Herbal Blends Research Articles

Zyflamend, a unique herbal blend, induces cell death and inhibits adipogenesis through the coordinated regulation of PKA and JNK

ABSTRACT The prevalence of obesity and its comorbidities has sparked a worldwide concern to address rates of adipose tissue accrual. Recent studies have demonstrated a novel role of Zyflamend, a blend of natural herbal extracts, in regulating lipid metabolism in several cancer cell lines through the activation of the AMPK signalling pathway. Yet, the role of Zyflamend in adipogenic differentiation and lipid metabolism remains largely unexplored. The objective of this study is to investigate the effects of Zyflamend on white 3T3-MBX pre-adipocyte differentiation and elucidate the molecular mechanisms. We demonstrate that Zyflamend treatment altered cell cycle progression, attenuated proliferation, and increased cell death of 3T3-MBX pre-adipocytes. In addition, treatment with Zyflamend inhibited lipid accumulation during the differentiation of 3T3-MBX cells, consistent with decreased expression of lipogenic genes and increased lipolysis. Mechanistically, Zyflamend-induced alterations in adipogenesis were mediated, at least in part, through the activation of AMPK, PKA, and JNK. Inhibition of AMPK partially reversed Zyflamend-induced inhibition of differentiation, whereas the inhibition of either JNK or PKA fully restored adipocyte differentiation and decreased lipolysis. Taken together, the present study demonstrates that Zyflamend, as a novel anti-adipogenic bioactive mix, inhibits adipocyte differentiation through the activation of the PKA and JNK pathways. Abbreviation: 7-AAD: 7-amino-actinomycin D; ACC: acetyl-CoA carboxylase; AKT: protein kinase B; AMPK: AMP-activated protein kinase; ATGL: adipose triglyceride lipase; C/EBPα: CCAAT-enhancer binding protein alpha; DMEM: Dulbecco’s Modified Eagle Medium; DMSO: dimethyl sulphoxide; DTT: dithiothreitol; EGTA: ethylene glycol-bis-(2-aminoethyl)-N,N,N’,N’-tetraacetic acid; ERK: extracellular signal–regulated kinases; FASN: fatty acid synthase; FBS: foetal bovine serum; GLUT: glucose transporter; HSL: hormone-sensitive lipase; IR: insulin receptor; IRS: insulin receptor substrate; JNK: c-JUN N-terminal kinase; MGL: monoacylglycerol lipase; NaF: sodium fluoride; NF-κB: nuclear factor kappa-light-chain-enhancer of activated B cells; PBS: phosphate buffered- saline; PCB: pyruvate carboxylase; PDE: phosphodiesterase; PKA: protein kinase cAMP-dependent; PMSF: phenylmethylsulfonyl fluoride; PPARγ: perilipin peroxisome proliferator-activated receptor gamma; PREF-1: pre-adipocyte factor 1; PVDF: polyvinylidene fluoride; RIPA: radio-immunoprecipitation assay; SDS-PAGE: sodium dodecyl sulphate polyacrylamide gel electrophoresis; SEM: standard error of the mean; SOX9: suppressor of cytokine signalling 9; TGs: triacylglycerols.

Four cases of death involving the novel synthetic cannabinoid 5F-Cumyl-PEGACLONE

PurposeCumyl-PEGACLONE was the first synthetic cannabinoid (SC) with a γ-carbolinone core structure detected in forensic casework and, since then, it has dominated the German SC-market. Here the first four cases of death involving its fluorinated analog, 5F-Cumyl-PEGACLONE, a recently emerged γ-carbolinone derived SC, are reported.MethodsComplete postmortem examinations were performed. Postmortem samples were screened by immunoassay, gas chromatography mass spectrometry (GC–MS) or liquid chromatography tandem mass spectrometry. For quantification of SCs, the standard addition method was employed. Herbal blends were analyzed by GC–MS. In each case of death, the Toxicological Significance Score (TSS) was assigned to the compound.Results5F-Cumyl-PEGACLONE was identified at concentrations ranging 0.09–0.45 ng/mL in postmortem femoral blood. In case 1, signs of hypothermia and kidney bleedings were noted. Despite a possible tolerance due to long term SC use, a TSS of 3 was assigned. In case 2, an acute heroin intoxication occurred and a contributory role (TSS = 1) of 5F-Cumyl-PEGACLONE was suggested. In case 3, a prisoner was found dead. GC–MS analysis of herbal blends, retrieved in his cell together with paraphernalia, confirmed the presence of 5F-Cumyl-PEGACLONE and a causative role was deemed probable (TSS = 2). In case 4, the aspiration of gastric content due to a SC-induced coma was observed (TSS = 3).Conclusions5F-Cumyl-PEGACLONE is an emerging and extremely potent SC which raises serious public health concerns. A comprehensive analysis of circumstantial, clinical, and postmortem findings, as well as an in-depth toxicological analysis is necessary for a valid interpretation and for the assessment of the toxicological significance.

