Physical diagnoses in nursing home residents - is dementia or severity of dementia of importance?
Dementia and physical morbidity are primary reasons for nursing home admission globally. However, data on physical morbidity in nursing home residents with and without dementia are scarce. The first aim of the present study was to explore whether presence and severity of dementia were related to the number of physical diagnoses in nursing home residents. The second aim was to explore if the severity of dementia was associated with having registered the most frequent complexes of physical diagnoses when controlling for physical health and demographic factors. A total of 2983 Norwegian nursing home residents from two cross-sectional samples from 2004/2005 and 2010/2011 were included in the analysis. By the use of assessment scales, the severity of dementia (Clinical Dementia Rating), physical health (General Medical Health Rating), activities of daily living (Physical Self-Maintenance Scale) and neuropsychiatric symptoms (Neuropsychiatric Inventory Nursing Home) were determined. Physical diagnoses and medications were assembled from the medical records. The physical diagnoses were categorized into complexes, using the ICD-10 chapters. Linear mixed models and generalized linear mixed models were estimated. Residents with dementia were registered with fewer physical diagnoses than residents without dementia. The frequency of physical diagnoses decreased with increasing severity of dementia. Cardiovascular, musculoskeletal and endocrine, nutritional and metabolic diagnoses were the most common complexes of physical diagnoses in individuals with and without dementia. The odds of having cardiovascular and musculoskeletal diagnoses increased for males and decreased for females with increasing severity of dementia, in contrast to endocrine diagnoses where the odds increased for both genders. Increasing severity of dementia in nursing home residents may complicate the diagnostics of physical disease. This might reflect a need for more attention to the registration of physical diagnoses in nursing home residents with dementia.


Optimization of a technology-supported physical activity intervention for breast cancer survivors: Fit2Thrive study protocol.
Fit2Thrive is a theory-guided physical activity promotion trial using the Multiphase Optimization Strategy (MOST) to test efficacy for improving physical activity of five technology-supported physical activity promotion intervention components among breast cancer survivors. This trial will recruit 256 inactive breast cancer survivors nationwide. All participants will receive the core intervention which includes a Fitbit and standard self-monitoring Fit2Thrive smartphone application which will be downloaded to their personal phone. Women will be randomized to one of 32 conditions in a factorial design involving five factors with two levels: support calls (No vs. Yes), app type (standard vs. deluxe), text messaging (No vs. Yes), online gym (No vs. Yes) and Fitbit Buddy (No vs. Yes). The proposed trial examines the effects of the components on physical activity at 12 and 24weeks. Results will support the selection of a final package of intervention components that has been optimized to maximize physical activity and is subject to an upper limit of cost. The optimized intervention will be tested in a future trial. Fit2Thrive is the first trial to use the MOST framework to develop and test a physical activity promotion intervention in breast cancer survivors and will lead to an improved understanding of how to effectively change survivors' physical activity. These findings could result in more scalable, effective physical activity interventions for breast cancer survivors, and, ultimately, improve health and disease outcomes.


Comparative study of the biological properties of Trypanosoma cruzi I genotypes in a murine experimental model.
Chagas disease is an endemic zoonosis in Latin America and caused by the parasite Trypanosoma cruzi. This kinetoplastid displays remarkable genetic variability, allowing its classification into six Discrete Typing Units (DTUs) from TcI to TcVI. T. cruzi I presents the broadest geographical distribution in the continent and has been associated to severe forms of cardiomyopathies. Recently, a particular genotype associated to human infections has been reported and named as TcIDOM (previously named TcIa-b). This genotype shows to be clonal and adapted to the domestic cycle but so far no studies have determined the biological properties of domestic (TcIDOM) and sylvatic TcI strains (previously named TcIc-e). Hence, the aim of this study was to untangle the biological features of these genotypes in murine models. We infected ICR-CD1 mice with five TcI strains (two domestic, two sylvatic and one natural mixture) and determined the course of infection during 91 days (acute and chronic phase of the disease) in terms of parasitemia, tissue tropism, immune response (IgG titers) and tissue invasion by means of histopathology studies. Statistically significant differences were observed in terms of parasitemia curves and prepatent period between domestic (TcIDOM) and sylvatic strains. There were no differences in terms of IgG antibodies response across the mice infected with the five strains. Regarding the histopathology, our results indicate that domestic strains present higher parasitemias and low levels of histopathological damage. In contrast, sylvatic strains showed lower parasitemias and high levels of histopathological damage. These results highlight the sympatric and behavioral differences of domestic and sylvatic TcI strains; the clinical and epidemiological implications are herein discussed.


Impaired HLA capping capacity of peripheral blood lymphocytes in Duchenne muscular dystrophy.
The cap capacity in nine Duchenne muscular dystrophy (DMD) patients and in 23 healthy male subjects (14 adults and nine neonates) has been investigated by inducing capping of HLA molecules. The evaluation of capping percentages ranged in healthy controls from 44 to 61 with a mean value of 53.39 +/- 4.89, while DMD patients displayed cap capacity of percentages varying from 15 to 38 with a mean value of 31.5 +/- 7.42. Statistical significance of the differences between the two groups, assessed by the Mann-Whitney U test, was p less than 0.00003. A correlation was found in DMD patients between cap capacity and age (tau = +0.657, p = 0.012). The results confirm previous reports of Ig capping impairment noted in B cells of the whole lymphocyte population, supporting the hypothesis of a systemic cellular defect in DMD patients. The data obtained suggest that HLA capping could overcome some of the technical difficulties of Ig capping and could be used as a diagnostic aid in antenatal detection of DMD.


Respiratory syncytial virus influences NF-kappaB-dependent gene expression through a novel pathway involving MAP3K14/NIK expression and nuclear complex formation with NF-kappaB2.
A member of the Paramyxoviridae family of RNA viruses, respiratory syncytial virus (RSV), is a leading cause of epidemic respiratory tract infection in children. In children, RSV primarily replicates in the airway mucosa, a process that alters epithelial cell chemokine expression, thereby inducing airway inflammation. We investigated the role of the mitogen-activated protein kinase kinase kinase 14/NF-kappaB-inducing kinase (NIK) in the activation of NF-kappaB-dependent genes in alveolus-like A549 cells. RSV infection induces a time dependent increase of NIK mRNA and protein expression that peaks 12 to 24 h after viral exposure. Immunoprecipitation kinase assays indicate that NIK kinase activity is activated even more rapidly (within 6 h of RSV adsorption) associated with an endogenous approximately 50-kDa NF-kappaB2 substrate. Because NIK associates with IKKalpha to mediate processing of the 100-kDa NF-kappaB2 precursor into its 52-kDa DNA binding isoform ("p52"), the effects of RSV on NIK complex formation with IKKalpha and NF-kappaB2 were determined by coimmunoprecipitation assay. We find that NIK, IKKalpha, and both 100 kDa- and 52-kDa NF-kappaB2 isoforms strongly complex 15 h after exposure to RSV at times subsequent to NIK kinase activation. Western immunoblot and microaffinity DNA pull-down assays showed a parallel increase in nuclear translocation and DNA binding of the NF-kappaB2-Rel B complex. Interestingly, we make the novel observations that NIK also transiently translocates into the nucleus complexed with 52-kDa NF-kappaB2. Small interfering RNA-mediated NIK "knock-down" blocked RSV-inducible 52-kDa NF-kappaB2 processing and interfered with the early activation of a subset of NF-kappaB-dependent genes, indicating the importance of this activation pathway in the genomic NF-kappaB response to RSV. Together, these data indicate that RSV infection rapidly activates the noncanonical NF-kappaB activation pathway prior to the more potent canonical pathway activation. This appears to be through a novel mechanism involving induction of NIK kinase activity, expression, and nuclear translocation of a ternary complex with IKKalpha and processed NF-kappaB2.


Increases in Condomless Sex in the Swiss HIV Cohort Study.
Condomless sex is a key driver of sexually transmitted diseases. In this study, we assess the long-term changes (2000-2013) of the occurrence of condomless sex among human immunodeficiency virus (HIV)-infected individuals enrolled in the Swiss HIV Cohort study. The frequencies with which HIV-infected individuals reported condomless sex were either stable or only weakly increasing for 2000-2008. For 2008-2013, these rates increased significantly for stable relationships among heterosexuals and men who have sex with men (MSM) and for occasional relationships among MSM. Our results highlight the increasing public health challenge posed by condomless sex and show that condomless sex has been increasing even in the most recent years.


Assessing the value of pediatric hospitalist programs: the perspective of hospital leaders.
There has been a rapid increase in the number of pediatric hospitalist programs in the United States. As most pediatric hospitalist services are believed to be subsidized by hospitals, gaining a better understanding of the rationale for these subsidies is critical to the future success and existence of these programs. Our objective was to determine the rationale for pediatric hospitalist program subsidies from the perspective of hospital leaders. A survey was mailed to hospital executives from a national sample of 112 hospitals between October 2007 and February 2008. The overall response rate was 69% (N = 77). Twelve hospitals no longer used hospitalists to provide care for children, leaving 65 hospitals for the analysis. The majority of hospital leaders indicated they subsidize their pediatric hospitalist program (78%, n = 51) and the average proportion subsidized was 49% of program costs. The majority of hospitals (82%, n = 40) do not plan to phase out the subsidy of hospitalists over time, as they do not anticipate their program will be able to cover its costs. Hospital leaders provided a broad rational for this subsidization but most commonly cited the nonmonetary benefits of patient (83%, n = 39) and referring physician satisfaction (81%, n = 38) as reason for their investment. Despite the fact that most pediatric hospitalist programs are unable to cover their costs, the majority of hospitals plan to continue subsidizing these programs. Discussions of the value added by hospitalists should not presume that hospital investment in hospitalist programs is based on monetary benefits alone.


