Effects of Analgesics on Tumor Growth in Mouse Models of Prostate Cancer Bone Metastasis.
Murine models of tumor development often require invasive procedures for tumor implantation, potentially causing pain or distress. However, analgesics are often withheld during implantation because of concerns that they may adversely affect tumor development. Previous studies examining the effects of analgesics on the development and metastasis of various tumor lines show that the effect of analgesics depends on the tumor line and analgesic used. A blanket statement that analgesics affect the general growth of tumors is not adequate scientific justification for withholding pain relief, and pilot studies or references are recommended for each specific tumor cell line and treatment combination. In this study, we evaluated the effects of 2 commonly used analgesics on tumor growth in 2 models of prostate cancer (PCa) bone metastasis. We hypothesized that a one-time injection of analgesics at the time of intratibial injection of tumor cells would not significantly impact tumor growth. Either C57BL/6 or SCID mice were injected subcutaneously with an analgesic (carprofen [5 mg/kg], or buprenorphine [0.1 mg/kg]) or vehicle (0.1 mL of saline) at the time of intratibial injection with a PCa cell line (RM1 or PC3, n = 10 to 11 per group). Tumor growth (measured by determination of tumor burden and the extent of bone involvement) and welfare (measured by nociception, locomotion, and weight) were monitored for 2 to 4 wk. Neither carprofen or buprenorphine administration consistently affected tumor growth or indices of animal welfare as compared with the saline control for either cell line. This study adds to the growing body of literature demonstrating that analgesia can be compatible with scientific objectives, and that a decision to withhold analgesics must be scientifically justified and evaluated on a model-specific basis.


Feasibility of quantitative ultrasound measurement of the heel bone in people with intellectual disabilities.
Low bone mineral density (BMD) and fractures are common in people with intellectual disabilities (ID). Reduced mobility in case of motor impairment and the use of anti-epileptic drugs contribute to the development of low BMD. Quantitative ultrasound (QUS) measurement of the heel bone is a non-invasive and radiation-free method for measuring bone status that can be used outside the hospital. QUS might be used for screening purposes to identify people with intellectual disability with poor bone status, who are in need of supplementary examination and treatment. To investigate feasibility of QUS in this group, QUS of the heel bone was performed on-site in 151 people with ID living in residential care. Measurements were successfully performed in at least one foot in 94.7%, were interpretable (resulting in a stiffness index) in 91.6%, and induced barely or no stress in 90.4% of the study population. Measurements generally took less than 10 min. In 93 persons bone status of both feet had been measured. The "mean percentage of the absolute difference" between outcomes of both feet was 15.5% (±15.3% SD, range 0-76.5%). Ultrasound measurement of the heel bone is a feasible and non-stressful method for measuring bone status in people with ID. Since the mean difference between outcomes of the left and right foot were large, measurement of both feet is recommended to prevent inaccurate interpretation.


Portal hypertension associated with hepatocellular carcinoma with cirrhosis.
Portal hypertension associated with hepatocellular carcinoma in the absence of cirrhosis of the liver has not been clearly recognized in the past. While the incidence of this association is unknown, its occurrence does not appear to be rare. This case report describes a 49-year old male with hepatocellular carcinoma, ascites and measured portal hypertension but no cirrhosis of the liver. The portal hypertension was secondary to microscopic invasion of central veins and small portal veins. Hepatocellular carcinoma in the absence of cirrhosis should be added to the differential diagnosis of portal hypertension.


Forgetting and recovering the unforgettable.
Two experiments demonstrated striking, reversible forgetting effects that occurred even for a list of expletives. The experiments used a procedure based on the classic memory mechanisms of interference and retrieval cuing. Interference reduced recall dramatically, although appropriate cues triggered complete recovery. Distinctive, emotionally charged materials were quite susceptible to the forgetting and recovery effects. Thus, powerful forgetting effects can be obtained when participants have no intentions to forget and the materials involved are distinctive, emotional materials with sexual and violent content. This forgetting is reversible with appropriate cues. The false-memory debate can and must be informed by experimental investigations not only of false memories, but also of blocked and recovered memories.


Inequalities in disability-free life expectancy by social class and area type: England, 2001-03.
Disability-free life expectancy (DFLE) is an important indicator which combines longevity with functional health status. This article examines inequalities in DFLE by socio-economic position in England, and between Local Authorities (LAs) in the deprived 'Spearhead group' and other LAs. Census and vital event data available from the ONS Longitudinal Study were used to calculate estimates of DFLE based on limiting long-term illness or disability status for each Registrar General's Social Class (RGSC) in 2001-03, in England as a whole and within the 'Spearhead group' and non-Spearhead LAs. A predominantly linear relationship was present with DFLE increasing with rising social class. The differences observed in DFLE at birth and at age 65 between people assigned to the professional and unskilled manual social classes were statistically significant and substantial, demonstrating a clear social inequality in the amount of life, the functional health status during those years lived, their absolute number, and thus the relative proportion of life spent free from a limiting long-term illness or disability.Among professional men, no statistically significant differences in DFLE at birth were found between the 'Spearhead group' and the non-Spearhead LAs. However, men assigned to the other social classes in the 'Spearhead group' were disadvantaged in terms of lower DFLE than their counterparts in the non-Spearhead LAs. Manual women living in non-spearhead LAs had comparable DFLE to women assigned to the managerial and technical and skilled non-manual social classes living in the 'Spearhead group'.The magnitude of inequality in DFLE at birth between professionals and the unskilled manual class in the proportion of life spent free of a limiting long-term illness or disability also varied between the 'Spearhead group' and non-Spearhead LAs: among men, a difference of 7.7 per cent was present in non-Spearhead LAs, compared with 13.1 per cent in the 'Spearhead group'; among women, the equivalent differences were 5.2 per cent and 9.4 per cent. These results confirm that the assumptions made with regard to sex differences in DFLE need to be set in the context of socio-economic position, and assumptions made regarding sex and social differences need to set in the context of geography. The scale of the inequalities shown is important for policy responses and understanding of differences in service needs.


Porcine cardiac valvular endothelial cells in culture. A relative deficiency of fibronectin synthesis in vitro.
We established culture lines of porcine endothelial cells derived from both the aortic valve and the ascending aorta. These cells had the typical morphology of vascular endothelial cells in culture and contained angiotensin-converting enzyme activity. The cells derived from the aorta grew well in standard tissue culture dishes; the valvular endothelial cells, however, could not be passed serially in vitro unless cultivated in dishes precoated with fibronectin. Moreover, under the conditions used, the valvular cells showed substantial changes in their morphology when deprived of fibronectin coating for several passages but reestablished typical vascular endothelial morphology when subcultured to fibronectin-coated dishes. The aortic cells did not show this behavior. Endogenous labeling of proteins synthesized in vitro by the two cell types with 35S-methionine revealed that a high molecular weight protein produced in substantial quantities by the aortic cells was synthesized only in trace amounts by the valvular cells. This protein could be isolated by gelatin-agarose chromatography and showed an apparent molecular weight under reduced conditions of 235,000 to 245,000 on sodium dodecyl sulfate gel electrophoresis. Synthesis of gelatin-binding proteins, an indirect estimation of fibronectin production, as well as direct measurement of porcine fibronectin present in the culture supernatants using an immunoradiometric assay, showed that aortic endothelial cells synthesized and released 10-fold more fibronectin in vitro than did valvular endothelial cells. We suggest that this relative deficiency of in vitro fibronectin synthesis by valvular cells, if paralleled in vivo, may have important implications for the pathogenesis of lesions unique to the cardiac valve.


Intracellular temperature sensing by a ratiometric fluorescent polymer thermometer.
Intracellular temperature imaging could help to understand diverse biological reactions and functions in living cells. Herein, we report a nanothermometer based on ratiometric fluorescent polymers (RFPs) synthesized by the living/controlled reversible addition-fragmentation transfer polymerization method for intracellular temperature sensing. The thermometer was composed of a thermo sensitive polymer, a polarity sensitive fluorescent dye and a thermo insensitive fluorescent dye. With the increasing temperature, the fluorescent intensity of RFPs increased because of the fluorescent intensity enhancement of the polarity sensitive fluorescent dye induced by the self assembling of the thermo sensitive polymer. The prepared RFPs exhibited a fluorescence "turn-on" response at higher temperature. We further investigated the simultaneous temperature sensing and the ratiometric imaging of temperature variations associated with biological processes in living cells using this novel ratiometric probe. The polymer based ratiometric fluorescent thermometer can be used to precisely measure the temperature in living cells, and shows great potential in spatio-temporal temperature sensing down to the nanoscale within biological systems.


Composition of pigmented centers of cholesterol gallstones.
Most cholesterol gallstones have visually pigmented centers, but it is unclear whether this represents simple co-precipitation of pigment with cholesterol during stone nidation or nidation on a true pigment stone center. To clarify this issue, we selected from among 67 sets of cholesterol gallstones, 12 sets with the most conspicuously pigmented centers. The composition of the centers and the peripheries of these 12 stones was analyzed using infrared spectroscopy and compared with that of 10 black pigment gallstones. The pigmented centers of cholesterol stones contained 80.1 +/- 7.9% (mean +/- S.E.) cholesterol, 6.2 +/- 3.4% calcium bilirubinate (only 4 of the 12 centers had measurable calcium bilirubinate), trace amounts of calcium phosphate and no calcium carbonate or calcium palmitate. The peripheral areas of the cholesterol stones contained 91.6 +/- 2.3% cholesterol and no detectable calcium salts. For comparison, the composition of the centers of 10 black pigment gallstones was 13.5 +/- 2.2% cholesterol, 28.2 +/- 5.3% calcium bilirubinate, 5.5 +/- 2.4% calcium phosphate and 10.6 +/- 5.8% calcium carbonate. The composition of only one cholesterol stone center (15.8% cholesterol, 26.8% calcium bilirubinate) resembled that of a pigment stone, but even this center differed from that of a typical pigment stone in that it contained only a trace amount of calcium phosphate and no calcium carbonate. Thus, the chemical composition of pigmented centers of cholesterol gallstones is quantitatively different from that of black pigment stones, suggesting that cholesterol gallstones do not form on a pigment stone nidus.


