Genetic and pharmacological inhibition of GRPR protects against acute kidney injury via attenuating renal inflammation and necroptosis.

Gastrin-releasing peptide (GRP) binds to its receptor (GRP receptor [GRPR]) to regulate multiple biological processes, but the function of GRP/GRPR axis in acute kidney injury (AKI) remains unknown. In the present study, GRPR is highly expressed by tubular epithelial cells (TECs) in patients or mice with AKI, while histone deacetylase 8 may lead to the transcriptional activation of GRPR. Functionally, we uncovered that GRPR was pathogenic in AKI, as genetic deletion of GRPR was able to protect mice from cisplatin- and ischemia-induced AKI. This was further confirmed by specifically deleting the GRPR gene from TECs in GRPRFlox/Flox//KspCre mice. Mechanistically, we uncovered that GRPR was able to interact with Toll-like receptor 4 to activate STAT1 that bound the promoter of MLKL and CCL2 to induce TEC necroptosis, necroinflammation, and macrophages recruitment. This was further confirmed by overexpressing STAT1 to restore renal injury in GRPRFlox/Flox/KspCre mice. Concurrently, STAT1 induced GRP synthesis to enforce the GRP/GRPR/STAT1 positive feedback loop. Importantly, targeting GRPR by lentivirus-packaged small hairpin RNA or by treatment with a novel GRPR antagonist RH-1402 was able to inhibit cisplatin-induced AKI. In conclusion, GRPR is pathogenic in AKI and mediates AKI via the STAT1-dependent mechanism. Thus, targeting GRPR may be a novel therapeutic strategy for AKI.

Discovery of a novel GRPR antagonist for protection against cisplatin-induced acute kidney injury.

The side effects of acute Kidney Injury (AKI) and nephrotoxicity limit the application of cisplatin in cancer treatment. Inflammation and oxidative stress paly important role in the pathogenesis of cisplatin-induced AKI. Gastrin-releasing peptide receptor (GRPR) plays an important role in inflammatory response. In this study, we designed 34 new Pd176252 analogs, most synthesized compounds could reduce cisplatin-induced HK2 cell death. Of these compounds, 9b had strong binding affinity with GRPR, and significantly increased HK2 cell viability. Compound 9b significantly downregulated the level of creatinine, blood urea nitrogen (BUN), and malondialdehyde (MDA), and recovered the glutathione (GSH) level in cisplatin-induced AKI model. It also decreased the level of kidney injury molecule-1(KIM-1) in vitro and vivo. In the further pathogenesis studies, 9b downregulated level of inflammatory factors (TNF-α, IL-1ß, IL-6 and MCP-1), suppressed the nuclear factor-kappa B (NF-kB) phosphorylation, and decreased GRPR level. The results suggested that ameliorating cisplatin-induced AKI actions of 9b was involved in downregulation of TNF-α, IL-1ß, IL-6, and MCP-1, inhibition of NF-kB activation, and reduction of GRPR and oxidative stress level.

Mutant breeding of Starmerella bombicola by atmospheric and room-temperature plasma (ARTP) for improved production of specific or total sophorolipids.

To enhance specific or total sophorolipids (SLs) production by Starmerella bombicola for specific application, mutant library consisting of 106 mutants from 7 batches was constructed via atmospheric and room-temperature plasma (ARTP). When compared to the wild strain, 11, 36 and 12 mutants performed increases over 30% in lactonic, acidic or total SLs production. Genetic stability investigation showed that 8, 7, and 4 mutants could maintain the improved SLs production capacity. Mutants of A6-9 and A2-8 were selected out for enhanced specific SLs and total SLs production in fed-batch cultivation in flask. Without optimization, A6-9 obtained the highest reported lactonic SLs production of 51.95 g/l and A2-8 performed comparable acidic and total SLs production of 68.75 g/l and 100.33 g/l with all the reported stains. The structural composition of the obtained SLs was analyzed by HPLC and LC/MS, and the results confirmed the enhancement of SLs and certain SL components. These mutants would be important in industrial applications because the production and purification costs of SLs could be greatly reduced. Besides, the acquisition of these mutants also provided materials for the investigation of regulation mechanism of SLs biosynthesis for further genetic engineering of S. bombicola. Furthermore, critical micelle concentration (CMC), minimum surface tension (STmin) and hydrophilic-lipophilic balance (HLB) of the SLs obtained from the wild and mutant strains were also examined and compared. These results demonstrated the feasibility of obtaining SLs with different properties from different strains and the high efficiency of mutation breeding of S. bombicola by ARTP.

