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Bordetella pertussis is the causative agent of whooping cough, a highly contagious respiratory disease. Pertussis toxin (PT), a major virulence factor secreted by B. pertussis, is an AB5-type protein complex topologically related to cholera toxin. The PT protein complex is internalized by host cells and follows a retrograde trafficking route to the endoplasmic reticulum, where it subsequently dissociates. The released enzymatic S1 subunit is then translocated from the endoplasmic reticulum into the cytosol and subsequently ADP-ribosylates the inhibitory alpha-subunits (Gαi) of heterotrimeric G proteins, thus promoting dysregulation of G protein-coupled receptor signaling. However, the mechanistic details of the ADP-ribosylation activity of PT are not well understood. Here, we describe crystal structures of the S1 subunit in complex with nicotinamide adenine dinucleotide (NAD+), with NAD+ hydrolysis products ADP-ribose and nicotinamide, with NAD+ analog PJ34, and with a novel NAD+ analog formed upon S1 subunit crystallization with 3-amino benzamide and NAD+, which we name benzamide amino adenine dinucleotide. These crystal structures provide unprecedented insights into pre- and post-NAD+ hydrolysis steps of the ADP-ribosyltransferase activity of PT. We propose that these data may aid in rational drug design approaches and further development of PT-specific small-molecule inhibitors. [ABSTRACT FROM AUTHOR]

Biomaterial platforms are an integral part of stem cell biomanufacturing protocols. The collective biophysical, biochemical, and cellular cues of the stem cell niche microenvironment play an important role in regulating stem cell fate decisions. Three-dimensional (3D) culture of stem cells within biomaterials provides a route to present biophysical and biochemical stimuli through cell-matrix interactions and cell-cell interactions via secreted biomolecules. Herein, we describe a maleimide-functionalized gelatin (GelMAL) hydrogel that can be crosslinked via thiol-Michael addition click reaction for the encapsulation of sensitive stem cell populations. The maleimide functional units along the gelatin backbone enables gelation via the addition of a dithiol crosslinker without requiring external stimuli (e.g. , UV light, photoinitiator), thereby reducing reactive oxide species generation. Additionally, the versatility of crosslinker selection enables easy insertion of thiol-containing bioactive or bioinert motifs. Hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) were encapsulated in GelMAL, with mechanical properties tuned to mimic the in vivo bone marrow niche. We report the insertion of a cleavable peptide crosslinker that can be degraded by the proteolytic action of Sortase A, a mammalian-inert enzyme. Notably, Sortase A exposure preserves stem cell surface markers, which are an essential metric of hematopoietic activity used in immunophenotyping. This novel GelMAL system enables a route to produce artificial stem cell niches with tunable biophysical properties, intrinsic cell-interaction motifs, and orthogonal addition of bioactive crosslinks. We describe a maleimide-functionalized gelatin hydrogel that can be crosslinked via a thiol-maleimide mediated click reaction to form a stable hydrogel without the production of reactive oxygen species typical in light-based crosslinking. The mechanical properties can be tuned to match the in vivo bone marrow microenvironment for hematopoietic stem cell culture. Additionally, we report inclusion of a peptide crosslinker that can be cleaved via the proteolytic action of Sortase A and show that Sortase A exposure does not degrade sensitive surface marker expression patterns. Together, this approach reduces stem cell exposure to reactive oxygen species during hydrogel gelation and enables post-culture quantitative assessment of stem cell phenotype. [Display omitted] [ABSTRACT FROM AUTHOR]

Background Although previous studies have suggested cocoa products may promote cardiovascular health in the general population, no public data are available from patients receiving care in a national integrated health care system. Objectives We tested the hypothesis that regular chocolate consumption is associated with a lower risk of coronary artery disease (CAD) events among participants of the Million Veteran Program (MVP). Secondary analysis examined if the main hypothesis was observed among participants with type 2 diabetes. Methods We analyzed data from MVP participants who completed the food frequency section of the MVP Lifestyle Survey and were free of CAD at the time of survey completion. CAD events during follow-up (International Statistical Classification of Diseases Ninth Revision codes 410–411 and 413–414, and Tenth Revision codes I20–I25 except I25.2) were assessed using electronic health records. We fitted a Cox proportional hazard model to estimate the RR of CAD. Results Of 188,447 MVP enrollees with survey data, mean ± SD age was 64 ± 12.0 y and 90% were men. For regular chocolate (28.3 g/serving) consumption of <1 serving/mo, 1–3 servings/mo, 1 serving/wk, 2–4 servings/wk, and ≥5 servings/wk, crude incidence rates (per 1000 person-years) for fatal and nonfatal CAD events or coronary procedures were 20.2, 17.5, 16.7, 17.1, and 16.9, respectively, during a mean follow-up of 3.2 y. After adjusting for age, sex, race, and lifestyle factors, the corresponding HRs (95% CIs) were 1.00 (ref), 0.92 (0.87, 0.96), 0.88 (0.83, 0.93), 0.89 (0.84, 0.95), and 0.89 (0.84, 0.96), respectively (P for linear trend < 0.0001). In a secondary analysis of 47,265 diabetics, we did not observe a decreasing trend in CAD mortality among those who consumed ≥1 serving of chocolate a month compared with those who consumed <1 serving/mo. Conclusions Regular chocolate consumption was associated with a lower risk of CAD among veterans, but was not associated with cardiovascular disease risk in veterans with type 2 diabetes. [ABSTRACT FROM AUTHOR]

