Josef Hyánek*, František Pehal, Kseniya Dryahina, Ladislava Dubská, Blanka Míková, Lada Gombíková, Miroslav Průcha, Stanislav Kubů, Petra Haláčková and Jaroslav Feyreisl
Introduction: In this retrospective study, we comment on the cause and diagnostic potential of the elevated serum total cholesterol and some non-cholesterol sterols in a population of healthy pregnant women from Prague, Czech Republic.
Methods: Based on a total of 21,000 clinical biochemistry tests of healthy pregnant women with hypercholesterolemia observed during pregnancy, a testing group of 84 women with a total cholesterol (TC) above 7.0 mmol/l was established to analyze their non-cholesterol sterols (NCS) by Gas Chromatography–Mass Spectrometry. Lathosterol (Lat) and desmosterol (Des) were evaluated as markers of endogenous cholesterol synthesis, whereas campesterol (Cam) and sitosterol (Sit) were analysed as markers of intestinal absorption.
Results: In the basic population, the frequency of gestational hypercholesterolemia with the serum TC levels > 7.0mmol/l was 1 to 136.The mean values were: TC 6.8 mmol/l, LDL-C 4.6 mmol/l, and HDL-C 2.2 mmol/l. In the selected testing group of 84, the mean values were: Lat 7.8+/-1.7 μmol/l, Des 4.7+/-0.9 μmol/l, Cam 9.8+/-2.6 μmol/l, and Sit 9.6+/-3.8 μmol/l. Lat correlated with TC (r = 0.53), LDL-C (r = 0.36), and non-HDL-C (r = 0.35). No such correlations were observed for Cam or Sit.
Conclusion: Our findings prove that gestational hypercholesterolemia is caused by increased endogenous cholesterol synthesis via lathosterol. Subsequently, we demonstrate how a single cholesterol test taken in the fifth to sixth month gestation can efficiently help detect familial hypercholesterolemia, and prevent related late pregnancy circulatory complications.
Background: The proposal that MALDI-ToF mass spectrometry could be used as a direct, rapid and affordable diagnostic tool in clinical laboratory medicine has moved from a theoretical possibility to a reality for Microbiology. Several studies have proposed the application of this technology in obstetric and gynaecological evaluation of patients. In particular, we have proposed that the adoption of MALDI-ToF mass spectrometry in examination of maternal pregnancy urine samples for the detection of Downs syndrome.
Methods: A retrospective collection of 20 Down Syndrome and 100 non-aneuploid pregnancy urines at 12 to 14 weeks gestation, collected in 2007-2008 from high risk pregnancy cohorts, were examined by MALDI-ToF mass spectrometry in the mass/charge range between 1000 and 100000 m/z. Normalisation of spectral data was defined using mass bins of 100 m/z expressed as a percentage of the total ion count of the mass spectra from 2000 to 11000 m/z. Of the ninety 100 m/z bins, forty-six were identified as m/z bins at which statistically significant differences in spectra occurred between Downs and control/non-aneuploid samples. Based on the differences and variance, for values at these bins, weighted scores of the probability of being Downs were assigned. Comparative algorithms consisting of various mass bins were tested for ability to distinguish Down syndrome from non-aneuploid pregnancy.
Results: Although various algorithms could distinguish Downs from non-aneuploid controls, it was found that gestational age was a confounding factor and that if separated into gestational age matched cohorts the ability to distinguish the groups improved dramatically e.g. whilst a 19 bins algorithm separated 100% of Downs from non-aneuploid pregnancies for a 9% false positive rate in the mixed gestational ages group; a two bin algorithm distinguished 100% of Downs for a 6% false positive rate for the 12 weeks gestational age pregnancies.
Conclusion: Normalised MALDI-ToF mass spectra, at 2000 to 11000 m/z, of maternal urine gives rise to gestational age specific screening tests algorithms for Downs’s syndrome.
Jason W Birkett*, Ellen Goudsmits and George P Sharples
Published on: 5th September, 2017
The application of monolithic material sorption extraction, specifically MonoTrapTM, to the extraction of organic gunshot residue (OGSR) compounds from unburnt propellant powders is described. Four different MonoTraps were assessed for their capability to extract OGSR compounds from two different ammunition types. Extracts were analysed using gas chromatography-mass spectrometry. Results indicated that the carbon disc was suitable for the extraction of OGSR compounds from unburnt propellant powders. Quantities for major compounds were comparable to methanol extractions. The method was successfully used to detect a wide range of OGSR compounds, including nitrotoluenes, nitroglycerin, diphenylamines and centralites and is expected to be applicable to a wide range of ammunition types.
