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CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated protein) system has emerged as a popular toolkit to edit specific regions of the genome in several taxa. A 20 bp single guide RNA (sgRNA) directs the Cas 9 endonuclease enzyme to a precise location in the genome for double stranded DNA cleavage and further repair, predominantly through Non-homologous End Joining (NHEJ), resulting in addition or deletions. In this regard, the dsDNA cleavage by Cas9 depends mainly on various factors in sgRNA design. Current web-based tools for sgRNA design suggest a wide variety of factors to choose a suitable sgRNA. Therefore, it is important to check the cleaving efficiency of the Cas9 + sgRNA complex called Ribonucleoprotein complex through in vitro restriction assay before proceeding with microinjection to achieve in vivo restriction of the target gene in an organism. In the present study we have demonstrated the cleaving efficiency of different sgRNAs in restricting the white gene of the Fall armyworm, Spodoptera frugiperda. [ABSTRACT FROM AUTHOR]

The CRISPR/Cas9 technology has opened up newer avenues in insect pest management like precision guided sterile insect technique (pgSIT) which achieves a highly specific mutation in the target genes such as spermatogenesis, sex determination related genes etc. In this regard, validating the loss-of-function of the target gene/s is a prerequisite before the final application. This is easily achieved through DNA-free editing by embryonic delivery of the cognate ribo nucleo protein complex (RNP) into G0 stage eggs. Obtaining uniform G0 stage eggs is necessary to offset the microinjection injury and have high heritability of the genomic edits. We to optimized a method to obtain intact eggs of Oriental fruit fly, Bactrocera dorsalis (Hendel) without injury for microinjection by modifying the oviposition behaviour of the gravid female of B. dorsalis to retrieve intact eggs and to obtain large number of G0 eggs for genome editing. This paper describes a method to obtain required number of eggs for such studies. Out of two egg laying methods, the one with a small container with water covered with parafilm and topped with a thin banana pulp slice provided intact eggs. Maximum oviposition was observed between 20-60 days after eclosion. By the present finding we can obtain sufficient eggs for microinjection at 15-minute interval. [ABSTRACT FROM AUTHOR]

The objective of this study was to develop and employ a numerical simulation strategy for predicting the liquid water content (LWC) at the test-section plane of the NASA Glean king Research Tunnel (IRT) as well as to characterize the icing cloud uniformity as a function of tunnel speed, droplet size, etc. The droplets were injected with a polydisperse distribution and based on previous computational airflow results, which included the spray-bar wakes, the air jets, and the heat-exchanger flow. To simulate the effects of turbulent diffusion behavior of water droplets in the IRT, a continuous-random-walk methodology was employed, which corrects for nonhomogeneous turbulence. To first understand the cloud dynamics emanating from isolated nozzles, experiments and simulations were conducted with only four injectors spraying in the IRT. The simulations showed good representation of the width and position (though not necessarily the shape) of the individual spray clouds at the test-section plane. Next, droplets issued by the baseline group of spray nozzles were simulated in order to determine the test-section LWC distributions which in turn were used to compare with experiments. The simulations indicated the importance of including turbulent diffusion, and that increased tunnel speed and droplet size tended to reduce overall uniformity, which was generally consistent with experimental results.

