Historical perspectives on the roots of the apical negative pressure irrigation technique in endodontics.
The removal of the infectious process caused by an abscess in the periapical tissues was a challenge to dentists in the early part of the 20th century. While they recognized the need to debride the periapical tissues, the process was slow, tedious, and often fraught with failure that resulted in tooth extraction. However, with some creative ingenuity, an irrigation-suction apparatus was developed in the 1930s that enabled rapid and thorough debridement. This appliance went through multiple developmental permutations and was successful in achieving the desired goal. Interestingly, while the purpose of this device was a controlled periapical debridement through the root canal, and not necessarily a focus on a cleaning of the intricacies of the root canal system, the basic concept purported was similar to contemporary approaches used in root canal debridement today.


Double-image encryption without information disclosure using phase-truncation Fourier transforms and a random amplitude mask.
We present a study about information disclosure in phase-truncation-based cryptosystems. The main information of the original image to be encoded can be obtained by using a decryption key in the worst case. The problem cannot be thoroughly solved by imaginary part truncating, keeping the encryption keys as private keys, or applying different phase keys for different plaintexts during each encryption process as well as the phase modulation in the frequency domain. In order to eliminate the risk of unintended information disclosure, we further propose a nonlinear spatial and spectral encoding technique using a random amplitude mask (RAM). The encryption process involving two security layers can be fully controlled by a RAM. The spatial encoding of the plaintext images and the simultaneous encryption of the plaintext images and the encryption key greatly enhance the security of system, avoiding several attacks that have cracked the phase-truncation-based cryptosystems. Besides, the hybrid encryption system retains the advantage of a trap door one-way function of phase truncation. Numerical results have demonstrated the feasibility and effectiveness of the proposed encryption algorithm.


Regulation of estrogen receptor concentration and activity by an erbB/HER ligand in breast carcinoma cell lines.
Expression of the erbB-2 oncogene in breast cancer patients correlates with poor prognosis and failure of hormonal therapy. In this study, the effects of a putative erbB/HER ligand, gp30, on estrogen receptor (ER) concentration and activity was investigated in the estrogen receptor positive human breast cancer cells, BT474 and MCF-7, which express either high or low levels of erbB-2 and erbB-4, respectively. Treatment of cells with gp30 resulted in a decrease in the steady-state level of estrogen receptor protein by approximately 70-80%. The effect of gp30 on the concentration of ER was independent of serum in the media and was not inhibited by an epidermal growth factor receptor blocking antibody. In addition to the effect on ER protein, gp30 decreased the steady-state level of ER messenger RNA. Transcription run on experiments demonstrated that the decrease in ER expression was mediated by a decrease in ER gene transcription. The effect of gp30 on estrogen receptor activity was also investigated in this study. Treatment of cells with gp30 blocked estradiol induction of progesterone receptor. Inhibition was observed at the level of progesterone receptor protein, messenger RNA, and gene transcription. gp30 also blocked estradiol induction of pS2 gene transcription. In addition to its effects on progesterone receptor and pS2, gp30 blocked activation of an estrogen response element in a transient transfection assay and inhibited ER binding to its response element in a DNA mobility shift assay, suggesting a direct effect on the estrogen receptor. The effects of gp30 on estrogen receptor concentration and activity were independent of the level of erbB-2 and erbB-4 in the cell. These data show that gp30 regulates the concentration of ER and modulates ER activity.


Plate Fixation of Expansive Open-Door Laminoplasty Decreases the Incidence of Postoperative C5 Palsy.
Retrospective study. To compare outcomes for plate fixation versus suture suspension in expansive open-door laminoplasty (ELAP) for cervical spondylotic myelopathy (CSM). C5 palsy and laminar closure have been reported to occur after ELAP when suture suspension is used to anchor the opened lamina. We assessed 174 patients with CSM who were treated by ELAP using either suture suspension (114 patients, mean age 63.3 y, 83 men and 31 women, mean follow-up 12.0 mo) or laminoplasty plate (60 patients, mean age 65.6 y, 45 men and 15 women, mean follow-up 12.6 mo) to maintain the position of the opened laminae. We compared clinical outcomes, radiographic findings, and complications between the 2 groups. Mean Japanese Orthopaedic Association scores had improved significantly for both groups at the final follow-up (P<0.001), with similar recovery rates: scores improved from 10.8±2.8 to 13.4±2.8 points in the suture-suspension (SS) group and from 11.3±2.7 to 13.6±2.2 points in the laminoplasty plate (LP) group. The incidence of C5 palsy was significantly higher in the SS group (7.9%) than in the LP group (1.7%; P<0.05). Computed tomography (CT) showed larger mean angles of the opened laminae in the SS group compared with the LP group (P<0.01). Magnetic resonance imaging (MRI) showed that the mean anterior spinal-cord space at the C5 level was larger in the SS group than in the LP group (P<0.01). Plate fixation in ELAP produced comparable clinical outcomes and significantly lowered the incidence of C5 palsy compared with suture suspension. CT and MRI findings indicated that laminoplasty plate fixation prevented excessive opening of the lamina and decreased dorsal spinal-cord shift, which might explain the lower incidence of C5 palsy.


CT-Based Assessment of Relative Soft-Tissue Alteration in Different Types of Ancient Mummies.
Mummification leads to alteration of soft-tissue morphology. No research has focused specifically on differences in soft-tissue shrinkage depending on mummification type. This study evaluated whether soft-tissue alteration is dependent on type of mummification. A total of 17 human mummies have been investigated by computed tomography (CT). Samples included artificially embalmed ancient Egyptian mummies, naturally mummified South American corpses, ice mummies (including the Iceman, South Tyrol Museum of Archeology, Bolzano, Italy, ca. 3,300 BC), bog bodies and a desiccated mummy of possibly Asian provenance. The acquired data were compared to four contemporary bodies. The extent of soft-tissue shrinkage was evaluated using CT data. Shrinkage was defined as soft-tissue relative to area of bone (in number of voxels). Measurements were taken at 13 anatomically defined locations. Ice mummies show the highest degree of preservation. This finding is most likely explained due to frozen water within tissues. All other types of mummies show significantly (at P < 0.05) smaller relative area of preserved soft-tissue. Variation between different anatomical structures (e.g., upper lip vs. mid-femur) is significant, unlike variation within one compartment (e.g., proximal vs. distal humerus). Mummification type strongly affects the degree of soft-tissue alteration, surprisingly mostly independent of overall historical age. These results highlight the unique morphological impact of taphonomy on soft-tissue preservation and are of particular interest in tissue research as well as in forensics.


The role of population movement in the epidemiology and control of schistosomiasis in Brazil: a preliminary typology of population movement.
This paper examines recent developments in migration studies. It reviews literature related to the potential role of internal population movement in the occurrence of schistosomiasis in Brazil and modifies Prothero's typology of population movement for use in Brazil. This modified classification system may contribute to a better understanding of schistosome transmission as well as improved research and control programs. The results of this study indicate that population movement in Brazil primarily involves economically-motivated rural-urban and interregional movement. However, several movement patterns have become increasingly important in recent years as a result of changing socioeconomic and urbanisation dynamics. These patterns include urban-urban, intracity and urban-rural movement as well as the movement of environmental refugees and tourists. Little is known about the epidemiological significance of these patterns. This paper also highlights the role of social networks in the decision to migrate and to settle. Prothero's classic population movement typology categorises movement as either one-way migrations or circulations and examines them along spatial and temporal scales. However, the typology must be modified as epidemiological information about new patterns becomes available. This paper identifies areas that require further research and offers recommendations that can improve the measurement and spatial analysis of the relationship between population movement and schistosomiasis.


Buprenorphine: clinical pharmacokinetics in the treatment of opioid dependence.
Buprenorphine is a semi-synthetic opioid derived from thebaine, a naturally occurring alkaloid of the opium poppy, Papaver somniferum. The pharmacology of buprenorphine is unique in that it is a partial agonist at the opioid mu receptor. Buprenorphine undergoes extensive first-pass metabolism and therefore has very low oral bioavailability; however, its bioavailability sublingually is extensive enough to make this a feasible route of administration for the treatment of opioid dependence. The mean time to maximum plasma concentration following sublingual administration is variable, ranging from 40 minutes to 3.5 hours. Buprenorphine has a large volume of distribution and is highly protein bound (96%). It is extensively metabolised by N-dealkylation to norbuprenorphine primarily through cytochrome P450 (CYP) 3A4. The terminal elimination half-life of buprenorphine is long and there is considerable variation in reported values (mean values ranging from 3 to 44 hours). Most of a dose of buprenorphine is eliminated in the faeces, with approximately 10-30% excreted in urine. Naloxone has been added to a sublingual formulation of buprenorphine to reduce the abuse liability of the product. The presence of naloxone does not appear to influence the pharmacokinetics of buprenorphine. Buprenorphine crosses the placenta during pregnancy and also crosses into breast milk. Buprenorphine dosage does not need to be significantly adjusted in patients with renal impairment; however, since CYP3A activity may be decreased in patients with severe chronic liver disease, it is possible that the metabolism of buprenorphine will be altered in these patients. Although there is limited evidence in the literature to date, drugs that are known to inhibit or induce CYP3A4 have the potential to diminish or enhance buprenorphine N-dealkylation. It appears that the interaction between buprenorphine and benzodiazepines is more likely to be a pharmacodynamic (additive or synergistic) than a pharmacokinetic interaction. The relationship between buprenorphine plasma concentration and response in the treatment of opioid dependence has not been well studied. The pharmacokinetic and pharmacodynamic properties of buprenorphine allow it to be a feasible option for substitution therapy in the treatment of opioid dependence.


Observing of the asymmetry of hippocampal formation on patients with complex partial attacks.
Lobus limbicus is anatomical basis for explaining the temporal epilepsy because it not only includes the focus of infection of temporal lobe but also the frontal lobe, and with it we can explain many of the phenomenon's of the epilepsy (hallucinations, the change of the effects, and so on.). The goal of this assignment was to explore the asymmetry of hippocampal formation on the patients with complex partial attacks. The results show that the least number of patients with epilepsy have a symmetric (same) size of the hippocampal formation both from the left and the right side. The number is statistically significantly lower than the number of patients with epilepsy who has asymmetric (different size) hippocampal formation both from the left and the right side. By the direction of asymmetry the difference in the number of patients with epilepsy isn't statistically significant. Coefficient. of asymmetry shows that the asymmetry on the left side is more common to men, while it is distributed evenly on sides, left and right in women. Testing of significance in age differences depends on the sim./asim. of the hippocampal formation of both the right and the left side of coronal slice is a statistical significance. While with axial and sagittal slices, we have a statistical significance between a women and a man on the level of the course of asymmetry. So, the use of MRI technique in examining asymmetry of the hippocampal formation, that we used on this asymmetry, we suggest it as an template of future examinations in a sense of shedding light of the anatomical functions that is located on the basis of neuropsychiatrics dysfunctions.