Exposure assessment to ochratoxin A in Catalonia (Spain) based on the consumption of cereals, nuts, coffee, wine, and beer.
Ochratoxin A (OTA) was analysed in composite samples of cereal-based baby foods, beer, breakfast cereals (corn- and rice and wheat-based), loaf bread, peanuts and pistachios. Foodstuffs were collected in hypermarkets and supermarkets from 12 cities in the Spanish region of Catalonia, and composite samples were prepared for analysis involving liquid-liquid extraction, followed by immunoaffinity column clean-up and HPLC with fluorescence detection. Consumption data for the selected foodstuffs were collected by means of a food-frequency questionnaire. The studied population was grouped by age in infants, children, adolescents and adults; and exposure to OTA through the specified foodstuffs, and through wine and coffee, was assessed. Exposure assessment was done through deterministic and probabilistic modelling of the contamination and consumption data. OTA occurrence and mean of positive samples (ng g(-1) or ng ml(-1), for beer) were the following: 8.7% and 0.233 in baby foods; 88.7% and 0.022 in beer; 2.8% and 0.728 in corn-based breakfast cereals; 25% and 0.293 in wheat-based breakfast cereals; 12.9% and 0.283 in loaf bread; 41.7% and 0.241 in peanuts; and 2.9% and 0.228 in pistachios. The median estimated daily intake of OTA through the foodstuffs by each age group were below the latest provisional tolerable daily intakes (PTDIs) of 17 and 14 ng kg(-1) bw day(-1) recommended by the European Food Safety Authority (EFSA) in 2006 and the Joint FAO/WHO Expert Committee on Food Additives (JECFA) in 2007, respectively, ranging from 1% and 2% of those values in adolescents and children, to 3% and 11% in adults and infants.


Utility of bile duct brushings for the early detection of cholangiocarcinoma in patients with primary sclerosing cholangitis.
In patients with primary sclerosing cholangitis, there are no reliable markers or imaging modalities to detect malignant changes or early cholangiocarcinoma when curative interventions may still be possible. Clinical features and outcomes were evaluated for 47 patients who underwent 101 endoscopic retrograde cholangiopancreatography (ERCP) procedures with bile duct brushings and cytopathologic examination for the detection of malignant changes between January 2001 and 2004. Bile duct cytology was characterized as unsatisfactory, benign, atypical, or malignant. Fifteen patients had at least one sample that was abnormal. Patients with abnormal findings were older (P = 0.03); otherwise the groups were similar. Three of six patients with marked atypia have undergone transplantation for these abnormal findings, and of these, two had cholangiocarcinoma and one had no cancer detected in the explant. Of the nine patients with focal atypia, seven are doing well, one underwent transplantation for marked atypia seen on brushings at another hospital, and the other underwent transplantation for decompensated cirrhosis. Two patients with benign findings developed cholangiocarcinoma, whereas 29 others have not. Cytopathologic examination of bile duct brushings taken at ERCP may be useful for the early detection of malignant changes in patients with primary sclerosing cholangitis. However, long-term prospective studies are needed to confirm the utility of surveillance ERCP and brushings.


Automated characterization of perceptual quality of clinical chest radiographs: validation and calibration to observer preference.
The authors previously proposed an image-based technique [Y. Lin et al. Med. Phys. 39, 7019-7031 (2012)] to assess the perceptual quality of clinical chest radiographs. In this study, an observer study was designed and conducted to validate the output of the program against rankings by expert radiologists and to establish the ranges of the output values that reflect the acceptable image appearance so the program output can be used for image quality optimization and tracking. Using an IRB-approved protocol, 2500 clinical chest radiographs (PA/AP) were collected from our clinical operation. The images were processed through our perceptual quality assessment program to measure their appearance in terms of ten metrics of perceptual image quality: lung gray level, lung detail, lung noise, rib-lung contrast, rib sharpness, mediastinum detail, mediastinum noise, mediastinum alignment, subdiaphragm-lung contrast, and subdiaphragm area. From the results, for each targeted appearance attribute/metric, 18 images were selected such that the images presented a relatively constant appearance with respect to all metrics except the targeted one. The images were then incorporated into a graphical user interface, which displayed them into three panels of six in a random order. Using a DICOM calibrated diagnostic display workstation and under low ambient lighting conditions, each of five participating attending chest radiologists was tasked to spatially order the images based only on the targeted appearance attribute regardless of the other qualities. Once ordered, the observer also indicated the range of image appearances that he/she considered clinically acceptable. The observer data were analyzed in terms of the correlations between the observer and algorithmic rankings and interobserver variability. An observer-averaged acceptable image appearance was also statistically derived for each quality attribute based on the collected individual acceptable ranges. The observer study indicated that, for each image quality attribute, the averaged observer ranking strongly correlated with the algorithmic ranking (linear correlation coefficient R > 0.92), with highest correlation (R = 1) for lung gray level and the lowest (R = 0.92) for mediastinum noise. There was a strong concordance between the observers in terms of their rankings (i.e., Kendall's tau agreement > 0.84). The observers also generally indicated similar tolerance and preference levels in terms of acceptable ranges, as 85% of the values were close to the overall tolerance or preference levels and the differences were smaller than 0.15. The observer study indicates that the previously proposed technique provides a robust reflection of the perceptual image quality in clinical images. The results established the range of algorithmic outputs for each metric that can be used to quantitatively assess and qualify the appearance quality of clinical chest radiographs.


The Bcl-2/Bcl-XL inhibitor ABT-737 promotes death of retinoblastoma cancer cells.
Retinoblastoma is a malignant tumor that usually develops in early childhood. During retinoblastoma spreading, RB1 gene inactivation is followed by additional genomic modifications which progressively lead to resistance of tumor cells to death. Drugs that act at downstream levels of death signaling pathways should therefore be interesting in killing retinoblastoma cells. ABT-737, a BH3 mimetic molecule effective at the mitochondrial level, has been shown to induce apoptosis in different human tumoral cell lines as well as in primary patient-derived cells, and in a mouse xenograph model. In this report, we analyzed the pro-death effect of ABT-737 on two human retinoblastoma cell lines, Y79 and WERI-Rb, as well as on the mouse photoreceptor cell line 661W. We observed that ABT-737 was very effective as a single agent in inducing human WERI-Rb cells apoptosis without affecting the mouse 661W photoreceptor cells. However human Y79 cells were resistant to ABT-737, as a probable consequence of the absence of Bax. The high sensitivity of WERI-Rb to ABT-737 can be increased by downregulating Mcl-1 using the proteasome inhibitor MG-132. Preliminary analysis in primary mouse retinoblastoma tumoral cell lines predicts high sensitivity to ABT-737. Our data suggest that ABT-737 or related compounds could be a highly effective drug in the treatment of some retinoblastomas.


Growth hormone stimulates tyrosine phosphorylation of insulin receptor substrate-1.
Growth hormone (GH) produces insulin-like effects in rat adipocytes that have been deprived of GH for at least 3 h. The effect of a saturating concentration of GH is qualitatively and quantitatively similar to that produced by 2-4 ng/ml insulin but differs from that of insulin in the respect that adipocytes become refractory to prolonged or repeated stimulation with GH. Since activation of tyrosine kinase is an early event in the action of both hormones, we investigated the possibility that GH stimulation of tyrosine phosphorylation of some protein in the insulin transduction cascade might result in the similar effect of the two hormones. Adipocytes were preincubated for 3 h in the absence of hormones and then reincubated without or with 500 ng/ml GH or 4-400 ng/ml insulin for 10 min. The cells were lysed with an equal volume of buffer containing 1% SDS and preheated to 100 degrees C. Proteins were separated by electrophoresis on 7.5% polyacrylamide gels and transferred to nitrocellulose membranes, and tyrosine-phosphorylated proteins were detected using anti-phosphotyrosine antiserum coupled to horseradish peroxidase and reagents to produce chemiluminescence. The faint band seen at 185 kDa in control lanes was increased by GH treatment in five independent experiments. Insulin produced a similar effect at a concentration of 4 ng/ml, and phosphorylation increased in a dose-related manner in cells treated with higher concentrations of insulin. A prominent approximately 95-kDa band that is probably not the beta subunit of the insulin receptor was also seen in GH-treated cells. The beta subunit of the insulin receptor has similar electrophoretic mobility to the 95-kDa protein, but was not phosphorylated to an extent that allowed detection when insulin was added at concentrations below 400 ng/ml. Phosphorylation of the 185- and 95-kDa bands was evident within 1 min after addition of GH, persisted for at least 30 min, and was equally prominent in sensitive and refractory cells. Antiserum to IRS-1 immunoprecipitated the tyrosine-phosphorylated 185-kDa protein. The data suggest that IRS-1 is a substrate for a GH-activated tyrosine kinase, possibly JAK-2, which may account for the insulin-like effects of GH. The data further suggest that refractoriness to insulin-like stimulation by GH may result from an additional GH-dependent action that is distinct from phosphorylation of IRS-1.


Coverage of atypical antipsychotics among medicare drug plans in the state of washington: changes between 2007 and 2008.
To examine changes in the cost and coverage of atypical antipsychotics among Medicare prescription drug plans and Medicare advantage plans in the state of Washington. Coverage and cost data were obtained in February 2007 and 2008 from the Medicare Prescription Drug Plan Finder, an online database administered by the Centers for Medicare and Medicaid Services. Premiums, deductibles, out-of-pocket costs, and coverage limits were compared for prescription drug plans (PDPs) and for Medicare advantage plans (MAPs). The number of PDPs in the state of Washington fell slightly from 57 in 2007 to 53 in 2008, while the number of MAPs rose from 43 in 2007 to 52 in 2008. In 2008, the mean monthly drug premium increased by 15% among PDPs and by 20% among MAPs. Mean copayments for the majority of atypical antipsychotics increased from 2007 to 2008. More plans added quantity limits for atypical antipsychotics, but use of other pharmacy management tools varied by type of plan and antipsychotic. PDP and MAP participants in the state of Washington paid more for atypical antipsychotics in 2008 than they did in 2007. Affordability of atypical antipsychotics continues to be a concern, particularly for beneficiaries who are not eligible for Medicaid or the low-income subsidy.


Effects of referral to a specialist for headache.
Patients consulting neurological outpatient clinics for headaches that were found not to be due to a serious structural lesion were followed up one year afterwards. Considerable improvement in symptoms was found in the sample. This was only partly attributable to any medical treatment received at the clinics or subsequently from a general practitioner. Improvement was associated with previously expressed satisfaction with the clinic consultation, and a nonspecific `placebo' response is postulated.


