Mesh versus non-mesh for inguinal and femoral hernia repair.
This is an update of a Cochrane Review first published in 2001.Hernias are protrusions of all or part of an organ through the body wall that normally contains it. Groin hernias include inguinal (96%) and femoral (4%) hernias, and are often symptomatic with discomfort. They are extremely common, with an estimated lifetime risk in men of 27%. Occasionally they may present as emergencies with complications such as bowel incarceration, obstruction and strangulation. The definitive treatment of all hernias is surgical repair, inguinal hernia repair being one of the most common surgical procedures performed. Mesh (hernioplasty) and the traditional non-mesh repairs (herniorrhaphy) are commonly used, with an increasing preference towards mesh repairs in high-income countries. To evaluate the benefits and harms of different inguinal and femoral hernia repair techniques in adults, specifically comparing closure with mesh versus without mesh. Outcomes include hernia recurrence, complications (including neurovascular or visceral injury, haematoma, seroma, testicular injury, infection, postoperative pain), mortality, duration of operation, postoperative hospital stay and time to return to activities of daily living. We searched the following databases on 9 May 2018: Cochrane Colorectal Cancer Group Specialized Register, Cochrane Central Register of Controlled Trials (Issue 1), Ovid MEDLINE (from 1950), Ovid Embase (from 1974) and Web of Science (from 1900). Furthermore, we checked the WHO International Clinical Trials Registry Platform (ICTRP) and ClinicalTrials.gov for trials. We applied no language or publication restrictions. We also searched the reference lists of included trials and review articles. We included randomised controlled trials of mesh compared to non-mesh inguinal or femoral hernia repairs in adults over the age of 18 years. We used standard methodological procedures expected by Cochrane. Where available, we collected information on adverse effects. We presented dichotomous data as risk ratios, and where possible we calculated the number needed to treat for an additional beneficial outcome (NNTB). We presented continuous data as mean difference. Analysis of missing data was based on intention-to-treat principles, and we assessed heterogeneity using an evaluation of clinical and methodological diversity, Chi2 test and I2 statistic. We used GRADE to assess the quality of evidence for each outcome. We included 25 studies (6293 participants) in this review. All included studies specified inguinal hernias, and two studies reported that femoral hernias were included.Mesh repair probably reduces the risk of hernia recurrence compared to non-mesh repair (21 studies, 5575 participants; RR 0.46, 95% CI 0.26 to 0.80, I2 = 44%, moderate-quality evidence). In absolute numbers, one hernia recurrence was prevented for every 46 mesh repairs compared with non-mesh repairs. Twenty-four studies (6293 participants) assessed a wide range of complications with varying follow-up times. Neurovascular and visceral injuries were more common in non-mesh repair groups (RR 0.61, 95% CI 0.49 to 0.76, I2 = 0%, NNTB = 22, high-quality evidence). Wound infection was found slightly more commonly in the mesh group (20 studies, 4540 participants; RR 1.29, 95% CI 0.89 to 1.86, I2 = 0%, NNTB = 200, low-quality evidence). Mesh repair reduced the risk of haematoma compared to non-mesh repair (15 studies, 3773 participants; RR 0.88, 95% CI 0.68 to 1.13, I2 = 0%, NNTB = 143, low-quality evidence). Seromas probably occur more frequently with mesh repair than with non-mesh repair (14 studies, 2640 participants; RR 1.63, 95% CI 1.03 to 2.59, I2 = 0%, NNTB = 72, moderate-quality evidence), as does wound swelling (two studies, 388 participants; RR 4.56, 95% CI 1.02 to 20.48, I2 = 33%, NNTB = 72, moderate-quality evidence). The comparative effect on wound dehiscence is uncertain due to wide confidence intervals (two studies, 329 participants; RR 0.55, 95% CI 0.12 to 2.48, I2 = 37% NNTB = 77, low-quality evidence). Testicular complications showed nearly equivocal results; they probably occurred slightly more often in the mesh group however the confidence interval around the effect was wide (14 studies, 3741 participants; RR 1.06, 95% CI 0.63 to 1.76, I2 = 0%, NNTB = 2000, low-quality evidence). Mesh reduced the risk of postoperative urinary retention compared to non-mesh (eight studies, 1539 participants; RR 0.53, 95% CI 0.38 to 0.73, I2 = 56%, NNTB = 16, moderate-quality evidence).Postoperative and chronic pain could not be compared due to variations in measurement methods and follow-up time (low-quality evidence).No deaths occurred during the follow-up periods reported in the seven studies (2546 participants) reporting this outcome (high-quality evidence).The average operating time was longer for non-mesh repairs by a mean of 4 minutes 22 seconds, despite wide variation across the studies regarding size and direction of effect, thus this result is uncertain (20 studies, 4148 participants; 95% CI -6.85 to -1.60, I2= 97%, very low-quality evidence). Hospital stay may be shorter with mesh repair, by 0.6 days (12 studies, 2966 participants; 95% CI -0.86 to -0.34, I2 = 98%, low-quality evidence), and participants undergoing mesh repairs may return to normal activities of daily living a mean of 2.87 days sooner than those with non-mesh repair (10 studies, 3183 participants; 95% CI -4.42 to -1.32, I2 = 96%, low-quality evidence), although the results of both these outcomes are also limited by wide variation in the size and direction of effect across the studies. Mesh and non-mesh repairs are effective surgical approaches in treating hernias, each demonstrating benefits in different areas. Compared to non-mesh repairs, mesh repairs probably reduce the rate of hernia recurrence, and reduce visceral or neurovascular injuries, making mesh repair a common repair approach. Mesh repairs may result in a reduced length of hospital stay and time to return to activities of daily living, but these results are uncertain due to variation in the results of the studies. Non-mesh repair is less likely to cause seroma formation and has been favoured in low-income countries due to low cost and reduced availability of mesh materials. Risk of bias in the included studies was low to moderate and generally handled well by study authors, with attention to details of allocation, blinding, attrition and reporting.


Quantitative fluorescence loss in photobleaching for analysis of protein transport and aggregation.
Fluorescence loss in photobleaching (FLIP) is a widely used imaging technique, which provides information about protein dynamics in various cellular regions. In FLIP, a small cellular region is repeatedly illuminated by an intense laser pulse, while images are taken with reduced laser power with a time lag between the bleaches. Despite its popularity, tools are lacking for quantitative analysis of FLIP experiments. Typically, the user defines regions of interest (ROIs) for further analysis which is subjective and does not allow for comparing different cells and experimental settings. We present two complementary methods to detect and quantify protein transport and aggregation in living cells from FLIP image series. In the first approach, a stretched exponential (StrExp) function is fitted to fluorescence loss (FL) inside and outside the bleached region. We show by reaction-diffusion simulations, that the StrExp function can describe both, binding/barrier-limited and diffusion-limited FL kinetics. By pixel-wise regression of that function to FL kinetics of enhanced green fluorescent protein (eGFP), we determined in a user-unbiased manner from which cellular regions eGFP can be replenished in the bleached area. Spatial variation in the parameters calculated from the StrExp function allow for detecting diffusion barriers for eGFP in the nucleus and cytoplasm of living cells. Polyglutamine (polyQ) disease proteins like mutant huntingtin (mtHtt) can form large aggregates called inclusion bodies (IB's). The second method combines single particle tracking with multi-compartment modelling of FL kinetics in moving IB's to determine exchange rates of eGFP-tagged mtHtt protein (eGFP-mtHtt) between aggregates and the cytoplasm. This method is self-calibrating since it relates the FL inside and outside the bleached regions. It makes it therefore possible to compare release kinetics of eGFP-mtHtt between different cells and experiments. We present two complementary methods for quantitative analysis of FLIP experiments in living cells. They provide spatial maps of exchange dynamics and absolute binding parameters of fluorescent molecules to moving intracellular entities, respectively. Our methods should be of great value for quantitative studies of intracellular transport.


Instability of a plasmid-borne inverted repeat in Saccharomyces cerevisiae.
Inverted repeated DNA sequences are common in both prokaryotes and eukaryotes. We found that a plasmid-borne 94 base-pair inverted repeat (a perfect palindrome of 47 bp) containing a poly GT sequence is unstable in S. cerevisiae, with a minimal deletion frequency of about 10(-4)/mitotic division. Ten independent deletions had identical end points. Sequence analysis indicated that all deletions were the result of a DNA polymerase slippage event (or a recombination event) involving a 5-bp repeat (5' CGACG 3') that flanked the inverted repeat. The deletion rate and the types of deletions were unaffected by the rad52 mutation. Strains with the pms1 mutation had a 10-fold elevated frequency of instability of the inverted repeat. The types of sequence alterations observed in the pms1 background, however, were different than those seen in either the wild-type or rad52 genetic backgrounds.


Vedolizumab Induces Endoscopic and Histologic Remission in Patients With Crohn's Disease.
We evaluated the ability of vedolizumab to induce endoscopic and histologic remission in patients with Crohn's disease (CD). We performed a prospective study of 110 patients with active CD, based on CD activity index (CDAI) scores >220 and mucosal ulcerations, who received open-label vedolizumab (300 mg) infusions at weeks 0, 2, and 6, and every 8 weeks thereafter through week 52 at tertiary centers in Europe. Patients received an additional infusion at week 10 if their CDAI score had not decreased by 70 points. Patients underwent ileocolonoscopy with collection of biopsies at baseline and weeks 26 and 52; a local and central reader determined simple endoscopic index for CD (SES-CD) scores. Histologic features were assessed by a blinded pathologist at week 26. Serum concentrations of vedolizumab were measured at serial time points. The primary outcome was endoscopic and histologic remission in patients with active CD treated with vedolizumab for 52 weeks. At weeks 26 and 52, 36 patients (29%) and 34 patients (31%), respectively, were in corticosteroid-free clinical remission (CDAI score <150), respectively. Based on intent-to-treat analysis, endoscopic remission (SES-CD score <4) was achieved by 36 patients (33%) and 40 patients (36%) at weeks 26 and 52. Endoscopic responses (decrease in SES-CD score ≥50%) occurred in 44 patients (40%) at week 26 and 5 patients (45%) at week 52. Serum concentrations of vedolizumab were higher at weeks 2, 10, and 22 in patients with lower SES-CD scores. Histologic remission at week 26 was observed in 43 (64%) of 67 patients based on Geboes Score and 37 (66%) of 56 patients based on Robarts Histopathology Index scores in analyses of paired biopsies with inflammation at baseline. Serum concentrations of vedolizumab above 10 mg/L at week 22 were associated with endoscopic remission at week 26. In a prospective trial, we found that approximately one-third of patients with CD achieve endoscopic remission after 52 weeks of treatment with vedolizumab and two-thirds achieve histologic remission at week 26. Higher serum concentrations of vedolizumab were associated with better outcomes. EUDRACT no: 2014-005376-29.


