Screening Of Metabolites Research Articles

New-born screening of metabolites with liquid chromatography-mass spectrometry and gas chromatography–mass spectrometry: a metabolic and molecular signatures methyl malonic aciduria with combined oxidative phosphorylation deficiency-10: a rare case report

An uncommon autosomal recessive organic acid disease is malonic aciduria. This disease may be easily identified and included in the NBS programmes by means of the widespread use of tandem mass spectrometry’s in the study of the amino acid/acylcarnitine profile using dried blood spots (DBS) for newborn screening. In Tamil Nadu, we reported the first screened and diagnosed with malonic aciduria by newborn screening (NBS) in early neonatal period. The patient possesses a malonyl-CoA decarboxylase genetic variation not previously described. This disease should be distinguished from a related malonic and methylmalonic aciduria problem. Malonic aciduria's clinical phenomenology varies and pathogenesis is not completely known. The proper treatment regimen, nutritional therapy or frequent follow-up to these individuals are not guided or recommended. The majority of current treatment data is based on a single research or case report.

Pengembangan Buku Saku Berdasarkan Studi Identifikasi Tanaman Obat Pada Masyarakat Suku Serawai Bengkulu Selatan

This research is a research and development of teaching materials in the form of a pocketbook based on the study of the identification of medicinal plants in the Serawai people of South Bengkulu. The purpose of this study was to determine the medicinal plants, the content of secondary metabolite phytochemical compounds in each plant, and the validity and responses of students to the developed pocketbook. The development model used is the research and development (R&D) model of Bord and Gall (1983) which is limited to the seventh stage, namely the revision of the product from the limited trial results. From the research, 19 (nineteen) medicinal plants were selected that were used by the Serawai people of South Bengkulu. The results of secondary metabolite screening showed that the medicinal plants found contained various levels of tannins, alkaloids, saponins, flavonoids, terpenoids, and steroids. The validity of the pocketbook from the study of the identification of medicinal plants in the Serawai tribal community obtained an average value of 84.3% for the material aspect and 91.7% for the media aspect which was included in the very appropriate category. Student responses regarding the pocket book of medicinal plants developed obtained a value the percentage of material aspects, media, benefits are 85.8%, 88.0%, and 87.0%, respectively with an average of 86.9% which were included in the very good criteria. Student also give a positive response to the product being developed with a 100 % interest percentage.
 Keywords: Pocket Book, Medicinal Plants, Serawai Tribe, Secondary Metabolites

Detection and Quantification of Ammonia as the Ammonium Cation in Human Saliva by 1H NMR: A Promising Probe for Health Status Monitoring, with Special Reference to Cancer.

Ammonia (NH3) has been shown to be a key biomarker for a wide variety of diseases, such as hepatic and chronic kidney diseases (CKD), and cancers. It also has relevance to the oral health research area, and, hence, its determination in appropriate biofluids and tissues is of much importance. However, since it contains exchangeable >N-H protons, its analysis via 1H NMR spectroscopy, which is a widely employed technique in untargeted metabolomic studies, is rendered complicated. In this study, we focused on the 1H NMR analysis of this biomarker in less invasively collected human saliva samples, and we successfully identified and quantified it as ammonium cation (NH4+) in post-collection acidulated forms of this biofluid using both the standard calibration curve and standard addition method (SAM) approaches. For this purpose, n = 27 whole mouth saliva (WMS) samples were provided by healthy human participants, and all donors were required to follow a fasting/oral environment abstention period of 8 h prior to collection. Following acidification (pH 2.00), diluted WMS supernatant samples treated with 10% (v/v) D2O underwent 1H NMR analysis (600 MHz). The acquired results demonstrated that NH4+ can be reliably determined in these supernatants via integration of the central line of its characteristic 1:1:1 intensity triplet resonance (complete spectral range δ = 6.97-7.21 ppm). Experiments performed also demonstrated that any urease-catalysed NH3 generation occurring post-sampling in WMS samples did not affect the results acquired during the usual timespan of laboratory processing required prior to analysis. Further experiments demonstrated that oral mouth-rinsing episodes conducted prior to sample collection, as reported in previous studies, gave rise to major decreases in salivary NH4+ levels thereafter, which renormalised to only 50-60% of their basal control concentrations at the 180-min post-rinsing time point. Therefore, the WMS sample collection method employed significantly affected the absolute levels of this analyte. The LLOD was 60 μmol/L with 128 scans. The mean ± SD salivary NH4+ concentration of WMS supernatants was 11.4 ± 4.5 mmol/L. The potential extension of these analytical strategies to the screening of other metabolites with exchangeable 1H nuclei is discussed, as is their relevance to the monitoring of human disorders involving the excessive generation and/or uptake of cellular/tissue material, or altered homeostasis, in NH3.