Purpose: Disparities have been reported in women treated for breast cancer (BrCa). This study examines potential disparities in BrCa treatment offered based on race and age from a multicenter radiation department. Methods and Materials: We identified 901 patients with early stage BrCa who received curative intent radiation therapy (RT) between 2004 and 2018. Data extracted included age, race, disease stage, treatment technique, treatment dates, and fractionation. Patient race was recorded as Asian, Black, Hispanic, and White. RT technique delivered was classified as a type of external beam radiation therapy or brachytherapy/intraoperative radiation therapy. Fractionation schema were defined as 1) standard fractionation, 1.8-2 Gy; 2) hypofractionation, 2.5-2.67 Gy; 3) accelerated partial breast irradiation (APBI), 3.4 Gy – 4.25 Gy, and 4) intraoperative radiation therapy, single dose of 20 Gy. Stage was recorded using TNM staging. The χ2 test and a multivariable multinomial logistic regression model were used to assess whether patient characteristics, such as age, race, or stage influenced fractionation schemes. Results with 2-sided P values < .05 were considered statistically significant. Results: Racial composition of the study was 13.8% Asian, 22% Black, 29%, White, and 35.1% Hispanic. Mean age was 61 and was divided into 4 age range groups: 30 to 49 (n = 160), 50 to 59 (n = 231), 60 to 69 (n = 294), and ≥70 years (n = 216). In addition, 501 patients (56%) received hypofractionation, 342 (38.8%) received standard fractionation, and 58 (7.1%) received APBI, respectively. For all groups, hypofractionation became more common over time. Age ≥70 years was associated with 9 times higher odds of APBI and 14 times higher odds of hypofractionation, compared with age 30 to 49 years. After adjusting for the other predictors in a multivariable multinomial logistic regression model, the race distribution differed among the 3 groups (P = .03), with a smaller percentage of Hispanics and higher percentage of blacks in the standard group. Conclusions: This study of a diverse cohort of patients with breast cancer failed to identify treatment differences associated by race. The study found an association between age and hypofractionation.

Reliability evaluation of distribution systems plays a crucial role in ensuring the uninterrupted supply of electricity to consumers. This paper presents a comprehensive analysis of distribution system reliability using the Failure Mode and Effects Analysis (FMEA) technique. The study focuses on evaluating the reliability of the distribution system considering various failure modes, including busbar failure, section failure, distributor failure, transformer failure, maintenance-related issues, transient failures, and the impact of weather conditions. The findings of this research can assist utilities and system operators in identifying critical failure modes and implementing appropriate measures to enhance the reliability of distribution systems, ensuring continuous and reliable electricity supply to consumers.

Sustainable development is the prime criterion for the conservation of natural resources and to keep an environmental balance between conserving natural resources such as coal, gas and oil, which are non-renewable sources of energy, it is necessary to introduce renewable sources of energy such as solar, tidal, wind and geothermal. Solar energy has shown great potential in the last two decades due to Sunlight readily available source. Different photovoltaics like organic and inorganic are developed in the previous several years for power generation in various applications. At present silicon-based solar cells are commercialized as they have shown good conversion efficiency compared to other solar cells and good stability in terms of life. However, the high price and complex fabrication process limit its application. For these reasons, researchers are looking forward to organic photovoltaic cells out of which dye-sensitized solar cell shows good efficiency, low cost of fabrication, and easy process of construction. In this paper, the cost-effective and easy fabrication process is used to fabricate four types of Dye-sensitized solar cell Dye-sensitized solar cell (DSSC) using onion peel dye sensitizer, polymethylmethacrylate (PMMA), and triiodide-based gel polymer electrolyte (GPE). All four Dye-sensitized solar cell (DSSC) have a different ratio of PMMA gel and Triiodide as liquid Electrolyte (LE). Out of these four fabricated DSSC's, the DSSC which has 80:20 liquid and gel solution provided the highest efficiency of range 11.32%. All the dye-sensitized solar cells are tested under the solar luminance of 37.288 mW/cm2. A maximum open-circuit photovoltage of 0.386 V with a fill factor of 0.439 made it the best proto-model to be used for practical applic [ABSTRACT FROM AUTHOR]

