Classification of Cannabis Strains Based on their Chemical Fingerprint-A Broad Analysis of Chemovars in the German Market.

Introduction: Cannabis cultivars were usually categorized based on their genetic profile as sativa, indica, or hybrid types. However, these three criteria do not allow sufficient differentiation between the numerous varieties of cannabis strains. Furthermore, this classification is based on morphological and bio-geographical properties of the plants and does not represent the chemical composition of different cultivars. The concentration of cannabinoids and terpenes are crucial for the pharmacological effect, not only because of the known entourage effect, and therefore needs to be considered by categorization. Materials and Methods: A total of 140 medicinal cannabis flowers available on the German market were analyzed regarding their individual terpene profile using GC-MS analysis. Statistical evaluation was performed to investigate correlations and data relations as well as for clustering. Results: Multivariate analysis showed correlations between individual terpenes. However, there was no statistical correlation between terpene profiles and their respective genetic profile. Terpene profiles of sativa, indica, and hybrid strains are quite heterogenous and clearly showed that there is no relation between terpenes and the estimated pharmacological effect. As a result, we suggest a new classification system based on individual terpene profiles to faster a comprehensive understanding of the expected medical effect. Discussion: Considering main terpenes, we established a concept of six clusters with various terpene profiles being attributed to different medicinal applications. We excluded tetrahydrocannabinol (THC) and cannabidiol (CBD) content from clustering as most of the strains were THC dominant and therefore distort the results. Our pattern of strains with similar terpene profiles might refine the existing classes of chemotypes with different THC:CBD content. Conclusion: The categorization of cannabis strains based on their terpene profiles allows a clearer, finer and, above all, more meaningful classification than the existing sativa/indica classification. Due to the entourage effect and the interactions between cannabinoids and terpenes, this group of substances is also given the necessary consideration when selecting the right medicine for the individual. Within the next steps, further studies are needed with the aim of mapping clinical validated effects to our chemovars. If it is possible to correlate therapy of symptoms to specific chemical profiles personalized cannabinoid therapy will be possible.

Proteolytic cleavage of CD25, the alpha subunit of the human T cell interleukin 2 receptor, by Der p 1, a major mite allergen with cysteine protease activity.

Recent reports have indicated that the cysteine protease activity of Der p 1 may play a significant role in its ability to elicit IgE antibody responses, mainly through cleavage of membrane CD23 on B cells and interleukin (IL)-4 synthesis and secretion from mast cells and basophils. Here we demonstrate for the first time that Der p 1 also cleaves the alpha subunit of the IL-2 receptor (IL-2R or CD25) from the surface of human peripheral blood T cells and, as a result, these cells show markedly diminished proliferation and interferon gamma secretion in response to potent stimulation by anti-CD3 antibody. Given that the IL-2R is pivotal for the propagation of Th1 cells, its cleavage by Der p 1 may consequently bias the immune response towards Th2 cells, thereby creating an allergic microenvironment.

The cysteine protease activity of the major dust mite allergen Der p 1 selectively enhances the immunoglobulin E antibody response.

The house dust mite Dermatophagoides pteronyssinus allergen Der p 1 is the most immunodominant allergen involved in the expression of dust mite-specific immunoglobulin (Ig)E-mediated hypersensitivity. The reason for this potent IgE-eliciting property of Der p 1 remains unknown, but there is mounting in vitro evidence linking the allergenicity of Der p 1 to its cysteine protease activity. Here we demonstrate for the first time that immunization of mice with proteolytically active Der p 1 results in a significant enhancement in total IgE and Der p 1-specific IgE synthesis compared with animals immunized with Der p 1 that was irreversibly blocked with the cysteine protease inhibitor E-64. We conclude that the proteolytic activity of Der p 1 is a major contributor to its allergenicity.

Visfatin/PBEF/Nampt and resistin expressions in circulating blood monocytes are differentially related to obesity and type 2 diabetes in humans.