Helical distortion in tryptophan- and lysine-anchored membrane-spanning alpha-helices as a function of hydrophobic mismatch: a solid-state deuterium NMR investigation using the geometric analysis of labeled alanines method.
We used solid-state deuterium NMR spectroscopy and geometric analysis of labeled alanines to investigate the structure and orientation of a designed synthetic hydrophobic, membrane-spanning alpha-helical peptide that is anchored within phosphatidylcholine (PC) bilayers using both Trp and Lys side chains near the membrane/water interface. The 23-amino-acid peptide consists of an alternating Leu/Ala core sequence that is expected to be alpha-helical, flanked by aromatic and then cationic anchors at both ends of the peptide: acetyl-GKALW(LA)(6)LWLAKA-amide (KWALP23). The geometric analysis of labeled alanines method was elaborated to permit the incorporation and assignment of multiple alanine labels within a single synthetic peptide. Peptides were incorporated into oriented bilayers of dilauroyl- (di-C12:0-), dimyristoyl- (di-C14:0-), or dioleoyl- (di-C18:1c-) PC. In the C12:0 and C14:0 lipids, the (2)H-NMR quadrupolar splittings for the set of six core alanines could not be fit to a canonical undistorted alpha-helix. Rather, we found that a model containing a helical distortion, such as a localized discontinuity or "kink" near the peptide and bilayer center, could fit the data for KWALP23 in these shorter lipids. The suggestion of helix distortion was confirmed by (2)H-NMR spectra for KWALP23 in which Leu(8) was changed to deuterated Ala(8). Further analysis involving reexamination of earlier data led to a similar conclusion that acetyl-GWW(LA)(8)LWWA-amide (WALP23) is distorted in dilauroyl-PC, allowing significant improvement in the fitting of the (2)H-NMR results. In contrast, WALP23 and KWALP23 are well represented as undistorted alpha-helices in dioleoyl-PC, suggesting that the distortion could be a response to hydrophobic mismatch between peptide and lipids.


Adjacent-segment effects of lumbar cortical screw-rod fixation versus pedicle screw-rod fixation with and without interbody support.
Cortical screw-rod (CSR) fixation has emerged as an alternative to the traditional pedicle screw-rod (PSR) fixation for posterior lumbar fixation. Previous studies have concluded that CSR provides the same stability in cadaveric specimens as PSR and is comparable in clinical outcomes. However, recent clinical studies reported a lower incidence of radiographic and symptomatic adjacent-segment degeneration with CSR. No biomechanical study to date has focused on how the adjacent-segment mobility of these two constructs compares. This study aimed to investigate adjacent-segment mobility of CSR and PSR fixation, with and without interbody support (lateral lumbar interbody fusion [LLIF] or transforaminal lumbar interbody fusion [TLIF]). A retroactive analysis was done using normalized range of motion (ROM) data at levels adjacent to single-level (L3-4) bilateral screw-rod fixation using pedicle or cortical screws, with and without LLIF or TLIF. Intact and instrumented specimens (n = 28, all L2-5) were tested using pure moment loads (7.5 Nm) in flexion, extension, lateral bending, and axial rotation. Adjacent-segment ROM data were normalized to intact ROM data. Statistical comparisons of adjacent-segment normalized ROM between two of the groups (PSR followed by PSR+TLIF [n = 7] and CSR followed by CSR+TLIF [n = 7]) were performed using 2-way ANOVA with replication. Statistical comparisons among four of the groups (PSR+TLIF [n = 7], PSR+LLIF [n = 7], CSR+TLIF [n = 7], and CSR+LLIF [n = 7]) were made using 2-way ANOVA without replication. Statistical significance was set at p < 0.05. Proximal adjacent-segment normalized ROM was significantly larger with PSR than CSR during flexion-extension regardless of TLIF (p = 0.02), or with either TLIF or LLIF (p = 0.04). During lateral bending with TLIF, the distal adjacent-segment normalized ROM was significantly larger with PSR than CSR (p < 0.001). Moreover, regardless of the types of screw-rod fixations (CSR or PSR), TLIF had a significantly larger normalized ROM than LLIF in all directions at both proximal and distal adjacent segments (p ≤ 0.04). The use of PSR versus CSR during single-level lumbar fusion can significantly affect mobility at the adjacent segment, regardless of the presence of TLIF or with either TLIF or LLIF. Moreover, the type of interbody support also had a significant effect on adjacent-segment mobility.


Amniotic fluid is not useful for diagnosis of congenital Trypanosoma cruzi infection.
Although Trypanosoma cruzi can be transmitted transplacentally and induce congenital infection, no data are available about the presence of this parasite in human amniotic fluid. We examined 8, 19, and 4 amniotic fluid samples (collected at delivery or by aspiration of gastric content of neonates) from control uninfected mothers (M-B-), infected mothers delivering uninfected newborns (M+B-), and mothers of confirmed congenital cases (M+B+), respectively. Polymerase chain reaction (PCR), using nuclear and kinetoplastic DNA primers (Tcz1-Tcz2 and 121-122), were negative for all control M-B- samples, but positive for 5 of 19 M+B- and 2 of 4 M+B+ samples. To determine the number of parasites in the positive samples, real-time PCR using S35/S36 kinetoplastic DNA was performed. Only one M+B+ sample presented a high parasitic DNA amount, whereas the other six PCR-positive samples displayed traces of T. cruzi DNA. In conclusion, the release of parasites in amniotic fluid is probably a rare event that cannot be helpful for the routine diagnosis of congenital Chagas disease.


A combined in vivo neurochemical and electrophysiological analysis of the effect of high-frequency stimulation of the subthalamic nucleus on 5-HT transmission.
Movement disability in advanced Parkinson's disease (PD) can be treated by high frequency stimulation (HFS) of the subthalamic nucleus (STN) but some patients experience psychiatric side-effects including depression, which is strongly linked to decreases in 5-hydroxytryptamine (5-HT). The current study investigated the effect of bilateral STN HFS on extracellular 5-HT in brain regions of anesthetized and freely moving rats as measured with microdialysis. Parallel in vivo electrophysiological experiments allowed a correlation of changes in extracellular 5-HT with the firing of 5-HT neurons. Bilateral STN HFS decreased (by up to 25%) extracellular levels of 5-HT in both striatum and medial prefrontal cortex of anesthetized rats. STN HFS also decreased extracellular 5-HT in the medial prefrontal cortex of freely moving rats. This decrease in extracellular 5-HT persisted after turning off the stimulation, and was present in dopamine-denervated rats. As with changes in extracellular 5-HT, in anesthetized rats STN HFS evoked a decrease in the in vivo firing of midbrain raphe 5-HT neurons that also persisted after cessation of stimulation. These data provide neurochemical evidence for an inhibition of 5-HT neurotransmission by STN HFS, which may contribute to its psychiatric side effects and guide therapeutic options.


The effect of neuromuscular block and noxious stimulation on hypnosis monitoring during sevoflurane anesthesia.
There are conflicting results on the influence of neuromuscular block (NMB) on the bispectral index (BIS). We investigated the influence of two degrees of NMB on BIS, Alaris auditory-evoked potential index (AAI), and the electromyogram (EMG) obtained with needle electrodes from the frontal and temporal muscles, immediately adjacent to the BIS-sensor. Twenty patients were anesthetized with sevoflurane, titrated for 30 min to an end-tidal concentration of 1.2% (baseline). Rocuronium was infused to 50% (partial) and 95% (profound) depression of the first twitch in a train-of-four response, the order being randomly chosen. Noxious tetanic electrical stimulation was applied at four occasions: 1) at baseline (control measurement), 2 and 3) at each degree of NMB, and 4) after neostigmine reversal. BIS, AAI, and EMG were obtained 2 min before and 2 min after each noxious stimulation. Median BIS and AAI at baseline were 44 (39-50) and 15 (14-16), respectively. The two degrees of NMB did not affect BIS, AAI, and EMG before noxious stimulation. In contrast, profound NMB altered the BIS and AAI responses to noxious stimulation when compared with partial NMB, (BIS P = 0.01, AAI P < 0.01), after neostigmine reversal (BIS P < 0.01, AAI P = 0.01) and compared with baseline (BIS P = 0.08, AAI P = 0.02). No significant increase in EMG was found. BIS and AAI responses to noxious tetanic electrical stimulation are affected by the degree of NMB during sevoflurane anesthesia whereas NMB does not affect BIS or AAI in the absence of noxious stimulation.


Ovarian cancer risk associated with inherited inflammation-related variants.
The importance of inflammation pathways to the development of many human cancers prompted us to examine the associations between single-nucleotide polymorphisms (SNP) in inflammation-related genes and risk of ovarian cancer. In a multisite case-control study, we genotyped SNPs in a large panel of inflammatory genes in 930 epithelial ovarian cancer cases and 1,037 controls using a custom array and analyzed by logistic regression. SNPs with P < 0.10 were evaluated among 3,143 cases and 2,102 controls from the Follow-up of Ovarian Cancer Genetic Association and Interaction Studies (FOCI) post-GWAS collaboration. Combined analysis revealed association with SNPs rs17561 and rs4848300 in the interleukin gene IL1A which varied by histologic subtype (P(heterogeneity) = 0.03). For example, IL1A rs17561, which correlates with numerous inflammatory phenotypes, was associated with decreased risk of clear cell, mucinous, and endometrioid subtype, but not with the most common serous subtype. Genotype at rs1864414 in the arachidonate 5-lipoxygenase ALOX5 was also associated with decreased risk. Thus, inherited variation in IL1A and ALOX5 seems to affect ovarian cancer risk which, for IL1A, is limited to rarer subtypes. Given the importance of inflammation in tumorigenesis and growing evidence of subtype-specific features in ovarian cancer, functional investigations will be important to help clarify the importance of inherited variation related to inflammation in ovarian carcinogenesis.


What design and material factors impact the wear and corrosion performance in total elbow arthroplasties?
The survivorship of total elbow arthroplasties is lower than surgeons and patients would like it to be, especially in patients with posttraumatic arthritis of the elbow. To improve durability, it is important to understand the failure modes of existing implants. Total elbow arthroplasties were designed primarily for low-demand rheumatoid patients. As surgical indications have extended to more active patient populations, the mechanical performance of current designs must meet an increased mechanical burden. Evaluating the degree to which they do this will guide conclusions about which contemporary devices might still meet the need and, as importantly, what design and material changes might be needed to improve performance. WHERE ARE WE NOW?: The reasons for failures of total elbow arthroplasties include infection, loosening, polyethylene wear, locking mechanism failure, periprosthetic fracture, implant fracture, and instability. Implant design factors that have influenced wear include implant constraint, material, coatings, and metal backing. Surgical factors associated with increased wear and subsequent total elbow arthroplasty failure include soft tissue balancing and restoration of alignment and implant positioning. WHERE DO WE NEED TO GO?: A clear need exists for improving the performance of total elbow arthroplasty. Many of the failures that have limited the survivorship of elbow arthroplasties thus far are mechanical in nature with wear-related problems a dominating influence. Much of what we know about the results of total elbow arthroplasty is from small studies frequently involving the designer of the implant. The establishment of total elbow arthroplasty registries coupled with the increasing regulatory burden of postmarket surveillance would lead to a better understanding of the complications and survivorship of elbow arthroplasties. Another primary goal must be to achieve a better understanding of the biomechanics of the normal elbow and how the mechanics are altered after the insertion of elbow arthroplasty components. HOW DO WE GET THERE?: Improving the performance and survivorship of total elbow arthroplasty will require the integration of clinical and implant performance data gained through the establishment of registries with a concerted basic science effort to better understand the functional loads across the joint and to incorporate these loads into experimental and computational models to allow assessment of design and material changes intended to improve durability.