Study the effect of herbal mixture plants extract on blood sugar level in normal and experimentally diabetic mice

The use of medicinal plants for the management of diabetes mellitus is an old practice that has become even more relevant in a modern perspective. The present work was designed to evaluate the effect of a mixture of three medicinal plants which used in folk medicine in Iraq, These plants are (Aloe vera, Artemisia herba alba, and Teucrium polium) on the levels of blood glucose in normal and alloxan-induced diabetic mice, The aqueous extract of the herbal mixture was prepared and chemical detection of phenols, flavonoids, tannins, terpenes, steroids, glycosides, and saponins was carried out. Results revealed that the aqueous extract contains phenols, flavonoids, tannins, terpenes, glycosides, and saponins compounds. Evaluations of the parameters of our study were carried out on both standard and alloxan-induced diabetic mice. Thirty male mice were randomly divided into six equal groups: Group I (control): was kept as control negative mice treated with only distilled water. Group II: normal mice treated with aqueous extract of an herbal mixture at a dose (500 mg/kg/day).Group III: normal mice treated with aqueous extract of an herbal blend at a dose (250 mg/kg/day). The other 3 groups were subcutaneously administered a single dose (100 mg/kg) of alloxan to induce experimental diabetes. Groups IV (Diabetic): was kept as control positive, alloxan-induced diabetic mice treated with only distilled water. Groups V: alloxan-induced diabetic mice treated with aqueous extract of an herbal mixture at a dose (500 mg/kg/day). Group VI: alloxan-induced diabetic mice treated with aqueous extract of an herbal blend at a dose (250 mg/kg/day), respectively, for ten days. Results showed that normal mice treated with aqueous extract has no significant change in body weights and blood glucose level except those treated with a high dose of aqueous extract since they exhibited a significant decrease (P ≤ 0.05) in blood glucose level. The results indicated a significant reduction in glucose level in diabetic mice after treatment with a high dose of aqueous extract of the herbal mixture. In conclusion, our results support that the aqueous extract of these plant exhibits anti-diabetic as compared with each plant alone, where we tested each of these plants in previous studies.

Pharmacological and Behavioral Effects of the Synthetic Cannabinoid AKB48 in Rats.

AKB48 is a designer drug belonging to the indazole synthetic cannabinoids class, illegally sold as herbal blend, incense, or research chemicals for their psychoactive cannabis-like effects. In the present study, we investigated the in vivo pharmacological and behavioral effects of AKB48 in male rats and measured the pharmacodynamic effects of AKB48 and simultaneously determined its plasma pharmacokinetic. AKB48 at low doses preferentially stimulated dopamine release in the nucleus accumbens shell (0.25 mg/kg) and impaired visual sensorimotor responses (0.3 mg/kg) without affecting acoustic and tactile reflexes, which are reduced only to the highest dose tested (3 mg/kg). Increasing doses (0.5 mg/kg) of AKB48 impaired place preference and induced hypolocomotion in rats. At the highest dose (3 mg/kg), AKB48 induced hypothermia, analgesia, and catalepsy; inhibited the startle/pre-pulse inhibition test; and caused cardiorespiratory changes characterized by bradycardia and mild bradipnea and SpO2 reduction. All behavioral and neurochemical effects were fully prevented by the selective CB1 receptor antagonist/inverse agonist AM251. AKB48 plasma concentrations rose linearly with increasing dose and were correlated with changes in the somatosensory, hypothermic, analgesic, and cataleptic responses in rats. For the first time, this study shows the pharmacological and behavioral effects of AKB48 in rats, correlating them to the plasma levels of the synthetic cannabinoid.Chemical Compound Studied in This Article: AKB48 (PubChem CID: 57404063); AM251 (PubChem CID: 2125).

