Comprehending and improving cannabis specialized metabolism in the systems biology era.

Cannabis sativa is a source of food, fiber and specialized metabolites such as cannabinoids, with psychoactive and pharmacological effects. Due to its expanding and increasingly-accepted use in medicine, cannabis cultivation is acquiring more importance and less social stigma. Humans initiated different domestication episodes whose later spread gave rise to a plethora of landrace cultivars. At present, breeders cross germplasms from different gene pools depending on their specific use. The fiber (hemp) and drug (marijuana) types of C. sativa differ in their cannabinoid chemical composition phenotype (chemotype) and also in the accumulation of terpenoid compounds that constitute a strain's particular flavor and scent. Cannabinoids are isoprenylated polyketides among which cannabidiolic acid (CBDA) and (-)-trans-Δ9-tetrahydrocannabinol acid (THCA) have been well-documented for their many effects on humans. Here, we review the most studied specialized metabolic pathways in C. sativa, showing how terpenes and cannabinoids share both part of the isoprenoid pathway and the same biosynthetic compartmentalization (i.e. glandular trichomes of leaves and flowers). We enlist the several studies that have deciphered these pathways in this species including physical and genetic maps, QTL analyses and localization and enzymatic studies of cannabinoid and terpene synthases. In addition, new comparative modeling of cannabinoid synthases and phylogenetic trees are presented. We describe the genome sequencing initiatives of several accessions with the concomitant generation of next-generation genome maps and transcriptomic data. Very recently, proteomic characterizations and systems biology approaches such as those applying network theory or the integration of multi-omics data have increased the knowledge on gene function, enzyme diversity and metabolite content in C. sativa. In this revision we drift through the history, present and future of cannabis research and on how second- and third-generation sequencing technologies are bringing light to the field of cannabis specialized metabolism. We also discuss different biotechnological approaches for producing cannabinoids in engineered microorganisms.

Síndrome poliglandular autoinmunitario tipo II (síndrome de Schmidt): enfermedad de Addison y tiroiditis de Hashimoto asociado con vitíligo / Polyglandular autoimmune syndrome type II (Schmidt syndrome): Addison's disease and Hashimoto's thyroiditis associated to vitiligo

Resumen Los síndromes poliglandulares autoinmunitarios son afecciones poco frecuentes que se distinguen por la coexistencia de al menos dos enfermedades glandulares autoinmunitarias. Se clasifican en tipo I (o juvenil) y tipos II y III (o del adulto). El tipo II o síndrome de Schmidt se caracteriza por enfermedad de Addison, enfermedad tiroidea autoinmunitaria o diabetes mellitus tipo 1 que pueden vincularse con otras alteraciones de naturaleza autoinmunitaria, como vitíli go, hepatitis autoinmunitaria, miastenia gravis, anemia perniciosa, enfermedad celiaca y alopecia areata, entre otras. Se comunica el caso de una paciente de 61 años de edad con vitíligo a quien se le diagnosticó enfermedad de Addison y tiroiditis de Hashimoto (síndrome de Schmidt).

Abstract Autoimmune polyglandular syndromes are rare conditions characterized by the coexistence of at least two autoimmune glandular diseases. They can be classified in type I (or juvenile) and type II and III (or adult). Type 2 or Schmidt's syndrome is characterized by Addison's disease, autoimmune thyroid disease and/or type 1 diabetes mellitus and may be associated with other disorders of autoimmune nature, such as vitiligo, autoimmune hepatitis, myasthenia gravis, pernicious anemia, celiac disease or alopecia areata, among others. We communicate the case of a 61 year-old woman with vitiligo diagnosed with Addison's disease and Hashimoto's thyroiditis (Schmidt's syndrome).

Interleukin-1ß increases neuronal death in the hippocampal dentate gyrus associated with status epilepticus in the developing rat. / La interleucina-1ß aumenta la muerte neuronal en el giro dentado del hipocampo asociada al estado epiléptico en la rata en desarrollo.

