响应面法优化光合细菌Rhodospirillum Rubrum S1培养条件

文章首先用单因素法分别对光合细菌Rhodospirillum Rubrum S1的培养条件——温度、光照强度、接种量进行了考察,确定了最佳培养条件:温度30℃,光照强度3000lx,接种量15%。在此基础上,采用中心组合试验设计结合响应面分析法(Response Surface Methodology,RSM)对影响光合细菌Rhodospirillum Rubrum S1的发酵条件进行进一步优化。结果表明,主要影响因素的最佳条件:培养温度32.54℃,光照强度2995.85lx,接种量15.80%,光合细菌Rhodospirillum Rubrum S1的类胡萝卜素产量为16.72mg/L,与理论值16.86mg/L相差0.83%。因此,利用响应面分析法对光合细菌Rhodospirillum Rubrum S1的培养条件进行优化合理可行。

红糖槭(Acer rubrum L.)茎段离体培养研究

红糖槭(Acer rubrum L.)原产北美洲,近年引入我国。关于它的离体培养研究尚未见报道。我们采用自配的基本培养基与不同种类的植物激素及其不同配比组成试验组合对红糖械茎段进行离体培养试验,获得完整植株并移栽成活。

In vitro fungistatic activity of 36 traditional oriental medicines and their synergistic effect against Trichophyton rubrume

Objective: To investigate the fungistatic activity and synergistic effects of natural products and their constituents, including traditional oriental medicines(TOMs).Methods: Fungistatic activities of TOMs prepared by hot-water(115 ℃) or ethanol(70%; 40 ℃) extraction were determined by their minimum inhibitory concentration.To assess possible synergistic effects, minimum inhibitory concentrations of various combinations were evaluated.Results: By evaluating antifungal susceptibility of Trichophyton rubrum, which is a major causative fungus for several types of dermatophytosis, we confirmed that ethanol extracts were more active than hot-water extracts in 25 of the 36 TOMs, suggesting that the constituents with high hydrophobicity tend to contribute significantly to fungistatic activity.We selected four TOMs with high fungistatic activity, including Aucklandiae radix, Gentianae macrophyllae radix, Scutellariae radix, and Galla rhois, and their synergistic effects were investigated through the combination studies between TOMs or TOM-conventional drug terbinafine.In combinations between four TOMs, partial synergistic effects were observed in Aucklandiae radix–Galla rhois and Gentianae macrophyllae radix–Galla rhois combinations, as supported by the lowest fractional inhibitory concentration index value of 0.66 for both combinations.Furthermore, Galla rhois showed the strongest synergistic effect on growth inhibition of Trichophyton rubrumwith a fractional inhibitory concentration index value of 0.50 in combination with terbinafine.Conclusions: Our findings indicate that the combination of TOMs and TOM-terbinafine may be effective on treatment for chronic and recurrent dermatophytosis by improving fungistatic activity and led to decrease systemic toxicity in clinical practice.

Evapotranspiration and Above Ground Biomass of Acer rubrum from Liners to 8 m Tall Trees

To meet minimum spring flows, water management districts in Florida sought to make both agriculture and urban landscapes water efficient, which includes tree farms. Acer rubrum L. (red maple) trees are endemic to Central Florida and native to the eastern portion of the United States. Urban and suburban expansion has increased use of A. rubrum in landscape plantings and their production in nurseries. In Florida A. rubrum is planted around stormwater retention areas, but also in urban landscapes. To provide a basis for irrigation allocations both during production and in landscapes, daily actual evapotranspiration (ETA) for three red maple trees were measured with weighing lysimeters, beginning with rooted cuttings and continuing until trees averaged 8 m in height. Empirical models were derived to calculate ETA based on crown horizontal projected area or trunk caliper, adjusted daily by changes in reference evapotranspiration (ETo). Water use efficiency, based on carbon sequestered in above ground wood mass, was calculated at the end of five growing seasons. Average ETA to produce these maples was 29,107 L over 4.75 years, with an average water use efficiency of 1 kg dry mass of wood per 709 L of water lost by transpiration.

