Gaseous CO2 signal initiates growth of butyric-acid-producing Clostridium butyricum in both pure culture and mixed cultures with Lactobacillus brevis.
Microbial strains produce numerous volatile substances in the anaerobic conditions of the human intestines. The availability of CO(2) is known to be a prerequisite for bacterial growth in general. In experiments with anaerobic Lactobacillus brevis and Clostridium butyricum bacteria in the Portable Microbial Enrichment Unit (PMEU) it was shown that these strains interact; this interaction being mediated by CO(2) emission. CO(2) promoted clostridial growth in pure cultures and mixed cultures with lactobacilli. The growth of C. butyricum in pure cultures was much delayed or did not start at all without CO(2) from outside. Conversely, the onset of growth was provoked by a short (15 min) CO(2) burst. In mixed cultures the presence of lactobacilli in equal numbers speeded up the onset of clostridial growth by 10 h. If C. butyricum cultures designated as PMEU 1, 2, and 3 in cultivation syringes were chained by connecting the gas flow thereby allowing the volatiles of the preceding syringe culture to bubble to the next one, the growth started in 20, 10, or 6 h, respectively. This effect of gaseous emissions from other cultures speeding up the bacterial growth initiation was abolished if the gas was passed through sodium hydroxide to remove the CO(2). The positive contribution of lactobacilli to the growth of butyric-acid-producing clostridia documented in this simulation experiment with PMEU has in vivo implications and indicates molecular communication between the species. CO(2) is a necessary signal for the growth of clostridia, and lactobacilli can promote clostridial growth in mixed cultures where both bacteria grow well with mutual benefit.


The clinical nurse specialist as resuscitation process manager.
The purpose of this article was to describe the history and leadership dimensions of the role of resuscitation process manager and provide specific examples of how this role is implemented at a Midwest medical center. In 1992, a medical center in the Midwest needed a nurse to manage resuscitation care. This role designation meant that this nurse became central to all quality improvement efforts in resuscitation care. The role expanded as clinical resuscitation guidelines were updated and as the medical center grew. The role became known as the critical care clinical nurse specialist as resuscitation process manager. This clinical care nurse specialist was called a manager, but she had no direct line authority, so she accomplished her objectives by forming a multitude of collaborative networks. Based on a framework by Finkelman, the manager role incorporated specific leadership abilities in quality improvement: (1) coordination of medical center-wide resuscitation, (2) use of interprofessional teams, (3) integration of evidence into practice, and (4) staff coaching to develop leadership. The manager coordinates resuscitation care with the goals of prevention of arrests if possible, efficient and effective implementation of resuscitation protocols, high quality of patient and family support during and after the resuscitation event, and creation or revision of resuscitation policies for in-hospital and for ambulatory care areas. The manager designs a comprehensive set of meaningful and measurable process and outcome indicators with input from interprofessional teams. The manager engages staff in learning, reflecting on care given, and using the evidence base for resuscitation care. Finally, the manager role is a balance between leading quality improvement efforts and coaching staff to implement and sustain these quality improvement initiatives. Revisions to clinical guidelines for resuscitation care since the 1990s have resulted in medical centers developing improved resuscitation processes that require management. The manager enhances collaborative quality improvement efforts that are in line with Institute of Medicine recommendations. The role of resuscitation process manager may be of interest to medical centers striving for excellence in evidence-based resuscitation care.


Evaluation of toxicity after acute accidental methotrexate ingestions in children under 6 years old: a 16-year multi-center review.
There is little data on the frequency of adverse events following acute methotrexate ingestions in pediatric patients. Likewise, recommendations for observation length, site and management strategies in this population are not well established. Therefore, most recommendations are modeled after management of chronic overdose in patients with underlying medical conditions. The primary objective of this study is to determine the frequency of acute toxicity after acute methotrexate accidental unsupervised ingestions in patients less than six years. In addition, we describe the frequency of late toxicity and characterize the management site and approaches. This is a retrospective cohort study of pediatric accidental unsupervised methotrexate ingestions reported to six poison centers in the United States over a 16 year period. Demographic information, exposure details, signs, symptoms, treatments, length and location of observation and outcomes were collected. 103 patients met inclusion criteria. Methotrexate dose was reported in 86 patients (84%) and ranged from 1.3 mg-75 mg. The majority of cases (97%) ingested a dose ≤20 mg. The significant majority of cases experienced no clinical effects (99 of 103 cases; 96%). Three children experienced minor outcome (3%). There were no patients with a major outcome or death. The incidence of toxicity from pediatric single, acute ingestions of methotrexate is rare and when it occurs is generally limited to no or only minimally concerning effects. Because concentrations from single ingestions were consistent with low subtoxic exposures, we believe that home monitoring without hospital referral and without methotrexate specific therapy is reasonable in those with acute ingestions up to 20 mg.


Economics of antihypertensive therapy.
Antihypertensive therapy has come a long way in the past 40 years. With the wide range of drugs now available, cost has become an additional consideration in selection of therapy. However, precise figures for comparison of drugs within and between classes can be difficult and time-consuming to gather. The unit-cost tables in this article are presented with the hope that they will simplify the selection process. Of course, unit cost is not the sole criterion to be considered; potency and frequency of administration also influence the overall cost of therapy.


Quantification of γδ T cells and HLA-DR+ NK cells does not predict emergence of new contrast enhancing lesions in MS patients suspending natalizumab treatment.
Natalizumab (NTZ) is a drug that has been widely used in the treatment of multiple sclerosis (MS). NTZ is very effective in suppressing inflammation, but if treatment is suspended many patients will experience relapses. To investigate if quantification of γδ T cells and HLA-DR+ NK cells could predict early disease reactivation after NTZ suspension. Absolute counts of γδ T cells and HLA-DR+ NK cells in whole blood were determined with flow cytometry in fifteen patients treated with NTZ. NTZ treatment was then withdrawn and patients were followed with clinical visits and MR investigations. Patients with recurrent disease had higher absolute counts of γδ T cells 129 (±156) cells/μl in comparison to patients with stable disease 50.0 (±51.0) cells/μl but the difference was not statistically significant and largely driven by outliers. Patients with recurrent and stable disease had similar absolute counts of HLA-DR+ NK cells. Quantification of γδ T cells and HLA-DR+ NK cells could not predict active disease after NTZ suspension.


Mitochondria in oral cancer stem cells: Unraveling the potential drug targets for new and old drugs.
Head and neck cancer is a major health problem worldwide, with most cases arising in the oral cavity. Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer, accounting for over 90% of all cases. Compared to other types of cancer, OSCC, has the worse prognosis, with a 5-year survival rate of 50%. Additionally, OSCC is characterized by a high rate of resistance to chemotherapy treatment, which may be partly explained by the presence of cancer stem cells (CSC) subpopulation. CSC can adapt to harmful environmental condition and are highly resistant to both chemotherapy and radiotherapy treatments, thus contributing to tumor relapse. The aim of this review is to highlight the role of mitochondria in oral CSC as a potential target for oral cancer treatment. For this purpose, we reviewed some fundamental aspects of the most validated protein markers of stemness, autophagy, the mitochondrial function and energy metabolism in oral CSC. Moreover, a discussion will be made on why energy metabolism, especially oxidative phosphorylation in CSC, may offer such a diverse source of original pharmacological target for new drugs. Finally, we will describe some drugs able to disturb mitochondrial function, with emphasis on those aimed to interrupt the electron transport chain function, as novel therapeutic strategies in multidrug-resistant oral CSC. The reutilization of old drugs approved for clinical use as new antineoplastics, in cancer treatment, is also matter of revision.


Characteristic neuroradiologic features in hemorrhagic shock and encephalopathy syndrome.
Hemorrhagic shock and encephalopathy syndrome is a devastating disease, but the pathogenesis remains unclear. The aim of this study was to examine the usefulness of neuroimaging in establishing a diagnosis and elucidating the pathogenesis. We analyzed the neuroradiologic features of 22 patients who fulfilled the Levin criteria. All patients underwent brain computed tomography (CT), and 14 patients underwent brain magnetic resonance imaging (MRI) including diffusion-weighted imaging in 10 patients. Initial CT showed normal findings in 14 of 18 (78%) patients, but subsequently hypodensities appeared in bilateral watershed zones and progressed to whole brain edema. MRI revealed cytotoxic edema, showing hyperintensities in bilateral watershed zones on diffusion-weighted imaging with a low apparent diffusion coefficient. Serial neuroimaging showed characteristic features of a widespread brain ischemic event mainly in watershed zones in hemorrhagic shock and encephalopathy syndrome.


Behavioral treatments for adult dental avoidance. A stepped-care approach.
The growing research literature dealing with the psychologic treatment of dental fear and avoidance suggests several interventions as effective, but provides little guidance in choosing among them. Under these circumstances, experienced psychologic practitioners may choose among these interventions on the basis of their own clinical impressions as to which treatment might be best suited to each patient they see. An alternative approach is the stepped-care approach in which the least expensive/most practical intervention is implemented, with more costly or complex procedures implemented only if the less expensive intervention proves unsuccessful. Each of the behavioral treatments for dental fear that has received researchers' attention is briefly described, and they are ordered in terms of their costs in professional time. Finally, an account of the special precautions needed to implement a stepped-care strategy for the reduction of dental fear is outlined.


Induction of subaortic septal ischaemia to reduce obstruction in hypertrophic obstructive cardiomyopathy. Studies to develop a new catheter-based concept of treatment.
To develop a new catheter-based method of treatment in patients with hypertrophic obstructive cardiomyopathy. Does abolition of the blood supply to the subaortic part of the septum lead to regional myocardial ischaemia and a decrease in the left ventricular outflow tract gradient? To find this out, in 10 consecutive patients the first larger septal branch of the left anterior descending coronary artery was temporarily occluded with conventional percutaneous transluminal coronary angioplasty. The intracoronary electrocardiogram was registered for objective verification of the intended ischaemia. The intraventricular pressure was measured at rest and at the post extrasystolic beat under programmed electrostimulation of the right ventricle. During occlusion, regional ischaemia was observed in all patients. Simultaneously, there was a significant reduction of the intraventricular gradient from 56.2 mmHg to 32.2 mmHg (P < 0.05) followed by an increase from 32.2 mmHg to 61.1 mmHg (P < 0.01) after release of occlusion of the septal branch. During ischaemia there was no increase in left ventricular end-diastolic pressure. We conclude that the results form the basis for a new catheter interventional therapy in hypertrophic obstructive cardiomyopathy.


