Honokiol Exhibits Anti-NLRP3 Inflammasome and Antimicrobial Properties in Neisseria gonorrhoeae-Infected Macrophages.

Purpose: The NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome, crucial in infectious and inflammatory diseases by regulating IL-1ß, presents a target for disease management. Neisseria gonorrhoeae causes gonorrhea in over 87 million people annually, with previous research revealing NLRP3 inflammasome activation in infected macrophages. No natural products have been reported to counteract this activation. Exploring honokiol, a phenolic compound from Chinese herbal medicine, we investigated its impact on NLRP3 inflammasome activation in N. gonorrhoeae-infected macrophages. Methods: Honokiol's impact on the protein expression of pro-inflammatory mediators was analyzed using ELISA and Western blotting. The generation of intracellular H2O2 and mitochondrial reactive oxygen species (ROS) was detected through specific fluorescent probes (CM-H2DCFDA and MitoSOX, respectively) and analyzed by flow cytometry. Mitochondrial membrane integrity was assessed using specific fluorescent probes (MitoTracker and DiOC2(3)) and analyzed by flow cytometry. Additionally, the effect of honokiol on the viability of N. gonorrhoeae was examined through an in vitro colony-forming units assay. Results: Honokiol effectively inhibits caspase-1, caspase-11 and GSDMD activation and reduces the extracellular release of IL-1ß, NLRP3, and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) in N. gonorrhoeae-infected macrophages. Detailed investigations have demonstrated that honokiol lowers the production of H2O2 and the phosphorylation of ERK1/2 in N. gonorrhoeae-infected macrophages. Importantly, the phosphorylation of JNK1/2 and p38 and the activation of NF-κB remain unaffected. Moreover, honokiol reduces the N. gonorrhoeae-mediated generation of reactive oxygen species within the mitochondria, preserving their integrity. Additionally, honokiol suppresses the expression of the pro-inflammatory mediator IL-6 and inducible nitric oxide synthase induced by N. gonorrhoeae independently of NLRP3. Impressively, honokiol exhibits in vitro anti-gonococcal activity against N. gonorrhoeae. Conclusion: Honokiol inhibits the NLRP3 inflammasome in N. gonorrhoeae-infected macrophages and holds great promise for further development as an active ingredient in the prevention and treatment of symptoms associated with gonorrhea.

Cinnamaldehyde inhibits the NLRP3 inflammasome by preserving mitochondrial integrity and augmenting autophagy in Shigella sonnei-infected macrophages.

BACKGROUND: Worldwide, more than 125 million people are infected with Shigella each year and develop shigellosis. In our previous study, we provided evidence that Shigella sonnei infection triggers activation of the NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) inflammasome in macrophages. NLRP3 inflammasome is responsible for regulating the release of the proinflammatory cytokines interleukin (IL)-1ß and IL-18 through the protease caspase-1. Researchers and biotech companies have shown great interest in developing inhibitors of the NLRP3 inflammasome, recognizing it as a promising therapeutic target for several diseases. The leaves of Cinnamomum osmophloeum kaneh, an indigenous tree species in Taiwan, are rich in cinnamaldehyde (CA), a compound present in significant amounts. Our aim is to investigate how CA affects the activation of the NLRP3 inflammasome in S. sonnei-infected macrophages. METHODS: Macrophages were infected with S. sonnei, with or without CA. ELISA and Western blotting were employed to detect protein expression or phosphorylation levels. Flow cytometry was utilized to assess H2O2 production and mitochondrial damage. Fluorescent microscopy was used to detect cathepsin B activity and mitochondrial ROS production. Additionally, colony-forming units were employed to measure macrophage phagocytosis and bactericidal activity. RESULTS: CA inhibited the NLRP3 inflammasome in S. sonnei-infected macrophages by suppressing caspase-1 activation and reducing IL-1ß and IL-18 expression. CA also inhibited pyroptosis by decreasing caspase-11 and Gasdermin D activation. Mechanistically, CA reduced lysosomal damage and enhanced autophagy, while leaving mitochondrial damage, mitogen-activated protein kinase phosphorylation, and NF-κB activation unaffected. Furthermore, CA significantly boosted phagocytosis and the bactericidal activity of macrophages against S. sonnei, while reducing secretion of IL-6 and tumour necrosis factor following infection. CONCLUSION: CA shows promise as a nutraceutical for mitigating S. sonnei infection by diminishing inflammation and enhancing phagocytosis and the bactericidal activity of macrophages against S. sonnei.

