Effects of androstenedione administration on epitestosterone metabolism in men.
Androstenedione is a steroid hormone sold over-the-counter to individuals who expect that it will enhance strength and athletic performance. Endogenous androstenedione is the immediate precursor of testosterone. To evaluate the metabolism of oral androstenedione, we randomly assigned 37 healthy men to receive 0 (group 1), 100 mg (group 2), or 300 mg (group 3) of androstenedione in a single daily dose for 7 days. Eight-hour urines were collected 1 day before the start of androstenedione, and on days 1 and 7. Using gas chromatography-mass spectrometry, we measured excretion rates of glucuronide-conjugated epitestosterone, its putative precursor (E-precursor), and metabolites (EM-1 and EM-2), and we evaluated possible markers of androstenedione administration. Day 1 and 7 rates were not different: the means were averaged. The means (microg/h) for groups 1, 2, and 3, respectively were, for epitestosterone 2.27, 7.74, and 18.0; for E-precursor, 2.9, 2.0, and 1.5; for EM-1/E-precursor 0.31, 1.25, and 2.88; for EM-2/E-precursor 0.14, 0.15, and 1.15; for testosterone/epitestosterone (T/E) 1.1, 3.5, and 3.2. Epitestosterone, EM-1, and EM-2 excretion was greater in groups 2 and 3 versus group 1 (0.0001 < P < 0.03), as were EM-1/E-precursor, EM-2/E-precursor, and T/E. E-precursor excretion was lower in groups 2 (P = 0.08) and 3 (P = 0.047) versus group 1. Androstenedione increases excretion of epitestosterone and its two metabolites, while decreasing that of its precursor. Elevated ratios of EM-1- and EM-2/E-precursor, and the presence of 6alpha-hydroxyandrostenedione are androstenedione administration markers.


Ankylosis of maxillary permanent first molar.
A case of ankylosis of the maxillary right first permanent molar has been described. For 19 months, no change had been observed clinically or radiographically in the development of the tooth. A diagnosis of ankylosis of the unerupted tooth was made. Instead of extracting the tooth, the ankylosed tooth was rocked in the bone to break the point of ankylosis. The tooth subsequently erupted and satisfactory functional results were obtained. It is suggested that this procedure should be attempted in other cases of ankylosis of permanent teeth.


Fetal viral myocarditis and congenital complete heart block in a pregnancy complicated by systemic lupus erythematosus. A case report.
Congenital complete heart block has been associated with collagen vascular disorders. Many etiologies have been proposed for the myocarditis that develops and results in the complete heart block. Transplacental antibody transfer--specifically, antibodies to tissue-soluble ribonuclear protein antigen-A--has been implicated as the etiology of the myocarditis. This case supports the association of this antibody while suggesting that viral infection may serve as an initiator of the autoimmune destruction. Also, fetal viral infection alone must be considered a possible cause of congenital complete heart block.


The Serial Duplex Index Improves Differential Diagnosis of Acute Renal Transplant Dysfunction.
Renal duplex sonography represents a standard noninvasive diagnostic procedure to demonstrate morphologic changes in acute kidney transplant dysfunction. We investigated whether a newly developed serial duplex index (SDI) can differentiate between acute cellular rejection and acute vascular rejection more effectively than the established Doppler parameters of the resistive index (RI) and pulsatility index (PI) in recently transplanted patients. Serial duplex scans of patients with histologically proven acute tubular necrosis (n = 25), acute cellular rejection (n = 28), acute vascular rejection (n = 18), and normal graft function (n = 50, partially protocol biopsied) were retrospectively analyzed. For each patient, the RI, PI, and cortex-pelvis proportion (CPP) were included from the day of biopsy (t0) and 3 to 7 days before biopsy (t-1). The sequential CPP ratio (CPPt0 /CPPt-1 ), RI ratio (RIt0 /RIt-1 ), and PI ratio (PIt0 /Pit-1 ) were determined. The SDI was calculated as: RI ratio × PI ratio/CPP ratio. The diagnostic accuracy of the SDI was compared with that of the RI and PI ratios. Selected groups were statistically comparable in all routinely determined transplant parameters. The SDI was significantly different between patients with normal graft function, acute cellular rejection, and acute vascular rejection (P < .01, analysis of variance on ranks), whereas the RI and PI ratios were only significantly different between patients with normal graft function and acute vascular rejection (P < .05, analysis of variance on ranks). The indices' ranges were defined by the 95% confidence intervals between the allograft functions. The developed SDI was able to detect acute renal transplant rejection with greater sensitivity and specificity than the RI and PI ratios. Since the SDI distinguishes between acute tubular necrosis, acute cellular rejection, and acute vascular rejection, it might be a supportive tool to indicate renal biopsy.


[Primary multiple tumors of the larynx].
A rare combination of two histogenetically different tumours of the larynx in a male patient of 43, namely chondrosarcoma and squamous cell carcinoma is described. These tumours developed simultaneously, therefore the carcinoma could not result from the radiotherapy. No similar observation has been published in the literature.


Integrating the humanities in the education of health professionals: implications for search and retrieval of information.
This article examines the increasing use of the humanities in the education of health professionals and posits that the approach may be of use in teaching health professionals information search and retrieval skills. However little evidence exists to support the educational effectiveness of using the humanities. This lack of evidence raises concerns about the costs of financing this approach to learning. These costs include the issue of copyright which cannot be ignored. While the humanities might provide a more attractive approach to teaching information search and retrieval skills, further research is needed to justify the costs of this approach to learning in more general terms and urgent attention to.


[Carcinosarcoma and spindle cell carcinoma of the bladder. A comparison of 2 cases with an immunohistochemical study].
These two cases of bladder tumours with an unusual histological appearance were observed at Hôpital Saint-Louis in 1988. They contained two cellular components: the usual epithelial type and a spindle cell type. Immunohistochemistry performed in order to identify the various cell contingents established the diagnoses of carcinosarcoma and spindle cell carcinoma and emphasised the differences and similarities between these two entities, which we believe can be differentiated.


Positron emission tomography quantification of serotonin transporter binding in medication-free bipolar disorder.
Bipolar disorder (BD) is associated with abnormalities in the serotonin transporter (5-HTT), but specific in vivo findings have been discrepant. Using positron emission tomography (PET) and [(11)C]DASB, we compared 5-HTT binding between unmedicated depressed BD subjects and healthy volunteers (HVs). 5-HTT binding in six brain regions was compared between 17 depressed, unmedicated BD subjects and 31 HVs, using the outcome measure of VT/fP (proportional to the total number of available transporters). Alternative outcome measures were examined as well. 47% of BD were BP I; and 65% reported a prior suicide attempt. 5-HTT binding (VT/fP ) did not differ between BD and HV groups considering six brain regions of interest simultaneously (P = 0.24). In contrast, alternative outcome measures (BPF*, BPP*, and BPND*) indicated lower binding in BD compared with HV across these six regions of interest (BPF*: P = 0.047; BPP*: P = 0.032; BPND*: P = 0.031). 5-HTT binding was unrelated to suicide attempt history, depression severity, bipolar subtype, or history of past substance use disorder. Choice of outcome measure strongly affects comparisons of serotonin transporter binding using PET with [(11)C]DASB. We do not find evidence of abnormal 5-HTT binding in bipolar depression using our primary outcome measure, VT /fP . However, we did observe lower 5-HTT binding in BD with alternative outcome measures that are frequently used with [(11)C]DASB. Relative merits and assumptions of different outcome measures are discussed. Evaluation in larger samples and during different mood states, including remission, is warranted.


Chemical communication in proteobacteria: biochemical and structural studies of signal synthases and receptors required for intercellular signalling.
Cell-cell communication via the production and detection of chemical signal molecules has been the focus of a great deal of research over the past decade. One class of chemical signals widely used by proteobacteria consists of N-acyl-homoserine lactones, which are synthesized by proteins related to LuxI of Vibrio fischeri and are detected by proteins related to the V. fischeri LuxR protein. A related marine bacterium, Vibrio harveyi, communicates using two chemical signals, one of which, autoinducer-2 (AI-2), is a furanone borate diester that is synthesized by the LuxS protein and detected by a periplasmic protein called LuxP. Evidence from a number of laboratories suggests that AI-2 may be used as a signal by diverse groups of bacteria, and might permit intergeneric signalling. These two families of signalling systems have been studied from the perspectives of physiology, ecology, biochemistry, and more recently, structural biology. Here, we review the biochemistry and structural biology of both acyl-homoserine-lactone-dependent and AI-2-dependent signalling systems.


Combined endoscopic and transcutaneous approach for immobile parotid stones.
Salivary stones in the parotid gland that are not amenable to endoscopic extraction can safely be removed by a combined, endoscopic-external approach. Limitations for endoscopic removability of salivary stones from the parotid gland are immobile stones in the deeper salivary duct system. A combined endoscopic and transcutaneous approach gives access to these stones with possible preservation of the parotid gland. A total of 10 consecutive patients were treated using the combined approach for the management of sialolithiasis of the parotid gland. The patients had previously undergone a sialendoscopy under local anesthesia and were proven to have large, immobile salivary stones in the salivary duct. In all patients, the procedure was successfully conducted with extraction of impacted stones. In 60% of the patients, a temporary stenting of the salivary duct was performed. All patients showed good clinical results with restored drainage of the salivary duct system and preserved gland function.


Synthesis of lipidated green fluorescent protein and its incorporation in supported lipid bilayers.
Herein we report a semisynthetic method of producing membrane-anchored proteins. Ligation of synthetic lipids with designed anchor structures to proteins was performed using native chemical ligation (NCL) of a C-terminal peptide thioester and an N-terminal cysteine lipid. This strategy mimics the natural glycosylphosphatidylinositol (GPI) linkage found in many natural membrane-associated proteins; however, the synthetic method utilizes simple lipid anchors without glycans. Synthetically lipidated recombinant green fluorescent protein (GFP) was shown to be stably anchored to the membrane, and its lateral fluidity was quantitatively characterized by direct fluorescence imaging in supported membranes. Circumventing the steps of purification from native cell membranes, this methodology facilitates the reconstitution of membrane-associated proteins.