Growth status and menarche in urban and rural China.
To examine the relationship between current age at menarche and growth status in an urban and rural area of Eastern China. Cross-sectional survey: self-completion questionnaire and anthropometry in 12 schools in urban Hangzhou and rural Chunan in Zhejiang Province. The median menarcheal age calculated by probit analysis was significantly different in the two areas: 12.8 years (SD 0.9) in the urban area and 13.2 (SD 1.0) in the rural area (p < 0.001). Girls who reach menarche are significantly heavier and taller with higher BMIs than those of the same age who are pre-menarche. After adjustment for BMI and other possible confounders, urban girls were still menstruating significantly earlier than girls in rural areas (OR 3.3, 2.1-5.2). The age of menarche is probably still declining in China. Although BMI is an important factor in the onset of menstruation, some other unmeasured environmental variable may be implicated in this population.


Amyloid deposition and small vessel disease are associated with cognitive function in older adults with type 2 diabetes.
Diabetes is associated with cognitive decline, but the underlying mechanisms are complex and their relationship with Alzheimer's Disease biomarkers is not fully understood. We assessed the association of small vessel disease (SVD) and amyloid burden with cognitive functioning in 47 non-demented older adults with type-2 diabetes from the Israel Diabetes and Cognitive Decline Study (mean age 78Y, 64% females). FLAIR-MRI, Vizamyl amyloid-PET, and T1W-MRI quantified white matter hyperintensities as a measure of SVD, amyloid burden, and gray matter (GM) volume, respectively. Mean hemoglobin A1c levels and duration of type-2 diabetes were used as measures of diabetic control. Cholesterol level and blood pressure were used as measures of cardiovascular risk. A broad neuropsychological battery assessed cognition. Linear regression models revealed that both higher SVD and amyloid burden were associated with lower cognitive functioning. Additional adjustments for type-2 diabetes-related characteristics, GM volume, and cardiovascular risk did not alter the results. The association of amyloid with cognition remained unchanged after further adjustment for SVD. Our findings suggest that SVD and amyloid pathology may independently contribute to lower cognitive functioning in non-demented older adults with type-2 diabetes, supporting a multimodal approach for diagnosing, preventing, and treating cognitive decline in this population.


Effects of anti-resorptive drugs on implant survival and peri-implantitis in patients with existing osseointegrated dental implants: a retrospective cohort study.
The effect of anti-resorptive drug (ARD) usage among patients with successful dental implant osseointegration is controversial. This study showed an increased risk of implant failure in ARD users. Risk factors included pre-existing marginal bone loss, overdenture, diabetes, and a short interval between implant placement and ARD administration. This retrospective study aimed to determine whether anti-resorptive drug (ARD) usage increased risk of implant failure among patients with successful implant osseointegration. Additionally, the study investigated risk factors that affected implant survival rate in ARD users. Eighty ARD users with 344 implants who had more than 12 months of follow-up from the initiation of ARD treatment during the period between 2008 and 2017 were included, along with 80 non-ARD users from the same period. The primary outcome was dental implant survival. Kaplan-Meier survival curves and Cox proportional hazard models were used for survival analysis. Average follow-up was 85.3 months. Implant survival rates were 89.83% in ARD users and 96.03% in non-ARD users. In the univariate Cox proportional hazard model, risk of implant failure was significantly higher in patients with pre-existing marginal bone loss (MBL), diabetes, and concurrent bone augmentation. However, risk of implant failure was significantly lower when the interval between implant placement and ARD administration was < 36 months. Compared with overdenture, single crown and fixed splinted users had lower risk of implant failure. In multivariate analysis, variables including pre-existing MBL, diabetes, < 36-month interval between implant placement and ARD treatment, and usage of fixed splinted prosthesis were significantly associated with increased risk of implant failure. ARD administration after implant osseointegration was correlated with a reduced implant survival rate. Pre-existing MBL, diabetes, type of final prosthesis, and the interval between implant placement and initiation of ARD administration influenced risk of implant failure.


Intracytoplasmic lumina with and without cilia in both normal and pathologically altered nasal mucosa.
Two different types of intracytoplasmic lumina have been observed electron microscopically in pseudostratified and metaplastic human nasal mucosa: one type contains microvilli and the other contains both microvilli and cilia. The latter type was seen only in pseudostratified columnar epithelium, while the former was also observed in different stages of metaplasia. From these observations we conclude that the type of intracytoplasmic lumina is determined by the differential potential of the cells in which they occur. Serial sectioning demonstrated that these structures sometimes communicate with the extracellular space. The distinctly different ultrastructure of microvilli lining the intracytoplasmic lumina and the foldlike extensions of the cell membrane favors an intracellular genesis rather than a formation by invagination of the cell membrane. It is suggested that intracytoplasmic lumina occur as a consequence of dysregulation in the process of epithelial maturation.


The patterns of EEG changes in early-onset Parkinson's disease patients.
The aim of this study was to investigate the patterns of brain activity changes in early-onset Parkinson's disease (EOPD) patients and its relationship with the severity of disease and motor deterioration. Electroencephalography (EEG) was performed in a sample of 52 nondemented EOPD patients and 20 healthy controls with similar age. All patients underwent a battery to assess PD severity and motor deterioration. The EEG data were rated by visual and quantitative analyses with the grant total EEG (GTE) score and NicoletOne Software. The parameters of relative band power and coherences for various frequency bands were calculated. In addition, all parameters were compared between groups and examined for correlations with the severity of disease and motor deterioration. The GTE score and two subscores including "Diffuse Slow Waves" and "Frequency of Rhythmic Background Activity" of EOPD increased comparing to control group. The relative beta band powers in seven regions (O1,O2,T5,T6,P3,P4 and C3) indicated significant decreases in EOPD patients and obvious increases in interhemispheric beta coherences were observed in the midtemporal area and frontal area (T3T4 and F3F4). Furthermore, correlation analyses revealed that longer duration was associated with the subscore of "background wave frequency". The beta frequency bands in the right posterior temporal (T6) showed negative relationship with the modified Hoehn-Yahr grading scores. This study is the first to depict the patterns of EEG changes in EOPD patients without dementia and offer a better understanding of the pathophysiological mechanisms underlying and prognostic purposes in EOPD. Some of these changes could serve as useful biomarkers in the study of EOPD.


Surprise, defence, or making sense: what removes hindsight bias?
This paper examines predictions concerning the absence of hindsight bias. Some hypothesise that because hindsight bias increases with outcome "surprisingness", only unsurprising outcomes will remove it. Others suggest the opposite-that very surprising outcomes will reduce or reverse the bias. A proposed sense-making model suggests that unexpected outcomes (i.e., initially surprising) invoke greater sensemaking, which typically produces greater hindsight bias. If the process is not successful, however, the bias may be reduced or reversed. Expected outcomes will also produce little hindsight bias, but only because they invoke relatively little sensemaking in the first place. Feelings of surprise arising from sensemaking (i.e., resultant surprise) should be inversely related to hindsight bias. Results of four experiments provide support for the model. A secondary goal was to determine the boundaries of a defensive-processing mechanism also thought to reduce hindsight bias for negative, self-relevant outcomes. Results suggest that a sense of responsibility for the outcome may be necessary for defensive processing to be activated.


Continuous processing of toxic organics in a fluidized-bed GAC reactor employing carbon replacement.
The fluidized-bed, granular activated carbon (GAC) anaerobic reactor has been shown to be an effective process for the continuous long-term treatment of wastewaters that contain biodegradable or nonbiodegradable toxic organic compounds. With loadings of 10 g COD/kg GAC day, COD removal of 94% was achieved. The anaerobic biofilm that develops on the GAC reduces the load on the carbon by converting the biodegradable organics to methane and carbon dioxide. Approximately 50% of the COD applied to the reactor was converted to methane, thereby reducing carbon requirements. Successful operation of the system requires that a carbon replacement schedule be maintained that will keep the bulk concentrations of toxic adsorbable compounds below their toxic threshold. As long as toxic substances can be adsorbed by the carbon, they will not inhibit the anaerobic biofilm. If nonadsorbable toxic compounds are present, processing must be included to reduce these materials to concentrations below their threshold level.


Intraosseous venous drainage of pretibial varices.
In this study, we present the case of a 74-year-old male patient who presented with a painful pretibial swelling. The patient had a history of varices of the ipsilateral leg for which he had undergone stripping and ligation. At physical examination, pretibial varices and an associated soft tissue swelling were found. Ultrasound revealed pretibial varices, one of which caused a defect on the anterior tibial cortex. Plain radiography showed a small subcortical lucency of the tibial shaft. MRI confirmed the presence of pretibial varices, one of which perforated through the anterior tibial cortex and then coursed intramedullary as an enlarged intraosseous vein. Based on these findings, the diagnosis of varices with an intraosseous venous drainage anomaly was made. The patient was subsequently successfully treated by ambulatory minisurgical phlebectomy. Knowledge and recognition of this intraosseous venous drainage anomaly, which is a rare condition, is pivotal for correct patient management. We review clinical and imaging findings, and discuss previously reported cases.


Co-Occurring Hemolysis and Methemoglobinemia After COVID-19 Infection in Patient With G6PD Deficiency.
Hemolytic anemia and methemoglobinemia are known complications in patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency. They can be elicited by various oxidative stressors. Here we report a case of an adult with the first episode of G6PD deficiency associated hemolysis and methemoglobinemia after acquiring COVID-19 infection, who had no recent exposure to oxidative drugs or fava beans. A 52-year-old gentleman known to have myocardial bridging on aspirin and beta-blocker, with no other medical illnesses, developed anemia symptoms, jaundice, and hypoxia after contracting COVID-19 infection. Further laboratory work revealed non-immune hemolytic anemia, methemoglobinemia, and a positive G6PD screen test. He was treated conservatively with a blood transfusion, and his oxygen saturation improved thereafter. With the widespread COVID-19 infection and its morbidity worldwide, it is crucial to consider methemoglobinemia in the differential diagnosis of hypoxia. Testing for G6PD is an essential next step in such cases, as starting methylene blue in G6PD deficiency can worsen hemolysis. Apart from COVID-19, there is no other identified trigger for the acute event in this patient. It is not known whether COVID-19 infection alone is enough to result in G6PD deficiency-associated hemolysis and methemoglobinemia simultaneously.


