The Application of Microwaves, Ultrasounds, and Their Combination in the Synthesis of Nitrogen-Containing Bicyclic Heterocycles.
The use of alternative energy sources, such as microwaves (MW) or ultrasounds (US), and their mutual cross-combination have been widely described in the literature in the development of new synthetic methodologies in organic and medicinal chemistry. In this review, our attention is focused on representative examples, reported in the literature in the year range 2013-2023 of selected N-containing bicyclic heterocycles, with the aim to highlight the advantages of microwave- and ultrasound-assisted organic synthesis.


Downregulation of NGAL is Required for the Inhibition of Proliferation and the Promotion of Apoptosis of Human Gastric Cancer MGC-803 Cells.
Gastric cancer is considered as a common malignancy with a poor prognosis as well as unsatisfactory treatment. Neutrophil gelatinase-associated lipocalin (NGAL) has been reported to affect multiple aspects of human tumor, including gastric cancer. This study aims to explore the effects of NGAL gene silencing on the proliferation as well as apoptosis of human gastric cancer MGC-803 cells. This study included 87 patients with gastric cancer. MGC-803 cells were collected and mainly treated with siRNA against NGAL and recombinant NGAL plasmid. The expression of NGAL mRNA and the expressions of NGAL protein and apoptosis-related proteins were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis, respectively. Cell cycle and apoptosis were tested by flow cytometry, and cell proliferation was detected by water soluble tetrazolium-1 (WST-1) assay. The effect of NGAL gene silencing on tumorigenicity of MGC-803 cells in vivo was detected through establishment of xenograft in nude mice. NGAL was highly expressed in gastric cancer tissues. The protein and mRNA expressions of NGAL gene in MGC-803 cells treated with NGAL-siRNA were obviously reduced, and the amount of cells in G0/G1 phase was increased. Moreover, MGC-803 cells treated with NGAL-siRNA exhibited inhibited proliferation, enhanced apoptosis, decreased expressions of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) as well as B-cell lymphoma-2 (Bcl-2) and increased expressions of cysteine-aspartic acid specific protease-9 (caspase-9) and Bcl2-associated X (Bax), as well as repressed tumorigenicity in vivo. NGAL gene silencing inhibits proliferation and promotes apoptosis of MGC-803 cells, which can provide a novel theory for treatment of gastric cancer.


Conversion of fatty aldehyde dimethyl acetals to the corresponding alk-1-enyl methyl ethers (substituted vinyl ethers) during gas-liquid chromatography.
The behavior of palmitaldehyde and linolealdehyde and of their dimethyl acetals during gas-liquid chromatography on beta-cyclodextrin acetate (beta-CDX acetate) and ethylene glycol succinate polyester-phosphoric acid (EGSP) columns was studied. The aldehydes were well separated from their dimethyl acetals on the beta-CDX acetate column. However, on the EGSP column the retention times of palmitaldehyde and its dimethyl acetal were identical; a mixture of linolealdehyde and its dimethyl acetal gave a split peak. The aldehydes were recovered unchanged in 80-85% yield by preparative GLC from both columns, but the dimethyl acetals were quantitatively converted to the corresponding alk-1-enyl methyl ethers. The structure of these compounds was elucidated by infrared spectroscopy, mass spectrometry, and chemical means. Upon hydrolysis at low temperatures with 100% H(2)SO(4) they yielded the corresponding aldehydes, which were identified as 2,4-dinitrophenylhydrazones.


Sympathetic activation in resistant hypertension: theory and therapy.
Resistant hypertension defined as requiring 3 or more complementary antihypertensive drugs at maximally tolerated doses accounts for approximately 3% to 4% of all cases of hypertension. Its increased incidence over the past decade is related to the increase in obesity in the Western world. There are a number of dietary factors that affect sympathetic tone including sodium intake apart from increased body mass. This article discusses the mechanisms of sympathetic stimulation and activation in the context of animal models and human studies. In addition, there is a review of clinical trials with and without device therapy that summarizes the clinical findings. Effective management should be based on pathophysiologic principles and a focus on blood pressure reduction to levels well below 150/90 mm Hg because outcome trial evidence and Food and Drug Administration guidance supports this construct. The key to success of device-based therapy depends on identifying the cohort with true resistant hypertension that can benefit from therapies that are adjuncts to pharmacotherapy. Physicians need to concentrate on educating the patient on lifestyle modifications and themselves on use of proper combinations of antihypertensive medications. If this approach fails to result in a safe level of blood pressure then the patient should be referred to a board-certified clinical hypertension specialist.


A Case of Febrile Polyphagia With an Underlying Paraneoplastic Limbic Encephalitis.
A 57-year-old female with a history of malignant mixed Müllerian tumors of the uterus and ovaries developed a fever of unknown origin and neuropsychiatric symptoms. Her EEG showed slow activity in the left temporal region, and brain magnetic resonance imaging revealed limbic encephalitis, leading to the diagnosis of classic paraneoplastic limbic encephalitis (PLE). During our investigation into the underlying cause of the patient's condition, we conducted a PET-CT scan, which revealed the presence of several hypermetabolic lymph nodes. One of these lymph nodes underwent a biopsy, and the results confirmed the presence of metastatic cells, indicating the likelihood of carcinoma, most probably adenocarcinoma of the gynecological tract. PLE should be considered as one of the differential diagnoses in patients with a history of cancer and acute-to-subacute neuronal and psychiatric dysfunction.


[Expression of HIF-1a in acute limb ischemia of rats].
To explore the expression of hypoxia-inducible factor 1 alpha (HIF-1a) in rat lower limb skeletal muscle with acute ischemia. Acute lower limb ischemia was established in 25 male or female SD rats of 200-250 g by ligating left iliac artery with microsurgical techniques. Gastrocnemius of lower limb was harvest at the time points of sham (0h), 4, 8, 12 and 24 h respectively. Immunohistochemistry, Western blot and real-time polymerase chain reaction (PCR) were used to detect the expression of HIF-1a in lower limb gastrocnemius. There was a low expression of HIF-1a in normal gastrocnemius, but it was highly expressed in lower limb ischemia. With the elongation of ischemia time, its expression increased (P < 0.05), peaked at 4 h and 8 h (P < 0.05) and then declined gradually. With a characteristic trend during acute limb ischemia, HIF-1a may play an important role in the adaptive response to acute limb ischemia.


Framework for defining pesticide maximum residue levels in feed: applications to cattle and sheep.
Pesticide residues in animal feed can endanger animal health and compromise the safety of livestock products for human consumption. Even though policymakers such as the European Union and the World Health Organization have established maximum residue levels (MRLs) for pesticides in both human food and animal feed, there is no systematic management of pesticides in animal feed that considers the entire supply chain. In response, we propose a framework for defining consistent MRLs for pesticides in animal feed that assesses the impact of defined MRLs on upstream (e.g., MRLs in feed crops) and downstream (e.g., MRLs in livestock products) sectors of the livestock-product supply chain. The MRLs determined for the selected pesticides in the feed of cattle and sheep as case study animals indicate that lipophilic pesticides tend to have lower MRLs than hydrophilic pesticides, primarily due to the relatively high toxicity and biotransfer factors of lipophilic pesticides. In addition, we observe that, primarily for lipophilic pesticides, upstream and downstream regulations are not aligned in terms of defining MRLs in feed using current MRLs in crops with relevance to feed and foods of animal origin. Some of the current pesticide regulations in the livestock-product supply chain need to be re-evaluated to ensure that MRLs in the upstream sector (i.e., crops) do not result in unacceptable residues in the downstream sector (i.e., MRLs in livestock products affecting animal and human health). Finally, we provide recommendations for optimizing the derivation of MRLs in feed, including the evaluation of residue fate during feed and food manufacturing processes. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Nonlinear Optical Modulation Characteristics of MXene Cr2C for 2 μm Pulsed Lasers.
MXene materials have shown numerous useful mechanical and electronic properties, and have been found to possess nice potential in the field of optical modulation. Here, we fabricated a MXene Cr2C saturable absorber by the liquid-phase exfoliation method, and systemically analyzed the surface morphology and nonlinear properties of the Cr2C sample. Applying the Cr2C saturable absorber as a Q-switch in a thulium-doped yttrium aluminum perovskite (Tm: YAP) laser, the shortest single pulse was obtained with a width of 602 ns under an absorbed pump power of 3.3 W at a repetition rate of 55 kHz with a T = 1% output coupler. The maximum output power was obtained with a T = 5% output coupler at a repetition rate of 58 kHz. The obtained maximum pulse energy and peak power were 3.96 μJ and 4.36 W, separately, which reveal that the MXene Cr2C can be applied as a promising modulation material in the near-infrared pulsed lasers.


Cellular energy supply for promoting vascular remodeling of small-diameter vascular grafts: a preliminary study of a new strategy for vascular graft development.
Rapid endothelialization is extremely essential for the success of small-diameter tissue-engineered vascular graft (TEVG) (<6 mm) transplantation. However, severe inflammation in situ often causes cellular energy decline of endothelial cells. The cellular energy supply involved in vascular graft therapy remains unclear, and whether promoting energy supply would be helpful in the regeneration of vascular grafts needs to be established. In our work, we generated an AMPK activator (5-aminoimidazole-4-carboxamide ribonucleotide, AICAR) immobilized vascular graft. AICAR-modified vascular grafts were successfully generated by the co-electrospinning technique. In vitro results indicated that AICAR could upregulate energy supply in endothelial cells and reprogram macrophages (MΦ) to assume an anti-inflammatory phenotype. Furthermore, endothelial cells (ECs) co-cultured with AICAR achieved higher survival rates, better migration, and angiogenic capacity than the controls. Concurrently, a rabbit carotid artery transplantation model was used to investigate AICAR-modified vascular grafts at different time points. The results showed that AICAR-modified vascular grafts had higher patency rates (92.9% and 85.7% at 6 and 12 weeks, respectively) than those of the untreated group (11.1% and 0%). In conclusion, AICAR strengthened the cellular energy state and attenuated the adverse effects of inflammation. AICAR-modified vascular grafts achieved better vascular remodeling. This study provides a new perspective on promoting the regeneration of small-diameter vascular grafts.


