[Structure of pain management facilities in Germany : Classification of medical and psychological pain treatment services-Consensus of the Joint Commission of the Professional Societies and Organizations for Quality in Pain Medicine]. / Struktur der schmerzmedizinischen Versorgung in Deutschland : Klassifikation schmerzmedizinischer Einrichtungen - Konsens der Gemeinsamen Kommission der Fachgesellschaften und Verbände für Qualität in der Schmerzmedizin.

On behalf of the Medical/Psychological Pain Associations, Pain Patients Alliance and the Professional Association of Pain Physicians and Psychologists, the Joint Commission of Professional Societies and Organizations for Quality in Pain Medicine, working in close collaboration with the respective presidents, has developed verifiable structural and process-related criteria for the classification of medical and psychological pain treatment facilities in Germany. Based on the established system of graded care in Germany and on existing qualifications, these criteria also argue for the introduction of a basic qualification in pain medicine. In addition to the first-ever comprehensive description of psychological pain facilities, the criteria presented can be used to classify five different levels of pain facilities, from basic pain management facilities, to specialized institutions, to the Centre for Interdisciplinary Pain Medicine. The recommendations offer binding and verifiable criteria for quality assurance in pain medicine and improved pain treatment.

Comparative clinical effects of hydromorphone and morphine: a meta-analysis.

We have conducted a meta-analysis of the clinical effects of morphine and hydromorphone to compare their benefit in analgesia. Embase and Medline were searched with an end-date of June 2009 for randomized, controlled trials or observational studies that addressed comparative analgesic and side-effects or particular side-effects. Two researchers independently identified included studies and extracted the data. Estimates of opioid effects were combined by using a random-effects model. Meta-analysis of eight studies suggested that hydromorphone (494 patients) provides slightly better (P=0.012) clinical analgesia than morphine (510 patients). The effect-size was small (Cohen's d=0.266) and disappeared when one study was removed, although the advantage of hydromorphone was more evident in studies of better quality (Jadad's rating). Side-effects were similar, for example, nausea (P=0.383, nine studies, 456 patients receiving hydromorphone and 460 morphine); vomiting (P=0.306, six studies, 246 patients receiving hydromorphone and 239 morphine); or itching (P=0.249, eight studies, 405 patients receiving hydromorphone, 410 morphine). This suggests some advantage of hydromorphone over morphine for analgesia. Additional potential clinical pharmacological advantages with regard to side-effects, such as safety in renal failure or during acute analgesia titration, are based on limited evidence and require substantiation by further studies.

[Recommendations on classification of German pain treatment services]. / Empfehlung zur Klassifikation Schmerztherapeutischer Einrichtungen in Deutschland.

On behalf of the German chapter of the International Association for the Study of Pain (IASP) recommendations for German pain treatment services have been developed for the first time. The criteria were based on the IASP recommendations but adapted to the specific German situation. According to the structure and process criteria four different levels of pain treatment services can be distinguished. The aim of the recommendations is to serve as a guide for future development and implementation of pain therapy and quality assurance.

Isolation of mitochondrial porin of the fly Protophormia: porin modification by the pesticide CGA 140'408 studied in lipid bilayer membranes.

Mitochondrial porin from the fly Protophormia was solubilized with detergent from whole mitochondria and purified by chromatography across a hydroxyapatite (HPT) column. The purified protein had an apparent molecular mass of about 30 kDa on SDS-PAGE. Partial sequencing of the protein confirmed that it is porin. When reconstituted in planar lipid bilayer membranes, porin formed ion-permeable channels with single-channel conductances of 2.4 and 4.5 nS in 1 M KCl. At low voltage, Protophormia porin displayed the properties of a general diffusion pore and had a small selectivity for anions over cations. At transmembrane potentials starting with about 20-30 mV, the channel switched in closed state, which is still ion-permeable. Our results suggest that Protophormia porin possesses functional properties similar to those of other mitochondrial porins. Porin was also isolated and purified from mitochondria, which were treated with the carbodiimide CGA 140'408 It represents the active derivative of diafenthiuron a new acaricide and insecticide. This carbodiimide labels both a F0-component of the inner membrane ATPase and outer membrane porin in a similar way as N,N'-dicyclohexylcarbodiimide (DCCD). Reconstitution experiments with the CGA 140'408-modified porin showed no significant effect of the modification on the single-channel conductance, suggesting that CGA 140'408 binds outside the channel. The voltage-dependence of the CGA 140'408-modified porin was changed with respect to the unmodified form. The closed configuration of the pesticide-modified channel was reached at smaller transmembrane potentials, suggesting a shift of the open to the closed state of Protophormia porin by pesticide binding. A possible contribution of this effect to the pesticide action is discussed.