Limited and inconsistent data are available on the relation between egg consumption and risk of myocardial infarction (MI) and it is unclear if adiposity or type 2 diabetes modifies egg-MI relation. We tested the primary hypothesis that egg consumption is positively associated with incidence of MI among veterans. In secondary analyses, we examined potential effect modification of egg-MI relation by adiposity and type 2 diabetes. We analyzed data collected on 188,267 US veterans who were enrolled in the Million Veteran Program (MVP) from 2011 to 2018. Information on egg consumption was obtained via self-administered food frequency questionnaire and we used electronic health records to identify incident MI. The mean age was 64.4 (SD = 12.0) years and 9.9% of the population were female. We ascertained 10,260 new cases of non-fatal MI during an average follow up of 3.24 years (range: 0.002 to 7.49 y). Hazard ratio (95% CI) for non-fatal MI were 1.00 (ref), 0.93 (0.85–0.1.02), 0.96 (0.87–1.05), 0.98 (0.89–1.07), 1.08 (0.98–1.19), 1.11 (1.00–1.24), and 1.13 (1.00–1.28) for egg consumption of <1/month, 1–3/month, 1/week, 2–4/week, 5–6/week, 1/d, and 2+/d, respectively, controlling for age, sex, race, body mass index, smoking, exercise, alcohol intake, and overall dietary pattern (p non-linear trend 0.019). In secondary analyses, we observed similar results with a composite endpoint including fatal MI, coronary angioplasty and revascularization. Our data showed no association of infrequent consumption of eggs with non-fatal MI but a slightly elevated risk with intake of 1 or more eggs per day among US veterans. [ABSTRACT FROM AUTHOR]

Previous studies of the relationship between fried food consumption and coronary artery disease (CAD) have yielded conflicting results. We tested the hypothesis that frequent fried food consumption is associated with a higher risk of incident CAD events in Million Veteran Program (MVP) participants. Veterans Health Administration electronic health record data were linked to questionnaires completed at MVP enrollment. Self-reported fried food consumption at baseline was categorized: (<1, 1–3, 4–6 times per week or daily). The outcome of interest was non-fatal myocardial infarction or CAD events. We fitted a Cox regression model adjusting for age, sex, race, education, exercise, smoking and alcohol consumption. Of 154,663 MVP enrollees with survey data, mean age was 64 years and 90% were men. During a mean follow-up of approximately 3 years, there were 6,725 CAD events. There was a positive linear relationship between frequency of fried food consumption and risk of CAD (p for trend 0.0015). Multivariable adjusted hazard ratios (95% CI) were 1.0 (ref), 1.07 (1.01–1.13), 1.08 (1.01–1.16), and 1.14 (1.03–1.27) across consecutive increasing categories of fried food intake. In a large national cohort of U.S. Veterans, fried food consumption has a positive, dose-dependent association with CAD. [ABSTRACT FROM AUTHOR]

Observational and clinical trial evidence suggests an inverse association of omega-3 polyunsaturated fatty acids with coronary artery disease (CAD) mortality, although relationships with non-fatal CAD and stroke are less clear. We investigated whether omega-3 fatty acid supplement use and fish intake were associated with incident non-fatal CAD and ischemic stroke among US Veterans. The Million Veteran Program (MVP) is an ongoing nation-wide longitudinal cohort study of US Veterans with self-reported survey, biospecimen, and electronic health record data. Regular use of omega-3 supplements (yes/no) and frequency of fish intake within the past year were assessed using a food frequency questionnaire. Cox proportional hazard models were used to estimate hazard ratios (HR) and 95% confidence intervals (CI) for the associations of omega-3 supplement use and fish intake with incident non-fatal CAD and ischemic stroke, defined from electronic health records using validated algorithms. Multivariable models included demographics, body mass index, education, smoking status, alcohol intake, and exercise frequency. Among 197,761 participants with food frequency data (mean age: 66 ± 12 years, 92% men), 21% regularly took omega-3 supplements and median fish intake was 1 (3–5 ounce) serving/week. Over a median follow-up of 2.9 years for non-fatal CAD and 3.3 years for non-fatal ischemic stroke, we observed 6265 and 4042 incident cases of non-fatal CAD and non-fatal ischemic stroke, respectively. Omega-3 fatty acid supplement use was independently associated with a lower risk of non-fatal ischemic stroke [HR (95% CI): 0.88 (0.81, 0.95)] but not non-fatal CAD [0.99 (0.93, 1.06)]. Fish intake was not independently associated with non-fatal CAD [1.01 (0.94, 1.09) for 1–3 servings/month, 1.03 (0.98, 1.11) for 1 serving/week, 1.02 (0.93, 1.11) for 2–4 servings/week, and 1.15 (0.98, 1.35) for ≥5 servings/week, reference = <1 serving/month, linear p -trend = 0.09] or non-fatal ischemic stroke [0.92 (0.84, 1.00) for 1–3 servings/month, 0.93 (0.85, 1.02) for 1 serving/week, 0.96 (0.86, 1.07) for 2–4 servings/week, and 1.13 (0.93–1.38) for ≥5 servings/week, linear p -trend = 0.16]. Neither omega-3 supplement use, nor fish intake, was associated with non-fatal CAD among US Veterans. While omega-3 supplement use was associated with lower risk of non-fatal ischemic stroke, fish intake was not. Randomized controlled trials are needed to confirm whether omega-3 supplementation is protective against ischemic stroke in a US population. [ABSTRACT FROM AUTHOR]

