Capsaicin exerts therapeutic effects by targeting tNOX-SIRT1 axis and augmenting ROS-dependent autophagy in melanoma cancer cells.

Although considered a sporadic type of skin cancer, malignant melanoma has regularly increased internationally and is a major cause of cancer-associated death worldwide. The treatment options for malignant melanoma are very limited. Accumulating data suggest that the natural compound, capsaicin, exhibits preferential anticancer properties to act as a nutraceutical agent. Here, we explored the underlying molecular events involved in the inhibitory effect of capsaicin on melanoma growth. The cellular thermal shift assay (CETSA), isothermal dose-response fingerprint curves (ITDRFCETSA), and CETSA-pulse proteolysis were utilized to confirm the direct binding of capsaicin with the tumor-associated NADH oxidase, tNOX (ENOX2) in melanoma cells. We also assessed the cellular impact of capsaicin-targeting of tNOX on A375 cells by flow cytometry and protein analysis. The essential role of tNOX in tumor- and melanoma-growth limiting abilities of capsaicin was evaluated in C57BL/6 mice. Our data show that capsaicin directly engaged with cellular tNOX to inhibit its enzymatic activity and enhance protein degradation capacity. The inhibition of tNOX by capsaicin was accompanied by the attenuation of SIRT1, a NAD+-dependent deacetylase. The suppression of tNOX and SIRT1 then enhanced ULK1 acetylation and induced ROS-dependent autophagy in melanoma cells. Capsaicin treatment of mice implanted with melanoma cancer cells suppressed tumor growth by down-regulating tNOX and SIRT1, which was also seen in an in vivo xenograft study with tNOX-depleted melanoma cells. Taken together, our findings suggest that tNOX expression is important for the growth of melanoma cancer cells both in vitro and in vivo, and that inhibition of the tNOX-SIRT1 axis contributes to inducting ROS-dependent autophagy in melanoma cells.

Identify the interactions between phytochemicals and proteins in the complicated food matrix.

Phytochemicals usually mix with food proteins in our regular diet. Unexpected interactions may lead to changes in bioaccessibility, bioactivity, and bioavailability of phytochemicals. However, our understanding of these interactions between phytochemical and food proteins is limited because of the experimental restrictions. Here, we used pulse-proteolysis to conduct the unfolding equilibrium and dose-dependent experiments on the food proteins for the first time. The interaction between epigallocatechin gallate (EGCG) and caseins was identified in the complicated food matrix, whole milk. Another food proteome, soymilk, was also optimized for identifying the binding targets of EGCG and caffeine. Among the identified interactions, the mixing of milk with coffee generates the most prominent masking effect of 46.61 ± 3.86% relative to the calculated antioxidant capacity. Our results demonstrated that pulse proteolysis is applicable for identifying the interactions between phytochemicals and proteins in the complicated food matrix.

Capsaicin acts through tNOX (ENOX2) to induce autophagic apoptosis in p53-mutated HSC-3 cells but autophagy in p53-functional SAS oral cancer cells.

Despite the progress that has been made in diagnosing and treating oral cancers, they continue to have a poor prognosis, with a 5-year overall survival rate of approximately 50%. We have intensively studied the anticancer properties of capsaicin (a burning constituent of chili pepper), mainly focusing on its apoptotic properties. Here, we investigated the interplay between apoptosis and autophagy in capsaicin-treated oral cancer cells with either functional or mutant p53. Cytotoxicity was determined by cell impedance measurements and WST-1 assays, and cell death was analyzed by flow cytometry. The interaction between capsaicin and tumor-associated NADH oxidase (tNOX, ENOX2) was studied by cellular thermal shift assay (CETSA) and isothermal dose-response fingerprint curves (ITDRFCETSA). Our CETSA data suggested that capsaicin directly engaged with tNOX, resulting in its degradation through the ubiquitin-proteasome and the autophagy-lysosome systems. In p53-functional SAS cells, capsaicin induced significant cytotoxicity via autophagy but not apoptosis. Given that tNOX catalyzes the oxidation of NADH, the direct binding of capsaicin to tNOX also inhibited the NAD+-dependent activity of sirtuin 1 (SIRT1) deacetylase, we found that capsaicin-induced autophagy involved enhanced acetylation of ULK1, which is a key player in autophagy activation, possibly through SIRT1 inhibition. In p53-mutated HSC-3 cells, capsaicin triggered both autophagy and apoptosis. In this case, autophagy occurred before apoptosis: during this early stage, autophagy seemed to inhibit apoptosis; at a later stage, in contrast, autophagy appeared to be essential for the induction of apoptosis. Western blot analysis revealed that the reduction in tNOX and SIRT1 associated with enhanced ULK1 acetylation and c-Myc acetylation, which in turn, reactivated the TRAIL pathway, ultimately leading to apoptosis. Taken together, our data highlight the potential value of leveraging capsaicin and tNOX in therapeutic strategies against oral cancer.

