Mid to Late Life Hypertension Trends and Cerebral Small Vessel Disease in the Framingham Heart Study.
The duration and lifetime pattern of hypertension is related to risk of stroke and dementia. In turn, cerebral small vessel disease (CSVD) is the most frequent form of cerebrovascular disease underlying dementia and stroke. Thus, study of the relation of mid to late life hypertension trends with CSVD late in life will help understand hypertension's role and inform preventive efforts of CSVD consequences. We studied 1686 Framingham Heart Study Offspring cohort participants free of stroke and dementia, who were examined in mid and late life, and had available brain magnetic resonance imaging during late life. We related hypertension trends between mid and late life (normotension-normotension N-N, normotension-hypertension N-H, hypertension-hypertension H-H) to cerebral microbleeds and covert brain infarcts (CBI), overall and stratified by brain topography. We used multivariable logistic regression analyses to calculate odds ratio and 95% CIs for CSVD measures. The prevalence of CSVD in late life was 8% for cerebral microbleeds and 13% for covert brain infarcts and increased with longer hypertension exposure across all brain regions. Compared with the trend pattern of N-N, both N-H and H-H trends had higher odds of mixed cerebral microbleeds (2.71 [1.08-6.80], and 3.44 [1.39-8.60], respectively); H-H also had higher odds of any cerebral microbleeds or covert brain infarcts (1.54 [1.12-2.20]), and any covert brain infarcts (1.55 [1.08-2.20]). The burden of CSVD also increased with longer hypertension exposure. Our results highlight hypertension having a major role in subclinical CSVD, across subtypes and brain regions, and call attention to improve recognition and treatment of hypertension early in life.


SU-E-T-195: Experience in Implementing Intraoperative Radiation Therapy for Accelerated Partial Breast Irradiation Using Low-Energy X-Ray Source.
To present our preliminary experience and quality-assurance (QA) procedures in implementing intraoperative-radiation therapy (IORT) for accelerated-partial-breast irradiation (APBI) using the Axxent-system controller (Xoft Inc.) Methods: IORT was implemented in our institution utilizing a 50-keV x-ray source. APBI allows breast conserving in patients with early-stage-breast cancer by delivering radiation to the lumpectomy bed via an applicator balloon filled with water, to which a prescribed dose of 20 Gy was applied to its surface. In-situ anesthesiology, breast-surgery and radiation-oncology teams were required to achieve this procedure. Calibration of the source, including temperature and pressure corrections, was performed by the controller resulting in a calibration factor, which was manually introduced by a physicist into an in-house program providing the corrected dwell times. Evaluation of exposure measurements determined that personnel standing behind glass-rolling shields during irradiation were sufficient protection. We have successfully treated four patients to date. Surgical times averaged about two hours, QA and setup of controller took about 30 minutes, while radiation-treatment times ranged 7.8-10.4 minutes. Exposure to personnel showed a negligible dose compared to background based on the in-vivo measurements. Our measurements have shown that placement of a shielding layer (FlexiShield) on top of the drape-protected breast reduced radiation exposure to in-room personnel. In one case, treatment was automatically stopped by the controller due to a sudden undetected cooling water flow; due to the pre-treatment training, treatment was successfully resumed within a minute after manual adjustment of the flow sensor. Implementation of IORT to treat patients with APBI using low-energy x-ray source should follow the manufacturer's recommendations regarding to the acceptance and personnel training. Our experience indicates that two physicists are necessary at least at the implementation stage. Appropriate training to deal with potential problems is crucial before clinical treatment can be ready.


Effects of voxel size and iterative reconstruction parameters on the spatial resolution of 99mTc SPECT/CT.
The purpose of this study was to evaluate the effects of voxel size and iterative reconstruction parameters on the radial and tangential resolution for 99mTc SPECT as a function of radial distance from isocenter. SPECT/CT scans of eight coplanar point sources of size smaller than 1 mm3 containing high concentration 99mTc solution were acquired on a SPECT/CT system with 5/8 inch NaI(Tl) detector and low-energy, high-resolution collimator. The tomographic projection images were acquired in step-and-shoot mode for 360 views over 360° with 250,000 counts per view, a zoom of 2.67, and an image matrix of 256 × 256 pixels that resulted in a 0.9 × 0.9 × 0.9 mm3 SPECT voxel size over 230 mm field-of-view. The projection images were also rebinned to image matrices of 128 × 128 and 64 × 64 to yield SPECT voxel sizes of 1.8 × 1.8 × 1.8 and 3.6 × 3.6 × 3.6 mm3, respectively. The SPECT/CT datasets were reconstructed using the vendor-supplied iterative reconstruction software that incorporated collimator-specific resolution recovery, CT-based attenuation correction, and dual-energy window-based scatter correction using different combinations of iterations and subsets. SPECT spatial resolution was estimated as the full width at half maximum of the radial and tangential profiles through the center of each point source in reconstructed SPECT images. Both radial and tangential resolution improved with higher iterations and subsets, and with smaller voxel sizes. Both radial and tangential resolution also improved with radial distance further away from isocenter. The magnitude of variation decreased for smaller voxel sizes and for higher number of iterations and subsets. Tangential resolution was found not to be equal to the radial resolution, and the nature of the anisotropy depended on the distribution of the radionuclide and on the reconstruction parameters used. The tangential resolution converged faster than the radial resolution, with higher iterations and subsets. SPECT resolution was isotropic and independent of radial distance when reconstructed using filtered back-projection. SPECT spatial resolution and therefore quantification of SPECT uptake via partial-volume correction in clinical images were found to depend on the nature of activity distribution within the SPECT field-of-view and on the specific choice of iterative reconstruction parameters.


Mycotic aortic aneurysm infected by Clostridium septicum--a case history.
The authors describe a sixty-seven-year-old hypertensive, diabetic man with a mycotic abdominal aortic aneurysm infected with Clostridium septicum. The patient had colonic polyps but no malignant disease. They could find only one other report of a mycotic aneurysm infected with C. septicum. In that case, as in most other cases of C. septicum bacteremia, the patient had gastrointestinal cancer. Their case suggests that treatment for a clostridial infection should be considered in patients with known gastrointestinal disease, signs and symptoms of sepsis, and abdominal pain. Conversely, patients known to have a C. septicum infection should be evaluated for gastrointestinal lesions.


Serum oxidized low-density lipoprotein is inversely correlated to telomerase activity in peripheral blood mononuclear cells of haemodialysis patients.
Telomerase preserves telomeres' function and structure preventing cellular senescence. Its activity is reduced in peripheral blood mononuclear cells (PBMC) of haemodialysis (HD) patients. The purpose of this study is to investigate the potential correlation between increased oxidative stress/inflammation and telomerase activity in PBMC of HD patients. Telomerase activity was measured by PCR-ELISA in PBMC isolated from a group of 42 HD patients and 39 subjects with estimated glomerular filtration rate >or=80 mL/min (control group). Serum oxidized low-density lipoprotein (ox-LDL), tumour necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) were also measured in both groups by ELISA. Ox-LDL was negatively correlated to percentage telomerase activity in PBMC (r = -0.506, P = 0.000 in the whole group of 81 HD and normal subjects and r = -0.559, P < 0.001 in HD patients). TNF was also inversely associated with percentage telomerase activity in the whole group studied (r = -0.492, P = 0.000) while IL-10 was not. In stepwise multiple linear regression, taking into consideration the most important characteristics of the HD patients and control group, the only significant predictors for percentage telomerase activity in PBMC were ox-LDL and TNF (beta = -0.421, t = -4.083, P = 0.000 and beta = -0.381, t = -3.691, P = 0.000, respectively) while examining separately HD patients, the predictors for the same parameter were ox-LDL and HD duration (beta = -0.671, t = -4.709, P = 0.000 and beta = -0.349, t = -2.447, P = 0.023, respectively). Ox-LDL serum level is inversely correlated to telomerase activity in PBMC of HD patients. Our study proposes a new consequence of increased oxidative stress in HD patients: the premature cellular senescence potentially related to atherosclerosis through LDL oxidation.


Tolerizing DNA vaccines for autoimmune arthritis.
Current therapies for rheumatoid arthritis (RA) and other autoimmune diseases non-specifically suppress immune function, and there is great need for fundamental approaches such as antigen-specific tolerizing therapy. In this paper we describe development of antigen-specific tolerizing DNA vaccines to treat collagen-induced arthritis (CIA) in mice, and use of protein microarrays to monitor response to therapy and to identify potential additional autoimmune targets for next generation vaccines. We demonstrate that tolerizing DNA vaccines encoding type II collagen (CII) reduced the incidence and severity of CIA. Atorvastatin, a statin drug found to reduce the severity of autoimmunity, potentiated the effect of DNA vaccines encoding CII. Analysis of cytokines produced by collagen-reactive T cells derived from mice receiving tolerizing DNA encoding CII, as compared to control vaccines, revealed reduced production of the pro-inflammatory cytokines IFN-gamma and TNF-alpha. Arthritis microarray analysis demonstrated reduced spreading of autoantibody responses in mice treated with DNA encoding CII. The development of tolerizing DNA vaccines, and the use of antibody profiling to guide design of and to monitor therapeutic responses to such vaccines, represents a promising approach for the treatment of RA and other autoimmune diseases.


Neuropsychological assessment in obsessive-compulsive disorder.
Neuropsychological deficits in obsessive-compulsive disorder (OCD) have been encouraged by brain imaging studies suggesting a putative fron to- striatial biological basis of the condition. Studies of neuropsychological functions in OCD have documented deficits in several cognitive domains, particularly with regard to visuospatial abilities, executive functioning, motor speed and memory. The Aim of the present study was to assess neuropsychological profile of patients with OCD. Objectives of the study were to assess and compare the neuropsychological profile of patients with OCD and matched healthy controls. Twenty clinically stable outpatients with ICD-10 diagnosis of OCD and equal number of normal controls matched for age, education, gender and handedness were studied using a battery of neuropsychological tests. The tests consisted of verbal and performance tests of intelligence, memory, perceptual motor functions, set test and Wisconsin Card Sorting Test (WCST). On perceptual-motor functions, verbal fluency, executive functions (WCST), intelligence and memory patients with OCD did not show impairments comparable to healthy controls. An attempt to correlate the test findings with the duration of illness, stability of illness and the average drug dose was made and it was found that there was no correlation between the two. The present study does not provide evidence for a localized neuropsychological/cognitive impairment in OCD in cases that are stable for at least three months. Absence of impairments in perceptual-motor functions, verbal fluency, executive functions (WCST), intelligence, and memory does not agree with the results of other studies using these tests.


