Toxicological safety of VOHO Hemp Oil; a supercritical fluid extract from the aerial parts of hemp

VOHO Hemp Oil (Verdant Nature LLC (in collaboration with HempFusion)) is an extract of the aerial parts of hemp (Cannabis sativa L) manufactured using a supercritical CO2 extraction process. The results of four safety studies are reported here including a bacterial reverse mutation assay, an in vivo mammalian micronucleus study, a maximum tolerated dose study in rats and a 90-day repeat dose subchronic toxicity study in rats. VOHO Hemp oil can contain up to 30% phytocannabinoids and less than 0.2% is tetrahydrocannabinol (THC). VOHO Hemp Oil was found to be non-mutagenic in the bacterial reverse mutation assay and was negative for inducing micronuclei in the rat bone marrow micronucleus assay. The maximum tolerated dose in male and female Wistar rats was 2250 mg/kg bw/day. A 90-day repeat dose study was conducted in male and female Wistar rats according to OECD Guideline 408 and included a 21-day recovery period. The doses used in the study were 0, 25, 90 and 324 mg/kg bw per day in the main study, and in the recovery phase a control and 324 mg/kg bw/day group were included. One mortality was reported during the study, a high dose female, and test substance-related adverse clinical signs were reported in the high dose group. Other test substance-related changes noted in the high dose group included changes in body weights, activated partial thromboplastin time (APTT) values, and in absolute and relative organ weights. Based on the results of the study, the no observed adverse effect level (NOAEL) for VOHO Hemp Oil was determined to be 90 mg/kg bw/day in both male and female Wistar rats.

Passive Measurements of the Refractive Index of Liquid Nitrogen and Free-flowing substances by the Prism Method in the Millimeter Wavelength Range

It is proposed to use the classical prism method in the millimeter wavelength range for measuring the refractive index of liquid and free-flowing substances, including mixtures with particle sizes comparable to the wavelength. The method is implemented using a hollow radio transparent rectangular prism filled with a test substance. The measurements were carried out in the thermal radiation mode using radiometers with horn-lens antennas at frequencies of 37.5 and 94 GHz. To measure the deflection of the refracted beam, a linear scanner with a black body mounted on it, cooled with liquid nitrogen, was used. The distance between the prism and the scanner was 1 m. The refractive index of liquid nitrogen, sand, gravel, marble chips and granular polyethylene were measured. Using the refractive formula and the Landau-Lifshitz-Looeng formula for calculating the dependence of the refractive index of binary mixtures on the bulk density of particles, estimates of the refractive index of the material of the particles that make up the substances under study are obtained. They are in satisfactory agreement with the known experimental data for quartz, feldspar, granite, and marble.

Pits of Date Palm: Bioactive Composition, Antibacterial Activity and Antimutagenicity Potentials

Palm date pits as a source of bioactive chemicals have received only a limited amount of attention. This study was conducted to determine the bioactive components of Ajwa date pits, as well as their antibacterial and antimutagenic activities. Therefore, chromatographic techniques were used to extract and isolate the phenolic compounds in date pits, which were further elucidated by using spectroscopic techniques (FTIR and NMR) for identification of most abundant bioactive metabolites. Data showed that the methanolic extract of date pits contained high levels of total phenols (17.38 mg GA/g) and flavonoids (5.324 mg QE/g). The HPLC analysis of date pits’ extract showed the presence of six phenolic compounds. Interestingly, there was a significant amount of gallic acid (11.85 mg GA/gFW), which was isolated via chromatography (thin layer chromatography) and identified by spectroscopic investigation (FTIR, MS, 1H, and 13C NMR). An inhibitory effect on Gram-positive and Gram-negative bacteria was observed for date pit gallic acid purified. S. aureus cell viability was completely inhibited at 200 µg/mL of acid for 6 h. Moreover, gallic acid was found to have a significant antimutagenic activity against Salmonella typhimurium at all tested concentrations, with a decrease in percent mutagenicity from 52 to 32 in the case of test substance TA98, and from 39 to 15 for test substance TA100 in terms of percent antimutagenicity. Gallic acid at a dose of 3000 g/plate had the greatest antimutagenic effect on S. typhimurium TA98, with the lowest toxicity. Overall, the bioactive properties of date pits’ extracts were investigated, with a particular attention to their chemical composition, biological activity, and pharmaceutical applications.