Metabolites modulate the functional state of human uridine phosphorylase I.

Metabolic pathways in cancer cells typically become reprogrammed to support unconstrained proliferation. These abnormal metabolic states are often accompanied by accumulation of high concentrations of ATP in the cytosol, a phenomenon known as the Warburg Effect. However, how high concentrations of ATP relate to the functional state of proteins is poorly understood. Here, we comprehensively studied the influence of ATP levels on the functional state of the human enzyme, uridine phosphorylase I (hUP1), which is responsible for activating the chemotherapeutic pro-drug, 5-fluorouracil. We found that elevated levels of ATP decrease the stability of hUP1, leading to the loss of its proper folding and function. We further showed that the concentration of hUP1 exerts a critical influence on this ATP-induced destabilizing effect. In addition, we found that ATP interacts with hUP1 through a partially unfolded state and accelerates the rate of hUP1 unfolding. Interestingly, some structurally similar metabolites showed similar destabilization effects on hUP1. Our findings suggest that metabolites can alter the folding and function of a human protein, hUP1, through protein destabilization. This phenomenon may be relevant in studying the functions of proteins that exist in the specific metabolic environment of a cancer cell.

D-methionine improves cisplatin-induced anorexia and dyspepsia syndrome by attenuating intestinal tryptophan hydroxylase 1 activity and increasing plasma leptin concentration.

BACKGROUND: Cisplatin is a widely used antineoplastic drug. However, cisplatin-induced dyspepsia syndromes, including delayed gastric emptying, gastric distension, early satiety, nausea, and vomiting, often force patients to take doses lower than those prescribed or even refuse treatment. D-methionine has an appetite-enhancing effect and alleviates weight loss during cisplatin treatment. METHODS: This work established a model of anorexia and dyspepsia symptoms with intraperitoneal injection of cisplatin (5 mg/kg) once a week for three cycles. Presupplementation with or without D-methionine (300 mg/kg) was performed. Orexigenic and anorexigenic hormones (ghrelin, leptin, and glucagon-like peptide-1), tryptophan hydroxylase 1 (TPH1), 5-hydroxytryptamine receptors (5-HT2C and 5-HT3 ), and hypothalamic feeding-related peptides were measured by immunohistochemistry staining, enzyme-linked immunosorbent assay, and real-time PCR assay. KEY RESULTS: Cisplatin administration caused marked decrease in appetite and body weight, promoted adipose and fat tissue atrophy, and delayed gastric emptying and gastric distension, and D-methionine preadministration prior to cisplatin administration significantly ameliorated these side effects. Besides, cisplatin induced an evident increase in serum ghrelin level, TPH1 activity, and 5-HT3 receptor expression in the intestine and decreased plasma leptin levels and gastric ghrelin mRNA gene expression levels. D-methionine supplementation recovered these changes. The expression of orexigenic neuropeptide Y/agouti-related peptide and anorexigenic cocaine- and amphetamine-regulated transcript proopiomelanocortin neurons were altered by D-methionine supplementation in cisplatin-induced anorexia rats. CONCLUSIONS AND INFERENCES: D-methionine supplementation prevents cisplatin-induced anorexia and dyspepsia syndrome possibly by attenuating intestinal tryptophan hydroxylase 1 activity and increasing plasma leptin concentration. Therefore, D-methionine can be used as an adjuvant therapy for treating cisplatin-induced adverse effects.

