BACKGROUND & AIMS: :In order to survive and adapt to the environment, it is imperative for fungi to be able to sense and respond to changes in extracellular pH conditions. In ascomycetes, sensing of extracellular pH is mediated by the Pal pathway resulting in activation of the PacC transcription factor at alkaline pH. The role of PacC in regulating fungal virulence and pathogenicity has been described in several pathogenic fungi but to date not in a symbiotic fungus. Epichloë festucae is a biotrophic fungal endophyte that forms a stable mutualistic interaction with Lolium perenne. In this study, pacC deletion (ΔpacC) and dominant active (pacC(C)) mutants were generated in order to study the cellular roles of PacC in E. festucae. Deletion of pacC resulted in increased sensitivity of the mutant to salt-stress but surprisingly did not affect the ability of the mutant to grow under alkaline pH conditions. Alkaline pH was observed to induce conidiation in wild-type E. festucae but not in the ΔpacC mutant. On the other hand the pacC(C) mutant had increased conidiation at neutral pH alone. Null pacC mutants had no effect on the symbiotic interaction with ryegrass plants whereas the pacC(C) mutant increased the tiller number. Examination of the growth of the pacC(C) mutant in the plant revealed the formation of aberrant convoluted hyphal structures and an increase in hyphal breakage, which are possible reasons for the altered host interaction phenotype.

背景与目标： : 为了生存并适应环境，真菌必须能够感知并响应细胞外pH条件的变化。在子囊菌中，细胞外pH的感测是由Pal途径介导的，从而导致碱性pH下PacC转录因子的激活。在几种致病真菌中已经描述了PacC在调节真菌毒力和致病性中的作用，但迄今为止在共生真菌中尚未描述。Epichlo ë festucae是一种生物营养真菌内生菌，与黑麦草形成稳定的相互作用。在这项研究中，产生了pacC缺失 (Δ pacC) 和显性活性 (PacC (C)) 突变体，以研究pacC在festucae中的细胞作用。pacC的缺失导致突变体对盐胁迫的敏感性增加，但令人惊讶的是，它不影响突变体在碱性pH条件下生长的能力。观察到碱性pH在野生型E. festucae中诱导分生孢子，但在 Δ pacc突变体中不诱导分生孢子。另一方面，仅在中性pH下，pacC(C) 突变体的分生孢子增加。无效的pacC突变体对与黑麦草植物的共生相互作用没有影响，而pacC(C) 突变体增加了分till数。对植物中pacC(C) 突变体的生长进行的检查表明，形成了异常的回旋菌丝结构，菌丝断裂增加，这可能是宿主相互作用表型改变的原因。

BACKGROUND & AIMS: :An Aspergillus niger strain has been constructed in which the pH-dependent regulatory gene, pacC, was disrupted. The pacC gene of A. niger, like that of A. nidulans, is involved in the regulation of acid phosphatase expression. Disruptants were identified by a reduction in acid phosphatase staining of colonies. Southern analysis demonstrated integration of the disruption plasmid at the pacC locus and Northern analysis showed that the disruption strain produced a truncated pacC mRNA of 2.2 kb (as compared to 2.8 kb in the wild type). The strain carrying the pacC disruption was used to assign the pacC gene to linkage group IV; this was confirmed by CHEF electrophoresis and Southern analysis. This strain further allowed us to determine which extracellular enzyme and transport systems are under the control of pacC in A. niger. Expression of the A. niger pacC wild-type gene and the truncated pacC gene showed that, in contrast to the auto-regulated wild-type expression, which was elevated only at alkaline pH, the truncated pacC gene was deregulated, as high-level expression occurred regardless of the pH of the culture medium. Analysis of the phosphatase spectrum by isoelectric focussing and enzyme activity staining both in the wild-type and the pacC disruptant showed that at least three acid phosphatases are regulated by the pacC. For the single alkaline phosphatase no pH regulation was observed.

