Transcriptome and metabolite analyses in Azadirachta indica: identification of genes involved in biosynthesis of bioactive triterpenoids

Bhambhani S, Lakhwani D, Gupta P, et al. Transcriptome and metabolite analyses in Azadirachta indica: identification of genes involved in biosynthesis of bioactive triterpenoids. SCIENTIFIC REPORTS. 2017;7(1): 5043.Azadirachta indica A. Juss, commonly known as Neem, is the reservoir of triterpenoids of economic importance. Metabolite analysis of different developmental stages of leaf and fruit suggests tissue-specific accumulation of the major triterpenoids in this important tree. Though biosynthesis of these complex molecules requires substrate flux from the isoprenoid pathway, enzymes involved in late biosynthetic steps remain uncharacterized. We established and analyzed transcriptome datasets from leaf and fruit and identified members of gene families involved in intermediate steps of terpenoid backbone biosynthesis and those related to secondary transformation leading to the tissue-specific triterpenoid biosynthesis. Expression analysis suggests differential expression of number of genes between leaf and fruit and probable participation in the biosynthesis of fruit-specific triterpenoids. Genome-wide analysis also identified members of gene families putatively involved in secondary modifications in late biosynthetic steps leading to the synthesis of highly oxygenated triterpenoids. Expression and molecular docking analyses suggest involvement of specific members of CYP450 family in secondary modifications for the biosynthesis of bioactive triterpenoids. This study generated rich genomic resource and identified genes involved in biosynthesis of important molecules, which will aid in the advancement of tools for functional genomics and elucidation of the biosynthesis of triterpenoid from this important tree

Derangement of liver function tests in dengue patients

Background: Changes in the liver function test may serve as an early marker for timely diagnosis and identification of patients who may develop severe dengue. The purpose of this study was to examine the link between dengue fever severity and liver function test. Methods: This prospective observational study was conducted in the Department of General Medicine, Madhesh Institute of health sciences, provincial hospital, Janakpurdham in which we included dengue positive patients (aged 18 years or more) based on NS1 antigen or high titer on IgM/IgG testing from July 2023 till August 2023. We excluded patients with diseases like malaria, cirrhosis of liver, enteric fever, viral hepatitis or any other disease or taking any medication which can derange LFT. Results: We included 96 patients fulfilling the study criteria. Of these, 71% had DF, 22% had DHF and 7% had DSS. Among liver enzymes, mean AST of the patients was significantly higher in DSS group of patients (775.19±39.65 U/l), as compared to those in the DF and DHF group of patients, p value <0.01. Similarly, mean ALT of the patients was significantly higher in DSS group of patients (387.8±18.6 U/l), as compared to those in the DF and DHF group of patients, p value<0.01. On the contrary, mean alkaline phosphatase levels were similar between the three patient groups. Conclusions: Based on the results our study, we conclude that raised AST and ALT levels were significantly associated with severity of DSS and DHF. Patients with dengue infection should have a baseline liver function test and subsequent LFT monitoring to detect early hepatic impairment

A protective role for nitric oxide and salicylic acid for arsenite phytotoxicity in rice (Oryza sativa L.)

The authors are thankful to Director, CSIR-National Botanical Research Institute (CSIR-NBRI), Lucknow for the facilities and for the financial support from the network projects (CSIR-INDEPTH), New Delhi, India. APS is thankful to CSIR New Delhi, India respectively, for the award of Research Associateship. RDT is gratefully thankful to Award of Emeritus Scientist (CSIR). GD is thankful to SERB-DST, New Delhi for award of NPDF. AK is thankful to UGC for award of DSKPDF. Award of Fast Track Scientist to SM from DST is gratefully acknowledged. We are also thankful to Mr. Dilip Chakraborty for technical assistance.Peer reviewedPostprin

X-ray-induced acoustic computed tomography (XACT) imaging with single-shot nanosecond x-ray

X-ray-induced acoustic computed tomography (XACT) has emerged as a promising imaging modality with broad applications in both biomedicine and nondestructive testing. The previous XACT imaging systems require thousands of averages to achieve reasonable images. Here, we report the experimental demonstration of single-shot XACT imaging of a metal object using a single-shot 50 ns x-ray pulse. A two-stage dedicated amplification and a 128-channel parallel data acquisition configuration were introduced for XACT imaging to enable sufficient acoustic signal amplification and maintain an overall low noise level for single-shot XACT imaging. Details of the system design are presented; the improved signal-to-noise ratio (>23 dB) and image reconstruction have been demonstrated with a ring ultrasound transducer array imaging system. The study paves the way for realizing real-time XACT imaging and its potential applications in image-guided intervention

Simulation study of protoacoustics as a real‐time in‐line dosimetry tool for FLASH proton therapy

