Chromatographic Profiles Research Articles

First Insights into the Venom Composition of Two Ecuadorian Coral Snakes.

Micrurus is a medically relevant genus of venomous snakes composed of 85 species. Bites caused by coral snakes are rare, but they are usually associated with very severe and life-threatening clinical manifestations. Ecuador is a highly biodiverse country with a complex natural environment, which is home to approximately 20% of identified Micrurus species. Additionally, it is on the list of Latin American countries with the highest number of snakebites. However, there is no local antivenom available against the Ecuadorian snake venoms, and the biochemistry of these venoms has been poorly explored. Only a limited number of samples collected in the country from the Viperidae family were recently characterised. Therefore, this study addressed the compositional patterns of two coral snake venoms from Ecuador, M. helleri and M. mipartitus, using venomics strategies, integrating sample fractionation, gel electrophoresis, and mass spectrometry. Chromatographic and electrophoretic profiles of these snake venoms revealed interspecific variability, which was ascertained by mass spectrometry. The two venoms followed the recently recognised dichotomic toxin expression trends displayed by Micrurus species: M. helleri venom contains a high proportion (72%) of phospholipase A2, whereas M. mipartitus venom is dominated by three-finger toxins (63%). A few additional protein families were also detected in these venoms. Overall, these results provide the first comprehensive views on the composition of two Ecuadorian coral snake venoms and expand the knowledge of Micrurus venom phenotypes. These findings open novel perspectives to further research the functional aspects of these biological cocktails of PLA2s and 3FTxs and stress the need for the preclinical evaluation of the currently used antivenoms for therapeutic purposes in Ecuador.

Process Development of a Triphasic Continuous Flow Suzuki–Miyaura Coupling Reaction in a Plug Flow Reactor

We have successfully adapted a Suzuki–Miyaura coupling reaction to a continuous flow process. The reaction was a triphasic mixture involved with two liquid phases that are only partially miscible in addition to the presence of an insoluble catalyst as a solid phase. The reaction required very rapid reaction heat up and mixing, which were difficult to achieve under batch conditions. A plug flow reactor (PFR) equipped with a static mixer was implemented in this continuous flow process, and the heterogenous reaction mixture was well handled, which is attributed to the high velocity flow rates and fast reaction kinetics. The reaction was successfully carried out on a 1.6 kg scale to give 80% isolated yield of the product with superior reaction high-performance liquid chromatography profile to the batch process.

Physico Chemical Analysis and HPTLC Evaluation of Kokilakshadi Kwatha Churna

Kokilakshadi Kashaya mentioned in Bhaishajya Ratnavali, Vataraktadhikara, it is a therapeutic formulation to treat Vatarakta. It is also used by Ayurvedic practitioners for treating hyperuricemia. The symptoms of hyperuricemia and gouty arthritis are similar to Vatarakta, a disease explained in classical Ayurvedic textbooks. Kokilakshadikwatha contains Kokilaksha and Guduchi and Pippalichurna given as Anupana of this formulation Physico chemical analysis of individual drug and formulation with modern parameters increase their scope and acceptance. The study was based on standard analytical parameters proposed by API. Method: Kokilakshadi Kwatha powder was evaluated for physico chemical analysis and phyto chemical screening. The analysis was done by using the parameters like Organoleptic features, loss on drying, acid soluble extractive, water soluble extractive. Results: Analytical parameters of individual drugs were done. All analytical parameter were within limit. Analytical parameter of Kokilakshadi Kwatha Churna like loss on drying 10.4%w/w, acid insoluble ash 0.79%, alcohol soluble extractive 11.2%w/w, water soluble extractive 7.8%w/w, pH 5.78 were obtained. High Performance Thin Layer Chromatography (HPTLC) profile of Kokilakshadikwatha powder showed 13 peaks at 254nm and 14 peaks at 366nm. Preliminary phytochemical screening test revealed the presence of steroids, alkaloids, phenols, flavonoids. Conclusion: The obtained data can be used for future comparative references

Iron Content, Iron Speciation and Phycocyanin in Commercial Samples of Arthrospira spp.