Listening to bluetooth beacons for epidemic risk mitigation.
The ongoing COVID-19 pandemic let to efforts to develop and deploy digital contact tracing systems to expedite contact tracing and risk notification. Unfortunately, the success of these systems has been limited, partly owing to poor interoperability with manual contact tracing, low adoption rates, and a societally sensitive trade-off between utility and privacy. In this work, we introduce a new privacy-preserving and inclusive system for epidemic risk assessment and notification that aims to address these limitations. Rather than capturing pairwise encounters between user devices as done by existing systems, our system captures encounters between user devices and beacons placed in strategic locations where infection clusters may originate. Epidemiological simulations using an agent-based model demonstrate that, by utilizing location and environmental information and interoperating with manual contact tracing, our system can increase the accuracy of contact tracing actions and may help reduce epidemic spread already at low adoption.


Structural Insights into the TLA-3 Extended-Spectrum β-Lactamase and Its Inhibition by Avibactam and OP0595.
The development of effective inhibitors that block extended-spectrum β-lactamases (ESBLs) and restore the action of β-lactams represents an effective strategy against ESBL-producing Enterobacteriaceae We evaluated the inhibitory effects of the diazabicyclooctanes avibactam and OP0595 against TLA-3, an ESBL that we identified previously. Avibactam and OP0595 inhibited TLA-3 with apparent inhibitor constants (Kiapp) of 1.71 ± 0.10 and 1.49 ± 0.05 μM, respectively, and could restore susceptibility to cephalosporins in the TLA-3-producing Escherichia coli strain. The value of the second-order acylation rate constant (k2/K, where k2 is the acylation rate constant and K is the equilibrium constant) of avibactam [(3.25 ± 0.03) × 103 M-1 · s-1] was closer to that of class C and D β-lactamases (k2/K, <104 M-1 · s-1) than that of class A β-lactamases (k2/K, >104 M-1 · s-1). In addition, we determined the structure of TLA-3 and that of TLA-3 complexed with avibactam or OP0595 at resolutions of 1.6, 1.6, and 2.0 Å, respectively. TLA-3 contains an inverted Ω loop and an extended loop between the β5 and β6 strands (insertion after Ser237), which appear only in PER-type class A β-lactamases. These structures might favor the accommodation of cephalosporins harboring bulky R1 side chains. TLA-3 presented a high catalytic efficiency (kcat/Km ) against cephalosporins, including cephalothin, cefuroxime, and cefotaxime. Avibactam and OP0595 bound covalently to TLA-3 via the Ser70 residue and made contacts with residues Ser130, Thr235, and Ser237, which are conserved in ESBLs. Additionally, the sulfate group of the inhibitors formed polar contacts with amino acid residues in a positively charged pocket of TLA-3. Our findings provide a structural template for designing improved diazabicyclooctane-based inhibitors that are effective against ESBL-producing Enterobacteriaceae.


Caloric restriction and chronic inflammatory diseases.
A reduction in calorie intake [caloric restriction (CR)] appears to consistently decrease the biological rate of aging in a variety of organisms as well as protect against age-associated diseases including chronic inflammatory disorders such as cardiovascular disease and diabetes. Although the mechanisms behind this observation are not fully understood, identification of the main metabolic pathways affected by CR has generated interest in finding molecular targets that could be modulated by CR mimetics. This review describes the general concepts of CR and CR mimetics as well as discusses evidence related to their effects on inflammation and chronic inflammatory disorders. Additionally, emerging evidence related to the effects of CR on periodontal disease in non-human primates is presented. While the implementation of this type of dietary intervention appears to be challenging in our modern society where obesity is a major public health problem, CR mimetics could offer a promising alternative to control and perhaps prevent several chronic inflammatory disorders including periodontal disease.


Complementary therapy use among older rural adults.
Explore use, cost, and satisfaction with the quality and effectiveness of complementary therapy among older rural adults. Descriptive survey. A random sample of 325 older adults from rural communities throughout Montana and North Dakota. Participants were interviewed by telephone. Only 57 participants (17.5%) had used complementary providers and most sought this care for chronic problems, heard about providers through word-of-mouth information, and were satisfied with the care. A total of 35.7% (116) used self-directed complementary practices and most used these practices for health promotion, heard about them through informal sources, and found them to be at least somewhat helpful. Of the 325 participants, 45.2% (147) used some form of complementary care, e.g., providers, self-directed practices, or both. Participants used as much complementary care as is found in national studies. Most spent relatively little out-of-pocket for complementary care. Understanding the health care choices that older rural residents make, including complementary health care, is paramount for a comprehensive approach to meeting their health care needs.


Structural and comparative analysis of the mouse gene for pigment epithelium-derived factor (PEDF).
Pigment epithelium-derived factor (PEDF), a neurotrophic protein, is a member of the serine protease inhibitor supergene family. PEDF promotes both neuronal phenotype in cultured Y79 cells and neuronal survival in cultured cerebellar granulocytes. The purpose of this study was to clone the mouse PEDF gene and to determine its structure and levels of expression in different tissues. The mouse PEDF cDNA was cloned from a mouse liver cDNA library using human PEDF cDNA as a probe. The mouse PEDF gene was cloned from a mouse ES genomic P1 library. DNA sequencing was performed using a PE-Applied Biosystems model 373 automated fluorescent sequencer. The mouse PEDF cDNA is 1461 bp in length and contains an open reading frame of 417 amino acids. The mouse PEDF gene spans approximately 13 kb and, like the human, it is fragmented into 8 exons. The splice sites follow the AG/GT consensus rule. Southern blot analysis indicates that the mouse genome contains only one gene for PEDF. Northern blot analysis shows the presence of the PEDF transcript in a broad range of adult mouse tissues with liver showing the highest level of expression. The mouse and human PEDF promoters share overall 27% similarity but are nearly identical between mouse +86 to -166 and human +102 to -96. The present study will allow us to move from in vitro experiments to in vivo studies through the development of a "knock-out" mouse model.


[Lymphocytic interstitial pneumonia associated with Sjögren's syndrome and mediastinal lymphadenopathy: diagnosis by open-lung biopsy].
A 56-year-old woman was admitted to the hospital because of dry coughing and shortness of breath on exertion. In addition, dry eyes and cornea guttata suggested Sjögren's syndrome. Chest radiography revealed linear, reticular shadows throughout the lung fields, and enlargement of hilar and mediastinal lymph nodes. A specimen was obtained by transbronchial lung biopsy but the findings were not condusive; open-lung biopsy was done. The histopathological findings suggested lymphocytic meterstitial pneumonia. Results of genetic analysis and of immuno-histochemical examination conformed that the proliferating lymphocytes were polyclonal. Corticosteroids and immunosuppressive drugs have been used to treat lymphocytic interstitial pneumonia, and they were effective in this case.


Soil biochemical properties and stabilisation of soil organic matter in relation to deadwood of different species.
Despite the increasing number of studies on deadwood, we still have limited knowledge of its dynamics. The aim of this study was to examine the effect of deadwood on the biochemical properties of soil and stabilisation of soil organic matter (SOM). The investigation was carried out in the Czarna Rózga Reserve in Central Poland. The logs of four tree species at different stages of decomposition (III, IV and V) were selected for the analysis. Three replicate logs were sampled for each combination of decay classes, and the soil samples were collected from directly under the logs and from 1 m away from the logs. In this way, changes to the chemical and biochemical properties of the wood were determined. The SOM was physically fractioned. As the rate of deadwood decomposition increases, its biochemical activity increases and its chemical properties change. The biochemical activity, especially the soil's enzyme activity, was stimulated under highly decayed deadwood. The effects of deadwood are visible in SOM fractions, particularly in the content of the light fraction of SOM.


Smoking cessation and the effect of nicotine dependence on relapse rate in Izmir, Turkey.
Smoking is the leading preventable cause of death in the world. There is growing evidence of the need for community- based programs on smoking cessation. The main purpose of this study is to establish the rate of smoking cessation and restarting in 1 year at the Balçova Smoking Cessation Center. This is a prospective study with a study group of 359 individuals who quit smoking at the Balçova Smoking Cessation Center for at least 4 weeks between October 2009 and April 2010. The outcomes of the study were 1-year cessation rate and relapse rate. Individuals who reported restarting and/or had CO measurements above 6 ppm were accepted as quitters who had relapsed. The 1-year rate of smoking cessation was 30.1% for the study group. Of the subjects who quit smoking, 50.1% started smoking again during the 1-year follow-up. Relapse rate was also higher in nicotine addicts. Pharmacological treatment was associated with increased success rates in smoking cessation. Nicotine dependency was shown to be associated with lower rates of smoking cessation and higher rates of relapse. Therefore, it is important to begin smoking cessation attempts before individuals become serious addicts.


Beneficial Impact of Semicarbazide-Sensitive Amine Oxidase Inhibition on the Potential Cytotoxicity of Creatine Supplementation in Type 2 Diabetes Mellitus.
Creatine supplementation of the population with type 2 diabetes mellitus (T2DM) combined with an exercise program is known to be a possible therapy adjuvant with hypoglycemic effects. However, excessive administration of creatine leads to the production of methylamine which is deaminated by the enzyme semicarbazide-sensitive amine oxidase (SSAO) and as a result, cytotoxic compounds are produced. SSAO activity and reaction products are increased in the serum of T2DM patients. Creatine supplementation by diabetics will further augment the activity of SSAO. The current review aims to find a feasible way to ameliorate T2DM for patients who exercise and desire to consume creatine. Several natural agents present in food which are involved in the regulation of SSAO activity directly or indirectly are reviewed. Particularly, zinc-α2-glycoprotein (ZAG), zinc (Zn), copper (Cu), histamine/histidine, caffeine, iron (Fe), and vitamin D are discussed. Inhibiting SSAO activity by natural agents might reduce the potential adverse effects of creatine metabolism in population of T2DM.


Vibrational stark effect probes for nucleic acids.
The vibrational Stark effect (VSE) has proven to be an effective method for the study of electric fields in proteins via the use of infrared probes. To explore the use of VSE in nucleic acids, we investigated the Stark spectroscopy of nine structurally diverse nucleosides. These nucleosides contained nitrile or azide probes in positions that correspond to both the major and minor grooves of DNA. The nitrile probes showed better characteristics and exhibited absorption frequencies over a broad range; that is, from 2253 cm-1 for 2'-O-cyanoethyl ribonucleosides 8 and 9 to 2102 cm(-1) for a 13C-labeled 5-thiocyanatomethyl-2'-deoxyuridine 3c. The largest Stark tuning rate observed was |Deltamu| = 1.1 cm(-1)/(MV/cm) for both 5-cyano-2'-deoxyuridine 1 and N2-nitrile-2'-deoxyguanosine 7. The latter is a particularly attractive probe because of its high extinction coefficient (epsilon = 412 M-1cm-1) and ease of incorporation into oligomers.