Crystallization, crystal structure analysis and preliminary molecular model of the bilin binding protein from the insect Pieris brassicae.

The bilin binding protein of the butterfly Pieris brassicae has been prepared, crystallized and its crystal structure determined at high resolution using film and FAST area detector intensity data. The crystallographic asymmetric unit contains a tetramer of identical subunits with a molecular weight of about 90,000. The crystal structure was determined by isomorphous replacement. Use was made of the molecular symmetry to improve phases. A molecular interpretation of the electron density distribution and partial tracing of the polypeptide chain was possible without amino acid sequence information, as the fold is very similar to retinol binding protein. It is characterized by a beta-barrel formed by two orthogonal beta-sheets and an alpha-helix. The bilin pigment seems to be bound within the beta-barrel analogously to retinol in retinol binding protein. The tetramer in the crystal has C2 symmetry and is a dimer of dimers of quasi-equivalent subunits.

Molecular structure of the bilin binding protein (BBP) from Pieris brassicae after refinement at 2.0 A resolution.

The bilin binding protein (BBP) from the insect Pieris brassicae has been analysed for amino acid sequence, spectral properties and three-dimensional structure. The crystal structure that had been determined by isomorphous replacement has been refined at 2.0 A (1 A = 0.1 nm) resolution to an R-value of 0.20. The asymmetric unit contains four independent subunits of BBP. The co-ordinate differences are 0.25 A, in accord with the estimated error in co-ordinates. The polypeptide chain fold is characterized by an eight-stranded barrel. The connecting loops splay out at the upper end of the barrel and open it, whilst the lower end is closed. The overall shape resembles a calyx. The biliverdin IX gamma chromophore is located in a central cleft at the upper end of the barrel. The bilatriene moiety is in cyclic helical geometry with configuration Z,Z,Z and conformation syn,syn,syn. The geometry is in accord with the spectral properties and permits a correlation between sign of the circular dichroism bands and sense of the bilatriene helices. The fold of BBP is related to retinol binding protein (RBP), as had been recognized in the preliminary analysis, although the amino acid sequences of RBP and BBP show only 10% homology. There are large differences in the loops at the upper end of the barrel, whilst the segments of the centre and the lower end of the barrel superimpose closely. The ligands of BBP and RBP, biliverdin and retinol, respectively, are also similarly located.

Metabolic clearance rates of catechol estrogens in rats.

MCRs of the catechol estrogens 4-hydroxyestradiol (4-OHE2) and 2-hydroxyestradiol (2-OHE2) and of the parent estrogen 17 beta-estradiol (E2) were determined in rats. Long term ovariectomized Wistar rats were infused with the steroids at a constant rate for 3 days via a catheter placed in the abdominal aorta. Blood samples were drawn discontinually by retroorbital puncture, and the serum concentrations of E2, 4-OHE2, and 2-OHE2 were measured by RIA. Steady state was reached within 24 h of infusion. Mean serum MCRs were calculated to be 740 +/- 117 ml/h for E2, 2700 +/- 1000 ml/h for 4-OHE2, and 8300 +/- 1700 ml/h for 2-OHE2. Thus, the MCRs of the catechol estrogens were definitely higher than the MCR of E2 resulting in an apparent ratio of 1:4:11 (E2:4-OHE2:2-OHE2).

Stimulation of human peripheral blood lymphocytes by bioactive peptides derived from bovine milk proteins.

The in vitro modulation of the proliferation of human peripheral blood lymphocytes by different synthetic peptides derived from milk proteins was investigated. Therefore, proliferation changes were followed up after incorporation of BrdU into the DNA, and the influence on protein biosynthesis was measured using the [3H]leucine incorporation test. Tyr-Gly and Tyr-Gly-Gly significantly enhanced (maximal 90 and 35%, respectively) the proliferation of PBL. For beta-casomorphin-7 and beta-casomorphin-10,lymphocyte proliferation was suppressed at lower concentrations, but stimulated at higher concentrations (> or = 10(-7) mol/l). Protein synthesis was stimulated (maxima at 25%) only with Tyr-Gly and Tyr-Gly-Gly. The findings point to a need for further studies on the possible function of peptides derived from milk proteins as orally bioavailable immunopotentiatory compounds.

Incorporation of N-acyl-2-amino-2-deoxy-hexoses into glycosphingolipids of the pheochromocytoma cell line PC 12.