Immune response evoked by tumor-associated NADH oxidase (tNOX) confers potential inhibitory effect on lung carcinoma in a mouse model.

Tumor-associated NADH oxidase (tNOX, ENOX2), which belongs to a family of growth-related NADH oxidases, was originally identified as a plasma membrane protein of rat hepatoma and is inhibited or downregulated by several anti-cancer drugs. The objective of this study was to evaluate the anti-tumor effects of tNOX used as an immunogen against Lewis lung cancer. Human tNOX was expressed in Escherichia coli, purified by His-Tag affinity chromatography, and emulsified with the adjuvant, ISA 201 VG. Immunological analyses of the generated tNOX vaccine were performed in mice. The results of ELISA and ELISpot were significantly higher in tNOX vaccine group compared to the control group. In vivo, we examined the anti-tumor effects of mice that received the tNOX vaccine via the intraperitoneal or subcutaneous routes. Mice were vaccinated three times at 2-week intervals, challenged at 2 weeks after the final vaccination, and terminated at 34 days post-challenge. Antibody titers, tumor volume and histopathological scores were used to evaluate the anti-tumor effects of the tNOX vaccine. Our results revealed that tNOX-vaccinated mice had significantly higher antibody titers than negative control (NC) and challenge control (CC) mice. When compared to the corresponding CC groups, the intraperitoneal and subcutaneous vaccination with tNOX showed a significantly smaller tumor mass volume (P < 0.05) and a significantly lower histological lesion score (P < 0.05), respectively. Our results demonstrate that the use of a xenogeneic tNOX as an immunogen in mice activates immune responses and anti-tumor effects against Lewis lung cancer.

A one-pot analysis approach to simplify measurements of protein stability and folding kinetics.

To achieve a good understanding of the characteristics of a protein, it is important to study its stability and folding kinetics. Investigations of protein stability have been recently applied to drug-target identification, drug screening, and proteomic studies. The efficiency of the experiments performed to study protein stability and folding kinetics is now a crucial factor that needs to be optimized for these potential applications. However, the standard procedures used to carry out these experiments are usually complicated and time consuming. Large number of measurements is the bottleneck that limits the application of protein folding to large-scale experiments. To overcome this limitation, we developed a method denoted as "one-pot analysis" which is based on taking a single measurement from a mixture of samples rather than from every sample. We combined one-pot analysis with pulse proteolysis to determine the effects of the binding of maltose to maltose-binding protein on the protein folding properties. After carrying out a simple optimization, we demonstrated that protein stability or unfolding kinetics could be measured accurately with just one detection measurement. We then further applied the optimized conditions to cellular thermal shift assay (CETSA). Combining one-pot analysis with CETSA led to a successful determination of the effects of the binding of methotrexate to dihydrofolate reductase in HCT116 cancer cells. Our results demonstrated the applicability of one-pot analysis to energetics-based methods for studying protein folding. We expect the combination of one-pot analysis and energetics-based methods to significantly benefit studies such as drug-target identification, proteomic investigations, and drug screening.

Regulation of tNOX expression through the ROS-p53-POU3F2 axis contributes to cellular responses against oxaliplatin in human colon cancer cells.