Real-time landslide warning during heavy rainfall.
A real-time system for issuing warnings of landslides during major storms is being developed for the San Francisco Bay region, California. The system is based on empirical and theoretical relations between rainfall and landslide initiation, geologic determination of areas susceptible to landslides, real-time monitoring of a regional network of telemetering rain gages, and National Weather Service precipitation forecasts. This system was used to issue warnings during the storms of 12 to 21 February 1986, which produced 800 millimeters of rainfall in the region. Although analysis after the storms suggests that modifications and additional development are needed, the system successfully predicted the times of major landslide events. It could be used as a prototype for systems in other landslide-prone regions.


IL-27p28 specifically regulates MHC II expression in macrophages through CIITA.
Antigen-presenting cells (APCs) constantly express major histocompatibility complex II (MHC II), including macrophages and dendritic cells (DCs) which deliver antigens to CD4+ T cells and play an important role in adaptive immunity. The expression of MHC II is controlled by the transcriptional coactivator CIITA. Interleukin-27 (IL-27), a newly discovered IL-12 family cytokine, is composed of p28 and EBI3 subunits. In this study, we used IL-27p28 conditional knock-out mice to investigate the regulatory effects of IL-27p28 on macrophage polarization and the expression of MHC II in macrophages. We found that MHC II expression was upregulated in the bone marrow-derived and peritoneal exudate macrophages (BMDMs; PEMs) from IL-27p28-deficient mice, with their inflammation regulating function unaffected. We also demonstrated that in the APCs, IL-27p28 selectively regulated MHC II expression in macrophages but not in dendritic cells. During Pseudomonas aeruginosa (P. aeruginosa) reinfection, higher survival rate, bacterial clearance, and ratio of CD4+/CD8+ T cells in the spleen during the specific immune phase were observed in IL-27p28 defect mice, as well as an increased MHC II expression in alveolar macrophages (AMs). But these did not occur in the first infection. For the first time we discovered that IL-27p28 specifically regulates the expression of MHC II in macrophages by regulating CIITA, while its absence enhances antigen presentation and adaptive immunity against P. aeruginosa.


The influence of mortality and socioeconomic status on risk and delayed rewards: a replication with British participants.
Here, we report three attempts to replicate a finding from an influential psychological study (Griskevicius et al., 2011b). The original study found interactions between childhood SES and experimental mortality-priming condition in predicting risk acceptance and delay discounting outcomes. The original study used US student samples. We used British university students (replication 1) and British online samples (replications 2 and 3) with a modified version of the original priming material, which was tailored to make it more credible to a British audience. We did not replicate the interaction between childhood SES and mortality-priming condition in any of our three experiments. The only consistent trend of note was an interaction between sex and priming condition for delay discounting. We note that psychological priming effects are considered fragile and often fail to replicate. Our failure to replicate the original finding could be due to demographic differences in study participants, alterations made to the prime, or other study limitations. However, it is also possible that the previously reported interaction is not a robust or generalizable finding.


Atomic resolution structure of obelin: soaking with calcium enhances electron density of the second oxygen atom substituted at the C2-position of coelenterazine.
The spatial structure of the Ca(2+)-regulated photoprotein obelin has been solved to resolution of 1.1A. Two oxygen atoms are revealed substituted at the C2-position of the coelenterazine in contrast to the obelin structure at 1.73A resolution where one oxygen atom only was disclosed. The electron density of the second oxygen atom was very weak but after exposing the crystals to a trace of Ca(2+), the electron densities of both oxygen atoms became equally intense. In addition, one Ca(2+) was found bound in the loop of the first EF-hand motif. Four of the ligands were provided by protein residues Asp30, Asn32, Asn34, and the main chain oxygen of Lys36. The other two were from water molecules. From a comparison of B-factors for the residues constituting the active site, it is suggested that the variable electron densities observed in various photoprotein structures could be attributed to different mobilities of the peroxy oxygen atoms.


Increased efficiency using the encapsulation-dehydration cryopreservation technique for Arabidopsis thaliana.
Arabidopsis thaliana shoot tips were successfully cryopreserved using encapsulation-dehydration cryopreservation methods. Between one and seven shoot tips were encapsulated within 4 mm calcium-alginate beads. Beads were formed in the presence of 2 M glycerol + 0.4 M sucrose. The time required to make 10 beads, each containing five shoot tips (4 min), was less than the time required to make 50 beads containing one shoot tip (12 min). Shoot tip regrowth after cryoexposure was between 60 and 68%, with one to seven shoot tips per bead. Using five Arabidopsis shoot tips per bead, alginate beads were formed either in the presence of 2 M glycerol + 0.4 M sucrose or 0.5 M sucrose. Beads formed in the presence of glycerol were immediately air-dried to moisture contents between 0.21 to 0.26 g H2O/g FW (0.27 to 0.38 g H2O/g DW). Alginate beads formed in 0.5 M sucrose were incubated in solutions of 0.5, 0.75, and 1 M sucrose for one day each prior to air-dehydration, achieving moisture contents of 0.19 to 0.21 g H2O/g FW (0.23 to 0.27 g H2O/g DW). Shoot tip regrowth after cryoexposure was between 42 and 65%, with no significant differences among treatments. We successfully reduced the amount of time needed for shoot tip processing for Arabidopsis by encapsulating five shoot tips per alginate bead and by using a glycerol-encapsulation method, without lowering shoot tip regrowth levels after cryopreservation.


Left ventricular mass regression after sutureless implantation of the Perceval S aortic valve bioprosthesis: preliminary results.
Left ventricular (LV) hypertrophy in aortic stenosis (AS) is considered a compensatory response helping maintain systolic function, but constitutes a risk factor for cardiac morbidity and mortality. The aim of this study was to assess the degree of LV mass regression after sutureless implantation of the Perceval S aortic valve bioprosthesis (Sorin Group, Saluggia, Italy). Between March 2010 and July 2012, 78 patients with symptomatic AS underwent isolated aortic valve replacement (AVR) with the Perceval bioprosthesis. Mean age was 77.1 ± 5.3 years, 46 patients were female (59%) and mean logistic EuroSCORE was 11 ± 7.5%. Echocardiography was performed preoperatively, at discharge, and at follow-up (mean 13.5 ± 7.3 months). LV mass was calculated using the Devereux formula and indexed to body surface area. There was 1 in-hospital non-cardiac death and 3 late deaths. LV mass index decreased from 148.4 ± 46 g/m(2) at baseline to 119.7 ± 38.5 g/m(2) at follow-up (P = 0.002). No significant changes were observed in LV hypertrophy and/or relative wall thickness >0.42 as well as in LV ejection fraction. Mean aortic gradient decreased from 49.5 ± 15.8 mmHg at baseline to 11.6 ± 5.1 mmHg at discharge and 8.3 ± 4.4 mmHg at follow-up (P < 0.001), resulting in significant clinical improvement. No moderate or severe paravalvular leakage was observed at discharge and at follow-up. In AS patients, isolated AVR with the Perceval sutureless bioprosthesis is associated with significant LV mass regression at 1-year follow-up. However, longer-term follow-up is necessary to confirm these findings.


Effects of spatial distribution of the nuclear localization sequence on gene transfection in catiomer-gene polyplexes.
A spatial effect of targeted signal distribution is found in catiomer-gene polyplexes which strongly influences the gene transfection profiles. The nuclear localization sequence PKKKRKV (NLS) is chosen as the target ligand. For a given gene delivery system of constant composition, size, and charge, it is engineered such that the NLS is the only structural variable in polyplexes. This unique architecture of polyplexes relies on successive electrostatic interaction. The structural features of the PKKKRKV sequence in polyplexes are investigated by NMR. The gene transfection profiles of the polyplexes are found to be well correlated with the spatial distribution of the nuclear localization sequence. Upon modification of nuclear localization, the gene transfection efficiency is found to increase remarkably from 40% to 60%. The drastic improvement of gene transfection is explained by a NLS spatial distribution mechanism. High gene transfection based on spatial NLS distribution provides a new base for the design and development of non-viral gene delivery vectors.


Improved survival with inhibitory killer immunoglobulin receptor (KIR) gene mismatches and KIR haplotype B donors after nonmyeloablative, HLA-haploidentical bone marrow transplantation.
Natural killer (NK) cell alloreactivity, which may contribute to the graft-versus-leukemia (GVL) effect of allogeneic hematopoietic stem cell transplantation (HSCT), is influenced by the interaction of killer-cell immunoglobulin-like receptors (KIRs) on donor NK cells and their ligands, human leukocyte antigen (HLA) class I molecules on recipient antigen-presenting cells (APCs). Distinct models to predict NK cell alloreactivity differ in their incorporation of information from typing of recipient and donor KIR and HLA gene loci, which exist on different autosomes and are inherited independently as haplotypes. Individuals may differ in the inheritance of the 2 KIR haplotypes, A and B, or in the expression of individual KIR genes. Here, we examined the effect of KIR and HLA genotype, in both the recipient and donor, on the outcome of 86 patients with advanced hematologic malignancies who received nonmyeloablative (NMA), HLA-haploidentical HSCT with high-dose, posttransplantation cyclophosphamide (Cy). Compared to recipients of bone marrow (BM) from donors with identical KIR gene content, recipients of inhibitory KIR (iKIR) gene-mismatched BM had an improved overall survival (OS) (hazard ratio [HR]=0.37; confidence interval [CI]: 0.21-0.63; P=.0003), event-free survival (EFS) (HR=0.51; CI: 0.31-0.84; P=.01), and relapse rate (cause-specific HR, SDHR=0.53; CI: 0.31-0.93; P=.025). Patients homozygous for the KIR "A" haplotype, which encodes only 1 activating KIR, had an improved OS (HR=0.30; CI: 0.13-10.69; P=.004), EFS (HR=0.47; CI: 0.22-1.00; P=.05), and nonrelapse mortality (NRM; cause-specific HR=0.13; CI: 0.017-0.968; P=.046) if their donor expressed at least 1 KIR B haplotype that encodes several activating KIRs. Models that incorporated information from recipient HLA typing, with or without donor HLA typing, were not predictive of outcome in this patient cohort. Thus, NMA conditioning and T cell-replete, HLA-haploidentical HSCTs involving iKIR gene mismatches between donor and recipient, or KIR haplotype AA recipients of BM from KIR Bx donors, were associated with lower relapse and NRM and improved OS and EFS. These findings suggest that selection of donors based upon inhibitory KIR gene or haplotype incompatibility may be warranted.