SU-E-T-180: The Radiological Physics Center's Anthropomorphic Quality Assurance Phantom Program.
To describe the phantoms, program logistics and current results for the Radiological Physics Center's (RPC) anthropomorphic QA phantom program for credentialing institutions for participation in NCI-sponsored advanced technology clinical trials. The RPC has developed an extensive phantom credentialing program consisting of four different phantoms designs: H&N, pelvis, lung and spine. These QA phantoms are water-filled plastic shells with imageable targets, avoidance structures, and heterogeneities that contain TLD and radiochromic film dosimeters. Institutions wishing to be credentialed request a phantom and are prioritized for delivery. At the institution, the phantom is imaged, a treatment plan is developed, the phantom is positioned on the treatment couch and the treatment is delivered. The phantom is returned and the measured dose distributions are compared to the institution's electronically submitted treatment plan dosimetry data. The RPC currently has an inventory of 31 H&N, 10 pelvis, 9 lung, and 8 spine phantoms that are mailed to institutions nationally and internationally. In 2011, 444 of these phantoms were mailed out for credentialing. Once the phantom is sent, it takes the institution an average of 26 days to return it to the RPC. On average the dosimeters are analyzed within 17 days and the report is sent 21 days after receipt of the phantom data. In 2011 the percent of phantoms meeting the acceptance criteria increased by 12, 13 and 6 percentage points for the H&N, spine and lung phantoms, respectively. It fell by 5 percentage points for the pelvis phantom. The RPC's QA phantom program has been an effective and responsive QA tool for assessing the use of advanced technologies in NCI sponsored clinical trials. The RPC has been efficient in its mailing of phantoms, and analyzing and reporting results. Work supported by PHS grant CA10953 and CA081647 (NCI, DHHS).


Diet composition is associated with endogenous formation of N-nitroso compounds in obese men.
Endogenous formation of carcinogenic N-nitroso compounds (NOC) occurs in the human gut. Red meat is considered the most important dietary component linked to NOC formation, although nitrate and vitamin C (VitC) also contribute. We previously showed that high-protein weight-loss diets increased fecal NOC and this was enhanced by simultaneous carbohydrate restriction. Although previous studies have focused on the effect of either 1 or 2 dietary components on endogenous NOC formation, no study to date has investigated the combined contribution of various dietary components. The current study therefore assessed the joint impact of several known dietary contributors to the endogenous formation of NOC in obese men. It also aimed to identify further novel contributors and investigate their role in explaining shifts in endogenous formation of NOC. Three dietary trials were conducted in obese men consuming body weight maintenance or weight-loss diets, with NOC measured in fecal samples. Consumption of meat-based weight-loss diets increased (P < 0.001) fecal NOC. Red meat intake was positively correlated with the fecal log NOC concentration (r = 0.60; P < 0.001). Dietary carbohydrate and sugar were negatively correlated with the fecal log NOC concentration (r = -0.66 for both; P < 0.001). Multiple regression analysis identified several dietary components that drive endogenous NOC formation, namely, red meat, nitrate, VitC, total energy, and nonstarch polysaccharides. We present a regression model that predicts endogenous NOC formation in obese men based on their dietary intakes. This model could improve the estimation of endogenous NOC formation, currently used in epidemiological studies into diet and cancer.


The Cumulative Prevalence of Termination of Parental Rights for U.S. Children, 2000-2016.
Recent research has used synthetic cohort life tables to show that having a Child Protective Services investigation, experiencing confirmed maltreatment, and being placed in foster care are more common for American children than would be expected based on daily or annual rates for these events. In this article, we extend this literature by using synthetic cohort life tables and data from the Adoption and Foster Care Analysis and Reporting System to generate the first cumulative prevalence estimates of termination of parental rights. The results provide support for four conclusions. First, according to the 2016 estimate, 1 in 100 U.S. children will experience the termination of parental rights by age 18. Second, the risk of experiencing this event is highest in the first few years of life. Third, risks are highest for Native American and African American children. Nearly 3.0% of Native American children and around 1.5% of African American children will ever experience this event. Finally, there is dramatic variation across states in the risk of experiencing this event and in racial/ethnic inequality in this risk. Taken together, these findings suggest that parental rights termination, which involves the permanent loss of access to children for parents, is far more common than often thought.


Content encapsulation in consciousness is likely to be incomplete.
Although the passive frame theory (PST) seems to be very plausible, the encapsulation of content generation deserves further elaboration. Many concepts and data considering the capacity theory, perception, and higher cognitive processes seem to contradict this principle. In general, PST can be suitable in many cases, but it needs to be completed by other mechanisms.


Skull fracture and the diagnosis of abuse.
Eighty nine children under 2 years of age with skull fracture were studied retrospectively--29 children with definite non-accidental injury serially recorded by the Departments of Paediatrics and Forensic Medicine, and 60 children consecutively admitted to hospital with skull fractures after accidents. There were 20 deaths including 19 among abused children. Multiple injuries and an inadequate history assisted in diagnosing abuse. Fracture characteristics found considerably more often in abused children were: multiple or complex configuration; depressed, wide, and growing fracture; involvement of more than a single cranial bone; non-parietal fracture; and associated intracranial injury including subdural haematoma. No fractures measuring more than 5.0 mm on presentation were found after accidents, but 6 of these 'growing fractures' were found in abused children. Accidents usually resulted in single, narrow, linear fractures most commonly of the parietal, with no associated intracranial injury. The results suggest that in skull fracture in young children where a minor fall is alleged, it is possible to recognise abuse by consideration of the fracture alone.


Long-range connectomics.
Decoding neural algorithms is one of the major goals of neuroscience. It is generally accepted that brain computations rely on the orchestration of neural activity at local scales, as well as across the brain through long-range connections. Understanding the relationship between brain activity and connectivity is therefore a prerequisite to cracking the neural code. In the past few decades, tremendous technological advances have been achieved in connectivity measurement techniques. We now possess a battery of tools to measure brain activity and connections at all available scales. A great source of excitement are the new in vivo tools that allow us to measure structural and functional connections noninvasively. Here, we discuss how these new technologies may contribute to deciphering the neural code.


Optofluidic SERS chip with plasmonic nanoprobes self-aligned along microfluidic channels.
This work reports an optofluidic SERS chip with plasmonic nanoprobes self-aligned along microfluidic channels. Plasmonic nanoprobes with rich electromagnetic hot spots are selectively patterned along PDMS microfluidic channels by using a Scotch tape removal and oxygen plasma treatment, which also provide the permanent bonding between PDMS and a glass substrate. A silver film with an initial thickness of 30 nm after oxygen plasma treatment creates nanotips and nanodots with a maximum SERS performance, which were successfully implanted with microfluidic concentration gradient generators. The novel device enables the label-free and solution-phase SERS detection of small molecules with low Raman activity such as dopamine at micromolar level in flow. This optofluidic SERS chip can be readily expanded for microfluidic networks with diverse functions for advanced optical biochemical assays.


Systemic onset juvenile idiopathic arthritis: a single center experience.
Sağ E, Uzunoğlu B, Bal F, Sönmez HE, Demir S, Bilginer Y, Özen S. Systemic onset juvenile idiopathic arthritis: a single center experience. Turk J Pediatr 2019; 61: 852-858. Systemic juvenile idiopathic arthritis (sJIA) presents with prolonged fever and systemic features such as arthritis, rash, lymphadenopathy, hepatosplenomegaly and serositis. In this study, we aimed to evaluate the clinical and laboratory findings, and outcomes of sJIA patients from a tertiary rheumatology center. Between 2010-2017, patients who had been diagnosed with sJIA, participated in the study. The demographics, clinical and laboratory features, and outcomes, were evaluated retrospectively. Seventy-five sJIA (%56 male) patients were enrolled. The mean age at diagnosis was 6,45±4,80 years. At the time of diagnosis, the most common findings were fever (%100) followed by arthritis (78,7%), and rash (66,2%). Twenty-four percent of the patients present with macrophage activation syndrome (MAS) at the time of diagnosis. Totally, 36% of the patients had at least one MAS attack during the course of the disease. 46% of the patients had polyphasic course while 54% had one attack (26% monophasic, 28% persistant). All of the patients were treated with non-steroid anti-inflammatory drugs (NSAID) and/or corticosteroids at the beginning of the disease. Twenty percent of the patients reached remission with corticosteroid or disease-modifying anti-rheumatic drugs (DMARDs) however the rest of the patients needed at least one biologic agent. Anakinra was the most common first-line biologic treatment choice (n=45). Fourteen (18,7%) of the patients had polyarticular joint involvement during the disease course, and 5 of them achieved remission with tocilizumab. Systemic JIA is an important disease with high risk of morbidity and mortality. As our center is one of the most important tertiary referral rheumatology centers in the country, we had a high MAS incidence. Eighty percent of the patients achieved remission with a biological agent. Anti-IL1 drugs are mostly preferred for ongoing systemic inflammation. Anti-IL-6 agents are very efficient in patients with a polyarticular course.


Phylogenetic diversity of a bacterial community determined from Siberian tundra soil DNA.
Genomic DNA was isolated from the active layer of tundra soil collected from the Kolyma lowland, Northeast Eurasia, near the Arctic Ocean coast. The SSU (small subunit) rRNA genes were amplified with eubacterial primers from the bulk genomic community DNA and cloned into plasmid vectors. Forty-three SSU rDNA clones were obtained, and all of them had different RFLP patterns. Phylogenetic analysis based on partial sequences (about 300 bp) established with the maximum likelihood method revealed the presence of three major and several minor groups that fell into 11 of the established lines of bacteria, and one sequence that could not be assigned to any of the described groups. Most of the clones belonged to the alpha (20.9%) and delta (25.6%) subdivisions of the Proteobacteria, with lesser proportions in the beta (9.3%) and gamma (4.7%) subdivisions, groups typically isolated from soil by culture methods. Fewer than 12% of the clones belonged to Gram-positive bacteria, and 16% of the clones were related to Fibrobacter. The majority of the clones (70%) had sequences that were 5-15% different from those in the current databases, and 7% of the clones had sequences that differed by more than 20% from those in the database. The results suggest that these tundra-derived clones are very diverse in phylogeny, and that many probably reflect new genera or families. Hence, most of the tundra soil bacterial community has never been isolated and thus the physiology and function of its dominant members appears to be unknown.


