Shimadzu GC-2010

Background. In developing countries, the prevalence of bacterial infections is quite rampant due to several factors such as the HIV/AIDS pandemic, lack of hygiene, overcrowding, and resistance to conventional antimicrobials. Hence the use of plant-based antimicrobial agents could provide a low-cost alternative therapy. Rosmarinus officinalis is reputed as a medicinal plant in Ethiopia; however, its antibacterial activity against many of the clinical isolates remains overlooked. Methods. Tender foliage of R. officinalis was collected and extracted in ethanol (EtOH) and evaluated for their antimicrobial activity against ten multidrug-resistant (MDR) clinical isolates, human type culture pathogens, and meat-borne bacterial isolates by employing agar well diffusion assay. Results. EtOH extract of R. officinalis efficiently subdued the growth of all tested MDR clinical isolates in varying degrees. Salmonella sp. and Staphylococcus aureus were found to be the most sensitive clinical isolates. Likewise, it efficiently repressed the growth of meat-borne pathogens, particularly, S. aureus and Salmonella sp. showing its potentiality to be used as a natural antibacterial agent in the meat processing industry. The mechanism of antibiosis of plant extract against meat-borne pathogens is inferred to be bactericidal. Chemical constituents of the crude plant extract were analysed by Gas Chromatography-Mass Spectroscopy (GC-MS), Fourier Transform Infrared (FT-IR), and UV-visible spectroscopy showing genkwanin (26%), camphor (13%), endo-borneol (13%), alpha-terpineol (12%), and hydroxyhydrocaffeic acid (13%) as the major compounds. Conclusion. Overall results of the present study conclude that R. officinalis could be an excellent source of antimicrobial agents for the management of drug-resistant bacteria as well as meat-borne pathogens.

Aseer Manilal, Kuzhunellil Raghavanpillai Sabu, Melat Woldemariam, Addis Aklilu, Gelila Biresaw, Tsegaye Yohanes, Mohammed Seid, Behailu Merdekios. Evidence-Based Complementary and Alternative Medicine, 2021, 1-10, 2021.

Asarum is a traditional medicine and has been widely used as remedies for inflammatory diseases, toothache, headache, local anesthesia, and aphthous stomatitis in China, Japan, and Korea. Our previous research found that safrole and methyl eugenol were vital compounds that contribute to distinguish the different species and raw Asarum and its processed products apart. The pharmacokinetics of safrole and methyl eugenol after oral administration of Asarum extract has not been reported yet. In this study, a rapid and simple gas chromatography–mass spectroscopy (GC–MS) method that has a complete run time of only 4.5 min was developed and validated for the simultaneous determination and pharmacokinetic study of safrole and methyl eugenol in rat plasma after administration of Asarum extracts. The chromatographic separation was realized on a DB-17 column (). And detection was carried out under selected ion monitoring (SIM) mode. Plasma samples were pretreated by -hexane. The pharmacokinetic parameters provided by this study will be beneficial for further developments and clinical applications of Asarum.

Yuqi Fan, Dongyue Yang, Xuhua Huang, Guangzhe Yao, Wei Wang, Mengyuan Gao, Xiaohua Jia, Huizi Ouyang, Yanxu Chang, Jun He. BioMed Research International, 2021, 1-8, 2021.

Compound-specific stable chlorine isotope analysis (CSIA-Cl) is an important method for identifying sources of organochlorine contaminants and helping assess their quantification of transformation processes. However, the present CSIA-Cl is challenged by either redundant conversion pretreatment or complicated mathematical correction. To overcome the mentioned problems, a novel method has been developed for the CSIA-Cl of eight organochlorine pesticides using gas chromatography-negative chemical ionization mass spectrometry (GC-NCI-qMS) in this study. The instrument parameters, acquisition mode, and required injection amounts were optimized in terms of the precision of GC-NCI-qMS. An ionization energy of 90 eV and emission current of 90 μA were selected, and the precisions for eight organochlorine pesticides were in the range of 0.37‰–2.15‰ in single ion monitoring (SIM) mode when the injected amount was 0.50 mg L−1 (viz. 0.5 ng on column). Furthermore, when standards from Supelco and O2si were calibrated using standards from AccuStandard regarded as external isotope standard, chlorine isotope composition of α-hexachlorocyclohexane (α-HCH) and 2, 2-dichloro−1, 1-bis (4-chlorophenyl) ethylene (p, p′-DDE) in Supelco and O2si was confidently differentiated. The provenance identification method was validated by three organochlorine contaminated groundwater samples and showed a prospect in identifying the source of organochlorine pesticides.

