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Understanding the structure-activity connection (SAR) with the Wittig effect via genetically-encoded substrates.
Sequence comparisons showed 38-40% identity between the yeast and plant PCNA proteins, and > 91% identity among the plant PCNA proteins, which clustered together in one phylogenetic group. The expression of the six plant PCNA proteins in the yeast two-hybrid system confirmed interactions with the CLCuMuV and PeLCV Rep proteins. Our results demonstrate that the interaction of begomovirus Rep proteins with eukaryotic PCNA proteins is strongly conserved, despite significant evolutionary variation in the protein sequences of both of the interacting partners.This work reports the amy1 gene cloning from Massilia timonae CTI-57, and its successful expression in Escherichia coli Rosetta™ (DE3) from the pTRCHis2B plasmid. The recombinant AMY1 protein had 47 kDa, and its modeled structure showed a monomer composed of three domains. An N-terminal domain with the characteristic (β/α)8-barrel structure of α-amylases, which contained the catalytic amino acid residues. The second domain was small, and the C-terminal domain was similar to those found in the barley α-amylase. A phylogenetic analysis demonstrated a high sequence identity of the studied protein with bacterial and plant α-amylases from the GH13_6 subfamily. This is the first characterized bacterial α-amylase from this glucoside hydrolase subfamily. Besides starch, the enzyme was also active against maltodextrin, amylopectin, and blocked p-nitrophenyl α-d-maltoheptaoside, but could not use β-cyclodextrin or 4-nitrophenyl α-d-glucopyranoside. The K M for highly pure grade soluble starch from potato and V max values were 0.79 mg/mL and 0.04 mg/min, respectively. The calcium ion showed to be essential for the purified enzyme's activity, while EDTA, molybdenum, cobalt, and mercury were strong inhibitors. The enzyme was almost fully active in SDS presence. The enzyme's optimal pH and temperature were 6.0 and 60 °C, respectively, and its denaturation T m was 79 °C. A TLC analysis revealed that glucose and maltose are products of the enzyme's action on starch. In conclusion, this work described the M. timonae GH13_6 subfamily α-amylase, which showed to be thermostable and anionic detergent-resistant.LncRNA HOXA11-AS functions as regulator of tumorigenesis of several human cancers. Fluorescein-5-isothiocyanate The present study was intended to explore its regulatory control in human skin cancer with emphasis on understanding the underlying molecular mechanism. The results showed significant (P  less then  0.05) upregulation of lncRNA HOXA11-AS transcript levels in human skin cancer tissues and cell lines. The knockdown of HOXA11-AS significantly (P  less then  0.05) inhibited the proliferation and colony formation of A375 and HMCB skin cancer cells. Flow cytometry showed that HOXA11-AS knockdown triggered arrest of the A375 and HMCB cells at G2/M check point of cell cycle by inhibiting the expression of cyclin B1. Additionally, western blot analysis showed that HOXA11-AS knockdown resulted in the deactivation of PI3K/AKT/mTOR signaling pathway. The silencing of HOXA11-AS significantly (P  less then  0.05) inhibited the migration and invasion of the A375 and HMCB skin cancer cells. This was also accompanied by increase in E-cadherin and decrease in N-cadherin expression. Collectively, the results indicate that lncRNA HOXA11-AS regulates the proliferation, migration and invasion of human skin cancer and may exhibit therapeutic potential in the treatment of skin cancer.Bio-inoculants play an important role for sustainable agriculture. Application of nanocompounds in the agriculture sector provides strength and is reported to enhance crop production but the combined effect of nanocompounds and plant growth-promoting rhizobacteria on plants has not been studied much. Therefore, the present study was planned to observe the effect of two plant growth promotory Bacillus spp. along with nanozeolite on maize under field conditions using a randomized block design. Combined treatment of nanozeolite and bio-inoculants promoted plant height, root length, fresh and dry weight of shoot and root, chlorophyll, carotenoids, total sugar, protein and phenol contents in maize significantly over control. Enhanced level of catalase, peroxidase, superoxide dismutase, phenols, alcohols and acid-esters in treated plants over control showed their role in stress management. An increase of 29.80% in maize productivity over control was reported in the combined treatment of Bacillus sp. and nanozeolite. Our results indicate that the application of bio-inoculants with nanozeolite showed a positive response on the health and productivity of maize plants. Hence, these may be used to enhance the productivity of different crops.
The online version contains supplementary material available at 10.1007/s13205-020-02561-2.
