Mitigation of monocyte inflammation by inhibition of sodium phosphate co-transporter with phosphonoformic acid and parthenolide in diabetic nephropathy uremia

Purpose: To investigate the effect of sodium phosphate co-transporter (Pit-1) on the regulation of monocyte inflammation in diabetic nephropathy uremia (DNU) patients and the underlying principles of inflammatory immune response during DNU pathogenesis.Methods: The levels of CD14+ CD16+ and Pit-1 on peripheral blood mononuclear cells (PBMC) were measured by flow cytometry. Serum C-reactive protein (CRP) and 25(OH)D3 were detected by immunoturbidimetry while IL-6 and MCP-1 were assayed with enzyme-linked immunoassay (ELISA). The amounts of vitamin D receptors (VDRs) and Pit-1 mRNA in human acute monocytic leukemia cell lines (THP-1) were determined by quantitative real-time polymerase chain reaction (qRT-PCR), while western blot was utilized for measurement of NF-κB p65 and p-STAT5.Results: Compared to the healthy group, DNU patients showed markedly higher CD14+CD16+, Pit-1, CRP, IL-6 and MCP-1, while 25(OH) D3 was reduced. Following stimulation with PFA or PTN, comparison with DNU group revealed that THP-1 monocytes showed a significant down-regulation of Pit-1 (1.34 ± 0.06 for PFA; 1.60 ± 0.25 for PTN; p < 0.05); NF-κB p65 (2.65 ± 0.25 for PFA; 3.88 ± 0.13 for PTN; p < 0.01), p-STAT5 (2.49 ± 0.10 for PFA; 3.03 ± 0.09 for PTN; p < 0.01) and a significant decrease in levels of IL-6 (55.38 ± 3.22 for PFA, 68.68 ± 6.01 for PTN; p < 0.05); MCP-1( 39.67 ± 3.62 for PFA; 52.62 ± 5.00 for PTN; p < 0.01), except for VDR (0.64 ± 0.15 for PFA, 0.43 ± 0.03 for PTN; p < 0.05) .Conclusion: The level of expression of Pit-1 has a positive correlation with the level of inflammatory monocytes, which indicates that Pit-1 can be used as a new biomarker for DNU diagnosis. In addition, since Pit-1 is connected to NF-κB and STAT5 signaling pathways which are critical to inflammatory immune response, development of drugs that target Pit-1 could be an approach in developing new strategies for DNU therapy.Keywords: Pit-1, Diabetic nephropathy, Uremia, Monocytes, Inflammation, NF-κB, STAT

Lifshitz effects on holographic pp-wave superfluid

In the probe limit, we numerically build a holographic $p$-wave superfluid model in the four-dimensional Lifshitz black hole coupled to a Maxwell-complex vector field. We observe the rich phase structure and find that the Lifshitz dynamical exponent $z$ contributes evidently to the effective mass of the matter field and dimension of the gravitational background. Concretely, we obtain the Cave of Winds appeared only in the five-dimensional anti-de Sitter~(AdS) spacetime, and the increasing $z$ hinders not only the condensate but also the appearance of the first-order phase transition. Furthermore, our results agree with the Ginzburg-Landau results near the critical temperature. In addition, the previous AdS superfluid model is generalized to the Lifshitz spacetime.Comment: 14 pages,5 figures, and 1 table, accepted by Phys. Lett.

Paris polyphylla extract inhibits proliferation and promotes apoptosis in A549 lung cancer cells

Purpose: To investigate the effect of Paris polyphylla extract (PPE) on proliferation and apoptosis in A549 human lung cancer cells.Methods: Morphological changes were examined by microscopy in A549 cells after exposure to PPE. Trypan blue staining of living cells was used to aid the construction of the cell growth curve after treatment with different concentrations of PPE. The influence of PPE on cell proliferation, apoptosis and cell cycle were determined by MTT assay. Protein expressions of key apoptosis-related enzymes were determined by immuno-cytochemical method.Results: PPE inhibited the growth of A549 lung cancer cells at a concentration range of 12.5 – 200.0 μg/mL. Flow cytometry revealed that PPE promoted apoptosis in A549 cells. The proportion of cells in G0/G1-phase increased significantly (p < 0.01), while the proportion of cells in S- and G2/M-phases decreased correspondingly, indicating that the cells were in G0/G1-phase arrest. Cell cycle arrest and apoptosis-inducing effect gradually increased with increase in PPE concentration. With increasing concentration of PPE, there was significant increase in the expressions of caspase-8, caspase-3 and caspase-9, but significant decrease in Ki-67, p21ras protein (p < 0.01).Conclusion: PPE exerts pronounced inhibitory activity on the proliferation of A549 lung cancer cells. It also induces apoptosis in A549 cells, most probably by a mechanism related to Ki-67 and p21 ras protein expression, and arrest of cell cycle in G0/G1-phase.Keywords: Paris polyphylla, Antitumor activity, Lung cancer, A549 cells, p21 ras protein expression, Caspase, Cell cycle arrest, Apoptosi

p21-Activated Kinase 1 (Pak1) Phosphorylates BAD Directly at Serine 111 In Vitro and Indirectly through Raf-1 at Serine 112