Estimation of Blood Sirolimus Concentration Based on Tacrolimus Concentration/Dose Normalized by Body Weight Ratio in Lung Transplant Patients.
Sirolimus and tacrolimus require accurate drug dosing based on their target blood levels to produce better clinical outcomes, specifically, the avoidance of drug-induced adverse effects and the maintenance of efficacy. However, because the ideal dose of sirolimus and the schedule for measuring its blood levels are unclear in lung transplant patients, an index is required for estimating sirolimus blood concentrations. The aim of this work is to study the correlation between the trough concentration/dose normalized by body weight (C0/D) ratios of sirolimus and tacrolimus in lung transplant patients. Thirteen lymphangiomyomatosis patients who underwent lung transplantation and were treated with sirolimus and tacrolimus from February 2015 to July 2018 were divided into 2 groups, one receiving twice-daily (TD, n = 6) and the other once-daily (OD, n = 7) tacrolimus formulations. The correlation between the C0/D ratio of sirolimus and patient background was evaluated using Spearman's rank correlation coefficient. Correlations between sirolimus and tacrolimus C0/D ratios or doses were analyzed by single regression analysis. Significant correlations were found between the C0/D ratios of sirolimus and tacrolimus. The regression equations from the initial data of TD and OD groups at steady state were y = 1.880x + 32.636 (adjusted R = 0.743, P = 0.017) and y = 1.684x + 38.816 (adjusted R = 0.919, P < 0.001), respectively. In addition, the regression equations from all data of TD and OD groups were y = 1.883x + 4.170 (adjusted R = 0.546, P < 0.001) and y = 1.950x + 43.188 (adjusted R = 0.898, P < 0.001), respectively. A significant correlation between the dosage of sirolimus and tacrolimus was observed only in the OD group, with relatively low accuracy. Blood sirolimus concentrations can be estimated using the C0/D ratio of tacrolimus, suggesting that the C0/D ratio of tacrolimus is an index of required sirolimus dosage and the frequency of blood sirolimus concentration measurements.


Unmet expectations of medications and care providers among patients with heart failure assessed to be poorly adherent: results from the Chronic Heart Failure Intervention to Improve MEdication Adherence (CHIME) study.
Ineffective medication management contributes to repeated hospitalisation and death among patients with heart failure. The meaning ascribed to medications and the influence of meaning on how patients manage medications is unknown. The purpose of this study was to explore the meaning and expectations associated with medication use in high-risk, non-adherent patients with heart failure. Patients ( n=265) with heart failure were screened for adherence to prescribed medication using the Morisky medication adherence scale (MMAS). Patients (MMAS score <6; n=44) participated in semistructured interviews, analysed using qualitative content analysis. Of 17 initial themes (223 representative segments), the overarching theme 'unmet expectations' consisted of two subthemes 'working to be heard' by professionals and 'resignation' to both the illness and medications. Patients' expectations were challenged by unexpected work to communicate with providers in general (72 representative segments), and specifically regarding medications (118 representative segments) and feelings of resignation regarding the medication regimen (33 representative segments). These findings suggest that unmet expectations contribute to poor medication management. Improved listening and communication by providers, to establish a common understanding and plan for managing medications may strengthen patient beliefs, resolve feelings of resignation and improve patients' ability to manage medications effectively.


The weaker sex? Vulnerable men and women's resilience to socio-economic disadvantage.
Sex differences in mortality vary over time and place as a function of social, health, and medical circumstances. The magnitude of these variations, and their response to large socioeconomic changes, suggest that biological differences cannot fully account for sex differences in survival. Drawing on a wide swath of mortality data across countries and over time, we develop a set of empiric observations with which any theory about excess male mortality and its correlates will have to contend. We show that as societies develop, M/F survival first declines and then increases, a "sex difference in mortality transition" embedded within the demographic and epidemiologic transitions. After the onset of this transition, cross-sectional variation in excess male mortality exhibits a consistent pattern of greater female resilience to mortality under socio-economic adversity. The causal mechanisms underlying these associations merit further research.


Importance of endogenous angiotensin II in the cardiovascular responses to sympathetic stimulation in conscious rabbits.
Pharmacological evidence indicates that angiotensin (Ang II) converting enzyme inhibitors attenuate cardiovascular responses to sympathetic stimulation. To investigate the physiological significance of this attenuation, the pressor and heart rate responses to bilateral carotid occlusion (BCO) were studied before and after administration of captopril and again during Ang II replacement in conscious, aortic nerve-sectioned rabbits with chronically implanted carotid occluders. In the control period, BCO produced increases (p less than 0.05) in mean arterial pressure (MAP) and heart rate (HR) of 37.3 +/- 3.0 mm Hg and 21.7 +/- 5.4 beats/min from baseline values of 79.1 +/- 2.5 mm Hg and 255.4 +/- 16.7 beats/min. Captopril (5 mg/kg i.v.) markedly reduced (p less than 0.05) both the pressor (10.2 +/- 2.6 mm Hg) and HR (5.0 +/- 4.0 beats/min) responses to BCO, in parallel with a decrease in plasma Ang II of 75%. Infusion of a subpressor dose of Ang II (5-25 ng/kg/min i.v.) increased plasma Ang II to precaptopril levels and fully restored (p less than 0.05) the pressor (33.0 +/- 5.7 mm Hg) and HR (19.8 +/- 7.7 beats/min) responses to BCO. In two additional series of experiments, the mechanism of the effects of captopril and Ang II were investigated. In the first series, cardiac baroreflex curves (pulse interval versus MAP) were generated by increasing or decreasing blood pressure with phenylephrine or nitroprusside (5-20 micrograms/kg/min i.v.). The slope of the linear region of the curve (2.9 msec/mm Hg) was not changed significantly by captopril treatment (3.1 msec/mm Hg) or Ang II replacement (3.2 msec/mm Hg), indicating that cardiac baroreflex sensitivity was not altered by blockade of the renin-angiotensin system. In the second series, the effect of captopril on the pressor response to exogenous norepinephrine (0.1-2.5 micrograms/kg/min i.v.) was tested. The response was reduced by less than 40%, indicating only a modest postsynaptic component to the action of captopril. These results provide physiological evidence for an important action of endogenous Ang II in facilitating the cardiovascular responses to sympathetic stimulation in conscious rabbits. This facilitation is not due to an action upon the baroreflex per se but results, at least in part, from a presynaptic action of Ang II.


Administration of vitamin D3 improves antimetastatic efficacy of cancer vaccine therapy of Lewis lung carcinoma.
To analyze antitumor efficacy of experimental cancer vaccine therapy combined with introduction of vitamin D3 (VD3) for treatment of Lewis lung carcinoma (3LL). Cancer vaccines composed from recombinant murine beta-defensin-2 (mBD-2) and 3LL cell lysate, or DNA, coding for mBD-2-Muc1 fusion construct cloned in pcDNA3+ vector, were prepared and used for intradermal vaccination. Experimental cancer vaccines introduced i. d. at therapeutic and prophylactic regimens to 3LL-bearing C57Bl mice, were applied alone or in combination with VD3 (administered per os) and/or low-dose cyclophosphamide (CP, administered intraperitoneal). Efficacy of treatments was analyzed by primary tumor growth dynamics indexes and by metastasis rate in vaccinated animals. As it has been shown, administration of the protein-based vaccine composed from mBD-2 and 3LL cell lysate in combination with VD3 and CP, but not in VD3 free setting, led to significant suppression of primary tumor growth (p < 0.005) and had significant antimetastatic effect. Introduction of VD3 with or without CP in the scheme of treatment with mBD- 2-Muc1-DNA vaccine at therapeutic regimen has led to significant suppression of primary tumor (p < 0.05) and metastasis volumes (p < 0.005), while in the groups of animals treated with DNA-vaccine + VD3 with or without CP at prophylactic regimen, significant antimetastatic effect (p < 0.05) and elevation of average life-span (p < 0.05) have been registered. The results of this pilot study have shown promising clinical effects of VD3 administration in combination with cancer vaccinotherapy in vivo.


The importance of psychological assessment in chronic pain.
Much confusion has surrounded the purpose of the psychological assessment in the context of chronic pain. For many clinicians, the psychological assessment is used to rule out psychiatric illness and to identify the nonmedical causes for pain and disability. In essence, it is used to identify the causes of pain that fall outside of the biomedical model. Supported by over 30 years of evidence, the bio-psycho-social model acknowledges that psychosocial factors are inherent in chronic pain and require assessment if meaningful diagnostics and treatments are to occur. Five broad categories of psychosocial assessment are relevant to chronic pain. These categories have been shown to enhance the diagnosis of the underlying forms of pain, predict the transition from acute to chronic status, and help to phenotype individuals for the discovery of the underlying mechanisms responsible for pain. Informed assessment of chronic pain needs to include relevant biological, psychological, and social domains. This article describes those domains and offers suggestions of specific instruments to use in clinical or research settings.


Pathophysiological features of asthma develop in parallel in house dust mite-exposed neonatal mice.
Asthma frequently commences in early life during airway and immune development and exposure to new environmental challenges. Endobronchial biopsies from children with asthma are abnormal, and lung function is maximally reduced by 6 years of age. As longitudinal biopsy studies are unethical in children, the relationship between development of pathology and reduced lung function is unknown. We aimed to establish a novel neonatal mouse model of allergic airways disease to investigate the developmental sequence of the pathophysiologic features of asthma. Neonatal Balb/c mice were challenged three times weekly from Day 3 of life using intranasal house dust mite (HDM) or saline for up to 12 weeks. Weekly assessments of airway inflammation and remodeling were made. Airway hyperresponsiveness (AHR) to methacholine was assessed from Week 2 onward. Total and eosinophilic inflammation was significantly increased in the lungs of HDM-exposed neonates from Week 2 onwards, and a peak was seen at 3 weeks. Goblet cells and peribronchiolar reticulin deposition were significantly increased in HDM-exposed neonates from Week 3, and peribronchiolar collagen was significantly greater from Week 4. HDM-exposed neonates had increased AHR from Week 2 onward. Although inflammation and AHR had subsided after 4 weeks without allergen challenge, the increased reticulin and collagen deposition persisted in HDM-exposed mice. Neonatal mice exposed to intranasal HDM developed eosinophilic inflammation, airway remodeling, and AHR as reported in pediatric asthma. Importantly, all abnormalities developed in parallel, not sequentially, between 2 and 3 weeks of age.


