Phase-dependence of response curves to deep brain stimulation and their relationship: from essential tremor patient data to a Wilson-Cowan model.
Essential tremor manifests predominantly as a tremor of the upper limbs. One therapy option is high-frequency deep brain stimulation, which continuously delivers electrical stimulation to the ventral intermediate nucleus of the thalamus at about 130 Hz. Constant stimulation can lead to side effects, it is therefore desirable to find ways to stimulate less while maintaining clinical efficacy. One strategy, phase-locked deep brain stimulation, consists of stimulating according to the phase of the tremor. To advance methods to optimise deep brain stimulation while providing insights into tremor circuits, we ask the question: can the effects of phase-locked stimulation be accounted for by a canonical Wilson-Cowan model? We first analyse patient data, and identify in half of the datasets significant dependence of the effects of stimulation on the phase at which stimulation is provided. The full nonlinear Wilson-Cowan model is fitted to datasets identified as statistically significant, and we show that in each case the model can fit to the dynamics of patient tremor as well as to the phase response curve. The vast majority of top fits are stable foci. The model provides satisfactory prediction of how patient tremor will react to phase-locked stimulation by predicting patient amplitude response curves although they were not explicitly fitted. We also approximate response curves of the significant datasets by providing analytical results for the linearisation of a stable focus model, a simplification of the Wilson-Cowan model in the stable focus regime. We report that the nonlinear Wilson-Cowan model is able to describe response to stimulation more precisely than the linearisation.


Annular remodeling in chronic ischemic mitral regurgitation: ring selection implications.
More precise understanding of annular remodeling in the evolution of chronic ischemic mitral regurgitation is needed to provide a more rational basis for optimal annuloplasty ring sizing and selection as well as the design of new reparative techniques. Three-dimensional in vivo data describing these geometric perturbations however are lacking. Using an ovine model of chronic myocardial infarction we determined the three-dimensional distortions of the mitral annulus associated with the development of chronic ischemic mitral regurgitation. Ten sheep underwent placement of radiopaque markers on the left ventricle and mitral annulus as well as placement of snares around the second and third obtuse marginal coronary arteries. After 8 days biplane cinefluoroscopy provided three-dimensional marker data and snare occlusion created an inferior infarction. After 7 more weeks the animals were studied again. Severity of mitral regurgitation increased (0.6 +/- 0.5 to 2.5 +/- 0.7). Septal-lateral (2.99 +/- 0.20 cm to 3.64 +/- 0.35 cm, maximum dimension) and commissure-commissure (3.71 +/- 0.32 cm to 4.40 +/- 0.30 cm) mitral annular diameters and the lengths of the muscular (7.77 +/- 0.39 cm to 9.51 +/- 0.72 cm) and fibrous annular perimeters (3.36 +/- 0.37 cm to 3.85 +/- 0.39 cm, p < 0.0001 for all) increased while the height of the annular "saddle horn" above a best-fit plane fell (0.73 +/- 0.52 cm to 0.57 +/- 0.42 cm, minimum dimension, p = 0.01). These three-dimensional in vivo data reflect annular remodeling in chronic ischemic mitral regurgitation and suggest that mitral repair in this context should be aimed at preventing further lengthening of the intertrigonal distance, reducing the septal-lateral annular diameter to reestablish adequate leaflet coaptation, and restoring the saddle shape of the annulus.


Factors affecting the regulation of pacing: current perspectives.
During prolonged dynamic and rhythmic exercise, muscular pain and discomfort arises as a result of an increased concentration of deleterious metabolites. Sensed by peripheral nociceptors and transmitted via afferent feedback to the brain, this provides important information regarding the physiological state of the muscle. These sensations ultimately contribute to what is termed "exercise-induced pain". Despite being well recognized by athletes and coaches, and suggested to be integral to exercise performance, this construct has largely escaped attention in experimental work. This perspective article highlights the current understanding of pacing in endurance performance, and the causes of exercise-induced pain. A new perspective is described, which proposes how exercise-induced pain may be a contributing factor in helping individuals to regulate their work rate during exercise and thus provides an important construct in pacing.


Viral Metagenomic Analysis of Cerebrospinal Fluid from Patients with Acute Central Nervous System Infections of Unknown Origin, Vietnam.
Central nervous system (CNS) infection is a serious neurologic condition, although the etiology remains unknown in >50% of patients. We used metagenomic next-generation sequencing to detect viruses in 204 cerebrospinal fluid (CSF) samples from patients with acute CNS infection who were enrolled from Vietnam hospitals during 2012-2016. We detected 8 viral species in 107/204 (52.4%) of CSF samples. After virus-specific PCR confirmation, the detection rate was lowered to 30/204 (14.7%). Enteroviruses were the most common viruses detected (n = 23), followed by hepatitis B virus (3), HIV (2), molluscum contagiosum virus (1), and gemycircularvirus (1). Analysis of enterovirus sequences revealed the predominance of echovirus 30 (9). Phylogenetically, the echovirus 30 strains belonged to genogroup V and VIIb. Our results expanded knowledge about the clinical burden of enterovirus in Vietnam and underscore the challenges of identifying a plausible viral pathogen in CSF of patients with CNS infections.


A tadpole-shaped gene carrier with distinct phase segregation in a ternary polymeric micelle.
A distinct tadpole-shaped nanostructure characterized by a spherical head and an extended shaft was identified in a single plasmid DNA (pDNA)-based polymeric micelle. The tadpole-shaped structure was constructed by adding anionic chondroitin sulfate (CS) to the rod-shaped polyplex micelle containing a single pDNA molecule packaged by the PEG-polycation block copolymer through their electrostatic self-assembly. The complex consequently developed a novel structure composed of segregated domains of the CS-rich inflated head and CS-poor folded DNA tail. Hence, this tadpole structure can be regarded as evidence that distinct phase segregation occurred in a single polymeric micelle containing pDNA.


Renal Manifestations of Tuberous Sclerosis Complex: Key Findings From the Final Analysis of the TOSCA Study Focussing Mainly on Renal Angiomyolipomas.
Renal angiomyolipomas are one of the most common renal manifestations in patients with tuberous sclerosis complex (TSC), with potentially life-threatening complications and a poor prognosis. Despite the considerable progress in understanding TSC-associated renal angiomyolipomas, there are no large scale real-world data. The aim of our present study was to describe in detail the prevalence and outcome of renal angiomyolipomas in patients with TSC, enrolled into the TuberOus SClerosis registry to increase disease Awareness (TOSCA) from 170 sites across 31 countries worldwide. We also sought to evaluate the relationship of TSC-associated renal angiomyolipomas with age, gender and genotype. The potential risk factors for renal angiomyolipoma-related bleeding and chronic kidney disease (CKD) were studied in patients who participated in the TOSCA renal angiomyolipoma substudy. Of the 2,211 eligible patients, 1,062 (48%) reported a history of renal angiomyolipomas. The median age of TSC diagnosis for the all subjects (n = 2,211) was 1 year. The median age of diagnosis of renal angiomyolipoma in the 1,062 patients was 13 years. Renal angiomyolipomas were significantly more prevalent in female patients (p < 0.0001). Rates of angiomyolipomas >3 cm (p = 0.0119), growing lesions (p = 0.0439), and interventions for angiomyolipomas (p = 0.0058) were also higher in females than males. Pre-emptive intervention for renal angiomyolipomas with embolisation, surgery, or mammalian target of rapamycin (mTOR) inhibitor may have abolished the gender difference in impaired renal function, hypertension, and other complications. The rate of interventions for angiomyolipomas was less common in children than in adults, but interventions were reported in all age groups. In the substudy of 76 patients the complication rate was too low to be useful in predicting risk for more severe CKD. In addition, in this substudy no patient had a renal hemorrhage after commencing on an mTOR inhibitor. Our findings confirmed that renal angiomyolipomas in subjects with TSC1 mutations develop on average at the later age, are relatively smaller in size and less likely to be growing; however, by age 40 years, no difference was observed in the percentage of patients with TSC1 and TSC2 mutations needing intervention. The peak of appearance of new renal angiomyolipomas was observed in patients aged between 18 and 40 years, but, given that angiomyolipomas can occur later, lifelong surveillance is necessary. We found that pre-emptive intervention was dramatically successful in altering the outcome compared to historical controls; with high pre-emptive intervention rates but low rates of bleeding and other complications. This validates the policy of surveillance and pre-emptive intervention recommended by clinical guidelines.


Recounting the FANTOM CAGE-Associated Transcriptome.
Long noncoding RNAs (lncRNAs) have emerged as key coordinators of biological and cellular processes. Characterizing lncRNA expression across cells and tissues is key to understanding their role in determining phenotypes, including human diseases. We present here FC-R2, a comprehensive expression atlas across a broadly defined human transcriptome, inclusive of over 109,000 coding and noncoding genes, as described in the FANTOM CAGE-Associated Transcriptome (FANTOM-CAT) study. This atlas greatly extends the gene annotation used in the original recount2 resource. We demonstrate the utility of the FC-R2 atlas by reproducing key findings from published large studies and by generating new results across normal and diseased human samples. In particular, we (a) identify tissue-specific transcription profiles for distinct classes of coding and noncoding genes, (b) perform differential expression analysis across thirteen cancer types, identifying novel noncoding genes potentially involved in tumor pathogenesis and progression, and (c) confirm the prognostic value for several enhancer lncRNAs expression in cancer. Our resource is instrumental for the systematic molecular characterization of lncRNA by the FANTOM6 Consortium. In conclusion, comprised of over 70,000 samples, the FC-R2 atlas will empower other researchers to investigate functions and biological roles of both known coding genes and novel lncRNAs.


Identifying tissue-specific signal variation in MALDI mass spectrometric imaging by use of an internal standard.
Generating analyte-specific distribution maps of compounds in a tissue sample by matrix-assisted laser desorption/ionization (MALDI) mass spectrometric imaging (MSI) has become a useful tool in numerous areas across the biological sciences. Direct analysis of the tissue sample provides MS images of an analyte's distribution with minimal sample pretreatment. The technique, however, suffers from the inability to account for tissue-specific variations in ion signal. The variation in the makeup of different tissue types can result in significant differences in analyte extraction, cocrystallization, and ionization across a sample. In this study, a deuterated internal standard was used to account for these signal variations. Initial experiments were performed using pure standards and optimal cutting temperature compound (OCT) to generate known areas of ion suppression. By monitoring the analyte-to-internal-standard ratio, differences in ion signal were taken into account, resulting in images that better represented the analyte concentration. These experiments were then replicated using multiple tissue types in which the analyte's MS signal was monitored. In certain tissues, including liver and kidney, the analyte signal was attenuated by up to 90%; however, when the analyte-to-internal-standard ratio was monitored, these differences were taken into account. These experiments further exemplify the need for an internal standard in the MSI workflow.