BACKGROUND: Oxaliplatin belongs to the platinum-based drug family and has shown promise in treating cancer by binding to DNA to induce cytotoxicity. However, individual patients show diverse therapeutic responses toward oxaliplatin due to yet-unknown underlying mechanisms. We recently established that oxaliplatin also exert its anti-cancer activity in gastric cancer cell lines by targeting tumor-associated NADH oxidase (tNOX), attenuate NAD+ generation and reduce NAD+-dependent sirtuin 1 (SIRT1) deacetylase activity, which in turn enhances p53 acetylation and apoptosis. METHODS: In this study, differential cellular outcomes in response to oxaliplatin exposure of p53-wild-type versus p53-null HCT116 human colon cancer cells were examined. Cell growth profile was determined by cell impedance measurements and apoptosis was analyzed by flow cytometry. The engagement between oxaliplatin and tNOX protein was studied by cellular thermal shift assay. Furthermore, western blot analysis revealed that p53 was important in regulating tNOX expression in these cell lines. RESULTS: In p53-wild-type cells, we found that oxaliplatin inhibited cell growth by inducing apoptosis and concurrently down-regulating tNOX at both the transcriptional and translational levels. In p53-null cells, in contrast, oxaliplatin moderately up-regulated tNOX expression and yielded no apoptosis and much less cytotoxicity. Further experiments revealed that in p53-wild-type cells, oxaliplatin enhanced ROS generation and p53 transcriptional activation, leading to down-regulation of the transcriptional factor, POU3F2, which enhances the expression of tNOX. Moreover, the addition of a ROS scavenger reversed the p53 activation, POU3F2 down-regulation, and apoptosis induced by oxaliplatin in p53-wild-type cells. In the p53-null line, on the other hand, oxaliplatin treatment triggered less ROS generation and no p53 protein, such that POU3F2 and tNOX were not down-regulated and oxaliplatin-mediated cytotoxicity was attenuated. CONCLUSION: Our results show that oxaliplatin mediates differential cellular responses in colon cancer cells depending on their p53 status, and demonstrate that the ROS-p53 axis is important for regulating POU3F2 and its downstream target, tNOX. Notably, the depletion of tNOX sensitizes p53-null cells to both spontaneous and oxaliplatin-induced apoptosis. Our work thus clearly shows a scenario in which targeting of tNOX may be a potential strategy for cancer therapy in a p53-inactivated system.

ATP alters protein folding and function of Escherichia coli uridine phosphorylase.

Uridine phosphorylase is one of the critical enzymes in the pyrimidine salvage pathway. Cells regenerate uridine for nucleotide metabolism by incorporating uracil with ribose-1-phosphate with this enzyme. Recent studies indicate that Escherichia coli uridine phosphorylase is destabilized in the presence of ATP. However, the mechanism underlying the destabilization process and its influence on uridine phosphorylase function remain to be established. Here, we comprehensively investigated the effects of ATP on protein folding and function of Escherichia coli uridine phosphorylase. Our results demonstrate that ATP apparently decreases the stability of uridine phosphorylase in a concentration-dependent manner. Additionally, simply increasing the level of ATP led to a reduction of enzymatic activity to complete inhibition. Further studies showed that uridine phosphorylase accumulates as a partially unfolded state in the presence of ATP. Moreover, ATP specifically accelerated the unfolding rate of uridine phosphorylase with no observable effects on the refolding process. Our preliminary findings suggest that ATP can alter the protein folding and function of enzymes via apparent destabilization. This mechanism may be significant for proteins functioning under conditions of high levels of ATP, such as cancer cell environments.

Structure of a partially unfolded form of Escherichia coli dihydrofolate reductase provides insight into its folding pathway.

Proteins frequently fold via folding intermediates that correspond to local minima on the conformational energy landscape. Probing the structure of the partially unfolded forms in equilibrium under native conditions can provide insight into the properties of folding intermediates. To elucidate the structures of folding intermediates of Escherichia coli dihydrofolate reductase (DHFR), we investigated transient partial unfolding of DHFR under native conditions. We probed the structure of a high-energy conformation susceptible to proteolysis (cleavable form) using native-state proteolysis. The free energy for unfolding to the cleavable form is clearly less than that for global unfolding. The dependence of the free energy on urea concentration (m-value) also confirmed that the cleavable form is a partially unfolded form. By assessing the effect of mutations on the stability of the partially unfolded form, we found that native contacts in a hydrophobic cluster formed by the F-G and Met-20 loops on one face of the central ß-sheet are mostly lost in the partially unfolded form. Also, the folded region of the partially unfolded form is likely to have some degree of structural heterogeneity. The structure of the partially unfolded form is fully consistent with spectroscopic properties of the near-native kinetic intermediate observed in previous folding studies of DHFR. The findings suggest that the last step of the folding of DHFR involves organization in the structure of two large loops, the F-G and Met-20 loops, which is coupled with compaction of the rest of the protein.

