Formation of sweet potato starch nanoparticles by ultrasonic-assisted nanoprecipitation: Effect of cold plasma treatment.

Starch nanoparticles (SNPs) were produced from sweet potato starches by ultrasonic treatment combined with rapid nanoprecipitation. The starch concentration, ultrasonic time, and the ratio of starch solution to ethanol were optimized through dynamic light scattering (DLS) technique to obtain SNPs with a Z-average size of 64.51 ± 0.15 nm, poly dispersity index (PDI) of 0.23 ± 0.01. However, after freeze drying, the SNPs showed varying degrees of aggregation depending on the particle size of SNPs before freeze-drying. The smaller the particle size, the more serious the aggregation. Therefore, we tried to treat SNPs with dielectric barrier discharge cold plasma before freeze drying. Properties including morphological features, crystalline structure and apparent viscosity of various starches were measured by field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), and rheometer, respectively. The results showed that, after cold plasma (CP) treatment, the aggregation of SNPs during freeze drying was significantly inhibited. Compared to the native sweet potato starch, SNPs showed a higher relative crystallinity and a lower apparent viscosity. After CP treatment, the relative crystallinity of CP SNPs was further higher, and the apparent viscosity was lower. This work provides new ideas for the preparation of SNPs and could promote the development of sweet potato SNPs in the field of active ingredient delivery.

Solid-state-cultured mycelium of Antrodia camphorata exerts potential neuroprotective activities against 6-hydroxydopamine-induced toxicity in PC12 cells.

Antrodia camphorata (A. camphorata) is an edible fungus containing various bioactive compounds generally used for health benefits. This study aimed to explore the potential neuroprotective activities of solid-state-cultured mycelium of A. camphorata (SCMAC) against Parkinson's disease (PD), as well as the underlying mechanism using an in vitro 6-hydroxydopamine (6-OHDA)-induced PC12 cell model. The results showed that SCMAC extracts alleviated cell toxicity induced by 6-OHDA and the loss of dopaminergic neurons, which was confirmed by the increase of cell viabilities, inhibition of cell apoptosis, the upregulation of tyrosine hydroxylase (TH) and dopamine transporter (DAT) levels and the downregulation of α-Synuclein level. After purification, 11 compounds were identified by the NMR technique, including a quinone, four phenolic acid derivatives, three ubiquinone derivatives, two alkaloids, and a triterpenoid. The present study suggests that SCMAC could be an attractive candidate for the prevention or treatment of PD. PRACTICAL APPLICATIONS: Parkinson's disease seriously affects the lifetime and quality of the elder population for a long history. Long-term consumption of L-DOPA will result in side effects, such as developing abnormal involuntary movements called dyskinesia. This study showed that natural SCMAC extracts could be a potential therapeutic agent for the treatment of neurodegenerative disorder.

Discovery of an unprecedented benz[α]anthraquinone-type heterodimer from a rare actinomycete Amycolatopsis sp. HCa1.

The angucylines are a family of aromatic polyketides featuring a tetracyclic benz[a]anthraquinone skeleton. This class of polycyclic aromatic polyketides are exclusively associated with actinomycetes and can undergo many modifications such as oxidation, ring cleavage, glycosylation and dimerization. Here we report the discovery of a new ether-linked benz[a]anthraquinone heterodimer, named mycolatone (1), from a grasshopper-derived actinomycete, Amycolatopsis sp. HCa1. The structure of mycolatone (1) was determined by comprehensive two-dimensional NMR analysis, high-resolution electrospray ionization mass spectrometry and biogenetic consideration. This new heterodimeric molecule is structurally derived from the dimerization of two tetracyclic angucylines, 2-hydroxy-5-O-methyltetragomycin and PD116779, through an ether bond between C-8 and C-8'. This new structural feature enrich the structural diversity of angucylines. Additionally, the surface tension activity and cytotoxic activities of 1 against human cervical cancer cell line (Hela), human gastric adenocarcinoma cell line (SGC-7901) and human lung adenocarcinoma cell line (SPC-A-1) were evaluated.