CRISPR-Cas9 Gene Editing and Secondary Metabolite Screening Confirm Fusarium graminearum C16 Biosynthetic Gene Cluster Products as Decalin-Containing Diterpenoid Pyrones.

Fusarium graminearum is a causal organism of Fusarium head blight in cereals and maize. Although a few secondary metabolites produced by F. graminearum are considered disease virulence factors, many molecular products of biosynthetic gene clusters expressed by F. graminearum during infection and their associated role in the disease are unknown. In particular, the predicted meroterpenoid products of the biosynthetic gene cluster historically designated as "C16" are likely associated with pathogenicity. Presented here are the results of CRISPR-Cas9 gene-editing experiments disrupting the polyketide synthase and terpene synthase genes associated with the C16 biosynthetic gene cluster in F. graminearum. Culture medium screening experiments using transformant strains were profiled by UHPLC-HRMS and targeted MS2 experiments to confirm the associated secondary metabolite products of the C16 biosynthetic gene cluster as the decalin-containing diterpenoid pyrones, FDDP-D and FDDP-E. Both decalin-containing diterpenoid pyrones were confirmed to be produced in wheat heads challenged with F. graminearum in growth chamber trials. The extent to which the F. graminearum C16 biosynthetic gene cluster is dispersed within the genus Fusarium is discussed along with a proposed role of the FDDPs as pathogen virulence factors.

LC-MS profiling-based non-targeted secondary metabolite screening for deciphering cosmeceutical potential of Malaysian algae.

Application of natural resources from the marine environment in the cosmeceutical industry is gaining great attention. This study pursues to discover the cosmeceutical potential of two Malaysian algae, Sargassum sp. and Kappaphycus sp. by determining their antioxidant capacity and assessing the presence of their secondary metabolites with cosmeceutical potential using non-targeted metabolite profiling. Metabolite profiling using Quadrupole Time-of-Flight (Q-TOF) liquid chromatography-mass spectrometry (LC-MS) in the Electrospray Ionization (ESI) mode resulted in 110 putative metabolites in Sargassum sp. and 47 putative metabolites in Kappaphycus sp. and were grouped according to their functions. To the best of our knowledge, the bioactive compounds of both algae have not been studied in any great detail. This is the first report to explore their cosmeceutical potential. Six antioxidants were detected in Sargassum sp., including fucoxanthin, (3S, 4R, 3'R)-4-Hydroxyalloxanthin, enzacamene N-stearoyl valine, 2-hydroxy-hexadecanoic acid, and metalloporphyrins. Meanwhile, three antioxidants detected in Kappahycus sp., namely Tanacetol A, 2-fluoro palmitic acid and idebenone metabolites. Three antioxidants are found in both algae species, namely, 3-tert-Butyl-5-methylcatechol, (-)-isoamijiol, and (6S)-dehydrovomifoliol. Anti-inflammatory metabolites such as 5(R)-HETE, protoverine, phytosphingosine, 4,5-Leukotriene-A4, and 5Z-octadecenoic acid were also found in both species. Sargassum sp. possesses higher antioxidant capacity as compared to Kappahycus sp. which may be linked to its number of antioxidant compounds found through LC-MS. Hence, our results conclude that Malaysian Sargassum sp. and Kappaphycus sp. are potential natural cosmeceutical ingredients as we aim to produce algae cosmeceutical products using native algae.

