Behavior and endocrine effects of 3,4,5-trimethoxyamphetamine in male mice

Summary Effects of single doses of 50 and 100 mg/kg of TMA given i.p. were noted in male albino mice after 40 min and 2 1/2 h. Locomotor activity was significantly altered and biochemical tests indicated stimulatory effects on adrenocortical and adrenomedullary functions due to TMA.     

The effect of thiazol-4-ylmethoxyamine,a histidine decarboxylase inhibitor,on the development of morphine tolerance and physical dependance in mice

Summary A new histidine decarboxylase inhibitor, thiazol-4-ylmethoxyamine (TMA), injected into mice in a dose of 100 mg/kg i.p. 48 h before the implantation of a morphine-containing pellet, inhibited the development of morphine tolerance and physical dependence.I thank the World Health Foundation (Hong Kong) for a grant in aid of this research.     

Recruitment of Pyk2 to SHPS-1 signaling complex is required for IGF-I-dependent mitogenic signaling in vascular smooth muscle cells

In vascular smooth muscle cells, IGF-I stimulates SHPS-1/SHP2/Src complex formation which is required for IGF-I-stimulated cell proliferation. Using SHP2/Src silencing and a Pyk2/Y402F mutant, we showed that Pyk2 was also recruited to the SHPS-1 complex. Pyk2 recruitment to SHPS-1 is mediated via the interaction of Pyk2 Tyr402 and the Src in response to IGF-I. Following Src/Pyk2 association, Src phosphorylates Pyk2 on Tyr881 providing a binding site for Grb2. Cells expressing Pyk2/Y881F showed decreased Grb2 recruitment to SHPS-1 and impaired Shc/Grb2 association. This change led to reduced Erk1/2 (MAP kinase) activation and cell proliferation in response to IGF-I. Our results show that, following its recruitment to the SHPS-1 signaling complex, Pyk2 localizes Grb2 in close proximity to Shc thereby facilitating Shc/Grb2 association which leads to Erk1/2 activation in response to IGF-I. Thus, Pyk2 recruitment to SHPS-1 plays an important role in regulating the IGF-I-stimulated mitogenic response.     

Neurogenic responses of resistance and capacitance vessels

- . , , , . , .     

Re-evaluation of protein kinase CK2 pleiotropy: new insights provided by a phosphoproteomics analysis of CK2 knockout cells

CK2 denotes a ubiquitous and pleiotropic protein kinase whose holoenzyme is composed of two catalytic (α and/or α′) and two regulatory β subunits. The CK2 consensus sequence, S/T-x-x-D/E/pS/pT is present in numerous phosphosites, but it is not clear how many of these are really generated by CK2. To gain information about this issue, advantage has been taken of C2C12 cells entirely deprived of both CK2 catalytic subunits by the CRISPR/Cas9 methodology. A comparative SILAC phosphoproteomics analysis reveals that, although about 30% of the quantified phosphosites do conform to the CK2 consensus, only one-third of these are substantially reduced in the CK2α/α′^(?/?) cells, consistent with their generation by CK2. A parallel study with C2C12 cells deprived of the regulatory β subunit discloses a role of this subunit in determining CK2 targeting. We also find that phosphosites notoriously generated by CK2 are not fully abrogated in CK2α/α′^(?/?) cells, while some phosphosites unrelated to CK2 are significantly altered. Collectively taken our data allow to conclude that the phosphoproteome generated by CK2 is not as ample and rigidly pre-determined as it was believed before. They also show that the lack of CK2 promotes phosphoproteomics perturbations attributable to kinases other than CK2.     

Resistance to β-lactam antibiotics

-lactams have a long history in the treatment of infectious diseases, though their use has been and continues to be confounded by the development of resistance in target organisms. -lactamases, particularly in Gram-negative pathogens, are a major determinant of this resistance, although alterations in the -lactam targets, the penicillin-binding proteins (PBPs), are also important, especially in Gram-positive pathogens. Mechanisms for the efflux and/or exclusion of these agents also contribute, though often in conjunction these other two. Approaches for overcoming these resistance mechanisms include the development of novel -lactamase-stable -lactams, -lactamase inhibitors to be employed with existing -lactams, -lactam compounds that bind strongly to low-affinity PBPs and agents that potentiate the activity of existing -lactams against low-affinity PBP-producing organisms.Received 9 February 2004; received after revision 30 March 2004; accepted 19 April 2004     

Polyribosome patterns in EMC-virus infected Krebs-2 cells and cell extracts

. -2, , . ¹⁴- in vitro ( ) , in vivo, . .     

Mass spectrometric determination of the amino acid sequence in arginine-containing peptides

aa, a-a a- a aa - aa a -a .     