Human urine metabolome in response to Fermentum rubrum(Hongqu Capsules)by HILIC-UHPLC-QTOF

OBJECTIVE ″-omics″study represents an unbiased perspective to examine the bio-system to discover the novel biomarker(s)which might be overlooked when targeted analysis is performed instead.Urine,due to its ease of collection,minimal invasion involved and rich information of the downstream metabolites,has been extensively studied.Fermentum rubrum(Hongqu,HQ capsules)is a well-known traditional medicine with the claimed slimming effect which could be related to the reduction in the deposition of total cholesterol and glycerides.Lack in sufficient clinical evidence is always one of the leading reasons that hamper traditional medicines from gaining world-wide recognition.In our pursuit of scientific support for HQ capsules in managing obesity,we aim to examine the alteration of urinary metabolites in this small-scale human clinical research.METHODS 6 Chinese subjects were included and exposed to short-term administration of HQ capsules for 3d,twice daily,two capsules each dosing.The urine samples were collected for three days prior to the dosing and on the day following the last dosing.In total,96 urine samples were collected and then separated on hydrophilic interaction chromatography(HILIC)and analyzed by Agilent 6550 ESI-Quadrupoletime-of-flight(QTOF)mass spectrometer.RESULTS Under positive mode,two compounds were found to be present only in post-dosing urine,and one compound was significantly lowered in post-dosing samples.Those two compounds might be associated with the administered HQ capsules,which is formulated with multiherbal constituents.CONCLUSION Further elucidation on the structures of these compounds is needed to enable better understanding of the mechanism of HQ capsules in managing obesity.

Economic Value Evaluation of Fine Individual Plants of Ribes rubrum Linn. Based on AHP

In this experiment,the biological characteristics,the number of spikes per plant,the average number of panicles,the average spike length,and the average yield per plant of the wild Ribes rubrum Linn. were observed and determined and the analytic hierarchy process( AHP)was applied to evaluate the economic value of 8 different sources of excellent plants of R. rubrum Linn. The results indicated that " RXF02", " RXF03" and " RXM05" performed well in the number of spikes per plant,average number of panicles,average yield per plant,and cold resistance,and the comprehensive scores were 9. 68,9. 44 and 9. 41,respectively.

Majocchi’s granuloma caused by Trichophyton rubrum after facial injection with hyaluronic acid:A case report

BACKGROUND Facial cosmetic procedures become popular for people with a desire to have a younger appearance,and cosmetic technology has developed rapidly over the past several decades.However,increasing complications related to cosmetic injections have been reported,and infection is one of the most serious problems and can cause anxiety and facial injury.We here report a case of Majocchi’s granuloma(MG)caused by Trichophyton rubrum after facial injection of hyaluronic acid.CASE SUMMARY A 37-year-old woman presented to our hospital with a history of red papules,nodules,and abscesses on her left zygomatic arch for 2 mo.She had received a cosmetic injection of hyaluronic acid on the left side of her face prior to the appearance of the lesions.MG caused by Trichophyton rubrum after facial injection of hyaluronic acid was diagnosed based on morphology and molecular biological identification.In vitro antifungal susceptibility testing was conducted according to the Clinical and Laboratory Standards Institute M38-A2 method.Minimal inhibitory concentrations were used to evaluate the antifungal susceptibility.The antifungal agents and their minimal inhibitory concentrations for the strain were terbinafine(<0.5μg/mL),itraconazole(0.06μg/mL),amphotericin B(0.25μg/mL),fluconazole(32μg/mL),voriconazole(0.125μg/mL),posaconazole(0.125μg/mL),and isavuconazole(0.06μg/mL).We initially administered 250 mg/d oral terbinafine for 2 mo,but the patient still had painful papules,nodules and abscesses on her face.Then,we adjusted the treatment to itraconazole 400 mg/d for 8 wk based on the in vitro antifungal susceptibility testing results.The skin lesions improved significantly,and there was no recurrence during follow-up.CONCLUSION This case revealed that facial injection of hyaluronic acid may cause serious MG.Antifungal susceptibility testing should be considered in the treatment of MG caused by Trichophyton rubrum.