The Impact of Externally Worn Diabetes Technology on Sexual Behavior and Activity, Body Image, and Anxiety in Type 1 Diabetes.
We aimed to explore the impact of externally worn diabetes technologies on sexual behavior and activity, body image, and anxiety in adopters and nonadopters of these devices. People with type 1 diabetes aged 16-60 years living in Western Australia were invited to complete an online survey. Of the 289 respondents (mean age 34.3 years), 45% used continuous subcutaneous insulin infusion (CSII) and 35% used continuous glucose monitoring (CGM). Approximately half of CSII users stated that the pump interferes with sex. Of these, 75% disconnect their pump during sexual activity to avoid this issue. Comfort during sex influenced the location of the CSII insertion site in 22% of respondents, with the abdomen being preferred. One in four non-CSII users cited sex-related concerns as a factor for not adopting the technology. CGM interfered with sexual activity in 20% of users, but did not commonly affect CGM placement (only 18%). Sexual activity was reported as a factor for not adopting the technology in 10% of non-CGM users. No differences in body dissatisfaction (P = .514) or anxiety (P = .304) between CSII and non-CSII users were observed. No differences in sexual activity and behavior between technology users and nontechnology users were observed. Wearable technologies impact upon sexual activity and this influences the decision to adopt the technology. Despite this, technology users are similar in terms of sexual behavior, anxiety, and body image compared to nontechnology users. Where appropriate, these data can be used to identify potential concerns, address strategies to mitigate them, and inform people with diabetes when considering adopting external technologies.


Impact of Critical Care Point-of-Care Ultrasound Short-Courses on Trainee Competence.
Competence in point-of-care ultrasound is recommended/mandated by several critical care specialties. Although doctors commonly attend point-of-care ultrasound short-courses for introductory training, there is little follow-up data on whether they eventually attain competence. This study was done to determine the impact of point-of-care ultrasound short-courses on point-of-care ultrasound competence. Web-based survey. Follow-up after point-of-care ultrasound short-courses in the Asia-Pacific region. Doctors who attended a point-of-care ultrasound short-course between December 2015 and February 2018. Each subject was emailed a questionnaire on or after 6 months following their short-course. They were asked if they had performed at least 30 structured point-of-care ultrasound scans and/or reached point-of-care ultrasound competence and their perceived reasons/challenges/barriers. They were also asked if they used point-of-care ultrasound as a clinical diagnostic aid. The response rate was 74.9% (182/243). Among the 182 respondents, only 12 (6.6%) had attained competence in their chosen point-of-care ultrasound modality, attributing their success to self-motivation and time management. For the remaining doctors who did not attain competence (170/182, 93.4%), the common reasons were lack of time, change of priorities, and less commonly, difficulties in accessing an ultrasound machine/supervisor. Common suggestions to improve short-courses included requests for scanning practice on acutely ill ICU patients and prior information on the challenges regarding point-of-care ultrasound competence. Suggestions to improve competence pathways included regular supervision and protected learning time. All 12 credentialled doctors regularly used point-of-care ultrasound as a clinical diagnostic aid. Of the 170 noncredentialled doctors, 123 (72.4%) reported performing unsupervised point-of-care ultrasound for clinical management, either sporadically (42/170, 24.7%) or regularly (81/170, 47.7%). In this survey of doctors attending point-of-care ultrasound short-courses in Australasia, the majority of doctors did not attain competence. However, the practice of unsupervised point-of-care ultrasound use by noncredentialled doctors was common. Further research into effective strategies to improve point-of-care ultrasound competence is required.


[Sugar free diet: a new perspective in the treatment of Crohn disease? Randomized, control study].
Since several studies have shown that patients with Crohn's disease have an increased consumption of refined carbohydrates, the influence of a diet excluding refined sugar on the course of the disease was examined. In a randomised control trial, 20 patients (10 patients in each group) with Crohn's disease were treated for an average of 18 months with two different diets. The patients used in the study had a low or middle activity of the disease. Drug treatment was omitted 14 days before commencement of the study. The first group was treated with a low carbohydrate diet (refined sugar excluded), the second group received a high carbohydrate diet (refined sugar-rich). In patients with higher activities of the disease (activity index 100-200 points), the diet which restricted refined sugar was superior to the sugar-rich diet; in 4 out of 5 patients the disease activity decreased and remained so throughout the study-period. In contrast to this 4 patients treated with the sugar-rich diet had to be taken off the treatment because of increasing activities of the disease. In patients with quiescent disease (activity index less than 100 points), neither of the diets showed detrimental effects. The statistical analysis of clinical and laboratory dates noted during the study period resulted in no significant differences between the two groups.


Efficacy of the lateral advanced flap in root-coverage procedures for mandibular central incisors: a 5-year clinical study.
The aim of this study was to determine the efficacy of the lateral advanced flap in root-coverage procedures for mandibular incisors and to evaluate pain after treatment. A total of 15 patients who required treatment for gingival recessions were selected from the University of Genoa Laser and Restorative Dentistry Department. The inclusion criteria were the presence of at least one 3-mm gingival recession defect of a mandibular incisor and 3 mm or more of keratinized tissue width on the immediately adjacent tooth. One calibrated masked examiner performed the clinical measurements, including recession depth and width, probing depth, clinical attachment level, and keratinized tissue width. Patients were checked 7, 14, and 30 days after surgery and were included in supportive periodontal maintenance every 4 months. Further follow-ups were done at the first, third, and fifth year postsurgery when the same baseline assessment parameters were recorded according to the root-coverage esthetic score (RES). Numeric score recorded pain evaluations showed a mean of 3 at the day of the surgery, 2 in the first day after the surgery, and no significant scores were referred in the following days. Mean recession depth was 3.2 ± 1.3 mm at baseline; at 5 years, RES showed a positive increase of all scores. The laterally coronally advanced surgical technique was very effective in treating isolated gingival recessions. It combined the esthetic and root-coverage advantages of the coronally advanced flap with the increased gingival thickness and keratinized tissue associated with the lateral gingival flap.


Escape mechanisms in tumor immunity: a year 2000 update.
The current consensus of opinion has it that most or possibly all tumors, spontaneous as well as induced, are immunogenic, expressing antigens in a form recognizable by the host immune system. Accordingly, in order to progress, tumors have to evolve strategies for evading immune responses. The purpose of this review is to consider the current status of knowledge concerning these different tumor escape strategies. It represents an update of an article originally published in this journal in 1997 (Pawelec, Zeuthen, and Kiessling, 1997). Therefore, it focuses mostly on publications that have appeared since then, illustrating the impressive accumulation of new data since that time and the importance currently attributed to studies of tumor escape from the immune response.


Force-fluorescence spectroscopy at the single-molecule level.
During the past decade, various powerful single-molecule techniques have evolved and helped to address important questions in life sciences. Yet these techniques would be even more powerful if they would be combined, that is, single-molecule manipulation with an orthogonal single-molecule observation. Here, we present a recently developed approach to combine single-molecule optical tweezers with single-molecule fluorescence spectroscopy. Optical tweezers are used to manipulate and observe mechanical properties on the nanometer scale and piconewton force range. However, once the force range is in the low piconewton range or less, the spatial resolution of optical tweezers decreases significantly. In combination with fluorescence spectroscopy, like Förster resonance energy transfer (FRET), we are able to observe nanometer fluctuations and internal conformational changes in a low-force regime. The possibility to place fluorescent labels at nearly any desired position and a sophisticated design of the experiment increases the amount of information that can be extracted in contrast to pure mechanical or fluorescence experiments.


OmniMapFree: a unified tool to visualise and explore sequenced genomes.
Acquiring and exploring whole genome sequence information for a species under investigation is now a routine experimental approach. On most genome browsers, typically, only the DNA sequence, EST support, motif search results, and GO annotations are displayed. However, for many species, a growing volume of additional experimental information is available but this is rarely searchable within the landscape of the entire genome. • We have developed a generic software which permits users to view a single genome in entirety either within its chromosome or supercontig context within a single window. This software permits the genome to be displayed at any scales and with any features. Different data types and data sets are displayed onto the genome, which have been acquired from other types of studies including classical genetics, forward and reverse genetics, transcriptomics, proteomics and improved annotation from alternative sources. In each display, different types of information can be overlapped, then retrieved in the desired combinations and scales and used in follow up analyses. The displays generated are of publication quality. • OmniMapFree provides a unified, versatile and easy-to-use software tool for studying a single genome in association with all the other datasets and data types available for the organism.


Improving pore interconnectivity in polymeric scaffolds for tissue engineering.
A new scaffold fabrication technique aiming to enhance pore interconnectivity for tissue engineering has been developed. Medical grade poly(lactic acid) was utilized to generate scaffolds by a solvent-evaporating/particulate-leaching technique, using a new dual-porogen system. Water-soluble sodium chloride particles were used to control macro-pore size in the range 106-255 microm, while organic naphthalene was utilized as a porogen to increase pore interconnections. The three-dimensional (3D) morphology of the scaffolds manufactured with and without naphthalene was examined by optical coherence tomography and scanning electron microscopy. The mechanical properties of the scaffolds were characterized by compression tests. MG63 osteoblast cells were seeded in the scaffolds to study the cell attachment and viability evaluated by confocal microscopy. It was revealed that introducing naphthalene as the second porogen in the solvent-evaporating/particulate-leaching process resulted in improvement of the pore interconnectivity. Cells grew in both scaffolds fabricated with and without naphthalene. They exhibited strong green fluorescence when using a live/dead fluorescent dye kit, indicating that the naphthalene in the scaffold process did not affect cell viability.


Gene therapy for carcinoma of the breast: Genetic ablation strategies.
The gene therapy strategy of mutation compensation is designed to rectify the molecular lesions that are etiologic for neoplastic transformation. For dominant oncogenes, such approaches involve the functional knockout of the dysregulated cellular control pathways provoked by the overexpressed oncoprotein. On this basis, molecular interventions may be targeted to the transcriptional level of expression, via antisense or ribozymes, or post-transcriptionally, via intracellular single chain antibodies (intrabodies). For carcinoma of the breast, these approaches have been applied in the context of the disease linked oncogenes erbB-2 and cyclin D1, as well as the estrogen receptor. Neoplastic revision accomplished in modal systems has rationalized human trials on this basis.