Abstract Background Barth syndrome (BTHS) is a rare genetic disease that is characterized by cardiomyopathy, skeletal myopathy, neutropenia, and growth abnormalities and often leads to death in childhood. Recently, elamipretide has been tested as a potential first disease-modifying drug. This study aimed to identify patients with BTHS who may respond to elamipretide, based on continuous physiological measurements acquired through wearable devices. Results Data from a randomized, double-blind, placebo-controlled crossover trial of 12 patients with BTHS were used, including physiological time series data measured using a wearable device (heart rate, respiratory rate, activity, and posture) and functional scores. The latter included the 6-minute walk test (6MWT), Patient-Reported Outcomes Measurement Information System (PROMIS) fatigue score, SWAY Balance Mobile Application score (SWAY balance score), BTHS Symptom Assessment (BTHS-SA) Total Fatigue score, muscle strength by handheld dynamometry, 5 times sit-and-stand test (5XSST), and monolysocardiolipin to cardiolipin ratio (MLCL:CL). Groups were created through median split of the functional scores into “highest score” and “lowest score”, and “best response to elamipretide” and “worst response to elamipretide”. Agglomerative hierarchical clustering (AHC) models were implemented to assess whether physiological data could classify patients according to functional status and distinguish non-responders from responders to elamipretide. AHC models clustered patients according to their functional status with accuracies of 60–93%, with the greatest accuracies for 6MWT (93%), PROMIS (87%), and SWAY balance score (80%). Another set of AHC models clustered patients with respect to their response to treatment with elamipretide with perfect accuracy (all 100%). Conclusions In this proof-of-concept study, we demonstrated that continuously acquired physiological measurements from wearable devices can be used to predict functional status and response to treatment among patients with BTHS.

Digital system algorithms such as FFT algorithms, convolution, image processing algorithm, etc. deploy Multiply and Accumulate (MAC) unit as an evaluative component. The efficiency of a MAC typically relies on the speed of operation, power dissipation, and chip area along with the complexity level of the circuit. In this research paper, a power-delay-efficient signed-floating-point MAC (SFMAC) is proposed using Universal Compressor based Multiplier (UCM). Instead of having a complex design architecture, a simple multiplexer-based circuit is used to achieve a signed-floating output. The 8£8 SFMAC can take 8-bit mantissa and 3-bit exponent and therefore, the input to the SFMAC can be in the range of - (7.96875)10 to +(7.96875)10. The design and implementation of the proposed architecture is executed on the Cadence Spectre tool in GPDK 90 nm and TSMC 130 nm CMOS, which proves as power and delay efficient. [ABSTRACT FROM AUTHOR]

BackgroundLimited data accuracy is often cited as a reason for caution in the integration of physiological data obtained from consumer-oriented wearable devices in care management pathways. The effect of decreasing accuracy on predictive models generated from these data has not been previously investigated. ObjectiveThe aim of this study is to simulate the effect of data degradation on the reliability of prediction models generated from those data and thus determine the extent to which lower device accuracy might or might not limit their use in clinical settings. MethodsUsing the Multilevel Monitoring of Activity and Sleep in Healthy People data set, which includes continuous free-living step count and heart rate data from 21 healthy volunteers, we trained a random forest model to predict cardiac competence. Model performance in 75 perturbed data sets with increasing missingness, noisiness, bias, and a combination of all 3 perturbations was compared to model performance for the unperturbed data set. ResultsThe unperturbed data set achieved a mean root mean square error (RMSE) of 0.079 (SD 0.001) in predicting cardiac competence index. For all types of perturbations, RMSE remained stable up to 20%-30% perturbation. Above this level, RMSE started increasing and reached the point at which the model was no longer predictive at 80% for noise, 50% for missingness, and 35% for the combination of all perturbations. Introducing systematic bias in the underlying data had no effect on RMSE. ConclusionsIn this proof-of-concept study, the performance of predictive models for cardiac competence generated from continuously acquired physiological data was relatively stable with declining quality of the source data. As such, lower accuracy of consumer-oriented wearable devices might not be an absolute contraindication for their use in clinical prediction models.