背景与目标： : 已经构建了黑曲霉菌株，其中pH依赖性调节基因pacC被破坏。与A.Niddulans一样，黑曲霉的pacC基因参与酸性磷酸酶表达的调节。通过减少菌落的酸性磷酸酶染色来鉴定破坏物。Southern分析证明了破坏质粒在pacC基因座上的整合，Northern分析表明，破坏菌株产生了2.2 kb的截短的pacC mRNA (与野生型中的2.8 kb相比)。携带pacC破坏的菌株用于将pacC基因分配给IV连锁组; 这已通过CHEF电泳和Southern分析得到证实。该菌株进一步使我们能够确定黑曲霉中哪些胞外酶和转运系统处于pacC的控制之下。A. niger pacC野生型基因和截短的pacC基因的表达表明，与仅在碱性pH下升高的自动调节野生型表达相反，截短的pacC基因被解除管制，无论培养基的ph值如何，都会发生高水平的表达。通过等电聚焦和酶活性染色对野生型和pacC破坏物中的磷酸酶谱进行分析表明，至少三种酸性磷酸酶受pacC调节。对于单一碱性磷酸酶，未观察到pH调节。

BACKGROUND & AIMS: INTRODUCTION:Intrathecal therapy is an important part of the pain treatment algorithm for chronic disease states. The use of this option is a viable treatment strategy, but it is inherent for pain physicians to understand risk assessment and mitigation. In this manuscript, we explore evidence and mitigating strategies to improve safety with intrathecal therapy. METHODS:A robust literature search was performed covering January 2011 to October 9, 2016, in PubMed, Embase, MEDLINE, Biomed Central, Google Scholar, Current Contents Connect, and International Pharmaceutical Abstracts. The information was cross-referenced and compiled for evidence, analysis, and consensus review, with the intent to offer weighted recommendations and consensus statements on safety for targeted intrathecal therapy delivery. RESULTS:The Polyanalgesic Consensus Conference has made several best practice recommendations to improve care and reduce morbidity and mortality associated with intrathecal therapy through all phases of management. The United States Prevention Service Task Force evidence level and consensus strength assessments are offered for each recommendation. CONCLUSION:Intrathecal therapy is a viable and relatively safe option for the treatment of cancer- and noncancer-related pain. Continued research and expert opinion are required to improve our current pharmacokinetic and pharmacodynamic model of intrathecal drug delivery, as this will undoubtedly improve safety and efficacy.

背景与目标：

BACKGROUND & AIMS: :Destruction of the pulmonary epithelium is a major feature of lung diseases caused by the mould pathogen Aspergillus fumigatus. Although it is widely postulated that tissue invasion is governed by fungal proteases, A. fumigatus mutants lacking individual or multiple enzymes remain fully invasive, suggesting a concomitant requirement for other pathogenic activities during host invasion. In this study we discovered, and exploited, a novel, tissue non-invasive, phenotype in A. fumigatus mutants lacking the pH-responsive transcription factor PacC. Our study revealed a novel mode of epithelial entry, occurring in a cell wall-dependent manner prior to protease production, and via the Dectin-1 β-glucan receptor. ΔpacC mutants are defective in both contact-mediated epithelial entry and protease expression, and significantly attenuated for pathogenicity in leukopenic mice. We combined murine infection modelling, in vivo transcriptomics, and in vitro infections of human alveolar epithelia, to delineate two major, and sequentially acting, PacC-dependent processes impacting epithelial integrity in vitro and tissue invasion in the whole animal. We demonstrate that A. fumigatus spores and germlings are internalised by epithelial cells in a contact-, actin-, cell wall- and Dectin-1 dependent manner and ΔpacC mutants, which aberrantly remodel the cell wall during germinative growth, are unable to gain entry into epithelial cells, both in vitro and in vivo. We further show that PacC acts as a global transcriptional regulator of secreted molecules during growth in the leukopenic mammalian lung, and profile the full cohort of secreted gene products expressed during invasive infection. Our study reveals a combinatorial mode of tissue entry dependent upon sequential, and mechanistically distinct, perturbations of the pulmonary epithelium and demonstrates, for the first time a protective role for Dectin-1 blockade in epithelial defences. Infecting ΔpacC mutants are hypersensitive to cell wall-active antifungal agents highlighting the value of PacC signalling as a target for antifungal therapy.

