Facilitated acquisition of the classically conditioned eyeblink response in active duty military expressing posttraumatic stress disorder symptoms.
Learning diathesis models underscore the role of hyper-conditioning in the development of PTSD following trauma exposure. Eyeblink classical conditioning is one method of examining associative learning biases independent of fear and threat produced by trauma. Facilitated learning is apparent in individuals self-expressing PTSD symptoms, as well as behaviorally inhibited (BI) temperament, a vulnerability factor for PTSD. Here, we examine eyeblink conditioning in active duty military personnel and relate learning with PTSD symptomology. Volunteers were 83 active duty United States Coast Guard personnel (18 females) recruited from small boat stations. Personnel were administered the PTSD Checklist (PCL) to assess current PTSD symptoms using DSM-IV criteria. BI temperament was assessed with the Adult Measure of Behavioural Inhibition (AMBI). Eyeblink conditioning was conducted using a partial reinforcement schedule, whereby paired trials (500-ms pure tone conditioned stimulus co-terminating with a 50-ms air-puff unconditional stimulus) were interpolated with 50% CS-alone trials. Consistent with previous work, there was a high degree of concordance between BI and incidence of PTSD. Further, PTSD was associated with faster learning during the acquisition period, with conditioned responding sustained through the extinction period. These results reinforce the relationship between BI and PTSD in an active duty military sample, supporting previous observations in veteran and civilian samples. The conditioning data are consistent with predictions derived from a learning diathesis model of stress and anxiety, suggesting facilitated associative learning may represent an additional vulnerability for the development and maintenance of stress-related pathology.


The design of schistosomiasis monitoring and evaluation programmes: The importance of collecting adult data to inform treatment strategies for Schistosoma mansoni.
Monitoring and evaluation (M&E) programmes are used to collect data which are required to assess the impact of current interventions on their progress towards achieving the World Health Organization (WHO) goals of morbidity control and elimination as a public health problem for schistosomiasis. Prevalence and intensity of infection data are typically collected from school-aged children (SAC) as they are relatively easy to sample and are thought to be most likely to be infected by schistosome parasites. However, adults are also likely to be infected. We use three different age-intensity profiles of infection for Schistosoma mansoni with low, moderate and high burdens of infection in adults to investigate how the age distribution of infection impacts the mathematical model generated recommendations of the preventive chemotherapy coverage levels required to achieve the WHO goals. We find that for moderate prevalence regions, regardless of the burden of infection in adults, treating SAC only may achieve the WHO goals. However, for high prevalence regions with a high burden of infection in adults, adult treatment is required to meet the WHO goals. Hence, we show that the optimal treatment strategy for a defined region requires consideration of the burden of infection in adults as it cannot be based solely on the prevalence of infection in SAC. Although past epidemiological data have informed mathematical models for the transmission and control of schistosome infections, more accurate and detailed data are required from M&E programmes to accurately determine the optimal treatment strategy for a defined region. We highlight the importance of collecting prevalence and intensity of infection data from a broader age-range, specifically the inclusion of adult data at baseline (prior to treatment) and throughout the treatment programme if possible, rather than SAC only, to accurately determine the treatment strategy for a defined region. Furthermore, we discuss additional epidemiological data, such as individual longitudinal adherence to treatment, that should ideally be collected in M&E programmes.


The importance of pilot studies.
The term 'pilot studies' refers to mini versions of a full-scale study (also called 'feasibility' studies), as well as the specific pre-testing of a particular research instrument such as a questionnaire or interview schedule. Pilot studies are a crucial element of a good study design. Conducting a pilot study does not guarantee success in the main study, but it does increase the likelihood of success. Pilot studies fulfill a range of important functions and can provide valuable insights for other researchers. There is a need for more discussion among researchers of both the process and outcomes of pilot studies.


Identifying Risk Factors for Pressure Injury in Adult Critical Care Patients.
Critically ill patients have a variety of unique risk factors for pressure injury. Identification of these risk factors is essential to prevent pressure injury in this population. To identify factors predicting the development of pressure injury in critical care patients using a large data set from the PhysioNet MIMIC-III (Medical Information Mart for Intensive Care) clinical database. Data for 1460 patients were extracted from the database. Variables that were significant in bivariate analyses were used in a final logistic regression model. A final set of significant variables from the logistic regression was used to develop a decision tree model. In regression analysis, cardiovascular disease, peripheral vascular disease, pneumonia or influenza, cardiovascular surgery, hemodialysis, norepinephrine administration, hypotension, septic shock, moderate to severe malnutrition, sex, age, and Braden Scale score on admission to the intensive care unit were all predictive of pressure injury. Decision tree analysis revealed that patients who received norepinephrine, were older than 65 years, had a length of stay of 10 days or less, and had a Braden Scale score of 15 or less had a 63.6% risk of pressure injury. Determining pressure injury risk in critically ill patients is complex and challenging. One common pathophysiological factor is impaired tissue oxygenation and perfusion, which may be nonmodifiable. Improved risk quantification is needed and may be realized in the near future by leveraging the clinical information available in the electronic medical record through the power of predictive analytics.


Spatial processes decouple management from objectives in a heterogeneous landscape: predator control as a case study.
Predator control is often implemented with the intent of disrupting top-down regulation in sensitive prey populations. However, ambiguity surrounding the efficacy of predator management, as well as the strength of top-down effects of predators in general, is often exacerbated by the spatially implicit analytical approaches used in assessing data with explicit spatial structure. Here, we highlight the importance of considering spatial context in the case of a predator control study in south-central Utah. We assessed the spatial match between aerial removal risk in coyotes (Canis latrans) and mule deer (Odocoileus hemionus) resource selection during parturition using a spatially explicit, multi-level Bayesian model. With our model, we were able to evaluate spatial congruence between management action (i.e., coyote removal) and objective (i.e., parturient deer site selection) at two distinct scales: the level of the management unit and the individual coyote removal. In the case of the former, our results indicated substantial spatial heterogeneity in expected congruence between removal risk and parturient deer site selection across large areas, and is a reflection of logistical constraints acting on the management strategy and differences in space use between the two species. At the level of the individual removal, we demonstrated that the potential management benefits of a removed coyote were highly variable across all individuals removed and in many cases, spatially distinct from parturient deer resource selection. Our methods and results provide a means of evaluating where we might anticipate an impact of predator control, while emphasizing the need to weight individual removals based on spatial proximity to management objectives in any assessment of large-scale predator control. Although we highlight the importance of spatial context in assessments of predator control strategy, we believe our methods are readily generalizable in any management or large-scale experimental framework where spatial context is likely an important driver of outcomes.


Revision arthroscopic rotator cuff repair: repair integrity and clinical outcome.
Literature regarding the outcomes of revision rotator cuff repair is limited. The purposes of the present study were to report the tendon repair integrity and clinical outcomes for a cohort of patients following revision arthroscopic rotator cuff repair and to examine factors related to tendon healing and the influence of healing on clinical outcomes. Twenty-one of twenty-nine consecutive revision arthroscopic rotator cuff repairs with a minimum of two years of postoperative follow-up were retrospectively reviewed. Outcomes were evaluated on the basis of a visual analog pain scale, the range of motion of the shoulder, the Simple Shoulder Test, the American Shoulder and Elbow Surgeons score, and the Constant score. Ultrasonography was used to examine repair integrity at a minimum of one year following surgery. Ten shoulders underwent arthroscopic repair of a recurrent single-tendon posterior rotator cuff tear, whereas eleven shoulders had repair of both the supraspinatus and infraspinatus. The mean age of the twenty-one subjects was 55.6 years; thirteen subjects were male and eight were female. Complete preoperative and postoperative clinical data were available for nineteen subjects after an average duration of follow-up of thirty-three months. Significant improvements were seen in terms of postoperative pain (p < 0.05), the Simple Shoulder Test score (p < 0.05), the American Shoulder and Elbow Surgeons function (p < 0.05) and total scores (p < 0.05), active forward elevation (p < 0.05), and active external rotation (p < 0.05). Postoperative ultrasound data were available for all twenty-one shoulders after a mean duration of follow-up of twenty-five months. Ten (48%) of the twenty-one shoulders had an intact repair. Seven (70%) of the ten single-tendon repairs were intact, compared with three (27%) of the eleven supraspinatus/infraspinatus repairs (p = 0.05). Patient age (p < 0.05) and the number of torn tendons (p = 0.05) had significant effects on postoperative tendon repair integrity. Shoulders with an intact repair had better postoperative Constant scores (p < 0.05) and scapular plane elevation strength (p < 0.05) in comparison with those with a recurrent tear. Revision arthroscopic rotator cuff repair results in reliable pain relief and improvement in shoulder function in selected cases. Approximately half of the revision repairs can be expected to be intact at a minimum of one year following surgery. Patient age and the number of torn tendons are related to postoperative tendon integrity. The postoperative integrity of the rotator cuff can have a significant influence on shoulder abduction strength and the Constant score.


Characterization of the key aroma compounds in beef and pork vegetable gravies á la chef by application of the aroma extract dilution analysis.
By application of the aroma extract dilution analysis (AEDA) on an aroma distillate isolated from a freshly prepared, stewed beef/vegetable gravy, 52 odor-active compounds were detected in the flavor dilution (FD) factor range of 4-4096. On the basis of high FD factors in combination with the results of the identification experiments, 3-(methylthio)propanal (cooked potato), 3-mercapto-2-methylpentan-1-ol (gravy-like), (E,E)-2,4-decadienal (deep-fried, fatty), 3-hydroxy-4,5-dimethyl-2(5H)-furanone (lovage-like), vanillin (vanilla-like), (E,E)-2,4-nonadienal (deep-fried), and (E)-2-undecenal (metallic) are suggested as key contributors to the aroma of the gravy. To get an insight into the role of the vegetables as sources of gravy odorants, a beef gravy was prepared without vegetables. The AEDA results revealed that, in particular, onions and leek are important sources of gravy aroma compounds, adding particularly the very potent, gravy-like smelling 3-mercapto-2-methylpentan-1-ol to the overall aroma profile. Further compounds that were clearly derived from the vegetables and, thus, are important modifiers of the overall aroma were 4-vinyl-2-methoxyphenol, (E)-beta-damascenone, beta-ionone, 2-isopropyl-3-methoxypyrazine, and 2-(sec-butyl)-3-methoxypyrazine. Interestingly, none of the key odorants detected in the gravy can be assumed to be formed from a reaction between beef and vegetable constituents. A comparison of the odorants in the beef/vegetable gravy with a gravy prepared according to the same procedure, but substituting beef by pork meat, indicated that most of the aroma compounds were identical-although different in FD factors-but the tallowy smelling 12-methyltridecanal was detected as key odorant only in the beef/vegetable gravy.


