Powdered plant beverages obtained by spray-drying without carrier addition-physicochemical and chemometric studies.

Plant-based beverages (PBs) are currently gaining interest among consumers who are seeking alternative sustainable options to traditional dairy drinks. The study aimed to obtain powdered plant beverages without the addition of carriers by spray drying method to implement them in the future as an alternative to the liquid form of dairy drinks. Some of the most well-known commercial beverages sources like soy, almond, rice and oat were analyzed in this work. The effect of different treatments (concentration, addition of oat fiber) and two approaches od spray drying (conventional high temperature spray drying-SD, and dehumidified air spray drying at low temperature-DASD) were presented. Moreover, moisture content, water activity, particle morphology and size of obtained powders were analyzed. It was possible to obtain PBs without the addition of carriers, although the drying yield of four basic beverages was low (16.1-37.4%). The treatments and change in spray drying approach enhanced the drying yield, especially for the concentrated beverage dried using DASD (59.2%). Additionally, Fourier Transform Infrared (FTIR) spectroscopy was applied to evaluate the differences in chemical composition of powdered PBs. FTIR analysis revealed differences in the range of the absorption frequency of amide I, amide II (1700-1500 cm-1) and carbohydrate region (1200-900 cm-1). Principal component analysis (PCA) was carried out to study the relationship between spray dried plant beverages samples based on the fingerprint region of FTIR spectra, as well as the physical characteristics. Additionally, hierarchical cluster analysis (HCA) was employed to explore the clustering of the powders.

Diallyl disulfide induces DNA damage and growth inhibition in colorectal cancer cells by promoting POU2F1 ubiquitination.

Previous studies have demonstrated that diallyl disulfide (DADS) exhibits potent anti-tumor activity. However, the pharmacological actions of DADS in inhibiting the growth of colorectal cancer (CRC) cells have not been clarified. Herein, we show that DADS treatment impairs the activation of the pentose phosphate pathway (PPP) to decrease PRPP (5-phosphate ribose-1-pyrophosphate) production, enhancing DNA damage and cell apoptosis, and inhibiting the growth of CRC cells. Mechanistically, DADS treatment promoted POU2F1 K48-linked ubiquitination and degradation by attenuating the PI3K/AKT signaling to up-regulate TRIM21 expression in CRC cells. Evidently, TRIM21 interacted with POU2F1, and induced the K272 ubiquitination of POU2F1. The effects of DADS on the enhanced K272 ubiquitination of POU2F1, the PPP flux, PRPP production, DNA damage and cell apoptosis as well as the growth of CRC tumors in vivo were significantly mitigated by TRIM21 silencing or activating the PI3K signaling in CRC cells. Conversely, the effects of DADS were enhanced by TRIM21 over-expression or inhibiting the PI3K/AKT signaling in CRC cells. Collectively, our findings reveal a novel mechanism by which DADS suppresses the growth of CRC by promoting POU2F1 ubiquitination, and may aid in design of novel therapeutic intervention of CRC.

Upper and lower limb tremor in inflammatory neuropathies.

OBJECTIVE: The mechanisms underlying neuropathic tremor remain incompletely understood and a distinction has not been drawn between proximal and distal neuropathies. Lower limb tremor contributes to imbalance in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), but this is unexplored in other neuropathies. We characterized upper and lower limb tremor in chronic immune sensory polyradiculopathy (CISP) and distal acquired demyelinating neuropathy with anti-MAG antibodies (DADS-MAG), contrasted to CIDP. METHODS: This was a cross-sectional study of 38 patients (CIDP [n = 25], CISP [n = 7], DADS-MAG [n = 6]). Clinical assessment, tremor study recordings, nerve conduction studies, and somatosensory evoked potentials were performed. Balance was measured by force platform. RESULTS: Upper limb tremor was prevalent (CIDP 66%, CISP 70%, DADS-MAG 100%). Peak frequencies followed a gradient along the upper limb, unchanged by weight-loading. Lower limb tremor was also present (CIDP 32%, CISP 29%, DADS-MAG 66%) and associated with imbalance. Nerve conduction parameters correlated with upper limb tremor in DADS-MAG and CISP, and imbalance in CISP. CONCLUSIONS: Upper limb tremor is mediated by peripheral and central mechanisms regardless of distal or proximal pathology. Lower limb tremor correlates with peripheral nerve function and contributes to imbalance. SIGNIFICANCE: This study contributes to the understanding of neuropathic tremor. Addressing lower limb tremor may be of therapeutic importance for neuropathy-associated imbalance.

DADS Inhibits the Proliferation of MDCC-MSB-1 Cells by Inducing Autophagy via the MEK/ERK Signalling Pathway.

