Comprehensive Review of Ocular Angiostrongyliasis with Special Reference to Optic Neuritis

Angiostrongyliasis, caused by Angiostrongylus cantonensis infection, is a food-borne parasitic disease. Its larvae evoke eosinophilic inflammation in the central nervous system, but can also cause pathological changes in the eyes. Among ocular angiostrongyliasis cases, the incidence of optic neuritis is low and only few sporadic reports exist. Some patients with optic neuritis developed obvious hypopsia or even vision loss, which would seriously influence the quality of life of patients. Prompt treatment of optic neuritis caused by A. cantonensis is the key factor for minimizing the incidence of serious complications of this disease. In this review, we first provide a comprehensive overview of ocular angiostrongyliasis, and then focus on the clinical features of optic neuritis caused by A. cantonensis.     

Identification and characterization of P GCW14 : a novel, strong constitutive promoter of Pichia pastoris

The available promoters in the Pichia pastoris expression platform are still limited. We selected and identified a novel strong constitutive promoter, P _GCW14 , and tested its promoter activity using enhanced green fluorescent protein (EGFP) as a reporter. Potential promoter regions of P _GCW14 were cloned upstream of the EGFP gene and promoter activity was analyzed by measuring fluorescence intensity. P _GCW14 exhibited significantly stronger promoter activity than the classic strong constitutive promoters P _TEF1 and P _GAP under various carbon sources, suggesting that P _GCW14 is a strong and constitutive promoter. Hence, P _GCW14 can be used as a promoter for high-level expression of heterologous proteins.     

Rubia austrozhejiangensis sp. nov. (Rubiaceae) from Zhejiang,China

Rubia austrozhejiangensis Z. P. Lei, Y. Y. Zhou & R. W. Wang, a new species of Rubiaceae from China, is described and illustrated. The new species is similar to R. ovatifolia Z. Ying Zhang and R. argyi (H. Lév. & Vaniot) H. Hara ex Lauener, but differs from the former in having stems and branches cylindrical, not quadrate‐angled, long‐ovate to ovate‐lanceolate leaf blades, many‐flowered inflorescence, and smaller mericarps, 3–4 mm in diameter. In R. ovatifolia, stems and branches are quadrate‐angled, leaf blades ovate, ovate‐cordate to rounded cordate, and the inflorescences are sparsely flowered. Compared to R. argyi, the new species has cylindrical, not quadrate‐angled stems and branches, leaf blades that are long‐ovate to ovate‐lanceolate, 3–5‐veined, and slightly reflexed corolla lobes. In R. argyi, stems and branches are quadrate‐angled or winged, the corolla lobes are spreading, and the mericarps are 5–7 mm in diameter.     

9-Phenanthrol and flufenamic acid inhibit calcium oscillations in HL-1 mouse cardiomyocytes

It is well established that intracellular calcium ([Ca²⁺]_i) controls the inotropic state of the myocardium, and evidence mounts that a “Ca²⁺ clock” controls the chronotropic state of the heart. Recent findings describe a calcium-activated nonselective cation channel (NSC_Ca) in various cardiac preparations sharing hallmark characteristics of the transient receptor potential melastatin 4 (TRPM4). TRPM4 is functionally expressed throughout the heart and has been implicated as a NSC_Ca that mediates membrane depolarization. However, the functional significance of TRPM4 in regards to Ca²⁺ signaling and its effects on cellular excitability and pacemaker function remains inconclusive. Here, we show by Fura2 Ca-imaging that pharmacological inhibition of TRPM4 in HL-1 mouse cardiac myocytes by 9-phenanthrol (10 μM) and flufenamic acid (10 and 100 μM) decreases Ca²⁺ oscillations followed by an overall increase in [Ca²⁺]_i. The latter occurs also in HL-1 cells in Ca²⁺-free solution and after depletion of sarcoplasmic reticulum Ca²⁺ with thapsigargin (10 μM). These pharmacologic agents also depolarize HL-1 cell mitochondrial membrane potential. Furthermore, by on-cell voltage clamp we show that 9-phenanthrol reversibly inhibits membrane current; by fluorescence immunohistochemistry we demonstrate that HL-1 cells display punctate surface labeling with TRPM4 antibody; and by immunoblotting using this antibody we show these cells express a 130–150 kDa protein, as expected for TRPM4. We conclude that 9-phenanthrol inhibits TRPM4 ion channels in HL-1 cells, which in turn decreases Ca²⁺ oscillations followed by a compensatory increase in [Ca²⁺]_i from an intracellular store other than the sarcoplasmic reticulum. We speculate that the most likely source is the mitochondrion.     

