Antimicrobial effects of terpinen-4-ol against oral pathogens and its capacity for the modulation of gene expression

Abstract

This study evaluated the antibacterial activity of terpinen-4-ol against Streptococcus mutans and Lactobacillus acidophilus and its influence on gbpA (S. mutans) and slpA (L. acidophilus) gene expression. As measured by XTT assay, the concentrations of terpinen-4-ol that effectively inhibited the biofilm were 0.24% and 0.95% for S. mutans and L. acidophilus, respectively. Confocal microscopy revealed the presence of a biofilm attached to the enamel and dentin block surfaces with significant terpinen-4-ol effects against these microorganisms. The expression of the gbpA and slpA genes involved in adherence and biofilm formation was investigated using RT-PCR. Expression of these genes decreased after 15?min with 0.24% and 0.95% terpinen-4-ol in S. mutans and L. acidophilus, respectively. These findings demonstrate the antimicrobial activity of terpinen-4-ol and its ability to modulate the expression of gbpA and slpA genes, emphasizing the therapeutic capacity of terpinen-4-ol as an alternative to inhibit adherence in biofilm.     

杂拟谷盗的热适应特性

为了解杂拟谷盗Tribolium confusum Jacquelin du Val的热适应特性,将杂拟谷盗分别于15,25和35℃下驯化2周后,用温度梯度仪测量在不同温度驯化下杂拟谷盗的最适温度、临界低温和临界高温。结果表明,驯化温度对杂拟谷盗最适温度、临界低温和临界高温的影响极显著(P<0.01),最适温度、临界低温、临界高温均随着驯化温度的升高而升高。最适温区的范围随着驯化温度的升高而扩大。驯化温度对杂拟谷盗最适温度的影响最大(0.317),对临界低温的影响(0.310)大于临界高温(0.255)。     

In vitro antiviral activity of Melaleuca alternifolia essential oil

Aims:  To investigate the in vitro antiviral activity of Melaleuca alternifolia essential oil (TTO) and its main components, terpinen-4-ol, α-terpinene, γ-terpinene, p -cymene, terpinolene and α-terpineol.
Methods and Results:  The antiviral activity of tested compounds was evaluated against polio type 1, ECHO 9, Coxsackie B1, adeno type 2, herpes simplex (HSV) type 1 and 2 viruses by 50% plaque reduction assay. The anti-influenza virus assay was based on the inhibition of the virus-induced cytopathogenicity. Results obtained from our screening demonstrated that the TTO and some of its components (the terpinen-4-ol, the terpinolene, the α-terpineol) have an inhibitory effect on influenza A/PR/8 virus subtype H1N1 replication at doses below the cytotoxic dose. The ID₅₀ value of the TTO was found to be 0·0006% (v/v) and was much lower than its CD₅₀ (0·025% v/v). All the compounds were ineffective against polio 1, adeno 2, ECHO 9, Coxsackie B1, HSV-1 and HSV-2. None of the tested compounds showed virucidal activity. Only a slight virucidal effect was observed for TTO (0·125% v/v) against HSV-1 and HSV-2.
Conclusions:  These data show that TTO has an antiviral activity against influenza A/PR/8 virus subtype H1N1 and that antiviral activity has been principally attributed to terpinen-4-ol, the main active component.
Significance and Impact of the Study:  TTO should be a promising drug in the treatment of influenza virus infection.     

Comparison of the cidal activity of tea tree oil and terpinen-4-ol against clinical bacterial skin isolates and human fibroblast cells