The prophylactic effect of a Eugenia aquea extract against oxidative stress and inflammation associated with the development of arthritis in an adjuvant-induced arthritis rat model.
Rheumatoid arthritis (RA) is the most common human autoimmune disease. A petroleum ether extract of Eugenia aquea (E. aquea) was analyzed by GC/MS. Antioxidant and anti-inflammatory activities were investigated in rats with adjuvant-induced arthritis (AIA). An AIA rat model received orally/daily a vehicle, diclofenac (100 mg per kg b.w.), and E. aquea extract (50 or 100 or 200 mg per kg b.w.; for 21 days). Fifty-five out of 70 compounds (97.77%) were identified: eucalyptol (34.14%), α-pinene (15.91%), l-verbenone (8.01%), camphor (7.38%) and borneol (6.74%). In an acute oral toxicity study, the E. aquea extract did not show any toxic effects in rats at 2000 mg/ kg-1. Only a high dose of the E. aquea extract or diclofenac significantly alleviated (P < 0.05-0.001) all complications observed in arthritic rats, including body weight loss, articular/extra-articular oxidative injury and synovial joint inflammation by increasing food intake as well as improving the antioxidant defense system and inflammatory marker. The dose-dependent modulatory activity of the E. aquea extract was statistically significant. It was equivalent to and sometimes even better than that of diclofenac. The present study proved the antioxidant and anti-inflammatory activities of the E. aquea extract, which could be attributed to the presence of eucalyptol and α-pinene.


Mesenchymal stem cells derived exosomal miR-323-3p promotes proliferation and inhibits apoptosis of cumulus cells in polycystic ovary syndrome (PCOS).
Polycystic ovary syndrome (PCOS) is a heterogeneous reproductive disease. Adipose mesenchymal stem cells (AMSCs) can produce a mass of exosomes. The objective of this study was to determine the effects of exosomal miR-323-3p on cumulus cells (CCs) of PCOS patients. Exosomal miR-323-3p were collected from modified AMSCs. Real-time PCR, western blots, MTT assays, flow cytometry, luciferase reporter assays and a letrozole-induced PCOS mouse model were used to identify mechanisms of exosomal miR-323-3p on CCs. The results revealed that miR-323-3p expression was upregulated in AMSCs, exosomes and CCs. Upregulated miR-323-3p promoted cell proliferation and suppressed apoptosis in CCs, while miR-323-3p inhibitor exerted opposite roles in exosome-treated CCs. Moreover, PDCD4 was upregulated in PCOS CCs, displayed an inverse expression pattern to those of miR-323-3p, and was a direct target of miR-323-3p. Overexpression of PDCD4 reversed the effects of upregulated miR-323-3p on CCs. Serum FSH, LH and testosterone were upregulated while E2 levels were downregulated in the PCOS mice. Upregulation of miR-323-3p alleviated PCOS by suppressing CCs' apoptosis through targeting PDCD4 in vivo. The results demonstrated that exosomal miR-323-3p promoted cell proliferation and inhibited apoptosis in CCs through targeting PDCD4 in PCOS. This study provides insight into developing new therapeutic strategies for PCOS.


Additive manufacturing of reconstruction devices for maxillofacial surgery: design and accuracy assessment of a mandibular plate prototype.
Additive manufacturing (AM) presents unique opportunities for medical applications and in particular in maxillofacial surgery for developing patient specific implants. The quality assessment of additive manufactured products is an essential aspect for the real introduction in health services. In this framework, the purpose of the present study is to investigate the possibility of developing prototypes of mandibular plates as preoperative surgical planning models, by verification of design, analysis of internal structure integrity and evaluation of the effects of variables involved in AM processes. A PolyJet three-dimensional (3D) printing system is used in the study due to its very fine resolution.The computer aided design (CAD) models of the implants were converted to stereolithography (STL) file formats in different STL conversion resolutions and then printed using commercial prototyping polymers to observe the effect of model resolution. Finite element analysis (FEA) was conducted to study the capability of the designed mandibular plate to support the involved biomechanical loads. Micro-computed tomography (micro-CT) analysis was performed to verify the dimensions and the internal defects of the printed objects, considering that the presence of defects can affect the quality and compromise the final performance. Results were analyzed to understand the effect of the 3D printing process flow conditions on the obtained prototypes. Relative error in reference to the CAD models mainly evidenced the difference in resolution due to STL files and the effect of the design. No anomalies and defects were detected inside the evaluated samples.


A G1-like state allows HIV-1 to bypass SAMHD1 restriction in macrophages.
An unresolved question is how HIV-1 achieves efficient replication in terminally differentiated macrophages despite the restriction factor SAMHD1. We reveal inducible changes in expression of cell cycle-associated proteins including MCM2 and cyclins A, E, D1/D3 in macrophages, without evidence for DNA synthesis or mitosis. These changes are induced by activation of the Raf/MEK/ERK kinase cascade, culminating in upregulation of CDK1 with subsequent SAMHD1 T592 phosphorylation and deactivation of its antiviral activity. HIV infection is limited to these G1-like phase macrophages at the single-cell level. Depletion of SAMHD1 in macrophages decouples the association between infection and expression of cell cycle-associated proteins, with terminally differentiated macrophages becoming highly susceptible to HIV-1. We observe both embryo-derived and monocyte-derived tissue-resident macrophages in a G1-like phase at frequencies approaching 20%, suggesting how macrophages sustain HIV-1 replication in vivo Finally, we reveal a SAMHD1-dependent antiretroviral activity of histone deacetylase inhibitors acting via p53 activation. These data provide a basis for host-directed therapeutic approaches aimed at limiting HIV-1 burden in macrophages that may contribute to curative interventions.


Pharmacokinetics and pharmacodynamics of lumefantrine (benflumetol) in acute falciparum malaria.
The objective of this study was to conduct a prospective population pharmacokinetic and pharmacodynamic evaluation of lumefantrine during blinded comparisons of artemether-lumefantrine treatment regimens in uncomplicated multidrug-resistant falciparum malaria. Three combination regimens containing an average adult lumefantrine dose of 1,920 mg over 3 days (four doses) (regimen A) or 2,780 mg over 3 or 5 days (six doses) (regimen B or C, respectively) were given to 266 Thai patients. Detailed observations were obtained for 51 hospitalized adults, and sparse data were collected for 215 patients of all ages in a community setting. The population absorption half-life of lumefantrine was 4.5 h. The model-based median (5th and 95th percentiles) peak plasma lumefantrine concentrations were 6.2 (0.25 and 14.8) microgram/ml after regimen A, 9. 0 (1.1 and 19.8) microgram/ml after regimen B, and 8 (1.4 and 17.4) microgram/ml after regimen C. During acute malaria, there was marked variability in the fraction of drug absorbed by patients (coefficient of variation, 150%). The fraction increased considerably and variability fell with clinical recovery, largely because food intake was resumed; taking a normal meal close to drug administration increased oral bioavailability by 108% (90% confidence interval, 64 to 164) (P, 0.0001). The higher-dose regimens (B and C) gave 60 and 100% higher areas under the concentration-time curves (AUC), respectively, and thus longer durations for which plasma lumefantrine concentrations exceeded the putative in vivo MIC of 280 microgram/ml (median for regimen B, 252 h; that for regimen C, 298 h; that for regimen A, 204 h [P, 0.0001]) and higher cure rates. Lumefantrine oral bioavailability is very dependent on food and is consequently poor in acute malaria but improves markedly with recovery. The high cure rates with the two six-dose regimens resulted from increased AUC and increased time at which lumefantrine concentrations were above the in vivo MIC.


Variability of distribution of Ca(2+)/calmodulin-dependent kinase II at mixed synapses on the mauthner cell: colocalization and association with connexin 35.
In contrast to chemical transmission, few proteins have been shown associated with gap junction-mediated electrical synapses. Mixed (electrical and glutamatergic) synaptic terminals on the teleost Mauthner cell known as "Club endings" constitute because of their unusual large size and presence of connexin 35 (Cx35), an ortholog of the widespread mammalian Cx36, a valuable model for the study of electrical transmission. Remarkably, both components of their mixed synaptic response undergo activity-dependent potentiation. Changes in electrical transmission result from interactions with colocalized glutamatergic synapses, the activity of which leads to the activation of Ca(2+)/calmodulin-dependent kinase II (CaMKII), required for the induction of changes in both forms of transmission. However, the distribution of this kinase and potential localization to electrical synapses remains undetermined. Taking advantage of the unparalleled experimental accessibility of Club endings, we explored the presence and intraterminal distribution of CaMKII within these terminals. Here we show that (1) unlike other proteins, both CaMKII labeling and distribution were highly variable between contiguous contacts, and (2) CaMKII was not restricted to the periphery of the terminals, in which glutamatergic synapses are located, but also was present at the center in which gap junctions predominate. Accordingly, double immunolabeling indicated that Cx35 and CaMKII were colocalized, and biochemical analysis showed that these proteins associate. Because CaMKII characteristically undergoes activity-dependent translocation, the observed variability of labeling likely reflects physiological differences between electrical synapses of contiguous Club endings, which remarkably coexist with differing degrees of conductance. Together, our results indicate that CaMKII should be considered a component of electrical synapses, although its association is nonobligatory and likely driven by activity.


Removal of carbamazepine and sulfamethoxazole by MBR under anoxic and aerobic conditions.
This study reveals for the first time that near-anoxic conditions (dissolved oxygen, DO=0.5 mg/L) can be a favorable operating regime for the removal of the persistent micropollutant carbamazepine by MBR treatment. The removal efficiencies of carbamazepine and sulfamethoxazole by an MBR were systematically examined and compared under near-anoxic (DO=0.5 mg/L) and aerobic (DO>2 mg/L) conditions. Preliminary batch tests confirmed that sulfamethoxazole is amenable to both aerobic and anoxic biotransformation. However, carbamazepine-a known persistent compound-showed degradation only under an anoxic environment. In good agreement with the batch tests, during near-anoxic operation, under a high loading of 750 μg/Ld, an exceptionally high removal (68±10%) of carbamazepine was achieved. In contrast, low removal efficiency (12±11%) of carbamazepine was observed during operation under aerobic conditions. On the other hand, an average removal efficiency of 65% of sulfamethoxazole was achieved irrespective of the DO concentrations.