Low-grade inflammation is important in the development of obesity related pathologies such as insulin resistance and type 2 diabetes, and also cardiovascular disease. Visfatin/PBEF/Nampt and resistin are proinflammatory adipokines secreted from adipocytes, monocytes, and macrophages, and have been linked to atherosclerotic plaque formation, recently. The aim of the present study was to investigate if the expression of these molecules in circulating blood monocytes is altered in obese and/or type 2 diabetic human subjects. Monocytes were isolated by CD14-antibody based magnetic cell sorting from blood samples of 17 lean controls, 20 obese nondiabetic subjects, and 19 obese patients with type 2 diabetes. FACS analysis was performed to test purity of the cell preparations. Expression of the different adipokines was measured by multiplex real-time PCR on RNA-level. Visfatin/PBEF/Nampt was found to be very strongly expressed in monocytes, whereas resistin levels were significantly lower. Furthermore, visfatin/PBEF/Nampt expression was significantly upregulated in obese type 2 diabetic patients, whereas obese nondiabetics exhibited similar levels compared to lean controls, indicating that visfatin/PBEF/Nampt levels are related to type 2 diabetes rather than to obesity. In contrast, resistin expression displayed a different pattern being significantly increased in obese subjects compared to controls but not related to type 2 diabetes. These data suggest a differential role for these two proinflammatory adipokines in linking metabolic diseases to atherosclerosis with visfatin/PBEF/Nampt being more important in patients with type 2 diabetes and resistin in obese but nondiabetic human subjects.

Role of TLR3 in the immunogenicity of replicon plasmid-based vaccines.

Replicon plasmids encoding an alphavirus RNA replicase constitute an alternative to conventional DNA plasmids with promise for DNA vaccination in humans. Replicase activity amplifies the levels of transgene mRNA through a copying process involving double-stranded (ds) RNA intermediates, which contribute to vaccine immunogenicity by activating innate antiviral responses. Toll-like receptor 3 (TLR3) is a dsRNA innate immune receptor expressed by antigen-presenting dendritic cells (DCs). Here, we test the hypothesis that TLR3 is necessary for the immunogenicity of replicon plasmid-based DNA vaccines. We show that mouse CD8 alpha(+) DC phagocytose dying replicon plasmid-transfected cells in vitro and are activated in a TLR3-dependent manner by dsRNA present within those cells. However, we find that cytotoxic T-cell responses to a replicon plasmid intramuscular vaccine are not diminished in the absence of TLR3 in vivo. Our results underscore the potential role of TLR3 in mediating immune activation by dsRNA-bearing replicon plasmid-transfected cells and indicate that other innate sensing pathways can compensate for TLR3 absence in vivo.

Non-classical light emission from a single electrically driven quantum dot.

Easy to handle light sources with non-classical emission features are strongly demanded in the growing field of quantum communication. We report on single-photon emission from an electrically pumped quantum dot with unmatched spectral purity, making spatial or spectral filtering dispensable.

Urgent request on avian influenza.

Highly pathogenic avian influenza (HPAI) H5N8 is currently causing an epizootic in Europe, infecting many poultry holdings as well as captive and wild bird species in more than 10 countries. Given the clear clinical manifestation, passive surveillance is considered the most effective means of detecting infected wild and domestic birds. Testing samples from new species and non-previously reported areas is key to determine the geographic spread of HPAIV H5N8 2016 in wild birds. Testing limited numbers of dead wild birds in previously reported areas is useful when it is relevant to know whether the virus is still present in the area or not, e.g. before restrictive measures in poultry are to be lifted. To prevent introduction of HPAIV from wild birds into poultry, strict biosecurity implemented and maintained by the poultry farmers is the most important measure. Providing holding-specific biosecurity guidance is strongly recommended as it is expected to have a high impact on the achieved biosecurity level of the holding. This is preferably done during peace time to increase preparedness for future outbreaks. The location and size of control and in particular monitoring areas for poultry associated with positive wild bird findings are best based on knowledge of the wider habitat and flight distance of the affected wild bird species. It is recommended to increase awareness among poultry farmers in these established areas in order to enhance passive surveillance and to implement enhanced biosecurity measures including poultry confinement. There is no scientific evidence suggesting a different effectiveness of the protection measures on the introduction into poultry holdings and subsequent spread of HPAIV when applied to H5N8, H5N1 or other notifiable HPAI viruses.

Two strategies to prepare neural cortical cytoskeleton components for the generation of monoclonal antibodies.

Like most other cells, neurons possess a spectrin/actin based network closely associated with the inner side of the cell membrane, the cortical cytoskeleton. This structure serves many diverse functions during axonal outgrowth. In the growth cone, the cortical cytoskeleton is involved in surface shaping, modulation of integral membrane proteins, and signal transduction. We developed two strategies to prepare material enriched for neural cortical cytoskeleton. The first strategy combined the isolation of a membrane/cortical cytoskeleton fraction by density gradient centrifugation with an enzymatic degradation of cell surface proteins. The second strategy is based on the attachment and crosslinking of single cells to beads, allowing for the removal of the cell contents by cell disruption; only membrane/cortical cytoskeleton patches are retained on the beads. Both strategies made use of the intimate association of the cortical cytoskeleton with the cell membrane, permitting the removal of cytoplasm, organelles and cytoplasmic cytoskeleton while retaining the cortical cytoskeleton. Monoclonal antibodies generated using both preparations as immunization material were screened for recognition of intracellular structures in axons and growth cones of retinal ganglion cells in culture. A quantitative specification of the antibodies is presented and six antibodies are characterized in immunolabelings and Western blot analysis.