Background: a humidity sensor is used to sense and measure the relative humidity of air. A new composite system has been fabricated using environmental pollutants such as carbon black and low-cost zinc oxide, and it acts as a humidity sensor. Residual life of the sensor is calculated and an expert system is modelled. For properties and nature confirmation, characterization is performed, and a sensing material is fabricated. Methodology: characterization is performed on the fabricated material. Complex impedance spectroscopy (CIS), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscopy (SEM) are all used to confirm the surface roughness, its composite nature as well as the morphology of the composite. The residual lifetime of the fabricated humidity sensor is calculated by means of accelerated life testing. An intelligent model is designed using artificial intelligence techniques, including the artificial neural network (ANN), fuzzy inference system (FIS) and adaptive neuro-fuzzy inference system (ANFIS). Results: maximum conductivity obtained is 6.4£10-3 S/cm when zinc oxide is doped with 80% of carbon black. Conclusion: the solid composite obtained possesses good humidity-sensing capability in the range of 30-95%. ANFIS exhibits the maximum prediction accuracy, with an error rate of just 1.1%. [ABSTRACT FROM AUTHOR]

An electrolytic capacitor is a widely used, passive component in electronics and the manufacturing industry. But it is known for its low reliability when used in critical applications and environments. The prediction of the residual life of a capacitor is a challenging issue. It is beneficial for users as they will be ready to replace a component when it is faulty. This will save them replacement and repairing costs. This paper focusses on the mathematical modelling and behaviour analysis of electrolytic capacitors related to various electrical parameters and environmental stresses. A novel method is proposed to access the impact of humidity on the health of a capacitor. A capacitor's end of life is explored using the design of experiments approach. After calculating the lifetime of a capacitor, an expert system is modelled that can predict the sudden crash of the capacitor before it actually fails, using various statistical and analytical techniques. A comparison was done using the experimental calculated lifetime and proposed analytical method outcome. The proposed corrective model of lifetime calculation proves 92.76% accuracy. This shortfall is due to the fact that some electrical and environmental variables change at a pace that is too fast for the prediction system to respond. [ABSTRACT FROM AUTHOR]

Recently, an acoustic lens has been proposed for volumetric focusing as an alternative to conventional reconstruction algorithms in Photoacoustic (PA) Imaging. Acoustic lens can significantly reduce computational complexity and facilitate the implementation of real-time and cost-effective systems. However, due to the fixed focal length of the lens, the Point Spread Function (PSF) of the imaging system varies spatially. Furthermore, the PSF is asymmetric, with the lateral resolution being lower than the axial resolution. For many medical applications, such as in vivo thyroid, breast and small animal imaging, multiple views of the target tissue at varying angles are possible. This can be exploited to reduce the asymmetry and spatial variation of system the PSF with simple spatial compounding. In this article, we present a formulation and experimental evaluation of this technique. PSF improvement in terms of resolution and Signal to Noise Ratio (SNR) with the proposed spatial compounding is evaluated through simulation. Overall image quality improvement is demonstrated with experiments on phantom and ex vivo tissue. When multiple views are not possible, an alternative residual refocusing algorithm is proposed. The performances of these two methods, both separately and in conjunction, are compared and their practical implications are discussed. Keywords: Acoustic lens, Photoacoustic camera, Point Spread Function, Resolution, Refocusing, Spatial compounding

Using low-cost materials, carbon, and alum, a new solid composite electrolyte system was fabricated and characterized using various techniques. Complex impedance spectroscopy was used to measure its ionic conductivity. A maximum conductivity of 3.4×10−5 S/cm was obtained when alum was doped with 40% carbon. An Arrhenius behavior was reported when the temperature dependence of conductivity was analyzed. Scanning electron microscopy was used to study the surface morphology of the synthesized electrolyte. Fourier transform infrared and X-ray diffraction results confirmed the formation of composite material. The synthesized solid composite electrolyte exhibited excellent humidity sensing behavior in the relative humidity range of 15%–65%. Various humidity characteristics of the sample were measured such as hysteresis loop, recovery, and response time of the sensor. An expert system was modeled using artificial intelligence techniques and failure of the sensor was predicted with 97.2% accuracy using artificial neural networks.