Cyanobacteria are characterized by high iron content. In this research, we collected ten commercial samples of Arthrospira spp. sold as food supplement to determine iron content and assess whether iron speciation showed variability among samples and changed respect to A. platensis grown in controlled conditions. Particular attention was also paid to phycocyanin, as an iron-binding protein. In six of the ten samples, 14 essential and non-essential trace elements were analysed using ICP-MS. Iron content measured in samples using atomic absorption spectrometry (AAS) varied from 353 (sample S5) to 1459 (sample S7) µg g-1 dry weight and was in the range of those reported by other authors in commercial supplements. Iron speciation was studied using size exclusion chromatography followed by the analysis of the collected fraction for the determination of iron by AAS and for protein separation using SDS-PAGE. Overlapping chromatographic profiles were obtained for total proteins, phycocyanin and iron, although quantitative differences were evidenced among the samples analysed. In most samples, iron was mainly bound to ligands with high molecular mass; however, in four samples iron was also bound to ligands with low molecular mass. In fractions containing the most relevant iron burden, the principal protein was phycocyanin, confirming its role as an iron-binding protein in commercial samples.

The role of dietary patterns and erythrocyte membrane fatty acid patterns on mild cognitive impairment

Dietary fatty acids have been shown to be associated with the development of cognition. However, research on the role of fatty acid intake in dietary patterns and fatty acid patterns (FAPs) in the development of cognitive function is limited. The aim of this study was to explore the correlation between dietary patterns and FAPs and to provide available evidence for preventing mild cognitive impairment (MCI) through these patterns. The 973 participants aged between 65 and 85 were recruited from 2020 to 2021 for this multicenter research in Beijing. Neuropsychological tests were used for cognitive evaluation, and data of dietary intake in the past 12 months were collected with semi-quantitative food frequency questionnaire. The erythrocyte membrane fatty acid profile was tested by chromatography and mass spectrometry lipid profiling. Factor analysis was used to derive the main dietary patterns and FAPs. Pearson's correlation or Spearman's correlation was used to explore the association between dietary patterns and FAPs. Binary logistic regression was applied to examine the relationship between patterns and cognitive function. Six dietary patterns and six FAPs were identified, explaining 53.4 and 80.9% of the total variance separately. After adjusting all potential confounders, T3 of the pattern 1 and FAP2 were the independent protect factors for MCI, respectively (OR 0.601, 95% CI [0.395, 0.914]; OR 0.108, 95% CI [0.019, 0.623]). Rich of SM (26:0), SM (24:1), and SM (26:1) is the characteristic of FAP2. A positive correlation was found between component scores of dietary pattern1 and FAP2 (r = 0.441, p = 0.001). People who adhered to a reasonable intake of animal flesh consumed more various long-chain fatty acids as well. The erythrocyte membrane metabolites, SM (26:0), SM (24:1), and SM (26:1), might function as early biomarkers for predicting or monitoring of cognitive aging in the elderly. The dietary pattern with recommended animal flesh consumption was significantly associated with FAP characterized by very long-chain SMs. This dietary pattern affected FAP, which might achieve the ultimate goal of neuroprotection through the very long-chain SMs. A rational intake of dietary fatty acids might be an effective way on preventing MCI in the elderly.