Dynamic stabilization in 1sigma(u)-->1pi(g) excited nitrogen clusters.
High-resolution 1s near-edge spectra of molecular nitrogen and variable size nitrogen clusters obtained using monochromatic synchrotron radiation from the high brilliance BESSY-II storage ring facility are reported. The vibrationally resolved 1sigma(u)-->1pi(g) core-to-valence excitation band of clusters shows a distinct redshift of 6+/-1 meV relative to the isolated molecule, but the vibrational structure and linewidths are essentially unchanged. This shift is assigned to dynamic stabilization of 1sigma(u)-->1pi(g) excited molecules in clusters, arising from the dynamic dipole moment generated by core-hole localization in the low-symmetry cluster field. This leads to changes in intermolecular interactions compared to the ground-state cluster. Such spectral shifts are expected to occur generally in molecular clusters and in the corresponding condensed phase.


Effect of hyperoxia and hypercapnia on tissue oxygen and perfusion response in the normal liver and kidney.
Inhalation of air with altered levels of oxygen and carbon dioxide to manipulate tissue oxygenation and perfusion has both therapeutic and diagnostic value. These physiological responses can be measured non-invasively with magnetic resonance (MR) relaxation times. However, interpreting MR measurements is not straight-forward in extra-cranial organs where gas challenge studies have only begun to emerge. Inconsistent results have been reported on MR, likely because different organs respond differently. The objective of this study was to elucidate organ-specific physiological responses to gas challenge underlying MR measurements by investigating oxygenation and perfusion changes in the normal liver and kidney cortex. Gas challenges (100% O(2), 10% CO(2), and carbogen [90% O(2)+10% CO(2)]) interleaved with room air was delivered to rabbits to investigate their effect on tissue oxygenation and perfusion. Real-time fiber-optic measurements of absolute oxygen and relative blood flow were made in the liver and kidney cortex. Only the liver demonstrated a vasodilatory response to CO(2). Perfusion changes to other gases were minimal in both organs. Tissue oxygenation measurements showed the liver responding only when CO(2) was present and the kidney only when O(2) was present. This study reveals distinct physiological response mechanisms to gas challenge in the liver and kidney. The detailed characterization of organ-specific responses is critical to improving our understanding and interpretation of MR measurements in various body organs, and will help broaden the application of MR for non-invasive studies of gas challenges.


Retinoic acid-inducible G protein-coupled receptors bind to frizzled receptors and may activate non-canonical Wnt signaling.
Frizzled (Fz) seven-pass transmembrane receptors are Wnt receptors and function in a variety of developmental pathways. Here we identify retinoic acid-inducible gene-1, 2, 3, and 4 (RAIG1, 2, 3, and 4) as potential Fz binding proteins. RAIG proteins are seven-pass transmembrane receptors, and Xenopus RAIG2, 3, and 4 are expressed in early gastrula. XRAIG2 can activate small GTPases, such as RhoA, Rac, and Cdc42, and c-jun N-terminal kinase, thus exhibit activities that overlap with non-canonical Wnt/Fz signaling. Injection of XRAIG2 mRNA into Xenopus embryo causes a severe shortened and bent body axis due to defective gastrulation movements, reminiscent of abnormal non-canonical Wnt signaling. XRAIG2 affects convergent extension in activin-treated animal caps, which can be partially rescued by co-injection of a dominant-negative form of Cdc42. In zebrafish embryo, XRAIG2 also causes Ca(2+) flux, one of the consequences of non-canonical Wnt signaling. These results suggest a possible crosstalk/integration between Wnt/Frizzled and RAIG signal transduction pathways.


Simple Tip-Based Sample Processing Method for Urinary Proteomic Analysis.
Mass spectrometry-based urinary proteomics is one of the most attractive strategies to discover proteins for diagnosis, prognosis, monitoring, or prediction of therapeutic responses of urological diseases involving the kidney, prostate, and bladder; however, interfering compounds found in urine necessitate sample preparation strategies that are currently not suitable for urinary proteomics in the clinical setting. Herein, we describe the C4-tip method, comprising a simple, automated strategy utilizing a reverse-phase resin tip-based format and "on-tip" digestion to examine the urine proteome. We first determined the optimal conditions for protein isolation and protease digestion on the C4-tip using the standard protein bovine fetuin. Next, we applied the C4-tip method to urinary proteomics, identifying a total of 813 protein groups using LC-MS/MS, with identified proteins from the C4-tip method displaying a similar distribution of gene ontology (GO) cellular component assignments compared to identified proteins from an ultrafiltration preparation method. Finally, we assessed the reproducibility of the C4-tip method, revealing a high Spearman correlation R-value for shared proteins identified across all tips. Together, we have shown the C4-tip method to be a simple, robust method for high-throughput analysis of the urinary proteome by mass spectrometry in the clinical setting.


Text-Messaging, Online Peer Support Group, and Coaching Strategies to Optimize the HIV Prevention Continuum for Youth: Protocol for a Randomized Controlled Trial.
America's increasing HIV epidemic among youth suggests the need to identify novel strategies to leverage services and settings where youth at high risk (YAHR) for HIV can be engaged in prevention. Scalable, efficacious, and cost-effective strategies are needed, which support youth during developmental transitions when risks arise. Evidence-based behavioral interventions (EBIs) have typically relied on time-limited, scripted, and manualized protocols that were often delivered with low fidelity and lacked evidence for effectiveness. This study aims to examine efficacy, implementation, and cost-effectiveness of easily mountable and adaptable, technology-based behavioral interventions in the context of an enhanced standard of care and study assessments that implement the guidelines of Centers for Disease Control and Prevention (CDC) for routine, repeat HIV, and sexually transmitted infection (STI) testing for high-risk youth. Youth aged between 12 and 24 years (n=1500) are being recruited from community-based organizations and clinics serving gay, bisexual, and transgender youth, homeless youth, and postincarcerated youth, with eligibility algorithms weighting African American and Latino youth to reflect disparities in HIV incidence. At baseline and 4-month intervals over 24 months (12 months for lower-risk youth), interviewers monitor uptake of HIV prevention continuum steps (linkage to health care, use of pre- or postexposure prophylaxis, condoms, and prevention services) and secondary outcomes of substance use, mental health, and housing security. Assessments include rapid diagnostic tests for HIV, STIs, drugs, and alcohol. The study is powered to detect modest intervention effects among gay or bisexual male and transgender youth with 70% retention. Youth are randomized to 4 conditions: (1) enhanced standard of care of automated text-messaging and monitoring (AMM) and repeat HIV/STI testing assessment procedures (n=690); (2) online group peer support via private social media plus AMM (n=270); (3) coaching that is strengths-based, youth-centered, unscripted, based on common practice elements of EBI, available over 24 months, and delivered by near-peer paraprofessionals via text, phone, and in-person, plus AMM (n=270); and (4) online group peer support plus coaching and AMM (n=270). The project was funded in September 2016 and enrollment began in May 2017. Enrollment will be completed between June and August 2019. Data analysis is currently underway, and the first results are expected to be submitted for publication in 2019. This hybrid implementation-effectiveness study examines alternative models for implementing the CDC guidelines for routine HIV/STI testing for YAHR of acquiring HIV and for delivering evidence-based behavioral intervention content in modular elements instead of scripted manuals and available over 24 months of follow-up, while also monitoring implementation, costs, and effectiveness. The greatest impacts are expected for coaching, whereas online group peer support is expected to have lower impact but may be more cost-effective. ClinicalTrials.gov NCT03134833; https://clinicaltrials.gov/ct2/show/NCT03134833 (Archived by WebCite at http://www.webcitation.org/76el0Viw9). DERR1-10.2196/11165.


The occupational contribution to severe exacerbation of asthma.
The goal of this study was to identify occupational risk factors for severe exacerbation of asthma and estimate the extent to which occupation contributes to these events. The 966 participants were working adults with current asthma who participated in the follow-up phase of the European Community Respiratory Health Survey. Severe exacerbation of asthma was defined as self-reported unplanned care for asthma in the past 12 months. Occupations held in the same period were combined with a general population job-exposure matrix to assess occupational exposures. 74 participants reported having had at least one severe exacerbation event, for a 1-yr cumulative incidence of 7.7%. From regression models that controlled for confounders, the relative risk (RR) was statistically significant for low (RR 1.7, 95% CI 1.1-2.6) and high (RR 3.6, 95% CI 2.2-5.8) biological dust exposure, high mineral dust exposure (RR 1.8, 95% CI 1.02-3.2), and high gas and fumes exposure (RR 2.5, 95% CI 1.2-5.5). The summary category of high dust, gas, or fumes exposure had RR 3.1 (95% CI 1.9-5.1). Based on this RR, the population attributable risk was 14.7% among workers with current asthma. These results suggest occupation contributes to approximately one in seven cases of severe exacerbation of asthma in a working population, and various agents play a role.


Photosensitizer (PS)/polyhedral oligomeric silsesquioxane (POSS)-crosslinked nanohybrids for enhanced imaging-guided photodynamic cancer therapy.
Photodynamic therapy (PDT) has drawn extensive attention as a promising cancer treatment modality. However, most PDT nanoagents suffer from insufficient drug loading capacity, a severe self-quenching effect, premature release of drugs and/or potential toxicity. Herein, we rationally designed an inorganic-organic nanohybrid with high drug loading capacity and superior chemical stability for enhanced PDT. Polyhedral oligomeric silsesquioxane (POSS), an amine-containing cage-shaped building block, was crosslinked with chlorin e6 (Ce6), a carboxyl-containing photosensitizer, via the amine-carboxyl reaction. Polyethylene glycol (PEG) polymers were further modified on the surface of the nanoparticle to improve the aqueous dispersibility and prolong the circulation time of the final nanoconstruct (POSS-Ce6-PEG). The as-prepared POSS-Ce6-PEG has a considerably high loading rate of Ce6 (19.8 wt%) with desirable fluorescence emission and singlet oxygen generation. Besides, in vitro experiments revealed that the nanoagent exhibited enhanced cellular uptake and a preferred intracellular accumulation within mitochondria and the endoplasmic reticulum, resulting in high anticancer efficiency under light irradiation. Furthermore, in vivo imaging-guided PDT was also successfully achieved, showing the effective tumor targeting and ablation ability of POSS-Ce6-PEG. More importantly, the nanoagent possesses negligible dark cytotoxicity and systemic side effects. Therefore, POSS-Ce6-PEG as an eligible PDT theranostic agent holds great potential in clinical applications.