Aims: The aim of this study was to compare both the antimicrobial activity of terpinen‐4‐ol and tea tree oil (TTO) against clinical skin isolates of meticillin‐resistant Staphylococcus aureus (MRSA) and coagulase‐negative staphylococci (CoNS) and their toxicity against human fibroblast cells. Methods and Results: Antimicrobial activity was compared by using broth microdilution and quantitative in vitro time‐kill test methods. Terpinen‐4‐ol exhibited significantly greater bacteriostatic and bactericidal activity, as measured by minimum inhibitory and bactericidal concentrations, respectively, than TTO against both MRSA and CoNS isolates. Although not statistically significant, time‐kill studies also clearly showed that terpinen‐4‐ol exhibited greater antimicrobial activity than TTO. Comparison of the toxicity of terpinen‐4‐ol and TTO against human fibroblasts revealed that neither agent, at the concentrations tested, were toxic over the 24‐h test period. Conclusions: Terpinen‐4‐ol is a more potent antibacterial agent against MRSA and CoNS isolates than TTO with neither agent exhibiting toxicity to fibroblast cells at the concentrations tested. Significance and Impact of the Study: Terpinen‐4‐ol should be considered for inclusion as a single agent in products formulated for topical treatment of MRSA infection. However, further work would initially be required to ensure that resistance would not develop with the use of terpinen‐4‐ol as a single agent.     

Antifungal activity of the components of Melaleuca alternifolia (tea tree) oil

AIMS: To investigate the in vitro antifungal activity of the components of Melaleuca alternifolia (tea tree) oil. METHODS AND RESULTS: Activity was investigated by broth microdilution and macrodilution, and time kill methods. Components showing the most activity, with minimum inhibitory concentrations and minimum fungicidal concentrations of < or =0.25%, were terpinen-4-ol, alpha-terpineol, linalool, alpha-pinene and beta-pinene, followed by 1,8-cineole. The remaining components showed slightly less activity and had values ranging from 0.5 to 2%, with the exception of beta-myrcene which showed no detectable activity. Susceptibility data generated for several of the least water-soluble components were two or more dilutions lower by macrodilution, compared with microdilution. CONCLUSIONS: All tea tree oil components, except beta-myrcene, had antifungal activity. The lack of activity reported for some components by microdilution may be due to these components becoming absorbed into the polystyrene of the microtitre tray. This indicates that plastics are unsuitable as assay vessels for tests with these or similar components. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has identified that most components of tea tree oil have activity against a range of fungi. However, the measurement of antifungal activity may be significantly influenced by the test method.     

不同杀虫剂处理对赤拟谷盗P450基因诱导表达特性影响

害虫细胞色素P450基因可被杀虫剂迅速诱导,然而当前对不同杀虫剂处理下赤拟谷盗P450基因诱导表达特性的研究较少。本研究首先通过序列比对选取了来自不同家族的8个赤拟谷盗P450基因CYP4G7、CYP4Q4、CYP4BR3、CYP12H1、CYP6BK11、CYP9D4、CYP9Z5和CYP345A1,然后采用四种不同杀虫剂氯氰菊酯、氟氯氰菊酯、氯菊酯和吡虫啉对赤拟谷盗20 d幼虫进行生物测定,再根据生测结果以四个药剂亚致死剂量分别处理幼虫,并采用荧光定量PCR分析8个P450基因的表达特性。结果表明,CYP4G7和CYP345A1可以分别被氯氰菊酯(分别上调1.97倍和2.06倍)、氟氯氰菊酯(2.00倍和2.03倍)和氯菊酯(1.73倍和1.81倍)显著诱导,而CYP4BR3和CYP345A1可以被吡虫啉(分别上调1.99倍和1.93倍)显著诱导。本研究结果表明赤拟谷盗P450基因的显著诱导与基因家族类型以及农药品种有关。     

赤拟谷盗和杂拟谷盗的生殖隔离研究

赤拟谷盗和杂拟谷盗是两个同域发生的近缘储粮害虫。为明确它们的生殖隔离程度和机制,本研究比较分析了赤拟谷盗和杂拟谷盗雄虫对同种和异种雌虫的交配选择;赤拟谷盗与杂拟谷盗交配后,测定了精子在异种雌体内的存活情况;将赤拟谷盗和杂拟谷盗进行正反杂交,研究其F1代、F2代和回交代杂种是否产生及其雌雄比。结果显示,赤拟谷盗和杂拟谷盗雄虫对同种雌虫的爬跨率和交配率高于其对异种雌虫的爬跨率和交配率,表明赤拟谷盗和杂拟谷盗种间性隔离不完全;赤拟谷盗和杂拟谷盗交配后精子在异种雌体内是存活的,杂交所产的F1代卵为受精卵,说明交配后完成了精子传送和受精过程,表明种间机械隔离和配子隔离不完全;赤拟谷盗和杂拟谷盗的杂种F1代自交和回交产生了F2代和回交代,表明种间不存在杂种不育;一些杂交组合产生的F2代和回交代数量少且存在雌雄性比偏离,表明种间存在部分杂种不活和杂种衰败。本研究明确了赤拟谷盗和杂拟谷盗的生殖隔离机制,这对于揭示它们的物种进化关系有着重要意义。     