Phase I dose escalation study of KOS-1584, a novel epothilone, in patients with advanced solid tumors.
First-in-man study of KOS-1584, a second generation epothilone. Patients with advanced solid malignancies received KOS-1584 every 3 weeks until disease progression. Using a modified Fibonacci dose escalation scheme, one patient was enrolled at each dose level until the first instance of grade 2 toxicity. Thereafter, a standard 3 + 3 design was utilized. Sixty-six patients in 14 cohorts were dosed from 0.8 to 48 mg/m(2). Diarrhea, arthralgias, and encephalopathy were dose-limiting toxicities (DLTs) at doses ≥36 mg/m(2). At the recommended phase II dose (RP2D), the most common adverse effects were peripheral neuropathy (low grade), fatigue, arthralgias/myalgias, and diarrhea (31, 6%). The incidence of neutropenia was low. The overall clearance, volume of distribution, and half-life of KOS-1584 were 11 ± 6.17 L/h/m(2), 327 ± 161 L/m(2), and 21.9 ± 8.75 h, respectively. The half-life for the seco-metabolite (KOS-1891) was 29.6 ± 13.8 h. KOS-1584 exhibited linear pharmacokinetics. A dose-dependent increase in microtubulin bundle formation was observed at doses ≥27 mg/m(2). Two patients achieved partial responses and 24 patients had stable disease (SD). The RP2D of KOS-1584 is 36 mg/m(2). The lack of severe neurologic toxicity, diarrhea, neutropenia, or hypersensitivity reactions; favorable pharmacokinetic profile; and early evidence of activity support further evaluation.


Prognostic value of histological subtype in intraductal papillary mucinous neoplasm of the pancreas: A retrospective analysis of outcome from one single center.
We sought to retrospectively analyze the outcomes of patients with intraductal papillary mucinous neoplasm (IPMN) at our pancreatic surgery center, and to evaluate the prognostic value of histological subtype.The clinical data of 121 IPMNs treated in our center between 2005 and 2014 were retrospectively analyzed. Pathological slides were thoroughly reviewed by 2 specialized pathologists.Of the 121 patients, 48, 57, and 16 had main-duct, branch-duct, and mixed type IPMNs, respectively. Forty-one patients had invasive IPMNs. Histological subtypes consisted of 35 intestinal (28.9%), 56 gastric (46.3%), 29 pancreatobiliary (24.0%), and 1 oncocytic type (0.8%). Histological subtype was associated with radiological type, T stage, and degree of dysplasia (P < .05). No significant difference in overall survival was observed among the 4 histological subtypes, regardless of whether we considered all IPMNs (P = .106), or invasive IPMNs only (P = .828). However, the overall survival was associated with radiological type, T stage, degree of dysplasia, lymph-node status, and nerve invasion. For invasive IPMNs, the overall survival was associated with nerve invasion and lymph-node status; however, the association between nerve invasion and overall survival lost statistical significance after multivariate analysis.Histological subtype had limited prognostic value in patients with IPMNs, and the main prognostic factor for patients with invasive IPMNs was the lymph-node status.


Cost effectiveness of HMG-CoA reductase inhibition in Canada.
To assess the cost effectiveness of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor therapy, particularly atorvastatin, in primary and secondary prevention of coronary artery disease (CAD) in Canada. A Markov model was developed in which costs and effectiveness of atorvastatin were compared with those of other statins and with no drug therapy in primary and secondary prevention of CAD. Cost effectiveness was assessed for cohorts of patients with risk profiles defined by CAD status, age, sex, pretreatment low density lipoprotein cholesterol level and presence of sentinel coronary risk factors. Coronary risk was estimated by using initial and subsequent event coronary risk equations from the Framingham Heart Study, and risk factors were estimated by using Canadian population survey data. Recent estimates of the costs of CAD-related medical care in Canada were used to assign costs to health states and acute coronary events. Interventions included atorvastatin 10 mg, simvastatin 10 mg, pravastatin 20 mg, fluvastatin 20 mg, lovastatin 20 mg and no pharmacological therapy. Incremental cost effectiveness ratios (CDN$/year of life gained) relative to no therapy were lowest for atorvastatin and highest for pravastatin across all risk profiles. Atorvastatin was less costly and more effective than lovastatin, pravastatin and simvastatin in primary and secondary prevention, and conferred additional health benefits at a reduced cost per year of life gained compared with fluvastatin. Atorvastatin was found to be the most cost effective statin in primary and secondary prevention of CAD.


ATP-Sensitive Potassium Channels in Migraine: Translational Findings and Therapeutic Potential.
Globally, migraine is a leading cause of disability with a huge impact on both the work and private life of affected persons. To overcome the societal migraine burden, better treatment options are needed. Increasing evidence suggests that ATP-sensitive potassium (KATP) channels are involved in migraine pathophysiology. These channels are essential both in blood glucose regulation and cardiovascular homeostasis. Experimental infusion of the KATP channel opener levcromakalim to healthy volunteers and migraine patients induced headache and migraine attacks in 82-100% of participants. Thus, this is the most potent trigger of headache and migraine identified to date. Levcromakalim likely induces migraine via dilation of cranial arteries. However, other neuronal mechanisms are also proposed. Here, basic KATP channel distribution, physiology, and pharmacology are reviewed followed by thorough review of clinical and preclinical research on KATP channel involvement in migraine. KATP channel opening and blocking have been studied in a range of preclinical migraine models and, within recent years, strong evidence on the importance of their opening in migraine has been provided from human studies. Despite major advances, translational difficulties exist regarding the possible anti-migraine efficacy of KATP channel blockage. These are due to significant species differences in the potency and specificity of pharmacological tools targeting the various KATP channel subtypes.


The place of 'active birth' in present-day obstetric practice.
"Active birth' is becoming an increasingly prominent issue in obstetric practice. The psychosocial background of current public interest in the subject is examined. Guidelines are suggested to accommodate "active-birth' practices within the hospital environment. The Johannesburg Hospital experience of the application of these guidelines in the first 100 patients admitted to the "Midwives Active Birth Unit' is reported.


CNS-targeted production of IL-17A induces glial activation, microvascular pathology and enhances the neuroinflammatory response to systemic endotoxemia.
Interleukin-17A (IL-17A) is a key cytokine modulating the course of inflammatory diseases. Whereas effector functions of IL-17A like induction of antimicrobial peptides and leukocyte infiltration could clearly be demonstrated for peripheral organs, CNS specific effects are not well defined and appear controversial. To further clarify the functional significance of IL-17A in the CNS, we generated a transgenic mouse line with astrocyte-restricted expression of the IL-17A gene. GFAP/IL-17A transgenic mice develop normally and do not show any signs of neurological dysfunction. However, histological characterization revealed astrocytosis and activation of microglia. Demyelination, neurodegeneration or prominent tissue damage was not observed but a vascular pathology mimicking microangiopathic features was evident. Histological and flow cytometric analysis demonstrated the absence of parenchymal infiltration of immune cells into the CNS of GFAP/IL-17A transgenic mice. In GFAP/IL-17A mice, LPS-induced endotoxemia led to a more pronounced microglial activation with expansion of a distinct CD45(high)/CD11b(+) population and increased induction of proinflammatory cytokines compared with controls. Our data argues against a direct role of IL-17A in mediating tissue damage during neuroinflammation. More likely IL-17A acts as a modulating factor in the network of induced cytokines. This novel mouse model will be a very useful tool to further characterize the role of IL-17A in neuroinflammatory disease models.


[Magnetic resonance tomography with endorectal coil for examination of prostate and seminal vesicles].
MRI with an endorectal coil gives images of the prostate gland and seminal vesicles of higher quality than any other imaging modality. The use of an endorectal coil enables us to sample stronger radiofrequency signals from the prostate gland and seminal vesicles compared to a whole-body coil. This improves the contrast resolution in MR images and may give a higher spatial resolution by use of thin slices and a smaller slice gap. Intravenous MRI contrast is not used in our standard procedure. At Haukeland University Hospital, Norway, MRI with an endorectal coil is presently used as a tool for solving special problems in relation to detection and staging of prostate cancer, for example in patients with an abnormal s-PSA and negative sextant biopsies from the prostate gland. Furthermore, MRI with an endorectal coil may be used for the diagnosis of some benign diseases. The specific role of MRI with an endorectal coil in clinical use is not clarified; further research is needed. This article gives a short overview of technical aspects, imaging protocols, image reading, indications and diagnostic criteria for some diseases.


Vectorcardiographic loop alignment for fetal movement detection using the expectation-maximization algorithm and support vector machines.
Reduced fetal movement is an important parameter to assess fetal distress. Currently, no suitable methods are available that can objectively assess fetal movement during pregnancy. Fetal vectorcardiographic (VCG) loop alignment could be such a method. In general, the goal of VCG loop alignment is to correct for motion-induced changes in the VCGs of (multiple) consecutive heartbeats. However, the parameters used for loop alignment also provide information to assess fetal movement. Unfortunately, current methods for VCG loop alignment are not robust against low-quality VCG signals. In this paper, a more robust method for VCG loop alignment is developed that includes a priori information on the loop alignment, yielding a maximum a posteriori loop alignment. Classification, based on movement parameters extracted from the alignment, is subsequently performed using support vector machines, resulting in correct classification of (absence of) fetal movement in about 75% of cases. After additional validation and optimization, this method can possibly be employed for continuous fetal movement monitoring.


Kikuchi disease: differentiation from tuberculous lymphadenitis based on patterns of nodal necrosis on CT.
KD, also known as histiocytic necrotizing lymphadenitis, is often mimicked by TL, especially when KD is accompanied by gross necrosis on CT. We analyzed the nodal necrosis patterns on CT in patients with KD and TL to identify the predictive features that differentiate these 2 entities. We included 24 patients with KD (14 females, 10 males; mean age, 25.1 years) and 45 with TL (36 women, 9 men; mean age, 39.9 years) having nodal necrosis on CT. The extent, number, location, and margins of nodal necrosis; the presence of perinodal infiltration; and nodal calcification were analyzed. Two attenuation indices for nodal necrosis were derived: CTN and CTN/M. The CT features of the 2 diseases were compared, and multivariate logistic regression analysis was performed to identify predictive features that would aid in differentiating KD from TL. Mild or moderate extent of nodal necrosis (P = .000), multiple necrotic foci (P = .002), indistinct margins of necrotic foci (P = .000), CTN of >44.5 (P = .000), CTN/M of >0.7 (P = .000), and absence of nodal calcification (P = .012) were associated with KD. Multivariate logistic regression analysis revealed that indistinct margins of necrotic foci were independent predictors, with 80% accuracy. The presence of indistinct margins of necrotic foci was an independent predictor in differentiating KD from TL with high diagnostic accuracy.