Considerations for dosing immunoglobulin in obese patients.
Obesity is a very common condition; however, the effect of excess body weight on the appropriate dose of immunoglobulin has not been defined empirically. The proposed pharmacokinetic differences between lean and obese patients and the opportunity to reduce costs has led to the proposition that obese patients should receive proportionally lower doses of immunoglobulin once a certain threshold is reached. Here the theoretical factors which could affect dosing in obese patients are considered alongside the available empirical evidence. The available evidence indicates that obesity may affect the pharmacokinetics of immunoglobulin; however, the effect is likely to be too small to have a clinically important effect on dosing. Wide interpatient individuality and highly variable clinical need mean that obesity should not play a major factor in dosing considerations. However, patients who are obese are more likely to have multiple cardiovascular risk factors and their weight indicates a large dose. This puts these patients at a higher risk of adverse reactions, and therefore caution is advised.


Effects of BMAL1 Manipulation on the Brain's Master Circadian Clock and Behavior.
Bmal1 is the only single circadian clock gene that is essential for rhythmic gene expression in the mammalian circadian timing system. Genetic approaches targeting Bmal1 expression have been used to further assess its role in the circadian clock and to test for behavioral effects of clock disruption. In particular, disruptions in circadian clock function have been implicated in human mood disorders, and clock gene manipulation in mice may provide valuable models for studying depression-like behavior. In this review, we explore various approaches to manipulating Bmal1 in mouse models and review their effects on the brain's master circadian pacemaker, on circadian rhythmicity in other brain regions, and on circadian and mood-related behavior.


FACTORS INFLUENCING SIZE-SPECIFIC DOSE ESTIMATES OF SELECTED COMPUTED TOMOGRAPHY PROTOCOLS AT TWO CLINICAL PRACTICES IN SOUTH AFRICA.
The study aimed to determine the factors that impact the size-specific dose estimate (SSDE) for computed tomography (CT) examinations of the chest-abdomen-pelvis and abdomen-pelvis protocols in two clinical radiology practices and evaluate the image quality of these protocols. Imaging parameters, protocols, dose metrics from the CT units and size-related parameters to calculate the SSDE were documented. The image quality of the CT images was assessed using an image subtraction algorithm. The SSDE increased as the volumetric CT dose index (CTDIvol), and the patient's body mass index increased, respectively. Significant differences (p < 0.001) occurred between the two hospitals regarding image quality. However, these differences were not indicative of differences in the diagnostic performances for task-based imaging protocols. Different clinical protocols should be reviewed to optimise dose. The inclusion of the pre-monitoring sequence, age of the machine and the scan requisition parameters impacted the SSDEs. Image quality should be assessed to evaluate the consistency of image quality between protocols applied by different CT units when assessing SSDEs.


An amygdalar neural ensemble that encodes the unpleasantness of pain.
Pain is an unpleasant experience. How the brain's affective neural circuits attribute this aversive quality to nociceptive information remains unknown. By means of time-lapse in vivo calcium imaging and neural activity manipulation in freely behaving mice encountering noxious stimuli, we identified a distinct neural ensemble in the basolateral amygdala that encodes the negative affective valence of pain. Silencing this nociceptive ensemble alleviated pain affective-motivational behaviors without altering the detection of noxious stimuli, withdrawal reflexes, anxiety, or reward. Following peripheral nerve injury, innocuous stimuli activated this nociceptive ensemble to drive dysfunctional perceptual changes associated with neuropathic pain, including pain aversion to light touch (allodynia). These results identify the amygdalar representations of noxious stimuli that are functionally required for the negative affective qualities of acute and chronic pain perception.


The mechanism of catheter kickback in the final stage of coil embolization for aneurysms: the straightening phenomenon.
The catheter kickback phenomenon often occurs in the last stage of coil packing for cerebral aneurysms. This behavior is considered the result of either a lack of space in the sac or a stiff detaching zone. In order to clarify its mechanism, focused stretch-resistance (SR) coil simulation models were tested. Various commercially available SR coils were inserted into a cylinder or an aneurismal sac made from a silicone tube with a smaller than prescribed diameter. A magnified SR coil model (straight type) of fishing line was created for simulation. Numerical analysis for the changes in coil behavior were verified. All SR coils showed hardening and straightening at the last few millimeters, resulting in catheter kickback. In a magnified coil experiment, straightening was also realized when folding into a narrow cylinder. The SR line coursed in the canal of the first loops and shifted to the outside in the middle portion. Gaps among first coil pitches were enlarged on after insertion into the narrower space. Shortage of the SR line was calculated to reach a maximum of 32%. The straightening phenomenon is due to SR line shortening and subsequent condensation of pitches of the first loops at the coil end. Coil tail flexibility was lost, and the coil behaved as a stiff wire. Straightening is an important factor in the kickback phenomenon. Shorter final SR coils should be selected, and coil designs should be improved.


Opioid antagonist effects on self-injury in adults with mental retardation: response form and location as determinants of medication effects.
The opioid antagonist naltrexone was administered to 8 adults with severe or profound mental retardation and extensive histories of self-injurious behavior. Five-minute behavioral samples were observed randomly out of every hour from 8 a.m. through 3 p.m., Monday through Friday, for four 2-week phases (baseline, placebo, 50 mg, and 100 mg). During naltrexone administration, there were fewer days with frequent head-banging and self-biting, whereas there were more days on which blows to the head or self-biting were infrequent. Self-injurious participants slept 1.38 hours less per night during baseline, which was unaffected by naltrexone.


The Effects of Age and Reproduction on the Lipidome of Caenorhabditis elegans.
Aging is a complex life process, and a unified view is that metabolism plays key roles in all biological processes. Here, we determined the lipidomic profile of Caenorhabditis elegans (C. elegans) using ultraperformance liquid chromatography high-resolution mass spectrometry (UPLC-HRMS). Using a nontargeted approach, we detected approximately 3000 species. Analysis of the lipid metabolic profiles at young adult and ten-day-old ages among wild-type N2, glp-1 defective mutant, and double mutant daf-16;glp-1 uncovered significant age-related differences in the total amount of phosphatidylcholines (PC), sphingomyelins (SM), ceramides (Cer), diglycerides (DG), and triglycerides (TG). In addition, the age-associated lipid profiles were characterized by ratio of polyunsaturated (PUFA) over monounsaturated (MUFA) lipid species. Lipid metabolism modulation plays an important role in reproduction-regulated aging; to identify the variations of lipid metabolites during germ line loss-induced longevity, we investigated the lipidomic profiles of long-lived glp-1/notch receptor mutants, which have reproductive deficiency when grown at nonpermissive temperature. The results showed that there was some age-related lipid variation, including TG 40:2, TG 40:1, and TG 41:1, which contributed to the long-life phenotype. The longevity of glp-1 mutant was daf-16-dependent; the lipidome analysis of daf-16;glp-1 double mutant revealed that the changes of some metabolites in the glp-1 mutant were daf-16-dependent, while other metabolites displayed more complex epistatic patterns. We first conducted a comprehensive lipidome analysis to provide novel insights into the relationships between longevity and lipid metabolism regulated by germ line signals in C. elegans.


The International Classification of Functioning, Disability and Health Core Set for deafblindness. Part I: a systematic review of outcome measures.
The International Classification of Functioning, Disability, and Health (ICF), developed by the World Health Organization, is a classification framework that focuses on the health and functioning of people with disabilities. As part of an ICF Core Set development, four studies need to be conducted, one of which is a systematic review. This study presents part 1 of the systematic review that aims to describe the outcome measures identified in the literature related to functioning in individuals with deafblindness. The research team screened articles from eight scientific databases, three journals, and Google Scholar (March 2011 to September 2022). Articles were included if they studied individuals with deafblindness aged 18 and older. Studies that examined genetics or laboratory experiments involving animals were excluded. Data were extracted into a logbook with key descriptors such as study location and design, age of study population, and instruments/outcome measures used, which were further categorized into one of the following types: 1) standardized; 2) patient-reported measures, standardized (PT-S); 3) patient-reported measures, not standardized (PT-not S); 4) health professional, reported measures, standardized (HP-S); 5) Technical measures; 6) other measures (parent-reported standardized and laboratory measures). The review included 147 studies, of which most were conducted in Europe (47.6%) and North America (27.9%). Of the 314 identified outcome measures, 57 were Standardized, 59 were Patient Reported-Standardized (PT-S), 178 were patient reported non-standardized (PT-Not S) variables, 11 were health professional reported, standardized, five were technical, and four were classified as other measures. Most instruments measured functioning in daily activities and the mental health of individuals with deafblindness. Three deafblind-specific instruments were identified in this study, highlighting the need for more deafblind-specific instruments to be developed and utilized in research.


Antioxidant rich Morus alba leaf extract induces apoptosis in human colon and breast cancer cells by the downregulation of nitric oxide produced by inducible nitric oxide synthase.
Morus species had been used widely in the traditional medicines for various diseases. In this study we report the in vitro antiproliferative activity of the methanol extract of Morus alba. The extract is capable of inducing cytotoxicity in human colon cancer (HCT-15) cells (IC(50) = 13.8 μg/ml) and breast cancer (MCF-7) cells (IC(50) = 9.2 μg/ml), resulted in significant morphological changes of the cells, fragmentation of DNA, and caspase-3 activation- characteristics of apoptosis. It downregulated the amount of nitric oxide (NO) produced as a result of inducible nitric oxide (iNOS) activation. The HPLC analysis of the extract showed epicatechin (20%), myricetin (10%), quercetin hydrate (12%), luteolin (12%), and kaempferol (6%) as the major active components and ascorbic acid, gallic acid, pelargonidine, and p-coumaric acid as the minor components. To the best of our knowledge, this is the first report showing the downregulation of iNOS and induction of apoptosis by M. alba extract.