热压烧结ZrB2／B4C陶瓷材料的组织与性能

以B4C、C、ZrO2为主要原料,采用反应热压法制备ZrB2／B4C陶瓷材料。通过对ZrB2／B4C陶瓷材料的显微组织及力学性能的综合分析,发现第二相ZrB2含量为20wt％时,材料具有较好的综合力学性能,相对密度为99．3％,维氏硬度为36．1GPa,抗弯强度为533．3MPa,断裂韧性为6．95MPa·m^1/2,比纯B4C陶瓷材料的性能均有所提高。材料为穿晶和沿晶断裂的混合断裂模式。     

Gender- and site-related effects on lipolytic capacity of rat white adipose tissue

Gender- and site-related differences in the lipolytic capacity, at the different steps of the adrenergic pathway, in gonadal and inguinal white adipose tissue (WAT), were assessed by studying _2A-adrenergic receptor (AR), ₃-AR and hormone-sensitive lipase (HSL) protein levels, and by determining the lipolytic response to different agents. Gonadal WAT showed a lower _2A/₃-AR ratio, a greater lipolytic capacity in response to AR agonists, and higher HSL activity and protein levels than inguinal WAT. In female rats, we found greater _2A-AR protein levels and _2A/₃-AR ratio compared to their male counterparts, but, on the other hand, a higher lipolytic response to -AR agonists and a greater lipolytic capacity at the postreceptor level, including a more activated HSL protein. Thus, the lipolytic capacity was clearly higher in gonadal than in inguinal WAT, at the different steps of the adrenergic pathway studied. Moreover, in both tissues, females showed a greater inhibition of lipolysis via ₂-AR, which was counteracted by the higher lipolytic capacity at the postreceptor level.Received 1 April 2003; received after revision 11 June 2003; accepted 23 June 2003     

P16及COX-2蛋白在宫颈癌中表达及意义

目的 探讨P16及COX-2蛋白在宫颈癌中表达及意义.方法 收集宫颈活检或手术标本138例,采用免疫组织化学方法分别检测P16及COX-2蛋白的表达,并分析结果.结果 正常宫颈组织、宫颈低级别上皮内瘤变(LSIL)、宫颈高级别上皮内瘤变(HSIL)、宫颈癌中P16蛋白阳性表达分别为0％(0/30)、42.3％ (11/26)、71.9％(23/32)、92％(46/50).COX-2蛋白阳性表达分别为3.3％(1/30)、50％(13/26)、75％(24/32)、88％ (44/50).随着宫颈病变分级升高,P16及COX-2蛋白阳性率逐渐上升,统计学差异有显著性(p＜0.05);结论 P16及COX-2的高表达在宫颈癌的发生、发展中均起作用.     

Photoreactivation of UV damage in cultured Drosophila cells

Cell survival and photoreactivation of 254 nm ultraviolet (UV) light damage in a wild type Drosophila cell line was assayed by colony formation in liquid medium. Fo, Fq, and extrapolation number for the exponential portion of survival curves are 21 J/m2, 3.6 J/m2, and 1.5 for non-photoreactivated cells and 110 J/m2, 11.2 J/m2, and 1.3 for those exposed to photoreactivating light. Maximal photoreactivation occurs at the 100 J/m2 region of the curve. At 10 and 50% survival, 75-80% of the UV damage was photoreactivable.     

The product of the γ-secretase processing of ephrinB2 regulates VE-cadherin complexes and angiogenesis

Presenilin-1 (PS1) gene encodes the catalytic component of γ-secretase, which proteolytically processes several type I transmembrane proteins. We here present evidence that the cytosolic peptide efnB2/CTF2 produced by the PS1/γ-secretase cleavage of efnB2 ligand promotes EphB4 receptor-dependent angiogenesis in vitro. EfnB2/CTF2 increases endothelial cell sprouting and tube formation, stimulates the formation of angiogenic complexes that include VE-cadherin, Raf-1 and Rok-α, and increases MLC2 phosphorylation. These functions are mediated by the PDZ-binding domain of efnB2. Acute downregulation of PS1 or inhibition of γ-secretase inhibits the angiogenic functions of EphB4 while absence of PS1 decreases the VE-cadherin angiogenic complexes of mouse brain. Our data reveal a mechanism by which PS1/γ-secretase regulates efnB2/EphB4 mediated angiogenesis.     

