Publications in paediatric critical care medicine: Trends from Pakistan.

Objective: To analyse the characteristics of research published from Pakistan on paediatric critical care medicine. METHODS: The exploratory study was conducted at the Aga Khan University, Karachi from July 2021 to March 2022, and comprised a comprehensive search on MedLine, Google Scholar and PakMediNet databases for literature from Pakistan pertaining to paediatric critical care medicine published between January 2010 and December 2021. The search was done using appropriate key words. Conference abstracts and papers authored by paediatric intensivists with unrelated topics were excluded. Data was extracted on a structured spreadsheet, and was subjected to bibliometric analysis. Data was analysed using SPSS 20. RESULTS: Of the 7,514 studies identified, 146(1.94%) were analysed. These were published in 51 journals with a frequency of 13.3 per year. There were 107(73.3%) original articles, 96(65.8%) were published in PubMed-indexed journals, and 35(24%) were published in locally indexed journals. Further, 100(69.4%) papers were published from 5 paediatric intensive care units in Karachi, and 81(56%) were contributed by a single private-sector hospital. The total citation count was 1072, with 2(1.4%) papers receiving >50 citations. There was a linear trend with some skewing and an annual growth rate of >15%. Conclusion: Publications from Pakistan related to paediatric critical care medicine showed positive linear growth. There was a paucity of multicentre studies, randomised controlled trials, and high-impact publications.

Cobalt-doped zinc oxide based memristors with nociceptor characteristics for bio-inspired technology.

Neuromorphic computing is a new field of information technology, which is inspired by the biomimetic properties of the memristor as an electronic synapse and neuron. If there are electronic receptors that can transmit exterior impulses to the internal nervous system, then the use of memristors can be expanded to artificial nerves. In this study, a layer type memristor is used to build an artificial nociceptor in a very feasible and straightforward manner. An artificial nociceptor is demonstrated here through the fabrication and characterization of a cobalt-doped zinc oxide (CZO)/Au based memristor. In order to increase threshold switching performance, the surface effects of the CZO layer are eliminated by adding cobalt cobalt-doped zinc oxide (CZO) layer between the P++-Si and Au electrodes. Allodynia, hyperalgesia, threshold, and relaxation are the four distinct nociceptive behaviours that the device displays based on the strength, rate of relapse, and duration of the external stimuli. The electrons that are trapped in or released from the CZO layer's traps are responsible for these nociceptive behaviours. A multipurpose nociceptor performance is produced by this type of CZO-based device, which is crucial for artificial intelligence system applications such as neural integrated devices with nanometer-sized characteristics.

Intraventricular Lavage vs External Ventricular Drainage for Intraventricular Hemorrhage: A Randomized Clinical Trial.

Importance: Intraventricular lavage has been proposed as a minimally invasive method to evacuate intraventricular hemorrhage. There is little evidence to support its use. Objective: To evaluate the safety and potential efficacy of intraventricular lavage treatment of intraventricular hemorrhage. Design, Setting, and Participants: This single-blinded, controlled, investigator-initiated 1:1 randomized clinical trial was conducted at Aarhus University Hospital and Odense University Hospital in Denmark from January 13, 2022, to November 24, 2022. Follow-up duration was 90 days. The trial was set to include 58 patients with intraventricular hemorrhage. Prespecified interim analysis was performed for the first 20 participants. Data were analyzed from February to April 2023. Interventions: Participants were randomized to receive either intraventricular lavage or standard drainage. Main Outcomes and Measures: The main outcome was risk of catheter occlusions. Additional safety outcomes were catheter-related infections and procedure time, length of stay at the intensive care unit, duration of treatment, and 30-day mortality. The main outcome of the prespecified interim analysis was risk of severe adverse events. Efficacy outcomes were hematoma clearance, functional outcome, overall survival, and shunt dependency. Results: A total of 21 participants (median [IQR] age, 67 [59-82] years; 14 [66%] male) were enrolled, with 11 participants randomized to intraventricular lavage and 10 participants randomized to standard drainage; 20 participants (95%) had secondary intraventricular hemorrhage. The median (IQR) Graeb score was 9 (5-11), and the median (IQR) Glasgow Coma Scale score was 6.5 (4-8). The study was terminated early due to a significantly increased risk of severe adverse events associated with intraventricular lavage at interim analysis (risk difference for control vs intervention, 0.43; 95% CI, 0.06-0.81; P = .04; incidence rate ratio for control vs intervention, 6.0; 95% CI, 1.38-26.1; P = .01). The rate of catheter occlusion was higher for intraventricular lavage compared with drainage (6 of 16 patients [38%] vs 2 of 13 patients [7%]; hazard ratio, 4.4 [95% CI, 0.6-31.2]; P = .14), which met the prespecified α = .20 level. Median (IQR) procedure time for catheter placement was 53.5 (33-75) minutes for intraventricular lavage vs 12 (4-20) minutes for control (P < .001). Conclusions and Relevance: This randomized clinical trial of intraventricular lavage vs standard drainage found that intraventricular lavage was encumbered with a significantly increased number of severe adverse events. Caution is recommended when using the device to ensure patient safety. Trial Registration: ClinicalTrials.gov Identifier: NCT05204849.