Development and evaluation of a real-time RT-PCR assay on the LightCycler for the rapid detection of enterovirus in cerebrospinal fluid specimens.
Detection of enteroviral nucleic acid in cerebrospinal fluid (CSF) specimens has been demonstrated to improve the management of patients with aseptic meningitis. To develop on the LightCycler (LC) instrument a real-time RT-PCR assay based on TaqMan technology for the detection of enteroviruses (EV) in cerebrospinal fluid (CSF) specimens. After evaluation of the analytical performances, seventy-four CSF samples collected prospectively from patients who have been suspected for a clinical diagnosis of meningitis were evaluated by two LC real-time RT-PCR assays and one conventional RT-PCR assay. Our assay detected all 30 different EV species tested, whereas no reactivity was observed with other neurotropic viruses. The analytical sensitivity of both LC RT-PCR real-time assays was 1 TCID50 for LC one-step and two-step RT-PCR assays. Results for LC one-step and LC two-step RT-PCR were compared to results of the conventional RT-PCR: of the 74 CSF specimens tested, 11 were positive and 56 were negative by all methods. Four other specimens were positive for EV by at least two of the methods (including the LC two-step RT-PCR and the conventional RT-PCR), two other CSF specimens were positive by the LC two-step RT-PCR assay only, and another one CSF specimen was positive by the LC one-step RT-PCR assay only. No CSF specimens were negative by the LC two-step RT-PCR assay and positive by the conventional RT-PCR assay. The sensitivity, specificity, positive and negative predictive values of both LC RT-PCR assays by using conventional RT-PCR as the "gold standard" were, respectively, 73.3, 98.3, 91.7, 93.5% for the LC one-step RT-PCR and 100, 96.6, 88.2, 100% for the LC two-step RT-PCR. There was substantial agreement between the three assays (k=0.80). The LC two-step RT-PCR assay is a rapid, sensitive and reliable method which can be routinely performed with CSF samples for diagnosis of EV infection and is an important improvement for optimal patient management.


Analytical procedure for the simultaneous voltammetric determination of trace metals in food and environmental matrices. Critical comparison with atomic absorption spectroscopic measurements.
An analytical procedure fit for the simultaneous determination of copper (II), chromium(VI), thallium(I), lead(II), tin(II), antimony(III), and zinc(II) by square wave anodic stripping voltammetry (SWASV) in three interdependent environmental matrices involved in foods and food chain as meals, cereal plants and soils is described. The digestion of each matrix was carried out using a concentrated HCl-HNO3-H2SO4 (meals and cereal plants) and HCl-HNO3 (soils) acidic attack mixtures. 0.1 mol/L dibasic ammonium citrate pH 8.5 was employed as the supporting electrolyte. The voltammetric measurements were carried out using, as working electrode, a stationary hanging mercury drop electrode (HMDE) and a platinum electrode and an Ag/AgCl/KClsat electrode as auxiliary and reference electrodes, respectively. The analytical procedure was verified by the analyses of the standard reference materials: Wholemeal BCR-CRM 189, Tomato Leaves NIST-SRM 1573a and Montana Soil Moderately Elevated Traces NIST-SRM 2711. For all the elements in the certified matrix, the precision as repeatability, expressed as relative standard deviation (Sr %) was lower than 5%. The accuracy, expressed as percentage relative error (e %) was of the order of 3-7%, while the detection limits were in the range 0.015-0.103 microg/g. Once set up on the standard reference materials, the analytical procedure was transferred and applied to commercial meal samples, cereal plants and soils samples drawn in sites devoted to agricultural practice. A critical comparison with spectroscopic measurements is also discussed.


Bupropion attenuates nicotine abstinence syndrome in the rat.
Bupropion reduces discomfort and craving associated with smoking cessation. This study determined whether a rat model of nicotine dependence could detect such nicotine abstinence-alleviating effects. Experiments determined whether the abstinence-alleviating effects of bupropion were detectable by (1) behavioral abstinence signs precipitated by the nicotinic antagonist mecamylamine, (2) place aversion conditioned to mecamylamine-precipitated nicotine abstinence, and (3) spontaneous behavioral abstinence signs after abrupt nicotine withdrawal. In experiments 1 and 2, nicotine-dependent rats were coinfused for 7 days with 3.15 mg/kg/day nicotine and 20 mg/kg/day bupropion or with nicotine alone. They were then challenged with 1 mg/kg mecamylamine and observed for behavioral abstinence signs (experiment 1) or place aversion conditioned to precipitated abstinence (experiment 2). In experiment 3, rats were nicotine-infused for 7 days as above. A day after termination of nicotine infusion, rats were observed for spontaneous nicotine abstinence signs before and after injection with saline or bupropion. In experiment 1, rats coinfused with nicotine and bupropion had significantly fewer mecamylamine-precipitated abstinence signs than rats infused with nicotine alone but similar numbers to rats infused with saline alone. In experiment 2, bupropion pretreatment significantly reduced the aversiveness of mecamylamine-precipitated nicotine abstinence. In experiment 3, a single bupropion injection dose-dependently alleviated spontaneous nicotine abstinence syndrome. These results suggest that these rat models of nicotine dependence and abstinence syndrome may be useful in detecting nicotine abstinence-alleviating effects of potential medications for smoking cessation. The effects of acute bupropion administration raise interesting questions regarding bupropion's mechanism of action.


High-mass matrix-assisted laser desorption ionization-mass spectrometry of integral membrane proteins and their complexes.
Analyzing purified membrane proteins and membrane protein complexes by mass spectrometry has been notoriously challenging and required highly specialized buffer conditions, sample preparation methods, and apparatus. Here we show that a standard matrix-assisted laser desorption/ionization (MALDI) protocol, if used in combination with a high-mass detector, allows straightforward mass spectrometric measurements of integral membrane proteins and their complexes, directly following purification in detergent solution. Molecular weights can be determined precisely (mass error ≤ 0.1%) such that high-mass MALDI-MS was able to identify the site for N-linked glycosylation of the eukaryotic multidrug ABC transporter Cdr1p without special purification steps, which is impossible by any other current approach. After chemical cross-linking with glutaraldehyde in the presence of detergent micelles, the subunit stoichiometries of a series of integral membrane protein complexes, including the homomeric PglK and the heteromeric BtuCD as well as BtuCDF, were unambiguously resolved. This thus adds a valuable tool for biophysical characterization of integral membrane proteins.


Quantitative Characterization of the Binding and Unbinding of Millimolar Drug Fragments with Molecular Dynamics Simulations.
A quantitative characterization of the binding properties of drug fragments to a target protein is an important component of a fragment-based drug discovery program. Fragments typically have a weak binding affinity, however, making it challenging to experimentally characterize key binding properties, including binding sites, poses, and affinities. Direct simulation of the binding equilibrium by molecular dynamics (MD) simulations can provide a computational route to characterize fragment binding, but this approach is so computationally intensive that it has thus far remained relatively unexplored. Here, we perform MD simulations of sufficient length to observe several different fragments spontaneously and repeatedly bind to and unbind from the protein FKBP, allowing the binding affinities, on- and off-rates, and relative occupancies of alternative binding sites and alternative poses within each binding site to be estimated, thereby illustrating the potential of long time scale MD as a quantitative tool for fragment-based drug discovery. The data from the long time scale fragment binding simulations reported here also provide a useful benchmark for testing alternative computational methods aimed at characterizing fragment binding properties. As an example, we calculated binding affinities for the same fragments using a standard free energy perturbation approach and found that the values agreed with those obtained from the fragment binding simulations within statistical error.


Clinical decision velocity is increased when meta-search filters enhance an evidence retrieval system.
To test whether the use of an evidence retrieval system that uses clinically targeted meta-search filters can enhance the rate at which clinicians make correct decisions, reduce the effort involved in locating evidence, and provide an intuitive match between clinical tasks and search filters. A laboratory experiment under controlled conditions asked 75 clinicians to answer eight randomly sequenced clinical questions, using one of two randomly assigned search engines. The first search engine Quick Clinical (QC) was equipped with meta-search filters (the combined use of meta-search and search filters) designed to answer typical clinical questions e.g., treatment, diagnosis, and the second 'library model' system (LM) offered free access to an identical evidence set with no filter support. Changes in clinical decision making were measured by the proportion of correct post-search answers provided to questions, the time taken to answer questions, and the number of searches and links to documents followed in a search session. The intuitive match between meta-search filters and clinical tasks was measured by the proportion and distribution of filters selected for individual clinical questions. Clinicians in the two groups performed equally well pre-search. Post search answers improved overall by 21%, with 52.2% of answers correct with QC and 54.7% with LM (chi(2) = 0.33, df = 1, p > 0.05). Users of QC obtained a significantly greater percentage of their correct answers within the first two minutes of searching compared to LM users (QC 58.2%; LM 32.9%; chi(2) = 19.203, df = 1, p < 0.001). There was a statistical difference for QC and LM survival curves, which plotted overall time to answer questions, irrespective of answer (Wilcoxon, p = 0.019) and for the average time to provide a correct answer (Wilcoxon, p = 0.006). The QC system users conducted significantly fewer searches per scenario (m = 3.0 SD = 1.15 versus m = 5.5 SD1.97, t = 6.63, df = 72, p = 0.0001). Clinicians using the QC system followed fewer document links than did those who used LM (respectively 3.9 links SD = 1.20 versus 4.7 links SD = 1.79, t = 2.13, df = 72, p = 0.0368). In 6 of the 8 questions, two meta-search filters accounted for 89% or more of clinicians' first choice, suggesting the choice of filter intuitively matched the clinical decision task at hand. Meta-search filters result in clinicians arriving at answers more quickly than unconstrained searches across information sources, and appear to increase the rate with which correct decisions are made. In time restricted clinical settings meta-search filters may thus improve overall decision accuracy, as fewer searches that could otherwise lead to a correct answer are abandoned. Meta-search filters appear to be intuitive to use, suggesting that the simplicity of the user model would fit very well into clinical settings.