Adoption and implementation of physical activity and dietary counseling by community health center providers and nurses.
Primary care providers are expected to provide lifestyle counseling, yet many barriers exist. Few studies report on adoption and implementation in routine practice. This study reports training, adoption, and implementation of an intervention to promote physical activity (PA) and dietary counseling in community health centers. Providers (n = 30) and nurses (n = 28) from 9 clinics were invited to participate. Adopters completed CD-ROM training in stage-matched, patient-centered counseling and goal setting. Encounters were audio recorded. A subsample was coded for fidelity. Fifty-seven percent of providers and nurses adopted the program. Provider counseling was seen in 66% and nurse goal setting in 58% of participant (N = 266) encounters, although audio recordings were lower. Duration of provider counseling and nurse goal setting was 4.9 ± 4.5 and 7.3 ± 3.8 minutes, respectively. Most PA (80%) and diet (94%) goals were stage-appropriate. Although most providers discussed at least 1 behavioral topic, some topics (eg, self-efficacy, social support) were rarely covered. A sizeable percentage of providers and nurses completed training, rated it favorably, and delivered lifestyle counseling, although with variable fidelity. With low implementation cost and limited office time required, this model has the potential to be disseminated to improve counseling rates in primary care.


TRIM27 negatively regulates NOD2 by ubiquitination and proteasomal degradation.
NOD2, the nucleotide-binding domain and leucine-rich repeat containing gene family (NLR) member 2 is involved in mediating antimicrobial responses. Dysfunctional NOD2 activity can lead to severe inflammatory disorders, but the regulation of NOD2 is still poorly understood. Recently, proteins of the tripartite motif (TRIM) protein family have emerged as regulators of innate immune responses by acting as E3 ubiquitin ligases. We identified TRIM27 as a new specific binding partner for NOD2. We show that NOD2 physically interacts with TRIM27 via the nucleotide-binding domain, and that NOD2 activation enhances this interaction. Dependent on functional TRIM27, ectopically expressed NOD2 is ubiquitinated with K48-linked ubiquitin chains followed by proteasomal degradation. Accordingly, TRIM27 affects NOD2-mediated pro-inflammatory responses. NOD2 mutations are linked to susceptibility to Crohn's disease. We found that TRIM27 expression is increased in Crohn's disease patients, underscoring a physiological role of TRIM27 in regulating NOD2 signaling. In HeLa cells, TRIM27 is partially localized in the nucleus. We revealed that ectopically expressed NOD2 can shuttle to the nucleus in a Walker A dependent manner, suggesting that NOD2 and TRIM27 might functionally cooperate in the nucleus.We conclude that TRIM27 negatively regulates NOD2-mediated signaling by degradation of NOD2 and suggest that TRIM27 could be a new target for therapeutic intervention in NOD2-associated diseases.


Tumour-specific proline vulnerability uncovered by differential ribosome codon reading.
Tumour growth and metabolic adaptation may restrict the availability of certain amino acids for protein synthesis. It has recently been shown that certain types of cancer cells depend on glycine, glutamine, leucine and serine metabolism to proliferate and survive. In addition, successful therapies using L-asparaginase-induced asparagine deprivation have been developed for acute lymphoblastic leukaemia. However, a tailored detection system for measuring restrictive amino acids in each tumour is currently not available. Here we harness ribosome profiling for sensing restrictive amino acids, and develop diricore, a procedure for differential ribosome measurements of codon reading. We first demonstrate the functionality and constraints of diricore using metabolic inhibitors and nutrient deprivation assays. Notably, treatment with L-asparaginase elicited both specific diricore signals at asparagine codons and high levels of asparagine synthetase (ASNS). We then applied diricore to kidney cancer and discover signals indicating restrictive proline. As for asparagine, this observation was linked to high levels of PYCR1, a key enzyme in proline production, suggesting a compensatory mechanism allowing tumour expansion. Indeed, PYCR1 is induced by shortage of proline precursors, and its suppression attenuated kidney cancer cell proliferation when proline was limiting. High PYCR1 is frequently observed in invasive breast carcinoma. In an in vivo model system of this tumour, we also uncover signals indicating restrictive proline. We further show that CRISPR-mediated knockout of PYCR1 impedes tumorigenic growth in this system. Thus, diricore has the potential to reveal unknown amino acid deficiencies, vulnerabilities that can be used to target key metabolic pathways for cancer treatment.


Treatment strategies and outcomes for rectal villous adenoma from a single-center experience.
To analyze a 13-year, single-surgeon experience with villous adenoma of the rectum with respect to procedure, complications, recurrence, and cancer incidence. Retrospective review of patient and tumor characteristics, procedure, recurrence, and complications. University hospital. Patients who underwent excision of rectal villous adenoma. Complication, recurrence, and malignancy rates. Thirty-six patients underwent 30 transanal and 10 transabdominal excisions. Mean age was 66 years (range, 41-86 years) and mean follow-up was 25 months (range, 0.5-132 months). Mean tumor size was 3.0 cm (range, 0.5-11 cm) and the mean distance of the tumor from the anal verge was 4.9 cm (range, 0-10 cm). Preoperatively, 18 (45%) lesions harbored low-grade dysplasia while 17 (43%) had high-grade dysplasia. Postoperative pathology was discordant in 50% of patients, including 5 of 40 lesions (13%) that were recategorized as invasive cancer. Tumor size did not correlate with malignancy. The complication rate was significantly lower in transanal compared with transabdominal excisions (3.6% vs 50%, P = .005). There were 4 (12.5%) benign recurrences, all after transanal excisions. Complete excision is warranted for rectal villous adenomas, as biopsies were accurate only 50% of the time, and 1 in 8 patients had unsuspected cancer found after excision. Transanal excision with negative margins is associated with low recurrence and complication rates and is the preferred approach, even with large lesions.


Risk sensitivity for amounts of and delay to rewards: adaptation for uncertainty or by-product of reward rate maximising?
Observations that humans and other species are sensitive to variability in the outcome of their choices has led to the widespread assumption that this sensitivity reflects adaptations to cope with risk (stochasticity of action consequences). We question this assumption in experiments with starlings. We show that choices between outcomes that are risky in both amount and delay to food are predictable from preferences in the absence of risk. We find that the overarching best predictor of an option's value is the average of the ratios of amount to delay across its (frequency weighted) outcomes, an expression known as "Expectation of the Ratios", or EoR. Most tests of risk sensitivity focus on the predicted impact of energetic state on preference for risk. We show instead that under controlled state conditions subjects are variance- and risk-neutral with respect to EoR, and this implies variance neutrality for amounts and variance-proneness for delays. The weak risk aversion for amounts often reported requires a small modification of EoR. EoR is consistent with associative learning: acquisition of value for initially neutral stimuli is roughly proportional to the magnitude of their consequences and inversely proportional to the interval between the stimulus and its consequence's onset. If, as is likely, the effect of amount on acquisition is sublinear, the result is a deviation from EoR towards risk aversion for amount. In 3 experiments, we first establish individual birds' preferences between pairs of fixed options that differ in both amount and delay (small-sooner vs. large-later), and then examine choices between stochastic mixtures that include these options. Experiment 1 uses a titration to establish certainty equivalents, while experiments 2 and 3 measure degree of preference between options with static parameters. The mixtures differ in the coefficient of variation of amount, delay, or both, but EoR is sufficient to predict all results, with no additional explanatory role for riskiness.


The Mechanisms of Yu Ping Feng San in Tracking the Cisplatin-Resistance by Regulating ATP-Binding Cassette Transporter and Glutathione S-Transferase in Lung Cancer Cells.
Cisplatin is one of the first line anti-cancer drugs prescribed for treatment of solid tumors; however, the chemotherapeutic drug resistance is still a major obstacle of cisplatin in treating cancers. Yu Ping Feng San (YPFS), a well-known ancient Chinese herbal combination formula consisting of Astragali Radix, Atractylodis Macrocephalae Rhizoma and Saposhnikoviae Radix, is prescribed as a herbal decoction to treat immune disorders in clinic. To understand the fast-onset action of YPFS as an anti-cancer drug to fight against the drug resistance of cisplatin, we provided detailed analyses of intracellular cisplatin accumulation, cell viability, and expressions and activities of ATP-binding cassette transporters and glutathione S-transferases (GSTs) in YPFS-treated lung cancer cell lines. In cultured A549 or its cisplatin-resistance A549/DDP cells, application of YPFS increased accumulation of intracellular cisplatin, resulting in lower cell viability. In parallel, the activities and expressions of ATP-binding cassette transporters and GSTs were down-regulated in the presence of YPFS. The expression of p65 subunit of NF-κB complex was reduced by treating the cultures with YPFS, leading to a high ratio of Bax/Bcl-2, i.e. increasing the rate of cell death. Prim-O-glucosylcimifugin, one of the abundant ingredients in YPFS, modulated the activity of GSTs, and then elevated cisplatin accumulation, resulting in increased cell apoptosis. The present result supports the notion of YPFS in reversing drug resistance of cisplatin in lung cancer cells by elevating of intracellular cisplatin, and the underlying mechanism may be down regulating the activities and expressions of ATP-binding cassette transporters and GSTs.


