Fenfuro®-mediated arrest in the formation of protein-methyl glyoxal adducts: a new dimension in the anti-hyperglycemic potential of a novel fenugreek seed extract.

The fenugreek plant (Trigonella foenum-graecum) is traditionally known for its anti-diabetic properties owing to its high content of furostanolic saponins, which can synergistically treat many human ailments. Non-enzymatic protein glycation leading to the formation of Advanced Glycation End products (AGE) is a common pathophysiology observed in diabetic or prediabetic individuals, which can initiate the development of neurodegenerative disorders. A potent cellular source of glycation is Methyl Glyoxal, a highly reactive dicarbonyl formed as a glycolytic byproduct. We demonstrate the in vitro glycation arresting potential of Fenfuro®, a novel patented formulation of Fenugreek seed extract with clinically proven anti-diabetic properties, in Methyl-Glyoxal (MGO) adducts of three abundant amyloidogenic cellular proteins, alpha-synuclein, Serum albumin, and Lysozyme. A 0.25% w/v Fenfuro® was able to effectively arrest glycation by more than 50% in all three proteins, as evidenced by AGE fluorescence. Glycation-induced amyloid formation was also arrested by more than 36%, 14% and 15% for BSA, Alpha-synuclein and Lysozyme respectively. An increase in MW by attachment of MGO was also partially prevented by Fenfuro® as confirmed by SDS-PAGE analysis. Glycation resulted in enhanced aggregation of the three proteins as revealed by Native PAGE and Dynamic Light Scattering. However, in the presence of Fenfuro®, aggregation was arrested substantially, and the normal size distribution was restored. The results cumulatively indicated the lesser explored potential of direct inhibition of glycation by fenugreek seed in addition to its proven role in alleviating insulin resistance. Fenfuro® boosts its therapeutic potential as an effective phytotherapeutic to arrest Type 2 diabetes.

Fenfuro^® is a novel patented formulation of Fenugreek seed extract with more than 45% furostanolic saponins and anti-diabetic property free from any side effect as established through clinical study.In the present study, the role of Fenfuro® in arresting in vitro AGE formation and glycation-induced amyloid formation has been demonstrated with the help of three amyloidogenic proteins, namely Human Lysozyme, Human alpha-synuclein and Bovine Serum Albumin using Methyl Glyoxal as the glycating agent.A 0.25% (w/v) ethanolic solution of Fenfuro® resulted in more than 50% arrest in glycation with simultaneous prevention of aggregation as demonstrated by native PAGE, DLS and inhibition of development of Thio-T positive amyloid like entities.The studies collectively aim toward the development of a safe therapeutic method for arresting protein glycation through direct physical intervention.

Development and validation of a gel wax phantom to evaluate geometric accuracy and measurement of a hyperechoic target diameter in diagnostic ultrasound imaging.

Diagnostic ultrasound (US) scanners are generally evaluated using proprietary quality assurance (QA) phantoms, but their prohibitively high cost may prevent organizations to perform the necessary tests. This study aimed to develop a low-cost gel wax phantom with targets to determine the lateral and axial resolution and diameter of a hyperechoic target in an US scanner. The acoustic property (AP) of gel wax, which includes the speed of sound (cus), acoustic impedance (Z), and attenuation coefficient (µ), were determined for multiple transducers operating at 2.25, 5, 10, 15, and 30 MHz. These results were compared to the AP of soft tissue. Two polytetrafluoroethylene (PTFE) rectangular frames with holes separated by 5, 10, and 20 mm were constructed. Nylon filaments and stainless-steel disc (SS disc) (diameter = 16.8 mm) were threaded through the frames and suitably placed in gel wax to obtain orthogonal targets in the phantom. The target dimensions obtained from computerized tomography (CT) and US images of the phantom were compared for phantom validation. The average cus=1431.4 m/s, mass density ρ = 0.87 g/cm3, Z = 1.24 MRayls, and µ ranged from 0.7 to 0.98 dB/cm/MHz for gel wax at 22 °C. The US image measurement exhibited a maximum error in determining the diameter of the SS disc, resulting in a value of 18 mm instead of its actual value of 16.8 mm. The phantom volume decreased by 1.8% in 62 weeks. The present phantom is affordable, stable, customizable, and can be used to evaluate diagnostic US scanners across multiple centers.