The online version contains supplementary material available at 10.1007/s13205-020-02561-2.This study summarizes the response of a hot spring cyanobacterium Fischerella sp. strain HKAR-14, under simulated light conditions of ultraviolet radiation (UVR), photosynthetically active radiation (PAR), PAR + UV-A (PA) and PAR + UV-A + UV-B (PAB). Exposure to UVR caused a decline in growth and Chl a while total carotene content increased under PA and PAB. Maximum photochemical efficiency of photosystem II (F v /F m) and relative electron transport rate decreased significantly in PA and PAB exposure. Higher non-photochemical quenching and lower photochemical quenching values were observed in UVR-exposed samples as compared to the control. Levels of intracellular reactive oxygen species (ROS) increased significantly in PAB and PA. link2 Fluorescence microscopic images showed an increase in green fluorescence, indicating the generation of ROS in UVR. The antioxidant machinery including superoxide dismutase, catalase and peroxidase showed an increase of 1.76-fold and 2.5-fold superoxide dismutase, 2.4-fold and 3.7-fold catalase, 1.83-fold and 2.5-fold peroxidase activities under PA and PAB, respectively. High-performance liquid chromatography equipped with photodiode array detector, electrospray ionization mass spectrometry, Fourier-transform infrared spectroscopy and nuclear magnetic resonance spectroscopy analyses reveal the occurrence of a single mycosporine-like amino acid, shinorine (λ max 332.3 ± 2 nm, m/z 333.1), with a retention time of 1.157 min. The electrochemical characterization of shinorine was determined by cyclic voltammetry. The shinorine molecule possesses electrochemical activity and represents diffusion-controlled process in 0.1 M (pH 7.0) phosphate buffer. An antioxidant assay of shinorine showed its efficient activity as antioxidant which increased in a dose-dependent manner.Ophiocordyceps sinensis (Berk.) is a fungus closely related to medicinal mushroom, which belongs to the family Ophiocordycipitaceae. It is a well-known and rich herbal source of bioactive active constituents. The medicinal mushroom has garnered worldwide attention owing to its multifarious bioactivities. This mushroom grows on the larva of ghost moths (Hepialidae) and produces fruiting bodies, which serve as a vital natural source of medicine and supplementary diets. On account of the diverse pharmacological and bioactive constituents present in O. sinensis, it has been established as a potential antioxidant, anticancer, antibacterial, anti-proliferative, anti-inflammatory agent that has been successfully used for treating several health issues, including hypoxia-related problems encountered by mountaineers, pilgrims, tourists and soldiers occurring at high-altitude regions such as acute mountain sickness (AMS), high-altitude pulmonary edema (HAPE), high-altitude cerebral edema (HACE), frostbite, chilblains, hypothermia, etc. The most important pharmacologically active compounds present in the O. sinensis include nucleobases and its derivatives (adenosine, cordycepin, 3-deoxyadenosine, AMP, GMP, UMP, guanosine, uridine), polysaccharides (mannose, glucose, galactose, rhamnose, arabinose, xylose, galactose), proteins, peptides and steroids. This article focuses on the various research endeavors undertaken to scientifically establish the medicinal properties of O. sinensis, highlighting the various principally active compounds, their pharmacological action, drug designing and development and future perspective for various health benefits.Chemokine (CC-motif) ligand 2 (CCL2) is an inflammatory cytokine that regulates the infiltration and migration of monocytes. It is highly expressed by both tumor and stromal cells and has been associated with tumorigenesis. However, the effect of the exogenous administration of CCL2 on ovarian cancer remains largely unknown. In this report, we attempted to establish an expression system in Escherichia coli to produce recombinant hCCL2. The recombinant plasmid containing the hCCL2 cDNA was prepared using the prokaryotic-expression plasmid pGEX-5X-3 and transformed into E. coli BL21. GST-hCCL2 was successfully induced by 0.1 mmol/L IPTG at 20 °C for 6 h, and the recombinant protein was purified using affinity chromatography. link3 The purified protein was identified by SDS-PAGE and Western Blot. In vitro experiments revealed that rhCCL2 promoted the proliferation of ovarian cancer cells and increased the levels of phosphorylation of MEK and ERK1/2, and the levels of JUN, RELB and NF-κB2 mRNA. Furthermore, inhibition of ERK signaling by treatment with PD98059 decreased ovarian cancer cell proliferation and levels of JUN, RELB, and NF-κB2 mRNA, indicating that exogenous rhCCL2 increased the proliferation of ovarian cancer cells, partially by activating the MAPK/ERK pathway, and by targeting JUN, RELB, and NF-κB2. Our study uncovered a promoting role of exogenous CCL2 on ovarian cancer cell proliferation through the MAPK/ERK signaling pathway, which may facilitate the discovery of more potential roles of CCL2 in ovarian cancer.
The online version contains supplementary material available at 10.1007/s13205-020-02571-0.
The online version contains supplementary material available at 10.1007/s13205-020-02571-0.Lentinus squarrosulus (Hed Khon Khao) is a source of bioactive polysaccharides. Three L. squarrosulus crude polysaccharides (LSPs) were subjected to cold water (LSP-CP), hot water (LSP-HP), and aqueous alkaline (LSP-AP) extractions, and their functional compositions and radical scavenging activities were compared. Synchrotron radiation FTIR (SR-FTIR) spectra and PCA plot analysis in the bio-regions (4000-400 cm-1) revealed that functional composition LSPs differ significantly (P  less then  0.05). All LSPs had lipids, protein, and polysaccharides such as β-glucan. The major monosaccharides in LSP-CP and LSP-HP are d-galactose, d-glucose, and d-mannose at different proportions, while LSP-AP contained mainly d-glucose. Also, fucose and xylose were present in all the LSPs. LSP-CP, LSP-HP and LSP-AP induced maximum 1,1-diphenyl-2- picrylhydrazyl (DPPH) radical scavenging activity of 78.93 ± 0.42% at 3 mg/mL, 79.16 ± 1.43% at 3 mg/mL and 65.26 ± 1.74% at 5 mg/mL, whiles on 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), their maximum activities were 98.94 ± 0.16% at 3 mg/mL, 97.42 ± 0.76% at 3 mg/mL and 47.24 ± 0.045% at 5 mg/mL, respectively. The results showed that LSP-CP and LSP-HP are good ABTS scavengers, whiles LSP-AP is poor ABTS scavenger. In overall, LSPs consist of essential functional compositions and could be used as natural antioxidants. This exploitation of fungal fruiting body extracts increased the potential use of L. squarrosulus in food and medicinal industries.
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