The synthesis of N-acyl-2-amino-2-deoxy-hexoses, their metabolism and their incorporation into glycosphingolipids of rat pheochromocytoma cell line PC 12 were investigated. The data indicate that in PC 12 cells the N-acyl-2-amino-2-deoxy-hexoses, N-propanoyl-D-glucosamine and N-butanoyl-D-glucosamine are metabolized to the corresponding phosphates, and that N-propanoyl-D-glucosamine is also metabolized to N-propanoyl neuraminic acid. Using variously radiolabelled N-acyl-2-amino-2-deoxy-hexoses, their incorporation into glycosphingolipids was shown.

5-Azidoimidacloprid and an acyclic analogue as candidate photoaffinity probes for mammalian and insect nicotinic acetylcholine receptors.

The 5-azido analogue of the major insecticide imidacloprid, 1-(5-azido-6-chloropyridin-3-ylmethyl)-2-nitroiminoimidaz olidine (1), and an acyclic analogue, N-(5-azido-6-chloropyridin-3-ylmethyl)-N'-methyl-N' '-nitroguanidine (2), were prepared in good yields as candidate photoaffinity probes for mammalian and insect nicotinic acetylcholine receptors (nAChRs). The essential intermediate was 5-azido-6-chloropyridin-3-ylmethyl chloride (3) prepared in two ways: from 6-chloro-5-nitronicotinic acid by selective reduction and then diazotization, and from N-(6-chloropyridin-3-ylmethyl)morpholine by an electrophilic azide introduction with lithium diisopropylamide followed by chlorine substitution of morpholine with ethyl chloroformate. Coupling of 3 with 2-nitroiminoimidazolidine gave 1. Conversion of 3 to 2 was achieved in good yields via the hexahydrotriazine intermediate 14. Fortuitously, the azido substituent in 1 and 2 increases the affinity 7-79-fold for rat brain and recombinant alpha4beta2 nAChRs (K(i)s 4.4-60 nM competing with [(3)H](-)-nicotine) while maintaining high potency on both insect nAChRs (Drosophila and Myzus) (K(i)s 1-15 nM competing with [(3)H]imidacloprid). Azidopyridinyl compounds 1 and 2 are therefore candidate photoaffinity probes for characterization of both mammalian and insect receptors.

A locust DNA-binding protein involved in gene regulation by juvenile hormone.

Although juvenile hormone (JH) has essential roles in insect development and reproduction, the molecular mechanisms of gene regulation by JH remain an enigma. In Locusta migratoria, the partially palindromic 15-nt sequence, GAGGTTCGAG(A)/(T)CCT(T)/(C), found upstream of a JH-induced gene, jhp21, was designated as a putative juvenile hormone response element (JHRE). When JH-deprived adult female locusts were treated with the active JH analog, methoprene, a fat body nuclear factor that bound specifically to JHRE appeared after 24 h. Binding exhibited a preference for an inverted repeat with GAGGTTC in the left half-site, a single nucleotide spacer, and a right half-site in which some variation is acceptable. Binding to JHRE was abolished by phosphorylation catalyzed by a C-type protein kinase present in the nuclear extracts. The DNA-binding protein is thus believed to be a transcription factor, which is brought to an active state through the action of JH and then participates in the regulation of certain JH-dependent genes.

HPLC method for the analysis of acetonides of ecdysteroids providing structural information on different vicinal diols

The polyhydroxy structure of ecdysteroids provides a basis for the formation of various derivatives including the acetonides that have been frequently prepared mainly in preparative work. Acetonides can be formed between vicinal diols in cis position such as positions C-2/C-3 and C-20/C-22 in 20-hydroxyecdysone, for example. We have modified an established procedure for the preparation of acetonides, employing a higher concentration of phosphomolybdic acid as a catalyst, and developed a reverse-phase HPLC method to analyze the products on a micro scale. The derivatization method is fast, taking minutes or less, and does not produce artifacts. Thirteen structurally diverse ecdysteroids were studied in detail concerning speed of acetonide formation, and the number and type of intermediate and end products. Mass spectrometry coupled to HPLC was employed to confirm and/or identify ecdysteroids and their acetonide derivatives. Analysis of the chromatographic profiles revealed that detailed information on vicinal diols of ecdysteroids can be derived from the differences in retention times (Deltar.t.)1 of products relative to the parent steroids on reverse-phase HPLC using a linear water/methanol gradient for elution. The specific Deltar.t. values characterize a product as a monoacetonide, a diacetonide, or a triacetonide. Furthermore, this chromatographic shift upon acetonide formation is significantly different among the various monoacetonides and, thus, specific for the site on the ecdysteroid molecule. The same seems to hold for diacetonides. As variation of the Deltar.t. values for a specific type of acetonide was rather small among the tested ecdysteroids, the results should provide reliable structural information. Muristerone A did not completely fit this common pattern. However, this may specify its uncommon ecdysteroid structure. The method has recently been successfully employed to solve the structures of new ecdysteroid metabolites. Arch. Copyright 1999 Wiley-Liss, Inc.