Construction of the XGBoost model for early lung cancer prediction based on metabolic indices

BackgroundLung cancer is a malignant tumour, and early diagnosis has been shown to improve the survival rate of lung cancer patients. In this study, we assessed the use of plasma metabolites as biomarkers for lung cancer diagnosis. In this work, we used a novel interdisciplinary mechanism, applied for the first time to lung cancer, to detect biomarkers for early lung cancer diagnosis by combining metabolomics and machine learning approaches.ResultsIn total, 478 lung cancer patients and 370 subjects with benign lung nodules were enrolled from a hospital in Dalian, Liaoning Province. We selected 47 serum amino acid and carnitine indicators from targeted metabolomics studies using LC‒MS/MS and age and sex demographic indicators of the subjects. After screening by a stepwise regression algorithm, 16 metrics were included. The XGBoost model in the machine learning algorithm showed superior predictive power (AUC = 0.81, accuracy = 75.29%, sensitivity = 74%), with the metabolic biomarkers ornithine and palmitoylcarnitine being potential biomarkers to screen for lung cancer. The machine learning model XGBoost is proposed as an tool for early lung cancer prediction. This study provides strong support for the feasibility of blood-based screening for metabolites and provide a safer, faster and more accurate tool for early diagnosis of lung cancer.ConclusionsThis study proposes an interdisciplinary approach combining metabolomics with a machine learning model (XGBoost) to predict early the occurrence of lung cancer. The metabolic biomarkers ornithine and palmitoylcarnitine showed significant power for early lung cancer diagnosis.

A Deep Learning Approach to Analyze NMR Spectra of SH-SY5Y Cells for Alzheimer’s Disease Diagnosis

The SH-SY5Y neuroblastoma cell line is often used as an in vitro model of neuronal function and is widely applied to study the molecular events leading to Alzheimer’s disease (AD). Indeed, recently, basic research on SH-SY5Y cells has provided interesting insights for the discovery of new drugs and biomarkers for improved AD treatment and diagnosis. At the same time, untargeted NMR metabolomics is widely applied to metabolic profile analysis and screening for differential metabolites, to discover new biomarkers. In this paper, a compression technique based on convolutional autoencoders is proposed, which can perform a high dimensionality reduction in the spectral signal (up to more than 300 times), maintaining informative features (guaranteed by a reconstruction error always smaller than 5%). Moreover, before compression, an ad hoc preprocessing method was devised to remedy the scarcity of available data. The compressed spectral data were then used to train some SVM classifiers to distinguish diseased from healthy cells, achieving an accuracy close to 78%, a significantly better performance with respect to using standard PCA-compressed data.

Pharmacokinetics and metabolomics of the new psychoactive substance 4-chloroethylcathinone

Synthetic 4-Chloroethcathinone (4-CEC) is a derivative of cathinone that belongs to one of the more severe abused substances among new psychoactive substances (NPS). Current researches on 4-CEC mainly focus on metabolite identification studies, and there is a lack of researches on pharmacokinetic, tissue distribution and metabolomics studies in vivo. A sensitive and reliable LC-MS/MS assay was developed and validated for the determination of 4-CEC concentrations in plasma and tissue homogenates. According to the pharmacokinetic results, the absorption and elimination of 4-CEC were faster after administration. The Cmax was 1896 ± 876 ng/ml, the peak time Tmax was 10.1 ± 9.2 min, and the elimination half-life t1/2 was 100.4 min. Metabolomics studies showed that the highest concentrations of 4-CEC were found in brain, lung, kidney and liver. The results of tissue biopsy showed that the liver, kidney and brain tissue had a certain degree of damage. After 4-CEC administration, amino acid-related metabolism and biosynthesis, lipid metabolism, niacin and niacinamide metabolism in mice were interfered, suggesting that 4-CEC could cause energy metabolism disorder in mice. The metabolic pathways and toxicity mechanisms related to 4-CEC entry into the body were explained at the overall metabolic level by multivariate data analysis, screening and identification of differential metabolites and metabolic pathway analysis.

Clinical Presentation of Inherited Metabolic Diseases in Newborns Hospitalised in an Intensive Care Unit.