STANDARDIZATION OF SECONDARY METABOLITES AND HEAVY METAL CONTAMINATION ASSAY ON ONCHIDIID SLUG (ONCHIDIUM TYPHAE) WEST KALIMANTAN WATERS

Onchidiid slug or Onchidium typhae is a marine animal commonly found in West Kalimantan waters. This gastropod is commonly used as medicine and cosmetic by the locals. This research aims to assure the content and quality of O. typhae methanol extract by determining its secondary metabolites and heavy metal contamination. Phytochemical screening of alkaloid, steroid/ triterpenoid, flavonoid, tannin, and saponin are done using reagent tests. Chromatography profile was completed using silica gel G60 F254 plate and hexane: ethyl acetate: methanol as mobile phase. Heavy metal contamination analysis was performed using atomic absorption spectrophotometry to determine Hg, As, Cd, and Pb content. The phytochemical screening and chromatography profile of O. typhae methanol extract confirm the presence of alkaloids and steroids. Heavy metal analysis stated Hg, As, Cd, and Pb content in O. typhae are < 0,0008; < 0,001; < 0,001; and 0,05 ppm respectively. Since heavy metal levels are below the maximum contamination limit, it qualifies as a natural product ingredient.

AKTIVITAS ANTIOKSIDAN EKSTRAK ETANOL, METANOL, DAN AQUADES DARI KULIT BUAH MUNDAR (Garcinia forbesii)

Mundar (Garcinia forbesii) is a typical plant from South Kalimantan. Mundar pericaps can be used as a natural antioxidant. The use of solvents in the extraction process can affect the antioxidant activity. The purpose of this study was to determine of phytochemical compounds and antioxidant activity of the ethanol, methanol, and aquades extracts from pericaps of mundar (Garcinia forbesii). Mundar pericaps is dried, then made into powder. Mundar pericaps powder was extracted with three different solvents, ethanol 70%, methanol, and aquades. Phytochemical screening and analysis by thin layer chromatography were performed on the three extracts. The three extracts were tested for antioxidant activity by DPPH method. The results showed that ethanol and methanol extracts contained phytochemical compounds of phenolic, flavonoids, tannins, alkaloids, steroids, and terpenoids. Aquades extract contains phenolic, flavonoids, tannins, and alkaloids. The chromatographic profile showed that the ethanol and methanol extracts had an Rf value of 0.307. The antioxidant activity from highest to lowest was ethanol extract (192.64 ppm), methanol extract (236.64 ppm), and aquades extract (443.13 ppm) from mundar pericaps

Phytochemical profile and anthelmintic effects of Laurus nobilis essential oil against the ovine nematode Haemonchus contortus and the murine helminth model Heligmosomoides polygyrus

Small ruminant production in tropical and temperate countries faced substantial anthelmintic resistance due to the intensive use of commercial anthelmintic drugs. Therefore, alternative treatments including natural bioactive compounds with anthelmintic potential have been investigated looking for its successfully use in the parasite control. In the present study, we describe the chemical profile of Laurus nobilis essential oil (EO), the in vitro anthelmintic activity of L. nobilis EO against Haemonchus contortus and its in vivo anthelmintic effect against the murine helminth parasite model Heligmosomoides polygyrus. Chromatographic profile of L. nobilis (EO) extracted from the leaves of L. nobilis have shown the presence of monterpens 1,8-cineol (Eucalyptol) (29.47%), D-Limonène (18.51%) and Linalool (10.84%) in high fractions. The in vitro anthelmintic potential was expressed by an ovicidal effect against H. contortus egg hatching with inhibition value of 1.72 mg/mL and 87.5% of immobility of adult worms after 8 h of exposure to 4 mg/mL of L. nobilis EO. Regarding, the in vivo anthelmintic potential, L. nobilis (EO) at 2400 mg/kg bw completely eliminated the egg output of H. polygyrus after 7 days of oral treatment, together with a 79.2% of reduction in total worm counts. Based on the obtained results, L. nobilis EO showed promising in vitro and in vivo anthelmintic capacities against gastrointestinal parasites.

Physicochemical Profile, Antioxidant and Antimicrobial Activities of Honeys Produced in Minas Gerais (Brazil).