Biventricular Interaction During Acute Left Ventricular Ischemia in Mice: A Combined In-Vivo and In-Silico Approach.
Computational models provide an efficient paradigm for integrating and linking multiple spatial and temporal scales. However, these models are difficult to parameterize and match to experimental data. Recent advances in both data collection and model analyses have helped overcome this limitation. Here, we combine a multiscale, biventricular interaction model with mouse data before and after left ventricular (LV) ischemia. Sensitivity analyses are used to identify the most influential parameters on pressure and volume predictions. The subset of influential model parameters are calibrated to biventricular pressure-volume loop data (n = 3) at baseline. Each mouse underwent left anterior descending coronary artery ligation, during which changes in fractional shortening and RV pressure-volume dynamics were recorded. Using the calibrated model, we simulate acute LV ischemia and contrast outputs at baseline and in simulated ischemia. Our baseline simulations align with the LV and RV data, and our predictions during ischemia complement recorded RV data and prior studies on LV function during myocardial infarction. We show that a model with both biventricular mechanical interaction and systems-level cardiovascular dynamics can quantitatively reproduce in-vivo data and qualitatively match prior findings from animal studies on LV ischemia.


Results of laparoscopic appendectomies performed with the use of titanium clips for closure of the appendicular stump.
Nowadays laparoscopy is used frequently not only in elective surgery but also in abdominal emergencies, including acute appendicitis. There are several techniques used to close the appendicular stump during laparoscopic appendectomy. The aim of the study was to present the results of minimally invasive appendectomies performed with the use of titanium clips. Patients operated on laparoscopically for acute appendicitis with the application of titanium clips between October 2012 and December 2013 were included in the study. We reviewed retrospectively patients` data including: age, sex, duration of the surgical procedure and hospital stay, mortality, intraoperative and postoperative complication rates. There were 93 patients (mean age=33.8 years, SD=15.23) in the analyzed group, including 60 men (mean age=33.5 years, SD= 15.07) and 33 women (mean age=33.9 years SD=15.26). The average duration of the surgical procedure was 66 min (SD= 33.15). The average length of hospital stay was 3.38 days (SD=1.62). No intraoperative complications were observed in the analyzed group. Post-operative complication rate was low (6 cases, 6.5%). No mortality was observed. Laparoscopic appendectomy with the application of titanium clips for closure of the appendicular stump is safe, associated with low complication rates and should be considered as a routine technique in everyday surgical practice.


Spatially distributed morphogen production and morphogen gradient formation.
Partial diferential equations and auxiliary conditions governing the activities of the morphogen Dpp in Drosophila wing imaginal discs were formulated and analyzed as Systems B, R, and C in [7] [9] [10]. All had morphogens produced at the border of anterior and posterior chamber of the wing disc idealized as a point, line, or plane in a one-, two-, or three-dimensional model. In reality, morphogens are synthesized in a narrow region of finite width (possibly of only a few cells) between the two chambers in which diffusion and reversible binding with degradable receptors may also take place. The present investigation revisits the extracellular System R, now allowing for a finite production region of Dpp between the two chambers. It will be shown that this more refined model of the wing disc, designated as System F, leads to some qualitatively diferent morphogen gradient features. One significant diference between the two models is that System F impose no restriction on the morphogen production rate for the existence of a unique stable steady state concentration of the Dpp-receptor complexes. Analytical and numerical solutions will be obtained for special cases of System F. Some applications of the results for explaining available experimental data (to appear elsewhere) are briefly indicated. It will also be shown how the effects of the distributed source of System F may be aggregated to give an approximating point source model (designated as the aggregated source model or System A for short) that includes System R as a special case. System A will be analyzed in considerable detail in [6], and the limitation of System R as an approximation of System F will also be delineated there.


Fluorescence energy transfer studies on the active site of papain.
Measurements have been performed of the excited-state lifetimes and fluorescence yields of papain tryptophan units when acyl derivatives of Phe-glycinal are bound at the active site of the enzyme. The enhancement of tryptophan fluorescence in complexes of papain with the acetyl or benzyloxycarbonyl derivatives is not stereospecific with respect to the configuration of the phenylalanyl residue, and the L and D isomers are equally effective as active-site-directed inhibitors of papain action. Evidence is offered in favor of the conclusion that this enhancement is primarily a consequence of the interaction of the phenylalanyl side chain of the inhibitor with Trp-69 of the enzyme. This residue can exchange fluorescence energy with the other four tryptophans of papain (Trp-7, Trp-26, Trp-177, Trp-181) upon excitation near their absorption maxima, but such "homotransfer" is absent if they are excited at the long-wave edge of their absorption spectra. Crystallographic data indicate that Trp-26 is most favorably positioned for efficient energy exchange with Trp-69, and the fluorescence data have been used to calculate a distance of 11 A between the two residues; this value is in satisfactory agreement with that found by crystallography. When derivatives of Phe-glycinal bearing an amino-terminal mansyl [6-(N-methylanilino)-2-naphthalene sulfonyl] group are bound at the active site of papain, the tryptophan fluorescence is quenched, as compared with that of the complex of papain with acetyl-Phe-glycinal, indicating energy transfer from papain tryptophan (most probably via Trp-26) to the fluorescent probe group. Although the L and D isomers of mansyl-Phe-glycinal are equally effective as inhibitors of papain action, the fluorescence quenching by the two isomers is different.


High risk of developing subsequent epilepsy in patients with sleep-disordered breathing.
Sleep-disordered breathing (SDB) is often associated with other medical disorders. Whether SDB interacts with other factors for developing subsequent epilepsy remains unclear. This population-based cohort study was conducted using the National Health Insurance Research Database of Taiwan. Patients aged >20 years and diagnosed with SDB between 2000 and 2010 comprised the SDB cohort (n = 138,507), and their data were compared with those of the comparison cohort (n = 138,507). The adjusted hazard ratio (aHR) for epilepsy was calculated using a multivariate Cox proportional hazards model. The SDB cohort had an increased risk of epilepsy (aHR = 1.50, 95% confidence interval [CI] = 1.36-1.66). The sex-stratified analysis revealed a significant adjusted hazard ratio (aHR) for epilepsy with a 1.51-fold higher risk for female patients, and also a significantly 1.49-fold higher risk for male patients in the SDB cohort. Although epilepsy incidence increased with age in both cohorts, different age groups in the SDB cohort all had a significantly higher risk of developing epilepsy than comparison cohort. This population-based cohort study indicates that patients with SDB are at a high risk of developing subsequent epilepsy, in both sexes and all age groups.


Super-resolution ultrasound localization microscopy based on a high frame-rate clinical ultrasound scanner: an in-human feasibility study.
Non-invasive detection of microvascular alterations in deep tissuesin vivoprovides critical information for clinical diagnosis and evaluation of a broad-spectrum of pathologies. Recently, the emergence of super-resolution ultrasound localization microscopy (ULM) offers new possibilities for clinical imaging of microvasculature at capillary level. Currently, the clinical utility of ULM on clinical ultrasound scanners is hindered by the technical limitations, such as long data acquisition time, high microbubble (MB) concentration, and compromised tracking performance associated with low imaging frame-rate. Here we present a robust in-human ULM on a high frame-rate (HFR) clinical ultrasound scanner to achieve super-resolution microvessel imaging using a short acquisition time (<10 s). Ultrasound MB data were acquired from different human tissues, including a healthy liver and a diseased liver with acute-on-chronic liver failure, a kidney, a pancreatic tumor, and a breast mass using an HFR clinical scanner. By leveraging the HFR and advanced processing techniques including sub-pixel motion registration, MB signal separation, and Kalman filter-based tracking, MBs can be robustly localized and tracked for ULM under the circumstances of relatively high MB concentration associated with standard clinical MB administration and limited data acquisition time in humans. Subtle morphological and hemodynamic information in microvasculature were shown based on data acquired with single breath-hold and free-hand scanning. Compared with contrast-enhanced power Doppler generated based on the same MB dataset, ULM showed a 5.7-fold resolution improvement in a vessel based on a linear transducer, and provided a wide-range blood flow speed measurement that is Doppler angle-independent. Microvasculatures with complex hemodynamics can be well-differentiated at super-resolution in both normal and pathological tissues. This preliminary study implemented the ultrafast in-human ULM in various human tissues based on a clinical scanner that supports HFR imaging, indicating the potentials of the technique for various clinical applications. However, rigorous validation of the technique in imaging human microvasculature (especially for those tiny vessel structure), preferably with a gold standard, is still required.


Effects of garlic oil (allium sativum) on acetic acid-induced colitis in rats: Garlic oil and experimental colitis.
Inflammatory bowel disease (IBD) is an important health problem. The most important hypotheses for the pathogenesis of this disease are the deterioration of immune responses and loss of tolerance against bacteria in the enteric flora. Although IBD has been widely investigated, its treatment remains difficult. This study aims to investigate the effects of garlic oil (GO) on an experimental colitis model. Twenty-eight rats were randomly divided into four equal groups as follows: group 1 (sham), group 2 (control), group 3 (topical treatment) and group 4 (topical and systemic treatment). An acetic acid-induced colitis model was produced in groups 2, 3 and 4 and was administered normal saline, topical GO and topical and systemic GO, respectively. Hydroxyproline levels were lower in the treatment groups than in the control group. TNF-α levels were significantly lower in group 3 than in group 2. Macroscopic scores were significantly lower in group 4 than in group 2. Significant differences were observed between the treatment and control groups according to their epithelial loss. GO can reduce colonic damage and inflammation in the acetic acid-induced colitis model, with effects on both local and systemic treatments, but with a more pronounced effect in local treatment.


Phase transitions of the Ashkin-Teller model including antiferromagnetic interactions on a type of diamond hierarchical lattice.
Using the real-space renormalization-group transformation, we study the phase transitions of the Ashkin-Teller model including the antiferromagnetic interactions on a type of diamond hierarchical lattices, of which the number of bonds per branch of the generator is odd. The isotropic Ashkin-Teller model and the anisotropic one are, respectively, investigated. We find that the phase diagram, for the isotropic Ashkin-Teller model, consists of five phases, two of which are associated with the partially antiferromagnetic ordering of the system, while the phase diagram, for the anisotropic Ashkin-Teller model, contains 11 phases, six of which are related to the partially antiferromagnetic ordering of the system. The correlation length critical exponents and the crossover exponents are also calculated.


