Anti-Warburg effect via generation of ROS and inhibition of PKM2/ß-catenin mediates apoptosis of lambertianic acid in prostate cancer cells.

To elucidate the underlying antitumor mechanism of lambertianic acid (LA) derived from Pinus koraiensis, the role of cancer metabolism related molecules was investigated in the apoptotic effect of LA in DU145 and PC3 prostate cancer cells. MTT assay for cytotoxicity, RNA interference, cell cycle analysis for sub G1 population, nuclear and cytoplasmic extraction, lactate, Glucose and ATP assay by ELISA, Measurement of reactive oxygen species (ROS) generation, Western blotting, and immunoprecipitation assay were conducted in DU145 and PC3 prostate cancer cells. Herein LA exerted cytotoxicity, increased sub G1 population and attenuated the expression of pro-Caspase3 and pro-poly (ADP-ribose) polymerase (pro-PARP) in DU145 and PC3 cells. Also, LA reduced the expression of lactate dehydrogenase A (LDHA), glycolytic enzymes such as hexokinase 2 and pyruvate kinase M2 (PKM2) with reduced production of lactate in DU145 and PC3 cells. Notably, LA decreased phosphorylation of PKM2 on Tyr105 and inhibited the expression of p-STAT3, cyclin D1, C-Myc, ß-catenin, and p-GSK3ß with the decrease of nuclear translocation of p-PKM2. Furthermore, LA disturbed the binding of p-PKM2 and ß-catenin in DU145 cells, which was supported by Spearman coefficient (0.0463) of cBioportal database. Furthermore, LA generated ROS in DU145 and PC3 cells, while ROS scavenger NAC (N-acetyl L-cysteine) blocked the ability of LA to reduce p-PKM2, PKM2, ß-catenin, LDHA, and pro-caspase3 in DU145 cells. Taken together, these findings provide evidence that LA induces apoptosis via ROS generation and inhibition of PKM2/ß-catenin signaling in prostate cancer cells.

Improvement of pasting and gelling properties of potato starch using a direct vapor-heat moisture treatment.

Conventional hydrothermal methods are lengthy and require high energy consumption. In this study, a quick and energy-saving direct vapor-heat moisture treatment (DV-HMT) method was used to improve the pasting and gelling properties of potato starch under the condition of high temperature (130 °C) and short periods (1, 3, 5, and 7 min). X-ray diffraction analysis exhibited that the relative crystallinity of DV-HMT starches decreased with the extension of treatment time. Small angle X-ray scattering measurements showed that the average thickness of the crystalline lamellae decreased from 6.193 to 5.937 nm, while the average thickness of the amorphous lamellae increased from 3.160 to 3.395 nm. Rapid visco-analyzer measurements exhibited that the breakdown values decreased to 0 mPa s for DV-HMT starches, indicating that this hydrothermal treatment led to starches with high resistance to heating and shearing. The gel hardness of starch treated by DV-HMT for 3 min (266 g) was around 5.4-fold higher than for non-treated starch (41 g). Considering the simple operation of DV-HMT and the short treatment periods (≤ 7 min) used in this study, DV-HMT could be a superior option to enhance the physicochemical and functional properties of starch.

Engineered Organosilica Hybrid Micelles for Photothermal-enhanced Starvation Cancer Therapy.

Glucose oxidase (GOD)-based starvation therapy (ST), which inhibits the growth and proliferation of cancer cells by consuming glucose, has attracted intensive attention as an emerging non-invasive method for fighting cancers. However, the enzyme activity of GOD is greatly limited in vivo because of its optimal catalytic activity in the temperature range of 43-60 °C. Herein, a photothermal-enhanced starvation strategy is developed based on our engineered organosilica hybrid micelles (TiO2-x @POMs-GOD), in which the fluoride-doped TiO2-x with photothermal properties is encapsulated in the cores of organosilica cross-linked micelles and GOD is immobilized on the carboxyl groups of PAA segments. With its internalization by cancer cells, the conjugated GOD can effectively deplete glucose to achieve the ST effect, which can be remarkably enhanced by the loaded fluoride-doped TiO2-x with NIR laser irradiation, thus cooperatively contributing to the efficient treatment of TiO2-x @POMs-GOD on various cancer cells. This suggests great potential for TiO2-x @POMs-GOD in photothermal-enhanced ST in vivo.

