Challenges in Permeability Assessment for Oral Drug Product Development.
Drug permeation across the intestinal epithelium is a prerequisite for successful oral drug delivery. The increased interest in oral administration of peptides, as well as poorly soluble and poorly permeable compounds such as drugs for targeted protein degradation, have made permeability a key parameter in oral drug product development. This review describes the various in vitro, in silico and in vivo methodologies that are applied to determine drug permeability in the human gastrointestinal tract and identifies how they are applied in the different stages of drug development. The various methods used to predict, estimate or measure permeability values, ranging from in silico and in vitro methods all the way to studies in animals and humans, are discussed with regard to their advantages, limitations and applications. A special focus is put on novel techniques such as computational approaches, gut-on-chip models and human tissue-based models, where significant progress has been made in the last few years. In addition, the impact of permeability estimations on PK predictions in PBPK modeling, the degree to which excipients can affect drug permeability in clinical studies and the requirements for colonic drug absorption are addressed.


Studies on the particulate lactose synthetase of mouse mammary gland and the role of -lactalbumin in the initiation of lactose synthesis.
1. Some of the kinetic properties of the particulate lactose synthetase of mouse mammary gland were investigated and shown to resemble those that have been reported for the soluble enzyme. Typical values for intact preparations were 2.3mum for the apparent K(m) for alpha-lactalbumin at 40mm-glucose and 1.7mm for the apparent K(m) for glucose at the endogenous concentration of alpha-lactalbumin. 2. Digitonin treatment increased total assayable activity approximately twofold but almost eliminated the endogenous activity found in the absence of alpha-lactalbumin, these findings being consistent with the location of endogenous activity within Golgi vesicles. 3. From the properties of the particulate fraction from lactating mice it was deduced that the effective endogenous alpha-lactalbumin concentrations were in the range 1-10mum. 4. The concentration of alpha-lactalbumin was not significantly different in particles isolated at various stages of pregnancy and early lactation. 5. The implications of these results for the control of lactose synthetase in vivo are discussed.


Casts of chorodial vasculature at physiologic pressures. A new technique.
A method for making latex rubber casts of the ocular vasculature while maintaining physiologic vascular relations has been developed. A series of short-haired domestic ctas, Rhesus monkeys, and albino rabbits were anesthetized and cannulated in the common carotid arteries. The jugular veins were severed, and heparinized saline was infused into the carotid arterial circulation of the animals for complete exsanguination of the head. Throughout the infusion procedure, normal physiologic pressure was maintained. After exsanguination was complete, a solution of latex rubber in distilled water was infused bilaterally and continued until the flow from the jugular veins ceased. The eyes were enucleated and placed in a curing solution. After fixation, the eyes were selectively trimmed and placed into a solution of sodium hydroxide for digestion. The digested tissue was transferred to distilled water for dissection. After dissection, the cast was closely inspected for completeness of capillary filling. Casts such as these are intended for use in observations of ocular vascular anatomy.


Prescribing Pattern of Oral Antihyperglycaemic Drugs, Rationality and Adherence to American Diabetes Association (ADA) Treatment Guidelines among Type 2 Diabetes Mellitus (T2DM) Postmenopausal Women.
Oral antihyperglycaemic prescription trends keep on changing and thus the drug prescription trend study may prove to be powerful exploratory tool for health care providers. To investigate trends in prescriptions of oral antihyperglycaemic drugs (OHDs) among postmenopausal women suffering from T2DM in India and evaluate the rationality and adherence to ADA treatment guidelines. An observational, cross-sectional descriptive prescription audit (n=500) was carried. Postmenopausal women were interviewed in their local language using pre-tested pre validated questionnaire after verbal informed consent at a teaching tertiary care hospital of north India. Oral antihyperglycaemic drugs (OHDs) drugs were categorized as per the pharmacological classification. Adherence to available clinical practice guidelines/recommendations issued under American Diabetes Association (ADA) 2015 Guidelines as well as rationality of these prescriptions were assessed using WHO Guide to Good Prescribing. Mean age of the study population was 58.14±12.86. Mean duration since menopause was 5.3 years and of T2DM was 9.5 years. A 93.4% of the prescriptions had only OHDs whereas 6.6% of the prescriptions had various insulin preprations + OHDs (p<0.0001). Biguanides followed by sulfonylureas, thiazolidinediones, DPP-inhibitors and alpha-glucosidases inhibitor were prescribed in 85.6%, 59.8%, 26.6%, 26% and 12.2% respectively as monotherapy or in combination. Among biguanides, metformin was the most frequently prescribed OHDs. In spite of black box warning on pioglitazone, it was prescribed in 26.6% as FDC. However, clear increase use of vidagliptine was noticed upto 26%. Among combinations most frequent was metformin plus glimipride followed by voglibose plus metformin, whereas, among FDC, metformin plus glimipride followed by metformin plus vidagliptine were most frequently prescribed. Metformin was the most common OHDs to be prescribed followed by glimepiride. Although pioglitazone still continues to be prescribed after safety alert but apparently it appears that the share of pioglitazone has been shifted to vidagliptin or combinations like metformin plus glimipride. Polypharmacy, high use of FDC, & prescription by brand names were some of the irrationalities. Relatively low adherence to ADA treatment guidelines was observed.


Caveolin-1 interacts with the chaperone complex TCP-1 and modulates its protein folding activity.
We report that caveolin-1, one of the major structural protein of caveolae, interacts with TCP-1, a hetero-oligomeric chaperone complex present in all eukaryotic cells that contributes mainly to the folding of actin and tubulin. The caveolin-TCP-1 interaction entails the first 32 amino acids of the N-terminal segment of caveolin. Our data show that caveolin-1 expression is needed for the induction of TCP-1 actin folding function in response to insulin stimulation. Caveolin-1 phosphorylation at tyrosine residue 14 induces the dissociation of caveolin-1 from TCP-1 and activates actin folding. We show that the mechanism by which caveolin-1 modulates TCP-1 activity is indirect and involves the cytoskeleton linker filamin. Filamin is known to bind caveolin-1 and to function as a negative regulator of insulin-mediated signaling. Our data support the notion that the caveolin-filamin interaction contributes to restore insulin-mediated phosphorylation of caveolin, thus allowing the release of active TCP-1.


Locomotor and discriminative stimulus effects of four novel hallucinogens in rodents.
There has been increasing use of novel synthetic hallucinogenic compounds, 2-(4-bromo-2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine hydrochloride (25B-NBOMe), 2-(4-chloro-2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine hydrochloride (25C-NBOMe), 2-(4-iodo-2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine hydrochloride (25I-NBOMe), and N,N-diallyl-5-methoxy tryptamine (5-MeO-DALT), which have been associated with severe toxicities. These four compounds were tested for discriminative stimulus effects similar to a prototypical hallucinogen (-)-2,5-dimethoxy-4-methylamphetamine (DOM) and the entactogen (±)-3,4-methylenedioxymethamphetamine (MDMA). Locomotor activity in mice was tested to obtain dose range and time-course information. 25B-NBOMe, 25C-NBOMe, and 25I-NBOMe decreased locomotor activity. 5-MeO-DALT dose dependently increased locomotor activity, with a peak at 10 mg/kg. A higher dose (25 mg/kg) suppressed activity. 25B-NBOMe fully substituted (≥80%) in both DOM-trained and MDMA-trained rats at 0.5 mg/kg. However, higher doses produced much lower levels of drug-appropriate responding in both DOM-trained and MDMA-trained rats. 25C-NBOMe fully substituted in DOM-trained rats, but produced only 67% drug-appropriate responding in MDMA-trained rats at doses that suppressed responding. 25I-NBOMe produced 74-78% drug-appropriate responding in DOM-trained and MDMA-trained rats at doses that suppressed responding. 5-MeO-DALT fully substituted for DOM, but produced few or no MDMA-like effects. All of the compounds, except 25I-NBOMe, fully substituted for DOM, whereas only 25B-NBOMe fully substituted for MDMA. However, the failure of 25I-NBOMe to fully substitute for either MDMA or DOM was more likely because of its substantial rate-depressant effects than weak discriminative stimulus effects. All of the compounds are likely to attract recreational users for their hallucinogenic properties, but probably of much less interest as substitutes for MDMA. Although no acute adverse effects were observed at the doses tested, the substantial toxicities reported in humans, coupled with the high likelihood for illicit use, suggests that these compounds have the same potential for abuse as other, currently scheduled compounds.


Longitudinal Trajectories of Children's Physical Activity and Sedentary Behaviors on Weekdays and Weekends.
The objective was to examine changes of children's time spent in sedentary, light physical activity, moderate to vigorous physical activity (MVPA), and estimated energy expenditure (EE) rates during weekdays and weekends across 3 years. An initial sample of 261 children's (mean age = 7.81 y) 5-day physical activity and EE were assessed annually via accelerometry across 3 years using repeated-measures multivariate analysis of variance. The outcome variables were time spent in sedentary, light physical activity, MVPA, and kilocalories per day for weekdays and weekends. A significant decrease in MVPA and EE occurred during weekdays across the 3 years (P = .01). Only the second-year data demonstrated an increase (+2.49 min) in weekend MVPA (P = .04). Children's sedentary time during weekdays increased significantly in years 1 and 2 (P = .01), yet significantly decreased in the third year (-44.31 min). Children's sedentary time during weekends significantly decreased in the first year (-27.31 min), but increased in the following 2 years (P = .01). Children's light physical activity demonstrated a statistically significant increase in year 2 (+3.75 min) during weekdays (P = .05). Children's MVPA and EE generally declined during weekdays but were maintained during weekends across a 3-year time span. Children may benefit most from weekday intervention strategies.


Aerobic Exercise During Encoding Impairs Hippocampus-Dependent Memory.
We investigated how aerobic exercise during encoding affects hippocampus-dependent memory through a source memory task that assessed hippocampus-independent familiarity and hippocampus-dependent recollection processes. Using a within-participants design, young adult participants performed a memory-encoding task while performing a cycling exercise or being seated. The subsequent retrieval phase was conducted while sitting on a chair. We assessed behavioral and event-related brain potential measures of familiarity and recollection processes during the retrieval phase. Results indicated that source accuracy was lower for encoding with exercise than for encoding in the resting condition. Event-related brain potential measures indicated that the parietal old/new effect, which has been linked to recollection processing, was observed in the exercise condition, whereas it was absent in the rest condition, which is indicative of exercise-induced hippocampal activation. These findings suggest that aerobic exercise during encoding impairs hippocampus-dependent memory, which may be attributed to inefficient source encoding during aerobic exercise.