The management of behavioural and psychological symptoms of dementia in residential homes: does Tai Chi have any role for people with dementia?
Dementia is a common illness that is increasing in frequency and set to challenge the resources and expertise of health and social care services over the coming years. Increasingly, there has been interest in the management of behavioural and psychological symptoms of dementia (BPSD), as they are both common and associated with a range of negative outcomes. BPSD are associated with the admission of people with dementia to care homes. Limited resources and lack of knowledge in permanent care settings often lead to BPSD being managed with antipsychotic medications, which are associated with significant morbidity and mortality. There is evidence for the benefits of exercise within care home settings, although only a few studies include those with cognitive impairment. Tai Chi is a mind-body exercise combining relaxed physical movement and meditation, and has been suggested to have many health benefits. This article discusses the rationale and available options for treating BPSD and the current practice and reviews the literature regarding the benefits of exercise and, in particular, Tai Chi in the management of BPSD.


ST proteins, a new family of plant tandem repeat proteins with a DUF2775 domain mainly found in Fabaceae and Asteraceae.
Many proteins with tandem repeats in their sequence have been described and classified according to the length of the repeats: I) Repeats of short oligopeptides (from 2 to 20 amino acids), including structural cell wall proteins and arabinogalactan proteins. II) Repeats that range in length from 20 to 40 residues, including proteins with a well-established three-dimensional structure often involved in mediating protein-protein interactions. (III) Longer repeats in the order of 100 amino acids that constitute structurally and functionally independent units. Here we analyse ShooT specific (ST) proteins, a family of proteins with tandem repeats of unknown function that were first found in Leguminosae, and their possible similarities to other proteins with tandem repeats. ST protein sequences were only found in dicotyledonous plants, limited to several plant families, mainly the Fabaceae and the Asteraceae. ST mRNAs accumulate mainly in the roots and under biotic interactions. Most ST proteins have one or several Domain(s) of Unknown Function 2775 (DUF2775). All deduced ST proteins have a signal peptide, indicating that these proteins enter the secretory pathway, and the mature proteins have tandem repeat oligopeptides that share a hexapeptide (E/D)FEPRP followed by 4 partially conserved amino acids, which could determine a putative N-glycosylation signal, and a fully conserved tyrosine. In a phylogenetic tree, the sequences clade according to taxonomic group. A possible involvement in symbiosis and abiotic stress as well as in plant cell elongation is suggested, although different STs could play different roles in plant development. We describe a new family of proteins called ST whose presence is limited to the plant kingdom, specifically to a few families of dicotyledonous plants. They present 20 to 40 amino acid tandem repeat sequences with different characteristics (signal peptide, DUF2775 domain, conservative repeat regions) from the described group of 20 to 40 amino acid tandem repeat proteins and also from known cell wall proteins with repeat sequences. Several putative roles in plant physiology can be inferred from the characteristics found.


Transporter gene acquisition and innovation in the evolution of Microsporidia intracellular parasites.
The acquisition of genes by horizontal transfer can impart entirely new biological functions and provide an important route to major evolutionary innovation. Here we have used ancient gene reconstruction and functional assays to investigate the impact of a single horizontally transferred nucleotide transporter into the common ancestor of the Microsporidia, a major radiation of intracellular parasites of animals and humans. We show that this transporter provided early microsporidians with the ability to steal host ATP and to become energy parasites. Gene duplication enabled the diversification of nucleotide transporter function to transport new substrates, including GTP and NAD+, and to evolve the proton-energized net import of nucleotides for nucleic acid biosynthesis, growth and replication. These innovations have allowed the loss of pathways for mitochondrial and cytosolic energy generation and nucleotide biosynthesis that are otherwise essential for free-living eukaryotes, resulting in the highly unusual and reduced cells and genomes of contemporary Microsporidia.


Proactive interception and care of Frailty and Multimorbidity in older persons: the experience of the European Innovation Partnership on Active and Healthy Ageing and the response of Parma Local Health Trust and Lab through European Projects.
According to the 2018 European Union Ageing Report, the demographic profile of the European population is projected to be older. Aging cannot be considered a homogeneous process, and only in certain cases is "successful", with maintained functional ability, which is determined by intrinsic capacity, the environment, and their interaction. When intrinsic capacity is lost, elders with chronic diseases develop frailty, a condition with high-risk of disability. Old-age dependency-ratio is projected to increase from 29.6% to 51.2% in the EU in 2070: thus, the need of new approaches targeting the prevention of disability. Numerous studies are conducted in the European Innovation Partnership on Active and Healthy Ageing and addressing identification, treatment, coordination and integration of care in frail older subjects. SUNFRAIL is aimed at developing a model, good practices and tools to improve the identification, prevention and care of frailty and management of multimorbidity. SPRINTT is testing the effectiveness of a multi-component treatment able to treat frailty and sarcopenia. VIGOUR, a project aimed at strengthening integrated-care in different contexts of European Countries, verifies enablers and obstacles encountered in the real world by these good practices. Through the creation of Parma-Lab and Frailty-Team in the Academic-Hospital of Parma combined with the contribution of Parma Health-Trust in the "Community Health-Centers", the Projects were translated into Health Services Arena. This response bridging European Studies and clinical practice, aims to early detecting and caring 75-year older citizens with frailty and multimorbidity, living in the community, not institutionalized and at risk of hospitalization and mobility ADL-disability.


Plasticity of calcium channels in dendritic spines.
Voltage-sensitive Ca2+ channels (VSCCs) constitute a major source of calcium ions in dendritic spines, but their function is unknown. Here we show that R-type VSCCs in spines of rat CA1 pyramidal neurons are depressed for at least 30 min after brief trains of back-propagating action potentials. Populations of channels in single spines are depressed stochastically and synchronously, independent of channels in the parent dendrite and other spines, implying that depression is the result of signaling restricted to individual spines. Induction of VSCC depression blocks theta-burst-induced long-term potentiation (LTP), indicating that postsynaptic action potentials can modulate synaptic plasticity by tuning VSCCs. Induction of depression requires [Ca2+] elevations and activation of L-type VSCCs, which activate Ca2+/calmodulin-dependent kinase II (CaMKII) and a cyclic adenosine monophosphate (cAMP)-dependent pathway. Given that L-type VSCCs do not contribute measurably to Ca2+ influx in spines, they must activate downstream effectors either directly through voltage-dependent conformational changes or via [Ca2+] microdomains.


Cardiovascular health effects of oral and pulmonary exposure to multi-walled carbon nanotubes in ApoE-deficient mice.
Exposure to high aspect ratio nanomaterials, such as multi-walled carbon nanotubes (MWCNTs) may be associated with increased risk of atherosclerosis, pulmonary disease, and cancer. In the present study, we investigated the cardiovascular and pulmonary health effects of 10 weeks of repeated oral or pulmonary exposures to MWCNTs (4 or 40μg each week) in Apolipoprotein E-deficient (ApoE-/-) mice fed a Western-type diet. Intratracheal instillation of MWCNTs was associated with oxidative damage to DNA in lung tissue and elevated levels of lipid peroxidation products in plasma, whereas the exposure only caused a modest pulmonary inflammation in terms of increased numbers of lymphocytes in bronchoalveolar lavage fluid. Ultrasound imaging in live animals revealed an increase in the inner and outer wall thickness of the aortic arch at 10 weeks after pulmonary exposure to MWCNTs, which may suggest artery remodelling. However, we did not find accelerated plaque progression in the aorta or the brachiocephalic artery by histopathology. Furthermore, repeated oral exposure to MWCNTs did not cause changes in the composition of gut microbiota of exposed mice. Collectively, this study indicates that repeated pulmonary exposure to MWCNTs was associated with oxidative stress, whereas cardiovascular effects encompassed remodelling of the aorta wall.


Attitudes of internal medicine subspecialty fellows toward primary care.
Subspecialists deliver a substantial proportion of primary care but little is known about how their training affects their attitudes toward this role. We surveyed a department of medicine to determine fellows' (N = 34) attitudes toward primary care and how these compared with the attitudes of house staff (N = 45) and faculty (N = 66). Continuous, coordinated, and accessible care as departmental policy was almost unanimously endorsed by all physicians. In contrast, fellows less often supported the provision of such care for their own patients in actual clinical situations. Fellows were also less likely than either house staff or faculty to endorse primary care attributes for their own patients. Departments of medicine should examine how negative attitudes toward primary care develop in subspecialty fellows and whether these attitudes persist after fellowship.


Effect of carbohydrate ingestion on exercise performance and carbohydrate metabolism in persons with spinal cord injury.
Carbohydrate ingestion during exercise and as a pre-exercise bolus improves exercise performance in able-bodied athletes. Little is known about the potential for carbohydrate ingestion to improve exercise performance in athletes with spinal cord injury (SCI), nor the potential physiological limitations of such a practice resulting from an SCI. Therefore, this study investigated the effects of carbohydrate ingestion on exercise performance in physically active and athletic persons with SCI. Six participants with complete SCI (neurological level of lesion ranging from C(6) to T(7)) and normal glucose tolerance were studied twice during 60 min of arm cranking at 65% of peak oxygen consumption followed by a 20-min time trial with the ingestion of either a carbohydrate drink (CHO trial: 0.5 g CHO kg(-1) body weight in 500 ml) or placebo (PLA trial) applied in a double-blind counter-balanced manner. The participants with tetraplegia had sufficient neurological function to permit voluntary arm-cranking exercise. There was no difference in time-trial performance between CHO and PLA trials (P > 0.05). The results suggest that carbohydrate ingestion in persons with SCI does not improve exercise performance.