Degradation and Transformation of Lignin by a Fungus Aspergillus Flavus Strain F-1.

BACKGROUND: Lignin is the largest natural aromatic polymer in nature and is also a unique aromatic-based biopolymer, accounting for nearly 30% of the earth's organic carbon. Generally, lignin is regarded as waste and is mainly used as a low- value fuel that is burned to generate heat and energy to solve the problem of biomass waste; for this obstacle of lignin, highly efficient biodegradation plays a critical role in developing an environmentally friendly technique for lignin biotransformation. OBJECTIVES: This study intends to isolate and purify several microbial strains from nature. It also explores how their lignin degradation is able to enhance the biodegradation and recycling of biomass and the reclamation of lignin in wastewater from pulp and paper mills. MATERIALS AND METHODS: Lignin-degrading microbial strains were isolated from soil using medium containing sodium lignosulphonate as the sole carbon source. They were then screened by aniline blue and guaiacol plate, and then the best strain was chosen and identified. The conventional one-factor method was used to optimize various parameters that affect lignin's degradation ability. RESULTS: The strain possessing the highest lignin biodegradation ability was identified and denominated as Aspergillus Flavus F-1. After optimization, the maximum degradation rate of lignin, 44.6% within 3 days, was obtained at pH 7.0, 30 ℃, 2.5 g·L-1 ammonium sulfate, 2 g·L-1 lignin and 0.5 g·L-1 glucose. The results show the LiP and Lac secreted from Aspergillus Flavus F-1 played the main role in the degradation of lignin. CONCLUSION: One microbial strain, Aspergillus Flavus F-1, was successfully isolated with a lignin-degrading ability that can cut the lignin into fragments. This provides a promising candidate for the transformation and utilization of crop waste biomass for various industrial purposes.

Dual Anti-cancer and Anti-Itch Activity of PD176252 Analogues: Design, Synthesis and Biological Evaluation.

BACKGROUND: Cancer patients treated with targeted anti-cancer drug suffer from itch or pruritus. Itch or pruritus is an unpleasant sensation that brings about a negative impact on quality of life, and serious itch may lead to dose reduction and even discontinuation. Gastrin releasing peptide receptor (GRPR) plays a critical role in itch, inflammation and cancer, and GRPR antagonist has obvious effect on cancer, inflammation and itch. The aim of this paper is to develop a new agent with anti-cancer and anti-itch activity. METHODS: A series of GRPR antagonist PD176252 analogues (3a-3l) were designed and synthesized. Both anticancer and anti-itch activities were evaluated. Anti-cancer activity was evaluated in three human cancer cell lines in vitro, the anti-itch activity in evaluated with Kunming mice by intrathecal injection of chloroquine phosphate as a modeling medium. And the cytotoxicity on normal cells was evaluated. RESULTS: Of the tested compounds, compound 3i showed potently anti-cancer activity to all cancer cell lines tested with IC50 values of 10.5µM (lung), 11.6µM (breast) and 12.8µM (liver) respectively and it also showed significant inhibition of the scratching behavior. Comparing with PD17625, compound 3i and 3g gave better inhibition activities against all cancer cell lines, compound 3b, 3c and 3i showed better anti-itch activity. The compound 3i is safe for normal breast and liver normal cells, but it has high cytotoxicity on normal lung cell. CONCLUSION: The synthesized compounds have dual anti-cancer and anti-itch activity, so the development of drug with dual anti-tumor and anti-itch property is possible.

Anti-inflammatory Effects of a Small Molecule Gastrin-Releasing Peptide Receptor Antagonist on Adjuvant-Induced Rheumatoid Arthritis in Rats.