Novel halogenated synthetic cannabinoids impair sensorimotor functions in mice

JWH-018-Cl, JWH-018-Br and AM-2201 (JWH-018 halogenated-derivatives; JWH-018-R compounds) are synthetic cannabinoid agonists illegally marketed as “Spice”, “K2”, “herbal blend” and research chemicals for their cannabis-like psychoactive effects. In rodents, JWH-018 and its halogenated derivatives reproduce the typical effects of Δ9-tetrahydrocannabinol (Δ9-THC), i.e. hypothermia, analgesia, hypolocomotion and akinesia. Yet, the effects of JWH-018-R compounds on sensorimotor functions are still unknown. This study was designed to investigate the effect of an acute intraperitoneal (i.p.) administration of JWH-018-R compounds (0.01–6 mg/kg) on sensorimotor functions in mice and to compare them to those caused by the reference compound JWH-018 and Δ9-THC. A well validated battery of behavioral tests was used to investigate the effects of these synthetic cannabinoids on the visual, auditory and tactile responses in mice, while the pre-pulse inhibition (PPI) test was used to investigate their effect on sensorimotor gating. The effect of the synthetic cannabinoids on spontaneous locomotion was also measured by a video tracking analysis to assess potential cannabinoid-induced motor impairment. Results showed that, similarly to JWH-018, systemic administration of JWH-018-R compounds inhibits sensorimotor and PPI responses at lower doses (0.01-0.1 mg/kg) and reduced spontaneous locomotion at intermediate/high doses (1–6 mg/kg). All effects were prevented by the administration of the selective cannabinoid CB1 receptor antagonist/inverse agonist AM-251 thus confirming a CB1 receptor-mediated action. Finding that lower doses of JWH-018-R compounds selectively impair sensorimotor and PPI responses without affecting locomotion should be carefully considered to better understand the potential danger that halogenated-derivatives of JWH-018 may pose to public health, with particular reference to decreased performance in driving and hazardous works.

Ocena działania przeciwcukrzycowego ekstraktów otrzymanych z wybranych surowców roślinnych

Introduction. Diabetes is characterized by chronic hyperglycaemia, resulting from a defect in insulin secretion or abnormal functioning of this hormone. The consequence of the lack of insulin is the disturbed metabolism of carbohydrates, proteins and fats. Chronic hyperglycemia causes damage and dysfunction of many organs. One of the ways to support the treatment of diabetes is the use of plant preparations. Aim. The aim of the study was to examine and compare the biological (α-glucosidase and antioxidative) activity and the content of polyphenols and flavonoids in selected plants: Anserinae herba, Alchemillae herba and Fragariae folium, included in herbal tea blends used in diabetes. Material and methods. The content of polyphenols was determined by applying the Folin-Ciocalteu reagent and the flavonoid content by the AlCl3 method. The antioxidant properties were tested using the reagent method DPPH and CUPRAC, while the inhibitory activity of α-glucosidase by measuring the absorbance of p-nitrophenol from PNPG released by hydrolysis of sugars by the enzyme. Results. High total polyphenols content in the tested plant extracts was found, amounting to 121.30; 98.30; 71.66 mg gallic acid equivalent/g for Alchemillae herba, Fragariae folium and Anserinae herba, respectively, and high flavonoid content (17.56, 12.25, 15.48 mg quercetin equivalent/g, respectively). Antioxidant activity for extracts with a concentration of 0.312 mg/ml was from 46.22 to 79.71% (in the analysis with DPPH radical), while the absorbance value of these tests determined by the CUPRAC method was A = 0.49-1.01. The effect was weaker than vitamin C. The results indicated a strong inhibitory ability of α-glucosidase, which was > 98% for all tested extracts at the concentration of 0.1 mg/ml, and was many times higher than the standard acarbose analyzed at the above concentration, and comparable with the acrabose activity at a concentration of 10 mg/ml. Conclusions. The inhibitory α-glucosidase and antioxidant activity indicates the existence of this mechanism included in analyzed plant extracts, as ingredients herbal tea blends of used in diabetes.