Jing Zhang, Shenghua Liu, Jianye Gui, Xiaoya Li, Guochen Qi. Journal of Analytical Methods in Chemistry, 2021, 1-10, 2021.

To identify a rice genotype with a wider or specific adaptability, the magnitude and patterns of genotype interaction with the environment are valuable clues. However, such information on the nutraceutical lipid compound in rice is limited. In this study, concentrations of five pigmented rice genotypes with different lipid compound levels were evaluated for α-tocopherol, γ-oryzanol, phytosterols, octacosanol, and squalene contents. The research was conducted in eight environments (four locations and two years). Combined analysis of variance showed that levels of α-tocopherol, γ-oryzanol, octacosanol, and squalene were mainly affected by genotype, accounting for more than 42.57% of the total variance. This suggests that improvement of these traits may be achieved by using conventional rice breeding methods. A large proportion of the variation in campesterol and β-sitosterol contents was contributed by the environment. The effect of genotype by environment was significant for all observed traits, especially for γ-oryzanol, β-sitosterol, and squalene. This complicates identification of superior genotypes for these compounds across all environments. Stability parameters showed rice genotypes with high levels of nutraceutical lipid compounds to be sensitive to changes in environmental conditions; thus, specific adaptation is suitable. Our findings suggest that rice breeders and producers should be testing and selecting rice genotypes possessing high added-value lipid concentrations in specific growing conditions or regions.

Bhornchai Harakotr, Kasidid Prompoh, Khundej Suriharn, Kamol Lertrat. International Journal of Agronomy, 2021, 1-10, 2021.

The contents of main components in 45 walnut kernels from 5 walnut-planting provinces in China (Yunnan, Shaanxi, Shandong, Hebei, and Sichuan) were determined using colorimetry, high-performance liquid chromatography, and gas chromatography, including flavonoids, vitamin E, trace elements, fatty acids, and amino acids. The levels of flavonoids and vitamin E were higher in walnuts from Yunnan than in walnuts from the other four provinces. The levels of zinc and iron were relatively higher in walnuts from Shandong, and the calcium content in walnuts from Yunnan was much lower. No obvious difference was found in crude fat concentration for nuts from the five provinces, but differences were observed in the crude protein content. Oleic acid was the predominant unsaturated fatty acids in all walnut species; the monounsaturated fatty acid content was the highest in walnuts from Yunnan. The data obtained here provided insight into differences in walnuts resulting from different growing environments and germplasm genetic traits.

Shuxiang Geng, Delu Ning, Ting Ma, Haiyun Chen, Yinzhi Zhang, Xiulan Sun. Journal of Food Quality, 2021, 1-11, 2021.

The performance characteristics of the engine and the emission levels with Copra Oil (CPO), Virgin Coconut (cocos nucifera) Oil (VCO), Tamanu (calaphyllum inopyllum) Oil (TMO), and Nangae (canarium indicum) Oil (NGO) are presented. The oils, obtained from naturally grown trees in Vanuatu, were tested as straight vegetable oils (SVOs) in a Diesel engine and the results are compared with those of neat diesel. The oils were converted to their fatty-acid-methyl-esters (FAMEs) using gas chromatography to determine their fatty acid compositions. The brake thermal efficiency with SVOs was found to be comparable to diesel. The structure of the alkyl chain and the carbon-to-hydrogen ratio were also studied. All the oils have Palmitic acid, Capric acid, Caprylic acid, and Oleic acid as the major fatty acids. The CPO and VCO have higher amounts of Oleic acid, which acts as an additive and breaks up the interaction between the major fatty acids at higher temperatures, reducing the viscosity. Emissions of CO₂ were lower while those of CO, NOx, and SO₂ were higher with SVOs compared to diesel. The results indicate that the local SVOs are good and inexpensive substitute fuels for Vanuatu that can help the country meet the UN’s sustainable development goals.

