The 2-alkyl-2<Emphasis Type="Italic">H</Emphasis>-indazole regioisomers of synthetic cannabinoids AB-CHMINACA,AB-FUBINACA,AB-PINACA,and 5F-AB-PINACA are possible manufacturing impurities with cannabimimetic activities

Indazole-derived synthetic cannabinoids (SCs) featuring an alkyl substituent at the 1-position and l-valinamide at the 3-carboxamide position (e.g., AB-CHMINACA) have been identified by forensic chemists around the world, and are associated with serious adverse health effects. Regioisomerism is possible for indazole SCs, with the 2-alkyl-2H-indazole regioisomer of AB-CHMINACA recently identified in SC products in Japan. It is unknown whether this regiosiomer represents a manufacturing impurity arising as a synthetic byproduct, or was intentionally synthesized as a cannabimimetic agent. This study reports the synthesis, analytical characterization, and pharmacological evaluation of commonly encountered indazole SCs AB-CHMINACA, AB-FUBINACA, AB-PINACA, 5F-AB-PINACA and their corresponding 2-alkyl-2H-indazole regioisomers. Both regioisomers of each SC were prepared from a common precursor, and the physical properties, ¹H and ¹³C nuclear magnetic resonance spectroscopy, gas chromatography–mass spectrometry, and ultraviolet–visible spectroscopy of all SC compounds are described. Additionally, AB-CHMINACA, AB-FUBINACA, AB-PINACA, and 5F-AB-PINACA were found to act as high potency agonists at CB₁ (EC₅₀ = 2.1–11.6 nM) and CB₂ (EC₅₀ = 5.6–21.1 nM) receptors in fluorometric assays, while the corresponding 2-alkyl-2H-indazole regioisomers demonstrated low potency (micromolar) agonist activities at both receptors. Taken together, these data suggest that 2-alkyl-2H-indazole regioisomers of AB-CHMINACA, AB-FUBINACA, AB-PINACA, and 5F-AB-PINACA are likely to be encountered by forensic chemists and toxicologists as the result of improper purification during the clandestine synthesis of 1-alkyl-1H-indazole regioisomers, and can be distinguished by differences in gas chromatography–mass spectrometry fragmentation pattern.     

5F-Cumyl-PINACA in ‘e-liquids’ for electronic cigarettes: comprehensive characterization of a new type of synthetic cannabinoid in a trendy product including investigations on the in vitro and in vivo phase I metabolism of 5F-Cumyl-PINACA and its non-fluorinated analog Cumyl-PINACA

Purpose

In recent years e-liquids used in electronic cigarettes have become an attractive alternative to smoking tobacco. A new trend is the use of e-liquids containing synthetic cannabinoids (SCs) instead of smoking cannabis or herbal mixtures laced with SCs. In the frame of a systematic monitoring of the online market of ‘legal high’ products, e-liquids from online retailers who also sell herbal blends were bought.

Methods

The products were analyzed by gas chromatography-mass spectrometry. In some of the e-liquids an unknown compound was detected which was identified as the SC 5F-Cumyl-PINACA (1-(5-fluoropentyl)-N-(2-phenylpropan-2-yl)-1H-indazole-3-carboxamide) by nuclear magnetic resonance analysis. To investigate the phase I metabolism of this new class of compounds, 5F-Cumyl-PINACA and its non-fluorinated analog Cumyl-PINACA were incubated with pooled human liver microsomes (pHLM). Cumyl-PINACA was additionally ingested orally (0.6 mg) by a volunteer in a controlled self-experiment. To assess the relative potency of Cumyl-PINACA a set of SCs were characterized using a cAMP assay.

Results

Metabolism of 5F-Cumyl-PINACA and Cumyl-PINACA showed similarities with AM-2201 and JWH-018. The main metabolites were formed by hydroxylation at the N-pentyl side chain. The main metabolites detected in the volunteer’s urine sample were the same as in the pHLM assay. All SCs tested with the cAMP assay were full agonists at the CB₁ receptor. Cumyl-PINACA was the most potent SC among the tested compounds and showed an EC₅₀ value of 0.06 nM.

Conclusions

The increasing popularity of e-liquids particularly among young people, and the extreme potency of the added SCs, pose a serious threat to public health. To our knowledge, this is the first report describing the tentative identification of human in vivo metabolites of Cumyl-PINACA and 5F-Cumyl-PINACA.