The First-Water-Layer Evolution at the Graphene/Water Interface under Different Electro-Modulated Hydrophilic Conditions Observed by Suspended/Supported Field-Effect-Device Architectures.

Interfacial water molecules affect carrier transportation within graphene and related applications. Without proper tools, however, most of the previous works focus on simulation modeling rather than experimental validation. To overcome this obstacle, a series of graphene field-effect transistors (GFETs) with suspended (substrate-free, SF) and supported (oxide-supported, OS) configurations are developed to investigate the graphene-water interface under different hydrophilic conditions. With deionized water environments, in our experiments, the electrical transportation behaviors of the graphene mainly originate from the evolution of the interfacial water-molecule arrangement. Also, these current-voltage behaviors can be used to elucidate the first-water layer at the graphene-water interface. For SF-GFET, our experimental results show positive hysteresis in electrical transportation. These imply highly ordered interfacial water molecules with a separated-ionic distributed structure. For OS-GFET, on the contrary, the negative hysteresis shows the formation of the hydrogen-bond interaction between the interfacial water layer and the SiO2 substrate under the graphene. This interaction further promotes current conduction through the graphene/water interface. In addition, the net current-voltage relationship also indicates the energy required to change the orientation of the first-layer water molecules during electro-potential change. Therefore, our work gives an insight into graphene-water interfacial evolution with field-effect modulation. Furthermore, this experimental architecture also paves the way for investigating 2D solid-liquid interfacial features.

Poor Quality of Life in HIV-Infected Men Who Have Sex with Men is Associated with Excess-Type Constitution of Traditional Chinese Medicine.

Introduction: This study explored the pathological constitution as it relates to low quality of life in HIV-infected MSM patients, as a reference for clinical treatment. Methods: It had a cross-sectional research design using structured questionnaires to collect data, including patient's basic data, CD4+, CD4+/CD8+ ratio, Wang Qi constitution, and WHOQOL-BREF-Taiwan version questionnaires. We considered the association between constitutions and quality of life of HIV-infected MSM patients. Results and Discussion. The project accepted 203 HIV-infected MSM participants. The three most common pathological constitutions were Yang deficiency 15.5%, yin deficiency 13.1%, and qi deficiency 11.2%. The study determined scores for various quality of life domains: psychological (13.44 ± 2.27), social relationship (13.81 ± 2.80), physiological (14.43 ± 2.41), and environmental (14.78 ± 2.21). The TCM constitution is strongly correlated with the quality of life. Excess constitution had the worst quality of life. Comparing the infected time over one year with the time of <0-2 weeks, the adjusted odds ratios (AOR) were determined for abnormal CD4+ and CD4+/CD8+ ratio (OR: odds ratio: 0.03, 0.07, respectively, p < 0.001). Compared with the Gentleness constitution, there is a significant difference between the Deficiency and Excess constitution in sleep status and negative mood by multiple regression analysis (p < 0.001). Conclusion: The Excess constitutions was correlated with worse quality of life. Even if the immune system was restored, the psychosocial domain, sleep status, and negative mood were not improved.

Association of depression and antidepressant therapy with antiretroviral therapy adherence and health-related quality of life in men who have sex with men.