Alterations in levels of DnaK and GroEL result in diminished survival and adherence of stressed Haemophilus ducreyi.
Haemophilus ducreyi is a hemin-requiring bacterium causing the genital ulcer disease chancroid. Previously we demonstrated that the heat shock protein GroEL was immunogenic and possibly highly expressed in a mammalian host. The present study was initiated to (i) determine the relative amounts of GroEL expressed by H. ducreyi during in vitro exposure to stresses and (ii) evaluate whether a high level of GroEL is directly or indirectly required for survival and adherence of stressed H. ducreyi. Using scanning densitometry of sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles, we found that H. ducreyi expressed high basal levels of GroEL, averaging fivefold greater than in Escherichia coli. These high GroEL levels increased up to twofold upon exposure of the organism to heat shock or high levels of hydrogen peroxide and during adherence to two human genital cell lines. Furthermore, when the gene for DnaK was present on a multicopy plasmid in H. ducreyi, a 1.8-fold increase in DnaK and a 2.3-fold reduction in GroEL were seen. These results suggest that DnaK serves as a negative modulator of H. ducreyi GroEL. Subsequently we found that H. ducreyi with lower GroEL had diminished ability to survive when challenged by heat and oxidative stresses. In addition, the long, parallel chains characteristic of virulent strains of H. ducreyi were absent when GroEL was lowered, so that fewer bacterial cells adhered to the human cells. These results suggest that the unusually high basal levels of GroEL are involved, either directly or indirectly, in the survival, chaining, and adherence of H. ducreyi in the presence of the combined stresses of the host environment.


A potential application for the intracerebral injection of drugs entrapped within liposomes in the treatment of human cerebral gliomas.
After intravenous injection of therapeutic doses of bleomycin only small amounts could be measured in glioma tissue obtained at operation in patients with malignant gliomas and the drug was rapidly cleared from the blood (T1/2 = 2 hrs). Negatively charged liposomes injected through Ommaya reservoirs into the glioma tumour bed were tolerated without observable side effect. The appearance of bleomycin in the blood and urinary clearance after intracerebral injection of bleomycin entrapped within negatively charged liposomes through an Ommaya reservoir in patients with malignant gliomas was decreased as compared with levels when free bleomycin was injected. These preliminary observations suggest a role of drugs entrapped within liposomes as a depot preparation in the treatment of human cerebral gliomas.


Blood rheology adjustments in rats after a program of intermittent exposure to hypobaric hypoxia.
Intermittent hypobaric hypoxia (IHH) exposure induces a rise in hemoglobin concentration and an increase in erythrocyte mass in both rats and humans. Although this response increases blood oxygen transport capacity, paradoxically, it could impair blood flow and gas exchange because of the blood viscosity alterations associated with the rising hematocrit. In the present study, male rats were subjected to an IHH program consisting of a daily 4-h session for 5 days/week until they had completed 22 days of hypoxia exposure in a hypobaric chamber at a simulated altitude of 5000 m. Blood samples were taken at the end of the exposure period (H) and at 20 (P20) and 40 (P40) days after the end of the program and were compared to control (C) maintained at sea- level pressure. Apparent blood viscosity (eta(a)) and plasma viscosity (eta(p)) were measured in a cone-plate microviscometer. Although the hematocrit significantly increased in the H group, blood apparent viscosity did not differ among groups, ranging from 7.67 to 6.57 mPa*sec at a shear rate of 90 sec(-1). Relative blood viscosity showed a clear increase (about 27%) in H rats, mainly due to the significant decrease in plasma viscosity. This finding could be interpreted as a compensatory response, which reduced the effect of increased erythrocyte mass volume on whole-blood viscosity. Oxygen delivery index and blood oxygen potential transport capacity remained unchanged in all groups. These data indicate that the IHH program has a deep but transitory effect on red cell parameters and a moderate effect on blood rheological behavior.


Pericardial, retroperitoneal, and pleural fibrosis induced by pergolide.
Three patients with Parkinson's disease are described who developed pericardial, retroperitoneal, and pleural fibrosis associated with pergolide treatment. Surgical intervention was required in all three cases, either to reach a tissue diagnosis or for potentially life threatening complications. Symptoms emerged on average 2 years after the institution of treatment, and were sufficiently non-specific to cause significant delays in diagnosis in all cases. The erythrocyte sedimentation rate (ESR) was raised in the two patients in whom it was measured. Serosal fibrosis is a rarely reported adverse effect of pergolide treatment, although it is well described with other dopamine agonists. We suggest that patients with Parkinson's disease who receive pergolide treatment should be regularly monitored for the development of such complications.


Sex differences in referral rates of children with gender identity disorder: some hypotheses.
From 1978 through 1995, a sex ratio of 6.6:1 of boys to girls (N = 275) was observed for children referred to a specificity clinic for gender identity disorder. This article attempts to evaluate several hypotheses regarding the marked sex disparity in referral rates. The sexes did not differ on four demographic variables (age at referral, IQ, and parent's social class and marital status) and on five indices of general behavior problems on the Child Behavior Checklist; in addition, there was only equivocal evidence that boys with gender identity disorder had significantly poorer peer relations than girls with gender identity disorder. Although the percentage of boys and girls who met the complete DSM-III-R criteria for gender identity disorder was comparable, other measures of sex-typed behavior showed that the girls had more extreme cross-gender behavior than the boys. Coupled with external evidence that cross-gender behavior is less tolerated in boys than in girls by both peers and adults, it is concluded that social factors partly account for the sex difference in referral rates. Girls appear to require a higher threshold than boys for cross-gender behavior before they are referred for clinical assessment.


The Influence of β-Adrenergic Receptor Kinase-1 on Stroke-induced Immunodeficiency Syndrome.
Immunodeficiency in acute ischemic stroke (AIS) is thought to be a result of norepinephrine suppression of the lymphoid tissue. The possible differences in the distribution of lymphocytes after stroke may be due to differences in responsiveness of lymphocyte β-adrenergic receptors to their kinase (BARK-1). The objective was to quantify the influence of lymphocyte BARK-1 on stroke-induced immunodeficiency in AIS patients. A prospective clinical cohort study was conducted (N = 44). Measures included age, gender, race, risk factors for stroke, stroke severity, comorbidities, presence of infection, white blood cell counts and differential proportions, and lymphocyte BARK-1. Student t tests, effect sizes, and linear and logistic regressions were conducted to test the study objective. The study was approved by the Oregon Health & Science University Institutional Review Board. There were significant changes in all white blood cells and differential proportions and in the National Institutes of Health Stroke Scale from admission to 48 hours after onset of stroke deficits. Higher BARK-1 influenced the lower lymphocyte proportion at 48 hours, independent of age, P < .0001. Furthermore, BARK-1 also was associated with an increase in the likelihood of having sustained or stroke-induced immunodeficiency at 48 hours: odds ratio, 2.41; 95% confidence interval, 1.10-5.25; P = .027, and odds ratio, 2.79; 95% confidence interval, 1.03-7.52; P = .043, respectively. In all backward stepwise selection of factors, BARK-1 was the only factor consistently retained in the models. β-Adrenergic receptor kinase-1 has a significant quantifiable influence on lymphocyte proportion at 48 hours and on the classification of sustained stroke-induced immunodeficiency. β-Adrenergic stimulation influences immunodeficiency in AIS.


Effect of pantoprazole and Helicobacter pylori therapy on uninvestigated dyspeptic patients.
This study aimed to test the efficacy of empirical proton pump inhibitor use and Helicobacter pylori therapy for uninvestigated dyspepsia in a population with a high prevalence of H. pylori. The study had a two-stage design. In the first stage, the efficacy of 4-week pantoprazole treatment was compared with placebo in patients with uninvestigated dyspepsia. In the second stage, the efficacies of 2-week treatment with pantoprazole in H. pylori-negative patients and H. pylori eradication therapy (pantoprazole + amoxicillin + clarithromycin) in H. pylori-positive patients were compared. The primary endpoint was sufficient overall symptom relief (Global Overall Symptom score ≤2; no or minimal symptoms) at the end of treatment. In the first stage, sufficient overall symptom relief was achieved by 25.2% of patients in the pantoprazole group and 15.5% of patients in the placebo group, a difference that was not statistically significant (p=0.06). In the second stage, the rate of sufficient overall symptom relief was higher in the H. pylori therapy group than in the pantoprazole group (37.1% vs. 23.4%; p=0.02). After untreated follow-up, sufficient overall symptom relief remained significantly higher in the H. pylori therapy group than in the pantoprazole group (39.7% vs. 18%; p<0.001). Almost all patients receiving pantoprazole experienced symptom relapse after treatment. This study validated the use of a test-and-treat strategy against H. pylori in uninvestigated dyspepsia, which may be an advisable treatment approach for uninvestigated dyspeptic patients in countries with a high prevalence of H. pylori.


Anthropogenic transport of species across native ranges: unpredictable genetic and evolutionary consequences.
Human activities are responsible for the translocation of vast amounts of organisms, altering natural patterns of dispersal and gene flow. Most research to date has focused on the consequences of anthropogenic transportation of non-indigenous species within introduced ranges, with little research focusing on native species. Here, we compared genetic patterns of the sessile marine invertebrate, Ciona intestinalis, which has highly restricted dispersal capabilities. We collected individuals in a region of the species' native range where human activities that are known to facilitate the artificial spread of species are prevalent. Using microsatellite markers, we revealed highly dissimilar outcomes. First, we found low levels of genetic differentiation among sites separated by both short and large geographical distances, indicating the presence of anthropogenic transport of genotypes, and little influence of natural geographical barriers. Second, we found significant genetic differentiation in pairwise comparisons among certain sites, suggesting that other factors besides artificial transport (e.g. natural dispersal, premodern population structure) may be shaping genetic patterns. Taken together, we found dissimilar patterns of population structure in a highly urbanized region that could not be predicted by artificial transport alone. We conclude that anthropogenic activities alter genetic composition of native ranges, with unknown consequences for species' evolutionary trajectories.