Identification of distinct specificity determinants in resistance protein Cf-4 allows construction of a Cf-9 mutant that confers recognition of avirulence protein Avr4.
The tomato resistance genes Cf-4 and Cf-9 confer specific, hypersensitive response-associated recognition of Cladosporium carrying the avirulence genes Avr4 and Avr9, respectively. Cf-4 and Cf-9 encode type I transmembrane proteins with extracellular leucine-rich repeats (LRRs). Compared with Cf-9, Cf-4 lacks two LRRs and differs in 78 amino acid residues. To investigate the relevance of these differences for specificity, we exchanged domains between Cf-4 and Cf-9, and mutant constructs were tested for mediating the hypersensitive response by transient coexpression with either Avr4 or Avr9. We show that the number of LRRs is essential for both Cf-4 and Cf-9 function. In addition, Cf-9 specificity resides entirely in the LRR domain and appears to be distributed over several distant LRRs. In contrast, Cf-4 specificity determinants reside in the N-terminal LRR-flanking domain and three amino acid residues in LRRs 13, 14, and 16. These residues are present at putative solvent-exposed positions, and all are required for full Cf-4 function. Finally, we show that Cf-9 carrying the specificity determinants of Cf-4 has recognitional specificity for AVR4. The data indicate that diversifying selection of solvent-exposed residues has been a more important factor in the generation of Cf-4 specificity than has sequence exchange between Cf-4 progenitor genes. The fact that most variant residues in Cf-4 are not essential for Cf-4 specificity indicates that the diverse decoration of R proteins is not fully adapted to confer recognition of a certain avirulence determinant but likely provides a basis for a versatile, adaptive recognition system.


Parental determinants of neonatal body composition.
The prevalence of both childhood and adult obesity is rising in the developed world, and there is increasing interest in its underlying causes. A number of studies suggest a positive relationship between birth weight and childhood body mass index, but less is known about specific prenatal environmental influences on more direct measures of obesity. We used data from the Southampton Women's Survey to investigate parental influences on neonatal body composition ascertained by dual x-ray absorptiometry. Participating mothers were characterized in detail (anthropometry, lifestyle, diet) before and during pregnancy; information was also obtained on their partners. The offspring underwent assessment of fat and lean body mass by dual x-ray absorptiometry within 2 wk of birth. Linear regression methods were used to explore the parental determinants of neonatal body composition. Complete data were available for 448 mother-offspring pairs. Taller women and those with higher parity had offspring with increased birth weight, fat, and lean mass (P < 0.05). Mothers who were taller, of greater parity, had greater fat stores, or walked more slowly also had offspring with greater proportionate body fat at birth (all P < 0.05). There was a weaker trend toward lower percentage fat and greater percentage lean in the offspring of mothers who smoked during pregnancy. Maternal size, parity, smoking history, walking speed, and fat stores are independent determinants of neonatal body composition. If these influences are shown to have persisting effects on body composition through to adulthood, they point to novel public health interventions early in life to prevent later obesity.


A Mobile Multimedia Reminiscence Therapy Application to Reduce Behavioral and Psychological Symptoms in Persons with Alzheimer's.
The goal of this project is to develop a novel and innovative mobile solution to address behavioral and psychological symptoms of dementia (BPSD) that occur in individuals with Alzheimer's. BPSD can include agitation, restlessness, aggression, apathy, obsessive-compulsive and repetitive behaviors, hallucinations, delusions, paranoia, and wandering. Alzheimer's currently affects 5.4 million adults in the United States and that number is projected to increase to 14 million by 2050. Almost 90% of all affected with AD experience BPSD, resulting in increased healthcare costs, heavier burden on caregivers, poor patient outcomes, early nursing home placement, long-term hospitalizations, and misuse of medications. Pharmacological support may have undesirable side effects such as sedation. Nonpharmacological interventions are alternative solutions that have shown to be effective without undesirable side effects. Music therapy has been found to lower BPSD symptoms significantly. Our study is based on combination of the reminiscence and the music therapies where past memorable events are recalled using prompts such as photos, videos, and music. We are proposing a mobile multimedia solution, a technical version of the combined reminiscence, and music therapies to prevent the occurrence of BPSD, especially for the rural population who have reduced access to dementia care services.


Lutropin-choriogonadotropin receptor: an unusual member of the G protein-coupled receptor family.
A complementary DNA (cDNA) for the rat luteal lutropin-choriogonadotropin receptor (LH-CG-R) was isolated with the use of a DNA probe generated in a polymerase chain reaction with oligonucleotide primers based on peptide sequences of purified receptor protein. As would be predicted from the cDNA sequence, the LH-CG-R consists of a 26-residue signal peptide, a 341-residue extracellular domain displaying an internal repeat structure characteristic of members of the leucine-rich glycoprotein (LRG) family, and a 333-residue region containing seven transmembrane segments. This membrane-spanning region displays sequence similarity with all members of the G protein-coupled receptor family. Hence, the LH-CG-R gene may have evolved by recombination of LRG and G protein-coupled receptor genes. Cells engineered to express LH-CG-R cDNA bind human choriogonadotropin with high affinity and show an increase in cyclic adenosine monophosphate when exposed to hormone. As revealed by RNA blot analysis and in situ hybridization, the 4.4-kilobase cognate messenger RNA is prominently localized in the rat ovary.


[Post-renal transplant pregnancy: a project to plan carefully].
Kidney transplant is the best treatment for end-stage renal disease (ESRD) as it improves the quality of life and reduces the mortality risk for most patients compared with maintenance dialysis. Additionally, evidence from the literature suggests that renal function, endocrine status and libido rapidly improve after kidney transplant, and one in 50 women of childbearing age become pregnant. Therefore, it seems clear that pregnancy after transplant is a great challenge for physicians involved in this field. The available information on pregnancy outcomes is largely derived from case reports and single-center series, which are unlikely to be representative. Moreover, poor results are less likely to be reported. Many of the reports on long-term outcome show the results of past medical, obstetric, and neonatal care, which may be very different from current practice. Attempts are being made to provide more up-to-date, representative data through national transplantation pregnancy registries. A great number of researchers worldwide have analyzed the biological and endocrinological machinery associated with this event. Additionally, several strategies have been introduced to avoid unplanned pregnancies and to minimize maternal and fetal complications in renal transplant recipients. It seems evident that the return to fertility soon after transplant is often associated with unplanned pregnancy, which can expose both mother and fetus to considerable risks. This underpins the necessity to recommend contraceptive counseling and start clinical follow-up in order to early identify possible pregnancy-related risk factors. In general, pregnancy should not be recommended within the first year after kidney transplant because the risk of acute rejection is greatest and immunosuppressive therapy the most aggressive. It should be planned when organ function and immunosuppressive therapy are stabilized and there is no sign of rejection, hypertension, or chronic infection. Additionally, renal transplant patients and their physicians together must try to identify the best timing, carry out pre-pregnancy screening, and delineate clinical follow-up and future pharmacological programs to minimize or avoid serious maternal and fetal complications. Finally, additional studies are needed to better understand the physiology associated with this condition, improve the pharmacological approach, and analyze the complex ethical and social implications of this important aspect of renal transplantation.


Reward Pays the Cost of Noise Reduction in Motor and Cognitive Control.
Speed-accuracy trade-off is an intensively studied law governing almost all behavioral tasks across species. Here we show that motivation by reward breaks this law, by simultaneously invigorating movement and improving response precision. We devised a model to explain this paradoxical effect of reward by considering a new factor: the cost of control. Exerting control to improve response precision might itself come at a cost--a cost to attenuate a proportion of intrinsic neural noise. Applying a noise-reduction cost to optimal motor control predicted that reward can increase both velocity and accuracy. Similarly, application to decision-making predicted that reward reduces reaction times and errors in cognitive control. We used a novel saccadic distraction task to quantify the speed and accuracy of both movements and decisions under varying reward. Both faster speeds and smaller errors were observed with higher incentives, with the results best fitted by a model including a precision cost. Recent theories consider dopamine to be a key neuromodulator in mediating motivational effects of reward. We therefore examined how Parkinson's disease (PD), a condition associated with dopamine depletion, alters the effects of reward. Individuals with PD showed reduced reward sensitivity in their speed and accuracy, consistent in our model with higher noise-control costs. Including a cost of control over noise explains how reward may allow apparent performance limits to be surpassed. On this view, the pattern of reduced reward sensitivity in PD patients can specifically be accounted for by a higher cost for controlling noise.


The role of footwear in the prevention of foot lesions in patients with NIDDM. Conventional wisdom or evidence-based practice?
To conduct a prospective evaluation of footwear characteristics as predictors of diabetic foot wounds. A total of 352 patients with NIDDM enrolled in a randomized controlled trial aimed at preventing diabetic foot lesions in an academic general medicine practice were studied. Foot wounds (n = 63) were modeled univariately and multivariably using generalized estimating equations. The dependent variable was a wound classified as a 1.2 or greater according to the Seattle Wound Classification System, indicating at least a superficial or healing minor lesion with no functional interruption of the protective cutaneous barrier. Independent variables included detailed measures of style and material of patients' indoor and outdoor shoes, appropriate length and width, sock fibers, whether the patient had bought new shoes in the past 6 months, and if the patient had been recommended for special shoes. Modeling controlled for intervention status and physiological measures (baseline wound, monofilament abnormalities, and serum HDL level). Initial screening (P < 0.20) suggested that a recommendation for special shoes, shoe length, and shoe width were indicative of wounds at follow-up (odds ratios [ORs] 2.19, 1.84, 1.86, respectively), while having bought shoes in the past 6 months was associated with no wound at follow-up (OR 0.60). The final multivariable model included only the recommendation for special shoes (OR 2.19; 95% CI 1.07-4.49). Many variables commonly cited as protective measures in footwear for diabetic patients were not prospectively predictive when controlling for physiological risk factors. Rigorous analyses are needed to examine the many assumptions regarding footwear recommendations for diabetic patients.