     

∆9-Tetrahydrocannabinol,a major marijuana component,enhances the anesthetic effect of pentobarbital through the CB1 receptor

Purpose

?⁹-Tetrahydrocannabinol (?⁹-THC) and cannabidiol (CBD), major psychoactive constituents of marijuana, induce potentiation of pentobarbital-induced sleep in mice. We have elucidated the mechanism of enhancement of the anesthetic effect of pentobarbital by cannabinoids.

Methods

We carried out pharmacological experiment and cannabinoid₁ (CB₁) receptor binding assay using CB₁ antagonists to clarify whether the CB₁ receptor is involved in the synergism or not. The affinities of cannabinoids for the CB₁ receptor in the mouse brain synaptic membrane were evaluated using a specific CB₁ ligand, [³H]CP55940.

Results

Although the potentiating effect of ?⁹-THC on pentobarbital-induced sleep was attenuated by co-administration of CB₁ receptor antagonists, such as SR141716A and AM251, at a dose of 2 mg/kg, intravenously (i.v.) to mice, the CBD-enhanced pentobarbital-induced sleep was not inhibited by SR141716A. The inhibitory constant (Ki) values of ?⁹-THC and CBD were 6.62 and 2010 nM, respectively, showing a high affinity of ?⁹-THC and a low affinity of CBD for the CB₁ receptor, respectively. A high concentration of pentobarbital (1 mM) did not affect specific [³H]CP55940 binding on the mouse brain synaptic membrane.

Conclusions

These results suggest that binding of ?⁹-THC to the CB₁ receptor is involved in the synergism with pentobarbital, and that potentiating effect of CBD with pentobarbital may differ from that of ?⁹-THC. We successfully demonstrated that ?⁹-THC enhanced the anesthetic effect of pentobarbital through the CB₁ receptor.

     

Evaluation of carboxamide-type synthetic cannabinoids on the functional activities at cannabinoid receptors and biological effects via inhalation exposure test

Purpose

Three synthetic carboxamide-type cannabinoids (5F-MDMB-PICA, 5F-EMB-PINACA, and AMB-CHMICA) were evaluated in terms of their in vitro activities at the cannabinoid receptors CB₁ and CB₂ and in vivo biological effects when smoking the synthetic cannabinoids to assess their biological effects.

Methods

[³⁵S]Guanosine-5′-O-(3-thio)-triphosphate binding assays were performed to investigate the half maximal effective concentration values of the test compounds at the CB₁ and CB₂ receptors. Additionally, the biological effects were evaluated by observing and scoring the behavior of mice with equipment in which they inhaled smoke from a herbal mixture containing the test compounds.

Results

All three synthetic cannabinoids tested in this study activated the CB₁ and CB₂ receptors in vitro. 5F-MDMB-PICA showed less than 1 nM of the half maximal effective concentration value for both receptors. Therefore, it was suggested that 5F-MDMB-PICA was the strongest CB₁ and CB₂ receptor agonist in comparison with synthetic cannabinoids evaluated in the past. The degree of the various biological effects, specifically passivity, spontaneous activity, abnormal gait, abnormal position, and grip strength, when smoking the synthetic cannabinoids corresponded to the functional activity at the CB₁ receptor. However, some biological effects differed between 5F-MDMB-PICA and 5F-MDMB-PINACA, used as a positive control, and AMB-CHMICA induced some biological effects in contrast to the other tested synthetic cannabinoids.

Conclusion

This study provides information regarding the biological effects when smoking synthetic cannabinoids from the functional activities at the CB₁ and CB₂ receptors, considering their way of inhalation and thermal degradation.

     

Cannabimimetic activities of cumyl carboxamide-type synthetic cannabinoids

1-Pentyl-N-(2-phenylpropan-2-yl)-1H-indazole-3-carboxamide (CUMYL-PINACA) is a carboxamide-type synthetic cannabinoid comprising a cumylamine moiety. Recently, the detection of CUMYL-PINACA and some analogs in illicit drug products has been reported by the European Monitoring Centre for Drugs and Drug Addiction. In this study, we synthesized seven cumyl carboxamide-type synthetic cannabinoids (CUMYL-PINACA, CUMYL-5F-PINACA, CUMYL-PICA, CUMYL-5F-PICA, CUMYL-THPINACA, CUMYL-BICA, and CUMYL-5F-P7AICA) and evaluated their activities as CB₁ and CB₂ receptor agonists. We also showed the analytical characterization of these compounds using gas chromatography–electron ionization-mass spectrometry. All of the evaluated compounds exhibited moderate to strong activities as agonists acting on both CB₁ and CB₂ receptors with EC₅₀ values in the range of 8.1 × 10^?10–7.8 × 10^?7 mol/L for CB₁ and from 2.5 × 10^?10 to 9.1 × 10^?6 mol/L for CB₂. The EC₅₀ data presented will be helpful to understand the effects of these compounds in the forensic cases. Furthermore, other new cumyl carboxamide-type synthetic cannabinoids, which will be potentially distributed in the future, will probably have the activities as agonists acting on both CB₁ and CB₂ receptors.     