背景与目标： : 肺上皮的破坏是由霉菌病原体烟曲霉引起的肺部疾病的主要特征。尽管人们普遍认为组织入侵是由真菌蛋白酶控制的，但缺乏单个或多种酶的烟曲霉突变体仍然具有完全的侵入性，这表明在宿主入侵过程中还需要其他致病活动。在这项研究中，我们在缺乏pH响应转录因子PacC的烟曲霉突变体中发现并开发了一种新颖的组织非侵入性表型。我们的研究揭示了一种新的上皮进入模式，在蛋白酶产生之前通过Dectin-1的 β-葡聚糖受体以细胞壁依赖性方式发生。Δ pacc突变体在接触介导的上皮进入和蛋白酶表达中均有缺陷，并且在白细胞减少症小鼠中的致病性显着减弱。我们结合了鼠感染模型，体内转录组学和人肺泡上皮的体外感染，以描述两个主要且依次作用的依赖于PacC的过程，这些过程影响整个动物的上皮完整性和组织侵袭。我们证明烟曲霉孢子和发芽被上皮细胞以接触，肌动蛋白，细胞壁和Dectin-1依赖性方式内在化，并且 Δ pacc突变体在发芽生长过程中异常重塑细胞壁，无法进入上皮细胞，无论是在体外还是体内。我们进一步表明，PacC在白细胞减少的哺乳动物肺的生长过程中充当分泌分子的整体转录调节剂，并描述了侵袭性感染期间表达的分泌基因产物的完整队列。我们的研究揭示了一种组合的组织进入模式，该模式取决于肺上皮的顺序和机械上不同的扰动，并首次证明了Dectin-1阻断在上皮防御中的保护作用。感染 Δ PacC突变体对细胞壁活性抗真菌剂过敏，突出了PacC信号传导作为抗真菌治疗靶标的价值。

BACKGROUND & AIMS: :The Aspergillus PacC transcription factor undergoes proteolytic activation in response to alkaline ambient pH. In acidic environments, the 674 residue translation product adopts a 'closed' conformation, protected from activation through intramolecular interactions involving the < or = 150 residue C-terminal domain. pH signalling converts PacC to an accessible conformation enabling processing cleavage within residues 252--254. We demonstrate that activation of PacC requires two sequential proteolytic steps. First, the 'closed' translation product is converted to an accessible, committed intermediate by proteolytic elimination of the C-terminus. This ambient pH-regulated cleavage is required for the final, pH-independent processing reaction and is mediated by a distinct signalling protease (possibly PalB). The signalling protease cleaves PacC between residues 493 and 500, within a conserved 24 residue 'signalling protease box'. Precise deletion or Leu498Ser substitution prevents formation of the committed and processed forms, demonstrating that signalling cleavage is essential for final processing. In contrast, signalling cleavage is not required for processing of the Leu340Ser protein, which lacks interactions preventing processing. In its two-step mechanism, PacC processing can be compared with regulated intramembrane proteolysis.

背景与目标： : 曲霉PacC转录因子在碱性环境pH下发生蛋白水解激活。在酸性环境中，674残基翻译产物采用 “封闭” 构象，通过涉及 <或 = 150残基C-末端结构域的分子内相互作用保护免于活化。pH信号传导将PacC转化为能够在残基252-254内处理裂解的可及构象。我们证明了PacC的激活需要两个连续的蛋白水解步骤。首先，通过蛋白水解消除C末端，将 “封闭” 的翻译产物转化为可访问的，承诺的中间体。这种环境pH调节的裂解是最终的，与pH无关的加工反应所必需的，并且由不同的信号蛋白酶 (可能是PalB) 介导。信号蛋白酶在保守的24个残基 “信号蛋白酶盒” 内切割残基493和500之间的PacC。精确的删除或Leu498Ser替换可防止提交和处理的形式的形成，这表明信号裂解对于最终加工至关重要。相反，Leu340Ser蛋白的加工不需要信号裂解，因为它缺乏阻止加工的相互作用。在其两步机制中，可以将PacC处理与受调控的膜内蛋白水解进行比较。

