Genome-Wide Identification of the AP2/ERF Gene Family Involved in Active Constituent Biosynthesis in.

Tanshinones and phenolic acids are the major bioactive constituents in the traditional medicinal crop ; however, transcription factors (TFs) are seldom investigated with regard to their regulation of the biosynthesis of these compounds. Here a complete overview of the APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor family in is provided, including phylogeny, gene structure, conserved motifs, and gene expression profiles of different organs (root, stem, leaf, flower) and root tissues (periderm, phloem, xylem). In total, 170 AP2/ERF genes were identified and divided into five relatively conserved subfamilies, including AP2 (25 genes), DREB (61 genes), ethylene responsive factor (ERF; 79 genes), RAV (4 genes), and Soloist (1 gene). According to the distribution of bioactive constituents and the expression patterns of AP2/ERF genes in different organs and root tissues, the genes related to the biosynthesis of bioactive constituents were selected. On the basis of quantitative real-time polymerase chain reaction (qRT-PCR) analysis, coexpression analysis, and the prediction of -regulatory elements in the promoters, we propose that two genes ( and ) regulate tanshinone biosynthesis and two genes ( and ) participate in controlling phenolic acid biosynthesis. The genes related to tanshinone biosynthesis belong to the ERF-B3 subgroup. In contrast, the genes predicted to regulate phenolic acid biosynthesis belong to the ERF-B1 and ERF-B4 subgroups. These results provide a foundation for future functional characterization of AP2/ERF genes to enhance the biosynthesis of the bioactive compounds of .

First Report of Sclerotinia sclerotiorum Causing Sclerotinia Blight on Peanut (Arachis hypogaea) in Northeastern China.

Peanut, Arachis hypogaea L., is an important source of vegetable oil and protein in China with a planting area of 5 million ha and annual production of 16.2 million metric tons. In September of 2012, typical symptoms of Sclerotinia blight were first observed on peanut in Shuangcheng, Heilongjiang Province, China. Approximately 50% of the plants in a field were infected. Symptoms began as a chlorotic wilt on the foliage and developed into necrosis of basal stems and developed very quickly in the field. In advanced stages of the disease, stems and branches became bleached and eventually died. White, fluffy mycelium and black irregular sclerotia (3.5 to 5.4 mm diameter) were observed on the infected stems. Infected branches and pegs were shredded, and most pods dropped on the soil during harvest. To isolate the causal agent of the disease, sclerotia were collected from the field. Twenty sclerotia were surface disinfected in 1% NaOCl for 3 min, rinsed three times with sterile water, placed on potato sucrose agar (PSA) with 100 µg/ml streptomycin, and then incubated at 22°C in the dark for 10 days. Fungal DNA was extracted from mycelia with a TIANGEN DNAsecure Plant Kit (Beijing) and amplified by PCR with the universal fungal primer ITS1 and ITS4. PCR products of five replicates were sequenced and subjected to an NCBI BLAST search. The BLAST search revealed that our sequences (GenBank Accession No. KC935388) had 100% identity with reported sequences of Sclerotinia sclerotiorum. The isolates were identified as S. sclerotiorum (Lib.) de Bary based on mycelia, sclerotia, and rDNA sequence analysis. To conduct pathogenicity tests, nine potted peanut plants (1 month old) were each inoculated with a 5-mm-diameter disk of colonized PSA by placing the inoculum on the base of the stem. Plants were inoculated with a plug of non-colonized PSA as controls. All inoculated plants were covered with a plastic bag for 5 days to maintain high humidity and incubated at 22 to 24°C in the growth chamber with a 12-h photoperiod. After 2 days, the inoculated plants showed water-soaked brown symptoms on the stem base and whole plants wilted after 5 days under high moisture conditions. The symptoms were identical to those observed on peanut plants in Shuangcheng, Heilongjiang Province, whereas the control plants remained symptom-free. Re-isolation of the fungus from the inoculated plants confirmed that the causal agent was S. sclerotiorum. S. sclerotiorum has been reported on peanut in the United States and Argentina (1,2,3). To our knowledge, this is the first report of S. sclerotiorum as a pathogen causing Sclerotinia blight on peanut in northeastern China. References: (1) A. Marinelli et al. Int. J. Pest Manage. 44:251, 1998. (2) J. E. Woodward et al. Plant Dis. 90:111, 2006. (3) J. E. Woodward et al. Plant Dis. 92:1468, 2008.