Haplotype-specific MAPT exon 3 expression regulated by common intronic polymorphisms associated with Parkinsonian disorders.
Genome wide association studies have identified microtubule associated protein tau (MAPT) H1 haplotype single nucleotide polymorphisms (SNPs) as leading common risk variants for Parkinson's disease, progressive supranuclear palsy and corticobasal degeneration. The MAPT risk variants fall within a large 1.8 Mb region of high linkage disequilibrium, making it difficult to discern the functionally important risk variants. Here, we leverage the strong haplotype-specific expression of MAPT exon 3 to investigate the functionality of SNPs that fall within this H1 haplotype region of linkage disequilibrium. In this study, we dissect the molecular mechanisms by which haplotype-specific SNPs confer allele-specific effects on the alternative splicing of MAPT exon 3. Firstly, we use haplotype-hybrid whole-locus genomic MAPT vectors studies to identify functional SNPs. Next, we characterise the RNA-protein interactions at two loci by mass spectrometry. Lastly, we knockdown candidate splice factors to determine their effect on MAPT exon 3 using a novel allele-specific qPCR assay. Using whole-locus genomic DNA expression vectors to express MAPT haplotype variants, we demonstrate that rs17651213 regulates exon 3 inclusion in a haplotype-specific manner. We further investigated the functionality of this region using RNA-electrophoretic mobility shift assays to show differential RNA-protein complex formation at the H1 and H2 sequence variants of SNP rs17651213 and rs1800547 and subsequently identified candidate trans-acting splicing factors interacting with these functional SNPs sequences by RNA-protein pull-down experiment and mass spectrometry. Finally, gene knockdown of candidate splice factors identified by mass spectrometry demonstrate a role for hnRNP F and hnRNP Q in the haplotype-specific regulation of exon 3 inclusion. We identified common splice factors hnRNP F and hnRNP Q regulating the haplotype-specific splicing of MAPT exon 3 through intronic variants rs1800547 and rs17651213. This work demonstrates an integrated approach to characterise the functionality of risk variants in large regions of linkage disequilibrium.


Recruitment for a guided self-help binge eating trial: potential lessons for implementing programs in everyday practice settings.
To explore effects of various recruitment strategies on randomized clinical trial (RCT)-entry characteristics for patients with eating disorders within an everyday health-plan practice setting. Randomly selected women, aged 25-50, in a Pacific Northwest HMO were invited to complete a self-report binge-eating screener for two treatment trials. We publicized the trials within the health plan to allow self-referral. Here, we report differences on eating-disorder status by mode and nature of recruitment (online, mail, self-referred) and assessment (comprehensive versus abbreviated) and on possible differences in enrollee characteristics between those recruited by strategy (self-referred versus study-outreach efforts). Few differences emerged among those recruited through outreach who responded by different modalities (internet versus mail), early-versus-late responders, and those enrolling under more comprehensive or abbreviated assessment. Self-referred were more likely to meet binge-eating thresholds and reported higher average BMI than those recruited by outreach and responding by mail; however, in most respects the groups were more similar than anticipated. Fewer than 1% of those initially contacted through outreach enrolled. Aggressive outreach and screening is likely not feasible for broader dissemination in everyday practice settings and recruits individuals with more similar demographic and clinical characteristics to those recruited through more abbreviated and realistic screening procedures than anticipated.


The MEMA kwa Vijana project: design of a community randomised trial of an innovative adolescent sexual health intervention in rural Tanzania.
Effective interventions to reduce the incidence of HIV, other sexually transmitted infections (STIs) and unwanted pregnancy among adolescents in sub-Saharan Africa are urgently needed. This paper describes the rationale and design of a randomised trial of the impact of an innovative sexual health intervention among adolescents in rural Mwanza Region, Tanzania. The MEMA kwa Vijana intervention comprises a teacher-led, peer-assisted sexual health education programme for students in the last 3 years of primary school, training and supervision of health workers in the provision of youth-friendly health services, peer condom promotion and distribution, and wider community activities. Detailed process evaluation was conducted and the impact of the intervention was evaluated through a community-randomised trial in which a cohort of 9645 adolescents was followed up for 3 years. Both process and impact evaluation used multiple assessment methods. Impact measures included incidence and prevalence of HIV and other STIs, pregnancy rates, knowledge and reported attitudes and sexual behaviour, as well as qualitative assessments. Results of the baseline survey of the cohort have been presented previously. The outcome of the trial will be reported separately. Behaviour change interventions among adolescents have been widely advocated, but there have been few rigorously designed trials of their effectiveness, particularly in developing countries, and measurement of sexual behaviour is particularly problematic in this age group. The MEMA kwa Vijana trial was undertaken to address these problems and to collect rigorous evidence on the effectiveness of an innovative intervention, designed to be implemented on a very large scale.


Microchip-based structure determination of low-molecular weight proteins using cryo-electron microscopy.
Interest in cryo-Electron Microscopy (EM) imaging has skyrocketed in recent years due to its pristine views of macromolecules and materials. As advances in instrumentation and computing algorithms spurred this progress, there is renewed focus to address specimen-related challenges. Here we contribute a microchip-based toolkit to perform complementary structural and biochemical analysis on low-molecular weight proteins. As a model system, we used the SARS-CoV-2 nucleocapsid (N) protein (48 kDa) due to its stability and important role in therapeutic development. Cryo-EM structures of the N protein monomer revealed a flexible N-terminal "top hat" motif and a helical-rich C-terminal domain. To complement our structural findings, we engineered microchip-based immunoprecipitation assays that led to the discovery of the first antibody binding site on the N protein. The data also facilitated molecular modeling of a variety of pandemic and common cold-related coronavirus proteins. Such insights may guide future pandemic-preparedness protocols through immuno-engineering strategies to mitigate viral outbreaks.


Characterization of the lipid and protein organization in HBsAg viral particles by steady-state and time-resolved fluorescence spectroscopy.
Hepatitis B surface antigen (HBsAg) particles, produced in the yeast Hansenula polymorpha, are 20 nm particles, composed of S surface viral proteins and host-derived lipids. Since the detailed structure of these particles is still missing, we further characterized them by fluorescence techniques. Fluorescence correlation spectroscopy indicated that the particles are mainly monomeric, with about 70 S proteins per particle. The S proteins were characterized through the intrinsic fluorescence of their thirteen Trp residues. Fluorescence quenching and time-resolved fluorescence experiments suggest the presence of both low emissive embedded Trp residues and more emissive Trp residues at the surface of the HBsAg particles. The low emission of the embedded Trp residues is consistent with their close proximity in alpha-helices. Furthermore, S proteins exhibit restricted movement, as expected from their tight association with lipids. The lipid organization of the particles was studied using viscosity-sensitive DPH-based probes and environment sensitive 3-hydroxyflavone probes, and compared to lipid vesicles and low density lipoproteins (LDLs), taken as models. Like LDLs, the HBsAg particles were found to be composed of an ordered rigid lipid interface, probably organized as a phospholipid monolayer, and a more hydrophobic and fluid inner core, likely composed of triglycerides and free fatty acids. However, the lipid core of HBsAg particles was substantially more polar than the LDL one, probably due to its larger content in proteins and its lower content in sterols. Based on our data, we propose a structural model for HBsAg particles where the S proteins deeply penetrate into the lipid core.


Factors affecting compliance with the measles vaccination schedule in a Brazilian city.
The success of vaccination campaigns depends on the degree of adherence to immunization initiatives and schedules. Risk factors associated with children's failure to receive the measles vaccine at the correct age were studied in the city of São Paulo, Brazil. Case-control and exploratory study, in the metropolitan area of São Paulo. The caregivers of 122 children were interviewed regarding their perceptions and understanding about the measles vaccination and the disease. The results showed that age, region of residence, marital status and education level were unrelated to taking measles vaccines adequately. Most individuals remembered being informed about the last annual vaccination campaign by television, but no communication channel was significantly associated with vaccination status. The answers to questions about knowledge of the disease or the vaccine, when analyzed alone, were not associated with taking measles vaccinations at the time indicated by health agencies. The results showed that, when parents felt sorry for their children who were going to receive shots, they delayed the vaccination. Most of the children did not take the measles vaccination on the exactly recommended date, but delayed or anticipated the shots. It is clear that there is no compliance with the government's recommended measles vaccination schedule (i.e. first dose at nine and second at 15 months of age, as recommended in 1999 and 2000). Feeling sorry for the children receiving shots can delay vaccination taking.


Effects of various pollutant mixtures on immune responses of the blue mussel (Mytilus edulis) collected at a salinity gradient in Danish coastal waters.
The Baltic Sea is a semi-enclosed sea with a steady salinity gradient (3‰-30‰). Organisms have adapted to such low salinities, but are suspected to be more susceptible to stress. Within the frame of the integrated environmental monitoring BONUS + project "BEAST" the applicability of immune responses of the blue mussel was investigated in Danish coastal waters. The sampling sites were characterised by a salinity range (11-19‰) and different mixtures of contaminants (metals, PAHs and POPs), according to chemical analysis of mussel tissues. Variation partitioning (redundancy analysis) was applied to decompose salinity and contamination effects. The results indicated that cellular immune responses (total and differential haemocyte count, phagocytic activity and apoptosis) were mainly influenced by contaminants, whereas humoral factors (haemolytic activity) were mainly impacted by salinity. Hence, cellular immune functions may be suitable as biomarkers in monitoring programmes for the Baltic Sea and other geographic regions with salinity variances of the studied range.


Assessment of PRO label claims granted by the FDA as compared to the EMA (2006-2010).
The US Food and Drug Administration (FDA) provides formal guidance for the use of patient-reported outcomes (PROs) in support of labeling claims, whereas the European Medicines Agency (EMA) offers insight in a reflection paper relating to health-related quality of life in lieu of formal guidance. PRO label claims granted for new molecular entities and biologic license applications from 2006 through 2010 were reviewed to evaluate consistencies and discrepancies in PRO label claims granted by the FDA and the EMA and to highlight trends in the acceptance of PRO claims across agencies. Products approved by both the FDA and the EMA were identified. By using US Drug Approval Packages and European Public Assessment Reports packages, any PRO label claims made for the same product by the same company were compared. Both agencies approved a total of 75 products. Of these, 35 (47%) had at least one EMA-granted PRO label claim compared with 14 (19%) by the FDA. Most FDA-grated claims focused on symptoms; however, EMA-granted claims were more likely to include higher order concepts. Few (~12%) were granted the same label claims. Despite this discordance between the two agencies, where PRO label claims were granted by both the FDA and the EMA, there was similarity in the type of label claim. The EMA is more likely than the FDA to grant PRO claims and for higher order constructs. On a macro level, there appears to be poor concordance between claims granted by both agencies. On close examination, however, there appears to be greater concordance than previously recognized, which may be instructive in formulating future PRO strategies. Further research to create strategic alignment across agencies may be beneficial.