Nitric oxide confers cadmium tolerance in fragrant rice by modulating physio-biochemical processes, yield attributes, and grain quality traits.

Cadmium (Cd) stress causes serious disruptions in plant metabolism, physio-biochemical processes, crop yield, and quality characters. Nitric oxide (NO) improves the quality features and nutritional contents of fruit plants. However, how NO confers Cd toxicity in fragrant rice plants, is sparse. Hence, the present study investigated the effects of 50 µM NO donor sodium nitroprusside (SNP) on physio-biochemical processes, plant growth attributes, grain yield, and quality traits of fragrant rice under Cd stress (100 mg kg-1 soil). The results revealed that Cd stress diminished rice plant growth, impaired photosynthetic apparatus and antioxidant defense system, and deteriorated the grain quality traits. However, foliar application of SNP mitigated Cd stress by improving plant growth and gas exchange attributes. Higher electrolyte leakage (EL) was accompanied with elevated levels of malondialdehyde (MDA) and hydrogen peroxide (H2O2) under Cd stress; however, exogenous application of SNP reduced them. The activities and relative expression levels of enzymatic antioxidants; superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) and non-enzymatic antioxidants, glutathione (GSH) contents were reduced by Cd stress, while SNP application regulated their activity and transcript abundances. SNP application improved fragrant rice grain yield and 2-acetyl-1-pyrroline content by 57.68 % and 75.54 % respectively, which is concomitant with higher biomass accumulation, photosynthetic efficiency, photosynthetic pigment contents, and an improved antioxidant defense system. Collectively, our results concluded that SNP application regulated the fragrant rice plant physio-biochemical processes, yield traits and grain quality characters under Cd-affected soil.

Long-term exposure changes the environmentally relevant bis(2-ethylhexyl) phthalate to be a neuro-hazardous substance disrupting neural homeostasis in emotional and cognitive functions.

Bis(2-ethylhexyl) phthalate (DEHP) is the most used member of the phthalate class of compounds. Extensive use of this plasticizer allows daily exposure to humans via various routes. A positive relationship between DEHP exposure and neurobehavioral disorders is suspected. But, there are insufficient data on the harmfulness of neurobehavioral disorders caused by DEHP exposure, particularly at daily exposure levels. In this study, we assessed the consequences of daily DEHP ingestion (2 and 20 mg/kg diets) in male mice for at least 100 days and examined its effects on neuronal functions associated with neurobehavioral disorders, such as depression and cognitive decline. We found the marked depressive behaviors and reduced learning and memory function in the DEHP-ingestion groups, and that biomarkers related to chronic stress were increased in plasma and brain tissues. Long-term DEHP ingestion induced collapse of glutamate (Glu) and glutamine (Gln) homeostasis as a result of disruption of the Glu-Gln cycle in the medial prefrontal cortex and hippocampus. The reduced glutamatergic neurotransmission activity caused by DEHP ingestion was demonstrated using an electrophysiological method. This study revealed that long-term exposure to DEHP is hazardous and can cause neurobehavioral disorders, even at daily exposure levels.