Ethylene inhibited aflatoxin biosynthesis is due to oxidative stress alleviation and related to glutathione redox state changes in Aspergillus flavus.

The effect of 2-chloroethyl phosphoric acid (CEPA) on aflatoxin biosynthesis, the expression of aflatoxin biosynthetic genes, reactive oxygen species (ROS) formation, cellular redox status, and enzymes involved in glutathione consumption and regeneration in Aspergillus flavus was investigated. The results demonstrated that CEPA dose dependently inhibited aflatoxin B(1) production. The expression of two typical genes involved in aflatoxin biosynthesis, aflR and aflD, was reduced after CEPA treatment at 7 d. Meanwhile, CEPA significantly reduced ROS production and thiobarbituric acid reactive substances (TBARS), increased the ratio of reduced glutathione (GSH) and oxidized glutathione (GSSG) at 5, 6 and 7 d. The activities of glutathione peroxidase (GPx), glutathione reductase (GR), as well as glucose-6-phosphate dehydrogenase (G6PDH) were significantly inhibited after CEPA treatment at 5, 6 and 7 d. The present study suggested that ethylene reduced aflatoxin production is due to oxidative stress alleviation of fungal cells and is related to glutathione redox state changes.

A proposed referral centre based on HL7/XML.

With the growth of the Inteernet, hospitals have also applied HL7 (Health Level Seven) to exchange data between them. The referral system is identified as an appropriate application system. The effect of referral is to transfer the patient to a suitable hospital in a timely fashion, and to arrange appropriate treatment for the patient. Taking advantage of the Internet to exchange referral data can, not only accelerate the process of patient referral, but also avoid the unnecessary repeat examinations to decrease the waste of medical resources. This article builds up a referral-related message according to the HL7 standard, and develops a referral centre using the Internet environment, making use of XML (eXtensible Markup Language) standard to transform the referral-related data to XML format and exchange referral data between platforms. This electronic referral mechanism is expected to offer other hospitals experience of improved referral practice.

Security architecture for HL/7 message interchange.

The promotion of quality medical treatment is very important to the healthcare providers as well as to patients. It requires that the medical resources of different hospitals be combined to ensure that medical information is shared and that resources are not wasted. A computer-based patient record is one of the best methods to accomplish the interchange of the patient's clinical data. In our system, the Health Level/Seven (HL/7) format is used for the interchange of the clinical data, as it has been supported by many healthcare providers and become a â standard'. The security of the interchange of clinical data is a serious issue for people using the Internet for data communication. Several international well-developed security algorithms, models and secure policies are adopted in the design of a security handler for an HL/7 architecture. The goal of our system is to combine our security system with the end-to-end communication systems constructed from the HL/7 format to establish a safe delivery channel. A suitable security interchange environment is implemented to address some shortcomings in clinical data interchange. located at the application layer of the ISO/OSI reference model. The medical message components, sub-components, and related types of message event are the primary goals of the HL/7 protocols. The patient management system, the doctor's system for recording his advice, examination and diagnosis as well as any financial management system are all covered by the HL/7 protocols. Healthcare providers and hospitals in Taiwan are very interested in developing the HL/7 protocols as a common standard for clinical data interchange.

A preliminary study of cocaine absorption from the nasal mucosa.