BACKGROUND & AIMS: :A homologue of the gene encoding the transcription factor Rim101 (PacC), involved in pH signal transduction in fungi, was identified in the pathogenic basidiomycete Ustilago maydis. The gene (RIM101) encodes a protein of 827 amino acid residues, which shows highest similarity to PacC proteins from Fusarium oxysporum and Aspergillus niger. The gene had the capacity to restore protease activity to rim101 mutants from Yarrowia lipolytica, confirming its homologous function, and was expressed at both acid and neutral pH. Null Deltarim101 mutants were not affected in the in vitro pH-induced dimorphic transition, their growth rate, resistance to hypertonic sorbitol or KCl stress, and pathogenicity. However, similar to pacC (rim101) mutants in other fungi, they displayed a pleiotropic phenotype with alterations in morphogenesis, impairment in protease secretion, and increased sensitivity to Na+ and Li+ ions. Other phenotypic characteristics not previously reported in fungal pacC (rim101) mutants (morphological changes, increased sensitivity to lytic enzymes, and augmented polysaccharide secretion) were also observed in U. maydis mutants. All these modifications were alleviated by transformation with the wild-type gene, confirming that all were the result of mutation in RIM101. These data indicate that the Pal/Rim pathway is functional in U. maydis (and probably in other basidiomycetes) and plays complex roles in pH-sensing phenomena, as occurs in ascomycetes and deuteromycetes.

背景与目标： : 在致病性担子菌Ustilago maydis中鉴定了编码转录因子Rim101 (PacC) 的基因的同源物，该基因参与真菌中的pH信号转导。该基因 (RIM101) 编码827个氨基酸残基的蛋白质，该蛋白质与尖孢镰刀菌和黑曲霉的PacC蛋白具有最高的相似性。该基因具有恢复解脂耶氏酵母rim101突变体的蛋白酶活性的能力，证实了其同源功能，并在酸性和中性pH下表达。无效的Deltarim101突变体在体外pH诱导的二态转变，其生长速率，对高渗山梨醇或KCl胁迫的抗性以及致病性均不受影响。然而，与其他真菌中的pacC (rim101) 突变体相似，它们显示出多效性表型，其形态发生改变，蛋白酶分泌受损以及对Na和Li离子的敏感性增加。在U. maydis突变体中还观察到以前未在真菌pacC (rim101) 突变体中报道的其他表型特征 (形态变化，对裂解酶的敏感性增加和多糖分泌增加)。所有这些修饰都通过与野生型基因的转化而得到缓解，证实所有这些修饰都是rim101突变的结果。这些数据表明Pal/Rim途径在U. maydis (可能在其他担子菌中) 中起作用，并且在pH感应现象中起着复杂的作用，例如在子囊菌和半知菌中。

BACKGROUND & AIMS: :Penicillium expansum, the causal agent of blue mould rot, causes severe post-harvest fruit maceration simultaneously with the secretion of d-gluconic acid (GLA) and the mycotoxin patulin in colonized tissue. The factor(s) inducing patulin biosynthesis during colonization of the host acidic environment is unclear. During the colonization of apple fruit in vivo and growth in culture, P. expansum secretes pH-modulating GLA and ammonia. Although patulin and its possible opportunistic precursor GLA accumulate together during fungal development, ammonia is detected on the colonized tissue's leading edge and after extended culture, close to patulin accumulation. Here, we demonstrate ammonia-induced transcript activation of the global pH modulator PacC and patulin accumulation in the presence of GLA by: (i) direct exogenous treatment of P. expansum growing on solid medium; (ii) direct exogenous treatment on colonized apple tissue; (iii) growth under self-ammonia production conditions with limited carbon; and (iv) analysis of the transcriptional response to ammonia of the patulin biosynthesis cluster. Ammonia induced patulin accumulation concurrently with the transcript activation of pacC and patulin biosynthesis cluster genes, indicating the regulatory effect of ammonia on pacC transcript expression under acidic conditions. Electrophoretic mobility shift assays using P. expansum PacC and antibodies to the different cleaved proteins showed that PacC is not protected against proteolytic signalling at pH 4.5 relative to pH 7.0, but NH4 addition did not further enhance its proteolytic cleavage. Ammonia enhanced the activation of palF transcript in the Pal pathway under acidic conditions. Ammonia accumulation in the host environment by the pathogen under acidic pH may be a regulatory cue for pacC activation, towards the accumulation of secondary metabolites, such as patulin.