Outcomes associated with a transcarotid artery revascularization-centered protocol in high-risk carotid revascularizations using the ENROUTE neuroprotection system.
This investigation describes the perioperative and early follow-up results associated with transcarotid artery revascularization (TCAR) in patients not participating in the Safety and Efficacy Study for Reverse Flow Used During Carotid Artery Stenting Procedure II (ROADSTER-2) registry using the ENROUTE neuroprotection system (ENPS; Silk Road Medical, Sunnyvale, Calif). A retrospective review was performed capturing all TCAR/ENPS procedures in patients deemed to be at high risk for complications after traditional carotid endarterectomy. All patients enrolled in the ROADSTER-2 registry were excluded, leaving only those treated outside trial regulations for analysis. Preoperative demographics, intraoperative variables, and perioperative and follow-up outcomes were abstracted and reported herein. From December 2015 to January 2018, there were 75 carotid arteries treated at our institution. All interventions were performed on carotid arteries that were symptomatic with ≥50% stenosis (46.7%) or asymptomatic with ≥80% stenosis (53.3%) by duplex ultrasound and computed tomography angiography. Technical success in our series was 97.3% (73/75), with treatment failures attributed to one case of common carotid artery dissection and another secondary to stent maldeployment in the external carotid artery. Perioperative (30-day) ipsilateral stroke rate was 2.7% (n = 2), myocardial infarction incidence was 0%, and mortality rate was 2.7% (n = 2). We did not observe any cranial nerve injuries. After a mean follow-up of 8.0 ± 6.7 months, no carotid stents required reintervention. However, we noted one instance of minor (<50%) in-stent stenosis and one asymptomatic stent thrombosis. One additional ipsilateral stroke was observed on follow-up, probably from a cardiac source. We report that dynamic reverse-flow TCAR using the ENPS continues to be safe, feasible, and efficacious with minimal risks of postoperative stroke, myocardial infarction, and mortality outside of ROADSTER-2 regulations.


Pulp Inflammation Diagnosis from Clinical to Inflammatory Mediators: A Systematic Review.
Similar to other tissues, the dental pulp mounts an inflammatory reaction as a way to eliminate pathogens and stimulate repair. Pulp inflammation is prerequisite for dentin pulp complex repair and regeneration; otherwise, chronic disease or pulp necrosis occurs. Evaluation of pulp inflammation severity is necessary to predict the clinical success of maintaining pulp vitality. Clinical limitations to evaluating in situ inflammatory status are well-described. A molecular approach that aids clinical distinction between reversible and irreversible pulpitis could improve the success rate of vital pulp therapy. The aim of this article is to review inflammatory mediator expression in the context of clinical diagnosis. We searched PubMed and Cochrane databases for articles published between 1970 and December 2016. Only published studies of inflammatory mediator expression related to clinical diagnosis were eligible for inclusion and analysis. Thirty-two articles were analyzed. Two molecular approaches were described by study methods, protein expression analysis and gene expression analysis. Our review indicates that interleukin-8, matrix metalloproteinase 9, tumor necrosis factor-α, and receptor for advanced glycation end products expression increase at both the gene and protein levels during inflammation. Clinical irreversible pulpitis is related to specific levels of inflammatory mediator expression. The difference in expression between reversible and irreversible disease is both quantitative and qualitative. On the basis of our analysis, in situ quantification of inflammatory mediators may aid in the clinical distinction between reversible and irreversible pulpitis.


Effect of training status on beta-range corticomuscular coherence in agonist vs. antagonist muscles during isometric knee contractions.
Antagonist muscle co-activation is thought to be partially regulated by cortical influences, but direct motor cortex involvement is not fully understood. Corticomuscular coherence (CMC) measures direct functional coupling of the motor cortex and muscles. As antagonist co-activation differs according to training status, comparison of CMC in agonist and antagonist muscles and in strength-trained and endurance-trained individuals may provide in-depth knowledge of cortical implication in antagonist muscle co-activation. Electroencephalographic and electromyographic signals were recorded, while 10 strength-trained and 11 endurance-trained participants performed isometric knee contractions in flexion and extension at various torque levels. CMC magnitude in 13-21 and 21-31 Hz frequency bands was quantified by CMC analysis between Cz electroencephalographic electrode activity and all electromyographic signals. CMC was significant in both 13-21 and 21-31 Hz frequency bands in flexor and extensor muscles regardless of participant group, torque level, and direction of contraction. CMC magnitude decreased more in antagonist than in agonist muscles as torque level increased. Finally, CMC magnitude was higher in strength-trained than in endurance-trained participants. These findings provide experimental evidence that the motor cortex directly regulates both agonist and antagonist muscles. Nevertheless, the mechanisms underlying muscle activation may be specific to their function. Between-group modulation of corticomuscular coherence may result from training-induced adaptation, re-emphasizing that corticomuscular coherence analysis may be efficient in characterizing corticospinal adaptations after long-term muscle specialization.


The occupational roles of women with anorexia nervosa.
This study's objective was to understand how occupational roles of individuals with anorexia nervosa are configured. The sample was composed of a control group and 11 adult women with anorexia nervosa being cared for by the Eating Disorders Care Group in a hospital in Ribeirão Preto, SP, Brazil. Socio-demographic and anthropometric data were collected and the Role Checklist was applied. The results revealed a significant loss of roles for women with anorexia nervosa in relation to the performance of the roles worker, friend, and amateur/hobbyist, supporting the idea that psychosocial harm may arise from this eating disorder. The evaluation of occupational roles in the treatment of eating disorders is an important strategy for planning Occupational Therapy activities and supporting the creation of healthier spaces to enable individuals to resume occupational roles, and acquire independence and autonomy.


[Heart rate recovery after exercise in subjects undergoing cardiologic rehabilitation].
Research indicates that slow heart rate recovery 1-2 minutes after exercise is a predictor of cardiovascular mortality, sudden mortality as well. It is hardly related to myocardial ischemia; most of all, it is related to impaired activities of the parasympathetic system. The purpose of this study was to assess HRR in subjects after a surgical treatment (percutaneous coronary intervention - PCI) of acute coronary syndromes (ACS), undergoing cardiologic rehabilitation. The study comprised 58 subjects, 42 men and 16 women aged 49-68 (56.8 +/- 7.6) after ACS treated with PCI undergoing cardiologic rehabilitation (stage 2)--group I. The comparative group comprised 34 subjects, 25 men and 9 women aged 46-61 (55.5 +/- 8.9 lat) who were clinically healthy and who underwent a single sub-maximal exercise test--group II. In subjects undergoing rehabilitation, an exercise test was performed twice --before and after stage 2 of rehabilitation. HRR was defined as a difference between the peak heart rate and those after the 1st (HRR1) and 2nd (HRR) minute of recovery. After stationary rehabilitation as a part of stage 2 of cardiologic rehabilitation, HRR2 was 26.3 +/- 10.6/min and was 40.8/min +/- 13.8/min; both these values were not significantly different from values observed in healthy subjects (p > 0.05). Significant increase in HRR1 and HRR2 was observed after the completion of stage 2 of cardiologic rehabilitation in ill subjects (p < 0.05) as compared to initial values. In subjects after ACS treated with PCI, it is observed that HRR changes back to normal values observed in healthy subjects as a result of underwent cardiologic rehabilitation. HRR should become an important factor in assessing effectiveness of conducted cardiologic rehabilitation.


[A study of knowledge, attitude and health behavior toward tuberculosis among non-immigrant Korean people in Japan].
Knowledge, attitude and health behavior toward tuberculosis among Non-immigrant Korean people in Japan was researched by using questionnaire because of increasing the number of the tuberculosis patients among those group. The Korean member of protestant churches in Tokyo were subjects for the survey. Immigrant Korean people and their descendants were excluded. The questionnaire form was written in Korean language under the guidance of native Korean tuberculosis specialists. Proportion of Response was 53.1%, or 251 among 473 from 10th January to 30th June in 1992. The knowledge of tuberculosis among them was revealed to be higher than among ordinary native Korean people. It was different statistically by generation, namely, younger subjects aged less than 40 years old tended to answer that tuberculosis was a minor illness. The mass screening system in Japan was well known by the subjects, as shown by the fact that 72.4% of them answered that they knew about it. But only 56.6% of them replied that they actually took the mass screening. The source of its information was different statistically by sex, occupation, and generation. As for their health behavior, nearly two third (63.7%) of them visited the hospital or dispensary quickly when they fell sick. A small number of them answered that they could not visit a doctor because of their problems with the Japanese language. More than 80% of them possessed a National Health Insurance certificate. This proportion varied according to the period of stay in Japan. That is to say, The group which stayed in Japan less than one year was significantly the lowest because they were limited in their ability to enter National Health Insurance.


Facilitating consumer clinical information seeking by maintaining referential context: evaluation of a prototypic approach.
Millions of consumers seek health information on the Internet. Unfortunately, this searching often falls short because of design limitations of many consumer-oriented Web sites. In this paper, we describe an approach that addresses several known barriers to consumer health information seeking. This approach primarily involves maintaining the referential context throughout a consumer's search for information. To maintain referential context, this approach uses multiple levels of hierarchical constructs to organize complex information, and data elements are toggled to minimize the need for scrolling. An information resource based on this approach was implemented for information about smoking using standard Web technologies. The resource was evaluated by 31 diverse consumers through standardized usability instruments. Consumers found the resource to be easy to navigate and to use. We conclude that the approach described in this manuscript could be applied more broadly to facilitate the organization and presentation of consumer health information.


Genetic variation analysis and relationships among environmental strains of Scedosporium apiospermum sensu stricto in Bangkok, Thailand.
The Scedosporium apiospermum species complex is an emerging filamentous fungi that has been isolated from environment. It can cause a wide range of infections in both immunocompetent and immunocompromised individuals. We aimed to study the genetic variation and relationships between 48 strains of S. apiospermum sensu stricto isolated from soil in Bangkok, Thailand. For PCR, sequencing and phylogenetic analysis, we used the following genes: actin; calmodulin exons 3 and 4; the second largest subunit of the RNA polymerase II; ß-tubulin exon 2-4; manganese superoxide dismutase; internal transcribed spacer; transcription elongation factor 1α; and beta-tubulin exons 5 and 6. The present study is the first phylogenetic analysis of relationships among S. apiospermum sensu stricto in Thailand and South-east Asia. This result provides useful information for future epidemiological study and may be correlated to clinical manifestation.