Bioengineering Techniques to Improve Nitrogen Transformation and Utilization: Implications for Nitrogen Use Efficiency and Future Sustainable Crop Production.

Nitrogen (N) is crucial for plant growth and development, especially in physiological and biochemical processes such as component of different proteins, enzymes, nucleic acids, and plant growth regulators. Six categories, such as transporters, nitrate absorption, signal molecules, amino acid biosynthesis, transcription factors, and miscellaneous genes, broadly encompass the genes regulating NUE in various cereal crops. Herein, we outline detailed research on bioengineering modifications of N metabolism to improve the different crop yields and biomass. We emphasize effective and precise molecular approaches and technologies, including N transporters, transgenics, omics, etc., which are opening up fascinating opportunities for a complete analysis of the molecular elements that contribute to NUE. Moreover, the detection of various types of N compounds and associated signaling pathways within plant organs have been discussed. Finally, we highlight the broader impacts of increasing NUE in crops, crucial for better agricultural yield and in the greater context of global climate change.

Threshold switching in nickel-doped zinc oxide based memristor for artificial sensory applications.

Electronic devices featuring biomimetic behaviour as electronic synapses and neurons have motivated the emergence of a new era in information and humanoid robotics technologies. In the human body, a nociceptor is a unique sensory neuron receptor that is capable of detecting harmful signals, leading to the central nervous system initiating a motor response. Herein, a nickel-doped zinc oxide (NZO)/Au based memristor is fabricated for the first time and characterized for artificial nociceptor application. For this, the introduction of a nickel-doped zinc oxide (NZO) layer between P++-Si and Au electrodes is used to eliminate the surface effects of the NZO layer, resulting in improved volatile threshold switching performance. Depending on the intensity, duration, and repetition rate of the external stimuli, this newly created memristor exhibits various critical nociceptive functions, including threshold, relaxation, allodynia, and hyperalgesia. The electron trapping/detrapping to/from the traps in the NZO layer is responsible for these nociceptive properties. This kind of NZO-based device produces a multifunctional nociceptor performance that is essential for applications in artificial intelligence systems, such as neural integrated devices with nanometer-sized features.

Data-driven nonstationary signal decomposition approaches: a comparative analysis.

Signal decomposition (SD) approaches aim to decompose non-stationary signals into their constituent amplitude- and frequency-modulated components. This represents an important preprocessing step in many practical signal processing pipelines, providing useful knowledge and insight into the data and relevant underlying system(s) while also facilitating tasks such as noise or artefact removal and feature extraction. The popular SD methods are mostly data-driven, striving to obtain inherent well-behaved signal components without making many prior assumptions on input data. Among those methods include empirical mode decomposition and variants, variational mode decomposition and variants, synchrosqueezed transform and variants and sliding singular spectrum analysis. With the increasing popularity and utility of these methods in wide-ranging applications, it is imperative to gain a better understanding and insight into the operation of these algorithms, evaluate their accuracy with and without noise in input data and gauge their sensitivity against algorithmic parameter changes. In this work, we achieve those tasks through extensive experiments involving carefully designed synthetic and real-life signals. Based on our experimental observations, we comment on the pros and cons of the considered SD algorithms as well as highlighting the best practices, in terms of parameter selection, for the their successful operation. The SD algorithms for both single- and multi-channel (multivariate) data fall within the scope of our work. For multivariate signals, we evaluate the performance of the popular algorithms in terms of fulfilling the mode-alignment property, especially in the presence of noise.

An Overview on Rumex dentatus L.: Its Functions as a Source of Nutrient and Health-Promoting Plant.

Rumex dentatus L. (Polygonaceae), also known as toothed dock or Aegean dock, is a medicinal plant with a high culinary value in addition to being used as an ethnomedicinal plant. This review focuses on the botanical, nutritional, phytochemical, and pharmacological activities of R. dentatus, as well as the future prospects for systematic investigations into these areas. R. dentatus has been subjected to scientific evaluation, which has confirmed its traditional uses and demonstrated a wide range of biological and pharmacological potentials, including antioxidant, anticancer, antifungal, antibacterial, anti-inflammatory, and other biological properties. Phytochemical analyses showed the presence of anthraquinones, chromones, flavonoids, and essential oils. As a result of this current review, the medicinal significance of R. dentatus has been confirmed, and future research on its unexplored aspects, such as the identification of pharmacologically active chemical constituents and related mechanisms and safety, may be stimulated, with the goal of developing it into a drug.