Effect of ischemic preconditioning on lactate accumulation and running performance.
Repeated bouts of ischemia followed by reperfusion (i.e., ischemic preconditioning (IPC)) protect against damage after a myocardial infarction. Recent observational data indicate that IPC improves exercise performance. However, no previous study has examined potential underlying mechanisms for this effect of IPC. Therefore, we examined the potential of IPC to improve 5-km running time trial performance and reduce lactate accumulation during an incremental exercise test. In a randomized, crossover study, 13 healthy men performed running exercise, which was preceded by IPC (4 × 5-min 220 mm Hg bilateral leg occlusion) or a control intervention (C) (4 × 5-min 20 mm Hg bilateral leg occlusion). Participants performed a graded maximal treadmill running test, starting with five 3-min submaximal stages (10-14 km·h), followed by increments of 1 km·h every 2 min to 16 km·h, followed by an incline of the treadmill of 2% every 2 min. Blood lactate was examined at each 3-min stage. After 45 min of rest in the supine position, subjects then performed a 5-km running time trial. We found similar submaximal gas parameters during running exercise with both interventions. The overall increase in blood lactate during the submaximal stages was 1.07 ± 0.11 mmol·L lower when exercise was preceded with IPC versus C (P = 0.023). The 5-km running time trial was completed in a time that was 34 s faster after IPC versus C (95% confidence interval, 5-64 s; P = 0.027). IPC improves 5-km time trial performance in healthy male individuals. Moreover, we found that IPC is associated with an attenuated rise in blood lactate concentration at submaximal level during an incremental running test. This could indicate that IPC allows for higher work rates and thus improves time trial performance.


State healthcare reform: an arena for the osteopathic medical profession's influence.
While healthcare reform proposals are debated at the national level, states continue to propose and implement reform measures to address Medicaid, health insurance, universal coverage and access, medical liability, and cost-containment. The authors examine the shared responsibility of the federal and state governments for healthcare regulation and the surprising number of powers that reside with the states. They review the major barriers to state reform represented by restrictions within Medicaid and the Employee Retirement Income Security Act of 1974 (ERISA) legislation. Established state programs in Maryland, Hawaii, and Arizona are revisited, and innovative reforms in Oregon, Tennessee, and Washington are examined. Finally, the authors concentrate on the reform measures under way in the five most heavily DO-populated states, pointing out the potential for one of the big three (Michigan, Pennsylvania, and Ohio) to emerge as a model for the larger states. They urge osteopathic physicians to exert influence, based on their record of serving the Medicaid and other underserved populations, in state settings where they can be most effective.


Child sexual abuse, links to later sexual exploitation/high-risk sexual behavior, and prevention/treatment programs.
This paper reviews the literature on the nature and incidence of child sexual abuse, explores the link between child sexual abuse and later sexual exploitation, and reviews the literature on prevention strategies and effective interventions in child sexual abuse services. Our understanding of the international epidemiology of child sexual abuse is considerably greater than it was just 10 years ago, and studies from around the world are examined. Childhood sexual abuse can involve a wide number of psychological sequelae, including low self-esteem, anxiety, and depression. Numerous studies have noted that child sexual abuse victims are vulnerable to later sexual revictimization, as well as the link between child sexual abuse and later engagement in high-risk sexual behaviour. Survivors of child sexual abuse are more likely to have multiple sex partners, become pregnant as teenagers, and experience sexual assault as adults. Various models which attempt to account for this inter-relationship are presented; most invoke mediating variables such as low self-esteem, drug/alcohol use, PTSD and distorted sexual development. Prevention strategies for child sexual abuse are examined including media campaigns, school-based prevention programmes, and therapy with abusers. The results of a number of meta-analyses are examined. However, researchers have identified significant methodological limitations in the extant research literature that impede the making of recommendations for implementing existing therapeutic programmes unreservedly.


Parent-teacher agreement on ADHD symptoms across development.
Parent-teacher agreement on attention-deficit/hyperactivity disorder (ADHD) symptom ratings ranges from low to moderate. Most studies evaluating parent-teacher agreement have not assessed measurement invariance across raters. Hence, it is unclear whether discordance across raters is due to differing ADHD constructs across raters or other factors (e.g., subjective differences across raters). Additionally, the effect of development on parent-teacher agreement is relatively unknown. To address these limitations, the present study used parent and teacher ADHD ratings from a large (N = 6,659) developmentally diverse (ages 4-17) sample. Using exploratory structural equation modeling on half the sample, and then confirmatory factor analysis (CFA) on the other half of the sample, confirmed a 2-factor structure with significant cross-loadings for the 18 ADHD symptoms. CFA invariance analyses demonstrated that the 2-factor symptom structure was similar across raters and age groups. After confirming measurement invariance, the correlation between latent factors within and across raters was examined for each age group as well as across age groups. Parents reported greater severity of ADHD symptoms than did teachers, and both parents and teachers reported higher levels of hyperactivity/impulsivity in younger children than in older children and consistent levels of inattention across development. Finally, correlations between parent-teacher ratings of like factors were weak for inattention and moderate-strong for hyperactivity/impulsivity, and the magnitude of parent-teacher agreement did not vary across development. In conclusion, while parent and teacher ratings of ADHD behaviors are only weakly to moderately correlated, each reporter provides unique and valid clinical information as it relates to ADHD symptom presentation.


Genome-wide association study of lung function decline in adults with and without asthma.
Genome-wide association studies have identified determinants of chronic obstructive pulmonary disease, asthma, and lung function level; however, none have addressed decline in lung function. We conducted the first genome-wide association study on the age-related decrease in FEV(1) and its ratio to forced vital capacity (FVC) stratified a priori by asthma status. Discovery cohorts included adults of European ancestry (1,441 asthmatic and 2,677 nonasthmatic participants: the Epidemiological Study on the Genetics and Environment of Asthma, the Swiss Cohort Study on Air Pollution and Lung and Heart Disease in Adults, and the European Community Respiratory Health Survey). The associations of FEV(1) and FEV(1)/FVC ratio decrease with 2.5 million single nucleotide polymorphisms (SNPs) were estimated. Thirty loci were followed up by in silico replication (1,160 asthmatic and 10,858 nonasthmatic participants: Atherosclerosis Risk in Communities, the Framingham Heart Study, the British 1958 Birth Cohort, and the Dutch Asthma Study). Main signals identified differed between asthmatic and nonasthmatic participants. None of the SNPs reached genome-wide significance. The association between the height-related gene DLEU7 and FEV(1) decrease suggested for nonasthmatic participants in the discovery phase was replicated (discovery, P = 4.8 × 10(-6); replication, P = .03), and additional sensitivity analyses point to a relation to growth. The top ranking signal, TUSC3, which is associated with FEV(1)/FVC ratio decrease in asthmatic participants (P = 5.3 × 10(-8)), did not replicate. SNPs previously associated with cross-sectional lung function were not prominently associated with decline. Genetic heterogeneity of lung function might be extensive. Our results suggest that genetic determinants of longitudinal and cross-sectional lung function differ and vary by asthma status.


The combined effects of Trigonella and Allium extracts in the regulation of hyperthyroidism in rats.
The combined effects of Trigonella foenum-graecum and Allium sativum extracts were evaluated for their ameliorative potential in the L-thyroxine-induced hyperthyroidic rat model to contribute to an understanding of interaction between the two extracts. The investigation was carried out using two different doses. A comparison was made with the response of individual plant extracts at the previously studied effective dose in adult Wistar rats rendered hyperthyroidic by daily injections of L-thyroxine (300 microg/kg body wt., s.c.). Propylthiouracil (PTU), an antithyroid drug, was used as a reference compound. Alterations in serum triiodothyronine (T3), thyroxine (T4), glucose, hepatic glucose-6-phosphatase (G-6-Pase) and oxygen consumption were studied as end parameters. Superoxide dismutase (SOD), catalase (CAT) activities, lipid peroxidation (LPO) and reduced glutathione (GSH) were examined to reveal any toxic effects of the drugs. The combined effects of Trigonella and Allium at 200 and 500 mg/kg body wt. respectively, were equipotent as compared to the individual extracts in lowering the serum concentrations of T3 and T4 in hyperthyroidic rats. Our findings reveal that some plant extracts in combination may not always prove to be synergistic. It is therefore suggested that Trigonella foenum-graecum and Allium sativum extracts may be used individually and not together in the regulation of hyperthyroidism.


A statistical approach for intensity loss compensation of confocal microscopy images.
In this paper, a probabilistic technique for compensation of intensity loss in confocal microscopy images is presented. For single-colour-labelled specimen, confocal microscopy images are modelled as a mixture of two Gaussian probability distribution functions, one representing the background and another corresponding to the foreground. Images are segmented into foreground and background by applying Expectation Maximization algorithm to the mixture. Final intensity compensation is carried out by scaling and shifting the original intensities with the help of parameters estimated for the foreground. Since foreground is separated to calculate the compensation parameters, the method is effective even when image structure changes from frame to frame. As intensity decay function is not used, complexity associated with estimation of the intensity decay function parameters is eliminated. In addition, images can be compensated out of order, as only information from the reference image is required for the compensation of any image. These properties make our method an ideal tool for intensity compensation of confocal microscopy images that suffer intensity loss due to absorption/scattering of light as well as photobleaching and the image can change structure from optical/temporal section-to-section due to changes in the depth of specimen or due to a live specimen. The proposed method was tested with a number of confocal microscopy image stacks and results are presented to demonstrate the effectiveness of the method.


Self-organizing actin patterns shape cytoskeletal cortex organization.
Living systems rely, for biological function, on the spatiotemporal organization of their structures. Cellular order naturally emerges by dissipation of energy. Consequently, energy-consuming processes operating far from thermodynamic equilibrium are a necessary condition to enable biological systems to respond to environmental cues that allow their transitions between different steady-states. Such self-organization was predicted for the actin cytoskeleton in theoretical considerations and has repeatedly been observed in cell-free systems. We now demonstrate in our recent work how self-organizing actin patterns such as vortices, stars, and asters may allow cells to adjust their membrane architecture without affecting their cell mechanical properties.