Highly Correlated Recurrence Prognosis in Patients with Metastatic Colorectal Cancer by Synergistic Consideration of Circulating Tumor Cells/Microemboli and Tumor Markers CEA/CA19-9.

Circulation tumor cells (CTCs) play an important role in metastasis and highly correlate with cancer progression; thus, CTCs could be considered as a powerful diagnosis tool. Our previous studies showed that the number of CTCs could be utilized for recurrence prediction in colorectal cancer (CRC); however, the odds ratio was still lower than five. To improve prognosis in CRC patients, we analyzed CTC clusters/microemboli, CTC numbers, and carcinoembryonic antigen (CEA)/carbohydrate antigen 19-9 (CA19-9) levels using a self-assembled cell array (SACA) chip system for recurrence prediction. In CRC patients, the presence of CTC clusters/microemboli may have higher correlation in metastasis when compared to the high number of CTCs. Additionally, when both the number of CTCs and serum CEA levels are high, very high odds ratios of 24.4 and 17.1 are observed in patients at all stages and stage III of CRC, respectively. The high number of CTCs and CTC clusters/microemboli simultaneously suggests the high chance of relapse (odds ratio 8.4). Overall, the characteristic of CTC clusters/microemboli, CEA level, and CTC number have a clinical potential to enhance CRC prognosis.

Fluoropyrimidin-2,4-dihydroxy-5-isopropylbenzamides as antitumor agents against CRC and NSCLC cancer cells.

A major cause of failure of therapy in patients with non-small cell lung cancer (NSCLC) is development of acquired drug resistance leading to tumor recurrence and disease progression. In addition to the development of new generations of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), different molecular targets may provide opportunities to improve the therapeutic outcomes. In this study, we utilized the core structure 5-fluorouracil (5-FU) or tegafur, a 5-FU prodrug combined through different linkers with resorcinol to generate a series of fluoropyrimidin-2,4-dihydroxy-5-isopropylbenzamides which inhibit potent Heat Shock Protein 90 (HSP90). These compounds were found to show significant antiproliferative activity in colorectal cancer (CRC) HCT116 and NSCLC A549, H460, and H1975 (EGFR L858R/T790 M double mutation) cells. Compound 12c, developed by molecular docking analysis and enzymatic assays exhibits promising inhibitory activity of HSP90. This compound, 12c shows the most potent HSP90 inhibitory activity with an IC50 value of 27.8 ± 4.4 nM, superior to that of reference compounds AUY-922 (Luminespib) and BIIB021 whose IC50 values are 43.0 ± 0.9 nM and 56.8 ± 4.0 nM respectively. This strong HSP90 inhibitory activity of 12c leads to rapid degradation of client proteins EGFR and Akt in NSCLC cells. In addition, 12c induces significant accumulation of a sub-G1 phase population in parallel with apoptosis by showing activated caspase-3, -8 and -9 and PARP induction. These results provide a new strategy for development of novel HSP90 inhibitors for cancer treatment.

Electrosprayed chitosan/alginate/polyvinyl alcohol nanoparticles as boric acid carriers for 10Boron neutron capture therapy.

Aim: To improve the killing efficacy of head and neck squamous cells (SAS) by boric acid-mediated boron neutron capture therapy (BNCT). Materials & methods: Boric acid-containing chitosan/alginate/polyvinyl alcohol nanoparticles (B-capNPs) were manufactured using the nano-electrospray process. Results: Less than 10% of the boric acid leaked from the B-capNPs over 2 days. The B-capNPs killed up to 2.8-fold more SAS cells and reduced cytotoxicity tenfold when compared with pure boric acid alone. B-capNPs show selective uptake in tumor cells with tumor/normal ratios of SAS to normal (NIH 3T3) and macrophage (RAW 264.7) cells of 4.0 and 3.5, respectively, which are greater than the minimum acceptable tumor/normal ratio for BNCT of 2.5. Conclusion: These findings illustrate that B-capNPs may be more superior as BNCT drugs than pure boric acid.