The immunological personality of close relatives of SLE patients.
Immunological abnormalities seen in relatives of patients with autoimmune disorders can be useful in understanding the pathogenesis of the disease since, unlike in patients, they cannot result from the disease process or drug treatment. In this article we present a brief overview of our studies of the basic immunological status of close relatives of SLE patients. We looked at blood levels of IgG, IgM and antibodies to double-stranded DNA, as well as at NK cell numbers and cytotoxic activity and the levels of NKT, B and T cells. As many as 60% of relatives showed one or more abnormalities in these assays. Most notably there were increased levels of IgG in male and female relatives and a reduction of IgM in females. IgG correlated inversely with NKT cell numbers adding strength to the concept that the presence of IgG autoantibodies in patients is due to impaired regulation by NKT cells. IgM, on the other hand, correlated inversely with NK cells which may thus have a role in bringing about the reduced IgM seen in some patients. Immunological abnormalities were found to be more often associated with parents and offspring of patients than with their siblings, pointing to the involvement of environmental or epigenetic influences in lupus pathogenesis.


[Impaired cerebral autoregulation in asymptomatic patients with carotid artery stenosis: comparison of acetazolamide-SPECT and transcranial CO(2)-dopplersonography].
The indication for operation in patients with asymptomatic high-grade carotid artery stenosis is still under debate. Since impaired cerebrovascular autoregulation is associated with an increased risk for ischaemic events, assessment of cerebral vascular reactivity might be a valuable selection criterion for surgery. The aim of our study was therefore to evaluate the incidence of impaired autoregulation in asymptomatic patients with acetazolamide-single photon emission computed tomography (ACZ-SPECT) and transcranial CO (2)-dopplersonography (CO (2)-TCD). Furthermore, both methods were compared in regard to results and clinical practicability. In 42 patients with high-grade (> 70 %) asymptomatic carotid artery stenosis, cerebral perfusion and vascular reactivity were assessed with resting and ACZ-enhanced SPECT scans. In 31 of these patients the CO (2) reactivity of cerebral perfusion was determined by TCD and expressed as normalized autoregulation reserve (NAR). Cerebral perfusion was decreased in 14.3 %. In ACZ-SPECT 26 % and in CO (2)-TCD 28 % revealed an impaired vascular reactivity. Conformity of both methods was high (kappa = 0.93). TCD was superior in practicability, but only applicable in 81 % due to a missing temporal bone window for insonation. In accordance ACZ-SPECT and CO (2)-TCD could detect impaired vascular reactivity in a quarter of asymptomatic patients. Both TCD and SPECT could be of value for preoperative selection in this group of patients, whereby sonography is recommended for daily diagnostic work-up.


Team climate and attitudes toward information and communication technology among nurses on acute psychiatric wards.
Objectives: To describe the association of team climate with attitudes toward information and communication technology among nursing staff working on acute psychiatric wards. Background: Implementation of ICT applications in nursing practice brings new operating models to work environments, which may affect experienced team climate on hospital wards. Method: Descriptive survey was used as a study design. Team climate was measured by the Finnish modification of the Team Climate Inventory, and attitudes toward ICT by Burkes' questionnaire. The nursing staff (N = 181, n = 146) on nine acute psychiatric wards participated in the study. Results: It is not self-evident that experienced team climate associates with attitudes toward ICT, but there are some positive relationships between perceived team climate and ICT attitudes. The study showed that nurses' motivation to use ICT had statistically significant connections with experienced team climate, participative safety (p = 0.021), support for innovation (p = 0.042) and task orientation (p = 0.042). Conclusion: The results suggest that asserting team climate and supporting innovative operations may lead to more positive attitudes toward ICT. It is, in particular, possible to influence nurses' motivation to use ICT. More attention should be paid to psychosocial factors such as group education and co-operation at work when ICT applications are implemented in nursing.


Glutamic acid 274 is the nucleophile in the active site of a "retaining" exoglucanase from Cellulomonas fimi.
In addition to its known substrate activity with p-nitrophenyl beta-cellobioside, the exoglucanase from Cellulomonas fimi has been shown to utilize substituted phenyl beta-glucosides as substrates, of which the best is 2',4'-dinitrophenyl beta-D-glucopyranoside. The enzyme can be inactivated by treatment with 2',4'-dinitrophenyl 2-deoxy-2-fluoro-beta-D-glucopyranoside, by trapping of the covalent intermediate in catalysis, as has been shown for a beta-glucosidase (Withers, S.G., and Street, I.P. (1988) J. Am. Chem. Soc. 110, 8551-8553). The intermediate formed is stable but can undergo turnover in the presence of cellobiose, reactivating the enzyme by transglycosylation. Using a tritium-labeled inactivator it has been possible to isolate and sequence a radiolabeled peptide from this enzyme, and the active site nucleophile has been identified as glutamic acid residue 274. This glutamic acid residue and its sequentially proximal amino acids are absolutely conserved in the homologous family F of cellulases.


[Therapeutic effects of a new taurine derivative in chronic hepatic diseases].
Therapeutic properties of a novel taurin N-phenylakyl derivative (TAU-15) in a dose 25 mg/kg were studied in male rats on models of chronic toxic hepatitis and partial hepatectomy. A hepatoprotective action of TAU-15 was established which is due to antioxidative action of TAU-15 and its ability to normalize protein-synthetizing activity of hepatocytes.


Long-term results after repeated surgical removal of pulmonary metastases.
Although surgical resection is accepted widely as first-line therapy for pulmonary metastases, few data exist on the surgical treatment of recurrent pulmonary metastatic disease. In a retrospective study, we analyzed patients who were operated on repeatedly for recurrent metastatic disease of the lung with curative intent over a 20-year period. From 1973 to 1993, 396 metastasectomies were performed in 330 patients. The study population included patients with any histologic tumor type who had undergone at least two (range, 2 to 4) complete surgical procedures because of recurrent metastatic disease. Surgical and functional resectability of the recurrent lung metastases and control of the primary lesion served as objective criteria for reoperation. A subgroup of 35 patients that included patients with histologic findings such as epithelial cancer and osteosarcoma then was analyzed retrospectively to calculate prognosis and define selection criteria for repeated pulmonary metastasectomy. The 5- and 10-year survival rates after the first metastasectomy were 48% and 28%, respectively. The overall median survival was 60 months. A mean disease-free interval (calculated for all intervals, with a minimum of two) of greater than 1 year was significantly associated with a survival advantage beyond the last operation. Univariate analysis failed to show size, number, increase or decrease in number or size, or distribution of metastases as factors related significantly to survival. Although patients with different histologic tumor types were included, the study population appeared to be homogeneous in terms of survival benefit and prognostic factors, and it probably represented the selection of biologically favorable tumors in which histology, size, number, and laterality are of minor importance. We conclude that patients who are persistently free of disease at the primary location but who have recurrent, resectable metastatic disease of the lung are likely to benefit from operation a second, third, or even fourth time.


Shedding light on cashmere goat hair follicle biology: from morphology analyses to transcriptomic landascape.
Cashmere goat is known for its precious undercoat. Being photoperiod-dictated, cashmere growth has been studied focusing mainly on hair follicle cycle phases (anagen, catagen and telogen). An accurate molecular knowledge of the goat hair follicle cycle, disentangling gene expression changes during phases and recognizing timing boundaries, could be useful to improve cashmere goat management and ultimately cashmere production. To better describe goat's hair follicle transcriptome we applied RNA-sequencing to isolated hair follicles from five Italian cashmere goats, during the anagen and catagen phase, identifying total of 214 differentially expressed genes (DEGs): 97 were up-regulated while 117 were down-regulated in catagen with respect to anagen. Gene Ontology and pathway analysis were performed. We detected 144 significant pathways spanning from estrogen, pluripotency of stem cells, thermogenesis and fatty acid metabolism that were strongly expressed during the hair follicle phases analysed. Finally, we validated promising DEGs by RT-qPCR in the same set of samples as well as in hair follicles and entire skin biopsies of another cashmere goats cohort accounting for early anagen, anagen, early catagen, and catagen phases. As in the isolated hair follicles, some target genes were homogenously modulated during the four hair follicle phases. Ceruloplasmin (CP) and Keratin 4 (K4), confirmed their clear cut expression between growing and resting phase. In fact, K4 was almost absent in catagen phases while CP was barely expressed in anagen phases. In particular, the strong expression of K4 in early anagen makes it an eligible marker to track the beginning of a new hair cycle, and therefore defining the optimum time for cashmere harvesting.


Expression and prognostic significance of cyclin D3 in ovarian adenocarcinomas.
Abnormal expression of cell cycle regulators may contribute to malignant transformation. However, the clinical significance of the expression of cyclin D3 in ovarian cancer remains undefined. We therefore conducted a retrospective investigation to address the role of this cell-cycle protein in this tumor. In our study, paraffin-embedded tissue from 109 nonbenign epithelial ovarian tumors, including 17 tumors of low malignant potential and 92 primary adenocarcinomas, was stained immunohistochemically for cyclin D3. Most of the cases had previously been stained for pRb, p21Cip1, p27Kip1, p53, and Ki-67 antigen. Expression of cyclin D3 was correlated with clinicopathologic features, the expression of other cell cycle regulators, and postoperative survival of patients. Cyclin D3 levels were significantly higher in tumors of low malignant potential than in adenocarcinomas (P = 0.0002). In the latter group, cyclin D3 expression decreased with increasing grade (P = 0.0004) and advancing stage (P = 0.0315). Cyclin D3 expression positively correlated with pRb, p21Cip1, and p27Kip1 levels (P = 0.0021; P = 0.0036; P < 0.0001, respectively) and negatively with p53 and Ki-67 (P = 0.0003; P < 0.0001). Absent cyclin D3 expression was an important indicator of poor survival in univariate analysis in the entire cohort (P > 0.00010) and in the platinum-treated patients (P = 0.001) and in multivariate analysis (P = 0.044). Our results demonstrate that absent or decreased cyclin D3 expression is adversely related to several clinicopathologic indicators of aggressiveness in ovarian adenocarcinomas and is combined with a better prognosis, suggesting that cyclin D3 may have a biological role distinct from that of other G1 cyclins which is possibly regulated through interaction with other cell cycle genes.


