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Journal of Biosciences and Medicines 2016年第4期4卷 68-79页

作者： Bronislaw L. Slomiany amalia Slomiany Research Center Rutgers School of Dental Medicine Rutgers The State University of New Jersey Newark NJ USA

Matrix metalloproteinase-9 (MMP-9) is a highly glycosylated endopeptidase implicated in a wide rage of oral mucosal inflammatory and neoplastic diseases, including chronic periodontitis, a persistent mucosal inflammation attributed primarily to infection by oral anaerobe, P. gingivalis. In this study, we explored the role of Rac1 and mitogen-activated protein kinases (MAPKs) in the processes of MMP-9 release in sublingual salivary gland cells exposed to P. gingivalis key endotoxin, cell wall lipopolysaccharide (LPS). We demonstrate that the LPS-elicited induction in the acinar cell MMP-9 release is associated with MAPK, ERK and p38 activation, and occurs with the involvement of Rac1 and cytosolic phospholipase A2 (cPLA2). Further, we reveal that the LPS-induced MMP-9 release involves ERK-mediated phosphorylation of cPLA2 on Ser505 that is essential for its membrane translocation with Rac1, and that this process requires p38 activation. Moreover, we show that phosphorylation and membrane localization of p38 with Rac1-GTP play a pivotal role in cPLA2-dependent induction in MMP-9 release. Thus collectively, our findings infer that P. gingivalis LPS-induced up-regulation in the acinar cell MMP-9 release requires ERK-dependent recruitment of cPLA2 to the membrane localized Rac1/p38 complex.

关键词： P. gingivalis LPS Oral Mucosa Rac1 p38 ERK cPLA2 Activation MMP-9 Release

New roles of poison control centres

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New roles of poison control centres

Anti-VDAC3 recombinant antibody decreased human sperm motility and membrane integrity: A potential spermicide for contraception

The 5th Congress of Toxicology in Developing Countries

作者： amalia Laborde Centro de Informacin y Asesoramiento Toxicolgico Department of Toxicology Faculty of Medicine Montevideo Uruguay

＜正＞ The primary mission of poison control centres has always been the improvement of poisoned patient care and poison prevention, but the needs for achieving this mission imply many functions and roles to be accomplished. Many centres, even in developing countries, are multifunctional and provide broad toxicological information services. The main challenges of poison centres in developing countries

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Asian pacific Journal of Reproduction 2017年第6期6卷 257-263页

作者： Asmarinah Tri Panjiasih Susmiarsih amalia Shari Putri Ratri Dwi Ari Pujianto Endang Winiati Bachtiar Department of Medical Biology Faculty of Medicine University of Indonesia Postgraduate Program in Biomedical Sciences Faculty of MedicineUniversity of Indonesia Faculty of Medicine YARSI UniversityJakartaIndonesia Department of Oral Biology Faculty of Dentistry University of Indonesia

Objective:To express recombinant protein that comprises an important fragment of human sperm specific voltage dependent anion channel 3 (VDAC3) protein as a potential molecule for generation of antibody, which can affect sperm function, aiming at spermicide development. Methods: The produce of VDAC3 recombinant protein encoded by cDNA sequence of human VDAC3 exon 5-8, based on experimental design of VDAC3 knock-out mice study. And after the purification of various human sperm VDAC3 recombinant proteins, epitope has been predicted in our recombinant protein determined by ElliPro program. Polyclonal antibody was produced for 14 wk. Then anti-VDAC3-exon 5-8 recombinant antiserum was inoculated to human sperm. After the process, antibody VDAC3 protein in human sperm was incubation with anti-VDAC3 recombinant antibody. Finally evaluation the effect of VDAC3 antiserum to human sperm motility and plasma membrane integrity was ***: Human VDAC3 recombinant protein was successfully over-expressed in Escherichia coli and purified by affinity chromatography method. Purified human sperm VDAC3 recombinant protein could stimulate immune response in rabbit producing an antibody against VDAC3. Anti-VDAC3 recombinant antibody recognized VDAC3 antigen in human sperm could decrease human sperm motility and membrane integrity ***:Anti-VDAC3 recombinant polyclonal antibody that we produced in rabbit by ourselves could decrease sperm motility and sperm membrane integrity. The authors suggest this polyclonal antibody could be used as a candidate agent for male contraception in the future. Furthermore, the authors intend to explore the effect of this antibody into sperm function aiming at male contraceptive vaccine development.