Background: The plant Artemisia annua has been used in traditional Chinese medicine for many years. Rich in bioactive molecules, the A. annua plant is used to extract the anti-malaria compound artemisinin (< 1%), which results in most of the plant being unutilized. One byproduct of artemisinin extraction is artemisia naphtha (AN), which has yet to be studied extensively.
Aims: Study the activity of a novel AN oil extract against microbes, pro-inflammatory cytokines, and dermatological endpoints that are key for eczema and acne pathogenesis to determine if an effective A. annua extract for these skin conditions can be developed.
Methods: Gas chromatography-mass spectrometry was performed to determine the composition of AN oil. P. acnes, S. aureus, M. furfur, and C. albicans were cultured to determine minimal inhibitory concentration. in vitro studies utilizing keratinocytes and macrophages were treated with AN oil and gene expression measured by quantitative RT-PCR. A 13-subject clinical trial was performed with 1% AN oil Gel to assess its potential benefits for sensitive and acne prone skin.
Results: AN oil upregulates filaggrin gene expression and possesses antimicrobial and anti-inflammatory activity inhibiting LPS, S. aureus and "Th2 induced" pro-inflammatory mediator release (IL-6, IL-8 and TSLP). Clinical assessment of 1% AN Gel shows it reduces acne blemishes and the appearance of redness.
Conclusion: Previously an underutilized and unpurified byproduct, AN is now the source to develop the first topical AN oil for cosmetic use with an activity profile that suggests it is effective for those with sensitive and/or acne prone skin.
In this study we describe a method for the detection of biomolecules (in the polypeptide m/z range) directly from the surface of plant leaves by using Mass Spectrometry Imaging. The plant-pathogen interaction between Arabidopsis thaliana and the bacterium Xanthomonas campestris pv. campestris was analyzed by comparing infected and non-infected leaf discs submitted to mass spectrometry. The total surface area of ion distribution was calculated for both samples, revealing 23 ions, out of which 3 showed statistical significance. Although these ions were not identified, the results showed that this approach can be successfully applied for the detection of potential polypeptide biomarkers directly on leaf tissue, which is a major challenge in MALDI-Imaging studies.
Laura Silva Fernandes, Ygor Ferreira Garcia da Costa, Martha Eunice de Bessa, Adriana Lucia Pires Ferreira, José Otávio do Amaral Corrêa, Glauciemar Del-Vechio Vieira, Orlando Vieira de Sousa, Ana Lúcia Santos de Matos Araújo, Paula C Castilho* and Maria Silvana Alves*
Morbidity and mortality of the infected patients by multidrug-resistant bacteria have increased, emphasizing the urgency of fight for the discovery of new innovative antibiotics. In this sense, natural products emerge as valuable sources of bioactive compounds. Among the biodiversity, Eryngium pristis Cham. & Schltdl. (Apiaceae Lindl.) is traditionally used to treat thrush and ulcers of throat and mouth, as diuretic and emmenagogue, but scarcely known as an antimicrobial agent. With this context in mind, the goals of this study were to investigate the metabolic profile and the antibacterial activity of ethanolic extract (EE-Ep) and hexane (HF-Ep), dichloromethane (DF-Ep), ethyl acetate (EAF-Ep) and butanol (BF-Ep) fractions from E. pristis leaves. Gas Chromatography-Mass Spectrometry (GC-MS) was performed to stablish the metabolic profile and revealed the presence of 12 and 14 compounds in EAF-Ep and HF-Ep, respectively. β-selinene, spathulenol, globulol, 2-methoxy-4-vinylphenol, α-amyrin, β-amyrin, and lupeol derivative were some of phytochemicals identified. The antibacterial activity was determined by Minimal Inhibitory Concentration (MIC) using the broth micro-dilution against eight ATCC® and five methicillin-resistant Staphylococcus aureus (MRSA) clinical strains. HF-Ep was the most effective (MIC ≤ 5,000 µg/µL), being active against the largest part of tested Gram-positive and Gram-negative bacterial strains, including MRSA, with exception of Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 9027) and (ATCC 27853). These results suggest that E. pristis is a natural source of bioactive compounds for the search of new antibiotics which can be an interesting therapeutic approach to recover patients mainly infected by MRSA strains.