Exercise capacity and mortality in hypertensive men with and without additional risk factors.
We assessed the association between exercise capacity and mortality in hypertensive men with and without additional cardiovascular risk factors. A cohort of 4631 hypertensive veterans, who successfully completed a graded exercise test at the Veterans Affairs Medical Center in Washington, DC, and Palo Alto, California, was followed for 7.7+/-5.4 years (35,629 person-years) for all-cause mortality. Fitness categories were established based on peak metabolic equivalent (MET) levels achieved. In each fitness category, we defined individuals with and without additional cardiovascular risk factors. Exercise capacity was the strongest predictor of all-cause mortality. The adjusted mortality risk was 13% lower for every 1-MET increase in exercise capacity. Compared with the very low fit (< or =5.0 MET), the adjusted risk was 34% lower for those achieving 5.1 to 7.0 MET (low fit; hazard ratio: 0.66; CI: 0.58 to 0.76; P<0.001), 59% lower for the moderate fit (7.1 to 10.0 MET; hazard ratio: 0.41; CI: 0.35 to 0.50; P<0.001), and 71% lower for the high-fit category (>10.0 MET; hazard ratio: 0.29; CI: 0.21 to 0.40; P<0.001). Within the very-low-fit category, mortality risk was 47% higher for those with additional risk factors compared with individuals with no risk factors. This risk was eliminated for those in the next fitness category (5.1 to 7.0 MET) and was progressively reduced for the moderate and high-fit categories regardless of the presence or absence of additional risk factors. In conclusion, exercise capacity was the strongest predictor of all-cause mortality in hypertensive men. The increased risk imposed by low fitness and additional cardiovascular risk factors was eliminated by relatively small increases in exercise capacity and declined progressively with higher exercise capacity.


N-Substituted analogues of S-nitroso-N-acetyl-D,L-penicillamine: chemical stability and prolonged nitric oxide mediated vasodilatation in isolated rat femoral arteries.
Previous studies show that linking acetylated glucosamine to S-nitroso-N-acetyl-D,L-penicillamine (SNAP) stabilizes the molecule and causes it to elicit unusually prolonged vasodilator effects in endothelium-denuded, isolated rat femoral arteries. Here we studied the propanoyl (SNPP; 3 carbon side-chain), valeryl (SNVP; 5C) and heptanoyl (SNHP; 7C) N-substituted analogues of SNAP (2C), to further investigate other molecular characteristics that might influence chemical stability and duration of vascular action of S-nitrosothiols. Spectrophotometric analysis revealed that SNVP was the most stable analogue in solution. Decomposition of all four compounds was accelerated by Cu(II) and cysteine, and neocuproine, a specific Cu(I) chelator, slowed decomposition of SNHP. Generation of NO from the compounds was confirmed by electrochemical detection at 37 degrees C. Bolus injections of SNAP (10 microl; 10(-8)-10(-3) M) into the perfusate of precontracted, isolated rat femoral arteries taken from adult male Wistar rats (400-500 g), caused concentration-dependent, transient vasodilatations irrespective of endothelial integrity. Equivalent vasodilatations induced by SNVP and SNHP were transient in endothelium-intact vessels but failed to recover to pre-injection pressures at moderate and high concentrations (10(-6)-10(-3) M) in those denuded of endothelium. This sustained effect (> 1 h) was most prevalent with SNHP and was largely reversed by the NO scavenger, haemoglobin. We suggest that increased lipophilicity of SNAP analogues with longer sidechains facilitates their retention by endothelium-denuded vessels; subsequent slow decomposition within the tissue generates sufficient NO to cause prolonged vasodilatation. This is a potentially useful characteristic for targeting NO delivery to areas of endothelial damage.


Syphilis screening in the antenatal care: a cross-sectional study from Botswana.
Congenital syphilis is recognized as a substantial public health problem in Sub-Saharan Africa. The aim of this study was to determine the prevalence of syphilis among antenatal care attendees in Botswana and to contribute to knowledge about the challenges facing the syphilis-screening programme. In a cross-sectional study, 703 antenatal care attendees at 13 health facilities in Gaborone, Botswana were interviewed and examined. Venous blood samples were collected for the identification of syphilis infection. The antenatal records were used to obtain information on any screening, diagnosis and treatment of syphilis that had been done earlier in the current pregnancy. Active syphilis was found in 32 (5%) of the attendees. Among 546 women coming for a repeat antenatal care visit, 71 (13%) had not been screened for syphilis. Uptake late in pregnancy, delayed treatment and a high rate of seroconversion after testing were other identified obstacles to the effective prevention of congenital syphilis. Syphilis prevalence among pregnant women in Botswana remains high, and there is still much to be gained by improving the effectiveness of the syphilis screening and treatment programme. Earlier antenatal care attendance, rapid on-site testing, improved partner treatment and a repeat test late in pregnancy to manage incident cases are important goals for patients, health care workers and health authorities.


Innovative delivery systems for migraine: the clinical utility of a transdermal patch for the acute treatment of migraine.
Migraine is a disabling, painful primary headache disorder that is associated with various combinations of neurological, gastrointestinal, autonomic and pain symptoms. Gastrointestinal disturbances associated with migraine, including nausea and vomiting, affect a majority of migraineurs and often result in a delay in taking or avoidance of pharmacological intervention. Gastric stasis and vomiting may lead to delayed or inconsistent absorption of orally administered medications. Many migraineurs awake early in the morning with their attack progressing and already associated with nausea and vomiting. As a result, there is a need for a novel, non-invasive, non-oral delivery system for fast and effective acute treatment of migraine. There are two non-oral delivery systems currently available in the US for the acute treatment of migraine: three nasal sprays and two injectable formulations. Although nasal sprays depend partially on nasal mucosal absorption, a significant amount of drug is swallowed, transits the stomach and is absorbed in the small intestine, which is not as rapid or effective a route of delivery for those migraineurs with gastric stasis. Sumatriptan is rapidly absorbed by subcutaneous injection with or without a needle, but the invasiveness and discomfort of the delivery, the high incidence of adverse events and the high recurrence rate all limit its use for many patients. Iontophoretic delivery of medication is a non-invasive transdermal approach that uses small amounts of electrical current to promote rapid movement of the ionized drug through the skin and into the systemic circulation. This delivery bypasses hepatic first-pass metabolism and also avoids gastric transit delay and slowing of small intestinal absorption associated with gastrointestinal stasis in migraineurs. Two pharmacokinetic studies have demonstrated that iontophoretic transdermal delivery of sumatriptan results in rapid and consistent achievement of therapeutic plasma concentrations. These studies also suggest that, by avoiding patient exposure to a rapid rise in and high plasma concentrations of sumatriptan as seen with injectable sumatriptan, transdermal delivery using iontophoresis may significantly reduce typical triptan-related adverse events. A large, randomized, double-blind, placebo-controlled, multicentre clinical trial showed statistically significant efficacy, good tolerability and virtually no triptan-related adverse events. Iontophoretic delivery of sumatriptan, with a novel transdermal patch device, offers patients a migraine-specific medication that is non-invasive and non-oral. Clinically, transdermal delivery provides rapid and effective relief of migraine while bypassing the gastrointestinal tract, with minimal classic triptan-related adverse effects. This unique approach facilitates the rapid absorption of this migraine-specific triptan, which should improve the chances of consistently achieving a therapeutic plasma concentration of sumatriptan, resulting in effective migraine relief.


The Romberg ratio in people with multiple sclerosis.
Postural control relies on the integration of inputs from the visual, somatosensory and vestibular systems which are frequently impaired in people with Multiple Sclerosis (PwMS). In this situation, examining the Romberg ratio can be useful. This parameter can be interpreted as a gross indicator of a vestibular and proprioceptive contribution to postural control. Therefore, the primary objective of the current study was to examine whether the Romberg ratio differs between MS fallers, non-fallers and neurological disability levels. In addition, we clarified the association between the Romberg ratio values with validated gait and the balance tests in PwMS. Romberg ratio values were calculated according to the sway rate, total sway area and center of pressure (CoP) path length. The patient group included 542 PwMS (337 women) with a mean age of 42.3 (S.D=13.8). In terms of fall status, significant differences were observed between the faller (n=287) and non-faller (n=255) groups solely in terms of the Romberg ratio-ellipse sway area: 2.76 (S.D=2.46) vs. 2.24 (S.D=2.01), P-value=0.01. A significant increase in the Romberg ratio was found between the severe group (n=50), the very mild (n=245), mild (n=186) and moderate (n=61) groups for each of the three Romberg ratio quotients. Significant weak correlation scores were found between the Romberg ratio-ellipse sway area and all walking and balance outcome measures; the Pearson's rho ranged from 0.172 to 0.270. The present data suggest that an elevated Romberg ratio quotient, especially according to the sway area, is an indicator of poor walking and balance capabilities in PwMS.


In Vitro and In Vivo Biocompatibility Of ReOss® in Powder and Putty Configurations.
This study evaluated comparatively two configurations (powder and putty) of a composite biomaterial based on PLGA (Poly(lactide-co-glycolide)/nanoescale hydroxyapatite (ReOss®, Intra-Lock International) through microscopic morphology, in vitro cytotoxicity, biocompatibility and in vivo response as a bone substitute. SEM and EDS characterized the biomaterials before/after grafting. Cytocompatibility was assessed with murine pre-osteoblasts. Osteoconductivity and biocompatibility were evaluated in White New Zealand rabbits. Both configurations were implanted in the calvaria of eighteen animals in non-critical size defects, with blood clot as the control group. After 30, 60 and 90 days, the animals were euthanized and the fragments containing the biomaterials and controls were harvested. Bone blocks were embedded in paraffin (n=15) aiming at histological and histomorphometric analysis, and in resin (n=3) aiming at SEM and EDS. Before implantation, the putty configuration showed both a porous and a fibrous morphological phase. Powder revealed porous particles with variable granulometry. EDS showed calcium, carbon, and oxygen in putty configuration, while powder also showed phosphorus. After implantation EDS revealed calcium, carbon, and oxygen in both configurations. The materials were considered cytotoxic by the XTT test. Histological analysis showed new bone formation and no inflammatory reaction at implant sites. However, the histomorphometric analysis indicated that the amount of newly formed bone was not statistically different between experimental groups. Although both materials presented in vitro cytotoxicity, they were biocompatible and osteoconductive. The configuration of ReOss® affected morphological characteristics and the in vitro cytocompatibility but did not impact on the in vivo biological response, as measured by the present model.