Vitamin B complex supplementation as a homocysteine-lowering therapy for early stage diabetic nephropathy in pediatric patients with type 1 diabetes: A randomized controlled trial.
Homocysteine levels are elevated in patients with type 1 diabetes mellitus (T1DM) and could induce renal injury. B vitamins have an important role in preventing microvascular complications of diabetes. We performed a randomized-controlled trial of oral supplementation with vitamin B complex as an adjuvant therapy for nephropathy in pediatric T1DM patients and assessed its relation to homocysteine and cystatin C as a marker of nephropathy. This trial included 80 T1DM patients with microalbuminuria, despite oral angiotensin-converting enzyme inhibitors, aged 12-18 years with at least 5 years disease duration and HbA1c ≤8.5%. Patients were randomly assigned into two groups; intervention group which received oral vitamin B complex (B1, B6 and B12) once daily and placebo group. Both groups were followed-up for 12 weeks with assessment of plasma homocysteine, HbA1c, urinary albumin excretion (UAE) and cystatin C. Both groups were well-matched in baseline clinical and laboratory parameters. Baseline homocysteine levels were elevated in both groups compared with reference control values. After 12 weeks, supplementation with vitamin B complex for the intervention group resulted in a significant decrease of homocysteine, fasting blood glucose, HbA1c, triglycerides, total cholesterol, UAE and cystatin C compared with baseline levels (p < 0.001) and with placebo group (p < 0.001). No adverse reactions were reported. Baseline cystatin C was negatively correlated to vitamin B12 (r = -0.77, p = 0.001). Vitamin B complex improved glycemic control and renal function through decreasing homocysteine and could be a safe and effective strategy for treatment of early stage nephropathy in pediatric T1DM. This trial was registered at ClinicalTrials.gov (NCT03594240).


Mediterranean diet, inflammatory and metabolic biomarkers, and risk of Alzheimer's disease.
We aimed to investigate the association between adherence to the Mediterranean diet (MeDi) and Alzheimer's disease (AD) risk in a prospective study. Specifically, we analyzed reduced inflammation and improved metabolic profile as a potential medium through which the MeDi reduced the risk of AD. During a 4-year follow-up, 118 incident AD cases were identified among the 1219 non-demented elderly (age ≥ 65) subjects who provided dietary information and blood samples at baseline. We used high-sensitivity C-reactive protein (hsCRP) as an index of systemic inflammation, and fasting insulin and adiponectin as indexes of metabolic profile. We investigated whether there was a change in the association between MeDi and incident AD risk when the biomarkers were introduced into multivariable adjusted COX models. Better adherence to MeDi was associated with lower level of hsCRP (p =0.003), but not fasting insulin or adiponectin. Better adherence to MeDi was significantly associated with lower risk for AD: compared to those in the lowest tertile of MeDi, subjects in the highest tertile had a 34% less risk of developing AD (p-for-trend =0.04). Introduction of the hsCRP, fasting insulin, adiponectin, or combinations of them into the COX model did not change the magnitude of the association between MeDi and incident AD. Ultimately, the favorable association between better adherence to MeDi and lower risk of AD did not seem to be mediated by hsCRP, fasting insulin, or adiponectin. Other aspects of inflammatory and metabolic pathways not captured by these biomarkers, or non-inflammatory or non-metabolic pathways, may be relevant to the MeDi-AD association.


Costimulation of CD28- T lymphocytes by 4-1BB ligand.
The interaction of the T cell surface protein CD28 with its ligand, B7-1 or B7-2, provides a critical costimulatory signal for T cell activation. T cells from CD28- mice are deficient in a variety of responses, including those to lectins and allogeneic spleen cells. However, some immune responses do occur in CD28- mice, suggesting the existence of alternate costimulatory pathways. In this work, we show that T cells purified from CD28- mice respond to B lymphomas expressing 4-1BB ligand (4-1BBL), a member of the TNF gene family. This response is inhibited by a soluble form of 4-1BB, the T cell surface receptor for 4-1BBL. Thus, 4-1BBL/4-1BB interaction provides costimulatory signals to T cells independent of signaling through the CD28 receptor. We find that 4-1BBL is inducible on splenic B cells by CD40 ligand/CD40 interaction or by culturing of splenic dendritic cells, treatments that also induce B7 family molecules. CD28- T cells fail to respond in an MLR to resting allogeneic spleen cells. However, treatment of spleen cells with CD40 ligand renders them competent in activation of CD28- T cells. In contrast to results using B lymphomas as APC, soluble 4-1BB fails to inhibit the T cell response to activated spleen cells. This failure of soluble 4-1BB to block an MLR between CD28+ or CD28- T cells and allogeneic spleen cells is in contrast to a previous report with CD28+ cells.


Coalescence between Au nanoparticles as induced by nanocurvature effect and electron beam athermal activation effect.
The coalescence of two single-crystalline Au nanoparticles on surface of amorphous SiOx nanowire, as induced by electron beam irradiation, was in situ studied at room temperature in a transmission electron microscope. It was observed that along with shrinkage of the SiOx nanowire during irradiation, adjacent Au nanoparticles moved around and migrated close to each other. Once the two nanoparticles contacted with each other, a fast, massive atom transportation took place along their contact surface, where a neck region was created. With a further irradiation, the two nanoparticles rotated, aligning their crystal orientations, and gradually coalesced into a larger single-crystalline nanoparticle. The above coalescence process demonstrated an intriguing surface nanowetting ability and nanograin boundary dislocation climb and slip of Au NPs at room temperature as driven by the non-uniformly distributed nanocurvature over the surface of the two contacting nanoparticles as well as the beam-induced instability and soft mode of atomic vibration, which have been underestimated or neglected in the existing theoretical descriptions or simulations.


Chronic exertional compartment syndrome: muscle changes with isometric exercise.
Chronic exertional compartment syndrome (CECS) is a well-documented cause of lower leg pain in active individuals. The pathophysiology is unclear, although it is generally believed to be associated with increased intramuscular pressure, but there is very little information about muscle function in relation to the onset of pain. To investigate strength, fatigue, and recovery of the anterior tibial muscles in CECS patients and healthy subjects during an isometric exercise protocol. Twenty patients and 22 control subjects (mean age 27.6 yr and 33.0 yr, respectively) performed a 20-min isometric exercise protocol consisting of intermittent maximal voluntary contractions (MVC). Central fatigue was evaluated by comparing changes in electrically stimulated (2 s at 50 Hz) and voluntary contraction force before and during the exercise, and then throughout 10 min of recovery. Muscle size was measured by ultrasonography. Pain and cardiovascular parameters were also examined. The absolute MVC forces were similar, but MVC:body mass of the patients was lower (P < 0.05) as was the ratio of MVC to muscle cross-sectional area (P < 0.01). The extent of central and peripheral fatigue was similar in the two groups. The patients reported significantly higher levels of pain during exercise (P < 0.05 at 4 min) and after the first minute of recovery (P < 0.001). An 8% increase in muscle size after exercise was observed for both groups. There were no differences in the cardiovascular responses of the two groups. CECS patients were somewhat weaker than normal but fatigued at a similar rate during isometric exercise. Patients reported higher pain than controls despite comparable changes in muscle size, suggesting that abnormally tight fascia are not the main cause of CECS symptoms.


Glia fuel neurons with locally synthesized ketone bodies to sustain memory under starvation.
During starvation, mammalian brains can adapt their metabolism, switching from glucose to alternative peripheral fuel sources. In the Drosophila starved brain, memory formation is subject to adaptative plasticity, but whether this adaptive plasticity relies on metabolic adaptation remains unclear. Here we show that during starvation, neurons of the fly olfactory memory centre import and use ketone bodies (KBs) as an energy substrate to sustain aversive memory formation. We identify local providers within the brain, the cortex glia, that use their own lipid store to synthesize KBs before exporting them to neurons via monocarboxylate transporters. Finally, we show that the master energy sensor AMP-activated protein kinase regulates both lipid mobilization and KB export in cortex glia. Our data provide a general schema of the metabolic interactions within the brain to support memory when glucose is scarce.


Role of pressure natriuresis in long-term control of renal electrolyte excretion.
If pressure natriuresis is to play an important role in arterial pressure control, renal perfusion pressure must have a long-term effect on urinary sodium excretion. The aim of this study was to quantitate the importance of renal perfusion pressure per se in controlling renal hemodynamics and electrolyte excretion chronically. Female mongrel dogs (n = 6) were instrumented with bilateral renal artery catheters for measurement of renal perfusion pressure and occluders on both renal arteries for servo-control of renal perfusion pressure at different levels; the urinary bladder was split for determination of renal clearances and electrolyte excretion from each kidney separately. Because both kidneys were exposed to the same neurohumoral influences, any changes in renal function could be attributed to differences in renal perfusion pressure between the two kidneys. After 5 days of control, renal perfusion pressure to one kidney was reduced from 86.7 +/- 0.2 to 74.2 +/- 0.6 mm Hg for 12 days, and pressure in the contralateral kidney increased to 91.5 +/- 0.4 mm Hg. Sodium excretion decreased from 41 +/- 2 to 25 +/- 1 mmol/d in the servo-controlled kidney and increased from 41 +/- 1 to 55 +/- 1 mmol/d in the contralateral kidney during 12 days of servo-control. Urine volume, chloride excretion, and potassium excretion exhibited similar patterns during servo-control. In addition, autoregulation of effective renal plasma flow and glomerular filtration rate was relatively well maintained; however, in the low-pressure kidney, glomerular filtration rate was slightly but significantly lower (approximately 8%) than in the contralateral kidney.(ABSTRACT TRUNCATED AT 250 WORDS)


Mutual Effects of Hydrogen Bonding and Polymer Hydrophobicity on Ibuprofen Crystal Inhibition in Solid Dispersions with Poly(N-vinyl pyrrolidone) and Poly(2-oxazolines).
Poly(N-vinyl pyrrolidone) (PVP), poly(2-methyl-2-oxazoline) (PMOZ), poly(2-ethyl-2-oxazoline) (PEOZ), poly(2-n-propyl-2-oxazoline) (PnPOZ), and poly(2-isopropyl-2-oxazoline) (PiPOZ) were used to prepare solid dispersions with ibuprofen (IB), a model poorly-water soluble drug. Dispersions, prepared by solvent evaporation, were investigated using powder X-ray diffractometry, differential scanning calorimetry, and FTIR spectroscopy; hydrogen bonds formed between IB and all polymers in solid dispersions. PMOZ, the most hydrophilic polymer, showed the poorest ability to reduce or inhibit the crystallinity of IB. In contrast, the more hydrophobic polymers PVP, PEOZ, PnPOZ, and PiPOZ provided greater but similar abilities to reduce IB crystallinity, despite the differing polymer hydrophobicity and that PiPOZ is semi-crystalline. These results indicate that crystallinity disruption is predominantly due to hydrogen bonding between the drug molecules and the polymer. However, carrier properties affected drug dissolution, where PnPOZ exhibited lower critical solution temperature that inhibited the release of IB, whereas drug release from other systems was consistent with the degree of ibuprofen crystallinity within the dispersions.