关键词： Recombinant VDAC3 Antibody Sperm motility Membrane integrity Spermicide

Role of α-Tubulin Acetylation and Protein Kinase D2 Ser/Tyr Phosphorylation in Modulation by Ghrelin of Porphyromonas gingivalis-Induced Enhancement in Matrix Metalloproteinase-9 (MMP-9) Secretion by Salivary Gland Cells

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Journal of Biosciences and Medicines 2016年第7期4卷 82-94页

作者： Bronislaw L. Slomiany amalia Slomiany Research Center Rutgers School of Dental Medicine Rutgers The State University of New Jersey Newark NJ USA

Matrix metalloproteinas-9 (MMP-9) is a glycosylated endopeptidase, and hence its processing between the endoplasmic reticulum (ER), Golgi and trans-Golgi (TGN) network remains under a strict control of factors that affect the microtubule (MT) stabilization, and the recruitment and activation of coat and cargo proteins, including ADP-ribosylation factors (Arfs) and protein kinase D (PKD). Here, we report on the factors implicated in the regulation of MMP-9 secretion by salivary gland acinar cells in response to P. gingivalis LPS, and the effect of hormone, ghrelin. We show that the LPS-elicited induction in MMP-9 secretion is associated with the increase in α-tubulin acetylation and the enhancement in MT stabilization, while the modulatory effect of ghrelin is reflected in a decrease in α-tubulin acetylation. Further, the effect of the LPS occurs in concert with up-regulation in Arf-guanine nucleotide exchange factor (GEF)-mediated Arf1 activation and the TGN recruitment of PKD2, while ghrelin exerts the modulatory effect on Arf-GEF activation. Moreover, we reveal that the LPS-induced up-regulation in MMP-9 secretion is reflected in a marked increase in PKCδ-mediated PKD2 phosphorylation on Ser, while the modulatory effect of ghrelin is manifested by the SFK-PTKs-dependent phosphorylation of PKD2 on Tyr. The findings demonstrate that MT stabilization along with Arf-GEF-mediated Arf1/PKD2 activation play a major role in P. gingivalis LPS-induced up-regulation in salivary gland acinar cell MMP-9 secretion, and point the modulatory mode of action by ghrelin.

关键词： Porphyromonas gingivalis Oral Mucosa Ghrelin MMP-9 α-Tubulin Acetylation Arf1 PKD2 Ser/Tyr Phosphorylation

全新世东地中海的陆源碎屑输入及其水文气候变化:海盆尺度地球化学分析

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地学前缘 2022年第4期29卷 156-167页

作者：吴家望姚胜男 amalia FILIPPIDI 刘志飞 Gert JDE LANGE 中山大学海洋科学学院广东珠海519082 乌特勒支大学地球科学系荷兰乌特勒支3508TA 同济大学海洋地质国家重点实验室上海200092

地中海位于非洲季风气候和欧洲温带气候的交界处,同时接受周边地区岩石性质和风化状况差异极大的碎屑物质,所以是研究地球表层水文循环的理想区域。前人的认识集中于撒哈拉风尘和尼罗河输入,往往忽视了其他的陆源碎屑沉积,尤其对不同水... 详细信息

地中海位于非洲季风气候和欧洲温带气候的交界处,同时接受周边地区岩石性质和风化状况差异极大的碎屑物质,所以是研究地球表层水文循环的理想区域。前人的认识集中于撒哈拉风尘和尼罗河输入,往往忽视了其他的陆源碎屑沉积,尤其对不同水文气候条件下的潜在变化缺乏考虑。针对形成于全新世非洲湿润期的腐泥层S1页岩沉积,并结合采自岩心顶部的晚全新世/现代沉积物,文章从18个站位选取了30个样品开展碎屑组分的地球化学分析,通过在具有不同干湿环境背景的时间片段上开展盆地尺度的对比(约9.5~8.9 ka vs.约1.7~0 ka),探讨全新世东地中海的陆源碎屑输入模式。Ti/Al、Zr/Al、Ca/Al、Y/Al都清晰显示了经向和纬向上的梯度变化,可用作撒哈拉风尘的可靠指标;这些碎屑元素之间的差异反映了北非风尘来源和传送路径的变化。这些风尘指标在南-北向上的一致变化指示了副热带高压与西风带的交互界限为36°N,该界限在全新世应稳定存在。与风尘相反,河流输入的指标值在早全新世时显著高于现代,并呈现不同的地理分布。K/Al具有西高东低、北高南低的分布特征,指示了地中海北部沿岸地区的河流输入。根据与Ti/Al、K/Al的差异,利用(Cr+Ni)/Al可较好地识别被增强的季风降雨所激活的北非古河流沉积。河流输入在早全新世时显著上升,但不同河流系统的影响范围差异很大,受制于源区水文气候、表层洋流搬运等因素。

关键词：地中海全新世非洲湿润期腐泥层S1 元素地球化学撒哈拉风尘古河流

Modulation of gastric mucosal inflammatory responses to Helicobacter pyloriby ghrelin: Role of cNOS-dependent IKK-βS-nitrosylation in the regulation of COX-2 activation

