Pharmacological effects of acute and repeated administration of Delta(9)-tetrahydrocannabinol in adolescent and adult rats

Adolescents of many mammalian species exhibit rapid physiological change that is accompanied by behaviors such as increased risk taking and social interaction with peers. Marijuana abusers frequently report that their initial use occurred during adolescence. Our goal was to determine whether the in vivo effects of Delta(9)-tetrahydrocannabinol (Delta(9)-THC) differed in adolescent and adult rats. Following initial testing with Delta(9)-THC in adolescent [postnatal day (PN)29] and adult (>PN60) rats of both sexes, we injected rats twice daily with 10 mg/kg Delta(9)-THC or vehicle for 9.5 days. Subsequently, rats were again injected with their initial dose of Delta(9)-THC and tested. In all rats, Delta(9)-THC produced dose-dependent locomotor suppression, antinociception, hypothermia and catalepsy. Some age-dependent differences in potency and efficacy were noted. Although Delta(9)-THC dose-effect functions were more similar across age after repeated exposure, subchronic dosing produced greater change in the hypothermic and locomotor effects of Delta(9)-THC in adolescents, but less change in its antinociceptive effects. These results suggest that the effects of initial exposure to Delta(9)-THC may not be entirely predictive of the effects of repeated exposure. Despite similarities in pharmacological effects of Delta(9)-THC after repeated use, adolescents and adults may exhibit differences in the pattern of transition from use to abuse.     

Problems with current recommendations for susceptibility testing of Haemophilus influenzae

We compared results of MIC and disk susceptibility tests on Haemophilus test medium (HTM) and those on comparative media. Ampicillin MICs were determined with seven ampicillin-resistant, non-beta-lactamase-producing (AmprNBLP) isolates by using HTM and supplemented brain heart infusion (sBHI) agar. Ampicillin and amoxicillin-clavulanate disk tests with 16 AmprNBLP strains, 18 ampicillin-susceptible (Amps) isolates, and 17 ampicillin-resistant, beta-lactamase-producing (AmprBLP) strains were performed by using five media: laboratory-prepared HTM (PHTM), commercial HTM (CHTM), sBHI, enriched chocolate agar, and Mueller-Hinton chocolate agar. We observed that five of seven and three of seven AmprNBLP strains were misclassified as susceptible with PHTM (MIC, less than 2 micrograms/ml) with inocula of 10(3) and 10(5) CFU, respectively, but were resistant with sBHI (MIC, greater than or equal to 2 micrograms/ml). Whereas Mueller-Hinton chocolate agar and enriched chocolate agar plates supported the growth of all 51 strains by the disk tests, 37% (19 of 51) and 8% (4 of 51) of strains did not grow on PHTM and CHTM, respectively. Lack of growth on PHTM was observed for all three phenotypes; 7 of 18 Amps, 4 of 17 AmprBLP, and 8 of 16 AmprNBLP strains did not grow. The four strains that did not grow on CHTM were all AmprNBLP isolates. Zone sizes were significantly larger on PHTM than on the other media. Of the strains that were evaluable by the new National Committee for Clinical Laboratory Standards guidelines with either PHTM or CHTM, all Amps strains were classified as susceptible. Among the AmprBLP strains, CHTM correctly identified all as resistant, whereas PHTM detected two isolates to be intermediate. Among the AmprNBLP strains, CHTM and PHTM misclassified four (33%) and five (62%) isolates, respectively, as susceptible; an additional isolate was identified as intermediate on both media. We conclude that there is strain-dependent growth on HTM, that adoption of this medium for routine Haemophilus susceptibility testing is problematic due to this growth variability, and that detection of AmprNBLP isolates would be unreliable.     