UNAIDS' HIV treatment targets require that 90% of people living with HIV/AIDS (PLWHA) receiving antiretroviral treatment (ART) achieve viral suppression and 90% of people with viral suppression have good health-related quality of life (HRQOL). This study aimed to examine the association of depression and antidepressant therapy with ART adherence and HRQOL in HIV-infected men who have sex with men (MSM). From 2018 through 2020, HIV-infected MSMs were consecutively recruited (N = 565) for the evaluation of ART adherence and HRQOL at Taipei City Hospital HIV clinics. Non-adherence to ART was defined as a Medication Adherence Report Scale score of < 23. HRQOL in PLWHHA was evaluated using WHOQOL-BREF, Taiwan version. Overall, 14.0% had depression and 12.4% exhibited non-adherence to ART. The nonadherence proportion was 21.8% and 10.5% in depressed and nondepressed HIV-infected MSM, respectively. After adjusting for other covariates, depression was associated with a higher risk of nonadherence to ART (adjusted odds ratio = 2.02; 95% confidence interval: 1.02-4.00). Physical, psychological, social, and environmental HRQOL were significantly negatively associated with depression. Considering antidepressant therapy, ART nonadherence was significantly associated with depression without antidepressant therapy but not with antidepressant therapy. The depressed HIV-infected MSM without antidepressant therapy had worse psychological, social, and environmental HRQOL than those with antidepressant therapy. Our study suggests that depression is associated with poor ART adherence and HRQOL, particularly in those without antidepressant therapy. Adequate diagnosis and treatment of depression should be provided for PLWHA to improve their ART adherence and HRQOL.

Intelligent Brushing Monitoring Using a Smart Toothbrush with Recurrent Probabilistic Neural Network.

Smart toothbrushes equipped with inertial sensors are emerging as high-tech oral health products in personalized health care. The real-time signal processing of nine-axis inertial sensing and toothbrush posture recognition requires high computational resources. This paper proposes a recurrent probabilistic neural network (RPNN) for toothbrush posture recognition that demonstrates the advantages of low computational resources as a requirement, along with high recognition accuracy and efficiency. The RPNN model is trained for toothbrush posture recognition and brushing position and then monitors the correctness and integrity of the Bass Brushing Technique. Compared to conventional deep learning models, the recognition accuracy of RPNN is 99.08% in our experiments, which is 16.2% higher than that of the Convolutional Neural Network (CNN) and 21.21% higher than the Long Short-Term Memory (LSTM) model. The model we used can greatly reduce the computing power of hardware devices, and thus, our system can be used directly on smartphones.

Critical Role for the NLRP3 Inflammasome in Mediating IL-1ß Production in Shigella sonnei-Infected Macrophages.

Shigella is one of the leading bacterial causes of diarrhea worldwide, affecting more than 165 million people annually. Among the serotypes of Shigella, Shigella sonnei is physiologically unique and endemic in human immunodeficiency virus-infected men who have sex with men. The NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) inflammasome, a protein complex composed of NLRP3, apoptosis-associated speck-like protein, and caspase-1, recognizes, and responds to pathogen infection and diverse sterile host-derived or environmental danger signals to induce IL-1ß and IL-18 production. Although the Shigella flexneri-mediated activation of the NLRP3 inflammasome has been reported, the effect of S. sonnei on NLRP3 inflammasome activation remains unclear. We found that S. sonnei induced IL-1ß production through NLRP3-dependent pathways in lipopolysaccharide-primed macrophages. A mechanistic study revealed that S. sonnei induced IL-1ß production through P2X7 receptor-mediated potassium efflux, reactive oxygen species generation, lysosomal acidification, and mitochondrial damage. In addition, the phagocytosis of viable S. sonnei was important for IL-1ß production. Furthermore, we demonstrated that NLRP3 negatively regulated phagocytosis and the bactericidal activity of macrophages against S. sonnei. These findings provide mechanistic insight into the activation of the NLRP3 inflammasome by S. sonnei in macrophages.

Methamphetamine Use Associated with Non-adherence to Antiretroviral Treatment in Men Who Have Sex with Men.

Methamphetamine is a prevalent recreational drug among men who have sex with men (MSM) living with HIV and could cause the cognitive impairment and memory loss. However, studies on the association between methamphetamine use and adherence to antiretroviral treatment (ART) are limited and had inconsistent findings. This study aimed to determine the impact of methamphetamine use on adherence to ART among MSM living with HIV. From December 2018 to October 2019, MSM living with HIV were recruited (N = 351) and non-adherence to ART was defined as a Medication Adherence Report Scale score of <23. Overall, 16.0% of the participants reported methamphetamine use in the prior three months and 13.4% of the participants had non-adherence to ART. The proportion of non-adherence to ART among HIV-positive MSM were 28.6% and 10.5% with and without methamphetamine use, respectively. After controlling for demographics, illicit drug use, and co-morbidities, methamphetamine use during the prior three months was associated with a higher risk of non-adherence to ART (adjusted odds ratio = 3.08; 95% confidence intervals: 1.24-7.69). Compared with HIV-positive MSM with non-adherence to ART, HIV-positive MSM with good adherence to ART had a higher CD4 counts and were more likely to achieve an undetectable viral load. Since poor adherence to ART is associated with an increased HIV viral load and the risk of HIV transmission to others, our study suggests that it is imperative to screen HIV-positive patients for methamphetamine use and to provide effective therapy to reduce methamphetamine use and the associated non-adherence to ART.