Diallyl disulfide (DADS) is effective at suppressing tumour cell growth and proliferation. This study verified the morphology and growth activity of MDCC-MSB-1 cells by using an MTT assay to detect the effect of DADS on the proliferation of MDCC-MSB-1 cells and a CCK8 assay to detect the effect of DADS on the viability and proliferation of MDCC-MSB-1 cells. We found that the viability and proliferation of MDCC-MSB-1 cells decreased with increasing DADS concentrations. MDC staining and Western blotting were used to analyse autophagy, the associated protein LC3 and the MEK/ERK pathway proteins MEK and ERK and to investigate changes in cellular autophagy based on cell morphology and molecular biology. With increasing concentrations of DADS, MDCC-MSB-1 cell autophagy increased in a gradient manner. Additionally, the conversion of the autophagy marker protein LC3-I increased with increasing drug concentrations, and the relative expression of LC3-II steadily increased, as did the expression of key protein components of the MEK/ERK signalling pathway, including P-MEK1/2 and P-ERK1/2. These results suggest that DADS induces autophagy through the MEK/ERK pathway, thereby inhibiting the proliferation of MDCC-MSB-1 cells.

Diallyl disulfide synergizes with melphalan to increase apoptosis and DNA damage through elevation of reactive oxygen species in multiple myeloma cells.

Diallyl disulfide (DADS), one of the main components of garlic, is well known to have anticancer effects on multiple cancers. However, its efficacy in treating multiple myeloma (MM) is yet to be determined. We explored the effects of DADS on MM cells and investigated the synergistic effects of DADS when combined with five anti-MM drugs, including melphalan, bortezomib, carfilzomib, doxorubicin, and lenalidomide. We analyzed cell viability, cell apoptosis, and DNA damage to determine the efficacy of DADS and the drug combinations. Our findings revealed that DADS induces apoptosis in MM cells through the mitochondria-dependent pathway and increases the levels of γ-H2AX, a DNA damage marker. Combination index (CI) measurements indicated that the combination of DADS with melphalan has a significant synergistic effect on MM cells. This was further confirmed by the increases in apoptotic cells and DNA damage in MM cells treated with the two drug combinations compared with those cells treated with a single drug alone. The synergy between DADS and melphalan was also observed in primary MM cells. Furthermore, mechanistic investigations showed that DADS decreases reduced glutathione (GSH) levels and increases reactive oxygen species (ROS) production in MM cells. The addition of GSH is effective in neutralizing DADS cytotoxicity and inhibiting the synergy between DADS and melphalan in MM cells. Taken together, our study highlights the effectiveness of DADS in treating MM cells and the promising therapeutic potential of combining DADS and melphalan for MM treatment.

The protein architecture and allosteric landscape of HNF4α.

Hepatocyte nuclear factor 4 alpha (HNF4α) is a multi-faceted nuclear receptor responsible for governing the development and proper functioning of liver and pancreatic islet cells. Its transcriptional functions encompass the regulation of vital metabolic processes including cholesterol and fatty acid metabolism, and glucose sensing and control. Various genetic mutations and alterations in HNF4α are associated with diabetes, metabolic disorders, and cancers. From a structural perspective, HNF4α is one of the most comprehensively understood nuclear receptors due to its crystallographically observed architecture revealing interconnected DNA binding domains (DBDs) and ligand binding domains (LBDs). This review discusses key properties of HNF4α, including its mode of homodimerization, its binding to fatty acid ligands, the importance of post-translational modifications, and the mechanistic basis for allosteric functions. The surfaces linking HNF4α's DBDs and LBDs create a convergence zone that allows signals originating from any one domain to influence distant domains. The HNF4α-DNA complex serves as a prime illustration of how nuclear receptors utilize individual domains for specific functions, while also integrating these domains to create cohesive higher-order architectures that allow signal responsive functions.

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.

Induce defense response of DADS in eggplants during the biotrophic phase of Verticillium dahliae.