Antitumor mechanism of combination of Angelica gigas and Torilis japonica in LNCaP prostate cancer cells via G1 arrest and inhibition of Wnt/ß-catenin and androgen receptor signaling.

The goal of the current study is to assess the antitumor mechanism by the combination (7:3) of Angelica gigas and Torilis japonica (AT) that was found most effective through screening against prostate-specific antigen (PSA) in LNCaP prostate cancer cells. Here, AT reduced the viability and the number of colonies in androgen-dependent LNCaP cells more than in androgen independent PC3 and DU145 cells. Also, AT induced G1 phase arrest, cleaved PARP and caspase 3, activated p27 and decreased the expression of Cyclin D1, Cyclin E, cdk2 in LNCaP cells. Furthermore, AT decreased the expression of PSA and androgen receptor (AR) at mRNA and protein levels in LNCaP cells. Interestingly, AT attenuated the expression of AR, PSA and Wnt-3a and the stability of AR and PSA in LNCaP cells. Furthermore, AT reversed dihydrotestosterone (DHT)-induced upregulation of AR and PSA in LnCaP cells. Notably, AT disrupted the protein-protein interaction, nuclear translocation and fluorescent expression of ß-catenin and AR in LNCaP cells. Consistently, ß-catenin depletion enhanced the decreased expression of AR in AT treated LNCaP cells. Taken together, our findings highlight evidence that AT suppresses the proliferation of LNCaP cells via G1 arrest and inhibition of ß-catenin and AR as a potential anticancer agent.

Comparison of Lutein Bioaccessibility from Dietary Supplement-Excipient Nanoemulsions and Nanoemulsion-Based Delivery Systems.

The impact of lutein-loaded nanoemulsions and excipient nanoemulsions mixed with lutein-based dietary supplements (capsules and soft gels) on the bioaccessibility of lutein was explored using a simulated gastrointestinal tract (GIT). The particle size, particle size distribution, ζ-potential, microstructure, lipid digestibility, and lutein bioaccessibility of all the samples were measured after they were exposed to different environments (stomach and small intestine environments) within a simulated GIT. As expected, the bioaccessibility of lutein from the capsules (1.5%) and soft gels (3.2%) was relatively low when they were administered alone. However, the co-administration of excipient nanoemulsions significantly increased the bioaccessibility of lutein from both the capsules (35.2%) and soft gels (28.7%). This phenomenon was attributed to the fast digestion of the small oil droplets in the excipient nanoemulsions and the further formation of mixed micelles to solubilize any lutein molecules released from the supplements. The lutein-loaded nanoemulsions exhibited a much higher lutein bioaccessibility (86.8%) than any of the supplements, which was attributed to the rapid release and solubilization of lutein when the lipid droplets were rapidly and extensively digested within the small intestine. This study indicates that the bioaccessibility of lutein is much higher in nanoemulsion droplets than that in dietary supplements. However, consuming dietary supplements in the presence of nanoemulsion droplets can greatly increase lutein bioavailability. The results of this study have important guiding significance for the design of more effective lutein supplements.

Anti-freezing starch hydrogels with superior mechanical properties and water retention ability for 3D printing.

As a novel material that can be used at subzero temperatures, anti-freezing hydrogels have been attracting extensive attention. Inspired by the freeze-tolerance phenomenon in seawater, which is achieved by mixing salts into water, an ionic compound (CaCl2) was used to gelatinize starch to form anti-freezing hydrogels. Native potato starch (NPS) anti-freezing hydrogels were formed at -10 °C, -18 °C, -30 °C, and - 50 °C with 6-9 kPa tensile strength and 100-230% elongation at break. The compressive stress of anti-freezing hydrogels at different environmental temperatures increased from 18.586 kPa to 36.551 kPa with the glass transform temperature of starch hydrogels dropped to -50 °C. The anti-freezing hydrogels showed excellent water retention ability, which could maintain a water content of 55% after 7 days at ambient temperature. The prototyping of anti-freezing starch hydrogels broadens the applications of starch in food, adhesives, medical materials, and intelligent materials.

