Tyrosine phosphorylation of HPK1 by activated Src promotes ischemic brain injury in rat hippocampal CA1 region

AbstractHematopoietic progenitor kinase 1 (HPK1) is a hematopoietic cell-restricted member of the Ste20 serine/threonine kinase super family. We recently reported that HPK1 is involved in c-Jun NH2-terminal kinase (JNK) signaling pathway by sequential activation of MLK3–MKK7–JNK3 after cerebral ischemia. Here, we used 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo [3,4-d] pyrimidine (PP2) and MK801 to investigate the events upstream of HPK1 in ischemic brain injury. Immunoprecipitation and immunoblot results showed that PP2 and MK801 significantly decreased the activation of Src, HPK1, MLK3, JNK3 and c-Jun, respectively, during ischemia/reperfusion. Histology and TUNEL staining showed PP2 or MK801 protects against neuron death after brain ischemia. We speculate that this unique signaling pathway through the tyrosine phosphorylation of HPK1 promotes ischemic brain injury by activated Src via N-methyl-d-aspartate receptor and, ultimately, the activation of the MLK3–MKK7–JNK3 pathway after cerebral ischemia

(meso-5,7,7,12,14,14-Hexamethyl-1,4,8,11-tetra­azacyclo­tetra­deca-4,11-diene)nickel(II) bis­[O,O′-bis(4-methyl­phen­yl) dithio­phosphate]

In the title compound, [Ni(C16H32N4)](C14H14O2PS2)2 or [Ni(trans[14]dien)][S2P(OC6H4Me-4)2]2, where trans[14]dien is meso-5,7,7,12,14,14-hexa­methyl-1,4,8,11-tetra­azacyclo­tetra­deca-4,11-diene, the NiII ion lies across a centre of inversion and is four-coordinated in a relatively undistorted square-planar arrangement by the four N atoms of the macrocyclic ligand trans[14]dien. The two O,O′-di(4-methyl­phen­yl)dithio­phos­phates act as counter-ions to balance the charge. Important geometric data include Ni—N = 1.9135 (16) and 1.9364 (15) Å

Molecular and Functional Characterization of Odorant Binding Protein 7 From the Oriental Fruit Moth Grapholita molesta (Busck) (Lepidoptera: Tortricidae)

Odorant-binding proteins (OBPs) are widely and abundantly distributed in the insect sensillar lymph and are essential for insect olfactory processes. The OBPs can capture and transfer odor molecules across the sensillum lymph to odorant receptors and trigger the signal transduction pathway. In this study, a putative OBP gene, GmolOBP7, was cloned using specific-primers, based on the annotated unigene which forms the antennal transcriptome of Grapholita molesta. Real-time PCR (qRT-PCR) analysis revealed that GmolOBP7 was highly expressed in the wings of males and the antennae of both male and female adult moths, while low levels were expressed in other tissues. The recombinant GmolOBP7 (rGmolOBP7) was successfully expressed and purified via Ni-ion affinity chromatography. The results of binding assays revealed that rGmolOBP7 exhibited a high binding affinity to the minor sex pheromone 1-dodecanol containing Ki of 7.48 μM and had high binding capacities to the host-plant volatiles, such as pear ester, lauraldehyde and α-ocimene. RNA-interference experiments were performed to further assess the function of GmolOBP7. qRT-PCR showed that the levels of mRNA transcripts significantly declined in 1 and 2 day old male and female moths, treated with GmolOBP7 dsRNA, compared with non-injection controls. The EAG responses of dsRNA-injected males and females to pear ester, as well as the EAG responses of dsRNA-injected males to 1-dodecanol, were significantly reduced compared to the GFP-dsRNA-injected and non-injected controls. We therefore infer that GmolOBP7 has a dual function in the perception and recognition of the host-plant volatiles and sex pheromones

catena-Poly[[aqua­(dipyrido[3,2-a:2′,3′-c]phenazine-κ2 N 4,N 5)iron(II)]-μ-pyrazine-2,3-dicarboxyl­ato-κ3 N 1,O 2:O 3]

In the title compound, [Fe(C6H2N2O4)(C18H10N4)(H2O)]n, the FeII ion adopts a slightly distorted octahedral mer-FeN3O3 geometry, arising from one N,N′-bidentate dipyrido[3,2-a:2′,3′-c]phenazine ligand, one N,O-chelating pyrazine-2,3-dicarboxyl­ate dianion and one water mol­ecule. An O-bonded symmetry-related dianion completes the coordination of the metal. The bridging dianion results in a one-dimensional polymeric chain. Aromatic π–π stacking inter­actions between ligands [centroid–centroid separations = 3.528 (2) and 3.741 (2) Å] and O—H⋯O and O—H⋯N hydrogen bonds link the chains together, leading to a three-dimensional supra­molecular network

catena-Poly[[aqua­(pyrazino[2,3-f][1,10]phenanthroline-κ2 N 8,N 9)cobalt(II)]-μ-pyrazine-2,3-dicarboxyl­ato-κ3 N 1 O 2:O 3]

In the title compound, [Co(C6H2N2O4)(C14H8N4)(H2O)]n, the Co atom is bonded to one N,N′-bidentate pyrazino[2,3-f][1,10]phenanthroline (Pyphen) ligand, one N,O-bidentate pyrazine-2,3-dicarboxyl­ate (PZDC) dianion and one water mol­ecule in a distorted octa­hedral mer-CoN3O3 geometry. The CoII atoms are bridged by the PZDC dianions, forming an infinite one-dimensional chain running along the b axis. Adjacent chains pack together through π–π stacking inter­actions [centroid–centroid separations = 3.498 (4) and 3.528 (4) Å], and O—H⋯O and O—H⋯N hydrogen bonds involving the water mol­ecule complete the structure