Computational models of amorphous ice for accurate simulation of cryo-EM images of biological samples.
Simulations of cryo-electron microscopy (cryo-EM) images of biological samples can be used to produce test datasets to support the development of instrumentation, methods, and software, as well as to assess data acquisition and analysis strategies. To be useful, these simulations need to be based on physically realistic models which include large volumes of amorphous ice. The gold standard model for EM image simulation is a physical atom-based ice model produced using molecular dynamics simulations. Although practical for small sample volumes; for simulation of cryo-EM data from large sample volumes, this can be too computationally expensive. We have evaluated a Gaussian Random Field (GRF) ice model which is shown to be more computationally efficient for large sample volumes. The simulated EM images are compared with the gold standard atom-based ice model approach and shown to be directly comparable. Comparison with experimentally acquired data shows the Gaussian random field ice model produces realistic simulations. The software required has been implemented in the Parakeet software package and the underlying atomic models are available online for use by the wider community.


Exogenous hydrogen sulfide induces functional inhibition and cell death of cytotoxic lymphocytes subsets.
The toxic effects of exogenous hydrogen sulfide on peripheral blood lymphocytes have been investigated in detail. Hydrogen sulfide is now considered as a gasotransmitter with specific functional roles in different cell types, like neurons and vascular smooth muscle. Here we show that exogenous hydrogen sulfide induces a caspase-independent cell death of peripheral blood lymphocytes that depends on their intracellular glutathione levels, with a physiologically relevant subset specificity for CD8+ T cells and NK cells. Although lymphocyte activation does not modify their sensitivity to HS-, after 24 h exposure to hydrogen sulfide surviving lymphocyte subsets show a dramatically decreased proliferation in response to mitogens and a reduced IL-2 production. Overall, our data demonstrate that HS- reduces the cellular cytotoxic response of peripheral blood lymphocytes as well as their production of IL-2, therefore de-activating the major players of local inflammatory responses, adding new basic knowledge to the clinically well known anti-inflammatory effects of sulfur compounds.


Non-Traditional Aspects of Renal Diets: Focus on Fiber, Alkali and Vitamin K1 Intake.
Renal diets for advanced chronic kidney disease (CKD) are structured to achieve a lower protein, phosphate and sodium intake, while supplying adequate energy. The aim of this nutritional intervention is to prevent or correct signs, symptoms and complications of renal insufficiency, delaying the start of dialysis and preserving nutritional status. This paper focuses on three additional aspects of renal diets that can play an important role in the management of CKD patients: the vitamin K1 and fiber content, and the alkalizing potential. We examined the energy and nutrients composition of four types of renal diets according to their protein content: normal diet (ND, 0.8 g protein/kg body weight (bw)), low protein diet (LPD, 0.6 g protein/kg bw), vegan diet (VD, 0.7 g protein/kg bw), very low protein diet (VLPD, 0.3 g protein/kg bw). Fiber content is much higher in the VD and in the VLPD than in the ND or LPD. Vitamin K1 content seems to follow the same trend, but vitamin K2 content, which could not be investigated, might have a different pattern. The net endogenous acid production (NEAP) value decreases from the ND and LPD to the vegetarian diets, namely VD and VLPD; the same finding occurred for the potential renal acid load (PRAL). In conclusion, renal diets may provide additional benefits, and this is the case of vegetarian diets. Namely, VD and VLPD also provide high amounts of fibers and Vitamin K1, with a very low acid load. These features may have favorable effects on Vitamin K1 status, intestinal microbiota and acid-base balance. Hence, we can speculate as to the potential beneficial effects on vascular calcification and bone disease, on protein metabolism, on colonic environment and circulating levels of microbial-derived uremic toxins. In the case of vegetarian diets, attention must be paid to serum potassium levels.


Computerized assessment of complications after colorectal surgery: is it valid?
Historically, complication rates after colorectal surgery have been stratified by disease process, type of operation, or anesthesia risk derived after an intensive review of the medical record. Newer computer applications purport to shorten this process and predict the probability of postoperative complications by distinguishing them from comorbidities that are commingled on uniform discharge codes. We analyzed CaduCIS software, which uses discharge codes, to determine whether its predictions of comorbidity and complications were comparable to what was interpreted on the medical record. Two-hundred seventy patients were analyzed according to the principal and secondary diagnoses coded on discharge. Coding inaccuracies of clinical occurrences were identified by physician review of each medical record. The actual incidences of 17 common preoperative comorbidities and 11 postoperative complications were compared with those predicted by CaduCIS. The CaduCIS-predicted distribution of comorbidities was similar to the actual occurrences in 15 of 17 categories. The overall incidence of complications obtained by physician (actual) review was 47 percent, compared with 46 percent predicted by CaduCIS. However, there was a statistical difference between the CaduCIS-predicted and the actual complication rates in 5 of the 11 categories. The most common preoperative comorbidity and complication was cardiopulmonary (47 percent and 28 percent, respectively). The overall complication rate interpreted from the medical record (47 percent) was accurately predicted by CaduCIS (46 percent). Predictions of 5 of 11 individual complications were underestimated because of charting and coding inaccuracies, not because of computerized errors. Because uniform discharge coding of commingled comorbidity and complications is increasingly used to rapidly compute surgical outcomes, colon and rectal surgeons need to ensure compatibility of the actual and coded medical records.


Role of plasma exchange in ABO-incompatible kidney transplantation.
In the past, ABO incompatibility was an absolute contraindication for solid organ transplantation. However, multiple recent trials have suggested strategies for overcoming the reactions between graft antigens and recipient antibodies that cause graft rejection. In this study, we determined the usefulness of plasma exchange (PE) for removing anti-A/B antibodies that cause hyperacute/acute humoral graft rejection in patients undergoing ABO-incompatible kidney transplantation. In our study, 12 patients underwent ABO-incompatible kidney transplantation. All recipients received pre-transplantation conditioning by PE or intravenous immunoglobulin (IVIG) administration. After pre-transplantation conditioning, anti-A/B antibody titers were evaluated, and transplantation was performed when the titer was below 1:8. To assess the transplantation outcome, anti-A/B antibody titers, creatinine level, estimated glomerular filtration rate (eGFR), and proteinuria levels were measured. Anti-A/B antibody titers were below 1:8 in all patients at the time of transplantation. eGFR measured on post-transplant day 14 showed that 10 patients had immediate recovery of graft function, while 2 patients had slow recovery of graft function. Short-term outcomes of ABO-incompatible kidney transplantation (measured as creatinine levels) after reducing anti-A/B antibody titers were similar to those of ABO-compatible kidney transplantation. After transplantation, the anti-A/B antibody titers were below 1:8 in 7 patients, but the remaining 5 patients required post-transplantation PE and IVIG treatment to prevent antigen-antibody reactions. With the increasing demand for kidney donations, interest in overcoming the ABO incompatibility barrier has increased. PE may be an important breakthrough in increasing the availability of kidneys for transplantation.


Salmonella spp., Vibrio spp., Clostridium perfringens, and Plesiomonas shigelloides in marine and freshwater invertebrates from coastal California ecosystems.
The coastal ecosystems of California are highly utilized by humans and animals, but the ecology of fecal bacteria at the land-sea interface is not well understood. This study evaluated the distribution of potentially pathogenic bacteria in invertebrates from linked marine, estuarine, and freshwater ecosystems in central California. A variety of filter-feeding clams, mussels, worms, and crab tissues were selectively cultured for Salmonella spp., Campylobacter spp., Escherichia coli-O157, Clostridium perfringens, Plesiomonas shigelloides, and Vibrio spp. A longitudinal study assessed environmental risk factors for detecting these bacterial species in sentinel mussel batches. Putative risk factors included mussel collection near higher risk areas for livestock or human sewage exposure, adjacent human population density, season, recent precipitation, water temperature, water type, bivalve type, and freshwater outflow exposure. Bacteria detected in invertebrates included Salmonella spp., C. perfringens, P. shigelloides, Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio alginolyticus. Overall, 80% of mussel batches were culture positive for at least one of the bacterial species, although the pathogens Campylobacter, E. coli-O157, and Salmonella were not detected. Many of the same bacterial species were also cultured from upstream estuarine and riverine invertebrates. Exposure to human sewage sources, recent precipitation, and water temperature were significant risk factors for bacterial detection in sentinel mussel batches. These findings are consistent with the hypothesis that filter-feeding invertebrates along the coast concentrate fecal bacteria flowing from land to sea and show that the relationships between anthropogenic effects on coastal ecosystems and the environmental niches of fecal bacteria are complex and dynamic.


Tar, CO and delta 9THC delivery from the 1st and 2nd halves of a marijuana cigarette.
Previous in vitro studies suggest that, with successive puffs from a marijuana cigarette, delta-9-THC becomes concentrated in the remaining uncombusted portion of the cigarette. These observations are consistent with the common practice of smoking marijuana cigarettes to a smaller butt length than that to which tobacco cigarettes are smoked. The purpose of the present study was to compare the delivery of delta-9-THC, as well as total insoluble smoke particulates (tar) and carbon monoxide, from the distal ("first") versus the proximal ("second") halves of a standard marijuana cigarette during "natural" smoking of marijuana. On 4 separate days, ten habitual marijuana users smoked nearly all or approximately 1/2 of a standard marijuana cigarette (83 mm length; 800-900 mg; 1.24% THC), as follows: day 1, "whole" cigarette (60 mm smoked, leaving a 23-mm butt); day 2, "first" half (first 30 mm); day 3, "second" half (second 30 mm) after the "first" half was presmoked with a syringe; and day 4, "second" half after the "first" half was excised. A previously described smoking apparatus (20) was used for measurement of puff volume and inhaled tar. Puff volume and number were allowed to vary spontaneously (provided that the specified length of cigarette was consumed), while inhaled volume (1.5 liters), breathholding time (14 s) and interpuff interval (30 s) were held constant. Blood samples were withdrawn prior to smoking and serially after completion of smoking for analysis of blood carboxyhemoglobin (COHb) and serum delta-9-THC. Heart rate was measured before and 5 min after smoking. Subjects rated their level of "high" 20 min after completion of smoking.(ABSTRACT TRUNCATED AT 250 WORDS)


Oil-in-microgel strategy for enzymatic-triggered release of hydrophobic drugs.
Polymer microgels have received considerable attention due to their great potential in the biomedical field as drug delivery systems. Hyaluronic acid (HA) is a naturally occurring glycosaminoglycan composed of N-acetyl-d-glucosamine and d-glucuronic acid. This polymer is biodegradable, nontoxic, and can be chemically modified. In this work, a co-flow microfluidic strategy for the preparation of biodegradable HA microgels encapsulating hydrophobic drugs is presented. The approach relies on: (i) generation of a primary oil-in-water (O/W) nanoemulsion by the ultrasonication method, (ii) formation of a double oil-in-water-in-oil emulsion (O/W/O) using microfluidics, and (iii) cross-linking of microgels by photopolymerization of HA precursors modified with methacrylate groups (HA-MA) present in the aqueous phase of the droplets. The procedure is used for the encapsulation and controlled release of progesterone. Degradability and encapsulation/release studies in PBS buffer at 37°C in presence of different concentrations of hyaluronidase are performed. It is demonstrated that enzymatic degradation can be used to trigger the release of progesterone from microgels. This method provides precise control of the release system and can be applied for the encapsulation and controlled release of different types of hydrophobic drugs.