Bioimpedance analysis vs. DEXA as a screening tool for osteosarcopenia in lean, overweight and obese Caucasian postmenopausal females.
We aimed at evaluating the efficiency of a newly developed, advanced Bioimpedance Analysis (BIA-ACC®) device as a screening tool for determining the degree of obesity and osteosarcopenia in postmenopausal women with normal or decreased bone density determined by Dual-Energy X-Ray absorptiometry (DEXA) in a representative sample of Greek postmenopausal women. This is a single-gate cross-sectional study of body composition measured by BIA-ACC® and DEXA. Postmenopausal females with BMI ranging from 18.5 to 40 kg/m2 were subjected to two consecutive measurements of DEXA and BIA-ACC® within 5-10 minutes of each other. We used Pearson's co-efficient to examine linear correlations, the intraclass correlation co-efficient (ICC) to test reliability, Bland-Atman plots to assess bias and Deming regressions to establish the agreement in parameters measured by BIA-ACC® and DEXA. Last, we used ANOVA, with Bonferroni correction and Dunnett T3 post hoc tests, for assessing the differences between quantitative and Pearson's x2 between qualitative variables. Our sample consisted of 84 overweight/obese postmenopausal women, aged 39-83 years, of whom 22 had normal bone density, 38 had osteopenia and 24 had osteoporosis based on DEXA measurements, using quota sampling. ICCs and Deming regressions showed strong agreement between BIA-ACC® and DEXA and demonstrated minimal proportional differences of no apparent clinical significance. Bland-Altman plots indicated minimal biases. Fat, skeletal and bone mass measured by BIA-ACC® and DEXA were increased in the non-osteopenic/non-osteoporotic women compared with those of the osteopenic and osteoporotic groups. BIA-ACC® is a rapid, bloodless and useful screening tool for determining body composition adiposity and presence of osteo-sarcopenic features in postmenopausal women. Women with osteopenia and osteoporosis evaluated by DEXA had decreased fat, skeletal and bone mass compared with normal bone density women, suggesting concordance in the change of these three organ masses in postmenopausal women.


[Anterior semicircular canal Benign Paroxysmal Positional Vertigo].
Anterior semicircular canal Benign Paroxysmal Positional Vertigo (antBPPV) is the rarest form of semicircular canalolithiasis, corresponding to about 1-2 % of the BPPV. The diagnosis is obtained by either the Dix-Hallpike maneuver or the straight head hanging positioning maneuver, which provoke a characteristic positional down-beating nystagmus. This vertical nystagmus can be associated with a torsional component that helps in localizing the affected side. The differential diagnosis of antBPPV includes the various central lesions that produce vertical down beating nystagmus (posterior fossa). Several liberatory maneuvers have been proposed for the treatment of antBPPV, but still need standardization.


[Venous malformations and coagulopathy].
Activation of the coagulation process is frequently encountered in patients with venous vascular malformations and results in the local formation of clots and the consumption of components of the coagulation process (platelets, clotting factors). This activation accounts for multiple biological abnormalities such as the elevation of the D-dimers, reduction of fibrinogen and platelets count and less frequently a local intravascular coagulation (LIC). This process seems to be responsible for the local painful symptoms. In case of LIC, bleeding complications may be observed. Therapeutically, antithrombotic treatment with low-molecular weight heparin can improve the local coagulopathy with beneficial effects on blood tests disturbances, pain and bleeding tendency.


The low-temperature study of D- and DL-camphoric anhydride.
The low-temperature crystal stuctures of D- and DL-camphoric anhydride, C(10)H(14)O(3), have been determined by X-ray diffraction methods. Although the two enantiomers crystallize in different space groups, the cell volumes and densities are essentially the same. The six-membered rings deviate significantly from planarity, both exhibiting half-boat conformations. The dihedral angle between the six- and five-membered rings is 80.3 (1) degrees in both cases. The main difference in the molecular stuctures can be described by two torsion angles associated with the H atoms of the methyl substituents. The packing of the racemic and chiral structures are essentially the same.


An Innovative Resource to Guide and Track Medical Resident Activity and Wellness During At-Home Assignments.
Background and objectives Medical residents may work from home for various reasons, including study electives, isolation due to exposure to illness, or during parental leave when they choose to work at home instead of taking extended time off. The University of Arkansas for Medical Sciences (UAMS) Little Rock Family Medicine residency program recognized the need for a tool that provided residents with a list of resources and approved activities for at-home work and a means of tracking their performance in those activities. Methods The administration team at the UAMS Little Rock Family Medicine residency program custom-designed a Daily Activities Log that served multiple purposes. Family medicine residents used it to choose what activities to participate in from a comprehensive list of activities and resources including virtual conferences, recorded didactics, modules, and other online materials. The program provided the framework on the log, while residents used it to document time spent on those activities. The log also gave the program a daily update on the resident's health, as one question specifically asked about well-being. Since it was built in an electronic survey format already owned by the residency program, it was completely customizable and available at no additional cost. Results In the two years covered by this project, residents logged a total of 593 hours of at-home work. In response to a survey, 76% of participating residents (N=14) rated the log as extremely or very easy to use; 64% indicated that it was a helpful resource; and 50% said that it simplified the reporting of their daily status. The residency program faculty found that having one source to track all off-site residents was an efficient means of monitoring their well-being and their work. Conclusions The Daily Activities Log is a versatile tool that provides comprehensive information, resources, and approved activities for residents, documents their work, and provides updates to residency management. It can be readily modified for use in other programs that have residents working at home.


Visceral adipose tissue in patients with severe mental illness.
Background Severe mental illnesses (SMIs), i.e. major depression, schizophrenia and bipolar disorder, are associated with an elevated risk for the development of type-2 diabetes mellitus and cardiovascular disorders. Several factors have been associated with increased cardiometabolic morbidity and mortality in SMI, including lifestyle factors (smoking, inactivity, unhealthy diet), endocrine and immunologic alterations; however, the underlying mechanisms remain to be fully uncovered. It is now well accepted that visceral adipose tissue (VAT) promotes the development of cardiometabolic disorders, at least in part by inflammatory and metabolic functions. Methods This paper reviews studies concerning VAT, with special focus on intra-abdominal and pericardial adipose tissue, in SMI. Results In patients with SMI, several studies have been performed concerning VAT. Most of these studies reported alterations of VAT particularly in patients with major depression and schizophrenia, independent of body weight and body mass index. Some of the studies also reported an increased cardiometabolic risk. Conclusion Patients with SMI are at increased risk of developing cardiometabolic disorders, and display increased amounts of VAT. As studies so far were mainly performed on patients before the onset of cardiometabolic disorders, VAT may serve as a biomarker for patients with SMI to assess cardiometabolic risks beyond established risk scores. Further, interventions aiming at reducing VAT in SMI are highly recommended in long-term multimodal treatment plans.


The expanding role of scapular free-flaps.
The review evaluates the expanding role of scapular free flaps and its application in various complex head and neck defects. The recent literature published focuses on the use of three-dimensional modeling for maxillectomy defects, expanded uses in both the megaflap, the various application of the scapular tip flap, and the feasibility of osseointegrated implants. The subscapular system of flaps remains a versatile option in head and neck reconstruction. It continues to be one of the best options for large defects requiring generous soft tissue, muscle, and bone. Three-dimensional modeling is currently being applied to complex maxillary defects resulting in increased accuracy and efficiency of reconstruction. Expanded uses have also adopted the latissimus dorsi muscle in conjunction with the scapular bony flap to be used in large defects with resulting decreased fistula rates. Finally, there is sufficient evidence that supports the use of dental implantation into the scapula bone with successful retention rates.


Sudomotor and vasomotor activity during the menstrual cycle with global heating.
Many studies have reported that there are changes in sympathetic activity throughout the menstrual cycle as there are oestrogen receptor in the hypothalamus and all other parts of the sympathetic nervous system. The purpose of this study was to see whether there were variations in sympathetic activity, skin vasomotor and sweat gland sudomotor rhythms during the menstrual cycle. Eight young female subjects with a regular menstrual cycle participated in the study. Subjects were tested once during the follicular phase and once during the luteal phase. Skin blood flow and sweat rate were significantly higher in the luteal phase compared with the follicular phase (p < .05), but the frequency and magnitude of sudomotor and vasomotor rhythms were significantly greater in the follicular phase (p < .05). In contrast, spectral data showed less sympathetic activity in the luteal phase. A significant finding here is that the sudomotor rhythm of sweat glands is altered by the menstrual cycle.


The association between polymorphisms in Wnt antagonist genes and bone response to hormone therapy in postmenopausal Korean women.
The aim of this study was to explore the association between polymorphisms in Wnt antagonist genes and bone response to hormone therapy (HT) in postmenopausal Korean women. A prospective study was conducted with 303 postmenopausal women receiving sequential estrogen plus progestogen therapy in a university hospital. The dickkopf (Dkk) 1 c.318A>G, Dkk2 c.437G>A, Dkk3 c.1003A>G, secreted frizzled-related protein (sFRP) 1 rs3242C>T, rs16890444C>T, sFRP3 c.970C>G, sFRP4 c.958C>A, c.1019G>A, and sFRP5 c.20G>C polymorphisms were analyzed, and bone mineral density (BMD) at the lumbar spine and femoral neck (FN) was measured before and after 1 year of sequential estrogen plus progestogen therapy. The percentage changes in BMD of the FN after 1 year of HT were found to be significantly (P < 0.05) different according to the haplotype genotype composed of the sFRP4 c.958C>A and c.1019G>A polymorphisms after adjustment for baseline BMD. The percentage change in BMD at the FN after 1 year of HT was significantly higher in the AA/AG haplotype genotype than in the AG/CG (P < 0.01) or CG/CG (P < 0.05) haplotype genotype. However, any single and combined polymorphisms measured were not related with nonresponsiveness to HT when a nonresponder was defined as a woman who had lost more than 3% of BMD per year after HT. The haplotype genotypes of sFRP4 c.958C>A and c.1019G>A polymorphisms are genetic factors that affect changes in BMD of the FN after HT in postmenopausal Korean women.


Fabrication and Characterization of Novel Food-Grade Bigels Based on Interfacial and Bulk Stabilization.
Novel food-grade bigels were fabricated using zein nanoparticles for interfacial stabilization and non-surfactant gelators (beeswax and tapioca) for bulk stabilization. The present study demonstrated the importance of interfacial stability for biphasic gels and sheds light on the roles of the gelation mechanism and the oil/water ratio of a bigel on its microstructure, physical properties, and digestion behaviors. The results indicated that it is not an easy task to realize homogenization and subsequent gelation in beeswax-tapioca biphasic systems, as no amphiphilic components existed. However, applying the binding of zein nanoparticles at the oil-water interface allowed us to produce a homogeneous and stable bigel (oil fraction reach 40%), which exhibited enhanced structural and functional properties. Oleogel structures play a crucial role in determining the deformation response of bigel systems. As the oil content increased, the mechanical strength and elastic properties of bigels were enhanced. In the meantime, clear bigel-type transitions were observed. In addition, the fabricated bigels were shown to be beneficial for delayed digestion, and the lowest degree of lipolysis could be found in bigel with 50% oleogel.