Improvement of LED-based photoacoustic imaging using lag-coherence factor (LCF) beamforming.

BACKGROUND: Owing to its portability, affordability, and energy-efficiency, LED-based photoacoustic (PA) imaging is increasingly becoming popular when compared to its laser-based alternative, mainly for superficial vascular imaging applications. However, this technique suffers from low SNR and thereby limited imaging depth. As a result, visual image quality of LED-based PA imaging is not optimal, especially in sub-surface vascular imaging applications. PURPOSE: Combination of linear ultrasound (US) probes and LED arrays are the most common implementation in LED-based PA imaging, which is currently being explored for different clinical imaging applications. Traditional delay-and-sum (DAS) is the most common beamforming algorithm in linear array-based PA detection. Side-lobes and reconstruction-related artifacts make the DAS performance unsatisfactory and poor for a clinical-implementation. In this work, we explored a new weighting-based image processing technique for LED-based PAs to yield improved image quality when compared to the traditional methods. METHODS: We are proposing a lag-coherence factor (LCF), which is fundamentally based on the combination of the spatial auto-correlation of the detected PA signals. In LCF, the numerator contains lag-delay-multiply-and-sum (DMAS) beamformer instead of a conventional DAS beamformer. A spatial auto-correlation operation is performed between the detected US array signals before using DMAS beamformer. We evaluated the new method on both tissue-mimicking phantom (2D) and human volunteer imaging (3D) data acquired using a commercial LED-based PA imaging system. RESULTS: Our novel correlation-based weighting technique showed LED-based PA image quality improvement when it is combined with conventional DAS beamformer. Both phantom and human volunteer imaging results gave a direct confirmation that by introducing LCF, image quality was improved and this method could reduce side-lobes and artifacts when compared to the DAS and coherence-factor (CF) approaches. Signal-to-noise ratio, generalized contrast-to-noise ratio, contrast ratio and spatial resolution were evaluated and compared with conventional beamformers to assess the reconstruction performance in a quantitative way. Results show that our approach offered image quality enhancement with an average signal-to-noise ratio and spatial resolution improvement of around 20% and 25% respectively, when compared with conventional CF based DAS algorithm. CONCLUSIONS: Our results demonstrate that the proposed LCF based algorithm performs better than the conventional DAS and CF algorithms by improving signal-to-noise ratio and spatial resolution. Therefore, our new weighting technique could be a promising tool to improve the performance of LED-based PA imaging and thus accelerate its clinical translation.

High-resolution imaging of the whole eye with photoacoustic microscopy.

Observation and characterization of any changes in anatomical structures of ocular components remain as a conventional technique for diagnosis, staging, therapeutic treatments, and post-treatment monitoring of any ophthalmic disorders. The existing technologies fail to provide imaging of all of the various components of the eye simultaneously at one scanning time, i.e., one can recover vital patho-physiological information (structure and bio-molecular content) of the different ocular tissue sections only one after another. This article addresses the longstanding technological challenge by use of an emerging imaging modality [photoacoustic imaging (PAI)] in which we integrated a synthetic aperture reconstruction technique (SAFT). Experimental results-with experiments being conducted in excised tissues (goat eye)-demonstrated that we can simultaneously image the entire structure of the eye (∼2.5 cm) depicting clearly the distinctive ocular structures (cornea, aqueous humor, iris, pupil, eye lens, vitreous humor, and retina). This study uniquely opens an avenue for promising ophthalmic (clinical) applications of high clinical impact.