Some of the challenges in translating photoacoustic (PA) imaging to clinical applications includes limited view of the target tissue, low signal to noise ratio and the high cost of developing real-time systems. Acoustic lens based PA imaging systems, also known as PA cameras are a potential alternative to conventional imaging systems in these scenarios. The 3D focusing action of lens enables real-time C-scan imaging with a 2D transducer array. In this paper, we model the underlying physics in a PA camera in the mathematical framework of an imaging system and derive a closed form expression for the point spread function (PSF). Experimental verification follows including the details on how to design and fabricate the lens inexpensively. The system PSF is evaluated over a 3D volume that can be imaged by this PA camera. Its utility is demonstrated by imaging phantom and an ex vivo human prostate tissue sample.

Vitellogenin (Vg), a yolk protein precursor, plays an important role in the oocyte development of insects and is an important target of genetic pest management. Vg is synthesized in the fat body, transported through haemolymph and accumulates in developing oocytes. In this regard, the eggplant shoot and fruit borer, Leucinodes orbonalis (Lepidoptera: Crambidae) is the major pest in South and South East Asia and a serious concern for farmers. Therefore, in the present study, we have cloned and characterized Vg from L. orbonalis (LoVg) for further applications. The cloned Vg consisted of 5,370 base pairs encoding 1,790 amino acid residues long protein. Further, sequence alignment revealed that LoVg has three conserved domains: a Vitellogenin N domain (LPD-N), a domain of unknown function protein families (DUF1943), and a von Willebrand factor type D domain (VWD). Using phylogenetic analysis, it was found that LoVg evolved alongside homologous proteins from different insects. The real-time expression levels of LoVg were significantly greater in female adults followed by the pupal stage. This suggests that Vg production and absorption in L. orbonalis occurs in the later pupal stage. Our studies showed that editing LoVg using CRISPR/Cas9 did not affect the total number of eggs laid but affected egg hatchability. These studies help us to design newer approaches in insect pest management through genetic suppression for sustainable pest management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In quantum field theory above two spacetime dimensions, one is usually only able to construct exact operator maps from UV to IR of strongly coupled renormalization group flows for the most symmetry-protected observables. Famous examples include maps of chiral rings in 4D N=2 supersymmetry. In this Letter, we construct the first nonperturbative UV-IR map for less protected operators: starting from a particularly "simple" UV strongly coupled non-Lagrangian 4D N=2 quantum field theory, we show that a universal nonchiral quarter-Bogomol'nyi-Prasad-Sommerfield ring can be mapped exactly and bijectively to the IR. In particular, strongly coupled UV dynamics governing infinitely many null states manifest in the IR via Fermi statistics of free gauginos. Using the concept of arc space, this bijection allows us to compute the exact UV Macdonald index in the IR.

In electrical discharge machining (EDM), the machined surface quality can be affected by the excessive temperature generation during the machining process. To achieve a longer life of the finished part, the machined surface quality plays a key role in maintaining its overall integrity. Surface roughness is an important quality evaluation of a material's surface that has considerable influence on mechanical performance of the material. Herein, a sintered cermet tooltip with 75% copper and 25% titanium carbide was used as tool electrode for processing H13 steel. The experiments have been performed to investigate the effects of EDM parameters on the machined surface roughness. The findings show that, as the pulse current, pulse length, and pulse interval are increased, the surface roughness tends to rise. The most significant determinant for surface roughness was found to be pulse current. A semi-empirical surface roughness model was created using the characteristics of the EDM technique. Buckingham's theorem was used to develop a semi-empirical surface roughness prediction model. The semi-empirical model's predictions were in good agreement with the experimental studies, and the built empirical model based on physical features of the cermet tooltip was tested using dimensional analysis.