Whole-body vibration training improves balance control and sit-to-stand performance among middle-aged and older adults: a pilot randomized controlled trial.

BACKGROUND: Aging is associated with decreased balance, which increases falling risk. The objective of the current study was to determine the feasibility and effects of whole-body vibration (WBV) training on knee extensor muscle power, limits of stability, and sit-to-stand performance among community-dwelling middle-aged and older adults in the United States. METHODS: A randomized pilot study with participant blinding was conducted. Feasibility outcomes included recruitment and compliance rate. Twenty-nine community-dwelling older adults were randomly assigned to perform body-weight exercises with either an individualized vibration frequency and amplitude, a fixed vibration frequency and amplitude, or no vibration. Isokinetic knee extensor power, limits of stability, and sit-to-stand tests were conducted before beginning the exercises (baseline) and after 8 weeks of training. RESULTS: With a favorable recruitment rate (58%) and compliance rates (attrition 9%; adherence 85%), the intervention was deemed feasible. The limits of stability endpoint excursion score for the individualized frequency-amplitude group was increased by 8.8 (12.9%; P = 0.025) after training, and that group's maximum excursion score was increased by 9.2 (11.5%; P = 0.006) after training. The average weight transfer time score was significantly decreased by 0.2 s in the fixed group. The participants in the individualized group demonstrated a significant increase (3.2%) in weight rising index score after 8 weeks of WBV training. CONCLUSIONS: WBV training is feasible for use with elderly people, and this study achieved good recruitment and compliance. The present paper suggests that 8 weeks of WBV training improves limits of stability and sit-to-stand performance. Future studies must determine whether WBV training improves other factors that affect posture control. TRIAL REGISTRATION: This study was registered at the Texas Woman's University Institutional Review Board [TWU IRB 17632] on the 3rd of November 2014.

Hinokitiol induces DNA damage and autophagy followed by cell cycle arrest and senescence in gefitinib-resistant lung adenocarcinoma cells.

Despite good initial responses, drug resistance and disease recurrence remain major issues for lung adenocarcinoma patients with epidermal growth factor receptor (EGFR) mutations taking EGFR-tyrosine kinase inhibitors (TKI). To discover new strategies to overcome this issue, we investigated 40 essential oils from plants indigenous to Taiwan as alternative treatments for a wide range of illnesses. Here, we found that hinokitiol, a natural monoterpenoid from the heartwood of Calocedrus formosana, exhibited potent anticancer effects. In this study, we demonstrated that hinokitiol inhibited the proliferation and colony formation ability of lung adenocarcinoma cells as well as the EGFR-TKI-resistant lines PC9-IR and H1975. Transcriptomic analysis and pathway prediction algorithms indicated that the main implicated pathways included DNA damage, autophagy, and cell cycle. Further investigations confirmed that in lung cancer cells, hinokitiol inhibited cell proliferation by inducing the p53-independent DNA damage response, autophagy (not apoptosis), S-phase cell cycle arrest, and senescence. Furthermore, hinokitiol inhibited the growth of xenograft tumors in association with DNA damage and autophagy but exhibited fewer effects on lung stromal fibroblasts. In summary, we demonstrated novel mechanisms by which hinokitiol, an essential oil extract, acted as a promising anticancer agent to overcome EGFR-TKI resistance in lung cancer cells via inducing DNA damage, autophagy, cell cycle arrest, and senescence in vitro and in vivo.

Non-cytotoxic nanomaterials enhance antimicrobial activities of cefmetazole against multidrug-resistant Neisseria gonorrhoeae.