MAPK/ERK1/2 signaling mediates endothelial-like differentiation of immature DCs in the microenvironment of esophageal squamous cell carcinoma

Endothelial-like differentiation of dendritic cells (DCs) is a new phenomenon, and the mechanism is still elusive. Here, we show that the tumor microenvironment derived from the human esophageal squamous cell carcinoma (ESCC) cell line EC9706 can induce immature DCs (iDCs) differentiate toward endothelial cells, and become endothelial-like cells, but it has no obvious influence on mature DCs. During the course of endothelial-like differentiation of iDCs, a sustained activation of mitogen-activated protein kinase/extracelluar signal-regulated kinase1/2 (MAPK/ERK1/2) and cAMP response element-binding protein (CREB) was detected. Incubation of iDCs with MEK phosphorylation inhibitor PD98059 blocked the MAPK/ERK1/2 and CREB phosphorylation as well as the endothelial-like differentiation of iDCs. Inhibition of vascular endothelial growth factor-A (VEGF-A) in the microenvironment with its antibody blocked the endothelial-like differentiation and the phosphorylation of MAPK/ERK1/2 and CREB. These data suggest that MAPK/ERK1/2 signaling pathway activated by VEGF-A could mediate endothelial-like differentiation of iDCs in the ESCC microenvironment.     

Proton-induced dismutation of superoxide in aprotic media by bile pigments

Summary Cyclic voltammetry of molecular oxygen in aprotic media (dimethylformamide) and in the presence of bilirubin and other bile pigments shows that superoxide anion ( ) undergoes proton-induced dismutation. Lactam hydrogens of bile pigments are sufficiently acid to induce disproportionation to O₂ and H₂O₂. Because of its characteristic lipophilic behavior, a biological role for natural bilirubin similar to that of other non-enzymatic lipophilic scavengers of is suggested.     

Oxygen consumption in human placental lobules in vitro (survival maintained by perfusion)]

The average oxygen utilization coefficient of 26% is calculated from PO2 determinations during the perfusion of the fetal circulation of isolated human term placental lobules. The oxygen consumption approximates 0.4 ml/min/kg placenta when Earle's salt solution is used as a perfusion medium and is equilibrated with a gas mixture containing 5% O2. The oxygen consumption reaches 2 ml/min/kg when the same Earle's solution is equilibrated with a gas mixture containing 95% O2.     

Induction of lymphomas by urethane in combination with diethylstilboestrol in CFLP mice

Summary The combination of urethane and the non-steroidal oestrogen diethylstilboestrol induced lymphomas in CFLP mice.This work was supported by grants from the Ministry of Health of Hungary (Grants No. 2-10-0401-01-2/K, 2-11-0801-01-1/VR, 2-09-0801-01-1/E).     

In vivo effects of immunostimulating lipopeptides on mouse liver microsomal cytochromes P-450 and on paracetamol-induced toxicity

Summary Immunomodulating lipopeptides lauroyl-L-Ala--D-Glu-LL-A2pmNH2-Gly (RP 44.102) and lauroyl-L-Ala--D-Glu-LL-A2pmNH2 (RP 56.142) were found to protect mice against the hepatotoxicity of paracetamol, which is due to cytochrome P-450 dependent formation of toxic metabolites and radicals. In fact they decreased the amount of hepatic microsomal cytochrome P-450, and the level of CCl₄-induced lipid peroxidation. In contrast lauroyl-L-Ala--D-Glu-DD-A2pmNH2 (RP 53.204), which only differs by the configuration of the two chiral carbons of A2pm (diaminopimelic acid) and is not an immunomodulating agent, failed to protect against poisoning by paracetamol and had no effect on the level of hepatic cytochrome P-450 or the microsomal CCl₄-induced lipid peroxidation. This provides a clear connection between the immunostimulating properties of a compound and its effects on xenobiotic biotransformations.     

Regulation of macrophage activation

IFN- rapidly primes the macrophage via JAK1/2-STAT1 pathway so that it can subsequently undergo a slower classical type 1 activation upon exposure to T helper (Th)1 cytokines such as IFN or other activators, including tumor necrosis factor and lipopolysaccharide, e.g. in intracellular killing of phagocytosed Mycobacterium tuberculosis. If instead it is driven by Th2 cytokines interleukin (IL)-4 and IL-13, it undergoes alternate type 2 activation, which enhances endocytotic antigen uptake and presentation, mast cell and eosinophil involvement and type 2 granuloma formation, e.g. in response to parasitic and extracellular pathogens. Particle-induced macrophage activation was shown to differ from classical and alternate activation, showing in DNA microarray experiments (complete linkage/ Euclidean distance metric analysis) upregulation of nonsecreted structural/signaling molecules and lack of secreted proin-flammatory cyto- and chemokines. The switch-off (deactivation) of already activated macrophages is an active, controlled process in which IL-10 and corticosteroids play important roles and to which15dPGJ2, PGA1/2 and vasoactive intestinal peptide often contribute.Received 16 January 2003; received after revision 14 March 2003; accepted 2 April 2003