背景与目标： : 青霉青霉是蓝霉腐烂的病原体，在定植组织中分泌d-葡萄糖酸 (GLA) 和霉菌毒素棒曲霉素的同时，引起了严重的收获后浸渍。在宿主酸性环境定植过程中诱导棒曲霉素生物合成的因素尚不清楚。在苹果果实体内定植和培养过程中，P. expansum分泌调节pH的GLA和氨。尽管棒曲霉素及其可能的机会性前体GLA在真菌发育过程中积聚在一起，但在定植组织的前缘和长时间培养后检测到氨，接近棒曲霉素的积累。在这里，我们通过以下方式证明了氨诱导的转录激活全球pH调节剂PacC和棒曲霉素在GLA存在下的积累 :( i) 在固体培养基上生长的P. expansum的直接外源处理; (ii) 在定植的苹果组织上的直接外源处理; (iii) 在碳有限的自氨生产条件下生长; (iv) 分析棒曲霉素生物合成簇对氨的转录反应。氨诱导棒曲霉素的积累与pacC和棒曲霉素生物合成簇基因的转录激活同时发生，表明氨在酸性条件下对pacC转录本表达的调节作用。使用P. Expansum  pacc和针对不同裂解蛋白的抗体的电泳迁移率位移测定表明，相对于pH 7.0，在pH 4.5时，PacC不能防止蛋白水解信号传导，但NH4的添加不能进一步增强其蛋白水解切割。在酸性条件下，氨增强了Pal途径中palF转录本的活化。在酸性pH下，病原体在宿主环境中积累的氨可能是pacC活化的调节线索，从而导致次级代谢产物 (例如棒曲霉素) 的积累。

BACKGROUND & AIMS: :Cellulosic biomass represents a valuable potential substitute for fossil-based fuels. As such, there is a strong need to develop efficient biotechnological processes for the enzymatic hydrolysis of cellulosic biomass via the optimization of cellulase production by fungi. Ambient pH is an important factor affecting the industrial production of cellulase. In the present study, we demonstrate that several Aspergillus nidulans genes encoding cellulolytic enzymes are regulated by Pal-PacC-mediated pH signaling, as evidenced by the decreased cellulase productivity of the palC mutant and pacC deletants of A. nidulans. The deletion of pacC was observed to result in delayed induction and decreased expression of the cellulase genes based on time course expression analysis. The genome-wide identification of PacC-regulated genes under cellobiose-induced conditions demonstrated that genes expressed in a PacC-dependent manner included 82 % of ClrB (a transcriptional activator of the cellulase genes)-regulated genes, including orthologs of various transporter and β-glucosidase genes considered to be involved in cellobiose uptake or production of stronger inducer molecules. Together with the significant overlap between ClrB- and PacC-regulated genes, the results suggest that PacC-mediated regulation of the cellulase genes involves not only direct regulation by binding to their promoter regions but also indirect regulation via modulation of the expression of genes involved in ClrB-dependent transcriptional activation. Our findings are expected to contribute to the development of more efficient industrial cellulase production methods.

背景与目标： : 纤维素生物质是化石燃料的有价值的潜在替代品。因此，迫切需要通过优化真菌产生的纤维素酶来开发有效的生物技术方法，以酶促水解纤维素生物质。环境ph值是影响纤维素酶工业生产的重要因素。在本研究中，我们证明了编码纤维素分解酶的几个构巢曲霉基因受Pal-PacC介导的pH信号传导的调节，这可以通过下降的纤维素酶生产率证明。根据时程表达分析，观察到pcc的缺失导致纤维素酶基因的诱导延迟和表达降低。在纤维二糖诱导的条件下对PacC调节基因的全基因组鉴定表明，以PacC依赖性方式表达的基因包括ClrB (纤维素酶基因的转录激活因子) 调节基因的82%，包括被认为参与纤维二糖摄取或产生更强诱导剂分子的各种转运蛋白和 β-葡萄糖苷酶基因的直系同源物。再加上ClrB和PacC调节的基因之间的显着重叠，结果表明，PacC介导的纤维素酶基因的调控不仅涉及通过结合其启动子区域而直接调控，而且还涉及通过调节涉及的基因表达的间接调控ClrB依赖性转录激活。我们的发现有望有助于开发更有效的工业纤维素酶生产方法。