BACKGROUND: Interleukin-1ß (IL-1ß) increases necrotic neuronal cell death in the CA1 area after induced status epilepticus (SE) in developing rats. However, it remains uncertain whether IL-1ß has a similar effect on the hippocampal dentate gyrus (DG). In this study, we analysed the effects of IL-1ß on 14-day-old Wistar rats experiencing DG neuronal death induced by SE. METHODS: SE was induced with lithium-pilocarpine. Six hours after SE onset, a group of pups was injected with IL-1ß (at 0, 0.3, 3, 30, or 300ng/µL) in the right ventricle; another group was injected with IL-1ß receptor (IL-1R1) antagonist (IL-1Ra, at 30ng/µL) of IL-1RI antagonist (IL-1Ra) alone, and additional group with 30ng/µL of IL-1Ra plus 3ng/µL of IL-1ß. Twenty-four hours after SE onset, neuronal cell death in the dentate gyrus of the dorsal hippocampus was assessed using haematoxylin-eosin staining. Dead cells showed eosinophilic cytoplasm and condensed and fragmented nuclei. RESULTS: We observed an increased number of eosinophilic cells in the hippocampal DG ipsilateral to the site of injection of 3ng/µL and 300ng/µL of IL-1ß in comparison with the vehicle group. A similar effect was observed in the hippocampal DG contralateral to the site of injection of 3ng/µL of IL-1ß. Administration of both of IL-1ß and IL-1Ra failed to prevent an increase in the number of eosinophilic cells. CONCLUSION: Our data suggest that IL-1ß increases apoptotic neuronal cell death caused by SE in the hippocampal GD, which is a mechanism independent of IL-1RI activation.

Pharmacokinetics and lung and muscle concentrations of tulathromycin following subcutaneous administration in white-tailed deer (Odocoileus virginianus).

Respiratory tract infections are common in farmed North American white-tailed deer (Odocoileus virginianus). Tulathromycin is approved for use in cattle but not deer but is often employed to treat deer. The pharmacokinetic properties and lung and muscle concentrations of tulathromycin in white-tailed deer were investigated. Tulathromycin was administered to 10 deer, and then, serum, lung, and muscle tulathromycin concentrations were measured using liquid chromatography-mass spectrometry (LC-MS). The mean maximal serum tulathromycin concentration in deer was 359 ng/mL at 1.3 h postinjection. The mean area under the serum concentration-time curve, apparent volume of distribution, apparent clearance, and half-life was 4883 ng·h/mL, 208 L/kg, 0.5 L/h/kg, and 281 h (11.7 days), respectively. The maximal tulathromycin concentration in lung and muscle homogenate from a single animal was 4657 ng/g (14 days) and 2264 ng/g (7 days), respectively. The minimum concentrations in lung and muscle were 39.4 ng/g (56 days) and 9.1 ng/g (56 days), respectively. Based on similarity in maximal serum concentrations between deer and cattle and high lung concentrations in deer, we suggest the recommended cattle dosage is effective in deer. Tissue concentrations persisted for 56 days, suggesting a need for longer withdrawal times in deer than cattle. Further tissue distribution and depletion studies are necessary to understand tulathromycin persistence in deer tissue; clinical efficacy studies are needed to confirm the appropriate dosage regimen in deer.

Pharmacokinetics of tulathromycin after subcutaneous injection in North American bison (Bison bison).

Tulathromycin is approved for the treatment of respiratory disease in cattle and swine. It is intended for long-acting, single-dose injection therapy (Draxxin), making it particularly desirable for use in bison due to the difficulty in handling and ease of creating stress in these animals. The pharmacokinetic properties of tulathromycin in bison were investigated. Ten wood bison received a single 2.5 mg/kg subcutaneous injection of Draxxin. Serum concentrations were measured by liquid chromatography-mass spectrometry (LC-MS) detection. Tulathromycin demonstrated early maximal serum concentrations, extensive distribution, and slow elimination characteristics. The mean maximum serum concentration (Cmax) was 195 ng/mL at 1.04 h (tmax) postinjection. The mean area under the serum concentration-time curve, extrapolated to infinity (AUC0-inf ), was 9341 ng · h/mL. The mean apparent volume of distribution (Vd /F) and clearance (Cls/F) was 111 L/kg and 0.4 L/h/kg, respectively, and the mean half-life (t1/2) was 214 h (8.9 days). Compared to values for cattle, Cmax and AUC0-inf were lower in bison, while the Vd /F was larger and the t1/2 longer. Tissue distribution and clinical efficacy studies in bison are needed to confirm the purported extensive distribution of tulathromycin into lung tissue and to determine whether a 2.5 mg/kg subcutaneous dosage is adequate for bison.

Isolation of WDR and bHLH genes related to flavonoid synthesis in grapevine (Vitis vinifera L.).