Abstract Glioblastoma (GBM) is the most common primary malignant brain tumor, with patients exhibiting poor survival (median survival time: 15 months). Difficulties in treating GBM include not only the inability to resect the diffusively-invading tumor cells, but also therapeutic resistance. The perivascular niche (PVN) within the GBM tumor microenvironment contributes significantly to tumor cell invasion, cancer stem cell maintenance, and has been shown to protect tumor cells from radiation and chemotherapy. In this study, we examine how the inclusion of non-tumor cells in culture with tumor cells within a hydrogel impacts the overall gene expression profile of an in vitro artificial perivascular niche (PVN) comprised of endothelial and stromal cells directly cultured with GBM tumor cells within a methacrylamide-functionalized gelatin hydrogel. Using RNA-seq, we demonstrate that genes related to angiogenesis and extracellular matrix remodeling are upregulated in the PVN model compared to hydrogels containing only tumor or perivascular niche cells, while downregulated genes are related to cell cycle and DNA damage repair. Signaling pathways and genes commonly implicated in GBM malignancy, such as MGMT, EGFR , PI3K-Akt signaling, and Ras/MAPK signaling are also upregulated in the PVN model. We describe the kinetics of gene expression within the PVN hydrogels over a course of 14 days, observing the patterns associated with tumor cell-mediated endothelial network co-option and regression. We finally examine the effect of temozolomide, a frontline chemotherapy used clinically against GBM, on the PVN culture. Notably, the PVN model is less responsive to TMZ compared to hydrogels containing only tumor cells. Overall, these results demonstrate that inclusion of cellular and matrix-associated elements of the PVN within an in vitro model of GBM allows for the development of gene expression patterns and therapeutic response relevant to GBM. Graphical abstract Image 1 [ABSTRACT FROM AUTHOR]

FPSO (Floating Production Storage and Off-loading) vessels used for offshore oil and gas production are operated in deep water, often at locations which experience severe wave loading. This paper reports on laboratory experiments on a series of simplified FPSO-shaped bodies, with the aim of understanding more about the wave-structure interaction, particularly the generation of scattered waves. These tests were carried out in the Ocean Basin at Plymouth University’s COAST Laboratory where the effects on the wave-structure interaction of model length, wave steepness and incident wave direction were investigated. All three models had semi-circular ends, separated by a box section for the 2 longer models. Input waves were based on focused wave groups generated using NewWave with an underlying JONSWAP spectrum. A general phase-based harmonic separation method was applied to separate the linear and higher-harmonic wave components of the free-surface elevation surrounding the bodies, and of relevance to the assessment of wave loads. Close to the bow of the model, the highest amplitude scattered waves are observed with the most compact model, and the third- and fourth-harmonics are significantly larger than the equivalent incident bound harmonic components. At the locations close to the stern, the linear harmonic is found to increase as the model length is decreased, although the nonlinear harmonics are similar for all three tested lengths, and the second- and third-harmonics are strongest with the medium length model. The nonlinear scattered waves increase with increasing wave steepness and a second pulse is evident in the higher-order scattered wave fields. As the incident wave angle between the waves and the long axis of the vessel is increased from 0° (head-on) to 20°, the third- and fourth-harmonic scattered waves reduce on the upstream side. These third- and fourth-harmonic diffracted waves should be considered in assessing wave run-up for offshore structure design, and may be relevant to the excitation of ringing-type structural responses in fixed and taut moored structures. [ABSTRACT FROM AUTHOR]

DYT1 dystonia is a movement disorder caused by a trinucleotide deletion (ΔGAG) in DYT1 ( TOR1A ), corresponding to a glutamic acid loss in the C-terminal region of torsinA. Functional alterations in the basal ganglia circuits have been reported in both DYT1 dystonia patients and rodent models. Dyt1 ΔGAG heterozygous knock-in (KI) mice exhibit motor deficits and decreased striatal dopamine receptor 2 (D2R) binding activity, suggesting a malfunction of the indirect pathway. However, the role of the direct pathway in pathogenesis of dystonia is not yet clear. Here, we report that Dyt1 KI mice exhibit significantly decreased striatal dopamine receptor 1 (D1R) binding activity and D1R protein levels, suggesting the alteration of the direct pathway. The decreased D1R may be caused by translational or post-translational processes since Dyt1 KI mice had normal levels of striatal D1R mRNA and a normal number of striatal neurons expressing D1R. Levels of striatal ionotropic glutamate receptor subunits, dopamine transporter, acetylcholine muscarinic M4 receptor and adenosine A2A receptor were not altered suggesting a specificity of affected polytopic membrane-associated proteins. Contribution of the direct pathway to motor-skill learning has been suggested in another pharmacological rat model injected with a D1R antagonist. In the present study, we developed a novel motor skill transfer test for mice and found deficits in Dyt1 KI mice. Further characterization of both the direct and the indirect pathways in Dyt1 KI mice will aid the development of novel therapeutic drugs. [ABSTRACT FROM AUTHOR]