Prominent expression of xenobiotic efflux transporters in mouse extraembryonic fetal membranes compared with placenta.
Fetal exposure to xenobiotics can be restricted by transporters at the interface between maternal and fetal circulation. Previous work identified transporters in the placenta; however, less is known about the presence of these transporters in the fetal membranes (i.e., yolk sac and amniotic membranes). The purpose of this study was to quantify mRNA and protein expression of xenobiotic transporters in mouse placenta and fetal membranes during mid to late gestation. Concepti (placenta and fetal membranes, gestation day 11) or placenta and fetal membranes (gestation days 14 and 17) were collected from pregnant mice and analyzed for expression of multidrug resistance-associated proteins (Mrps), multidrug resistance proteins (Mdrs), multidrug and toxin extrusion proteins (Mates), breast cancer resistance protein (Bcrp), and organic anion-transporting polypeptides (Oatps). Maternal liver and kidneys were also collected at day 14 for mRNA and immunohistochemical analysis. mRNA expression of Mrp, Mdr, Bcrp, Mate-1, and Oatp isoforms was detected at day 11. The uptake carriers Oatp2a1, 3a1, 4a1, and 5a1 showed placenta-predominant expression. At days 14 and 17, fetal membranes expressed higher mRNA levels of the efflux transporters Mrp2 (7-fold), Mrp4 (5-fold), Mrp5 (3-fold), Mrp6 (12-fold), Bcrp (2-fold), and Mate-1 (7-fold) than placenta. Western blot analysis of Mrp2, Mrp4, Mrp6, and Bcrp confirmed higher expression in fetal membranes. Immunostaining revealed apical (Mrp2 and Bcrp) and basolateral (Mrp4, 5, and 6) cellular localization in epithelial cells of the yolk sac. In conclusion, xenobiotic transporters in the fetal membranes may provide an additional route to protect the fetus against endogenous chemicals and xenobiotics.


Beta interferon controls West Nile virus infection and pathogenesis in mice.
Studies with mice lacking the common plasma membrane receptor for type I interferon (IFN-αβR(-)(/)(-)) have revealed that IFN signaling restricts tropism, dissemination, and lethality after infection with West Nile virus (WNV) or several other pathogenic viruses. However, the specific functions of individual IFN subtypes remain uncertain. Here, using IFN-β(-)(/)(-) mice, we defined the antiviral and immunomodulatory function of this IFN subtype in restricting viral infection. IFN-β(-)(/)(-) mice were more vulnerable to WNV infection than wild-type mice, succumbing more quickly and with greater overall mortality, although the phenotype was less severe than that of IFN-αβR(-)(/)(-) mice. The increased susceptibility of IFN-β(-)(/)(-) mice was accompanied by enhanced viral replication in different tissues. Consistent with a direct role for IFN-β in control of WNV replication, viral titers in ex vivo cultures of macrophages, dendritic cells, fibroblasts, and cerebellar granule cell neurons, but not cortical neurons, from IFN-β(-)(/)(-) mice were greater than in wild-type cells. Although detailed immunological analysis revealed no major deficits in the quality or quantity of WNV-specific antibodies or CD8(+) T cells, we observed an altered CD4(+) CD25(+) FoxP3(+) regulatory T cell response, with greater numbers after infection. Collectively, these results suggest that IFN-β controls WNV pathogenesis by restricting infection in key cell types and by modulating T cell regulatory networks.


Three-year outcomes of a once daily fractionation scheme for accelerated partial breast irradiation (APBI) using 3-D conformal radiotherapy (3D-CRT).
The aim of this study was to report 3-year outcomes of toxicity, cosmesis, and local control using a once daily fractionation scheme (49.95 Gy in 3.33 Gy once daily fractions) for accelerated partial breast irradiation (APBI) using three-dimensional conformal radiotherapy (3D-CRT). Between July 2008 and August 2010, women aged ≥40 years with ductal carcinoma in situ or node-negative invasive breast cancer ≤3 cm in diameter, treated with breast-conserving surgery achieving negative margins, were accrued to a prospective study. Women were treated with APBI using 3-5 photon beams, delivering 49.95 Gy over 15 once daily fractions over 3 weeks. Patients were assessed for toxicities, cosmesis, and local control rates before APBI and at specified time points. Thirty-four patients (mean age 60 years) with Tis 0 (n = 9) and T1N0 (n = 25) breast cancer were treated and followed up for an average of 39 months. Only 3% (1/34) patients experienced a grade 3 subcutaneous fibrosis and breast edema and 97% of the patients had good/excellent cosmetic outcome at 3 years. The 3-year rate of ipsilateral breast tumor recurrence (IBTR) was 0% while the rate of contralateral breast events was 6%. The 3-year disease-free survival (DFS), overall survival (OS), and breast cancer-specific survival (BCSS) was 94%, 100%, and 100%, respectively. Our novel accelerated partial breast fractionation scheme of 15 once daily fractions of 3.33 Gy (49.95 Gy total) is a remarkably well-tolerated regimen of 3D-CRT-based APBI. A larger cohort of patients is needed to further ascertain the toxicity of this accelerated partial breast regimen.


Dependence of initiation of an innocent bystander cytotoxicity reaction on the association of viral antigens and tumor cell membranes in mice.
In chemically induced mouse mammary tumors in BALB/c mice, no murine mammary tumor virus (MuMTV) antigens could be detected in the tumor cell membranes. In contrast, in mammary tumors of spontaneous appearance in BALB/cfC3H mice neonatally infected with MuMTV, viral antigens could be detected by immunofluorescence. Specific activation of immune spleen lymphocytes in vitro by homologous tumor cell membrane preparations resulted in an innocent bystander cytotoxicity reaction in BALB/c mice bearing either the chemical- or virus-induced mammary tumors. Non-tumor bearers did not respond in this reaction. The cytotoxicity effector cell was a nylon-adherent Thy 1.2+, Lyt 1+, Lyt 2+ lymphocyte. Induction of the reaction in the chemically induced mammary tumor bearers was related to tumor-associated antigens, but in the mice with MuMTV-induced tumors, it was possible that all responses were solely due to viral antigens. Biochemical analyses using immunoprecipitation techniques indicated that the major external glycoprotein of MuMTV (gp52) was present in the tumor cell membrane preparations. Preincubation of the cell membranes of virus-induced tumors with anti-MuMTV completely blocked the capacity of this preparation to induce the cytotoxicity. Preabsorption of the anti-MuMTV with purified MuMTV removed all the blocking activity of these sera, indicating that the antigenic determinants recognized were of viral origin. However, purified MuMTV failed by itself to elicit the innocent bystander cytotoxicity. This observation indicates that an association with membranes was necessary for the viral antigens to initiate and/or effect this cell-mediated immune reaction.


An investigation by e.p.r. and optical spectroscopy of cytochrome oxidase during turnover.
Cytochrome oxidase (EC 18.104.22.168; ferrocytochrome c:oxygen oxidoreductase) was studied during steady-state by optical and e.p.r. methods. Starting with either the 'resting' or the 'pulsed' enzyme, oxidase, cytochrome c, ascorbate and O2 were mixed and the reaction monitored optically. Tetramethylphenylenediamine was used as mediator to poise the steady-state to the desired reduction level. After mixing, the reaction was quenched by the used of rapid-freeze techniques. The e.p.r. spectra of samples captured at increasing tetramethylphenylenediamine concentrations (i.e. higher electron flux) show decreasing g = 2 (Cu A) and g = 3 (cytochrome a) signals. No Cu B or g = 6 signals (high-spin cytochrome a3) could be found during the reaction. Also, the signal with peaks at g = 1.69, 1.78 and 5 as well as the g = 12 signal was hardly detectable at higher turnover rates. The only new signal appearing during turnover is a radical signal, which is discussed in terms of a protein radical. Finally, a scheme is presented, proposing a catalytic cycle for cytochrome oxidase with respect to the O2 binding Cu B-cytochrome a3 unit.


Sexual and other axis I diagnoses of 60 males arrested for crimes against children involving the Internet.
This study was conducted to describe Axis I sexual diagnoses of 60 males arrested for possession of child pornography obtained via the Internet and/or attempting to meet children via the Internet. Data was obtained from a chart review of evaluations conducted on 60 males referred for a psychosexual evaluation following an arrest for possession of child pornography and/or attempting to meet children. All crimes involved use of the Internet. Information obtained from the chart review was entered into SAS. All diagnoses were made according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision. Simple descriptive statistics were computed and cross tabulations were tested for significance using chi2 or Fisher's Exact test. Of the total sample, 40% had at least one paraphilia. Thirty-one percent had a diagnosis of pedophilia and 18% of a paraphilia not otherwise specified (NOS). Thirty-three percent had a sexual disorder NOS, characterized by hypersexuality. Seventy percent of the total sample had an Axis I disorder that antedated and was judged to be contributory to the behavior leading to their arrest. This sample of men arrested for committing crimes against children and adolescents via the Internet has a high incidence of lifetime sexual and other psychopathology.


A mammalian microRNA cluster controls DNA methylation and telomere recombination via Rbl2-dependent regulation of DNA methyltransferases.
Dicer initiates RNA interference by generating small RNAs involved in various silencing pathways. Dicer participates in centromeric silencing, but its role in the epigenetic regulation of other chromatin domains has not been explored. Here we show that Dicer1 deficiency in Mus musculus leads to decreased DNA methylation, concomitant with increased telomere recombination and telomere elongation. These DNA-methylation defects correlate with decreased expression of Dnmt1, Dnmt3a and Dnmt3b DNA methyltransferases (Dnmts), and methylation levels can be recovered by their overexpression. We identify the retinoblastoma-like 2 protein (Rbl2) as responsible for decreased Dnmt expression in Dicer1-null cells, suggesting the existence of Dicer-dependent small RNAs that target Rbl2. We identify the miR-290 cluster as being downregulated in Dicer1-deficient cells and show that it silences Rbl2, thereby controlling Dnmt expression. These results identify a pathway by which miR-290 directly regulates Rbl2-dependent Dnmt expression, indirectly affecting telomere-length homeostasis.


Controlling Salmonella: strategies for feed, the farm, and the processing plant.
Controlling Salmonella in poultry is an ongoing food safety measure and while significant progress has been made, there is a need to continue to evaluate different strategies that include understanding Salmonella-poultry interaction, Salmonella-microbiota interactions, Salmonella genetics and response to adverse conditions, and preharvest and postharvest parameters that enable persistence. The purpose of this symposium is to discuss different strategies to consider from feed milling to the farm to the processing environment. This Poultry Science Association symposium paper is divided into 5 different sections that covers 1) immunological aspects of Salmonella control, 2) application of Salmonella genetics for targeted control strategies in poultry production, 3) improving poultry feed hygienics: utilizing feed manufacture techniques and equipment to improve feed hygienics, 4) practical on farm interventions for controlling Salmonella-what works and what may not work, and 5) monitoring and mitigating Salmonella in poultry. These topics elucidate the critical need to establish control strategies that will improve poultry gut health and limit conditions that exposes Salmonella to stress causing alterations to virulence and pathogenicity both at preharvest and postharvest poultry production. This information is relevant to the poultry industry's continued efforts to ensure food safety poultry production.