Uridine Diphosphate-Glucuronosyltransferase (UGT) 2B Subfamily Interspecies Differences in Carnivores.
UDP-glucuronosyltransferases (UGTs) are among the most important xenobiotic metabolizing enzymes that conjugate a wide range of chemicals. Previous studies showed that Felidae and Pinnipedia species have very low UGT activities toward some phenolic compounds because of the UGT1A6 pseudogene and small numbers of UGT1A isozymes. In addition to the UGT1As, UGT2Bs isozymes also conjugate various endogenous (eg, estrogens, androgens, and bile acids) and exogenous compounds (opioids, non-steroidal anti-inflammatory drugs, and environmental pollutants). However UGT2B activity and genetic background are unknown in carnivore species. Therefore, this study was performed to elucidate the species differences of UGT2Bs. Using typical substrates for UGT2Bs, UGT activity was measured in vitro. In addition, UGT2B genetic features are analyzed in silico. Results of UGT activity measurement indicate marked species differences between dogs and other carnivores (cats, Northern fur seals, Steller sea lions, Harbor seals, and Caspian seals). Dogs have very high Vmax/Km toward estradiol (17-glucuronide), estrone, lorazepam, oxazepam, and temazepam. Conversely, cats and pinniped species (especially Caspian seals and Harbor seals) have very low activities toward these substrates. The results of genetic synteny analysis indicate that Felidae and pinniped species have very small numbers of UGT2B isozymes (one or none) compared with dogs, rodents, and humans. Furthermore, Felidae species have the same nonsense mutation in UGT2B, which suggests that Felidae UGT2B31-like is also a pseudogene in addition to UGT1A6. These findings of lower activity of UGT2B suggest that Felidae and some pinniped species have very low UGT activity toward a wide range of chemicals. These results are important for Felidae and Pinnipedia species that are frequently exposed to drugs and environmental pollutants.


Thiyl radical promoted iron-catalyzed-selective oxidation of benzylic sp3 C-H bonds with molecular oxygen.
A ligand-free iron-catalyzed method for the oxygenation of benzylic sp3 C-H bonds by molecular oxygen (1 atm) using a thiyl radical as a cocatalyst has been developed. This transformation provides a facile access to amides, esters and ketones from readily accessible corresponding amines, ethers and alkanes. It features high regioselectivity, mild oxidative conditions and excellent functional group compatibility, providing good opportunities to the site-selective functionalization of complex molecules. Preliminary mechanistic studies suggest that this reaction may not undergo a benzylic cation intermediate pathway and the carbonyl oxygen atom in the products may be derived from molecular oxygen.


CCR1/CCL5 (RANTES) receptor-ligand interactions modulate allogeneic T-cell responses and graft-versus-host disease following stem-cell transplantation.
Acute graft-versus-host disease (GVHD) and leukemic relapse are serious complications of allogeneic stem-cell transplantation (SCT). Recruitment of activated T cells to host target tissues or sites of leukemic infiltration (graft-versus-leukemia [GVL]) is likely mediated by chemokine receptor-ligand interactions. We examined the contribution of donor cell CCR1 expression to the development of GVHD and GVL using a well-established murine SCT model (B6 --> B6D2F1) and CCR1-deficient mice (CCR1(-/-)). Allo-SCT with CCR1(-/-) donor cells significantly reduced systemic and target organ GVHD severity, and CCR1 expression on both T cells and accessory cells contributed to GVHD mortality. Significant GVL activity was preserved following CCR1(-/-) SCT, but the survival advantage diminished with increasing tumor burden. We then explored the effects of CCR1 expression on allo-specific T-cell responses. Although cytolytic effector function was maintained on a per-cell basis, T-cell proliferation and IFNgamma secretion were significantly reduced both in vivo and in vitro. T-cell function was partially dependent on interactions between CCR1 and CCL5. Collectively, these data demonstrate that CCR1 expression on donor cells contributes to the development of both GVHD and GVL, and suggest that CCR1/CCL5 receptor-ligand interactions modulate allo-specific T-cell responses occurring in this context.


Bacterial Carotenoids: Extraction, Characterization, and Applications.
Natural carotenoids are secondary metabolites that exhibit antioxidant, anti-inflammatory, and anti-cancer properties. These types of compounds are highly demanded by pharmaceutical, cosmetic, nutraceutical, and food industries, leading to the search for new natural sources of carotenoids. In recent years, the production of carotenoids from bacteria has become of great interest for industrial applications. In addition to carotenoids with C40-skeletons, some bacteria have the ability to synthesize characteristic carotenoids with C30-skeletons. In this regard, a great variety of methodologies for the extraction and identification of bacterial carotenoids has been reported and this is the first review that condenses most of this information. To understand the diversity of carotenoids from bacteria, we present their biosynthetic origin in order to focus on the methodologies employed in their extraction and characterization. Special emphasis has been made on high-performance liquid chromatography-mass spectrometry (HPLC-MS) for the analysis and identification of bacterial carotenoids. We end up this review showing their potential commercial use. This review is proposed as a guide for the identification of these metabolites, which are frequently reported in new bacteria strains.


Managing the elephant in the room: responding to COVID-19's impact on data collection.
The global COVID-19 pandemic has affected data collection for many researchers, in particular research that involves face-to-face interviews. To share learning about the challenges encountered when face-to-face interviews could not continue as planned in a study and how to adapt data collection so that it can continue despite severe disruption. This article examines the considerations and actions taken by the authors to ensure the continuity of data collection. The research aimed to use narrative inquiry to understand the experiences of significant others supporting people in intensive care units with severe burn injuries. But the pandemic meant the researchers could not meet face-to-face with participants as originally intended and so they had to consider new ways to approach data collection. The authors explore the process of adapting the interviews to video conferencing and telephone use while preserving the study's person-centred focus to remain coherent with narrative methodology. Adapting data collection is valuable in ensuring the continuity of research. Careful consideration and planning are required to ensure the research remains robust and ethically sound. Adapting data collection methods can allow for greater flexibility when participants cannot attend face-to-face interviews.


Executive Function Skills Are Linked to Restricted and Repetitive Behaviors: Three Correlational Meta Analyses.
There is a consensus on the centrality of restricted and repetitive behaviors (RRBs) in the diagnosis of Autism Spectrum Disorder (ASD), yet the origins of these behaviors are still debated. We reconsider whether executive function (EF) accounts of RRBs should be revisited. EF deficits and high levels of RRBs are often pronounced in individuals with ASD and are also prevalent in young typically developing children. Despite this, the evidence is mixed, and there has been no systematic attempt to evaluate the relationship across studies and between task batteries. We examine recent evidence, and in three highly powered random-effects analyses (N = 2964), examine the strength of the association between RRB levels and performance on set shifting, inhibitory control, and parental-report based EF batteries. The analyses confirm significant associations between high levels of the behaviors and poor EF skills. Moreover, the associations remained stable across typical development and in individuals with ASD and across different types of EF measures. These meta-analyses consolidate recent evidence identifying that cognitive mechanisms correlate with high RRBs that are seen in individuals with ASD, as well as in typical development. We propose that the EF account may be critical for guiding future interventions in ASD research. LAY SUMMARY: Restricted and repetitive behaviors (RRBs) are diagnostic criteria for Autism yet also common in typical development, and if they persist over time some can have a negative impact on learning and social acceptance. The present meta-analyses found that high levels of RRBs related to poor performance on set-shifting and inhibitory control tasks, as well as high ratings on parental report scales. Future studies should create interventions that aim to improve these skills as they may help manage challenging RRBs.


Relationship between lipid peroxidation and disease activity in patients with Behçet's disease.
Behçet's disease is a chronic multi-systemic disorder which is characterized by a relapsing systemic inflammatory process. The alteration of lipid profile and lipid peroxidation resulting from the inflammatory process may he associated with an increased risk for atherosclerosis in patients with Behcet's disease. We investigated lipids, lipoprotein and lipid peroxidation and their inter-relationships considering the disease activity. Eighteen patients (11 male and 7 female) and 20 age-matched healthy subjects (10 male and 10 female) were studied. Lipid profile including total cholesterol, triacylglycerol, HDL-cholesterol, LDL-cholesterol, lipoprotein(a), apoAI and apoB, and acute phase reactants including polymorphonuclear (PMN) elastase, PMN leukocyte count, erythrocyte sedimentation rate (ESR) and complements (C3, C4) were evaluated in patients in active and inactive periods of Behçet's disease and control subjects. The levels of thiobarbituric acid reactive substance (TBARS) were assessed as an indicator of lipid peroxidation. Lipid peroxidation was found to he increased in the active period compared to the inactive period of the disease and control subjects. Also, lipid peroxidation showed correlations of various degrees with atherogenic lipid parameters in both periods of the followed-up patients. In conclusion, patients with Behçet's disease in the active period may be much more susceptible to atherogenic events than those in the inactive period of the disease and control subjects.


Muscle Strength and Changes in Physical Function in Women With Systemic Lupus Erythematosus.
Cross-sectional studies have observed that muscle weakness is associated with worse physical function among women with systemic lupus erythematosus (SLE). The present study examines whether reduced upper and lower extremity muscle strength predict declines in function over time among adult women with SLE. One hundred forty-six women from a longitudinal SLE cohort participated in the study. All measures were collected during in-person research visits approximately 2 years apart. Upper extremity muscle strength was assessed by grip strength. Lower extremity muscle strength was assessed by peak knee torque of extension and flexion. Physical function was assessed using the Short Physical Performance Battery (SPPB). Regression analyses modeled associations of baseline upper and lower extremity muscle strength with followup SPPB scores controlling for baseline SPPB, age, SLE duration, SLE disease activity (Systemic Lupus Activity Questionnaire), physical activity level, prednisone use, body composition, and depression. Secondary analyses tested whether associations of baseline muscle strength with followup in SPPB scores differed between intervals of varying baseline muscle strength. Lower extremity muscle strength strongly predicted changes over 2 years in physical function even when controlling for covariates. The association of reduced lower extremity muscle strength with reduced physical function in the future was greatest among the weakest women. Reduced lower extremity muscle strength predicted clinically significant declines in physical function, especially among the weakest women. Future studies should test whether therapies that promote preservation of lower extremity muscle strength may prevent declines in function among women with SLE.