A New Standard for Crustacean Genomes: The Highly Contiguous, Annotated Genome Assembly of the Clam Shrimp Eulimnadia texana Reveals HOX Gene Order and Identifies the Sex Chromosome.
Vernal pool clam shrimp (Eulimnadia texana) are a promising model system due to their ease of lab culture, short generation time, modest sized genome, a somewhat rare stable androdioecious sex determination system, and a requirement to reproduce via desiccated diapaused eggs. We generated a highly contiguous genome assembly using 46× of PacBio long read data and 216× of Illumina short reads, and annotated using Illumina RNAseq obtained from adult males or hermaphrodites. Of the 120 Mb genome 85% is contained in the largest eight contigs, the smallest of which is 4.6 Mb. The assembly contains 98% of transcripts predicted via RNAseq. This assembly is qualitatively different from scaffolded Illumina assemblies: It is produced from long reads that contain sequence data along their entire length, and is thus gap free. The contiguity of the assembly allows us to order the HOX genes within the genome, identifying two loci that contain HOX gene orthologs, and which approximately maintain the order observed in other arthropods. We identified a partial duplication of the Antennapedia complex adjacent to the few genes homologous to the Bithorax locus. Because the sex chromosome of an androdioecious species is of special interest, we used existing allozyme and microsatellite markers to identify the E. texana sex chromosome, and find that it comprises nearly half of the genome of this species. Linkage patterns indicate that recombination is extremely rare and perhaps absent in hermaphrodites, and as a result the location of the sex determining locus will be difficult to refine using recombination mapping.


2,4 DNP improves motor function, preserves medium spiny neuronal identity, and reduces oxidative stress in a mouse model of Huntington's disease.
Huntington's disease (HD) is a neurodegenerative disorder caused by a CAG repeat expansion in the first exon of the gene huntingtin. There is no treatment to prevent or delay the disease course of HD currently. Oxidative stress and mitochondrial dysfunction have emerged as key determinants of the disease progression in HD. Therefore, counteracting mutant huntingtin (mHtt)-induced oxidative stress and mitochondrial dysfunction appears as a new approach to treat this devastating disease. Interestingly, mild mitochondrial uncoupling improves neuronal resistance to stress and facilitates neuronal survival. Mild mitochondrial uncoupling can be induced by the proper dose of 2,4-dinitrophenol (DNP), a proton ionophore that was previously used for weight loss. In this study, we evaluated the effects of chronic administration of DNP at three doses (0.5, 1, 5mg/kg/day) on mHtt-induced behavioral deficits and cellular abnormalities in the N171-82Q HD mouse model. DNP at a low dose (1mg/kg/day) significantly improved motor function and preserved medium spiny neuronal marker DARPP32 and postsynaptic protein PSD95 in the striatum of HD mice. Further mechanistic study suggests that DNP at this dose reduced oxidative stress in HD mice, which was indicated by reduced levels of F2-isoprostanes in the brain of HD mice treated with DNP. Our data indicated that DNP provided behavioral benefit and neuroprotective effect at a weight neutral dose in HD mice, suggesting that the potential value of repositioning DNP to HD treatment is warranted in well-controlled clinical trials in HD.


Alpha beta TCRs differ in the degree of their specificity for the positively selecting MHC/peptide ligand.
We have tested the peptide specificity of positive selection using three transgenic alphabetaTCRs, originally selected on class II MHC (A(b)) covalently bound with one peptide Ealpha (52-68) (Ep). The transgenic TCR specific for the cytochrome c-derived (43-58) peptide was selected on A(b) bound with different arrays of endogenous peptides or the analogue of Ep covalently bound to A(b), but not on the original A(b)Ep complex. In contrast, transgenic TCRs specific for two different analogues of the Ep peptide and A(b) did not mature as CD4(+) T cells in various thymic environments, including the A(b)EpIi(-) mice. These results show that TCRs can be promiscuous or specific for the selecting MHC/peptide complex, and suggest that in mice described in this study transgenic expression of the TCR changes the original requirements for the positively selecting MHC/peptide complex. Future studies will determine whether the latter phenomenon is general or specific for this system.


Matrix metalloproteinase-1 and tissue inhibitors do not predict incident coronary artery disease in the atherosclerosis risk in communities (ARIC) study.
Matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) are thought to be associated with coronary artery disease. The association of these markers with incident coronary artery disease has not been well described. Using a case-cohort design, we selected 216 individuals who had incident coronary artery disease (case group) and 225 individuals from a cohort random sample (comparison group) from participants enrolled in the Atherosclerosis Risk in Communities study. We measured plasma levels of MMP-1 and TIMP-1, traditional risk factors, and other markers of inflammation. We found no significant difference in TIMP-1 levels between the case group (827.8 +/- 23.8 ng/mL) and the comparison group (819.31 +/- 16.1 ng/mL) (P=0.77), and no significant difference in the frequency of MMP-1 levels that were dichotomized at the minimum detectable value of 1.7 ng/mL (P=0.49). In models adjusted for age, sex, race, body mass index, hypertension, diabetes, total cholesterol, high-density lipoprotein cholesterol, triglycerides, fibrinogen, von Willebrand factor, and white blood cell count, the hazard-rate ratio for incident coronary artery disease was 1.14 (95% confidence interval, 0.63-2.04; P=0.67) for individuals whose TIMP-1 levels were above, versus at or below the mean, and 1.17 (95% confidence interval, 0.63-2.19; P=0.62) for individuals whose MMP-1 levels were above 1.7 ng/mL. We conclude that TIMP-1 and MMP-1 levels in plasma were not predictive of incident coronary artery disease in a case-cohort random sample of the Atherosclerosis Risk in Communities study, a population study of asymptomatic middle-aged adults who had no prevalent atherosclerosis upon enrollment.


Brucella melitensis cell envelope protein and lipopolysaccharide epitopes involved in humoral immune responses of naturally and experimentally infected sheep.
Cell envelope fraction (CEF) of Brucella melitensis B115 was used to investigate antibody responses of B. melitensis naturally and strain H38 experimentally infected sheep by immunoblotting, indirect enzyme-linked immunosorbent assay (ELISA) (I-ELISA), and competitive ELISA (C-ELISA) with monoclonal antibodies (MAbs). MAbs used were directed to outer membrane proteins with molecular masses of 10, 16.5, 19, 25 to 27, 31 to 34, 36 to 38, and 89 kDa; to the heat shock protein DnaK, to O-polysaccharide (O-PS) common (C) and M epitopes; and to rough lipopolysaccharide (R-LPS) epitopes. In immunoblotting, all infected sheep sera tested recognized a large number of protein bands, including the above-cited proteins and other proteins for which MAbs have not been defined. The antibody response pattern was different from one animal to another, even within the experimentally infected sheep which were infected under the same experimental conditions. A number of protein bands were recognized by the sheep sera prior to experimental infection and by other uninfected sheep sera. The antibody reactivity to these antigens and others might explain the nonspecific antibody reactivity of sera in I-ELISA with CEF. C-ELISA confirmed also the individual variability of the antibody responses of infected sheep to protein antigens. Antibody responses to O-PS C and M epitopes were detected in all experimentally infected sheep and in half of the naturally infected sheep, but these responses were also heterogeneous in relation to their intensities. Antibody responses to R-LPS epitopes detected by use of C-ELISA with the anti-R-LPS MAbs were low or negative in most of the infected animals. Despite antibody response heterogeneity for CEF antigens, immunoblot and C-ELISA results indicated that, among the CEF antigens, the O-PS epitopes (C and M epitopes) and epitopes of the major 25- to 27- and 31- to 34-kDa outer membrane proteins seem to be the most promising for detecting B. melitensis infection in sheep.


Global trade in exotic pets 2006-2012.
International trade in exotic pets is an important and increasing driver of biodiversity loss and often compromises the standards required for good animal welfare. We systematically reviewed the scientific and gray literature and used the United Nations Environment Programme - World Conservation Monitoring Centre (UNEP-WCMC) Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) trade database to establish temporal and geographical trade patterns of live exotic birds, mammals, and reptiles and to describe trends in research, taxonomic representation, and level of threat and legal protection of species traded. Birds were the most species-rich and abundant class reported in trade; reptiles were second most abundant but unusually the most studied in this context; and mammals were least abundant in trade. Mammalian and reptilian species traded as pets were more likely to be threatened than expected by random. There have been a substantial number of Appendix I listed captive-bred mammals and birds and wild-caught birds and reptiles reported in trade to CITES. We identified the Middle East's emerging role as a driver of demand for exotic pets of all taxa alongside the well-established and increasing role of South America and Southeast Asia in the market. Europe, North America, and the Middle East featured most heavily in trade reports to CITES, whereas trade involving South America and Southeast Asia were given most emphasis in the literature. For effective monitoring of and appropriate response to the international exotic pet trade, it is imperative that the reliability and detail of CITES trade reports improve and that scientific research be directed toward those taxa and locations that are most vulnerable.


Associations of Thigh and Abdominal Adipose Tissue Radiodensity with Glucose and Insulin in Nondiabetic African-Ancestry Men.
Decreased radiodensity of adipose tissue (AT) located in the visceral AT (VAT), subcutaneous AT (SAT), and intermuscular AT (IMAT) abdominal depots is associated with hyperglycemia, hyperinsulinemia, and insulin resistance independent of AT volumes. These associations were sought in African-ancestry men, who have higher risk for type 2 diabetes and have been underrepresented in previous studies. This cross-sectional analysis included 505 nondiabetic men of African-Caribbean ancestry (median age: 61 years; median BMI: 26.8 kg/m2 ) from the Tobago Health Study. AT volumes and radiodensities were assessed using computed tomography, including abdominal (VAT and SAT) and thigh (IMAT) depots. Associations between AT radiodensities were assessed with fasting serum glucose and insulin and with insulin resistance (updated homeostatic model assessment of insulin resistance, HOMA2-IR). Higher radiodensity in any AT depot was associated with lower log-insulin and log-HOMA2-IR (β range: -0.16 to -0.18 for each; all P < 0.0001). No AT radiodensity was associated with glucose. Thigh IMAT radiodensity associations were independent of, and similar in magnitude to, VAT radiodensities. Model fit statistics suggested that AT radiodensities were a better predictor for insulin and insulin resistance compared with AT volumes in individuals with overweight and obesity. AT radiodensities at multiple depots are significantly associated with insulin and insulin resistance in African-ancestry men.