Indirect interactions influence contact network structure and diffusion dynamics.
Interaction patterns at the individual level influence the behaviour of diffusion over contact networks. Most of the current diffusion models only consider direct interactions, capable of transferring infectious items among individuals, to build transmission networks of diffusion. However, delayed indirect interactions, where a susceptible individual interacts with infectious items after the infected individual has left the interaction space, can also cause transmission events. We define a diffusion model called the same place different time transmission (SPDT)-based diffusion that considers transmission links for these indirect interactions. Our SPDT model changes the network dynamics where the connectivity among individuals varies with the decay rates of link infectivity. We investigate SPDT diffusion behaviours by simulating airborne disease spreading on data-driven contact networks. The SPDT model significantly increases diffusion dynamics with a high rate of disease transmission. By making the underlying connectivity denser and stronger due to the inclusion of indirect transmissions, SPDT models are more realistic than same place same time transmission (SPST)-based models for the study of various airborne disease outbreaks. Importantly, we also find that the diffusion dynamics including indirect links are not reproducible by the current SPST models based on direct links, even if both SPDT and SPST networks assume the same underlying connectivity. This is because the transmission dynamics of indirect links are different from those of direct links. These outcomes highlight the importance of the indirect links for predicting outbreaks of airborne diseases.


Liquid crystal codendrimers with a statistical distribution of phenolic and mesogenic groups: behavior as Langmuir and Langmuir-Blodgett films.
The first series of carbosilane liquid crystal codendrimers with groups of different polarity has been synthesized. The chemical structure of the newly synthesized materials and the composition of the codendrimers were studied by NMR spectroscopy and MALDI-TOF MS. It was found that the codendrimers tend to form stable Langmuir films at the air-water surface. The influence of composition and generation number on surface pressure-surface area isotherms and film stability was studied. Brewster angle microscopy confirmed the different phase behavior for monolayers of different codendrimer composition and generation number. It was found that side groups of fifth-generation codendrimers do not segregate, unlike those of lower generations. Langmuir-Blodgett films on solid substrates were obtained by the vertical dipping method. X-ray diffraction showed that the codendrimers with 75% of hydrophobic mesogenic terminal groups formed ordered layers parallel to the substrate.


Arterial stiffness is not increased in patients with short duration rheumatoid arthritis and ankylosing spondylitis.
Patients with rheumatoid arthritis (RA) and ankylosing spondylitis (AS) have increased cardiovascular (CV) morbidity and mortality. Arterial stiffness is an independent predictor of CV events. The aim of the study was to assess arterial stiffness and inflammatory markers in patients with short duration chronic arthritis. We assessed carotid-femoral pulse wave velocity (PWV), augmentation index (AIx), traditional CV risk factors and inflammatory and endothelial markers in 71 chronic arthritis patients (RA and AS) and in 29 healthy controls. We did not find differences in PWV (for RA, AS and controls, respectively: 10 [8.8-10.9] versus 10.7 [9.1-11.8] versus 9.2 [8.3-11.4] m/s; p = .14) and AIx (for RA, AS and controls, respectively: 24.3 ± 11.5 versus 5.7 ± 12.4 versus 10 ± 12.8%; p = .22). Both groups of arthritis patients had active disease with significantly elevated inflammatory markers compared to controls. There were no correlations between endothelial and inflammatory markers and parameters of arterial stiffness in arthritis patients. When analyzing arthritis patients according to median of PVW, there were no significant differences in inflammatory and endothelial markers. We found that in patients with short duration active RA and AS arterial stiffness was not increased and furthermore, there was no association between markers of systemic inflammation and arterial stiffness.


Experience in snuffbox arteriovenous fistulae for hemodialysis.
Sixty-five patients with "Snuffbox" arteriovenous fistulae for hemodialysis were reviewed. The procedure was performed under local anaesthesia as an outpatient procedure. It was done as a "standby" procedure for 41.5% of patients with Chronic Ambulatory Peritoneal Dialysis. 58.5% of patients had the procedure done for primary hemodialysis. Patency was assessed as presence of an engorged vein and presence of thrill. This was assessed routinely at 4 to 6 weeks after the procedure. Patency rate was noted to be 83%. The commonest complication was thrombosis of the arteriovenous anastomosis. Some patients required repeated procedure at a more proximal site. Further study to determine the blood flow rate provided by the anastomosis, and comparison with other anatomical sites is necessary.


The synthesis of inositol hexakisphosphate. Characterization of human inositol 1,3,4,5,6-pentakisphosphate 2-kinase.
The enzyme(s) responsible for the production of inositol hexakisphosphate (InsP(6)) in vertebrate cells are unknown. In fungal cells, a 2-kinase designated Ipk1 is responsible for synthesis of InsP(6) by phosphorylation of inositol 1,3,4,5,6-pentakisphosphate (InsP(5)). Based on limited conserved sequence motifs among five Ipk1 proteins from different fungal species, we have identified a human genomic DNA sequence on chromosome 9 that encodes human inositol 1,3,4,5,6-pentakisphosphate 2-kinase (InsP(5) 2-kinase). Recombinant human enzyme was produced in Sf21 cells, purified, and shown to catalyze the synthesis of InsP(6) or phytic acid in vitro. The recombinant protein converted 31 nmol of InsP(5) to InsP(6)/min/mg of protein (V(max)). The Michaelis-Menten constant for InsP(5) was 0.4 microM and for ATP was 21 microM. Saccharomyces cerevisiae lacking IPK1 do not produce InsP(6) and show lethality in combination with a gle1 mutant allele. Here we show that expression of the human InsP(5) 2-kinase in a yeast ipk1 null strain restored the synthesis of InsP(6) and rescued the gle1-2 ipk1-4 lethal phenotype. Northern analysis on human tissues showed expression of the human InsP(5) 2-kinase mRNA predominantly in brain, heart, placenta, and testis. The isolation of the gene responsible for InsP(6) synthesis in mammalian cells will allow for further studies of the InsP(6) signaling functions.


The lncRNA BCYRN1 Functions as an Oncogene in Human Glioma by Downregulating miR-125a-5p in vitro.
Numerous studies have demonstrated that long noncoding RNAs (lncRNAs) are deregulated in many cancers and exert their functions through multiple cancer-related biological processes. Glioma is the most common primary malignant central nervous system tumor and has a high fatality rate in adults. In current study, we aimed to determine the role and functional mechanism of the lncRNA BCYRN1 in glioma. Gain-of-function and loss-of function approaches were used to investigate the function of BCYRN1. The effects of BCYRN1 on glioma cell proliferation, migration and invasion were evaluated using MTS, Transwell and wound-healing assays. The correlation between the expression of BCYRN1 and miR-125a-5p was verified by quantitative real-time PCR. The upregulation of BCYRN1 promoted the proliferation, migration and invasion of glioma cells. Meanwhile, the knockdown of BCYRN1 had the opposite effects. BCYRN1 was negatively correlated with miR-125a-5p. Additionally, TAZ, the endogenous target of miR-125a-5p, could be regulated by BCYRN1 in RNA and protein levels. A miR-125a-5p inhibitor restored BCYRN1 siRNA function in glioma. The present study indicates that BCYRN1 promotes glioma cell proliferation, invasion and migration in vitro. Mechanistically, upregulated expression of BCYRN1 in glioma acts as a sponge to sequester the endogenous tumor suppressor miR-125a-5p and to further increase the expression TAZ. Our findings suggest that BCYRN1 is a novel oncogene and a new therapeutic target for glioma.


Review of collagen I hydrogels for bioengineered tissue microenvironments: characterization of mechanics, structure, and transport.
Type I collagen hydrogels have been used successfully as three-dimensional substrates for cell culture and have shown promise as scaffolds for engineered tissues and tumors. A critical step in the development of collagen hydrogels as viable tissue mimics is quantitative characterization of hydrogel properties and their correlation with fabrication parameters, which enables hydrogels to be tuned to match specific tissues or fulfill engineering requirements. A significant body of work has been devoted to characterization of collagen I hydrogels; however, due to the breadth of materials and techniques used for characterization, published data are often disjoint and hence their utility to the community is reduced. This review aims to determine the parameter space covered by existing data and identify key gaps in the literature so that future characterization and use of collagen I hydrogels for research can be most efficiently conducted. This review is divided into three sections: (1) relevant fabrication parameters are introduced and several of the most popular methods of controlling and regulating them are described, (2) hydrogel properties most relevant for tissue engineering are presented and discussed along with their characterization techniques, (3) the state of collagen I hydrogel characterization is recapitulated and future directions are proposed. Ultimately, this review can serve as a resource for selection of fabrication parameters and material characterization methodologies in order to increase the usefulness of future collagen-hydrogel-based characterization studies and tissue engineering experiments.


The frequency of CYP2C9, VKORC1, and CYP4F2 polymorphisms in Russian patients with high thrombotic risk.
VKORC1, CYP2C9, and CYP4F2 are known to be responsible for the metabolism of warfarin. The aim was to explore the frequencies of these genotypes in the Russian population and compare the results with those for other populations. In total, 91 Caucasian subjects with a mean age of 66.17 years (SD, 10.9) were recruited into the study. Of them, 40 patients (48.2%) were men. In order to obtain necessary clinical data, the medical records of the patients were reviewed. Blood (5 mL) was taken from each subject, and DNA was isolated and used for identification of the CYP2C9 allele *1, *2, *3, -1639G/A VKORC1, and CYP4F2 V433M rs2108622 C>T, using the real-time polymerase chain reaction-restriction fragment length polymorphism assay. The CYP2C9*1/*1 genotype was detected in 67.0%, CYP2C9*1/*2 in 9.9%, CYP2C9*1/*3 in 11.0%, CYP2C9*2/*2 in 2.2%, CYP2C9*2/*3 in 8.8%, and CYP2C9*3/*3 in 1.1% of the patients. The results for VKORC1 were as follows: 49.5% (GG), 28.6% (GA), and 22.0% (AA); meanwhile, those for the genotype CYP4F2 were 57.1% (CC), 34.1% (CT), and 7.7% (TT). No significant deviations from the Hardy-Weinberg equilibrium were observed. The frequency of the polymorphisms in the Russian population was found to differ from Asian and close to Caucasian. There were no significant interethnic variations in the frequency of CYP4F2 among Russian, Asian, and Caucasian populations. The frequency of CYP2C9, CYP4F2, and VKORC1 polymorphisms in Russian patients is comparable with other European ethnic groups.