Molecular cloning, characterization and expression analysis of two 12-oxophytodienoate reductases (NtOPR1 and NtOPR2) from Nicotiana tabacum.

BACKGROUND: 12-oxophytodienoic acid (OPDA) is a signaling molecule involved in defense and stress responses in plants. 12-oxophytodienoate reductase (OPR) is involved in the biosynthesis of jasmonic acid and trigger the conversion of OPDA into 3-oxo-2(2'[Z]-pentenyl)-cyclopentane-1-octanoic acid (OPC-8:0). METHODS AND RESULTS: Sequence analysis revealed that Nicotiana tabacum 12-oxophytodienoate reductase 1 (OPR1) and OPR2 encoded polypeptides of 375 and 349 amino acids with molecular masses of 41.67 and 39.04 kilodaltons (kDa), respectively, while the deduced protein sequences of NtOPR1 and NtOPR2 showed high homology with other 12-oxophytodienoate reductases. BLAST (Basic local alignment search tool) analysis revealed that both NtOPRs belong to the family of Old Yellow Enzymes (OYE), and analysis of genomic DNA structure indicated that both genes include 5 exons and 4 introns. Phylogenetic analysis using MEGA X showed that NtOPR1 and NtOPR2 shared a close evolutionary relationship with Nicotiana attenuata 12-oxophytodienoate reductases. In silico analysis of subcellular localization indicated the probable locations of NtOPR1 and NtOPR2 to be the cytoplasm and the peroxisome, respectively. Tissue-specific expression assays via qRT-PCR revealed that NtOPR1 and NtOPR2 genes were highly expressed in Nicotiana tabacum roots, temperately expressed in leaves and flowers, while low expression was observed in stem tissue. CONCLUSIONS: Presently, two 12-oxophytodienoate reductase genes (NtOPR1 and NtOPR2) were cloned and comprehensively characterized. Our findings provide comprehensive analyses that may guide future deep molecular studies of 12-oxophytodienoate reductases in Nicotiana tabacum.

Comprehensive Detoxification Mechanism Assessment of Red Imported Fire Ant (Solenopsis invicta) against Indoxacarb.

The red imported fire ant (Solenopsis invicta) is one of the deadliest invasive ant species that threatens the world by disrupting biodiversity, important functions within a natural ecosystem, and community structure. They are responsible for huge economic losses in the infested countries every year. Synthetic insecticides, especially indoxacarb, have been broadly used to control S. invicta for many years. However, the biochemical response of S. invicta to indoxacarb remains largely undiscovered. Here, we used the sublethal doses of indoxacarb on the S. invicta collected from the eight different cities of Southern China. The alteration in the transcriptome profile of S. invicta following sublethal dosages of indoxacarb was characterized using high-throughput RNA-seq technology. We created 2 libraries, with 50.93 million and 47.44 million clean reads for indoxacarb treatment and control, respectively. A total of 2018 unigenes were regulated after insecticide treatment. Results indicated that a total of 158 differentially expressed genes (DEGs) were identified in the indoxacarb-treated group, of which 100 were significantly upregulated and 58 were downregulated, mostly belonging to the detoxification enzymes, such as AChE, CarE, and GSTs. Furthermore, results showed that most of these DEGs were found in several KEGG pathways, including steroid biosynthesis, other drug metabolizing enzymes, glycerolipid metabolism, chemical carcinogenesis, drug-metabolizing cytochrome P450, glutathione metabolism, glycerophospholipid metabolism, glycolysis/gluconeogenesis, and metabolism of xenobiotics. Together, these findings indicated that indoxacarb causes significant alteration in the transcriptome profile and signaling pathways of S. invicta, providing a foundation for further molecular inquiry.

Genome wide analysis of DWARF27 genes in soybean and functional characterization of GmD27c reveals eminent role of strigolactones in rhizobia interaction and nodulation in Glycine max.

