Mass spectrometric identification of phenol-soluble modulins in the ATCC® 43300 standard strain of methicillin-resistant Staphylococcus aureus harboring two distinct phenotypes.

Staphylococcus aureus subsp. aureus Rosenbach (ATCC® 43300™) is a representative methicillin-resistant S. aureus (MRSA) strain that is used as a quality control strain for testing the methicillin susceptibility of clinical isolates. It has been consistently observed that colonies with two different colors (golden yellow and white) grow from the ATCC stock on blood agar plates. In this study, staphylococcal peptide and protein profiling was performed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. From the extract of the potentially hemolytic 43300 strain, we identified a single significant peak at an observed mass-to-charge (m/z) value of 2306.9, which correlates well with the predicted mass of formylated phenol-soluble modulin α2, a major staphylococcal virulence factor. Subsequent liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis revealed that the hemolytic 43300 cells predominantly produced various phenol-soluble modulins (PSMs) and their export proteins, including four α type PSM peptides, PSMß1, PSM-mec, PmtC and PmtD. However, none of the PSM peptides was detected in the presumably non-hemolytic 43300 strain, but the export proteins PmtC and PmtD. We found that this MRSA standard strain expresses two distinct phenotypes and that their phenotypic features are more likely associated with PSM toxin production. Therefore, careful attention is needed when MRSA reference strains are utilized for the diagnosis and susceptibility testing of MRSA in clinical settings.

Effect of polyacetylenic acids from Prunella vulgaris on various plant pathogens.

AIMS: This study is aiming at characterizing antifungal substances from the methanol extract of Prunella vulgaris and at investigating those substances' antifungal and antioomycete activities against various plant pathogens. METHODS AND RESULTS: Two polyacetylenic acids were isolated from P. vulgaris as active principles and identified as octadeca-9,11,13-triynoic acid and trans-octadec-13-ene-9,11-diynoic acid. These two compounds inhibited the growth of Magnaporthe oryzae, Rhizoctonia solani, Phytophthora infestans, Sclerotinia sclerotiorum, Fusarium oxysporum f. sp. raphani, and Phytophthora capsici. In addition, these two compounds and the wettable powder-type formulation of an n-hexane fraction of P. vulgaris significantly suppressed the development of rice blast, tomato late blight, wheat leaf rust, and red pepper anthracnose. CONCLUSIONS: These data show that the extract of P. vulgaris and two polyacetylenic acids possess antifungal and antioomycete activities against a broad spectrum of tested plant pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the occurrence of octadeca-9,11,13-triynoic acid and trans-octadec-13-ene-9,11-diynoic acid in P. vulgaris and their efficacy against plant diseases. The crude extract containing the two polyacetylenic acids can be used as a natural fungicide for the control of various plant diseases.

Mycofumigation with Oxyporus latemarginatus EF069 for control of postharvest apple decay and Rhizoctonia root rot on moth orchid.

AIMS: To characterize the volatile antifungal compound produced by Oxyporus latemarginatus EF069 and to examine in vitro and in vivo fumigation activity of the fungus. METHODS AND RESULTS: An antifungal volatile-producing strain, O. latemarginatus EF069 inhibited the mycelial growth of Alternaria alternata, Botrytis cinerea, Colletotrichum gloeosporioides, Fusarium oxysporum f. sp. lycopersici, and Rhizoctonia solani by mycofumigation. An antifungal volatile compound was isolated from the hexane extract of wheat bran-rice hull cultures of O. latemarginatus EF069 by repeated silica gel column chromatography and identified as 5-pentyl-2-furaldehyde (PTF). The purified PTF inhibited mycelial growth of R. solani in a dose-dependent manner. The mycofumigation with solid cultures of EF069 also reduced effectively the development of postharvest apple decay caused by B. cinerea and Rhizoctonia root rot of moth orchid caused by R. solani. CONCLUSIONS: Oxyporus latemarginatus EF069 showed in vitro and in vivo fumigation activity against plant pathogenic fungi by producing 5-pentyl-2-furaldehyde. SIGNIFICANCE AND IMPACT OF THE STUDY: Oxyporus latemarginatus EF069 producing an antifungal volatile compound may be used as a biofumigant for the control of fungal plant diseases.