Gene expression of receptors and enzymes involved in GABAergic and glutamatergic neurotransmission in the CNS of rats behaviourally dependent on ethanol.

The steady state levels of the messenger RNA (mRNA) of eight GABA(A) receptor subunits, five glutamate receptor subunits and seven enzymes involved in the synthesis of glutamate and GABA were measured in eight regions of rat brain in a recently developed animal model of 'behavioural dependence' on ethanol. 'Behavioural dependence' including loss of control was induced by offering the rats the choice between ethanol and water over a 9-month period (Group A). This group was compared with a group given the choice between ethanol and water for only 2 months (not yet 'behaviourally dependent', Group B), a group forced to consume ethanol as sole fluid over a 9-month period (also not 'behaviourally dependent', Group C) and ethanol-naive control rats (Group D). All groups were sacrificed 1 month after the ethanol was withdrawn. The mRNA concentrations of all eight GABA receptor subunits, four out of the five subunits of different glutamate receptors and those of seven enzymes involved in GABA and glutamate production were reduced almost exclusively in the parieto-occipital cortex in Groups A and B, but not Group C. These data suggest that the synthesis of glutamate and GABA and the activities of their respective neurons are selectively impaired in the parieto-occipital cortex in the groups having consumed ethanol in a free-choice design, in which its rewarding properties can better take effect than after forced administration. As the parieto-occipital cortex is believed to contain emotional memory structures, it may be hypothesized that the glutamatergic and GABAergic neuronal systems in this area are involved in the development of memory for reward from ethanol. However, they are not specifically associated with 'behavioural dependence'.

Dopamine receptor gene expression in an animal model of 'behavioral dependence' on ethanol.

The steady-state levels of messenger RNA (mRNA) of five cloned dopamine (D) receptors were measured in five brain regions in rats in a recently developed animal model of 'behavioral dependence' on ethanol. One group of rats was given the choice between ethanol and water over a 9-month period and developed 'behavioral dependence' on ethanol (group a). This group was compared with a group given the choice between ethanol and water for only 2 months (not yet behaviorally dependent, group b), a group forced to consume ethanol as sole fluid over a 9-month period (not behaviorally dependent, group c) and ethanol-naive control rats. All groups were sacrificed 1 month after ethanol withdrawal. The concentrations of mRNA of D3-receptors in the limbic forebrain (which included the nucleus accumbens) were significantly lowered in groups a and b, but unchanged in group c. D3 mRNA levels were reduced in the hippocampus of group b and unchanged in the cortex, amygdala and striatum. No significant changes in the mRNA concentrations of D1-, D2-, D4- or D5-receptors were seen in the five brain regions in any group. In conclusion, chronic consumption of ethanol under the 'free-choice condition', which may best induce the drug-rewarding effect, leads to specific changes in the D3-receptor gene expression which were not seen after forced ethanol administration. Changes in D3 mRNA levels were, however, not a specific correlate of 'behavioral dependence', as they were also detected in rats not yet 'behaviorally dependent' (group b).

Riboflavin can decrease the nephrotoxic effect of chromate in young and adult rats.

The influence of 5 mg vitamin B2/100 g b.wt. (B2, riboflavin) on the nephrotoxic effect of 1 or 2 mg Na2Cr2O7/100 g b.wt. (Cr) was investigated in 55- and 10-day-old rats, respectively. Nephrotoxic effect was evaluated by the determination of urinary volume and protein excretion as well as the concentration of blood urea nitrogen (BUN). The concomitant administration of Cr and B2 only in 55-day-old rats increased the nephrotoxicity shown by enhanced proteinuria and BUN. B2, administered 3 h after Cr, was able to diminish Cr nephrotoxicity significantly in 55- and 10-day-old rats. The effect of B2 on Cr nephrotoxicity could be interpreted not by the stimulatory effect of B2 on GSSG reductase, which was abolished by Cr; but by its antioxidant effect.

Influence of muscle forces on femoral strain distribution.