Design, synthesis, and evaluation of N-phenyl-4-(2-phenylsulfonamido)-benzamides as microtubule-targeting agents in drug-resistant cancer cells, displaying HDAC inhibitory response.

Microtubule-targeting agents (MTA) have enjoyed significant clinical success for decades. However, several mechanisms may cause inactivation of such drugs, leading to acquired resistance in patients treated with them. Therefore, drugs containing a stilbene-like skeleton and possessing dual inhibitory activity may provide a new and differentiated treatment for patients to overcome challenging acquired resistance. A new compound (16c) displays promising anticancer activity with GI50 of 22 ± 2 and 12 ± 0.1 nM in vincristine-resistant nasopharyngeal (KB-Vin) cancer cells and etoposide-resistant nasopharyngeal (KB-7D) cancer cells and is better than vincristine, etoposide, ABT-751, and MS-275. A mechanistic study revealed that 16c interferes with the cell cycle distribution and induces cell cycle arrest at the G2/M phase and severe mitotic spindle defects followed by apoptosis. In addition, it produces much more significant cytotoxicity than vincristine and etoposide in the corresponding resistant cells, indicating that it may be a promising candidate to overcome drug resistance in cancer cells. Compound 16c also displays inhibitory activity against HDAC 1 and HDAC 2 with IC50 values of 1.07 µM, and 1.47 µM, respectively. These findings may lead to a new type of structural motif for future development of drugs that could overcome acquired resistance to MTAs.

Nitrogen-doped carbon nanodots prepared from polyethylenimine for fluorometric determination of salivary uric acid.

Stable and low-cost carbon dots (C-dots) were prepared from polyethylenimine (PEI) by a hydrothermal method. It is found that the fluorescence of the C-dots (best measured at excitation/emission wavelengths of 365/473 nm) is quenched by selective oxidation of surface PEI by periodate but recovers in the presence of uric acid (UA). It is assumed that this is due to the selective reduction of the nitrone groups to hydroxylamine groups by UA. The findings were used to design a fluorometric method for determination of UA that has a 2.3 nM detection limit. This is lower than that of reported fluorometric and enzymatic assays. The performance of the method has been validated by determination of UA in samples of human saliva. It is found that the results agree well with those obtained by a commercial UA assay. Graphical abstract Schematic presentation of the polyethylenimine (PEI) carbon nanodots (C-dots) as a fluorescent probe for uric acid. Their fluorescence is quenched by periodate (IO4-) due to oxidative formation of nitrone groups, an subsequently restored due to reduction by uric acid (UA).

1,4-Naphthoquinones as inhibitors of Itch, a HECT domain-E3 ligase, and tumor growth suppressors in multiple myeloma.

A series of 1,4-naphthoquinones (10a-10q) were synthesized and evaluated for anticancer activity. Compound 10e was identified as an inhibitor of Itch, a HECT domain-E3 ligase. In an evaluation of in vivo efficacy, 10e exhibited remarkable anticancer activity with TGI values of 98.3% and 100% at 25 mg/kg and 50 mg/kg orally daily, respectively, against human RPMI-8226 multiple myeloma xenograft. Treatment with 10e also showed a decrease of Itch level in human RPMI-8226 multiple myeloma cells. Thus 10e is a lead compound for further development of inhibitors targeting E3 ligase for treatment of multiple myeloma.

Aptamer-conjugated polymeric nanoparticles for the detection of cancer cells through "turn-on" retro-self-quenched fluorescence.