Long non-coding RNA profiling of pediatric Medulloblastoma.
Medulloblastoma (MB) is one of the most common malignant cancers in children. MB is primarily classified into four subgroups based on molecular and clinical characteristics as (1) WNT (2) Sonic-hedgehog (SHH) (3) Group 3 (4) Group 4. Molecular characteristics used for MB classification are based on genomic and mRNAs profiles. MB subgroups share genomic and mRNA profiles and require multiple molecular markers for differentiation from each other. Long non-coding RNAs (lncRNAs) are more than 200 nucleotide long RNAs and primarily involve in gene regulation at epigenetic and post-transcriptional levels. LncRNAs have been recognized as diagnostic and prognostic markers in several cancers. However, the lncRNA expression profile of MB is unknown. We used the publicly available gene expression datasets for the profiling of lncRNA expression across MB subgroups. Functional analysis of differentially expressed lncRNAs was accomplished by Ingenuity pathway analysis (IPA). In the current study, we have identified and validated the lncRNA expression profile across pediatric MB subgroups and associated molecular pathways. We have also identified the prognostic significance of lncRNAs and unique lncRNAs associated with each MB subgroup. Identified lncRNAs can be used as single biomarkers for molecular identification of MB subgroups that warrant further investigation and functional validation.


Improving best-phase image quality in cardiac CT by motion correction with MAM optimization.
Research in image reconstruction for cardiac CT aims at using motion correction algorithms to improve the image quality of the coronary arteries. The key to those algorithms is motion estimation, which is currently based on 3-D/3-D registration to align the structures of interest in images acquired in multiple heart phases. The need for an extended scan data range covering several heart phases is critical in terms of radiation dose to the patient and limits the clinical potential of the method. Furthermore, literature reports only slight quality improvements of the motion corrected images when compared to the most quiet phase (best-phase) that was actually used for motion estimation. In this paper a motion estimation algorithm is proposed which does not require an extended scan range but works with a short scan data interval, and which markedly improves the best-phase image quality. Motion estimation is based on the definition of motion artifact metrics (MAM) to quantify motion artifacts in a 3-D reconstructed image volume. The authors use two different MAMs, entropy, and positivity. By adjusting the motion field parameters, the MAM of the resulting motion-compensated reconstruction is optimized using a gradient descent procedure. In this way motion artifacts are minimized. For a fast and practical implementation, only analytical methods are used for motion estimation and compensation. Both the MAM-optimization and a 3-D/3-D registration-based motion estimation algorithm were investigated by means of a computer-simulated vessel with a cardiac motion profile. Image quality was evaluated using normalized cross-correlation (NCC) with the ground truth template and root-mean-square deviation (RMSD). Four coronary CT angiography patient cases were reconstructed to evaluate the clinical performance of the proposed method. For the MAM-approach, the best-phase image quality could be improved for all investigated heart phases, with a maximum improvement of the NCC value by 100% and of the RMSD value by 81%. The corresponding maximum improvements for the registration-based approach were 20% and 40%. In phases with very rapid motion the registration-based algorithm obtained better image quality, while the image quality of the MAM algorithm was superior in phases with less motion. The image quality improvement of the MAM optimization was visually confirmed for the different clinical cases. The proposed method allows a software-based best-phase image quality improvement in coronary CT angiography. A short scan data interval at the target heart phase is sufficient, no additional scan data in other cardiac phases are required. The algorithm is therefore directly applicable to any standard cardiac CT acquisition protocol.


Simulations of frequency-dependent photoacoustic magnitude signals and their implications for bacteriorhodopsin photocycle energetics.
The modulation frequency dependence of photoacoustic signals obtained from photoactive samples can provide information on the time-dependent enthalpy changes occurring during the light-induced process. The experimental requirements for this type of calorimetry, and the interpretation ot the results, are critically examined with reference to the light-driven proton pump bacteriorhodopsin. For a three-step unbranched model of the bacteriorhodopsin photocycle we derive an expression for the photoacoustic magnitude signal as a function of frequency. Simulations are performed for various values of the rate constants and energetic changes. It is shown that the net heat uptake during a low, final step postulated by some workers should be reflected in the photoacoustic magnitude frequency spectrum, giving rise to a characteristic maximum. However, this effect, which has been observed experimentally, may also be produced by a fast, strongly endothermic step occurring earlier. The precise chronology of an endothermic transition cannot be resolved unambiguously by magnitude measurements alone, although they are free from assumptions regarding difficult-to-measure phase relationships. Hence, the published photoacoustic observations showing the effect are consistent with a cyclic sequence of events in which the bacteriorhodopsin system first undergoes an increase of entropy, followed by a decrease on returning to the initial state, as well as the reverse. It is argued that the molecular disorder-order sequence is more probable.


Solvation dynamics of a single water molecule probed by infrared spectra--theory meets experiment.
The dynamics and energetics of water at interfaces or in biological systems plays a fundamental role in all solvation and biological phenomena in aqueous solution. In particular, the migration of water molecules is the first step that controls the overall process in the time domain. Experimentally, the dynamics of individual water molecules is nearly impossible to follow in solution, because signals from molecules in heterogeneous environments overlap. Although molecular dynamics simulations do not have this restriction, there is a lack of experimental data to validate the calculated dynamics. Here, we demonstrate a new strategy, in which the calculated dynamics are verified by measured time-resolved infrared spectra. The coexistence of fast and slow migrations of water molecules around a CONH peptide linkage is revealed for a model system representative of a hydrate peptide.


The use of the Internet and information technology to facilitate teaching evidence based practice - a case study.
The use of Internet and information technology for teaching nurses and other health professionals has been shown to improve learning outcomes, learners' satisfaction and the learner-learner and learner-teacher interactions. Two recent government white pa pers emphasised the central role of evidence based practice as the key component for nurses to deliver quality care to their patients, and basic IT and Internet competencies are also essential for evidence based practice. This paper reports the experience of the introduction of a dedicated web site, the Internet and information technology to augment a face-to-face Master's unit in evidence based practice. The main objective of the unit is to enable students to apply the principles of evidence based practice to their daily professional practice. The organisation of this module and the ways these new technologies were exploited are described. A brief evaluation of this module is presented: the unit was reasonably well received by the students. The practical difficulties of introducing such technologies by individual tutors and ensuring that students have adequate access to computers are discussed.


The effects of detoxification of domestic gas on suicide in the United States.
As domestic gas was detoxified in the United States, the rate of suicide by domestic gas decreased. During this time period (1950-60), there was a parallel increase in the per capita ownership of cars and an accompanying increase in the rate of suicide by motor vehicle exhaust. However, displacement of suicide method from domestic gas to car exhaust occurred only for males and not for females.


Prophylactic antipropagation stitch in total knee arthroplasty performed via medial parapatellar approach: a simple and cost-effective technique for reducing quadriceps injury.
This article describes a novel, simple, and effective technique for limiting extensor mechanism damage during total knee arthroplasty (TKA) performed via the medial parapatellar (MPP) approach. Immediately on making the quadriceps tendon split in the MPP approach, a figure-of-eight stitch is placed at the apex of the split. The prophylactic stitch technique has been used by one surgeon in 1000 consecutive TKAs over the past 5 years. This technique has eliminated proximal extension of the quadriceps tendon split into the muscle belly and may help to decrease postoperative pain, increase postoperative quadriceps strength, and improve patient satisfaction and outcomes.


Ti4O7/g-C3N4 Visible Light Photocatalytic Performance on Hypophosphite Oxidation: Effect of Annealing Temperature.
The oxidation of hypophosphite to phosphate is the key to recover the phosphorus resource from the hypophosphite wastewater. In the present work, Ti4O7/g-C3N4 composites were synthesized at two different temperatures (100 and 160°C) and their performance on photocatalytic oxidation of hypophosphite under visible light irradiation and the corresponding mechanism were evaluated. A hydrolysis method using g-C3N4 and Ti4O7 was applied to synthesize the Ti4O7/g-C3N4 composites with their hybrid structure and morphology confirmed by X-ray diffraction (XRD), scanning electron microscopy (SEM), and X-ray photoelectron spectra (XPS). The annealing temperature significantly affected the photocatalytic performance of Ti4O7/g-C3N4 that the 160-Ti4O7/g-C3N4 composite (fabricated at 160°C) showed the highest oxidation efficiency of hypophosphite of 81% and the highest photocatalytic oxidation rate of 0.467 h-1 comparing with the 100-Ti4O7/g-C3N4 composite (fabricated at 100°C) and pure g-C3N4. The enhanced photocatalytic performance of 160-Ti4O7/g-C3N4 could be ascribed to the effective charge separation and enhanced photoabsorption efficiency. Additionally, electron spin resonance (ESR) results showed that hydroxyl radicals and superoxide anion radicals were mainly responsible to the oxidation of hypophosphite with superoxide anion radicals accounting for a more significant contribution. Moreover, Ti4O7/g-C3N4 photocatalysts showed the remarkable stability in the repetitive experiments.


Standardization of serum neutralization assay of Japanese encephalitis virus (Nakayama NIH strain) on BHK-21 (Cl-13) cell line.
Potency testing of Japanese encephalitis (JE) vaccine has been a complex process since its inception. To overcome difficulties encountered therein, an alternative assay, serum neutralization test (SNT), using Baby Hamster Kidney 21 cell line, has been standardized. The antibody response generated against JE vaccine was quantified and the assay was found to be sensitive and specific enough with significant accuracy and precision. On analysis of cell count, a cell concentration of 1.5 x 104 was selected as the optimum, since concentrations above and below this resulted in problems of confluent monolayer formation and incomplete monolayer formation. Incubation time has also been standardized for measuring cytopathic effect (CPE). Out of the four different time points selected, 90 min was found to be adequate for 50% reduction in the amount of CPE. The accuracy of SNT assay is explained in terms of fiducial limits at 95% level. Inter- and intra-assay reproducibility testing was also performed. A comparison of potency of JE vaccine by plaque reduction neutralization test (PRNT) and SNT method was conducted and it was found that SNT can be a reliable approach for estimating the potency of JE vaccine. The results of this study throw a light on the utility of SNT assay for the potency estimation of JE vaccine in routine practice.