Assessment of radiographic and clinical outcomes of an articulating expandable interbody cage in minimally invasive transforaminal lumbar interbody fusion for spondylolisthesis.
OBJECTIVE The inability to significantly improve sagittal parameters has been a limitation of minimally invasive surgery for transforaminal lumbar interbody fusion (MIS TLIF). Traditional cages have a limited capacity to restore lordosis. This study evaluates the use of a crescent-shaped articulating expandable cage (Altera) for MIS TLIF. METHODS This is a retrospective review of 1- and 2-level MIS TLIF. Radiographic outcomes included differences in segmental and lumbar lordosis, disc height, evidence of fusion, and any endplate violations. Clinical outcomes included the numeric rating scale for leg and back pain and the Oswestry Disability Index (ODI) for low-back pain. RESULTS Thirty-nine patients underwent single-level MIS TLIF, and 5 underwent 2-level MIS TLIF. The mean age was 63.1 years, with 64% women. On average, spondylolisthesis was corrected by 4.3 mm (preoperative = 6.69 mm, postoperative = 2.39 mm, p < 0.001), the segmental angle was improved by 4.94° (preoperative = 5.63°, postoperative = 10.58°, p < 0.001), and segmental height increased by 3.1 mm (preoperative = 5.09 mm, postoperative = 8.19 mm, p < 0.001). At 90 days after surgery the authors observed the following: a smaller postoperative sagittal vertical axis was associated with larger changes in back pain at 90 days (r = -0.558, p = 0.013); a larger decrease in spondylolisthesis was associated with greater improvements in ODI and back pain scores (r = -0.425, p = 0.043, and r = -0.43, p = 0.031, respectively); and a larger decrease in pelvic tilt (PT) was associated with greater improvements in back pain (r = -0.548, p = 0.043). For the 1-year PROs, the relationship between the change in PT and changes in ODI and numeric rating scale back pain were significant (r = 0.612, p = 0.009, and r = -0.803, p = 0.001, respectively) with larger decreases in PT associated with larger improvements in ODI and back pain. Overall for this study there was a 96% fusion rate. Fourteen patients were noted to have endplate violation on intraoperative fluoroscopy during placement of the cage. Only 3 of these had progression of their subsidence, with an overall subsidence rate of 6% (3 of 49) visible on postoperative CT. CONCLUSIONS The use of this expandable, articulating, lordotic, or hyperlordotic interbody cage for MIS TLIF provides a significant restoration of segmental height and segmental lordosis, with associated improvements in sagittal balance parameters. Patients treated with this technique had acceptable levels of fusion and significant reductions in pain and disability.


Calcium-Induced Differentiation of Human Colon Adenomas in Colonoid Culture: Calcium Alone versus Calcium with Additional Trace Elements.
Previous murine studies have demonstrated that dietary Aquamin, a calcium-rich, multi-mineral natural product, suppressed colon polyp formation and transition to invasive tumors more effectively than calcium alone when provided over the lifespan of the animals. In the current study, we compared calcium alone to Aquamin for modulation of growth and differentiation in human colon adenomas in colonoid culture. Colonoids established from normal colonic tissue were examined in parallel. Both calcium alone at 1.5 mmol/L and Aquamin (provided at 1.5 mmol/L calcium) fostered differentiation in the adenoma colonoid cultures as compared with control (calcium at 0.15 mmol/L). When Aquamin was provided at an amount delivering 0.15 mmol/L calcium, adenoma differentiation also occurred, but was not as complete. Characteristic of colonoids undergoing differentiation was a reduction in the number of small, highly proliferative buds and their replacement by fewer but larger buds with smoother surface. Proliferation marker (Ki67) expression was reduced and markers of differentiation (CK20 and occludin) were increased along with E-cadherin translocalization to the cell surface. Additional proteins associated with differentiation/growth control [including histone-1 family members, certain keratins, NF2 (merlin), olfactomedin-4 and metallothioneins] were altered as assessed by proteomics. Immunohistologic expression of NF2 was higher with Aquamin as compared with calcium at either concentration. These findings support the conclusions that (i) calcium (1.5 mmol/L) has the capacity to modulate growth and differentiation in large human colon adenomas and (ii) Aquamin delivering 0.15 mmol/L calcium has effects on proliferation and differentiation not observed when calcium is used alone at this concentration. Cancer Prev Res; 11(7); 413-28. ©2018 AACR.


Targeted therapy with antineoplastons A10 and AS2-1 of high-grade, recurrent, and progressive brainstem glioma.
Brainstem glioma carries the worst prognosis of all malignancies of the brain. Most patients with brainstem glioma fail standard radiation therapy and chemotherapy and do not survive longer than 2 years. Treatment is even more challenging when an inoperable tumor is of high-grade pathology (HBSG). The objective of this report is to summarize the outcome of patients with HBSG treated with antineoplastons in 4 phase 2 trials. The following group of 18 patients was evaluable: 4 patients with glioblastomas and 14 patients with anaplastic HBSG. Fourteen patients had diffuse intrinsic tumors. Twelve patients suffered from recurrence, and 6 patients did not have radiation therapy or chemotherapy. Antineoplastons, which consist of antineoplaston A10 (A10I) and AS2-1 injections, were given in escalating doses by intravenous injections. The median duration of antineoplaston administration was 5 months, and the average dosage of A10I was 9.22 g/kg/d and of AS2-1 was 0.31 g/kg/d. Responses were assessed by gadolinium-enhanced magnetic resonance imaging and positron emission tomography. The overall survival at 2 and 5 years was 39% and 22%, respectively, and maximum survival was more than 17 years for a patient with anaplastic astrocytoma and more than 5 years for a patient with glioblastoma. Progression-free survival at 6 months was 39%. Complete response was achieved in 11%, partial response in 11%, stable disease in 39%, and progressive disease in 39% of patients. Antineoplastons were tolerated very well with 1 case of grade 4 toxicity (reversible anemia). Antineoplastons contributed to more than a 5-year survival in recurrent diffuse intrinsic glioblastomas and anaplastic astrocytomas of the brainstem in a small group of patients.


Access to a syllabus of human hemoglobin variants (1996) via the World Wide Web.
Information on mutations in human hemoglobin is important in many efforts, including understanding the pathophysiology of hemoglobin diseases, developing therapies, elucidating the dynamics of sequence alterations inhuman populations, and dissecting the details of protein structure/function relationships. Currently, information is available on a large number of mutations and variants, but is distributed among thousands of papers. In an effort to organize this voluminous data set, two Syllabi have been prepared compiling succinct information on human hemoglobin abnormalities. In both of these, each entry provides amino acid and/or DNA sequence alterations, hematological and clinical data, methodology used for characterization, ethnic distribution, and functional properties and stability of the hemoglobin, together with appropriate literature references. A Syllabus of Human Hemoglobin Variants (1996) describes 693 abnormal hemoglobins resulting from alterations in the alpha-, beta-, gamma-, and delta-globin chains, including special abnormalities such as double mutations, hybrid chains, elongated chains, deletions, and insertions. We have converted this resource to an electronic form that is accessible via the World Wide Web at the Globin Gene Server (http://globin.cse.psu.edu). Hyperlinks are provided from each entry in the tables of variants to the corresponding full description. In addition, a simple query interface allows the user to find all entries containing a designated word or phrase. We are in the process of converting A Syllabus of Thalassemia Mutations (1997) to a similar electronic format.


Mental illness and suicide. A case-control study in east Taiwan.
As part of the Taiwan Aboriginal Study Project, a case-control study of suicide among two aboriginal groups and the Han Chinese was carried out in East Taiwan. Biographical reconstructive interviews were conducted for consecutive suicides from each of the three ethnic groups (a total of 116 suicides), 113 of whom were matched with two controls for age, sex, and area of residence. In all three groups, a high proportion of suicides suffered from mental illness before committing suicide (97% to 100%). The two most prevalent psychiatric disorders were depression and alcoholism, and the most common comorbid pattern was depression with substance use disorders. The risk for suicide was significantly associated with all of these psychiatric conditions, previous suicide attempts, and a family history of suicide and depression. Fifty-one percent of all suicides had consulted medical professionals in the previous month. Despite the widely different rates of depressive illness and alcoholism in different cultures previously reported, the psychiatric antecedents of suicide are the same in the West and the East.


Temporal clustering analysis of endothelial cell gene expression following exposure to a conventional radiotherapy dose fraction using Gaussian process clustering.
The vascular endothelium is considered as a key cell compartment for the response to ionizing radiation of normal tissues and tumors, and as a promising target to improve the differential effect of radiotherapy in the future. Following radiation exposure, the global endothelial cell response covers a wide range of gene, miRNA, protein and metabolite expression modifications. Changes occur at the transcriptional, translational and post-translational levels and impact cell phenotype as well as the microenvironment by the production and secretion of soluble factors such as reactive oxygen species, chemokines, cytokines and growth factors. These radiation-induced dynamic modifications of molecular networks may control the endothelial cell phenotype and govern recruitment of immune cells, stressing the importance of clearly understanding the mechanisms which underlie these temporal processes. A wide variety of time series data is commonly used in bioinformatics studies, including gene expression, protein concentrations and metabolomics data. The use of clustering of these data is still an unclear problem. Here, we introduce kernels between Gaussian processes modeling time series, and subsequently introduce a spectral clustering algorithm. We apply the methods to the study of human primary endothelial cells (HUVECs) exposed to a radiotherapy dose fraction (2 Gy). Time windows of differential expressions of 301 genes involved in key cellular processes such as angiogenesis, inflammation, apoptosis, immune response and protein kinase were determined from 12 hours to 3 weeks post-irradiation. Then, 43 temporal clusters corresponding to profiles of similar expressions, including 49 genes out of 301 initially measured, were generated according to the proposed method. Forty-seven transcription factors (TFs) responsible for the expression of clusters of genes were predicted from sequence regulatory elements using the MotifMap system. Their temporal profiles of occurrences were established and clustered. Dynamic network interactions and molecular pathways of TFs and differential genes were finally explored, revealing key node genes and putative important cellular processes involved in tissue infiltration by immune cells following exposure to a radiotherapy dose fraction.