Misel J Sisi, M Rafiuddin Ahmed, David Rohindra. Advances in Mechanical Engineering, 12 (9), 168781402096235, 2020.

Background Central nervous system (CNS) infectious diseases are common diseases in emergency rooms and neurology departments. CNS pathogen identification methods are time-consuming and expensive and have low sensitivity and poor specificity. Some studies have shown that bacteria and viruses can produce specific volatile organic compounds (VOCs). The aim of this study is to find potential biomarkers by VOC analysis of cerebrospinal fluid (CSF) in patients with bacterial and viral meningitis/encephalitis (ME). Methods CSF samples from 16 patients with bacterial ME and 42 patients with viral ME were collected, and solid-phase microextraction (SPME) combined with gas chromatography-mass spectrometry (GC-MS) was used to analyze the metabolites in the CSF. Results There are two substances (Ethylene oxide and Phenol) that were found to be different between the two groups. Ethylene oxide was significantly greater in the group of bacterial ME patients than in the viral ME group of patients (P < 0.05). In addition, phenol was remarkably increased in the group of ME patients compared with the bacterial ME patients (P < 0.05). Conclusions Ethylene oxide and phenol may be potential biomarkers to distinguish bacterial ME and viral ME. VOC analysis of CSF may be used as a supporting tool for clinical diagnosis.

Lei Guo, Zhongzhi Qiu, Yue Wang, Kaili Yu, Xiaoya Zheng, Yuhang Li, Miao Liu, Guiyue Wang, Nana Guo, Mengyuan Yang, Enyou Li, Changsong Wang. 2019.

Background: Combined methylmalonic acidemia and homocystinuria, cobalamin C type (cblC defect) is the most common inborn error of cobalamin metabolism. Even with rapid diagnosis and treatment, the long-term outcome remains poor. A reliable method for the prenatal diagnosis of cblC defect is needed for parental decisions regarding continuation of pregnancies of cblC defect fetuses.Results: The between-day and within-day imprecision of Hcy assay were 1.60%∼5.87% and 1.11%∼4.31%, respectively. For the 248 at-risk fetuses, there were 63 affected fetuses with cblC defect and 185 unaffected fetuses. The levels of Hcy in 63 affected fetuses were significantly higher than those in 185 unaffected fetuses, without overlap between the affected and unaffected groups. Sixteen fetuses displayed inconclusive genetic results of MMACHC variants, in which seven fetuses were determined to be affected with elevated amniotic fluid levels of Hcy, C3, C3/C2 and MMA. The remaining nine fetuses were considered unaffected based on a normal amniotic fluid metabolite profile. The diagnostic sensitivities of Hcy and other characteristic metabolites including propionylcarnitine (C3), ratio of C3 to acetylcarnitine (C2; C3/C2), methylmalonic acid (MMA), and methylcitrate acid (MCA) in amniotic fluid were 100%, 87.50%, 100%, 85.71%, and 28.57%, respectively. The respective specificities were 92.05%, 100%, 100%, 97.73%, and 99.43%. Conclusions: Hcy appears to be another sensitive characteristic metabolite biomarker for the prenatal diagnosis of cblC defect. The combination of Hcy assay with acylcarnitine and organic acid analysis is a fast, sensitive, and reliable prenatal diagnostic biochemical approach. This approach could overcome the challenge of the lack of genetic analysis for families with at-risk cblC defect fetuses.

Ting Chen, Lili Liang, Huiwen Zhang, Jun Ye, Wenjuan Qiu, Bing Xiao, Hong Zhu, Lei Wang, Feng Xu, Zhuwen Gong, Xuefan Gu, Lianshu Han. 2020.