BACKGROUND & AIMS: :The transcription factor PacC/Rim101 participates in environmental pH adaptation, development and secondary metabolism in many fungi, but whether PacC/Rim101 contributes to fungal adaptation to environmental stress remains unclear. In our previous study, a homologous gene of PacC/Rim101 was identified, and PacC-silenced strains of the agaricomycete Ganoderma lucidum were constructed. In this study, we further investigated the functions of PacC in G. lucidum and found that PacC-silenced strains were hypersensitive to environmental stresses, such as osmotic stress, oxidative stress and cell wall stress, compared with wild-type (WT) and empty-vector control (CK) strains. In addition, transmission electron microscopy images of the cell wall structure showed that the cell walls of the PacC-silenced strains were thinner (by approximately 25-30%) than those of the WT and CK strains. Further analysis of cell wall composition showed that the β-1,3-glucan content in the PacC-silenced strains was only approximately 78-80% of that in the WT strain, and the changes in β-1,3-glucan content were consistent with downregulation of glucan synthase gene expression. The ability of PacC to bind to the promoters of glucan synthase-encoding genes confirms that PacC transcriptionally regulates these genes.

背景与目标： 转录因子PacC/Rim101参与了许多真菌的环境pH适应，发育和次生代谢，但PacC/Rim101是否有助于真菌对环境胁迫的适应仍不清楚。在我们先前的研究中，鉴定了PacC/Rim101的同源基因，并构建了伞菌灵芝的PacC沉默菌株。在这项研究中，我们进一步研究了灵芝中PacC的功能，发现与野生型 (WT) 和空载体控制 (CK) 菌株相比，PacC沉默菌株对环境胁迫 (如渗透应激，氧化应激和细胞壁应激) 高度敏感。此外，细胞壁结构的透射电子显微镜图像显示，经PacC沉默的菌株的细胞壁比WT和CK菌株的细胞壁薄 (约25-30%)。细胞壁组成的进一步分析表明，PacC沉默菌株中的 β-1，3-葡聚糖含量仅为WT菌株的约78-80%，并且 β-1，3-葡聚糖含量的变化与葡聚糖合酶基因表达的下调一致。PacC与葡聚糖合酶编码基因启动子结合的能力证实了PacC在转录上调节这些基因。

BACKGROUND & AIMS: BACKGROUND:In fungi, environmental pH is an important signal for development, and successful host colonization depends on homeostasis. Surprisingly, little is known regarding the role of pH in fungal-fungal interactions. Species of Trichoderma grow as soil saprobes but many are primarily mycotrophic, using other fungi as hosts. Therefore, Trichoderma spp. are studied for their potential in biocontrol of plant diseases. Particularly in alkaline soil, pH is a critical limiting factor for these biofungicides, whose optimal growth pH is 4-6. Gaining an understanding of pH adaptability is an important step in broadening the activity spectrum of these economically important fungi. RESULTS:We studied the pH-responsive transcription factor PacC by gene knockout and by introduction of a constitutively active allele (pacCc). ΔpacC mutants exhibited reduced growth at alkaline pH, while pacCc strains grew poorly at acidic pH. In plate confrontation assays ΔpacC mutants showed decreased ability to compete with the plant pathogens Rhizoctonia solani and Sclerotium rolfsii. The pacCc strain exhibited an overgrowth of R. solani that was comparable to the wild type, but was unable to overgrow S. rolfsii. To identify genes whose expression is dependent on pH and pacC, we designed oligonucleotide microarrays from the transcript models of the T. virens genome, and compared the transcriptomes of wild type and mutant cultures exposed to high or low pH. Transcript levels from several functional classes were dependent on pacC, on pH, or on both. Furthermore, the expression of a set of pacC-dependent genes was increased in the constitutively-active pacCc strain, and was pH-independent in some, but not all cases. CONCLUSIONS:PacC is important for biocontrol-related antagonism of other fungi by T. virens. As much as 5% of the transcriptome is pH-dependent, and of these genes, some 25% depend on pacC. Secondary metabolite biosynthesis and ion transport are among the relevant gene classes. We suggest that ΔpacC mutants may have lost their full biocontrol potential due to their inability to adapt to alkaline pH, to perceive ambient pH, or both. The results raise the novel possibility of genetically manipulating Trichoderma in order to improve adaptability and biocontrol at alkaline pH.