The anti-inflammatory effects of (R)-2-(1H-Imidazol-1-yl) ethyl-3-(1H-indol-3-yl)-2-(2-p-tolylacetamido)propanamide (RH-1402), a previous designed small molecule Gastrin releasing peptide (GRP) antagonist were evaluated in adjuvant-induced arthritic model of rats, and the inhibitory effect on neutrophil migration induced by GRP was determined by a transwell system experiment in vitro. The arthritis was induced by injection of Complete Freund's Adjuvant (CFA) containing 10 mg/mL of heat killed mycobacterium into the left hind footpad. Experimental rats were randomly divided into 6 groups, including control, placebo, positive control group, RH-1402 of low/middle/high dose group. Disease incidence and severity was evaluated through scoring of the paw edema and histologic features of joint synovial. Blood of all experimental rats was collected for interleukin 1ß (IL-1ß) and tumor necrosis factor α (TNF-α) cytokine levels. A transwell system was used to investigate whether RH-1402 would inhibit neutrophils migrating up a gradient of GRP in vitro. RH-1402 (5 and 10 mg/kg) significantly decreased adjuvant induced increased arthritis index during the administration period (days 14-20). Significant inhibition of joint synovial histological features can be found in the RH-1402 treated group, including alleviated Hyperplasia, Inflammatory of infiltration and activation of pannus formation. It also suppressed TNF-α and IL-1ß level. Five and 10 mg/kg of RH-1402 significantly inhibited the effect of GRP on neutrophil migration with a dose dependent relationship. These findings indicate that RH-1402 have potential protective anti-inflammatory effects on experimental models of arthritis.

Fabrication of mesoporous lignin-based biosorbent from rice straw and its application for heavy-metal-ion removal.

Lignocellulosic biomass offers the most abundant renewable resource in replacing traditional fossil resources. However, it is still a major challenge to directly convert the lignin component into value-added materials. The availability of plentiful hydroxyl groups in lignin macromolecules and its unique three-dimensional structure make it an ideal precursor for mesoporous biosorbents. In this work, we reported an environmentally friendly and economically feasible method for the fabrication of mesoporous lignin-based biosorbent (MLBB) from lignocellulosic biomass through a SO3 micro-thermal-explosion process, as a byproduct of microcrystalline cellulose. BET analysis reveal the average pore-size distribution of 5.50nm, the average pore value of 0.35cm3/g, and the specific surface area of 186m2/g. The physicochemical properties of MLBB were studied by fourier transform infrared spectroscopy (FTIR), attenuated-total-reflection fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), and element analysis. These results showed that there are large amounts of sulfonic functional groups existing on the surface of this biosorbent. Pb(II) was used as a model heavy-metal-ion to demonstrate the technical feasibility for heavy-metal-ion removal. Considering that lignocellulosic biomass is a naturally abundant and renewable resource and SO3 micro-thermal-explosion is a proven technique, this biosorbent can be easily produced at large scale and become a sustainable and reliable resource for wastewater treatment.

Sophorolipids production from rice straw via SO3 micro-thermal explosion by Wickerhamiella domercqiae var. sophorolipid CGMCC 1576.

A novel lignocellulose material, holocellulose from rice straw via the pretreatment of SO3 micro-thermal explosion, was developed to produce sophorolipids (SLs) with Wickerhamiella domercqiae var. sophorolipid CGMCC 1576. The influence factors of inoculum dose, yeast extract concentration and pH regulators (chemical regents used for adjusting/influencing pH) was investigated and discussed. Results showed that W. domercqiae can grow in the rice straw holocellulose hydrolysate, and acquire relative high SL yield of 53.70 ± 2.61 g/L in shake flask culture. Inoculum dose, yeast extract concentration and pH regulator made obvious influence on fermentation parameters, especially on final broth pH and SLs production. Furthermore, there is a strong negative linear correlation existing between final broth pH and lactonic SL or ratio of lac SL/tot SL. Additionally, comparison between SL production and non-glucose carbon sources, culture methods, microbes in previous reports was carried out. These results will be benefit for acquiring SL mixture with suitable lac SL/tot SL ratio for specific purpose and scope economically.

Discovery and pharmacophore studies of novel pyrazole-based anti-melanoma agents.

Due to the rising incidence and lack of effective treatments, malignant melanoma is the most dangerous form of skin cancer, so that new treatment strategies are urgently needed. Several recent developments indicate that the V600E mutant BRAF (BRAF(V600E) ) is a validated target for antimelanoma-drug development. Based on in silico screening results, a series of novel pyrazole derivatives has been designed, synthesized, and evaluated in vitro for their inhibitory activities against BRAF(V600E) melanoma cells. Compound 3d exhibited the most potent inhibitory activity with an IC50 value of 0.63 µM for BRAF(V600E) and a GI50 value of 0.61 µM for mutant BRAF-dependent cells. Furthermore, the QSAR modeling and the docking simulation of inhibitor analogs provide important pharmacophore clues for further structural optimization.