Anthocyanins and tannins are two of the most abundant flavonoids found in grapevine, and their synthesis is derived from the phenylpropanoid pathway. As described for model species such as Arabidopsis thaliana, maize and petunia, the end-point branches of this pathway are tightly regulated by the combinatorial interaction of three families of regulatory factors; MYB, bHLH (also known as MYC) and WDR proteins. Among these, only MYB genes have been previously identified in grapes. Here, we report the isolation of the first members from the WDR and bHLH families found in Vitis vinifera, named WDR1, WDR2 and MYCA1. WDR1 contributed positively to the accumulation of anthocyanins when it was overexpressed in A. thaliana, although it was not possible to determine the function of WDR2 by ectopic expression. The sub-cellular localizations of WDR1 and MYCA1 were observed by means of GFP-fusion proteins, indicating both cytoplasm and nuclear localization, in contrast to the localization of a MYB factor exclusively in the nucleus. The expression patterns of these genes were quantified in coloured reproductive organs throughout development, and correlated with anthocyanin accumulation and the expression profiles of the flavonoid-related MYBA1-2, UFGT, and ANR genes. In vitro grapevine plantlets grown under high salt concentrations showed a cultivar-dependent response for anthocyanin accumulation, which correlated with the expression of MYBA1-2, MYCA1 and WDR1 genes. These results suggest that MYCA1 may regulate ANR and UFGT and that this last control is easier to distinguish whenever MYBA genes are absent or in low abundance. Future studies should address the specific interactions of these proteins and their quantitative contribution to flavonoid synthesis in grape berries.

Job satisfaction among selected hospital CEOs.

The purpose of this study was to identify the contributors to job satisfaction of hospital chief executive officers (CEOs) using a multidimensional approach of demographic characteristics. environmental traits, and person environment fit traits. By analyzing the concept of hospital executive job satisfaction in a multidimensional approach, a more comprehensive model of the most salient determinants of job satisfaction was developed. CEOs ranked their performance highest on employee and staff relations and managerial team building and lowest on information management systems. The results of this study can be used to better understand the intricacies and uniqueness of being a hospital CEO as well as the professional and personal requirements of success.

Dose-response curve of human extensor digitorum brevis muscle function to intramuscularly injected botulinum toxin type A.

To develop a human model for measuring the effect of intramuscularly injected botulinum toxin, we injected both extensor digitorum brevis (EDB) muscles in 13 healthy volunteers with seven varying doses of botulinum toxin type A. We measured, pre- and postinjection, EDB M-wave amplitude, area, and mean rectified voltage (MRV) (obtained during maximal voluntary muscle activation). There was a logarithmic-appearing dose-response relationship between increasing doses of botulinum toxin and decline in EDB M-wave amplitude, area, and MRV. The decline was incrementally less at higher doses of toxin and appeared to level off at a maximal effect of 85 to 90% decrement from baseline (85 to 90% "paralysis") at 15 to 20 units. The peak toxin effect was present on day 6 postinjection. Measurement of EDB M-wave amplitude, area, and MRV is a simple objective method for quantifying the onset and degree of human muscle "paralysis" following botulinum-toxin injection.

A self-limited febrile illness produced in guinea pigs associated with oral administration of Legionella pneumophila.

The presumed route of human infection by Legionella pneumophila is inhalation. We investigated possible oral transmission of legionellosis in guinea pigs. Fifty-six guinea pigs (group 1) were given virulent L. pneumophila, serogroup 1, in drinking water. Fifty-nine guinea pigs (group 2) were inoculated with L. pneumophila via gastric intubation. Nineteen guinea pigs (group 3) were given heat-killed L. pneumophila in drinking water. Twenty-four guinea pigs (group 4, positive control) were inoculated intraperitoneally with L. pneumophila. Twenty-seven guinea pigs (group 5, negative control) were either intubated gastrically with phosphate-buffered saline or given drinking water without L. pneumophila. Sixty-six of 115 (57%) of the guinea pigs orally inoculated with viable L. pneumophila (groups 1 and 2) had a temperature greater than or equal to 103 degrees F and 8 of 115 (7%) had diarrhea, compared with 0 of 19 (0%) and 0 of 19 (0%), respectively, in group 3 and 1 of 27 (4%) and 0 of 27 (0%), respectively, in group 5. There were no fatalities in groups 1, 2, 3, and 5 compared with 15 of 24 (63%) in group 4. Groups 1, 2, and 4 consistently showed pneumonitis and splenitis. The pneumonitis of groups 1 and 2 was mild, predominantly interstitial, and mainly composed of macrophages; neither gross nor microscopic evidence of aspiration was seen. In group 1, 4 of 29 (14%) guinea pigs tested seroconverted to L. pneumophila compared with 0 of 7 (0%) in group 3 and 0 of 10 (0%) in group 5. In groups 1 and 2 combined, L. pneumophila was isolated from the lung of 5 of 57 (11%) guinea pigs and spleen of 5 of 47 (11%) guinea pigs compared with 0 of 14 guinea pigs in group 5. We conclude that viable L. pneumophila administered orally produces a self-limited febrile illness in guinea pigs.