The emergence and spread of antibiotic-resistant Neisseria gonorrhoeae has led to difficulties in treating patients, and novel strategies to prevent and treat this infection are urgently needed. Here, we examined 21 different nanomaterials for their potential activity against N. gonorrhoeae (ATCC 49226). Silver nanoparticles (Ag NPs, 120 nm) showed the greatest potency for reducing N. gonorrhoeae colony formation (MIC: 12.5 µg/ml) and possessed the dominant influence on the antibacterial activity with their properties of the nanoparticles within a concentration range that did not induce cytotoxicity in human fibroblasts or epithelial cells. Electron microscopy revealed that the Ag NPs significantly reduced bacterial cell membrane integrity. Furthermore, the use of clinical isolates of multidrug-resistant N. gonorrhoeae showed that combined treatment with 120 nm Ag NPs and cefmetazole produced additive effects. This is the first report to screen the effectiveness of nanomaterials against N. gonorrhoeae, and our results indicate that 120 nm Ag NPs deliver low levels of toxicity to human epithelial cells and could be used as an adjuvant with antibiotic therapy, either for topical use or as a coating for biomaterials, to prevent or treat multidrug-resistant N. gonorrhoeae.

Automated flow system for sildenafil enrichment using surfactant coated solid-phase with fluorescence detection.

In this work, Amberlite XAD-1180 resin is used for on-line surfactant-mediated pre-concentration of sildenafil as a prior step for its fluorescent detection. In order to activate the column for sildenafil pre-concentration, the cationic surfactant (hexadecyltrimethylammoniunm bromide, HTAB) is adsorbed onto the resin. In these conditions, sildenafil is retained by HTAB-resin and then it is eluted with ethanol and analyzed by spectrofluorimetry. Drug-surfactant association produces a considerable fluorescence enhancement, increasing considerably the sensitivity of detection. Therefore, sildenafil can be pre-concentrated and quantitatively determined, with a detection limit of 0.2 ng mL(-1). The proposed method was successfully applied to the analysis of bulk drug, human urine, tablets, and local herbal medicine. Validation processes were performed by recovering studies and statistical analysis with satisfactory results.

Flow injection spectrofluorimetric determination of carvedilol mediated by micelles.

A novel flow injection (FI)-spectrofluorimetric methodology for the determination of carvedilol in microheterogeneous medium has been developed. In the sodium dodecyl sulfate (SDS) surfactant medium, an additional fluorescence enhancement was produced by the electrolyte NaCl. A total enhancement of 3.1-fold in the native fluorescent response was achieved respect to aqueous medium. Using an excitation and emission wavelength of 286 and 341 nm, respectively, a good linear relationship was obtained in the range of 9x10(-8) to 1x10(-6) mol L(-1) with a detection limit of 3.63x10(-9) mol L(-1) (S/N=3). This method was applied to determine carvedilol in commercial pharmaceutical formulations. Good concordance was found between the nominal (6.25, 12.5 and 25.0 mg) and experimental values. The new methodology developed showed high selectivity respect to the common excipients used in pharmaceuticals. The sampling rate was 30 samples h(-1). From the fluorescent properties, binding constant for carvedilol-SDS determined was 3.2x10(2) L mol(-1).

Invasive Haemophilus influenzae disease in adults in Taiwan.

BACKGROUND AND PURPOSE: Haemophilus influenzae is an important cause of invasive infection in infants and children, but it has been considered an uncommon cause of invasive disease in adults. We conducted a retrospective survey of invasive H. influenzae disease in adults in order to better understand the characteristics of clinical presentation and microbiology. METHODS: Patients older than 18 years with H. influenzae isolated from normally sterile sites, between July 1999 and June 2002 in a teaching hospital for adult patients were retrospectively analyzed. Data on demographics, clinical presentation, serotype, antibiotic susceptibility, and beta-lactamase production of H. influenzae isolates were analyzed. RESULTS: Fifteen patients were enrolled. The infectious diagnosis of invasive diseases comprised: pneumonia (5 patients), empyema (2), pelvic inflammatory disease (2), peritonitis (2), periorbital cellulitis with abscess formation (2), endophthalmitis (1) and primary bacteremia (1). Most patients were elderly with underlying illness. Of ten H. influenzae isolates available for analysis, two were serotype b and eight were nontypeable. Beta-lactamase production and ampicillin resistance were found in 6 H. influenzae isolates (5 nontypeable, and 1 type b). CONCLUSION: These data show H. influenzae disease in adults to be rare in Taiwan. Our limited number of cases suggest that nontypeable strains predominate in patients with invasive infection due to H. influenzae. Most patients had respiratory tract infections. Ampicillin resistance was found in more than one-half of H. influenzae isolates, and should be taken into consideration when antibiotics are prescribed on an empirical basis.