Concurrent robotic renal and prostatic surgery: initial case series and safety data of a new surgical technique.
In the era of prostate-specific antigen screening and frequent cross-sectional abdominal imaging, concurrent prostate cancer and renal masses are being identified and treated. Minimizing patient morbidity and cost by avoiding separate surgical procedures is advantageous, provided technical feasibility, and safety data. Our goal was to assess the feasibility and safety of single-setting robotic renal surgery and prostatectomy. We present our initial experience. To assess the feasibility and safety of single-setting concurrent robot-assisted renal surgery and radical prostatectomy utilizing the same port access scheme. From February 2009 to June 2009, we performed single-setting concurrent robot-assisted radical nephrectomy/partial nephrectomy and radical prostatectomy on two patients with synchronous kidney tumors and prostate cancer. Identical port sites were used during both aspects of the procedure with the exception of one additional port during prostatectomy. Prostate cancer clinical stage and Gleason scores were T1c and 6 and T2a and 7, respectively. Corresponding renal tumors were 5 cm, respectively. Both operations were performed, with no conversion to open surgery. There were no intraoperative complications and the postoperative course was uneventful in both patients. Discharge was on postoperative day 2 and 3, respectively. Patient 2 had an episode of delayed bleeding on postoperative day 9, treated by selective angio-embolization. Mean operative time for nephrectomy and prostatectomy (135 and 139 minutes, respectively) and estimated blood loss (75 and 100 mL, respectively) were reasonable. We began with the renal portion utilizing a lateral decubitus position before re-positioning into the lithotomy position for the prostatic portion. Clamping time was 34 minutes during partial nephrectomy. Single-setting robotic radical/partial nephrectomy and radical prostatectomy is technically feasible and safe in properly selected patients who present with synchronous primary renal and prostate malignancies.


Control of monocyte recruitment by chemotactic factor(s) in lesion-prone areas of swine aorta.
Aortic areas predisposed to early atherosclerosis in swine demarcated by Evans blue uptake (blue areas) show preferential intimal penetration by blood monocytes before and during lesion formation. These cells are thought to be the major source of foam cells in these early lesions. To examine mechanisms controlling monocyte migration into these areas, extracts of aortic tissue from both blue and white areas of hypercholesterolemic (H) and normal (N) swine were tested for chemotactic activity against monocytes from N and H swine and neutrophils from H swine. The data indicate that extracts of blue areas from H swine contained at least one, and possibly two, factors chemotactic only for monocytes from H swine, but not N swine. These factors were not present in similar extracts from white areas of H swine, or either blue or white area extracts from N swine. Gel filtration chromatography of crude blue area extracts separated two chemotactically active fractions of molecular weights 68,000 and 5000 that were not present in white area extracts and that elicited a positive chemotactic effect only for H swine monocytes. These findings provide a mechanism explaining our previous findings of preferential monocyte adhesion and intimal penetration in lesion-prone areas, and indicate that control of monocyte recruitment into the artery involves not only alteration of the arterial wall, but also functional changes in the circulating monocyte.


Chronic pressure overload cardiac hypertrophy and failure in guinea pigs: II. Cytoskeletal remodeling.
The cytoskeleton is a major regulator of cell shape. To explore potential mechanisms for maladaptation of cardiac myocyte shape in pressure overload-induced congestive heart failure, the abundance and organization of major intra- and extra-myofibrillar cytoskeleton of cardiac myocytes were examined with western blotting and confocal microscopy in guinea pigs with chronic aortic stenosis. It was found that: (1) the amount and distribution of alpha-actinin and myomesin remained unchanged at both the compensated hypertrophy and the congestive heart failure stages; (2) loss of titin was associated with myocyte lengthening in heart failure; (3) desmin protein and filaments in LV myocytes increased progressively with mechanical overload cardiac hypertrophy and subsequent heart failure; (4) a newly developed and validated quantitative confocal microscopic approach disclosed that the microtubule density in isolated LV myocytes increased by 21% at 4 weeks and by 48% 6 months after aortic constriction; (5) at the heart failure stage, microtubule density in LV myocytes showed a statistically significant inverse correlation to the LV maximum +dP/dt and a direct correlation to LV myocyte lengthening; (6) the increased microtubule density in LV myocytes in this model was not due to an increase in total tubulin; and (7) microtubule density in left atrial and right ventricular myocytes also increased when they underwent hypertrophy secondary to left heart failure. These results suggest that the down-regulation of titin and up-regulation of microtubule and desmin filaments may contribute to myocyte lengthening and malfunction in pressure overload congestive heart failure.


Evidence for a prospective anti-osteoporosis effect of black tea (Camellia Sinensis) extract in a bilaterally ovariectomized rat model.
The purpose of this study was to examine whether whole aqueous black tea extract (BTE) prevents bone loss induced by ovarian hormone deficiency. Eighteen 95-100 days old female albino rats were randomly assigned to three treatment groups [sham -operated control (sham); bilaterally ovariectomized (ovx) and ovx + aqueous black tea extract (BTE) ] and sacrificed after 28 days. All animals were fed a standard laboratory diet with free access to deionized water except on days of urinary parameter studies when animals were given only calcium free deionized water during the entire 24 h period of urine collection. Body weight study revealed that rats in the ovx group had significantly higher final body weight than rats in the sham group. This higher final body weight was not observed in animals receiving BTE. The ovx group also had significantly higher abdominal fat mass and liver weight and significantly lower uterus, right kidney and left kidney weights than in other two groups. All these organ weight changes in ovx group also were not observed in animals receiving BTE. Results of urinary studies revealed that rats in the ovx group had significantly higher urinary excretion of calcium (Ca), phosphate, creatinine (Cr), calcium to creatinine (Ca:Cr) ratio (P< 0.001) and hydroxyproline (HPr) (P< 0.01) than rats in the sham group. Significant recovery of all these parameters were observed in animals receiving BTE. The ovx group also had significantly higher (P< 0.001) serum alkaline phosphatase (AP) and tartrate-resistant acid phosphatase (TRAP) activity than rats in the other two groups. These changes could not be seen in animals receiving BTE. Also, identical changes were seen in bone density experiments. Rats in the ovx group had significantly lower densities of the right femur (P<0.001), eighth thoracic rib (P< 0.001), eighth thoracic vertebra (P< 0.05), and fourth lumbar vertebra (P< 0.01) than rats in the sham group; and significant improvement in densities of these bones were seen in animals supplemented with BTE. Animals of ovx group also showed significant decrease in calcium and phosphate level in all these bones which could be regained significantly when these animals were supplemented with BTE. Our findings suggest that aqueous BTE may be effective in preventing bone loss due to ovarian hormone deficiency. Because serum activity of AP, TRAP and urinary loss of bone minerals (Ca and Phosphate) and also the organic components of bone (Cr and HPr) were significantly greater in the ovx group, compared to sham animals and ovx + BTE group. This confirms that ovariectomy enhances and BTE suppresses the rate of bone turnover. The density results of ovx + BTE group are significantly greater than rats in the ovx group, suggesting further that formation exceeded resorption. Detailed studies are underway to clarify the mechanism of this protective effect of BTE on hypogonadal bone loss.


Projections of global health outcomes from 2005 to 2060 using the International Futures integrated forecasting model.
To develop an integrated health forecasting model as part of the International Futures (IFs) modelling system. The IFs model begins with the historical relationships between economic and social development and cause-specific mortality used by the Global Burden of Disease project but builds forecasts from endogenous projections of these drivers by incorporating forward linkages from health outcomes back to inputs like population and economic growth. The hybrid IFs system adds alternative structural formulations for causes not well served by regression models and accounts for changes in proximate health risk factors. Forecasts are made to 2100 but findings are reported to 2060. The base model projects that deaths from communicable diseases (CDs) will decline by 50%, whereas deaths from both non-communicable diseases (NCDs) and injuries will more than double. Considerable cross-national convergence in life expectancy will occur. Climate-induced fluctuations in agricultural yield will cause little excess childhood mortality from CDs, although other climate-health pathways were not explored. An optimistic scenario will produce 39 million fewer deaths in 2060 than a pessimistic one. Our forward linkage model suggests that an optimistic scenario would result in a 20% per cent increase in gross domestic product (GDP) per capita, despite one billion additional people. Southern Asia would experience the greatest relative mortality reduction and the largest resulting benefit in per capita GDP. Long-term, integrated health forecasting helps us understand the links between health and other markers of human progress and offers powerful insight into key points of leverage for future improvements.


The role of exosomes in metastasis and progression of melanoma.
The mechanisms of melanoma metastasis have been the subject of extensive research for decades. Improved diagnostic and therapeutic strategies are of increasing importance for the treatment of melanoma due to its high burden of mortality in the advanced stages of the disease. Intercellular communication is a critical event for the progression of cancer. Collective evidence suggests that exosomes, small extracellular membrane vesicles released by the cells, are important facilitators of intercellular communication between the cells and the surrounding environment. Although the emerging field of exosomes is rapidly gaining traction in the scientific community, there is limited knowledge regarding the role of exosomes in melanoma. This review discusses the multifaceted role of melanoma-derived exosomes in promoting the process of metastasis by modulating the invasive and angiogenic capacity of malignant cells. The future implications of exosome research and the therapeutic potential of exosomes are also discussed.


Synchrony, waves, and spatial hierarchies in the spread of influenza.
Quantifying long-range dissemination of infectious diseases is a key issue in their dynamics and control. Here, we use influenza-related mortality data to analyze the between-state progression of interpandemic influenza in the United States over the past 30 years. Outbreaks show hierarchical spatial spread evidenced by higher pairwise synchrony between more populous states. Seasons with higher influenza mortality are associated with higher disease transmission and more rapid spread than are mild ones. The regional spread of infection correlates more closely with rates of movement of people to and from their workplaces (workflows) than with geographical distance. Workflows are described in turn by a gravity model, with a rapid decay of commuting up to around 100 km and a long tail of rare longer range flow. A simple epidemiological model, based on the gravity formulation, captures the observed increase of influenza spatial synchrony with transmissibility; high transmission allows influenza to spread rapidly beyond local spatial constraints.