Human Dendritic Cells Synergistically Activated by FVIII Plus LPS Induce Activation of Autologous CD4+ T Cells.
The most severe side effect in haemophilia A (HA) treatment is the development of anti-factor VIII antibodies, also called inhibitors. Why inhibitors develop in a proportion of treated HA patients and how this can be prevented remains largely unanswered. Among numerous theories, the presence of immunological danger signals, associated with events such as surgery or infection, has been proposed to play a role. In this study, we demonstrate that human dendritic cells (DC) synergistically activated by a combination of factor VIII (FVIII) concentrate plus the bacterial danger signal lipopolysaccharide (LPS) induce a significantly stronger activation of autologous CD4+ T cells than DC pretreated with FVIII or LPS alone. The observed T cell activation is dependent on antigen processing, presentation on MHC class II molecules and costimulation via CD86. Of note, FVIII plus LPS pretreated DC predominantly induce the activation of memory T cells and a minor proportion of naive T cells. Collectively, our data support a model in which immunological danger signals plus FVIII concentrates synergistically increase human CD4+ T cell responses to FVIII protein.


Usefulness of C-reactive protein/albumin ratio as a predictor of new-onset atrial fibrillation in SARS-COV-2.
Aim: To investigate whether C-reactive protein/albumin ratio (CAR) has an association with new onset atrial fibrillation (NOAF) in SARS-CoV-2. Materials & methods: This study included 782 patients with SARS-CoV-2 infection, who were hospitalized in Turkey. The end point of the study was an occurrence of NOAF. Results: NOAF was identified in 41 patients (5.2%). Subjects who developed NOAF had a higher CAR compared with those who did not develop NOAF (p < 0.001). In the multivariate logistic regression analysis the CAR (odds ratio = 2.879; 95% CI: 1.063-7.793; p = 0.037) was an independent predictor of NOAF. Conclusion: A high level of CAR in blood samples is associated with an increased risk of developing NOAF in SARS-CoV-2.


Transgenic expression of Fas in T cells blocks lymphoproliferation but not autoimmune disease in MRL-lpr mice.
Fas is a member of the TNF receptor family. Binding of Fas ligand to Fas induces apoptosis in Fas-bearing cells. Fas is expressed in various cells, including thymocytes, peripheral T cells, and activated B cells. The mouse lpr mutation is a loss of function mutation of Fas. MRL-lpr/lpr mice develop lymphadenopathy and splenomegaly, and produce multiple autoantibodies, which results in autoimmune disease. In this report, we describe the establishment of a line of Fas transgenic MRL-lpr mice in which mouse Fas cDNA was expressed using the T cell-specific murine lck promoter. The transgenic mice expressed functional Fas in thymocytes and peripheral T cells, but not in B cells. The transgenic mice did not accumulate abnormal T cells (Thy-1+ B220+), but still accumulated B cells (Thy-1- B220+); they produced a large quantity of Igs (IgG1 and IgG2a), including anti-DNA Abs, and developed glomerulonephritis. These results suggest that autoreactive or activated B cells must be killed through Fas expressed in the B cells by the Fas ligand expressed in activated T cells.


Inactivation of parkin by promoter methylation correlated with lymph node metastasis and genomic instability in nasopharyngeal carcinoma.
This study aimed to investigate the inactivation of the parkin gene by promoter methylation and its relationship with genome instability in nasopharyngeal carcinoma. Parkin was considered as a tumor suppressor gene in various types of cancers. However, its role in nasopharyngeal carcinoma is unexplored. Genomic instabilities were detected in nasopharyngeal carcinoma tissues by the random amplified polymorphic DNA. The methylation-specific polymerase chain reaction, semi-quantitative reverse transcription polymerase chain reaction, and immunohistochemical analysis were used to detect methylation and mRNA and protein expression of parkin in 54 cases of nasopharyngeal carcinoma tissues and 16 cases of normal nasopharyngeal epithelia tissues, and in 5 nasopharyngeal carcinoma cell lines (CNE1, CNE2, TWO3, C666, and HONE1) and 1 normal nasopharyngeal epithelia cell line (NP69). mRNA expression of parkin in CNE1 and CNE2 was analyzed before and after methyltransferase inhibitor 5-aza-2-deoxycytidine treatment. The relationship between promoter methylation and mRNA expression, demethylation and mRNA expression, and mRNA and protein expression of the gene and clinical factors and genomic instabilities were analyzed. The mRNA and protein expression levels were significantly reduced in 54 cases of human nasopharyngeal carcinoma compared with 16 cases of normal nasopharyngeal epithelia. Parkin-methylated cases showed significantly lower mRNA and protein expression levels compared with unmethylated cases. After 5-aza-2-deoxycytidine treatment, parkin mRNA expression was restored in CNE1 and CNE2; 92.59% (50/54) of nasopharyngeal carcinoma demonstrated genomic instability. Parkin is frequently inactivated by promoter methylation, and its mRNA and protein expression correlate with lymph node metastasis and genomic instability. Parkin deficiency probably promotes tumorigenesis in nasopharyngeal carcinoma.


The Protective Effect of Chrysanthemum indicum Extract against Ankylosing Spondylitis in Mouse Models.
In traditional Chinese and Korean homeopathic medicine, Chrysanthemum indicum Linné (Asteraceae) is a time-honored herb, prescribed for the resolution of symptoms associated with inflammatory and hypertensive conditions as well as those affecting the lungs and its associated structures. The goal of this work is to investigate the defensive role of Chrysanthemum indicum extract in fighting ankylosing spondylitis (AS) using mouse models, through which the manifestation and extent of the disease progression were measured with quantitative analysis of the intervertebral joints. Markers of inflammation as well as oxidative stress were also analysed. Western blot was used to quantify the levels of Nuclear Factor-κB (NF-κB) p65, Dickkopf-1 (DKK-1), and sclerostin (SOST). Consequently, the findings of this experiment demonstrated that AS in mice that were given Chrysanthemum indicum extract had lower level of TNF-α, IL-1β, and IL-6 (P < 0.05) and increased level of catalase (CAT), glutathione peroxidase (GSH-PX), and superoxide dismutase (SOD) (P < 0.05). The results also revealed that Chrysanthemum indicum supplemented with diet contributed to a decrease in Nuclear Factor-κB (NF-κB) p65 protein expression (P < 0.05) and higher levels of DKK-1 and SOST proteins (P < 0.05). Therefore, we concluded that the beneficial role of Chrysanthemum indicum in AS is manifested through downregulating oxidative stress, inhibiting inflammatory mediators and NF-κB, and increasing DKK-1 and SOST levels.


Implications of exercise testing for prediction of athletic performance: a contemporary perspective.
One of the most fundamental beliefs in exercise physiology is that performance during maximum exercise of short duration is limited by the inability of the heart and lungs to provide oxygen at a rate sufficiently fast to fuel energy production by the active muscle mass. This belief originates from work undertaken in the 1920's by Hill and Lupton. A result is that most, if not all, of the studies explaining the effects of exercise training or detraining or other interventions on human physiology explain these changes in terms either of central adaptations increasing oxygen delivery to muscle or of peripheral adaptations that modify the rates of oxygen or fuel utilization by the active muscles. Yet a critical review of Hill and Lupton's results shows that they inferred but certainly did not prove that oxygen limitation develops during maximal exercise. Furthermore, more modern studies suggest that, if such an oxygen limitation does indeed occur during maximal exercise, it develops in about 50% of test subjects. Thus, an alternative mechanism may need to be evoked to explain exhaustion during maximal exercise in a rather large group of subjects. This review proposes that the factors limiting maximal exercise performance might be better explained in terms of a failure of muscle contractility ("muscle power"), which may be independent of tissue oxygen deficiency. The implications for exercise testing and the prediction of athletic performance are discussed.


[Significance of type III serum N-terminal peptide procollagen assay in chronic viral hepatitis C].
The aim of this study was to evaluate the relationship between the serum N-terminal peptide of type III procollagen (PIIINP), Knodell's score and the response to treatment in 79 patients with viral C chronic hepatitis. Liver biopsy and serum PIIINP was assessed in all patients. Serum PIIINP was correlated with Knodell'score (r = 0.60; P = 10(-4)). Correlation between serum alanine amino-transferase and Knodell'score was smaller than for PIIINP (r = 0.38; P = 0.02). Forty-nine patients were treated with interferon alpha. A second biopsy and serum PIIINP determination were performed one year after treatment discontinuation. Means of serum PIIINP and Knodell'score significantly decreased after treatment (P < 0.01; P < 0.03). Among non-responder patients, serum PIIINP was initially more elevated: 37.5 +/- 12.6 ng/mL versus 22.6 +/- 3.8 (P < 10(-3)). Knodell's score decreased only among responder patients (P < 0.01 vs P < 0.7). Positive and negative predictive value for the response to treatment, evaluated by aminotransferases normalization, of serum PIIINP level < or = 24 ng/mL were 0.6 and 0.9 respectively. These findings suggest that serum PIIINP is a relevant marker of activity in patients with viral C chronic hepatitis and that it might have a predictive value of response to treatment.


Fibropapillomatosis in a Green Sea Turtle ( Chelonia mydas) from the Southeastern Pacific.
Fibropapillomatosis is a neoplastic disease that afflicts sea turtles. Although it is disseminated worldwide, cases of the disease have not been reported in the southeastern Pacific region. We describe a case of fibropapillomatosis in a green sea turtle ( Chelonia mydas) during its rehabilitation at the Machalilla National Park Rehabilitation Center, Ecuador. Viral presence was confirmed by PCR, targeting fragments of the chelonid alphaherpesvirus 5 (ChHV5) unique long (UL) genes, UL27, UL28, and UL30. The amplicons were sequenced and included in a global phylogenetic analysis of the virus with other reported sequences from GenBank. Results showed that the available viral sequences segregated into five phylogeographic groups: western Atlantic and eastern Caribbean, central Pacific, western Pacific, Atlantic, and eastern Pacific groups. The concatenated ChHV5 sequences from Ecuador clustered with the eastern Pacific sequences.