We have developed a simple, sensitive, and rapid fluorescence assay for the detection of cancer cells, based on "turn-on" retro-self-quenched fluorescence inside the cells. 1,3-Phenylenediamine resin (DAR) nanoparticles (NPs) containing rhodamine 6G (R6G) are conjugated with aptamer (apt) sgc8c to prepare sgc8c-R6GDAR NPs, while that containing rhodamine 101 (R101) are conjugated with TD05 for the preparation of TD05-R101DAR NPs. The sgc8c-R6GDAR and TD05-R101DAR NPs separately recognize CCRF-CEM and Ramos cells. The fluorescence intensities of the two apt-DAR NPs are both weak due to self-quenching, but they increase inside the cells as a result of release of the fluorophores from the apt-DAR NPs. The apt-DAR NPs' structure becomes less compact at low pH value, leading to the release of the fluorophores. The sgc8c-R6GDAR and TD05-R101DAR NPs allow detection of as low as 44 CCRF-CEM cells and 79 Ramos cells mL(-1), respectively, using a commercial reader within 10 min. Practicality of the two probes have been validated by the quantitation and identification of CCRF-CEM and Ramos cells spiked in blood samples through conventional fluorescence and flow cytometry analysis, with advantages of sensitivity, selectivity, and rapidity.

Functional microgels assisted tryptic digestion and quantification of cytochrome c through internal standard mass spectrometry.

Quantitation of cytochrome c (Cyt c) in cell lysates through surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using gold nanoparticles (Au NPs) as the matrix and GR-10 peptide as an internal standard has been demonstrated. To shorten digestion time, temperature sensitive microgels containing trypsin (TR) and Au NPs have been employed. As-prepared functional microgels (TR/Au NPs/MGs) allow digestion of Cyt c within 15 s under microwave irradiation. The internal standard SALDI-MS approach provides linearity (R(2) = 0.98) of MS signal ratio (I 1168.6/I 1067.6) of the tryptic digested peptide (m/z 1168.6) to GR-10 peptide (m/z 1067.6) against the concentration of Cyt c ranging from 25 to 200 nM, with a limit of detection (at a signal-to-noise ratio of 3) of 10 nM. This approach has been validated by the analysis of the lysates of HeLa cells, with an average concentration of 13.7 ± 3.5 µM for cytoplasmic Cyt c. Increased concentrations of Cyt c in the HeLa cells treated with etoposide (a commercial drug) or carbon dots (potential drug) have been revealed through this simple, sensitive, and rapid SALDI-MS approach, supporting the drugs induced Cyt c-mediated apoptosis of the cells. This study has shown that this internal standard SALDI-MS approach holds great potential for cell study.

Mechanistic aspects of lauryl gallate-induced differentiation and apoptosis in human acute myeloid leukemia cells.

Lauryl gallate (LG) is a gallic acid derivative that has been widely used as an antioxidant food additive. In this study, we examined the anticancer effects of LG on the human acute myeloid leukemia (AML) HL60 and KG-1 cells. Our results showed that LG inhibited cell proliferation in a concentration- and time-dependent manner in both HL60 and KG-1 cells. The IC50s of LG in HL60 and KG-1 cells were 3.5 and 8.0 µM, respectively. Treatment with LG increased the proportions of annexin V-stained and sub-G1-phase HL60 and KG-1 cells. Moreover, activation of both extrinsic and intrinsic apoptotic pathways was involved in LG-induced AML cell apoptosis, accompanied by dissipation of mitochondrial membrane potential, downregulation of anti-apoptotic proteins (Bcl-2, Mcl-1, and Bcl-xL), upregulation of pro-apoptotic proteins (Bak, PUMA, DR4, and DR5), and increased caspase-2, -3, -8, and -9 activation. Our results also indicated that LG could induce monocytic differentiation in both HL60 and KG-1 cells, confirmed by morphological changes, nitroblue tetrazolium reduction assays, nonspecific esterase assays, and increased CD14 expression. After blocking LG-induced ERK and Sp1 expression using the ERK-specific inhibitor PD98059, monocytic differentiation in both HL60 and KG-1 cells decreased, suggesting that LG-induced differentiation proceeded through an ERK/Sp1 signaling axis.

Sensitive and selective DNA probe based on "turn-on" photoluminescence of C-dots@RGO.