Musculoskeletal loading influences the stresses and strains within the human femur and thereby affects the processes of bone modeling and remodeling. It is essential for implant design and simulations of bone modeling processes to identify locally high or low strain values which may lead to bone resorption and thereby affect the clinical outcome. Using a finite element model the stresses and strains of a femur with all thigh muscle and joint contact forces were calculated for four phases of a gait cycle. Reduced load sets with only a few major muscles were analyzed alternatively. In a completely balanced femur with all thigh muscles the stress and strain patterns are characterized by combined bending and torsion throughout the bone. Similar to in vivo recordings, the model with all thigh muscles showed peak surface strains below 2000 mu epsilon (45% gait cycle). Under simplified load regimes surface strains reached values close to 3000 mu epsilon. Within the proximal femur, the simplified load regimes produced differences in strain as high as 26% in comparison to those with all thigh muscles included. This difference is reduced to 5% if the adductors are added to a loading consisting of hip contact, abductors and ilio-tibial band. This study demonstrates the importance of an ensemble of muscle forces to reproduce a physiological strain distribution in the femur. Analytical attempts to simulate bone modeling, remodeling or bone density distributions should therefore rely on fully balanced external load regimes which account for the role of the various soft tissue forces.

[Interrelationships between methylmercury exposure and selenium supplementation in cockerels using selected laboratory diagnostic parameters and toxicological parameters of residues]. / Wechselwirkungen zwischen Methylquecksilber-Belastung und Selen-Supplementation bei Junghähnen anhand ausgewählter labordiagnostischer und rückstandstoxikologischer Parameter.

A feed loading experiment was applied in 2 phases to 45 young cocks over 12 weeks, using 1,2-N,N-bis(methylmercury)-p-toluolsulphamide-dressed wheat (50% of base ration). Investigations were conducted to study the effects of selenium supplementation (0,2 mg Se as sodium selenite/l drinking water) on biochemical and hematological parameters (calcium, phosphorus, total protein, albumin, creatinine, urea, activity of alkaline phosphatase, hematocrit, hemoglobin, leucocyte count) as well as on parameters relating to toxicological residues (selenium and mercury levels in liver, musculature and kidneys). Statistically secured differences were found to exist between the experimental groups with regard to selenium and mercury in the liver and mercury concentrations in kidneys. Possible interrelationships were discussed.

Hypertrophy of infected Peyer's patches arises from global, interferon-receptor, and CD69-independent shutdown of lymphocyte egress.

Lymphoid organ hypertrophy is a hallmark of localized infection. During the inflammatory response, massive changes in lymphocyte recirculation and turnover boost lymphoid organ cellularity. Intriguingly, the exact nature of these changes remains undefined to date. Here, we report that hypertrophy of Salmonella-infected Peyer's patches (PPs) ensues from a global "shutdown" of lymphocyte egress, which traps recirculating lymphocytes in PPs. Surprisingly, infection-induced lymphocyte sequestration did not require previously proposed mediators of lymphoid organ shutdown including type I interferon receptor and CD69. In contrast, following T-cell receptor-mediated priming, CD69 was essential to selectively block CD4(+) effector T-cell egress. Our findings segregate two distinct lymphocyte sequestration mechanisms, which differentially rely on intrinsic modulation of lymphocyte egress capacity and inflammation-induced changes in the lymphoid organ environment.

Dipeptidyl-peptidase 4 and attractin expression is increased in circulating blood monocytes of obese human subjects.

Dipeptidyl-peptidase (DPP)-4, which catalizes the degradation of the insulinotropic incretin glucagon-like-peptide (GLP)-1, and the DPP-4 like enzyme attractin are involved in activation of T-lymphocytes and monocytes. Recently, it has been demonstrated, that the risk for certain infections is increased in type 2 diabetic patients under DPP-4 inhibitor treatment. The aim of the present study was to examine the expression of DPP-4 and attractin in circulating blood monocytes of obese and type 2 diabetic subjects. Monocytes were isolated by CD14-antibody based magnetic cell sorting from blood samples of 17 lean controls, 20 obese, non-diabetic subjects and 19 obese patients with type 2 diabetes. FACS analysis was performed to test purity of the cell preparations. Expression was measured by multiplex RT-PCR on RNA-level. DPP-4 and attractin were detectable in human circulating monocytes with attractin being expressed at higher levels compared to DPP-4. Both enzymes were significantly higher expressed in circulating blood monocytes of obese subjects compared to lean controls. In contrast, type 2 diabetes did not significantly affect expression levels. Finally, neither DPP-4 nor attractin expression was altered by sitagliptin or insulin treatment. In conclusion, our data demonstrate, that expressions of DPP-4 and attractin in circulating blood monocytes of human subjects are influenced by metabolic abnormalities with obesity being an important factor.

A probabilistic effect assessment model for hazardous substances at the workplace.