Effects of neolignans from the stem bark of Magnolia obovata on plant pathogenic fungi.

AIMS: To characterize antifungal principles from the methanol extract of Magnolia obovata and to evaluate their antifungal activities against various plant pathogenic fungi. METHODS AND RESULTS: Four neolignans were isolated from stem bark of M. obovata as antifungal principles and identified as magnolol, honokiol, 4-methoxyhonokiol and obovatol. In mycelial growth inhibition assay, both magnolol and honokiol displayed more potent antifungal activity than 4-methoxyhonokiol and obovatol. Both magnolol and honokiol showed similar in vivo antifungal spectrum against seven plant diseases tested; both compounds effectively suppressed the development of rice blast, tomato late blight, wheat leaf rust and red pepper anthracnose. 4-Methoxyhonokiol and obovatol were highly active to only rice blast and wheat leaf rust respectively. CONCLUSIONS: The extract of M. obovata and four neolignans had potent in vivo antifungal activities against plant pathogenic fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: Neolignans from Magnolia spp. can be used and suggested as a novel antifungal lead compound for the development of new fungicide and directly as a natural fungicide for the control of plant diseases such as rice blast and wheat leaf rust.

Bikaverin and fusaric acid from Fusarium oxysporum show antioomycete activity against Phytophthora infestans.

AIMS: To isolate and identify antioomycete substances from Fusarium oxysporum EF119 against Phytophthora infestans and to investigate their antimicrobial activities against various plant pathogenic bacteria, oomycetes and true fungi. METHODS AND RESULTS: Two antioomycete substances were isolated from liquid cultures of F. oxysporum EF119, which shows a potent disease control efficacy against tomato late blight caused by P. infestans. They were identified as bikaverin and fusaric acid by mass and nuclear magnetic resonance spectral analyses. They inhibited the mycelial growth of plant pathogenic oomycetes and fungi. Fusaric acid also effectively suppressed the cell growth of various plant pathogenic bacteria, but bikaverin was virtually inactive. Treatment with bikaverin at 300 microg ml(-1) suppressed the development of tomato late blight by 71%. Fusaric acid provided effective control against tomato late blight and wheat leaf rust over 67% at concentrations more than 100 microg ml(-1). CONCLUSIONS: Both bikaverin and fusaric acid showed in vitro and in vivo antioomycete activity against P. infestans. SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium oxysporum EF119 producing both bikaverin and fusaric acid may be used as a biocontrol agent against tomato late blight caused by P. infestans.

Chronic deep brain stimulation of subthalamic and anterior thalamic nuclei for controlling refractory partial epilepsy.

OBJECTIVES: Experimental data and case reports of intractable epilepsy patients treated with deep brain stimulation (DBS) of the internal nuclei suggest a considerable anticonvulsant effect. We intended to describe the results of DBS on subthalamic nuclei and anterior thalamic nuclei (STN and ATN) from our patients and to evaluate the long-term efficiency and safety of DBS for controlling intractable epilepsy. METHODS: Six patients with refractory epilepsy and inadequate for surgery were implanted with DBS electrodes (3 in STN and 3 in ATN, respectively), switched on after a week of insertion followed by chronological observation. Seizure counts were monitored and compared with pre-implantation baseline. RESULTS: There was significant clinical improvement in respect of reduction of seizure frequency as well as the alleviation of ictal severity in almost patients. The mean reduction in seizure frequency was 62.3% (49.1% from STN vs. 75.4% from ATN). Except one patient (patient 3) with accidental infection on the right anterior chest, no complication or withdrawal of DBS was seen during our study. CONCLUSION: DBS on STN and ATN demonstrated their clear efficiency and relative safety comparable or superior to previous studies during long term follow-up. Subsequent, well designed studies warrant the further increase of the knowledge about antiepileptic effect of DBS.