F- and G-actin homeostasis regulates mechanosensitive actin nucleation by formins.
Physical force evokes rearrangement of the actin cytoskeleton. Signalling pathways such as tyrosine kinases, stretch-activated Ca(2+) channels and Rho GTPases are involved in force sensing. However, how signals are transduced to actin assembly remains obscure. Here we show mechanosensitive actin polymerization by formins (formin homology proteins). Cells overexpressing mDia1 increased the amount of F-actin on release of cell tension. Fluorescence single-molecule speckle microscopy revealed rapid induction of processive actin assembly by mDia1 on cell cortex deformation. mDia1 lacking the Rho-binding domain and other formins exhibited mechanosensitive actin nucleation, suggesting Rho-independent activation. Mechanosensitive actin nucleation by mDia1 required neither Ca(2+) nor kinase signalling. Overexpressing LIM kinase abrogated the induction of processive mDia1. Furthermore, s-FDAPplus (sequential fluorescence decay after photoactivation) analysis revealed a rapid actin monomer increase on cell cortex deformation. Our direct visualization of the molecular behaviour reveals a mechanosensitive actin filament regeneration mechanism in which G-actin released by actin remodelling plays a pivotal role.


PET positive generalized lymphadenopathy and splenomegaly following interferon-alfa-2b adjuvant therapy for melanoma.
Interferon-alfa (IFN-alpha) is used in patients with various inflammatory and neoplastic disorders. We recently encountered fluorodeoxyglucose (FDG) uptake in generalized lymphadenopathy, splenomegaly, and the marrow in a patient receiving high dose interferon-alpha-2b (IFN-alpha) as adjuvant therapy for the treatment of malignant melanoma. Biopsy of an enlarged hypermetabolic axillary lymph node revealed only a reactive node. Discontinuation of IFN-alpha caused regression of splenomegaly and lymphadenopathy within 3 months.A marrow "hyperstimulation" pattern that can include the spleen is a well-recognized phenomenon on (fluorodeoxyglucose) FDG positron emission tomography/computed tomography (PET/CT) scans in patients receiving chemotherapeutic agents like interferon, though classically due to colony stimulating factors, but does not generally include lymphadenopathy. This case, which likely reflects an interferon-induced pseudolymphoma, highlights the importance of a drug history and clinical correlation for the proper interpretation of FDG PET/CT scans.


Predicting Value of ALCAM as a Target Gene of microRNA-483-5p in Patients with Early Recurrence in Hepatocellular Carcinoma.
The long-term survival rate of hepatocellular carcinoma (HCC) is poor. One of the reasons for the poor rate of survival is the high rate of recurrence caused by intrahepatic metastas is that adversely affects long-term outcome. Many studies have indicated that microRNAs play an important role in HCC, but there has been no research of clonal origins on recurrent HCC (RHCC) by analzing microRNAs. In the present study, we found that miR-483-5p was significantly upregulated in RHCC tissues of short-term recurrence (≤ 2 years) by miRNA microarray screening, and can significantly promote migration and invasion of HCC cells in vitro and increase intrahepatic metastasis in nude mice in vivo. Furthermore, we demonstrated that activated leukocyte cell adhesion molecule (ALCAM), which significantly suppressed migration and invasion of HCC cells, was a direct target of miR-483-5p, and the re-introduction of ALCAM expression could antagonize the promoting effects of miR-483-5p on the capacity of HCC cells for migration and invasion. In addition, expression level of ALCAM was negatively correlated with microvascular invasion and tumor size recognized as prognostic factors. The cases which were negative for ALCAM expression had shorter time to recurrence than positive cases, and univariate and multivariate survival analyses showed that ALCAM was an independent risk factor of HCC recurrence. qRT-PCR and Western blotting showed that the expression of EMT related genes (MMP-2, MMP-9, E-caherin and vimentin) significantly changed as a result of interfering or overexpression of ALCAM, and ALCAM was significantly associated with EMT in HCC. These results suggest that the miR-483-5p/ALCAM axis is an important regulator in invasion and metastasis and biomarker for recurrence risk assessment of HCC.


Cisplatin represses transcriptional activity from the minimal promoter of the O6-methylguanine methyltransferase gene and increases sensitivity of human gallbladder cancer cells to 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-2-chloroethyl)-3-nitrosourea.
O(6)-methylguanine methyltransferase (MGMT) repairs O(6)-alkylguanine in cellular DNA introduced by the clinically used alkylating drug 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU). Thus, cancer cells with MGMT expression are resistant to ACNU treatment. Cisplatin has been reported to suppress MGMT expression; however, the molecular mechanism by which cisplatin reduces MGMT expression remains to be elucidated. Using gallbladder cancer cells (KMG-C) expressing MGMT, we analyzed whether a low dose of cisplatin suppresses MGMT expression, followed by an enhanced drug effect of ACNU in vitro and in vivo. We also investigated the promoter region critical for the transcriptional repression of MGMT gene by cisplatin using 5 deletion mutants in reporter promoter assays. In RT-PCR analysis, the expression of MGMT mRNA in KMG-C cells was dose- and time-dependently repressed. Drug sensitivity to ACNU was increased 2-fold by pretreatment with cisplatin, compared with only ACNU treatment, in MTT assays as well as tumor-bearing nude mice. Although the 5'-flanking region is deleted as far as -69 bp upstream of the transcription start site, cisplatin dose dependently inhibited luciferase activity. However, cisplatin did not cause such repression when 59 bp region from -69 to -10 bp was deleted. We confirmed that cisplatin enhanced sensitivity to ACNU in KMG-C cells expressing MGMT both in vitro and in vivo. Furthermore, a low dose of cisplatin repressed the transcription of the MGMT promoter. The 59 bp region in the MGMT promoter was crucial for repression by cisplatin. These results might form the basis of a chemotherapeutic strategy involving alkylating agents via prior cisplatin-induced biochemical modulation.


[Identification of adenoviruses serotypes implicated in haemorrhagic conjunctivitis in Tunisia].
Human adenoviruses (ADV) are distributed worldwide; they are associated with a variety of diseases. Some ADV can be implicated in large epidemics of conjunctivitis, gastroenteritis and respiratory infections. Classical diagnosis of ADV infections is based on virus isolation on cell culture and identification of the serotype by neutralization test or hemagglutination inhibition assay. However, these methods have a lack of rapidity that makes them impractical in clinical situations. With the advent of PCR, the diagnosis of ADV was improved. In this work, we have used molecular techniques for the identification of ADV serotypes implicated in conjunctivitis in Tunisia. A total of 199 conjunctival swabs received between October 2000 and May 2005 were investigated. Serotype identification was performed using a PCR followed by restriction enzyme analysis in the hexon gene. Typing by sequencing of the PCR product was used to confirm the serotype identification. Among the 199 tested clinical specimens, 24% were positive for ADV. Two different profiles were observed: one predominant corresponding to the majority of the detected ADV; this profile is in favour of two distinct serotypes, ADV37 or ADV8; the second profile was specific of ADV4 and was found in one case observed in 2005. Sequencing confirmed two serotypes: ADV8 with an endemoepidemically circulation in our country and ADV4 that appeared sporadic. The present work showed the importance of molecular techniques not only for ADV detection but also for identification of the circulating serotypes. These techniques are practical and interesting mainly for the rapid virological investigation during epidemics.


A three-component Breakfast Quality Score (BQS) to evaluate the nutrient density of breakfast meals.
Nutrient profiling methods can be applied to individual foods or to composite meals. This article introduces a new method to assess the nutrient density of breakfast meals. This study aimed to develop a new breakfast quality score (BQS), based on the nutrient standards previously published by the International Breakfast Research Initiative (IBRI) consortium. BQS was composed of three sub-scores derived from the weighted arithmetic mean of corresponding nutrient adequacy: an eLIMf sub-score (energy, saturated fat, free sugars, and sodium), a PF (protein and fiber) sub-score, and a VMn1 - 14 micronutrient sub-score, where n varied from 0 to 14. The effects of assigning different weights to the eLIMf, PF, and VMn were explored in four alternative models. The micronutrients were calcium, iron, potassium, magnesium, zinc, vitamin A, thiamin, riboflavin, niacin, vitamin B5, vitamin B6, vitamin B12, vitamin C, and vitamin D. Micronutrient permutations were used to develop alternate VMn1 - 14 sub-scores. The breakfast database used in this study came from all breakfasts declared as consumed by adults (>18 years old) in the French dietary survey INCA3. All models were tested with respect to the Nutrient Rich Food Index (NRF9.3). BQS sensitivity was tested using three prototype French breakfasts, for which improvements were made. The correlations of the models with NRF9.3 improved when the VMn>3 sub-score (n > 3) was included alongside the PF and eLIMf sub-scores. The model with (PF+VMn) and eLIMf each accounting for 50% of the total score showed the highest correlations with NRF9.3 and was the preferred final score (i.e., BQS). BQS was sensitive to the changing quality of three prototype breakfasts defined as tartine, sandwich, and cereal. The proposed BQS was shown to valuably rank the nutritional density of breakfast meals against a set of nutrient recommendations. It includes nutrients to limit along with protein, fiber, and a variable number of micronutrients to encourage. The flexible VMn sub-score allows for the evaluation of breakfast quality even when nutrient composition data are limited.


Necrotising fasciitis in the central part of the body: diagnosis, management and review of the literature.
Necrotising fasciitis (NF) is characterised by rapidly spreading necrosis of the soft tissue and fascia. It is rare but can be fatal when not managed properly. The aim of this study is to evaluate the diagnosis, treatment and results such as mortality, morbidity and reconstructive options of NF localised in the central part of the body. The main goal is to emphasise upon the clinical symptoms for early diagnosis which is the most important factor in saving the lives of these patients. Between January 2000 and December 2010, 30 patients with NF localised in central parts of the body were treated. Six of the patients were female (20%) and the others were male (80%). The mean age was 54·03 years (ranged between 26 and 83 years). The average time from the onset of symptoms to diagnosis was 6 days, ranging from 2 to 11 days. The localisation of NF was perineum in 24 patients (80%); inguinal and thigh region in 5 patients (16·7); and back in 1 patient (3·3%). The hospitalisation time was varying between 17 and 32 days (mean 23 days). Six patients (20%) died and 24 patients (80%) survived. All non-survivors had risk factors and secondary comorbidities such as immunosuppression, chronic cardiac failure, and diabetes with high glucose level. Survivors also underwent repeated debridement operation 2-4 times. Reconstructive procedures were split-thickness skin graft (STSG) in eight patients (33·3%), fasciocutaneous flaps in four patients (16·6%), fasciocutaneous flap + STSG in six patients (25%), scrotal flap + STSG in two patients (6·6%), scrotal flap in two patients (6·6%) and musculocutaneous flap + STSG in one patient (3·3%). There was no major complication such as flap and graft loss, after reconstructive procedures. Early diagnosis of NF may be the lifesaving factor. Amuputation can save the patient's life in the case of NF in the extremities; however, this is not an option for NF in central parts of the body. In these cases, when NF is suspected, early debridement of necrotic tissues should be performed. As soon as the infection and the spread of the necrosis are controlled, reconstruction should be considered.