Comparative studies of suidatrestin, a specific inhibitor of trehalases.

Suidatrestin, isolated from a Streptomyces strain, was characterized as a new trehalase inhibitor. Its inhibitory potential was 7 to 50-fold higher than that of validamycin when tested against insect, fungal and mammalian trehalases. The kinetic properties of suidatrestin were studied in vitro with trehalases from flight muscle mitochondria of the fly, Protophormia terraenovae, from larval midgut of the moth, Spodoptera littoralis, and from porcine kidney, as well as with maltase from yeast. Suidatrestin was inactive on maltase but inhibited all trehalases with IC50 values of 0.08-0.1 microM; Ki values ranged from 0.02 to 0.05 microM. The very low Ki/K(m) ratios (3.9 x 10(-6) -4.9 x 10(-6)) indicated excellent in vitro inhibitory action of suidatrestin. When injected into larvae of S. littoralis, suidatrestin required high and repetitive doses which lead to reversible inhibition of larval growth only. Consecutive omission of the inhibitor even stimulated weight increase above that of controls. Significant mortality was achieved at a rather high dose only. Injection of a growth-inhibiting dose of suidatrestin did not change hemolymph osmolality as a measure of sugar concentration. The discrepancy between in vitro and in vivo potency of suidatrestin may be understood once its chemical structure is fully known.

Metabolism of diafenthiuron by microsomal oxidation: procide activation and inactivation as mechanisms contributing to selectivity.

The thiourea insecticide/acaricide diafenthiuron represents a biologically inactive propesticide that requires transformation into the active carbodiimide derivative. The carbodiimide inhibits mitochondrial respiration by selective and covalent binding to the proteolipid (8 kDa) of Fo-ATPase in the inner membrane and to porin (30 kDa) in the outer membrane. The thiourea can be activated by light as well as by cytochrome P450 in the insect. To get insight into the enzymatic mechanisms of activation, model in vitro studies were performed using [14C]diafenthiuron and microsomes from various vertebrate livers and from locust Malpighian tubules. Though there was a common set of metabolites, their quantities varies significantly with the species and assay conditions. As a typical product, p-hydroxydiafenthiuron was identified in assay with rat and mouse microsomes. The sulfomonoxide predominated in hen and fish assays, whereas pig and bovine microsomes almost exclusively produced the carbodiimide. The sulfoxide was shown to be a precursor of the carbodiimide. Formation of all metabolites was dependent on the presence of NADPH and active microsomes. The effects of inhibitors and the requirement for NAPDH suggested a role of cytochrome P450-dependent monooxygenase(s) in the formation of both the hydroxylated product and the carbodiimide. FAD-dependent monooxygenases (FMOs) may also be involved in a step following sulfoxidation. These in vitro studies revealed potential mechanisms contributing to biological selectivity of the effects of a pesticide that acts in a non-selective mode at a conserved mitochondrial site.

Chemistry and biology of thiamethoxam: a second generation neonicotinoid.

Thiamethoxam is the first commercial neonicotinoid insecticide from the thianicotinyl subclass. It was discovered in the course of our optimisation program on neonicotinoids started in 1985. Novel variations of the nitroimino-heterocycle of imidacloprid led to 4-nitroimino-1,3,5-oxadiazinanes exhibiting high insecticidal activity. Among these, thiamethoxam (CGA 293433) was identified as the best compound and selected for worldwide development. The compound can be synthesised in only a few steps and high yield from easily accessible starting materials. Thiamethoxam acts by binding to nicotinic acetylcholine receptors. It exhibits exceptional systemic characteristics and provides excellent control of a broad range of commercially important pests, such as aphids, jassids, whiteflies, thrips, rice hoppers, Colorado potato beetle, flea beetles and wireworms, as well as some lepidopteran species. In addition, a strong preventative effect on some virus transmissions has been demonstrated. Thiamethoxam is developed both for foliar/soil applications and as a seed treatment for use in most agricultural crops all over the world. Low use rates, flexible application methods, excellent efficacy, long-lasting residual activity and favourable safety profile make this new insecticide well-suited for modern integrated pest management programmes in many cropping systems.