Honeys can be classified as polyfloral or monofloral and have been extensively studied due to an increased interest in their consumption. There is concern with the correct identification of their flowering, the use of analyses that guarantee their physicochemical quality and the quantification of some compounds such as phenolics, to determine their antioxidant and antimicrobial action. This study aims at botanical identification, physicochemical analyses, and the determination of total polyphenols, chromatographic profile and antiradical and antimicrobial activity of honey from different regions of Minas Gerais. Seven different samples were analyzed for the presence of pollen, and color determination. The physicochemical analyses performed were total acidity, moisture, HMF, reducing sugar, and apparent sucrose. The compound profile was determined by UHPLC/MS, the determination of total phenolics and antiradical activity (DPPH method) were performed by spectrophotometry, and minimum inhibitory and bacterial concentrations were determined for cariogenic bacteria. All honey samples met the quality standards required by international legislation, twenty compounds were detected as the main ones, the polyfloral honey was the only honey that inhibited all of the bacteria tested. Sample M6 (Coffee) was the one with the highest amount of total polyphenols, while the lowest was M4 (Cipó-uva). Regarding the antioxidant activity, M5 (Velame) had the best result and M4 (Cipó-uva) was the one that least inhibited oxidation. Of the polyfloral honeys, there was not as high a concentration of phenolic compounds as in the others. Coffee, Aroeira, Velame and Polyfloral have the best anti-radical actions. Betônica, Aroeira, Cipó-uva and Pequi inhibited only some bacteria. The best bacterial inhibition results are from Polyfloral.

Development of a novel, label-free N-glycan method using charged aerosol detection

Monoclonal antibodies (mAbs) are complex glycoproteins that are developed for treatment of various therapeutic indications such as cancer and autoimmune diseases. MAbs are glycosylated at conserved asparagine residues (N-X-S/T) of the Fc region at amino acid position 297 of the heavy chain. Glycans are important in governing the functions of efficacy and serum half-life of protein therapeutics and are part of the critical quality attribute panel for release testing. Traditionally, N-linked glycans are released from glycoproteins after denaturation and enzymatic digestion with PNGase F, followed by fluorescent labeling of the liberated glycans. The labeled glycans are then separated using hydrophilic liquid chromatography (HILIC) with fluorescence detection to generate chromatographic profile. Despite decades of use, this strenuous process remains unchanged, utilizing toxic reagents and extended sample preparation time. As an intervention, this report showcases a novel, label-free approach to detect and quantify N-glycans without using fluorescent labeling. Separation of glycans using mixed-mode PGC column along with detection of non-derivatized glycans using charged aerosol detector, the overall turnaround time can be greatly reduced with significant cost savings. The label-free method provides similar quantitative results as the conventional fluorescent labeled method, confirming the validity of the method for product release.

Chemotaxonomic Markers for the Leaf Buds of Common Finnish Trees and Shrubs: A Rapid UHPLC MS Fingerprinting Tool for Species Identification.

In this study, a chemotaxonomic tool was created on the basis of ultra-high-performance liquid chromatography–mass spectrometry (UHPLC–MS) for the identification of 13 common Finnish deciduous trees and shrubs from their leaf bud metabolites. The bud extracts were screened with UHPLC–ESI–QqQ–MS and UHPLC–ESI–Q–Orbitrap–MS to discover suitable markers for each species. Two approaches were tested in the marker selection: (1) unique species-specific markers to obtain selective fingerprints per species and (2) major markers to maximise the sensitivity of the fingerprints. The markers were used to create two selected ion-recording-based fingerprinting tools with UHPLC–ESI–QqQ–MS. The methods were evaluated for their selectivity, repeatability, and robustness in plant species identification by analysing leaf buds from several replicates of each species. The created chemotaxonomic tools were shown to provide unique chromatographic profiles for the studied species in less than 6 min. A variety of plant metabolites, such as flavonoids, triterpenoids, and hydroxycinnamic acid derivatives, were found to serve as good chemotaxonomic markers for the studied species. In 10 out of 13 cases, species-specific markers were superior in creating selective and repeatable fingerprints.

FcγRIIIA affinity chromatography complements conventional functional characterization of rituximab.