BACKGROUND: Strigolactones (SLs) are newly identified hormones and their biosynthesis is stimulated under phosphate deprivation and accomplished by the action of several enzymes, including the beta-carotene isomerase DWARF27 (D27). Expression of D27 is well renowned to respond to phosphate insufficiency. However, the identification and functional analysis of the carotenoid isomerase D27 genes are not elucidated in soybean. METHODS AND RESULTS: A total of six D27 genes were identified in the soybean genome and designated on the basis of chromosomal localization. According to the findings, these genes were irregularly distributed on chromosomes, and segmental repetition led to the expansion of the soybean GmD27 gene family. Based on a neighbor-joining phylogenetic tree, the predicted D27 proteins of soybean were divided into three clades. Based on RNA seq data analysis, GmD27 genes were differently expressed in various tissues but GmD27c was the highest. Therefore, GmD27c was chosen for the additional functional study due to its rather obvious transcription in nodulation and roots. RT-qPCR results showed that GmD27c was highly expressed in different nodule stages and in response to rhizobia infection. Functional characterization of GmD27c revealed that overexpression of GmD27c led to higher nodule number, while GmD27c knockdown caused fewer nodules compared to GUS control. Furthermore, GmD27c overexpressed and knockdown lines oppositely regulated the expression of numerous nodulation genes, which are vital for the development of nodules. CONCLUSION: This study not only discovered that SL biosynthesis and signaling pathway genes are conserved, but it also revealed that SL biosynthesis gene GmD27c and legume rhizobia have close interactions in controlling plant nodule number.

Harmony but Not Uniformity: Role of Strigolactone in Plants.

Strigolactones (SLs) represent an important new plant hormone class marked by their multifunctional roles in plants and rhizosphere interactions, which stimulate hyphal branching in arbuscular mycorrhizal fungi (AMF) and seed germination of root parasitic plants. SLs have been broadly implicated in regulating root growth, shoot architecture, leaf senescence, nodulation, and legume-symbionts interaction, as well as a response to various external stimuli, such as abiotic and biotic stresses. These functional properties of SLs enable the genetic engineering of crop plants to improve crop yield and productivity. In this review, the conservation and divergence of SL pathways and its biological processes in multiple plant species have been extensively discussed with a particular emphasis on its interactions with other different phytohormones. These interactions may shed further light on the regulatory networks underlying plant growth, development, and stress responses, ultimately providing certain strategies for promoting crop yield and productivity with the challenges of global climate and environmental changes.

Conserved and Diversified Mechanism of Autophagy between Plants and Animals upon Various Stresses.

Autophagy is a highly conserved degradation mechanism in eukaryotes, executing the breakdown of unwanted cell components and subsequent recycling of cellular material for stress relief through vacuole-dependence in plants and yeast while it is lysosome-dependent in animal manner. Upon stress, different types of autophagy are stimulated to operate certain biological processes by employing specific selective autophagy receptors (SARs), which hijack the cargo proteins or organelles to the autophagy machinery for subsequent destruction in the vacuole/lysosome. Despite recent advances in autophagy, the conserved and diversified mechanism of autophagy in response to various stresses between plants and animals still remain a mystery. In this review, we intend to summarize and discuss the characterization of the SARs and their corresponding processes, expectantly advancing the scope and perspective of the evolutionary fate of autophagy between plants and animals.

Pharmacokinetic Evaluation of a Novel Donepezil-Loaded Dissolving Microneedle Patch in Rats.

Research on the development of dissolving microneedles (DMNs) has focused on bolus drug delivery, with little attention on sustained release. Here, we evaluated the sustained release, absorption pattern, and effective drug permeation of a novel donepezil-loaded DMN patch through an in vivo investigation on rats. The applications of DMN patches to the shaved skin of rats for 1 week and 1 h were compared with oral donepezil administration to assess their sustained release capabilities. We used a validated liquid chromatography-tandem mass spectrometry method to quantify donepezil in the plasma. We found that the microneedle arrays effectively delivered donepezil across the skin, with dissolution observed within 1 h of application. Furthermore, skin irritation test showed that the patches produced no irritation response. The DMN arrays also effectively increased drug permeation and demonstrated sustained release and absorption of donepezil from DMN patches. These patches allow extended dosing intervals, reduced gastrointestinal adverse effects, and convenient self-administration to mitigate poor drug compliance, making them beneficial for the treatment of elderly patients with Alzheimer's disease.