An Exploratory Study on the Impact of Domestic Violence on the Families in Trinidad and Tobago, the West Indies.
The purpose of this study is to explore the impact of domestic violence on the economic condition of the families. This cross-sectional study utilized a non-probability sampling procedure (purposive sampling) that included 30 women who have sought services from the Coalition Against Domestic Violence Agency. Data were collected using a questionnaire, which was comprised of 21 questions. The questions sought information on socioeconomic conditions and impact on domestic violence on the financial position. The study revealed that more of domestic violence victims were at an early age. Recommendations for future research include identifying the major causes for family disorganization and break down in the families arise out of domestic violence and other associated factors where explored while emphasizing the importance of family-based programs that minimize the impact.


Nestling telomere length does not predict longevity, but covaries with adult body size in wild barn swallows.
Telomere length and dynamics are increasingly scrutinized as ultimate determinants of performance, including age-dependent mortality and fecundity. Few studies have investigated longevity in relation to telomere length (TL) in the wild and none has analysed longevity in relation to TL soon after hatching, despite the fact that telomere shortening may mostly occur early in life. We show that TL in nestling barn swallows (Hirundo rustica) in the wild does not predict longevity. However, TL positively covaries with body size, suggesting that individuals with large TL can afford to grow larger without paying the cost of reduced TL, and/or that benign rearing conditions ensure both large body size and low rates of telomere shortening. Overall, our study hints at a role of TL in developmental processes, but also indicates a need for further analyses to assess the expectation that TL in young individuals predicts longevity in the wild.


Spine Stereotactic Body Radiotherapy for Prostate Cancer Metastases and the Impact of Hormone Sensitivity Status on Local Control.
Stereotactic body radiotherapy (SBRT) is used to deliver ablative dose of radiation to spinal metastases. To report the first dedicated series of spine SBRT specific to prostate cancer (PCa) metastases with outcomes reported according to hormone sensitivity status. A prospective database was reviewed identifying patients with PCa treated with spine SBRT. This included those with hormone-sensitive PCa (HSPC) and castrate-resistant PCa (CRPC). The primary end point was MRI-based local control (LC). A total of 183 spine segments in 93 patients were identified; 146 segments had no prior radiation and 37 had been previously radiated; 27 segments were postoperative. The median follow-up was 31 months. At the time of SBRT, 50 patients had HSPC and the remaining 43 had CRPC. The most common fractionation scheme was 24-28 Gy in 2 SBRT fractions (76%). LC rates at 1 and 2 years were 99% and 95% and 94% and 78% for the HSPC and CRPC cohorts, respectively. For patients treated with de novo SBRT, a higher risk of local failure was observed in patients with CRPC (P = .0425). The 1-year and 2-year overall survival rates were significantly longer at 98% and 95% in the HSPC cohort compared with 79% and 65% in the CRPC cohort (P = .0005). The cumulative risk of vertebral compression fracture at 2 years was 10%. Favorable LC rates were observed after spine SBRT for PCa metastases; strategies to improve long-term LC in patients with CRPC require further investigation.


Simple, reversible gradient Seebeck coefficient measurement system for 300-600 K with COMSOL simulations.
Seebeck measurement is a crucial step for characterizing thermoelectric samples, as measuring the accurate value with a simpler system design is challenging. Here, we report a simple design of the Seebeck coefficient measurement system, which can measure the thermo-emf (Seebeck coefficient) of the sample, under a limited temperature range of 300-600 K. Unlike the majority of the reported instrumental designs, the system does not have a hot walled chamber. The sample is sandwiched between two brass block supported heaters, which are controlled separately. Thus, this type of system is suitable for a window of the temperature range near room temperature. In this paper, we report the system that can measure the Seebeck coefficient up to 600 K. The heaters touch the sample through 1 mm thick silver caps, which offer insignificant thermal resistance and a stable temperature, as seen through experiment as well as COMSOL simulations. A typical sample has, at maximum, a diameter of 10 mm and a thickness of 2-3 mm. A reversible temperature gradient is applied in quasi-static direct current mode. By virtue of its design, the sample holder ensures a minimum thermal and electrical contact resistance during a measurement cycle. The combination of metals used for measurement (Ag and Cu) shows negligible junction contribution. The variance up to ±2% and accuracy up to 8% at a high temperature have been obtained using calibration sample reference data of state-of-the-art commercial systems.


[Strengthening the interdisciplinary process in student's research programs].
The modern disciplinary focus is based on specific processes and presents a tendency to reduce knowledge by describing it as a consequence of a highly rational scientific posture that has promoted excessive specialization and fragmentation in detriment of the human being. With the development of societies and the perspective of knowledge interconnection which searches for institutions with global and broad dimensions in the spheres of knowing and doing, disciplinarity will be transcended. In this sense, the purpose of this work is to report the experience of an undergraduate students' research program in the view of interdisciplinarity. Playing the roles of advisor and student from different areas--Nursing and Administration respectively--we are experiencing an interdisciplinary process through the exchange of ideas, actions and the investigation on human resources and their management in the Nursing area. We believe that the dissemination of this experience can contribute to strengthen interdisciplinarity in the context of research in undergraduate programs.


Mitochondrial gene expression in Saccharomyces cerevisiae. IV. Effects of yeast cytosol on mitochondrial protein synthesis, degradation, and respiration.
It has been known for some time that the addition of a crude yeast cytosolic fraction to isolated mitochondria stimulates the rate of amino acid incorporation into protein in the isolated organelles. However, the mechanism and importance of this phenomenon relative to mitochondrial function has not been established. While it has been assumed that this effect is at the level of translation, the recognition that newly synthesized mitochondrial translation products are rapidly degraded in isolated yeast mitochondria raises the possibility that cytosol affects amino acid incorporation by inhibiting proteolysis. Using pulse-chase experiments we demonstrate that the rate constants of degradation of the nascent products are not affected by yeast cytosol. Further, not only is proteolysis not inhibited by cytosol, but the loss of label caused by proteolysis is actually increased. This increase is directly related to an increase in the size of the nascent product pool which increases simply as a consequence of increasing the rate of translation. By utilizing an approach in which the loss of label due to proteolysis is minimized, the true stimulatory activity of the cytosolic fraction on synthesis was determined (2.1-fold vs. 1.3-fold by the previous method). Pulse-chase experiments in the presence of pactamycin, an initiation inhibitor, demonstrate that yeast cytosol causes an initial increase in the rate of translational initiation without increasing the rate of elongation. However, at later intervals the yeast cytosol acts primarily to maintain the rate of elongation which falls steadily in the controls. Finally, the presence of yeast cytosol dramatically increases the length of incubation time in which the mitochondrial preparation consumes oxygen and maintains coupled respiration, parameters that fall rapidly in the controls. Thus, a yeast cytosolic fraction may function to promote the stability of the mitochondrial preparation, which in turn may account for the increase in rates of translation, particularly with regard to maintaining rates of elongation.


A physiologically based pharmacokinetic modelling approach to predict buprenorphine pharmacokinetics following intravenous and sublingual administration.
Opioid dependence is associated with high morbidity and mortality. Buprenorphine (BUP) is approved by the Food and Drug Administration to treat opioid dependence. There is a lack of clear consensus on the appropriate dosing of BUP due to interpatient physiological differences in absorption/disposition, subjective response assessment and other patient comorbidities. The objective of the present study was to build and validate robust physiologically based pharmacokinetic (PBPK) models for intravenous (IV) and sublingual (SL) BUP as a first step to optimizing BUP pharmacotherapy. BUP-PBPK modelling and simulations were performed using SimCyp® by incorporating the physiochemical properties of BUP, establishing intersystem extrapolation factors-based in vitro-in-vivo extrapolation (IVIVE) methods to extrapolate in vitro enzyme activity data, and using tissue-specific plasma partition coefficient estimations. Published data on IV and SL-BUP in opioid-dependent and non-opioid-dependent patients were used to build the models. Fourteen model-naïve BUP-PK datasets were used for inter- and intrastudy validations. The IV and SL-BUP-PBPK models developed were robust in predicting the multicompartment disposition of BUP over a dosing range of 0.3-32 mg. Predicted plasma concentration-time profiles in virtual patients were consistent with reported data across five single-dose IV, five single-dose SL and four multiple dose SL studies. All PK parameter predictions were within 75-137% of the corresponding observed data. The model developed predicted the brain exposure of BUP to be about four times higher than that of BUP in plasma. The validated PBPK models will be used in future studies to predict BUP plasma and brain concentrations based on the varying demographic, physiological and pathological characteristics of patients.


Motivations for contributing to health-related articles on Wikipedia: an interview study.
Wikipedia is one of the most accessed sources of health information online. The current English-language Wikipedia contains more than 28,000 articles pertaining to health. The aim was to characterize individuals' motivations for contributing to health content on the English-language Wikipedia. A set of health-related articles were randomly selected and recent contributors invited to complete an online questionnaire and follow-up interview (by Skype, by email, or face-to-face). Interviews were transcribed and analyzed using thematic analysis and a realist grounded theory approach. A total of 32 Wikipedians (31 men) completed the questionnaire and 17 were interviewed. Those completing the questionnaire had a mean age of 39 (range 12-59) years; 16 had a postgraduate qualification, 10 had or were currently studying for an undergraduate qualification, 3 had no more than secondary education, and 3 were still in secondary education. In all, 15 were currently working in a health-related field (primarily clinicians). The median period for which they have been an active editing Wikipedia was 3-5 years. Of this group, 12 were in the United States, 6 were in the United Kingdom, 4 were in Canada, and the remainder from another 8 countries. Two-thirds spoke more than 1 language and 90% (29/32) were also active contributors in domains other than health. Wikipedians in this study were identified as health professionals, professionals with specific health interests, students, and individuals with health problems. Based on the interviews, their motivations for editing health-related content were summarized in 5 strongly interrelated categories: education (learning about subjects by editing articles), help (wanting to improve and maintain Wikipedia), responsibility (responsibility, often a professional responsibility, to provide good quality health information to readers), fulfillment (editing Wikipedia as a fun, relaxing, engaging, and rewarding activity), and positive attitude to Wikipedia (belief in the value of Wikipedia). An additional factor, hostility (from other contributors), was identified that negatively affected Wikipedians' motivations. Contributions to Wikipedia's health-related content in this study were made by both health specialists and laypeople of varying editorial skills. Their motivations for contributing stem from an inherent drive based on values, standards, and beliefs. It became apparent that the community who most actively monitor and edit health-related articles is very small. Although some contributors correspond to a model of "knowledge philanthropists," others were focused on maintaining articles (improving spelling and grammar, organization, and handling vandalism). There is a need for more people to be involved in Wikipedia's health-related content.