Telomerase inhibitory effects of medicinal mushrooms and lichens, and their anticancer activity.
Telomerase has been widely accepted as a cancer marker and a promising therapeutic target for novel anticancer drugs. The aim of this study was to investigate the in vitro telomerase inhibitory effects of mushrooms and their anticancer properties. The inhibitory effects of mushrooms and lichens against telomerase activity of HL-60 cells were systematically assessed using polymerase chain reaction based on assay of telomeric repeat amplification protocol. Telomerase inhibitory samples were further tested for antiproliferation effects against the gastric cell line SNU-1 using the MTT method. Ethyl acetate extract of Pleurotus ostreatus, ethyl acetate and water extracts of Lasiosphaera fenzlii, hexane extract of Strobilomyces floccopus, water extract of Sarcodon aspratus, and hexane, ethyl acetate, and water extracts from Umbilicaria esculenta showed strong positive telomerase inhibitory activity. Hexane extract of S. floccopus and water extracts from the edible lichen U. esculenta exhibited strong anticancer effects against SNU-1 cells through antiproliferation assay. The water extract of U. esculenta has a great potential to be developed into an anticancer agent that targets telomerase.


Routine assessment of occupational exposure and its relation to semen quality in infertile men: a cross-sectional study.
Concerns about the detrimental effects of occupational and environmental exposure on male reproductive function have been raised by reports of declining sperm quality over the last decades. The aim of this study was to investigate the association between altered semen parameters and exposure to occupational risk factors as assessed by questionnaire. We conducted a cross-sectional questionnaire-based study among a population of 2122 men who underwent andrological investigation for couple infertility. All participants were interviewed and their semen samples were analyzed. Information about medical history and occupational exposure was used to classify participants into exposed and unexposed groups. Exposure to pesticides was associated with a significantly higher risk of asthenozoospermia (adjusted odds ratio [OR] = 1.6; 95% CI, 1.0-2.4) and necrozoospermia (OR = 2.6; 95% CI, 1.4-4.7). Exposure to cement was found to be correlated with a higher risk of oligozoospermia (OR = 1.1; 95% CI, 0.9-1.4). There was no association between semen impairment and exposure to solvents, excess heat, or mechanical vibrations. We found an association between self-reported occupational exposure and altered semen parameters. These results support the usefulness of questionnaires for routine assessment and management of occupational exposures in infertile men.


Hedgehog signaling plays a cell-autonomous role in maximizing cardiac developmental potential.
Elucidation of the complete roster of signals required for myocardial specification is crucial to the future of cardiac regenerative medicine. Prior studies have implicated the Hedgehog (Hh) signaling pathway in the regulation of multiple aspects of heart development. However, our understanding of the contribution of Hh signaling to the initial specification of myocardial progenitor cells remains incomplete. Here, we show that Hh signaling promotes cardiomyocyte formation in zebrafish. Reduced Hh signaling creates a cardiomyocyte deficit, and increased Hh signaling creates a surplus. Through fate-mapping, we find that Hh signaling is required at early stages to ensure specification of the proper number of myocardial progenitors. Genetic inducible fate mapping in mouse indicates that myocardial progenitors respond directly to Hh signals, and transplantation experiments in zebrafish demonstrate that Hh signaling acts cell autonomously to promote the contribution of cells to the myocardium. Thus, Hh signaling plays an essential early role in defining the optimal number of cardiomyocytes, making it an attractive target for manipulation of multipotent progenitor cells.


Dementia case management through the eyes of informal carers: A national evaluation study.
This paper focuses on the evaluation of dementia case management in the Netherlands, as well as factors associated with positive evaluations of informal caregivers. A survey was completed by 554 informal carers. The majority of the informal carers were older (69% was 55+), and female (73%), and often concerned the partner or adult children of the person with dementia. Eighty percent indicated that the contact with the case manager facilitated their role as informal carer, while 95% or more stated that the case manager showed sufficient understanding, allowed enough space to decide together on how to approach problems in the care, took time to listen to their story, gave sufficient attention to and showed interest in their relative, took their schedule into account and/or kept appointments. Contrary to the expectations, multilevel analyses did not show association between informal caregivers' care burden and the evaluation of case management. Neither were the period living with dementia and the number of personal contacts with the case manager associated with the evaluations of informal caregivers. However, being the partner of the patient was significantly related ( p < 0.05) to a positive overall evaluation by informal carers. These results suggest that sufficient case management resources should be offered and targeted especially towards partners of people with dementia.


New approach to radial expansive force measurement of self expandable esophageal metal stents.
The accurate measurement of radial expansive force is crucial for optimal design and implantation of self expandable esophageal metal stents. In the present study, a new method of measurement under experimental conditions simulating actual stent implantation has been developed. This method offers precise and reproducible measurements and can be applied to a wide variety of stent types. In particular, the method enables one to measure expansive pressure as well as the true radial expansive force up to the radial compression ratio of 72%, covering the range of compression often encountered in a partially obstructed lumen. The test results for various kinds of metal stents are presented and compared. Based on these results, three important points of observation critical in explaining and predicting the expansion characteristics of stents have been reported. Further understanding and characterization of these findings will be necessary for developing new stents with outstanding clinical efficacy.


Reducing blood culture contamination in community hospital emergency departments: a multicenter evaluation of a quality improvement intervention.
Blood culture contamination is a common and preventable problem in the emergency department (ED). In a previous single-center study, changing the process of ED blood culture collection from the traditional "clean," nonsterile procedure to a fully sterile procedure with standardized use of sterile gloves, large-volume chlorhexidine skin antisepsis, and fenestrated sterile drapes resulted in a substantial reduction in contamination. The objective of the current study was to evaluate the effectiveness of this sterile blood culture collection process for reducing blood culture contamination in two community hospital EDs. The authors implemented the sterile blood culture collection process in the ED of two hospitals, including Hospital A, which historically had a contamination rate of approximately 5%, and Hospital B, with a 2.5% historical contamination rate. With an interrupted times-series design and segmented regression analysis to adjust for secular trends and autocorrelation, the monthly percentages of cultures contaminated at each hospital during an intervention period (sterile technique) were compared to a 10-month baseline period immediately preceding implementation (clean technique). At Hospital A, the full sterile blood culture collection process was used throughout the 16-month intervention period. At Hospital B, user feedback indicated poor adherence to the process due to difficulty implementing the fenestrated drape component; therefore, the process was simplified to the modified sterile collection process, in which the fenestrated drape component was dropped and sterile gloves and large-volume skin antisepsis were emphasized. Hence, at Hospital B, two intervention periods were compared to the baseline period: the 8-month intervention period 1 (full sterile process) and the subsequent 8-month intervention period 2 (modified sterile process). At Hospital A, during the baseline period, 165 of 3,417 (4.83%) cultures were contaminated, while 142 of 5,238 (2.71%) were contaminated during the intervention period (p < 0.01). In the segmented regression model, the full sterile blood culture collection process was associated with an immediate 2.68% (95% confidence interval [CI] = 1.43% to 3.52%) absolute reduction in contamination and sustained reductions during the entire intervention period. At Hospital B, during the baseline, 63 of 2,509 (2.51%) cultures were contaminated. In intervention period 1 with the full sterile process, 51 of 1,865 (2.73%) cultures were contaminated (p = 0.65), with segmented regression results showing no changes compared to baseline. After simplification of the process to address poor adherence, the modified sterile process during intervention period 2 was associated with a significant reduction in contamination, with 17 of 1,860 (0.91%) cultures contaminated (p < 0.01 compared to baseline). The segmented regression model demonstrated the modified sterile process was associated with an immediate 1.53% (95% CI = 1.00% to 1.88%) absolute reduction in contamination with significant sustained reductions. Changing the method of blood culture collection from the commonly used nonsterile technique to a sterile process resulted in significant reductions in blood culture contamination at two community hospital EDs, including one with low baseline contamination. Monitoring the implementation process at both sites was important to identify and overcome operational challenges. At one study site, simplification of the process by removing the fenestrated drape component was a key for successful implementation.


Thinness and weight loss: beneficial or detrimental to longevity?
This review examined the hypotheses that 1) low body mass index (BMI) is optimal for longevity and 2) weight loss reduces mortality rates. The preponderance of epidemiological evidence fails to support either of these hypotheses. Indeed, a number of studies show that thinness and weight loss (regardless of initial BMI) are associated with increased mortality rates. These findings cannot be attributed to smoking status or to weight loss resulting from subclinical disease. The effect of intentional weight loss on mortality rates depends upon health status. For overweight individuals in good health, there is no compelling evidence to show that mortality rates are reduced with weight loss. Even among overweight persons with one or more obesity-related health conditions, specific weight loss recommendations may be unnecessary: 1) the reduction in mortality rate associated with intentional weight loss is independent of the amount of weight loss, 2) the reductions in all-cause mortality rate associated with increased physical activity and fitness (23-44%), independent of changes in body weight, are greater than that reported for intentional weight loss (approximately 20%), and 3) many obesity-related health conditions (e.g., hypertension, dyslipidemias, insulin resistance, glucose intolerance) can be ameliorated independently of weight loss. In view of the potential risks associated with weight loss and weight cycling, it is suggested that public health may be better served by placing greater emphasis on lifestyle changes and less attention to weight loss per se.


Musculoskeletal disorders of the lower limb--ultrasound and magnetic resonance imaging correlation.
The purpose of this paper is to familiarize general radiologists and specialists with the sonographic and corresponding magnetic resonance imaging (MRI) appearance of various musculoskeletal disorders of the lower limb. Technologists and radiologists should be familiar with all imaging techniques for the investigation and evaluation of musculoskeletal abnormalities. The role of high-resolution ultrasound (US) is highlighted, as well as the complementary relation between both imaging modalities. We also discuss some of the advantages of US over MRI in the investigation of musculoskeletal disorders of the lower limb. The MRI and US appearances of various articular, periarticular, and soft tissue pathologies of the lower limb are compared and reviewed, and where possible, the advantages of each modality are identified.