The first clinical manifestations of inherited metabolic diseases occur in the neonatal period in up to half of cases, often with nonspecific symptoms, making their recognition challenging. This study aimed to characterise inherited metabolic disease cases with neonatal presentation requiring admission to the paediatric intensive care unit in a Portuguese reference centre for inherited metabolic diseases. An observational study with retrospective data collection was performed, including all newborns with an inherited metabolic disease admitted to the pediatric intensive care unit between June 2011 and June 2022. Three 'pathophysiological' groups were defined: cases due to small molecules, energy deficiency and complex molecules. Twenty newborns, with a median age at admission of 7.5 days, were included. Thirteen (65%) were female, sixteen (80%) had a small molecule disorder, and four (20%) had diseases of energy defects. Neurological manifestations were the most common, with most newborns presenting symptomatically in the first week of life. There was no difference between the groups in neurological, cardiac, and hepatic involvement and shock at presentation. A symptom-free interval was more frequent in patients with small molecule disorders than the others (p=0.01). The main metabolic changes found were altered plasma amino acids (n=13) and organic aciduria (n=10), creatine kinase elevation (n=13), hyperlactatemia (n=12), metabolic acidosis with increased anion gap (n=8) and hyperammonaemia (n=7). Newborn screening of metabolites helped make a diagnosis in 60% of cases. Five newborns died due to multiorgan failure (n=3) or refractory cardiogenic shock (n=1), and in one, therapeutic efforts were limited due to an adverse neurological prognosis. Although the symptoms and signs are often nonspecific, we should suspect inherited metabolic disease when a newborn presents with neurological symptoms after a symptom-free period, however short it might be. Newborns with suspected inherited metabolic disease should be evaluated with simple biochemical tests, and newborn screening should be urgently expanded to start specific treatment earlier, reducing mortality and morbidity.

Screening of secondary metabolites produced by a mangroove-derived Nigrospora species an endophytic fungus isolated from Rhizophora racemosa for antioxidant and antimicrobial properties

Marine fungi have shown large chemo-diversity of unharnessed important secondary metabolites required for drug development. In this study, endophytic fungi Nigrospora species (5S3) was isolated from the fresh stem of Rhizophora racemosa from a mangrove forest in Lagos Nigeria and axenic colonies were fermented on sterile rice medium for 21 days. The antimicrobial and antioxidant activities of the extract were evaluated. Chemical analyses such as High Performance Liquid Chromatography Diode Array Detector (HPLC-DAD) and Nuclear magnetic Resonance Imaging (NMR) analyses of selected Vacuum Liquid Chromatography (VLC) fractions revealed the presence of ten known compounds with established biological activities. Tested fractions of the extract exhibited antimicrobial activities against at least one Gram positive and Gram negative bacteria with MIC values that ranged between 0.13 to 1 mg/mL. FE2 and FE3 fractions of the extract demonstrated poor antioxidant activity of less than 10%. Septicine, Aureonitol, Papuamine, Di-iso-octylphtalat, 1-(2,4 Dihydroxy-3,5-Dimethylphenyl)-Ethanone, Cladosporin, Tetrabenzofuran, Dihydrophthalate, 9-Octadecaenoic acid and Eicosane were the compounds detected in the extracts. Our findings reveal that Nigrospora spp an endophytic fungi possesses unique chemo diversity of bioactive secondary metabolites that could be utilized for development of new drugs.

Screening of secondary metabolites produced by a mangroove-derived Nigrospora species an endophytic fungus isolated from Rhizophora racemosa for antioxidant and antimicrobial properties

Screening of secondary metabolites produced by a mangroove-derived Nigrospora species an endophytic fungus isolated from Rhizophora racemosa for antioxidant and antimicrobial properties

High-throughput virtual screening of marine algae metabolites as high-affinity inhibitors of ISKNV major capsid protein: An analysis of in-silico models and DFT calculation to find novel drug molecules for fighting infectious spleen and kidney necrosis virus (ISKNV)