Partitioned Stator Switched Flux Machine: Novel Winding Configurations.

Torque density is one of the major limiting factors in machine design. In this paper, we propose the hybrid excited partitioned stator switched flux machine3 (HE-PSSFM3). In HE-PSSFM3, armature winding is positioned on the outer stator whereas the permanent magnet (PM) and field winding are placed at the inner stator, while the rotor is free from excitation sources and armature winding. Moreover, concentrated field winding is replaced by toroidal winding. The power splitting ratio between two stators/rotor pole combinations is analytically optimized and are validated through genetic algorithm (GA) in order to enhance average torque and flux regulation capability. The electromagnetic characteristics of the improved and initial design are evaluated and compared with existing designs, i.e., HE-PSSFM1 and HE-PSSFM2. The proposed HE-PSSFM3 has achieved high average torque, i.e., 2.0015 Nm, at same armature and field current densities of 5 A/mm2. The results show that the average torques of the proposed design are 35% and 15% greater than HE-PSSFM1 and HE-PSSFM2, respectively. Furthermore, the analysis of various parameters such as flux linkage, flux regulation, electromagnetic performances, cogging torque, back EMF, electromagnetic torque, and torque ripples are investigated using two dimensional (2D) finite element analysis (FEA). Moreover, the simulation results of the proposed design are validated through GA and analytical modeling.

Effect of Rumex Acetosa Extract, a Herbal Drug, on the Absorption of Fexofenadine.

Herbal drugs are widely used for the auxiliary treatment of diseases. The pharmacokinetics of a drug may be altered when it is coadministered with herbal drugs that can affect drug absorption. The effects of herbal drugs on absorption must be evaluated. In this study, we investigated the effects of Rumex acetosa (R. acetosa) extract on fexofenadine absorption. Fexofenadine was selected as a model drug that is a substrate of P-glycoprotein (P-gp) and organic anion transporting polypeptide 1A2 (OATP1A2). Emodine-the major component of R. acetosa extract-showed P-gp inhibition in vitro and in vivo. Uptake of fexofenadine via OATP1A2 was inhibited by R. acetosa extract in OATP1A2 transfected cells. A pharmacokinetic study showed that the area under the plasma concentration-time curve (AUC) of fexofenadine was smaller in the R. acetosa extract coadministered group than in the control group. R. acetosa extract also decreased aqueous solubility of fexofenadine HCl. The results of this study suggest that R. acetosa extract could inhibit the absorption of certain drugs via intervention in the aqueous solubility and the drug transporters. Therefore, R. acetosa extract may cause drug interactions when coadministered with substrates of drug transporters and poorly water-soluble drugs, although further clinical studies are needed.

GmMAX2-D14 and -KAI interaction-mediated SL and KAR signaling play essential roles in soybean root nodulation.

Despite of important functions of strigolactones (SLs) and karrikins (KARs) in plant development, plant-parasite and plant-fungi interactions, their roles in soybean-rhizobia interaction remain elusive. SL/KAR signaling genes GmMAX2a, GmD14s, and GmKAIs are activated by rhizobia infection. GmMAX2a restored atmax2 root hair defects and soybean root hairs were changed in GmMAX2a overexpression (GmMAX2a-OE) or knockdown (GmMAX2a-KD) mutants. GmMAX2a-KD gave fewer, whereas GmMAX2a-OE produced more nodules than GUS hairy roots. Mutation of GmMAX2a in its KD or OE transgenic hairy roots affected the rhizobia infection-induced increases in early nodulation gene expression. Both mutant hairy roots also displayed the altered auxin, jasmonate and abscisic acid levels, as further verified by transcriptomic analyses of their synthetic genes. Overexpression of an auxin synthetic gene GmYUC2a also affected SL and KAR signaling genes. GmMAX2a physically interacted with SL/KAR receptors GmD14s, GmKAIs, and GmD14Ls with different binding affinities, depending on variations in the critical amino acids, forming active D14/KAI-SCFMAX2 complexes. The knockdown mutant roots of the nodule-specifically expressing GmKAIs and GmD14Ls gave fewer nodules, with altered expression of several early nodulation genes. The expression levels of GmKAIs, and GmD14Ls were markedly changed in GmMAX2a mutant roots, so did their target repressor genes GmD53s and GmSMAX1s. Thus, SL and KAR signaling were involved in soybean-rhizobia interaction and nodulation partly through interactions with hormones, and this may explain the different effects of MXA2 orthologs on legume determinate and indeterminate nodulation. The study provides fresh insights into the roles of GmMAX2-mediated SL/KAR signaling in soybean root hair and nodule formation.