Bhagat Singh*, Satish R Nailkar, Chetansen A Bhadkambekar, Suneel Prajapati and Sukhminder Kaur
Published on: 28th March, 2023
Prescriptions for psychoactive substances such as Pregabalin, Methamphetamine, Caffeine, Clonazepam and Mirtazapine (PM-CCM) are common in the treatment of a variety of disorders. Indeed, the PM-CCM has been used in different therapeutic areas, including insomnia, anxiety, seizure disorders, etc. Unfortunately, these psychoactive substances are present in the illegal street market, leading to a lot of drug abuse among some addicted users, road insecurity and suicide. Hence, it has become essential to validate and develop a rapid and effective method to analyze the PM-CCM, a modified illicit drug, for drug abuse in the forensic sciences. A simple, rapid, specific and sensitive Gas Chromatography-Mass Spectrometry(GC-MS) method has been developed for the identification of Pregabalin, Methamphetamine, Caffeine, Clonazepam and Mirtazapine (PM-CCM) in forensic exhibits. At room temperature, the sample was ultrasonicated for 5 minutes before being extracted with methanol. A highly precise auto-injector is used to inject a very small quantity of samples for analysis. Helium is used as a carrier gas with a flow rate of 1 ml/min. The separation of PM-CCM was performed on SH-RXi-5 MS, ID.25 mm, film thickness. 25 µm, length of 30 m column. The constituents of PM-CCM were identified by the mass-to-charge ratio (m/z ratio) of fragments of the parent compound by comparing them with the NIST-17 MS Library. Separation and identification of PM-CCM were achieved within a 15-minute run. The proposed method has been successfully used for the routine analysis of PM-CCM in complex illicit drug preparations and in forensic exhibits as well. The application of above discussed qualitative analysis method and screening of PM-CCM, modified illicit drug samples demonstrates the potential and applicability of the technique to the fast chemical profiling of illicit samples.
Randa A Almahdi*, Hiba Abdel Rahman Ali and Sami Ahmed Khalid
Published on: 27th December, 2023
Introduction: In Sudan, Grewia tenax fruits, are known commonly as Goddaim. The fruit’s pericarp is used traditionally for a long time as a juice or a porridge to treat iron deficiency anemia (IDA). Traditional Goddaim users have a very strong belief in its effectiveness. However, the pattern of hemoglobin improvement follows an initial fast increase followed by a decline upon continuing its use. Some previous studies have attributed its effect to high iron content, while the iron quantity was too small. This work attempts to find an explanation for its mechanism of action by screening the fruit extract and its respective fractions for secondary metabolites, minerals, vitamins, and fibre. Methods: Entailed three methodologies: Chemical analysis to identify quantified minerals, ascorbic acid and non-digestible fibers, Phytochemical Analysis to separate and identify secondary metabolites using high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS-MS technique), and estimation of radical scavenging activities of crude fruit extract and its respective chloroform and ethyl acetate fractions by inhibition of the 2,2-diphenylpicrylhydrazyl (DPPH).Results: Ascorbic acid and indigestible fibres were revealed in the aqueous extract. Secondary metabolites were flavonoids (e.g., quercetin, kaempferol), organic acids (e.g., ferulic acid, chlorogenic acid), β-carboline alkaloids (e.g., 3-hydroxy-tetrahydroharman) identified in the chloroform, and ethyl acetate fractions. The in vitro antioxidant activity of G. tenax fruit extract was confirmed spectrophotometrically. Conclusion: It can be concluded that the initial enhancement of iron absorption through ascorbic acid and fibres, followed by iron uptake inhibition, could be explained by iron chelation by the chelators in the fruits. This paradoxical effect may qualify G. tenax fruits as a safety gauge for improving haemoglobin levels without compromising iron excess once iron stores are filled by keeping the oxidative stress in check. This may present G. tenax fruits as a good and safe remedy that optimizes the treatment of IDA.