Identification and quantitation of two cannabimimetic phenylacetylindoles JWH-251 and JWH-250, and four cannabimimetic naphthoylindoles JWH-081, JWH-015, JWH-200, and JWH-073 as designer drugs in illegal products

Six cannabimimetic indoles have been identified as adulterants in herbal or chemical products being sold illegally in Japan, with four of the compounds being new as adulterants to our knowledge. The identifications were based on analyses using gas chromatography–mass spectrometry, liquid chromatography–mass spectrometry, high-resolution mass spectrometry, and nuclear magnetic resonance spectroscopy. The first two compounds were identified as phenylacetyl indoles JWH-251 (2-(2-methylphenyl)-1-(1-pentyl-1H-indol-3-yl)ethanone; 1) and its demethyl-methoxylated analog JWH-250 (2-(2-methoxyphenyl)-1-(1-pentyl-1H-indol-3-yl)ethanone; 2). Compound 2 was identical to that found as an adulterant in the UK and in Germany in 2009. The third compound was naphthoylindole JWH-081 (1-(4-methoxynaphthalenyl)-(1-pentyl-1H-indol-3-yl)methanone; 3), and the fourth was JWH-073 (1-naphthalenyl(1-butyl-1H-indol-3-yl)methanone; 4), which had been identified as an adulterant in our previous study. Two additional compounds were JWH-015 (1-naphthalenyl(2-methyl-1-propyl-1H-indol-3-yl)methanone; 5) and JWH-200 (1-naphthalenyl(1-(2-(4-morpholinyl)ethyl)-1H-indol-3-yl)methanone; 6). Compounds 1–4 and 6 were reported to be synthetic cannabinoids with selective affinity for cannabinoid CB₁ receptors, while compound 5 was reported to be a selective CB₂ receptor agonist causing immunosuppressive effects without psychotropic affects. One product contained both CB₁ and CB₂ receptor agonists in our collection. Quantitative analyses of the six cannabimimetic compounds in 20 products revealed that there was large variation in concentrations of the detected compounds among products; for herbal cutting products, the total amounts of these cannabinoids ranged from 26 to 100 mg.     

Synthesis and preliminary evaluation of [18F]-labeled 2-oxoquinoline derivatives for PET imaging of cannabinoid CB2 receptor

IntroductionThe cannabinoid receptor type 2 (CB₂) is an important target for development of drugs and imaging agents for diseases, such as neuroinflammation, neurodegeneration and cancer. Recently, we reported synthesis and results of in vitro receptor binding of a focused library of fluorinated 2-oxoquinoline derivatives as CB₂ receptor ligands. Some of the compounds demonstrated to be good CB₂-specific ligands with Ki values in the nanomolar to subnanomolar concentrations; therefore, we pursued the development of their ¹⁸F-labeled analogues that should be useful for positron emission tomography (PET) imaging of CB₂ receptor expression. Here, we report the radiosynthesis of two ¹⁸F-labeled 2-oxoquinoline derivatives and the preliminary in vitro and ex vivo evaluation of one compound as a CB₂-specific radioligand.Methods4-[¹⁸F]fluorobenzyl amine [¹⁸F]-3 was prepared by radiofluorination of 4-cyano-N,N,N-trimethylanilinium triflate salt followed by reduction with LiAlH₄ and then coupled with acid chlorides 11 and 12 to afford [¹⁸F]-13 and [¹⁸F]-14. In vitro CB₂ receptor binding assay was performed using U87 cells transduced with CB₂ and CB₁ receptor. Ex vivo autoradiography was performed with [¹⁸F]-14 on spleen and on CB₂- and CB₁-expressing and wild-type U87 subcutaneous tumors grown in mice.ResultsThe radiochemical yields of [¹⁸F]-13 and [¹⁸F]-14 were 10%–15.0% with an average of 12% (n=10); radiochemical purity was >99% with specific activity 1200 mCi/μmol. The dissociation constant Kd for [¹⁸F]-14 was 3.4 nM. Ex vivo autoradiography showed accumulation of [¹⁸F]-14 in the CB₂-expressing tumor.ConclusionTwo new [¹⁸F]-labeled CB₂ ligands have been synthesized. Compound [¹⁸F]-14 appears to be a potential PET imaging agent for the assessment of CB₂ receptor expression; however, poor solubility restrain its use in vivo.     