Physical activity behavior in older women with rheumatoid arthritis.
To identify physical activity behaviors of older (>/=60 years) women with rheumatoid arthritis, and to determine if physical activity behavior differed by demographic characteristics or disease duration. Cognitively intact, ambulatory women with rheumatoid arthritis seen at an urban university arthritis center (n = 185) responded to a survey that included demographics, number of years since RA diagnosis, and physical activity assessed with the Yale Physical Activity Survey. Participants had a mean age of 70 years and a mean disease duration of 17.6 years. Mean total of physical activity, including low-, moderate-, and high-intensity activity, was 23 hours/week, with 47 different physical activities identified. Housework comprised 67% of total physical activity time, whereas leisure activities and planned exercise comprised only 15% and 10%, respectively. Sixty percent of the women reported participation in some type of vigorous activity, and 88% reported they had done leisure walking in the past month. Age was negatively associated with scores on the activity summary index (r = -0.195, P < 0.01), and there was a significant difference on the activity summary index by employment status, with women in the work force (n = 47) and those involved in housekeeping (n = 105) scoring significantly higher (P = 0.003) than the women (n = 33) who described themselves as retired (F = 7.81, 2 degrees of freedom, P = 0.001). Older women with rheumatoid arthritis may participate in a broader array of physical activities than previously assumed, and could benefit from increasing the proportion of moderate or vigorous activity incorporated into their daily routines.


Simulated Microgravity Changes the Number of Mechanically Gated and Mechanosensitive Ion Channels Genes Transcripts in Rat Ventricular Cardiomyocytes.
The mechanoelectrical feedback in the heart is based on the work of mechanically gated (MGCs) and mechanosensitive (MSCs) channels. Since microgravity alters the heart's morphological and physiological properties, we hypothesized that the expression of both MGCs and MSCs would be affected. We employed RNA transcriptome sequencing to investigate changes in the gene transcript levels of MGCs and MSCs in isolated rat ventricular cardiomyocytes under control conditions and in a simulated microgravity environment. For the first time, our findings demonstrated that simulated microgravity induces alterations in the gene transcript levels of specific MGCs, such as TRPM7, TRPV2, TRPP1, TRPP2, Piezo1, TMEM63A, TMEM36B, and known MSCs, including K2P2.1, K2P3.1, Kir6.1, Kir6.2, NaV1.5, CaV1.2, KV7.1. However, other voltage-gated channels and channels lacking a voltage sensor remained unaffected. These findings suggest that the altered expression of MGCs and MSCs could lead to changes in the net currents across the membrane, ultimately impacting the heart's function.


[Yellow nail syndrome: case report].
The yellow nail syndrome is a rare disease, in which there is a triad of lymphedema, pleural effusion and slow-growing dystrophic yellow nails. Many associations have already been described; among them, chronic respiratory tract diseases, autoimmune disorders, malignancies and immunodeficiency conditions. Only one third of cases in the literature show all findings. The case reported next is an example of the classical triad.


Texture Parameters Measured by UHF-MRI and CT Scan Provide Information on Bone Quality in Addition to BMD: A Biomechanical Ex Vivo Study.
The current definition of osteoporosis includes alteration of bone quality. The assessment of bone quality is improved by the development of new texture analysis softwares. Our objectives were to assess if proximal femoral trabecular bone texture measured in Ultra high field (UHF) 7 Tesla MRI and CT scan were related to biomechanical parameters, and if the combination of texture parameters and areal bone mineral density (aBMD) measured by dual-energy X-ray absorptiometry provided a better prediction of femoral failure than aBMD alone. The aBMD of 16 proximal femur ends from eight cadavers were investigated. Nineteen textural parameters were computed in three regions or volumes of interest for each specimen on UHF MRI and CT scan. Then, the corresponding failure load and failure stress were calculated thanks to mechanical compression test. aBMD was not correlated to failure load (R2 = 0.206) and stress (R2 = 0.153). The failure load was significantly correlated with ten parameters in the greater trochanter using UHF MRI, and with one parameter in the neck and the greater trochanter using CT scan. Eight parameters in the greater trochanter using UHF MRI combined with aBMD improved the failure load prediction, and seven parameters improved the failure stress prediction. Our results suggest that textural parameters provide additional information on the fracture risk of the proximal femur when aBMD is not contributive.


Hepatic fibrosis and its serum markers: a review.
Hepatic fibrosis is characterized by a progressive increase in extracellular matrix in the liver, formed by collagens, proteoglycans and glycoproteins, qualitatively similar but quantitatively different from that in normal liver. A great number of matrix-related substances have been investigated in serum in order to identify reliable serum markers of liver fibroplasia. Among the various cleavage products of collagen precursor, the NPIIIP collagen is at present considered the most reliable serum marker of active fibrogenesis in liver, useful in monitoring the progression of fibrosis and in assessing the therapeutic efficacy of antifibrotic drugs. Lam-P1 and type IV collagen are now regarded as putative markers of basement membrane formation and sinusoids capillarization, an important pathological process in fibrosing disease, related to the impairment of hepatic circulation. Other serum-measured matrix-related substances, e.g. enzymes involved in collagen metabolism, fibronectin and proteoglycans, have not proved to reflect liver fibroplasia reliably. In spite of the availability of useful serum markers, the assessment of hepatic fibrosis is still based on liver biopsy.


Effects of aging on hemispheric asymmetry in inferior frontal cortex activity during belief-bias syllogistic reasoning: a near-infrared spectroscopy study.
The belief-bias effect in syllogistic reasoning refers to the tendency for subjects to be erroneously biased when logical conclusions are incongruent with beliefs about the world. This study examined age-related differences in inferior frontal cortex (IFC) activity associated with belief-bias reasoning using near-infrared spectroscopy (NIRS). The subjects were 32 older (mean age, 68.53 years) and 32 young adult volunteers (mean age, 21.50 years). They performed belief-congruent and incongruent reasoning trials while right and left IFC activities were being measured by NIRS. Behavioral analysis found that older adults exhibited a larger belief-bias than young adults. NIRS analysis showed that the right IFC was more activated than the left IFC in young adults, while there was no significant hemispheric difference in older adults. On correlation analysis, there was a significant positive correlation between reasoning accuracy and IFC activation in both hemispheres for older adults, while in young adults, the correlation was significant only in the right hemisphere. These correlation patterns suggest that the right IFC is critical for resolving conflicting reasoning in young adults, but that older adults may further recruit the left IFC to compensate for the age-related decline in the inhibitory control functions. Thus, we demonstrate, for the first time, age-related differences in neural activity associated with belief-bias reasoning.


Monitoring access following Medicare price changes: physician perspective.
In this article, the author examines changes in Medicare beneficiaries' access to services following the Omnibus Budget Reconciliation Act of 1987 "overpriced" procedure price reductions from the physician perspective. Three measures of physician availability remained essentially constant: number of physicians treating beneficiaries or performing overpriced procedures; average Medicare caseload; and average share of a physician's Medicare practice comprised of those who are poor and not white. Physician practice characteristics were examined and provided evidence of continuing participation in Medicare: Average Medicare revenue increased 10 percent, and average volume of all services increased. However, physicians with the largest fee reductions or who were the most financially dependent on the procedures did not change overpriced procedure volume.


Agrin binds alpha-synuclein and modulates alpha-synuclein fibrillation.
Recent studies have begun to investigate the role of agrin in brain and suggest that agrin's function likely extends beyond that of a synaptogenic protein. Particularly, it has been shown that agrin is associated with the pathological lesions of Alzheimer's disease (AD) and may contribute to the formation of beta-amyloid (Abeta) plaques in AD. We have extended the analysis of agrin's function in neurodegenerative diseases to investigate its role in Parkinson's disease (PD). Alpha-synuclein is a critical molecular determinant in familial and sporadic PD, with the formation of alpha-synuclein fibrils being enhanced by sulfated macromolecules. In the studies reported here, we show that agrin binds to alpha-synuclein in a heparan sulfate-dependent (HS-dependent) manner, induces conformational changes in this protein characterized by beta-sheet structure, and enhances insolubility of alpha-synuclein. We also show that agrin accelerates the formation of protofibrils by alpha-synuclein and decreases the half-time of fibril formation. The association of agrin with PD lesions was also explored in PD human brain, and these studies shown that agrin colocalizes with alpha-synuclein in neuronal Lewy bodies in the substantia nigra of PD brain. These studies indicate that agrin is capable of accelerating the formation of insoluble protein fibrils in a second common neurodegenerative disease. These findings may indicate shared molecular mechanisms leading to the pathophysiology in these two neurodegenerative disorders.


Outcome of Proximal Femoral Nail Antirotation II fixation of Peritrochanteric Fracture of Femur.
Intramedullary fixation is biomechanically superior to extramedullary fixation for fracture of peritrochanteric region of femur in elderly which is often complicated due to associated osteoporosis. Helical blade of proximal femoral nail antirotation II has stronger purchase in the head of femur preventing rotation and cutout. This study was done to evaluate the outcome of Proximal Femoral Nail Antirotation II fixation in B.P. Koirala Institute of Health Sciences. Retrospective review of the data of 100 traumatic peritrochanteric fracture cases operated between March 2017 and March 2018 was done for study. Cases lost to follow-up, deaths and incomplete functional outcome (VAS, HHS), intraoperative and postoperative data were excluded. Total of 71 cases were included in the study. Mean age of patients was 65 (+14) (range:19 to 86) years and were operated for intertrochanteric (54), subtrochanteric (16) and neck of femur (1) fractures. Trivial fall on ground was the commonest mechanism of injury (43 of 71). Mean injury to surgery time was 7 (+7) days and mean duration of hospital stay was 5.92 (+4) days. Mean blood loss during surgery was 132.61 (+21) ml and mean surgical time was 56 (+8) minutes. Mean visual analogue scale (VAS) Score was 7 at 2 weeks, 3 at 6 weeks and 1 at 12 weeks. Mean harris hip score was 80 (+10) at 6 months. Proximal femoral nail antirotation II is a reliable implant system to use for fixation of proximal femoral fractures with a good functional outcome and low complication rates.


Effect of Tinnitus Habituation Therapy on Auditory Abilities.
Introduction Research with tinnitus patients presenting with hearing loss and normal hearing subjects have verified that tinnitus interferes with auditory processing. One treatment option for this symptom consists of tinnitus habituation therapy. Objective To determine the influence of tinnitus habituation therapy on auditory abilities, on the self-perception of tinnitus regarding loudness and discomfort, and its impact on the quality of life. Methods A quasi-experimental study was conducted with 19 individuals with tinnitus. Audiological tinnitus and auditory processing evaluations were performed. Subsequently, the volunteers underwent tinnitus habituation therapy for 6 months. Ultimately, all of the individuals were reevaluated. Results A statistically significant reduction in tinnitus self-perception was observed in relation to loudness, discomfort, and its impact on the quality of life ( p = 0.001, 0.001, 0.023, respectively). However, the influence of tinnitus habituation therapy on auditory abilities was not verified. Conclusion Tinnitus habituation therapy was effective in reducing the self-perception of tinnitus loudness and discomfort, as well as the impact of the symptom on the quality of life. However, these factors did not improve the auditory processing skills in tinnitus patients.