The distribution of mineral density in the cervical vertebral endplates.
Subsidence of various constructs into the vertebral body is a well-known complication in anterior fusion. Information on bone structure is needed, as a basis for improving these procedures. There are, however, no data available on the distribution of mineral density within vertebral endplates. In this study the regional distribution of mineralization within the cervical endplates with respect to endplate orientation (inferior and superior endplate) and level distribution (C3-C7) was examined by means of computed tomographic osteoabsorptiometry (CT-OAM). The distribution of mineralization in 80 cervical endplates of 8 spinal columns (4 male, 4 female, age range 38-62 years) in vertebrae C3-C7 was investigated by CT osteoabsorptiometry (CT-OAM). The subchondral mineralization distribution revealed considerable topographic differences within each endplate, whereby the areas of greatest density were found in the peripheral marginal zones with maxima in the posterolateral surface, whereas mineralization density was much lower in the central areas. The superior endplates showed an additional posteromedial maximum, whereas the inferior endplates showed an additional anterior mineralization maximum. Comparison of the distribution patters of inferior and superior endplates at different levels from C3 to C7 reveals a uniform increase of mineralization in the anterior portions from cranial to caudal. The mineralization distribution showed characteristic reproducible patterns. The maximal values occurred in the posterolateral parts, and can thus be considered a morphological substrate of high long-term loading. This can serve as a basis for improved prosthesis design and the anchorage point for various fusion techniques.


Defects in interstrand cross-link uncoupling do not account for the extreme sensitivity of ERCC1 and XPF cells to cisplatin.
The anticancer drug cisplatin reacts with DNA leading to the formation of interstrand and intrastrand cross-links that are the critical cytotoxic lesions. In contrast to cells bearing mutations in other components of the nucleotide excision repair apparatus (XPB, XPD, XPG and CSB), cells defective for the ERCC1-XPF structure-specific nuclease are highly sensitive to cisplatin. To determine if the extreme sensitivity of XPF and ERCC1 cells to cisplatin results from specific defects in the repair of either intrastrand or interstrand cross-links we measured the elimination of both lesions in a range of nucleotide excision repair Chinese hamster mutant cell lines, including XPF- and ERCC1-defective cells. Compared to the parental, repair-proficient cell line all the mutants tested were defective in the elimination of both classes of adduct despite their very different levels of increased sensitivity. Consequently, there is no clear relationship between initial incisions at interstrand cross-links or removal of intrastrand adducts and cellular sensitivity. These results demonstrate that the high cisplatin sensitivity of ERCC1 and XPF cells likely results from a defect other than in excision repair. In contrast to other conventional DNA cross-linking agents, we found that the repair of cisplatin adducts does not involve the formation of DNA double-strand breaks. Surprisingly, XRCC2 and XRCC3 cells are defective in the uncoupling step of cisplatin interstrand cross-link repair, suggesting that homologous recombination might be initiated prior to excision of this type of cross-link.


Plasmid profiling of members of the family Enterobacteriaceae, lactobacilli, and bifidobacteria to study the transmission of bacteria from mother to infant.
Plasmid profiles of isolates of the family Enterobacteriaceae, lactobacilli, and bifidobacteria cultured from vaginal, oral, and rectal swabs collected from women soon after admission to a maternity hospital were compared with those of strains detected in the feces of their infants. Lactobacilli inhabiting the vaginas of the mothers did not appear to colonize the infant digestive tract, but evidence for the transmission of fecal isolates of the Enterobacteriaceae and bifidobacteria from mother to infant was obtained in four out of five cases. Many of the bifidobacteria isolated were plasmid-free but could be distinguished with biochemical profiles.


The Clinical Characteristics and Outcomes of Patients with Moderate-to-Severe Coronavirus Disease 2019 Infection and Diabetes in Daegu, South Korea.
Coronavirus disease 2019 (COVID-19) is a global pandemic that had affected more than eight million people worldwide by June 2020. Given the importance of the presence of diabetes mellitus (DM) for host immunity, we retrospectively evaluated the clinical characteristics and outcomes of moderate-to-severe COVID-19 in patients with diabetes. We conducted a multi-center observational study of 1,082 adult inpatients (aged ≥18 years) who were admitted to one of five university hospitals in Daegu because of the severity of their COVID-19-related disease. The demographic, laboratory, and radiologic findings, and the mortality, prevalence of severe disease, and duration of quarantine were compared between patients with and without DM. In addition, 1:1 propensity score (PS)-matching was conducted with the DM group. Compared with the non-DM group (n=847), patients with DM (n=235) were older, exhibited higher mortality, and required more intensive care. Even after PS-matching, patients with DM exhibited more severe disease, and DM remained a prognostic factor for higher mortality (hazard ratio, 2.40; 95% confidence interval, 1.38 to 4.15). Subgroup analysis revealed that the presence of DM was associated with higher mortality, especially in older people (≥70 years old). Prior use of a dipeptidyl peptidase-4 inhibitor or a renin-angiotensin system inhibitor did not affect mortality or the clinical severity of the disease. DM is a significant risk factor for COVID-19 severity and mortality. Our findings imply that COVID-19 patients with DM, especially if elderly, require special attention and prompt intensive care.


Mediated effects of insomnia, psychological distress and medication adherence in the association of eHealth literacy and cardiac events among Iranian older patients with heart failure: a longitudinal study.
Given the importance of improving health for patients with heart failure, the present study examined the temporal associations between eHealth literacy, insomnia, psychological distress, medication adherence, quality of life and cardiac events among older patients with heart failure. With a longitudinal design older patients with echocardiography verified heart failure (N=468; 50.4% New York Heart Association class II, mean age 69.3±7.3 years; 238 men) in need of cardiac care at seven Iranian university outpatient clinics went through clinical examinations and completed the following questionnaires at baseline: eHealth literacy scale (eHEALS, assessing eHealth literacy); 5-item medication adherence report scale (MARS-5, assessing medication adherence); Minnesota living with heart failure questionnaire (MLHFQ, assessing quality of life); insomnia severity index (ISI, assessing insomnia); and hospital anxiety and depression scale (HADS, assessing psychological distress). All the patients completed the ISI and HADS again 3 months later; and the MARS-5 6 months later. Also, their cardiac events were collected 18 months later. Three mediation models were then conducted. eHealth literacy had direct and indirect effects (through insomnia and psychological distress) on medication adherence and quality of life. Moreover, eHealth literacy had protecting effects on cardiac events (hazard ratio (HR) 0.53; 95% confidence interval (CI) 0.37, 0.65) through the mediators of insomnia (HR 0.19; 95% CI 0.15, 0.26), psychological distress (HR 0.08; 95% CI 0.05, 0.12) and medication adherence (HR 0.05; 95% CI 0.04, 0.08). As eHealth literacy was a protector for patients with heart failure, healthcare providers may plan effective programmes to improve eHealth literacy for the population. Additional benefits of improving eHealth literacy in heart failure may be decreased insomnia and psychological distress, improved quality of life, as well as decreased cardiovascular events.


Covalent Functionalization by Cycloaddition Reactions of Pristine Defect-Free Graphene.
Based on a low-temperature scanning tunneling microscopy study, we present a direct visualization of a cycloaddition reaction performed for some specific fluorinated maleimide molecules deposited on graphene. Up to now, it was widely admitted that such a cycloaddition reaction can not happen without pre-existing defects. However, our study shows that the cycloaddition reaction can be carried out on a defect-free basal graphene plane at room temperature. In the course of covalently grafting the molecules to graphene, the sp2 conjugation of carbon atoms was broken, and local sp3 bonds were created. The grafted molecules perturbed the graphene lattice, generating a standing-wave pattern with an anisotropy which was attributed to a (1,2) cycloaddition, as revealed by T-matrix approximation calculations. DFT calculations showed that while both (1,4) and (1,2) cycloadditions were possible on free-standing graphene, only the (1,2) cycloaddition could be obtained for graphene on SiC(0001). Globally averaging spectroscopic techniques, XPS and ARPES, were used to determine the modification in the elemental composition of the samples induced by the reaction, indicating an opening of an electronic gap in graphene.


Isotypic analysis, antigen specificity, and inhibitory function of maternally transmitted Plasmodium falciparum-specific antibodies in Gabonese newborns.
An analysis of Plasmodium falciparum-specific antibodies was performed in pairs of maternal and cord sera from Gabon, a region endemic for malaria. All paired sera (n = 59) had P. falciparum-specific antibodies. Immunofluorescence assays detected parasite-specific IgG1, IgG2, and IgG3 in 100% of the tested pairs (n = 26) and IgG4 in 42% of them. The titers of specific IgG2 and IgG3 were significantly lower in cord than in maternal sera. All maternal sera had specific IgM. Of the seven P. falciparum-IgM positive cord sera, six were associated with malaria-related histological placental changes (MRHPC). In addition, higher titers of specific IgG1 in maternal and cord sera and of specific IgG3 in cord sera were associated with MRHPC. Similar P. falciparum antigens were recognized by cord and corresponding maternal sera in radioimmunoprecipitation and Western blot assays (n = 40). Sixteen of 20 cord sera and 15 of 20 paired maternal sera significantly inhibited in vitro parasite growth. The extent of inhibition did not correlate with the titer of specific antibodies. These data confirm the very effective placental transfer of anti-malarial antibodies. The presence of IgM in some cord sera raise the question of intrauterine sensitization to malaria antigens.