Energy compensated synthetic aperture focusing technique for photoacoustic microscopy.

We report an adaptive energy-compensated synthetic aperture focusing technique (eC-SAFT) for improving the imaging performance of photoacoustic microscopy (PAM) in terms of depth of field (DOF), spatial resolution (both axial and lateral), and SNR. In addition to coherency and time-delay (in conventional SAFT), our beamforming-based reconstruction algorithm takes into account acoustic energy loss-a primary physical parameter in acoustic wave propagation-following Beer-Lambert's law. Experimental validation studies were performed in tissue-mimicking (Agar) phantoms, complex leaf veins, and chicken breast tissues. Results demonstrate that our proposed eC-SAFT+CF outperforms conventional SAFT+CF to improve axial resolution (up to ∼ 5 % ), lateral resolution (up to ∼ 5 % ), SNR (up to ∼ 6 % ) and CR (up to ∼ 8 % ).

Higher-order correlation based real-time beamforming in photoacoustic imaging.

Although a delay-and-sum (DAS) beamformer is best suited for real-time photoacoustic (PA) image formation, the reconstructed images are often afflicted by noises, sidelobes, and other intense artifacts due to inaccurate assumptions in PA signal correlation. The present work aims to develop a reconstruction method that reduces the occurrence of sidelobes and artifacts and thus improves the reconstructed image quality or imaging performance. This beamformer is fundamentally based on higher-order signal correlation wherein a higher number of delayed PA signals-compared to conventional delay-multiply-and-sum (DMAS)-are combined and summed up. The proposed technique provides significant improvements in resolution, contrast, and signal-to-noise ratio (SNR) compared to traditional beamformers. For real-time implementation, the proposed algorithms were simplified, and their computational complexities were shrunk to the order of DAS [O(N)]. A GPU based study was also performed to validate the real-time capability of the proposed beamformers. For validation studies, both numerical simulation and experiments were conducted. Quantitative evaluation studies involving SNR, contrast ratio, generalized contrast-to-noise ratio, and FWHM demonstrate that the proposed higher-order DMAS beamformer is superior in PA image reconstruction. Conclusively, the proposed beamformer uniquely facilitates real-time PA image reconstruction with an achievable frame rate close to DAS and DMAS but with better imaging performance, which holds promise for real-time PA imaging and its clinical applications.

Identification, functional characterization, assembly and structure of ToxIN type III toxin-antitoxin complex from E. coli.

Toxin-antitoxin (TA) systems are proposed to play crucial roles in bacterial growth under stress conditions such as phage infection. The type III TA systems consist of a protein toxin whose activity is inhibited by a noncoding RNA antitoxin. The toxin is an endoribonuclease, while the antitoxin consists of multiple repeats of RNA. The toxin assembles with the individual antitoxin repeats into a cyclic complex in which the antitoxin forms a pseudoknot structure. While structure and functions of some type III TA systems are characterized, the complex assembly process is not well understood. Using bioinformatics analysis, we have identified type III TA systems belonging to the ToxIN family across different Escherichia coli strains and found them to be clustered into at least five distinct clusters. Furthermore, we report a 2.097 Å resolution crystal structure of the first E. coli ToxIN complex that revealed the overall assembly of the protein-RNA complex. Isothermal titration calorimetry experiments showed that toxin forms a high-affinity complex with antitoxin RNA resulting from two independent (5' and 3' sides of RNA) RNA binding sites on the protein. These results further our understanding of the assembly of type III TA complexes in bacteria.

Novel class of water-soluble phosphonate silver cluster assembled material for efficient photoelectric sensing and photoacoustic imaging.