Mapping of internal mammary vessels by multidetector computed tomography for parasternal transthoracic intervention guidance.
To measure the distances between the internal mammary arteries (IMAs) and the sternum for determining a route for parasternal transthoracic interventions. Coronal reconstructed multidetector computed tomography images of 202 patients were evaluated to measure the distances between the IMAs and sternum at 3 levels (manubrium, midsternal corpus, and distal sternal corpus). The arrangements (2 veins-single artery or single vein-single artery) were documented. The most frequently encountered arrangement (42.1%) was a single vein and single artery on both sides. Mean distances on the right and left sides were 10.2 and 10.1 cm at the manubrium, 14.5 and 14.3 cm at the midsternal corpus, and 17.9 and 17.5 cm at distal sternal corpus, respectively. To be aware of the courses of the IMAs at different sternal levels is important during attempts at transthoracic intervention via a parasternal approach.


A failure of mucocutaneous lymphangiogenesis may underlie the clinical features of lipoid proteinosis.
Lipoid proteinosis (LiP) (OMIM 247100) is a rare autosomal recessive disease caused by loss of function mutations in the extracellular matrix protein 1 gene, ECM1, on chromosome 1q21. LiP is characterized clinically by hoarseness in early infancy, followed by waxy papules and plaques on the face and body along with pox-like and acneiform scars. We studied a 20-year-old Japanese woman with LiP. She was born of consanguineous parents. Biopsy specimens obtained from a nodule on the elbow were used for histopathology, immunohistology and electron microscopy. Exons 6 and 7 of ECM1 were amplified by polymerase chain reaction (PCR) from genomic DNA from the proband, her parents, her brother and an unrelated person. PCR products were sequenced to detect the mutation. Histopathological examination revealed an irregular mass of calcium beneath deposits of a hyaline material in the dermis. Immunofluorescence double staining showed that the CD31-positive microvascular density was increased but that staining for the lymphatic-specific hyaluronan receptor LYVE-1 was drastically diminished in lesional compared with nonlesional skin of the patient and with normal skin. Electron microscopy revealed marked concentric reduplication of basal laminae not only around blood vessels but also around solitary dermal cells positive for Weibel-Palade bodies scattered in the hyaline material. Sequencing of the PCR products revealed a homozygous frameshift mutation, 507delT, in exon 6. This led to a premature stop codon 23 bp downstream. The results of immunopathological and ultrastructural characterization suggest that a failure of mucocutaneous lymphangiogenesis may underlie the clinical features of LiP. Identification of mutation 507delT in a Japanese patient with LiP further supports the thesis that this mutation represents a recurrent mutation in ECM1 in patients with LiP. To our knowledge, this case represents the first report of calcinosis cutis occurring in LiP.


Hydrodynamic and static performance evaluation of the moving-actuator type biventricular assist device, AnyHeart.
This study evaluated the hydrodynamic characteristics and efficiency of the moving-actuator type implantable biventricular assist device (BVAD), AnyHeart. A blood analog made of 52% glycerin and 48% water was used to simulate the density and viscosity of blood. The maximum pump flow was 9 L/min with 28.8 watts of power input, and the maximum electrical-to-hydraulic power conversion efficiency was approximately 11% at a pump flow of 3.5 L/min. The pump was able to generate 4 L/min output against 100 mm Hg afterload with less than 9 watts of power input. In addition to the overall system efficiency, the inner subpart power conversion efficiency was also evaluated. The system was subdivided according to system mechanism into three major parts: motor part, actuator part, and blood sac part. In normal working conditions (4 L/min, 100 mm Hg) with the AnyHeart, the total system efficiency was 8%, with subpart efficiencies of 50%, 85%, and 19% for motor part, actuator part, and blood sac part, respectively. The pump performance assessed in the in vitro Donovan-type mock circulation loop test was acceptable as a BVAD in terms of flow and pressure.


Current status of therapeutic plasma exchange in Korea.
A nationwide survey on the status of plasma exchange in Korea was performed during the 2 year period 2001-02. Data from 496 patients were collected from 15 major hospitals. The most common indication was myasthenia gravis (15.3%), followed by thrombotic thrombocytopenic purpura (14.5%) and hemolytic uremic syndrome (9.7%). Clinical improvement was noted in 70.1% of the 415 cases. Plasma exchange by centrifugation alone accounted for 92.4%. Postcentrifugal filtration was carried out in 5.6% and double filtration in 2.0% of treatments. The most common instruments for the centrifugation were Cobe Spectra (71.3%) and Fenwal CS3000 (15.8%). Filtration was performed by either Kuraray KM8300 or Kuraray KM8800. The overall frequency of complications was 11.1% (293/2647 cases), of which symptomatic hypocalcemia was the most common (2.3%).


Current and Emerging Biomarkers Predicting Bone Metastasis Development.
Bone is one of the preferential sites of distant metastases from malignant tumors, with the highest prevalence observed in breast and prostate cancers. Patients with bone metastases (BMs) may experience skeletal-related events, such as severe bone pain, pathological fractures, spinal cord compression, and hypercalcemia, with negative effects on the quality of life. In the last decades, a deeper understanding of the molecular mechanisms underlying the BM onset has been gained, leading to the development of bone-targeting agents. So far, most of the research has been focused on the pathophysiology and treatment of BM, with only relatively few studies investigating potential predictors of risk for BM development. The ability to select such "high-risk" patients could allow early identification of those most likely to benefit from interventions to prevent or delay BM. This review summarizes several evidences for the potential use of specific biomarkers able to predict early the BM development.


Drug-Resistant Epilepsy: Multiple Hypotheses, Few Answers.
Epilepsy is a common neurological disorder that affects over 70 million people worldwide. Despite the recent introduction of new antiseizure drugs (ASDs), about one-third of patients with epilepsy have seizures refractory to pharmacotherapy. Early identification of patients who will become refractory to ASDs could help direct such patients to appropriate non-pharmacological treatment, but the complexity in the temporal patterns of epilepsy could make such identification difficult. The target hypothesis and transporter hypothesis are the most cited theories trying to explain refractory epilepsy, but neither theory alone fully explains the neurobiological basis of pharmacoresistance. This review summarizes evidence for and against several major theories, including the pharmacokinetic hypothesis, neural network hypothesis, intrinsic severity hypothesis, gene variant hypothesis, target hypothesis, and transporter hypothesis. The discussion is mainly focused on the transporter hypothesis, where clinical and experimental data are discussed on multidrug transporter overexpression, substrate profiles of ASDs, mechanism of transporter upregulation, polymorphisms of transporters, and the use of transporter inhibitors. Finally, future perspectives are presented for the improvement of current hypotheses and the development of treatment strategies as guided by the current understanding of refractory epilepsy.


Activity of minocycline against Acinetobacter calcoaceticus var. anitratus (syn. Herellea vaginicola) and Serratia marcescens.
The activity of minocycline and tetracycline against 23 isolates of Acinetobacter calcoaceticus var. anitratus (syn. Herellea vaginicola) and 178 strains of Serratia marcescens was determined by disk and microdilution methods. The results indicate that minocycline is highly active against this species of Acinetobacter, all but one strain being inhibited by 0.007 mug of the antibiotic per ml. Tetracycline was also active, though to a lesser degree, against A. calcoaceticus. Of the 178 strains of S. marcescens tested, only seven (3.9%) had a minimum inhibitory concentration of 2 mug or less of minocycline per ml. Tetracycline was less active than minocycline against S. marcescens; with 2 mug of tetracycline per ml, only 2 of 152 (1.3%) strains were inhibited. At concentrations of 8 and 16 mug of minocycline per ml, which can be achieved in the urine with the usual doses, 44.9 and 63.5% of S. marcescens strains were inhibited, which implies its possible usefulness for the therapy of urinary tract infection due to this organism.


Psychiatric profile of patients with neurodermatitis.
Fifty patients of neurodermatitis attending the dermatology clinic of a general hospital were screened for psychiatric morbidity. Fifty patients suffering from tinea from the same clinic were taken as a control group (age, sex and socio-economic class matched). A majority of neurodermatitis patients were between 21-30 years of age-group (52%), males (84%), married (90%), literate up to primary (42%) and belonged to socio-economic class V (44%). The common sites of neurodermatitis were legs (28%), arms (16%), scrotum (12%) or multiple sites (36%). The duration of symptoms ranged from 1 month to 2 years (average duration was 6 months). The study group also showed significantly higher occupational and psychosexual problems (p < 0.001). The neurodermatitis patients scored significantly higher on N-2 scale and Hamilton rating scale for depression.


Developing research capacity building for Aboriginal & Torres Strait Islander health workers in health service settings.
Aboriginal and Torres Strait Islander health workers (hereafter called health workers) can play a major role in facilitating culturally appropriate health care delivery and program development through the acquisition of improved skills in the planning, monitoring and evaluation of these programs (RCB). However, many Aboriginal and Torres Strait Islander people and communities remain concerned about research and related activities. Health workers are well placed to assist communities to not only embrace research, but to be active players and promoters of relevant, appropriate and acceptable research. One means of achieving the twin goals of RCB and community acceptance and involvement in research, is through health workers undertaking research of health priority issues and evaluation of activities, such as program delivery, that are of direct relevance to their community's aim of improving or enhancing service delivery. This article outlines the development and content of a community-based RCB framework for health workers. The focus is on the major issues that enhance a proactive service delivery model using culturally appropriate research methods. Development process: The RCB framework described here was developed, over a period of time, through community workshops and consultations aimed at deriving general consensus on the key issues and components of a culturally-appropriate, community-based training process. The framework has subsequently been reviewed by Aboriginal and Torres Strait Islander community representatives from across Australia. The overall aim of the framework is to supplement current (institutionally-based) education and training resources for health workers with community-based research training modules. These modules can be tailored to provide research and evaluation skills relevant to health workers taking a more proactive role in facilitating health and wellbeing programs in their own communities. The use of collaborative consultation and participatory methods are intended to be a two-way education process. Course content: A visual pathway is used that encompasses the impact of health and practice in the community for health workers at a grass-roots level. This enables elements of the RCB process to be divided into a series of connected modules. These are: (i 'assessing' Existing Services; (ii) methods and measures for Identifying Need at various levels; (iii) important issues in Program Development; (iv) how the former contributes to Service Improvement; (v) resultant Outcomes that will impact on community and service provision; and (vi) Evaluation Methods and applying findings to service delivery. Active participation by the Aboriginal and Torres Strait Islander community is fundamental for effective research practices and outcomes. The aim is to provide health workers and community members with a working knowledge of research ethics and methods so that they can assist, monitor and steer the development of culturally appropriate research activities that will lead to provision of the highest quality services 'back' to the community. This RCB framework will enable health workers to be more proactive, self-reliant and self-sufficient within their community and healthcare settings.