Germline mutations of DICER1 in Chinese women with BRCA1/BRCA2-negative familial breast cancer.
Germline mutations in identified breast cancer susceptibility genes account for less than 20% of Chinese familial breast cancers. Dicer is an essential component of the microRNA-producing machinery; germline mutations of DICER1 have been confirmed in familial pleuropulmonary blastoma, ovarian sex cord-stromal tumors, and other cancers. Low expression of DICER1 is frequently detected in breast cancer. However, whether germline mutations of DICER1 occur in familial breast cancers remain unknown. Sixty-five breast cancer probands from BRCA1/BRCA2-negative Chinese breast cancer families were screened for germline mutations in DICER1. In addition, 100 unrelated healthy females were enrolled as controls. A polymerase chain reaction sequencing assay was used to screen for mutations in coding regions and at the exon-intron boundaries of DICER1. All variants in introns were evaluated using the NNSplice software to determine the potential splicing effect. A total of 12 germline variants were found, including 11 variants in introns and 1 variant in the 3'-non-coding region. Four variants (IVS8-205 C>T, IVS11+131 delGAAA, IVS16+42 delTA, and IVS19+160 T>C) were novel. Three variants (IVS11+105 C>T, IVS16+42 delTA, and 6095 T>A) may affect splice sites. None of the observed variants appeared to be disease-related, suggesting that germline mutations in DICER1 are rare or absent in familial breast cancer patients.


Rhodococcus equi peritonitis in continuous ambulatory peritoneal dialysis: a first in Australia.
A 33-year-old Caucasian man with end-stage renal disease secondary to biopsy-proven IgA nephropathy, managed with continuous ambulatory peritoneal dialysis (PD), presented with PD-related peritonitis, the causal organism being a non-branching Gram-positive bacillus, Rhodococcus equi. Initial empirical Gram positive and negative coverage with cefazolin and ceftazidime was unsuccessful, but following isolation of the organism, and conversion to intraperitoneal vancomycin and oral ciprofloxacin, the peritonitis episode resolved. At day 10, vancomycin was switched to azithromycin for a total of 6 weeks of antimicrobial therapy. The PD catheter was preserved, and the patient remained peritonitis-free at 6 months of follow-up.


Purification of recombinant human cPLA2 gamma and identification of C-terminal farnesylation, proteolytic processing, and carboxymethylation by MALDI-TOF-TOF analysis.
Cytosolic phospholipase A(2)gamma (cPLA(2)gamma) is a calcium-independent, membrane-associated phospholipase A(2) that possesses a C-terminal prenylation motif (-CCLA) whose covalent structure cannot be deduced from the primary sequence alone. Accordingly, we overexpressed human cPLA(2)gamma containing an N-terminal His tag ((His)(6)cPLA(2)gamma) in Sf9 cells and quantitatively solubilized and purified the enzyme by sequential immobilized metal affinity and Mono Q column chromatographies. The final preparation appeared as a single 61 kDa band after SDS-PAGE/silver-staining, possessed high lysophospholipase activity (50 micromol min(-1) mg(-1)), and was inhibited by, but did not hydrolyze, palmitoyl-CoA. Radiolabeling of recombinant human cPLA(2)gamma with [(3)H]-mevalonolactone in the absence of statins and subsequent cleavage of prenyl groups with Raney nickel revealed that the enzyme is only farnesylated and is not geranylgeranylated. Analysis of CNBr-digested cPLA(2)gamma by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI/TOF-TOF) mass spectrometry demonstrated the presence of a farnesyl moiety at Cys-538, cleavage of the Cys(538)-Cys(539) bond, and carboxymethylation of the resultant C-terminal prenylated cysteine. Collectively, these results describe the solubilization and purification of recombinant cPLA(2)gamma to homogeneity and identify cPLA(2)gamma as a farnesylated protein that undergoes at least three sequential posttranslational modifications that likely facilitate its targeting and interactions with its membrane substrates.


Systematic nomenclature of picornavirus proteins.
An easily learned convention for systematizing the nomenclature of picornavirus proteins is described. The convention is based upon an idealized map, called the L434 diagram, of the picornavirus polyprotein.


CN(x) nanotubes with pyridinelike structures: p-type semiconductors and Li storage materials.
Using density functional theory (DFT) computations, we investigated pyridinelike structures in a series of single-walled CN(x) nanotubes and found that tube chirality and diameter play important roles in the formation of pyridinelike structures. Not pyridinelike structures but more N(C) defects (N atoms substituting for C atoms) should be responsible for the experimentally observed donor states of CN(x) nanotubes. The adsorption energies of Li at the pyridinelike defect are so large and the energy barrier for lithium penetrating the defect is so low that CN(x) nanotubes with pyridinelike structures have enhanced capability for lithium storage.


Economic evaluation of 5-HT3 receptor antagonists in combination with dexamethasone for the prevention of 'overall' nausea and vomiting following highly emetogenic chemotherapy in Chinese adult patients.
Background Two pivotal Phase III trials compared the efficacy of palonosetron, ondansetron and granisetron, combined with dexamethasone, for the prevention of nausea and vomiting following highly emetogenic chemotherapy. However, an economic evaluation of these three regimens in the real-world setting of Chinese adult patients has not been determined. Objectives To estimate, from the perspective of the Chinese healthcare system, which of these frequently used strategies consisting of 0.25 mg palonosetron (0.25P), 16 mg ondansetron (Onda), and 3 mg granisetron (Gran), is the most cost-effective option in patients following highly emetogenic chemotherapy. Methods A Markov decision-analytic model was developed. The health and economic outcomes of the three strategies; 0.25P, Onda, and Gran were investigated. The clinical and utility data were taken from published studies. The cost data were calculated according to current local Chinese practices. Sensitivity analyses were performed to determine the impact of uncertainty regarding the results. Results The base-case analysis showed that the 0.25P strategy yielded maximum health benefits compared with the other two strategies. However, the probabilistic sensitivity analysis demonstrated that the Gran strategy was the most cost-effective approach when the willingness-to-pay threshold was not more than US$22,515/quality-adjusted life year. Moreover, palonosetron is not cost-effective in preventing 'overall' nausea and vomiting following highly emetogenic chemotherapy in Chinese patients. Conclusions Our analysis suggests that, compared with palonosetron and ondansetron, 3 mg granisetron may be a cost-effective treatment option in the current Chinese healthcare setting.


Columnar mesophases of hexabenzocoronene derivatives. II. Charge carrier mobility.
Combining atomistic molecular dynamic simulations, Marcus-Hush theory description of charge transport rates, and master equation description of charge dynamics, we correlate the temperature-driven change of the mesophase structure with the change of charge carrier mobilities in columnar phases of hexabenzocoronene derivatives. The time dependence of fluctuations in transfer integrals shows that static disorder is predominant in determining charge transport characteristics. Both site energies and transfer integrals are distributed because of disorder in the molecular arrangement. It is shown that the contributions to the site energies from polarization and electrostatic effects are of opposite sign for positive charges. We look at three mesophases of hexabenzocoronene: herringbone, discotic, and columnar disordered. All results are compared to time resolved microwave conductivity data and show excellent agreement with no fitting parameters.


Purified Shigella enterotoxin does not alter intestinal motility.
A purified Shigella enterotoxin (pST) and a cell-free lysate with pST removed (CFL-pST) from the whole-cell lysate of Shigella dysenteriae 60 R were used to study their effect on the myoelectric activity and mucosal integrity of rabbit ileal segments. We have previously defined two myoelectric patterns: the migrating action potential complex and repetitive bursts of action potentials that occur in response to certain bacteria and their enterotoxins. The in vivo model consisted of isolated ileal segments in male New Zealand White rabbits. The segments were infused with sterile saline (1 ml/h), pST (2.4-micrograms injection), or CFL-pST (1 ml/h). Myoelectric activity in the segments exposed to pST was similar to that with the saline infusion, but CFL-pST induced significant alterations in myoelectric activity in the form of repetitive bursts of action potentials. The mucosa of the segments exposed to pST showed only mild inflammatory changes. In contrast, CFL-pST caused moderate to severe inflammatory changes with enterocyte necrosis. These studies show that pST, a known enterotoxin, did not alter myoelectric activity and had no significant effect on the integrity of ileal mucosa, as determined by light microscopy. CFL-pST caused both inflammation and tissue necrosis with significant alterations in motor activity. These studies suggest that S. dysenteriae 60 R produces a substance or substances other than pST that cause florid in vivo cytotoxicity and alter myoelectric activity.


Physical and degradation properties of PLGA scaffolds fabricated by salt fusion technique.
Tissue engineering scaffolds require a controlled pore size and interconnected pore structures to support the host tissue growth. In the present study, three dimensional (3D) hybrid scaffolds of poly lactic acid (PLA) and poly glycolic acid (PGA) were fabricated using solvent casting/particulate leaching. In this case, partially fused NaCl particles were used as porogen (200-300µ) to improve the overall porosity (≥90%) and internal texture of scaffolds. Differential scanning calorimeter (DSC) analysis of these porous scaffolds revealed a gradual reduction in glass transition temperature (Tg) (from 48°C to 42.5°C) with increase in hydrophilic PGA content. The potential applications of these scaffolds as implants were further tested for their biocompatibility and biodegradability in four simulated body fluid (SBF) types in vitro. Whereas, simulated body fluid (SBF) Type1 with the optimal amount of HCO3 (-) ions was found to be more appropriate and sensible for testing the bioactivity of scaffolds. Among three combinations of polymer scaffolds, sample B with a ratio of 75:25 of PLA: PGA showed greater stability in body fluids (pH 7.2) with an optimum degradation rate (9% to 12% approx). X-ray diffractogram also confirmed a thin layer of hydroxyapatite deposition over sample B with all SBF types in vitro.