In order to study the molecular mechanism of the antioxidant effect of enzymatically hydrolyzed tuna dark meat peptides, this article uses alkaline protease to enzymatically hydrolyze tuna dark meat, and at the same time performs peptide sequencing using matrix-assisted laser dissociation time-of-flight mass spectrometry (MALDI TOF/TOF). Discovery Studio (DS) performed molecular docking. Finally, the antioxidant effect was verified through DPPH clearance experiments. The results show that the dominant peptide sequences in the tuna dark meat hydrolyzed polypeptides are LAPGQ, GGGDPI, and PLRLP; through molecular simulation methods (Discover Studio, DS), the potential target of the above-mentioned enzymatic polypeptides was screened out to be Keap1, thus predicting antioxidant activity. It provides theoretical support for further research on enzymatic peptides. Through DPPH clearance experiments, it was found that both the enzymatic hydrolysate and LAPGQ, GGGDPI, and PLRLP have antioxidant activity, confirming their effects.
Noor Jawad Khadhum, Neepal Imtair Al-Garaawi* and Antethar Jabbar Al-Edani
Published on: 25th June, 2024
The current study included a variety of phytochemical substances that were extracted from Centaurium pulchellum and showed a wide range of medicinal properties from the plant's reproductive and vegetative parts against the pathogenic fungus Aspergillus flavus. The vegetative and reproductive components of Centaurium pulchellum were subjected to (GC-MS) analysis for phytochemical study. The data indicated that fungal activity was the highest. Four extract concentrations of 5, 10, 15, and 20 mg/ml were utilized in the investigation, and the diameter of the colonies measured at each concentration was 90.00, 36.00, 28.00, 18.00, and 0.00 mm, respectively.Nine bioactive phytochemical compounds were found in Centaurium pulchellum's vegetative and reproductive portions, according to GC-MS analysis of the chemicals. Another study reported phytochemical substances that: 1-H-Imidazole-2-carboxaldehyde, 1-methyl-;Acetaminophen; n-Hexadecanoic acid; Mercaptoacetic acid, 2TMS derivative; 1.2,3-Dimethyl-5-(trifluoromethyl)-1,4-benzenediol #; Mercaptoethanol, 2TMS derivative-; Bis-(3,5,5-trimethylhexyl) phthalate Tetrakis(trimethylsilyl) orthosilicate #;- 1.1-Isopropoxy-3,3,3-trimethyl-1-[(trimethylsilyl)oxy]disiloxanyl tris(trimethylsilyl) orthosilicate #.
Peguy Flora Djuidje Kouomou*, Esperanza Ghomsi Guiadem, Maurice Fotsing Tagatsing, Wong Mui-Yong, Carole Beaulieu, Cecile Annie Ewane and Thaddee Boudjeko
Published on: 18th November, 2024
Background: Microorganisms belonging to Streptomyces sp. are Gram-positive bacteria known for their unsurpassed capacity for the production of secondary metabolites with diverse biological activities. The aim of this study was to evaluate the antimicrobial and antioxidant properties of ethyl acetate Streptomyces sp. PERM2 extract, its potential modes of action and bioactive secondary metabolites.Results: The ethyl acetate PERM2 extract showed antimicrobial activity more pronounced on both Gram-positive and Gram-negative bacteria and fungi with a Minimum Inhibitory Concentration value (MIC) of 0.5 mg/mL and Minimum Bactericidal Concentration (MBC) of 2 - 4 mg/mL against bacterial pathogens. MIC value against pathogenic fungi was 2 mg/mL and Minimum Fungicidal Concentration (MFC) of 0.01 - 0.05 mg/mL against pathogenic fungi. PERM2 crude extract showed the ability to inhibit bacteria cell wall synthesis at 0.5 and 1 MIC. The extract was found to possess dose-dependent 2,2-Diphenyl-picrylhadrazyl (DPPH) free radical scavenging and Ferric reducing activity. The gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of three major compounds identified as 9,12-octadecadienoic acid (Z, Z) (29.75%), tridecyl trifluoroacetate (24.82%) and 1-(+)-ascorbic acid 2, 6-dihexadecanoate (22.34%). The liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis revealed the presence of 22 non-volatile metabolites in PERM2 extract and only the compound 3, 30-O-dimethylellagic acid was identified. Conclusion: The results of this study indicate that ethyl acetate Streptomyces sp. PERM2 extract possesses antibacterial, antifungal, and antioxidant activities; inhibits bacteria cell wall and protein synthesis; and contains significant bioactive secondary metabolites which could be used as an alternative to multi-resistance antibiotics.
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