Targeted cancer therapy provides the basis for the arrest of tumor growth in aggressive pancreatic carcinoma; however, a number of protein-based targeted toxins lack efficacy due to insufficient endosomal escape after being endocytosed. Therefore, we tested a fusion protein of the ribosome-inactivating protein dianthin and human epidermal growth factor in combination with a glycosylated triterpene (SO1861) that serves as an endosomal escape enhancer. In vitro investigations with the pancreatic carcinoma cell lines BxPC-3 and MIA PaCa-2 revealed no significant differences to off-target cells in the half maximal inhibitory concentration (IC 50 ) for the fusion protein. In contrast, combination with SO1861 decreased the IC 50 for BxPC-3 cells from 100 to 0.17 nm, whereas control cells remained unaffected. Monotherapy of BxPC-3 xenografts in CD-1 nude mice led to a 51.7% average reduction in tumor size (40.8 mm 3 ) when compared to placebo; however, combined treatment with SO1861 resulted in a more than 13-fold better efficacy (3.0 mm 3 average tumor size) with complete regression in 80% of cases. Immunohistochemical analyses showed that tumor cells with lower target receptor expression are, in contrast to the combination therapy, able to escape from the monotherapy, which finally results in tumor growth. At the effective concentration, we did not observe liver toxicity and saw no other side effects with the exception of a reversible skin hardening at the SO1861 injection site, alongside an increase in platelet counts, plateletcrit, and platelet distribution width. In conclusion, combining a targeted toxin with SO1861 is proven to be a very promising approach for pancreatic cancer treatment., (© 2017 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.)

Triterpenoidal saponins are synthesized in the roots of Saponaria officinalis L. The same plant is also a source for the toxin Saporin, which is a ribosome-inactivating protein. Triterpenoidal saponins are known to increase the cytotoxicity of Saporin by modulating its intracellular trafficking. Here, we investigated if the combinatorial effects elicited by purified saponins and Saporin can be applied to increase the therapeutic efficacy of the immunotoxin Saporin-Rituximab. First, saponins were purified by high-performance liquid chromatography. Thereafter, their intrinsic cytotoxicity was evaluated on Ramos cells with no observed effect up to 5 µg/mL, however, saponins increased the cytotoxicity of Saporin, while no influence was observed on its N -glycosidase activity. Saporin-Rituximab bound to CD20 in Ramos cells and, in the absence of saponins, had a GI 50 (concentration inhibiting cell growth to 50 %) of 7 nM. However, in the presence of a nontoxic concentration of saponins, the GI 50 of Saporin-Rituximab was 0.01 nM, a nearly 700-fold increase in efficacy. Moreover, two further immunotoxins, namely Saporin-anti-CD22 and Saporin-anti-CD25, were tested in combination with saponins yielding enhancement factors of 170-fold and 25-fold, respectively. All three receptors are present in Ramos cells and the differences in cytotoxicity enhancement may be explained by the differing expression levels of the cellular receptors. The application of purified saponins from S. officinalis L. is therefore a new strategy to potentially improve the cytotoxicity and therapeutic efficacy of Rituximab-immunotoxins for the treatment of B-cell lymphoma., (Georg Thieme Verlag KG Stuttgart · New York.)