In vitro reconstruction of branched tubular structures from lung epithelial cells in high cell concentration gradient environment.
We have succeeded in developing hollow branching structure in vitro commonly observed in lung airway using primary lung airway epithelial cells. Cell concentration gradient is the key factor that determines production of the branching cellular structures, as optimization of this component removes the need for heterotypic culture. The higher cell concentration leads to the more production of morphogens and increases the growth rate of cells. However, homogeneous high cell concentration does not make a branching structure. Branching requires sufficient space in which cells can grow from a high concentration toward a low concentration. Simulation performed using a reaction-diffusion model revealed that long-range inhibition prevents cells from branching when they are homogeneously spread in culture environments, while short-range activation from neighboring cells leads to positive feedback. Thus, a high cell concentration gradient is required to make branching structures. Spatial distributions of morphogens, such as BMP-4, play important roles in the pattern formation. This simple yet robust system provides an optimal platform for the further study and understanding of branching mechanisms in the lung airway, and will facilitate chemical and genetic studies of lung morphogenesis programs.


The effectiveness of flexible ureterorenoscopy for opaque and non-opaque renal stones.
To evaluate the treatment success rate of flexible ureterorenoscopy (URS) for opaque and non-opaque renal stones. Ninety-four patients, who underwent flexible URS for renal stones between October 2012 and January 2014, were included. The patients were divided into two groups according to stone radiolucency. The patients were evaluated with preoperative and postoperative (at the weeks 4) computed tomography. Success of the treatment was defined as stone-free status and residual fragments < 4 mm. Success of the treatment was observed in 79 (84%) patients. Sex, stone size, and stone location were factors affecting treatment success. Seventy-five (79.8%) patients had opaque stones, and 19 (20.2%) had non-opaque stones. The treatment success rates for opaque and non-opaque stones were 86.6% and 73.6%, respectively (P = .167). Flexible URS was a successful modality with acceptable morbidity to treat renal stones. These results show that radiolucent and opaque stones can be effectively treated by flexible URS.


DNA based multi-copper ions assembly using combined pyrazole and salen ligandosides.
The DNA structure is an ideal building block for the construction of functional nano-objects. In this direction, metal coordinating base pairs (ligandosides) are an appealing tool for the future specific functionalization of such nano-objects. We present here a study, in which we combine the metal ion coordinating pyrazole ligandoside with the interstrand crosslinking salen ligandoside system. We show that both ligandosides, when combined, are able to create stable multi-copper ion complexing DNA double helix structures in a cooperative fashion.


Case Report: Hemorrhagic Fever with Renal Syndrome Complicated by Bilateral Subdural Hematoma.
Hemorrhagic fever with renal syndrome (HFRS) is an acute, natural focal disease worldwide. Bilateral subdural hematoma (BSH) is a rare occurrence in patients with HFRS. A 51-year-old man was admitted with fever, headache, lower back pain, and reduced urine volume. The patient was diagnosed with HFRS accompanied by BSH, as evidenced by IgM and IgG antibodies for hantavirus that were positive, and abnormal blood test results and computed tomographic head scan. He recovered and was discharged after symptomatic treatment. Hemorrhagic fever with renal syndrome might present rare clinical manifestations with BSH. The early identification of this condition is crucial to an improved prognosis.


Age-dependent interaction between Life's Essential 8 and chronic kidney disease: A national cross-sectional analysis.
Chronic kidney disease (CKD), an age-related condition, is closely associated with cardiovascular disease. We aimed to examine the age-dependent interaction between Life's Essential 8 (LE8), the updated measurement of cardiovascular health (CVH), and CKD in the United States. The cross-sectional study involved 25,529 participants from National Health and Nutrition Examination Survey in 2007-2018. Multivariate logistic regressions were used to estimate the age-dependent interaction between LE8 and CKD, and restricted cubic spline regressions were used to analyze the dose-response relationships between LE8 and CKD among adults and all age subgroups. Overall, 2934 (9.3%), 17,278 (66.2%), and 5317 (24.5%) participants had low, moderate, and high CVH, separately. After adjusting for the potential covariates, LE8 was negatively associated with CKD [odds ratio (OR) for per 1 standard deviation (SD) increase and 95%CI, 0.71 (0.67, 0.75)], with a nonlinear dose-response relationship (P for nonlinearity = 0.001). The inversed association was stronger among participants aged 65 and older (0.65 (0.59, 0.71)) compared to youngers [20-39 years, 0.63 (0.59, 0.58), 40-64 years, 0.63 (0.59, 0.58)] (P for interaction = 0.002). CVH, as measured by the LE8 score, was negatively associated with the presence of CKD in non-linear fashions, more pronounced in participants aged 65 and older compared to younger age groups.


Thrombosis prophylaxis in the acutely ill medical patient: insights from the prophylaxis in MEDical patients with ENOXaparin (MEDENOX) trial.
The clinical benefit and cost-effectiveness of routine thromboprophylaxis in surgical patients has been well documented, but use of this approach in general medical patients remains controversial. Following is an overview of major insights from the prophylaxis in Medical patients with Enoxaparin (MEDENOX) trial, which was undertaken to evaluate the efficacy of 2 dosage regimens of the low-molecular-weight heparin enoxaparin for prevention of venous thromboembolism in acutely ill medical patients. MEDENOX was a prospective, double-blind, randomized, placebo-controlled trial, which enrolled 1,102 hospitalized patients from 68 centers in 9 countries. Patients were randomly assigned to receive 40 mg of enoxaparin, 20 mg of enoxaparin, or placebo, given subcutaneously once a day for 6-14 days. The primary outcome was venous thromboembolism (deep-vein thrombosis or pulmonary embolism) between days 1 and 14. At the 40-mg dose, the risk of venous thromboembolism was significantly reduced in patients with heart failure, as well as in patients with other medical illnesses, including respiratory failure, infectious disease, or rheumatic disorders. The incidence of venous thromboembolism was 5.5% in the group that received 40 mg of enoxaparin as compared with 14.9% in the group that received placebo (p <0.001), a benefit that was maintained at 3-month follow-up. There was no significant difference in the incidence of venous thromboembolism between the group that received 20 mg of enoxaparin (15%) and the placebo group. Adverse effects did not significantly differ between the placebo group and either enoxaparin group. This study clearly establishes an effective strategy for lowering the risk of venous thromboembolism in the acutely ill population and provides a framework for future trials in this area.


Detection and function of an intramolecular disulfide bond in the pH-responsive CadC of Escherichia coli.
In an acidic and lysine-rich environment Escherichia coli induces expression of the cadBA operon which encodes CadA, the lysine decarboxylase, and CadB, the lysine/cadaverine antiporter. cadBA expression is dependent on CadC, a membrane-integrated transcriptional activator which belongs to the ToxR-like protein family. Activation of CadC requires two stimuli, lysine and low pH. Whereas lysine is detected by an interplay between CadC and the lysine-specific transporter LysP, pH alterations are sensed by CadC directly. Crystal structural analyses revealed a close proximity between two periplasmic cysteines, Cys208 and Cys272. Substitution of Cys208 and/or Cys272 by alanine resulted in CadC derivatives that were active in response to only one stimulus, either lysine or pH 5.8. Differential in vivo thiol trapping revealed a disulfide bond between these two residues at pH 7.6, but not at pH 5.8. When Cys208 and Cys272 were replaced by aspartate and lysine, respectively, virtually wild-type behavior was restored indicating that the disulfide bond could be mimicked by a salt bridge. A disulfide bond was found in the periplasmic domain of CadC that supports an inactive state of CadC at pH 7.6. At pH 5.8 disulfide bond formation is prevented which transforms CadC into a semi-active state. These results provide new insights into the function of a pH sensor.


Neuroprotective activities of CEP-1347 in models of neuroAIDS.
When the nervous system is infected with HIV-1, it commonly results in neuroinflammation leading to overt neuronal dysfunction and subsequent cognitive and behavioral impairments. The multifaceted disease process, now referred to as HIV-1-associated neurocognitive disorders (HAND), provides a range of molecular targets for adjunctive therapies. One is CEP-1347, an inhibitor of mixed lineage kinases that elicits neuroprotective and anti-inflammatory responses in models of neurodegenerative diseases. Since HAND is associated with inflammatory encephalopathy induced by virus infection and mononuclear phagocytes (perivascular macrophages and microglia) immune activation, we investigated whether CEP-1347 could ameliorate disease in laboratory models of HAND. We now demonstrate that CEP-1347 reduces the levels of secreted proinflammatory cytokines and chemokines in HIV-1-infected human macrophages and attenuates dose-dependent neurotoxicity in rodent cortical neurons. CEP-1347-treated mice readily achieve therapeutic drug levels in peripheral blood. HIV-1 encephalitis (HIVE) mice, where human virus-infected monocyte-derived macrophages are stereotactically injected into the basal ganglia of CB17 severe combined immunodeficient mice, received daily intraperitoneal injections of CEP-1347. Here, CEP-1347 treatment of HIVE mice showed a dose-dependent reduction in microgliosis. Dendritic integrity and neuronal loss were sustained and prevented, respectively. These results demonstrate that CEP-1347 elicits anti-inflammatory and neuroprotective responses in an HIVE model of human disease and as such warrants further study as an adjunctive therapy for human disease.


The cell wall of the human pathogen Candida glabrata: differential incorporation of novel adhesin-like wall proteins.
The cell wall of the human pathogen Candida glabrata governs initial host-pathogen interactions that underlie the establishment of fungal infections. With the aim of identifying species-specific features that may directly relate to its virulence, we have investigated the cell wall of C. glabrata using a multidisciplinary approach that combines microscopy imaging, biochemical studies, bioinformatics, and tandem mass spectrometry. Electron microscopy revealed a bilayered wall structure in which the outer layer is packed with mannoproteins. Biochemical studies showed that C. glabrata walls incorporate 50% more protein than Saccharomyces cerevisiae walls and, consistent with this, have a higher mannose/glucose ratio. Evidence is presented that C. glabrata walls contain glycosylphosphatidylinositol (GPI) proteins, covalently bound to the wall 1,6-beta-glucan, as well as proteins linked through a mild-alkali-sensitive linkage to 1,3-beta-glucan. A comprehensive genome-wide in silico inspection showed that in comparison to other fungi, C. glabrata contains an exceptionally large number, 67, of genes encoding adhesin-like GPI proteins. Phylogenetically these adhesin-like proteins form different clusters, one of which is the lectin-like EPA family. Mass spectrometric analysis identified 23 cell wall proteins, including 4 novel adhesin-like proteins, Awp1/2/3/4, and Epa6, which is involved in adherence to human epithelia and biofilm formation. Importantly, the presence of adhesin-like proteins in the wall depended on the growth stage and on the genetic background used, and this was reflected in alterations in adhesion capacity and cell surface hydrophobicity. We propose that the large repertoire of adhesin(-like) genes of C. glabrata contributes to its adaptability and virulence.