[Structure of outpatient pain therapy in Germany. Results of a survey]. / Struktur der ambulanten Schmerztherapie in Deutschland. Ergebnisse einer Umfrage.

Every physician should be able to treat pain regardless of the specialty, but patients with a risk of chronification or chronic pain should receive care from specialized physicians and non-medical professionals. Specialized pain treatment is an additional qualification in Germany, which may be achieved in different specialties by defined structure criteria and experience. The German Society for the Study of Pain and the Professional Association of the German Society of Anesthetists conducted a survey on specialized outpatient pain treatment settings in Germany, encompassing personal and technical equipment, procedures and interdisciplinary multi-professional cooperation. The survey showed a clear increase in the number of pain treatment settings compared to previous surveys, but with a huge span from small single practice or outpatient services at hospitals to large specialized hospitals. However, the quality criteria suggested by the pain treatment societies were not always met. Treatment options for patients with a risk of chronification and chronic pain show regional variations and are insufficiently developed.

The anti-microbial activity of electrolysed oxidizing water against microorganisms relevant in veterinary medicine.

Standards of the German Association of Veterinary Medicine (DVG) for the evaluation of chemical disinfectants were used to assess the anti-microbial efficacy of electrolysed oxidizing water (EOW). Enterococcus faecium, Mycobacterium avium subspecies avium, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans were exposed to anode EOW (pH, 3.0+/-0.1; oxidation-reduction potential (ORP), +1100+/-50 mV; free chlorine, 400+/-20 mg/l Cl2) and combined EOW (7:3 anode:cathode, v/v; pH, 8.3+/-0.1; ORP, 930-950 mV; free chlorine, 271+/-20 mg/l Cl2). In water of standardized hardness (WSH), all bacterial strains were completely inactivated by a 30 min exposure to maximum 10.0% anode EOW (approximately 40.0 mg/l Cl2) or 50.0% combined EOW (approximately 135.5 mg/l Cl2). The sensitivity ranking order for anode EOW to the bacterial test strains was P. mirabilis>S. aureus>M. avium ssp. avium>E. faecium>P. aeruginosa. P. mirabilis and S. aureus decreased to undetectable levels after 5 min of exposure to 7.5% anode EOW (approximately 30.0 mg/l Cl2). Candida albicans was completely inactivated by a 5-min exposure to 5.0% anode EOW. Both, anode and combined EOW exhibited no anti-microbial activities in standardized nutrient broth or after addition of 20.0% bovine serum to the WSH. Further research is necessary to evaluate the efficacy of EOW as a disinfectant under operating conditions in animal production facilities.

Isolation of beta, beta-caroten-2-ol from an insect, Cerura vinula (Lepidoptera).

The monohydroxy carotenoid from the moth Cerura vinula has been shown to be beta, beta-caroten-2-ol on the basis of electronic, infrared, proton magnetic-resonance, and mass spectra. On acid treatment in the presence of molecular oxygen this carotenoid is dehydrogenated to 4',5-retro-beta,beta-caroten-2-one. The identification of beta,beta-caroten-2-ol by its retro product, its time course of acetylation, and its chromatographic properties relative to beta,beta-caroten-3-ol and beta,beta-caroten-4-ol is discussed. This is the first demonstration of a 2-hydroxylated carotenoid in an insect. Implications on the biogenesis of this pigment are considered.

[Identification of beta, beta-caroten-2-ol and beta, beta-carotene-2,2'-diol in the stick insect, Carausius morosus Br.; a reinvestigation study].

Two major carotenoids of the stick insect were reinvestigated and shown to be beta,beta-caroten-2-ol and beta,beta-carotene-2,2'-diol and not isocryptoxanthin (beta,beta-carotin-4-ol) and isozeaxaanthin (beta,beta-carotene-4,4'-diol). Both pigments are esterified with fatty acids. The identification is based on cochromatography with authentic 2-, 3- and 4-isomers of the mono- and dihydroxy pigment, mass spectra, and chemical behaviour. In acid solution beta-beta-carotene-2,2'-diol is specifically dehydrogenated and rearranged to ketones with retro structures in analogy to the reactionof beta,beta-caroten-2-ol as recently reported. The final product of the diol is 4,5-dihydro-4,5'-retro-beta,beta-carotene-2,2'-dione. This is the first demonstration of beta,beta-carotene-2,2'-diol in an animal.