Firing up the cold tumors by targeting Vps34.
Cancer immunotherapy based on anti-PD-1/PD-L1 blockade is particularly effective in responding to patients with hot tumors. These tumors are characterized by the accumulation of proinflammatory cytokines and T cell infiltration. In our recent report published in Science Advances, we demonstrate that targeting the autophagy-related protein Vps34 switched cold immune desert tumors into hot inflamed immune-infiltrated tumors and enhanced the efficacy of anti-PD-1/PD-L1. Our study provides the preclinical rationale to set up combination immunotherapy clinical trials using selective Vps34 inhibitors and immune checkpoint blockers in melanoma and CRC.


Salivary response to food in humans and its effect on gastric acid secretion.
The purpose of these studies was to determine the amount of saliva secreted in response to food in humans and to evaluate the effect of saliva on postprandial gastric acid secretion. Subjects chewed and spat out an appetizing steak and french-fried potato meal (modified sham feeding), with the increase in the weight of the meal during sham feeding taken to represent salivary secretion. Mean salivary flow rose from basal rates of 37.1 ml/h to 167.3 ml/h during modified sham feeding (P less than 0.001). Chewing gum increased salivary secretion to approximately the same extent as modified sham feeding, whereas chewing on plastic tubing caused a much smaller increase in salivary flow. Intravenous infusion of bethanechol (50 micrograms . kg-1 . h-1) was approximately half as potent as modified sham feeding or gum chewing in stimulating salivary flow. The salivary response to sham feeding was completely blocked by 15 micrograms/kg intravenous atropine. Salivary secretion increased approximately 20 ml/h when a 700-ml amino acid meal was infused directly into the stomach (P less than 0.01), whereas gastric distension with 700 ml saline had no effect. These findings suggested that food in the stomach or upper small intestine may activate a reflex or release a hormone into the circulation that augments salivary flow. Although intravenous gastrin-17 infusion had no effect on salivary flow, somatostatin-14 infusion increased salivary flow 15 ml/h (P less than 0.02). Saliva contained approximately 2,000 pg/ml immunoreactive urogastrone, an inhibitor of acid secretion when administered parenterally.(ABSTRACT TRUNCATED AT 250 WORDS)


Structural basis for substrate delivery by acyl carrier protein in the yeast fatty acid synthase.
In the multifunctional fungal fatty acid synthase (FAS), the acyl carrier protein (ACP) domain shuttles reaction intermediates covalently attached to its prosthetic phosphopantetheine group between the different enzymatic centers of the reaction cycle. Here, we report the structure of the Saccharomyces cerevisiae FAS determined at 3.1 angstrom resolution with its ACP stalled at the active site of ketoacyl synthase. The ACP contacts the base of the reaction chamber through conserved, charge-complementary surfaces, which optimally position the ACP toward the catalytic cleft of ketoacyl synthase. The conformation of the prosthetic group suggests a switchblade mechanism for acyl chain delivery to the active site of the enzyme.


One-year symptom trajectories in patients with stable schizophrenia maintained on antipsychotics versus placebo: meta-analysis.
BackgroundAs definitions of relapse differ substantially between studies, in investigations involving data aggregation, total scores on clinical rating scales provide a more generalisable outcome.AimsTo compare total symptom trajectories for antipsychotic versus placebo treatment over a 1-year period of maintenance treatment in schizophrenia.MethodRandomised controlled trials with antipsychotic and placebo treatment arms in patients with stable schizophrenia that reported Positive and Negative Syndrome Scale and Brief Psychiatric Rating Scale total scores at more than one time point were included. Meta-regression analyses were employed using a mixed model.ResultsA total of 11 studies involving 2826 patients were included. Meta-regression analyses revealed significant interactions between group and time (PS<0.0001); both standardised total scores and per cent score changes remained almost unchanged in patients continuing antipsychotic treatment, whereas symptoms continuously worsened over time in those switching to placebo treatment.ConclusionsWhen considering long-term antipsychotic treatment of schizophrenia, clinicians must balance symptomatic and functional outcomes.


Characterization of antibodies to the glycosyl-phosphatidylinositol membrane anchors of mammalian proteins.
Two polyclonal antisera were raised in rabbits to the phospholipase C-solubilized forms of pig renal dipeptidase (EC 184.108.40.206) and pig aminopeptidase P (EC 18.104.22.168). These antisera were purified and shown to cross-react with other glycosyl-phosphatidylinositol (G-PI)-anchored proteins isolated from pig, human and trypanosomes. The epitopes involved in this cross-reactivity were characterized by Western-blot analysis after mild acid or nitrous acid treatment of the G-PI-anchored proteins and by a competitive e.l.i.s.a. with other G-PI-anchored proteins and individual components of the anchor structure. These studies revealed that the primary epitope for both antisera is the inositol 1.2-(cyclic)monophosphate that is formed on phospholipase C cleavage of the intact G-PI anchor. Other minor epitopes, such as phosphoethanolamine, probably involve side-chain modifications to the core anchor structure that may be species-specific.


Minimally Manipulated Bone Marrow Concentrate Compared with Microfracture Treatment of Full-Thickness Chondral Defects: A One-Year Study in an Equine Model.
Microfracture is commonly performed for cartilage repair but usually results in fibrocartilage. Microfracture augmented by autologous bone marrow concentrate (BMC) was previously shown to yield structurally superior cartilage repairs in an equine model compared with microfracture alone. The current study was performed to test the hypothesis that autologous BMC without concomitant microfracture improves cartilage repair compared with microfracture alone. Autologous sternal bone marrow aspirate (BMA) was concentrated using a commercial system. Cells from BMC were evaluated for chondrogenic potential in vitro and in vivo. Bilateral full-thickness chondral defects (15-mm diameter) were created on the midlateral trochlear ridge in 8 horses. Paired defects were randomly assigned to treatment with BMC without concomitant microfracture, or to microfracture alone. The repairs were evaluated at 1 year by in vitro assessment, arthroscopy, morphological magnetic resonance imaging (MRI), quantitative T2-weighted and ultrashort echo time enhanced T2* (UTE-T2*) MRI mapping, and histological assessment. Culture-expanded but not freshly isolated cells from BMA and BMC underwent cartilage differentiation in vitro. In vivo, cartilage repairs in both groups were fibrous to fibrocartilaginous at 1 year of follow-up, with no differences observed between BMC and microfracture by arthroscopy, T2 and UTE-T2* MRI values, and histological assessment (p > 0.05). Morphological MRI showed subchondral bone changes not observed by arthroscopy and improved overall outcomes for the BMC repairs (p = 0.03). Differences in repair tissue UTE-T2* texture features were observed between the treatment groups (p < 0.05). When BMC was applied directly to critical-sized, full-thickness chondral defects in an equine model, the cartilage repair results were similar to those of microfracture. Our data suggest that, given the few mesenchymal stem cells in minimally manipulated BMC, other mechanisms such as paracrine, anti-inflammatory, or immunomodulatory effects may have been responsible for tissue regeneration in a previous study in which BMC was applied to microfractured repairs. While our conclusions are limited by small numbers, the better MRI outcomes for the BMC repairs may have been related to reduced surgical trauma to the subchondral bone. MRI provides important information on chondral defect subsurface repair organization and subchondral bone structure that is not well assessed by arthroscopy.


Elevated water temperature and carbon dioxide concentration increase the growth of a keystone echinoderm.
Anthropogenic climate change poses a serious threat to biodiversity. In marine environments, multiple climate variables, including temperature and CO(2) concentration ([CO(2)]), are changing simultaneously. Although temperature has well-documented ecological effects, and many heavily calcified marine organisms experience reduced growth with increased [CO(2)], little is known about the combined effects of temperature and [CO(2)], particularly on species that are less dependent on calcified shells or skeletons. We manipulated water temperature and [CO(2)] to determine the effects on the sea star Pisaster ochraceus, a keystone predator. We found that sea star growth and feeding rates increased with water temperature from 5 degrees C to 21 degrees C. A doubling of current [CO(2)] also increased growth rates both with and without a concurrent temperature increase from 12 degrees C to 15 degrees C. Increased [CO(2)] also had a positive but nonsignificant effect on sea star feeding rates, suggesting [CO(2)] may be acting directly at the physiological level to increase growth rates. As in past studies of other marine invertebrates, increased [CO(2)] reduced the relative calcified mass in sea stars, although this effect was observed only at the lower experimental temperature. The positive relationship between growth and [CO(2)] found here contrasts with previous studies, most of which have shown negative effects of [CO(2)] on marine species, particularly those that are more heavily calcified than P. ochraceus. Our findings demonstrate that increased [CO(2)] will not have direct negative effects on all marine invertebrates, suggesting that predictions of biotic responses to climate change should consider how different types of organisms will respond to changing climatic variables.