A major problem in risk assessment is the quantification of uncertainties. A probabilistic model was developed to consider uncertainties in the effect assessment of hazardous substances at the workplace. Distributions for extrapolation factors (time extrapolation, inter- and intraspecies extrapolation) were determined on the basis of appropriate empirical data. Together with the distribution for the benchmark dose obtained from substance-specific dose-response modelling for the exemplary substances 2,4,4-trimethylpentene (TMP) and aniline, they represent the input distributions for probabilistic modelling. These distributions were combined by Monte Carlo simulation. The resulting target distribution describes the probability that an aspired protection level for workers is achieved at a certain dose and the uncertainty associated with the assessment. In the case of aniline, substance-specific data on differences in susceptibility (between species; among humans due to genetic polymorphisms of N-acetyltransferase) were integrated in the model. Medians of the obtained target distributions of the basic models for TMP and aniline, but not of the specific aniline model are similar to deterministically derived reference values. Differences of more than one order of magnitude between the medians and the 5th percentile of the target distributions indicate substantial uncertainty associated with the effect assessment of these substances. The probabilistic effect assessment model proves to be a practical tool to integrate quantitative information on uncertainty and variability in hazard characterisation.

Application of prominent spectral lines in the 125-180 nm range for inductively coupled plasma optical emission spectrometry.

A new axially viewed ICP optical emission spectrometer featuring an argon-filled optic and CCD detectors was evaluated for the application of prominent spectral lines in the 125-180 nm range. This wavelength range was investigated for several analytical applications of inductively coupled plasma optical emission spectrometry (ICP-OES). There are different advantages for the application of spectral lines below 180 nm. A number of elements, such as Al, Br, Cl, Ga, Ge, I, In, N, P, Pb, Pt, S and Te, were found to have the most intense spectral lines in the wavelength range from 125-180 nm. Compared with lines above 180 nm higher signal-to-background ratios were found. Low limits of detection using pneumatic nebulization of aqueous solutions for sample introduction were calculated for Al II 167.080 nm (0.04 microg L(-1)), Br I 154.065 nm (9 microg L(-1)), Cl I 134.724 nm (19 microg L(-1)), Ga II 141.444 nm (0.8 microg L(-1)), Ge II 164.919 nm (1.3 microg L(-1)), II 142.549 nm (13 microg L(-1)), In II 158.583 nm (0.2 microg L(-1)), P I 177.500 nm (0.9 microg L(-1)), Pb II 168.215 nm (1.5 microg L(-1)), Pt II 177.709 nm (2.6 microg L(-1)), S I 180.731 nm (1.9 microg L(-1)) and Te I 170.00 nm (4.6 microg L(-1)). Numerous application examples for the use of those lines and other important spectral lines below 180 nm are given. Because of fewer emission lines from transition elements, such as Fe, Co, Cr, lines below 180 nm often offer freedom from spectral interferences. Additional lines of lower intensity for the determination of higher elemental concentrations are also available in the vacuum ultraviolet spectral range. This is specially useful when the concentrations are not in the linear range of calibration curves obtained with commonly used lines.

[Metabolism and hematology studies of intensively raised fattened lambs under various feeding and management conditions. 2. The effect of various management methods on the blood picture, serum mineral and bilirubin concentration and vitamin A concentration in the liver]. / Stoffwechsel- und hämatologische Untersuchungen an industriemässig gehaltenen Mastlämmern bei unterschiedlichen Fütterungs- und Haltungsbedingungen. 2. Der Einfluss unterschiedlicher Haltungsvarianten auf das Blutbild, den Mineralstoff- und Bilirubingehalt im Serum und den Vitamin-A-Gehalt der Leber

Calcium, phosphorus, magnesium and bilirubin content, blood picture and liver vitamin A were studied in four groups of intensively fattened lambs kept under different conditions of management, in comparison with a control group. Results are shown in graphs and tables, Group-specific differences were found in some of the parameters examined: - Ca, P, erythrocytes, leucocytes, vitamin A.

[Metabolism and hematology studies of intensively raised fattened lambs under various feeding and management conditions. 1. The effect of various feeding methods on the blood picture, serum mineral and bilirubin concentration, and vitamin A concentration in the liver]. / Stoffwechsel- und hämatologische Untersuchungen an industriemässig gehaltenen Mastlämmern bei unterschiedlichen Fütterungs- und Haltungsbedingungen. 1. Der Einfluss unterschiedlicher Fütterungsvarianten auf das Blutbild, den Mineralstoff- und Bilirubingehalt im Serum und den Vitamin-A-Gehalt der Leber

Calcium, phosphorus, magnesium and bilirubin content, blood picture and liver vitamin A were determined in six groups of lambs fattened on different rations, in comparison with a control group. The results are presented in graphs and tables. There was no clear difference between the various groups, in the criteria examined.