Analytical and functional characterization of batches of biologics/biosimilar products are imperative towards qualifying them for pre-clinical and clinical investigations. Several orthogonal strategies are employed to characterize the functional attributes of these drugs. However, the use of conventional techniques for online monitoring of functional attributes is not feasible. Liquid chromatography is one of the crucial unit operations during the downstream processing of biopharmaceuticals. In this work, we have demonstrated the utility of FcγRIIIA affinity chromatography as an independent quantitative functional characterization tool. FcγRIIIA affinity chromatography aided in sequential elution of Rituximab glycoform mixtures, based on varying levels of galactosylation, and thereby the affinity for the receptor protein. The predominant glycans present in the three Rituximab glycoform mixture peaks were G0F, G1F, and G2F, respectively. Dissociation rate constants were derived from the chromatographic elution profiles by the peak profiling method, for the control and glucose stress conditions. The glucose stress conditions did not result in unfavorable binding kinetics of Rituximab and FcγRIIIA. The dissociation rate constants of the glycoform mixture 2, predominantly consisting of G1F, were similar to the dissociation rate constants obtained by surface plasmon resonance. Moreover, the glycosylation profiles obtained from chromatographic estimation can be corroborated with the ADCC activity. However, the ex vivo ADCC reporter assay indicated that there was an increase in the effector activity with increasing glucose stress. Thus, FcγRIIIA affinity chromatography permitted three independent assessments via a single analysis. Such approaches can be utilized as potential process analytical technology (PAT) tools in the biosimilar development process.

Hydrobiological and Bio-Optical Characterization of a Red Tide Occurredin Meda Creek, Porbandar, India

Red tides are becoming more common in aquatic ecosystems, particularly the coastal habitats which are highly influenced by climatic and anthropogenic activities. We provide the first report of a red tide event in Meda creek, west coast of India. Light and scanning electron microscopy were used to identify the bloom forming organism, while physicochemical, biological and biooptical parameters were determined to characterize the aquatic body of the Meda Creek. The dominant bloom forming organisms were observed as unarmored, spherical, dorsoventrally compressed flagellate with truncated apex, notched and rounded hypocone, displaced premedian cingulum and extending sulcus. The length wise size of the cell varied from 20 to 30.83μM, whereas, the width varied from 14 to 27.5μM. The organisms was identified as Gymnodinium sp. High Performance Liquid Chromatography (HPLC) and spectral profile of the bloom indicatedeleven major peaks corresponding to that of 19-but-fucoxanthin (52.75%), astaxanthin (9.62%), fucoxanthin (8.81%), Mgdv (6.45%), prasinoxanthin (3.35%), chlorophyll C2 (3.22%), neoxanthin (3.12%), antheraxanthin (3.14%), diadinoxanthin (2.63%) and diatoxanthin (1.38%). Chlorophyll a concentration in the bloom sample was 38.8μg/L. Water temperature, salinity, NH3-N, NO2-N, NO3-N, PO4-Pwere higher during the bloom period than the non-blooming ones, while metals including Fe, Zn, Cr, Mn and Co were lower. No death of aquatic biota was observed. We propose that eutrophication along with the warm and saline conditions provided enabling conditions for occurrence the bloom. Also, most of the pigments expressed have antioxidant role and might be purposeful for the survival of the organism in the extreme condition of temperature and salinity recorded. Proper management and monitoring measures might help mitigate further occurrences of this bloom in the light changing climate and increasing anthropogenic impacts.