Inhibition of TMPRSS4 mediated epithelial-mesenchymal transition is critically involved in antimetastatic effect of melatonin in colorectal cancers.

In the current study, the underlying anti-metastatic mechanism of melatonin contained in some edible plants was explored in association with transmembrane protease serine 4 (TMPRSS4) mediated metastasis and epithelial-mesenchymal transition (EMT) signaling in human HCT15 and SW620 colorectal cancer cells. Here, TMPRSS4 was highly expressed in HCT15, but was weakly expressed in SW620 cells. Melatonin exerted weak cytotoxicity, decreased invasion, adhesion, and migration, and attenuated the expression of TMPRSS4, cyclin E, pro-urokinase-type plasminogen activator (pro-uPA), p-signal transducer and activator of transcription 3 (p-STAT3), p-focal adhesion kinase (p-FAK), Snail and increased the expression of E-cadherin, p27, pp38 and p-Jun N-terminal kinases (p-JNK) in HCT15 cells. Conversely, overexpression of TMPRSS4 reduced the ability of melatonin to activate E-cadherin and reduce Snail. Furthermore, even in SW620 cells transfected with TMPRSS4-overexpression plasmid, melatonin effectively suppressed invasion and migration along with decreased expression of Snail, cyclin A, cyclin E, pro-uPA and p-FAK and increased expression of E-cadherin and p27. Overall, these findings provide evidence that melatonin suppresses metastasis in colon cancer cells via inhibition of TMPRSS4 mediated EMT.

Long-Term Tea Consumption Is Associated with Reduced Risk of Diabetic Retinopathy: A Cross-Sectional Survey among Elderly Chinese from Rural Communities.

AIM: To investigate the association between variables related to tea consumption (duration, frequency, and type) and the risk of diabetic retinopathy. METHODS: A rural community-based, cross-sectional survey was conducted in Weitang Town, Suzhou, China. People aged 60 years or above were invited to complete the survey. All eligible patients underwent detailed eye examination. Diabetic retinopathy (DR) was diagnosed and graded based on the retinal fundus imaging. Diabetes was defined as fasting glucose concentrations of ≥7.0 mmol/L or self-reported diagnosis of diabetes. Information about tea consumption such as duration, type, and frequency, together with demographics and lifestyle characteristics, were collected using a face-to-face questionnaire interview. The association between tea consumption and the risk of DR was determined by univariate and multivariate logistic regression analyses. RESULTS: Among the 5,281 participants, 614 had diabetes mellitus (prevalence of 11.63%). The prevalence rate of DR was 10.38% in the diabetic population and 1.04% in the general population. Compared with non-tea consumers, the crude OR values for DR in subjects with long-term and short-term tea consumption were 0.34 (95%CI = 0.14-0.82, p = 0.016) and 1.64 (95%CI = 0.74-3.64, p = 0.221), respectively. When adjusted for age, gender, and other confounders, consumption of tea for ≥20 years was associated with reduced odds of DR (OR = 0.29, 95%CI = 0.09-0.97, p = 0.044). Thus, long-term tea consumption was significantly associated with a lower risk of DR. There was no statistical significance between frequency or type of tea consumption with DR (p > 0.05). CONCLUSION: Elderly diabetic Chinese residents who consumed tea for more than twenty years had a lower risk of DR compared to non-tea consumers. The long-term tea consumption may be an independent protective factor for DR. However, further studies are warranted to examine the association.

p53 dependent LGR5 inhibition and caspase 3 activation are critically involved in apoptotic effect of compound K and its combination therapy potential in HCT116 cells.