Infectious Spleen and Kidney Necrosis Virus (ISKNV) is linked to severe infections that cause significant financial losses in global aquaculture. ISKNV enters the host cell through its major capsid protein (MCP), and the resulting infection can lead to mass mortality of fish. Even though several drugs and vaccines are at various stages of clinical testing, none are currently available. Thus, we sought to assess the potential of seaweed compounds to block viral entrance by inhibiting the MCP. The Seaweed Metabolite Database (1110 compounds) was assessed for potential antiviral activity against ISKNV using high throughput virtual screening. Forty compounds with docking scores of ≥8.0 kcal/mol were screened further. The inhibitory molecules BC012, BC014, BS032, and RC009 were predicted by the docking and MD techniques to bind the MCP protein significantly with binding affinities of −9.2, −9.2, −9.9, and −9.4 kcal/mol, respectively. Also, ADMET characteristics of the compounds indicated drug-likeness. According to this study, marine seaweed compounds may operate as viral entrance inhibitors. For their efficacy to be established, in-vitro and in-vivo testing is required.

Transcriptome and Metabolome Integration Reveals the Impact of Fungal Elicitors on Triterpene Accumulation in Sanghuangporus sanghuang.

Sanghuangporus sanghuang is a large wood-decaying mushroom highly valued in traditional Chinese medicine due to its medicinal properties, including hypoglycemic, antioxidant, antitumor, and antibacterial properties effects. Its key bioactive compounds include flavonoids and triterpenoids. Specific fungal genes can be selectively induced by fungal elicitors. To investigate the effect of fungal polysaccharides derived from Perenniporia tenuis mycelia on the metabolites of S. sanghuang, we conducted metabolic and transcriptional profiling with and without elicitor treatment (ET and WET, respectively). Correlation analysis showed significant differences in triterpenoid biosynthesis between the ET and WET groups. In addition, the structural genes associated with triterpenoids and their metabolites in both groups were verified using quantitative real-time polymerase chain reaction (qRT-PCR) and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Through metabolite screening, three triterpenoids were identified: betulinol, betulinic acid, and 2-hydroxyoleanolic acid. Excitation treatment increased the level of betulinic acid by 2.62-fold and 2-hydroxyoleanolic acid by 114.67-fold compared to WET. The qRT-PCR results of the four genes expressed in secondary metabolic pathways, defense gene activation, and signal transduction showed significant variation between the ET and WET groups. Overall, our study suggests that the fungal elicitor induced the aggregation of pentacyclic triterpenoid secondary metabolites in S. sanghuang.

Biological significance of carbohydrate active enzymes and searching their inhibitors for therapeutic applications

Glycans are the most abundant and diverse group of biomolecules with a crucial role in all the biological processes. Their structural and functional diversity is not genetically encoded, but depends on Carbohydrate Active Enzymes (CAZymes) which carry out all catalytic activities in terms of synthesis, modification, and degradation. CAZymes comprise large families of enzymes with specific functions and are widely used for various commercial applications ranging from biofuel production to textile and food industries with impact on biorefineries. To understand the structure and functional mechanism of these CAZymes for their modification for industrial use, together with knowledge of therapeutic aspects of their dysfunction associated with various diseases, CAZyme inhibitors can be used as a valuable tool. In search for new inhibitors, the screening of various secondary metabolites using high-throughput techniques and rational design techniques have been explored. The inhibitors can thus help tune CAZymes and are emerging as a potential research interest.

GPAM mediated lysophosphatidic acid synthesis regulates mitochondrial dynamics in acute myeloid leukemia.

Metabolic alterations, especially in the mitochondria, play important roles in several kinds of cancers, including acute myeloid leukemia (AML). However, AML-specific molecular mechanisms that regulate mitochondrial dynamics remain elusive. Through the metabolite screening comparing CD34+ AML cells and healthy hematopoietic stem/progenitor cells, we identified enhanced lysophosphatidic acid (LPA) synthesis activity in AML. LPA is synthesized from glycerol-3-phosphate by glycerol-3-phosphate acyltransferases (GPATs), rate-limiting enzymes of the LPA synthesis pathway. Among the four isozymes of GPATs, glycerol-3-phosphate acyltransferases, mitochondrial (GPAM) was highly expressed in AML cells, and the inhibition of LPA synthesis by silencing GPAM or FSG67 (a GPAM-inhibitor) significantly impaired AML propagation through the induction of mitochondrial fission, resulting in the suppression of oxidative phosphorylation and the elevation of reactive oxygen species. Notably, inhibition of this metabolic synthesis pathway by FSG67 administration did not affect normal human hematopoiesis in vivo. Therefore, the GPAM-mediated LPA synthesis pathway from G3P represents a critical metabolic mechanism that specifically regulates mitochondrial dynamics in human AML, and GPAM is a promising potential therapeutic target.