Thin Layer Chromatography of Secondary Metabolite Profiles and Determination of Total Flavonoid Levels of the Sawi Langit (Vernonia Cinerea L.) Extract that Growing on Peatlands

Peatland is a type of soil with a high organic content, typically more than 65 percent organic matter, high acidity, and low nutrient content. Vernonia cinerea, also known as Sawi Langit in Indonesia, is one of the nutritious plants. The community uses this herb to treat fevers, wounds, discomfort, and ulcers. The secondary metabolite concentration of plants in peatlands is affected by nutrients such as carbon, nitrogen, and pH. Find out the secondary metabolite content of Sawi Langit (Vernonia cinerea L.) leaf extract and the total flavonoid levels that grown on peatlands. The Rf value was estimated after the descriptive observational approach was obtained from the Thin Layer Chromatography (TLC) profile of secondary metabolites with spray reagents, and the quantitative data was determined by UV-Vis spectrophotometry. Sawi Langit plants were found growing on peatlands with a pH of 3.77, N-total of 1.51%, and C-Organic of 4.16%. The best eluent for separating compounds is chloroform: methanol: water (2:5:3) for flavonoid compounds, ethyl acetate: methanol: water (16:1:2) for alkaloid compounds, chloroform: methanol (9:1) for steroid compounds, n-hexane: ethyl acetate (12:8) for terpenoid compounds, ethyl acetic: chloroform: acetic acid (15:5:2) for tannin compounds, and chloroform: ethanol: water (10:6:1) for saponin compounds. Sawi langit (Vernonia cinerea L.) leaf extract has a total flavonoid concentration of 5.527 mgQE (Quersetin Equivalent)/g. Flavonoid, alkaloids, steroids, terpenoids, tannins, and saponins compounds were found in the TLC profile of Sawi Langit (Vernonia cinerea L.) leaf extract, and the total flavonoid content of the leaf extract of Sawi Langit (Vernonia cinerea L.) was 5.527 mgQE (Quersetin Equivalent)/g.

ANALYSIS OF GENUS SPECIFIC PHENOLIC COMPOUNDS IN PLANTS OF THE GENUS RHODIOLA SPP. IN A COMPARATIVE ASPECT

Plants of the genus Rhodiola (Crasulaceae) are actively used in folk medicine around the world. The most studied representative of this genus is Rhodiola rosea L., which is an official species in the Russian Federation. But in the folk medicine of Russia and China, other species of the genus Rhodiola are also used. Thus, in the southern part of Eastern Siberia, Rhodiola quadrifida (Pall.) Fisch et Mey is very popular., in Central Asia - Rhodiola heterodonta (Hook. f. & Thomson) Boriss. To introduce these species of the genus Rhodiola into official medicine, it is necessary to determine marker compounds and select standardization criteria. The purpose of this work was to study phenolic alcohols and their glycosides, established as genus specific, in Rhodiola quadrifida and Rhodiola heterodonta in comparison with Rhodiola rosea by high performance thin layer and liquid chromatography. It has been established that rosarin is characteristic only for R. rosea, while salidroside and tyrosol quantitatively prevail in R. heterodonta and R. quadrifida. Based on the obtained results, it can be assumed that the differences in the chromatographic profile of the studied species can be used to confirm the identity, and the identified compounds can be used to standardize the indicated Rhodiola species.

High‐performance thin‐layer chromatography a densitometric detection of multi‐class bioactive compounds from three species of marine algae Padina and identification of antioxidant substance with mass spectrometry

AbstractPadina is rolled‐blade‐shaped marine brown algae that often have overgrown and has been well explored for its medicinal qualities. The various species of Padina release a wide range of promising chemical components as part of their chemical defense systems. In this context, a finding was conducted to explore the bioactive compounds using exceptionally sophisticated analytical techniques, such as high‐performance thin‐layer chromatography. Microchemical and biochemical derivatization was used in the investigation to confirm multi‐class bioactive molecules and the antioxidant activity of the separated band. Microchemical derivatization ascertained the appearance of miscellaneous lipophilic substances, essential oils, volatile compounds, coumarin, and bitter principles. To determine the chemical standard of these algae, the qualitative (microchemical) screening of phytochemicals associated with flexible high‐performance thin‐layer chromatography was utilized. Moreover, biochemical derivatization, which generates a spectacular white band with a purple background to appear on the chromatogram, substantiates the antioxidant activity of a particular band. These results proffer a chromatographic profile of the antioxidant capability of three species of Padina's bioactive components, which will create the necessary information for the standardization and identification of marine algae. Mass spectrometry results revealed the presence of phytol compound from the diterpenoid class, which has antioxidant potential.