Though ginsenoside metabolite compound K was known to have antitumor effect in several cancers, its underlying apoptotic mechanism still remains unclear so far. Thus, in the present study, the apoptotic mechanism of compound K was explored in colorectal cancer cells (CRCs) in association with leucine rich repeat containing G protein-coupled receptor 5 (LGR5) that was overexpressed in colorectal cancers with poor survival rate. Here compound K significantly reduced viability of HCT116p53+/+ cells better than that of HCT116p53-/- cells. Consistently, compound K increased sub G1 population and attenuated the expression of LGR5, c-Myc, procaspase3, Pin1 in HCT116p53+/+ cells more than in HCT116p53-/- cells. Conversely, caspase 3 inhibitor Z-DEVD-FMK reversed inhibitory effect of compound K on LGR5, c-Myc and procaspase3 in HCT116 cells. Consistently, inhibition of LGR5 using transfection method enhanced suppression of pro-PARP, Bcl-xL c-Myc, Snail and Pin1 in compound K treated HCT116p53+/+ cells. Furthermore, compound K synergistically potentiated antitumor effect of 5-fluorouracil (5-FU) or Doxorubicin to reduce the survival genes and cytotoxicity in HCT116p53+/+ cells. Overall, our findings provide scientific insight that compound K induces apoptosis in colon cancer cells via caspase and p53 dependent LGR5 inhibition with combination therapy potential with 5-FU or doxorubicin.

LGR4 overexpression is associated with clinical parameters and poor prognosis of serous ovarian cancer.

OBJECTIVE: LGR4 expression in serous ovarian cancer paraffin-embedded tissues and fresh tissues were investigated, and its expression associated with clinicopathological parameters and prognosis in serous ovarian cancer was explored. METHODS: From Dec, 2009 to Jan, 2020, 122 paraffin-embedded serous ovarian cancer patients and 41 paired paratumor tissues who were both diagnosed and operated at the memorial hospital of Sun Yat-sen University and Integrated Hospital of Traditional Chinese Medicine, Southern Medical University were selected in this research, respectively, and all of these tissues were performed by immunohistochemistry (IHC) with a polyclonal antibody for LGR4. Meanwhile, from Aug, 2013 to Mar, 2019, 15 cases of serous ovarian cancer fresh tissues and 15 cases of paratumor fresh tissues who were operated at Integrated Hospital of Traditional Chinese Medicine, Southern Medical University were performed with Quantitative Real-time PCR to detect the mRNA expression of LGR4, respectively. RESULTS: LGR4 expression was much higher both in paraffin-embedded and fresh cancer tissues than that in paratumor tissues, respectively, and its expression was associated with recurrence free survival and overall survival in serous ovarian cancer patients. Moreover, in a multivariate model LGR4 was an indeed independent predictor of poor survival in serous ovarian cancer patients. CONCLUSION: LGR4 is upregulated in serous ovarian cancer, and LGR4 is an indeed useful independent prognostic predictor in serous ovarian cancer, and it may provide important clinical value of serous ovarian cancer.

Preparation of Borax Cross-Linked Starch Nanoparticles for Improvement of Mechanical Properties of Maize Starch Films.

Recently, starch nanoparticles have attracted widespread attention from various fields. In this study, a new strategy for preparing covalent-cross-linked starch nanoparticles was developed using boron ester bonds formed between debranched starch (DBS) and borax. The nanoparticles were characterized by Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), X-ray diffraction (XRD), dynamic light scattering (DLS), differential scanning calorimeter (DSC), and thermogravimetric analysis (TGA). The obtained nanoparticles were spherical with a size of 100-200 nm. The formation of boron ester bonds was confirmed by FTIR. The as-prepared starch nanoparticle exhibited a low relative crystallinity of 13.6%-23.5%. Compared with pure starch film, the tensile strength of starch film with 10% starch nanoparticles increased about 45%, and the elongation at break percentage of starch film with 5% starch nanoparticles increased about 20%. The new strategy of forming starch nanoparticles by using boron ester bonds will advance the research of carbohydrate nanoparticles.

High-throughput synapse-resolving two-photon fluorescence microendoscopy for deep-brain volumetric imaging in vivo.