红花檵木Loropetalum chinense var.rubrum植物功能性状对光照强度的响应

通过研究不同光照条件下红花檵木功能性状的变化规律以及各功能性状间的相关关系,分析红花檵木植物功能性状对不同光照条件的响应及其对光照条件的适应性,以期为红花檵木在园林绿化工程中的科学配置提供理论依据。结果表明光照与红花檵木叶面积、比叶面积和茎比密度呈负相关,与叶干物质含量、叶片厚度、当年生枝条长度、枝干物质含量呈正相关,不同光照条件条件下比叶面积、叶厚度、叶干物质含量、当年生枝长等指标差异显著。光照条件对红花檵木各功能性状间的相关关系有影响,从全日照到半阴的光照条件,红花檵木功能性状间的相关关系增强。在半阴的光照条件下,具有大叶片的植株,叶片较薄,当年生枝条较长,干物质含量较低。

Systematic Identification of Acer rubrum bZIP Transcription Factors and Their Potential Role in Anthocyanin Accumulation under Low Temperature with Light

Acer rubrum is an important garden color-leafed plant.Its leaves will turn red in autumn,which is of great orna-mental value.The leaf color change in Acer rubrum is closely associated with anthocyanins accumulation.In anthocyanin synthesis and accumulation,various transcription factor families play significant regulatory roles,including the basic(region)leucine zipper(bZIP).However,there is no report on the systematic identification and functional analysis of the bZIPs in Acer rubrum.In this study,137 bZIPs distributed on 29 chromosomes of Acer rubrum were identified and renamed according to their locations on the chromosomes.According to the constructed bZIP phylogenetic tree of Arabidopsis thaliana and Acer rubrum,bZIPs were divided into 13 groups.Two pairs of bZIP genes were involved in tandem duplication,and 106 segmental duplication gene pairs were found.Cis-acting elements in the promoter region of these bZIP genes were analyzed.The results of promoter analysis showed that many elements were closely related to light conditions,hormone responses,and abiotic stress factors.Among them,the cis-acting elements related to light response were most abundant and prominent.The results of anthocyanin determination showed that anthocyanin contents in the leaves of Acer rubrum increased significantly under low temperature with light.In addition,gene expression analysis showed that compared to other ArbZIPs,ArbZIP137,ArbZIP136,ArbZIP114,ArbZIP130,and ArbZIP14 showed a more pronounced increase in gene expression both under low-temperature conditions and under light conditions.From the correlation analysis,there was a high correlation between ArbZIPs and several anthocyanin-regulated transcription factors,including ArMYBs,ArbHLH and ArWD40s.Conclusively,the bZIP genes in Acer rubrum were identified and analyzed,providing a foundational basis for future studies on their function and significantly enhancing our understanding of the color mechanism of Acer rubrum.

Synergistic activity of lawsone methyl ether in combination with some antibiotics and artocarpin against methicillin-resistant Staphylococcus aureus, Candida albicans, and Trychophyton rubrum

Objective: One appealing strategy to overcome and prevent resistant problem is the use of combined two or more antibacterial substances. Lawsone methyl ether(LME) is the naphthoquinone found in the leaves of Impatiens balsamina. The objective of this study is to determine the interaction of LME with some antibiotics(ampicillin, tetracycline, norfloxacin, and clotrimazole) and a natural compound, artocarpin against methicillin-resistant Staphylococcus aureus(MRSA), Candida albicans, and Trychophyton rubrum.Methods: A broth microdilution method was used to determine the minimum inhibition concentration(MIC). Synergistic effects were evaluated at their own MIC using the checkerboard method and time-kill assay.Results: LME showed moderate antibacterial activity against MRSA with MIC value of 15.6 μg/mL, and exhibited strong antifungal activities against T. rubrum and C. albicans with MIC values of 7.8 and 3.9 μg/m L,respectively. The interaction of LME with the natural compound artocarpin against MRSA produced a synergy with fractional inhibitory concentration index(FICI) value of 0.31, while the combination of LME and clotrimazole exhibited synergy against C. albicans and T. rubrum with FICI values of 0.38 and 0.24, respectively. The time-kill assays confirmed that the compounds in combination enhanced their antimicrobial activities against the resistant microorganisms with different degrees.Conclusion: LME in combination with clotrimazole exhibited synergy effect against C. albicans and T.rubrum. In combination with artocarpin, it showed synergy effect against MRSA.