Bactericidal Activity of Carvacrol against Streptococcus pyogenes Involves Alteration of Membrane Fluidity and Integrity through Interaction with Membrane Phospholipids

Background: Carvacrol, a mono-terpenoid phenol found in herbs, such as oregano and thyme, has excellent antibacterial properties against Streptococcus pyogenes. However, its mechanism of bactericidal activity on S. pyogenes has not been elucidated. Objectives: This study investigated the bactericidal mechanism of carvacrol using three strains of S. pyogenes. Methods: Flow cytometry (FCM) experiments were conducted to determine carvacrol’s membrane permeabilization and cytoplasmic membrane depolarization activities. Protoplasts of S. pyogenes were used to investigate carvacrol’s effects on the membrane, followed by gel electrophoresis. The carvacrol-treated protoplasts were examined by transmission electron microscopy (TEM) to observe ultrastructural morphological changes. The fluidity of the cell membrane was measured by steady-state fluorescence anisotropy. Thin-layer chromatographic (TLC) profiling was conducted to study the affinity of carvacrol for membrane phospholipids. Results: Increased membrane permeability and decreased membrane potential from FCM and electron microscopy observations revealed that carvacrol killed the bacteria primarily by disrupting membrane integrity, leading to whole-cell lysis. Ultra-structural morphological changes in the membrane induced by carvacrol over a short period were confirmed using the S. pyogenes protoplast and membrane isolate models in vitro. In addition, changes in the other biophysical properties of the bacterial membrane, including concentration- and time-dependent increased fluidity, were observed. TLC experiments showed that carvacrol preferentially interacts with membrane phosphatidylglycerol (P.G.), phosphatidylethanolamine (P.E.), and cardiolipins (CL). Conclusions: Carvacrol exhibited rapid bactericidal action against S. pyogenes by disrupting the bacterial membrane and increasing permeability, possibly due to affinity with specific membrane phospholipids, such as P.E., P.G., and CL. Therefore, the bactericidal concentration of carvacrol (250 µg/mL) could be used to develop safe and efficacious natural health products for managing streptococcal pharyngitis or therapeutic applications.

Biophysical properties of human body louse nit related proteins: LNSP1, Agp9 and Agp22

The human parasitic head and body lice lay their eggs on either hair or clothing. Attachments of the eggs are possible because the female lice secret a glue substance from the accessory gland along with the egg, which hardens into a nit sheath that secures and protects the egg (The “nit” commonly refers to either the louse egg with an embryo or the empty hatched egg). Proteins called the louse nit sheath protein (LNSP) are suggested to be the major proteins of the nit sheath, but transcriptome profiling of the accessory glands indicated other proteins such as Agp9 and Agp22 are also expressed in the glands. In this study, human body louse LNSP1 (partial), Agp9, and Agp22 are recombinantly produced using the E. coli expression system, and the biophysical properties characterized. Circular dichroism analysis indicated that the secondary structure elements of LNSP1 N-terminal and middle-domains, Agp9, and Agp22 are prominently random coiled with up to 10–30% anti-parallel β-sheet element present. Size-exclusion chromatography profiles of LNSP1 proteins further suggested that the β-sheets made of the smaller N-terminal domain stacks onto the β-sheets of the larger middle-domain.

In Vitro Bioactivities of Extracts from Tomato Pomace.