Optimization of operational variables and kinetic modeling for photocatalytic removal of Direct Blue 14 from aqueous media by ZnS nanoparticles.
Zinc sulfide nanoparticles (ZnS-NPs) were synthesized via a simple and facile co-precipitation method and were characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM) and diffuse reflectance spectroscopy (DRS). Photocatalytic activity of synthesized nanoparticles was evaluated in removal of double azo dye Direct Blue 14 (DB14) from aqueous media. Optimization of photocatalytic removal of DB14 was studied using response surface methodology (RSM). Based on the results, DB14 removal efficiency increased with increasing intensity and duration of UV light irradiation, whereas the higher pH and higher initial dye concentration were unfavorable. Under optimum conditions (initial DB14 concentration =10 mg L-1, ZnS-NPs amount = 0.7 g L-1, pH = 3.5, UV light intensity =16 W m-2, and irradiation time = 48 min), dye removal efficiency reached up to 88.26%. In continuation of our researches, non-linear regression analysis was used to development a kinetics model based on the Langmuir-Hinshelwood model and an empirical equation was obtained for estimation of apparent pseudo-first-order rate constant (kap) as a function of the operational variables. Findings indicated a high similarity was between the model prediction and experimental results.


Accidental hypoglycaemia caused by an arterial flush drug error: a case report and contributory causes analysis.
In 2008, the National Patient Safety Agency (NPSA) issued a Rapid Response Report concerning problems with infusions and sampling from arterial lines. The risk of blood sample contamination from glucose-containing arterial line infusions was highlighted and changes in arterial line management were recommended. Despite this guidance, errors with arterial line infusions remain common. We report a case of severe hypoglycaemia and neuroglycopenia caused by glucose contamination of arterial line blood samples. This case occurred despite the implementation of the practice changes recommended in the 2008 NPSA alert. We report an analysis of the factors contributing to this incident using the Yorkshire Contributory Factors Framework. We discuss the nature of the errors that occurred and list the consequent changes in practice implemented on our unit to prevent recurrence of this incident, which go well beyond those recommended by the NPSA in 2008.


Typing Candida Species Using Microsatellite Length Polymorphism and Multilocus Sequence Typing.
To gain more insight into the epidemiological relationships between isolates of Candida spp. obtained from various origins, several molecular typing techniques have been developed. Two methods have emerged in the 2000s as soon as enough knowledge of the Candida spp. genomes was available to choose adequate loci and primers, namely microsatellite length polymorphism (MLP) and multilocus sequence typing (MLST). To contrast with previous PCR-based methods, specific amplifications with stringent conditions easily reproducible are the basis of MLP and MLST. MLST relies on Sanger sequencing to detect single-nucleotide polymorphisms within housekeeping genes. MLP needs a first in silico step to select tandemly repeated stretches of two to five nucleotides. One of the two primers used to amplify a microsatellite locus is labeled and fragment sizing is automatically performed using high-resolution electrophoresis platforms. MLST provides results easily comparable between laboratories and active MLST schemes are publicly available for the main Candida species. For comparative studies, MLP needs standards to compensate for the electrophoretic variations depending on the platforms used. Both methods can help us gain insight into the genetic relatedness of fungal isolates, both with advantages and drawbacks, and the choice of one method rather than the other depends on the task in question.


A pilot study of the efficacy of varenicline for the treatment of smokeless tobacco users in Midwestern United States.
Long-term smokeless tobacco (ST) use is known to increase the risk for oropharyngeal cancer, heart attack, and stroke. Varenicline has recently been demonstrated to increase ST abstinence rates among Swedish snus users. We have conducted a pilot study to obtain preliminary evidence of efficacy of varenicline for the treatment of ST users in Midwestern United States. We conducted a randomized, placebo-controlled Phase II clinical trial to evaluate the potential efficacy of 12 weeks of varenicline for the treatment of ST users with an a priori decision rule that a 1-tailed p < .20 for the comparison of the primary endpoint was evidence to conclude that future studies were warranted. Subjects were followed for 6 months after randomization. We randomized 76 subjects (38 varenicline and 38 placebo). Subjects were similar at baseline with a mean age of 41 years, and all were male. The biochemically confirmed point prevalence tobacco abstinence rates at end of treatment were 55.3% for varenicline and 42.1% for placebo (p = .126) and 47.4% and 31.6% (p = .080), respectively, at 6 months. Point prevalence ST abstinence rates at end of treatment for varenicline were 57.9% and 42.1% for placebo (p = .084) and 57.9% and 31.6% (p = .011), respectively, at 6 months. Varenicline was associated with significantly less craving compared with placebo. Varenicline was well tolerated with nausea and sleep disturbance being the most common side effects. Varenicline decreases craving and may be effective for increasing tobacco abstinence rates among ST users. Larger trials may be warranted to confirm these results.


Induction of rules for biological macromolecule crystallization.
X-ray crystallography is the method of choice for determining the 3-D structure of large macromolecules at a high enough resolution. The rate limiting step in structure determination is the crystallization itself. It takes anywhere between a few weeks to several years to obtain macromolecular crystals that yield good diffraction patterns. The theory of forces that promote and maintain crystal growth is preliminary, and crystallographers systematically search a large parameter space of experimental settings to grow good crystals. There is a wealth of experimental data on crystal growth most of which is in paper laboratory notebooks. Some of the data has been gathered in electronic form, e.g., the Biological Macromolecular Crystallization Database (BMCD) which is a repository of successful experimental conditions for growing over 800 different macromolecules (Gilliland 1987). Crystallographers are in need of computational tools to gather and analyze past data to design new crystal growth trails. We are building the Crystallographer's Assistant (CA) to help crystallographers record and maintain experimental context in electronic form, offer suggestions on experimental conditions that are likely to be successful, and provide explanations for failed experiments. As an initial step in this project, we have applied RL, an inductive learning program, to the BMCD. In this paper we report initial experiments and findings in applying RL to the BMCD. From the point of view of crystallography, we have discovered possibly significant new empirical relationships in crystal growth. From the point of view of machine learning, our work suggests refinements of existing methods for incorporating detailed domain knowledge into inductive analysis techniques.


Investigational luteinizing hormone releasing hormone (LHRH) agonists and other hormonal agents in early stage clinical trials for prostate cancer.
The treatment and management of prostate cancer continues to evolve; newer classes of agents and combination therapies are being developed and some are being investigated in early phase clinical trials. We discuss investigational hormonal agents for the treatment of prostate cancer and focus primarily on luteinizing hormone releasing hormone (LHRH) agonists in early stage trials. We look at agents that target the hormonal axis, including anti-androgens, gonadotropins, estrogenic agents and progestogenic agents and other non-hormonal agents often used in combination with LHRH agonists. We review these candidates in the specific clinical niche in which they might find utility. Of all candidate compounds being evaluated in clinical trials, very few will receive FDA approval. Few, if any of the investigational agents discussed here will be used routinely in clinical practice for treating prostate cancer. Recognizing the reasons for the failure of agents to advance to later stage trials is important. Furthermore, a thorough understanding of the mechanisms underlying prostate cancer pathogenesis, including various points in the HGPA and parallel pathways, will help identify potentially actionable targets.


A system of two polymerases--a model for the origin of life.
What was the first living molecule--RNA or protein? This question embodies the major disagreement in studies on the origin of life. The fact that in contemporary cells RNA polymerase is a protein and peptidyl transferase consists of RNA suggests the existence of a mutual catalytic dependence between these two kinds of biopolymers. I suggest that this dependence is a 'frozen accident', a remnant from the first living system. This system is proposed to be a combination of an RNA molecule capable of catalyzing amino acid polymerization and the resulting protein functioning as an RNA-dependent RNA polymerase. The specificity of the protein synthesis is thought to be achieved by the composition of the surrounding medium and the specificity of the RNA synthesis--by Watson-Crick base pairing. Despite its apparent simplicity, the system possesses a great potential to evolve into a primitive ribosome and further to life, as it is seen today. This model provides a possible explanation for the origin of the interaction between nucleic acids and protein. Based on the suggested system, I propose a new definition of life as a system of nucleic acid and protein polymerases with a constant supply of monomers, energy and protection.


Re-conceptualizing the origins of life.
Over the last several hundred years of scientific progress, we have arrived at a deep understanding of the non-living world. We have not yet achieved an analogous, deep understanding of the living world. The origins of life is our best chance at discovering scientific laws governing life, because it marks the point of departure from the predictable physical and chemical world to the novel, history-dependent living world. This theme issue aims to explore ways to build a deeper understanding of the nature of biology, by modelling the origins of life on a sufficiently abstract level, starting from prebiotic conditions on Earth and possibly on other planets and bridging quantitative frameworks approaching universal aspects of life. The aim of the editors is to stimulate new directions for solving the origins of life. The present introduction represents the point of view of the editors on some of the most promising future directions.This article is part of the themed issue 'Reconceptualizing the origins of life'.


Neoclassical tearing physics in the spherical tokamak MAST.
Results from MAST provide a first test of neoclassical tearing mode physics in the spherical tokamak (ST). The mode accounts for the main performance limit in conventional tokamaks. Its behavior in the ST is remarkably well described by existing theoretical models, although it is more readily seeded by sawtooth events in these scenarios. Modeling confirms the significance of stabilizing field-curvature effects. This provides good grounds for optimism that with suitable control of profiles, it may be possible to avoid these modes in the ST.


A decrease in brain activation associated with driving when listening to someone speak.
Behavioral studies have shown that engaging in a secondary task, such as talking on a cellular telephone, disrupts driving performance. This study used functional magnetic resonance imaging (fMRI) to investigate the impact of concurrent auditory language comprehension on the brain activity associated with a simulated driving task. Participants steered a vehicle along a curving virtual road, either undisturbed or while listening to spoken sentences that they judged as true or false. The dual-task condition produced a significant deterioration in driving accuracy caused by the processing of the auditory sentences. At the same time, the parietal lobe activation associated with spatial processing in the undisturbed driving task decreased by 37% when participants concurrently listened to sentences. The findings show that language comprehension performed concurrently with driving draws mental resources away from the driving and produces deterioration in driving performance, even when it does not require holding or dialing a phone.