Creutzfeldt-Jakob Disease-Like Periodic Sharp Wave Complexes in Voltage-Gated Potassium Channel-Complex Antibodies Encephalitis: A Case Report.
Voltage-gated potassium channel-complex antibodies (VGKC-cAbs) encephalitis, a treatable autoantibody encephalopathy, has been previously reported to clinically mimic sporadic Creutzfeldt-Jakob disease. Among available clinical clues to distinguish them, periodic sharp wave complexes, a typical finding in sporadic Creutzfeldt-Jakob disease, have never been reported in association with VGKC-cAbs encephalitis. A 76-year-old man was transferred to a tertiary neurology center with a clinical history of 6-month weight loss, cognitive disturbance, and nonspecific generalized weakness. He had two seizures the month before transfer and then evolved to severe encephalopathy, requiring mechanical ventilation. Periodic sharp wave complexes every 1 to 2 seconds over slowed background were found on EEG, and MRI showed cerebellar and bifrontal cortical T2/FLAIR/DWI hypersignal without restricted diffusion on ADC mapping. Pancorporal positron emission tomography scan was negative. An immunotherapy trial did not improve the patient condition. Therefore, he died after life support withdrawal. Brain autopsy revealed mononuclear neocortex infiltrate without significant spongiosis, and the anti-VGKC test showed a seropositivity of 336 pmol/L (normal, 0-31), 3 month after the patient deceased. This is the first reported case of VGKC-cAbs encephalitis associated with periodic sharp wave complexes on EEG, which further confuse the differential diagnosis with sporadic Creutzfeldt-Jakob disease. However, the cortical DWI hypersignal without restriction seems to remain a way to discriminate these two entities appropriately, when present. These clues are of paramount importance because VGKC-cAbs encephalitis is a treatable disease.


HLA Associations in a Cohort of Children With Juvenile Idiopathic Arthritis With and Without Uveitis.
Juvenile idiopathic arthritis (JIA)-associated uveitis can lead to ocular complications and vision loss. Alleles HLA-DRB1*08, *11, and *13 are risk alleles for JIA, whereas HLA-DRB1*11 and *13 alleles increase uveitis susceptibility. We examined the association of common HLA-DRB1 alleles in children with JIA alone and JIA-associated uveitis. High-resolution HLA-DRB1 genotyping was performed in 107 children with oligoarticular and polyarticular rheumatoid factor (RF) negative JIA and 373 non-Hispanic white controls. Children with JIA alone and JIA-associated uveitis were of similar race, ethnicity, sex, and age at arthritis diagnosis. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. There were 47 children with JIA-associated uveitis and 60 with JIA alone. Compared to controls, only children with JIA-associated uveitis had increased odds of carriage of HLA-DRB1*11 (OR, 2.2 95% [CI, 1.1-4.3], P = 0.023). There also was increased carriage of HLA-DRB1*08 and *13 (OR, 12.6 [95% CI, 2.0-77.8], P = 0.011). Compared to controls and children with JIA alone, those with JIA-associated uveitis had increased odds of carriage of HLA-DRB1*11 and *13 (OR, 9 [95% CI, 2.8-29.0], P < 0.0001 and OR, 8.6 [95% CI, 1.0-74.4], P = 0.042), respectively. We report the novel finding that carriage of HLA-DRB1*11 and *13 appears to increase the risk of uveitis in children with JIA.


Coil embolization of a fistula from the right inferior phrenic artery to the right pulmonary artery with involvement of further arteries: A rare case report.
A 51-year-old female patient was presenting dyspnea for more than a year with no previous lung infections or surgery. Initially, a diagnostic computed tomography was made, showing a rare arterio-arterial malformation between the right inferior phrenic and right pulmonary artery leading into a vascular bundle in the middle lung lobe. Due to the patients' dyspnea and massive extent of malformation, the indication for transcatheter arterial embolization was made. The first transcatheter arterial embolization procedure involved the inferior phrenic and a selective branch of the internal thoracic artery. Interventional angiography as well as computed tomography revealed further extend of the malformation showing a connection of right lateral thoracic, hepatic, and inferior epigastric artery to the fistula. After one month, a second transcatheter arterial embolization of these arteries as well as a second approach of the proximal internal thoracic artery was performed. In the follow-up the patient described a substantial improvement of her dyspnea and showed no signs of infections. A phrenic artery to pulmonary artery fistula is an extremely rare case occurring congenital or acquired. Patients may be asymptomatic or present, among others, dyspnea, hemoptysis, pulmonary infections and congestive heart failure. Symptomatic patients require treatment using transcatheter arterial embolization or surgical resection. The patient had dyspnea and a substantial extent of malformation with possibly complicated clinical course. The recommended less invasive treatment using transcatheter arterial embolization was successfully performed. In conclusion, our patient represented a rare congenital case of systemic and pulmonary artery communication, which we were able to treat sufficiently with coil embolization.


Adverse outcome pathways: a conceptual framework to support ecotoxicology research and risk assessment.
Ecological risk assessors face increasing demands to assess more chemicals, with greater speed and accuracy, and to do so using fewer resources and experimental animals. New approaches in biological and computational sciences may be able to generate mechanistic information that could help in meeting these challenges. However, to use mechanistic data to support chemical assessments, there is a need for effective translation of this information into endpoints meaningful to ecological risk-effects on survival, development, and reproduction in individual organisms and, by extension, impacts on populations. Here we discuss a framework designed for this purpose, the adverse outcome pathway (AOP). An AOP is a conceptual construct that portrays existing knowledge concerning the linkage between a direct molecular initiating event and an adverse outcome at a biological level of organization relevant to risk assessment. The practical utility of AOPs for ecological risk assessment of chemicals is illustrated using five case examples. The examples demonstrate how the AOP concept can focus toxicity testing in terms of species and endpoint selection, enhance across-chemical extrapolation, and support prediction of mixture effects. The examples also show how AOPs facilitate use of molecular or biochemical endpoints (sometimes referred to as biomarkers) for forecasting chemical impacts on individuals and populations. In the concluding sections of the paper, we discuss how AOPs can help to guide research that supports chemical risk assessments and advocate for the incorporation of this approach into a broader systems biology framework.


Modification of agricultural waste/by-products for enhanced phosphate removal and recovery: potential and obstacles.
There is a growing trend to employ agricultural waste/by-products (AWBs) as the substrates for the development of phosphate biosorbents. Nevertheless, due to the lack of anion binding sites, natural AWBs are usually inefficient in phosphate decontamination. Consequently, modification plays a vital role in improving phosphate sorption's property of raw AWBs. This review paper evaluates all existing methods of modification. The literatures indicate that modification can significantly improve phosphate removal ability of AWBs by retaining phosphate ion onto modified AWBs principally via ion exchange (electrostatic interaction) and ligand exchange mechanisms. So far, little work has been done on the beneficial use of modified AWBs for the phosphorus recovery from aqueous solutions. The poor recyclability of modified AWBs could be responsible for their limited application. Hence, further study is essential to search for novel, cost-effective, and green methods of modification.


Genes that fight infection: what the Drosophila genome says about animal immunity.
From deciphering the principles of heredity to identifying the genes that control development, the fruit fly Drosophila melanogaster is being used to deconstruct an increasing number of biological processes. Genetic studies of Drosophila responses to microbial infection have identified regulators of innate immunity that are functionally conserved in mammals. These recent findings highlight the ancient origins of animal immune responses and demonstrate the potential of Drosophila for dissecting host-pathogen interactions. The sequencing of the Drosophila genome both enhances genetic approaches and provides new clues for the identification of key components of innate immunity. This article summarizes how information gained from genomic analysis contributes to our understanding of how animals cope with infectious disease.


Patient perspectives on the linkage of health data for research: Insights from an online patient community questionnaire.
To examine the patient perspective on the risks and benefits of linking existing data sources for research. Between December 2015 and February 2016, we fielded a questionnaire in PatientsLikeMe, an online patient community representing over 2500 health conditions. The questionnaire was developed using subject matter expertise and patient feedback from a concept elicitation phase (N = 57 patients). The final questionnaire consisted of 37 items. Of n = 5741 who opened the email invitation, n = 3516 respondents completed the questionnaire (61.2%). Of these, 73.8% were women, 86.4% were Caucasian, 14.5% were 65 or older, and 44.9% had completed college or post-graduate education. Questionnaire respondents indicated that the most important benefits of sharing data were "helping my doctor make better decisions about my health" (94%) and "helping make new therapies available faster" (94%). The most important data sharing risk identified was health data being "stolen by hackers" (87%). Of 693 patients who were not comfortable with researchers accessing their de-identified data, most reported that their comfort levels would increase if they were able to learn how their data was protected (84%). In general, responders felt more comfortable when unique identifiers such as social security number (90%) and insurance ID (82%) were removed from the data for linkage and research use. The majority of patients in a US-based online community are comfortable with researchers accessing their de-identified data for research purposes. Developing methods to link databases minimizing the exposure of unique identifiers may improve patient comfort levels with linking data for research purposes.


Mechanism of Shiga Toxin Clustering on Membranes.
The bacterial Shiga toxin interacts with its cellular receptor, the glycosphingolipid globotriaosylceramide (Gb3 or CD77), as a first step to entering target cells. Previous studies have shown that toxin molecules cluster on the plasma membrane, despite the apparent lack of direct interactions between them. The precise mechanism by which this clustering occurs remains poorly defined. Here, we used vesicle and cell systems and computer simulations to show that line tension due to curvature, height, or compositional mismatch, and lipid or solvent depletion cannot drive the clustering of Shiga toxin molecules. By contrast, in coarse-grained computer simulations, a correlation was found between clustering and toxin nanoparticle-driven suppression of membrane fluctuations, and experimentally we observed that clustering required the toxin molecules to be tightly bound to the membrane surface. The most likely interpretation of these findings is that a membrane fluctuation-induced force generates an effective attraction between toxin molecules. Such force would be of similar strength to the electrostatic force at separations around 1 nm, remain strong at distances up to the size of toxin molecules (several nanometers), and persist even beyond. This force is predicted to operate between manufactured nanoparticles providing they are sufficiently rigid and tightly bound to the plasma membrane, thereby suggesting a route for the targeting of nanoparticles to cells for biomedical applications.