In vivo characterization of the highly selective monoacylglycerol lipase inhibitor KML29: antinociceptive activity without cannabimimetic side effects

Background and PurposeSince monoacylglycerol lipase (MAGL) has been firmly established as the predominant catabolic enzyme of the endocannabinoid 2-arachidonoylglycerol (2-AG), a great need has emerged for the development of highly selective MAGL inhibitors. Here, we tested the in vivo effects of one such compound, KML29 (1,1,1,3,3,3-hexafluoropropan-2-yl 4-(bis(benzo[d][1,3]dioxol-5-yl)(hydroxy)methyl)piperidine-1-carboxylate).Experimental ApproachIn the present study, we tested KML29 in murine inflammatory (i.e. carrageenan) and sciatic nerve injury pain models, as well as the diclofenac-induced gastric haemorrhage model. KML29 was also evaluated for cannabimimetic effects, including measurements of locomotor activity, body temperature, catalepsy, and cannabinoid interoceptive effects in the drug discrimination paradigm.Key ResultsKML29 attenuated carrageenan-induced paw oedema and completely reversed carrageenan-induced mechanical allodynia. These effects underwent tolerance after repeated administration of high-dose KML29, which were accompanied by cannabinoid receptor 1 (CB₁) receptor desensitization. Acute or repeated KML29 administration increased 2-AG levels and concomitantly reduced arachidonic acid levels, but without elevating anandamide (AEA) levels in the whole brain. Furthermore, KML29 partially reversed allodynia in the sciatic nerve injury model and completely prevented diclofenac-induced gastric haemorrhages. CB₁ and CB₂ receptors played differential roles in these pharmacological effects of KML29. In contrast, KML29 did not elicit cannabimimetic effects, including catalepsy, hypothermia and hypomotility. Although KML29 did not substitute for Δ⁹-tetrahydrocannabinol (THC) in C57BL/6J mice, it fully and dose-dependantly substituted for AEA in fatty acid amide hydrolase (FAAH) (−/−) mice, consistent with previous work showing that dual FAAH and MAGL inhibition produces THC-like subjective effects.Conclusions and ImplicationsThese results indicate that KML29, a highly selective MAGL inhibitor, reduces inflammatory and neuropathic nociceptive behaviour without occurrence of cannabimimetic side effects.Linked ArticlesThis article is part of a themed section on Cannabinoids 2013. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue-6     

The Effect of Green Tea Extract and EGCG on the Signaling Network in Squamous Cell Carcinoma

Green tea and its major active component, epigallocatechin-3-gallate (EGCG), have been reported to have anticancer activity on various cancers. However, the exact molecular mechanism of its anticancer activity is still not well understood. We investigated the anticancer activity of green tea extract (GTE) and EGCG on 3 human squamous carcinoma cell lines (CAL-27, SCC-25, and KB) in vitro. We also examined the effects of GTE and EGCG on cell signaling networks using our newly developed Pathway Array technology, which is an innovative proteomic assay to globally screen changes in protein expression and phosphorylation. Our results demonstrated that GTE and EGCG inhibited all 3 squamous carcinoma cells’ growth via S and G₂/M phase arrest, but different sensitivities to GTE and EGCG in different cell lines were observed: CAL-27 cells were more sensitive to the both agents than SCC-25 and KB cells, and GTE at an EGCG equivalent concentration displayed a stronger inhibition than EGCG alone. The Pathway Array assessment of 107 proteins indicated that different signaling pathways were activated in different cell lines, suggesting heterogeneity at the signaling network level. After treatment with GTE or EGCG, a total of 21 proteins and phosphorylations altered significantly in all 3 cell lines based on analysis of variance (ANOVA) (P < 0.05). The major signaling pathways affected by GTE and EGCG were EGFR and Notch pathways, which, in turn, affected cell cycle-related networks. These results suggested that GTE and EGCG target multiple pathways or global networks in cancer cells, which resulted in collective inhibition of cancer cell growth. The finding pointed out the future direction to study the underlying mechanism of the chemotherapeutic and chemopreventive activities of EGCG and GTE.     