Disaster preparedness and the chronic disease needs of vulnerable older adults.
About 80% of older adults have at least one chronic condition that makes them more vulnerable than healthy people during a disaster. These chronic conditions - combined with the physiological, sensory, and cognitive changes experienced as part of aging - result in frail older adults having special needs during emergencies. Planning and coordination among public health and emergency preparedness professionals and professionals who provide services for the aging are essential to meet these special needs. Several tools and strategies already exist to help prepare these professionals to protect and assist older adults during a disaster. These include having professionals from diverse fields work and train in coalitions, ensuring that advocates for older adults participate in community-wide emergency preparedness, and using community mapping data to identify areas where many older adults live.


B7-H4 expression in renal cell carcinoma and tumor vasculature: associations with cancer progression and survival.
B7-H4 is a recently described B7 family coregulatory ligand that has been implicated as an inhibitor of T cell-mediated immunity. Although expression of B7-H4 is typically limited to lymphoid cells, aberrant B7-H4 expression has also been reported in several human malignancies. To date, associations of B7-H4 with clinical outcomes for cancer patients are lacking. Therefore, we examined B7-H4 expression in fresh-frozen tumor specimens from 259 renal cell carcinoma (RCC) patients treated with nephrectomy between 2000 and 2003 and performed correlative outcome analyses. We report that 153 (59.1%) RCC tumor specimens exhibited B7-H4 staining and that tumor cell B7-H4 expression was associated with adverse clinical and pathologic features, including constitutional symptoms, tumor necrosis, and advanced tumor size, stage, and grade. Patients with tumors expressing B7-H4 were also three times more likely to die from RCC compared with patients lacking B7-H4 (risk ratio = 3.05; 95% confidence interval = 1.51-6.14; P = 0.002). Additionally, 211 (81.5%) specimens exhibited tumor vasculature endothelial B7-H4 expression, whereas only 6.5% of normal adjacent renal tissue vessels exhibited endothelial B7-H4 staining. Based on these findings, we conclude that B7-H4 has the potential to be a useful prognostic marker for patients with RCC. In addition, B7-H4 represents a target for attacking tumor cells as well as tumor neovasculature to facilitate immunotherapeutic treatment of RCC tumors. Last, we demonstrate that patients with RCC tumors expressing both B7-H4 and B7-H1 are at an even greater risk of death from RCC.


Semuloparin for prevention of venous thromboembolism after major orthopedic surgery: results from three randomized clinical trials, SAVE-HIP1, SAVE-HIP2 and SAVE-KNEE.
Semuloparin is a novel ultra-low-molecular-weight heparin under development for venous thromboembolism (VTE) prevention in patients at increased risk, such as surgical and cancer patients. Three Phase III studies compared semuloparin and enoxaparin after major orthopedic surgery: elective knee replacement (SAVE-KNEE), elective hip replacement (SAVE-HIP1) and hip fracture surgery (SAVE-HIP2). All studies were multinational, randomized and double-blind. Semuloparin and enoxaparin were administered for 7-10 days after surgery. Mandatory bilateral venography was to be performed between days 7 and 11. The primary efficacy endpoint was a composite of any deep vein thrombosis, non-fatal pulmonary embolism or all-cause death. Safety outcomes included major bleeding, clinically relevant non-major (CRNM) bleeding, and any clinically relevant bleeding (major bleeding plus CRNM). In total, 1150, 2326 and 1003 patients were randomized in SAVE-KNEE, SAVE-HIP1 and SAVE-HIP2, respectively. In all studies, the incidences of the primary efficacy endpoint were numerically lower in the semuloparin group vs. the enoxaparin group, but the difference was statistically significant only in SAVE-HIP1. In SAVE-HIP1, clinically relevant bleeding and major bleeding were significantly lower in the semuloparin vs. the enoxaparin group. In SAVE-KNEE and SAVE-HIP2, clinically relevant bleeding tended to be higher in the semuloparin group, but rates of major bleeding were similar in the two groups. Other safety parameters were generally similar between treatment groups. Semuloparin was superior to enoxaparin for VTE prevention after hip replacement surgery, but failed to demonstrate superiority after knee replacement surgery and hip fracture surgery. Semuloparin and enoxaparin exhibited generally similar safety profiles.


Factors Causing Seasonal Forms in Ascia monuste (Lepidoptera).
Light effects will produce the seasonal forms of Ascia monuste L. but they are not yet proved to be the only causative agent. The long-day form is melanic; the short-day form is white. Only females exhibit the effect. The capacity to form melanic females is most frequent in the Florida population and is probably genetically determined.


Evidence for splicing of interrupted immunoglobulin variable and constant region sequences in nuclear RNA.
Evidence is presented that the mouse light-chain coding sequence is interrupted in a 27S nuclear RNA species, whereas the sequence is continuous in both a 13S nuclear RNA and in cytoplasmic mRNA. The discontinuity of coding regions in the 27S nuclear RNA parallels the situation found in myeloma DNA and indicates, therefore, that the removal of interruptions in the V and C regions occurs at the level of nuclear RNA.


Regulation of cell survival by secreted proneurotrophins.
Neurotrophins are growth factors that promote cell survival, differentiation, and cell death. They are synthesized as proforms that can be cleaved intracellularly to release mature, secreted ligands. Although proneurotrophins have been considered inactive precursors, we show here that the proforms of nerve growth factor (NGF) and the proforms of brain derived neurotrophic factor (BDNF) are secreted and cleaved extracellularly by the serine protease plasmin and by selective matrix metalloproteinases (MMPs). ProNGF is a high-affinity ligand for p75(NTR) with high affinity and induced p75NTR-dependent apoptosis in cultured neurons with minimal activation of TrkA-mediated differentiation or survival. The biological action of neurotrophins is thus regulated by proteolytic cleavage, with proforms preferentially activating p75NTR to mediate apoptosis and mature forms activating Trk receptors to promote survival.


Controlled surface functionality of magnetic nanoparticles by layer-by-layer assembled nano-films.
Over the past several years, the preparation of functionalized nanoparticles has been aggressively pursued in order to develop desired structures, compositions, and structural order. Among the various nanoparticles, iron oxide magnetic nanoparticles (MNPs) have shown great promise because the material generated using these MNPs can be used in a variety of biomedical applications and possible bioactive functionalities. In this study, we report the development of various functionalized MNPs (F-MNPs) generated using the layer-by-layer (LbL) self-assembly method. To provide broad functional opportunities, we fabricated F-MNP bio-toolbox by using three different materials: synthetic polymers, natural polymers, and carbon materials. Each of these F-MNPs displays distinct properties, such as enhanced thickness or unique morphologies. In an effort to explore their biomedical applications, we generated basic fibroblast growth factor (bFGF)-loaded F-MNPs. The bFGF-loaded F-MNPs exhibited different release mechanisms and loading amounts, depending on the film material and composition order. Moreover, bFGF-loaded F-MNPs displayed higher biocompatibility and possessed superior proliferation properties than the bare MNPs and pure bFGF, respectively. We conclude that by simply optimizing the building materials and the nanoparticle's film composition, MNPs exhibiting various bioactive properties can be generated.


FISH analysis of selected soft tissue tumors: Diagnostic experience in a tertiary center.
Fluorescence in situ hybridization (FISH) is an important ancillary tool for the classification of bone/soft tissue (BST) tumors. The aim of this study was to evaluate the contribution of FISH to the final classification of common BST entities in the molecular pathology department of the Royal Prince Alfred Hospital (RPAH), which is one of the most important referral centers for the management of sarcomas in Australia. All routine diagnostic FISH tests performed on BST formalin-fixed paraffin embedded (FFPE) tissue specimens at the RPAH in a 5-year period (February, 2010-November, 2015) were reviewed. FISH analyses presented in this study include commercial break-apart probes (SS18, FUS, DDIT3, FUS, USP6, PDGFB, TFE3 and ALK) and a single enumeration (MDM2) probe. There were 434 interpretable FISH assays on BST samples including MDM2 (n=180), SS18 (n=97), FUS (n=64), DDIT3 (n=37), USP6 (n=30), PDGFB (n=13), TFE3 (n=8) and ALK (n=5). Discrepancies between the histopathological diagnosis and the FISH results were seen in 12% of the cases. In this subset of discordant cases, FISH contributed to the re-classification of 7% of cases originally diagnosed as synovial sarcoma (SS18) and 6% of adipocytic neoplasms (MDM2) based on the presence or absence of the expected gene alteration. Our study confirms that paraffin FISH is a sensitive and specific ancillary tool in the diagnosis of BST neoplasms when used in the appropriate clinicopathological context. These findings highlight the need for further ancillary molecular tools in the diagnosis and characterization of challenging cases.


Long-term outcome after spontaneous HBeAg seroconversion in patients with chronic hepatitis B.
During the course of chronic hepatitis B virus (HBV) infection, hepatitis B e antigen (HBeAg) seroconversion to its antibody (anti-HBe) often coincides with normalization of liver biochemical test and clinical remission, but data regarding long-term outcome after spontaneous seroconversion are still scarce. Excluding patients with other virus(es) concurrent infection, 283 patients with chronic HBV infection were followed up for at least 1 year after spontaneous HBeAg seroconversion to anti-HBe. Follow-up studies included clinical, biochemical, and virologic evaluation and hepatocellular carcinoma (HCC) screening with ultrasonography and alpha-fetoprotein assay. During a median follow-up period of 8.6 years (range, 1 to 18.4 years) after HBeAg seroconversion in 283 patients, 189 (66.8%) showed sustained remission, whereas the remaining 94 (33.2%) experienced alanine aminotransferase (ALT) elevation over twice the upper limit of normal: 12 (4.2%) associated with HBeAg reversion, 68 (24%) with detectable serum HBV DNA but HBeAg negative, and 14 (4.9%) of undetermined causes. Of the 269 patients without evidence of cirrhosis at the time of HBeAg seroconversion, 21 (7.8%) developed cirrhosis with a cumulative incidence and relative risk significantly higher in patients developing active hepatitis than in patients with sustained remission (P <.05). HCC developed in 6 (2.2%) of the 283 patients, also with a significantly higher cumulative incidence in patients developing active hepatitis after HBeAg seroconversion (P <.005). In conclusion, the results suggest that spontaneous HBeAg seroconversion confers favorable long-term outcomes. However, active hepatitis still may develop and lead to cirrhosis and HCC.