The tumor suppressor CDKN3 controls mitosis

Mitosis is controlled by a network of kinases and phosphatases. We screened a library of small interfering RNAs against a genome-wide set of phosphatases to comprehensively evaluate the role of human phosphatases in mitosis. We found four candidate spindle checkpoint phosphatases, including the tumor suppressor CDKN3. We show that CDKN3 is essential for normal mitosis and G1/S transition. We demonstrate that subcellular localization of CDKN3 changes throughout the cell cycle. We show that CDKN3 dephosphorylates threonine-161 of CDC2 during mitotic exit and we visualize CDC2^pThr-161 at kinetochores and centrosomes in early mitosis. We performed a phosphokinome-wide mass spectrometry screen to find effectors of the CDKN3-CDC2 signaling axis. We found that one of the identified downstream phosphotargets, CKβ phosphorylated at serine 209, localizes to mitotic centrosomes and controls the spindle checkpoint. Finally, we show that CDKN3 protein is down-regulated in brain tumors. Our findings indicate that CDKN3 controls mitosis through the CDC2 signaling axis. These results have implications for targeted anticancer therapeutics.     

Involvement of CaMK‐IIδ and gelsolin in Cd2+‐dependent cytoskeletal effects in mesangial cells

Cadmium is a toxic metal with pleiotropic effects on cell death and survival. The mesangial cell is particularly responsive to Cd's effects on kinase signaling pathways and cytoskeletal dynamics. Here we show that CaMK‐II is a participant in the cytoskeletal effects of Cd²⁺. A major mesangial cell isoform, CaMK‐IIδ, was identified in pellets of DNase I pull‐downs and cytosolic immunoprecipitates of G‐actin. CaMK‐IIδ was also present in Triton X‐100‐insoluble cytoskeletal preparations and translocated to the cytoskeleton in a concentration‐dependent manner in Cd‐treated cells. Translocation was suppressed by KN93, an inhibitor of CaMK‐II phosphorylation. In vitro actin polymerization studies indicated that recombinant CaMK‐IIδ sequestered actin monomer. Cytoskeletal preparations from Cd‐treated cells decrease the rate of polymerization, but KN93 co‐treatment prevents this effect. Over‐expressed CaMK‐IIδ also translocated to the cytoskeleton upon Cd exposure, and this was prevented by KN93. Conversely, siRNA silencing of CaMK‐IIδ increases the effect of cytoskeletal extracts on actin polymerization, and abrogates the effect of Cd. The actin capping and severing protein, gelsolin, translocates to the cytoskeleton in the presence of Cd²⁺, dependent upon the phosphorylation of CaMK‐II, and is recovered together with actin and CaMK‐IIδ in G‐actin pull‐downs and F‐actin sedimentation. Translocation is accompanied by generation of a 50 kDa gelsolin fragment whose appearance is prevented by KN93 and CaMK‐IIδ silencing. We conclude that cytoskeletal effects of Cd in mesangial cells are partially mediated by Cd‐dependent activation of CaMK‐IIδ, binding of CaMK‐IIδ and gelsolin to actin filaments, and cleavage of gelsolin. J. Cell. Physiol. 228: 78–86, 2013. © 2012 Wiley Periodicals, Inc.     

In vivo effects of anacetrapib on preβ HDL: improvement in HDL remodeling without effects on cholesterol absorption

Cholesteryl ester transfer protein (CETP) transfers cholesteryl ester and triglyceride between HDL and apoB-containing lipoproteins. Anacetrapib (ANA), a reversible inhibitor of CETP, raises HDL cholesterol and lowers LDL cholesterol in dyslipidemic patients. We previously demonstrated that ANA increases macrophage-to-feces reverse cholesterol transport and fecal cholesterol excretion in hamsters, and increased preβ HDL-dependent cholesterol efflux via ABCA1 in vitro. However, the effects of ANA on in vivo preβ HDL have not been characterized. In vitro, ANA inhibited the formation of preβ, however in ANA-treated dyslipidemic hamsters, preβ HDL levels (measured by two-dimensional gel electrophoresis) were increased, in contrast to in vitro findings. Because changes in plasma preβ HDL have been proposed to potentially affect markers of cholesterol absorption with other CETP inhibitors, a dual stable isotope method was used to directly measure cholesterol absorption in hamsters. ANA treatment of hamsters (on either dyslipidemic or normal diet) had no effect on cholesterol absorption, while dalcetrapib-treated hamsters displayed an increase in cholesterol absorption. Taken together, these data support the notion that ANA promotes preβ HDL functionality in vivo, with no effects on cholesterol absorption.