Combined fascia and mesh closure of large incisional hernias.
Large incisional hernias of the abdominal wall represent substantial defects of supportive tissues. The repair of these requires the mobilization of fascia or the use of a prosthetic mesh. A method for closing large midline incisional hernias using both the fascia and a mesh was described in 1979. This repair was used for six midline hernias and four large incisional hernias in the right subcostal region. No wound complications and no recurrences (median follow-up 1 year 5 months) were seen. The combined fascia and mesh repair can be successfully used for large incisional hernias of the anterior abdominal wall in areas other than the midline.


Stereotactic exploration of brain tumours by ultrasound.
A report upon a new technique for investigation of brain tumour anatomy by means of stereotactically directed ultrasound is presented with some illustrative cases. Information obtained with this technique has been found useful when deciding the treatment program for the tumour.


Use of a high frequency transducer with real time B-mode ultrasound scanning to identify early pregnancy in cows.
The reproductive tracts of 22 Friesian dairy cows were examined from seven to 35 days after insemination using a real time B-mode ultrasound scanner with a 7.5 MHz transducer. The earliest detection of pregnancy was at nine days when a vesicle was imaged within the lumen of the uterine horn. The early conceptus was seen at day 13 within the vesicle and these structures were followed ultrasonically until day 35. There was a sudden enlargement of the vesicle at day 19 and a heart beat was detected in the embryo at day 22. The allantois was imaged at day 23 and the amnion by day 29. The embryonic outline was clearly defined by day 33 when the body cavities could be discerned. This ability to determine pregnancy at an early stage should prove to be a useful technique in investigating the problems associated with early embryonic death in cattle.


[The influence of the urethral pressure profile on the prognosis of incontinence surgery (author's transl)].
In 114 out of 1215 stress-incontinent women the maximal urethral closure pressure was determined. Descensus of urinary bladder and urethra was proved by lateral urethrocystography. In the group of sphincter weakness (Edwards) which was to be found in 46 patients, only 48% became continent after colporrhaphy, Marshall-Marchetti-Krantz or sling surgery. If the pressure profile was found to be normal 74% of our patients felt cured after incontinence-operation. Sphincter weakness determines a failure due to surgery which is statistically of significance (P less than 0,01).


Nothogreniera new genus, for two species of Australian "Paracnephia" (Diptera: Simuliidae).
Two species of Australian Simuliidae known only from adult females and currently assigned to "Paracnephia" are re-described, as are their now-known males and immature stages. Morphological character states of "Paracnephia" fergusoni (Tonnoir) and "P." fergusoni var. (Mackerras Mackerras) reveal that they are markedly distinct from all other Australian species, and are here assigned to the new genus-Nothogreniera-the most plesiomorphic Gondwanan Australian simuliid. Structural variation among populations of N. fergusoni suggests that this entity comprises a species complex.


Detection of a metalloproteinase in patients with acute and chronic renal failure.
The effect of different dialyzer membrane materials (cuprophan, cellulose hydrate, polyacrylonitrile, polymethylmethacrylate, ethylene-vinyl alcohol copolymer) on the ultrafiltrate proteinase activity was investigated in 26 patients with acute renal failure (ARF) and 40 patients undergoing regular hemodialysis treatment (RDT). Furthermore, the proteinase activity was characterized in vitro using azocasein and phosphorylase kinase as substrates in the absence and presence of different proteinase inhibitors. Proteinase activity of ultrafiltrates obtained from ARF patients was significantly enhanced with the dialyzer KF 101 (ethylene-vinyl alcohol copolymer). The digestion pattern of phosphorylase kinase revealed an identical type of proteinases in ultrafiltrates of ARF and RDT patients. The pH optimum of this proteinase was at alkaline pH. The proteinase activity could be inhibited in the presence of EDTA, whereas serine proteinase inhibitors were ineffective. Furthermore, the inactivated proteinase after Sephadex G-10 chromatography (in order to separate ultrafiltrate electrolytes and trace elements from protein) could be reactivated after the addition of Mg++ and/or Ca++. We conclude that a metalloproteinase can be found in ARF and RDT patients, and that KF 101 is more effectively eliminating the proteinase activity in ARF patients than other dialyzer membranes.


Distinct DNA methylation alterations are associated with cribriform architecture and intraductal carcinoma in Gleason pattern 4 prostate tumors.
The aim of the present study was to explore DNA methylation aberrations in association with cribriform architecture and intraductal carcinoma (IDC) of the prostate, as there is robust evidence that these morphological features are associated with aggressive disease and have significant clinical implications. Herein, the associations of a panel of seven known prognostic DNA methylation biomarkers with cribriform and IDC features were examined in a series of 91 Gleason pattern (GP) 4 tumors derived from Gleason score 7 radical prostatectomies. Gene specific DNA methylation was compared between cribriform and/or IDC positive vs. negative cases, and in association with clinicopathological features, using Chi square and Mann-Whitney U tests. DNA methylation of the adenomatous polyposis coli, Ras association domain family member 1 and T-box 15 genes was significantly elevated in GP4 tumors with cribriform and/or IDC features compared with negative cases (P=0.045, P=0.007 and P=0.013, respectively). To the best of our knowledge, this provides the first evidence for an association between cribriform and/or IDC and methylation biomarkers, and warrants further investigation of additional DNA methylation events in association with various architectural patterns in prostate cancer.


Human and Swine Dental Pulp Stem Cells Form a Vascularlike Network after Angiogenic Differentiation in Comparison with Endothelial Cells: A Quantitative Analysis.
The aim of this study was to quantify vascular network formation capacity after angiogenic induction of human and swine dental pulp stem cells (DPSCs) in comparison with endothelial cells. Primary human DPSCs or swine DPSCs were induced in endothelial growth medium for 7 days. The expression of the endothelial marker von Willebrand factor was determined by immunostaining. Induced DPSCs (iDPSCs) and noninduced DPSCs (niDPSCs) were seeded at different cell numbers onto Matrigel (BD Biosciences, San Jose, CA) for vascular network formation assays and analyzed after 4, 8, 12, and 18 hours in comparison with human microvascular endothelial cells (hMECs). Quantitative analysis of vascular tubule formation was performed using ImageJ software (National Institutes of Health, Bethesda, MD). The vascular network formation was also conducted by coculturing of niDPSCs and iDPSCs. Von Willebrand factor was detected by immunofluorescence in both niDPSCs and iDPSCs (human and swine). Time-lapse microscopic observation showed that the vascular network was formed by iDPSCs but not niDPSCs. After 4 hours, iDPSCs showed vascular network formation, whereas niDPSCs started to aggregate and formed clusters. Human iDPSCs displayed a similar capacity to form vascular networks in Matrigel compared with hMECs based on quantitative analysis. Swine iDPSCs had a higher capacity compared with human iDPSCs or hMECs (P < .05) in forming the network structures including segments, nodes, and mesh. A coculture experiment showed that human niDPSCs colocalized on the angiogenic tubules and vascular networks that were formed by human iDPSCs. Our findings indicate that iDPSCs in combination with their noninduced counterparts may be used as a future clinical strategy for enhancing angiogenesis during the process of pulp-dentin regeneration.


Obesity Management in Kidney Transplant Candidates: Current Paradigms and Gaps in Knowledge.
In this review, we discuss the increasing prevalence of obesity among people with chronic and end-stage kidney disease (ESKD) and implications for kidney transplant (KT) candidate selection and management. Although people with obesity and ESKD receive survival and quality-of-life benefits from KT, most KT programs maintain strict body mass index (BMI) cutoffs to determine transplant eligibility. However, BMI does not distinguish between visceral adiposity, which confers higher cardiovascular risks and risks of perioperative and adverse posttransplant outcomes, and muscle mass, which is protective in ESKD. Furthermore, requirements for patients with obesity to lose weight before KT should be balanced with the findings of numerous studies that show weight loss is a risk factor for death among patients with ESKD, independent of starting BMI. Data suggest that KT is associated with survival benefits relative to remaining on dialysis for candidates with obesity although recipients without obesity have higher delayed graft function rates and longer transplant hospitalization durations. Research is needed to determine the optimal body composition metrics for KT candidacy assessments and risk stratification. In addition, ESKD-specific obesity management guidelines are needed that will address the neurologic, behavioral, socioeconomic, and physical underpinnings of this increasingly common disease.


The impact of self-rated health on medical care utilization for older people with depressive symptoms.
To test the hypothesis whether self-rated health alone can explain the relationship between depression and medical care utilization for the older people and to determine whether the explanatory power of self-rated health is greater than that of the explanatory power of a major disease and activities of daily living. This study used the data from 1572 older people obtained from the 2005 National Health Interview Survey in Taiwan. The data from the National Health Interview Survey were linked to the 2005 computerized claims data from the National Health Insurance, and from that, the outpatient expenditures and number of outpatient episodes were identified. The contribution of self-rated health, activities of daily living, the presence of major diseases, and self-rated health were estimated using ordinary least squares regressions. Controlling for self-rated health alone almost eliminates the positive relationship between depressive symptoms and number of outpatient visits. After controlling for self-rated health, the utilization ratio of outpatient visits for older people with depressive symptoms reduced significantly to only 1.01 and became insignificant. A similar pattern was observed for total outpatient costs. It was found that self-rated health is an important factor in the depressive symptoms-outpatient utilization relationship. To reduce medical costs for older people with depressive symptoms, it is essential that the self-rated health for this group is improved. Future studies should test the mechanism through which self-rated health impacts on medical utilization for older people with depressive symptoms.


Anatomic and electrophysiologic evaluation of a right lateral atrioventricular Mahaim fiber.
We report a patient who underwent an electrophysiologic study and radiofrequency catheter ablation for a right lateral Mahaim fiber. During sinus rhythm with overt preexcitation, propagation mapping was performed in the right ventricle using a three-dimensional electro-anatomical mapping system (CARTO). Small discrete potentials, which reflected the excitation of the Mahaim fiber, could be recorded along the line from the vicinity of the parental tricuspid annulus to approximately one-third of distal site from the base to the apex. The relationship of the timing of its potential to the anatomical location could be disclosed on recordings of the local electrogram and anatomical map.