PURPOSE: Verticillium wilt is a destructive vascular disease in eggplants. The complex defensive mechanisms of eggplant against this disease are very limited. METHODS: Our work examined the bioactive properties of garlic allelochemical diallyl disulfide (DADS) as potential biostimulants for defense against V. dahliae in eggplant seedlings. We, therefore, foliar sprayed DADS on eggplants to study the defense response during the early biotrophic phase of V. dahliae (a hemibiotroph). RESULTS: DADS application significantly increased root peroxidase (POD), phenylalanine-ammonia lyase (PAL) enzyme activity, and reduced H2O2 levels after 24 h of fungal inoculation. Salicylic acid (SA) in leaves and roots was significantly increased while, the jasmonic acid (JA), indole acetic acid (IAA), and abscisic acid (ABA) levels were decreased. The microscopic examinations of V. dahliae infection in roots displayed that the progression of infection was restricted in DADS-treated plants. Depositions of lignin and phenolic compounds such as ferulic acid, p-coumaric acid, and caffeic acid content were significantly higher in DADS-treated plants at 48 h post-inoculation. Similarly, the DADS application up-regulated pathogenesis-related (PR1, PR2, and PR5), mitogen-activated protein kinase (MPK1), and lipoxygenase (LOX) genes. Furthermore, DADS-treated plants exhibited a lower disease severity index (23.3% vs. 57.0% in controls), indicating successful defense against V. dahliae. CONCLUSIONS: Our findings concluded that the biological function of garlic allelochemical DADS has a prominent role in the higher defense resistance of eggplants during the early infection of V. dahliae.

DTU-DADS-Aqua: A simulation framework for modelling waterborne spread of highly infectious pathogens in marine aquaculture.

Simulation models are useful tools to predict and elucidate the effects of factors influencing the occurrence and spread of epidemics in animal populations, evaluate the effectiveness of different control strategies and ultimately inform decision-makers about mitigations to reduce risk. There is a paucity of simulation models to study waterborne transmission of viral and bacterial pathogens in marine environments. We developed a stochastic, spatiotemporal hybrid simulation model (DTU-DADS-Aqua) that incorporates a compartmental model for infection spread within net-pens, an agent-based model for infection spread between net-pens within and between sites and uses seaway distance to inform farm-site hydroconnectivity. The model includes processes to simulate infection transmission and control over surveillance, detection and depopulation measures. Different what-if scenarios can be explored according to the input data provided and user-defined parameter values, such as daily surveillance and depopulation capacities or increased animal mortality that triggers diagnostic testing to detect infection. The latter can be easily defined in a software application, in which results are summarized after each simulation. To demonstrate capabilities of the model, we simulated the spread of infectious salmon anaemia virus (ISAv) for realistic scenarios in a transboundary population of farmed Atlantic salmon (Salmo salar L.) in New Brunswick, Canada and Maine, United States. We assessed the progression of infection in the different simulated outbreak scenarios, allowing for variation in the control strategies adopted for ISAv. Model results showed that improved disease detection, coupled with increasing surveillance visits to farm-sites and increased culling capacity for depopulation of infected net-pens reduced the number of infected net-pens and outbreak duration but the number of ISA-infected farm sites was minimally affected. DTU-DADS-Aqua is a flexible modelling framework, which can be applied to study different infectious diseases in the aquatic environment, allowing the incorporation of alternative transmission and control dynamics. The framework is open-source and available at https://github.com/upei-aqua/DTU-DADS-Aqua.

Molecular Determinant of DIDS Analogs Targeting RAD51 Activity.

RAD51 is the central protein in DNA repair by homologous recombination (HR), involved in several steps of this process. It is shown that overexpression of the RAD51 protein is correlated with increased survival of cancer cells to cancer treatments. For the past decade, RAD51 overexpression-mediated resistance has justified the development of targeted inhibitors. One of the first molecules described to inhibit RAD51 was the 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS) molecule. This small molecule is effective in inhibiting different functions of RAD51, however its mode of action and the chemical functions involved in this inhibition have not been identified. In this work, we used several commercial molecules derived from DIDS to characterize the structural determinants involved in modulating the activity of RAD51. By combining biochemical and biophysical approaches, we have shown that DIDS and two analogs were able to inhibit the binding of RAD51 to ssDNA and prevent the formation of D-loop by RAD51. Both isothiocyanate substituents of DIDS appear to be essential in the inhibition of RAD51. These results open the way to the synthesis of new molecules derived from DIDS that should be greater modulators of RAD51 and more efficient for HR inhibition.

Field efficacy of Bt cotton containing events DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 against lepidopteran pests and impact on the non-target arthropod community in Brazil.

The efficacy and non-target arthropod effects of transgenic DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 Bt cotton, expressing proteins Cry1Ac, Cry1F and Vip3Aa19, was examined through field trials in Brazil. Fifteen field efficacy experiments were conducted from 2014 through the 2020 growing season across six different states in Brazil to evaluate performance against key lepidopteran pests through artificial infestations of Chrysodeixis includens (Walker), Spodoptera frugiperda (J.E. Smith,1797), Spodoptera cosmioides (Walker, 1858) and Chloridea virescens (F., 1781), and natural infestations of Alabama argillacea (Hübner) and S. frugiperda. The impact of this Bt cotton technology on the non-target arthropod community in Brazilian cotton production systems was also assessed in a multi-site experiment. DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton significantly reduced the feeding damage caused by S. frugiperda, S. cosmioides, C. includens, C. virescens and A. argillacea, causing high levels of mortality (greater than 99%) to all target lepidopteran pests evaluated during vegetative and/or reproductive stages of crop development. Non-target arthropod community-level analyses confirmed no unintended effects on the arthropod groups monitored. These results demonstrate the value of transgenic Bt cotton containing event DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 for consideration as part of an integrated approach for managing key lepidopteran pests in Brazilian cotton production systems.