Functional Analysis of the Chemosensory Protein GmolCSP8 From the Oriental Fruit Moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae)

Chemosensory proteins (CSPs) belong to a family of small water-soluble proteins that can selectively bind and transport odorant molecules for olfactory communication in insects. To date, their definite physiological functions in olfaction remain controversial when compared with odorant binding proteins (OBPs). To investigate the functions of CSPs in the oriental fruit moth Grapholita molesta, we determined the tissue expression patterns and binding properties of the CSP, GmolCSP8. The key binding sites of GmolCSP8 with a representative ligand were evaluated using molecular flexible docking, site-directed mutagenesis and ligand-binding experiments. Multiple sequence alignment and phylogenetic analysis showed that GmolCSP8 possesses a typical conserved four cysteines motif and shares high sequence identity with some CSP members of other Lepidopteran insects. GmolCSP8 was predominantly expressed in the wings and antennae of both male and female adults and may be involve in contact chemoreception. Recombinant GmolCSP8 (rGmolCSP8) exhibited specific-binding affinities to small aliphatic alcohols (C4–12) and had the strongest binding affinity to 1-hexanol. The three-dimensional structure of GmolCSP8 was constructed using the structure of sgCSP4 as a template. Site-directed mutagenesis and ligand-binding experiments confirmed that Thr27 is the key binding site in GmolCSP8 for 1-hexanol binding, because this residue can form hydrogen bond with the oxygen atom of the hydroxyl group in 1-hexanol, and Leu30 may play an important role in binding to 1-hexanol. We found that pH significantly affected the binding affinities of rGmolCSP8 to ligand, revealing that ligand-binding and -release by this protein is related to a pH-dependent conformational transition. Based on these results, we infer that GmolCSP8 may participate in the recognition and transportation of 1-hexanol and other small aliphatic alcohols

The Research of Biology Coupling Characteristics on the Shells of Haliotis discus hannai Ino

The surface morphologies, structures and materials of Haliotis discus hannai Ino shells were qualitatively studied by means of a stereoscopic microscope，a field emission scanning electronic microscopy, energy dispersive spectrometer and X-ray diffractometer, and abrasive particle wear was qualitatively and quantitatively studied by means of a pin-on-disc apparatus. The results showed that the outer layer surface of Haliotis discus hannai Ino shells was non-smooth and had some strumae or similar parallel convex wave. The shells of Haliotis discus hannai Ino are polycrystalline composites of calcium carbonate and proteins and glycoproteins and consist of the periostracum, prismatic and nacreous layers with calcite in the outer prismatic layer and aragonite in the inner nacreous layer. Nacreous layer is a natural composite comprised of calcium carbonate in the aragonite polymorph with organic macromolecules sandwiched in between, and the coupling of platelet interlocks and organic materials makes nacreous layer to be strong and tough.The abrasive particle wear tests showed that the abrasion resistance was different on the different parts of the shells, and the left of the shells possessed the highest abrasion resistance and the abrasion resistance of the shells was the lowest on the edge of the right. The nacreous layer possessed higher abrasion resistance than prismatic layer because of the coupling of structure and materials of nacreous layer. Key words: Haliotis discus hannai Ino shells; morphology; structure; materials; anti-wear; biological character; couplin

Hábitos alimenticios de Calanus sinicus (Crustacea: Copepoda) durante primavera y otoño en el mar Bohai, investigados mediante un índice de herbivoría

Pigment ingestion rate (PIR) and egg production rate (EPR) of the dominant copepod Calanus sinicus, as well as chlorophyll-a concentration and phytoplankton assemblages were measured in the Bohai Sea, North China in June 1997, October 1998 and May 1999. A herbivore index (H) was also calculated as the carbon specific ratio of PIR and EPR, in order to investigate its feeding habits in the spring and autumn phytoplankton bloom respectively. On average, chlorophyll-a concentration was relatively similar (1-1.34 mg m-3) in the three cruises, but PIR was quite different. It was 3.24 µg C female-1 d-1 in October, equivalent to one half of the PIR for June and one third of the PIR for May. Average EPR was highest in May, and quite similar during the other two months. According to H values, herbivorous feeding contributed 100% of the egg production of C. sinicus in June, 82.5% in May, but only 47.8% in October. It is possible that omnivorous feeding of C. sinicus in October was induced by a prevalence of large-sized diatoms and sufficient non-phytoplankton food resources during the autumn bloom period.La tasa de ingestión de pigmentos (PIR) y la tasa de producción de huevos (EPR) del copépodo dominante Calanus sinicus ha sido determinada, conjuntamente con la concentración de clorofila- a y la composición del fitoplancton, en el mar de Bohai, norte de China, en junio 1997, octubre 1998 y mayo 1999. Para investigar los hábitos alimenticios durante los blooms de fitoplancton de primavera y otoño se ha determinado un índice de herbivoría (H), calculado como el cociente de las tasas específicas PIR y EPR en carbono. En promedio, la concentración de clorofila-a fue relativamente similar (1-1,34 mg m-3) en las tres campañas oceanográficas. Pero los valores de PIR difirieron bastante, variando desde 3,24 μg C hembra-1 d-1 en octubre a valores un medio y un tercio de dicho valor, respectivamente, en junio y mayo. Los valores promedio de EPR fueron máximos en mayo, y bastante similares durante los otros dos meses. Según los valores del índice H obtenidos, la herbivoría contribuyó un 100% en la producción de huevos de C. sinicus en junio y un 82,5% en mayo, mientras que sólo explicó un 47,8% de la producción de huevos en octubre. este estudio sugiere que la alimentación omnivora de C. sinicus en octubre fue inducida por la prevalencia de diatomeas de gran tamaño y la presencia suficiente de alimento no fitoplanctónico durante el periodo del bloom otoñal.