Conservation of beneficial microbes between the rhizosphere and the cyanosphere.
Biocrusts are phototroph-driven communities inhabiting arid soil surfaces. Like plants, most photoautotrophs (largely cyanobacteria) in biocrusts are thought to exchange fixed carbon for essential nutrients like nitrogen with cyanosphere bacteria. Here, we aim to compare beneficial interactions in rhizosphere and cyanosphere environments, including finding growth-promoting strains for hosts from both environments. To examine this, we performed a retrospective analysis of 16S rRNA gene sequencing datasets, host-microbe co-culture experiments between biocrust communities/biocrust isolates and a model grass (Brachypodium distachyon) or a dominant biocrust cyanobacterium (Microcoleus vaginatus), and metabolomic analysis. All 18 microbial phyla in the cyanosphere were also present in the rhizosphere, with additional 17 phyla uniquely found in the rhizosphere. The biocrust microbes promoted the growth of the model grass, and three biocrust isolates (Bosea sp._L1B56, Pseudarthrobacter sp._L1D14 and Pseudarthrobacter picheli_L1D33) significantly promoted the growth of both hosts. Moreover, pantothenic acid was produced by Pseudarthrobacter sp._L1D14 when grown on B. distachyon exudates, and supplementation of plant growth medium with this metabolite increased B. distachyon biomass by over 60%. These findings suggest that cyanobacteria and other diverse photoautotrophic hosts can be a source for new plant growth-promoting microbes and metabolites.


Protective effects of TRH and its analogues in chemical and genetic models of seizures.
TRH shows strong influence on neuronal excitability and may participate in the regulation of seizures. We investigated the effect of TRH and its stable analogues on seizures induced by intravenous (i.v.) infusion of pentetrazole in rats. The data showed that i.v. administration of TRH (10 and 20 mg/kg), RGH-2202 (0.1 mg/kg) and Z-p-Glu-His-Pro-NH2 (10 mg/kg) increased threshold for pentetrazole-induced clonic seizures, but did not affect the tonic ones. Another stable analogue of TRH, 1p-Glu-Tyr-Pro-NH2 (0.1-10 mg/kg), had no effect on pentetrazole-induced seizures. In further study, effects of TRH and RGH-2202 were examined in WAG/Rij rats, a genetic model of absence epilepsy. TRH (25 and 50 micrograms i.c.v.) decreased dose-dependently the number and mean duration of spike-wave discharges in cortical EEG of WAG/Rij rats at 90 and 120 min after the peptide administration. On the other hand, RGH-2202 (1 microgram i.c.v.) significantly decreased only the number of spike-wave discharges at 60 min post-injection. These results confirm that TRH and some of its analogues have moderate antiepileptic activity.


Vector Integration Sites Identification for Gene-Trap Screening in Mammalian Haploid Cells.
Forward genetic screens using retroviral (or transposon) gene-trap vectors in a haploid genome revolutionized the investigation of molecular networks in mammals. However, the sequencing data generated by Phenotypic interrogation followed by Tag sequencing (PhiT-seq) were not well characterized. The analysis of human and mouse haploid screens allowed us to describe PhiT-seq data and to define quality control steps. Moreover, we identified several blind spots in both haploid genomes where gene-trap vectors can hardly integrate. Integration of transcriptomic data improved the performance of candidate gene identification. Furthermore, we experimented with various statistical tests to account for biological replicates in PhiT-seq and investigated the effect of normalization methods and other parameters on the performance. Finally, we developed: VISITs, a dedicated pipeline for analyzing PhiT-seq data (https://sourceforge.net/projects/visits/).


Patterns of Antiplatelet Therapy in Patients Who Have Experienced an Acute Coronary Event: A Descriptive Study in UK Primary Care.
Antiplatelet (AP) therapy is well established for the secondary prevention of acute coronary events. However, patients may discontinue treatment, often owing to gastrointestinal (GI) complications, leaving them at elevated risk of recurrent cardiovascular events. This descriptive retrospective study assessed trends in prescription of AP agents and coprescription of gastroprotective therapy, after an acute coronary event. Discontinuation of AP therapy within 2 years of an event and factors predicting discontinuation were investigated. The study was conducted in a UK primary care setting from 2000 to 2008; a total of 27, 351 patients aged 50 to 84 years were included in the analysis. Main outcome measures were exposures to low-dose acetylsalicylic acid (ASA), clopidogrel, and proton pump inhibitors (PPIs). At 90 days after an acute coronary event, 85.9% of patients had been prescribed some form of AP therapy and 33.6% of patients who were issued at least 1 ASA prescription in this period were also issued a PPI prescription. The use of dual antiplatelet therapy (DAT) 90 days after an event increased from 2% in 2000 to over 50% in 2008. An estimated 15.1% of patients on ASA monotherapy and 37.5% on DAT discontinued treatment within 1 year. A bleeding event during follow-up, including upper GI bleeding or hemorrhagic stroke, was the strongest predictor of discontinuation. Although most patients were prescribed AP therapy in the 90 days following an acute coronary event, a substantial proportion discontinued DAT or ASA monotherapy within 1 year. It is essential that physicians consider strategies to reduce the risk of discontinuation of AP therapy.


Large-Scale de novo Oligonucleotide Synthesis for Whole-Genome Synthesis and Data Storage: Challenges and Opportunities.
Over the past decades, remarkable progress on phosphoramidite chemistry-based large-scale de novo oligonucleotide synthesis has been achieved, enabling numerous novel and exciting applications. Among them, de novo genome synthesis and DNA data storage are striking. However, to make these two applications more practical, the synthesis length, speed, cost, and throughput require vast improvements, which is a challenge to be met by the phosphoramidite chemistry. Harnessing the power of enzymes, the recently emerged enzymatic methods provide a competitive route to overcome this challenge. In this review, we first summarize the status of large-scale oligonucleotide synthesis technologies including the basic methodology and large-scale synthesis approaches, with special focus on the emerging enzymatic methods. Afterward, we discuss the opportunities and challenges of large-scale oligonucleotide synthesis on de novo genome synthesis and DNA data storage respectively.


Synthesis of β-C-Glycopyranosyl Aldehydes and 2,6-Anhydro-heptitols.
A convenient route has been developed for the diastereoselective synthesis of β-C-glycopyranosyl aldehydes from D-glucose, D-mannose, and D-galactose. The key step in the synthesis of C-glycosyl aldehydes is the aryl driven reductive dehydration on 1-phenyl-2-(2',3',4',6'-tetra-O-acetyl-β-D-glycopyranosyl)ethanone to afford alkenes, which on oxidation afford the desired compounds in good yield. β-C-Glycopyranosyl aldehydes have been converted to 2,6-anhydro-heptitols in quantitative yields. The 2,6-anhydro-heptitols derived from D-mannose and D-galactose are enantiomeric and are useful linkers for the synthesis of macrocycles/amphiphiles of complementary chirality.


Analysis of intravaginal misoprostol 0.2 mg versus intracervical dinoprostone 0.5 mg doses for labor induction at term pregnancies.
Labor-induction methods are used in about 23% of labors. Most commonly, pharmacological methods are used to pre-induct the labor with dinoprostone - a PGE2 analog, and misoprostol - a PGE1 analog. The aim of this study was to evaluate two pharmacological methods of labor induction with the use of prostaglandins applied via an intravagi-nal insert containing misoprostol at a dose of 0.2 mg and intracervical gel containing dinoprostone at a dose of 0.5 mg. This retrospective study was conducted on a group of 50 adult patients qualified for the pre-induction of labor. Following data were recorded: the time from the drug administration to the beginning of regular contractile function, the time from administration to amniotic fluid rupture, the time from medicament administration to the vaginal labor or caesarean section, the duration of I, II and III stages of labor, the delivery method and in the event of caesarean section - the indications for surgery. In comparison to dinoprostone, the misoprostol application was found to shorten the time from drug administration to amniotic fluid rupture by 14.1 hours, the time to the beginning of the first stage of labor by 11.7 hours and from the drug administration to the delivery by 17.3 hours (p-value < 0.05). The duration of the first stage of labor in the misoprostol group was shorter by 1.2 hours than in dinoprostone group (p-value < 0.05). Application of intravaginal insert with misoprostol at a dose of 0.2 mg appears to be a more effective method of labor induction in comparison to intracervical gel with dinoprostone at a dose of 0.5mg. Thorough analysis of these methods requires further studies.


Glyceraldehyde-3-phosphate Dehydrogenase is a Multifaceted Therapeutic Target.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a glycolytic enzyme whose role in cell metabolism and homeostasis is well defined, while its function in pathologic processes needs further elucidation. Depending on the cell context, GAPDH may bind a number of physiologically important proteins, control their function and correspondingly affect the cell's fate. These interprotein interactions and post-translational modifications of GAPDH mediate its cytotoxic or cytoprotective functions in the manner of a Janus-like molecule. In this review, we discuss the functional features of the enzyme in cellular physiology and its possible involvement in human pathologies. In the last part of the article, we describe drugs that can be employed to modulate this enzyme's function in some pathologic states.


Sensitive Detection of MMP9 Enzymatic Activities in Single Cell-Encapsulated Microdroplets as an Assay of Cancer Cell Invasiveness.
Matrix metalloproteinases (MMPs) are typically up-regulated in cancer cells, and play a critical role in assisting metastasis by the breakdown of the extracellular matrix. Here we report an effective strategy for cell invasiveness assay by integrating MMP9 functional activity analysis with single cell-encapsulated microdroplets. A flow focusing capillary microfluidic device has been assembled using "off-the-shelf" fluidic components for high-throughput generation of microdroplets. Tumor cells, MMP9 specific peptides, and other cofactors can be loaded into the device and encapsulated into individual droplets as dynamic microreactors for proteolytic cleavage of the substrate. This design allows for rapid and robust detection of MMP9 enzymatic activities by fluorescent signals in a few minutes. It represents the first demonstration of quantifying MMP9 enzymatic activities at the single cell level with a high throughput performance. This new technique promises functional evaluation of cancer cell invasiveness for important diagnostic or prognostic applications.