松油烯-4-醇对粘虫的致毒机制

采用华氏呼吸仪法、常规生化酶活力测定等方法,测定松油烯-4-醇熏蒸处理对粘虫Mythimna separata(Walker)幼虫呼吸作用、血淋巴理化性状、体内酶系活性等的影响。结果表明,松油烯-4-醇显著地影响粘虫的呼吸作用,不同中毒阶段试虫的呼吸率均显著提高,呼吸商发生改变;血淋巴理化性状也发生一定程度的变化,血淋巴总量随中毒程度的加深呈下降趋势,兴奋期、痉挛期、昏迷期血淋巴总量分别为对照试虫的85.55%、70.39%、38.47%,血淋巴比重呈上升趋势,pH值和渗透压变化不大;体内Na+-K+-ATP酶的活性受到明显抑制,头部组织中Na+-K+-ATP酶活性的抑制率在兴奋期、痉挛期分别达36.4%、80.2%,中肠组织中Na+-K+-ATP酶活性的抑制率达50%左右,对乙酰胆碱脂酶活性影响不大,对酯酶则是先激活后抑制。松油烯-4-醇对Na+-K+-ATP酶活性的抑制可能与粘虫最终死亡有关。     

Cleavage of α-Dicarbonyl Compounds by Terpene Hydroperoxide

Thermostabilities of component enzymes in the pyruvate dehydrogenase complex from Bacillus stearothermophilus decreased in the order lipoamide dehydrogenase, lipoate acetyltransferase, and pyruvate decarboxylase (E1). Fluorescence of an extrinsic 8-amino-1-naphthalenesulfonate (ANS) increased with inactivation of E1. The thermal denaturation of the enzymes resulted in disassembly of the complex. El was involved in a resulting aggregate of the complex. The interaction between ANS and denatured E1 accounted for an increase in fluorescence.     

杂拟谷盗体内 Wolbachia 感染密度的时间和空间分布状况

为搞清杂拟谷盗体内沃尔巴克氏体(Wolbachia)感染密度的时间和空间分布状况,本研究采用实时荧光定量PCR方法测定了杂拟谷盗不同发育阶段、不同日龄、不同性别和不同身体部位的Wolbachia感染密度。结果表明,杂拟谷盗在卵和成虫期Wolbachia感染密度高于其幼虫和蛹期,成虫腹部的Wolbachia感染密度高于其头部和胸部,而成虫不同日龄和雌雄之间的Wolbachia感染密度均没有显著性差异。本研究明确了Wolbachia在杂拟谷盗体内的时空分布和动态变化规律,这对于揭示寄主与共生菌之间的互作关系有着重要意义。     

The Influence of Oxidizing Agents on Macromolecular Synthesis for Chitinous Material

Summary

The 5th instar larvae of the common flour beetle, Tribolium confusum, was utilized as an invertebrate model to test the larvicidal potential of oxidizing agents used as flour preservatives. Larvae were exposed to 10 ppm of either diflubenzuron or benzoyl peroxide. Results showed decreases in both total protein and glucose concentrations. Histological examination also indicated smaller deposition of chitinous material in the endocuticle. Both treatments had similar effects. Thus, these data indicate that oxidizing agents mixed in flour may be used for the control of development of flour beetle larvae.     