Abstract: Background: The increasing complexity of medical and surgical care often demands comprehensive preoperative work-ups by medical consultants and detailed management recommendations to optimize the patient''s medical ailments before surgery. This article aims to review the available evidence and discuss elements of the preoperative medical consultation that are of particular relevance to surgeons. Methods: The authors performed a comprehensive review of the available medical literature and guidelines pertaining to preoperative medical management and present a narrative summary of their findings. Results: Although many preoperative recommendations are based on expert opinion and consensus, a growing number of studies and clinical practice guidelines provides direction as to what is the best management of patients with medical problems before surgery. Conclusions: Collaborative efforts and adequate communication between the medical and surgical teams are necessary to ensure appropriate patient management before surgery. Although higher-risk patients may require thorough preoperative evaluations, extensive routine investigations may not always be necessary. [Copyright &y& Elsevier]

Abstract: Myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonia. DYT11 M-D is caused by mutations in SGCE which codes for ɛ-sarcoglycan. SGCE is maternally imprinted and paternally expressed. Abnormal nuclear envelope has been reported in mouse models of DYT1 generalized torsion dystonia. However, it is not known whether similar alterations occur in DYT11 M-D. We developed a mouse model of DYT11 M-D using paternally inherited Sgce heterozygous knockout (Sgce KO) mice and reported that they had myoclonus and motor coordination and learning deficits in the beam-walking test. However, the specific brain regions that contribute to these phenotypes have not been identified. Since ɛ-sarcoglycan is highly expressed in the cerebellar Purkinje cells, here we examined the nuclear envelope in these cells using a transmission electron microscope and found that they are abnormal in Sgce KO mice. Our results put DYT11 M-D in a growing family of nuclear envelopathies. To analyze the effect of loss of ɛ-sarcoglycan function in the cerebellar Purkinje cells, we produced paternally inherited cerebellar Purkinje cell-specific Sgce conditional knockout (Sgce pKO) mice. Sgce pKO mice showed motor learning deficits, while they did not show abnormal nuclear envelope in the cerebellar Purkinje cells, robust motor deficits, or myoclonus. The results suggest that ɛ-sarcoglycan in the cerebellar Purkinje cells contributes to the motor learning, while loss of ɛ-sarcoglycan in other brain regions may contribute to nuclear envelope abnormality, myoclonus and motor coordination deficits. [Copyright &y& Elsevier]

Abstract: DYT1 early-onset generalized torsion dystonia is an inherited movement disorder associated with mutations in DYT1 that codes for torsinA protein. The most common mutation seen in this gene is a trinucleotide deletion of GAG. We previously reported a motor control deficit on a beam-walking task in our Dyt1 ΔGAG knock-in heterozygous mice. In this report we show the reversal of this motor deficit with the anticholinergic trihexyphenidyl (THP), a drug commonly used to treat movement problems in dystonia patients. THP also restored the reduced corticostriatal long-term depression (LTD) observed in these mice. Corticostriatal LTD has long been known to be dependent on D2 receptor activation. In this mouse model, striatal D2 receptors were expressed at lower quantities in comparison to wild-type mice. Furthermore, the mice were also partially resistant to FPL64176, an agonist of L-type calcium channels that have been previously reported to cause severe dystonic-like symptoms in wild-type mice. Our findings collectively suggest that altered communication between cholinergic interneurons and medium spiny neurons is responsible for the LTD deficit and that this synaptic plasticity modification may be involved in the striatal motor control abnormalities in our mouse model of DYT1 dystonia. [Copyright &y& Elsevier]

Introduction: Gastrointestinal leak is a dreaded complication after esophagectomy. Conventional treatments for leak include conservative therapy, surgical reoperation, and even complete gastrointestinal (GI) diversion. The aim of this study was to evaluate the impact of endoluminal stenting in the management of esophagogastric leak after esophagectomy.Methods: Data on 18 (11.3%) of 160 patients who developed postoperative leaks after minimally invasive esophagectomy were reviewed. Indications for esophagectomy included carcinoma (n = 14), Barrett's with high-grade dysplasia (n = 3), and benign stricture (n = 1). Neoadjuvant therapy was used in 57.1% of patients with carcinoma. The first nine patients underwent conventional treatments for leak whereas the latter nine patients underwent endoscopic esophageal covered stenting as primary therapy. There were 5 cervical and 13 intrathoracic anastomotic leaks. Main outcome measures included patient characteristics, types of treatment, length of hospital stay, morbidity, and mortality.Results: Subjects were 16 males and 2 females with a mean age of 66 years. In the conventional treatment group, leaks were treated with neck drainage (n = 4), GI diversion (n = 2), and thoracoscopic drainage with or without repair or T-tube placement (n = 3). In the endoscopy group, all leaks were treated with endoscopic covered stenting with or without percutaneous drainage (n = 9). Control of leaks occurred in 89% of patients in the conventional treatment group vs. 100% of patients in the endoscopic stenting group. Three patients in the conventional treatment group (33%) required esophageal diversion compared to none of the patients in the endoscopy group. The 60-day or in-hospital mortality was 0% for both groups.Conclusion: In our clinical practice, there has been a shift in the management of esophagogastric anastomotic leaks to nonsurgical therapy using endoscopic esophageal covered stenting. Endoluminal stenting is a safe and effective alternative in the management of GI leaks. [ABSTRACT FROM AUTHOR]