BackgroundApplying ultra-high dose rates to radiation therapy, otherwise known as FLASH, has been shown to be just as effective while sparing more normal tissue compared to conventional radiation therapy. However, there is a need for a dosimeter that is able to detect such high instantaneous dose, particularly in vivo. To fulfill this need, protoacoustics is introduced, which is an in vivo range verification method with submillimeter accuracy.PurposeThe purpose of this work is to demonstrate the feasibility of using protoacoustics as a method of in vivo real-time monitoring during FLASH proton therapy and investigating the resulting protoacoustic signal when dose per pulse and pulsewidth are varied through multiple simulation studies.MethodsThe dose distribution of a proton pencil beam was calculated through a Monte Carlo toolbox, TOPAS. Next, the k-Wave toolbox in MATLAB was used for performing protoacoustic simulations, where the initial proton dose deposition was inputted to model acoustic propagations, which were also used for reconstructions. Simulations involving the manipulation of the dose per pulse and pulsewidth were performed, and the temporal and spatial resolution for protoacoustic reconstructions were investigated as well. A 3D reconstruction was performed with a multiple beam spot profile to investigate the spatial resolution as well as determine the feasibility of 3D imaging with protoacoustics.ResultsOur results showed consistent linearity in the increasing dose-per-pulse, even up to rates considered for FLASH. The simulations and reconstructions were performed for a range of pulsewidths from 0.1 to 10 μs. The results show the characteristics of the proton beam after convolving the protoacoustic signal with the varying pulsewidths. 3D reconstruction was successfully performed with each beam being distinguishable using an 8 cm × 8 cm planar array. These simulation results show that measurements using protoacoustics has the potential for in vivo dosimetry in FLASH therapy during patient treatments in real time.ConclusionThrough this simulation study, the use of protoacoustics in FLASH therapy was verified and explored through observations of varying parameters, such as the dose per pulse and pulsewidth. 2D and 3D reconstructions were also completed. This study shows the significance of using protoacoustics and provides necessary information, which can further be explored in clinical settings

Model-Based 3-D X-Ray Induced Acoustic Computerized Tomography

X-ray-induced acoustic (XA) computerized tomography (XACT) is an evolving imaging technique that aims to reconstruct the X-ray energy deposition from XA measurements. Main challenges in XACT are the poor signal-to-noise ratio and limited field-of-view, which cause artifacts in the images. We demonstrate the efficacy of model-based (MB) algorithms for three-dimensional XACT and compare with the traditional algorithms. The MB algorithm is based on iterative, matrix-free approach for regularized-least-squares minimization corresponding to XACT. The matrix-free-LSQR (MF-LSQR) and the non-iterative model-backprojection (MBP) reconstructions were evaluated and compared with universal backprojection (UBP), time-reversal (TR) and fast-Fourier transform (FFT)-based reconstructions for numerical and experimental XACT datasets. The results demonstrate the capability of MF-LSQR algorithm to reduce noisy artifacts thus yielding better reconstructions. MBP and MF-LSQR algorithms perform particularly well with the experimental XACT dataset, where noise in signals significantly affects the reconstruction of the target in UBP and FFT-based reconstructions. The TR reconstruction for experimental XACT are comparable to MF-LSQR, but takes thrice as much time and filters the frequency components greater than maximum frequency supported by the grid, resulting loss of resolution. The MB algorithms are able to overcome the challenges in XACT and hence are vital for the clinical translation of XACT

Low Temperature-Enhanced Flavonol Synthesis Requires Light-Associated Regulatory Components in Arabidopsis thaliana

Plants are continuously exposed to a myriad of stresses, which lead to the formation of secondary metabolites including flavonoids. Studies suggest that low temperature exposure leads to enhanced flavonoid accumulation in Arabidopsis thaliana. In addition, flavonoid biosynthesis is regulated by light through various regulatory factors. Therefore, plants may possess the capability to integrate light and low temperature signals for survival under freezing conditions. However, the detailed molecular mechanism and the regulatory factors associated with light- and low temperature-responsive flavonoid biosynthesis remain largely unknown. Here, we report a strict requirement for light for the low temperature-enhanced flavonol biosynthesis. Low temperature-induced expression of biosynthetic genes as well as flavonol accumulation was hampered in ELONGATED HYPOCOTYL (hy5) and myb11myb111myb12 triple mutants as compared with the wild type in Arabidopsis. Overexpression of AtHY5 in the hy5 mutant restored induction of gene expression and flavonol accumulation in response to low temperature in light. Metabolite and gene expression analysis also suggests a negative role for CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1) in accumulation of flavonols in response to low temperature. Overexpression of AtMYB12 enhanced accumulation of flavonols under low temperature in a light- dependent manner. Together, our analysis suggests the requirement for HY5 and flavonol-specific MYB regulatory factors for low temperature-induced flavonol synthesis

Model-Based X-Ray-Induced Acoustic Computed Tomography.

X-ray-induced acoustic computed tomography (XACT) provides X-ray absorption-based contrast with acoustic detection. For its clinical translation, XACT imaging often has a limited field of view. This can result in image artifacts and overall loss of quantification accuracy. In this article, we aim to demonstrate model-based XACT image reconstruction to address these problems. An efficient matrix-free implementation of the regularized LSQR (MF-LSQR)-based minimization scheme and a noniterative model back-projection (MBP) scheme for computing XACT reconstructions have been demonstrated in this article. The proposed algorithms have been numerically validated and then used to perform reconstructions from experimental measurements obtained from an XACT setup. While the commonly used back-projection (BP) algorithm produces limited-view and noisy artifacts in the region of interest (ROI), model-based LSQR minimization overcomes these issues. The model-based algorithms also reduce the ring artifacts caused due to the nonuniformity response of the multichannel data acquisition. Using the model-based reconstruction algorithms, we are able to obtain reasonable XACT reconstructions for acoustic measurements of up to 120° view. Although the MBP is more efficient than the model-based LSQR algorithm, it provides only the structural information of the ROI. Overall, it has been demonstrated that the model-based image reconstruction yields better image quality for XACT than the standard BP. Moreover, the combination of model-based image reconstruction with different regularization methods can solve the limited-view problem for XACT imaging (in many realistic cases where the full-view dataset is unavailable), and hence pave the way for future clinical translation