A Trichophyton Rubrum Infection Model Based on the Reconstructed Human Epidermis - Episkin

Background： Trichophyton rubrum represents the most common infectious fungus responsible for dermatophytosis in human, but the mechanism involved is still not completely understood. An appropriate model constructed to simulate host infection is the prerequisite to study the pathogenesis of dernlatophytosis caused by T.. rubrum. In this study, we intended to develop a new T. rubrum infection model in vitro, using the three-dimensional reconstructed epidermis - EpiSkin, and to pave the way for further investigation of the mechanisms involved in T. rubrum infection. Methods： The reconstructed human epidermis （RHE） was infected by inoculating low-dose （400 conidia） and high-dose （4000 conidia） T. rubrum conidia to optimize the infection dose. During the various periods after infection, the samples were processed for pathological examination and scanning electron microscopy （SEM） observation. Results： The histological analysis of RHE revealed a fully differentiated epidermis with a functional stratum corneum, which was analogous to the normal human epidermis. The results of hematoxylin and eosin staining and the periodic acid-Schiff staining showed that the infection dose of 400 conidia was in accord with the pathological characteristics of host dermatophytosis caused by T. rubrum. SEM observations further exhibited the process of 77 ruhrum infection in an intuitionistic way, Conclusions： We established the T. rubrum infection model on RHE in vitro successfully. It is a promising model fbr further investigation of the mechanisms involved in T. rubrum infection.

Analysis of part of the Trichophyton rubrum ESTs

Trichophyton rubrum (T. rubrum) is the most common of the superficial fungi. In an effort to better understand the genetic and biochemical makeup of T. rubrum, we generated cDNA libraries from 3 growth stages and used these to isolate 4002 unique expressed sequence tags (ESTs). Sequence comparisons with the Genbank database allowed 1226 of the ESTs to be assigned putative functions or matched with homologs from other organisms. Of the remaining ESTs, 989 were only weakly similar to known sequences and 1787 had no identifiable functions, suggesting that they represent novel genes. We further analyzed the presence of several im-portant genes involved in the growth, metabolism, signal transduction, pathogenesis and drug resistance in T. rubrum. This information was used to newly elucidate important metabolic path-ways in T. rubrum. Taken together, our results should form the molecular basis for continued re-search on the physiological processes and pathogenic mechanisms of T. rubrum, and may lead to a better understanding of fungal drug resistance and identification of new drug targets.

Antifungal Activity of Aspidin BB from Dryopteris fragrans against Trichophyton rubrum Involved Inhibition of Ergosterol Biosynthesis

Objectives Aspidin BB, a typical phloroglucinol derivative from Dryopteris fragrans, possesses significant antifungal property. This study aimed to investigate potential mechanism of antifungal activity of Aspidin BB against Trichophyton rubrum which is the most common pathogens responsible for chronic dermatophytosis. Methods The minimum inhibitory concentration （MIC） ofAspidin BB against strains was determined by broth microdilution. The effects of Aspidin BB on ergosterol biosynthesis were investigated by content determination based on UPLC method. Besides, the effects of drugs on squalene epoxidase （SE） in T. rubrum cell membrane were analyzed. Results MIC value of Aspidin BB against T. rubrum was 25.0 IJg/mL. Aspidin BB reduced ergosterol content significantly, but no notable effect on squalene epoxidase activity. Conclusion The results suggested that Aspidin BB inhibited ergosterol biosynthesis. However, it was not squalene epoxidase but other components may sever as possible targets in ergosterol biosynthesis pathway.