High Dilution of Dexamethasone in gestation and fetal development of mice

Background: Recently, the use of homeopathy in veterinary medicine has grown significantly, mainly for farm animal practice, because of its usefulness in organic production and low cost. There is a  wide range of veterinary products available in the  marketoften used in females. However, the effect of these products in the litter and derived products for human consummation is completely unknown. Aims: this  study sought to  develop an experimental model to study the putative effects of high diluted substances in newborns after chronic exposure of females. Methods: based on previous studies, the chosen test substance was dexamethasone 15cH; adult female Balb/c mice were divided into 4 groups: a) treated with PBS (control); b) treated with dexamethasone 15 cH; c) treated with dexamethasone 15cH + dexamethasone 4 mg/kg and d) treated with dexamethasone 4 mg/kg. All medicines were administered subcutaneously, 3 times a week, in females from the first day of pregnancy up to the 20th day after parturition (end of lactation period). TDevelopment of the offspring was evaluated daily  for 15 days after birth. Parameters evaluated were: female and offspring viability, number of newborns, time for eye opening, pinna opening, fur growth and postural reflex. Results: the group treated with dexamethasone 15cH  showed 39% increase in mortality rate 39 days after the beginning of treatment and 35% increase in fetal mortality at the end of gestation (p=0.0049). Females treated with dexamethasone 4mg/kg + dexamethasone 15cH showed 100% of fetal mortality. After parturition newborn survival in animals exposed to dexamethasone 4 mg/kg was higher than the control (p=0.0002). All other parameters of neonatal development were unchanged among groups. Conclusions: these data point to adverse effect when using high diluted dexamethasone during gestation detectable by this experimental model in Balb/c mice.

Comparative efficacy of transdermal forms for alopecia therapy

Introduction. Alopecia is a polyetiological disorder characterized by hair loss and reducing their number per unit area. Baldness causes psychological and social discomfort to patients, in connection with what an important task is to develop formulations that are more effective than the reference agents.Aim. Investigate the possibility of applying the original substance Y in several dosage forms for the treatment of alopecia in comparison with reference drugs: minoxidil and burdock oil.Materials and methods. The research subject was the original substance Y, for which several dosage forms were made: gel, alcohol and oil compositions. The study on the effectiveness and safety of the developed formulations was carried out on 9 groups of male C57BL/6 mice. Depilation with further assessment of the percentage of hair follicles in the growth and resting phases was tested as a pre-clinical model of alopecia. In the study of the mechanism of action of substance Y, chemiluminescent assay was performed compared with natural antioxidant quercetin in the system luminol – 2,2'-azo-bis(2-amidinopropane)dihydrochloride, in potassium-phosphate buffer medium (pH = 7.4). Statistical processing of the results was carried out using two-way ANOVA using GraphPad Prism 8.0.2, USA software at the level of statistical significance of differences p < 0.05 and p < 0.005.Results and discussion. Based on the results of histological analysis and visual changes, it was found that the effectiveness of the topical forms of substance Y decreases in the following order: gel, alcohol form, oil composition. The use of a combination of the gel base with the test substance Y resulted to the appearance of a larger number of hair follicles in the growth phase than when using the reference preparation – 2 % minoxidil solution (the differences are statistically significant). Chemiluminescent assessment of antioxidant activity showed the lack of antioxidant effect in substance Y.Conclusion. The study combines two pharmaceutical profiles: technological and pharmacological. In the course of the experiments, the prospects of the gel form of the original substance Y for topical therapy of alopecia were shown. In the near future, it is planned to study the mechanism of action of substance Y, as well as registration of patent protection for a new drug.

Reproducibility of effects of the homeopathic dilutions 14x - 30x of gibberellic acid on growth of Lemna gibba L.

Background: Reproducibility of investigations in homeopathy is still challenging. Duckweed (Lemna gibba L.), a monocotyledonous waterplant which mostly reproduces vegetatively and therefore builds genetically identical clones, may be a suitable test system for standardised trials. Aims: This study investigated if formerly observed effects of gibberellic acid 14x – 30x on growth of Lemna gibba were reproducible. Methododology: Duckweed was grown in dilutions of gibberellic acid (14x–30x) as well as once succussed (c1) and unsuccussed (c0) water control. Area-related growth rate for day 0–7 was determined by a computerised image analysis system. Three series including five independent blinded and randomised experiments each were carried out in the same way as in the original study. Only time and conductor of experiments were modified. System stability was controled by three series of systematic negative control (SNC) experiments with the same set-up, but distilled and autoclaved water was used as the only test substance. According to the series with gibberellic acid, each serie of SNC experiments included five experiments. Full two-way ANOVA (α = 5%) was used for statistical analysis. Independent variables were treatment and experiment number, dependent variable was r(area) for day 0–7. Data of each experiment was normalised to its mean value to allow a better comparison between experiments. Only if the global ANOVA F-test was significant (p < 0.05) we compared the investigated groups with Fisher`s LSD test (protected Fisher`s LSD). Results: No specific effects of agitated dilutions of gibberellic acid were found in the first two replication series (p=0.263 and p=0.062). In the third serie with gibbous Lemna gibba L. we observed a significant effect (p=0.009) of the homeopathic treatment, however growth was increased in contrast to decreasing in the former study. Variability in experiments with gibberellic acid 14x – 30x was lower than in SNC experiments. The stability of the experimental system was verified by the SNC experiments. Conclusions: When designing new studies to investigate reproducibility, different physiological states of the test organism must be considered. Variability might be an interesting parameter to investigate effects of homeopathic remedies in basic research.