Histochemical assays of secretory trichomes and the structure and content of mineral nutrients in Rubus idaeus L. leaves

Leaves of Rubus idaeus are a raw material, ingredients of herbal blend, and a source of antioxidants. There are no data concerning histochemistry of trichomes, and little is known about the leaf structure of this species. The aim of this study was to determine the histochemistry of active compounds and the structure of glandular trichomes, micromorphology, anatomy, and ultrastructure of leaves as well as content of elements. To determine the histochemistry of glandular trichomes, different chemical compounds were used. The leaf structure was analysed using light, scanning, and transmission electron microscopes. The content of elements was determined with atomic absorption spectrometry, and the microanalysis of the epidermis ultrastructure was carried out with a transmission electron microscope equipped with a digital X-ray analyser. In glandular trichomes, polyphenols, terpenes, lipids, proteins, and carbohydrates were identified. The main elements in the ultrastructure of the epidermis were Na, Mo, Se, Ca, and Mg. In dry matter of leaves, K, Mg, Ca, P, and Fe were dominant. Infusions from leaves are safe for health in terms of the Cd and Pb concentrations. Leaves can be a valuable raw material. Non-glandular trichomes prevent clumping of mixed raw materials in herbal mixtures.

Detection of the recently emerged synthetic cannabinoid 4F-MDMB-BINACA in "legal high" products and human urine specimens.

Synthetic cannabinoids (SCs) remain one of the largest groups of new psychoactive substances (NPS) on the European drug market. Although the number of new derivatives occurring on the market has dropped in the last two years, newly emerging NPS still represent a challenge for laboratories performing forensic drug analysis in biological matrices. The newly emerged SC 4F-MDMB-BINACA has been reported by several law enforcement agencies in Europe and the USA since November 2018. This work aimed at revealing urinary markers to prove uptake of 4F-MDMB-BINACA and differentiate from the use of structurally similar SCs. Phase-I metabolites detected in human urine specimens were confirmed by phase-I metabolites generated in vitro using a pooled human liver microsomes (pHLM) assay. Seized materials and test-purchased "legal high" products were analyzed by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-quadrupole-time-of-flight-mass spectrometry (LC-qToF-MS). Human urine specimens and pHLM assay extracts were measured with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) and confirmed by LC-qToF-MS. In January 2019, the Institute of Legal Medicine in Erlangen (Germany) identified 4F-MDMB-BINACA in three herbal blends. During the same time period, the described SC was identified in a research chemical purchased online. Investigation of phase-I metabolism led to the metabolites M10 (ester hydrolysis) and M11 (ester hydrolysis and dehydrogenation) as reliable urinary markers. Widespread distribution on the German drug market was proven by analysis of urine samples from abstinence control programs and by frequent detection of 4F-MDMB-BINACA in "herbal blends" and "'research chemicals" purchased via the Internet.

A Novel Herbal Composition LI12542F6 Improves Muscle Endurance and Strength in Resistance Trained Males (P23-003-19)

ObjectivesThis study was designed to evaluate the efficacy of an herbal blend composed of Sphaeranthus indicus and Mangifera indica extracts to improve muscle health. MethodsIn a randomized, double-blind, placebo-controlled study, 40 male participants of 18–40 yrs age, were assigned to receive 650 mg of LI12542F6 (A) or Placebo (P) for 56 days. The primary endpoint was change from baseline in muscle strength, assessed by 1-RM Bench and Leg presses. Secondary endpoints included muscle endurance, time to exhaustion, muscle size, body composition and free testosterone. Safety was also assessed. ResultsSignificant (P < 0.0001) increases in change from baseline for strength (bench and leg presses and grip strength) were observed in the A vs. P group beginning at 14 d and continuing throughout the study. Muscle endurance and time to exhaustion were similarly increased by 14 d and continued for the remainder of the study. By day 56, the mid upper arm circumference was statistically increased in A vs. P (P < 0.05) in left and right arms. ConclusionsSignificant improvements in muscle endurance and strength were observed in resistance trained healthy men consuming LI12542F6 daily for 56 days. Further research will elucidate efficacy in other populations. Funding SourcesLaila Nutraceuticals.

A Randomized, Double-Blind, Placebo-Controlled Study of a Blend of Herbal Extracts Taken Once Per Day for Weight Loss in Healthy Volunteers