Chemoselective synthesis of trifluoromethylated γ-butenolide derivatives via phosphine-promoted tandem reaction of allylic carbonates and trifluoromethyl ketones.

A novel chemoselective phosphine-mediated tandem reaction between nonsubstituted MBH carbonates and aryl trifluoromethyl ketones is described. The product selectivity of the reaction is easily tunable by changing the ratios of the two reactants, and mono- or bicyclic bistrifluoromethylated vinyl γ-butenolide products can be prepared with good chemoselectivity in modest-to-good yields and diastereoselectivities. The formation of the bicyclic γ-butenolide structures via a one-pot four-step sequence under phosphine catalysis is unprecedented.

Construction of supramolecular hyperbranched polymers via the "tweezering directed self-assembly" strategy.

A bis-alkynylplatinum(II) terpyridine tweezer-alkynylgold(III) diphenylpyridine guest is shown to maintain the specific complexation in the presence of a B21C7-secondary ammonium salt recognition motif, which facilitates the formation of supramolecular hyperbranched polymers via the "tweezering directed self-assembly" strategy.

Cooperative complexation of amino acid derivatives to platinum acetylide-based bolaamphiphile.

A platinum(II) acetylide-based bolaamphiphile equipped with two peripheral B18C6 moieties has been successfully prepared, which demonstrates cooperative recognition behavior toward l-alanine ester salt in chloroform. In a polar methanol/chloroform (3/1, v/v) medium, the amino acid additives influence the aggregation of the bolaamphiphile significantly, leading to the morphological transition from nanospherical to disordered structures. The adaptive properties of the current host-guest binary system will benefit the development of stimuli-responsive supramolecular materials.

HPMC/PAA hybrid nanogels via aqueous-phase synthesis for controlled delivery of insulin.

A facile controlled-release delivery system has been developed with hydroxypropylmethylcellulose/poly(acrylic acid) (HPMC/PAA) hybrid nanogels by surfactant-free polymerization in aqueous solution. The effect of temperature, reaction time, and amount of cross-linker on the size and morphology of the nanogels has been studied. The results showed that when the reaction temperature is higher than the lower critical solution temperature, the particle size increases with increasing temperature. And in a certain range, the particle size decreases with increasing reaction time and amount of cross-linker, the prepared nanogel under each condition being spherical. Thus, the HPMC/PAA hybrid nanogels were prepared at 41 °C for 4 h, and possessed a size ranging from 250 nm to 615 nm. Then, insulin was loaded into the nanogel, and the pH and temperature sensitivity of the nanogel was studied, the results indicating that the release behavior of the nanogel can be regulated by pH and temperature. Finally, in streptozotocin-induced diabetic mice, insulin-loaded HPMC/PAA complex was able to maintain blood glucose concentration at a low level for the first week compared to a control group. Therefore, the nanogels have potential biological application as a long-term, sustained insulin delivery system.

Phosphine catalyst-controlled cycloaddition or dienylation reactions of trifluoromethyl aryl ketones with bis-substituted allenoates.

A chemoselective phosphine-catalyzed cycloaddition or dienylation reaction between trifluoromethyl-substituted ketones and bis-substituted allenoates was described. Under the catalysis of triarylphosphine, the reaction gave a range of trifluoromethylated tetrahydrofurans with broad substrate tolerance and good to excellent stereoselectivity, while the use of trialkylphosphine switched the reaction pathway to furnish CF3-substituted dienyl tertiary alcohols chemoselectively. Moreover, a preliminary study on the asymmetric version of the reaction was also performed, which represents the first example of a phosphine-catalyzed asymmetric reaction between allenoates and carbonyl compounds.

A novel phospholipase D constitutively secreted by Ochrobactrum sp. ASAG-PL1 capable of enzymatic synthesis of phosphatidylserine.