In vitro metabolism of new synthetic cannabinoid SDB-006 in human hepatocytes by high-resolution mass spectrometry

The drug abuse epidemic within the United States remains one of the nation’s most serious social challenges, especially among adolescents and young adults. Novel psychoactive substances continuously emerge into the illicit drugs-of-abuse market to evade legislation. In 2013, SDB-006 was detected as a novel synthetic cannabinoid (SC) with high binding affinity to CB₁ (EC₅₀ = 19 nM) and CB₂ (EC₅₀ = 134 nM). Unfortunately, no human metabolism data for SDB-006 are currently available, making it challenging to confirm intake, since all previously investigated SCs were extensively metabolized. The present study aims to recommend appropriate marker metabolites for documenting SDB-006 consumption by investigating its metabolism in human hepatocytes. For metabolite profiling, 10 µM of SDB-006 was incubated in human hepatocytes for 3 h. Metabolite identification in hepatocyte samples was accomplished with high-resolution mass spectrometry via information-dependent data acquisition. Results revealed that SDB-006 was highly metabolized in human hepatocytes. A total of 20 metabolites were characterized, generated mainly from hydroxylation and glucuronidation. Hydroxylation occurred primarily on several positions of the pentyl chain. N-Dealkylation was the other major pathway, including depentylation and debenzylation. Based on our data, we propose 4′-keto-SDB-006 (M19) and pentyl-OH-SDB-006 (M15) as optimal marker metabolites for documenting SDB-006 intake.     

Synthesis and biological evaluation of carbon-11- and fluorine-18-labeled 2-oxoquinoline derivatives for type 2 cannabinoid receptor positron emission tomography imaging

IntroductionThe type 2 cannabinoid (CB₂) receptor is part of the endocannabinoid system and has been suggested as a mediator of several central and peripheral inflammatory processes. Imaging of the CB₂ receptor has been unsuccessful so far. We synthesized and evaluated a carbon-11- and a fluorine-18-labeled 2-oxoquinoline derivative as new PET tracers with high specificity and affinity for the CB₂ receptor.MethodsTwo 2-oxoquinoline derivatives were synthesized and radiolabeled with either carbon-11 or fluorine-18. Their affinity and selectivity for the human CB₂ receptor were determined. Biological evaluation was done by biodistribution, radiometabolite and autoradiography studies in mice.ResultsIn vitro studies showed that both compounds are high affinity CB₂-specific inverse agonists. Biodistribution study of the tracers in mice showed a high in vivo initial brain uptake and fast brain washout, in accordance with the low CB₂ receptor expression levels in normal brain. A persistently high in vivo binding to the spleen was observed, which was inhibited by pretreatment with two structurally unrelated CB₂ selective inverse agonists. In vitro autoradiography studies with the radioligands confirmed CB₂-specific binding to the mouse spleen.ConclusionWe synthesized two novel CB₂ receptor PET tracers that show high affinity/selectivity for CB₂ receptors. Both tracers show favourable characteristics as radioligands for central and peripheral in vivo visualization of the CB₂ receptor and are promising candidates for primate and human CB₂ PET imaging.     

Long-term effects of sprint interval training on expression of cardiac genes involved in energy efficiency

Purpose

The real mechanisms of intensive exercise training-induced energy efficiency have not yet been well examined. Therefore, the aim of the present study was to investigate the effects of sprint interval training (SIT) on gene expression of uncoupling proteins (UCPs) and endothelial nitric oxide synthase (eNOS).

Methods

For this purpose, 16 Albino Wistar rats (250–300 g) were randomly divided into equal groups of control and sprint training. The animals run on treadmill for 10 weeks, 5 days per week at intensity corresponding to 90–95% maximal oxygen consumption. The gene expression of UCP2, UCP3 and eNOS was analyzed by RT-PCR method in hearts. The data were analyzed by independent samples T test at P?<?0.05 level.

Results

Sprint interval training significantly decreased mRNA expression of UCP2 (t₁₄?=?4.818, P?=?0.001) and UCP3 (t₁₄?=?4.620, P?=?0.001) in cardiac muscle of rats. In contrast, mRNA expression of eNOS in cardiac muscle significantly increased following sprint interval training (t₁₄?=?7.967, P?=?0.001).