Cationic surfactants for micellar electrokinetic chromatography: 2. Representative applications to acidic, basic, and hydrophobic analytes.
Changes in MEKC chemical selectivity that are induced by changes in the headgroup structure of cationic surfactants are examined. Separations of acidic, basic, and hydrophobic solutes are examined. The acidic analytes are comprised of methoxyphenols, which are of interest due to their prevalence in wood smoke. The basic solutes consist of compounds often found in forensic urine analysis, and represent typical basic pharmaceuticals. The hydrophobic solutes are six pharmaceutical corticosteroids used in replacement therapy of adrenocortical insufficiency and nonspecific treatment of inflammatory and allergic conditions. The role of the headgroup was found to be quite significant when analyzing acidic compounds with not all the surfactants being able to resolve all of the analytes. The headgroup also induced migration order switches among the acidic analytes. All of the surfactants examined here in were found to be suitable for the analysis of basic analytes with each surfactant providing unique selectivity. The hydrophobic solutes were separated best with the larger more hydrophobic surfactant headgroups. The steroid separation with these two surfactants was achieved without the use of organic modifiers or a mixed micellar phase.


Production of xylitol from Candida tropicalis by using an oxidation-reduction potential-stat controlled fermentation.
An on-line device, ORP (oxidation-reduction potential)-stat, was used to control glucose-feeding for enhancing xylitol conversion from D-xylose during an oxygen-limited fermentation by Candida tropicalis. The fermentation was carried out in a 5 l jar fermenter. After glucose in the medium was depleted, a switching to a limited aeration and feeding glucose controlled by ORP-stat was performed. The maximum xylitol yield was obtained under a condition at an ORP of - 180 mV and at an aeration rate of 0.2 l min(-1).


[Efficiency of ursodeoxycholic acid therapy in non-alcoholic fatty liver disease associated with metabolic syndrome].
Nonalcoholic Fatty Liver Disease (NAFLD) is one of the most common liver disease. Its prevalence is 20-30% in the population of developed countries, its prevalence is 26% in Russia. NAFLD is observed in many patients with Metabolic Syndrome. Because of the wide prevalence of this disease it is required to find best drugs influencing mechanisms of its development, chronicity and progression. 36 patients were included to the study. Mean age of patients was 43 +/- 3.9 years. Patients of the main group received the ursodeoxycholic acid within 2 months. Patients of the control group received the plant origin hepatoprotective medicine. All patients before and after treatment were carried out the biochemical analysis of blood, the liver ultrasound examination, the bioimpedance body composition analysis, the microbiological examination of faeces, the examination of metabolites of microorganisms in the blood by the method of gas-liquid chromatography - mass spectrometry, developed by Osipov G. A. The reduction of hepatic transaminases, the trend to normalization of the lipid profile, the reduction of body weight, the reduction of amount of adipose tissue in the body, the increase of Bifidobacterium spp., Lactobacillus spp., Enterococcus spp., the increase of levels of Bifidobacterium spp., Lactobacillus spp. Metabolites, the decrease of endotoxin plasma level and the decrease of total microbial load were observed after the UDCA treatment. The results of the study showed the prospectivity of the using of UDCA for NAFLD associated with Metabolic Syndrome.


Histone ubiquitination and chromatin remodeling in mouse spermatogenesis.
Male infertility in HR6B knockout mice is associated with impairment of spermatogenesis. The HR6B gene is a mammalian, autosomal homolog of the Saccharomyces cerevisiae gene Rad6 encoding a ubiquitin-conjugating enzyme. In addition, X-chromosomal HR6A has been identified, in human and mouse. RAD6 in yeast is required for a variety of cellular functions, including sporulation, DNA repair, and mutagenesis. Since RAD6 and its mammalian homologs can ubiquitinate histones in vitro, we have investigated the pattern of histone ubiquitination in mouse testis. By immunoblot and immunohistochemical analysis of wild-type mouse testis, a high amount of ubiquitinated H2A (uH2A) was detected in pachytene spermatocytes. This signal became undetectable in round spermatids, but then increased again during a relatively short developmental period, in elongating spermatids. No other ubiquitinated histones were observed. In the HR6B knockout mice, we failed to detect an overt defect in the overall pattern of histone ubiquitination. For somatic cell types, it has been shown that histone ubiquitination is associated with destabilization of nucleosomes, in relation to active gene transcription. Unexpectedly, the most intense uH2A signal in pachytene spermatocytes was detected in the sex body, an inactive nuclear structure that contains the heterochromatic X and Y chromosomes. The postmeiotic uH2A immunoexpression in elongating spermatids indicates that nucleosome destabilization induced by histone ubiquitination may play a facilitating role during histone-to-protamine replacement.


An Economic Comparison of Treatment Strategies with Anakinra in Systemic Juvenile Idiopathic Arthritis (sJIA).
Systemic juvenile idiopathic arthritis (sJIA) is a rare, complex autoinflammatory disease with substantial morbidity, often characterized by fever, rash, and muscle pain, amongst other symptoms. Biologic agents, such as anakinra, have been successfully used to treat patients internationally, but their usage in some regions is limited to patients that have failed to achieve clinically inactive disease with corticosteroids and conventional synthetic disease-modifying anti-rheumatic drugs (csDMARDs). Use of anakinra early in the disease course leads to better clinical outcomes; however, longer-term costs for this treatment strategy have not been established. This study compares the economic implications of first-line versus later-line availability of anakinra for patients with sJIA. Data for patients treated with first-line anakinra were identified from a single-center, prospective study and compared to a combination of published trial and economic evaluation information to facilitate a comparison to later-line anakinra (ie, following corticosteroids + csDMARDs). Costs were estimated for product acquisition and medical resource utilization (MRU), including planned outpatient visits and unplanned hospital admissions. Total costs over a 5-year horizon were compared. Total 5-year product acquisition cost for the first-line anakinra strategy was €24,021, and for later-line anakinra was €20,471. The corresponding MRU costs were €19,197 (first-line) versus €25,425 (later-line). Overall 5-year costs (product acquisition and MRU) were lower for the first-line strategy (€43,218 versus €45,896). The use of anakinra for patients with sJIA in the first-line setting is efficacious to induce and sustain inactive disease, and the findings of this study show that this treatment strategy leads to cost savings through reduced medical expenditure.


Dollars and sense: the financial impact of Canadian wellness initiatives†.
The popular press reports anecdotal benefits of organizational initiatives that are designed to improve employees' work-life balance and wellness, but the long-term impact of these initiatives on firms' financial performance is unknown. Our longitudinal study of publicly traded Canadian organizations uses the strategic human resources management framework to explain why these initiatives may affect their financial performance. We use an exploratory factor analysis of three waves of archival data to identify two types of initiatives and regression to measure their impact on return on assets (ROA). Our findings suggest that, after 2 years, health promotion initiatives significantly decreased ROA, while work-life balance initiatives significantly increased it. Both effects became non-significant over the longer term.


Period Analysis of Intraracial Differences in Incidence and Survival Rates in Epithelial Ovarian Cancer.
To explain the difference in the incidence and relative survival in a population-based cohort of women with epithelial ovarian cancer (EOC) postdiagnosis in the last forty years. EOC is the most common type of all ovarian cancers, but there is inadequate information about the variations related to long-term EOC survival. We acquired the incidence and relative survival rate data from the Surveillance, Epidemiology, and End Results (SEER) registries to analyze the epidemiological variations from 1974 to 2013 in EOC-affected individuals. The survival disparities in EOC-specific individuals due to age, race, and socioeconomic status (SES) were performed by Kaplan-Meier analysis. The Results. The overall incidence of EOC progressively declined to 9.0 per 100,000 from 11.4 in the last forty years. The median survival rate improved to 48 months in the first decade from a previous of 27 months in the fourth decade. The 5-year relative survival rate (RSR) increased to 44.3% that was previously 32.3% at the same time. However, between whites and blacks, an increase from 11 to 18 months was observed in the median survival differences. Between the low and high poverty groups, it was increased from 7 months to 12 months, respectively. The incidence rate of RSR and EOC-specific individuals in the last forty years was improved. However, the survival rates among different races and SES differed over time.


Calcium channel stability measured by gradual loss of excitability in pawn mutants of Paramecium aurelia.
Mutants of Paramecium aurelia that are unable to reverse swimming direction are called pawns. They lack the inward ionic (calcium) current required for the upstroke of the electrically excitable membrane response. By following the progressive loss of reversal response and excitability in cells that are suddenly changed from a heterozygous (wild-type) state to a homozygous mutant state, an estimate of the stability and mean lifetime of the calcium channel has been obtained. During rapid growth, channel dilution due to division occurred, but no channel decay was observed. Under conditions of slow growth, decay could also be observed; channel lifetime was found to be from 5 to 8 days.


Amyloid goiter and hypopituitarism in a patient with systemic amyloidosis.
Systemic amyloidosis may infiltrate the thyroid or other endocrine glands but rarely causes endocrine dysfunction. We describe a 45 years old female patient with diffusely enlarged goiter and hypopituitarism secondary to amyloid infiltration of the thyroid gland and possibly pituitary gland, respectively. She was on chronic haemodialysis for 3 years due to systemic amyloidosis. While she was being prepared for thyroidectomy, adrenal failure developed. Her anterior pituitary hormone levels were low and magnetic resonance imaging of the hypophysis showed low signal intensity in right part of the adenohypophysis. She improved with corticosteroid replacement therapy and underwent subtotal thyroidectomy without any complication. Histopathologically, amyloid deposition was demonstrated in the thyroid gland. To our knowledge, this is the first case with amyloid goiter and hypopituitarism secondary to systemic amyloidosis. Amyloid infiltration should be considered in a systemic amyloidosis patient presenting with rapidly enlarged thyroid gland and signs of hypopituitarism.