背景与目标：

BACKGROUND & AIMS: :The ability to respond to ambient pH is critical to the growth and virulence of the fungal pathogen Candida albicans. This response entails the differential expression of several genes affecting morphogenesis. To investigate the mechanism of pH-dependent gene expression, the C. albicans homolog of pacC, designated PRR2 (for pH response regulator), was identified and cloned. pacC encodes a zinc finger-containing transcription factor that mediates pH-dependent gene expression in Aspergillus nidulans. Mutants lacking PRR2 can no longer induce the expression of alkaline-expressed genes or repress acid-expressed genes at alkaline pH. Although the mutation did not affect growth of the cells at acid or alkaline pH, the mutants exhibited medium-conditional defects in filamentation. PRR2 was itself expressed in a pH-conditional manner, and its induction at alkaline pH was controlled by PRR1. PRR1 is homologous to palF, a regulator of pacC. Thus, PRR2 expression is controlled by a pH-dependent feedback loop. The results demonstrate that the pH response pathway of Aspergillus is conserved and that this pathway has been adapted to control dimorphism in C. albicans.

背景与目标： : 对环境ph值的反应能力对真菌病原体白色念珠菌的生长和毒力至关重要。这种反应需要影响形态发生的几个基因的差异表达。为了研究pH依赖性基因表达的机制，鉴定并克隆了pacC的白色念珠菌同源物PRR2 (用于pH反应调节剂)。pacC编码一种含锌指的转录因子，该因子介导构巢曲霉中pH依赖性基因表达。缺乏PRR2的突变体在碱性pH下不能再诱导碱性表达基因的表达或抑制酸性表达基因。尽管突变在酸性或碱性pH下不会影响细胞的生长，但突变体在丝状化方面表现出中等条件的缺陷。PRR2本身以pH条件方式表达，其在碱性pH下的诱导受prr1控制。PRR1与pacC的调节剂palF同源。因此，PRR2表达由pH依赖性反馈回路控制。结果表明，曲霉的pH反应途径是保守的，并且该途径已适应控制白色念珠菌的二态性。

BACKGROUND & AIMS: :The beta-lactam antibiotic penicillin is produced as an end product by some filamentous fungi only. It is synthesized from the amino acid precursors L-alpha-aminoadipic acid, L-cysteine, and L-valine. Previous data suggested that certain amino acids play a role in the regulation of its biosynthesis. Therefore, in this study the effects of externally added amino acids on both Aspergillus (Emericella) nidulans penicillin production and expression of the bidirectionally oriented biosynthesis genes acvA (pcbAB) and ipnA (pcbC) were comprehensively investigated. Different effects caused by amino acids on the expression of penicillin biosynthesis genes and penicillin production were observed. Amino acids with a major negative effect on the expression of acvA-uidA and ipnA-lacZ gene fusions, i.e., histidine, valine, lysine, and methionine, led to a decreased ambient pH during cultivation of the fungus. An analysis of deletion clones lacking binding sites for the pH-dependent transcriptional factor PACC in the intergenic regions between acvA-uidA and ipnA-lacZ gene fusions and in a pacC5 mutant (PacC5-5) suggested that the negative effects of histidine and valine on acvA-uidA expression were due to reduced activation by PACC under acidic conditions. These data also implied that PACC regulates the expression of acvA, predominantly through PACC binding site ipnA3. The repressing effect caused by lysine and methionine on acvA expression, however, was even enhanced in one of the deletion clones and the pacC5 mutant strain, suggesting that regulators other than PACC are also involved.

背景与目标： : β-内酰胺抗生素青霉素仅由某些丝状真菌作为最终产品生产。它是由氨基酸前体L-α-氨基己二酸，L-半胱氨酸和L-缬氨酸合成的。先前的数据表明，某些氨基酸在其生物合成的调节中起作用。因此，在本研究中，全面研究了外部添加氨基酸对曲霉 (Emericella) niddulans青霉素生产和双向定向生物合成基因acvA (pcbAB) 和ipnA (pcbC) 表达的影响。观察到氨基酸对青霉素生物合成基因表达和青霉素产生的不同影响。对acvA-uidA和ipnA-lacZ基因融合体的表达具有主要负面影响的氨基酸，即组氨酸，缬氨酸，赖氨酸和蛋氨酸，导致真菌培养过程中环境ph值降低。对acvA-uidA和ipnA-lacZ基因融合之间的基因间区域以及pacC5突变体 (PacC5-5) 中缺乏pH依赖性转录因子PACC结合位点的缺失克隆的分析表明，组氨酸和缬氨酸对acvA-uidA表达的负面影响是由于酸性条件。这些数据还暗示，PACC主要通过PACC结合位点ipna3调节acvA的表达。然而，赖氨酸和蛋氨酸对acvA表达引起的抑制作用甚至在一个缺失克隆和pcc5突变菌株中得到增强，这表明除PACC以外的其他调节剂也参与其中。