Effects of laser irradiation on Trichophyton rubrum growth and ultrastructure

Background Trichophyton rubrum （T. rubrum） is the most common causative agent of dermatophytosis worldwide. In this study, we examined the effect of laser irradiation on the growth and morphology of T. rubrum. Methods Colonies of T. rubrum were isolated （one colony per plate）, and randomly assigned to 5 treatment groups： Q-switched 694 nm ruby laser treatment, long-pulsed Nd：YAG 1064 nm laser treatment, intense pulsed light （IPL） treatment, 308 nm excimer laser treatment and the blank control group without treatment. Standardized photographs were obtained from grown-up fungal plates prior to treatment. Colonies were then exposed to various wavelengths and fluences of laser light. To compare the growth of colonies, they were re-photographed under identical conditions three and six days post-treatment. To investigate the morphology of T. rubrum, scanning electron microscope （SEM） and transmission electron microscope （TEM） images were obtained from specimens exposed to 24 hours of laser treatment. Results Growth of T. rubrum colonies was significantly inhibited following irradiation by 694 nm Q-switched and 1064 nm long-pulsed Nd：YAG lasers. Other treatments exerted little or no effect. Q-switched laser irradiation exerted a stronger growth inhibitory effect than long-pulsed Nd：YAG laser irradiation. Following treatment by the Q-switched ruby laser system, T. rubrum hyphae became shrunken and deflated, and SEM images revealed rough, fractured hyphal surfaces, punctured with small destructive holes. TEM images showed that the hyphae were degenerating, as evidenced by the irregular shape of hyphae, rough and loose cell wall, and obscure cytoplasmic texture. Initially high electron density structure was visible in the cell; later, low-density structure appeared as a result of cytoplasmic dissolution. In contrast, the blank control group showed no obvious changes in morphology.

Regeneration and utilization of nutrients during collapse of a Mesodinium rubrum red tide and its influence on phytoplankton species composition

High-biomass red tides occur frequently in some semi-enclosed bays of Hong Kong where ambient nutrients are not high enough to support such a high phytoplankton biomass. These high-biomass red tides release massive inorganic nutrients into local waters during their collapse. We hypothesized that the massive inorganic nutrients released from the collapse of red tides would fuel growth of other phytoplankton species. This could influence phytoplankton species composition. We tested the hypothesis using a red tide event caused by Mesodinium rubrum(M. rubrum) in a semi-enclosed bay, Port Shelter. The red tide patch had a cell density as high as 5.0×10~5 cells L^(-1), and high chlorophyll a(63.71 μg L^(-1)). Ambient inorganic nutrients(nitrate:NO_3^-, ammonium: NH_4^+, phosphate: PO_4^(3-), silicate: SiO_4^(3-)) were low both in the red tide patch and the non-red-tide patch(clear waters outside the red tide patch). Nutrient addition experiments were conducted by adding all the inorganic nutrients to water samples from the two patches followed by incubation for 9 days. The results showed that the addition of inorganic nutrients did not sustain high M. rubrum cell density, which collapsed after day 1, and did not drive M. rubrum in the non-red-tide patch sample to the same high-cell density in the red tide patch sample. This confirmed that nutrients were not the driving factor for the formation of this red tide event, or for its collapse. The death of M. rubrum after day 1 released high concentrations of NO_3^-,PO_4^(3-), SiO_4^(3-), NH_4^+, and urea. Bacterial abundance and heterotrophic activity increased, reaching the highest on day 3 or 4, and decreased as cell density of M. rubrum declined. The released nutrients stimulated growth of diatoms, such as Chaetoceros affinis var. circinalis, Thalassiothrix frauenfeldii, and Nitzschia sp., particularly with additions of SiO_4^(3-) treatments, and other species. These results demonstrated that initiation of M. rubrum red tides in the bay was not directly driven by nutrients.However, the massive inorganic nutrients released from the collapse of the red tide could induce a second bloom in low-ambient nutrient water, influencing phytoplankton species composition.