Deformation characteristics of solid-state benzene as a step towards understanding planetary geology.
Small organic molecules, like ethane and benzene, are ubiquitous in the atmosphere and surface of Saturn's largest moon Titan, forming plains, dunes, canyons, and other surface features. Understanding Titan's dynamic geology and designing future landing missions requires sufficient knowledge of the mechanical characteristics of these solid-state organic minerals, which is currently lacking. To understand the deformation and mechanical properties of a representative solid organic material at space-relevant temperatures, we freeze liquid micro-droplets of benzene to form ~10 μm-tall single-crystalline pyramids and uniaxially compress them in situ. These micromechanical experiments reveal contact pressures decaying from ~2 to ~0.5 GPa after ~1 μm-reduction in pyramid height. The deformation occurs via a series of stochastic (~5-30 nm) displacement bursts, corresponding to densification and stiffening of the compressed material during cyclic loading to progressively higher loads. Molecular dynamics simulations reveal predominantly plastic deformation and densified region formation by the re-orientation and interplanar shear of benzene rings, providing a two-step stiffening mechanism. This work demonstrates the feasibility of in-situ cryogenic nanomechanical characterization of solid organics as a pathway to gain insights into the geophysics of planetary bodies.


Pre-existing liver cirrhosis reduced the toxic effect of diethylene glycol in a rat model due to the impaired hepatic alcohol dehydrogenase.
Hepatic metabolizing enzymes of diethylene glycol (DEG) are impaired in liver diseases. Thus, the purpose of this study was to increase our understandings in metabolism and toxicology of DEG by clarifying the influences of pre-existing liver disease. Forty Sprague-Dawley rats with carbon tetrachloride-induced liver cirrhosis and 20 control rats were intraperitoneally administered a single dose of DEG, and randomly killed 1, 2, 5 or 8 days following exposure. Compared with control rats, the model rats had significantly higher blood CO(2)-combining power, lower blood urine nitrogen, serum creatinine and alanine aminotransferase levels on the second day and a lower mortality rate on the eighth day following DEG exposure. Enlargements of liver and kidneys and degeneration and necrosis of hepatocytes and renal tubules in the model rats was also less serious than in the control rats. Urine DEG levels were significantly higher on the first day in the model rats than the control rats (46.65 ± 8.79 mg vs 18.88 ± 6.18 mg, p < 0.01), but DEG concentrations in the blood, liver and kidneys were lower. Hepatic alcohol dehydrogenase (ADH) activity in the model rats was significantly lower than that in the control rats, which was positively related to renal damage. The toxic effects of DEG in rats with pre-existing liver cirrhosis are significantly reduced, which may be due to the decreased hepatic ADH activity. It suggests that the metabolite of ADH is responsible for DEG poisoning, and this toxic metabolite may mainly originate in the liver.


Epidemiology and impact of rheumatic disorders in the United States Hispanic population.
The emergence of a sizable Hispanic population in the US is a relatively recent historical phenomenon, and thus much is still unknown about this group of North Americans. Data from national surveys suggest small differences between Hispanic and non-Hispanic white populations in the age-adjusted prevalence of self-reported arthritic conditions. However, the rate of activity-limitation attributable to arthritis is higher among Hispanic patients. This likely reflects the poorer socioeconomic conditions and lack of health insurance that prevail among Hispanic populations, which may limit their access to rheumatologic care. Osteoporotic vertebral and hip fractures are less frequent, and proximal femoral mineral density is higher, in Hispanic individuals than in non-Hispanic white individuals. The mechanisms for these observations are currently under investigation. There have been no studies of the prevalence of osteoarthritis, rheumatoid arthritis, or systemic lupus erythematosus among Hispanic populations. However, important immunogenetic, clinical, and psychosocial differences between Hispanic and non-Hispanic patients in regard to rheumatoid arthritis and systemic lupus erythematosus have been reported. There is no published information on the prevalence or characteristics of other rheumatic diseases in the US Hispanic population. Emerging evidence suggests considerable underuse of certain health services for arthritis among Hispanic patients, likely due in part to socioeconomic factors. Further research is needed to determine whether biologic, cultural or psychosocial factors contribute to underuse as well. There is clearly a need for data on the prevalence and characteristics of arthritis and other rheumatic and musculoskeletal diseases in this emerging US population.


A computer-generated supercoiled model of the pUC19 plasmid.
DNA models have become a powerful tool in the simulation of radiation-induced molecular damage. Here, a computer code was developed which calculates the coordinates of individual atoms in supercoiled plasmid DNA. In this prototype study, the known base-pair sequence of the pUC19 plasmid has been utilized. The model was built in a three-step process. Firstly, a Monte Carlo simulation was performed to shape a segment chain skeleton. Checks on elastic energy, distance and unknotting were applied. The temperature was considered in two different ways: (1) it was kept constant at 293 K and (2) it was gradually reduced from 350 K to less than 10 K. Secondly, a special smoothing procedure was introduced here to remove the edges from the segment chain without changing the total curve length while avoiding the production of overshooting arcs. Finally, the base pair sequence was placed along the smoothed segment chain and the positions of all the atoms were calculated. As a first result, a few examples of the supercoiled plasmid models will be presented, demonstrating the strong influence of appropriate control of the system temperature.


Detection of Serum Antibodies to Hepatitis E Virus Based on HEV Genotype 3 ORF2 Capsid Protein Expressed in Nicotiana benthamiana.
Hepatitis E virus (HEV) causes epidemics in developing countries and is primarily transmitted through the fecal-oral route. There have been recent reports on the zoonotic spread of the virus, and several animal species, primarily pigs, have been recognized as reservoirs of HEV. Because of its possible spread, there is an urgent need of a method for the cost-effective production of HEV proteins that can be used as diagnostic antigens for the serological detection of anti-HEV antibodies. The HEV open reading frame (ORF)2 protein was purified from plant tissue by using immobilized metal-anion chromatography (IMAC). The recombinant protein was used to develop an in-house ELISA for testing anti-HEV antibodies in both human and swine sera. Thirty-six serum samples collected from patients with serologically proven HEV infection with commercial kits were tested for anti-HEV IgG antibodies by using the plant-expressed protein. Forty-five serum samples collected from apparently healthy pigs in Bulgarian farms were also tested. We confirmed the transient expression and purification of a truncated version of the HEV genotype 3 capsid protein in Nicotiana benthamiana and its usefulness as a diagnostic antigen. ELISA showed the presence of anti-HEV IgG antibodies in 29 of the 36 human samples. The in-house ELISA showed anti-HEV IgG antibodies in 34 of the 45 pigs. We describe a method for the production of HEV ORF2 protein in N. benthamiana and the usefulness of this protein for the serological detection of anti-HEV antibodies in both humans and swine.


Berson, Yalow, and the JCI: the agony and the ecstasy.
The isolation of insulin in 1921 by Banting, Best, Collip, and Macleod stands as one of the most dramatic stories in modern medical investigation. Only two years passed between the initial experiments in dogs to widespread human application to the awarding of the Nobel Prize in 1923. Insulin-related research has also served as a focus, at least in part, for the work of three other Nobel Prize recipients: determination of the chemical structure of insulin by Frederick Sanger in 1958; determination of the three-dimensional structures of insulin and vitamin B12 by Dorothy Hodgkin in 1964; and finally, the development of immunoassay by Solomon Berson and Rosalyn Yalow in 1959-1960, which led to a Nobel Prize for Yalow in 1977 (five years after the untimely death of Berson). The history of Yalow and Berson's discovery and its impact on the field is an illustration of the adage that every story has two sides.


The influence of habitats on female mobility in Central and Western Africa inferred from human mitochondrial variation.
When studying the genetic structure of human populations, the role of cultural factors may be difficult to ascertain due to a lack of formal models. Linguistic diversity is a typical example of such a situation. Patrilocality, on the other hand, can be integrated into a biological framework, allowing the formulation of explicit working hypotheses. The present study is based on the assumption that patrilocal traditions make the hypervariable region I of the mtDNA a valuable tool for the exploration of migratory dynamics, offering the opportunity to explore the relationships between genetic and linguistic diversity. We studied 85 Niger-Congo-speaking patrilocal populations that cover regions from Senegal to Central African Republic. A total of 4175 individuals were included in the study. By combining a multivariate analysis aimed at investigating the population genetic structure, with a Bayesian approach used to test models and extent of migration, we were able to detect a stepping-stone migration model as the best descriptor of gene flow across the region, with the main discontinuities corresponding to forested areas. Our analyses highlight an aspect of the influence of habitat variation on human genetic diversity that has yet to be understood. Rather than depending simply on geographic linear distances, patterns of female genetic variation vary substantially between savannah and rainforest environments. Our findings may be explained by the effects of recent gene flow constrained by environmental factors, which superimposes on a background shaped by pre-agricultural peopling.


Safety of Tubal Occlusion by Minilaparotomy Provided by Trained Clinical Officers Versus Assistant Medical Officers in Tanzania: A Randomized, Controlled, Noninferiority Trial.
Tubal occlusion by minilaparotomy is a safe, highly effective, and permanent way to limit childbearing. We aimed to establish whether the safety of the procedure provided by trained clinical officers (COs) was not inferior to the safety when provided by trained assistant medical officers (AMOs), as measured by major adverse event (AE) rates. In this randomized, controlled, open-label noninferiority trial, we enrolled participants at 7 health facilities in Arusha region, Tanzania, as well as during outreach activities conducted in Arusha and neighboring regions. Consenting, eligible participants were randomly allocated by a research assistant at each site to minilaparotomy performed by a trained CO or by a trained AMO, in a 1:1 ratio. We asked participants to return at 3, 7, and 42 days postsurgery. The primary outcome was the rate of major AEs following minilaparotomy performed by COs versus AMOs, during the procedure and through 42 days follow-up. The noninferiority margin was 2%. The trial is registered with ClinicalTrials.gov, Identifier NCT02944149. We randomly allocated 1,970 participants between December 2016 and June 2017, 984 to the CO group and 986 to the AMO group. Most (87%) minilaparotomies were conducted during outreach services. In the intent-to-treat analysis, 0 of 978 participants had a major AE in the CO group compared with 1 (0.1%) of 984 in the AMO group (risk difference: -0.1% [95% confidence interval: -0.3% to 0.1%]), meeting the criteria for noninferiority. We saw no evidence of differences in measures of procedure performance, participant satisfaction, or provider self-efficacy between the groups. Tubal occlusion by minilaparotomy performed by trained COs is safe, effective, and acceptable to women, and the procedure can be safely and effectively provided in outreach settings. Our results provide evidence to support policy change in resource-limited settings to allow task shifting of minilaparotomy to properly trained and supported COs, increasing access to female sterilization and helping to meet the rising demand for the procedure among women wanting to avoid pregnancy. They also suggest high demand for these services in Tanzania, given the large number of women recruited in a relatively short time period.