OBJECTIVE/HYPOTHESIS: To determine factors affecting the safe use of topical cocaine for anesthesia and vasoconstriction during rhinologic surgery. STUDY DESIGN: Prospective, randomized study of the kinetics of cocaine absorption through human nasal mucosa in 12 consecutive patients without nasal mucosal disease who were having septoplasty or septorhinoplasty. METHODS: With patients under general anesthesia, cocaine was applied topically to each nasal cavity by using cottonoid pledgets. Group I received 4 mL of a 4% solution (160 mg) of cocaine for 10 min, Group II received 4 mL of a 4% solution (160 mg) of cocaine for 20 min, and Group III received 4 mL of a 10% solution (400 mg) of cocaine for 20 min. Absorption rate was determined by measuring serum cocaine concentration at intervals of 5, 10, 15, and 20 min. Residual cocaine was extracted from the pledgets and was analyzed quantitatively by using gas chromatography and mass spectroscopy. RESULTS: Of total cocaine applied, 35% was absorbed systemically: 17% was absorbed within 5 min, 25% within 10 min, and 32% within 15 min. Of the cocaine absorbed, 47% was absorbed within the first 5 min, 70% within 10 min, and 90% within 15 min. Two patients (16.6%), both in Group III, had intraoperative hypertension; one of these patients also had transient ventricular tachycardia. CONCLUSIONS: Although a 4% solution of cocaine applied to the nasal mucosa on cottonoid pledgets for 20 min is safe, we observed an idiosyncratic absorption rate four times greater than expected; therefore, we advise against topical use of a 10% cocaine solution for anesthesia and vasoconstriction during rhinologic surgery.

Septal pleomorphic adenoma masquerading as squamous cell carcinoma.

A 43-year-old woman had left midfacial pain and a left intranasal mass. Pathologists from three different institutions agreed that the biopsy specimens from the mass represented squamous cell carcinoma. The final pathologic diagnosis made from the resected specimen was pleomorphic adenoma of the septum. This report describes a case which was initially misdiagnosed on the basis of misleading tissue biopsy results, but the excised tumor was finally diagnosed as pleomorphic adenoma. Otolaryngologists treating an intranasal mass must be aware of the difficulty of correctly diagnosing the initial tissue biopsy specimen in order to guard against unnecessary radical surgery.

Audiometric comparison of Lassa fever hearing loss and idiopathic sudden hearing loss: evidence for viral cause.

A recently published prospective study on acute sensorineural deafness in Lassa fever among a West African population showed the audiometric pattern of a known virally induced hearing loss. Using the audiometric data from the patients with Lassa fever in that study, we analyzed and classified the initial hearing loss and final recovery into three groups by pure-tone average values and then did the same for 222 patients with idiopathic sudden hearing loss (SHL) in our study. Statistical analyses of the severity of initial hearing loss and the hearing recovery pattern indicate that the clinical course of our 222 patients with idiopathic SHL showed no statistically significant differences from the clinical course of the patients with Lassa fever. We found a marked difference in age, however, and a clinically significant difference in the incidence of bilateral hearing loss. In reviewing the literature on sudden sensorineural hearing loss, we found no apparent relation between severity of viral illness and initial hearing loss or subsequent recovery. Cummins et al. suggest that virally induced hearing loss in Lassa fever is linked to the host's immune response and not to the viremia. We thus propose a virally induced immune response mechanism for idiopathic sensorineural SHL. Further prospective studies are needed for verification.

Cytoplasmic estrogen and progesterone receptors as prognostic parameters in primary endometrial carcinoma.

Eighty-six cases of primary endometrial carcinoma were assayed for the presence or absence of cytoplasmic estrogen and progesterone receptors by the saturation point dextran-coated charcoal assay. The levels of cytoplasmic progesterone receptors and estrogen receptors were analyzed according to clinical stage, histologic type and grade of the tumor, presence or absence of lymph node metastases, myometrial invasion, and survival. The cases were divided into positive and negative receptor groups with levels chosen of greater than 10 fmol/mg of cytosol protein for progesterone receptor and 5 fmol/mg of cytosol protein for estrogen receptor as discrimination points. Statistically significant survival differences were found between estrogen receptor positive versus estrogen receptor negative patients, progesterone receptor positive versus progesterone receptor negative patients, and estrogen positive-progesterone receptor positive versus estrogen negative-progesterone receptor negative patients. Mean cytoplasmic estrogen and progesterone receptor levels were inversely proportional to grade. This report suggests that treatment protocols should be devised to reflect the prognostic significance of receptor status.