Mechanisms underlying the vascular relaxation activities of Zingiber officinale var. rubrum in thoracic aorta of spontaneously hypertensive rats

Objective: To evaluate vasorelaxant and vasoconstriction effects of Zingiber officinale var. rubrum(ZOVR)on live rats and isolated aortic rings of spontaneously hypertensive rats(SHRs).Methods: Extracts of ZOVR were subjected to in-vivo antihypertensive screening using noninvasive blood pressures in SHRs. The most potent extract, ZOVR petroleum ether extract(ZOP) was then fractionated using n-hexane, chloroform and water. Isolated thoracic aortic rings were harvested and subjected to vascular relaxation studies of n-hexane fraction of ZOP(HFZOP) with incubation of different antagonists such as Nω-nitro-L-arginine methyl ester(L-NAME, 10 μmol/L), indomethacin(10 μmol/L), methylene blue(10 μmol/L), atropine(1 μmol/L), glibenclamide(10 μmol/L), prazosin(0.01 μmol/L), and propranolol(1 μmol/L).Results: During the screening of various ZOVR extracts, ZOP produced the most reduction in blood pressures of SHRs and so did HFZOP. HFZOP significantly decreased phenylephrine-induced contraction and enhanced acetylcholine-induced relaxation. L-NAME, indomethacin, methylene blue, atropine, and glibenclamide significantly potentiated the vasorelaxant effects of HFZOP. Propranolol and prazosin did not alter the vasorelaxant effects of HFZOP. HFZOP significantly suppressed the Ca2+-dependent contraction and influenced the ratio of the responses to phenylephrine in Ca2+-free medium.Conclusion: This study demonstrates that ZOP may exert an antihypertensive effect in the SHR model. Its possible vascular relaxation mechanisms involve nitric oxide and prostacyclin release, activation of cGMP-KATP channels, stimulation of muscarinic receptors, and transmembrane calcium channel or Ca2+release from intracellular stores. Possible active compounds that contribute to the vasorelaxant effects are 6-gingerol, 8-gingerol and 6-shogaol.

Global gene expression profiles for the growth phases of Trichophyton rubrum

Trichophyton rubrum （T. rubrum） is a common superficial fungus. Molecular and genetic studies of T. rubrum are still limited. In this paper, we report the global analysis of gene expression profiles at different growth phases using cDNA microarray technology. A total of 2044 differentially expressed genes were obtained and clustered into three expression patterns. Our data confirmed previous results that many mRNAs were pre-stored in the conidia of T. rubrum. Transcriptional profiling and function analysis showed that some glycolytic enzymes share similar expression patterns and may be coregulated during the transi- tion of growth phases. Some genes involved in small GTPase signaling pathways, and in cAMP-dependent and MAPK regulation pathways were induced in response to the growth dynamics of T. rubrum. Although the detailed biological roles of these Z rubrum genes are still unknown, our results suggest that these genes may be involved in regulation mechanisms in the life cycle of the fungus.

Exposure to heat-inactivated Trichophyton rubrum resulting in a limited immune response of human keratinocytes

Background Trichophyton rubrum （T. rubrum） represents the most important agent of dermatophytosis in humans. T. rubrum infection causes slight inflammation, and tends to be chronic and recurrent. It is suggested that it may result from the failure of epithelial cells to recognize T. rubrum effectively and initiate effective immune responses. The C-type lectin receptors （CLR） and toll-like receptors （TLR） are the two major pattern recognition receptors （PRRs） that recognize fungal components. Therefore, the purpose of the study was to analyze the expression of those PRRs and the cytokines in HaCaT cells stimulated with heat-inactivated T. rubrum conidia and hyphae, respectively. Methods HaCaT cells were unstimulated or stimulated with heat-inactivated T. rubrum conidia and hyphae （l×106 and 1.5×105 colony-forming unit （CFU） in 2 ml medium, respectively） for 6, 12 and 24 hours. The mRNA expression of PRRs involved in recognizing fungal pathogen-associated molecular patterns （PAMPs） and signaling molecules were measured by quantitative reverse transcription polymerase chain reaction （RT-PCR）. Meanwhile, surface toll-like receptor （TLR） 2, TLR4 and Dectin-1 were analyzed by fluorescence-activated cell sorter （FACS） 24 hours after treatment. The cytokines were detected in cell culture supernatants of HaCaT cells in 12 and 24 hours after treatment. Results HaCaT cells constitutively expressed mRNA of membrane-bound TLR1,2, 4 and 6, Dectinl and DC-SIGN, but not Dectin-2 or Mincle. Heat-killed T. rubrum did not significantly upregulate gene transcriptions of the PRRs of HaCaT cells. Heat-inactivated T. rubrum conidia significantly reduced the surface expression of TLR2 and Dectin-1, and suppressed the secretions of interferon-inducible protein-10 （IP-10） and monocyte chemotactic protein-1 （MCP-1） of HaCaT cells, while heat-killed T. rubrum hyphae significantly induced the secretions of IP-10 and MCP-I. Conclusion The cell-wall antigens of T. rubrum fail to activate transcriptional expression of PRRs and induce a lower immune response of HaCaT cells by limited cytokines secretion.