The Fc-α receptor (FcαR/CD89) is involved in IgA complex formation and may affect the development of IgA nephropathy (IgAN). In this study, we tested the genetic variations of the CD89 gene in relation to disease susceptibility in IgAN and the expression of soluble CD89 (sCD89) in sera of patients with IgAN and in controls. There was a significant difference between the levels of sCD89-IgA complexes, measured by sandwich enzyme-linked immunosorbent assay (ELISA), in 177 patients with IgAN with and without disease progression at the time of first diagnosis. No such difference was found in 42 patients with other renal diseases. The patients with IgAN without disease progression had stable but high levels of sCD89 over 5-15 years of follow-up in contrast to stable but low levels of sCD89 in the disease progression group. Moreover, levels of sCD89 complexes were correlated with one of the five CD89 genetic variants in 212 patients with IgAN and 477 healthy Caucasians; the single-nucleotide polymorphism (SNP) rs11084377 was significantly associated with a lower expression of sCD89. However, no association between CD89 gene polymorphisms and susceptibility to IgAN was detected. Thus, we found an association between the levels of sCD89-IgA complexes in serum and the severity of IgAN, and a possible genetic component in regulating the production or expression of sCD89. [ABSTRACT FROM AUTHOR]

Background Serum creatinine is a delayed marker of acute kidney injury (AKI). Our purpose is to discover and validate novel early urinary biomarkers of AKI after cardiac surgery. Study Design Diagnostic test study. Setting & Participants Children undergoing cardiopulmonary bypass surgery. The test set included 15 participants with AKI and 15 matched controls (median age, 1.5 year) of 45 participants without AKI. The validation set included 365 children (median age, 1.9 year). Index Tests Biomarkers identified using proteomic profiling: α1-microglobulin, α1-acid glycoprotein, and albumin. Reference Test AKI, defined as ≥50% increase in serum creatinine level from baseline within 3 days of surgery. Results Proteomic profiling using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) showed 3 protein peaks that appeared consistently within 2 hours in children who developed AKI after cardiopulmonary bypass surgery. The proteins were identified as a1-microglobulin, a1-acid glycoprotein, and albumin. Using clinical assays, results were confirmed in a test set and validated in an independent prospective cohort. In the validation set, 135 (37%) developed AKI, in whom there was a progressive increase in urinary biomarker concentrations with severity of AKI. Areas under the curve for urinary a1-microglobulin, a1-acid glycoprotein, and albumin at 6 hours after cardiac surgery were 0.84 (95% CI, 0.79-0.89), 0.87 (95% CI, 0.83-0.91), and 0.76 (95% CI, 0.71-0.81), respectively. Participants with increasing quartiles of biomarkers showed increasing lengths of hospital stays and durations of AKI (P < 0.001). Limitations Single-center study of children with normal kidney function at recruitment. The SELDI-TOF MS technique has limited sensitivity for the detection of proteins greater than the 20-kDa range. Conclusions Urinary α1-microglobulin, α1-acid glycoprotein, and albumin represent early, accurate, inexpensive, and widely available biomarkers of AKI after cardiac surgery. They also offer prognostic information about the duration of AKI and length of hospitalization after cardiac surgery. [ABSTRACT FROM AUTHOR]

Abstract: Background: Obesity is a well-known risk factor for the development of coronary heart disease (CHD). The aim of the present study was to examine the differences in the 10-year CHD risk with increasing severity of obesity in men and women participating in the latest National Health and Nutrition Examination Survey. Methods: Data from a representative sample of 12,500 U.S. participants in the National Health and Nutrition Examination Survey from 1999 to 2006 were reviewed. The Framingham risk score was calculated for men and women according to a body mass index (BMI) of <25.0, 25.0–29.9, 30.0–34.9, and ≥35.0 kg/m2. Results: The prevalence of those with hypertension increased with an increasing BMI, from 24% for a BMI <25.0 kg/m2 to 54% for a BMI of ≥35.0 kg/m2. The prevalence of an abnormal total cholesterol level (>200 mg/dL) increased from 40% for a BMI <25.0 kg/m2 to 48% for a BMI of ≥35.0 kg/m2. The 10-year CHD risk for men increased from 3.1% for a BMI <25.0 kg/m2 to a peak of 5.6% for a BMI of 30.0–34.9 kg/m2. The 10-year CHD risk for women increased from .8% for a BMI <25.0 kg/m2 to a peak of 1.5% for a BMI of ≥35.0 kg/m2. Both diabetes and hypertension were independent risk factors for an increasing CHD risk. Conclusions: The 10-year CHD risk, calculated using the Framingham risk score, substantially increased with an increasing BMI. An important implication from our findings is the need to implement surgical and medical approaches to weight reduction to reduce the effect of morbidity and mortality from CHD on the U.S. healthcare system. [ABSTRACT FROM AUTHOR]