Untargeted screening of plant metabolites based on data-independent and data-dependent acquisition modes using LC-ESI-QTOF-MS: Tribulus terrestris L. as a case study

Metabolomics has been used as a powerful tool for the analysis, and drug-lead identification in medicinal plants and herbal medicines. For the coverage of a broader range of plant-based metabolites using LC-MS, one of the important parameters is the selection of analysis mode and data processing for identification. This paper describes the utility of two distinctive acquisition modes in combination, a classic data-dependent acquisition (DDA) mode and a data-independent acquisition (DIA) mode for obtaining the mass spectrometry data of plant extracts using LC-ESI-QTOF/MS. Tribulus terrestris plant was used as a case study. We have applied three-step data analysis pipeline 1-annotation and putative identification of metabolites, 2-validation, and relative quantification, and 3-multivariate analysis using open-access MS-DIAL, Skyline, and Perseus software. A total of four samples of T. terrestris (aqueous extracts), two fruits, and two whole plant samples, from two different regions, were used. By combining data analysis results of plant fruit samples from two different regions, a total of 95 and 77 metabolites were identified in positive and negative ionization modes, respectively. Similarly, in the analysis of the whole plant from two different regions, 75 and 76 metabolites were identified in positive and negative ionization modes, respectively. We suggested the use of DDA mode for annotation, identification of metabolites, and generation of transition lists in MS-DIAL, furthermore, the use of DIA acquisition mode for enhancing metabolites sensitivity in complex samples, deconvolute MS1/MS2 spectra in Skyline for the quantitative performance and analytical reliability. The developed protocol can be used for the broader coverage of plant-based metabolites.

Hepatoprotective effect of the Hexane extract of Luffa echinata root and Stem bark against Tetrachloromethane Induced Hepatic Disorder in rats

In many nations, long term liver syndromes are the principal cause of casualty. Medicinal drug obtained from fauna and floras are well-thought-out and valuable in the treatment of chronic diseases that require long-term treatment. An experiment was conducted to determine the potential outcome of a hexane extract of the root and stem bark of Luffa echinata (L.echinata) Roxb on tetra chloromethane (TCM) induced liver toxicity in female albino rats. The root and stem bark of L. echinata were treated with hexane (C6H6), and exposed to preliminary secondary metabolite screening for the presence of various therapeutic component. Tetra chloromethane has been used to cause the hepatic disease in albino rats, which was subsequently treated with Hexane extract. Biochemical markers including entire bilirubin (BR), serum glutamic–oxaloacetic transaminase (SGOT) enzyme, serum glutamic –pyruvic transaminase (SGPT) enzyme, and total serum protein (TSP) in blood serum were used to assess the patient. The synergistic outcome of root and stem bark hexane extract on the damage liver tissues were also investigated histopathologically. According to the findings, Hexane extract has a strong hepatoprotective effect.

A mass defect filtering combined background subtraction strategy for rapid screening and identification of metabolites in rat plasma after oral administration of Yindan Xinnaotong soft capsule