Cardiovascular disease-related hospital admissions of patients with inflammatory arthritis.
Patients with inflammatory arthritis (IA) have an increased risk of cardiovascular diseases (CVD), suggesting a high rate of CVD-related hospitalizations, but data on this topic are limited. Our study addressed hospital admissions for CVD in a primary care-based population of patients with IA and controls. All newly diagnosed patients with IA between 2001 and 2010 were selected from electronic medical records of the Netherlands Institute for Health Services Research Primary Care database, representing a national network of general practices. Two control patients matched for age, sex, and practice were selected for each patient with IA. Hospital admission data for all patients was retrieved from the Dutch Hospital Data. There were 2615 patients with IA and 5555 controls included in our study. CVD-related hospital admissions were observed more frequently among patients with IA as compared with control patients: 48% versus 36% (p < 0.001) in a followup period of 4 years. Patients with IA were more often hospitalized because of ischemic heart disease (OR 1.7, 95% CI 1.2-2.2) and for day-care admission because of cerebrovascular disease (OR 2.2, 95% CI 1.0-4.9). Increased hospital admission rates confirm the higher CVD burden among patients with IA compared with controls, and underscore the need for proper CVD risk management in patients with IA.


Men in Occupational Therapy: Issues, Factors, and Perceptions.
Relatively few men are occupational therapy practitioners in the United States. Little research has examined issues germane to men working in the profession. The objective of this study was to investigate perceptions, issues, and factors surrounding male occupational therapy practitioners. A survey link was emailed to 9,467 licensed occupational therapy practitioners in Ohio; 699 participants responded, yielding a response rate of 7.66%. Male and female participants provided both similar and significantly different responses to the survey questions. Differences generally involved responses to questions on personal finances and client education. Themes were identified from the open-ended questions regarding gender stereotypes. Although male and female practitioners differed in opinion on several issues and perceptions related to the profession, in general they were happy with their career choice. Specifically, participants expressed that they enjoyed their job, contributed to society, and had a sense of job security, regardless of gender.


A scoping review on health economics in neurosurgery for acute spine trauma.
OBJECTIVE Acute spine trauma (AST) has a relatively low incidence, but it often results in substantial individual impairments and societal economic burden resulting from the associated disability. Given the key role of neurosurgeons in the decision-making regarding operative management of individuals with AST, the authors performed a systematic search with scoping synthesis of relevant literature to review current knowledge regarding the economic burden of AST. METHODS This systematic review with scoping synthesis included original articles reporting cost-effectiveness, cost-utility, cost-benefit, cost-minimization, cost-comparison, and economic analyses related to surgical management of AST, whereby AST is defined as trauma to the spine that may result in spinal cord injury with motor, sensory, and/or autonomic impairment. The initial literature search was carried out using MEDLINE, EMBASE, CINAHL, CCTR, and PubMed. All original articles captured in the literature search and published from 1946 to September 27, 2017, were included. Search terms used were the following: (cost analysis, cost effectiveness, cost benefit, economic evaluation or economic impact) AND (spine or spinal cord) AND (surgery or surgical). RESULTS The literature search captured 5770 titles, of which 11 original studies met the inclusion/exclusion criteria. These 11 studies included 4 cost-utility analyses, 5 cost analyses that compared the cost of intervention with a comparator, and 2 studies examining direct costs without a comparator. There are a few potentially cost-saving strategies in the neurosurgical management of individuals with AST, including 1) early surgical spinal cord decompression for acute traumatic cervical spinal cord injury (or traumatic thoracolumbar fractures, traumatic cervical fractures); 2) surgical treatment of the elderly with type-II odontoid fractures, which is more costly but more effective than the nonoperative approach among individuals with age at AST between 65 and 84 years; 3) surgical treatment of traumatic thoracolumbar spine fractures, which is implicated in greater direct costs but lower general-practitioner visit costs, private expenditures, and absenteeism costs than nonsurgical management; and 4) removal of pedicle screws 1-2 years after posterior instrumented fusion for individuals with thoracolumbar burst fractures, which is more cost-effective than retaining the pedicle screws. CONCLUSIONS This scoping synthesis underscores a number of potentially cost-saving opportunities for neurosurgeons when managing patients with AST. There are significant knowledge gaps regarding the potential economic impact of therapeutic choices for AST that are commonly used by neurosurgeons.


Young infants' neural processing of objects is affected by eye gaze direction and emotional expression.
Eye gaze is an important social cue which is used to determine another person's focus of attention and intention to communicate. In combination with a fearful facial expression eye gaze can also signal threat in the environment. The ability to detect and understand others' social signals is essential in order to avoid danger and enable social evaluation. It has been a matter of debate when infants are able to use gaze cues and emotional facial expressions in reference to external objects. Here we demonstrate that by 3 months of age the infant brain differentially responds to objects as a function of how other people are reacting to them. Using event-related electrical brain potentials (ERPs), we show that an indicator of infants' attention is enhanced by an adult's expression of fear toward an unfamiliar object. The infant brain showed an increased Negative central (Nc) component toward objects that had been previously cued by an adult's eye gaze and frightened facial expression. Our results further suggest that infants' sensitivity cannot be due to a general arousal elicited by a frightened face with eye gaze directed at an object. The neural attention system of 3 month old infants is sensitive to an adult's eye gaze direction in combination with a fearful expression. This early capacity may lay the foundation for the development of more sophisticated social skills such as social referencing, language, and theory of mind.


Structure-based tailoring of compound libraries for high-throughput screening: discovery of novel EphB4 kinase inhibitors.
High-throughput docking is a computational tool frequently used to discover small-molecule inhibitors of enzymes or receptors of known three-dimensional structure. Because of the large number of molecules in chemical libraries, automatic procedures to prune multimillion compound collections are useful for high-throughput docking and necessary for in vitro screening. Here, we propose an anchor-based library tailoring approach (termed ALTA) to focus a chemical library by docking and prioritizing molecular fragments according to their binding energy which includes continuum electrostatics solvation. In principle, ALTA does not require prior knowledge of known inhibitors, but receptor-based pharmacophore information (hydrogen bonds with the hinge region) is additionally used here to identify molecules with optimal anchor fragments for the ATP-binding site of the EphB4 receptor tyrosine kinase. The 21,418 molecules of the focused library (from an initial collection of about 730,000) are docked into EphB4 and ranked by force-field-based energy including electrostatic solvation. Among the 43 compounds tested in vitro, eight molecules originating from two different anchors show low-micromolar activity in a fluorescence-based enzymatic assay. Four of them are active in a cell-based assay and are potential anti-angiogenic compounds.


Care home versus hospital and own home environments for rehabilitation of older people.
Rehabilitation for older people has acquired an increasingly important profile for both policy-makers and service providers within health and social care agencies. This growing demand for rehabilitation services has generated an increased interest in the use of alternative care environments, for example care home environments, for older persons' rehabilitation. At a time when there is pressure for policy decision-makers and service providers to explore the use of such care settings for the provision of rehabilitation for older people, there appears limited evidence on which to base decisions. The objective of this review is to compare the effects of care home environments (e.g. nursing home, residential care home and nursing facilities) versus hospital environments and own home environments in the rehabilitation of older people. The following databases were searched. The Cochrane Effective Practice and Organisation of Care Specialised Register, the Cochrane Rehabilitation Specialist Register; Cochrane Controlled Trials Register (CCTR); MEDLINE (1966-2000); EMBASE (1980-2000), Cumulative Index to Nursing and Allied Health Literature (CINAHL) (1982-2000): Science Citation Index (1982-2000); Social Science Citation Index (1982-2000); Best Evidence (1991-2000); HMIC (1979-2000); PsycINFO(1967-2000); ASSIA (1987-2000); Ageline (1978-2000); AgeInfo (1971-2000); Sociological Abstracts (1963-2000); System for Information on Grey Literature (SIGLE) (1980-2000); UK National Research Registers Project Database( Issue 1 2001); Architecture Publication Index (1977-2000). The following Journals were hand searched: Disability and Rehabilitation (1992-2000); Disability and Society (1986-2000); Archives of Physical Medicine and Rehabilitation (1985-2000); Journal of the American Geriatric Society (1980-2000); International Journal of Rehabilitation Research (1980-2000); American Journal of Physical Medicine and Rehabilitation (1980-2000) and: Clinical Rehabilitation (1992-2000). The reviewers also consulted subject area experts and obtained full text review articles and forward tracked any references from these sources. Randomised controlled trials (RCTs), controlled clinical trials (CCTs), controlled before and after studies (CBAs) and interrupted time series (ITS) that compared rehabilitation outcomes for persons 60 years or older who received rehabilitation whilst residing in a care home with those for persons 60 years or older who received rehabilitation in hospital or own home environments. Primary outcomes included functional outcomes using activities of daily living measurement (both personal and instrumental). Secondary outcomes included subjective health status; quality of life measures; return to place of usual residency; all cause mortality; adverse effects; readmission to an acute care facility; patient and carer satisfaction; number of days in facility and number of days receiving rehabilitation. One reviewer (DW) completed the initial search and identified potential papers for inclusion. Abstracts for these papers were independently scrutinised by two reviewers (DW/MS) to assess their eligibility. Full text versions of potentially eligible papers were independently assessed by two reviewers (DW/MS). Papers that fulfilled the comparison inclusion criteria were then independently scrutinised by all reviewers to assess whether they met EPOC methodological criteria for inclusion. The total yield from the initial search strategy was 19,457. A total of 1,247 abstracts were independently scrutinised by two reviewers (DW/MS) to assess their eligibility. Full text papers for 99 studies were obtained to assess if they fulfilled the review's comparison inclusion criteria. This process resulted in 12 papers being assessed further for methodological validity. However, none of these studies met the inclusion criteria. There is insufficient evidence to compare the effects of care home environments, hospital environments and own home environments on older persons rehabilitation outcomes. Although the authors acknowledge that absence of effect is not no effect. There are three main reasons; the first is that the description and specification of the environment is often not clear; secondly, the components of the rehabilitation system within the given environments are not adequately specified and; thirdly, when the components are clearly specified they demonstrate that the control and intervention sites are not comparable with respect to the methodological criteria specified by Cochrane EPOC group (Cochrane 1998). The combined effect of these factors resulted in the comparability between intervention and control groups being very weak. For example, there were differences in the services provided in the intervention and control arms, due possibly to differences in dominant remuneration systems, nature of the rehabilitation transformation, patient characteristics, skill mix and academic status of the care environment.