Abstract: DYT1 dystonia is caused by a trinucleotide deletion of GAG (ΔGAG) in DYT1, which codes for torsinA. A previous epidemiologic study suggested an association of DYT1 ΔGAG mutation with early-onset recurrent major depression. However, another study reported no significant association with depression, but instead showed an association with anxiety and dystonia. In this study, we analyzed these related behaviors in Dyt1 ΔGAG heterozygous knock-in mice. The knock-in mice showed a subtle anxiety-like behavior but did not show depression-like behaviors. The mutant mice also displayed normal sensorimotor gating function in a prepulse inhibition test. While normal hippocampus-dependent contextual fear memory and hippocampal CA1 long-term potentiation (LTP) were observed, the knock-in mice exhibited an enhancement in the formation of cued fear memories. Anatomical analysis indicated that the number of c-fos positive cells was significantly increased while the size of the central nucleus of the amygdala (CE) was significantly reduced in the knock-in mice. These results suggest that the Dyt1 ΔGAG mutation increased the activity of the CE and enhanced the acquisition of the cued fear memory. [Copyright &y& Elsevier]

Background: Obesity has been linked with a chronic state of inflammation which may be involved in the development of metabolic syndrome, cardiovascular disease, non-alcoholic steatohepatitis, and even cancer. The objective of this study was to examine the association between obesity class and levels of inflammatory biomarkers from men and women who participated in the 1999-2004 National Health and Nutrition Examination Survey (NHANES).Methods: Serum concentrations of C-reactive protein (CRP) and fibrinogen were measured among US participants of the 1999-2004 NHANES. We examined biomarker levels across different weight classes with normal weight, overweight, and obesity classes 1, 2, and 3 were defined as BMI of < 25.0, 25.0-29.9, 30.0-34.9, 35.0-39.9, and > or = 40.0, respectively.Results: With CRP levels for normal weight individuals as a reference, CRP levels nearly doubled with each increase in weight class: +0.11 mg/dl (95% CI, 0.06-0.16) for overweight, +0.21 mg/dl (95% CI, 0.16-0.27) for obesity class 1, +0.43 mg/dl (95% CI, 0.26-0.61) for obesity class 2, and +0.73 mg/dl (95% CI, 0.55-0.90) for obesity class 3. With normal weight individuals as a reference, fibrinogen levels increase with increasing weight class and were highest for obesity class 3 individuals, +93.5 mg/dl (95% CI, 72.9-114.1). Individuals with hypertension or diabetes have higher levels of CRP and fibrinogen levels compared to individuals without hypertension or diabetes, even when stratified according to BMI.Conclusions: There is a direct association between increasing obesity class and the presence of obesity-related comorbidities such as diabetes and hypertension with high levels of inflammatory biomarkers. [ABSTRACT FROM AUTHOR]

Abstract: Engineering tissue similar in structure to their natural equivalents is a major challenge and crucial to function. Despite attempts to engineer skeletal muscle, it is still difficult to effectively mimic tissue architecture. Rigid scaffolds can guide cell alignment but have the critical drawback of hindering mechanical function of the resultant tissue. We present a method for creating highly ordered tissue-only constructs by using rigid microtopographically patterned surfaces to first guide myoblast alignment, followed by transfer of aligned myotubes into a degradable hydrogel and self-organization of the ordered cells into a functional, 3-dimensional, free-standing construct independent of the initial template substrate. Histology revealed an intracellular organization resembling that of native muscle. Aligned cell constructs exhibited a 2-fold increase in peak force production compared to controls. Effective specific force, or force normalized over cross-sectional area, was increased by 23%. This template, transfer, and self-organization strategy is envisioned to be broadly useful in improving construct function and clinical applicability for highly ordered tissues like muscle. [Copyright &y& Elsevier]

Abstract: Traditional cell culture substrates consist of static, flat surfaces although in vivo, cells exist on various dynamic topographies. We report development of a reconfigurable microtopographical system compatible with cell culture that is comprised of reversible wavy microfeatures on poly(dimethylsiloxane). Robust reversibility of the wavy micropattern is induced on the cell culture customized substrate by first plasma oxidizing the substrate to create a thin, brittle film on the surface and then applying and releasing compressive strain, to introduce and remove the microfeatures, respectively. The reversible topography was able to align, unalign, and realign C2C12 myogenic cell line cells repeatedly on the same substrate within 24 h intervals, and did not inhibit cell differentiation. The flexibility and simplicity of the materials and methods presented here provide a broadly applicable capability by which to investigate and compare dynamic cellular processes not yet easily studied using conventional in vitro culture substrates. [Copyright &y& Elsevier]

Recent foodborne outbreaks implicating spinach and lettuce have increased consumer concerns regarding the safety of fresh produce. While the most common commercial antimicrobial intervention for fresh produce is wash water containing 50 to 200 ppm chlorine, this study compares the effectiveness of acidified sodium chlorite, chlorine, and acidic electrolyzed water for inactivating Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes inoculated onto leafy greens. Fresh mixed greens were left uninoculated or inoculated with approximately 6 log CFU/g of E. coli O157:H7, Salmonella, and L. monocytogenes and treated by immersion for 60 or 90 s in different wash solutions (1:150, wt/vol), including 50 ppm of chlorine solution acidified to pH 6.5, acidic electrolyzed water (pH 2.1 ± 0.2, oxygen reduction potential of 1,100 mV, 30 to 35 ppm of free chlorine), and acidified sodium chlorite (1,200 ppm, pH 2.5). Samples were neutralized and homogenized. Bacterial survival was determined by standard spread plating on selective media. Each test case (organism x treatment x time) was replicated twice with five samples per replicate. There was no difference (P ≥ 0.05) in the time of immersion on the antimicrobial effectiveness of the treatments. Furthermore, there was no difference (P ≥ 0.05) in survival of the three organisms regardless of treatment or time. Acidified sodium chlorite, resulted in reductions in populations of 3 to 3.8 log CFU/g and was more effective than chlorinated water (2.1 to 2.8 log CFU/g reduction). These results provide the produce industry with important information to assist in selection of effective antimicrobial strategies. [ABSTRACT FROM AUTHOR]