Aims: We previously demonstrated that a blend of herbal extracts (Weighlevel®; a mixture of extracts from the leaves of Alchemilla vulgaris, Olea europaea, Mentha longifolia and from the seeds of Cuminum cyminum) taken 3 times per day produces weight loss in preclinical and clinical studies. The aim of the present study was to test the efficacy of a new slow-release formulation (Weighlevel® One) taken once per day on change in body weight and related measures.&#x0D; Study Design: Randomized, double-blind, placebo-controlled study.&#x0D; Place and Duration of Study: Health Clinics in Copenhagen, Denmark between 7 January 2016 and 5 March 2016.&#x0D; Methodology: Thirty-six adult subjects were randomized to consume the herbal blend (n = 20) or placebo (n = 16) once per day for 8 weeks. Weight and waist circumference were assessed weekly. The primary endpoint was the change from baseline in body weight for the herbal blend group compared with placebo. Secondary assessments included waist circumference, appetite, craving, bowel health, and safety and tolerability.&#x0D; Results: After 8 weeks, the herbal blend group lost an average of 3.7 kg (95% CI of 3.0 to 4.5 kg); whereas the placebo group lost 0.1 kg (95% CI of -0.7 to 1.0 kg). This difference in mean weight loss between the herbal blend and placebo groups was statistically significant (P &lt;.001). A statistically significant reduction in waist circumference was also observed for the herbal blend compared with placebo (P &lt;.001). The herbal blend was well tolerated; no adverse events were reported.&#x0D; Conclusion: Daily administration of this blend of herbal extracts, administered once daily, may produce weight loss.

Cytotoxicity of the synthetic cannabinoids 5C-AKB48, 5F-MDMB-PINACA, ADB-CHMINACA, MDMB-CHMICA and NM-2201 in A549 and TR146 cell lines

The aim of the present study was to evaluate the cytotoxicity of five synthetic cannabinoids in two cell lines, for which both reference standards and herbal blends were available. An in-house smoking device was developed to produce smoke condensates. The cell viability of reference standards, herbal blend extracts and smoke condensates was measured using an MTT assay in the A549 lung carcinoma cell line and the TR146 buccal carcinoma cell line. Damiana extracts and damiana smoke condensates were also tested for cytotoxicity. For the reference standards of 5F-MDMB-PINACA, ADB-CHMINACA and MDMB-CHMICA, a significant concentration-dependent decrease in cell viability was observed. ADB-CHMINACA and MDMB-CHMICA extracts were more potent than the damiana extract, whereas the NM-2201 blend extract abrogated the effects of the damiana leaves. Compared with damiana smoke condensate, MDMB-CHMICA smoke condensate had higher sensitivity, while ADB-CHMINACA smoke condensate was less potent. This is the first study to investigate the comparative cytotoxicity of reference standards, herbal blend extracts and smoke condensates of synthetic cannabinoids. The data showed that the presence of damiana plant contributed to enhancing the cytotoxicity of 5C-AKB48, 5F-MDMB-PINACA, and MDMB-CHMICA. The plant effects were generally more marked for smoke condensates. MDMB-CHMICA was the most potent substance. The NM-2201 reference standard and extract led to an increase in cell viability as compared to the control. This is of special interest, because the NM-2201 herbal blend extract seemed to abrogate the effects of the damiana leaf extracts. Further studies would be necessary to assess potential procarcinogenic or tumorigenic effects of NM-2201.

Mycotoxins in herbal teas marketed in Latvia and dietary exposure assessment

ABSTRACTThe occurrence of 12 mycotoxins has been analysed by liquid chromatography – time of flight mass spectrometry in the batch of 60 herbal teas purchased from drugstores in Latvia. Among the dry tea samples, 90% were positive for one to eight mycotoxins. Enniatin B and deoxynivalenol (DON) were the most frequently detected mycotoxins in 55% and 45% of the samples, respectively. DON reached the highest level, from 129 µg kg−1 in herbal blend to 5,463 µg kg−1 in wormwood tea. Ochratoxin A (OTA) and aflatoxin B1 (AFB1) were found in 10% and 20% of the samples at the concentrations ranged between 2.99–30.3 µg kg−1 and 3.40–23.7 µg kg−1. Studies of the tea infusion process indicated that 32–100% of DON and zearalenone present in dry teas were extracted into the infusions. Dietary exposure assessment was performed, using the determined mycotoxin levels and the available consumption data.