Development and validation of a modified quick SOFA scale for risk assessment in sepsis syndrome.
Sepsis is a severe clinical syndrome owing to its high mortality. Quick Sequential Organ Failure Assessment (qSOFA) score has been proposed for the prediction of fatal outcomes in sepsis syndrome in emergency departments. Due to the low predictive performance of the qSOFA score, we propose a modification to the score by adding age. We conducted a multicenter, retrospective cohort study among regional referral centers from various regions of the country. Participants recruited data of patients admitted to emergency departments and obtained a diagnosis of sepsis syndrome. Crude in-hospital mortality was the primary endpoint. A generalized mixed-effects model with random intercepts produced estimates for adverse outcomes. Model-based recursive partitioning demonstrated the effects and thresholds of significant covariates. Scores were internally validated. The H measure compared performances of scores. A total of 580 patients from 22 centers were included for further analysis. Stages of sepsis, age, time to antibiotics, and administration of carbapenem for empirical treatment were entered the final model. Among these, severe sepsis (OR, 4.40; CIs, 2.35-8.21), septic shock (OR, 8.78; CIs, 4.37-17.66), age (OR, 1.03; CIs, 1.02-1.05) and time to antibiotics (OR, 1.05; CIs, 1.01-1.10) were significantly associated with fatal outcomes. A decision tree demonstrated the thresholds for age. We modified the quick Sequential Organ Failure Assessment (mod-qSOFA) score by adding age (> 50 years old = one point) and compared this to the conventional score. H-measures for qSOFA and mod-qSOFA were found to be 0.11 and 0.14, respectively, whereas AUCs of both scores were 0.64. We propose the use of the modified qSOFA score for early risk assessment among sepsis patients for improved triage and management of this fatal syndrome.


Electrogram polarity and cavotricuspid isthmus block during ablation of typical atrial flutter.
The atrial activation sequence around the tricuspid annulus has been used to assess whether complete block has been achieved across the cavotricuspid isthmus during radiofrequency ablation of typical atrial flutter. However, sometimes the atrial activation sequence does not clearly establish the presence or absence of complete block. The purpose of this study was to determine whether a change in the polarity of atrial electrograms recorded near the ablation line is an accurate indicator of complete isthmus block. Radiofrequency ablation was performed in 34 men and 10 women (age 60 +/- 13 years [mean +/- SD]) with isthmus-dependent, counterclockwise atrial flutter. Electrograms were recorded around the tricuspid annulus using a duodecapolar halo catheter. Electrograms recorded from two distal electrode pairs (E1 and E2) positioned just anterior to the ablation line were analyzed during atrial flutter and during coronary sinus pacing, before and after ablation. Complete isthmus block was verified by the presence of widely split double electrograms along the entire ablation line. Complete bidirectional isthmus block was achieved in 39 (89%) of 44 patients. Before ablation, the initial polarity of E1 and E2 was predominantly negative during atrial flutter and predominantly positive during coronary sinus pacing. During incomplete isthmus block, the electrogram polarity became reversed either only at E2, or at neither E1 nor E2. In every patient, the polarity of E1 and E2 became negative during coronary sinus pacing only after complete isthmus block was achieved. In 4 patients (10%), the atrial activation sequence recorded with the halo catheter was consistent with complete isthmus block, but the presence of incomplete block was accurately detected by inspection of the polarity of E1 and E2. Reversal of polarity in bipolar electrograms recorded just anterior to the line of isthmus block during coronary sinus pacing after ablation of atrial flutter is a simple, quick, and accurate indicator of complete isthmus block.


Ethical aspects in medical publishing in Belgium.
Medical publishing has become a pivotal aspect of all stages of career development in medicine in Belgium. Publication pressure is therefore present from the stage of the medical student to the level of the head of an academic medical service. This pressure represents a real danger to commit scientific misconduct, ranging from minor missteps to severe fraud. Recent surveys have shown that scientific misconduct and publication fraud represent an important problem in medical publishing, more than in other scientific fields, also in Belgium. Several factors may explain the increased risk of scientific misconduct in medical publishing. Recent national and international initiatives have emerged in order to better understand the problem of scientific misconduct and to optimize ethics in medical publishing. They aim to increase integrity and transparency at all levels of medical research and publishing, starting at the beginning of a career in medical school. The current review describes the ethical difficulties in medical publishing in Belgium, and offers possible solutions.


A comparison of MRI criteria for diagnosing pediatric ADEM and MS.
Brain MRI is a useful tool for diagnosing inflammatory demyelinating disorders in children. However, it remains unclear which are the most reliable criteria for distinguishing multiple sclerosis (MS) from monophasic disorders such as acute disseminated encephalomyelitis (ADEM). We therefore compared the 4 current sets of MRI criteria in our Dutch pediatric cohort and determined which are the most useful in clinical practice for distinguishing ADEM from MS. We included 49 children who had had a demyelinating event and an MRI scan within 2 months of their first clinical attack. Twenty-one patients had ADEM and remained relapse-free after at least 2 years of follow-up. Twenty-eight patients had a definitive diagnosis of MS. We assessed the sensitivity and specificity of the following MRI criteria: Barkhof criteria, KIDMUS criteria, Callen MS-ADEM criteria, and Callen diagnostic MS criteria. The Callen MS-ADEM criteria had the best combination of sensitivity (75%) and specificity (95%). The KIDMUS criteria had higher specificity (100%), but much lower sensitivity (11%). The Barkhof criteria had a sensitivity of 61% and a specificity of 91%. The Callen diagnostic MS criteria were the most sensitive (82%), but were only 52% specific for distinguishing a first attack of MS from ADEM. The results in our cohort demonstrate that the new Callen criteria for multiple sclerosis-acute disseminated encephalomyelitis (MS-ADEM) are the most useful for differentiating a first attack of MS from monophasic ADEM. Although the Callen diagnostic MS criteria are more sensitive, they lack the specificity necessary to differentiate MS from ADEM.


Liver fibrosis after antiretroviral therapy in a longitudinal cohort of sexually infected HIV patients in eastern China.
We assessed the factors that influenced improvement or progression in human immunodeficiency virus (HIV)-infected patients who were receiving combination antiretroviral therapy (cART). This was a retrospective cohort study of HIV-infected patients receiving cART in Taizhou, Zhejiang, China, 2009-2015. Liver fibrosis was assessed by Fibrosis-4 (FIB-4) score. Improvement of liver fibrosis was defined as having > 30% decrease in FIB-4 from baseline, whereas progression of liver fibrosis was defined as having > 30% increase in FIB-4 score from baseline. A total of 955 HIV-infected patients were included. Of these, 808 (84.6%) were HIV-monoinfection, 125 (13.1%) were HIV/hepatitis B virus (HBV) coinfection and 29 (3.0%) were HIV/hepatitis C virus (HCV) coinfection. The median duration of treatment was 15 months. After treatment, 37.1% participants had > 30% decreases in FIB-4 index, 14.8% had > 30% increases in FIB-4 index, while the remaining 48.2% had stabilized FIB-4 index. In multivariate analysis, improvement of liver fibrosis was negatively associated with an older age, but was positively associated with baseline FIB-4 index and > 30% increases in CD4 cell count after ART. Progression of liver fibrosis was positively associated with an older age, but was negatively associated with gender and HIV transmission mode (male homosexual vs. male heterosexual, female heterosexual vs. male heterosexual), and baseline FIB-4 index. Our findings indicate that improvement of liver fibrosis could be achieved by early initiation of ART through better CD4 cell recovery. Liver fibrosis and hepatotoxicity associated with ART should be monitored as early as possible and throughout till the end of treatment, with special attention to the elderly and heterosexual men.


Phosphorylation and inactivation of glycogen synthase kinase 3 by protein kinase A.
Glycogen synthase kinase 3 (GSK-3) is implicated in multiple biological processes including metabolism, gene expression, cell fate determination, proliferation, and survival. GSK-3 activity is inhibited through phosphorylation of serine 21 in GSK-3 alpha and serine 9 in GSK-3 beta. These serine residues of GSK-3 have been previously identified as targets of protein kinase B (PKB/Akt), a serine/threonine kinase located downstream of phosphatidylinositol 3-kinase. Here, we show that serine 21 in GSK-3 alpha and serine 9 in GSK-3 beta are also physiological substrates of cAMP-dependent protein kinase A. Protein kinase A physically associates with, phosphorylates, and inactivates both isoforms of GSK-3. The results indicate that depending on the stimulatory context, the activity of GSK-3 can be modulated either by growth factors that work through the phosphatidylinositol 3-kinase-protein kinase B cascade or by hormonal stimulation of G protein-coupled receptors that link to changes in intracellular cAMP levels.


Perturbations in RhoA signalling cause altered migration and impaired neuritogenesis in human iPSC-derived neural cells with PARK2 mutation.
Mutations in parkin, encoded by the PARK2 gene, causes early-onset familial Parkinson's disease (PD), but dysfunctional parkin has also been implicated in sporadic PD. By combining human isogenic induced pluripotent stem cells (iPSCs) with and without PARK2 knockout (KO) and a novel large-scale mass spectrometry based proteomics and post-translational modification (PTM)-omics approach, we have mapped changes in protein profiles and PTMs caused by parkin deficiency in neurons. Our study identifies changes to several proteins previously shown to be dysregulated in brains of sporadic PD patients. Pathway analysis and subsequent in vitro assays reveal perturbations in migration and neurite outgrowth in the PARK2 KO neurons. We confirm the neurite defects using long-term engraftment of neurons in the striatum of immunosuppressed hemiparkinsonian adult rats. The GTP-binding protein RhoA was identified as a key upstream regulator, and RhoA activity was significantly increased in PARK2 KO neurons. By inhibiting RhoA signalling the migration and neurite outgrowth phenotypes could be rescued. Our study provides new insight into the pathogenesis of PD and demonstrates the broadly applicable potential of proteomics and PTMomics for elucidating the role of disease-causing mutations.


Real-time measurements of protein affinities on membrane surfaces by fluorescence spectroscopy.
Signal transduction in cells involves transitory interactions between proteins and membranes and between different proteins of the interacting species. These associations depend on the strength of the interactions and on the local concentration. Because the energy and intensity of the fluorescence of many probes are very sensitive to the local environment, fluorescence measurements can report on events, such as membrane binding and protein association, in real time. We describe methods to monitor associations both in vitro and in vivo by fluorescence.