Rapid screening for sickle cell disease by polymerase chain reaction-high resolution melting analysis.

Each year, ~300,000 individuals with sickle cell disease (SCD), a hemoglobinopathy caused by ß-globin gene mutation, are born, and >75% of those are in Africa. The present study examined 511 individuals on the island of Bioko (Equatorial Guinea) and attempted to establish a method for rapid sickle cell disease screening. Following DNA extraction and polymerase chain reaction (PCR) amplification, high resolution melting (HRM) analysis was used to assess the specificity of fluorescence signals of the PCR products and to differentiate various genotypes of these products. The analytical results of HRM were validated using DNA sequencing. By HRM analysis, 80 out of 511 samples were classified as hemoglobin S (Hb S) heterozygotes, while 431 out of 511 samples were classified as wild-type. No mutant homozygote was identified. DNA sequencing indicated that within the 431 wild-type samples as indicated by HRM analysis, one case was actually a Hb S heterozygote and another case was a rare hemoglobin S-C genotype (sickle-hemoglobin C disease). One out of 80 suspected Hb S heterozygotes as indicated by HRM was confirmed as wild-type by DNA sequencing and the results of residual 508 cases were consistent for HRM analysis and sequencing. In conclusion, HRM analysis is a simple, high-efficiency approach for Hb S screening and is useful for early diagnosis of SCD and particularly suitable for application in the African area.

Andrographis paniculata Extract and Andrographolide Modulate the Hepatic Drug Metabolism System and Plasma Tolbutamide Concentrations in Rats.

Andrographolide is the most abundant terpenoid of A. paniculata which is used in the treatment of diabetes. In this study, we investigated the effects of A. paniculata extract (APE) and andrographolide on the expression of drug-metabolizing enzymes in rat liver and determined whether modulation of these enzymes changed the pharmacokinetics of tolbutamide. Rats were intragastrically dosed with 2 g/kg/day APE or 50 mg/kg/day andrographolide for 5 days before a dose of 20 mg/kg tolbutamide was given. APE and andrographolide reduced the AUC0-12 h of tolbutamide by 37% and 18%, respectively, compared with that in controls. The protein and mRNA levels and enzyme activities of CYP2C6/11, CYP1A1/2, and CYP3A1/2 were increased by APE and andrographolide. To evaluate whether APE or andrographolide affected the hypoglycemic action of tolbutamide, high-fat diet-induced obese mice were used and treated in the same manner as the rats. APE and andrographolide increased CYP2C6/11 expression and decreased plasma tolbutamide levels. In a glucose tolerance test, however, the hypoglycemic effect of tolbutamide was not changed by APE or andrographolide. These results suggest that APE and andrographolide accelerate the metabolism rate of tolbutamide through increased expression and activity of drug-metabolizing enzymes. APE and andrographolide, however, do not impair the hypoglycemic effect of tolbutamide.

The beneficial effect of ST-36 (Zusanli) acupressure on postoperative gastrointestinal function in patients with colorectal cancer.