Clinical significance of Blastocystis hominis: experience from a medical center in northern Taiwan.

BACKGROUND AND PURPOSE: Blastocystis hominis is an intestinal protozoan. The pathogenic role of this organism in human beings is still controversial and has varied among reports from different geographic areas. The purpose of this study was to determine the clinical significance of B. hominis in northern Taiwan. METHODS: A total of 100 patients who had a positive B. hominis stool examination during the period April to December of 2001 were retrospectively identified from Taipei Veterans General Hospital. The demographic and clinical characteristics of these patients were reviewed from the medical records. RESULTS: All of the patients were adults. Fifty nine patients had more than one underlying diseases, including malignancies. Twenty one patients presented with fever and 10 patients had gastrointestinal symptoms, including diarrhea and/or abdominal pain. However, all of the patients had other conditions that might have contributed to the clinical presentation, and they improved without specific treatment for B. hominis. Furthermore, there were no significant differences in clinical symptoms and white blood cell count between patients with malignancy or diabetes mellitus and those without. Six patients had hypereosinophilia that could not be attributed to other causes. Among 34 patients who had a further stool examination within one year, B. hominis was undetectable in 31 patients (91.2%), despite their having no specific antiprotozoal treatment. CONCLUSIONS: The association of clinical symptoms and B. hominis could not be delineated from our study, even in immunocompromised patients. All of the patients improved without receiving any specific therapy. More studies from different areas are needed in order to delineate the clinical significance B. hominis.

On-line micellar-enhanced spectrofluorimetric determination of rhodamine dye in cosmetics.

A simple FI-fluorimetric analytical methodology for the continuous and sequential determination of rhodamine B (RhB) in cosmetic products has been developed and evaluated in terms of sensibility and selectivity. The influence of several surfactant solutions on RhB fluorescence signal has been studied; particular attention was paid in the aggregation behavior of RhB-SDS system. Linear response has been obtained in the range of 1.6 x 10(-9) and 1 x 10(-6) mol L(-1), with a detection limit of 5 x 10(-10) mol L(-1). The novel technique provides a simple dissolution of sample, on-line filtration with sampling rate higher than 100 samples h(-1) and has been satisfactorily applied to the RhB determination in commercial lipsticks.

Risk factors for mortality in patients with Acinetobacter baumannii bloodstream infection with genotypic species identification.

BACKGROUND AND PURPOSE: Acinetobacter baumannii is an increasingly common nosocomial infection with a high mortality rate. Identification of predictor factors of mortality from A. baumannii infection is important for the implementation of therapeutic management for patients with higher risk. However, many studies have reported data for Acinetobacter calcoaceticus-A. baumannii complex, which might lead to an uncertainty of results. In this study, we aimed to identify the predictive factors for mortality of patients infected with true A. baumannii that had been precisely identified by genotypic methodology. METHODS: Sixty seven patients with documented A. baumannii bacteremia were identified from a medical center in northern Taiwan during the period between February 1998 and February 2001. The patients' medical records were retrospectively reviewed. RESULTS: The risk factors associated with mortality in patients with A. baumannii bacteremia were underlying disease with malignancy, end-stage renal disease, and inappropriate antibiotic therapy. Laboratory variables, such as creatinine level, were also associated with poor prognosis by multivariate analysis. CONCLUSIONS: Increased serum creatinine level, malignancy and inappropriate therapy within 3 days were related to increased mortality in patients with A. baumannii bloodstream infection. Physicians should be aware of patients with poor prognostic factors and initiate prompt strategies, including appropriate antimicrobial therapy, in order to reduce mortality.

Adult candidemia at a medical center in northern Taiwan: a retrospective study.