Conclusion

This study elucidates that SIT through reduction in gene expression of uncoupling proteins can improve energy efficiency. But, more studies are needed to confirm this hypothesis.

     

MDCTA volumetric analysis for the quantification and grading of acute non-cerebral,non-gastrointestinal hemorrhage: a feasibility study

Purpose

To assess the feasibility of multi-detector computed tomography angiography (MDCTA) volumetry for the quantification and grading of acute non-cerebral, non-gastrointestinal bleeding.

Methods

This retrospective, single-center study investigated consecutive patients with MDCTA positive for active non-cerebral, non-gastrointestinal bleeding, between January 2020 and June 2020. Outcome measures were the quantification of active extravasation at the arterial and parenchymal phase using volumetry measurements, the calculation of active bleeding rate and bleeding grading, 30-day mortality rate, identification of independent predictors of mortality and correlation between volumetric analysis, various clinical features, and the decision to proceed with an intervention.

Results

In total 30 patients (17 females; 56.6%; mean age 70.0?±?16.0 years) were analyzed. Volumetric analysis was feasible in all cases resulting in excellent inter-observer variability (interclass correlation coefficient 0.999 for arterial and 0.919 for venous volume measurements). Mean volume of contrast extravasation was 1.06?±?1.09 ml and 3.07?±?2.48 ml at the arterial and parenchymal phases, respectively. Mean bleeding rate was 6.95?±?7.82 ml/min. High bleeding volume at arterial phase (grade 4 bleeding) was the only independent predictor of 30-day mortality (HR 1383.58; p?=?0.042). There was a positive correlation between bleeding volume at arterial phase (r_s?=?0.340; p?=?0.033) and arterial bleeding rate (r_s?=?0.381; p?=?0.019) with the decision to proceed with an intervention. Bleeding volume of 0.6 ml was the cutoff value for the prediction of intervention (sensitivity 96.3%; specificity 66.7%).

Conclusions

MDCTA volumetric analysis for the quantification and grading of acute hemorrhage was feasible with excellent inter-observer agreement. The proposed bleeding grading system could optimize decision making and predict clinical outcomes.

     

Relationship between vastus lateralis muscle ultrasound echography,knee extensors rate of torque development,and jump height in professional soccer athletes

Purpose

This study aimed to investigate the relationship between knee extensors maximum voluntary contraction (MVC) torque and rate of torque development (RTD) with jump performance and with echography intensity (EI) from the vastus lateralis muscle (VL).

Methods

We assessed the MVC torque and RTD from knee extensors by a standard isokinetic device from 16 male professional soccer players (25.5?±?3.9 years). Counter-movement jump (CMJ) and squat jump (SJ) height were calculated from their flight times. EI was determined from VL ultrasound images’ grayscale histogram acquired in the middle of the tight. Correlation between variables was investigated by the Pearson correlation coefficient.

Results

We observed a VL EI of 26.4?±?7.8 a.u., and the SJ and CMJ heights were 36.0?±?3.0 and 36.2?±?4.1 cm, respectively. There was a significant correlation between the MVC torque and absolute late-phase RTD (r?=?0.67 and 0.76—RTD₂₀₀ and RTD₂₅₀, respectively), between the CMJ height and absolute RTD₅₀ (r?=?0.50), and between the normalized early-phase RTD and SJ height (r?=?0.53–0.60—RTD₅₀ and RTD₁₅₀). Additionally, normalized RTD₂₀₀ (r?=?? 0.51) and RTD₂₅₀ (r?=?? 0.56) were negatively correlated with EI.

Conclusions

Such results suggest that athletes with the ability to produce torque explosively in the very beginning (between 50 and 150 ms) of knee extension can jump higher. Also, the muscle quality assessed by the VL IE seems to be more important to maximum strength and later periods of torque rise (>?200 ms).

     

Diagnostic performance of 2D-shear wave elastography in the diagnosis of breast cancer: a clinical appraisal of cutoff values

Purpose

To assess the role of 2D-shear wave elastography (2D-SWE) in differentiating benign from malignant focal breast lesions (FBLs), providing new vendor-specific cutoff values.

Methods

158 FBLs (size: 3.5–50 mm) detected in 151 women (age: 21–87 years) were prospectively evaluated by means 2D-SWE. For each lesion, an expert radiologist assessed US BI-RADS category and calculated the following four 2D-SWE parameters: (1) elasticity maximum (E_max); (2) mean elasticity (E_mean); (3) minimum elasticity (E_min); (4) elasticity ratio (E_ratio). US-guided core-biopsy was considered as standard of reference for all the FBLs classified as BI-RADS 4 or 5. For each 2D-SWE parameter, the optimal cutoff value for a diagnostic test was calculated using the Youden method. Diagnostic performance of the US BI-RADS and 2D-SWE parameters was calculated accordingly.