Sacharomyces cerevisiae as a screening tool to assess cathinones’ toxicity

Introduction: Novel psychoactive substances (NPS) are sold on Internet under the appearance of herbal blends or bath salts as an alternative to cannabis, khat and ephedra. Synthetic cathinones are artificial mimetics of khat and like methamphetamine are indirect agonists of dopamine, serotonin and noradrenaline receptors [1]. Ninety-seven fatal cases of intoxication by ingestion of mephedrone (4-methylmethcathinone), one of the types of synthetic cathinone derivatives, were reported on literature since 2010 [2]. New synthetic cathinones emerges in the street market and internet on a regular base and the properties of this substances change regularly, due to modification on the molecules, to avoid legislation. This makes almost impossible to characterize synthetic cathinones toxicological profiles on a short time scale, mostly due to the typical time-consuming experimental assays. In here we propose the use of Saccharomyces cerevisiae, which has been proven an invaluable model to study the fundamental molecular mechanisms involved in several human diseases [3], growth curves in the presence of synthetic cathinones to get an indicative measure of its toxicity. Materials and methods: BY4741 yeast cells were diluted to a standardized OD600 nm =2×10−3 (∼1.67×10-5 cells/mL) in YNB medium (Yeast Nitrogen Base w/o Amino Acids 0.67%, glucose 2%, required amino acids) and grown at 30 °C. Buphedrone was acquired from the best source available. The cytotoxic potential of synthetic cathinones was assessed by monitoring the growth of Saccharomyces cerevisiae BY4741 in the presence and absence of substances. Yeast growth curves (O.D.600nm versus growth time) were compared. Results: Figure 1 shows that yeast growth in the presence of 75 mM cat is lower when compared to the growth curve in the absence of the substance. These preliminary results points that cathinones inhibits yeast growth. Figure 1. Growth curves results in Saccharomyces cerevisiae BY4741 in the presence of 75 mM of buphedrone. Discussion and conclusions: Saccharomyces cerevisiae has been used previously to test the toxicity of pharmacological drugs. Our results show that yeast is sensitive to cathinones suggesting that this system has potential to assess the possible toxicity of these legal highs. This rapid and inexpensive methodology may allow high throughput screening of cathinones’ toxicity, which might help the legislators to schedule NPS avoiding expected public health problems.

Should we legalize marijuana? Ten years of learning from JWH-018 first seizure

Introduction: The medical use of cannabis predates history, finding its first historical root in ancient China 4700 years ago. The main psychoactive component of cannabis is Δ9-tetrahydrocannabinol (Δ9 -THC) first isolated by Gaoni and Mechoulam [1]. In the 1960’s Richard Nixon started a war on drugs both to control the internal US movements against Vietnam war and for racial reasons. In fact, citing JD Ehrlichman, Nixon assistant for domestic affairs, “getting the public to associate the hippies with marijuana and blacks with heroin and then criminalizing both heavily, we could disrupt those communities” [2]. Due to cannabis control by UN acts of 1961 and 1971, in the end of 70’s Pfizer© and Lilly© start developing synthetic cannabinoids (SC) for clinical purposes [3]. SC have similar effects of Δ9 -THC and cannabidiol, (CBD) presented in marijuana. JWH-018 a SC was synthetized by J.W.Huffman in 1995 as a drug to treat cancer [4]. This molecule was the active substances of herbal blends sold on smartshops, according to a report from the pharmaceutical company THC Pharm in 2008 [5]. After this almost 200 new SC have been reported by the early warning system of the EMCDDA [6]. The ugly true is that, in early years of XXI century, products containing SC have been sold in Europe and marketed as ‘legal’ replacements for cannabis. Moreover a 2009 survey conducted among 1463 students aged between 15 and 18 in Frankfurt found that around 6% of the students reported having used ‘Spice’ at least once [5]. New psychoactive substances (NPS) in general and SC in particular may pose health and social risks to our society that demands an open and scientific discussion about marijuana legalization. Materials and methods: A review was carried out on PubMed, EMCDDA and United Nations sites to gather scientific data about cannabis and SC effects on human health. Results: A study, from 1843, suggested that cannabis sativa “possess very striking powers as an antispasmodic, as a nervine stimulant, and as an anodyne and hypnotic, and in some respects to excel opium in virtue” [7]. 175 years after this report, studies on neurological uses of Δ9 –THC and CBD reports positive outcomes on malignant brain tumors, Parkinson’s, Alzheimer’s, multiple sclerosis and neuropathic pain. In addition, psychiatric and mood disorders, such as schizophrenia, anxiety, depression, and posttraumatic stress also seem to benefit from the use of phytocannabinoids [8]. A great number of papers and clinical reports show that Δ9 –THC and CBD is beneficial clinically. Actually, Dronabinol and Nabilone are two cannabinoids synthetized and commercialized by pharmaceutic industry with clinical usage. Discussion and conclusions: SC molecules appears in the street and internet market on a regular base. The new emerging molecules are slightly different from the previous to avoid legislation. This makes almost impossible to characterize new SC toxicological profiles on a sensible time scale. The control of marijuana have been having consequences on consume of SC with reported fatalities [9] which are absent in marijuana consume. Besides, control of cannabis sativa has strong consequences in scientific research. It is time to face scientifically the pros and cons of marijuana legalization for a conscious political decision.