High-Throughput in vitro Gel-Based Plate Assay to Screen for Calcium Oxalate Stone Inhibitors

<b><i>Objective:</i></b> Kidney stones are a common medical condition that is increasing in prevalence worldwide. Approximately, ∼80% of urinary calculi are composed of calcium oxalate (CaOx). There is a growing interest toward identifying therapeutic compounds that can inhibit the formation of CaOx crystals. However, some chemicals (e.g., antibiotics and bacterial metabolites) may directly promote crystallization. Current knowledge is limited regarding crystal promoters and inhibitors. Thus, we have developed an in vitro gel-based diffusion model to screen for substances that directly influence CaOx crystal formation. <b><i>Materials and Methods:</i></b> We used double diffusion of sodium oxalate and calcium chloride-loaded paper disks along an agar medium to facilitate the controlled formation of monohydrate and dihydrate CaOx crystals. A third disk was used for the perpendicular diffusion of a test substance to assess its influence on CaOx crystal formation. <b><i>Results:</i></b> We confirmed that citrates and magnesium are effective inhibitors of CaOx crystals. We also demonstrated that 2 strains of uropathogenic <i>Escherichia coli</i> are able to promote crystal formation. While the other tested uropathogens and most antibiotics did not change crystal formation, ampicillin was able to reduce crystallization. <b><i>Conclusion:</i></b> We have developed an inexpensive and high-throughput model to evaluate substances that influence CaOx crystallization.

Optimization of operating modes of the weak absorption spectrometer

The possibilities of improving the characteristics of a weak absorption spectrometer consisting of a frequency-tunable laser and an external analytical resonator with the test substance are analyzed. The influence of the scanning speed of the laser frequency on the choice of the spectrometer operating modes that provide the required resolution and the required sensitivity of spectral measurements is considered. Particular attention is paid to assessing the role of the interaction of modes in an analytical cavity on the structure of the recorded spectra. It was found that at a high rate of change in the laser frequency and superposition of fields of longitudinal modes, an improvement in the resolving power is combined with a certain decrease in the sensitivity and accuracy of recording weak spectral lines. It is shown that the optimization of the spectrometer operation modes requires correct accounting of the Q-factor of the analytical resonator and the line width of the probe radiation.

UJI AKTIVITAS ANTIBAKTERI ISOLAT SENYAWA GOLONGAN TERPENOID DARI BIJI PEPAYA (Carica papaya L.) TERHADAP Staphylococcus aureus SECARA IN VIVO PADA KELINCI JANTAN

research on the isolation of terpenoid class of compounds from the seeds of papaya (Carica papaya L.) and test its activity against Staphylococcus aureus in vivo in male rabbits. This research aims to prove that the terpenoid compounds isolated from the seeds of papaya (Carica papaya L.) can inhibit the growth of Staphylococcus aureus in vivo. Separation of terpenoid compounds by column chromatography ((eluent n-hexane: ethyl acetate: ethanol)) resulted in 25 eluates, and then merged based on the results of identification by TLC ((nhexane: ethyl acetate (8: 2)) and the color test reagent Lieburmann -Burchard produce 5 fraction groups. fraction D showed positive terpenoids with Rf 0.75 and the color purple with Lieburmann-Burchard reagent. study using 15 rabbits were divided into 5 groups: P1 (negative control), P2 (positive control) , P3 (5% of the test group), P4 (10% of the test group), P5 (test group 15%). each group was given the intracutaneous Staphylococcus aureus as 0,2ml on the backs of rabbits. Having symptoms of infection each group was given the test substance 3 times a day topically, the observed parameter is the diameter of the wound, and histopathological observations performed on days 3,6 and 9 Analysis of the results of research conducted using ANSIRA showed highly significant differences between groups (p <0.05). Then proceed with the analysis of the results of the analysis HSD test showed highly significant differences in the test group 5% to 10% of the test group and the test group 15%. Isolates terpenoid class of compounds from the seeds of papaya (Carica Papaya L.) with a concentration of 10% and 15% can inhibit the growth of Staphylococcus aureus. Keywords: Antibacterial, Staphylococcus aureus, male rabbits

Virucidal activity of Nasaleze Cold® & Flu Blocker and Nasaleze® Travel in cell cultures infected with human pathogenic coronavirus 229-E

This in vitro study determined the anti-viral efficacy of a unique blend of powder cellulose supplemented with powdered garlic extract (PGE) and a signalling agent. The composition, presented as Nasaleze Cold & Flu Blocker/Nasaleze Travel, was assessed against Human Coronavirus 229E, CoV 229E {ATCC VR-740} in an in vitro experiment. The test substance was used at sub-optimal dosing levels to explore its prevention and treatment capabilities. The virucidal activity of this novel formulation was measured at 48, 72 and 112 hour periods after incubation. Results showed strong reductions in viral titre of Coronavirus 229E compared to a control, while no toxicity to human cells from the test formulation was noted. The extract Nasaleze Cold/Travel showed potential to be used as a therapeutic and preventive agent. The data reconfirms the established anti-viral activity of this formulation acting as a barrier preventing the virus from accessing the nasal mucosa and disrupting its replication.