Recently, an increasing number of pharmacists had to supply medicinal products based on L. (Cannabaceae), prescribed by physicians to individual patients. Cannabis olive oil preparation is the first choice as a concentrated extract of cannabinoids, even though standardized operative conditions for obtaining it are still not available. In this work, the impact of temperature and extraction time on the concentration of active principles was studied to harmonize the different compounding methods, optimize the extraction process, and reduce the variability among preparations. Moreover, starting from the cannabis inflorescence, the effect of temperature on tetrahydrocannabinolic acid decarboxylation was evaluated. For the analysis, a GC/MS method, as suggested by the Italian Ministry of Health, and a GC/flame ionization detection method were developed, validated, and compared.

Gonadal function was elevated in 80 male heroin and/or methadone addicts by measuring basal plasma levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and testosterone. In 41 subjects semen analyses were also undertaken. Three groups were distinguished consisting of 15 heroin addicts, 42 undergoing methadone treatment but continuing to take heroin, and 23 taking only methadone. All patients had normal plasma levels of FSH, LH and testosterone. Prolactin levels were normal in all subjects except for the 15 heroin addicts, in whom they were significantly higher than in controls (P less than 0.025). Semen analyses from all of the heroin addicts and from the dual heroin-methadone users were abnormal, whereas only 10 out of 22 (45%) of the methadone takers were pathological. In all cases asthenospermia was one of the abnormalities (100%). Twenty-four per cent also showed teratospermia and hypospermia and 17% showed oligozoospermia. Such seminal pathology, especially of forward motility, even in combination with normal hormone levels, might be an early indication of heroin toxicity to the male reproductive tract.

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Gut microbiota metabolization of dietary choline may promote atherosclerosis through trimethylamine (TMA), which is rapidly absorbed and converted in the liver to proatherogenic trimethylamine-N-oxide (TMAO). The aim of this study was to verify whether TMAO urinary levels may be associated with the fecal relative abundance of specific bacterial taxa and the bacterial choline TMA-lyase gene cutC. The analysis of sequences available in GenBank grouped the cutC gene into two main clusters, cut-Dd and cut-Kp. A quantitative real-time polymerase chain reaction (qPCR) protocol was developed to quantify cutC and was used with DNA isolated from three fecal samples collected weekly over the course of three consecutive weeks from 16 healthy adults. The same DNA was used for 16S rRNA gene profiling. Concomitantly, urine was used to quantify TMAO by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). All samples were positive for cutC and TMAO. Correlation analysis showed that the cut-Kp gene cluster was significantly associated with Enterobacteriaceae. Linear mixed models revealed that urinary TMAO levels may be predicted by fecal cut-Kp and by 23 operational taxonomic units (OTUs). Most of the OTUs significantly associated with TMAO were also significantly associated with cut-Kp, confirming the possible relationship between these two factors. In conclusion, this preliminary method-development study suggests the existence of a relationship between TMAO excreted in urine, specific fecal bacterial OTUs, and a cutC subgroup ascribable to the choline-TMA conversion enzymes of Enterobacteriaceae.