In this study, highly hydrophilic and photoluminescent sheets of reduced graphene oxide decorated with carbon dots (C-dots@RGO), methylene blue (MB), and a probe DNA have been used for the detection of DNA. The photoluminescence of C-dots@RGO is quenched by MB, which is restored in the presence of a target DNA. The combination of the C-dots@RGO, MB, and a DNA probe is selective for perfectly matched DNA over mismatched DNA, mainly because relative to single-stranded DNA, double-stranded DNA intercalates more strongly with MB, but interacts more weakly with RGO. In the presence of a target DNA, MB intercalates with the as-formed double-stranded DNA and is released from the surface of C-dots@RGO, leading to "turn-on" photoluminescence. The practicality of this assay has been validated by the determination of tumor suppressor gene BRCA1, with linearity over the concentration range from 25 to 250 nM and a limit of detection (LOD, at a signal-to-noise ratio of 3) of 14.6 nM. The C-dots@RGO probe provides higher specificity towards target DNA than towards common salts, carbohydrates, amino acids, and proteins found in real samples. Having the advantages of simplicity, cost-effectiveness, selectivity, and sensitivity, the DNA-P/C-dots@RGO-MB probe on microwells has been successfully employed for the detection of DNA, suggesting its potential for multiple analyses of DNA targets when various DNA probes are employed.

Carbon dots prepared from ginger exhibiting efficient inhibition of human hepatocellular carcinoma cells.

Fluorescent carbon nanodots (C-dots; 4.3 ± 0.8 nm) from fresh tender ginger juice provide high suppression of the growth of human hepatocellular carcinoma cells (HepG2), with low toxicity to normal mammary epithelial cells (MCF-10A) and normal liver cells (FL83B). The inhibition is selective to HepG2 over other tested cancer cells, including human lung cancer cell line (A549), human breast cancer cell line (MDA-MB-231), and human cervical cancer cell line (HeLa). Western blot results reveal that the C-dots up-regulate the expression of p53 protein only in the HepG2 cell line. The 50% inhibiting concentration (IC50) value of the C-dots on HepG2 cells is 0.35 mg mL-1. Image cytometry results show significant uptake of C-dots by HepG2 cells that induce intracellular production of reactive oxygen species (ROS, 18.2-fold increased), while other cells remain almost the same in ROS levels after treatment with C-dots (1.11 mg mL-1). The C-dots trigger the pro-apoptotic factor to promote HepG2 cell apoptosis. The C-dots effectively inhibit the growth of tumors in nude mice (104 ± 14 vs. 3.7 ± 0.2 mg with and without treatment within 14 days).

[Identification and diagnosis of myocardial damage and acute myocardial infarction during cardiopulmonary resuscitation].