Evaluation of a novel automated volumetric flow cytometer for absolute CD4+ T lymphocyte quantitation.
Bead-based single platform cytometry technology (SPT) is the gold standard when performing CD4 absolute counting. However, it presents drawbacks as precision depends on various critical steps (for example, pipetting methodology, overtime stability of beads, stability of fluidics, regular recalibration…) and thus requires skilled operators. The fully automated volumetric SPT AQUIOS CL (Beckman Coulter) has recently emerged as an alternative with no need of beads. It may help improving results standardisation and fulfilling requirements for certification (ISO 15189). In this study, we assessed SPT AQUIOS CL performances in accordance to requirements for ISO 15189 accreditation. We evaluated repeatability and reproducibility (precision), bias (trueness), uncertainty (total error), range limits (linearity, quantification, detection limits), and inter-reagent/inter-sample contaminations in enumerating CD4+ T-cells. Concomitantly, we compared AQUIOS CL CD4+ T-cell values with the results obtained with our routine bead-based SPT (that is, FC500 Beckman Coulter, bead-based SPT), on blood samples from 148 patients representative of clinical laboratory routine workload. Every result (repeatability, reproducibility, trueness, total error) was below the acceptable thresholds proposed in international recommendations. Contamination results and range limits (linearity, quantification, and detection limits) were all found perfectly suitable to routine analysis. The comparison between AQUIOS CL and FC500 exhibited excellent correlation and agreement (Pearson R = 0.99, P < 0.001; Lin's concordance correlation coefficient: Lin ρc = 0.991, Cb = 0.999), and Bland-Altman analysis did not reveal any systematic error. Our results demonstrate that, upon subsequent validation in more routine conditions, the AQUIOS CL could be a suitable tool for clinical flow cytometry laboratories facing accreditation process. © 2016 International Clinical Cytometry Society.


Presentation, management, and outcome of out of hospital cardiopulmonary arrest: comparison by underlying aetiology.
To describe and compare presentation, management, and survival by aetiology of cardiopulmonary arrest. A retrospective cohort study was undertaken of all 21 175 first out of hospital cardiopulmonary arrests in Scotland between May 1991 and March 1998. Discharge alive from hospital. Presumed cardiac disease accounted for 17 451 cases (82%), other internal aetiologies for 1814 (9%), and external aetiologies for 1910 (9%). Arrests caused by presumed cardiac disease had a better risk profile in terms of presence of a witness, bystander cardiopulmonary resuscitation, call-response interval, and use of defibrillation; 1265 (7%) of those who arrested from presumed cardiac disease were discharged alive, compared with only 77 (2%) of those with non-cardiac disorders (p < 0.001). Among those defibrillated, call-response interval was associated with survival following arrests from both presumed cardiac and non-cardiac causes (p < 0.001). Out of hospital cardiopulmonary arrests from non-cardiac causes were associated with worse crude survival than arrests from cardiac causes. Improvements in call-response interval and basic life support skills in the community would improve survival irrespective of the aetiology and should therefore be encouraged.


Altered Toll-like receptor 2-mediated endotoxin tolerance is related to diminished interferon beta production.
Induction of endotoxin tolerance leads to a reduced inflammatory response after repeated challenge by LPS and is important for resolution of inflammation and prevention of tissue damage. Enterobacterial LPS is recognized by the TLR4 signaling complex, whereas LPS of some non-enterobacterial organisms is capable of signaling independently of TLR4 utilizing TLR2-mediated signal transduction instead. In this study we report that Porphyromonas gingivalis LPS, a TLR2 agonist, fails to induce a fully endotoxin tolerant state in a human monocytic cell line (THP-1) and mouse bone marrow-derived macrophages. In contrast to significantly decreased production of human IL-8 and TNF-α and, in mice, keratinocyte-derived cytokine (KC), macrophage inflammatory protein-2 (MIP-2), and TNF-α after repeated challenge with Escherichia coli LPS, cells repeatedly exposed to P. gingivalis LPS responded by producing less TNF-α but sustained elevated secretion of IL-8, KC, and MIP-2. Furthermore, in endotoxin-tolerant cells, production of IL-8 is controlled at the signaling level and correlates well with NF-κB activation, whereas TNF-α expression is blocked at the gene transcription level. Interferon β plays an important role in attenuation of chemokine expression in endotoxin-tolerized cells as shown in interferon regulatory factor-3 knock-out mice. In addition, human gingival fibroblasts, commonly known not to display LPS tolerance, were found to be tolerant to repeated challenge by LPS if pretreated with interferon β. The data suggest that the inability of the LPS-TLR2 complex to induce full endotoxin tolerance in monocytes/macrophages is related to diminished production of interferon β and may partly explain the involvement of these LPS isoforms in the pathogenesis of chronic inflammatory diseases.


Sensory profile, ferritin and zinc levels in preschool-aged children with symptoms of attention deficit hyperactivity disorder.
Attention Deficit Hyperactivity Disorder (ADHD) is one of the most common neurodevelopmental disorders and has a big impact on the well-being of children. The disorder can lead to noticeable functional limitations for children and bio-ecological factors also contribute to symptoms of ADHD. We aimed to investigate the associations between ADHD symptoms and some related bio-ecological factors including serum ferritin, zinc levels and sensory processing in preschool-aged children. Twenty-two children who had been referred to the division of Developmental Pediatrics because of ADHD symptoms and 22 participants from the general pediatric outpatient clinics were included in the study. The symptoms of ADHD were evaluated with Conners` Parent Rating Scale-Revised Short form. Complete blood count, serum ferritin and zinc levels were also evaluated. A blind occupational therapist implemented sensory processing measurements. The characteristics of each participant such as prematurity, perinatal complications, developmental practices and sociodemographic data were also considered. Sensory processing measurement analysis revealed that all Sensory Profile scores were significantly lower in the children with ADHD symptoms compared to the control group indicating that the child shows the behavior more than desired. The low level of zinc (p=0.026, OR=6.153, 95% CI= 1.247-30.362) and the presence of perinatal complications (p=0.045, OR=10.864, 95% CI=1.059-111.499) increased the risk of ADHD symptoms. We could not find an association for ferritin levels in our study. The evaluation of zinc level and sensory profile parallel to other strategies can be recommended during the management of ADHD symptoms in preschool children.


Specific hypomethylated CpGs at the IGF2 locus act as an epigenetic biomarker for familial adenomatous polyposis colorectal cancer.
The identification of specific biomarkers for colorectal cancer is of primary importance for early diagnosis. The aim of this study was to evaluate if methylation changes at the IGF2/H19 locus could be predictive for individuals at high risk for developing sporadic or hereditary colorectal cancer. Quantitative methylation analysis using pyrosequencing was performed on three differentially methylated regions (DMRs): IGF2 DMR0 and DMR2 and the H19 DMR in DNA samples from sporadic colorectal cancer (n = 26), familial adenomatous polyposis (n = 35) and hereditary nonpolyposis colorectal cancer (n = 19) patients. We report in this article for the first time, that in sporadic colorectal cancer tumor DNA both the IGF2 DMR0 and DMR2 are hypomethylated, while the H19 DMR retains its monoallelic methylation pattern. In lymphocyte DNA, a striking hypomethylation of nine contiguous correlated CpGs was found in the IGF2 DMR2 but only in familial adenomatous polyposis patients. Methylation alterations at the IGF2 locus are more extensive than previously reported and DMR2 hypomethylation in lymphocyte DNA might be a specific epigenetic biomarker for familial adenomatous polyposis patients.


The leptomeningeal "ivy sign" on fluid-attenuated inversion recovery MR imaging in Moyamoya disease: a sign of decreased cerebral vascular reserve?
Moyamoya disease is an idiopathic occlusive cerebrovascular disorder with abnormal microvascular proliferation. We investigated the clinical utility of leptomeningeal high signal intensity (ivy sign) sometimes seen on fluid-attenuated inversion recovery images in Moyamoya disease. We examined the relationship between the degree of the ivy sign and the severity of the ischemic symptoms in 96 hemispheres of 48 patients with Moyamoya disease. We classified each cerebral hemisphere into 4 regions from anterior to posterior. In 192 regions of 24 patients, we examined the relationship between the degree of the ivy sign and findings of single-photon emission CT, including the resting cerebral blood flow (CBF) and cerebral vascular reserve (CVR). The degree of the ivy sign showed a significant positive relationship with the severity of the ischemic symptoms (P < .001). Of the 4 regions, the ivy sign was most frequently and prominently seen in the anterior part of the middle cerebral artery region. The degree of the ivy sign showed a negative relationship with the resting CBF (P < .0034) and a more prominent negative relationship with the CVR (P < .001). The leptomeningeal ivy sign indicates decreased CVR in Moyamoya disease.


Chronic inflammation induced by organic dust and related metabolic cardiovascular disease risk factors.
Chronic inflammation is now considered a risk factor for cardiovascular diseases. Exposure to organic dust induces an inflammatory response. This study was done to verify whether inflammation caused by exposure to organic dust increases the metabolic risk factors for cardiovascular diseases. Thirty-six nonsmoking men who worked in a swine confinement building and 35 unexposed matched controls were studied. Each person was evaluated for inflammatory markers, including white blood cell counts, cell-bound (CD11b, CD18, CD31, CD62L) and circulating soluble adhesion molecule levels (sICAM-1, sPECAM-1, sL, sE, and sP selectins), serum CRP (C-reactive protein), fibrinogen, and interleukin-6 (IL-6). Cardiovascular risk factors [the serum lipid profile, apoprotein B (Apo B)] and insulin levels were also assessed. The groups were similar with respect to age, physical characteristics, and blood cell counts. The expression of adhesion molecules (P-values <0.01) and serum levels of sL-selectin (P<0.0001) were higher for the workers than for the controls, while neutrophils and interleukin-6 were slightly higher for the workers (P=0.05). No differences in CRP, fibrinogen, lipid profile, Apo B, or insulin levels were observed. Exposure to contaminated organic dust induces a chronic inflammation that is not associated with increased metabolic or acute-phase cardiovascular risk factors; this finding suggests that chronic inflammation per se may not be a cardiovascular risk factor in this group of pig farmers.