Attending Physician Adherence to a 29-Component Central Venous Catheter Bundle Checklist During Simulated Procedures.
Central venous catheter insertions may lead to preventable adverse events. Attending physicians' central venous catheter insertion skills are not assessed routinely. We aimed to compare attending physicians' simulated central venous catheterinsertion performance to published competency standards. Prospective cohort study of attending physicians' simulated internal jugular and subclavian central venous catheter insertion skills versus a historical comparison group of residents who participated in simulation training. Fifty-eight Veterans Affairs Medical Centers from February 2014 to December 2014 during a 2-day simulation-based education curriculum and two academic medical centers in Chicago. A total of 108 experienced attending physicians and 143 internal medicine and emergency medicine residents. None. Using a previously published central venous catheter insertion skills checklist, we compared Veterans Affairs Medical Centers attending physicians' simulated central venous catheter insertion performance to the same simulated performance by internal medicine and emergency medicine residents from two academic centers. Attending physician performance was compared to residents' baseline and posttest (after simulation training) performance. Minimum passing scores were set previously by an expert panel. Attending physicians performed higher on the internal jugular (median, 75.86% items correct; interquartile range, 68.97-86.21) and subclavian (median, 83.00%; interquartile range, 59.00-86.21) assessments compared to residents' internal jugular (median, 37.04% items correct; interquartile range, 22.22-68.97) and subclavian (median, 33.33%; interquartile range, 0.00-70.37; both p < 0.001) baseline assessments. Overall simulated performance was poor because only 12 of 67 attending physicians (17.9%) met or exceeded the minimum passing score for internal jugular central venous catheter insertion and only 11 of 47 (23.4%) met or exceeded the minimum passing score for subclavian central venous catheter insertion. Resident posttest performance after simulation training was significantly higher than attending physician performance (internal jugular: median, 96%; interquartile range, 93.10-100.00; subclavian: median, 100%; interquartile range, 96.00-100.00; both p < 0.001). This study demonstrates highly variable simulated central venous catheter insertion performance among a national cohort of experienced attending physicians. Hospitals, healthcare systems, and governing bodies should recognize that even experienced physicians require periodic clinical skill assessment and retraining.


Characterization of neutralizing antibodies reacting with the 213-224 amino-acid segment of human galectin-9.
Extra-cellular galectin-9 (gal-9) is an immuno-modulatory protein with predominant immunosuppressive effects. Inappropriate production of gal-9 has been reported in several human malignancies and viral diseases like nasopharyngeal, pancreatic and renal carcinomas, metastatic melanomas and chronic active viral hepatitis. Therefore therapeutic antibodies neutralizing extra-cellular gal-9 are expected to contribute to immune restoration in these pathological conditions. Two novel monoclonal antibodies targeting gal-9 -Gal-Nab 1 and 2-have been produced and characterized in this study. We report a protective effect of Gal-Nab1 and Gal-Nab2 on the apoptotic cell death induced by gal-9 in primary T cells. In addition, they inhibit late phenotypic changes observed in peripheral T cells that survive gal-9-induced apoptosis. Gal-Nab1 and Gal-Nab2 bind nearly identical, overlapping linear epitopes contained in the 213-224 amino-acid segments of gal-9. Nevertheless, they have some distinct functional characteristics suggesting that their three-dimensional epitopes are distinct. These differences are best demonstrated when gal-9 is applied on Jurkat cells where Gal-Nab1 is less efficient than Gal-Nab2 in the prevention of apoptotic cell death. In addition, Gal-Nab1 stimulates non-lethal phosphatidylserine translocation at the plasma membrane and calcium mobilization triggered by gal-9 in these cells. Both Gal-Nab1 and 2 cross-react with murine gal-9. They bind its natural as well as its recombinant form. This cross-species recognition will be an advantage for their assessment in pre-clinical tumor models.


A generalized quantitative antibody homeostasis model: antigen saturation, natural antibodies and a quantitative antibody network.
In a pair of articles, we present a generalized quantitative model for the homeostatic function of clonal humoral immune system. In this second paper, we describe how antibody production controls the saturation of antigens and the network of antibody interactions that emerges in the epitome space with the establishment of the immune system. Efficient control of antigens, be it self or foreign, requires the maintenance of antibody concentrations that saturate antigen to relevant levels. Simple calculations suggest that the observed diverse recognition of antigens by natural antibodies is only possible by cross-reactivity whereby particular clones of antibodies bind to diverse targets and shared recognition of particular antigens by multiple antibody clones contribute to the maintenance of antigen control. We also argue that natural antibodies are none else than the result of thymus-independent responses against immunological self. We interpret and explain antibody production and function in a virtual molecular interaction space and as a network of interactions. Indeed, the general quantitative (GQM) model we propose is in agreement with earlier models, confirms some assumptions and presumably provides the theoretical basis for the construction of a real antibody network using the sequence and interaction database data.


Serum lipids, proteins and electrolyte profiles in rats following total body irradiation.
Serum lipid and electrolyte imbalances are common in critically ill patients undergoing radiation therapy. Although multiple disease states and medication may be responsible for the development of these disorders, the aim of this research is to sequentially document the effect of total body radiation on body function utilizing the sequential changes in the serum lipids, electrolytes and protein in rats. Serum protein and lipids contents were assessed using kits while electrolytes were assessed with flame photometry in rats exposed to total body irradiations of 1.27 Gy/min in cumulative doses to the fourth irradiation at five-day intervals. Total cholesterol and triacylglycerols serum levels were significantly reduced by irradiation (p < 0.05). No significant differences between experimental and control groups for HDL-C serum levels were detected. Serum electrolyte concentration remained within the normal range after each total body irradiation. Sodium, bicarbonate and chloride were significantly (p < 0.05) higher than control while potassium and creatinine were significantly reduced after the first irradiation only. Sodium/potassium ratio was significantly (p < 0.05) elevated. Serum protein was significantly (p < 0.05) elevated with increasing radiation. There are subtle but significant changes in serum lipids, electrolytes and protein after total body irradiation of normal rats. These variations could be due to non-specific stress reactions; as such, they are important markers in radiation induced injury diagnosis.


Augmentation of mouse liver-associated natural killer activity by biologic response modifiers occurs largely via rapid recruitment of large granular lymphocytes from the bone marrow.
A variety of biologic response modifiers (BRM) can potently augment NK activity in nonlymphoid organs. By using the liver as a model organ, we have shown that this augmentation of organ-associated NK activity is coincident with a 10- to 15-fold increase in the number of large granular lymphocytes (LGL) which can be isolated. The present study was designed to investigate the mechanism by which BRM induce this increase in liver-associated LGL and the coincident increase in hepatic NK activity. Initial studies confirm that a single dose of the pyran copolymer, maleic anhydride divinyl ether (MVE-2), augmented hepatic NK activity and increased the number of liver-associated LGL from 3 x 10(4)/liver to 5 x 10(5)/liver (a 17-fold increase). Multiple injections of MVE-2 further augmented total liver-associated NK activity and LGL number (to 13 x 10(5)/liver). As expected, both the NK activity and detectable LGL were eliminated by treatment of the mice with antiasialo GM1 (asGM1) serum. Three possible mechanisms for the BRM-induced increase in liver-associated LGL have been investigated, including 1) the rapid proliferation of resident hepatic LGL, 2) the redistribution of mature LGL from peripheral sites such as the spleen, or by 3) a rapid output and subsequent hepatic localization of LGL or their precursors recently derived from the bone marrow (BM). Our results demonstrated that the contribution of in situ proliferation to the BRM-induced increase in liver-LGL was relatively small, since the number of cells expressing NK-associated markers (i.e., asGM1, Thy-1.2, and NK1.1) and in G2/M phase (as assessed by propidium iodide uptake) was only 4 to 8%. Further experiments demonstrated that splenectomy before the administration of MVE-2 did not inhibit the augmentation of liver-associated NK activity. This result argued against a recruitment of mature LGL from the spleen. In contrast, selective depletion of the BM following administration of 89Sr decreased the ability of MVE-2 to augment liver-associated NK activity by greater than 80%. This procedure also significantly decreased the ability of Propionibacterium acnes (85%) and multiple doses of IL-2 (49%) to augment liver-associated NK activity. These results demonstrate that the rapid augmentation of liver-associated NK activity by BRM is largely due to localization and accumulation in the liver of LGL recently derived from the BM.


E-cigarettes, Cigarettes, and the Prevalence of Adolescent Tobacco Use.
Adolescent e-cigarette use has increased rapidly in recent years, but it is unclear whether e-cigarettes are merely substituting for cigarettes or whether e-cigarettes are being used by those who would not otherwise have smoked. To understand the role of e-cigarettes in overall tobacco product use, we examine prevalence rates from Southern California adolescents over 2 decades. The Children's Health Study is a longitudinal study of cohorts reaching 12th grade in 1995, 1998, 2001, 2004, and 2014. Cohorts were enrolled from entire classrooms in schools in selected communities and followed prospectively through completion of secondary school. Analyses used data from grades 11 and 12 of each cohort (N = 5490). Among 12th-grade students, the combined adjusted prevalence of current cigarette or e-cigarette use in 2014 was 13.7%. This was substantially greater than the 9.0% adjusted prevalence of current cigarette use in 2004, before e-cigarettes were available (P = .003) and only slightly less than the 14.7% adjusted prevalence of smoking in 2001 (P = .54). Similar patterns were observed for prevalence rates in 11th grade, for rates of ever use, and among both male and female adolescents and both Hispanic and Non-Hispanic White adolescents. Smoking prevalence among Southern California adolescents has declined over 2 decades, but the high prevalence of combined e-cigarette or cigarette use in 2014, compared with historical Southern California smoking prevalence, suggests that e-cigarettes are not merely substituting for cigarettes and indicates that e-cigarette use is occurring in adolescents who would not otherwise have used tobacco products.