Immunization alters body odor.
Infections have been shown to alter body odor. Because immune activation accompanies both infection and immunization, we tested the hypothesis that classical immunization might similarly result in the alteration of body odors detectable by trained biosensor mice. Using a Y-maze, we trained biosensor mice to distinguish between urine odors from rabies-vaccinated (RV) and unvaccinated control mice. RV-trained mice generalized this training to mice immunized with the equine West Nile virus (WNV) vaccine compared with urine of corresponding controls. These results suggest that there are similarities between body odors of mice immunized with these two vaccines. This conclusion was reinforced when mice could not be trained to directly discriminate between urine odors of RV- versus WNV-treated mice. Next, we trained biosensor mice to discriminate the urine odors of mice treated with lipopolysaccharide (LPS; a general elicitor of innate immunological responses) from the urine of control mice. These LPS-trained biosensors could distinguish between the odors of LPS-treated mouse urine and RV-treated mouse urine. Finally, biosensor mice trained to distinguish between the odors of RV-treated mouse urine and control mouse urine did not generalize this training to discriminate between the odors of LPS-treated mouse urine and control mouse urine. From these experiments, we conclude that: (1) immunization alters urine odor in similar ways for RV and WNV immunizations; and (2) immune activation with LPS also alters urine odor but in ways different from those of RV and WNV.


Stimulated Raman Scattering: From Bulk to Nano.
Stimulated Raman scattering (SRS) describes a family of techniques first discovered and developed in the 1960s. Whereas the nascent history of the technique is parallel to that of laser light sources, recent advances have spurred a resurgence in its use and development that has spanned across scientific fields and spatial scales. SRS is a nonlinear technique that probes the same vibrational modes of molecules that are seen in spontaneous Raman scattering. While spontaneous Raman scattering is an incoherent technique, SRS is a coherent process, and this fact provides several advantages over conventional Raman techniques, among which are much stronger signals and the ability to time-resolve the vibrational motions. Technological improvements in pulse generation and detection strategies have allowed SRS to probe increasingly smaller volumes and shorter time scales. This has enabled SRS research to move from its original domain, of probing bulk media, to imaging biological tissues and single cells at the micro scale, and, ultimately, to characterizing samples with subdiffraction resolution at the nanoscale. In this Review, we give an overview of the history of the technique, outline its basic properties, and present historical and current uses at multiple length scales to underline the utility of SRS to the molecular sciences.


Quantification of pleural effusion from single area measurements on CT.
The objective of this study was to determine if area measurements of pleural fluid on computed tomography (CT) reflect the actual pleural fluid volume (PEvol) as measured at autopsy, to establish a formula to estimate the volume of pleural effusion (PEest), and to test the accuracy and observer reliability of PEest.132 human cadavers, with pleural effusion were divided into phase 1 (n = 32) and phase 2 (n = 100). In phase 1, PEvol was compared to area measurements on axial (axA), sagittal (sagA), and coronal (corA) CT images. Linear regression analysis was used to create a formula to calculate PEest. In phase 2, intra-class correlation (ICC) was used to assess inter-reader reliability and determine the agreement between PEest and PEvol. PEvol correlated to a higher degree to axA (r s mean = 0.738; p < 0.001) than to sagA (r s mean = 0.679, p < 0.001) and corA (r s mean = 0.709; p < 0.001). PEest can be established with the following formula: axA × 0.1 = PEest. Mean difference between PEest and PEvol was less than 40 mL (ICC = 0.837-0.874; p < 0.001). Inter-reader reliability was higher between two experienced readers (ICC = 0.984-0.987; p < 0.001) than between an inexperienced reader and both experienced readers (ICC = 0.660-0.698; p < 0.001). Pleural effusions may be quantified in a rapid, reliable, and reasonably accurate fashion using single area measurements on CT.


Stem Cell Therapy as an Emerging Paradigm for Stroke (STEPS) II.
Cell-based therapies represent a new therapeutic approach for stroke. In 2007, investigators from academia, industry leaders, and members of the National Institutes of Health crafted recommendations to facilitate the translational development of cellular therapies as a novel, emerging modality for stroke from animal studies to clinical trials. This meeting was called Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS) and was modeled on the format of the Stroke Therapy Academic Industry Roundtable (STAIR) meetings. Since publication of the original STEPS guidelines, there has been an explosive growth in the number of cellular products and in the number of new laboratory discoveries that impact the safety and potential efficacy of cell therapies for stroke. Any successful development of a cell product will need to take into consideration several factors, including the preclinical safety and efficacy profile, cell characterization, delivery route, in vivo biodistribution, and mechanism of action. In 2010, a second meeting called STEPS 2 was held to bring together clinical and basic science researchers with industry, regulatory, and National Institutes of Health representatives. At this meeting, participants identified critical gaps in knowledge and research areas that require further studies, updated prior guidelines, and drafted new recommendations to create a framework to guide future investigations in cell-based therapies for stroke.


Review of the preclinical pharmacology and comparative efficacy of 5-hydroxytryptamine-3 receptor antagonists for chemotherapy-induced emesis.
This analysis was undertaken to review published reports of the comparative efficacy and safety of 5-hydroxytryptamine-3 (5-HT3) receptor antagonists in the prophylaxis of acute chemotherapy-induced emesis. Comparison data used are the preclinical pharmacology as well as the design and results of clinical trials. Seven comparative studies that used granisetron, ondansetron, or tropisetron in patients who received either moderately or highly emetogenic chemotherapy are reviewed. As the study designs, patient population, chemotherapy, antiemetic doses and schedule, and methods of assessment were slightly different, the results of each study are analyzed independently. Effectiveness is assessed by emetic episodes, nausea, and patient preference. The preclinical pharmacologic profile is different among the 5-HT3 antagonists in terms of potency, selectivity, dose response, and duration of action. The comparative clinical trials show that a single intravenous (i.v.) dose of granisetron 3 mg is as effective as multiple (8 mg x 3) or single (32 mg) i.v. doses of ondansetron for the prevention of acute nausea and emesis due to cisplatin. In the two moderately emetogenic clinical trials, granisetron 3 mg i.v. was at least as effective as ondansetron 8 mg i.v. +/- 24 mg orally and tropisetron 5 mg i.v. Patient preference was evaluated in three of the four crossover trials: granisetron was preferred in three of four, and no preference was reported in the fourth. The one trial to compare ondansetron 0.15 mg/kg x 3 versus granisetron 10 micrograms/kg x 1 or granisetron 40 micrograms/kg i.v. demonstrated equivalent control of nausea and vomiting in patients who received cisplatin-based chemotherapy. The 5-HT3 receptor antagonists compared are highly effective antiemetic agents that have now become the standard of care for preventing chemotherapy-induced emesis. Whether the described preclinical differences among these agents are also clinically significant remains to be seen. In the comparative trials analyzed, the 5-HT3 receptor antagonists demonstrated relatively equivalent clinical efficacy. Cost analysis may favor the use of one agent over another depending on the emetogenic challenge, dose of the 5-HT3 antagonists, and number of doses recommended. Patient preference may be an important factor to be considered in future antiemetic trials.


Racial and ethnic differences in ADHD and LD in young school-age children: parental reports in the National Health Interview Survey.
Racial and ethnic disparities have been documented for many physical health outcomes in children. Less is known, however, about disparities in behavioral and learning disorders in children. This study uses data from a national health survey to examine racial and ethnic differences in identified attention deficit hyperactivity disorder (ADHD) and learning disability (LD). The 1997-2001 National Health Interview Surveys obtained information from parents about the health and sociodemographic characteristics of children. Using these data, prevalence rates of identified ADHD and/or LD were estimated for Hispanic, African American, and white children 6-11 years of age. Racial and ethnic differences in health conditions, income, and insurance coverage were examined as possible explanations for disparities in parental reports of ADHD and LD, as well as the use of any prescription medication among children with ADHD. Hispanic and African American children, compared to white children, had parental reports of identified ADHD without LD less often, and adjustments for the confounding variables-birthweight, income, and insurance coverage-did not eliminate these differences. Hispanic and African American children, compared to white children, also had parental reports of ADHD with LD less often after adjustments for the effects of confounding variables. By contrast, after adjustments for confounding variables, Hispanic and African American children were as likely as white children to have LD without ADHD. Among children with ADHD, use of any prescription medication was reported less often for Hispanic and African American children than white children. These disparities in medication use persisted after adjustments for confounding variables. The prevalence of ADHD and the use of any prescription medication among children with ADHD differed among Hispanic, African American, and white children. These disparities could not be explained by racial and ethnic differences in other health conditions and sociodemographic variables.


Twenty-year trends in incidence and 1-year mortality in Swedish patients hospitalised with non-AMI chest pain. Data from 1987-2006 from the Swedish hospital and death registries.
To study trends for 20 years in incidence and 1-year mortality in hospitalised patients who received a diagnosis of either angina or unexplained chest pain (UCP) in Sweden. Register study of all patients aged 25-84 years identified from the Swedish National Hospital Discharge Register who were hospitalised with a first-time diagnosis of UCP or angina pectoris during 1987 to 2006. A total of 378 454 patients, 235 855 with UCP and 142 599 with angina. 1-Year mortality and standardised mortality ratios (SMRs). From the period 1987-1991 to 2002-2006, the observed 1-year mortality rate in men and women with UCP aged 25-74 years decreased from 2.19% to 1.45% and from 1.85% to 0.91%, respectively. SMRs decreased from 1.67 (95% CI 1.39 to 1.95) and 1.63 (1.27 to 2.00) to 1.09 (0.96 to 1.23) and 0.88 (0.75 to 1.00). Corresponding decreases in 1-year mortality for a discharge diagnosis of angina were from 6.50% to 2.49% in men and from 4.80% to 1.68% in women, with SMRs decreasing from 2.69 (2.33-3.05) and 2.59 (2.06-3.12) to 1.09 (0.93-1.25) and 1.05 (0.81-1.29), respectively. Similar changes occurred in patients aged 75-84 years. Only men with UCP aged 75-84 years still retained a slightly increased mortality (SMR 1.14 (1.01-1.28)). The prognosis of patients admitted with chest pain in which acute myocardial infarction has been ruled out has improved for the past 20 years, such that the 1-year mortality of these patients is now similar to that in the general population.