Thermo-molecular orientation effects in fluids of dipolar dumbbells.
We use molecular dynamics simulations in applied thermal gradients to study thermomolecular orientation (TMO) of size-asymmetric dipolar dumbbells with different molecular dipole moments. We find that the direction of the TMO is the same as in apolar dumbbells of the same size, i.e. the smaller atom in the dumbbell tends to orient towards the colder temperature. The ratio of the electrical polarization to the magnitude of the thermal gradient does not vary much with the magnitude of the molecular dipole moment. We also investigate a novel second order TMO that persists even in size-symmetric dipolar dumbbells where molecules have a slight tendency to orient perpendicular to the gradient except very close to the hot region, where (anti-)parallel orientations are preferred. Finally, we investigate rotational correlation functions and characteristic rotational times in these systems in an attempt to model possible spectroscopic signatures of TMO in experiments. Although we cannot detect any difference in integrated rotational times between equilibrium simulations and simulations in a thermal gradient, more careful modelling of the anisotropic rotational dynamics in the thermal gradient may be more successful.


Multicentre surveillance of prevalence of the 23S rRNA A2058G and A2059G point mutations and molecular subtypes of Treponema pallidum in Taiwan, 2009-2013.
Resistance mutations A2058G and A2059G, within the 23S rRNA gene of Treponema pallidum, have been reported to cause treatment failures in patients receiving azithromycin for syphilis. Genotyping of T. pallidum strains sequentially isolated from patients with recurrent syphilis is rarely performed. From September 2009 to August 2013, we collected 658 clinical specimens from 375 patients who presented with syphilis for genotyping to examine the number of 60-bp repeats in the acidic repeat protein (arp) gene, T. pallidum repeat (tpr) polymorphism, and tp0548 gene, and to detect A2058G and A2059G point mutations by restriction fragment length polymorphism. Treponemal DNA was identified in 45.2% (n = 298) of the specimens that were collected from 216 (57.6%) patients; 268 (40.7%) specimens tested positive for the 23S rRNA gene, and were examined for macrolide resistance. Two isolates (0.7%) harboured the A2058G mutation, and no A2059G mutation was identified. A total of 14 strains of T. pallidum were identified, with 14f/f (57.5%) and 14b/c (10.0%) being the two predominant strains. Forty patients who presented with recurrent episodes of syphilis had T. pallidum DNA identified from the initial and subsequent episodes, with five cases showing strain discrepancies. One patient had two strains identified from different clinical specimens collected in the same episode. Our findings show that 14f/f is the most common T. pallidum strain in Taiwan, where the prevalence of T. pallidum strains that show A2058G or A2059G mutation remains low. Different genotypes of T. pallidum can be identified in patients with recurrent episodes of syphilis.


Changes in health-related quality of life with long-term eltrombopag treatment in adults with persistent/chronic immune thrombocytopenia: Findings from the EXTEND study.
Patients with persistent/chronic immune thrombocytopenia (cITP) have low platelet counts, increased risk of bleeding and bruising, and often suffer from reduced health-related quality of life (HRQoL). cITP treatments may either improve HRQoL by increasing platelet counts or decrease it because of side effects. The open-label EXTEND study (June 2006 to July 2015) evaluated long-term safety, tolerability, and efficacy of eltrombopag (an oral thrombopoietin-receptor-agonist) in adults with cITP who completed a previous eltrombopag ITP trial. The final results of EXTEND were published and used to assess changes in patient-reported HRQoL over time and association between HRQoL and platelet response. Four validated HRQoL instruments were administered: SF-36v2 including physical component summary (PCS) and Mental Component Summary; Motivation and Energy Inventory Short Form (MEI-SF); Fatigue Subscale of FACIT (FACIT-Fatigue); and FACT-Thrombocytopenia Subscale Six-Item Extract (FACT-Th6). For the 302 patients enrolled, median duration of eltrombopag treatment was 2.37 years. All 4 HRQoL instruments demonstrated positive mean changes from baseline over time adjusted for patient baseline characteristics and rescue therapy use, and had positive association with platelet response (platelet count ≥30 × 109 /L; ≥50 × 109 /L; and ≥50 × 109 /L and >2 times baseline). Improvements from baseline started within 3 months and persisted through 5 years of treatment for FACIT-Fatigue and FACT-Th6 (P <.05 for nearly all time points); through 2.5 years for SF-36v2 PCS and less consistently for the MEI-SF. In conclusion, in addition to eltrombopag increasing platelet counts and reducing bleeding/bruising, it also alleviated fatigue, concerns about bleeding and bruising, and improved physical function in many patients, especially responders.


Fall prevention in the community: what older people say they need.
Uptake of and adherence to fall prevention interventions is often poor and we know little about how older people's perceptions of and beliefs about fall prevention interventions affect uptake. This study aimed to explore older people's perceptions of the facilitators and barriers to participation in fall prevention interventions. We undertook a qualitative study with older people who had taken part in, declined to participate or adhere to fall prevention interventions using semi-structured interviews (n=65), and 17 focus groups (n=122) with older people (including 32 South Asian and 30 Chinese older people) in primary and community care settings in the South of England. A number of factors acted as either barriers or facilitators to uptake of interventions. Older people also made recommendations for improving access to interventions. Community nurses are ideally placed to screen older people, identify those at risk of falling and refer them to appropriate interventions as well as providing health promotion and education.


Starvation-Induced Stress Response Is Critically Impacted by Ceramide Levels in Caenorhabditis elegans.
Our understanding of the cellular mechanisms by which animals regulate their response to starvation is limited, despite the strong relevance of the problem to major human health issues. The L1 diapause of Caenorhabditis elegans, where first-stage larvae arrest in response to a food-less environment, is an excellent system to study this mechanism. We found, through genetic manipulation and lipid analysis, that biosynthesis of ceramide, particularly those with longer fatty acid side chains, critically impacts animal survival during L1 diapause. Genetic interaction analysis suggests that ceramide may act in both insulin-IGF-1 signaling (IIS)-dependent and IIS-independent pathways to affect starvation survival. Genetic and expression analyses indicate that ceramide is required for maintaining the proper expression of previously characterized starvation-responsive genes, genes that are regulated by the IIS pathway and tumor suppressor Rb, and genes responsive to pathogen. These findings provide an important insight into the roles of sphingolipid metabolism, not only in starvation response, but also in aging and food-response-related human health problems.


Experimental investigation of the origin of fynbos plant community structure after fire.
Species in plant communities segregate along fine-scale hydrological gradients. Although this phenomenon is not unique to fynbos, this community regenerates after fire and therefore provides an opportunity to study the ecological genesis of hydrological niche segregation. Following wildfires at two field sites where we had previously mapped the vegetation and monitored the hydrology, seeds were moved experimentally in >2500 intact soil cores up and down soil-moisture gradients to test the hypothesis that hydrological niche segregation is established during the seedling phase of the life cycle. Seedling numbers and growth were then monitored and they were identified using DNA bar-coding, the first use of this technology for an experiment of this kind. At the site where niche segregation among Restionaceae had previously been found, the size of seedlings was significantly greater, the wetter the location into which they were moved, regardless of the soil moisture status of their location of origin, or of the species. Seedling weight was also significantly greater in a competition treatment where the roots of other species were excluded. No such effects were detected at the control site where niche segregation among Restionaceae was previously found to be absent. The finding that seedling growth on hydrological gradients in the field is affected by soil moisture status and by root competition shows that hydrological niche segregation could potentially originate in the seedling stage. The methodology, applied at a larger scale and followed-through for a longer period, could be used to determine whether species are differently affected by soil moisture.


Daratumumab, Carfilzomib, Lenalidomide, and Dexamethasone With Minimal Residual Disease Response-Adapted Therapy in Newly Diagnosed Multiple Myeloma.
The MASTER trial combined daratumumab, carfilzomib, lenalidomide, and dexamethasone (Dara-KRd) in newly diagnosed multiple myeloma (NDMM), using minimal residual disease (MRD) by next-generation sequencing (NGS) to inform the use and duration of Dara-KRd post-autologous hematopoietic cell transplantation (AHCT) and treatment cessation in patients with two consecutive MRD-negative assessments. This multicenter, single-arm, phase II trial enrolled patients with NDMM with planed enrichment for high-risk cytogenetic abnormalities (HRCAs). Patients received Dara-KRd induction, AHCT, and Dara-KRd consolidation, according to MRD status. MRD was evaluated by NGS at the end of induction, post-AHCT, and every four cycles (maximum of eight cycles) of consolidation. Primary end point was achievement of MRD negativity (< 10-5). Patients with two consecutive MRD-negative assessments entered treatment-free MRD surveillance. Among 123 participants, 43% had none, 37% had 1, and 20% had 2+ HRCA. Median age was 60 years (range, 36-79 years), and 96% had MRD trackable by NGS. Median follow-up was 25.1 months. Overall, 80% of patients reached MRD negativity (78%, 82%, and 79% for patients with 0, 1, and 2+ HRCA, respectively), 66% reached MRD < 10-6, and 71% reached two consecutive MRD-negative assessments during therapy, entering treatment-free surveillance. Two-year progression-free survival was 87% (91%, 97%, and 58% for patients with 0, 1, and 2+ HRCA, respectively). Cumulative incidence of MRD resurgence or progression 12 months after cessation of therapy was 4%, 0%, and 27% for patients with 0, 1, or 2+ HRCA, respectively. Most common serious adverse events were pneumonia (6%) and venous thromboembolism (3%). Dara-KRd, AHCT, and MRD response-adapted consolidation leads to high rate of MRD negativity in NDMM. For patients with 0 or 1 HRCA, this strategy creates the opportunity of MRD surveillance as an alternative to indefinite maintenance.


Comparison of calcaneal ultrasound and DXA to assess the risk of corticosteroid-induced osteoporosis: a cross-sectional study.
Patients on long-term oral corticosteroids have an increased risk of low bone mass and fragility fractures. Fracture risk rises soon after commencement of corticosteroid therapy and it is possible that these agents adversely influence bone architecture disproportionately to their effect on bone mass. The best means of assessing bone status in patients using corticosteroids remains uncertain, but quantitative ultrasound of the calcaneus may provide evidence of microarchitectural changes not detected by dual-energy X-ray absorptiometry (DXA). Patients with Crohn's disease have an increased risk of low bone mineral density (BMD), the etiology of which is multifactorial but includes corticosteroid use. We studied 118 consecutive patients with Crohn's disease, 21 of whom used continuous oral corticosteroids, 70 of whom were intermittent users, and 27 who had never used the drug. All patients received DXA of the lumbar spine, hip and calcaneus and quantitative ultrasound (QUS) of the calcaneus. The different techniques were compared using a femoral neck T-score < or = -1.5 as the threshold of corticosteroid-induced osteoporosis. When compared with the femoral neck T-score, there were no significant differences between the predictive values of lumbar spine DXA, calcaneal DXA or calcaneal QUS to identify low femoral neck BMD. However, the absolute T-score required to give similar discriminatory capacity to femoral neck T-score varied substantially (T= -0.81 to -1.5) between the different measurement techniques and sites.