Allergic Bronchopulmonary Aspergillosis.
Allergic bronchopulmonary aspergillosis (ABPA), a progressive fungal allergic lung disease, is a common complication of asthma or cystic fibrosis. Although ABPA has been recognized since the 1950s, recent research has underscored the importance of Th2 immune deviation and granulocyte activation in its pathogenesis. There is also strong evidence of widespread under-diagnosis due to the complexity and lack of standardization of diagnostic criteria. Treatment has long focused on downregulation of the inflammatory response with prolonged courses of oral glucocorticosteroids, but more recently concerns with steroid toxicity and availability of new treatment modalities has led to trials of oral azoles, inhaled amphotericin, pulse intravenous steroids, and subcutaneously-injected anti-IgE monoclonal antibody omalizumab, all of which show evidence of efficacy and reduced toxicity.


Patients with anxiety disorders, stress related disorders and somatoform disorders in the Psychiatric Emergency Out-patient Clinic.
To establish possible differences in gender, age and urgency between patients with anxiety disorders, stress related disorders and somatoform disorders and patients with other psychiatric diagnoses. 2760 consecutive admissions in the Psychiatric Emergency Out-patient Clinic (PEOC) over the period 2004-2005 were included. Data about diagnosis (ICD-10), gender, age and urgency were gathered from medical documentation. Data were compared between two groups of patients according to diagnosis; the first group encompassed anxiety disorders, stress related disorders and somatoform disorders (F 40-F 48) and the second group gathered all other psychiatric diagnoses (others). One third (36%) of patients examined in PEOC suffered for anxiety disorders, stress related disorders and somatoform disorders. The most common diagnoses within this group were stress related disorders (F 43: 41.7%). Gender ratio was in favour of women in both groups. Significantly more women suffered for anxiety, stress-related and somatoform disorders (63.4% vs. 56.9%, p<0.005). Patients with anxiety, stress-related and somatoform disorders were significantly younger compared with others (40.4 years vs. 45.2 years, p<0.001), and they were significantly less urgent cases compared with others (46.7% vs. 65.3.3%, p<0.001). Anxiety disorders, stress related disorders and somatoform disorders are common in PEOC. The higher percentage of women among these patients is in accordance with data from the literature; lower age might be a consequence of earlier onset of the above mentioned disorders.


The role of multislice computed tomography of the costal cartilage in adult age estimation.
To establish population-specific age estimation models in adults from costal cartilage for contemporary Chinese by using three-dimensional volume-rendering technique. Five hundred and twelve individuals (254 females and 258 males) with documented ages between 20 and 85 years were retrospectively included. Their clinical CT examinations (1 mm slice thickness) were used to develop the sex-specific age prediction model. A validation sample comprising 26 female and 24 male individuals was then used to test the predictive accuracy of the established models. Simple linear regression (SLR), multiple linear regression (MLR), gradient boosting regression (GBR), support vector machine (SVM), and decision tree regression (DTR) were utilized to build the age diagnosis models from calibration samples. By comparison, the decision tree regression was the relatively more accurate age prediction model for male, with mean absolute error = 5.31 years, least absolute error = 0.10 years, correct percentage within 5 years = 54%, and the correct percentage within 10 years = 88%. The stepwise multiple linear regression equations was the relatively more accurate one for female, with mean absolute error = 6.72 years, least absolute error = 0.68 years, correct percentage within 5 years = 42%, and correct percentage within 10 years = 77%. Our results indicated that the present established age estimation model can be applied as an additional guidance for age estimation in adults.


Effects of DJ‑1 on apoptosis and mitophagy of glomerular podocytes.
By studying the effects of DJ-1 overexpression and silencing on the morphological structure and mitophagy of glomerular podocytes, the present study aimed to identify the effects of DJ-1 on glomerular podocyte apoptosis and mitophagy. MPC5 mouse glomerular podocytes were cultured in vitro and divided into four groups: Control, DJ-1 overexpression, empty vector and DJ-1 silencing. DJ-1 gene overexpression and silencing models were prepared, the morphological structures of podocytes and mitochondria in each group were observed, and podocyte apoptosis and DJ-1/PTEN expression were subsequently detected in each group. The experimental results showed reduced volume, retracted foot processes, loosened intercellular connections, presence of dead cells, increased apoptotic rate, increased expression of PTEN, and swollen mitochondria due to the number of vacuoles and autophagosomes in podocytes in the DJ-1 silencing group. The surface areas of podocytes in the DJ-1 overexpression group were greater than those in the control group. Moreover, the structure of the foot processes was more obvious, the number of cells was greater, the intercellular connections were closer, the apoptotic rate was reduced, the expression of PTEN was decreased, the mitochondrial structure was more obvious and the mitochondrial cristae were more whole. Notably, there were no differences between the empty vector and control groups. In conclusion, these results indicated that DJ-1 may regulate podocyte apoptosis and mitophagy through the DJ-1/PTEN pathway, and could maintain the stability of the normal morphology, structure and function of glomerular podocytes.


Lymphocyte transformation test for diagnosis of isothiazolinone allergy in man.
The lymphocyte transformation test (LTT) has been used for evaluation of in vitro lymphocyte responses in 18 patients with dermatitis and positive patch tests to 200 ppm of a combination of 5-chloro-2-methylisothiazolinone and 28methylisothiazolinone (MCI) in nine patients with dermatitis unrelated to MCI and in seven subjects without skin diseases. Two workers sensitized by occupational exposure to a formulation containing 1,2-benzisothiazolin-3-one (BIT) were also studied. Lymphocytes from nine patch-test-positive patients proliferated vigorously to MCI in vitro. Lymphocytes from the remaining nine patients were not stimulated. Lymphocytes from two BIT-sensitized workers responded to BIT in vitro. The lymphocyte proliferation to isothiazolinones indicates the presence of memory cells in the patients' blood and confirms immunologic reaction to the inducing agent. To establish clinical relevance of LTT results, 12 MCI patch-test-positive patients underwent "use test" with lotion containing 15 ppm MCI. Four of five LTT-positive patients were use-test-positive, whereas seven of seven LTT-negative patients were use-test-negative. LTT-positive and lotion-positive patients responded to 100 ppm or lower concentrations of MCI on patch testing, whereas seven of eight LTT-negative and lotion-negative patients responded to 200 ppm only. In the case of MCI, proliferation was due to the chlorinated component, indicating that this part contains an allergenic epitope. Finally, MCI-specific lymphocyte proliferation was observed only in patients with MCI-positive skin test, but not in nine patients with dermatitis induced by other agents, or in seven subjects without skin diseases. Thus, the lymphocyte transformation test is able to distinguish between irritant and allergic skin responses. It may also be valuable in establishing the clinically relevant patch-test concentration of allergens with irritative properties.


Drug-induced thrombocytopenia during G-CSF therapy in a patient with chronic neutropenia.
A one-year-old infant presented with recurrent perianal abscesses and pneumonia, and was diagnosed with chronic neutropenia. Treatment with granulocyte colony-stimulating factor (G-CSF) was initiated, and thrombocytopenia was detected three weeks later. The drug was discontinued but the patient's platelet count did not improve. A short course of high-dose methylprednisolone was administered, and both the platelet and neutrophil counts returned to normal. Based on this response, in order to maintain the neutrophil count the steroid treatment was continued for one year with tapered doses. At the time of writing, the patient had been doing well for 13 months without treatment. This report highlights that G-CSF treatment for severe chronic neutropenia in a child may be associated with thrombocytopenia.


One-Pot Construction of 1-Phenylchromeno[3,4- b]pyrrol-4(3 H)-one: Application to Total Synthesis of Ningalin B and a Pyrrolocoumarin-Based Electrochromic Switch.
An efficient construction of 1-phenylchromeno[3,4- b]pyrrol-4(3 H)-one via coupling of 1-styrylpyrrolidine and 4-chloro-3-nitrocoumarin as a key step is reported. This reaction is further applied to the total synthesis of the natural product ningalin B in five linear steps with an overall yield of 41.5% and a pyrrolocoumarin-based electrochromic switch.


Low adherence to recommended pre-participation cardiac evaluation of Swedish athletes Pre-participation cardiac evaluation of athletes is recommended by international organizations like the European Society of Cardiology and the American Heart Association, as well as by the Swedish Sports Confederation. The purpose of the evaluation is to prevent sudden cardiac death in athletes by early identification of individuals at risk. To our knowledge, no previous study has been made regarding the implementation of pre-participation cardiac evaluation of athletes in Sweden. We performed an electronical survey addressing sports clubs in one out of 21 districts in which the Swedish Sports Confederation is geographically divided. Only four out of 22 responding clubs with elite athletes preformed cardiac evaluation. Lack of knowledge about the recommendations as well as how to perform the evaluation were mentioned as reasons not to evaluate the athletes. Our results indicate the need for more information about pre-participation cardiac evaluation of athletes in Sweden.


Cancer stem cells and response to therapy.
The cancer stem cell (CSC) model states that cancers are organized in cellular hierarchies, which explains the functional heterogeneity often seen in tumors. Like normal tissue stem cells, CSCs are capable of self-renewal, either by symmetric or asymmetric cell division, and have the exclusive ability to reproduce malignant tumors indefinitely. Current systemic cancer therapies frequently fail to eliminate advanced tumors, which may be due to their inability to effectively target CSC populations. It has been shown that embryonic pathways such as Wnt, Hedgehog, and Notch control self-renewal and cell fate decisions of stem cells and progenitor cells. These are evolutionary conserved pathways, involved in CSC maintenance. Targeting these p.


Cyclamates: a review of the current position.
Cyclamates were prohibited for use as food additives in the U.S.A. and other parts of the world during 1970 because of the fears of carcinogenicity. The author reviews the evidence leading to this ban and discusses the appeal made against the decision which led to the lifting of the cyclamate restrictions in Australia in November 1974.


Adaptation of the Corticomuscular and Biomechanical Systems of Pianists.
Independent control of movements between the fingers plays a role in hand dexterity characterizing skilled individuals. However, it remains unknown whether and in what manner neuromuscular and biomechanical constraints on the movement independence of the fingers depend on motor expertise. Here, we compared motor dexterity, corticospinal excitability of multiple muscles, muscular activation, and anatomical features of the fingers between the pianists and nonpianists. When the ring finger was passively moved by a robot, passive motions produced at the adjacent fingers were smaller for the pianists than the nonpianists, indicating reduced biomechanical constraint of fingers in the pianists. In contrast, when the ring finger moved actively, we found no group difference in passive motions produced at the adjacent fingers; however, reduced inhibition of corticospinal excitability of the adjacent fingers in the pianists compared with the nonpianists. This suggests strengthened neuromuscular coupling between the fingers of the pianists, enhancing the production of coordinated finger movements. These group differences were not evident during the index and little finger movements. Together, pianists show expertise-dependent biomechanical and neurophysiological adaptations, specifically at the finger with innately low movement independence. Such contrasting adaptations of pianists may subserve dexterous control of both the individuated and coordinated finger movements.