Results

83/158 (52.5%) FBLs were benign and 75/158 (47.5%) were malignant. Statistically significant higher stiffness values were observed in malignant FBLs for all 2D-SWE parameters than in benign ones (p?<?0.001). 2D-SWE cutoff values were 82.6 kPa, 66.0 kPa and 53.6 kPa, respectively, for E_max, E_mean, E_min and 330.8% for E_ratio. The 2D-SWE parameter showing the best diagnostic accuracy was E_max (85.44%). Considering US BI-RADS 3 (n?=?60) and 4a (n?=?32) FBLs, E_max and E_mean showed the best diagnostic accuracy (85.87% for both), without a statistically significant decrease in sensitivity (p?=?0.7003 and p?=?1, respectively).

Conclusion

Our study provides new vendor-specific cutoff values for 2D-SWE, suggesting its possible clinical use in the adjunctive assessment of category US-BI-RADS 3 and 4a breast masses.

     

Type 1 cannabinoid receptor mapping with [18F]MK-9470 PET in the rat brain after quinolinic acid lesion: a comparison to dopamine receptors and glucose metabolism

Purpose

Several lines of evidence imply early alterations in metabolic, dopaminergic and endocannabinoid neurotransmission in Huntington’s disease (HD). Using [¹⁸F]MK-9470 and small animal PET, we investigated cerebral changes in type 1 cannabinoid (CB₁) receptor binding in the quinolinic acid (QA) rat model of HD in relation to glucose metabolism, dopamine D₂ receptor availability and amphetamine-induced turning behaviour.

Methods

Twenty-one Wistar rats (11 QA and 10 shams) were investigated. Small animal PET acquisitions were conducted on a Focus 220 with approximately 18 MBq of [¹⁸F]MK-9470, [¹⁸F]FDG and [¹¹C]raclopride. Relative glucose metabolism and parametric CB₁ receptor and D₂ binding images were anatomically standardized to Paxinos space and analysed voxel-wise using Statistical Parametric Mapping (SPM2).

Results

In the QA model, [¹⁸F]MK-9470 uptake, glucose metabolism and D₂ receptor binding were reduced in the ipsilateral caudate-putamen by 7, 35 and 77%, respectively (all p?<?2.10^?5), while an increase for these markers was observed on the contralateral side (>5%, all p?<?7.10^?4). [¹⁸F]MK-9470 binding was also increased in the cerebellum (p?=?2.10^?5), where it was inversely correlated to the number of ipsiversive turnings (p?=?7.10^?6), suggesting that CB₁ receptor upregulation in the cerebellum is related to a better functional outcome. Additionally, glucose metabolism was relatively increased in the contralateral hippocampus, thalamus and sensorimotor cortex (p?=?1.10^?6).

Conclusion

These data point to in vivo changes in endocannabinoid transmission, specifically for CB₁ receptors in the QA model, with involvement of the caudate-putamen, but also distant regions of the motor circuitry, including the cerebellum. These data also indicate the occurrence of functional plasticity on metabolism, D₂ and CB₁ neurotransmission in the contralateral hemisphere.     

Evaluation of carboxamide-type synthetic cannabinoids as CB<Subscript>1</Subscript>/CB<Subscript>2</Subscript> receptor agonists: difference between the enantiomers

Recently, carboxamide-type synthetic cannabinoids have been distributed globally as new psychoactive substances (NPS). Some of these compounds possess asymmetric carbon, which is derived from an amide moiety composed of amino acid derivatives (i.e., amides or esters of amino acids). In this study, we synthesized both enantiomers of synthetic cannabinoids, N-(1-amino-3-methyl-1-oxobutan-2-yl)-1-(2-fluorobenzyl)-1H-indazole-3-carboxamide (AB-FUBINACA 2-fluorobenzyl isomer), N-(1-amino-1-oxo-3-phenylpropan-2-yl)-1-(cyclohexylmethyl)-1H-indazole-3-carboxamide (APP-CHMINACA), ethyl [1-(5-fluoropentyl)-1H-indazole-3-carbonyl]valinate (5F-EMB-PINACA), ethyl [1-(4-fluorobenzyl)-1H-indazole-3-carbonyl]valinate (EMB-FUBINACA), and methyl 2-[1-(4-fluorobenzyl)-1H-indole-3-carboxamido]-3,3-dimethylbutanoate (MDMB-FUBICA), which were reported as NPS found in Europe from 2014 to 2015, to evaluate their activities as CB₁/CB₂ receptor agonists. With the exception of (R) MDMB-FUBICA, all of the tested enantiomers were assumed to be agonists of both CB₁ and CB₂ receptors, and the EC₅₀ values of the (S)-enantiomers for the CB₁ receptors were about five times lower than those of (R)-enantiomers. (R) MDMB-FUBICA was shown to function as an agonist of the CB₂ receptor, but lacks CB₁ receptor activity. To the best of our knowledge, this is the first report to show that the (R)-enantiomers of the carboxamide-type synthetic cannabinoids have the potency to activate CB₁ and CB₂ receptors. The findings presented here shed light on the pharmacological properties of these carboxamide-type synthetic cannabinoids in forensic cases.     