Different forms of structural plasticity in the adult olfactory bulb.
The adult olfactory bulb (OB) continuously receives new interneurons that integrate into the functional neuronal network and that play an important role in odor information processing and olfactory behavior. Adult neuronal progenitors are derived from neural stem cells in the subventricular zone (SVZ) bordering the lateral ventricle. They migrate long distances along the rostral migratory stream (RMS) toward the OB where they differentiate into interneurons, mature, and establish synapses with tufted or mitral cells (MC), the principal neurons in the OB. The plasticity provided by both adult-born and pre-existing early-born neurons depends on the formation and pruning of new synaptic contacts that adapt the functioning of the bulbar network to changing environmental conditions. However, the formation of new synapses occurs over a long time scale (hours-days), whereas some changes in environmental conditions can occur more rapidly, requiring a much faster adjustment of neuronal networks. A new form of structural remodeling of adult-born, but not early-born, neurons was recently brought to light. This plasticity, which is based on the activity-dependent relocation of mature spines of GCs toward the dendrites of active principal cells, may allow a more rapid adjustment of the neuronal network in response to quick and persistent changes in sensory inputs. In this mini-review we discuss the different forms of structural plasticity displayed by adult-born and early-born neurons and the possibility that these different forms of structural remodeling may fulfill distinct roles in odor information processing.


Effects of nematicides and plant resistance on white clover performance and seasonal populations of nematodes parasitizing white clover in grazed pasture.
Root-infecting nematodes are a major cause of white clover, Trifolium repens, not reaching its potential in New Zealand pastures. Resistance and/or tolerance are the preferred control options. Greenhouse-based, recurrent selection programs have developed resistance to Meloidogyne trifoliophila and Heterodera trifolii, and a field-based program has developed tolerance. Lines from these programs were compared with commercial cultivars as controls in a series of field trials at four sites over 4 years. Resistant lines from the CCN program performed better than susceptible lines and as well as most cultivars, reflecting the high level of resistance developed in this greenhouse-based program. In stained root from Cambridge, numbers of CCN were lower in resistant lines than in cultivars; numbers in susceptible lines were intermediate. CCN resistance was also reflected to a lesser extent in the number of cysts counted in soil under resistant lines in Palmerston North. The root-knot nematode-resistant material performed better than the susceptible and as well as most cultivars. In one trial of CRKN-resistant lines, resistant and susceptible lines had similar numbers of CRKN which were both lower than the numbers in the cultivars; in the second trial, there were fewer CRKN in resistant than in susceptible lines or cultivars. The tolerant selections, developed under field conditions, performed as well as or better than the cultivars. The selections from the breeding programmes have exhibited strong agronomic potential across locations and years, and the best material has been crossed; progeny are being assessed in current field trials.


Potential toxic effects of glyphosate and its commercial formulations below regulatory limits.
Glyphosate-based herbicides (GlyBH), including Roundup, are the most widely used pesticides worldwide. Their uses have increased exponentially since their introduction on the market. Residue levels in food or water, as well as human exposures, are escalating. We have reviewed the toxic effects of GlyBH measured below regulatory limits by evaluating the published literature and regulatory reports. We reveal a coherent body of evidence indicating that GlyBH could be toxic below the regulatory lowest observed adverse effect level for chronic toxic effects. It includes teratogenic, tumorigenic and hepatorenal effects. They could be explained by endocrine disruption and oxidative stress, causing metabolic alterations, depending on dose and exposure time. Some effects were detected in the range of the recommended acceptable daily intake. Toxic effects of commercial formulations can also be explained by GlyBH adjuvants, which have their own toxicity, but also enhance glyphosate toxicity. These challenge the assumption of safety of GlyBH at the levels at which they contaminate food and the environment, albeit these levels may fall below regulatory thresholds. Neurodevelopmental, reproductive, and transgenerational effects of GlyBH must be revisited, since a growing body of knowledge suggests the predominance of endocrine disrupting mechanisms caused by environmentally relevant levels of exposure.


Correlation between normalized difference vegetation index and malaria in a subtropical rain forest undergoing rapid anthropogenic alteration.
Time-series of coarse-resolution greenness values derived through remote sensing have been used as a surrogate environmental variable to help monitor and predict occurrences of a number of vector-borne and zoonotic diseases, including malaria. Often, relationships between a remotely-sensed index of greenness, e.g. the normalized difference vegetation index (NDVI), and disease occurrence are established using temporal correlation analysis. However, the strength of these correlations can vary depending on type and change of land cover during the period of record as well as inter-annual variations in the climate drivers (precipitation, temperature) that control the NDVI values. In this paper, the correlation between a long (260 months) time-series of monthly disease case rates and NDVI values derived from the Global Inventory Modeling and Mapping Studies (GIMMS) data set were analysed for two departments (administrative units) located in the Atlantic Forest biome of eastern Paraguay. Each of these departments has undergone extensive deforestation during the period of record and our analysis considers the effect on correlation of active versus quiescent periods of case occurrence against a background of changing land cover. Our results show that timeseries data, smoothed using the Fourier Transform tool, showed the best correlation. A moving window analysis suggests that four years is the optimum time frame for correlating these values, and the strength of correlation depends on whether it is an active or a quiescent period. Finally, a spatial analysis of our data shows that areas where land cover has changed, particularly from forest to non-forest, are well correlated with malaria case rates.


Compound prism design principles, I.
Prisms have been needlessly neglected as components used in modern optical design. In optical throughput, stray light, flexibility, and in their ability to be used in direct-view geometry, they excel over gratings. Here we show that even their well-known weak dispersion relative to gratings has been overrated by designing doublet and double Amici direct-vision compound prisms that have 14° and 23° of dispersion across the visible spectrum, equivalent to 800 and 1300 lines/mm gratings. By taking advantage of the multiple degrees of freedom available in a compound prism design, we also show prisms whose angular dispersion shows improved linearity in wavelength. In order to achieve these designs, we exploit the well-behaved nature of prism design space to write customized algorithms that optimize directly in the nonlinear design space. Using these algorithms, we showcase a number of prism designs that illustrate a performance and flexibility that goes beyond what has often been considered possible with prisms.


Confinement and processing effects on glass transition temperature and physical aging in ultrathin polymer films: novel fluorescence measurements.
Fluorescence intensity measurements of chromophore-doped or -labeled polymers have been used for the first time to determine the effects of decreasing film thickness on glass transition temperature, T(g), the relative strength of the glass transition, and the relative rate of physical aging below T(g) in supported, ultrathin polymer films. The temperature dependence of fluorescence intensity measured in the glassy state of thin and ultrathin films of pyrene-doped polystyrene (PS), poly(isobutyl methacrylate) (PiBMA), and poly(2-vinylpyridine) (P2VP) differs from that in the rubbery state with a transition at T(g). Positive deviations from bulk T(g) are observed in ultrathin PiBMA and P2VP films on silica substrates while substantial negative deviations from bulk T(g) are observed in ultrathin PS films on silica substrates. The relative difference in the temperature dependences of fluorescence intensity in the rubbery and glassy states is usually reduced with decreasing film thickness, indicating that the strength of the glass transition is reduced in thinner films. The temperature dependence of fluorescence intensity also provides useful information on effects of processing history as well as on the degree of polymer-substrate interaction. In addition, when used as a polymer label, a mobility-sensitive rotor chromophore is demonstrated to be useful in measuring relative rates of physical aging in films as thin as 10 nm.


Cold-hardiness and evaporative water loss in hatchling turtles.
North American turtles hatch in late summer and spend their first winter either on land or underwater. Adaptations for terrestrial overwintering of hatchlings in northern regions, where winter thermal and hydric regimes are harsh, have not been systematically investigated in many species. We measured intrinsic supercooling capacity, resistance to inoculative freezing, and desiccation resistance in hatchlings of terrestrial and aquatic turtles collected from northern (Terrapene ornata, Chrysemys picta bellii, Kinosternon flavescens, Chelydra serpentina) and southern (Chrysemys picta dorsalis, Trachemys scripta, Sternotherus odoratus, Sternotherus carinatus) locales. Supercooling capacity was estimated from the crystallization temperature of turtles cooled in the absence of external ice nuclei. Mean values ranged from -8.1 degrees to -15.5 degrees C and tended to be lower in terrestrial hibernators. Inoculation resistance was estimated from the crystallization temperature of turtles cooled in a matrix of frozen soil. These values (range of means: -0.8 degrees to -13.6 degrees C) also tended to be lower in the terrestrial hibernators, especially C. picta bellii. Mean rates of evaporative water loss varied markedly among the species (0.9-11.4 mg g(-1) d(-1)) and were lowest in the terrestrial hibernators. Most species tolerated the loss of a modest amount of body water, although half of the sample of S. carinatus died from desiccation. In general, turtles did not regain lost body water from wet soil, and immersion in free water was required for rehydration. Therefore, desiccation resistance may be an important adaptation to terrestrial hibernation. Resistances to inoculative freezing and desiccation were directly correlated, perhaps because they are governed by the same morphological characteristics.


Toward dynamic isotopomer analysis in the rat brain in vivo: automatic quantitation of 13C NMR spectra using LCModel.
The LCModel method was adapted to analyze localized in vivo (13)C NMR spectra obtained from the rat brain in vivo at 9.4 T. Prior knowledge of chemical-shifts, J-coupling constants and J-evolution was included in the analysis. Up to 50 different isotopomer signals corresponding to 10 metabolites were quantified simultaneously in 400 microl volumes in the rat brain in vivo during infusion of [1,6-(13)C(2)]glucose. The analysis remained accurate even at low signal-to-noise ratio of the order of 3:1. The relative distribution of isotopomers in glutamate, glutamine and aspartate determined in vivo in 22 min was in excellent agreement with that measured in brain extracts. Quantitation of time series of (13)C spectra yielded time courses of total (13)C label incorporation into up to 16 carbon positions, as well as time courses of individual isotopomer signals, with a temporal resolution as low as 5 min (dynamic isotopomer analysis). The possibility of measuring in vivo a wealth of information that was hitherto accessible only in extracts is likely to expand the scope of metabolic studies in the intact brain.