Sources of pertussis infection in young infants: A review of key evidence informing targeting of the cocoon strategy

Background

The relative contribution of different categories of contact in transmitting pertussis to very young infants, who experience the most severe morbidity, is the most important single factor determining the likely benefit of pertussis vaccination of their close contacts (the “cocooning” strategy).

Objective

To identify, evaluate the quality of and summarise existing data on potential sources of infant pertussis infection in high income countries, focussing on infants under 6 months old.Data sources: Online databases MEDLINE and EMBASE. Additional studies were identified from the reference lists of relevant articles.Study selection and analysis: Study quality was evaluated by standardised criteria, based on the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement. Pooled estimates of the proportion of pertussis cases attributable to various contact sources were calculated using data from the highest quality studies.

Results

Nine studies met the inclusion criteria; seven included data on contacts of hospitalised infants less than 6 months old. Case definitions and methods of contact ascertainment were variable. Most identified sources were from the household, of which 39% (95%CI 33–45%) were mothers, 16% (95%CI 12–21%) fathers, and 5% (95%CI 2–10%) grandparents. Estimates for siblings (16–43%) and non-household contacts (4–22%) were more heterogeneous. For 32–52% of infant cases, no source was identified. Asymptomatic pertussis infection was found in 8–13% of contacts evaluated.

Conclusions

These data suggest that the greatest potential impact of pertussis vaccination of adults to prevent severe disease in young infants comes from vaccinating mothers, followed by fathers, with grandparents having a minor role. Siblings varied in importance and, given recent data regarding waning immunity in vaccinated children, need further study. Non-household sources are also well documented, highlighting the potential limitations of the cocoon strategy to prevent severe infant disease.     

Human epidermal and monocyte-derived langerhans cells express functional P2X receptors

Monocyte-derived dendritic cells (Mo-DC) express functional P2X7 receptors; however, the expression of these receptors on tissue-derived dendritic cells including epidermal Langerhans cells (LC) is unknown. Using immunolabeling and flow cytometry, we demonstrated that P2X7 was present on both human epidermal LC and monocyte-derived LC (Mo-LC), as well as on human keratinocytes. The ecto-ATPDase (CD39) was also present on LC, but not keratinocytes. The P2X7 agonists, 2'- and 3'-0(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP) or ATP, but neither adenosine 5'-diphosphate (ADP) nor uridine 5'-triphosphate (UTP), induced ethidium+ uptake into these cells. Furthermore, ATP-induced ethidium+ uptake into epidermal LC, Mo-LC and keratinocytes was inhibited by the specific P2X7 antagonist, KN-62 (1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine). ATP-induced ethidium+ uptake into Mo-LC and Mo-DC was 2- and 3-fold greater, respectively, than that for fresh monocytes. P2X7 activation on LC induced downstream signaling events, as BzATP or ATP, but neither ADP nor UTP, induced shedding of the low-affinity receptor for IgE (CD23) from Mo-LC. This process was inhibited by KN-62. Finally, ATP-induced ethidium+ uptake and CD23 shedding were impaired in Mo-LC obtained from subjects homozygous for the loss-of-function Glu-496 to Ala polymorphism in the P2X7 receptor. These results demonstrate that human LC express functional P2X7 receptors, and suggest a role for this receptor in the skin immune system.     

Advancing the science of mHealth

Mobile health (mHealth) technologies have the potential to greatly impact health research, health care, and health outcomes, but the exponential growth of the technology has outpaced the science. This article outlines two initiatives designed to enhance the science of mHealth. The mHealth Evidence Workshop used an expert panel to identify optimal methodological approaches for mHealth research. The NIH mHealth Training Institutes address the silos among the many academic and technology areas in mHealth research and is an effort to build the interdisciplinary research capacity of the field. Both address the growing need for high quality mobile health research both in the United States and internationally. mHealth requires a solid, interdisciplinary scientific approach that pairs the rapid change associated with technological progress with a rigorous evaluation approach. The mHealth Evidence Workshop and the NIH mHealth Training Institutes were both designed to address and further develop this scientific approach to mHealth.