PURPOSE/OBJECTIVES: To evaluate the effectiveness of ST-36 (Zusanli) acupressure on recovery of postoperative gastrointestinal function in patients with colorectal cancer. DESIGN: A longitudinal, randomized, controlled trial design. SETTING: An urban medical center in Taiwan. SAMPLE: 60 patients with colorectal cancer who had undergone abdominal surgery. METHODS: Patients were randomly assigned to two groups, the ST-36 acupressure group (n = 30) and a sham acupressure group (n = 30). Patients in the ST-36 group received an acupressure procedure in a three-minute cycle performed three times per day during the five days after surgery. Patients in the control group received routine postoperative care and sham acupressure. Generalized estimating equations (GEEs) were used to gauge longitudinal effects of the two groups of patients. MAIN RESEARCH VARIABLES: Frequency of bowel sounds, the time to first flatus passage, first liquid intake, solid intake, and defecation. FINDINGS: Patients who received acupressure had significantly earlier flatus passage and time to liquid intake as compared to patients in the control group. Other main variables, including the first time to solid intake and defecation, did not show significant difference between the two groups. The GEE method revealed that all patients had increasing bowel sounds over time, and the experimental group had greater improvement of bowel motility than the control group within the period of 2-3 days postoperatively. CONCLUSIONS: ST-36 acupressure was able to shorten the time to first flatus passage, oral liquid intake, and improve gastrointestinal function in patients after abdominal surgery. IMPLICATIONS FOR NURSING: ST-36 acupressure can be integrated into postoperative adjunct nursing care to assist patients' postoperative gastrointestinal function. KNOWLEDGE TRANSLATION: Few studies have explored the effectiveness of acupressure techniques on promoting bowel sounds. Evidence from this study suggests stimulation of the ST-36 acupressure point can increase bowel sound frequency for patients with colorectal cancer in the first three days after surgery. Application of this technique may improve a patient's comfort after surgery.

[Reducing occupational burnout and enhancing job performance in new nurses: the efficacy of "last mile" programs].

BACKGROUND: New nurses undergo a stressful and challenging transition process in the nursing workplace. Lack of patient care knowledge and skills and work adaption difficulties lead to a high turnover rate that drains essential new talent away from the nursing profession and further exacerbates professional staffing shortages in the healthcare sector. The "last mile" program is a program developed jointly by a nursing school and hospital as a mechanism to bridge classroom learning to clinical practice and smooth the transition of nursing students into nursing professionals. PURPOSE: The purpose of this study was to understand the effect of the "last mile" program on job performance and occupational burnout among new nurses. METHODS: We conducted a quasi-experimental study in 2009 on a convenience sample of new nurses in a medical center. Participants were assigned into two groups, namely those enrolled in the last mile program (n = 29) and those not enrolled in the program (n = 94). Research team members and several collaborative universities developed the last mile program used in this study; Seven experts established content validity; The last mile program included 84 hours of lecture courses and 160 hours of clinical practice. Data was collected using the nursing job performance scale developed in 2007 by Greenslade and Jimmieson and translated ÷ back translated into an equivalent Chinese version. Exploratory factor analysis showed all items aggraded into 8 factors, which could be divided into task performance and contextual performance concept categories. Task performance concepts included: social support, information, coordination of care, and technical care; Contextual performance concepts included: interpersonal support, job-task support, volunteering for additional duties and compliance. The Cronbach's α for the 8 factors were .70-.95. The occupational burnout inventory included the 4 subscales of personal burnout, work-related burnout, client-related burnout, and over-commitment, with associated Cronbach's α ranging from .84-.90. Data was collected at one, three, and six months after employment. Repeated measures ANOVA and an independent t-test were used to analyze data. RESULTS: The average age of the 123 participants surveyed was 23 years, with no differences identified between last-mile and non-last-mile groups in terms of education level, work unit, or other demographic variables. New nurses who participated in the last mile program achieved significantly higher performance scores for job-task support, volunteering for additional duties, and overall task and contextual performance than those who did not. Last-mile-program group participants also had significantly lower client-related burnout than their non-last-mile-program peers. CONCLUSIONS / IMPLICATIONS FOR PRACTICE: The last mile program facilitates new nurses' contextual performance and reduces incidence of care burnout. The cooperative education model linking universities and hospitals can be a positive component in a new nurse retention strategy for hospital administrators and educators.

Selective stabilization of a partially unfolded protein by a metabolite.

