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INTRODUCTION: Cannabis sativa L. is a well-recognized medicinal plant. Cannabis regulations in Argentina are insufficient to solve the problem of patient access to full-spectrum cannabis-based products. So, the market of artisanal products with unknown quality and dosage of cannabinoids is increasing, and so is the local demand and need for analyzing these products. However, much of the latest validated methodologies for cannabinoid quantification include expensive instrumentation that is not always available in laboratories of health institutions in Argentina. METHODS: The aim of this work was to develop and validate a simple and rapid HPLC-UV method for the identification and quantification of principal cannabinoids in cannabis resins, inflorescences, and medicinal oils using standard HPLC equipment. The cannabinoids selected for validation were cannabidiol acid (CBDA), cannabigerol (CBG), cannabidiol (CBD), cannabinol (CBN), delta-9-tetrahydrocannabinol (Δ9-THC), cannabichromene (CBC), and tetrahydrocannabinol acid (THCA). A method for the simultaneous identification and quantification of these 7 main cannabinoids was developed and then validated. Some data parameters were comparable to other reports with more sophisticated analytical instruments for the analysis of cannabis. The assessed limits of detection and the limits of quantitation ranged from 0.9 to 3.66 µg/mL and 2.78 to 11.09 µg/mL, respectively. The concentration-response relationship of the method indicated a linear relationship between the concentration and peak area with R2 values of > 0.99 for all 7 cannabinoids. RESULTS: The relative standard deviation (RSD%) varied from 2.34 to 4.82 for intraday repeatability and from 1.16 to 3.15 for interday repeatability. The percentage of recovery values was between 94 to 115% (resins) and 80 to 103% (inflorescence extract). The cannabis industry is growing rapidly, and there is a need for reliable testing methods to ensure the safety and efficacy of cannabis products. In addition, current methods for cannabinoid analysis are often time-consuming and expensive, while the HPLC-UV method herein reported is a simple, rapid, accurate, and cost-effective alternative for the analysis of cannabinoids in cannabis resins, inflorescences, and medicinal oils. CONCLUSION: This method will be proposed to be included in the Cannabis sativa L. monograph of the Argentine Pharmacopoeia.

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Larrea divaricata is a plant with antiproliferative principles. We have previously identified the flavonoid quercetin-3-methyl ether (Q-3-ME) in an ethyl acetate fraction (EA). Both the extract and Q-3-ME were found to be effective against the EL-4 T lymphoma cell line. However, the mechanism underlying the inhibition of tumor cell proliferation remains to be elucidated. In this work, we analyzed the role of nitric oxide (NO) in the induction of apoptosis mediated by Q-3-ME and EA. Both treatments were able to induce apoptosis in a concentration-dependent and time-dependent manner. The western blot analysis revealed a sequential activation of caspases-9 and 3, followed by poly-(ADP-ribose)-polymerase cleavage. EA and Q-3-ME lowered the mitochondrial membrane potential, showing the activation of the intrinsic pathway of apoptosis. Q-3-ME and EA increased NO production and inducible NO synthase expression in tumor cells. The involvement of NO in cell death was confirmed by the nitric oxide synthases inhibitor L-NAME. In addition, EA and Q-3-ME induced a cell cycle arrest in G0/G1 phase. These drugs did not affect normal cell viability. This data suggested that EA and Q-3-ME induce an increase in NO production that would lead to the cell cycle arrest and the activation of the intrinsic pathway of apoptosis. Copyright © 2016 John Wiley & Sons, Ltd.

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Vanillic acid is found at high concentrations in many plants used in traditional medicine. It has been associated with a variety of pharmacologic activities such as carcinogenesis inhibition, apoptosis and inflammation; however, it has become most popular for its pleasant creamy odor. Since there are few reports concerning the antinociceptive activity of this phenolic compound, the aim of this work was to study this activity in in vivo animal models. Vanillic acid was administered by the intraperitoneal route producing a dose-dependent inhibition of the acetic acid-induced writhing response (ED50: 9.3mg/kg). The antinociceptive activity was inhibited by the pretreatment with ondansetron and yohimbine, indicating that the serotoninergic and adrenergic systems could participate in the mechanism underlying the analgesic activity of vanillic acid. This compound was also demonstrated to interact with ASICs (Acid-sensing Ion Channels) as well as with TPRV1, TRPA1, and TRPM8 receptors in vivo. Furthermore, vanillic acid did not interfere with the locomotor function or motor coordination. The plasmatic phenolic content, analyzed by HPLC, showed that its t1/2 and AUC were 0.123h and 1.38µg·h/mL; respectively. In conclusion, vanillic acid might represent a potential therapeutic option for the treatment of pain.