Evaluation of 2-[18F]-fluoro-2-deoxy-D-glucose positron emission tomography/computed tomography in rat models with hepatocellular carcinoma with liver cirrhosis.

Liver cirrhosis is a predominant risk factor for hepatocellular carcinoma (HCC). However, the exact mechanism of the progression from cirrhosis to cancer remains unclear. The uptake of 2-[(18)F]-fluoro-2-deoxy-D-glucose ((18)F-FDG) is widely used as a marker of increased glucose metabolism to monitor the progression of cancer with positron emission tomography (PET)/computed tomography (CT). Here we investigated the feasibility of using (18)F-FDG PET/CT in the diethylnitrosamine (DEN) mediated experimental hepatocellular carcinoma model. Rats received weekly intraperitoneal injections of DEN for 16 weeks for induction of HCC. We recorded starting from 0 days or 0 weeks after the last DEN injection. The weight and survival rate of rats were then measured. Also, an (18)F-FDG PET scan and serum analysis were performed at minus 2, 0, plus 2, and plus 4 weeks after the last DEN injection. The body weight of rats was maintained between 350 g and 370 g during 14 and 20 weeks, and the rats were euthanized at 35 days after the last DEN injection. The serum levels of alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphate (ALP) were significantly higher at zero weeks after the last DEN injection. The (18)F-FDG uptake for the quantitative evaluation of HCC was done by measuring the region of interest (ROI). At minus two weeks after the last DEN injection, the ROI of rats had significantly increased compared to the normal group, in a time-dependent manner. These results suggest that FDG uptake serves as a good screening test to evaluate the feasibility of DEN-induced HCC.

Use of laser Doppler flowmetry for estimation of the depth of burns.

The trend for treatment of deep second degree burns and third degree burns is toward early excision and skin grafting. The ability to predict burn depth accurately as early as possible is important for early excision and skin grafting. This study, prospectively evaluated the ability of laser Doppler flow measurements, obtained within 72 hours after burn injury, to predict the depth of burn wounds. A Periflux system 4001 laser Doppler flowmeter was used to measure the cutaneous microflow circulation of 100 selected points of burn wounds on 44 inpatients and of 1680 selected points on 120 volunteers from March of 1993 to February of 1994. The mean value of superficial second degree burns checked by laser Doppler was 194.6 perfusion units (PU). The value of deep second degree burns was 59.7 PU, and the value of third degree burns was 5.1 PU. The mean normal cutaneous blood flow of 120 volunteers (control group) was between 4 and 9 PU, except on the head, neck, hand, and foot. Blood flow of more than 100 PU correctly predicted (90.2 percent of cases) a superficial second degree burn. Blood flow between 100 and 10 PU correctly predicted (96.2 percent of cases) a deep second degree dermal burn. That of less than 10 PU correctly predicted (100 percent of cases) a third degree burn. There was also a significant correlation between initial flow measurements and the depth of burn wounds. We conclude that laser Doppler flow measurements performed early after burn injury are useful in predicting the depth of burn wounds. Laser Doppler flowmetry has the advantage of being easy to use and noninvasive and of providing immediate results for early determination of burn depth. Laser Doppler flowmetry is useful in selecting patients for early excision and grafting of burn wounds.

Loss of Mel-18 induces tumor angiogenesis through enhancing the activity and expression of HIF-1α mediated by the PTEN/PI3K/Akt pathway.