Antibacterial and Antioxidant Properties of the Leaves and Stem Essential Oils of Jatropha gossypifolia L.
Antibacterial and antioxidant properties of the leaves and stem essential oils (EOs) of Jatropha gossypifolia and their efficacies against infectious and oxidative stress diseases were studied in vitro. The EOs obtained using Clevenger modified apparatus were characterized by high resolution GC-MS, while their antioxidant and antibacterial properties were examined by spectrophotometric and agar diffusion techniques, respectively. The EOs exhibited strong antibacterial activity against Escherichia coli, Enterococcus faecium, and Staphylococcus aureus. The stem essential oil (SEO) was more active than the leaf essential oil (LEO) against test bacteria with minimum inhibition concentration (MIC) ranging from 0.025 to 0.05 mg/mL and the LEO from 0.05 to 0.10 mg/mL. The SEO was bactericidal at 0.025 and 0.05 mg/mL against S. aureus and E. faecium, respectively, and the LEO was bacteriostatic against the three bacteria at 0.05 and 0.10 mg/mL. The SEO IC50 (0.07 mg/mL) showed that the antiradical strength was superior to LEO (0.32 mg/mL) and β-carotene (1.62 mg/mL) in scavenging 2, 2-diphenyl-1-picrylhydrazyl radicals (DPPH•). The oils effectively reduced three other oxidants to neutral molecules in concentration dependent manner. Findings from this study suggest that, apart from the traditional uses of the plant extracts, the EOs have strong bioactive compounds with noteworthy antibacterial and antiradical properties and may be good candidates in the search for lead compounds for the synthesis of novel potent antibiotics.


A hyphenated echelle LIBS-Raman system for multi-purpose applications.
We have developed and standardized a novel hybrid laser-induced breakdown spectroscopy (LIBS)-Raman system using a single pulsed laser and a high-resolution intensified charge coupled device coupled echelle spectrograph. LIBS and Raman spectroscopy are highly complementary techniques which yield elemental and molecular information. Both techniques share an apparently similar instrumental configuration but need entirely different requirements like spectral range covered, resolution, and light-gathering efficiencies. There are thus many challenges to be faced in developing a combined system. In the present work, we show that an echelle spectrograph combined with a compact Q-switched Nd:YAG laser operating at 532 nm as an excitation source in a portable configuration can be efficiently used for such multi-purpose spectroscopy. Atomic and molecular emissions from the sample surface have been recorded in a gated mode using this setup. Compared to conventional spectrographs, echelle provides simultaneous broad bandpass (250-900 nm) and better spectral resolution at an extremely small fixed slit width of 10 × 50 μm without moving the dispersive elements. The echelle-based hyphenated system provides fast and reliable analysis of materials with combined atomic and molecular spectra of the same spot with better reliability. In this paper, we discuss the optimization of various instrumental parameters and optical components of this hyphenated system using a medium Raman cross section sample, CaCO3. The feasibility of single shot LIBS-Raman measurement capabilities of echelle has also been demonstrated using the developed system.


Processing of DNA double strand breaks by alternative non-homologous end-joining in hyperacetylated chromatin.
Mammalian cells employ at least two subpathways of non-homologous end-joining for the repair of ionizing radiation induced DNA double strand breaks: The canonical DNA-PK-dependent form of non-homologous end-joining (D-NHEJ) and an alternative, slowly operating, error-prone backup pathway (B-NHEJ). In contrast to D-NHEJ, which operates with similar efficiency throughout the cell cycle, B-NHEJ operates more efficiently in G2-phase. Notably, B-NHEJ also shows strong and as of yet unexplained dependency on growth activity and is markedly compromised in serum-deprived cells, or in cells that enter the plateau-phase of growth. The molecular mechanisms underpinning this response remain unknown. Since chromatin structure or changes in chromatin structure are prime candidate-B-NHEJ-modulators, we study here the role of chromatin hyperacetylation, either by HDAC2 knockdown or treatment with the HDAC inhibitor TSA, on the repair by B-NHEJ of IR-induced DSBs. siRNA-mediated knockdown of HDAC2 fails to provoke histone hyperacetylation in Lig4-/- MEFs and has no detectable effect on B-NHEJ function. Treatment with TSA that inhibits multiple HDACs causes efficient, reversible chromatin hyperacetylation in Lig4-/- MEFs, as well as in human HCT116 Lig4-/- cells and the human glioma cell line M059K. The IR yield of DSBs in TSA-treated cells remains similar to that of untreated cells despite the expected chromatin relaxation. In addition, chromatin hyperacetylation leaves unchanged repair of DSBs by B-NHEJ in irradiated exponentially growing, or plateau-phase cells. Notably, under the experimental conditions employed here, chromatin hyperacetylation fails to detectably modulate B-NHEJ in M059K cells as well. In summary, the results show that chromatin acetylation or deacetylation does not affect the kinetics of alternative NHEJ in all types of cells examined both in exponentially growing and serum deprived cultures. We conclude that parameters beyond chromatin acetylation determine B-NHEJ efficiency in the plateau-phase of growth.


IsdC from Staphylococcus lugdunensis induces biofilm formation under low-iron growth conditions.
Staphylococcus lugdunensis is a coagulase-negative staphylococcus that is a commensal of humans and an opportunistic pathogen. It can cause a spectrum of infections, including those that are associated with the ability to form biofilm, such as occurs with endocarditis or indwelling medical devices. The genome sequences of two strains revealed the presence of orthologues of the ica genes that are responsible for synthesis of poly-N-acetylglucosamine (PNAG) that is commonly associated with biofilm in other staphylococci. However, we discovered that biofilm formed by a panel of S. lugdunensis isolates growing in iron-restricted medium was susceptible to degradation by proteases and not by metaperiodate, suggesting that the biofilm matrix comprised proteins and not PNAG. When the iron concentration was raised to 1 mM biofilm formation by all strains tested was greatly reduced. A mutant of strain N920143 lacking the entire locus that encodes iron-regulated surface determinant (Isd) proteins was defective in biofilm formation under iron-limited conditions. An IsdC-null mutant was defective, whereas IsdK, IsdJ, and IsdB mutants formed biofilm to the same level as the parental strain. Expression of IsdC was required both for the primary attachment to unconditioned polystyrene and for the accumulation phase of biofilm involving cell-cell interactions. Purified recombinant IsdC protein formed dimers in solution and Lactococcus lactis cells expressing only IsdC adhered to immobilized recombinant IsdC but not to IsdJ, IsdK, or IsdB. This is consistent with a specific homophilic interaction between IsdC molecules on neighboring cells contributing to accumulation of S. lugdunensis biofilm in vivo.


Consortium for oral health-related informatics: improving dental research, education, and treatment.
Advances in informatics, particularly the implementation of electronic health records (EHR), in dentistry have facilitated the exchange of information. The majority of dental schools in North America use the same EHR system, providing an unprecedented opportunity to integrate these data into a repository that can be used for oral health education and research. In 2007, fourteen dental schools formed the Consortium for Oral Health-Related Informatics (COHRI). Since its inception, COHRI has established structural and operational processes, governance and bylaws, and a number of work groups organized in two divisions: one focused on research (data standardization, integration, and analysis), and one focused on education (performance evaluations, virtual standardized patients, and objective structured clinical examinations). To date, COHRI (which now includes twenty dental schools) has been successful in developing a data repository, pilot-testing data integration, and sharing EHR enhancements among the group. This consortium has collaborated on standardizing medical and dental histories, developing diagnostic terminology, and promoting the utilization of informatics in dental education. The consortium is in the process of assembling the largest oral health database ever created. This will be an invaluable resource for research and provide a foundation for evidence-based dentistry for years to come.


Enhancing the delivery of erythropoietin and its variants into the ischemic brain.
The hematopoietic growth factor erythropoietin (EPO) and its neuroprotective, but not hematopoietic, variants asialoEPO, carbamylated EPO (CEPO), and low sialic acid EPO (Neuro-EPO) are attractive candidates for stroke treatment. Due to their large molecular weight, these proteins enter the brain only to a minor extent when intravenously administered, which has raised the question for alternative delivery strategies, among which intranasal delivery may certainly be an attractive choice, as the review by Garcia Rodriguez and Sosa Teste in this journal points out. Before this strategy may be considered clinically applicable, however, more and, in particular, quantitative information is needed about (a) the temporospatial accumulation of EPO and its variants in the brain tissue both in animals and nonhuman primates, and (b) the accumulation of EPO and its variants in the human cerebrospinal fluid.


Workplace-Based Assessment in Cross-Border Health Professional Education.
Construct: The globalization of healthcare has been accentuated by the export of health professional curricula overseas. Yet intact translation of pedagogies and practices devised in one cultural setting may not be possible or necessarily appropriate for alternate environments. Purposeful examination of workplace learning is necessary to understand how the source or "home" program may need adapting in the distributed or "host" setting. Background: Strategies to optimize cross-border medical education partnerships have been largely focused on elements of campus-based learning. Determining how host clinical supervisors approach assessment in experiential settings within a different culture and uphold the standards of home programs is relevant given the influence of context on trainees' demonstrated competencies. In this mixed-methods study, we sought to explore assessor judgments of student workplace-based performance made by preceptors sharing a pharmacy curriculum in Canada and Qatar. Approach: Using modified Delphi consensus technique, we asked clinical supervisors in Canada (n = 18) and in Qatar (n = 14) to categorize trainee performance as described in 16 student vignettes. The proportion of ratings for three levels of expectation (exceeds, meets, or below) was calculated and within-country group consensus achieved if the level of agreement reached 80%. Between-country group comparisons were measured using a chi-square statistic. We then conducted follow-up semi-structured interviews to gain further perspectives and clarify assessor rationale. Transcripts were analyzed using thematic content analysis. Results: The threshold for between-country group differences in assessor impressions was met for only two of the 16 student vignettes. Compared to Canadian clinical supervisors, relatively more preceptors in Qatar judged one described student as meets rather than exceeds expectations and one as meets rather than falls below expectations. Analysis of follow-up interviews exploring how culture may inform variations in assessor judgments identified themes associated with the profession, organization, learner, and supervisor performance theories but not their particular geographic context. Clinical supervisors in both countries were largely aligned in expectations of student knowledge, skills, and behaviors demonstrated in patient care and multidisciplinary team interactions. Conclusions: Our study demonstrated that variation in student assessment was more frequent among clinical supervisors within the same national context than any differences identified between the two countries. In these program settings, national sociocultural norms did not predict global assessor impressions or competency-specific judgments; instead, professional and organizational cultures were more likely to inform student characterizations of performance in workplace-based settings. Further study situated within the specific experiential learning contexts of cross-border health professional curricula is assuredly warranted.