Biodemographic trajectories of age-specific reproliferation from stationary phase in the yeast Saccharomyces cerevisiae seem multiphasic.
Ageing is usually seen as a monotonic decline of functions and survival. However, recent studies reported that age-specific mortality rates increased and then leveled off or even declined at later ages in several species including humans. Preliminary data using the yeast, Saccharomyces cerevisiae, demonstrated an even more complicated, non-monotonic pattern of reproliferation after stationary phase (i.e. the ability of a cell to exit stationary phase and form a colony). In the present article, we conducted a study of the age-specific reproliferation rates of yeast populations. Stationary phase yeast cells were maintained in water and the reproliferation rates were estimated by the number of yeast able to exit stationary phase on rich growth media. We showed that the age-specific reproliferation rates in yeast seem to rise, fall and rise again. Furthermore, we observed this pattern in different experiments and in different genotypes and established that this pattern was not due to genetic heterogeneity of the populations.


Effect of dietary crude protein modification on ammonia and nitrous oxide concentration on a tie-stall dairy barn floor.
Dietary crude protein (CP) reduction is considered a useful strategy to minimize cow N excretion and NH(3) and N(2)O emissions. The aim of the current work was to relate dietary CP modification to whole-animal N balance and subsequent NH(3) and N(2)O concentrations on a tie-stall barn floor. The effect of temperature on NH(3) and N(2)O concentration was also studied. Three Holstein mid to late lactating cows were confined in separate tie-stalls and randomly assigned to 3 diets with varying CP content [low CP (LCP): 14.1%; moderate CP (MCP): 15.9%; high CP (HCP): 16.9%]. Increasing N intake (from 438.6 to 522.8 g of N/d) improved milk yield (from 22.1 to 24.2 kg/d). However, N use efficiency tended to decrease with increasing dietary CP, as shown by milk N use efficiency (from 23.9 to 22.6%), milk urea N (from 15.4 to 18.7 mg/dL), and excreted N per milk yield unit (from 14.7 to 16.4 g of N/kg of milk). Because of higher N excretion, NH(3) concentration on the dairy barn floor increased (LCP: 7.1mg of NH(3)/m(3); MCP: 10.4 mg of NH(3)/m(3); HCP: 10.8 mg of NH(3)/m(3)). In contrast, N(2)O concentration did not respond to dietary manipulation (mean 1.1mg of N(2)O/m(3)). Temperature, which ranged between 12.6 and 18.0 degrees C, did not affect NH(3) and N(2)O concentrations at the stall level. However, when fecal and urinary samples were incubated at 4, 19, and 29 degrees C in the laboratory, ammonia concentration increased for all diets, especially for the MCP and HCP diets, as the temperature increased. In contrast, N2O concentration was negatively related to increasing temperature. In conclusion, data from the current trial demonstrate that lowering dietary CP minimizes NH(3) concentration on dairy stall floors although temperature controls the rate of NH(3) volatilization. On the other hand, N(2)O concentration is not affected by dietary treatments on tie-stall floors.


Retention characteristics of protonated mobile phases injected into deuterated mobile phases in capillary liquid chromatography (LC) using on-line nuclear magnetic resonance (NMR) detection.
The behavior of protonated binary solvents injected into deuterated binary mobile phases in capillary LC is studied with NMR. Specifically, the solvent elution is followed on-flow with a capillary LC coupled to a 900 nL volume microcoil NMR probe. A range of identical composition 5% protonated (and 95% deuterated) solvents is injected into composition-matched deuterated mobile phases of CD(3)CN/D(2)O and CD(3)OD/D(2)O. The protonated components separate for all solvent combinations except at 80% CD(3)CN/20% D(2)O and similar to 72% CD(3)OD/28% D(2)O where only a single retention time is observed. The more hydrophilic protonated component, HOD, elutes first with higher percentages of hydrophilic solvent, D(2)O, in the mobile phase whereas retention is reversed with the higher percentage of the more hydrophobic solvent (CD(3)CN and CD(3)OD) in the mobile phase. The hydrophilic/hydrophobic nature of the chromatographic system as a function of mobile phase composition is characterized by following the retention times of protonated solvents.


Debate rages over pharmaceutical coverage for seniors.
Seeking to make good on his promise to provide an interim pharmaceutical benefit for America's seniors, President George W. Bush recently outlined a new prescription drug proposal utilizing private pharmacy benefit managers (PBM). This new plan is designed to make prescription drugs more affordable by setting up a government-endorsed discount program. Here's an overview of the plan and a look to the future debate.


Activation of mouse and human peroxisome proliferator-activated receptors (PPARs) by phthalate monoesters.
Administration of phthalates is known to cause toxicity and liver cancer in rodents through the activation of peroxisome proliferator-activated receptors (PPARs), and the monoesters appear to be the active metabolites that function as ligands of PPARs. There is evidence that PPARs exhibit significant species differences in response to ligand activation. In this study, the activation of mouse and human PPARalpha, PPARbeta, and PPARgamma by a broad class of phthalate monoesters was investigated using a trans-activation assay, functional analysis of PPARalpha target gene expression, and a PPARgamma-mediated differentiation assay. These studies demonstrated a range in the ability of various phthalate monoesters to activate PPARalpha, with the mouse PPARalpha generally being activated at lower concentrations and exhibiting a greater response than human PPARalpha. Similarly, a range in the trans-activation of mouse PPARbeta by phthalate monoesters was also observed, but this effect was not found with human PPARbeta. A number of phthalate monoesters activated both mouse and human PPARgamma, with similar sensitivity being exhibited by both receptors. These studies show that the potency and efficacy of phthalate monoesters for the activation of PPARalpha and PPARgamma increase with increasing side-chain length. These studies also show that mouse PPARalpha and PPARbeta are generally activated at lower concentrations of phthalate monoesters than human PPARalpha and PPARbeta, and that both mouse and human PPARgamma exhibit similar sensitivity to phthalate monoesters. Lastly, there is a good relationship between the relative ability of phthalate monoesters to trans-activate PPARalpha and PPARgamma, and the relative induction of PPARalpha target gene mRNA and PPARgamma-mediated adipocyte differentiation, respectively.


Patterns of kinase activation induced by injury in the murine femoral artery.
Intimal hyperplasia remains the principal lesion in the development of restenosis after vessel wall injury. Cell signaling in vascular smooth muscle cells remains a potential molecular target to modulate the development of intimal hyperplasia. The aim of this study was to define a baseline pattern of histological changes and kinase activation in a murine model. The murine femoral wire injury model was used in which a microwire was passed through a branch of the femoral artery and used to denude the common femoral artery. Pluronic gel was used to apply mitogen-activated protein kinases (MAPK) inhibitors (PD98059, SB230580, and SP600125) on the exterior of the vessels. Specimens were perfusion-fixed and sections were stained for morphometry using an ImagePro system. Additional specimens of femoral artery were also harvested and snap frozen for Western blotting and zymography to allow for the study of kinase and protease activation. Contralateral vessels were used as controls. The injured femoral arteries developed intimal hyperplasia, which is maximal at 28 days and does not change substantially between day 28 and day 56. Sham-operated vessels did not produce such a response. Cell apoptosis peaked within 3 days and cell proliferation peaked at 7 days after injury. There is a time-dependent increase in kinase activity immediately after injury. MEK1/2 activation peaks at 20 min after injury and is followed by a peak in extracellular signal-regulated kinase-1/2 activation at 45 min. The stress kinases p38(MAPK) and JNK peak between 10 and 20 min. Activation of akt is later at 45 min and 120 min and activation of p70S6K was biphasic. There was a time-dependent increase in uPA/PAI-1 expression and activity after injury. Local application of MAPK inhibitors (PD98059, SB230580, and SP600125) within a pluronic gel reduced respective MAPK activity, decreased cell proliferation and enhanced cell apoptosis, increased PAI-1, and decreased uPA expression and activity; at 14 days there was a decrease in intimal hyperplasia. These data demonstrate that femoral wire injury in the mouse induces a consistent model of intimal hyperplasia and that it is associated with a time dependent increase in signaling kinase activity. Interruption of these pathways will interrupt the uPA/PAI-1 pathway and decrease intimal hyperplasia development. Accurate characterization of cell signaling is a necessary step in the development of molecular therapeutics.


Long-range steady-state density profiles induced by localized drive.
We show that the presence of a localized drive in an otherwise diffusive system results in steady-state density and current profiles that decay algebraically to their global average value, away from the drive in two or higher dimensions. An analogy to an electrostatic problem is established, whereby the density profile induced by a driving bond maps onto the electrostatic potential due to an electric dipole located along the bond. The dipole strength is proportional to the drive, and is determined self-consistently by solving the electrostatic problem. The profile resulting from a localized configuration of more than one driving bond can be straightforwardly determined by the superposition principle of electrostatics. This picture is shown to hold even in the presence of exclusion interaction between particles.


Safety and Effectiveness of the Denali Inferior Vena Cava Filter: Intermediate Follow-Up Results.
The purpose of the study is to evaluate the clinical safety and effectiveness of the Denali (Bard, Tempe, Arizona) retrievable inferior vena cava (IVC) filter. In this retrospective study, authors reviewed the data of Denali IVC filters placed at their institution between 2013 and 2015. The clinical presentation, indications, and procedure-related complications during placement and retrieval were evaluated. The frequency of post filter pulmonary embolism (PE) and filter-related complications was assessed. Denali filters were placed in 87 patients (47 males; mean age: 56 years). Twenty patients presented with PE, 45 with deep vein thrombosis (DVT), and 21 with both PE and DVT, 1 filter was placed prophylactically before surgery. Indications for filter placement included contraindications to anticoagulation (AC; n = 80), failure of AC (n = 4), and complications of AC (n = 3). No patients had PE on follow-up imaging after filter placement. Retrieval was attempted in 31 patients after a mean period of 125 days (range: 34-324 days). The filter was successfully removed in 31 (100%) patients. Follow-up imaging, available in 71 (82%) patients (range: 2-538 days), demonstrated penetration of 15 legs in 5 patients, caval thrombus in 3, 1 resulting in caval occlusion, <15° filter tilt in 5, and no leg fractures or crossed legs. The Denali filter is safe during deployment and readily retrievable. The overall safety following deployment is similar to those reported in the literature, and the incidence of filter fractures and migration appears to be less than the previous generation of Bard devices.