Mel-18 has been implicated in several processes in tumor progression, in which the Akt pathway is involved as an important key molecular event. However, the function of Mel-18 in human cancers has not been fully established yet. Here, we examined the effect of Mel-18 on tumor angiogenesis in human breast cancer, and found that Mel-18 was a novel regulator of HIF-1α. Mel-18 negatively regulated the HIF-1α expression and its target gene VEGF transcription during both normoxia and hypoxia. We demonstrated that Mel-18 regulated the HIF-1α expression and activity via the PI3K/Akt pathway. Loss of Mel-18 downregulated Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) expression, consequently activating the PI3K/Akt/MDM2 pathway, and leading to an increase of HIF-1α protein level. Mel-18 modulated the HIF-1α transcriptional activity via regulating the cytoplasmic retention of FOXO3a, a downstream effector of Akt, and recruitment of HIF-1α/CBP complex to the VEGF promoter. Furthermore, our data shows that Mel-18 blocked tumor angiogenesis both in vitro and in vivo. Mel-18 overexpression inhibited in vitro tube formation in human umbilical endothelial cells (HUVECs). Xenografts in NOD/SCID mice derived from stably Mel-18 knocked down MCF7 human breast cancer cells showed increased tumor volume, microvessel density, and phospho-Akt and HIF-1α expression levels. In conclusion, our findings provide that Mel-18 is a novel regulator of tumor angiogenesis through regulating HIF-1α and its target VEGF expressions mediated by the PTEN/PI3K/Akt pathway, suggesting a new tumor-suppressive role of Mel-18 in human breast cancer.

Novel tumor suppressive function of Smad4 in serum starvation-induced cell death through PAK1-PUMA pathway.

DPC4 (deleted in pancreatic cancer 4)/Smad4 is an essential factor in transforming growth factor (TGF)-ß signaling and is also known as a frequently mutated tumor suppressor gene in human pancreatic and colon cancer. However, considering the fact that TGF-ß can contribute to cancer progression through transcriptional target genes, such as Snail, MMPs, and epithelial-mesenchymal transition (EMT)-related genes, loss of Smad4 in human cancer would be required for obtaining the TGF-ß signaling-independent advantage, which should be essential for cancer cell survival. Here, we provide the evidences about novel role of Smad4, serum-deprivation-induced apoptosis. Elimination of serum can obviously increase the Smad4 expression and induces the cell death by p53-independent PUMA induction. Instead, Smad4-deficient cells show the resistance to serum starvation. Induced Smad4 suppresses the PAK1, which promotes the PUMA destabilization. We also found that Siah-1 and pVHL are involved in PAK1 destabilization and PUMA stabilization. In fact, Smad4-expressed cancer tissues not only show the elevated expression of PAK1, but also support our hypothesis that Smad4 induces PUMA-mediated cell death through PAK1 suppression. Our results strongly suggest that loss of Smad4 renders the resistance to serum-deprivation-induced cell death, which is the TGF-ß-independent tumor suppressive role of Smad4.

Some fungal endophytes from vegetable crops and their anti-oomycete activities against tomato late blight.

AIMS: To isolate endophytic fungi from vegetable plants and examine their in vivo anti-oomycete activity against Phytophthora infestans in tomato plants. METHODS AND RESULTS: Endophytic fungi were isolated from surface-sterilized plant tissues and anti-oomycete activity was measured by in vivo assay using tomato seedlings. Endophytic fungi showing potent anti-oomycete activity were identified by morphological characteristics and nuclear ribosomal ITS1-5.8S-ITS2 sequence analysis. A total of 152 isolates were obtained from 66 healthy tissue samples of cucumber, red pepper, tomato, pumpkin and Chinese cabbage and the fermentation broths of 23 isolates showed potent in vivo anti-oomycete activity against tomato late blight with control values over 90%. The Fusarium oxysporum strain EF119, which was isolated from roots of red pepper, showed the most potent disease control efficacy against tomato late blight. In dual-culture tests, it inhibited the growth of Pythium ultimum, P. infestans and Phytophthora capsici. CONCLUSIONS: Among endophytic fungi isolated from healthy tissues of vegetable plants, F. oxysporum EF119 showed the most potent in vivo anti-oomycete activity against tomato late blight and in vitro anti-oomycete activity against several oomycete pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Endophytic fungi showing anti-oomycete activity in vitro and in vivo may be used as biocontrol agents particularly of tomato late blight.

Comparison of oral sensory function in complete denture and implant-supported prosthesis wearers.

In order to compare the differences of part of the oral sensory functions among natural dentition, complete denture wearers and implant-supported prostheses wearers, tactile and pressure awareness was measured. Tactile sense was estimated by the thickness perception threshold between upper and lower dentition. Sensibility of pressure feeling was evaluated by threshold of lateral loading on tooth. Within this limited experiment, it could be concluded that an osseointegrated root form implant helped towards restoration of oral sensory functions.