The absorbed prototypes and metabolites of traditional Chinese medicines (TCMs) serves an important part in pharmacological action and clinical effects. However, the comprehensive characterization of which is facing actual or possible rigorous challenges due to the lack of data mining methods and the complexity of metabolite samples. Yindan Xinnaotong soft capsule (YDXNT), a typical traditional Chinese medicine prescription consisting of extracts from 8 herbal medicines, is widely used for the treatment of angina pectoris and ischemic stroke in the clinic. This study established a systematic data mining strategy based on ultra-high performance liquid chromatography tandem quadrupole-time-of-fight mass spectrometry (UHPLC-Q-TOF MS) for comprehensive metabolite profiling of YDXNT in rat plasma after oral administration. The multi-level feature ion filtration strategy was primarily conducted through the full scan MS data of plasma samples. All potential metabolites were rapidly fileted out from the endogenous background interference based on the background subtract and the chemical type specifically mass defect filter (MDF) windows including flavonoids, ginkgolides, phenolic acids, saponins, and tanshinones. As the MDF windows of certain types were overlapped, the screened-out potential metabolites were deeply characterized and identified according to their retention times (RT), integrating neutral loss filtering (NLF), diagnostic fragment ions filtering (DFIF), and further confirmed by reference standards. Thus, a total of 122 compounds, consisting of 29 prototype components (16 confirmed with reference standards) and 93 metabolites had been identified. This study provides a rapid and robust metabolite profiling method for researching complicated traditional Chinese medicine prescriptions.

A Preliminary Phytochemical, Pharmacognostic and Physicochemical Evaluation of Endemic Thymus spathulifolius (Lamiaceae)

Aim of study: The members of the genus Thymus are widely used in Turkey as herbal teas, spices and salads for different purpose and the herbs of Thymus spathulifolius are reported to have medicinal value. Therefore, in this study various pharmacognostic parameters, physicochemical properties and anatomical characters of Thymus spathulifolius Hausskn. & Velen., is a medicinal plant and endemic to Turkey, were investigated. Area of study: T. spathulifolius was dried and stored according to herbarium techniques in the Herbarium of Pharmacy Faculty, Trakya University, collected from Sivas, Turkey. Material and methods: The total ash, moisture content, and physicochemical tests on the different extracts obtained from the endemic T. spathulifolius aerial parts collected from Sivas province were determined according to the World Health Organization (WHO) standards for crude drugs. Main results: The moisture content and total ash amount of the drug obtained from this plant is in accordance with the WHO standards. Phytochemical screening for secondary metabolites showed the presence of carbohydrates, antraquinone glycosides, steroids, flavonoids and coumarins, while other phytochemicals such as alkaloids and saponins were absent. Besides, the cross-sections of root, stem and leaf were examined and detailed. The total ash value was found as 7.36 ± 0.21% and acid insoluble ash was 0.14 ± 0.72%. Highlights: The present study provided important information for correct authentication and standardization of this plant material.

A multi-aspect analysis of two analogous aspergillus spp. belonging to section Flavi: aspergillus flavus and aspergillus oryzae

Microfungal isolates were routinely identified depending on both macro and micro morphological characteristics, sometimes, some fungal isolates appeared to be similar and such cases caused severe confusion for mycologists during the preliminary identification. During our previous studies dealing with isolation of fungi for some biotechnological applications; two mystifying species Aspergillus flavus and Aspergillus oryzae showed similar cultural and macroscopic features. Therefore, the current study aimed to easily distinguish between these two species depending on simple approaches which are routinely followed by a large segment of researchers. Investigation of the macroscopic features was performed to check the fungal growth on four different media (PDA, MEA, YES, and CYA) followed by microscopic examination using an ordinary light microscope, and scanning electron microscope SEM. Also, screening of secondary metabolites for both strains was preliminarily identified to find out the difference between their metabolic profiles. Finally, ITS rDNA was involved to clarify the molecular differences along their partial sequence. Conclusively, the BLAST strategy confirmed the similarity of ITS rDNA segments of both fungal strains that supported our hypothesis. The color of the fungal growth is a very critical factor whereas it is extensively influenced by the type of cultivation media. Accordingly, the YES medium was an inspiring tool assisting in prompt differentiation during the culture investigation step whereas A. oryzae and A. flavus appeared significant mustard yellow and olive green respectively. During the microscopic examination, the CYA medium also had a robust effect on the formation of the conidial chain whereas the knit long chain was observed in A. oryzae while the conidia appeared scattered and not in a chain in the case of A. flavus. Likewise, both two strains possessed different metabolic profiles where A. oryzae is not an Afla toxin producer, unlike A. flavus.