The effectiveness of video-telemedicine for screening of patients requesting emergency air medical transport (EAMT).
Demand for emergency air medical transport (EAMT) services have increased in recent years. However, the high costs of these services have raised questions on the benefit to patient outcomes. In this study, we evaluate the effectiveness of video-telemedicine for the preflight screening of patients for air medical transports. A prospective cohort study. Medical records of patients transported from the Penghu Islands to Taiwan were retrospectively collected from November 1999 to October 2002 (stage 1). In addition, we collected medical records of patients who were preflight-screened by physicians using video Web cameras from November 1, 2002 through August 30, 2003 (stage 2). The intervention in stage 2 included a set of protocols and screening criteria for EAMT implemented by the National Aeromedical Consultation Center (NACC). In stage 1, there were no standardized protocols or screening guidelines for EAMT. The EAMT system before implementing preflight screening and telemedicine was mostly based on patient's requests and their health condition determined by the treating medical officers (TMO). A total of 822 transfers were included in this study. Patient demographic backgrounds in the two groups were similar on gender, age, disease classification, and types of illnesses. Patients in stage 2 were significantly older than those in stage 1. In a comparison of flight frequencies between the two stages, the results revealed a 36.2% reduction of EAMT applications in stage 2. The flight approval rate was 91.2%. The intervention in stage 2 also presented a significant reduction in cross-zone transport (16.1% to 0.1% to the northern Taiwan region). Within-zone transfers increased from 74.9% to 88.3%. Cost analysis showed that physician triage in stage 2 resulted in a total annual savings on EAMTs of US 448,986 dollars. This study demonstrates the physician-assisted preflight screening using video-telemedicine significantly reduced the frequency of unnecessary air medical transports and consequently led to reduced costs. Video-telemedicine can be an essential tool to support physicians in decision-making for patient screening.


Renal protective effect of Paeoniflorin by inhibition of JAK2/STAT3 signaling pathway in diabetic mice.
Paeoniflorin is the main bioactive components of the root of P.lactiflora Pall., and has been widely used as an anti-inflammation and immunomodulatory agent. However, the effect and mechanisms of Paeoniflorin in diabetic nephropathy (DN) remains to be elucidated. In the present study, streptozotocin (STZ)-induced type 1 diabetic mice model was used to investigate the protective effect of Paeoniflorin and the role of the Janus kinase (JAK) 2/signal transducer (STAT) 3 signaling pathway on DN. After treatment with Paeoniflorin at a dose of 25, 50 and 100 mg/kg once a day for 12 weeks, both the functional and histological damage to diabetic mice kidney had been attenuated significantly. Additionally, these reno-protective effects were associated with alleviating macrophage infiltration and inflammatory factors expression as well as suppression of the JAK2/STAT3 signaling pathway. These data reveal that Paeoniflorin attenuates renal lesions in diabetic mice and these protective effects may be associated with the prevention of macrophage infiltration and inhibition of the JAK2/STAT3 signaling pathway.


Craniopharyngiomas in children and adults: systematic analysis of 121 cases with long-term follow-up.
Craniopharyngiomas account for 2-5% of all primary intracranial tumours. Despite their benign histological appearance, they are often associated with an unfavourable prognosis and their optimal treatment remains controversial. To analyse the natural history and treatment outcome of children and adults presenting to the Departments of Paediatrics and Endocrinology with craniopharyngioma between 1964 and 2003. The records of 121 patients (age range 2.5-83 years, 42 aged < 16 and 79 aged > or = 16) were identified. The mean follow-up period since presentation was 103 months (8.6 years) (range 0.3-468 months). Sixteen patients underwent gross total removal (A), 3 gross total removal + radiotherapy (B), 51 partial removal (C), 33 partial removal + radiotherapy (D), 6 cyst evacuation alone (E) and 3 cyst evacuation + radiotherapy (F). The clinical, imaging and endocrinological data at presentation and during follow-up were analysed. Headache and visual field defects were the most common presenting clinical features (64% and 55%, respectively). Ninety-four per cent of the tumours had an extrasellar component and 23% of them were associated with hydrocephalus. There was a significant difference in the recurrence-free survival rates between groups A-D [at 10 years: 100% (A), 100% (B), 38% (C) and 77% (D), P < 0.01], which persisted even when analysing patients operated after 1980. The median time of first recurrence was 2.5 years (range 0.5-36). The peri-operative mortality of the patients who had any type of neurosurgical intervention due to recurrence was higher than that observed after primary surgery (24%vs. 1.8%) (P < 0.01). The rate of re-accumulation of the cyst fluid was 58% during the first year in patients of group E, whereas none of the subjects of group F experienced such an event during their follow-up period. There was no reversal of pre-existing pituitary hormone deficits after any surgical intervention. The probabilities of GH, FSH/LH, ACTH, TSH deficiency and diabetes insipidus at the 10-year follow-up were 88%, 90%, 86%, 80% and 65%, respectively. After excluding the non-tumour-related deaths, the 10-year survival rate following presentation was 90%. Patients with recurrence had a significantly lower probability for survival compared with those without it (at 10 years: 70%vs. 99%, P < 0.01). At the 10-year follow-up the probability of the presence of major visual field defects was 48%, hyperphagia/obesity 39%, epilepsy 12% and hemi-/monoparesis 11%. In this large series no substantial differences in the outcome of tumours diagnosed during childhood or adult life were found. Craniopharyngiomas remain tumours associated with significant morbidity. Gross total removal provides favourable results in terms of recurrences. If this cannot be achieved safely, adjuvant radiotherapy is beneficial in preventing tumour re-growth. Childhood- and adult-onset lesions generally behave similarly.


A bacterial inhibitor of host programmed cell death defenses is an E3 ubiquitin ligase.
The Pseudomonas syringae protein AvrPtoB is translocated into plant cells, where it inhibits immunity-associated programmed cell death (PCD). The structure of a C-terminal domain of AvrPtoB that is essential for anti-PCD activity reveals an unexpected homology to the U-box and RING-finger components of eukaryotic E3 ubiquitin ligases, and we show that AvrPtoB has ubiquitin ligase activity. Mutation of conserved residues involved in the binding of E2 ubiquitin-conjugating enzymes abolishes this activity in vitro, as well as anti-PCD activity in tomato leaves, which dramatically decreases virulence. These results show that Pseudomonas syringae uses a mimic of host E3 ubiquitin ligases to inactivate plant defenses.


Natural variation in a glucuronosyltransferase modulates propionate sensitivity in a C. elegans propionic acidemia model.
Mutations in human metabolic genes can lead to rare diseases known as inborn errors of human metabolism. For instance, patients with loss-of-function mutations in either subunit of propionyl-CoA carboxylase suffer from propionic acidemia because they cannot catabolize propionate, leading to its harmful accumulation. Both the penetrance and expressivity of metabolic disorders can be modulated by genetic background. However, modifiers of these diseases are difficult to identify because of the lack of statistical power for rare diseases in human genetics. Here, we use a model of propionic acidemia in the nematode Caenorhabditis elegans to identify genetic modifiers of propionate sensitivity. Using genome-wide association (GWA) mapping across wild strains, we identify several genomic regions correlated with reduced propionate sensitivity. We find that natural variation in the putative glucuronosyltransferase GLCT-3, a homolog of human B3GAT, partly explains differences in propionate sensitivity in one of these genomic intervals. We demonstrate that loss-of-function alleles in glct-3 render the animals less sensitive to propionate. Additionally, we find that C. elegans has an expansion of the glct gene family, suggesting that the number of members of this family could influence sensitivity to excess propionate. Our findings demonstrate that natural variation in genes that are not directly associated with propionate breakdown can modulate propionate sensitivity. Our study provides a framework for using C. elegans to characterize the contributions of genetic background in models of human inborn errors in metabolism.


Abrogation of ospAB constitutively activates the Rrp2-RpoN-RpoS pathway (sigmaN-sigmaS cascade) in Borrelia burgdorferi.
Molecular mechanisms underlying the reciprocal regulation of the two major surface lipoproteins and virulence factors of Borrelia burgdorferi, OspA and OspC, are not fully understood. Herein, we report that inactivation of the ospAB operon resulted in overproduction of OspC and many other lipoproteins via the constitutive activation of the Rrp2-RpoN-RpoS pathway. Complementing the ospAB mutant with a wild-type copy of ospA, but not an ospA variant that lacks the lipoprotein signal sequence, restored normal regulation of the Rrp2-RpoN-RpoS pathway; these results indicate that the phenotype was not caused by spurious mutations. Interestingly, while most of the ospAB mutant clones displayed a constitutive ospC expression phenotype, some ospAB mutant clones showed little or no ospC expression. Further analyses revealed that this OspC-negative phenotype was independent of abrogation of ospAB. While activation of the Rrp2-RpoN-RpoS pathway was recently shown to downregulate ospA, our findings suggest that reduction of OspA can also activate this pathway. We postulate that the activation of the Rrp2-RpoN-RpoS pathway and downregulation of OspA form a positive feedback loop that allows spirochaetes to produce and maintain a constant high level of OspC and other lipoproteins during tick feeding, a strategy that is critical for spirochaetal transmission and mammalian infection.


Heritability of circle of Willis variations in families with intracranial aneurysms.
Intracranial aneurysms more often occur in the same arterial territory within families. Several aneurysm locations are associated with specific circle of Willis variations. We investigated whether the same circle of Willis variations are more likely to occur in first-degree relatives than in unrelated individuals. We assessed four circle of Willis variations (classical, A1-asymmetry, incomplete posterior communicating artery and fetal circulation) in two independent groups of families with familial aneurysms and ≥2 first-degree relatives with circle of Willis imaging on MRA/CTA. In each (index) family we determined the proportion of first-degree relatives with the same circle of Willis variation as the proband and compared it to the proportion of first-degree relatives of a randomly selected unrelated (comparison) family who had the same circle of Willis variation as the index family's proband. Concordance in index families and comparison families was compared with a conditional logistic events/trials model. The analysis was simulated 1001 times; we report the median concordances, odds ratios (ORs), and 95% confidence intervals (95%CI). The groups were analysed separately and together by meta-analysis. We found a higher overall concordance in circle of Willis configuration in index families than in comparison families (meta-analysis, 244 families: OR 2.2, 95%CI 1.6-3.0) mostly attributable to a higher concordance in incomplete posterior communicating artery (meta-analysis: OR 2.8, 95%CI 1.8-4.3). No association was found for the other three circle of Willis variations. In two independent groups of families with familial aneurysms, the incomplete PcomA variation occurred more often within than between families suggesting heritability of this circle of Willis variation. Further studies should investigate genetic variants associated with circle of Willis formation.