Advantageous Extraction, Cleanup, and UHPLC-MS/MS Detection of Patulin Mycotoxin in Dietary Supplements and Herbal Blends Containing Hawberry from Crataegus spp.

Patulin (PAT) is a highly genotoxic mycotoxin still found as the common contaminant of various kinds of spoiled fruits and related commodities which are often endorsed as the health-enhancing products. Thus, a fast and convenient liquid-solid extraction followed by a solid-phase cleanup with the MycoSep®228 AflaPat multifunctional column was used for the highly efficient isolation of PAT with an average recovery of 112.7% from commercial dietary supplements and herbal blends formulated with dried hawberry. Analysis of the PAT content was carried out using gradient elution with a Synergi Polar C18 column (150 × 2 mm, 4 μm) and UHPLC system equipped with a mass spectrometer. PAT was detected in all (n=14) commercial single-component dietary supplements formulated with dried hawberry belonging to Crataegus monogyna and/or Crataegus laevigata. Similarly, PAT was detected in 67% of the studied multicomponent commercial herbal blends (n=6) that contained—in addition to hawberry—different amounts of apple, chokeberry, elderberry, hibiscus, or mallow. Moreover, the PAT content was determined in the hawberry collected from the mature wild hawthorn trees belonging to three botanical species, Crataegus monogyna Jacq., Crataegus laevigata (Poiret) DC, and Crataegus rhipidophylla Gand, growing in the recreational forest areas and in the law-protected state national forest park in Poland. In conclusion, to prevent PAT accumulation and reduce the health risk of consumers in globalizing markets, the implementation of improved cultivation/processing practices of hawthorn trees and hawberry as well as increased analytical control related to the presence of PAT in dietary supplements and herbal blends formulated with fresh, dried, or frozen hawberry should be urgently recommended.

DEVELOPMENT AND CHARACTERIZATION OF BIOCOMPATIBLE POLYHYDROXY BUTYRATE IMPREGNATED WITH HERBAL PLANTS AGAINST WOUND HEALING ACTIVITY ON IN VIVO ANIMAL MODEL

Objective: The current study is to evaluate the antimicrobial, antioxidant, anti-inflammatory, and in vitro cytotoxicity activities of polyhydroxybutyrate (PHB) and to develop the herbal impregnated PHB cast film for wound healing activities using Albino Wistar rat model.&#x0D; Methods: PHB produced by Azotobacter chroococcum A3 strain was synthesized and characterized (previous study). The PHB was subjected to various biocompatibility studies such as antimicrobial, antioxidant, and anti-inflammatory studies. The PHB was also subjected to cytotoxicity study by (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. PHB films were made using different combinations of plant and algal blends (herbal blends). The herbal blends of PHB films were evaluated for in vivo wound healing activity using Albino Wistar rats.&#x0D; Results: The turmeric impregnated PHB showed the highest result for antimicrobial with 27.25±0.23 mm against skin pathogens and antioxidant activity with the highest percentage of inhibition of 76%. The result predicts that PHB will not let to any toxic substances rather it acts as a chemoprotective agent followed by the inhibitory concentration value was found to be 1.56 μg/ml for 100 μg. The in vivo study showed better wound healing activity for PHB blended with 2% turmeric leaf and rhizome cast film. Whereas the wound healing activity of control and crude PHB was 90.4±0.4 and 91.3±0.56 respectively.&#x0D; Conclusion: The results from the present study showed that PHB can act as a good candidate for drug carrier and it is biocompatible in living cells.

5F-Cumyl-PINACA in \u2018e-liquids\u2019 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

PurposeIn 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.MethodsThe 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.ResultsMetabolism 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.ConclusionsThe 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.

Efficacy of a novel herbal formulation for weight loss demonstrated in a 16-week randomized, double-blind, placebo-controlled clinical trial with healthy overweight adults.