De novo transcriptome sequencing and metabolite profiling analyses reveal the complex metabolic genes involved in the terpenoid biosynthesis in Blue Anise Sage (Salvia guaranitica L.).

Many terpenoid compounds have been extracted from different tissues of Salvia guaranitica. However, the molecular genetic basis of terpene biosynthesis pathways is virtually unknown. In this study, approximately 4 Gb of raw data were generated from the transcriptome of S. guaranitica leaves using Illumina HiSeq 2000 sequencing. After filtering and removing the adapter sequences from the raw data, the number of reads reached 32 million, comprising 186 million of high-quality nucleotide bases. A total of 61,400 unigenes were assembled de novo and annotated for establishing a valid database for studying terpenoid biosynthesis. We identified 267 unigenes that are putatively involved in terpenoid metabolism (including, 198 mevalonate and methyl-erythritol phosphate (MEP) pathways, terpenoid backbone biosynthesis genes and 69 terpene synthases genes). Moreover, three terpene synthase genes were studied for their functions in terpenoid biosynthesis by using transgenic Arabidopsis; most transgenic Arabidopsis plants expressing these terpene synthetic genes produced increased amounts of terpenoids compared with wild-type control. The combined data analyses from the transcriptome and metabolome provide new insights into our understanding of the complex metabolic genes in terpenoid-rich blue anise sage, and our study paves the way for the future metabolic engineering of the biosynthesis of useful terpene compounds in S. guaranitica.

A Simple and Sensitive Liquid Chromatography with Tandem Mass Spectrometric Method for the Simultaneous Determination of Anthraquinone Glycosides and Their Aglycones in Rat Plasma: Application to a Pharmacokinetic Study of Rumex acetosa Extract.

Rumex acetosa (R. acetosa) has been used in folk remedies for gastrointestinal disorders and cutaneous diseases. Rumex species, in particular, contain abundant anthraquinones. Anthraquinone glycosides and aglycones show different bioactive effects. However, information on the pharmacokinetics of anthraquinone glycosides is limited, and methods to quantify anthraquinone glycosides in plasma are rarely available. A simple and sensitive liquid chromatography-tandem mass spectrometric bioanalytical method for the simultaneous determination of both anthraquinone glycosides and their aglycones, including emodin, emodin-8-O-ß-d-glucoside, chrysophanol, chrysophanol-8-O-ß-d-glucoside, physcion, and physcion-8-O-ß-d-glucoside , in a low volume of rat plasma (20 µL) was established. A simple and rapid sample preparation was employed using methanol as a precipitating agent with appropriate sensitivity. Chromatographic separation was performed on HPLC by using a biphenyl column with a gradient elution using 2 mM ammonium formate (pH 6) in water and 2 mM ammonium formate (pH 6) in methanol within a run time of 13 min. The anthraquinones were detected on triple-quadrupole mass spectrometer in negative ionization mode using multiple-reaction monitoring. The method was validated in terms of selectivity, linearity, accuracy, precision, recovery, and stability. The values of the lower limit of quantitation of anthraquinones were 1⁻20 ng/mL. The intra-batch and inter-batch accuracies were 96.7⁻111.9% and the precision was within the acceptable limits. The method was applied to a pharmacokinetic study after oral administration of R. acetosa 70% ethanol extract to rats at a dose of 2 g/kg.