Phospholipase D (PLD) plays a crucial role in the enzyme-mediated preparation of phosphatidylserine (PS). A new PLD constitutively secreted by Ochrobactrum sp. ASAG-PL1 is now reported for the first time. After purification, the MW of the enzyme was ~37 kDa by SDS-PAGE. Its activity was highest at 40 °C and pH 7.0 and more than 90 % of the initial activity was maintained after 30 days at 4 °C and pH 7.0. This enzyme may then be useful as a potential biocatalyst for PS production.

Synthesis and biological evaluation of some novel resveratrol amide derivatives as potential anti-tumor agents.

Three series of novel resveratrol amide derivatives (1a-q, 2a-h, 3a-l) were synthesized and evaluated for their biological activities. All compounds were characterized by (1)H NMR, (13)C NMR, MS and elemental analysis. Furthermore, compound 3e was also characterized by X-ray crystallography. All the compounds were evaluated for their anti-tumor activity against MCF-7, A549 and B16-F10 tumor cell lines as well as cyclooxygenase-2 (COX-2)-derived prostaglandin E2 (PGE2) inhibitory activity of murine macrophage RAW 264.7 cell line. Among them, compounds 1c, 1g and 3e displayed the most potent COX-2 inhibitory activity with the IC50 values of 1.02, 1.27 and 1.98 µM, respectively. Molecular docking studies were performed to position compounds 1c and 3e into the active site of COX-2 to determine the probable binding modes.

One-step fermentation of pretreated rice straw producing microbial oil by a novel strain of Mortierella elongata PFY.

A fungus strain producing microbial oils utilizing pretreated rice straw was isolated from soil. This strain was identified as Mortierella elongata PFY based on the morphology and internal transcribed spacer sequence. Using pretreated rice straw as substrate, the average yield of total lipids was 7.07% after 7 days fermentation. The GC-MS detection demonstrated that the lipids were composed of saturated fatty acids and polyunsaturated fatty acids. This work presents one new way to make the waste biomass (rice straw) valuable.

Design, synthesis and anti-itch activity evaluation of aromatic amino acid derivatives as gastrin-releasing peptide receptor antagonists.

Eight aromatic amino acid derivatives (9a-9h) as gastrin-releasing peptide receptor (GRPR) antagonists were designed and synthesized. For the design, the tertiary structure of GRPR was predicted by blast searching in the premise of 1u19 protein as the template. Eight target compounds were docked into the binding pocket to investigate their possible binding interactions. Their anti-itch activities were tested by intrathecal injection using Kunming mice as experimental animals and chloroquine phosphate as a modeling medium. Compounds 9e and 9f significantly inhibited scratching behaviors. The anti-itch activities of these compounds decreased with the following sequence: 9e > 9f = 9d > 9b > 9g > 9a > 9h > 9c predicted by computer-aided drug design (CADD) and 9e > 9f > 9b > 9h > 9g > 9d > 9a > 9c evaluated by preliminary test. They were broadly in line with activity order pretested by CADD. It showed that the predicted tertiary structure of GRPR could be used for antipruritic drug design.

Synthesis, biological evaluation and molecular docking studies of resveratrol derivatives possessing curcumin moiety as potent antitubulin agents.

A series of resveratrol derivatives possessing curcumin moiety were synthesized and evaluated for their antiproliferative activity against three cancer cell lines including murine melanoma B16-F10, human hepatoma HepG2 and human lung carcinoma A549. Among them, compound C5 displayed the most potent in vitro antiproliferative activity against B16-F10 with IC(50) value of 0.71 µg/mL. Compound C5 also exhibited good tubulin polymerization inhibitory activity with IC(50) value of 1.45 µg/mL. Furthermore, docking simulation was carried out to position C5 into the tubulin-colchicine binding site to determine the probable binding mode.

Structure and saccharification of rice straw pretreated with sulfur trioxide micro-thermal explosion collaborative dilutes alkali.

In this paper, a sulfur trioxide collaborative dilutes alkali method has been developed to pre-treat rice straw and it has been studied that the pre-treated rice straw structure affected the saccharification of the rice straw hydrolyzed by cellulose enzymatic hydrolysis. The results show that the reaction of the sulfur trioxide with rice straw resulted in the internal micro-thermal explosion, and the saccharification rate was 91% based on the pretreated rice straw with sulfur trioxide for 4h following 1% w/v NaOH treatment for 7h at 50°C.