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Larrea divaricata Cav. is a plant growing in South America. Both the infusion and a derived fraction (F1) of L. divaricata have proved to have immunomodulatory properties. Moreover, F1 can activate macrophages obtained from mice infected with Candida albicans. In this work, F1 was administrated to infected animals, and the state and type of activation of resident macrophages were studied. Results showed that F1 was able to activate macrophages obtained from infected mice by both classical and alternative pathways, probably by inducing a translocation of nuclear factor kappa-B. F1 increases not only the lysosomal activity of macrophages but also the production of phagosomal superoxide anion as a consequence of the activation of the Nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) complex. F1 induced an increase in the macrophage capacity to kill the fungus, which was reflected in a decrease in the levels of colonization of organs. A main flavonoid, kaempferol-3,4'-dimethylether, was identified in F1 by HPLC. This compound increased in vitro production of nitric oxide in heat-killed C. albicans-stimulated macrophages. The flavonoid could thus be considered one of the responsible molecules mediating the overall effects of F1 on the immune system in infected animals.

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Leukemia and lymphoma are a group of heterogeneous neoplastic disorder of white blood cells characterized by the uncontrolled proliferation and block in differentiation of hematopoietic cells. Nowadays, there is an interest in therapy with drugs of plant origin because conventional medicine can be inefficient or also results in side effects. Larrea divaricata Cav., is a plant widely distributed in Argentina that possess antiproliferative and antioxidant activities reported. Nordihydroguaiaretic acid (NDGA) was previously found in the plant and related to both antiproliferative and pro-proliferative actions on a lymphoma cell line. In order to demonstrate whether the presence of NDGA may be beneficial or not in the antiproliferative action of the aqueous extract, the extract of L. divaricata was submitted to a fractionation and fractions with and without NDGA were studied in a murine lymphocitic leukemia cell line (EL-4) proliferation. The effect of the most active fraction was studied in relation to H2O2 modulation and the synergistic action between compounds, found in fractions, was analyzed. The presence of NDGA was not a detonator for pro-proliferative action and its presence could be beneficial in low concentrations allowing a synergist antiproliferative action with other compounds.

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Larrea divaricata Cav. (jarilla) is a plant with well-documented applications in Argentinean folk medicine. In order to determine if the treatment with a purified fraction named F1 was capable to maintain a state of priming of macrophages after 15 days of mice infection with Candida albicans. Infected and uninfected mice were used. The effect of F1 on: cytosolic protein levels, apoptosis, phagocytosis, reactive oxygen species production, nitric oxide (NO), cell activity, lysosomal activity and the tissue fungal burden were studied. The results showed that F1 increased macrophages yeast phagocytosis and reactive oxygen species and NO production. All these effects were related to a decrease of cell activity and possible apoptosis. In conclusion, it was observed that F1 could induce a state of long-term activation of macrophages, since we observed increased activity of macrophages 15 days after infection, and it could be related to the elimination of C. albicans. These data may suggest that F1 fraction could be useful against disseminated candidiasis in patients and further studies on this field are desirable.

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Capsicum species, Solanaceae, have been used for centuries as food additives by populations of different regions of the world. Capsaicin (trans-methyl-N-vainillyl-nonenamide) is the main pungent compound extracted from the red hot pepper fruit (Capsicum spp.). The capsaicin content was determined by means of a HPLC method. The results showed that Capsicum chacoense Hunz., contains similar amounts of capsaicin (13.9 mg/100 g of dry fruit) in comparison with Capsicum baccatum L. (12.6 mg/100 g) and Capsicum annum L. (10.1 mg/100 g). Dichloromethane (CH2Cl2) and ethanol (EtOH) extracts of C. chacoense elicited a 46% and 38 % of inhibition on the arachidonic acid (AA) pathway in ear edema respectively while the CH2Cl2 and EtOH extracts of C. baccatum inhibited 52% and 35% the arachidonic acid response respectively.