In vivo transcription of the Epstein-Barr virus (EBV) BamHI-A region without associated in vivo BARF0 protein expression in multiple EBV-associated disorders.
The in vivo expression of the Epstein-Barr virus (EBV) BamHI-A rightward transcripts (BARTs) as well as the putative BART-encoded BARF0 and RK-BARF0 proteins in various EBV-associated malignancies was investigated. RT-PCRs specific for the different splice variants of the BARTs and both a nucleic acid sequence-based amplification assay and an RT-PCR specific for the BARF0 ORF were used. Abundant transcription of BARTs was found in EBV-associated Hodgkin's lymphomas, Burkitt's lymphomas (BL), T-cell non-Hodgkin's lymphomas, post-transplant lymphoproliferative disorders, AIDS-related lymphomas and gastric carcinomas. Using RNA in situ hybridization (RISH), BARTs were detected within the neoplastic cells of these malignancies. BARTs encoding RK-BARF0 were not detected. The BARTs detected were shown possibly to encode the RPMS1 and BARF0 proteins, based on their splicing. However, BARTs actually harbouring the BARF0 ORF were detected only in specimens containing a relatively large number of EBV-positive cells. New monoclonal antibodies against the BARF0 protein were generated that efficiently recognized prokaryotic and eukaryotic recombinant BARF0. However, the BARF0 protein was not detected in clinical samples, nor in EBV-positive cell lines, even though these were positive for BARTs by RISH and/or BARF0 RNA in vitro analysis. Using immunoblot analysis, no antibodies against baculovirus-expressed BARF0 protein were detected in the sera of nasopharyngeal carcinoma patients, BL patients and Hodgkin's disease patients, patients with chronic EBV infection, infectious mononucleosis patients or EBV-positive healthy donors. Thus, BARTs containing the BARF0 ORF are expressed in vivo but the BARF0 protein cannot be detected and may be expressed only marginally. It is concluded that the BARF0 protein is unlikely to play a role in vivo in EBV-positive malignancies.


n-3 essential fatty acid and vitamin D supplementation improve skeletal health in laying hens.
Keel bone fractures and osteoporosis are prevalent and damaging skeletal issues in the laying hen industry. There is a large interest in improving bone quality parameters to reduce or eliminate these conditions, thus improving bird welfare. Both essential fatty acids (EFA) and vitamin D can play a role in bone metabolism. The hypothesis of this study was that birds supplemented with lower n-6:n-3 EFA ratio or vitamin D would have improved bone properties compared to a control diet. A total of 3,520 Lohmann Brown-Lite pullets were used in this study. Pullets were housed on the floor from 0 to 17 wk of age and then moved to an aviary (17-52 wk of age). Starting at 12 wk of age, birds were split into diet treatments-control, flax, fish, or vitamin D diets with n-6:n-3 ratios of 6.750, 0.534, 0.534, and 6.750, respectively. Diets were formulated to be isonitrogenous and isocaloric. Basal vitamin D3 levels were formulated to be 2,760 IU/kg across all diets; for the vitamin D diet, the vitamin D3 level was increased to 5,520 IU/kg. Hens on fish and vitamin D diets had greater bone density, keel bone volume, digital bone mineral content, and keel condition compared to flax and control hens. Additionally, birds fed the vitamin D diet had the heaviest body weights compared to birds fed fish or control diets. Birds fed the flax and vitamin D diets had improved feather coverage across multiple body regions. Feeding an n-3 EFA- or vitamin D-enriched diet decreased mortality by 1.6 to 3.3% compared to the control. The fish and vitamin D diets generated mixed production performance. Compared to the other treatments, the vitamin D diet generated higher case weights but lower hen day percentage throughout the study. When compared to the other treatments, the fish diet had the lowest case weights but had a greater hen day percentage after 36 wk of age. Results indicate that a fish-based EFA and vitamin D supplementation show promise in improving skeletal health but require further investigation.


Clinical tolerance in allogeneic hematopoietic stem cell transplantation.
Allogeneic hematopoietic stem cell transplantation (HSCT) has been a curative therapeutic option for a wide range of immune hematologic malignant and non-malignant disorders including genetic diseases and inborn errors. Once in the host, allogeneic transplanted cells have not only to ensure myeloid repopulation and immunological reconstitution but also to acquire tolerance to host human leukocyte antigens via central or peripheral mechanisms. Peripheral tolerance after allogeneic HSCT depends on several regulatory mechanisms aimed at blocking alloimmune reactivity while preserving immune responses to pathogens and tumor antigens. Patients transplanted with HSCT represent an ideal model system in humans to identify and characterize the key cellular and molecular players underlying these mechanisms. The knowledge gained from these studies has allowed the development of novel therapeutic strategies aimed at inducing long-term peripheral tolerance, which can be applicable not only in allogeneic HSCT but also in autoimmune diseases and solid-organ transplantation. In the present review, we describe Type 1 regulatory T cells, initially discovered and characterized in chimeric patients transplanted with human leukocyte antigen-mismatched HSCT, and how their presence correlates to tolerance induction and maintenance. Furthermore, we summarize different cell therapy approaches with regulatory T cells, designed to facilitate tolerance induction, minimizing pharmaceutical interventions.


Radiology at war.
It can be assumed that the high percentage of wounded sent immediately from the triage area to the department of radiology was a reflection of the necessity for X-rays. The radiological examination was essential in reaching the correct diagnosis, and the radiologist was the central consultant, as by giving the diagnostic information and performing therapeutic angiographic intervention, he actively participated in the treatment of the patient. The radiologist and the surgeon together reached the final decision as to which injury prevailed and in which sequence treatment had to be planned. Hence, the department of radiology became the vital link in flow of patients from the sorting area to the operating theatre.


Clostridium perfringens enterotoxin in antibiotic-associated diarrhea.
Clostridium perfringens type A is associated with 5-20% cases of antibiotic-associated diarrhea (AAD) even though Clostridium difficile is implicated in the most severe cases. Fecal specimens from one hundred hospitalized patients, who developed diarrhea regardless of antibiotic intake and who were negative for C. difficile toxin assay, were investigated for C. perfringens enterotoxin (CPE). Simultaneously, cultures were set up for other possible aetiological factors. Ten healthy controls were also similarly investigated. CPE was positive in 2/100 (2%) of the patients and the samples were also positive for the organism in culture. Other organisms isolated were non-toxigenic C. difficile (4%), staphylococci (6%), Candida (18%) and Klebsiella pneumoniae (1%). Stool samples from healthy controls grew mixed growth of no significance and CPE was negative in all of them. Detection of CPE is not part of routine laboratory investigation due to resource implication. Criteria for initiating investigations have to be therefore established by understanding the true burden of C. perfringens-associated AAD by further research.


Epstein-Barr Virus Genomes Reveal Population Structure and Type 1 Association with Endemic Burkitt Lymphoma.
Endemic Burkitt lymphoma (eBL), the most prevalent pediatric cancer in sub-Saharan Africa, is distinguished by its inclusion of Epstein-Barr virus (EBV). In order to better understand the impact of EBV variation in eBL tumorigenesis, we improved viral DNA enrichment methods and generated a total of 98 new EBV genomes from both eBL cases (n = 58) and healthy controls (n = 40) residing in the same geographic region in Kenya. Using our unbiased methods, we found that EBV type 1 was significantly more prevalent in eBL patients (74.5%) than in healthy children (47.5%) (odds ratio = 3.24, 95% confidence interval = 1.36 to 7.71, P = 0.007), as opposed to similar proportions in both groups. Controlling for EBV type, we also performed a genome-wide association study identifying six nonsynonymous variants in the genes EBNA1, EBNA2, BcLF1, and BARF1 that were enriched in eBL patients. In addition, viruses isolated from plasma of eBL patients were identical to their tumor counterparts consistent with circulating viral DNA originating from the tumor. We also detected three intertypic recombinants carrying type 1 EBNA2 and type 2 EBNA3 regions, as well as one novel genome with a 20-kb deletion, resulting in the loss of multiple lytic and virion genes. Comparing EBV types, viral genes displayed differential variation rates as type 1 appeared to be more divergent, while type 2 demonstrated novel substructures. Overall, our findings highlight the complexities of the EBV population structure and provide new insight into viral variation, potentially deepening our understanding of eBL oncogenesis.IMPORTANCE Improved viral enrichment methods conclusively demonstrate EBV type 1 to be more prevalent in eBL patients than in geographically matched healthy controls, which previously underrepresented the prevalence of EBV type 2. Genome-wide association analysis between cases and controls identifies six eBL-associated nonsynonymous variants in EBNA1, EBNA2, BcLF1, and BARF1 genes. Analysis of population structure reveals that EBV type 2 exists as two genomic subgroups and was more commonly found in female than in male eBL patients.


Diagnosis of tears in rotator-cuff-injuries.
Pathology of the rotator cuff is the cause of most common problems at the shoulder joint. Acute injuries are not as frequent as chronic cuff disease, but often they aggravate inflammatory or degenerative tendon alterations, even if they are of minor severity. Traumatic rotator cuff tears predominantly affect the supraspinatus tendon or the rotator interval. The subscapularis tendon is involved in anterior dislocations of the glenohumeral joint or in direct trauma. Plain film radiography still remains the base of all further imaging studies. If only full-thickness tears must be ruled out, double-contrast arthrography and ultrasound are acceptable imaging modalities. However, the former has a drawback in being invasive and does not detect partial tears at the bursal site of the cuff or rotator cuff tendinopathy, whereas the latter heavily depends on the experience of the radiologist and is restricted to the rotator cuff. Nowadays the most comprehensive imaging method is magnetic resonance (MR) imaging. MR imaging enables the detection or exclusion of complete rotator cuff tears with a reasonable accuracy and is also suitable to diagnose further pathologies of the shoulder joint. MR arthrography is valuable in the detection of subtle anatomic details and further improves the differentiation of rotator cuff diseases. Although in comparison MR imaging is still the most expensive imaging method, its high negative predictive value for the diagnosis of complete rotator cuff tears and its reliability evaluating different shoulder joint pathologies make it the preferred imaging modality.