The role of HCO3- in propionate-induced anion secretion across rat caecal epithelium.

Propionate, a metabolite from the microbial fermentation of carbohydrates, evokes a release of epithelial acetylcholine in rat caecum resulting in an increase of short-circuit current (Isc) in Ussing chamber experiments. The present experiments were performed in order to characterize the ionic mechanisms underlying this response which has been thought to be due to Cl- secretion. As there are regional differences within the caecal epithelium, the experiments were conducted at oral and aboral rat corpus caeci. In both caecal segments, the propionate-induced Isc (IProp) was inhibited by > 85%, when the experiments were performed either in nominally Cl-- or nominally HCO3--free buffer. In the case of Cl-, the dependency was restricted to the presence of Cl- in the serosal bath. Bumetanide, a blocker of the Na+-K+-2Cl--cotransporter, only numerically reduced IProp suggesting that a large part of this current must be carried by an ion other than Cl-. In the aboral caecum, IProp was significantly inhibited by mucosally administered stilbene derivatives (SITS, DIDS, DNDS), which block anion exchangers. Serosal Na+-free buffer reduced IProp significantly in the oral (and numerically also in aboral) corpus caeci. RT-PCR experiments revealed the expression of several forms of Na+-dependent HCO3--cotransporters in caecum, which might underlie the observed Na+ dependency. These results suggest that propionate sensing in caecum is coupled to HCO3- secretion, which functionally would stabilize luminal pH when the microbial fermentation leads to an increase in the concentration of short-chain fatty acids in the caecal lumen.

Several phosphate transport processes are present in vascular smooth muscle cells.

We have studied inorganic phosphate (Pi) handling in rat aortic vascular smooth muscle cells (VSMC) using 32P-radiotracer assays. Our results have revealed a complex set of mechanisms consisting of 1) well-known PiT1/PiT2-mediated sodium-dependent Pi transport; 2) Slc20-unrelated sodium-dependent Pi transport that is sensitive to the stilbene derivatives 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) and 4-acetamido-4-isothiocyanostilbene-2,2-disulfonate (SITS); 3) a sodium-independent Pi uptake system that is competitively inhibited by sulfate, bicarbonate, and arsenate and is weakly inhibited by DIDS, SITS, and phosphonoformate; and 4) an exit pathway from the cell that is partially chloride dependent and unrelated to the known anion-exchangers expressed in VSMC. The inhibitions of sodium-independent Pi transport by sulfate and of sodium-dependent transport by SITS were studied in greater detail. The maximal inhibition by sulfate was similar to that of Pi itself, with a very high inhibition constant (212 mM). SITS only partially inhibited sodium-dependent Pi transport, but the Ki was very low (14 µM). Nevertheless, SITS and DIDS did not inhibit Pi transport in Xenopus laevis oocytes expressing PiT1 or PiT2. Both the sodium-dependent and sodium-independent transport systems were highly dependent on VSMC confluence and on the differentiation state, but they were not modified by incubating VSMC for 7 days with 2 mM Pi under nonprecipitating conditions. This work not only shows that the Pi handling by cells is highly complex but also that the transport systems are shared with other ions such as bicarbonate or sulfate.NEW & NOTEWORTHY In addition to the inorganic phosphate (Pi) transporters PiT1 and PiT2, rat vascular smooth muscle cells show a sodium-dependent Pi transport system that is inhibited by DIDS and SITS. A sodium-independent Pi uptake system of high affinity is also expressed, which is inhibited by sulfate, bicarbonate, and arsenate. The exit of excess Pi is through an exchange with extracellular chloride. Whereas the metabolic effects of the inhibitors, if any, cannot be discarded, kinetic analysis during initial velocity suggests competitive inhibition.

Safety evaluation of DAS-44406-6 soybeans in Wistar rats.