Biomarkers for kidney transplant rejection.
The immune management of organ transplant recipients is imperfect. Beyond general dosing guidelines for immunosuppressive agents and clinical diagnostic tests for rejection or infection, there are few objective tools to determine the aggregate status of a patient's alloimmune response or protective immune capacity. The lack of prognostic precision significantly contributes to patient morbidity and reduces long-term allograft survival after kidney transplantation. Noninvasive biomarkers that could serve as predictive tools or surrogate end points for rejection might help clinicians individualize immunosuppression and allow for early intervention, ideally prior to clinically evident organ dysfunction. Although the growing understanding of organ rejection has provided numerous candidate biomarkers, none has been confirmed in robust validation studies as sufficiently useful to guide clinical practice independent of traditional clinical methods. In this Review, the general characteristics of biomarkers and surrogate end points; current biomarkers under active clinical investigation; and the prominent barriers to the translation of biomarkers into clinical practice are discussed.


A systematic review of the survival and complication rates of implant-supported fixed dental prostheses (FDPs) after a mean observation period of at least 5 years.
The objective of this systematic review was to assess the 5- and 10-year survival of implant-supported fixed dental prostheses (FDPs) and to describe the incidence of biological and technical complications. An electronic Medline search complemented by manual searching was conducted to identify prospective and retrospective cohort studies and case series on FDPs with a mean follow-up time of at least 5 years. Patients had to have been examined clinically at the follow-up visit. Failure and complication rates were analyzed using standard and random-effects Poisson regression models to obtain summary estimates of 5-year and 10-year survival and complication rates. The updated search provided 979 titles and 257 abstracts. Full-text analysis was performed for 90 articles resulting in a total 32 studies that met the inclusion criteria. Meta-analysis of these studies indicated an estimated survival of implants supporting FDPs of 95.6% after 5 years and 93.1% after 10 years. When machined surface implants were excluded from the analysis and only rough surface implants included, the survival rate increased to 97.2% after 5 years. The survival rate of implant-supported FDPs was 95.4% after 5 years and 80.1% after 10 years of function. When the analysis was done exclusively for metal-ceramic FDPs, hence the old gold-acrylic FDPs were excluded, the survival rate increased significantly. The survival rate of metal-ceramic implant-supported FDPs was 96.4% after 5 years and 93.9% after 10 years. Only 66.4% of the patients were free of any complications after 5 years. The most frequent complications over the 5-year observation period were fractures of the veneering material (13.5%), peri-implantitis and soft tissue complications (8.5%), loss of access hole restoration (5.4%), abutment or screw loosening (5.3%), and loss of retention of cemented FDPs (4.7%). It may be concluded that implant-supported fixed dental prostheses (FDPs) are a safe and predictable treatment method with high survival rates. However, biological and technical complications were frequent (33.6%). To minimize the incidence of complications, dental professionals should make great effort in choosing reliable components and materials for implant-supported FDPs and the patients should be placed in well-structured maintenance system after treatment.


Hippocampal long-term potentiation is supported by presynaptic and postsynaptic tyrosine receptor kinase B-mediated phospholipase Cgamma signaling.
Neurotrophins have been shown to play a critical role in activity-dependent synaptic plasticity such as long-term potentiation (LTP) in the hippocampus. Although the role of brain-derived neurotrophic factor (BDNF) and its tyrosine kinase receptor [tyrosine receptor kinase B (TrkB)] is well documented, it still remains unresolved whether presynaptic or postsynaptic activation of TrkB is involved in the induction of LTP. To address this question, we locally and specifically interfered with a downstream target of the TrkB receptor, phospholipase Cgamma (PLCgamma). We prevented PLCgamma signaling by overexpression of the PLCgamma pleckstrin homology (PH) domain with a Sindbis virus vector. The isolated PH domain has an inhibitory effect and thereby blocks endogenous PLCgamma signaling and consequently also IP3 production. Surprisingly, concurrent presynaptic and postsynaptic blockade of PLCgamma signaling was required to reduce LTP to levels comparable with those in TrkB and BDNF knock-out mice. Blockade of presynaptic or postsynaptic signaling alone did not result in a significant reduction of LTP.


HLA type-independent method to monitor polyoma BK virus-specific CD4 and CD8 T-cell immunity.
(Re)activation of quiescent viral diseases is a major problem in immunosuppressed transplant patients. Polyoma BK virus-associated nephropathy (PVAN) caused by active polyoma BK virus (BKV) infection became a main reason for graft loss in kidney transplantation. After diagnosis, most transplant centers react by reducing immunosuppression (IS) to allow the immune system to control the infection. However, the impact of reduced IS on BKV immunity is not well researched. Here we present an HLA type-independent method to monitor BKV-specific T-cell immunity. Applying our method, viral protein 1-specific CD4+ and CD8+ T-cell responses were detected in patients with serum BKV-DNA levels >250 000 copies/mL. In addition, specific T-cell responses were also found in allograft-infiltrating cells. The method can be used to assess the impact of decreased immunosuppression on BKV immunity and to clarify the role of specific T cells in the pathogenesis of PVAN. We strongly recommend its implementation in future clinical studies.


Validity of the Ages and Stages Questionnaire to detect risk of Developmental Coordination Disorder in preschoolers.
A reliable and valid screening tool for detecting children at risk for Developmental Coordination Disorder (DCD) is needed. The purpose of this study was to evaluate the ability of the Ages and Stages Questionnaire-Third Edition (ASQ-3) motor scales to detect children at risk for DCD in a community-based sample of children aged 3.5-5.5 years. One hundred and sixty parent-child pairs were recruited from community-based organizations. Children were eligible if they spoke English and had no known physical impairments. Eligible parents were asked to fill out the ASQ-3, following which their child's fine motor and gross motor proficiency was assessed using the Movement Assessment Battery for Children-Second Edition (MABC-2). DCD risk was defined as those children scoring at or below the 16th percentile on the MABC-2. Sensitivity and specificity of the fine and gross motor areas of the ASQ-3 were examined and referenced against the DCD risk classification. The ASQ-3 total motor score correlated moderately with overall standard score on the MABC-2 (r = 0.41; p < 0.001). Regardless of the ASQ-3 cut-off used, sensitivities of the ASQ-3 fine motor or gross motor scale to detect DCD risk were low at 21-47%, whereas specificities were high at 89-96%. Early identification of motor skill delays is important in order to intervene and hopefully prevent the associated negative health consequences. However, because of the low sensitivity of the ASQ-3 motor scales, these results suggest that the ASQ-3 is not an appropriate screening tool to identify children at risk for DCD in the preschool population.


Lack of concordance between residual viremia and viral variants driving de novo infection of CD4(+) T cells on ART.
In most patients, current antiretroviral therapy (ART) regimens can rapidly reduce plasma viral load. However, even after years of effective treatment, a significant proportion of patients show residual plasma viremia below the clinical detection limit. Although residual viremia might be associated with increased chronic immune activation and morbidity, its origin and its potential role in the replenishment of the viral reservoir during suppressive ART is not completely understood. We performed an in-depth genetic analysis of the total and episomal cell-associated viral DNA (vDNA) repertoire in purified CD4(+) T cell subsets of three HIV-infected individuals, and used phylogenetic analysis to explore its relationship with plasma viruses. The predominant proviral reservoir was established in naïve or memory (central and transitional) CD4(+) T cell subsets in patients harboring X4- or R5-tropic viruses, respectively. Regardless of the viral tropism, most plasma viruses detected under suppressive ART resembled the proviral reservoir identified in effector and transitional memory CD4(+) T-cell subsets in blood, suggesting that residual viremia originates from these cells in either blood or lymphoid tissue. Most importantly, sequences in episomal vDNA in CD4(+) T-cells were not well represented in residual viremia. Viral tropism determines the differential distribution of viral reservoir among CD4(+) T-cell subsets. In spite of viral tropism, the effector and transitional memory CD4(+) T-cells subsets are the main source of residual viremia during suppressive ART, even though their contribution to the total proviral pool is small. However, the lack of concordance between residual viremia and viral variants driving de novo infection of CD4(+) T cells on ART may reflect the predominance of defective plasma HIV RNA genomes. These findings highlight the need for monitoring the multiple viral RNA/DNA persistence markers, based on their differential contribution to viral persistence.


Infant mortality, season of birth and the health of older Puerto Rican adults.
The increasing prevalence of heart disease and diabetes among aging populations in low and middle income countries leads to questions regarding the degree to which endogenous early life exposures (exposures in utero) are important determinants of these health conditions. We devised a test using infant mortality (IMR) to verify if season of birth is a good indicator of early life (in utero) conditions that precipitate adult onset of disease. We linked annual IMR at the municipality (municipio) level from the late 1920s to early 1940s with individual birth year and place using a representative sample of older Puerto Rican adults (n = 1447) from the Puerto Rican Elderly: Health Conditions (PREHCO) study. We estimated the effects of season of birth on adult heart disease and diabetes for all respondents and then for respondents according to whether they were born when IMR was lower or higher, controlling for age, gender, obesity, respondent's educational level, adult behavior (smoking and exercise) and other early life exposures (childhood health, knee height and childhood socioeconomic status (SES)). The pattern of effects suggests that season of birth reflects endogenous causes: (1) odds of heart disease and diabetes were strong and significant for those born during the lean season in years when IMR was lower; (2) effects remained consistent even after controlling for other childhood conditions and adult behavior; but (3) no seasonality effects on adult health for adults born when IMR was higher. We conclude that in this population of older Puerto Rican adults there is continued support that the timing of adverse endogenous (in utero) conditions such as poor nutrition and infectious diseases is associated with adult heart disease and diabetes. It will be important to test the validity of these findings in other similar populations in the developing world.