Foam rolling is an effective recovery tool in trained distance runners

Purpose

Many endurance athletes use foam rolling (FR) to decrease muscle soreness, but it is unclear whether FR effectively treats soreness in this population. Moreover, the effects of FR in highly trained runners are unknown. The aim of this study was to use downhill running (DHR) to induce muscle soreness in runners and to determine the influence of FR on soreness and running performance when compared to sham compression tights.

Methods

Participants performed a running economy (RE) test at 75% of 5-km race speed and a 3-km time trial (TT). In a crossover design, subjects then completed DHR followed by either a FR protocol or wearing sham compression tights. Two days post-DHR, subjects repeated the RE and TT tests. Crossover visits occurred 2–4 weeks later. During RE tests, VO₂ and rating of perceived exertion (RPE) were recorded. Passive and active soreness were measured on a scale of 0 (no soreness) to 10 (extreme soreness).

Results

Eight runners (aged 31?±?7 years; four females; VO_2peak 57?±?7 ml kg^?1 min^?1) completed the study. Both treatment conditions experienced passive (p?=?0.026) and active soreness (p = 0.012) induced by DHR. Active soreness 2 days post-DHR was significantly lower after FR than after sham compression tights (p?=?0.025). With tights, there was a trend for an increased RPE compared to pre-DHR (p?=?0.056).

Conclusions

Foam rolling decreases leg soreness in well-trained runners and attenuates soreness-related increases in perceived exertion during sub-maximal running.

     

Identification of (1-pentylindol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone and its 5-pentyl fluorinated analog in herbal incense seized for drug trafficking

Four herbal incense products were seized from suspected drug abusers in Korea. The major ingredients in the herbal incense samples were purified, and their structures were elucidated using gas chromatography–electron ionization–mass spectrometry (GC–EI–MS), liquid chromatography–time-of-flight–mass spectrometry (LC–TOF–MS), and 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. As a result, ingredients in the herbal incense were identified as (1-pentylindol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone and its 5-pentyl fluorinated analog [1-(5-fluoropentyl)indol-3-yl]-(2,2,3,3-tetramethylcyclopropyl)methanone. The former is being sold via the Internet as a "research chemical" named UR-144, and the latter is sold as 5F-UR-144. UR-144 is a selective full agonist of CB₂ cannabinoid receptor, and was first developed by Abbott Laboratories as an analgesic. It exhibits analgesic activity against both neuropathic and inflammatory pain associated mainly with the CB₂ receptor, but shows less psychotropic effects associated with the CB₁ receptor. Fluorination of the N-pentyl side chain of cannabimimetic compounds increases their cannabinoid receptor affinity such as with AM-2201, which shows both increased analgesic and psychotropic effects simultaneously. UR-144 and 5F-UR-144 can be classified as research chemicals based on their analgesic effects, but in practice are abused as psychotropic agents and can cause unexpected toxic effects. Thus, the trade and diversion of these chemicals should be monitored carefully for possible abuse. To our knowledge, this is the first report disclosing cyclopropylcarbonylindoles in herbal products.     

Good functional outcomes in patient’s age > 40 years after anterior cruciate ligament reconstruction with hamstring tendon graft: a retrospective study

Background

Many surgeons are still hesitant to do arthroscopic anterior cruciate ligament reconstruction (ACL) in patients above 40 years old.

Purpose

The effect of ACL reconstruction on functional outcomes in patients above 40 years of age.

Materials and methods

Patients of age?>?40 years who underwent ACL reconstruction were enrolled in the study. Functional outcomes (Lysholm, WOMAC score, and Tegner activity scale), pain (VAS score), and knee stability (KT-1000) were assessed at final follow-up and compared with pre-operative scores. All patients were followed up for a minimum of 2 years.