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American Journal of Molecular Biology 2012年第2期2卷 113-123页

作者： Bronislaw L. Slomiany amalia Slomiany Research Center University of Medicine and Dentistry of New Jersey Newark USA

Disturbances in nitric oxide synthase (NOS) and cyclooxygenase (COX) isozyme systems, manifested by the excessive NO and prostaglandin (PGE2) generation, are well-recognized features of gastric mucosal inflammatory responses to H. pylori infection. In this study, we report that H. pylori LPS-induced enhancement in gastric mucosal inducible (i) iNOS expression and COX-2 activation was accompanied by the impairment in constitutive (c) cNOS phosphorylation, up-regulation in the inhibitory κB kinase-β (IKKβ) activation and the increase in the transcriptional factor, NF-κB, nuclear translocation. Further, we show that abrogation of cNOS control over NF-κB activation has lead to induction of iNOS expression and COX-2 activation through S-nitrosylation. Moreover, we demonstrate that the modulatory effect of peptide hormone, ghrelin, on the LPS-induced changes was reflected in the increase in Src/Akt-dependent cNOS activation through phosphorylation and the suppression of IKK-β activity through cNOS-mediated IKK-β protein S-nitrosylation. As a result, ghrelin exerted the inhibitory effect on NF-κB nuclear translocation, thus causing the repression of iNOS gene induction and the inhibition in COX-2 activation through iNOS-dependent S-nitrosylation. Our findings point to cNOS activation as a pivotal element in the signaling cascade by which ghrelin exerts modulatory control over proinflammatory events triggered in gastric mucosa by H. pylori infection.

关键词： H. pylori Gastric Mucosa iNOS Induction COX-2 Activation Ghrelin cNOS Phosphorylation IKK-β S-Nitrosylation

Ghrelin-induced cSrc activation through constitutive nitric oxide synthase-dependent S-nitrosylation in modulation of salivary gland acinar cell inflammatory responses to Porphyromonas gingivalis

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American Journal of Molecular Biology 2011年第2期1卷 43-51页

作者： Bronislaw L. Slomiany amalia Slomiany 不详

A peptide hormone, ghrelin, recognized for its role in the regulation of nitric oxide production has emerged as an important modulator of oral mucosal inflammatory responses to periodontopathic bacterium, P. gingivalis. As cSrc kinase plays a major role in controlling the activity of nitric oxide synthase (NOS) system, in this study we investigated the influence of P. gingivalis LPS on the processes of Src activation in rat sublingual gland acinar cells. The LPS-induced enhancement in the activity of inducible (i) iNOS and the impairment in constitutive (c) cNOS were reflected in the suppression in cSrc activity and the extent of its phosphorylation at Tyr416. Further, we show that the countering effect of ghrelin on the LPS-induced changes in cSrc activity and the extent of its phosphorylation was accompanied by a marked reduction in iNOS and the increase in cNOS activation through phosphorylation at Ser1179. Moreover, the effect of ghrelin on cSrc activation was associated with the kinase S-nitrosylation that was susceptible to the blockage by cNOS inhibition. Our findings suggest that P. gingivalis-induced up-regulation in iNOS leads to disturbances in cNOS phosphorylation that exerts the detrimental effect on the processes of cSrc activation through cNOS mediated S-nitrosylation. We also show that the effect of ghrelin on P. gingivalis-induced inflammatory changes are manifested in the enhancement in cSrc activation through S-nitrosylation and the increase in its phosphorylation at Tyr416.

关键词： Ghrelin P. Gingivalis Salivary Acinar Cells cNOS cSrc Activation S-Nitrosylation

Porphyromonas gingivalis-Induced GEF Dock180 Activation by Src/PKCδ-Dependent Phosphorylation Mediates PLCγ2 Amplification in Salivary Gland Acinar Cells: Modulatory Effect of Ghrelin

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Journal of Biosciences and Medicines 2015年第7期3卷 66-77页

作者： Bronislaw L. Slomiany amalia Slomiany Research Center Rutgers School of Dental Medicine Rutgers The State University of New Jersey Newark NJ USA