Optical imaging has become a powerful tool for studying brains in vivo. The opacity of adult brains makes microendoscopy, with an optical probe such as a gradient index (GRIN) lens embedded into brain tissue to provide optical relay, the method of choice for imaging neurons and neural activity in deeply buried brain structures. Incorporating a Bessel focus scanning module into two-photon fluorescence microendoscopy, we extended the excitation focus axially and improved its lateral resolution. Scanning the Bessel focus in 2D, we imaged volumes of neurons at high-throughput while resolving fine structures such as synaptic terminals. We applied this approach to the volumetric anatomical imaging of dendritic spines and axonal boutons in the mouse hippocampus, and functional imaging of GABAergic neurons in the mouse lateral hypothalamus in vivo.

Fabrication and Characterization of Starch Nanohydrogels via Reverse Emulsification and Internal Gelation.

Biopolymer-based nanohydrogels have great potential for various applications, including in food, nutraceutical, and pharmaceutical industries. Herein, starch nanohydrogels were prepared for the first time via reverse emulsification coupled with internal gelation. The effects of starch type (normal corn, potato, and pea starches), amylose content, and gelation time on the structural, morphological, and physicochemical properties of starch nanohydrogels were investigated. The diameter of starch nanohydrogel particles was around 100 nm after 12 h of retrogradation time. The relative crystallinity and thermal properties of starch nanohydrogels increased gradually with an increasing amylose content and gelation time. The swelling behavior of starch nanohydrogels was dependent upon the amylose content, and the swelling ratios were between 2.0 and 14.0, with the pea starch nanogels exhibiting the lowest values and the potato starch nanogels exhibiting the highest values.

An update on pharmacotherapy of autism spectrum disorder in children and adolescents.

To date, no medication is proven to be effective in treating core symptoms of autism spectrum disorder (ASD). Psychotropic medications are widely used to target emotional and behavioural symptoms in ASD. This article reviewed evidence for pharmacotherapy, novel therapeutic agents, and Complementary and Alternative Medicine (CAM) in children and adolescents with ASD. Currently, only risperidone and aripiprazole have been approved by the US Food and Drug Administration (FDA) for treatment of irritability associated with ASD in children and adolescents. However, associated metabolic side-effects are concerning. Evidence supports use of methylphenidate and atomoxetine for attention deficit hyperactivity disorder (ADHD) symptoms and clonidine and guanfacine ER appear to be helpful. SSRIs are poorly tolerated and lack evidence in reducing restricted repetitive behaviours (RRB), anxiety, and depression. Buspirone shows promise in the treatment of RRB. The evidence is inconsistent for the effectiveness of anti-epileptic medications. Recent studies of glutamatergic, Gamma-aminobutyric acid (GABA)ergic, and cholinergic agents and oxytocin show inconsistent results. Despite wide use of CAM agents, the evidence is inconclusive. Melatonin can be helpful in reducing sleep problems. Overall, the evidence is limited for pharmacotherapy in children with ASD, and side-effects with long-term use can be burdensome.

Morphology and Characteristics of Starch Nanoparticles Self-Assembled via a Rapid Ultrasonication Method for Peppermint Oil Encapsulation.

Starch nanoparticles (SNPs) and peppermint oil (PO)-loaded SNPs were fabricated via an ultrasonic bottom-up approach using short linear glucan debranched from waxy maize starch. The effects of the glucan concentration, ultrasonic irradiation time, and chain length on the SNPs' characteristics were investigated. Under the optimal conditions, i.e., short linear glucan concentration of 5% and ultrasonication time of 8-10 min, SNPs were successfully prepared. The as-prepared SNPs showed good uniformity and an almost perfect spherical shape, with diameters of 150-200 nm. The PO-loaded SNPs also exhibited regular shapes, with sizes of approximately 200 nm. The loading capacity, encapsulation efficiency, and yield of PO-loaded SNPs were ∼25.5%, ∼87.7%, and ∼93.2%, respectively. After encapsulation, PO possessed enhanced stability against thermal treatment (80 °C). The pseudo-first-order kinetics model accurately described the slow-release properties of PO from SNPs. This new approach of fabricating SNPs is rapid, high yield, and nontoxic, showing great potential in the encapsulation and sustained release of labile essential oils or other lipids.

Effects of chitin nano-whiskers on the gelatinization and retrogradation of maize and potato starches.