Results

A total of 112 patients were included in the study. Eighty-eight were males and 24 were females. The mean age at the time of surgery was 44.8?±?5.6 years (40–63 years). The mean Lysholm score was improved from 64.1?±?11.7 to 87.2?±?5.9 (p?=?0.0001). Similarly, the WOMAC score was also improved from 8.8?±?6.9 to 2.3?±?1.9 (p?=?0.0001). The mean side-to-side difference (KT-1000) also improved from 6.65?±?2.25 mm to 2.32?±?1.45 mm (p?=?0.0001). More than 70% of the patients had excellent or good outcomes. Median pre-injury Tegner activity scale was 5 (3–8) and the median post-ACLR Tegner activity scale was 5 (3–7; p?=?1).

Conclusion

ACL reconstruction in patients?>?40 years of age results in good functional outcomes and knee stability.

     

Water-based resistance training program with isolated concentric action improves physical functional capacity and muscular strength in older women

Background

Resistance training has proven to be an excellent method for counteracting aging physical dysfunctions. However, its application in the liquid environment is not yet fully elucidated.

Aim

To investigate the effects of water-based resistance training (WBRT) with the concentric phase performed as fast as possible, compared to conventional resistance training (CRT), on physical functional capacity, muscle strength, and body composition in older women.

Methods

Thirteen healthy older women participated in the WBRT and 11 in the CRT. Estimation statistics focused on the effect size of the experiment/intervention were used. We also analyzed the intervention effect based on the percentage delta between WBRT and CRT.

Results

The WBRT group showed a negative large effect (d?=?? 0.922; p?=?0.0274) for the timed up and go, and a large effect for chair rise in 30″ and the elbow flex test (d?=?1.58; p?=?0.0012; d?=?2.8; p?=?0.01) respectively. Intervention comparisons based on the delta percentage between WBRT and CRT presented an intermediate effect (d?=?0.606; p?=?0.157) for the stair climb, a large effect (d?=?0.988; p?=?0.0282) for the timed up and go, and a large negative effect [d?=?? 1.32 (90.0% CI ? 1.92, ? 0.646); p?=?0.0038] for the elbow flex test. Concentric extensor-flexor peak torque (60°/s) showed an intermediate effect (d?=?0.749; p?=?0.0876; d?=?0.65; p?=?0.122 respectively). Body fat (%) demonstrated an intermediate effect (d?=?0.523; p?=?0.234).

Conclusion

WBRT with the concentric phase performed as fast as possible was able to improve physical functional capacity and maximal knee extension strength of older women.

     

Hyperreflexia induced by XLR-11 smoke is caused by the pyrolytic degradant

Purpose

Some of the synthetic cannabinoids, often found in recreational drugs of the herbal form, reportedly induce a generalized seizure in drug abusers immediately after smoking. However, it is still unclear what elicits the sensorimotor responses, particularly in the case of hyperreflexia or excitatory behavior during the synthetic cannabinoid exposure. The purpose of this study was to explore the mechanism underlying the hyperreflexia induced by smoke intoxication of XLR-11 [(1-(5-fluoropentyl)-1H-indol-3-yl)(2,2,3,3-tetramethylcyclopropyl)methanone].

Methods

Locomotor activity and body temperature of mice were measured using an implanted Nano-Tag device. The intensity of catalepsy was determined by the bar test. The extracellular dopamine levels in the nucleus accumbens and glutamate levels in the hippocampus were measured by in vivo microdialysis using electrochemical detector-coupled high-performance liquid chromatography and by in vivo enzyme-based biosensor method, respectively.

Results

Mice exposed to the smoke of XLR-11 exhibited hyperreflexia at the very early phase, followed by hypothermia and catalepsy. The XLR-11 smoke contained XLR-11 and XLR-11 degradant at a ratio of approximately 1:25. Mice treated intraperitoneally with XLR-11 degradant at a dose comparable to the smoke inhalation experiment showed a hyperreflexic effect immediately after the treatment, but XLR-11 showed no such effect. The effects of XLR-11 degradant were significantly suppressed by pretreatment with AM-251, a CB₁ receptor antagonist. Extracellular dopamine and glutamate levels showed no evidence of involvement in the XLR-11 degradant-induced hyperreflexia; on the other hand gabapentin, a GABAergic antiepileptic, significantly suppressed the enhanced locomotor activity.

Conclusions

The hyperreflexic effect of XLR-11 degradant is mediated by the CB₁ receptor and possibly by GABAergic function.