Positive and negative symptom scores are correlated with activation in different brain regions during facial emotion perception in schizophrenia patients: a voxel-based sLORETA source activity study.
Schizophrenia is one of the most devastating of all mental illnesses, and has dimensional characteristics that include both positive and negative symptoms. One problem reported in schizophrenia patients is that they tend to show deficits in face emotion processing, on which negative symptoms are thought to have stronger influence. In this study, four event-related potential (ERP) components (P100, N170, N250, and P300) and their source activities were analyzed using EEG data acquired from 23 schizophrenia patients while they were presented with facial emotion picture stimuli. Correlations between positive and negative syndrome scale (PANSS) scores and source activations during facial emotion processing were calculated to identify the brain areas affected by symptom scores. Our analysis demonstrates that PANSS positive scores are negatively correlated with major areas of the left temporal lobule for early ERP components (P100, N170) and with the right middle frontal lobule for a later component (N250), which indicates that positive symptoms affect both early face processing and facial emotion processing. On the other hand, PANSS negative scores are negatively correlated with several clustered regions, including the left fusiform gyrus (at P100), most of which are not overlapped with regions showing correlations with PANSS positive scores. Our results suggest that positive and negative symptoms affect independent brain regions during facial emotion processing, which may help to explain the heterogeneous characteristics of schizophrenia.


The effects of work intensity on adolescent mental health, achievement, and behavioral adjustment: new evidence from a prospective study.
This article examines the effects of work intensity on adolescent mental health, academic achievement, and behavioral adjustment. Questionnaire data were collected yearly from an initial panel of 1,000 randomly selected ninth graders (14-15 years old). Consistent with other studies, students who worked at higher intensity engaged in more alcohol use. The methodological strengths of this research (a representative panel studied prospectively over a 4-year period with minimal attrition and an analysis incorporating key control and lagged variables) provide strong evidence that adolescent work fosters alcohol use. The contention that work of high intensity has deleterious effects on mental health, academic achievement, and 2 other indicators of behavioral adjustment did not withstand our stringent tests. However, high school seniors who worked at moderate intensity (1-20 hours per week) had higher grades than both nonworkers and students who worked more hours per week.


"TEG talk": expanding clinical roles for thromboelastography and rotational thromboelastometry.
Viscoelastic assays (VEAs) that include thromboelastography and rotational thromboelastometry add value to the investigation of coagulopathies and goal-directed management of bleeding by providing a complete picture of clot formation, strength, and lysis in whole blood that includes the contribution of platelets, fibrinogen, and coagulation factors. Conventional coagulation assays have several limitations, such as their lack of correlation with bleeding and hypercoagulability; their inability to reflect the contribution of platelets, factor XIII, and plasmin during clot formation and lysis; and their slow turnaround times. VEA-guided transfusion algorithms may reduce allogeneic blood exposure during and after cardiac surgery and in the emergency management of trauma-induced coagulopathy and hemorrhage. However, the popularity of VEAs for other indications is driven largely by extrapolation of evidence from cardiac surgery, by the drawbacks of conventional coagulation assays, and by institution-specific preferences. Robust diagnostic studies validating and standardizing diagnostic cutoffs for VEA parameters and randomized trials comparing VEA-guided algorithms with standard care on clinical outcomes are urgently needed. Lack of such studies represents the biggest barrier to defining the role and impact of VEA in clinical care.


Assessment and treatment of mood disorders in the context of substance abuse.
Recognition and management of mood symptoms in individuals using alcohol and/or other drugs represent a daily challenge for clinicians in both inpatient and outpatient treatment settings. Diagnosis of underlying mood disorders in the context of ongoing substance abuse requires careful collection of psychiatric history, and is often critical for optimal treatment planning and outcomes. Failure to recognize major depression or bipolar disorders in these patients can result in increased relapse rates, recurrence of mood episodes, and elevated risk of completed suicide. Over the past decade, epidemiologic research has clarified the prevalence of comorbid mood disorders in substance-dependent individuals, overturning previous assumptions that depression in these patients is simply an artifact of intoxication and/or withdrawal, therefore requiring no treatment. However, our understanding of the bidirectional relationships between mood and substance use disorders in terms of their course(s) of illness and prognoses remains limited. Like-wise, strikingly little treatment research exists to guide clinical decision making in co-occurring mood and substance use disorders, given their high prevalence and public health burden. Here we overview what is known and the salient gaps of knowledge where data might enhance diagnosis and treatment of these complicated patients.


Aortic propagation velocity does not correlate with classical aortic stiffness parameters in healthy individuals.
Aortic stiffness is an important cardiovascular risk marker, which can be determined using different noninvasive techniques. Aortic propagation velocity (APV) has recently been established as a novel echocardiographic parameter of aortic stiffness. This study aimed to investigate the association between APV and the classical echocardiography-derived aortic stiffness parameters, aortic distensibility (AD) and aortic strain (AS), in a group of otherwise healthy individuals. In total, 97 consecutive healthy subjects were recruited in this observational study. APV was measured using color M-mode echocardiography from the suprasternal window in the descending aorta. AS and AD were calculated using clinical blood pressure and the M-mode echocardiography-derived aortic diameters. Correlation analyses were performed between cardiovascular risk factors related to increased aortic stiffness (age, obesity, and blood pressure) and measured stiffness parameters (APV, AS, and AD). Correlation analyses were also performed among the measured stiffness parameters. Good correlation of age, blood pressure, and BMI with AS and AD was observed. One-on-one correlation of age, blood pressure, and BMI with APV was not observed. No correlation was observed between APV and AS (r=-0.05, p=0.6) or between APV and AD (r=-0.17, p=0.8). Although APV has been proposed as a novel and practical echocardiographic parameter of aortic stiffness, especially in patients with coronary artery disease, correlations between classical stiffness parameters (AS and AD) and APV were absent in healthy individuals at low-intermediate risk. The clinical and research applicability of APV should be further evaluated.


Visualizing picometric quantum ripples of ultrafast wave-packet interference.
Interference fringes in vibrating molecules are a signature of quantum mechanics, but are often so short-lived and closely spaced that they elude visualization. We have experimentally visualized dynamical quantum interferences, which appear and disappear in less than 100 femtoseconds in the iodine molecule synchronously with the periodic crossing of two counterpropagating nuclear wave packets. The obtained images have picometer and femtosecond spatiotemporal resolution, representing a detailed picture of the quantum interference.


Purification, characterization and mass spectrometric sequencing of a thermophilic glucuronoyl esterase from Sporotrichum thermophile.
The cellulolytic system of the thermophilic fungus Sporotrichum thermophile contains a recently discovered esterase that may hydrolyze the ester linkage between the 4-O-methyl-D-glucuronic acid of glucuronoxylan and lignin alcohols. The glucuronoyl esterase named StGE1 was purified to homogeneity with a molecular mass of M(r) 58 kDa and pI 6.7. The enzyme activity was optimal at pH 6.0 and 60 degrees C. The esterase displayed a narrow pH range stability at pH 8.0 and retained 50% of its activity after 430 and 286 min at 50 and 55 degrees C, respectively. The enzyme was active on substrates containing glucuronic acid methyl ester, showing a lower catalytic efficiency on 4-nitrophenyl 2-O-(methyl-4-O-methyl-alpha-d-glucopyranosyluronate)-beta-D-xylopyranoside than its mesophilic counterparts reported in the literature, which is typical of thermophilic enzymes. StGE1 was proved to be a modular enzyme containing a noncatalytic carbohydrate-binding module. LC-MS/MS analysis provided peptide mass and sequence information that facilitated the identification and classification of StGE1 as a family 15 glucuronoyl esterase that showed the highest homology with the hypothetical glucuronoyl esterase CHGG_10774 of Chaetomium globosum CBS 148.51. This work represents the first example of the purification and identification of a thermophilic glucuronoyl esterase from S. thermophile.


Hybridization assays using an expressible DNA fragment encoding firefly luciferase as a label.
We report the use of a new label, an expressible enzyme-coding DNA fragment, for nucleic acid hybridization assays. The DNA label contains a firefly luciferase coding sequence downstream from a T7 RNA polymerase promoter. The target DNA (200 bp) is denatured and hybridized simultaneously with two oligonucleotide probes. One of the probes is immobilized in microtiter wells, via the digoxigenin/anti-digoxigenin interaction, and the other probe is biotinylated. After completion of the hybridization, the hybrids are reacted with a streptavidin-luciferase DNA complex. Subsequently, the solid-phase bound DNA is expressed by coupled transcription/ translation. The synthesized luciferase catalyzes the luminescent reaction of luciferin with O2 and ATP. The luminescence is linearly related to the amount of target DNA in the range of 5-5000 amol. The CVs obtained for 20 and 100 amol of target are 6.5% and 10.8%, respectively (n = 4).


Suppression of reflection peaks in passbands of multilayer dielectric filters.
In multilayer dielectric mirrors with one or more high reflectance bands in the near-ir and visible regions, large anomalous reflection peaks usually appear in the passbands. Their origin is traced to layer thickness inequality caused by unequal dispersions of the high and low index materials, and the common practice of monitoring layer thicknesses at stop bands. A procedure is described which takes account of the variations in materials dispersion in different coating laboratories, suppresses the anomalous reflection peaks, and locates the high reflection bands at their required wavelengths, with a relatively simple monitoring procedure.


The impact of generic reference pricing in Italy, a decade on.
The generic reference price (GRP) was introduced in Italy in 2001. The main purpose of this paper is: (a) producing evidence regarding the effect of GRP on prices; (b) testing the hypothesis that there is a reallocation of demand from the genericated (and reference-priced) molecules to patent-protected products that have the same therapeutic indication. The analysis used a unique dataset of quantities and revenues of six therapeutic groups that were observed for more than a decade. Difference-in-differences analysis is applied. Prices are adjusted for all the regulatory interventions in the ten years of observations, to control for confounding impact of these interventions. On average, prices dropped 13% more in groups to which GRP was applied than in other groups. Moreover, each entry of a new generic was associated with a price drop of around 2.8%. On the other hand, GRP did not induce any significant switching towards in-patent molecules. We provide the first empirical results of the impact of GRP on prices in Italy and evidence that GRP cannot be held solely responsible for the often reported demand reallocation towards new and in-patent molecules.