When proteins fold in vivo, the intermediates that exist transiently on their folding pathways are exposed to the potential interactions with a plethora of metabolites within the cell. However, these potential interactions are commonly ignored. Here, we report a case in which a ubiquitous metabolite interacts selectively with a nonnative conformation of a protein and facilitates protein folding and unfolding process. From our previous proteomics study, we have discovered that Escherichia coli glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which is not known to bind ATP under native conditions, is apparently destabilized in the presence of a physiological concentration of ATP. To decipher the origin of this surprising effect, we investigated the thermodynamics and kinetics of folding and unfolding of GAPDH in the presence of ATP. Equilibrium unfolding of the protein in urea showed that a partially unfolded equilibrium intermediate accumulates in the presence of ATP. This intermediate has a quaternary structure distinct from the native protein. Also, ATP significantly accelerates the unfolding of GAPDH by selectively stabilizing a transition state that is distinct from the native state of the protein. Moreover, ATP also significantly accelerates the folding of GAPDH. These results demonstrate that ATP interacts specifically with a partially unfolded form of GAPDH and affects the kinetics of folding and unfolding of this protein. This unusual effect of ATP on the folding of GAPDH implies that endogenous metabolites may facilitate protein folding in vivo by interacting with partially unfolded intermediates.

Energetics-based discovery of protein-ligand interactions on a proteomic scale.

Biochemical functions of proteins in cells frequently involve interactions with various ligands. Proteomic methods for the identification of proteins that interact with specific ligands such as metabolites, signaling molecules, and drugs are valuable in investigating the regulatory mechanisms of cellular metabolism, annotating proteins with unknown functions, and elucidating pharmacological mechanisms. Here we report an energetics-based target identification method in which target proteins in a cell lysate are identified by exploiting the effect of ligand binding on their stabilities. Urea-induced unfolding of proteins in cell lysates is probed by a short pulse of proteolysis, and the effect of a ligand on the amount of folded protein remaining is monitored on a proteomic scale. As proof of principle, we identified proteins that interact with ATP in the Escherichia coli proteome. Literature and database mining confirmed that a majority of the identified proteins are indeed ATP-binding proteins. Four identified proteins that were previously not known to interact with ATP were cloned and expressed to validate the result. Except for one protein, the effects of ATP on urea-induced unfolding were confirmed. Analyses of the protein sequences and structure models were also employed to predict potential ATP binding sites in the identified proteins. Our results demonstrate that this energetics-based target identification approach is a facile method to identify proteins that interact with specific ligands on a proteomic scale.

Awareness of hypertension guidelines in Taiwanese nurses: a questionnaire survey.

BACKGROUND AND RESEARCH OBJECTIVE: Nurses play an important role in hypertension prevention and management because of their unique positions in patient education. However, the effectiveness of patient education relies largely on the nurse's level of awareness of the current hypertension guidelines. The purpose of this study was to examine the level of awareness of hypertension guidelines and associated factors among nurses in Taiwan. SUBJECTS AND METHODS: A cross-sectional survey was conducted in 10 hospitals in northern Taiwan. The Hypertension Management Questionnaire was developed based on the Taiwan Hypertension Guidelines and the seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure. RESULTS: The survey return rate was 95.9%. A total of 1418 nurses were included in the analysis. Adequate guideline awareness was found in 49.5% of the total sample. Among the 7 dimensions of the Hypertension Management Questionnaire, the definition of hypertension, methods for blood pressure measurements, and impact of high blood pressure on cardiovascular disease had the lowest rates of correct answers. Multiple regression analysis revealed that the nurses' clinical experience, educational level, work setting, in-service education training on hypertension, and level of the hospital (R2 = 35.4%, F = 52.89, P < .001) independently predicted the nurse's level of awareness. CONCLUSIONS: A large proportion of the nurses in northern Taiwan had insufficient knowledge of the hypertension guidelines.

Risk factors associated with unplanned endotracheal self-extubation of hospitalized intubated patients: a 3-year retrospective case-control study.