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Larrea divaricata Cav. is widely distributed in Argentina. Aqueous extract, of its leaves, has documented antitumoral and immunomodulatory activities. In this study, the antioxidant activity of aqueous extract and a component, nordihydroguaiaretic acid was determined and compared using different assays. Both the aqueous extract and nordihydroguaiaretic acid exhibited antioxidant activity. However, results show that it is very likely that compounds other than nordihydroguaiaretic acid could be involved in the antioxidant activity of the extract. Since nordihydroguaiaretic acid is nephrotoxic and hepatotoxic agent, it is important to direct efforts toward the potential use of low-nordihydroguaiaretic acid L. divaricata extracts as antioxidant in foods.

Larrea divaricata Cav. está ampliamente distribuida en la Argentina. Se han documentado actividades antitumorales e inmunomoduladoras de los extractos acuosos de sus hojas. En este estudio, la actividad antioxidante del extracto acuoso y un componente, el ácido nordihidroguayarético, se determinaron y compararon mediante diferentes ensayos. Tanto el extracto acuoso como el ácido nordihidroguayarético mostraron actividad antioxidante. Sin embargo, los resultados muestran que es muy probable que otros compuestos diferentes al ácido nordihidroguayarético pudieran estar involucrados en la actividad antioxidante de los extractos. Dado que el ácido nordihidroguayarético es un agente nefrotóxico y hepatotóxico, es importante dirigir los esfuerzos hacia el uso potencial de extractos de L. divaricata con bajas cantidades de ácido nordihidroguayarético como antioxidantes en alimentos.

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Larrea divaricata is a plant widely used in folk medicine in Argentina. It has been demonstrated that an aqueous extract of L. divaricata possesses a biphasic effect on cell proliferation, at low concentrations exerts a stimulatory action and at high concentrations exerts anti-proliferative effects upon the T lymphoma BW 5147; therefore, we propose in this paper to test the effect of the extract 'in vitro' and 'in vivo' in another T-cell lymphoma named EL-4. It was analyzed 'in vitro' cell proliferation by tritiated thymidine uptake and the effect of the extract on tumors induced in mice analyzing tumor progression and survival.The results showed that the aqueous extract induced the proliferation of tumor cells at all the concentrations studied. The results 'in vivo' showed that the aqueous extract stimulated significantly the size of tumors and that untreated mice lived longer than those treated. It is important to be very careful when plant extracts are selected for the treatment of several diseases. Consequently, before using a plant extract, specific scientific studies must be undertaken on different models to certificate therapeutic and adverse effects. Moreover, it can be said that L. divaricata has a specific anti-tumor mechanism of action depending on the targets.

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OBJECTIVE: To compare the effects of intrathecal isobaric ropivacaine (IR) versus isobaric bupivacaine (IB) in a dose ratio of 3:2 in non-ambulatory urologic and orthopedic surgery. METHODS: One hundred and seventeen patients scheduled for surgery were randomized and assigned in a double-blind fashion to receive either 15 mg of IR (n = 58) or 10 mg of IB (n = 59) into the subarachnoid space. RESULTS: There were no differences about age, sex distribution, body mass index, type of surgical interventions and operative time. Both groups were similar respect to latency time and extension of sensory block. Complete motor blockade was reached in more than 90 % of patients of both groups ten minutes after the intrathecal injection and the length was significantly longer in the IB group (226.4 +/- 22.3 minutes versus 266.5 +/- 29.5, p < 0.001). There was no significant hypotension or bradicardia during the transoperative period. Three and a half hours after the end of the surgery, more patients of the IB group required intravenous analgesics and opiaceous derivates (p < 0.001). CONCLUSIONS: The motor blockade was longer in the IB group but postoperative analgesia was better in the IR group.

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