Owing to the atomic precession and exotic photophysical properties, silver cluster assembled materials (CAMs) have been explored for use as functional nanomaterials in recent years. Although a small number of thiolate protected silver CAMs have previously been investigated, the synthesis of thiol-free analogues and their solubility remain challenging. Here, the structure-property correlation of a newly synthesized one-dimensional phenyl phosphonate protected [Ag2(PhPO3H)2(apy)2], (in which, 4,4'-azopyridine = apy) CAM is demonstrated. The multifunctional surface protecting ligand is strategically attached to the core for the first time to tailor the solubility, structural stability and charge transfer mechanism. The small size of the cluster building blocks, along with the choice of organic linker molecules, efficiently stabilize the structure via intra-chain π-π stacking while inter-chains π-π interactions create a two-dimensional supramolecular architecture. The advantageous band structure associated with the charge transfer phenomenon and the high structural stability of the material are guided to explore the sustainable photoresponsive character of this CAM, resulting in the generation of an 82 nA photocurrent. Additionally, the unprecedented water solubility, which is very rare for this class of material, provides opportunities for use in biomedical imaging applications. The measured photoacoustic signal strength confirms the blood vessel mimicking capabilities of the portrayed material at a depth of approximately 3 mm inside chicken breast tissue.

Delay-and-sum-to-delay-standard-deviation factor: a promising adaptive beamformer.

A new adaptive weighting method [delay-and-sum-to-delay-standard-deviation factor (DASDSF)] combined with minimum variance (MV) beamforming is introduced in photoacoustic imaging (PAI). Existing MV-based beamformers improve photoacoustic image quality in terms of achieving narrow main lobes and, thus, improving spatial resolution. But, the beamformers give a strong side-lobe signal strength that greatly degrades the reconstructed image contrast. As a feedback weighting factor, DASDSF addresses the persisting side-lobe issue present in MV-beamformed images, i.e., our proposed method is robust against reduction in noises as well as side lobes, and it outperforms MV and MV combined with coherence factor beamformers. Validation studies-being carried out both in numerical simulation and experiments employing a low-cost (16 elements) linear transducer array in a home-built PAI system-demonstrate an excellent performance of the proposed weighting approach in improving SNR, while reducing main-lobe width (i.e., FWHM) and side-lobe signal strength. The study demonstrates that the proposed algorithm holds promise for development of a cost-effective PAI system using a low-cost linear transducer (∼16 elements against ∼128 generally used).

Enhancement of Photoacoustic Signal Strength with Continuous Wave Optical Pre-Illumination: A Non-Invasive Technique.

Use of portable and affordable pulse light sources (light emitting diodes (LED) and laser diodes) for tissue illumination offers an opportunity to accelerate the clinical translation of photoacoustic imaging (PAI) technology. However, imaging depth in this case is limited because of low output (optical) power of these light sources. In this work, we developed a noninvasive technique for enhancing strength (amplitude) of photoacoustic (PA) signal. This is a photothermal-based technique in which a continuous wave (CW) optical beam, in addition to short-pulse ~ nsec laser beam, is employed to irradiate and, thus, raise the temperature of sample material selectively over a pre-specified region of interest (we call the process as pre-illumination). The increase in temperature, in turn enhances the PA-signal strength. Experiments were conducted in methylene blue, which is one of the commonly used contrast agents in laboratory research studies, to validate change in temperature and subsequent enhancement of PA-signal strength for the following cases: (1) concentration or optical absorption coefficient of sample, (2) optical power of CW-optical beam, and (3) time duration of pre-illumination. A theoretical hypothesis, being validated by numerical simulation, is presented. To validate the proposed technique for clinical and/or pre-clinical applications (diagnosis and treatments of cancer, pressure ulcers, and minimally invasive procedures including vascular access and fetal surgery), experiments were conducted in tissue-mimicking Agar phantom and ex-vivo animal tissue (chicken breast). Results demonstrate that pre-illumination significantly enhances PA-signal strength (up to ~70% (methylene blue), ~48% (Agar phantom), and ~40% (chicken tissue)). The proposed technique addresses one of the primary challenges in the clinical translation of LED-based PAI systems (more specifically, to obtain a detectable PA-signal from deep-seated tissue targets).