Abstract: The DYT1 gene containing a trinucleotide deletion (ΔGAG) is linked to early-onset dystonia, a neurological movement disorder of involuntary muscle contractions. To understand DYT1''s contribution to dystonia, we produced and analyzed Dyt1 knockdown (KD) mice that expressed a reduced level of torsinA protein encoded by Dyt1. Knockdown mice exhibited deficits in motor control and a decreased trend in dopamine with a significant reduction in 3,4-dihydroxyphenylacetic acid. These alterations are similar to those displayed by previously reported Dyt1 ΔGAG knockin heterozygous mice, suggesting that the partial loss of torsinA function contributes to the pathology of the disease. [Copyright &y& Elsevier]

Abstract: Tissue-engineered muscle is a viable option for tissue repair, though presently technologies are not developed enough to produce tissue in vitro identical to that in vivo. One important step in generating accurate engineered muscle is to mimic natural muscle architecture. Skeletal muscle is composed of fibrils whose organization defines functionality. In musculoskeletal myogenesis, aligning myoblasts in preparation for myotube formation is a crucial step. The ability to efficiently organize myoblasts to form aligned myotubes in vitro would greatly benefit efforts in muscle tissue engineering. This paper reports alignment of prefused and differentiated skeletal muscle cells in vitro by use of continuous micropatterned wavy silicone surfaces, with features sized 3, 6 and 12μm in periodicity. Wave features with 6μm periodicity produced the most healthy, aligned myoblasts. Alignment was found to be a function of plating density. Further growth on these substrates with aligned myoblasts promoted fusion, yielding healthy aligned myotubes. This method will be useful for applications in which differentiated myogenic cells need to be aligned unidirectionally as in the development of engineered muscle. [Copyright &y& Elsevier]

Rationale and Objectives: MRI has shown promise in assessing breast cancer patients undergoing neoadjuvant chemotherapy. Computer-aided detection (CAD) for MRI can automatically display tumor enhancement parameters. This study was performed to determine the utility of CAD applied to breast MRI in this patient population. Materials and Methods: Fifteen patients with 16 newly diagnosed locally advanced breast cancers were evaluated with MRI before and after neoadjuvant chemotherapy. CAD assessments, including presence or absence of significant enhancement, enhancement profiles, and maximum sizes, were recorded. Pre-chemotherapy and post-chemotherapy enhancement profiles were compared. Sizes were compared to those measured by the radiologist and at final pathology. Results: Prior to chemotherapy, all tumors demonstrated CAD-assessed significant enhancement. Following chemotherapy, 7/16 tumors showed no residual significant enhancement, but all had residual disease at pathology. In those patients with residual enhancement, comparison of the post-chemotherapy to pre-chemotherapy CAD enhancement profiles showed a significant decrease in percentage of washout enhancement (P = 0.0147) in patients with less than 5 mm of residual microscopic disease. Radiologist-measured tumor sizes demonstrated better correlation with sizes at pathology (r = 0.60) than did CAD-generated tumor sizes (r = 0.32). Conclusion: CAD may be helpful in assessing changes in MRI enhancement profiles of tumors following chemotherapy. However, CAD-assessed significant enhancement following chemotherapy can be falsely negative for residual malignancy, and CAD tumor sizes are less accurate than those measured by the radiologist in predicting size of residual malignancy. CAD may complement but should not replace the radiologist’s assessment of tumors in this patient population. [Copyright &y& Elsevier]

Interleukin-8 (IL-8) may be important in psoriasis as it is expressed in the stratum granulosum, attracts polymorphonuclear cells, and stimulates angiogenesis and keratinocyte mitogenesis. To study intrinsic cutaneous factors in psoriasis, we constructed skin equivalents from psoriatic or adult control fibroblasts with normal foreskin keratinocytes. IL-8 levels were measured in supernatants by enzyme-linked immunosorbent assay and in skin equivalents by immuno- histochemistry and in situ hybridization, JL-8 was highly induced in skin equivalents compared to cells grown alone. Epidermal stratification varied among fibroblast lines and was correlated with IL-8 levels, but lesional and nonlesional psoriatic skin equivalents from the same donor were similar. Six fibroblast lines (two psoriasis lesion and four normal) supported only monolayers, while 12 lines (seven psoriasis lesion and five normal) produced stratification. Mean IL-8 levels were significantly lower in dermal equivalents of the first group than the second (0.78 ± 0.40 vs 3.93 ± 2.83 ng per ml, mean ± SD, p = 0.01, analysis of variance). Significantly more IL-S was secreted by psoriatic than normal fibroblast skin equivalents over 14 d (p = 0.015) with greatest differences at 1 and 4 d. Psoriatic IL-8 levels peaked first and remained increased, IL-8 protein and mRNA were initially strongest in dermal fibroblasts, and at the dermal-epidermal interface. Diffuse epidermal expression was replaced by accentuation in the stratum granulosum. Psoriatic skin equivalents were thicker, had more intense IL-8 staining, and developed invagination. We hypothesize that an IL-8 paracrine loop between fibroblasts and keratinocytes may play a key role in epidermal regeneration in the skin equivalent, in normal wound healing, and in the determination of an intrinsic psoriatic wound-healing phenotype. [ABSTRACT FROM AUTHOR]