Initiation of warfarin therapy: comparison of physician dosing with computer-assisted dosing.
In a prospective, randomized study at two university hospitals, the authors examined how effectively housestaff physicians (n = 36) managed the initiation of warfarin therapy compared with a computer-assisted dosing regimen (n = 39) using the software program Warfcalc, which was managed by one of the authors. Target prothrombin time ratios were selected by the physicians. Study endpoints included: the time to reach a therapeutic prothrombin ratio, the time to reach a stable therapeutic dose, the number of patients transiently overanticoagulated, the number of bleeding complications, and the accuracy of the predicted maintenance dose, which was assessed at steady-state 10-14 days later. Computer-assisted dosing consistently out-performed the physicians: a stable therapeutic dose was achieved 3.7 days earlier (p = 0.002), fewer patients were overanticoagulated (10% versus 41%), and the predicted maintenance dose was in the therapeutic range in 85% of the computer-dosed patients versus 42% of the physician group (p less than 0.002). For physicians who did not routinely manage warfarin therapy, computer-assisted dosing improved the accuracy of dosing and shortened the time required to achieve a stable therapeutic dose.


Costs and outcome of assertive community treatment (ACT) in a rural area in Denmark: 4-year register-based follow-up.
Health economic evidence of assertive community treatment (ACT) in Denmark is limited. The aim of the study was to assess the costs and outcome of ACT among 174 patients with severe and persistent mental illness in a rural area of Denmark. The study was based on a quasi-experimental design with a control group from the neighbouring region. Costs and retention in mental health services were analysed by using register data 1 year before and 4 years after inclusion in the study. Data on the use of supportive housing were available for the year before baseline and the subsequent 2 years only. Seventy eight percent of the patients receiving ACT were in contact with psychiatric services at the 4-year follow-up, while 69% of the patients in the control group had contact with psychiatric services (P < 0.17). Days in supportive housing were lower for the ACT group before baseline and remained so (dropping to zero) for the subsequent 2 years. Over 4 years, the mean total costs per patient in the group receiving ACT were DDK 493,442 (SE = 34,292). Excluding costs of supportive housing, the mean total costs per patient of the control group were DDK 537,218 (SE = 59,371), P < 0.53. If these costs are included, however, the mean total costs for the ACT group are unchanged, whereas costs for the control group rise to DDK 671,500 (SE = 73,671), P < 0.03. While ACT appears to have resulted in a significant reduction in costs for psychiatric hospitalizations, baseline differences in use of supportive housing make the effects of ACT on overall costs more ambiguous. At worst, however, overall costs did not increase. Given the generally acknowledged clinical benefits of ACT over standard outpatient care, the results support further dissemination of ACT in Denmark.


Effects of nicotine on bone mass and strength in aged female rats.
This study investigated the effects of nicotine on bone mass and biomechanical properties in aged, estrogen-replete (sham-operated) and estrogen-deplete (ovariectomized) female rats. Eight month old, retired breeder, sham-operated and ovariectomized Sprague-Dawley rats were left untreated for 12 weeks to establish cancellous osteopenia in the ovariectomized group. The animals were then administered saline, low dose nicotine (6.0 mg/kg/day) or high dose nicotine (9.0 mg/kg/day) via osmotic minipumps for 12 weeks. Vertebrae and femora were collected at necropsy for determination of bone mass and strength. As expected, ovariectomy had a negative effect on most endpoints evaluated. Vertebral body bone mineral content (BMC) and density (BMD) and the structural (ultimate load and yield load) and material (ultimate stress, yield stress, and flexural modulus of elasticity) strength properties were lower in the OVX rats than in the sham-operated rats. Femoral diaphysis BMC, BMD, ultimate load, and flexural modulus were also lower in the OVX rats than in the sham-operated rats. The nicotine doses administered resulted in serum nicotine levels that averaged 1.5-4.5-fold greater than those observed in heavy smokers. Despite the high doses used, nicotine had no effect on vertebral BMC, BMD, or any of the structural and material strength properties in either the OVX or the Sham rats. In addition, nicotine had no effect on femoral diaphysis BMC, BMD, ultimate load, stiffness, ultimate stress, or flexural modulus. Femoral yield load and stress were lower in low dose nicotine-treated rats than in vehicle-treated rats. However, differences were not detected between the high dose nicotine- and vehicle-treated rats for either femoral yield load or stress. The results suggest that tobacco agents other than nicotine are responsible for the decreased bone density and increased fracture risk as observed in smokers.


Assessing nursing problems of importance for the development of nursing care in a radiation therapy department.
The challenges to nurses working in the rapidly changing milieu of radiotherapy in cancer care are great. New treatment regimens continually require reexamination of old assumptions about patients' needs and communication between colleagues. In order to continue the development of the nursing care in the radiation therapy department, the purpose of this study was to assess the nursing problems experienced in the care of the cancer patients and the nursing problems experienced in work with other professionals from the nurse's perspective. Eighteen of the 38 nurses in the radiation therapy department were invited to participate in the study. The mean age of the nurses was 39 years (range 25-57 years). The study was carried out by using a three-phase, structured, communication-process method according to the Delphi technique. The nurses reported "poor follow-up of patients who have completed radiation therapy," closely followed by "finding time to document the nursing care" and "finding time to treat the scheduled patients" as the most problematic areas of importance for the development of nursing care. The nursing problems considered to be the most difficult in the work with other professionals were "lack of communication," "lack of knowledge of our work and competence," and "lack of comprehension of each other's professions." With the present trend in the health care system toward smaller financial resources, it is necessary to find new ways of organizing nursing care in order to find time to take care of the patient's problems during and after the radiation therapy. This study constituted the base for organizing a nursing care clinic in the department with sufficient time to assess and monitor side effects, patient education, sexual counselling, and coping assistance.


The maize ID1 flowering time regulator is a zinc finger protein with novel DNA binding properties.
The INDETERMINATE protein, ID1, plays a key role in regulating the transition to flowering in maize. ID1 is the founding member of a plant-specific zinc finger protein family that is defined by a highly conserved amino sequence called the ID domain. The ID domain includes a cluster of three different types of zinc fingers separated from a fourth C2H2 finger by a long spacer; ID1 is distinct from other ID domain proteins by having a much longer spacer. In vitro DNA selection and amplification binding assays and DNA binding experiments showed that ID1 binds selectively to an 11 bp consensus motif via the ID domain. Unexpectedly, site-directed mutagenesis of the ID1 protein showed that zinc fingers located at each end of the ID domain are not required for binding to the consensus motif despite the fact that one of these zinc fingers is a canonical C2H2 DNA binding domain. In addition, an ID1 in vitro deletion mutant that lacks the extra spacer between zinc fingers binds the same 11 bp motif as normal ID1, suggesting that all ID domain-containing proteins recognize the same DNA target sequence. Our results demonstrate that maize ID1 and ID domain proteins have novel zinc finger configurations with unique DNA binding properties.


Osteotomy, arthrodesis, and arthroplasty for complex multiapical deformity of the leg.
Assessment of diaphyseal deformity in the tibia consists of delineating anatomic axes or cortical lines with resultant apices of deformity. Single-apex deformities have been well described both in terms of assessment and treatment, whereas double-level deformities with metaphyseal and/or juxtacortical involvement are less straightforward. Multiapical deformities of the lower extremity, though uncommon, are the next level of complexity and provide the surgeon with a difficult correctional mission. We report two cases of multiapical deformity of the tibia, both of which were secondary to a distant history of tibial fracture with resultant knee and ankle joint degeneration and deformity. Both cases had three levels of deformity that were addressed with tibial osteotomy, ankle fusion, and total knee replacement. Initial presentation, problem lists, surgical treatment, and subsequent results are reviewed. Treatment of a post-traumatic three-level deformity has never specifically been addressed in the literature, although the principles of treatment are the same as for less complex deformities. These two cases present a treatment approach for complex, multiapical deformity of the tibia. The same principles of deformity correction used to treat less complex deformities are applied to these patients with an overarching synthesis that takes all aspects of the three deformities into account. Although these cases are complex and difficult, good results in terms of deformity correction and pain relief can be obtained.


Determining sample size for the morphological assessment of sperm.
Morphologic assessment of spermatozoa is an integral component in the analysis of semen. Whether a technician rapidly screening semen quality at a commercial stud, a veterinarian performing breeding soundness examinations, a clinician at a reference andrology laboratory providing auditing or diagnostic services, or a researcher evaluating morphology as a part of a fertility study, it is important to make an informed decision regarding the number of spermatozoa to include in the morphology assessment. Application of basic statistical principles such as the nature of proportions, level of confidence in an observed value, and the interaction of sample size with precision, can and should be used in the decision process. This paper outlines in detail the application of these statistical principles in relation to the morphologic assessment of spermatozoa. Guidelines on how these principles can be utilized in practical situations are discussed. Additionally, methodologies for comparison of results within and between laboratories (an area easily prone to misinterpretation) are reviewed. It is hoped that through the use of these fundamental statistical principles, this paper will bring clarity and delineation to the science of quantifying the morphology of spermatozoa.


How People with Low Vision Achieve Magnification in Digital Reading.
Digital reading displays provide opportunities for enhancing accessibility of text for low vision. How are these displays used by people in their daily lives? Subjects responded to an online survey concerning their vision history, reading technology, display preferences, and reading habits. Here, we report on findings concerning acuity and magnification. The survey asked subjects to arrange a text passage for typical reading and to report viewing distance, screen dimensions, and the number of characters per line. Seventy-five adult subjects (most with early-onset low vision, few with central field loss) completed all survey questions relevant to the analysis of acuity and magnification. Mean acuity was .92 logMAR (range, 0.1 to 1.6), and mean age was 44.8 years (range, 18 to 71 years). Twelve normally sighted controls reported the same information while viewing the passage on cell phones, tablets, and computers. The controls had a mean viewing distance of 38.7 cm and a mean x-height of 1.38 mm. For all three types of devices, angular x-height was 0.21° (close to laboratory estimates of the critical print size for reading). Low vision subjects showed decreasing viewing distance and increasing print size with larger values of logMAR acuity. Most of the low vision subjects achieved their desirable magnification by a combination of reduced viewing distance and increased physical letter size. The majority (54 of 75) relied more on letter-size magnification. Relative to the controls, regression analysis revealed that a typical low vision subject with logMAR acuity of 1.0 reduced viewing distance by a factor of 2.8 and enlarged physical print size by a factor of 6. Our survey shows that people with a wide range of acuities are engaged in digital reading. Our subjects achieved desirable magnification primarily by enlarging physical character size and to a lesser extent by reducing viewing distance.