Acacia nilotica is an important medicinal plant, found in Africa, Middle East, and Indian subcontinent. Every part of the plant possesses a wide array of biologically-active and therapeutically important compounds and have been used in traditional-system of medicine. We reported the antileishmanial activity of Acacia nilotica (A. nilotica) bark methanolic extract through in vitro assays and dissected the mechanism of its action through in silico studies. Bark methanolic extract exhibited anti-promastigote and anti-amastigote potency with IC50 value of 19.6 + 0.9037 µg/ml and 77.52 + 5.167 µg/ml respectively in time and dose dependent manner. It showed very low cytotoxicity having CC50 value of 432.7 + 7.71 µg/ml on human-macrophage cell line, THP-1. The major constituents identified by GC-MS analysis are 13-docosenoic acid (34.06%), lupeol(20.15 %), 9,12-octadecadienoic acid (9.92 %) and 6-octadecanoic acid (8.43 %) bind effectively with the potential drug-targets of Leishmania donovani (L. donovani ) including sterol 24-c-methytransferase (SMT), trypanothione reductase (TR), pteridine reductase (PTR1) and adenine phosphorybosyl transferase (APRT); suggest the possible mechanism of its antileishmanial action. The highest affinity with all these targets was shown by lupeol. The pharmacokinetic studies, predicted bioactivity scores and acute toxicity studies of major extract constituents support safe antileishmanial drug candidate. This study proved the antileishmanial potential of bark-methanolic extract A. nilotica and its mechanism of action through the inhibition of potential drug targets of L. donovani.

Shams Tabrez, Fazlur Rahman, Bader Alshehri, Abdulaziz Alouffi, Rahat Ali, Fahdah Alshammari, Mohammed Alaidarous, Saeed Banawas, Abdul Aziz Dukhyil, Abdur Rub. 2020.

Background Natural mint flavor is produced by extraction from mint, which is not efficient enough to make it environment friendly process. (−)-Carvone is the monoterpenoid with key flavor of spearmint, and there has been an attempt to produce (−)-carvone by recombinant Escherichia coli. Although all enzymes in (−)-carvone biosynthesis have been functionally expressed in E. coli independently, the yield of (−)-carvone was low in the previous study.ResultsWe have found a by-product formation when cytochrome P450 limonene-6-hydroxylase (P450)/cytochrome P450 reductase (CPR) and carveol dehydrogenase (CDH) were expressed in single cell. These by-products were determined as dihydrocarveol and dihydrocarvone. We hypothesized that the enzymatic kinetics and the expression levels of P450 and CDH are quite different in E. coli. Therefore, two strains independently expressing P450/CPR and CDH were mixed with different mixing ratio, confirming increase in carvone production and decrease in by-product formation when CDH input was reduced. To determine the optimum balance of enzyme expressions, proteome analysis quantification concatemer (QconCAT) method to quantify P450, CPR, and CDH was developed. Using the QconCAT standard protein that was artificially created by concatenating the tryptic peptides, the ratio between P450 and CDH was calculated, and their optimum ratio to maximize (−)-carvone production was shown. Then, a single strain expressing both P450/CPR and CDH was constructed to imitate the superior expression ratio. The upgraded strain showed 15-fold improvement compared to the initial strain, showing 44 ± 6.3 mg/L of (−)-carvone production from 100 mg/L (−)-limonene as a starting substrate.ConclusionsImproved expression balance of P450 and CDH in recombinant E. coli increased the (−)-carvone production using (−)-limonene as the direct substrates by the whole-cell biocatalysis, showing approximately 150 times higher titer than previous report. Our study showed the usefulness of proteome analysis QconCAT method in the strain development for industrial biotechnology field.

Erika Yoshida, Motoki Kojima, Munenori Suzuki, Fumio Matsuda, Akiko Onuki, Yousuke Nishio, Yoshihiro Usuda, Akihiko Kondo, Jun Ishii. 2021.