BACKGROUND & AIMS: BACKGROUND:Screening for coronary artery calcium with electron beam computed tomography (EBCT) has potential diagnostic and prognostic implications. Most prior research on this technology has been done on selected, high-risk populations. The goal of the Prospective Army Coronary Calcium (PACC) study is to determine the utility of EBCT for the detection of coronary calcium as a screening test for coronary artery disease and as an intervention for risk factor modification among young, asymptomatic, active-duty personnel undergoing the United States Army's Cardiovascular Screening Program. METHODS AND RESULTS:Three study designs will be used to address the objectives of this investigation: (1) a cross-sectional study of 2000 unselected, consecutive participants to determine the prevalence and extent of coronary calcification in the 40- to 45-year-old Army population, (2) a randomized, controlled trial with a 2 x 2 factorial design involving 1000 participants to assess the impact of EBCT information on several dimensions of patient behavior, with and without intensive risk factor case management, and (3) a prospective cohort study of 2000 participants followed for at least 5 years to establish the relation between coronary calcification and cardiovascular events in an unselected, "low-risk" (by conventional standards) Army population. CONCLUSIONS:We present a review of the literature on the clinical utility of EBCT, with a focus on the limited research in young, asymptomatic populations. The details of the PACC study (begun in October1998) are presented. The results of the PACC study will determine the clinical utility of EBCT in young, asymptomatic patients.

背景与目标：

BACKGROUND & AIMS: INTRODUCTION:Pain treatment is best performed when a patient-centric, safety-based philosophy is used to determine an algorithmic process to guide care. Since 2007, the International Neuromodulation Society has organized a group of experts to evaluate evidence and create a Polyanalgesic Consensus Conference (PACC) to guide practice. METHODS:The current PACC update was designed to address the deficiencies and innovations emerging since the previous PACC publication of 2012. An extensive literature search identified publications between January 15, 2007 and November 22, 2015 and authors contributed additional relevant sources. After reviewing the literature, the panel convened to determine evidence levels and degrees of recommendations for intrathecal therapy. This meeting served as the basis for consensus development, which was ranked as strong, moderate or weak. Algorithms were developed for intrathecal medication choices to treat nociceptive and neuropathic pain for patients with cancer, terminal illness, and noncancer pain, with either localized or diffuse pain. RESULTS:The PACC has developed an algorithmic process for several aspects of intrathecal drug delivery to promote safe and efficacious evidence-based care. Consensus opinion, based on expertise, was used to fill gaps in evidence. Thirty-one consensus points emerged from the panel considerations. CONCLUSION:New algorithms and guidance have been established to improve care with the use of intrathecal drug delivery.

背景与目标：

BACKGROUND & AIMS: BACKGROUND AND AIMS:Using a genetic predisposition score (GPS), integrating the additive associations of a set of single nucleotide polymorphisms (SNPs) with CHD, we examined the consequences of the joint presence of a high GPS and conventional risk factors (CRFs). METHODS AND RESULTS:We studied 11 SNPs at eight loci in 197 participants with prior CHD and 524 CHD-free subjects from the Boston Puerto Rican Health Study. Each polymorphism contributed 1 unit (high-risk allele homozygous), 0.5 units (heterozygous) and 0 units (low-risk allele homozygous) to the GPS. Odds ratio (OR) of CHD for those at high risk because of GPS (>5) and simultaneous presence of CRFs were estimated, compared with subjects at low risk, for both measurements. The mean score was higher in participants with prior CHD than those CHD-free (P=0.015), and the OR for CHD with a GPS>5 was 2.90 (P<0.001).The joint presence of a high GPS and each CRF was associated with higher risk of CHD. Compared to participants with high GPS, those with low GPS (