Towards complete and error-free genome assemblies of all vertebrate species.
High-quality and complete reference genome assemblies are fundamental for the application of genomics to biology, disease, and biodiversity conservation. However, such assemblies are available for only a few non-microbial species1-4. To address this issue, the international Genome 10K (G10K) consortium5,6 has worked over a five-year period to evaluate and develop cost-effective methods for assembling highly accurate and nearly complete reference genomes. Here we present lessons learned from generating assemblies for 16 species that represent six major vertebrate lineages. We confirm that long-read sequencing technologies are essential for maximizing genome quality, and that unresolved complex repeats and haplotype heterozygosity are major sources of assembly error when not handled correctly. Our assemblies correct substantial errors, add missing sequence in some of the best historical reference genomes, and reveal biological discoveries. These include the identification of many false gene duplications, increases in gene sizes, chromosome rearrangements that are specific to lineages, a repeated independent chromosome breakpoint in bat genomes, and a canonical GC-rich pattern in protein-coding genes and their regulatory regions. Adopting these lessons, we have embarked on the Vertebrate Genomes Project (VGP), an international effort to generate high-quality, complete reference genomes for all of the roughly 70,000 extant vertebrate species and to help to enable a new era of discovery across the life sciences.


Antidiabetic action of somatostatin--assessed by the artificial pancreas.
By means of a glucose-controlled insulin- and glucose-infusion system (GCIGIS) we examined the effect of somatostatin on insulin and glucose requirements following meals or oral glucose loads in juvenile diabetics. In six of seven patients the insulin requirement with somatostatin was remarkably reduced to between 38 per cent and 79 per cent of that of otherwise identical control experiments. No reduction could be found in the seventh case, fed only 575 kcal. In all cases we observed an increase in dextrose demanded from the GCIGIS ranging between 28 per cent and 192 per cent of the control amounts. In addition, a lowering and smoothing of postprandial blood glucose curves caused by somatostatin application was a general finding. It seems to us most likely that the well-known suppression of the secretion of growth hormone and glucagon, both insulin antagonists, is responsible for the antidiabetic action of somatostatin.


Regulation of interleukin 6 receptor expression in human monocytes and monocyte-derived macrophages. Comparison with the expression in human hepatocytes.
IL-6 is a cytokine with pleiotropic biological functions, including induction of the hepatic acute phase response and differentiation of activated B cells into Ig-secreting plasma cells. We found that human peripheral blood monocytes express the IL-6-R, which is undetectable on the large majority of lymphocytes of healthy individuals. Stimulation of monocytes by endotoxin or IL-1 causes a rapid downregulation of IL-6-R mRNA levels and a concomitant enhancement of IL-6 mRNA expression. IL-6 itself was found to suppress the IL-6-R at high concentrations. A gradual decrease of IL-6-R mRNA levels was observed along in vitro maturation of monocytes into macrophages. We show that downregulation of IL-6-R mRNA levels by IL-1 and IL-6 is monocyte specific, since IL-6-R expression is stimulated by both IL-1 and IL-6 in cultured human primary hepatocytes. Our data indicate that under noninflammatory conditions, monocytes may play a role in binding of trace amounts of circulating IL-6. Repression of monocytic IL-6-R and stimulation of hepatocytic IL-6-R synthesis may represent a shift of the IL-6 tissue targets under inflammatory conditions.


Development of Genome-Wide SNP Markers for Barley via Reference- Based RNA-Seq Analysis.
Marker-assisted selection of crop plants requires DNA markers that can distinguish between the closely related strains often used in breeding. The availability of reference genome sequence facilitates the generation of markers, by elucidating the genomic positions of new markers as well as of their neighboring sequences. In 2017, a high quality genome sequence was released for the six-row barley (Hordeum vulgare) cultivar Morex. Here, we developed a de novo RNA-Seq-based genotyping procedure for barley strains used in Japanese breeding programs. Using RNA samples from the seedling shoot, seedling root, and immature flower spike, we mapped next-generation sequencing reads onto the transcribed regions, which correspond to ∼590 Mb of the whole ∼4.8-Gbp reference genome sequence. Using 150 samples from 108 strains, we detected 181,567 SNPs and 45,135 indels located in the 28,939 transcribed regions distributed throughout the Morex genome. We evaluated the quality of this polymorphism detection approach by analyzing 387 RNA-Seq-derived SNPs using amplicon sequencing. More than 85% of the RNA-Seq SNPs were validated using the highly redundant reads from the amplicon sequencing, although half of the indels and multiple-allele loci showed different polymorphisms between the platforms. These results demonstrated that our RNA-Seq-based de novo polymorphism detection system generates genome-wide markers, even in the closely related barley genotypes used in breeding programs.


The liver as a stem cell and lineage system.
We propose that the liver is a stem cell and lineage system with many parallels to lineages in the bone marrow, gut, and epidermis, varying from them only in kinetics. All are organized with three compartments: a slow cycling stem cell compartment with cells expressing a fetal phenotype and responding slowly to injury; an amplification compartment with cells of intermediate phenotype rapidly proliferating in response to regenerative stimuli or acute injuries; and a terminal differentiation compartment in which cells increasingly differentiate and gradually lose their ability to divide. In all systems, both those with slow or rapid kinetics, the various compartments are positioned in a polarized organization, are associated with a gradient in the chemistry of the extracellular matrix, and show lineage-position-dependent growth responses, gene expression, pharmacological and toxicological responses, and reaction to viruses and radiation. In general, known oncogens selectively kill cells in the differentiation compartment inducing chronic regenerative responses of the cells in stem cell and/or amplification compartment. Tumors arise by subsequent transformation of the activated stem cells or early precursor cells. The evidence for a lineage model consists of the data implicating gradients in cell size, ploidy, growth potential, and antigenic and gene expression in the liver parenchyma along the sinusoidal plates. The traditional explanation for this heterogeneity is that it represents adaption of cells to a changing sinusoidal microenvironment dictated by the direction of blood flow. However, we review the extant data and suggest that it more readily supports a lineage model involving a maturation process beginning with stem cells and precursors in the periportal zone and ending with sensescing parenchyma near the central vein. Support for this theory is provided by the studies on phenotypic heterogeneity in liver, investigations into the embryology of the liver, and analyses of the responses of liver to chemical and viral oncogens that induce rapid proliferation of small cells with oval-shaped nuclei, "oval cells," now thought to be closely related to liver stem cells. The lineage model provides clarity and insights into many aspects of liver biology and disease including the limited proliferative ability of in vitro parenchymal cultures, liver regeneration, gene expression, viral infection, hepatocellular carcinogenesis, liver cell transplantation, and aging.


Effects of cassia oil on serum and hepatic uric acid levels in oxonate-induced mice and xanthine dehydrogenase and xanthine oxidase activities in mouse liver.
We investigated the hypouricemic effects of cassia oil extracted from Cinnamomum cassia using hyperuricemic mice induced by potassium oxonate, and its inhibitory actions against liver xanthine dehydrogenase (XDH) and xanthine oxidase (XOD) activities. Oral administration of cassia oil significantly reduced serum and hepatic urate levels in hyperuricemic mice in a time- and dose-dependent manner. At doses of 450 mg/kg of cassia oil or above, serum urate levels of the oxonate-pretreated mice were not different from the normal control mice. Cassia oil at 600 mg/kg was found to be as potent as allopurinol, which reduced hepatic urate levels to lower than normal. In normal mice, urate levels in liver, but not in serum, were altered with dose-dependent decrease after cassia oil treatment. Furthermore, the ratio, liver uric acid/serum uric acid, was determined after cassia oil administration with time- and dose-dependent decreases in hyperuricemic mice. The positive dose-dependent decrease ratio was also observed after cassia oil treatment in the normal animals. The decreased extent of ratio elicited by cassia oil in normal mice appeared to be greater than that in the hyperuricemic animal. In addition, cassia oil significantly exhibited marked reductions in liver XDH/XOD activities, with an apparent dose-dependence in the normal and hyperuricemic mice. The onset of inhibition in enzyme activities elicited by allopurinol was much higher than that elicited by cassia oil. These results suggested that hypouricemic effects of cassia oil could be explained, at least partly, by inhibiting liver in vivo activities of XDH/XOD.


Compensating control participants when the intervention is of significant value: experience in Guatemala, India, Peru and Rwanda.
The Household Air Pollution Intervention Network (HAPIN) trial is a randomised controlled trial in Guatemala, India, Peru and Rwanda to assess the health impact of a clean cooking intervention in households using solid biomass for cooking. The HAPIN intervention-a liquefied petroleum gas (LPG) stove and 18-month supply of LPG-has significant value in these communities, irrespective of potential health benefits. For control households, it was necessary to develop a compensation strategy that would be comparable across four settings and would address concerns about differential loss to follow-up, fairness and potential effects on household economics. Each site developed slightly different, contextually appropriate compensation packages by combining a set of uniform principles with local community input. In Guatemala, control compensation consists of coupons equivalent to the LPG stove's value that can be redeemed for the participant's choice of household items, which could include an LPG stove. In Peru, control households receive several small items during the trial, plus the intervention stove and 1 month of fuel at the trial's conclusion. Rwandan participants are given small items during the trial and a choice of a solar kit, LPG stove and four fuel refills, or cash equivalent at the end. India is the only setting in which control participants receive the intervention (LPG stove and 18 months of fuel) at the trial's end while also being compensated for their time during the trial, in accordance with local ethics committee requirements. The approaches presented here could inform compensation strategy development in future multi-country trials.


Zebrafish as a model for monocarboxyl transporter 8-deficiency.
Allan-Herndon-Dudley syndrome (AHDS) is a severe psychomotor retardation characterized by neurological impairment and abnormal thyroid hormone (TH) levels. Mutations in the TH transporter, monocarboxylate transporter 8 (MCT8), are associated with AHDS. MCT8 knock-out mice exhibit impaired TH levels; however, they lack neurological defects. Here, the zebrafish mct8 gene and promoter were isolated, and mct8 promoter-driven transgenic lines were used to show that, similar to humans, mct8 is primarily expressed in the nervous and vascular systems. Morpholino-based knockdown and rescue experiments revealed that MCT8 is strictly required for neural development in the brain and spinal cord. This study shows that MCT8 is a crucial regulator during embryonic development and establishes the first vertebrate model for MCT8 deficiency that exhibits a neurological phenotype.