Comparison of castability and surface roughness of commercially pure titanium and cobalt-chromium denture frameworks.

STATEMENT OF PROBLEM: Titanium is a biocompatible material, but it is not widely used in clinical dentistry for conventional removable denture frameworks. Little research exists on its applicability. PURPOSE: This study compared the casting accuracy and roughness of titanium and cobalt-chromium denture frameworks. MATERIAL AND METHODS: Twenty Kennedy Class II, Division 1 removable partial denture frameworks were fabricated with commercially pure titanium (n = 10) and a cobalt-chromium alloy (n = 10). The casting accuracy of each framework was determined by visual, radiographic, and microscopic methods. The roughness of each polished framework surface was analyzed with atomic force microscopy. RESULTS: The clinical fit, porosities, and microporosities of both types of metal frameworks were qualitatively similar. The surface roughness of polished pure titanium and cobalt-chromium frameworks was 104.43 +/- 69.24 nm and 133.91 +/- 40.92 nm, respectively. This difference was not statistically significant (P >.05). CONCLUSION: The clinical fit, porosity, and surface roughness of the titanium and cobalt-chromium frameworks fabricated for this study were comparable.

[The effect of patient teaching on compliance with sick role behavior of diabetic patients].

This study examined the effect of patient teaching on Compliance with sick role behavior in diabetic patients. The purpose was to improve diabetic patients Compliance by D.M. patient teaching. The study objectives were to determine the effect of patient teaching on Compliance with sick role behavior, and factors influencing compliance with sick role behavior of diabetic patients. The subjects, consisting of 52 diabetic patients diagnosed in the C. and other hospitals in K. city were divided into experimental and control groups. Data were gathered from July 25th to September 3rd, 1988 through interviews by questionnaires, measurement of blood sugar level by Reflux. D.M. Patient teaching was defined as informational intervention of social support by the nurse. A booklet representing patient education and questionnaires were developed by the investigator, and were tested for Content validity, and reliability by Item Analysis: Cronbachs alpha for any instrument to measure variables was patient Compliance .83, perceived health belief .65, diabetic knowledge .70. Analysis of data were done by paired t-test, t-test, Anova, Pearson correlation, and Stepwise multiple regression. The results of the study may be summarized as follows: 1. The effect of patient teaching on Compliance with sick role behavior, on diabetic Knowledge and health belief was Confirmed by significant differences between the experimental and the control group before and after the experiments. (P = 000 P = 006, P = 004).(ABSTRACT TRUNCATED AT 250 WORDS)

Mapping of the human body skin with laser Doppler flowmetry.

The purpose of this study was to evaluate cutaneous blood flow in the human body to better establish normal flow ranges and to determine whether the location, side of the body, sex, and age affect the flow range at seven different sites. From March 1993 to February 1994 a Periflux system 4001 laser Doppler flowmeter was used to measure the cutaneous microflow circulation of 1,680 selected points in 120 volunteers. The mean normal cutaneous blood flow of 120 volunteers was between 4 and 9 perfusion units (PU) except in the head, neck, hand, and foot. Mean cutaneous blood flow measurements were as follows: the upper arm, 6.6 +/- 1.20 PU (mean +/- SE); the forearm, 6.7 +/- 1.95 PU; the thorax, 7.1 +/- 1.72 PU; the flank, 6.3 +/- 1.23 PU; the abdomen, 5.3 +/- 1.79 PU; the thigh, 4.8 +/- 1.34 PU; the lower leg, 4.6 +/- 1.39 PU (p < 0.05). The blood flow of the thorax was highest and was 54% higher than the lower leg. The group of teenagers had the highest cutaneous blood flow, with an average value of 6.9 +/- 0.62 PU. The group of subjects in their sixties proved to have the lowest cutaneous blood flow-32% less than the teenagers. Gender differences were not noted. There were no significant differences in blood flow in regard to the side of the body.