Management of Severe Equinovalgus in Patients With Cerebral Palsy by Naviculectomy in Combination With Midfoot Arthrodesis.
Equinovalgus deformity is the second most common deformity in cerebral palsy and may be flexible or rigid. Several operative methods from joint sparing to arthrodesis have been described with varying success rates. The aim of this study was to investigate the effectiveness of naviculectomy in combination with midfoot arthrodesis (talo-cuneiform and calcaneocuboid arthrodesis) in the correction of a rigid equinovalgus foot deformity in cerebral palsy. Forty-eight rigid equinovalgus feet were operated upon in 30 patients from 2008 to 2013. Of these, 44 feet in 26 patients with cerebral palsy (Gross Motor Function Classification System III, IV, or V) with follow-up of more than 2 years were included in the study. The mean age at surgery was 18.1 years. The outcomes were measured objectively using radiographic angles and subjectively using 5 questions to be answered by the caregiver. The feet were then graded into excellent, good, fair, and poor. The mean follow-up was 5.0 ± 1.7 years. Excellent to good results were obtained in 81% of the feet. Both objective and subjective outcomes improved significantly postoperatively ( P < .001). Three feet in 2 patients were graded as poor and underwent a revision operation for pain and recurrence. Naviculectomy in combination with midfoot arthrodesis enabled a good 3-dimensional correction of the forefoot. However, the procedure did not necessarily correct the fixed subtalar joint deformity. Several additional bony and soft-tissue procedures were necessary to achieve a complete correction in these difficult feet. Level IV, retrospective case series.


An ex vivo perfusion system emulating in vivo conditions in noncirrhotic and cirrhotic human liver.
Various models are used for investigating human liver diseases and testing new drugs. However, data generated in such models have only limited relevance for in vivo conditions in humans. We present here an ex vivo perfusion system using human liver samples that enables the characterization of parameters in a functionally intact tissue context. Resected samples of noncirrhotic liver (NC; n = 10) and cirrhotic liver (CL; n = 12) were perfused for 6-h periods. General and liver-specific parameters (glucose, lactate, oxygen, albumin, urea, and bile acids), liver enzymes (aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, glutamate dehydrogenase, and γ-glutamyl transferase), overall (M65) and apoptotic (M30) cell-death markers, and indicators of phase-I/phase-II biotransformations were analyzed. The measurement readings closely resembled (patho)physiological characteristics in patients with NC and CL. Mean courses of glucose levels reflected the CLs' reduced glycogen storage capability. Furthermore, CL samples exhibited significantly stronger increases in lactate, bile acids, and the M30/M65 ratio than NC specimens. Likewise, NC samples exhibited more rapid phase-I transformations of phenacetin, midazolam, and diclofenac and phase-I to phase-II turnover rates of the respective intermediates than CL tissue. Collectively, these findings reveal the better hepatic functionality in NC. Perfusion of human liver tissue with this system emulates in vivo conditions and clearly discriminates between noncirrhotic and cirrhotic tissue. This highly reliable device for investigating basic hepatic functionality and testing safety/toxicity, pharmacokinetics/pharmacodynamics and efficacies of novel therapeutic modalities promises to generate superior data compared with those obtained via existing economic perfusion systems.


Heterologous expression and purification of NisA, the precursor peptide of lantibiotic nisin from Lactococcus lactis.
The lantibiotic nisin is a ribosomally synthesised and post-translationally modified antimicrobial peptide produced by strains of Lactococcus lactis, and used as safe and natural preservative in food industry. The nisA structural gene encodes ribosomally synthesised and biologically inactive a 57 amino acid precursor peptide (NisA) which undergoes several post-translational modifications. In this study, we report the expression of precursor nisin as a His6-tagged peptide in Escherichia coli and its purification using a nickel affinity column. The technique of spliced-overlap extension PCR was used to amplify the nisA gene and the T7 promoter region of pET-15b vector. This approach was used to introduce six histidine residues at the C-terminus of prenisin. The identity of the expressed peptide was confirmed by N-terminal sequencing. The expressed His-tagged prenisin was purified under denaturing conditions, and named as prenisin-His6. The purified prenisin-His6 was analyzed by SDS-PAGE, Western blotting and mass spectroscopy. These results showed that the nisin precursor peptide can be successfully produced using an E. coli expression system.


Benchmarking the DFT+U method for thermochemical calculations of uranium molecular compounds and solids.
Ability to perform a feasible and reliable computation of thermochemical properties of chemically complex actinide-bearing materials would be of great importance for nuclear engineering. Unfortunately, density functional theory (DFT), which on many instances is the only affordable ab initio method, often fails for actinides. Among various shortcomings, it leads to the wrong estimate of enthalpies of reactions between actinide-bearing compounds, putting the applicability of the DFT approach to the modeling of thermochemical properties of actinide-bearing materials into question. Here we test the performance of DFT+U method--a computationally affordable extension of DFT that explicitly accounts for the correlations between f-electrons - for prediction of the thermochemical properties of simple uranium-bearing molecular compounds and solids. We demonstrate that the DFT+U approach significantly improves the description of reaction enthalpies for the uranium-bearing gas-phase molecular compounds and solids and the deviations from the experimental values are comparable to those obtained with much more computationally demanding methods. Good results are obtained with the Hubbard U parameter values derived using the linear response method of Cococcioni and de Gironcoli. We found that the value of Coulomb on-site repulsion, represented by the Hubbard U parameter, strongly depends on the oxidation state of uranium atom. Last, but not least, we demonstrate that the thermochemistry data can be successfully used to estimate the value of the Hubbard U parameter needed for DFT+U calculations.


Results of treatment for craniopharyngioma.
A review of the results of treatment for 155 cases of craniopharyngioma during the past 47 years by the palliative operation was made. Steroid and operative microscope reduced operative mortality and gave more chance of total extirpation, but survival rate was almost unchanged. It is noteworthy that 3 cases who underwent palliative operation more than 30 years ago are still alive and leading useful lives. The quality of survival was somewhat less favorable in pediatric than adult cases. Morbidity frequently encountered in pediatric cases was growth retardaton. Craniopharyngioma is a benign tumor pathohistologically, but it should be considered as malignant because of the region it occurs in. Therefore, no forceful attempt to total extirpation should be made and the operations should, as a rule, be palliative.


Ciprofibrate, clofibric acid and respective glycinate derivatives. Effects of a four-week treatment on male lean and obese Zucker rats.
Fibrates are widely prescribed in hyperlpidemic patients to prevent atherosclerosis. Their therapeutic use, however, can be associated with adverse effects like gastrointestinal disorders, myalgia, myositis and hepatotoxicity. In rodents large doses can even cause hepatocellular carcinoma. Additionally, interactions with the biotransformation of other compounds at the cytochrome P450 (CYP) system have been observed. Thus, the discovery of new substances or derivatives with less side effects is of great interest. In the present study the influence of a four-week daily oral administration of 2 mg/kg body weight ciprofibrate (CAS 52214-84-3) or of 100 mg/kg body weight clofibric acid (CAS 882-09-7) was compared to that of the respective doses of their newly synthesized glycine conjugates in adult male lean and obese Zucker rats. Although obese rats displayed distinctly higher serum lipid concentrations, after fibrate treatment values were significantly lowered in lean animals only. Livers of obese rats were significantly enlarged, histologically showing a fine-droplet like fatty degeneration and an increase in glycogen content, but no signs of inflammation. After fibrate administration histologically a hypertrophy, an eosinophilia, a reduced glycogen content and also hepatocyteapoptosis were observed. Livers of obese rats displayed higher CYP1A1 andCYP2E1 expression, but lower immunostaining for CYP2B1 and CYP3A2. No differences between the two groups of rats were seen with respect to CYP4A1 expression. Due to fibrate treatment especially CYP2E1 and CYP4A1, but also CYP1A1, 2B1 and 3A2 were induced. Resulting CYP mediated monooxygenase activities were also elevated in most cases. In general, effects of clofibric acid and clofibric acid glycinate (CAS 4896-55-3) were less distinct than those of ciprofibrate and its glycinate (CAS 640772-36-7). With no parameterinvestigated major differences were seen between the parent fibrates and their glycine conjugates. Thus, the present investigations revealed no noticeable advantages of the glycinates over ciprofibrate or clofibric acid.


Effect of 60Co-irradiation on the development and immunogenicity of Plasmodium berghei sporozoites in Anopheles stephensi mosquitoes.
Protection conferred to mice by Plasmodium berghei sporozoites increased significantly when the time interval between 60Co-irradiation of the infected mosquitoes and harvest of sporozoites increased. One thousand sporozoites conferred no protection against challenge if harvested on the day of irradiation, but protected 60% of recipient mice when harvested 28 days postirradiation. When the time between feeding of mosquitoes and irradiation was varied, sporozoites from mosquitoes irradiated 3 days after feeding were infective for mice. Sporozoites from mosquitoes irradiated on day 10 postfeeding were not infective, but were immunogenic. In all experiments a decline occurred in the number of recoverable sporozoites over a 28-day period postirradiation to less than 10% of the yield on the day of irradiation.


Age differences in, and consequences of, multiple- and variable-strategy use on a multitrial sort-recall task.
Second-, 3rd-, and 4th-grade children were given 5 sort-recall trials with different sets of categorizable items. The authors assessed multiple-strategy use on each trial and related intertrial changes in strategy use to levels of recall. Multiple-strategy use increased with age but was observed at all grades. Fourth graders who used more strategies had higher recall, with this pattern occurring only on later trials for 2nd and 3rd graders. Children of all ages showed substantial intertrial variability in using multiple strategies. Stable-strategy use (few strategy changes across trials) was related to high levels of recall, both for individual and group data, and was associated with the use of different numbers of strategies at different ages. Results were interpreted in light of new research and theory postulating that multiple- and variable-strategy use is the rule rather than the exception in development and that strategies do not always facilitate task performance (utilization deficiencies).