Local food environment interventions to improve healthy food choice in adults: a systematic review and realist synthesis protocol.
Local food environments have been linked with dietary intake and obesity in adults. However, overall evidence remains mixed with calls for increased theoretical and conceptual clarity related to how availability of neighbourhood food outlets, and within-outlet food options, influence food purchasing and consumption. The purpose of this work is to develop a programme theory of food availability, supported by empirical evidence from a range of local food environment interventions. A systematic search of the literature will be followed by duplicate screening and quality assessment (using the Effective Public Health Practice Project tool). Realist synthesis will then be conducted according to the Realist And Meta-narrative Evidence Syntheses: Evolving Standards (RAMESES) publication standards, including transparent appraisal, synthesis and drawing conclusions via consensus. The final synthesis will propose an evidence-based programme theory of food availability, including evidence mapping to demonstrate contextual factors, pathways of influence and potential mechanisms. With the paucity of empirically supported programme theories used in current local food environment interventions to improve food availability, this synthesis may be used to understand how and why interventions work, and thus inform the development of theory-driven, evidence-based interventions to improve healthy food choice and future empirical work. PROSPERO CRD42014009808.


Vascular Ehlers-Danlos syndrome: imaging findings.
Vascular Ehlers-Danlos syndrome (EDS), formerly known as EDS type IV, is an autosomal dominant disorder characterized by fragility of medium and large arteries due to type III procollagen deficiency. Our purpose was to review the imaging findings in a cohort of patients with a diagnosis of vascular EDS. The radiologic, surgical, and genetic databases at a single multispecialty medical practice were reviewed for a 35-year period between 1971 and 2006. Thirty-three patients with a clinical diagnosis of vascular EDS were identified. Imaging studies were available for 28 patients, 13 men and 15 women, with a mean age of 39.8 +/- 16 years at the time of diagnosis. A vascular radiologist reviewed a total of 189 imaging examinations: 87 CT, 27 MRI, 59 sonography, and 16 angiography. Vascular abnormalities were present in 22 (78%) of 28 patients. Arterial abnormalities included 41 aneurysms, 19 dissections, 12 ectasias, and 10 occlusions. There was one splenic vein aneurysm and one carotid cavernous fistula. Six patients had a total of 10 parenchymal infarcts involving the brain (n = 5), kidney (n = 3), and spleen (n = 2). Nine patients had 10 hemorrhagic events, five related to spontaneous vascular rupture and five associated with interventional or surgical procedures. Six patients had 13 nonvascular findings. The most common findings were arterial aneurysms and dissections, followed by arterial ectasias and occlusions. Life-threatening complications included hemorrhage and infarcts.


Dying for the weekend: a retrospective cohort study on the association between day of hospital presentation and the quality and safety of stroke care.
To examine the association between day of admission and measures of the quality and safety of the care received by patients with stroke. Retrospective cohort study of patients admitted to hospitals with stroke (codes I60-I64 from the International Statistical Classification of Diseases and Related Health Problems, Tenth Version) from April 1, 2009, through March 31, 2010. English National Health Service public hospitals. PATIENTS during the study period accounted for 93 621 admissions. We used logistic regression to adjust the outcome measures for case mix. Quality and safety measurements using 6 indicators spanning the hospital care pathway, from timely brain scans to emergency readmissions after discharge. Performance across 5 of the 6 measures was significantly lower on weekends (confidence level, 99%). One of the largest disparities was seen in rates of same-day brain scans, which were 43.1% on weekends compared with 47.6% on weekdays (unadjusted odds ratio, 0.83 [95% CI, 0.81-0.86]). In particular, the rate of 7-day in-hospital mortality for Sunday admissions was 11.0% (adjusted odds ratio, 1.26 [95% CI, 1.16-1.37], with Monday used as a reference) compared with a mean of 8.9% for weekday admissions. Strong evidence suggests that, nationally, stroke patients admitted on weekends are less likely to receive urgent treatments and have worse outcomes across a range of indicators. Although we adjusted the results for case mix, we cannot rule out some of the effect being due to unmeasured differences in patients admitted on weekends compared with weekdays. The findings suggest that approximately 350 in-hospital deaths each year within 7 days are potentially avoidable, and an additional 650 people could be discharged to their usual place of residence within 56 days if the performance seen on weekdays was replicated on weekends.


Metabolic effects of acute thiamine depletion are reversed by rapamycin in breast and leukemia cells.
Thiamine-dependent enzymes (TDEs) control metabolic pathways that are frequently altered in cancer and therefore present cancer-relevant targets. We have previously shown that the recombinant enzyme thiaminase cleaves and depletes intracellular thiamine, has growth inhibitory activity against leukemia and breast cancer cell lines, and that its growth inhibitory effects were reversed in leukemia cell lines by rapamycin. Now, we first show further evidence of thiaminase therapeutic potential by demonstrating its activity against breast and leukemia xenografts, and against a primary leukemia xenograft. We therefore further explored the metabolic effects of thiaminase in combination with rapamycin in leukemia and breast cell lines. Thiaminase decreased oxygen consumption rate and increased extracellular acidification rate, consistent with the inhibitory effect of acute thiamine depletion on the activity of the TDEs pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase complexes; these effects were reversed by rapamycin. Metabolomic studies demonstrated intracellular thiamine depletion and the presence of the thiazole cleavage product in thiaminase-treated cells, providing validation of the experimental procedures. Accumulation of ribose and ribulose in both cell lines support the thiaminase-mediated suppression of the TDE transketolase. Interestingly, thiaminase suppression of another TDE, branched chain amino ketoacid dehydrogenase (BCKDH), showed very different patterns in the two cell lines: in RS4 leukemia cells it led to an increase in BCKDH substrates, and in MCF-7 breast cancer cells it led to a decrease in BCKDH products. Immunoblot analyses showed corresponding differences in expression of BCKDH pathway enzymes, and partial protection of thiaminase growth inhibition by gabapentin indicated that BCKDH inhibition may be a mechanism of thiaminase-mediated toxicity. Surprisingly, most of thiaminase-mediated metabolomic effects were also reversed by rapamycin. Thus, these studies demonstrate that acute intracellular thiamine depletion by recombinant thiaminase results in metabolic changes in thiamine-dependent metabolism, and demonstrate a previously unrecognized role of mTOR signaling in the regulation of thiamine-dependent metabolism.


5-HT7 Receptor Contributes to Proliferation, Migration and Invasion in NSCLC Cells.
Because only a small portion of NSCLC (non-small-cell lung cancer) patients benefit from molecular targeted therapy or immunotherapy and do not develop therapeutic resistance, continued research on new targets is warranted. Serotonin has recently emerged as a growth factor for tumor cells, and its receptors may be potential therapeutic targets. The mechanism related to the behavior of the 5-HT7 receptor in NSCLC remains unknown. Both gene expression analysis and immunohistochemical analysis were conducted to evaluate 5-HT7 receptor expression in NSCLC tissues. The correlation between 5-HT7 receptor expression and clinicopathological features was also examined. Cell proliferation was measured using a CCK8 (Cell Counting Kit-8) assay and colony formation, migration and invasion were evaluated by the Transwell assay. siRNA transfection and stimulation with the selective agonist LP211 were used to identify the involvement of molecules in proliferation, migration and invasion. Quantitative real-time chain reaction (qRT-PCR) and Western blotting were used to quantifiy mRNA and protein levels, respectively. Pathway inhibitors facilitated the exploration of possible signaling pathways regulated by the 5-HT7 receptor in migration and invasion. The 5-HT7 receptor was overexpressed in NSCLC tumor tissues compared with adjacent normal lung tissues. High 5-HT7 receptor expression levels were correlated with lymph node metastasis (P=0.007) and advanced TNM stage (P=0.000) in NSCLC patients. The 5-HT7 receptor positively regulated cell proliferation, migration and invasion in NSCLC cells. The stimulatory effect of the 5-HT7 receptor on A549 cell migration and invasion may occur through the P38 pathway. In H1299 cells, the 5-HT7 receptor might positively regulate Src to promote cell migration and invasion. Our findings suggest that the 5-HT7 receptor, which mediates NSCLC progression, may be a potential therapeutic target.


High-Fructose Diet-Induced Metabolic Disorders Were Counteracted by the Intake of Fruit and Leaves of Sweet Cherry in Wistar Rats.
Numerous studies have indicated that the use of plants rich in bioactive compounds may reduce the risk of non-communicable diseases. The aim of this study was to investigate how the addition of fruit and leaves to high-fructose diet affects lipid metabolism, including the expression of genes involved in fatty acid synthesis and oxidation in the liver and adipose tissue, as well as oxidative stress and inflammation in Wistar rats. The animals were fed with AIN-93G diet, high fructose (HFr) diet, HFr diet with addition of 5% or 10% freeze-dried fruits, and HFr diet with addition of 1% or 3% freeze-dried leaves. The experiment lasted 12 weeks. The results showed that the intake of fruit and leaves of sweet cherry caused the improvement of the liver function, as well as beneficially affected lipid metabolism, among others, by regulating the expression of genes associated with fatty acid synthesis and β-oxidation. Additionally, they exhibited antioxidant and anti-inflammatory properties. In conclusion, the addition of fruit and leaves reduced the adverse changes arising from the consumption of high fructose diet. Therefore, not only commonly consumed fruits, but also leaves can be potentially used as functional foods. These findings may be helpful in prevention and treatment of the obesity-related metabolic diseases, especially cardiovascular diseases.


Quorum sensing triggers the stochastic escape of individual cells from Pseudomonas putida biofilms.
The term 'quorum sensing' (QS) is generally used to describe the phenomenon that bacteria release and perceive signal molecules to coordinate cooperative behaviour in response to their population size. QS-based communication has therefore been considered a social trait. Here we show that QS signals (N-acyl-homoserine lactones, AHLs) are stochastically produced in young biofilms of Pseudomonas putida and act mainly as self-regulatory signals rather than inducing neighbouring cells. We demonstrate that QS induces the expression of putisolvin biosurfactants that are not public goods, thereby triggering asocial motility of induced cells out of microcolonies. Phenotypic heterogeneity is most prominent in the early stages of biofilm development, whereas at later stages behaviour patterns across cells become more synchronized. Our findings broaden our perspective on QS by showing that AHLs can control the expression of asocial (self-directed) traits, and that heterogeneity in QS can serve as a mechanism to drive phenotypic heterogeneity in self-directed behaviour.