In this study, Response surface methodology (RSM) was applied to optimize the yield of crude methanolic extract of Mitragyna speciosa leaves using Ultrasound-assisted extraction (UAE). The crude methanolic extract and its fractions were quantified in terms of total phenolic content and total flavonoid content, along with characterized using Fourier-transform infrared and Gas chromatography–mass spectrometry. The results showed the maximum yield of 49.72% at the optimal conditions (temperature, 34 °C; time, 25 min; and volume of solvent, 166 mL). The recovery crude methanolic extract for TPC and TFC were 137.3 ± 15.7 mg GAE/g and 90.3 ± 15.3 mg RE/g, respectively.

Nur Haziqah Zakirat Abd Razak, fazila binti zakaria, Mohd Basyaruddin bin Abdul Rahman, Siti Efliza Ashari. 2020.

Background: Microalgae biofuels have attracted global attention as an alternative to fossil fuels as an energy resource. Microalgae generally accumulate lipids under nitrogen-depleted conditions, but cell growth is depressed under these conditions which causes decrease in lipid productivity. To realize one-step cultivation for biofuel production, microalgae that highly accumulate lipids even under nitrogen-replete conditions are needed. This study aimed to develop a screening method for microalgae mutants with high lipid content even in the presence of a nitrogen source.Results: Mutant cells were generated by irradiating the oleaginous green microalga Chlamydomonas sp. KOR1 with carbon ion beams, cultured under nitrate-replete conditions, and then subjected to FACS-based screening for lipid-rich cells. By repeatedly performing the sequential procedures of cultivation and selection, strains KAC1710 and KAC1801, which highly accumulate lipids under nitrate-replete conditions, were successfully obtained. These mutants formed significant lipid droplets in the cells even in the presence of abundant nitrate and achieved 1.5- and 2.1-fold greater lipid content compared to KOR1, respectively.Conclusion: This study developed a novel nitrogen-conditioned screening method for microalgae mutants that accumulate lipids in the presence of a nitrogen source. This method should contribute to microalgae biofuel production via one-step cultivation under nitrogen-replete conditions.

Tomoki Oyama, Yuichi Kato, Katsuya Satoh, Yutaka Oono, Tomohisa Hasunuma, Akihiko Kondo. 2020.

Osteoporosis is the most common aging-associated bone disease and is caused by hyperactivation of osteoclastic activity. We previously reported that the hexane extract of ginger rhizome [ginger hexane extract (GHE)] could suppress receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells. However, the anti-osteoclastic components in GHE have not yet been identified. In this study, we separated GHE into several fractions using silica gel column chromatography and evaluated their effects on osteoclastogenesis using a RAW264.7 cell osteoclast differentiation assay (in vitro) and the zebrafish scale model of osteoporosis (in vivo). We identified that the fractions containing 10-gingerol suppressed osteoclastogenesis in RAW264.7 cells detected by tartrate-resistant acid phosphatase (TRAP) staining. In zebrafish, GHE and 10-gingerol suppressed osteoclastogenesis in prednisolone-induced osteoporosis regenerated scales to promote normal regeneration. Gene expression analysis revealed that 10-gingerol suppressed osteoclast markers in RAW264.7 cells [osteoclast-associated immunoglobulin-like receptor, dendrocyte-expressed seven transmembrane protein, and matrix metallopeptidase-9 (Mmp9)] and zebrafish scales [osteoclast-specific cathepsin K (CTSK), mmp2, and mmp9]. Interestingly, nuclear factor of activated T-cells cytoplasmic 1, a master transcription regulator of osteoclast differentiation upstream of the osteoclastic activators, was downregulated in zebrafish scales but showed no alteration in RAW264.7 cells. In addition, 10-gingerol inhibited CTSK activity under cell-free conditions. This is the first study, to our knowledge, that has found that 10-gingerol in GHE could suppress osteoclastic activity in both in vitro and in vivo conditions.

Liqing Zang, Kazuhiro Kagotani, Hiroko Nakayama, Jacky Bhagat, Yuki Fujimoto, Akihito Hayashi, Ryoji Sono, Hirotaka Katsuzaki, Norihiro Nishimura, Yasuhito Shimada. Frontiers in Cell and Developmental Biology, 9, 2021.