Manipulation and Mobilization for Treating Chronic Nonspecific Neck Pain: A Systematic Review and Meta-Analysis for an Appropriateness Panel.
Mobilization and manipulation therapies are widely used by patients with chronic nonspecific neck pain; however, questions remain around efficacy, dosing, and safety, as well as how these approaches compare to other therapies. Based on published trials, to determine the efficacy, effectiveness, and safety of various mobilization and manipulation therapies for treatment of chronic nonspecific neck pain. A systematic literature review and meta-analysis. We identified studies published between January 2000 and September 2017, by searching multiple electronic databases, examining reference lists, and communicating with experts. We selected randomized controlled trials comparing manipulation and/or mobilization therapies to sham, no treatment, each other, and other active therapies, or when combined as multimodal therapeutic approaches. We assessed risk of bias by using the Scottish Intercollegiate Guidelines Network criteria. When possible, we pooled data using random-effects meta-analysis. Grading of Recommendations, Assessment, Development, and Evaluation was applied to determine the confidence in effect estimates. This project was funded by the National Center for Complementary and Integrative Health under award number U19AT007912 and ultimately used to inform an appropriateness panel. A total of 47 randomized trials (47 unique trials in 53 publications) were included in the systematic review. These studies were rated as having low risk of bias and included a total of 4,460 patients with nonspecific chronic neck pain who were being treated by a practitioner using various types of manipulation and/or mobilization interventions. A total of 37 trials were categorized as unimodal approaches and involved thrust or nonthrust compared with sham, no treatment, or other active comparators. Of these, only 6 trials with similar intervention styles, comparators, and outcome measures/timepoints were pooled for meta-analysis at 1, 3, and 6 months, showing a small effect in favor of thrust plus exercise compared to an exercise regimen alone for a reduction in pain and disability. Multimodal approaches appeared to be effective at reducing pain and improving function from the 10 studies evaluated. Health-related quality of life was seldom reported. Some 22/47 studies did not report or mention adverse events. Of the 25 that did, either no or minor events occurred. The current evidence is heterogeneous, and sample sizes are generally small. Studies published since January 2000 provide low-moderate quality evidence that various types of manipulation and/or mobilization will reduce pain and improve function for chronic nonspecific neck pain compared to other interventions. It appears that multimodal approaches, in which multiple treatment approaches are integrated, might have the greatest potential impact. The studies comparing to no treatment or sham were mostly testing the effect of a single dose, which may or may not be helpful to inform practice. According to the published trials reviewed, manipulation and mobilization appear safe. However, given the low rate of serious adverse events, other types of studies with much larger sample sizes would be required to fully describe the safety of manipulation and/or mobilization for nonspecific chronic neck pain. Chronic neck pain, nonspecific, chiropractic, manipulation, mobilization, systematic review, meta-analysis, appropriateness.


Treatment of periprosthetic femoral fractures following total hip arthroplasty with femoral component revision.
Revision total hip arthroplasty is indicated for most periprosthetic fractures that occur around the stem of the femoral implant. The purpose of the present study was to assess the results and complications of revision total hip arthroplasty for the treatment of periprosthetic femoral fractures. We evaluated 118 hips in 116 patients who underwent revision total hip arthroplasty because of an acute Vancouver type-B periprosthetic femoral fracture. The femoral implant used for the revision was a cemented stem in forty-two hips, a proximally porous-coated uncemented stem in twenty-eight, an extensively porous-coated stem in thirty, and an allograft-prosthesis composite or tumor prosthesis in eighteen. The mean duration of follow-up was 5.4 years. Kaplan-Meier analysis demonstrated that the probability of survival was 90% at five years and 79.2% at ten years with revision or removal of the femoral implant for any reason as the end point. Sixteen femoral components were rerevised: ten were rerevised because of loosening; three, because of loosening in association with a fracture nonunion; two, because of recurrent dislocation; and one, because of a new periprosthetic fracture. Additionally, six femoral implants were resected because of deep infection (five) or prosthetic loosening (one). Radiographs of the ninety-six hips with a surviving implant showed that twenty-one had evidence of loosening of the femoral implant, four had a nonunion of the femoral fracture, and two had both a nonunion and loosening of the femoral implant. Revision total hip arthroplasty for the treatment of a periprosthetic fracture around the stem of the femoral implant successfully restored function for most patients. The greatest long-term problems were prosthetic loosening and fracture nonunion. Better results were seen when an uncemented, extensively porous-coated stem was used.


The Emergence of Social Norms and Conventions.
The utility of our actions frequently depends upon the beliefs and behavior of other agents. Thankfully, through experience, we learn norms and conventions that provide stable expectations for navigating our social world. Here, we review several distinct influences on their content and distribution. At the level of individuals locally interacting in dyads, success depends on rapidly adapting pre-existing norms to the local context. Hence, norms are shaped by complex cognitive processes involved in learning and social reasoning. At the population level, norms are influenced by intergenerational transmission and the structure of the social network. As human social connectivity continues to increase, understanding and predicting how these levels and time scales interact to produce new norms will be crucial for improving communities.


Reply to Chen et al.: Parametric methods for cluster inference perform worse for two-sided t-tests.
One-sided t-tests are commonly used in the neuroimaging field, but two-sided tests should be the default unless a researcher has a strong reason for using a one-sided test. Here we extend our previous work on cluster false positive rates, which used one-sided tests, to two-sided tests. Briefly, we found that parametric methods perform worse for two-sided t-tests, and that nonparametric methods perform equally well for one-sided and two-sided tests.


A bibliometric analysis of literature on malaria vector resistance: (1996 - 2015).
Emergence of insecticide resistance in malaria vectors is a real threat to future goals of elimination and control of malaria. Therefore, the objective of this study was to assess research trend on insecticide resistance of Anopheles mosquito. In specific, number of publications, countries, institutions, and authors' research profile, citation analysis, international collaborations, and impact of journals publishing documents on insecticide resistance will be presented. It was conducted via Scopus search engine which was used to retrieve relevant data. Keywords used were based on literature available on this topic. The duration of study was set from 1996-2015. A total of 616 documents, mainly as original research articles (n = 569; 92.37%) were retrieved. The average number of citations per article was 26.36. Poisson log-linear regression analysis indicated that there was a 6.00% increase in the number of publications for each extra article on pyrethroid resistance. A total of 82 different countries and 1922 authors participated in publishing retrieved articles. The United Kingdom (UK) ranked first in number of publications followed by the United States of America (USA) and France. The top ten productive countries included seven African countries. The UK had collaborations mostly with Benin (relative link strength = 46). A total of 1817 institution/ organizations participated in the publication of retrieved articles. The most active institution/ organization was Liverpool School of Tropical Medicine. Retrieved articles were published in 134 different scientific peer reviewed journals. The journal that published most on this topic was Malaria Journal (n = 101; 16.4%). Four of the top active authors were from South Africa and two were from the UK. Three of the top ten cited articles were published in Insect Molecular Biology journal. Six articles were about pyrethroid resistance and at least two were about DDT resistance. Publications on insecticide resistance in malaria vector has gained momentum in the past decade. International collaborations enhanced the knowledge about the situation of vector resistance in countries with endemic malaria. Molecular biology of insecticide resistance is the key issue in understanding and overcoming this emerging problems.


Microbial Production of Violacein and Process Optimization for Dyeing Polyamide Fabrics With Acquired Antimicrobial Properties.
In the present study, crude bacterial extract containing violacein is investigated for the preparation of antimicrobial polyamide fabrics. The optimal culture conditions of Janthinobacterium lividum (JL) for maximum biomass and violacein production were found to be 25°C, pH 7.0, while the addition of ampicillin of 0.2 mg mL-1 in the small scale increased violacein production 1.3-fold. In scale-up trials, the addition of 1% (v/v) glycerol in a fed-batch bioreactor, resulted in fivefold extracted crude violacein increase with final concentration of 1.828 g L-1. Polyamide 6.6 fabrics were dyed following three different processes; through simultaneous fermentation and dyeing (SFD), by incubating the fabric in the sonicated bacterial culture after fermentation and by using cell-free extract containing violacein. Maximum color change (ΔE) and color strength (K/S) obtained for SFD fabrics were 74.81 and 22.01, respectively, while no alteration of fastness and staining of dye at acid and alkaline perspiration or at water was indicated. The dyed fabrics presented significant antifungal activity against Candida albicans, C. parapsilosis, and C. krusei, as well as antibacterial properties against Escherichia coli, Staphylococcus aureus, and the S. aureus MRSA. We have shown that J. lividum cultures can be successfully used for violacein production and for simultaneous dying of fabrics resulting in dyed fabrics with antimicrobial properties without utilization of organic solvents.


