealed equal maximal velocities for chondroitin, deca-, octa- and hexasaccharide, but decreasing Km values. Chondroitin 4-sulfate has 21% of the acceptor potency exhibited by chondroitin, whereas dermatan sulfate, heparan sulfate and hyaluronate and the chondroitin tetrasaccharide showed no acceptor properties. Analysis of the reaction products formed by prolonged enzymatic sulfation of a reduced chondroitin hexasaccharide [GlcA-GalNAc]2-GlcA-GalNAc-ol revealed that the preterminal N-acetylgalactosamine from the non-reducing end and the internal N-acetylgalactosamine but not the N-acetylgalactosaminitol were sulfated and that no hexasaccharide disulfate was formed by the action of chondroitin 6-sulfotransferase. Chondroitin 6-sulfotransferase is considered to possess a binding region capable of accommodating a nonsulfated oligosaccharide sequence of at least six sugars and is believed to act in the course of chondroitin sulfate synthesis in cooperation with, but shortly after, the enzymes involved in the chain elongation reaction.

Picogram level quantitation of 2,3,7,8-tetrachlorodibenzo-p-dioxin in fish extracts by capillary gas chromatography/matrix isolation/Fourier transform infrared spectrometry.

For the first time, gas chromatography/matrix isolation/Fourier transform infrared spectrometry (GC/MI/FTIR) has been reported to confirm the identity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2378-TCDD) and to quantify its level in fish extracts in the 170-220 pg range "on disk". When expressed on a fish tissue basis, analyte levels ranged from 15 to 45 pg/g. Spectroscopic identification was based on the position and relative intensity of seven absorption bands. Optical alignment as well as performance evaluation and optimization of the GC/MI/FTIR system are described. The use of [13C12]2378-TCDD as an internal standard was essential for quantitation, and quality assurance controls were used to verify system performance. GC/MI/FTIR quantitation of 2378-TCDD was compared with that independently found by GC with electron capture detection. Recovery of 2378-TCDD averaged 52% (n = 8, 30% relative standard deviation) for fish extracts.

Determination of 2,3,7,8-tetrachlorodibenzo-p-dioxin in fish, using electron capture gas chromatography with confirmation by mass spectrometry: interlaboratory study.

An interlaboratory study of the determination of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) in fish was conducted by 6 analysts in 4 laboratories using high resolution gas chromatography with electron capture detection (HRGC-EC) for quantitative screening analysis. Samples consisted of 3 Great Lakes channel catfish homogenates containing different levels of bioincurred 2,3,7,8-TCDD; 1 of these was prepared in duplicate and another was prepared both with and without standard 2,3,7,8-TCDD fortification for a total of 5 samples per set. All methods used included addition of 1,3,7,8-TCDD surrogate (to correct for procedural losses) followed by ethanolic KOH digestion and hexane extraction. Certain cleanup steps used, including sulfuric acid washing and multidimensional column liquid chromatographic procedures, varied among laboratories. Mean HRGC-EC results for the bioincurred residues were 56.6, 25.2, and 7.7 pg/g (ppt) with corresponding relative standard deviations (RSDs) of 9.1, 18.6, and 53.2%. Average determination of standard 2,3,7,8-TCDD from the fortified sample (corrected for surrogate recoveries averaging 74.6%) was 106% of the added amount (30.9 pg/g) with 11.0% RSD. HRGC-multiple ion detection mass spectrometry (MS), monitoring 12 ions, was used for confirmation. With the exception of several results from 1 analyst, HRGC-MS and HRGC-EC quantitations were in good agreement. All but 1 result reported met all of the MS identity criteria.