Erlotinib/gemcitabine for first-line treatment of locally advanced or metastatic adenocarcinoma of the pancreas.
Erlotinib (Tarceva) is a human epidermal growth factor receptor type 1/epidermal growth factor receptor (HER1/EGFR) tyrosine kinase inhibitor initially approved by the US Food and Drug Administration for the treatment of patients with locally advanced or metastatic non-small-cell lung cancer after failure of at least one prior chemotherapy regimen. In this report, we present the pivotal study that led to the approval of erlotinib in combination with gemcitabine (Gemzar) in patients with locally advanced/metastatic chemonaive pancreatic cancer patients. The combination demonstrated a statistically significant increase in overall survival accompanied by an increase in toxicity. Physicians and patients now have a new option for the treatment of locally advanced/metastatic adenocarcinoma of the pancreas.


Poly-arginine R18 and R18D (D-enantiomer) peptides reduce infarct volume and improves behavioural outcomes following perinatal hypoxic-ischaemic encephalopathy in the P7 rat.
We examined the neuroprotective efficacy of the poly-arginine peptide R18 and its D-enantiomer R18D in a perinatal hypoxic-ischaemic (HI) model in P7 Sprague-Dawley rats. R18 and R18D peptides were administered intraperitoneally at doses of 30, 100, 300 or 1000 nmol/kg immediately after HI (8% O2/92%N2 for 2.5 h). The previously characterised neuroprotective JNKI-1-TATD peptide at a dose of 1000 nmol/kg was used as a control. Infarct volume and behavioural outcomes were measured 48 h after HI. For the R18 and R18D doses examined, total infarct volume was reduced by 25.93% to 43.80% (P = 0.038 to < 0.001). By comparison, the JNKI-1-TATD reduced lesion volume by 25.27% (P = 0.073). Moreover, R18 and R18D treatment resulted in significant improvements in behavioural outcomes, while with JNKI-1-TATD there was a trend towards improvement. As an insight into the likely mechanism underlying the effects of R18, R18D and JNKI-1-TATD, the peptides were added to cortical neuronal cultures exposed to glutamic acid excitotoxicity, resulting in up to 89, 100 and 71% neuroprotection, respectively, and a dose dependent inhibition of neuronal calcium influx. The study further confirms the neuroprotective properties of poly-arginine peptides, and suggests a potential therapeutic role for R18 and R18D in the treatment of HIE.


Orbital complications of pediatric rhinosinusitis: A single institution report.
Rhinosinusitis is one of the most common infections of childhood. Rhinosinusitis usually limits itself in the pediatric age group, however orbital and intracranial complications may arise in some of the patients. The purpose of the study was to retrospectively analyse the previous treatments and outcomes in pediatric rhinosinusitis patients with orbital complications. The effect and prognosis of medical treatment and endoscopic sinus surgery are reported in this study. Twenty-five pediatric patients diagnosed with complicated rhinosinusitis between January 2008 - February 2016 were included in the study. Orbital complications, examination findings, radiological imaging and blood counts were retrospectively collected from patient files. The duration of hospitalization, source of the infection, complications, previous medical and surgical treatments were also retrospectively assessed. Average age of the patients were 8.84 ± 4.02 years (range: 1-16 years). The mean length of stay in hospital was 6.72 ± 3.28 days. Hospitalization in surgically treated group was higher than primarily medically treated group. However this difference could not reach to a statistically significant level (p > 0.05). Mean hospitalization time was found 5.21 ± 2.51 and 8.43 ± 2.87 days in patients diagnosed with preseptal cellulitis and subperiosteal abscess respectively. Hospitalization in patients with subperiosteal abscess was higher than preseptal cellulitis and a statistically significant difference was detected (p < 0.05). Morbidity and mortality of orbital complications which are the most common complications of pediatric rhinosinusitis, could significantly be reduced by using appropriate treatment methods and an early diagnosis. Conservative therapy is an effective method for patients with preseptal cellulitis and most cases of orbital cellulitis in children.


Pseudomonas Blight Caused by Pseudomonas syringae on Raspberry in California.
In February 2013 in coastal California (Santa Cruz County), plantings of red raspberry (Rubus idaeus var. strigosus) exhibited symptoms of a previously undocumented disease. Initial symptoms were small (less than 5 mm wide), angular, water-soaked lesions on leaf and petiole tissues of recently emerged foliage. Lesions were observable from both adaxial and abaxial leaf surfaces. As disease progressed, lesions enlarged and coalesced, resulting in significant dark brown to black blighting of the foliage. The foliage of severely affected plants was stunted and wilted. The disease affected 5 ha and incidence was approximately 30%. Cream-colored bacterial colonies were isolated from surface disinfested symptomatic tissue that was macerated and streaked onto King's medium B (KMB) and sucrose peptone agar (SPA). Fungi were not recovered from any tissue that was surface disinfested and placed into acidified potato dextrose agar. Four representative strains were fluorescent on KMB and gram-negative based on lysis by KOH. Strains were positive for levan formation, negative for oxidase and arginine dihydrolase, and did not cause soft rot on potato slices but induced a hypersensitive response in tobacco (Nicotiana tabacum L. cv. Samsun); strains thus belonged to Lelliot's LOPAT group 1, P. syringae (3). All four strains had identical DNA fragment-banding patterns generated by repetitive extragenic palindromic sequence (rep)-PCR using the BOXA1R primer (4). The pattern generated was different than all P. syringae pathovars in genomospecies 1 including P. syringae pv. syringae. According to multilocus sequence analysis conducted by previously described methods, the strains are most closely related to P. syringae pv. aceris and P. syringae pv. solidagae in genomospecies 1 (1). Potted raspberry plants were used to test four strains for pathogenicity. Inoculum was prepared by growing the bacteria on SPA for 48 h and suspending the bacteria in sterile distilled water (SDW) for a final concentration of approximately 107 CFU/ml. Suspensions were sprayed until runoff onto three replicate plants per strain. Control plants were sprayed with SDW until runoff. Plants were enclosed in plastic bags for 24 h and then maintained in a greenhouse (23 to 25°C). After 7 to 8 days, water soaked lesions developed on all inoculated plants; lesions later turned dark brown and appeared similar to symptoms observed in the field. Plants treated with water developed no symptoms. Bacteria re-isolated onto KMB from symptomatic tissues were fluorescent and appeared identical to the bacteria used to inoculate the plants; two selected re-isolated strains were identical to the original strains according to rep-PCR, fluorescence, and LOPAT reactions. The experiment was repeated and disease development and recovery of fluorescent strains on KMB was identical to the first experiment. To our knowledge, this is the first report of Pseudomonas blight of raspberry, caused by P. syringae, in California. Affected plants initially were stunted in growth but later in the summer exhibited no lasting effects from the disease. Pseudomonas blight has been reported in the Pacific Northwest region of the United States, the British Columbia region of Canada, and Serbia (2). References: (1) C. T. Bull et al. Phytopathology 101:847, 2011. (2) Z. Ivanovic et al. Eur. J. Plant Pathol. 134:191, 2012. (3) R. A. Lelliott. J. Appl. Bacteriol. 29:470, 1966. (4) A. S. A. Marques, et al. Genet. Mol. Biol. 31:106. 2008.


Identification of Deregulated Pathways, Key Regulators, and Novel miRNA-mRNA Interactions in HPV-Mediated Transformation.
Next to a persistent infection with high-risk human papillomavirus (HPV), molecular changes are required for the development of cervical cancer. To identify which molecular alterations drive carcinogenesis, we performed a comprehensive and longitudinal molecular characterization of HPV-transformed keratinocyte cell lines. Comparative genomic hybridization, mRNA, and miRNA expression analysis of four HPV-containing keratinocyte cell lines at eight different time points was performed. Data was analyzed using unsupervised hierarchical clustering, integrated longitudinal expression analysis, and pathway enrichment analysis. Biological relevance of identified key regulatory genes was evaluated in vitro and dual-luciferase assays were used to confirm predicted miRNA-mRNA interactions. We show that the acquisition of anchorage independence of HPV-containing keratinocyte cell lines is particularly associated with copy number alterations. Approximately one third of differentially expressed mRNAs and miRNAs was directly attributable to copy number alterations. Focal adhesion, TGF-beta signaling, and mTOR signaling pathways were enriched among these genes. PITX2 was identified as key regulator of TGF-beta signaling and inhibited cell growth in vitro, most likely by inducing cell cycle arrest and apoptosis. Predicted miRNA-mRNA interactions miR-221-3p_BRWD3, miR-221-3p_FOS, and miR-138-5p_PLXNB2 were confirmed in vitro. Integrated longitudinal analysis of our HPV-induced carcinogenesis model pinpointed relevant interconnected molecular changes and crucial signaling pathways in HPV-mediated transformation.


Foam-oil interaction in porous media: implications for foam assisted enhanced oil recovery.
The efficiency of a foam displacement process in enhanced oil recovery (EOR) depends largely on the stability of foam films in the presence of oil. Experimental studies have demonstrated the detrimental impact of oil on foam stability. This paper reviews the mechanisms and theories (disjoining pressure, coalescence and drainage, entering and spreading of oil, oil emulsification, pinch-off, etc.) suggested in the literature to explain the impact of oil on foam stability in the bulk and porous media. Moreover, we describe the existing approaches to foam modeling in porous media and the ways these models describe the oil effect on foam propagation in porous media. Further, we present various ideas on an improvement of foam stability and longevity in the presence of oil. The outstanding questions regarding foam-oil interactions and modeling of these interactions are pointed out.