Cell survival depends on the balance between protective and apoptotic signals. When the balance of signals tips towards apoptosis, cells undergo programmed cell death. This balance has profound implications in diseases including cancer. Oncogenes and tumor suppressors are mutated to promote cell survival during tumor development, and many chemotherapeutic drugs kill tumor cells by stimulating apoptosis. BAD is a pro-apoptotic member of the Bcl-2 family of proteins, which can be phosphorylated on numerous sites to modulate binding to Bcl-2 and 14-3-3 proteins and inhibit its pro-apoptotic activities. One of the critical phosphorylation sites is the serine 112 (S112), which can be phosphorylated by several kinases including Pak1.We mapped the Pak phosphorylation sites by making serine to alanine mutations in BAD and testing them as substrates in in vitro kinase assays. We found that the primary phosphorylation site is not S112 but serine 111 (S111), a site that is sometimes found phosphorylated in vivo. In transfection assays of HEK293T cells, we showed that Pak1 required Raf-1 to stimulate phosphorylation on S112. Mutating either S111 or S112 to alanine enhanced binding to Bcl-2, but the double mutant S111/112A bound better to Bcl-2. Moreover, BAD phosphorylation at S111 was observed in several other cell lines, and treating one of them with the Pak1 inhibitor 2,2'-Dihydroxy-1,1'-dinaphthyldisulfide (IPA-3) reduced phosphorylation primarily at S112 and to a smaller extent at S111, while Raf inhibitors only reduced phosphorylation at S112.Together, these findings demonstrate that Pak1 phosphorylates BAD directly at S111, but phosphorylated S112 through Raf-1. These two sites of BAD serve as redundant regulatory sites for Bcl-2 binding

Post-Training Dephosphorylation of eEF-2 Promotes Protein Synthesis for Memory Consolidation

Memory consolidation, which converts acquired information into long-term storage, is new protein synthesis-dependent. As protein synthesis is a dynamic process that is under the control of multiple translational mechanisms, however, it is still elusive how these mechanisms are recruited in response to learning for memory consolidation. Here we found that eukaryotic elongation factor-2 (eEF-2) was dramatically dephosphorylated within 0.5–2 hr in the hippocampus and amygdala of mice following training in a fear-conditioning test, whereas genome-wide microarrays did not reveal any significant change in the expression level of the mRNAs for translational machineries or their related molecules. Moreover, blockade of NMDA receptors with MK-801 immediately following the training significantly impeded both the post-training eEF-2 dephosphorylation and memory retention. Notably, with an elegant sophisticated transgenic strategy, we demonstrated that hippocampus-specific overexpression of eEF-2 kinase, a kinase that specifically phosphorylates and hence inactivates eEF-2, significantly inhibited protein synthesis in the hippocampus, and this effects was more robust during an “ongoing” protein synthesis process. As a result, late phase long-term potentiation (L-LTP) in the hippocampus and long-term hippocampus-dependent memory in the mice were significantly impaired, whereas short-term memory and long-term hippocampus-independent memory remained intact. These results reveal a novel translational underpinning for protein synthesis pertinent to memory consolidation in the mammalian brain.</p

Three new Dioszegia species (Bulleribasidiaceae, Tremellales) discovered in the phylloplane in China

The genus Dioszegia is comprised of anamorphic basidiomycetous yeasts and is classified in the family Bulleribasidiaceae of the order Tremellales. Currently, 24 species have been described and accepted as members of the genus, although its diversity and global distribution have not been thoroughly investigated. In this study, yeasts were isolated from plant leaves collected in the Guizhou and Henan Provinces of China and identified through a combination of morphological and molecular methods. Phylogenetic analyses of the combined ITS and LSU sequences coupled with morphological studies revealed three novel species, D. guizhouensis sp. nov., D. foliicola sp. nov., and D. aurantia sp. nov., proposed here. Additionally, our phylogenetic analyses suggest that the recently discovered species D. terrae is a synonym of D. maotaiensis. This study presents detailed descriptions and illustrations of three new Dioszegia species and highlights distinctions between them and their close relatives. The findings of this study contribute to our knowledge of the biodiversity of Dioszegia, offering a foundation for future research

Changes in Volatile Profiles and Activity of Hydroperoxide Lyase and Alcohol Dehydrogenase During the Development of Cabernet Sauvignon Grapes (Vitis vinifera L.)

In this study we focused on the development of Cabernet Sauvignon grapes and investigated changes in theactivity of alcohol dehydrogenase (ADH) and hydroperoxide lyase (HPL) in different tissues. We sampledgrape skin at four, six, seven, eight, nine, 10, 12, 14 and 16 weeks after anthesis; developing flowers whenblooming at 0%, 5%, 50%, and 90%; and leaves at two and four weeks before anthesis and at two, four,six, eight, nine, and 10 weeks after anthesis. We also examined the type and fluctuation of volatile contents.ADH activity increased with the development of flowers and grape skins, which led to the increasing oftypes and concentration of alcohols. Low levels of 9-HPL led to low concentrations of C9 compounds.According to this paper, C6 compounds became abundant with the development of grape berries, while theactivity of 13-HPL kept at a low level in the flowers and grape skins. There might have been a high level of13-HPL activity from the end of flowering until fruit setting that we did not detect. Furthermore, similarC6 and C5 compounds were detected across all tissues, including hexanal, (E)-2-hexenal, (Z)-3-hexenal,(Z)-2-penten-1-ol, (Z)-3-hexen-1-ol, 1-hexanol and 3-hexen-1-ol. Generally speaking, the concentrations ofC6 and C5 compounds could be used as the criterion of maturation of the three grape tissues