This 3-year retrospective case-control study aimed to identify risk factors associated with unplanned endotracheal self-extubation (UESE) of hospitalized intubated patients and to compare unplanned and planned extubation groups' characteristics of patients and nurses, vital signs, serum laboratory values, Glasgow Coma Scale scores, Acute Physiology and Chronic Health Evaluation II (APACHE II) scores, and use of physical restraints and sedatives. The study found that most UESEs occurred during evening or night shifts or during shifts staffed by nurses with less experience and less education. Most of the self-extubated patients (80%) were physically restrained. Pulse rate and APACHE II score were both significant predictors of UESE. Efforts to prevent UESEs should include identification of patients at higher risk.

[Role transition and working adaption in new nursing graduates: a qualitative study].

BACKGROUND: The role transition process is full of stresses and challenges for nurses. Between 35-61% nurses leave their job within the first year. Past cross-sectional quantitative studies have not provided deep descriptions of either the dynamic role transition or work adaption processes of new nurses. PURPOSE: The purpose of this study was to understand the role transition experience of new nurses as they transitioned into clinical practice during their first three months on the job. METHODS: A qualitative approach was used. Data were collected through a semi-structured interview from 50 new nurses. Data were analyzed using category-content analysis. RESULTS: Three stages were identified in the new nurse work adaption process over the first three-month period. These included (1) Understanding: New nurse knowledge and skills are insufficient to handle routine work, adapting to the role transition is difficult, feelings of anxiety emerge related to fears of incompetence, communication difficulties must be faced in the handover process, new nurses adopt feelings of attachment to their preceptors, they must work to adopt appropriate attitudes and approaches to nursing practice, and support is sought from family, teachers and friends; (2) Acclimation: Learning to care for patients independently, seeking role models, learning to adapt to night shifts, trying to identify with co-workers, and seeking support from colleagues, preceptors and head nurses; (3) Acceptance: Managing nursing work better in terms of time and organization, feeling gradual acceptance from co-workers, restoring personal enthusiasm for work, starting to consider other, non-work related matters, experiencing and appreciating the support of co-workers and head nurses. CONCLUSIONS/IMPLICATION: New nurses face a critical role transition process through their first three months on the job. Guidance and leadership from experienced nurses and multiple support systems can assist new nurses to acclimate to their role. Research results provide information for educators and administrators to better understand the adaption process of new nurses, and offer a reference for developing future strategies to improve nurse competency in handling their work.

Comparison of family partnership intervention care vs. conventional care in adult patients with poorly controlled type 2 diabetes in a community hospital: a randomized controlled trial.

BACKGROUND: Although there is a significant correlation between the degree of family support and clinical outcome, little research has focused on the effectiveness of family partnership intervention care (FPIC) for patients with poorly controlled type 2 diabetes. OBJECTIVES: This study aimed to compare FPIC with conventional care (CC) across a number of outcome measures in patients with poorly controlled type 2 diabetes. DESIGN: The study was performed using a randomized controlled trial design. PARTICIPANTS: Patients with poorly controlled type 2 diabetes who were solely treated with oral antidiabetic agents and had at least two out of three hemoglobin A1C readings equal to or above 7% in the previous 12 months, were randomly assigned to the FPIC group (n=28) and to the CC group (n=28). METHODS: Baseline and 6-month follow-up scores were compared using the following outcome measures: (1) hemoglobin A1C, (2) BMI, (3) lipid profile, (4) family supportive behaviours, (5), knowledge of and attitudes toward diabetes, and (6) diabetes self-care behaviours. Descriptive and non-parametric statistics were employed to compare differences in outcome measures between the groups. RESULTS: There were no significant differences in the reduction of hemoglobin A1C levels (p=0.46), lipid profile values (p>0.05), and improvement of diabetes self-care behaviours (p=0.61) between the groups at 6 months post-intervention. However, there were significant differences in the scores of family supportive behaviours (p=0.031) and patients' knowledge of and attitudes toward diabetes between the groups (p<0.05). CONCLUSION: These findings support the use of FPIC to enhance family supportive behaviours, and to improve patients' knowledge of and attitudes toward diabetes. Thus, the study is of value in helping policy decision-makers to develop more effective diabetes control intervention programmes.