The phenomenon of "picky eater": a behavioral marker in eating patterns of toddlers.
To determine if toddlers who were considered "picky eaters" had lower dietary scores than non-picky eaters, and if family environment and socioeconomic status were significantly related to picky eater status and dietary scores. An incomplete block design provided two interviews at randomly assigned times (24, 28, 32, or 36 months) of Caucasian mothers from upper socioeconomic (n=74) and lower socioeconomic status (n=44). Using trained interviewers, 6 days of food intake, two administrations of a questionnaire about toddler's eating behavior, and one administration of the Family Environment Scales were collected in the home. MANOVA, discriminant function analysis, and logistic regression procedures were used to determine significant differences between picky and non-picky eater groups. Picky eaters had lower dietary variety (p=.03) and diversity scores (p=.009) than non-picky eaters. Mothers of picky eaters compared to those of non-picky eaters used persuasion (p=.0001) and ranked their child's eating behaviors as more problematic (p=.0001). Toddlers perceived by their mothers as picky eaters had significantly lower dietary variety and diversity scores. Parents need information and strategies to increase the number of foods acceptable to their toddlers and to develop a sound feeding plan.


Liver biopsy in a district general hospital: changes over two decades.
To study liver biopsy practice over two decades in a district general hospital in the United Kingdom. We identified all patients who had at least one liver biopsy between 1986 and 2006 from the databases of the radiology and gastroenterology departments. Subjects with incomplete clinical data were excluded from the study. A total of 103 liver biopsies were performed. Clinical data was available for 88 patients, with 95 biopsies. Between 1986 and 1996, 18 (95%) out of the 19 liver biopsies performed were blind and 6 (33%) were for primary biliary cirrhosis. Between 1996 and 2006, 14 (18%) out of 76 biopsies were blind; and the indications were abnormal liver tests (33%), hepatitis C (12%) and targeted-biopsies (11%). Liver biopsies were unhelpful in 5 (5%) subjects. Pain was the most common complication of liver biopsy (5%). No biopsy-related mortality was reported. There was a trend towards more technical failures and complications with the blind biopsy technique. Liver biopsies performed in small district hospitals are safe and useful for diagnostic and staging purposes. Abnormal liver tests, non-alcoholic fatty liver disease and targeted biopsies are increasingly common indications. Ultrasound-guided liver biopsies are now the preferred method and are associated with fewer complications.


[Osteochondrosis dissecans of the knee and sports].
Osteochondritis dissecans (O.d.) of the knee is a common disease, but the aetiological factors are still controversial. With a follow-up examination of 97 surgically treated patients (109 lesions) we looked for the influence of preoperative sports activities as a possible aetiological factor. Most of the patients complained first symptoms at puberty age between 10 and 15 years, girls with a mean age of 12.4 years and boys with a mean age of 15 years. Because of that, humoral factors are supposed to have influence on the onset of this lesion. Also patients with O.d. of the knee demonstrated a high rate of sports activities and/or trauma prior to the onset of symptoms. There was also a good correlation between lesions at the medial condyle with varus malalignment. Additionally, in most of the patients a high grade of general ligament laxity could be observed at the follow-up examination. It seems that biomechanical factors have an important aetiological influence on the initiation of O.d. at the knee. At the date of follow-up examination all patients reported on a minor sports activity in comparison to sports activities prior to the onset of symptoms. The amount of postoperative sports activities depends on the level of the resulting Lysholm score and on the grade of osteoarthritis.


Alterations in aortic and tail artery reactivity to agonists after streptozotocin treatment.
In the present study, responses to various agonists in thoracic aorta and tail artery strips, obtained from 4-week streptozotocin (STZ)-treated rats and age-matched controls, were studied. Responses in aorta obtained from diabetic animals to the alpha-agonists, norepinephrine (NE), and methoxamine (MOX), to calcium (Ca2+) and potassium (K+) were found to be depressed relative to control tissue. Responses in tail artery however, were found to be different. While responses to K+ were decreased and to Ca2+ unchanged, tail artery strips obtained from diabetic animals were found to be supersensitive to both alpha-agonists relative to control tissue. Another observed difference between the two tissues was in their catecholamine content. While induction of diabetes did not alter catecholamine levels in the aorta, a significant decrease was produced in the tail artery, relative to control tissue levels. Our results indicate that inherent differences (such as the degree of innervation) may contribute to the differential responses observed in the two tissues studied. They further suggest the possible involvement of alterations in calcium utilization in aorta and the development of postjunctional supersensitivity in tail artery obtained from STZ-treated animals.


Mechanisms of Inactivation of Dry Escherichia coli by High-Pressure Carbon Dioxide.
High-pressure carbon dioxide processing is a promising technology for nonthermal food preservation. However, few studies have determined the lethality of high-pressure CO2 on dry bacterial cells, and the mechanism of inactivation remains unknown. This study explored the mechanisms of inactivation by using Escherichia coli AW1.7 and mutant strains differing in heat and acid resistance, in membrane composition based on disruption of the locus of heat resistance, and in genes coding for glutamate decarboxylases and cyclopropane fatty acid synthase. The levels of lethality of treatments with liquid, gaseous, and supercritical CO2 were compared. The cell counts of E. coli AW1.7 and mutants with a water activity (aW) of 1.0 were reduced by more than 3 log10 (CFU/ml) after supercritical CO2 treatment at 35°C for 15 min; increasing the pressure generally enhanced inactivation, except for E. coli AW1.7 ΔgadABE. coli AW1.7 Δcfa was more susceptible than E. coli AW1.7 after treatment at 10 and 40 MPa; other mutations did not affect survival. Dry cells of E. coli were resistant to treatments with supercritical and liquid CO2 at any temperature. Treatments with gaseous CO2 at 65°C were more bactericidal than those with supercritical CO2 or treatments at 65°C only. Remarkably, E. coli AW1.7 was more susceptible than E. coli AW1.7 Δcfa when subjected to the gaseous CO2 treatment. This study identified CO2-induced membrane fluidization and permeabilization as causes of supercritical mediated microbial inactivation, and diffusivity was a dominant factor for gaseous CO2IMPORTANCE The safety of dry foods is of increasing concern for public health. Desiccated microorganisms, including pathogens, remain viable over long periods of storage and generally tolerate environmental insults that are lethal to the same organisms at high water activity. This study explored the use of high-pressure carbon dioxide to determine its lethality for dried Escherichia coli and to provide insight into the mechanisms of inactivation. The lethality of high-pressure CO2 and the mechanisms of CO2-mediated inactivation of dry E. coli depended on the physical state of CO2 Liquid and supercritical CO2 were ineffective in reducing the cell counts of dry E. coli isolates, and the effectiveness of gaseous CO2 was related to the diffusivity of CO2 Results provide a novel and alternative method for the food industry to enhance the safety of low aW products.


Three-dimensional geometry of the Heineke-Mikulicz strictureplasty.
The objective of this study was to assess the regional geometry of the Heineke-Mikulicz (HM) strictureplasty. The HM intestinal strictureplasty is commonly performed for the treatment of stricturing Crohn's disease of the small intestine. This procedure shifts relatively normal proximal and distal tissue to the point of narrowing and thus increases the luminal diameter. The overall effect on the regional geometry of the HM strictureplasty, however, has not been previously described in detail. HM strictureplasties were created in latex tubing and cast with an epoxy resin. The resultant casts of the lumens were then imaged using computed tomography. Using 3-dimensional vascular reconstruction software, the cross-sectional areas were determined and the surface geometry was examined. The HM strictureplasty, while increasing the lumen at the point of the stricture, also results in a counterproductive luminal narrowing proximal and distal to the strictureplasty. Within the model used, cross-sectional area was diminished 25% to 50% below baseline. This effect is enhanced when 2 strictureplasties are placed in close proximity to each other. The HM strictureplasty results in alterations in the regional geometry that may result in a compromise of the lumen proximal and distal to the location of the strictureplasty. When 2 HM strictureplasties are created in close proximity to each other, care should be undertaken to assure that the lumen of the intervening segment is adequate.


The life of a skin cancer.
The mechanisms of carcinogenesis and metastasis are becoming better elucidated. The recent literature pertaining to the current understanding of the mechanisms of carcinogenesis and metastasis was reviewed, and some of the basic information that is known about these processes and how they relate to the development of cutaneous cancers is presented in this article. The development of a skin cancer consists of a three-step process of carcinogenesis-initiation, promotion, and progression. When metastasis occurs, a six-step process of complex cellular adaptations enables the tumor cell to metastasize and proliferate successfully. The mechanisms of carcinogenesis and metastasis are pertinent not only to dermatologists and physicians who treat skin cancer but also to any physician who must manage human malignant tumors. Therefore, a general background of information on this subject is an important aspect of medical knowledge for most physicians.


CSF and plasma amyloid-β temporal profiles and relationships with neurological status and mortality after severe traumatic brain injury.
The role of amyloid-β (Aβ) neuropathology and its significant changes in biofluids after traumatic brain injury (TBI) is still debated. We used ultrasensitive digital ELISA approach to assess amyloid-β1-42 (Aβ42) concentrations and time-course in cerebrospinal fluid (CSF) and in plasma of patients with severe TBI and investigated their relationship to injury characteristics, neurological status and clinical outcome. We found decreased CSF Aβ42 levels in TBI patients acutely after injury with lower levels in patients who died 6 months post-injury than in survivors. Conversely, plasma Aβ42 levels were significantly increased in TBI with lower levels in patients who survived. A trend analysis showed that both CSF and plasma Aβ42 levels strongly correlated with mortality. A positive correlation between changes in CSF Aβ42 concentrations and neurological status as assessed by Glasgow Coma Scale (GCS) was identified. Our results suggest that determination of Aβ42 may be valuable to obtain prognostic information in patients with severe TBI as well as in monitoring the response of the brain to injury.