Activity monitors for increasing physical activity in adult stroke survivors.
Stroke is the third leading cause of disability worldwide. Physical activity is important for secondary stroke prevention and for promoting functional recovery. However, people with stroke are more inactive than healthy age-matched controls. Therefore, interventions to increase activity after stroke are vital to reduce stroke-related disability. To summarise the available evidence regarding the effectiveness of commercially available, wearable activity monitors and smartphone applications for increasing physical activity levels in people with stroke. We searched the Cochrane Stroke Group Trials Register, CENTRAL, MEDLINE, Embase, CINAHL, SPORTDiscus, and the following clinical trial registers: WHO International Clinical Trials Registry Platform, Clinical Trials, EU Clinical Trial Register, ISRCTN Registry, Australian and New Zealand Clinical Trial Registry, and Stroke Trials Registry to 3 March 2018. We also searched reference lists, Web of Science forward tracking, and Google Scholar, and contacted trial authors to obtain further data if required. We did not restrict the search on language or publication status. We included all randomised controlled trials (RCTs) and randomised cross-over trials that included use of activity monitors versus no intervention, another type of intervention, or other activity monitor. Participants were aged 18 years or older with a diagnosis of stroke, in hospital or living in the community. Primary outcome measures were steps per day and time in moderate-to-vigorous intensity activity. Secondary outcomes were sedentary time, time spent in light intensity physical activity, walking duration, fatigue, mood, quality of life, community participation and adverse events. We excluded upper limb monitors that only measured upper limb activity. We followed standard Cochrane methodology to analyse and interpret the data. At least two authors independently screened titles and abstracts for inclusion. We resolved disagreements by consulting a third review author. We extracted the following data from included studies into a standardised template: type of study, participant population, study setting, intervention and co-interventions, time-frame, and outcomes. We graded levels of bias as high, low, or unclear, and assessed the quality of evidence for each outcome using the GRADE approach. We retrieved 28,098 references, from which we identified 29 potential articles. Four RCTs (in 11 reports) met the inclusion criteria.The sample sizes ranged from 27 to 135 (total 245 participants). Time poststroke varied from less than one week (n = 1), to one to three months (n = 2), or a median of 51 months (n = 1). Stroke severity ranged from a median of one to six on the National Institutes of Health Stroke Scale (NIHSS). Three studies were conducted in inpatient rehabilitation, and one was in a university laboratory. All studies compared use of activity monitor plus another intervention (e.g. a walking retraining programme or an inpatient rehabilitation programme) versus the other intervention alone. Three studies reported on the primary outcome of daily step counts.There was no clear effect for the use of activity monitors in conjunction with other interventions on step count in a community setting (mean difference (MD) -1930 steps, 95% confidence interval (CI) -4410 to 550; 1 RCT, 27 participants; very low-quality evidence), or in an inpatient rehabilitation setting (MD 1400 steps, 95% CI -40 to 2840; 2 RCTs, 83 participants; very low-quality evidence). No studies reported the primary outcome moderate-to-vigorous physical activity, but one did report time spent in moderate and vigorous intensity activity separately: this study reported that an activity monitor in addition to usual inpatient rehabilitation increased the time spent on moderate intensity physical activity by 4.4 minutes per day (95% CI 0.28 to 8.52; 1 RCT, 48 participants; low-quality evidence) compared with usual rehabilitation alone, but there was no clear effect for the use of an activity monitor plus usual rehabilitation for increasing time spent in vigorous intensity physical activity compared to usual rehabilitation (MD 2.6 minutes per day, 95% CI -0.8 to 6; 1 RCT, 48 participants; low-quality evidence). The overall risk of bias was low, apart from high-risk for blinding of participants and study personnel. None of the included studies reported any information relating to adverse effects. Only four small RCTs with 274 participants (three in inpatient rehabilitation and one in the community) have examined the efficacy of activity monitors for increasing physical activity after stroke. Although these studies showed activity monitors could be incorporated into practice, there is currently not enough evidence to support the use of activity monitors to increase physical activity after stroke.


Vascular Toxicity in Patients with Cancer: Is There a Recipe to Clarify Treatment? CME.
The acknowledgement of cardiovascular disease as one of the leading causes of mortality and morbidity among cancer survivors is the cornerstone of the growing field of cardio-oncology. Although standardizing treatment for any given disease is often considered ideal, it is important to recognize the value of pursuing a practical and personalized approach when caring for an oncology patient to minimize the risk of treatment-related cardiotoxicity. We hereby discuss a series of cases that illustrate the ways vascular toxicity can manifest in patients with cancer and, when appropriate, provide scientific evidence that supports clinical decision making. We also raise questions about the complex management of these patients while shedding light on future research in this growing field.


Endurance training on low-carbohydrate and grain-based diets: a case study.
To illustrate the effects of low-carbohydrate (LC) and grain-based (GB) diets on body composition, biomarkers, athletic training, and performance in an elite triathlete. The athlete followed 2 dietary interventions for 14 d while maintaining a prescheduled training program. Pre- and postintervention measurements for each diet included plasma and serum samples, resting energy expenditure, body composition, and a performance bike ride. Compared with the GB diet, the LC diet elicited more disruptions to training and unfavorable subjective experiences. Total cholesterol, HDL cholesterol, LDL cholesterol, ratings of perceived exertion, and heart rate were elevated on the LC diet. Blood insulin, resting lactate, postexercise lactate, and C-reactive protein were lowest on the LC diet. The LC diet resulted in both favorable and unfavorable outcomes. The primary observation was a disruption to scheduled training on the LC diet. Researchers should consider how the potential mediating effect of disruptions to training could influence pretest-posttest designs.


p8 and prothymosin alpha: unity is strength.
p8 and prothymosin alpha are two natively unstructured proteins with anti-apoptotic activity. We showed that their interaction results in the formation of a one-to-one heterodimer complex with stable structure. To test whether the heterodimer bears the function previously attributed to both proteins, we monitored the consequences on apoptosis of modulating in vitro the concentrations of both proteins. Overexpression was obtained by transfection of appropriate vectors and inhibition by using specific siRNAs. In all conditions inhibition of apoptosis correlated with the level of the partner with lowest concentration, demonstrating that the anti-apoptotic effect previously attributed to each proteins was in fact borne by the p8/ProTalpha complex, the two proteins, being individually inactive. These results show that the function attributed to a natively unfolded protein might actually belong to a multi-protein complex in which the protein of interest is engaged.


Contraction-related stimuli regulate GLUT4 traffic in C2C12-GLUT4myc skeletal muscle cells.
Muscle contraction stimulates glucose uptake acutely to increase energy supply, but suitable cellular models that faithfully reproduce this complex phenomenon are lacking. To this end, we have developed a cellular model of contracting C(2)C(12) myotubes overexpressing GLUT4 with an exofacial myc-epitope tag (GLUT4myc) and explored stimulation of GLUT4 traffic by physiologically relevant agents. Carbachol (an acetylcholine receptor agonist) induced a gain in cell surface GLUT4myc that was mediated by nicotinic acetylcholine receptors. Carbachol also activated AMPK, and this response was sensitive to the contractile myosin ATPase inhibitor N-benzyl-p-toluenesulfonamide. The gain in surface GLUT4myc elicited by carbachol or by the AMPK activator 5-amino-4-carboxamide-1 beta-ribose was sensitive to chemical inhibition of AMPK activity by compound C and partially reduced by siRNA-mediated knockdown of AMPK catalytic subunits or LKB1. In addition, the carbachol-induced gain in cell surface GLUT4myc was partially sensitive to chelation of intracellular calcium with BAPTA-AM. However, the carbachol-induced gain in cell surface GLUT4myc was not sensitive to the CaMKK inhibitor STO-609 despite expression of both isoforms of this enzyme and a rise in cytosolic calcium by carbachol. Therefore, separate AMPK- and calcium-dependent signals contribute to mobilizing GLUT4 in response to carbachol, providing an in vitro cell model that recapitulates the two major signals whereby acute contraction regulates glucose uptake in skeletal muscle. This system will be ideal to further analyze the underlying molecular events of contraction-regulated GLUT4 traffic.


Allocation of visual attention and anxiety.
Normal individuals high and low in trait-anxiety differ from each other in their attentional functioning. The visual probe experiment was used here to investigate whether subjects with a "repressive coping style" could be distinguished from high-and low-anxious subjects. The three groups were identified by use of the Marlowe-Crowne scale of social desirability and the Spielberger Trait-Anxiety Scale. As predicted, the task clearly distinguished between the groups. High-anxious subjects shifted visual attention toward socially threatening words, represson Wed visual attention away from the same stimuli, and low-anxious subjects showed no consistent pattern of attcntional allocation. These patterns were not observed for stimuli relating to physical threat. It is concluded that individuals reporting low anxiety cannot be considered as a homogeneous group. Theoretically, this is an important finding for the understanding of attentional biases and anxiety.


Niels Stensen: a 17th century scientist with a modern view of brain organization.
In 1665 the Danish scholar Niels Stensen (1638-1686) reached Paris, where he pronounced a discourse on brain anatomy that was to orient neuroscientists for years to come. In his lecture, Stensen rejected ancient speculations about animal spirits and criticized René Descartes and his followers who, despite a poor knowledge of brain anatomy, elaborated complex models to explain the multifaceted function of what he considered the principal organ of the human mind. He advocated the need for studying the brain through a comparative, developmental and pathological convergent approach and called for appropriate dissection methods and accurate illustrations. His own careful anatomical studies permitted him to precisely depict many brain structures. After pioneering works in paleontology and geology, he devoted himself to theology. In 1677 Stensen converted from Lutheranism to Catholicism and, while working relentlessly as a bishop and apostolic vicar in Northern Europe, he died in self-imposed poverty at age 48.


UG4 enhancer-driven GATA-2 and bone morphogenetic protein 4 complementation remedies the CAKUT phenotype in Gata2 hypomorphic mutant mice.
During renal development, the proper emergence of the ureteric bud (UB) from the Wolffian duct is essential for formation of the urinary system. Previously, we showed that expression of transcription factor GATA-2 in the urogenital primordium was demarcated anteroposteriorly into two domains that were regulated by separate enhancers. While GATA-2 expression in the caudal urogenital mesenchyme is controlled by the UG4 enhancer, its more-rostral expression is regulated by UG2. We found that anteriorly displaced budding led to obstructed megaureters in Gata2 hypomorphic mutant mice, possibly due to reduced expression of the downstream effector bone morphogenetic protein 4 (BMP4). Here, we report that UG4-driven, but not UG2-driven, GATA-2 expression in the urogenital mesenchyme significantly reverts the uropathy observed in the Gata2 hypomorphic mutant mice. Furthermore, the data show that transgenic rescue by GATA-2 reverses the rostral outgrowth of the UB. We also provide evidence for a GATA-2-BMP4 epistatic relationship by demonstrating that reporter gene expression from a Bmp4 bacterial artificial chromosome (BAC) transgene is altered in Gata2 hypomorphs; furthermore, UG4-directed BMP4 expression in the mutants leads to reduced incidence of megaureters. These results demonstrate that GATA-2 expression in the caudal urogenital mesenchyme as directed by the UG4 enhancer is crucial for proper development of the urinary tract and that its regulation of BMP4 expression is a critical aspect of this function.


Automatic sorting of multiple unit neuronal signals in the presence of anisotropic and non-Gaussian variability.
Neuronal noise sources and systematic variability in the shape of a spike limit the ability to sort multiple unit waveforms recorded from nervous tissue into their single neuron constituents. Here we present a procedure to efficiently sort spikes in the presence of noise that is anisotropic, i.e., dominated by particular frequencies, and whose amplitude distribution may be non-Gaussian, such as occurs when spike waveforms are a function of interspike interval. Our algorithm uses a hierarchical clustering scheme. First, multiple unit records are sorted into an overly large number of clusters by recursive bisection. Second, these clusters are progressively aggregated into a minimal set of putative single units based on both similarities of spike shape as well as the statistics of spike arrival times, such as imposed by the refractory period. We apply the algorithm to waveforms recorded with chronically implanted micro-wire stereotrodes from neocortex of behaving rat. Natural extension of the algorithm may be used to cluster spike waveforms from records with many input channels, such as those obtained with tetrodes and multiple site optical techniques.