Mutation of FA2H underlies a complicated form of hereditary spastic paraplegia (SPG35).
Hereditary spastic paraplegia (HSP) describes a heterogeneous group of inherited neurodegenerative disorders in which the cardinal pathological feature is upper motor neurone degeneration leading to progressive spasticity and weakness of the lower limbs. Using samples from a large Omani family we recently mapped a gene for a novel autosomal recessive form of HSP (SPG35) in which the spastic paraplegia was associated with intellectual disability and seizures. Magnetic resonance imaging of the brain of SPG35 patients showed white matter abnormalities suggestive of a leukodystrophy. Here we report homozygous mutations in the fatty acid 2-hydroxylase gene (FA2H) in the original family used to define the SPG35 locus (p.Arg235Cys) as well as in a previously unreported Pakistani family with a similar phenotype (p.Arg53_Ile58del). Measurement of enzyme activity in vitro revealed significantly reduced enzymatic function of FA2H associated with these mutations. These results demonstrate that mutations in FA2H are associated with SPG35, and that abnormal hydroxylation of myelin galactocerebroside lipid components can lead to a severe progressive phenotype, with a clinical presentation of complicated HSP and radiological features of leukodystrophy. (c) 2010 Wiley-Liss, Inc.


Promoting vibrations in human purine nucleoside phosphorylase. A molecular dynamics and hybrid quantum mechanical/molecular mechanical study.
Crystallographic studies of human purine nucleoside phosphorylase (hPNP) with several transition-state (TS) analogues in the immucillin family showed an unusual geometric arrangement of the atoms O-5', O-4', and O(P), the nucleophilic phosphate oxygen, lying in a close three-oxygen stack. These observations were corroborated by extensive experimental kinetic isotope effect analysis. We propose that protein-facilitated dynamic modes in hPNP cause this stack, centered on the ribosyl O-4' oxygen, to squeeze together and push electrons toward the purine ring, stabilizing the oxacarbenium character of the TS. As the N-ribosidic bond is cleaved during the reaction, the pK(a) values of N-7 and O-6 increase by the electron density expelled by the oxygen-stack compression toward the purine ring. Increased electron density in the purine ring improves electrostatic interactions with nearby residues and facilitates the abstraction of a proton from a solvent proton or an unidentified general acid, making the purine a better leaving group, and accelerating catalysis. Classical and mixed quantum/classical molecular dynamics (MD) simulations of the Michaelis complex of hPNP with the substrates guanosine and phosphate were performed to assess the existence of protein-promoting vibrations (PPVs). Analogous simulations were performed for the substrates in aqueous solution. In the catalytic site, the O-5', O-4', and O(P) oxygens vibrate at frequencies of ca. 125 and 465 cm(-1), as opposed to 285 cm(-1) in the absence of hPNP. The hybrid quantum mechanical/molecular mechanical method was used to assess whether this enzymatic vibration pushing the oxygens together is coupled to the reaction coordinate, and thus has a direct positive impact on catalysis. The potential energy surface for the phosphorolysis reaction for several snapshots taken from the classical MD simulation showed substantial differences in oxygen compression. Our calculations showed the existence of PPVs coupled to the reaction coordinate, which effect electronic alterations in the active site by pushing the three oxygen centers together in proximity, and accelerate substrate turnover in the phosphorolysis reaction catalyzed by hPNP.


Phosphopyruvate carboxylase induction by L-tryptophan. Effects on synthesis and degradation of the enzyme.
1. The administration of l-tryptophan to fed rats produces a twofold increase in hepatic phosphopyruvate carboxylase activity that represents a comparable increase in enzyme protein. With specific antibody against the enzyme we have shown that the increase in phosphopyruvate carboxylase is partially mediated via an actinomycin D-sensitive increase in enzyme synthesis. 2. In starved animals tryptophan increases the enzyme activity without any change in the relative rate of phosphopyruvate carboxylase synthesis. In this condition degradation of the enzyme is retarded by tryptophan by a mechanism that is not prevented by cycloheximide.


Plasmonic monitoring of catalytic hydrogen generation by a single nanoparticle probe.
Plasmonic nanostructures such as gold nanoparticles are very useful for monitoring chemical reactions because their optical properties are highly dependent upon the environment surrounding the particle surface. Here, we designed the catalytic structure composed of platinized cadmium sulfide with gold domains as a sensitive probe, and we monitored the photocatalytic decomposition of lactic acid to generate hydrogen gas in situ by single-particle dark-field spectroscopy. The plasmon band shift of the gold probe throughout the reaction exhibits significant particle-to-particle variation, and by simulating the reaction kinetics, the rate constant and structural information (including the diffusion coefficient through the shell and the relative arrangement of the active sites) can be estimated for individual catalyst particles. This approach is versatile for the monitoring of various heterogeneous reactions with distinct components at a single-particle level.


Lymphoma-associated skin cancer: incidence, natural history, and clinical management.
The link between immunosuppression and skin cancer has been well described. The two most common situations involving immunosuppression-associated skin cancer are solid organ transplantation and non-Hodgkin lymphoma (NHL), including chronic lymphocytic leukemia (CLL). Patients with lymphoma are more likely to have development of a secondary malignancy, with skin cancer being the most common. The most common types of skin cancer in patients with NHL/CLL include melanoma, squamous cell carcinoma, basal cell carcinoma, and Merkel cell carcinoma. Many skin cancers demonstrate increased aggressiveness in patients with NHL/CLL and are associated with higher recurrence rates, increased regional metastasis, and death secondary to skin cancer metastases. This review delineates the current research regarding the relationship between NHL/CLL and cutaneous malignancy. Immunosuppressed patients with skin cancer should be treated promptly and aggressively to decrease recurrence and metastases. Regular skin self-examinations, dermatologic examinations, sun-protective habits, and education may prove beneficial in this high-risk patient population.


[Glomerulonephritis secondary to ventriculo-atrial shunts].
The authors report the case of a 45 year old man who had undergone a ventriculo-atrial CSF shunt procedure 5 years previously for normal pressure hydrocephalus and who had several unexplained episodes of infection over a 12 months period and has now developed a mixed nephrotic syndrome associated with a septicaemia. Corynebacterium commensale and Staphylococcus epidermis were isolated from the valve culture. Ablation of the valve resulted in clinical cure with minimal functional renal sequellae. The initial renal biopsy showed type I proliferative glomerulonephritis with subendothelial deposits of complement and immunoglobulins, which did not completely regress after 3 months' evolution. The serum complement fractions suggested activation of the alternate pathway and the possible pathogenic role of circulating immune complexes.


[Cytological diagnosis of prostatic cancer by transrectal aspiration biopsy (author's transl)].
From the cytologic study of fifty transrectal needle aspirations of the prostate the authors try to establish the principles leading to the diagnosis of prostatic carcinoma. Correlation with clinical diagnosis shows that in positive cases, cytologic diagnosis is accurate. In the case of negative cytologic responses this negative response may be imputable: to the paucicellularity of the sample, unavoidable blind needle aspiration, an early necrosis of the cellular material, a false interpretation of minimal cellular and nuclear abnormalities. This method proves valuable for the clinician. In two cases out of three the cytological diagnosis confirms the clinical diagnosis.


Efficacy of cardiac rehabilitation services. With emphasis on patients after myocardial infarction.
During the 1970s, emphasis increased in clinical practice on early ambulation and exercise-based rehabilitation after myocardial infarction and other cardiac illnesses or procedures. This shift was based on the belief that exercise and improved conditioning would improve prognosis. We examine the evidence supporting this assertion. Most of the reports on cardiac rehabilitation are about patients who have coronary artery disease and a history of myocardial infarction. The review, therefore, is focused primarily on the patient who has had a myocardial infarction. Effects of cardiac rehabilitation, emphasizing exercise treatment and conditioning, are reviewed with regard to patient outcomes, including changes in functional (work) capacity, psychosocial functioning and health-related knowledge, risk factor modification, morbidity and mortality, and cardiac function. The safety of cardiac exercise programs is reviewed, and the use of telemetry monitoring is considered. We also discuss the role of cardiac rehabilitation in categories of patients other than those with myocardial infarction and the application of newer approaches to rehabilitation such as programs based in the patient's home.


Metabolic acidosis in the critically ill: part 1. Classification and pathophysiology.
Metabolic acidaemia (pH < 7.35 not primarily related to hypoventilation) is common amongst the critically ill and it is essential that clinicians caring for such patients have an understanding of the common causes. The exclusive elimination routes of volatile (carbon dioxide), organic (lactic and ketone) and inorganic (phosphate and sulphate) acids mean compensation for a defect in any one is limited and requires separate provision during critical illness. We discuss the models available to diagnose metabolic acidosis including CO2/HCO3(-) and physical chemistry-derived (Stewart or Fencl-Stewart) approaches, but we propose that the base excess and anion gap, corrected for hypoalbuminaemia and iatrogenic hyperchloraemia, remain most appropriate for clinical usage. Finally we provide some tips for interpreting respiratory responses to metabolic acidosis and how to reach a working diagnosis, the consequences of which are considered in Part 2 of this review.


The Onset of Urinary Incontinence in Different Subgroups and its Relation to Menopausal Status: A Hospital-based Study.
Numerous risk factors have been reported to influence the development of urinary incontinence (UI). In this study, we took a closer look on the different forms of UI and tried to identify differences in regard to potential risk factors. Of special interest was the onset of UI symptoms and its relation to menopausal status. This was a hospital-based analysis of patients who presented with urinary incontinence in the outpatient ward of a tertiary hospital. The diagnosis of urinary incontinence was based on the subjective complaints of patients. Data concerning menopausal status, hormone replacement therapy, prior hysterectomy were assessed. The mean age was 53.8 years in the SUI group, 62.7 years in the MUI group and 66.1 years in the UUI group, respectively (p<0.001). The proportion of patients with UUI was higher in the postmenopausal group, whereas the proportion of SUI was higher in the premenopausal group (p<0.001). The mean age in which complaints occurred was significantly lower in the SUI group (45.4 years) compared to the MUI (51.0 years) and UUI groups (54.7 years) (p<0.001). There was no correlation between menopausal status and onset of urinary incontinence (p=0.143). Additional anamnestic information help further characterize the different types of urinary incontinence that can lead to an optimization of treatment options. Younger age and premenopausal status were accompanied by milder forms of UI while menopausal status itself had no influence on the onset of UI symptoms indicating that age-related changes may lead to different types of incontinence.


How useful is quantitative risk assessment?
This article discusses the use of quantitative risk assessment (QRA) in decision making regarding the safety of complex technological systems. The insights gained by QRA are compared with those from traditional safety methods and it is argued that the two approaches complement each other. It is argued that peer review is an essential part of the QRA process. The importance of risk-informed rather than risk-based decision making is emphasized. Engineering insights derived from QRAs are always used in combination with traditional safety requirements and it is in this context that they should be reviewed and critiqued. Examples from applications in nuclear power, space systems, and an incinerator of chemical agents are given to demonstrate the practical benefits of QRA. Finally, several common criticisms raised against QRA are addressed.