OBJECTIVE: To observe the change in contents of creatine kinase isoenzyme MB (CK-MB) and cardiac troponin I (cTnI) in peripheral blood, the elevation of ST in electrocardiogram, and the result of coronary arteriography, to identify myocardial damage and acute myocardial infarction during cardiopulmonary resuscitation (CPR). METHODS: Twenty-six patients with sudden cardiac arrest received CPR, and those patients who had blood circulation maintained for over 24 hours were included. The expression of CK-MB and cTnI activation in peripheral blood were determined at 0, 4, 8, 12, 16 and 20 hours after CPR in all patients. Electrocardiogram was checked every 2 hours in all patients. If CK-MB, cTnI and ST segment of electrocardiogram was higher than usual, or myocardial infarct with suspicious elevation of ST (STEMI), coronary arteriography and interventional therapy were carried out immediately. Patients were divided into three groups. The patients who were not found to have coronary artery block were classified as group A (15 cases), those who were found to have coronary artery block were group B (6 cases), and the remaining patients in whom ST segment of electrocardiogram did not elevate, and coronary arteriography and interventional therapy were not consider were classified as group C (5 cases). Control group consisted of 15 healthy people (group D). The change in CK-MB and cTnI in peripheral blood and the elevation of electrocardiogram ST segment were analyzed. RESULTS: In group A, CK-MB level began to elevate at CPR 4 hours, and it peaked at CPR 12 hours. cTnI began to raise at CPR 4 hours, peaking at CPR 16 hours, then decreased gradually. Elevation of ST was seen in more than two leads in electrocardiogram at the beginning of restoration of spontaneous circulation (ROSC), then lowered quickly, and the decrease exceeded 50% of the elevation at ROSC 2 hours. In group B, the levels of CK-MB and cTnI began to increase at CPR 4 hours, and remained elevated at CPR 20 hours. ST segment was elevated in more than two leads in electrocardiogram at the beginning of ROSC, and remained elevated after ROSC 2 hours. In group C, the CK-MB and cTnI concentrations were increased 4 hours after successful CPR, and reached peak at CPR 12, 16 hours respectively, then they decreased. ST segment of electrocardiogram was not elevated. In group D, the CK-MB and cTnI concentration was in the normal range. ST segment of electrocardiogram was not elevated. CONCLUSION: All patients manifested myocardial damage after CPR. Some patients showed STEMI after CPR. CK-MB and cTnI concentrations increased gradually after successful CPR without specificity for earlier identification of myocardial damage and STEMI. It is necessary to find a new reliable marker to check for myocardial damage. Relatively speaking, elevation of the ST segment in electrocardiogram has more predictive value. A decrease exceeds 50% of the elevation of ST segment in electrocardiogram at ROSC 2 hours, or the peak of contents of CK-MB and cTnI appear at CPR 12 hours or 16 hours indicates myocardial damage. If the elevation of ST segment does not descend after ROSC 2 hours, or the levels of CK-MB and cTnI remain elevated at CPR 20 hours, STEMI should be suspected, and it is necessary to undertake interventional therapy or thrombolysis therapy.

Improvement of recombinant baculovirus infection efficiency to express manganese superoxide dismutase in silkworm larvae through dual promoters of Pph and Pp10.

The silkworm, Bombyx mori, has been used as an important bioreactor for the production of recombinant proteins through baculovirus expression system (BES). There are several problems which will probably be the bottleneck for practical and industrial utilization of silkworm bioreactor. Traditionally, the recombinant virus should infect the larvae through individual dorsal injection by a syringe. This is a time- and labor-consuming procedure. This drawback has become a bottleneck for practical and industrial utilization of baculovirus expression system in the silkworm bioreactor. In this paper, we constructed a dual expression baculovirus to express the renovated polyhedron and target manganese superoxide dismutase (SOD) gene under P10 and polyhedron promoters, respectively, through oral infection. The results showed that the direct injection of recombinant rBacmid/BmNPV/SOD DNA with cellfectin reagent infected the silkworm larvae partially. When next batches of larvae were fed orally with hemolymph, which was collected from first batch of injected and infected larvae, the obvious symptom of infection was found and high target SOD was expressed. These results imply it is feasible to express target genes through combination of recombinant bacmid DNA injection and oral feeding by a dual expression bacmid baculovirus.

Human ribosomal protein L7 displays an ER binding property and is involved in ribosome-ER association.

Human ribosomal protein L7 incorporates an ER-binding characteristic. It is evident from the in vivo ER co-localization of the transiently expressed recombinant L7 in mycophenolic acid treated HeLa cells, the in situ detection of the fluorescent L7 at the ER in digitonin-permeablized HeLa cells, and the expression of a similar K(D) value to ribosomes binding to the ER. However, no ER co-localization and a lower K(D) was observed if the last 50 amino acid residues at the carboxyl end of L7 were removed, implying that the carboxyl region embodies the ER-binding specificity. Based on the inhibitory effect of an anti L7 antibody during ribosome rebinding to the microsome, we suggest that the L7-ER-binding nature could be one of multiple factors that allow a nascent peptide-less ribosome to remain at the ER.