Growth of Lactobacillus paracasei ATCC 334 in a cheese model system: a biochemical approach.
Growth of Lactobacillus paracasei ATCC 334, in a cheese-ripening model system based upon a medium prepared from ripening Cheddar cheese extract (CCE) was evaluated. Lactobacillus paracasei ATCC 334 grows in CCE made from cheese ripened for 2 (2mCCE), 6 (6mCCE), and 8 (8mCCE) mo, to final cell densities of 5.9×10(8), 1.2×10(8), and 2.1×10(7)cfu/mL, respectively. Biochemical analysis and mass balance equations were used to determine substrate consumption patterns and products formed in 2mCCE. The products formed included formate, acetate, and D-lactate. These data allowed us to identify the pathways likely used and to initiate metabolic flux analysis. The production of volatiles during growth of Lb. paracasei ATCC 334 in 8mCCE was monitored to evaluate the metabolic pathways utilized by Lb. paracasei during the later stages of ripening Cheddar cheese. The 2 volatiles detected at high levels were ethanol and acetate. The remaining detected volatiles are present in significantly lower amounts and likely result from amino acid, pyruvate, and acetyl-coenzyme A metabolism. Carbon balance of galactose, lactose, citrate, and phosphoserine/phosphoserine-containing peptides in terms of D-lactate, acetate, and formate are in agreement with the amounts of substrates observed in 2mCCE; however, this was not the case for 6mCCE and 8mCCE, suggesting that additional energy sources are utilized during growth of Lb. paracasei ATCC 334 in these CCE. This study provides valuable information on the biochemistry and physiology of Lb. paracasei ATCC 334 in ripening cheese.


Role of endoplasmic reticulum calcium disequilibria in the mechanism of homocysteine-induced ER stress.
Our laboratory demonstrated that hyperhomocysteinemia accelerates atherosclerosis in mouse models through ER stress and activation of the unfolded protein response (UPR). In this study, we tested the hypothesis that homocysteine-induced ER stress may arise from ER-Ca(2+) disequilibria. We found that homocysteine-induced cytosolic Ca(2+) transients in T24/83 cells and human aortic smooth muscle cells (HASMCs). These calcium effects occurred at concentrations of homocysteine in the external medium (1-5 mM) that increase intracellular homocysteine in these cell types. Prolonged homocysteine treatment (5 h) at these exogenous concentrations reduced ER-Ca(2+) emptying evoked by thapsigargin. However, these homocysteine-induced effects on ER-Ca(2+) emptying were of a much smaller magnitude than those evoked by A23187 or thapsigargin (ER stressors known to induce ER stress through ER-Ca(2+) depletion). T24/83 cells stably overexpressing the Ca(2+)-binding ER chaperone GRP78 showed diminished cytosolic Ca(2+) transients induced by homocysteine and reduced ER-Ca(2+) emptying evoked by thapsigargin. Prevention of the homocysteine-induced UPR by cycloheximide pretreatment normalized GRP78 expression and ER-Ca(2+) emptying evoked by thapsigargin. These results are inconsistent with a mechanism of ER stress induction by homocysteine through ER-Ca(2+) depletion.


Violent media exposure, aggression and CU traits in adolescence: Testing the selection and socialization hypotheses.
We investigated the role of exposure to violent action for later aggression and for later callous-unemotional traits in a sample of Swedish adolescents (N = 77-85), testing the selection and socialization hypotheses. Adolescents reported on violent delinquency and on callous-unemotional (CU) traits at age 15, on their media habits at age 16 and on reactive and proactive aggression and CU traits at age 18. The socialization hypothesis was supported with regard to aggression, that is, violent delinquency did not affect consumption of violent action, but controlling for violent delinquency, consumption of violent action added to proactive aggression and, marginally, to reactive aggression. The selection hypothesis was supported with regard to CU traits, that is, high levels of CU traits predicted frequent consumption of violent action, but consumption of violent action did not affect later levels of CU traits. Frequent violent media use was associated with later aggression. The associations between CU traits and violent media need further study.


Perturbing the ubiquitin pathway reveals how mitosis is hijacked to denucleate and regulate cell proliferation and differentiation in vivo.
The eye lens presents a unique opportunity to explore roles for specific molecules in cell proliferation, differentiation and development because cells remain in place throughout life and, like red blood cells and keratinocytes, they go through the most extreme differentiation, including removal of nuclei and cessation of protein synthesis. Ubiquitination controls many critical cellular processes, most of which require specific lysines on ubiquitin (Ub). Of the 7 lysines (K) least is known about effects of modification of K6. We replaced K6 with tryptophan (W) because K6 is the most readily modified K and W is the most structurally similar residue to biotin. The backbone of K6W-Ub is indistinguishable from that of Wt-Ub. K6W-Ub is effectively conjugated and deconjugated but the conjugates are not degraded via the ubiquitin proteasome pathways (UPP). Expression of K6W-ubiquitin in the lens and lens cells results in accumulation of intracellular aggregates and also slows cell proliferation and the differentiation program, including expression of lens specific proteins, differentiation of epithelial cells into fibers, achieving proper fiber cell morphology, and removal of nuclei. The latter is critical for transparency, but the mechanism by which cell nuclei are removed has remained an age old enigma. This was also solved by expressing K6W-Ub. p27(kip), a UPP substrate accumulates in lenses which express K6W-Ub. This precludes phosphorylation of nuclear lamin by the mitotic kinase, a prerequisite for disassembly of the nuclear membrane. Thus the nucleus remains intact and DNAseIIβ neither gains entry to the nucleus nor degrades the DNA. These results could not be obtained using chemical proteasome inhibitors that cannot be directed to specific tissues. K6W-Ub provides a novel, genetic means to study functions of the UPP because it can be targeted to specific cells and tissues. A fully functional UPP is required to execute most stages of lens differentiation, specifically removal of cell nuclei. In the absence of a functional UPP, small aggregate prone, cataractous lenses are formed.


ModuleDiscoverer: Identification of regulatory modules in protein-protein interaction networks.
The identification of disease-associated modules based on protein-protein interaction networks (PPINs) and gene expression data has provided new insights into the mechanistic nature of diverse diseases. However, their identification is hampered by the detection of protein communities within large-scale, whole-genome PPINs. A presented successful strategy detects a PPIN's community structure based on the maximal clique enumeration problem (MCE), which is a non-deterministic polynomial time-hard problem. This renders the approach computationally challenging for large PPINs implying the need for new strategies. We present ModuleDiscoverer, a novel approach for the identification of regulatory modules from PPINs and gene expression data. Following the MCE-based approach, ModuleDiscoverer uses a randomization heuristic-based approximation of the community structure. Given a PPIN of Rattus norvegicus and public gene expression data, we identify the regulatory module underlying a rodent model of non-alcoholic steatohepatitis (NASH), a severe form of non-alcoholic fatty liver disease (NAFLD). The module is validated using single-nucleotide polymorphism (SNP) data from independent genome-wide association studies and gene enrichment tests. Based on gene enrichment tests, we find that ModuleDiscoverer performs comparably to three existing module-detecting algorithms. However, only our NASH-module is significantly enriched with genes linked to NAFLD-associated SNPs. ModuleDiscoverer is available at http://www.hki-jena.de/index.php/0/2/490 (Others/ModuleDiscoverer).


Electron microprobe and synchrotron x-ray fluorescence mapping of the heterogeneous distribution of copper in high-copper vineyard soils.
The response of microorganisms to metal contamination of soils varies significantly from one investigation to another. One explanation is that metals are heterogeneously distributed at spatial scales relevant to microbes and that microoorganisms are able to avoid zones of intense contamination. This article aims to assess the microscale distribution of Cu in a vineyard soil. The spatial distribution of Cu was measured at two resolutions (0.3 mm and 20 microm) in thin sections of the surface 4 cm of undisturbed soil by electron microprobe and synchrotron X-ray microfluo-rescence spectroscopy. Bulk physicochemical analyses of Cu, pH, organic matter, texture, and mineralogy were performed. The results indicate that the Cu distribution is strongly heterogeneous at both scales of observation. Entire regions of the thin sections are virtually devoid of Cu, whereas highly localized "hotspots" have Cu signal intensities thousands of times higher than background. The distribution of Rb, or Al and Si, indicators of clay minerals, or Fe (iron (hydr)oxides), show that Cu is not preferentially associated with these mineral phases. Instead, Cu hotspots are associated with particulate organic matter. These observations suggest modification of current sampling protocols, and design of ecotoxicological experiments involving microorganisms, for contaminated soils.


MCP-1, KC-like and IL-8 as critical mediators of pathogenesis caused by Babesia canis.
Canine babesiosis caused by the intraerythrocytic protozoan parasite Babesia canis is a tick-borne disease characterized by a host response that involves both cellular and humoral immunity. This study focuses on the secretion of cytokines Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), Keratinocyte Chemotactic-like (KC-like), Interleukins (IL)-2, IL-7, IL-8, IL-10, IL-15, IL-18 and Monocyte Chemotactic Protein-1 (MCP-1) in babesiosis caused by Babesia canis upon treatment with Imizol®. We assessed time dependent changes in cytokine levels and tested whether these changes correlate with pathogenesis of the disease. Sixteen healthy dogs and 31 dogs infected with Babesia canis, of which 18 showed complications, were treated with Imizol®. One dog died during the study (3.2%). Longitudinal study was perfomed by monitoring dogs at the first day of presentation (day 1) and 6 days later (day 7). Our results show that higher MCP-1 levels on day 1 are positively associated with the occurrence of complications, (complicated vs. uncomplicated; p = 0.00016). A similar pattern was observed for KC-like on day 1 (p = 0.0326) and day 7 (p = 0.044). Moreover, babesiosis caused by B. canis produced a steady increase in IL-8 levels with a moderate to strong negative correlation with erythrocyte counts and hematocrit in uncomplicated diseased dogs only (Spearman's rank correlation coefficient rs = -0.582 and rs = -0.598 respectively). Like for MCP-1, KC-like levels also differed in complicated and uncomplicated diseased dogs on day 1 (p = 0.03236) and day 7 (p = 0.044). Furthermore, KC-like levels were strongly correlated with IL-8 levels (rs = 0.663-0.7) and non-segmented neutrophil counts (rs = 0.572-0.732) in both diseased groups. Analysis of ROC suggests the use of serum levels of MCP-1 and IL-7 as predictors of the occurrence of complications with an AUC of 0.906 and 0.896 respectively and linear combinations of MCP-1, KC-Like, IL-7 and GM-CSF with values up to AUC = 0.983. Cytokine cluster analysis presented in this study can contribute to a better understanding of the pathogenesis of babesiosis and serve as a prognostic tool for the early detection of cases with highest likelihood of developing complications. Overall, our studies show that infection by B. canis elicits a cytokine pattern that is distinct from that observed with B. rossi, and that some of the inflammatory mediators can be useful to predict complications. Our results also suggest targets for the development of novel therapeutic strategies in babesiosis caused by B. canis.