The Ideal Thigh: A Crowdsourcing-Based Assessment of Ideal Thigh Aesthetic and Implications for Gluteal Fat Grafting.
As the popularity of aesthetic gluteoplasty continues to grow, there is renewed focus on defining the ideal buttocks. However, the literature lacks studies characterizing an ideal thigh, despite the impact of thigh contour on overall gluteal aesthetic. The authors performed the first population analysis of the characteristics of perception of attractive thighs, to identify a role for fat grafting of the thigh in gluteoplasty. Survey images were digitally modified to create thighs of varying widths and angles relative to fixed buttocks. Thigh-to-buttock ratios and the buttock-thigh junction were studied. Data were stratified and analyzed according to age, gender, and ethnicity of the respondents. Amazon Mechanical Turk was used as a novel crowdsourcing platform for surveying aesthetic preferences. A total of 1034 responses were included of whom 54.4% were male, and 45.6% were female. All age groups and ethnicities were represented. Overall, 43.8% of respondents preferred the widest buttock-thick junction angle on posterior view. There was no clear preference between larger or smaller thigh-to-hip ratios on lateral view. Characteristics of the ideal thigh include wider thighs with greater horizontal projection, creating a more natural contour from the augmented buttock. These findings represent a paradigm shift from the traditionally assumed preference for slender thighs. Plastic surgeons should carefully consider thigh anatomy in their gluteal augmentation patients, as simultaneous thigh augmentation may lead to a more aesthetically pleasing outcome. Further research is needed into best practices and techniques to attain ideal thigh proportions.


Apical and root canal space sealing abilities of resin and glass ionomer-based root canal obturation systems.
To investigate the apical sealing ability of glass ionomer and resin-based root canal obturation systems in comparison to a conventional vertical compaction of warm guttapercha. Forty-five extracted human teeth were randomly assigned into 3 groups of 15 each: a resin-based (EndoRez), a glass ionomer-based (Activ GP), and a conventional gutta-percha plus pulp sealer obturation system (GP/EWT). Apical and root canal space sealing abilities were assessed on five cross-sections 1.0 mm apart starting from the apex. Cross-section images were analysed using a focus-variation 3D scanning microscope and unsealed space was calculated as the percentage of total root canal space occupied by voids and debris. EndoRez had significantly higher rate of apical leakage and deeper dye penetration as compared to GP/EWT and Activ GP. EndoRez group had also more voids and debris (22.5%) in the root canal spaces as compared to GP/EWT (10.5%) and Activ GP (10.8%). Apical leakages occurred not only along the root canal walls, but also along the gutta-percha cones with EndoRez as a result of significant polymerisation shrinkage of the resin sealer. Resin-based EndoRez did not form an adequate apical seal of filled root canals. Glass ionomer-based Activ GP was comparable to a vertical compaction of warm guttapercha plus EWT sealer in sealing root canal spaces.


Grasping two-dimensional images and three-dimensional objects in visual-form agnosia.
Visually guided prehension is controlled by a specialized visuomotor system in the posterior parietal cortex. It is not clear how this system responds to visual stimuli that lack three-dimensional (3D) structure, such as two-dimensional (2D) images of objects. We asked a neurological patient with visual-form agnosia (patient D.F.) to grasp 3D objects and 2D images of the same objects and to estimate their sizes manually. D.F.'s grip aperture was scaled to the sizes of the 2D and 3D target stimuli, but her manual estimates were poorly correlated with object size. Control participants demonstrated appropriate size-scaling in both the grasping and manual size-estimation tasks, but tended to use a smaller peak aperture when reaching to grasp 2D images. We conclude that: (1) the dorsal stream grasping system does not discriminate in a fundamental way between 2D and 3D objects, and (2) neurologically normal participants might adopt a different visuomotor strategy for target objects that are recognized to be ungraspable. These findings are consistent with the view that the dorsal grasping system accesses a pragmatic, spatial representation of the target object, whereas the ventral system accesses a more comprehensive, volumetric description of the object.


Cutaneous plasmacytic infiltrates.
This article reviews a group of skin disorders that have in common the presence of a cellular infiltrate with plasma cells upon histopathologic examination. These include inflammatory and neoplastic diseases and a group of miscellaneous disorders of varied etiology. The correlation between the presence of plasma cells in the cellular infiltrate and the diagnosis, prognosis, and pathogenesis of the disorders is emphasized.


Tornado-associated fatalities--Arkansas, 1997.
On March 1, 1997, approximately nine tornadoes originating from two separate thunderstorms swept across Arkansas, from Hempstead County in the southwest to Clay County in the northeast (approximately 260 miles). The tornadoes caused 26 deaths and an estimated $115 million in property damage, reflecting damage to residences, nonresidential buildings, bridges, and roads and agriculture and timber losses. The strongest tornadoes touched down southwest of Little Rock in Clark, Saline, and Pulaski counties; the estimated widths of the tornado paths ranged from 1/2 to 1 mile, and wind speeds were > 200 miles per hour (National Weather Service [NWS], unpublished data, 1997). This report summarizes circumstances of the tornado-associated fatalities from information collected by the American Red Cross (ARC); 14 of the 26 fatalities occurred among persons who were in mobile homes.


3'-azido-3'-deoxy-5'-O-isonicotinoylthymidine, a new prodrug of zidovudine. Synthesis, solid state characterization, and anti HIV-1 activity.
Synthesis, solid state characterization and anti HIV-1 activity of 3'-azido-3'-deoxy-5'-O-isonicotinoylthymidine (2), a new prodrug of zidovudine (AZT, 1), are described. Two solid forms of 2 prepared by crystallization from ethyl acetate-petroleum ether (form alpha) and from a melt sample of form alpha (amorphous form) were characterized by X-ray diffractometry, infrared spectroscopy, differential scanning calorimetry (DSC) and thermogravimetry (TGA) techniques. The novel nucleoside exhibited antiviral activity against standard and resistant strain panels of HIV-1 as well as cytotoxicity similar to that of AZT.


Immunoglobulin class-specific antibody response in respiratory syncytial virus infection measured by enzyme immunoassay.
Immunoglobulin class-specific enzyme immunoassay (EIA) was used for determination of antibody responses in sera collected from 26 children with acute primary respiratory syncytial virus (RSV) infections. All 26 patients had IgG antibody responses with a significant titer increase in 24 (92%); an IgM antibody response was detected in 19 of the 26 (73%). From patients aged 6 months or less only 5 of 8 produced detectable IgM antibodies, whereas all patients aged 1-2 years did so. IgM antibodies appeared within 1 week after onset of illness and persisted from 20 days to 2-3 months. An IgA antibody response was observed in 20 of 26 (77%) patients with a significant titer increase detected in 17 of 26 (65%) patients. In some patients the persistence of IgA antibodies followed that of the IgM antibodies, but in others the IgA antibody titers remained stable up to the end of the follow-up. The most sensitive assay system for serological diagnosis of acute RSV infection in children was the determination of titer increases by IgG antibody.


Commercial considerations in tissue engineering.
Tissue engineering is a field with immense promise. Using the example of an early tissue-engineered skin implant, Dermagraft, factors involved in the successful commercial development of devices of this type are explored. Tissue engineering has to strike a balance between tissue culture, which is a resource-intensive activity, and business considerations that are concerned with minimizing cost and maximizing customer convenience. Bioreactor design takes place in a highly regulated environment, so factors to be incorporated into the concept include not only tissue culture considerations but also matters related to asepsis, scaleup, automation and ease of use by the final customer. Dermagraft is an allogeneic tissue. Stasis preservation, in this case cryopreservation, is essential in allogeneic tissue engineering, allowing sterility testing, inventory control and, in the case of Dermagraft, a cellular stress that may be important for hormesis following implantation. Although the use of allogeneic cells provides advantages in manufacturing under suitable conditions, it raises the spectre of immunological rejection. Such rejection has not been experienced with Dermagraft. Possible reasons for this and the vision of further application of allogeneic tissues are important considerations in future tissue-engineered cellular devices. This review illustrates approaches that indicate some of the criteria that may provide a basis for further developments. Marketing is a further requirement for success, which entails understanding of the mechanism of action of the procedure, and is illustrated for Dermagraft. The success of a tissue-engineered product is dependent on many interacting operations, some discussed here, each of which must be performed simultaneously and well.


Delayed resolution of negative pressure pulmonary edema associated with low-molecular-weight dextran after microsurgery--a case report.
Negative pressure pulmonary edema (NPPE) is an uncommon perioperative complication after total or partial upper airway obstruction during anesthesia or respiratory distress. The cardinal event causative of the pathophysiology is believed to be the generation of a sudden and marked negative intrapleural pressure against a closed glottis, resulting in a disruption of the normal intravascular Starling mechanism. Ultimately, excessive intravascular fluid which transudes into the interstitial space eventuates in pulmonary edema. In most reports, patients suffering from NPPE could recover within 24 h after appropriate treatment. Here, we report a case of highly suspected NPPE with delayed resolution associated with the perioperative use of dextran, following microsurgery for digital anastomosis of the nearly amputated right thumb. We have reviewed the possible pathomechanisms of NPPE, and discuss herein the possible relationship between the delayed resolution of NPPE and perioperative use of dextran.


The fatty acid status of rural school children with helminth infections.
Intestinal helminth infections may reduce nutrient availability and thereby impair cognitive growth and other functions. Certain nutrients such as fatty acids may also modulate inflammation and the immune response and, thus, the severity of helminth infection. Fatty acid status may, therefore, both be affected by and affect helminth infection. Rural school-children in South Africa with a 35% helminth infestation rate were assessed for serum total fatty acid status. With hookworm, a higher percentage total n-3 fatty acids were found with functional implications.


Prognostic Relevance of the Lung Ultrasound Score: A Multioutcome Study in Infants with Respiratory Distress Syndrome.
There is growing evidence for the usefulness of the lung ultrasound score (LUS) in neonatal intensive care. We evaluated whether the LUS is predictive of outcomes in infants with respiratory distress syndrome (RDS). Neonates less than 34 weeks of gestational age were eligible for this prospective, multicenter cohort study. The outcomes of interest were the need for mechanical ventilation (MV) at <72 hours of life, the need for surfactant (SF), successful weaning from continuous positive airway pressure (CPAP), extubation readiness, and bronchopulmonary dysplasia. Lung scans were taken at 0 to 6 hours of life (Day 1), on Days 2, 3, and 7, and before CPAP withdrawal or extubation. Sonograms were scored (range 0-16) by a blinded expert sonographer. The area under the receiver operating characteristic curve (AUC) was used to estimate the prediction accuracy of the LUS. A total of 647 scans were obtained from 155 newborns with a median gestational age of 32 weeks. On Day 1, a cutoff LUS of 6 had a sensitivity (Se) of 88% and a specificity (Sp) of 79% to predict the need for SF (AUC = 0.86), while a cutoff LUS of 7 predicted the need for MV at <72 hours of life (Se = 89%, Sp = 65%, AUC = 0.80). LUS acquired prior to weaning off CPAP was an excellent predictor of successful CPAP withdrawal, with a cutoff level of 1 (Se = 67%, Sp = 100%, AUC = 0.86). The LUS has significant predictive ability for important outcomes in neonatal RDS. · Lung ultrasound has significant prognostic abilities in neonatal RDS.. · Early sonograms (0-6 h of life) accurately predict the requirement for SF and ventilation.. · Weaning off CPAP is effective when the LUS (range 0-16) is less than or equal to 1..