烟草叶片衰老期过程中的蛋白质组学分析

大田烟叶生产过程中因打顶打叉的处理,改变了烟叶正常的衰老模式。为研究这一特殊的衰老机制,我们自旺长期开始,对‘云烟87’不同发育阶段烟株的中部叶片,进行形态观测、生理生化分析及蛋白质组学检测。结果显示：随着烟叶的逐渐成熟和衰老,烟草的叶色逐渐变黄,叶片逐渐变短、变窄,厚度减少;解剖结构清晰看到栅栏组织和海绵组织从最初的整齐排列到逐渐排列紊乱,组织细胞间轮廓不明显,细胞间隙明显增大;亚显微观测表明,淀粉粒在叶绿体中逐渐积累,类囊体片层结构被挤散,叶绿体膜被撑破。生理与生化分析表明衰老过程伴随着光合作用速率下降,光合色素降解加速,呼吸代谢的增加,这可能与衰老叶片中叶绿体逐渐崩塌和细胞膜透性增加相一致。iTRAQ标记方法共检测到不同发育阶段432个差异表达蛋白质,其中注释到308个与多种生命过程相关。蛋白差异富集分析表明,烟草叶片衰老过程中与光合作用等合成代谢相关蛋白多下调表达,而逆境反应及呼吸作用等分解代谢相关蛋白多上调表达。     

松油烯-4-醇光学异构体及外消旋体对家蝇的杀虫活性比较

【目的】比较松油烯-4-醇光学异构体对家蝇 Musca domestica 的熏蒸活性差异,为其光学异构体的应用提供理论依据。【方法】以家蝇4日龄成虫为供试昆虫,采用三角瓶熏蒸法比较测定了松油烯-4-醇光学异构体和外消旋体对其的熏蒸与击倒活性,并测定了松油烯-4-醇光学异构体及外消旋体对家蝇头部Na⁺ , K⁺-ATPase活性的影响。【结果】松油烯-4-醇外消旋体对家蝇的熏蒸活性和击倒活性最强,右旋异构体次之,左旋异构体最差,外消旋体、右旋异构体和左旋异构体对家蝇的致死中浓度（LC₅₀）分别为2.5,2.9和3.7 μL/L;在LC₉₀ 剂量下的击倒中时（KT₅₀）分别为12.6,16.7和18.9 min;松油烯-4-醇光学异构体及外消旋体均可显著抑制Na⁺, K⁺-ATPase的活性,活体条件下,松油烯-4-醇光学异构体及外消旋体对Na⁺, K⁺-ATPase活性的抑制作用随着中毒症状的加剧而增强,具有时间效应,其中左旋异构体的抑制作用最强;离体条件下,松油烯-4-醇光学异构体及外消旋体对Na⁺, K⁺-ATPase活性的抑制作用具有浓度依赖效应,其中外消旋体对Na⁺, K⁺-ATPase活性的抑制能力最强,明显高于同浓度下的右旋异构体和左旋异构体。【结论】松油烯-4-醇的光学异构体对家蝇的杀虫活性存在差异,外消旋体的活性明显高于异构体单体。开发松油烯-4-醇类杀虫剂,应以光学异构体的混合物作为有效成分。     

Productive and non-productive complexes in cytochrome P450-containing systems

The equilibrium dissociation constants K_D, the complex association / dissociation rate constants (k _on /k _off) and lifetimes of the complexes of redox partners were measured for three cytochrome P450-containing monooxygenase systems (P450cam, P450scc, and P450 2B4) under hydroxylation conditions. The Q parameter representing the ratio of protein-protein complex lifetime (τ _lT ) to time required for a single hydroxylation cycle (τ_turnover) was introduced for estimation of productivity of complexes formed within the systems studied. The Q parameter was insignificantly changed upon transition from the oxidation to hydroxylation conditions. Lifetimes (τ _lT ) for the binary complexes formed within the P450cam and the P450scc systems obligatory requiring an intermediate electron transfer protein between the reductase and cytochrome P450 could not realize hydroxylation reactions for substrates with known τ_turnover and so they were non-productive while the binary complexes formed within the P450 2B4 system, not requiring such intermediate electron-transfer protein, appeared to be productive. Formation of ternary complexes was demonstrated under hydroxylation conditions in all three systems. Analysis of Q values led to the conclusion that the ternary complexes formed within the P450cam and the P450scc systems were productive. In the case of the P450 2B4 system, more than half (about 60%) ternary complexes were also found to be productive.