Tomato pomace (TP) is a coproduct generated by the extraction of tomato pulp, and is a potential source of bioactive molecules. In this study, we isolated several fractions from TP and evaluated their biological properties. TP was treated by maceration at room temperature with green solvents (ethanol, ethyl acetate, ethanol:water and ethanol:ethyl acetate) or supercritical CO2 (SC-CO2). The extracts were analyzed by HPLC-DAD to determine their composition, and their antioxidant activity was assessed. The potential therapeutic effects of the isolated fractions were assessed in vitro. We identified 30 molecules on chromatography profiles, which revealed an abundance in phenolic acids, carotenoids, flavonoids and tannins, with differences in selectivity according to the solvent and pretreatment used. The highest radical scavenging activities were measured at 64-72% inhibition, corresponding to the ethanol or ethanol:water extracts with the highest polyphenol or flavonoid contents. Carotenoid content was increased by chemical pretreatment, to attain levels of 161 mg β-carotene/g ethyl acetate extract. This level of carotenoids seemed to have anti-inflammatory effects, with an IC50 of 9.3 μg/mL. In terms of anti-diabetic effects, the activities of α-glucosidase and α-amylase were best inhibited by extraction in an ethanol-to-water mixture (50:50). Cytotoxicity in a tumor cell line were highest for SC-CO2 extracts (64.5% inhibition) and for ethanol extracts obtained after the enzymatic pretreatment of TP (37% inhibition). Some extracts also had dose-dependent activity against Zika virus. New fractions obtained from TP with ecocompatible solvents in mild conditions are rich in bioactive molecules. A comparison of the chromatographic profiles of the extracts led to the identification of several key molecules with therapeutic properties. The chemical pretreatment of TP is justified as a mean of increasing the carotenoid content of ethyl acetate fractions, whereas enzymatic pretreatment can increase the antioxidant activity of ethyl acetate fractions and increase the cytotoxicity of ethanol fractions. The SC-CO2 fraction contained a smaller number of metabolites detectable on HPLC, but it had high levels of cytotoxicity and antioxidant activity. Finally, the fractions obtained appeared to be suitable for use to target one or several of the biological activities studied.

<em>Ocimum campechianum</em> essential oil: chemical composition and antifungal activity against filamentous fungi

Ocimum campechianum (OC), an aromatic and medicinal plant from the Amazon region, is rich in essential oils with antimicrobial potential. This study aimed to analyze the quality, toxicity, and antifungal action of Ocimum campechianum essential oil(OCEO). The plant species were collected in the city of São Luís-MA (Brazil). The essential oil (EO) was extracted by hydrodistillation in a Clevenger apparatus, and the physical-chemical parameters were analyzed according to the Brazilian Pharmacopoeia norms. The essential oil constituents were identified by gas chromatography-mass spectrometry (GC-MS). Toxicity was evaluated against brine shrimp (Artemia salina Leach). The antifungal activity was evaluated by the method of disk diffusion and microdilution in broth against ATCC strains of filamentous fungi Aspergillus Niger (ATCC 6275), Colletotrichum gloeosporioides (ATCC 96723), and Penicillium chrysogenum (ATCC 10106. The essential oil showed a slightly yellowish color, intense clove aroma, a density of 1.0 ± 0.0 g/mL, a refractive index of 1.52 ± 0.00, solubility in ethanol (1:2), and a yield of 1.40%. The chromatographic profile showed 36 volatile compounds, the majority being eugenol (49.18%), followed by β- caryophyllene (17.26%), elixene (9.46%), methyl eugenol (7.65%), and eucalyptol (2,55%). It presented LC 50 = 955 μg / mL against Artemia salina. The fungi induced growth inhibition zones (14.90 to 17.85 mm). The fungus P. chrysogenum was the most susceptible (MIC 220 μg/ml) compared to the fungi A. niger (MIC 250 μg/mL) and C. gloeosporioides (MIC 270 μg/mL). The minimum fungicide concentration (MFC) was sufficient to inhibit the growth of P. chrysogenum at 380 μg/mL, A. niger at 400.25 μg/mL, and C. gloeosporioides at 416.66 μg/mL. The OCEO showed good quality, low toxicity, and fungicidal action against the filamentous fungi analyzed, suggesting its application as a promising natural antifungal, mainly in combating diseases in post-harvest crops.