[Lithium valproate: its neuropharmacological spectrum and effect on transmitter amino acid content of the brain of rats].
The authors studied lithium valproate synthesized at the Institute of Pharmacology, Academy of Medical Sciences of the USSR, and compared it with sodium valproate (depakin, France). It was discovered that like sodium valproate, lithium valproate has a marked anticonvulsant activity, being also effective in experimental seizures induced by bicucullin, exerts a moderate sedative action (potentiation of sodium thiopental and hexenal anesthesia), and a distinct tranquilizing effect in experimental outward inhibition of the conditioned active avoidance response. At the same time the presence in the molecule of lithium ions led to a change in the neurochemical profile of valproate, manifested by an increase in the rat brain content of GABA and glutamate.


Chemical preparation of an isotopically enriched superoxide dismutase and its characterization as a standard for species-specific isotope dilution analysis.
The development of methods to analyze accurately and precisely individual metalloproteins is of increasing importance. Here we describe for the first time the chemical preparation and characterization of an isotopically enriched metalloenzyme containing two different metal isotopes. Its evaluation as a standard in species-specific isotope dilution analysis by HPLC coupled to inductively coupled plasma mass spectrometry is carefully evaluated. Our model enzyme bovine superoxide dismutase (SOD) contains both Cu and Zn and is remarkably stable at high temperatures and even under denaturing conditions. The enzyme's metal cofactors were removed under a range of different conditions and replaced with isotopically enriched 65Cu and 68Zn. Depending on the conditions, various isotopic ratios differing from the natural Cu and Zn abundances were obtained for the reconstituted enzyme. Both the wild type and isotopically enriched enzyme had the same migration pattern on native 1D-PAGE. Using an enzyme activity test, we showed that the incorporated 65Cu was bound to the right SOD-binding site, since the measured activity correlated directly with the amount of Cu incorporated. Mixing the native and the isotopically enriched enzyme standard with free enriched 65Cu and 68Zn or a metal chelator did not result in any exchange or loss of the metals from the enzyme at neutral pH. This verifies the stability of the enzyme metal center under the chosen conditions. The isotopically enriched enzyme standard was spiked into a wild type SOD solution to evaluate its use for species-specific isotope dilution experiments. To our knowledge, this is the first report of the chemical preparation of a metalloenzyme containing two different isotopically enriched metals. We provide evidence that the incorporated isotopically enriched metals are bound to the right binding site of SOD using an specific enzymatic activity assay.


Selective transfer semihydrogenation of alkynes with nanoporous gold catalysts.
A facile, highly chemo- and stereoselective transfer semihydrogenation of alkynes to Z-olefins has been achieved by use of unsupported nanoporous gold (AuNPore) as a heterogeneous catalyst together with formic acid as a hydrogen donor. A variety of terminal/internal and aromatic/aliphatic alkynes were reduced to the corresponding alkenes in high chemical yields with good functional-group tolerance. The catalyst is robust enough to be reused without leaching.


Load-displacement properties of lower cervical spine motion segments.
The load-displacement behavior of 35 fresh adult cervical spine motion segments was measured in compression, shear, flexion, extension, lateral bending and axial torsion tests. Motion segments were tested both intact and with posterior elements removed. Applied forces ranged to 73.6 N in compression and to 39 N in shear, while applied moments ranged to 2.16 Nm. For each mode of loading, principal and coupled motions were measured and stiffnesses were calculated. The effect of disc degeneration on motion segment stiffnesses and the moments required for motion segment failure were also measured. In compression, the stiffnesses of the cervical motion segments were similar to those of thoracic and lumbar motion segments. In other modes of loading, cervical stiffnesses were considerably smaller than thoracic or lumbar stiffnesses. Removal of the posterior elements decreased cervical motion segment stiffnesses by as much as 50%. Degenerated cervical discs were less stiff in compression and stiffer in shear than less degenerated discs, but in bending or axial torsion, no statistically significant differences were evident. Bending moments causing failure were an order of magnitude lower than those for lumbar segments.


Fecal calprotectin in diagnosis and clinical assessment of inflammatory bowel disease.
The fecal neutrophil-derived biomarker calprotectin has several features of an ideal noninvasive test for detecting intestinal inflammation: it is simple, reliable, and low in cost. Its utility in differentiating inflammatory bowel diseases (IBDs) from functional conditions such as irritable bowel syndrome is well documented. Fecal calprotectin (FC) correlates closely with endoscopic activity of IBD. Emerging evidence suggest its usefulness in serial monitoring of disease activity and of therapy success in IBD. A low FC concentration predicts persistence of clinical remission especially in non-symptomatic ulcerative colitis and Crohn's colitis. Here, an overview is given to the current role of FC in diagnosis and clinical assessment of IBD.


Function, disability, and psychological well-being.
Disability research in arthritis, as in disability research in general, has focused on functional limitations and activities of daily living/instrumental activities of daily living (ADL/IADL) disability, and has thus ignored a great deal of daily life. Unfortunately, the areas of life that have been ignored may be those that are most important to individuals, and may also be the most sensitive to the first signs of developing disability. The ability to perform valued life activities, the wide range of activities that individuals find meaningful or pleasurable above and beyond activities that are necessary for survival or self-sufficiency, has strong links to psychological well-being--in some cases, stronger links than functional limitations and disability in basic activities of daily living. A broader assessment of disability has great potential for interrupting the disablement and distress process, thereby improving the quality of life of individuals with arthritis. Assessment of the effects of arthritis, pain, or other chronic health conditions should expand beyond assessment of functional limitations and disability in basic activities to include assessment of disability in advanced, valued activities.


pKa modulation of the acid/base catalyst within GH32 and GH68: a role in substrate/inhibitor specificity?
Glycoside hydrolases of families 32 (GH32) and 68 (GH68) belong to clan GH-J, containing hydrolytic enzymes (sucrose/fructans as donor substrates) and fructosyltransferases (sucrose/fructans as donor and acceptor substrates). In GH32 members, some of the sugar substrates can also function as inhibitors, this regulatory aspect further adding to the complexity in enzyme functionalities within this family. Although 3D structural information becomes increasingly available within this clan and huge progress has been made on structure-function relationships, it is not clear why some sugars bind as inhibitors without being catalyzed. Conserved aspartate and glutamate residues are well known to act as nucleophile and acid/bases within this clan. Based on the available 3D structures of enzymes and enzyme-ligand complexes as well as docking simulations, we calculated the pKa of the acid-base before and after substrate binding. The obtained results strongly suggest that most GH-J members show an acid-base catalyst that is not sufficiently protonated before ligand entrance, while the acid-base can be fully protonated when a substrate, but not an inhibitor, enters the catalytic pocket. This provides a new mechanistic insight aiming at understanding the complex substrate and inhibitor specificities observed within the GH-J clan. Moreover, besides the effect of substrate entrance on its own, we strongly suggest that a highly conserved arginine residue (in the RDP motif) rather than the previously proposed Tyr motif (not conserved) provides the proton to increase the pKa of the acid-base catalyst.


[Prostatic carcinoma in a young adult: a case report].
We report a case of carcinoma of the prostate in a 30-year-old man. Serum acid phosphatase was normal. A transrectal biopsy of the prostate demonstrated an undifferentiated carcinoma. Total prostatocystectomy was performed and subsequent pathologic report stated that the mass was an undifferentiated carcinoma of the prostate gland. Metastases to the intrapelvic lymph node were present. Although immunohistochemical prostatic acid phosphatase (PAP) activity was not demonstrated, prostatic specific antigen (PSA) staining revealed a positive reaction within the tumor cells, confirming prostatic carcinoma. The patient's course has been uneventful without any recurrence by the intermittent adjuvant chemotherapy 8 months postoperatively. Review of the literature in Japan disclosed 16 cases (including our case) of carcinoma of the prostate in patients under 40 years of age.


Ritonavir and ixazomib kill bladder cancer cells by causing ubiquitinated protein accumulation.
There is no curative treatment for advanced bladder cancer. Causing ubiquitinated protein accumulation and endoplasmic reticulum stress is a novel approach to cancer treatment. The HIV protease inhibitor ritonavir has been reported to suppress heat shock protein 90 and increase the amount of unfolded proteins in the cell. If the proteasome functions normally, however, they are rapidly degraded. We postulated that the novel proteasome inhibitor ixazomib combined with ritonavir would kill bladder cancer cells effectively by inhibiting degradation of these unfolded proteins and thereby causing ubiquitinated proteins to accumulate. The combination of ritonavir and ixazomib induced drastic apoptosis and inhibited the growth of bladder cancer cells synergistically. The combination decreased the expression of cyclin D1 and cyclin-dependent kinase 4, and increased the sub-G1 fraction significantly. Mechanistically, the combination caused ubiquitinated protein accumulation and endoplasmic reticulum stress. The combination-induced apoptosis was markedly attenuated by the protein synthesis inhibitor cycloheximide, suggesting that the accumulation of ubiquitinated proteins played an important role in the combination's antineoplastic activity. Furthermore, the combination induced histone acetylation cooperatively and the decreased expression of histone deacetylases was thought to be one mechanism of this histone acetylation. The present study provides a theoretical basis for future development of novel ubiquitinated-protein-accumulation-based therapies effective against bladder cancer.


Accurate pseudoanalytical solution for steady-state biofilms.
An extremely accurate pseudoanalytical solution for the flux of substrate into a steady-state biofilm is developed. The standard deviations between the substrate fluxes computed from the pseudoanalytical solution and the numerical solution were less than 2.6%. Additional advantages of the pseudoanalytical solution are that it has no inaccuracies around S(min) (*) = 1 and it is composed of single continuous functions applicable to the whole S(min) (*) region.