Phospholipase Cγ2 (PLCγ2) plays a pivotal role in mediation of inflammatory reaction to bacterial lipopolysaccharide (LPS) as well as serves as a key target in modulatory influence of the hormone ghrelin. Here we explore the involvement of Rac1 and its activator, guanine nucleotide exchange factor (GEF), Dock180, in mediation of PLCγ2 activation in salivary gland acinar cells in response to P. gingivalis LPS and ghrelin. We show that stimulation of the acinar cells with the LPS leads to up-regulation in Dock and PLCγ2 activation, and is reflected in the membrane translocation of Rac1 and PLCγ2, while the effect of ghrelin is manifested by the suppression in Rac1 translocation. Further, we reveal that stimulation with the LPS leads to Dock180 phosphorylation on Tyr and Ser, while the modulatory influence of ghrelin, manifested by a drop in membrane Rac1-GTP, is asso-ciated with a distinct decrease in Dock180 phosphorylation on Ser. Moreover, we demonstrate that phosphorylation on Tyr remains under the control of Src kinase and is accompanied by Dock180 membrane translocation, while protein kinase Cδ(PKCδ) is involved in the LPS-induced phosphorylation of the membrane-recruited Dock180 on Ser. Thus, our findings underscore the role of Src/PKCδ-mediated GEF Dock180 phosphorylation on Tyr/Ser in modulation of salivary gland acinar cell PLCγ2 activation in response to P. gingivalis as well as ghrelin.

关键词： P. gingivalis Salivary Acinar Cell Ghrelin Dock180 Phosphorylation Src PKCδ PLCγ2 Activation

Cytosolic phospholipase A2 S-nitrosylation in ghrelin protection against detrimental effect of ethanol cytotoxicity on gastric mucin synthesis ——Ghrelin in gastric mucosal protection

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Health 2010年第9期2卷 1033-1039页

作者： Bronislaw L. Slomiany amalia Slomiany 不详

Ghrelin, a peptide hormone produced mainly in the stomach, has emerged recently as an important regulator of nitric oxide synthase (NOS) and cyclooxygenase (COX) enzyme systems, the products of which play direct cytoprotective function in the maintenance of gastric mucosal integrity. In this study, using gastric mucosal cells, we report on the role of ghrelin in countering the cytotoxic effect of ethanol on mucin synthesis. We show that the countering effect of ghrelin on mucin synthesis was associated with the increase in NO and PGE2 production, and characterized by a marked up-regulation in cytosolic phospholipase A2 (cPLA2) activity. The ghrelin-induced up-regulation in mucin synthesis, like that of cPLA2 activity, was subject to suppression by Src inhibitor, PP2 and ERK inhibitor, PD98059, as well as ascorbate. Moreover, the loss in countering effect of ghrelin on the ethanol cytotoxicity and mucin synthesis was attained with cNOS inhibitor, L-NAME as well as COX-1 inhibitor SC-560. The effect of L-NAME was reflected in the inhibition of ghrelin-induced mucosal cell capacity for NO production, cPLA2 S-nitrosylation and PGE2 generation, whereas COX-1 inhibitor caused only the inhibition in PGE2 generation. Our findings suggest that the activation of gastric mucosal cPLA2 through cNOS-induced S-nitrosylation plays an essential role in the countering effect of ghrelin on the disturbances in gastric mucin synthesis caused by ethanol cytotoxicity.

关键词： Ghrelin Ethanol Cytotoxicity Gastric Mucin CNOS CPLA2 S-Nitrosylation

Role of Protein Kinase Cδ-Mediated Spleen Tyrosine Kinase (Syk) Phosphorylation on Ser in the Amplification of Oral Mucosal Inflammatory Responses to Porphyromonas gingivalis

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Journal of Biosciences and Medicines 2018年第3期6卷 70-85页

作者： Bronislaw L.Slomiany amalia Slomiany Research Center Rutgers School of Dental MedicineRutgersThe State University of New JerseyNewarkNJUSA

The signaling events underlying oral mucosal inflammatory responses to P. gingivalis and its key endotoxin, lipopolysaccharide (LPS), relay primarily on the LPS engagement of Toll-like receptor-4 (TLR4), and the activation of IκB-kinase complex (IKK) and mitogen-activated protein kinases (MAPKs that exert their control over transcription factors implicated in the regulation of iNOS and COX-2 proinflammatory genes expression). Since spleen tyrosine kinase (Syk) has emerged recently as a major amplifier in the production of proinflammatory mediators, we investigated the process of recruitment and interaction of Syk with TLR4 in salivary gland acinar cells in response to P. gingivalis LPS. Our findings revealed that stimulation of the acinar cells with the LPS leads to protein kinase Cδ (PKCδ)-mediated phosphorylation of Syk on Ser which results in its localization with the membrane associated TLR4 complex and the activation through phosphorylation on Tyr. Further, our results support the involvement of Syk in the amplification of transcription factors involved in the assembly and expression of transcription complexes associated with the induction in COX-2 and iNOS genes. Therefore, our data suggest that PKCδ is a primary linchpin affecting the Syk recruitment to the membrane localized TLR4, and hence affects the efficiency of the kinase activation and the magnitude of oral mucosal inflammatory response to P. gingivalis.

关键词： P. gingivalis Oral Mucosa PKCδ Syk Activation Ser/Tyr Phosphorylation