BACKGROUND AND PURPOSE: Candidemia has been associated with a very high mortality. This study evaluated the predictors of candidemia-related mortality at a teaching hospital in northern Taiwan. METHODS: We conducted a retrospective analysis of adult patients with candidemia between September 2003 and May 2005. A stepwise logistic regression analysis was performed to determine the predictors of candidemia-associated mortality. All Candida isolates were identified to species by use of the ATB ID 32C kit and their susceptibilities to antifungal agents were tested by ATB Fungus 2 system. RESULTS: 179 episodes in 174 adult patients with candidemia were identified retrospectively. The predictors of mortality included duration of prior antibiotics >or=28 days, Acute Physiology and Chronic Health Evaluation (APACHE) II score >or=23 and retention of central venous catheters (CVCs). There was no statistically significant association between the time to the start of antifungal therapy and mortality from nosocomial candidemia. In addition, there was no significant association between the time to CVC removal and mortality after stratification by APACHE II score. CONCLUSIONS: Despite effective antifungal therapy after the onset of candidemia in more than half of the patients studied, mortality remained very high, especially in the groups with longer duration of prior antibiotic treatment, higher APACHE II score and CVC retention. Timing of CVC removal after onset of candidemia was not correlated with mortality.

Involvement of MAPKs and NF-kappaB in LPS-induced VCAM-1 expression in human tracheal smooth muscle cells.

Lipopolysaccharide (LPS) has been shown to induce the expression of adhesion molecules on airway epithelial and smooth cells and contributes to inflammatory responses. Here, the roles of mitogen-activated protein kinases (MAPKs) and nuclear factor-kappaB (NF-kappaB) pathways for LPS-induced vascular cell adhesion molecule (VCAM)-1 expression were investigated in HTSMCs. LPS-induced expression of VCAM-1 protein and mRNA in a time-dependent manner, was significantly inhibited by inhibitors of MEK1/2 (U0126), p38 (SB202190), and c-Jun-N-terminal kinase (JNK; SP600125). The involvement of p42/p44 MAPK and p38 in these responses was further confirmed by that transfection with small interference RNAs (siRNA) direct against MEK, p42, and p38 significantly attenuated LPS-induced VCAM-1 expression. Consistently, LPS-stimulated phosphorylation of p42/p44 MAPK and p38 was attenuated by pretreatment with U0126 or SB202190, and transfection with these siRNAs, respectively. In addition, LPS-induced VCAM-1 expression was significantly blocked by a specific NF-kappaB inhibitor helenalin. LPS-stimulated translocation of NF-kappaB into the nucleus and degradation of IkappaB-alpha was blocked by helenalin, U0126, SB202190, or SP600125. Moreover, the resultant enhancement of VCAM-1 expression increased the adhesion of polymorphonuclear cells to monolayer of HTSMCs which was blocked by pretreatment with helenalin, U0126, or SP600125 prior to LPS exposure. Taken together, these results suggest that in HTSMCs, activation of p42/p44 MAPK, p38, and JNK pathways, at least in part, mediated through NF-kappaB, is essential for LPS-induced VCAM-1 gene expression. These results provide new insight into the mechanisms of LPS action that bacterial toxins may promote inflammatory responses in the airway disease.

Determination of terazosin by cloud point extraction-fluorimetric combined methodology.

A new sensitive and selective preconcentration-fluorimetric method for determination of terazosin based on its native fluorescence was developed. The analyte, initially present in aqueous matrix, was treated with an extractive non-ionic surfactant solution and separated by the clouding phenomenon. The optimum analytical conditions for terazosin assay were established. Under these conditions, linear calibration curves were obtained over the range of 1x10(-5) to 7.0 microg mL(-1) with detection and quantification limits of 1.11x10(-5) and 3.7x10(-5)microg mL(-1), respectively. Additionally, the binding constant (K(B)) for the terazosin-PONPE 7.5 system was determined given a value of 1028 L mol(-1). The developed coupled methodology, which thoroughly satisfies the typical requirements for pharmaceutical control processes, was proved to be appropriate for monitoring terazosin in actual pharmaceutical formulations and biological fluid sample. The results were validated by recovery test and by comparison with other reported methods, being highly satisfactory.