To re-evaluate the weight loss efficacy of LI85008F in healthy overweight adults via a 16-week randomized, double-blind, placebo-controlled clinical study. One hundred and forty overweight participants (body mass index [BMI] 27-29.9 kg/m2 , 29.3% male; ages 21-50 years) were randomized into placebo (n =70) and LI85008F (n =70) groups. The participants received either 900 mg/d of LI85008F in two divided doses or two identical placebo capsules. In addition, participants were counselled to follow an ~1800 kcal/d diet and to engage in walking for 30 min, 5 d/wk throughout the study. At the end of the trial period, the LI85008F supplemented group showed significant reductions in body weight (5.36 ± 1.769 vs. 0.87 ± 1.381 kg; P < 0.0001) and BMI (2.05 ± 0.693 vs. 0.34 ± 0.559 kg/m2 ; P < 0.0001), compared with placebo. Significant reductions in waist and hip circumferences, and a 2.08-fold reduction of waist/hip ratio, were noted in the LI85008F supplemented group. LI85008F supplementation also resulted in significant improvements in lipid profiles, compared with the placebo; low-density lipoprotein (LDL) cholesterol decreased, while high-density lipoprotein (HDL) cholesterol increased, resulting in a significantly improved LDL/HDL ratio. No major adverse events were reported by the participants during the study. The unique herbal extract blend LI85008F, combined with modest calorie restriction and physical activity, is well tolerated, safe, and effective for weight management in overweight men and women.

Profiling of new psychoactive substances by using stable isotope ratio mass spectrometry: Study of the synthetic cannabinoid 5F-PB-22.

In this paper, the results of a pilot study on the profiling of the synthetic cannabinoid receptor agonist 5F-PB-22 (5F-QUPIC, pentylfluoro-1H-indole-3-carboxylic acid-8-quinolinyl ester) via isotope ratio mass spectrometry are presented. It focuses on δ13 C, δ15 N and δ2 H isotope ratios, which are determined using elemental analyser (EA) and high temperature elemental analyser (TC/EA) coupled to an isotope ratio mass spectrometer (IRMS). By means of a sample of pure material of 5F-PB-22, it is shown that the extraction of 5F-PB-22 from herbal material, a rapid clean-up procedure, or preparative column chromatography had no influence on the isotope ratios. Furthermore, 5F-PB-22 was extracted from 14 different herbal blend samples ("Spice products" from police seizures) and analysed via IRMS, yielding 3 clusters containing 7, 5, and 2 samples, distinguishable through their isotopic composition, respectively. It is assumed that herbal blends in each cluster have been manufactured from individual batches of 5F-PB-22.

The influence of selected feed additives on mineral utilisation and bone characteristics in laying hens

Abstract The trial with 240 caged ISA Brown laying hens was performed to evaluate the effect of selected feed additives on mineral utilisation as well as biomechanical (breaking strength, yielding load, stiffness) and geometrical (cortex thickness, cross-section area, weight, length) indices of tibia and femur bones. At 26 wks of age the layers were randomly assigned to 10 treatments with 12 replicates (cages) of two birds. In the study a 2 × 5 experimental scheme was used i.e. to 70 wks of age, the layers were fed isocaloric and isonitrogenous experimental diets containing reduced (3.20%) or standard (3.70%) Ca level. The diets with both Ca levels were either not supplemented, or supplemented with the studied feed additives i.e. sodium butyrate, probiotic bacteria, herbal extract blend and chitosan. There were no statistically significant effects of the experimental factors on the indices of the tibia bones. However, the diet with reduced Ca level decreased bone breaking strength, yielding load, stiffness, and mineralisation of the femur bones (P&lt;0.05). The majority of used feed supplements, i.e. probiotic, herb extracts, and chitosan, increased biomechanical indices (breaking strength and yielding load) and mineralisation of the femur bones (P&lt;0.05). Neither dietary Ca level nor feed additives affected dry matter, organic matter, ether extract, N-free extracts, crude fibre and ash digestibility, and P retention and excretion; however, Ca excretion and retention was lower in the hens fed the diets with reduced Ca level (P&lt;0.05). Relative Ca retention (Ca retained as % of Ca intake) was improved by diet supplementation with probiotic, herb extracts and chitosan (P&lt;0.05). In conclusion, this study has shown that decreased Ca dietary level (3.20%) can negatively affect bone quality in layers, while probiotic, herb extracts and chitosan addition may improve the selected biomechanical indices of the femurs, irrespective of Ca dietary concentration.