Starch is very prone to retrogradation after gelatinization. Inhibition of starch retrogradation has been an important factor in improving the quality of food. For the first time, we investigated the effect of nano-materials, represented by chitin nano-whiskers (CNWs), on the short- and long-term retrogradation of maize and potato starches. Rapid Visco-Analyser results showed that the addition of CNWs significantly decreased the setback values of maize and potato starches, which suggested that CNWs could retard the short-term retrogradation of starch. Differential scanning calorimetry and X-ray diffraction results showed that the percentage of retrogradation of maize and potato starches significantly decreased (P<0.05), suggesting the inhibition of long-term retrogradation. The CNWs could be used as a new inhibitor of starch retrogradation to develop starch-based food with longer shelf life.

Pharmacotherapy for mental health problems in people with intellectual disability.

PURPOSE OF REVIEW: Psychotropic medications are commonly prescribed to people with intellectual disability. We reviewed current evidence-based pharmacotherapy options and recent updates to guide clinicians in their medication management plans. RECENT FINDINGS: Antipsychotics, particularly risperidone, appear to be effective in reducing problem behaviors in children with intellectual disability. Evidence in adults is inconclusive. Methylphenidate appears to be effective, and α-agonists appear promising in reducing attention-deficit hyperactivity disorder symptoms. Lithium might be effective in reducing aggression. Evidence is limited to support the use of antiepileptic drugs, anxiolytics, and naltrexone for management of problem behaviors. Antidepressants may be poorly tolerated and might not be effective in reducing repetitive/stereotypic behaviors.In recent trials, glutamatergic and GABAergic agents for fragile X syndrome, and acetylcholinesterase inhibitors for Down's syndrome, failed to show efficacy. Growth hormone treatment might improve cognition and behavior in Prader-Willi syndrome population. Results from oxytocin trials on social behaviors are inconclusive albeit promising. Melatonin appears to improve sleep. Most trials of dietary supplements did not show benefits. SUMMARY: Evidence-based pharmacotherapy options in people with intellectual disability are limited, and many agents can cause substantial adverse events. For this reason, clinicians should consider pharmacotherapy as only a part of comprehensive treatment, and regularly assess drug effects, adverse events, and the feasibility of decreasing dose or withdrawing medications.

Physiological concentrations of trans-11 18:1 vaccenic acid suppress pro-inflammatory markers under acute inflammation in isolated ICR mice splenocytes.

Direct influences of dietary trans-11 18:1 vaccenic acid (TVA) at physiological concentrations of 50-200 µM were evaluated for cell growth, cytotoxic activity, and cytokine production in leukocytes isolated from the mouse spleen. TVA supplementation for 24 h induced growth of splenocytes at concentrations of 50-200 µM, except for 100 µM. The cytokines TNFα, IFNγ, and IL-10 of splenocytes were stimulated by 100 µM TVA. Induced production of TNFα in splenocytes challenged with lipopolisaccharides was suppressed by 100 µM TVA. Physiological levels of TVA had direct effects on growth and cytokine production in splenocytes. Further in vivo studies are needed to improve understanding of the precise influence of trans fatty acids on production of pro-inflammatory markers under acute inflammation conditions.

Thalamus provides layer 4 of primary visual cortex with orientation- and direction-tuned inputs.

Understanding the functions of a brain region requires knowing the neural representations of its myriad inputs, local neurons and outputs. Primary visual cortex (V1) has long been thought to compute visual orientation from untuned thalamic inputs, but very few thalamic inputs have been measured in any mammal. We determined the response properties of ∼ 28,000 thalamic boutons and ∼ 4,000 cortical neurons in layers 1-5 of awake mouse V1. Using adaptive optics that allows accurate measurement of bouton activity deep in cortex, we found that around half of the boutons in the main thalamorecipient L4 carried orientation-tuned information and that their orientation and direction biases were also dominant in the L4 neuron population, suggesting that these neurons may inherit their selectivity from tuned thalamic inputs. Cortical neurons in all layers exhibited sharper tuning than thalamic boutons and a greater diversity of preferred orientations. Our results provide data-rich constraints for refining mechanistic models of cortical computation.