The effects of swimming training on arterial function, muscular strength, and cardiorespiratory capacity in postmenopausal women with stage 2 hypertension.
Aging is associated with progressive decreases in arterial health as well as muscular strength and cardiorespiratory capacity. It is crucial to prevent or reduce the negative effects of aging on vasculature, muscular, and cardiorespiratory function by implementing appropriate lifestyle interventions, such as exercise training. We examined the effects of a swimming (SWM) regimen on arterial stiffness (pulse wave velocity [PWV]), blood pressure (BP), wave reflection (augmentation index [AIx]), muscle strength, and aerobic capacity in sedentary, postmenopausal women with stage 2 hypertension. One hundred participants (∼74 years of age) were recruited from multiple general practice centers. Using a parallel experimental design, participants were randomly assigned to either SWM (n = 52) or nonexercising control group (n = 48) for 20 weeks. Participants in the SWM group trained 3 to 4 days/week. Participants' carotid to radial PWV, BP, AIx, muscular strength, and cardiorespiratory capacity were measured at baseline and after 20 weeks of their assigned intervention. There was a significant group × time interaction (P < 0.05) for carotid to radial PWV (Δ-1.2 m/s), AIx (Δ-4%), and systolic (Δ-9 mm Hg) and diastolic BP (Δ - 9 mm Hg), which significantly decreased (P < 0.05); and strength (Δ3 kg) and cardiorespiratory capacity (Δ4 mL/kg/min), which significantly increased (P < 0.05) following SWM compared to no changes in control. SWM led to reductions in arterial stiffness, wave reflection, and BP while increasing strength and aerobic capacity in postmenopausal women with stage 2 hypertension. SWM may be an effective intervention in the prevention and treatment of age-related vascular complications and declines in muscle strength and cardiorespiratory capacity.


[Cyclooxygenase(COX)-2-inhibition in the prevention and treatment of colorectal carcinoma].
Epidemiological studies have found that long-term use of non-steroidal anti-inflammatory drugs (NSAIDs) is associated with a lower risk of colorectal cancer (CRC). Cyclooxygenase (COX)-2 expression is present in colorectal cancer and overexpression is associated with metastases and poorer prognosis in multivariate analysis. NSAID treatment results in a reduction of the incidence of colorectal adenoma in patients with familial adenomatous polyposis, in patients with a history of colorectal adenomas and in patients with a history of CRC. Pre-clinical research shows that COX-2 expression is associated with cell proliferation, angiogenesis, apoptosis inhibition and local immune-down modulation. An anticarcinogenic effect has been shown specifically in selective COX-2 inhibitors in animal models. Selective COX-2 inhibitors have fewer adverse effects than the non-selective NSAIDs and are promising chemopreventative and chemotherapeutical agents. The effects of selective COX-2 inhibition in the prevention of and treatment for colorectal carcinoma will be investigated in clinical randomized multicentre trials.


Surgical treatment for metastatic malignancies. Pulmonary metastasis: indications and outcomes.
Surgical resection is an important modality in the treatment of pulmonary metastases from various solid tumors. The criteria for pulmonary metastasectomy are as follows: (1) the patient must be a good risk for surgical intervention; (2) the primary malignancy is controlled; (3) there is no other, extrapulmonary, metastasis; and (4) the pulmonary lesions are thought to be completely resectable. The appropriate selection of candidates according to these criteria leads to an overall 5-year survival after pulmonary metastasectomy of about 30%-40%. However, most of the reported results are retrospective analyses, and the significance of pulmonary metastasectomy seems to vary according to the primary malignancy. To clarify the significance of surgical treatment for pulmonary metastases, we need further analysis of various prognostic factors, with special reference to each primary malignancy, as well as a multiinstitutional study, and randomized prospective studies, if possible.


Cutaneous and Mucosal Manifestations Associated with Celiac Disease.
Celiac disease (CD) is an immune-mediated, gluten-induced enteropathy that affects predisposed individuals of all ages. Many patients with CD do not report gastrointestinal symptoms making it difficult to reach an early diagnosis. On the other hand, CD is related to a wide spectrum of extra-intestinal manifestations, with dermatitis herpetiformis (DH) being the best characterized. These associated conditions may be the clue to reaching the diagnosis of CD. Over the last few years, there have been multiple reports of the association between CD and several cutaneous manifestations that may improve with a gluten-free diet (GFD). The presence of some of these skin diseases, even in the absence of gastrointestinal symptoms, should give rise to an appropriate screening method for CD. The aim of this paper is to describe the different cutaneous manifestations that have been associated with CD and the possible mechanisms involved.


Spatial differences in East scotia ridge hydrothermal vent food webs: influences of chemistry, microbiology and predation on trophodynamics.
The hydrothermal vents on the East Scotia Ridge are the first to be explored in the Antarctic and are dominated by large peltospiroid gastropods, stalked barnacles (Vulcanolepas sp.) and anomuran crabs (Kiwa sp.) but their food webs are unknown. Vent fluid and macroconsumer samples were collected at three vent sites (E2, E9N and E9S) at distances of tens of metres to hundreds of kilometres apart with contrasting vent fluid chemistries to describe trophic interactions and identify potential carbon fixation pathways using stable isotopes. δ(13)C of dissolved inorganic carbon from vent fluids ranged from -4.6‰ to 0.8‰ at E2 and from -4.4‰ to 1.5‰ at E9. The lowest macroconsumer δ(13)C was observed in peltospiroid gastropods (-30.0‰ to -31.1‰) and indicated carbon fixation via the Calvin-Benson-Bassham (CBB) cycle by endosymbiotic gamma-Proteobacteria. Highest δ(13)C occurred in Kiwa sp. (-19.0‰ to -10.5‰), similar to that of the epibionts sampled from their ventral setae. Kiwa sp. δ(13)C differed among sites, which were attributed to spatial differences in the epibiont community and the relative contribution of carbon fixed via the reductive tricarboxylic acid (rTCA) and CBB cycles assimilated by Kiwa sp. Site differences in carbon fixation pathways were traced into higher trophic levels e.g. a stichasterid asteroid that predates on Kiwa sp. Sponges and anemones at the periphery of E2 assimilated a proportion of epipelagic photosynthetic primary production but this was not observed at E9N. Differences in the δ(13)C and δ(34)S values of vent macroconsumers between E2 and E9 sites suggest the relative contributions of photosynthetic and chemoautotrophic carbon fixation (rTCA v CBB) entering the hydrothermal vent food webs vary between the sites.


Novel 2D and 3D Assays to Determine the Activity of Anti-Leishmanial Drugs.
The discovery of novel anti-leishmanial compounds remains essential as current treatments have known limitations and there are insufficient novel compounds in development. We have investigated three complex and physiologically relevant in vitro assays, including: (i) a media perfusion based cell culture model, (ii) two 3D cell culture models, and (iii) iPSC derived macrophages in place of primary macrophages or cell lines, to determine whether they offer improved approaches to anti-leishmanial drug discovery and development. Using a Leishmania major amastigote-macrophage assay the activities of standard drugs were investigated to show the effect of changing parameters in these assays. We determined that drug activity was reduced by media perfusion (EC50 values for amphotericin B shifted from 54 (51-57) nM in the static system to 70 (61-75) nM under media perfusion; EC50 values for miltefosine shifted from 12 (11-15) µM in the static system to 30 (26-34) µM under media perfusion) (mean and 95% confidence intervals), with corresponding reduced drug accumulation by macrophages. In the 3D cell culture model there was a significant difference in the EC50 values of amphotericin B but not miltefosine (EC50 values for amphotericin B were 34.9 (31.4-38.6) nM in the 2D and 52.3 (46.6-58.7) nM in 3D; EC50 values for miltefosine were 5.0 (4.9-5.2) µM in 2D and 5.9 (5.5-6.2) µM in 3D (mean and 95% confidence intervals). Finally, in experiments using iPSC derived macrophages infected with Leishmania, reported here for the first time, we observed a higher level of intracellular infection in iPSC derived macrophages compared to the other macrophage types for four different species of Leishmania studied. For L. major with an initial infection ratio of 0.5 parasites per host cell the percentage infection level of the macrophages after 72 h was 11.3% ± 1.5%, 46.0% ± 1.4%, 66.4% ± 3.5% and 75.1% ± 2.4% (average ± SD) for the four cells types, THP1 a human monocytic cell line, mouse bone marrow macrophages (MBMMs), human bone marrow macrophages (HBMMs) and iPSC derived macrophages respectively. Despite the higher infection levels, drug activity in iPSC derived macrophages was similar to that in other macrophage types, for example, amphotericin B EC50 values were 35.9 (33.4-38.5), 33.5 (31.5-36.5), 33.6 (30.5-not calculated (NC)) and 46.4 (45.8-47.2) nM in iPSC, MBMMs, HBMMs and THP1 cells respectively (mean and 95% confidence intervals). We conclude that increasing the complexity of cellular assays does impact upon anti-leishmanial drug activities but not sufficiently to replace the current model used in HTS/HCS assays in drug discovery programmes. The impact of media perfusion on drug activities and the use of iPSC macrophages do, however, deserve further investigation.


Detection of sonographic B-lines in patients with normal lung or radiographic alveolar consolidation.
Diffuse comet-tail B-line artifacts in lung ultrasound are a sign of alveolar-interstitial syndrome, but isolated transthoracic scans positive for B-lines (multiple B lines or B+) could be detected in other conditions. The aim was to assess the prevalence and distribution of this sonographic sign in patients with normal lung or isolated alveolar consolidation in chest radiography. Two hundred seventeen patients consecutively admitted to this emergency medicine unit with any diagnosis and without radiographic or clinical evidence of diffuse interstitial syndrome were analyzed. Each patient underwent chest radiography and lung sonographic examination with four anterolateral chest scans per side. Of the 1736 sonographic scans performed, 13.2% were positive for the B+ pattern. Positive scans significantly corresponded to laterobasal areas or radiographic opacities due to lung alveolar consolidations (p<0.005). Twenty percent of the laterobasal scans of 145 patients with radiologically normal lung were positive. The negative predictive value of B+ was 83.9% (95% confidence interval: 78.2-89.7%), with a specificity of 90.3% (95%CI: 85.5-95.1%) for predicting any localized radiographic pulmonary opacity. B+ scans can be detected in the chest areas surrounding an isolated alveolar consolidation and in the laterobasal scans of a radiographic normal lung. These features should always be considered when lung ultrasound is performed to rule out the alveolar-interstitial syndrome in an emergency setting. Moreover, B+ patterns have a satisfactory negative predictive value for radiographic lung opacities, which could have added diagnostic value in the ED evaluation of dyspneic patients.