Abstract: Our research suggests that in 2050, 80% of total U.S. electricity demand could be supplied by renewable electricity technologies that are commercially available today at a cost similar to or lower than published estimates for other clean energy scenarios. In order to achieve this goal, the U.S. power system will need to evolve toward increased efficiency and system flexibility – in part enabled by grid expansion and new operating procedures. Adjustments in business models, market rules, and regulatory regimes may also be needed to handle these high levels of renewables with their different financial and operating characteristics. This short Report Review highlights aspects of policy, regulation, finance, markets and operations that can help enable high penetration renewable energy electricity generation futures. It uses analytical results from the NREL Renewable Electricity Futures (REF) Study [1] as a basis for discussion. As technical issues have been shown not to be key impediments for this pathway at the hourly level for the bulk system, we focus on other aspects of public and private decision-making. We conclude by describing how the REF might inform future research and development by the scientific community. [Copyright &y& Elsevier]

Tumor-associated macrophages (TAMs) play an important role in tumor immunity and comprise of subsets that have distinct phenotype, function, and ontology. Transcriptomic analyses of human medulloblastoma, the most common malignant pediatric brain cancer, showed that medulloblastomas (MBs) with activated sonic hedgehog signaling (SHH-MB) have significantly more TAMs than other MB subtypes. Therefore, we examined MB-associated TAMs by single-cell RNA sequencing of autochthonous murine SHH-MB at steady state and under two distinct treatment modalities: molecular-targeted inhibitor and radiation. Our analyses reveal significant TAM heterogeneity, identify markers of ontologically distinct TAM subsets, and show the impact of brain microenvironment on the differentiation of tumor-infiltrating monocytes. TAM composition undergoes dramatic changes with treatment and differs significantly between molecular-targeted and radiation therapy. We identify an immunosuppressive monocyte-derived TAM subset that emerges with radiation therapy and demonstrate its role in regulating T cell and neutrophil infiltration in MB. [Display omitted] • Sonic Hedgehog (SHH) subgroup of medulloblastoma (MB) recruits diverse macrophages • Radiation or molecular-targeted therapy alters macrophage distribution in SHH-MB • Radiation recruits immunosuppressive monocyte-derived macrophages (TAMoMacs) in SHH-MB • Radiation-induced TAMoMacs regulate CD8 T cell and neutrophil numbers in SHH-MB Dang et al. show that the sonic hedgehog subgroup of medulloblastoma (SHH-MB) contains macrophages derived from microglia and circulating monocytes. Radiation therapy, but not treatment targeting the SHH pathway, led to recruitment of immunosuppressive monocyte-derived macrophages that reduced T cells and neutrophils in the tumor microenvironment. [ABSTRACT FROM AUTHOR]

The vasculature is a key component of the tissue microenvironment. Traditionally known for its role in providing nutrients and oxygen to surrounding cells, the vasculature is now also acknowledged to provide signaling cues that influence biological outcomes in regeneration and disease. These cues come from the cells that comprise vasculature, as well as the dynamic biophysical and biochemical properties of the surrounding extracellular matrix that accompany vascular development and remodeling. In this review, we illustrate the larger role of the vasculature in the context of regenerative biology and cancer progression. We describe cellular, biophysical, biochemical, and metabolic components of vascularized microenvironments. Moreover, we provide an overview of multidimensional angiogenic biomaterials that have been developed to promote therapeutic vascularization and regeneration, as well as to mimic elements of vascularized microenvironments as a means to uncover mechanisms by which vasculature influences cancer progression and therapy. [ABSTRACT FROM AUTHOR]

Background: Environmental cues associated with cocaine evoke craving and seeking. This process, termed cue reactivity, is a critical element of cocaine addiction. Although glutamatergic neurotransmission has been implicated in this effect of cocaine, the precise subtype and localization in the brain of the glutamatergic receptor critical for cocaine cue reactivity is not well-understood. Methods: We used a conditional N-methyl-D-aspartic acid receptor (NMDAR) knockout mouse whose NMDAR gene was deleted by Cre expression restricted to striatal neurons. To evaluate the role of NMDAR in cocaine cue reactivity, conditional knockout mice and control mice (n = 5–8/group) were conditioned for place preference with cocaine (5 and 10 mg/kg SC) for 3 days. Their subsequent place preference was examined in a drug-free state. Results: Although control mice developed cocaine conditioned place preference, mice deficient for NMDAR on striatal neurons failed to develop conditioned place preference. Conclusions: The NMDAR on striatal neurons is essential for the development of cocaine cue reactivity in the place conditioning paradigm. Our finding identifies a brain region whose constitutive NMDAR level serves as a determinant for susceptibility to this aspect of cocaine addiction. [Copyright &y& Elsevier]