Comparison of a glucose consumption based method with the CLSI M38-A method for testing antifungal susceptibility of Trichophyton rubrum and Trichophyton mentagrophytes

Background The prevalence of dermatophytoses and the development of new antifungal agents has focused interest on susceptibility tests of dermatophytes. The method used universally for susceptibility tests of dermatophytes was published as document （M38-A） in 2002 by the Clinical and Laboratory Standards Institute （CLSI）, dealing with the standardization of susceptibility tests in filamentous fungi, though not including dermatophytes especially. However, it is not a very practical method for the clinical laboratory in routine susceptibility testing. In this test, we developed a novel rapid susceptibility assay --glucose consumption method （GCM） for dermatophytes. Methods In this study, we investigated the antifungal susceptibilities of dermatophytes to itraconazole （ITC）, voriconazole （VOC）, econazole nitrate （ECN） and terbinafine （TBF） by glucose consumption method （GCM）, in comparison to the Clinical and Laboratory Standards Institute （CLSI） M38-A method. Twenty-eight dermatophyte isolates, including Trichophyton rubrum （T. rubrum） （n=-14） and Trichophyton mentagrophytes （T. mentagrophytes） （n=-14）, were tested. In the GCM, the minimum inhibitory concentrations （MICs） were determined spectrophotometrically at 490 nm after addition of enzyme substrate color mix. For the CLSI method, the MICs were determined visually. Results Comparison revealed best agreement for TBF against T. mentagrophytes and T. rubrum, since MIC range, MIC50, and MIC90 were identical from two methods. However, for ITC and VOC, GCM showed wider MIC ranges and higher MICs than CLSI methods in most isolates. For ECN against T. rubrum, high MICs were tested by GCM （0.125-16 pg/ml） but not M38-A method （0.5-1 IJg/ml）. The overall agreements for all isolates between the two methods within one dilution and two dilutions for ITC, VOC, ECN and TBF was 53.6% and 75.0%, 57.1% and 75.0%, 82.1% and 89.3%, and 85.7 and 85.7%, respectively. Conclusion Measurement of glucose uptake can predict the susceptibility of T. rubrum and T. mentagrophytes to ECN and TBF.

Comparison between gene expression of conidia and germinating phase in Trichophyton rubrum

Trichophyton rubrum is a dominating superficial dermatophyte, whose conidial germination is corre- lated to pathopoiesis and a highly important developmental process. To investigate the changes of physiology, biochemistry and cytology during the germination, we selected 3364 function identified ESTs from T. rubrum cDNA library to construct cDNA microarrays, and compared the gene expression levels of conidia and germinating phase. Data analysis indicated that 335 genes were up-regulated during the germination, which mainly encoded translated, modified proteins and structural proteins. The constituents of cell wall and cell membrane were synthetized abundantly, suggesting that they are the foundation of cell morphogenesis. The ingredients of the two-component signal transduction sys- tem were up-regulated, presuming that they were important for the conidial germination. Genes of various metabolic pathways were expressed prosperously, especially the genes that participated in glycolysis and oxidative phosphorylation were up-regulated on the whole, demonstrating that in the environment with sufficient oxygen and glucose, conidia obtained energy through aerobic respiration. This paper provides important clues which are helpful to understanding the changes in gene expres- sion, signal conduction and metabolism characteristics during T. rubrum conidial germination, and possess significant meaning to the study of other superficial dermatophytes.