The therapeutic relevance of immunotoxins is based on the conjugation of monoclonal antibodies to toxins. In cancer therapies, the conjugated antibodies not only direct the binding of immunotoxins to cancer-specific receptors and mediate the elimination of tumor cells through the innate immune system, but also increase target cytotoxicity by the intrinsic toxin activity. In the present study, the therapeutic antibodies Cetuximab (anti-EGFR, Erbitux(®)), Panitumumab (anti-EGFR, Vectibix(®)) and Trastuzumab (anti-HER2, Herceptin(®)) were chemically conjugated to the toxin dianthin. In the first instance, recombinant dianthin was characterized by mass spectrometry and its stability was analyzed by circular dichroism. Dianthin showed increased cytotoxicity on MCF-7 cells when tested in combination with a glycosylated triterpenoid (SO1861) in a real-time impedance-based cytotoxicity assay. In data obtained by live cell imaging, SO1861 specifically mediated the endo/lysosomal escape of dianthin without disrupting the plasma membrane. The purity of immunotoxins was confirmed by SDS-PAGE and Western blot. Their cytotoxicity was evaluated in the presence of SO1861 and dianthin-Cetuximab presented a GI50 (50% growth inhibition) of 5.3pM, dianthin-Panitumumab of 1.5pM, and dianthin-Trastuzumab of 23pM. Finally, the specificity of these immunotoxins was validated in a fluorescence-based real-time assay, where their binding to target cells was prevented by preincubation with an excess of label-free unconjugated antibody. Based on these data, we propose the use of dianthin and SO1861 as a new platform technology to enhance the efficacy of therapeutic antibodies., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objective: To study and compare the conventional extraction procedure with microwave assisted extraction (MAE) for some Ayurvedic Rasayana drugs and to evaluate their antioxidant potential and carry out the characterization of extracts by thin layer chromatography., Methods: Three Ayurvedic rasayana plants Allium sativum Linn., Bombax ceiba Linn. and Inula racemosa Hook. were evaluated for an improved MAE methodology by determining the effects of grinding degree, extraction solvent, effect of dielectric constant and duration of time on the extractive value. Antioxidant potential of all three drugs was evaluated with 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reducing power was determined by using Gallic acid as standard. Further thin layer chromatographic (TLC) analysis was performed on pre-activated Silica Gel G plates and Rf value were compared with those reported for the important biomarkers., Results: The total extractive value for Allium sativum Linn. was 36.95% (w/w) and 49.95% (w/w) for ethanol extraction respectively. In case of Bombax ceiba Linn. the yield of aqueous extract by MAE was 50% (w/w) compared to 42% (w/w) in ethanol (50% v/v). Percent yield of Inula racemosa Hook. in aqueous extract was found to be 27.55% (w/w) which was better than ethanol extract (50%) where the yield was 25.95% (w/w). Upon antioxidant activity evaluation. sativum extract showed an absorbance of 0.980±0.92 at concentration of 500 μg with maximum reducing capacity. This was followed by. ceiba Linn. 0.825±0.98 and. racemosa Hook. with 0.799±2.01 at a concentration of 500 μg. TLC based standardization of. sativum Linn. extract shows single spot with Rf value of 0.38, B. ceiba Linn. extract shows Rf values were 0.23, 0.58, 0.77, 0.92 and I. racemosa Hook. extract spot had a Rf value of 0.72., Conclusions: A significant improvement in extractive values was observed as a factor of time and other advantages by using MAE technology. All three drugs have high antioxidant potential and a TLC profiling similar to reported ones. The presence of fructan type polysaccharide can be further utilized for bioactivity directed fractionation and evaluation of immunomodulatory activity.

Protein-based therapeutics with cytosolic targets are capable of exhibiting their therapeutic effect once they have escaped from the endosomes or lysosomes. In this study, the reporters-horseradish peroxidase (HRP), Alexa Fluor 488 (Alexa) and ricin A-chain (RTA)-were investigated for their capacity to monitor the endo/lysosomal escape of the ribosome-inactivating protein, saporin. The conjugates-saporin-HRP, (Alexa)saporin and saporin-KQ-RTA-were constructed, and the endo/lysosomal escape of these conjugates alone (lack of endo/lysosomal release) or in combination with certain structurally-specific triterpenoidal saponins (efficient endo/lysosomal escape) was characterized. HRP failed in reporting the endo/lysosomal escape of saporin. Contrastingly, Alexa Fluor 488 successfully allowed the report of the process at a toxin concentration of 1000 nM. In addition, single endo/lysosome analysis facilitated the determination of the amount of (Alexa)saporin released from each vesicle. RTA was also successful in reporting the endo/lysosomal escape of the enzymatically inactive mutant, saporin-KQ, but in this case, the sensitivity of the method reached a toxin concentration of 10 nM. In conclusion, the simultaneous usage of Alexa Fluor 488 and RTA as reporters may provide the possibility of monitoring the endo/lysosomal escape of protein-based therapeutics in the concentration range of 10-1000 nM.