A 90-day in-country feeding trial in Wistar rats was conducted at Tianjin Laboratory in China to assess toxicity of diets containing DAS-44406-6 soybean meal. There were no treatment-related changes observed when compared with the non-GM isoline control groups but histopathologically, 2 of 10 high-dose females were reported to show kidney lesions. However, these findings contrasted with the absence of any treatment-related kidney lesions in 3 separate 90-day toxicity studies previously conducted in Sprague Dawley rats. Strain difference is not expected in the kidney response, and based on the low incidence and contrary evidence from previous studies, it is likely that these lesions were of spontaneous origin, or artefactual. To determine that the lesions observed were not treatment-related in Wistar rats, a specific follow-up confirmatory study was conducted under Good Laboratory Practices (GLP) in the Wistar strain of rats following an identical study design to the Tianjin study. To increase the power of detecting effects, twice the number of animals per group (20/sex/group) were used, and no treatment-related kidney histopathological changes were observed. Based on these results and entire weight of evidence evaluation, it is concluded that the histopathological changes previously noted in the 2 female Wistar rats of Tianjin study were not treatment-related and that DAS-44406-6 soybeans are as safe as conventional non-GM soybeans.

Sustained-released mixture of vascular endothelial growth factor 165 and fibrin glue strengthens healing of ileal anastomoses in a rabbit model with intraperitoneal infection

PURPOSE: To investigate the effects of a sustained-released mixture of vascular endothelial growth factor 165 (VEGF165) and fibrin glue (FG) local administration on postoperative rabbit ileal anastomoses. METHODS: One hundred twenty-eight male and female New Zealand white rabbits underwent intraperitoneal infection subsequent ileal anastomosis surgery were divided randomly into 4 groups, including 32 animals in each, applied with saline solution, FG, rhVEGF165 and a mixture of rhVEGF165 with FG (VEGF + FG) on the anastomoses, respectively. The incidences of anastomotic leakage were observed. Histopathological examination for inflammatory infiltration, fibroblast proliferation, and capillary vascular proliferation were performed. Then, bursting pressure and hydroxyproline concentrations were assessed in anastomoses sits on postoperative days 3, 5, 7, and 14. RESULTS: Rabbits in VEGF + FG group had the lowest incidence of leakage (P < 0.05). Histological evaluations revealed that granulation tissue was formed on days 5 after anastomosis; fibroblast proliferation and capillary vascular proliferation were significantly increased on days 7 and 14 in VEGF + FG group. Furthermore, there was a statistically significant difference in the mean bursting pressures between VEGF + FG group and other groups on days 7 and 14 (P < 0.05), and rabbits in VEGF + FG group exhibited a higher concentration than VEGF group (P < 0.05) and FG group (P < 0.05) on day 14. CONCLUSION: Administration of VEGF165 mixed with FG to ileal anastomosis accelerates wound healing and enhances the anastomosis by increased angiogenesis.

Event DAS-444Ø6-6 soybean grown in Brazil is compositionally equivalent to non-transgenic soybean.

Soybean event DAS-444Ø6-6 is tolerant to the herbicides 2,4-D, glyphosate, and glufosinate. An investigation of potential unintended adverse compositional changes in a genetically modified crop is required to meet government regulatory requirements in various geographies. A study to meet these requirements in Brazil was completed demonstrating compositional equivalency between DAS-444Ø6-6 and non-transgenic soybean. This study supplements the extensive literature supporting transgenesis as less disruptive of crop composition compared with traditional breeding methods.

Current Status and Challenges of Cancer Clinical Trials in Korea / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Cancer clinical trials in Korea have rapidly progressed in terms of quantity and quality during the last decade. This study evaluates the current status of cancer clinical trials in Korea and their associated problems. MATERIALS AND METHODS: We analyzed the clinical trials approved by the Korea Food and Drug Administration (KFDA) between 2007 and 2013. A nationwide on-line survey containing 22 questions was also performed with several cooperative study groups and individual researchers in 56 academic hospitals. RESULTS: The number of cancer clinical trials approved by the KFDA increased almost twofold from 2007 to 2013. The number of sponsor-initiated clinical trials (SITs) increased by 50% and investigator-initiated clinical trials (IITs) increased by almost 640%. Three hundred and forty-four clinical trials were approved by the KFDA between 2012 and 2013. At the time of the on-line survey (August 2013), 646 SITs and 519 IITs were ongoing in all hospitals. Six high volume hospitals were each conducting more than 50 clinical trials, including both SITs and IITs. Fifty-six investigators (31%) complained of the difficulties in raising funds to conduct clinical trials. CONCLUSION: The number of cancer clinical trials in Korea rapidly increased from 2007 to 2013, as has the number of multicenter clinical trials and IITs run by cooperative study groups. Limited funding for IIT is a serious problem, and more financial support is needed both from government agencies and public donations from non-profit organizations.