Usefulness of a simulated experience method for transfer assistance for hemiplegia or limited range of motion in multiple joints.
[Purpose] To further the understanding of dysfunctions to which a simulated experience method could be applied, we clarified whether a simulated experience method can promote caregivers to utilize the abilities of care recipients with pseudo-hemiplegia or pseudo-limited range of motion (ROM) in multiple joints. [Participants and Methods] We studied transfer assistance in 60 nursing assistants from nursing home settings: 30 were assigned to the pseudo-hemiplegia (26 females, 4 males) and limited ROM in multiple joints (27 females, 3 males) groups. One healthy person was fitted with orthotic braces to mimic hemiplegia or limited ROM in multiple joints, each making it difficult to stand-up. Participants were randomized to either a simulated experience group (involving interventional help from a physical therapist between the first and second measurements) or a control group. The load difference on the lower limbs of the care recipient between two rounds of transfer assistance was examined. [Results] The difference between the second and first measurements was -5.9 ± 74.5 N for the control group and 107.9 ± 123.6 N for the simulated experience method in the pseudo-hemiplegia study, and -14.7 ± 64.7 N and 149.1 ± 132.4 N, respectively, for the pseudo-limited ROM-in-multiple-joints study. [Conclusion] The simulated experience method promoted transfer assistance of a care recipient with pseudo-hemiplegia or pseudo-limited ROM in multiple joints. These results suggest that hemiplegia and limited ROM in multiple joints are added as dysfunctions that can be applied to a simulated experience method in transfer assistance.


Development and implementation of a screen-and-refer approach to addressing maternal depression, substance use, and intimate partner violence in home visiting clients.
Perinatal maternal depression (MD), substance use (SU), and intimate partner violence (IPV) are critical public health concerns with significant negative impacts on child development. Bolstering the capacity of home visiting (HV) programs to address these significant risk factors has potential to improve child and family outcomes. This study presents a description and mixed-methods feasibility evaluation of the "Home Visitation Enhancing Linkages Project (HELP)," a screen-and-refer approach to addressing MD, SU, and IPV within HV aimed at improving risk identification and linkage to treatment among HV clients. HELP was a three-phase intervention that included three evidence-based interventions: screening, motivational interviewing (MI), and case management (CM). This study presents quantitative fidelity data from 21 home visitors reporting on 116 clients in 4 HV programs, as well as qualitative data from structured interviews with 14 home visitors. Nearly all clients were screened and 22% screened positive on at least one risk domain. Rates of MI and CM implementation were lower than expected, however home visitors implemented general supportive interventions at high rates. Home visitor interviews revealed the following factors that may have impacted HELP implementation: client disclosure of risk, barriers to treatment access, systems integration, home visitor role perception, and integration of HELP into the broader HV curriculum. Implications of study findings for the design of future attempts to address maternal risk within HV are discussed.


Thirty-day readmission rate: A predictor of initial surgical severity or quality of surgical care? A regional hospital analysis.
Unplanned readmission within 30 days is currently being used in high-income countries (HICs) for measuring the quality of surgical care. Surgical site infection (SSI), abdominal complaints and pain are the most common causes for such readmission. The correlation between readmission rates and mortality, increased patient volumes and complexity of surgery remains controversial. To explore the risk factors for unplanned readmission in the surgical population in a low- and middle-income country setting. This is a retrospective review of prospectively collected data of unplanned 30-day readmissions from January 2014 to December 2017 in the Department of General Surgery, Worcester Hospital, South Africa (SA). Basic patient demographics, reasons for readmission, operative urgency and severity, wound class, length of stay and final outcomes were used to compare the inpatient cohort and identify predictors for unplanned readmission. A total of 9 649 patients were admitted to the general surgery department at Worcester Hospital - 2.87% (n=270) were unplanned readmissions within 30 days. The mean age of this cohort was 42 (standard deviation (SD) 22) years, with 61% male patients. SSI (60.37%; n=163), gastrointestinal complications (24.44%; n=66) and blood transfusion (7.03%; n=19) were the most common causes for readmission. Median initial length of stay (LOS) was 4 days; after readmission it was 5 days. Readmissions were responsible for 1 914 additional patient days. Operative Portsmouth-POSSUM (P-POSSUM) (Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity) score (p<0.001), increase in operative wound classification (p=0.001) and emergency surgical procedures (p=0.001) were significant risk factors for readmission within 30 days. The Department of General Surgery, Worcester Hospital, had a readmission rate comparable with that in HICs. Readmission rate is an indicator of advanced surgical pathology requiring an operative intervention of greater magnitude, often presenting as an emergency. Our results can be used to improve postoperative surveillance and ultimately improve outcomes in high-risk surgical populations. This study provides a benchmark for other regional hospitals in SA and has implications for quality-improvement programmes.


[Comparison of ethanol and acetaldehyde interactions with chromatin components of cell nuclei from the rat liver].
Interaction of ethanol and acetaldehyde with structures of rat liver cells nuclei was studied in vivo and in vitro using labelled compounds. After injection of 14C-ethanol and 14C-acetaldehyde into control and alcohol-consuming animals the radioactivity was found in carioplasm, non-histone proteins and residual DNA-protein complex. Relative value of specific radioactivity of all the chromatin components studied was practically similar after administration of either ethanol or acetaldehyde. The effect of such aldehyde dehydrogenase inhibitor as teturam led to a significant increase of the label incorporation into chromatin proteins. Acetaldehyde was shown to bind intensively with the isolated chromatin matrix. At the same time, radioactivity was not found in chromatin after incubation with labelled ethanol. The data obtained suggest that alterations in structure and functions of genetic cell apparatus under conditions of ethanol intoxication occurred mainly due to the action of acetaldehyde as a result of ethanol reduction.


CD4+ T cell help is dispensable for protective CD8+ T cell memory against mousepox virus following vaccinia virus immunization.
It has been shown in various infection models that CD4(+) T cell help (TH) is necessary for the conditioning, maintenance, and/or recall responses of memory CD8(+) T cells (CD8M). Yet, in the case of vaccinia virus (VACV), which constitutes the vaccine used to eradicate smallpox and is a candidate vector for other infectious diseases, the issue is controversial because different groups have shown either T(H) dependence or independence of CD8M conditioning, maintenance, and/or recall response. In agreement with some of these groups, we show that T(H) plays a role in, but is not essential for, the maintenance, proliferation, and effector differentiation of polyclonal memory CD8(+) T cells after infection with wild-type VACV strain Western Reserve. More important, we show that unhelped and helped anti-VACV memory CD8(+) T cells are similarly efficient at protecting susceptible mice from lethal mousepox, the mouse equivalent of human smallpox. Thus, T(H) is not essential for the conditioning and maintenance of memory CD8(+) T cells capable of mounting a recall response strong enough to protect from a lethal natural pathogen. Our results may partly explain why the VACV vaccine is so effective. We used vaccinia virus (VACV)--a gold standard vaccine--as the immunogen and ectromelia virus (ECTV) as the pathogen to demonstrate that the conditioning and maintenance of anti-VACV memory CD8(+) T cells and their ability to protect against an orthopoxvirus (OPV) infection in its natural host can develop in the absence of CD4(+) T cell help. Our results provide important insight to our basic knowledge of the immune system. Further, because VACV is used as a vaccine in humans, our results may help us understand how this vaccine induces protective immunity in this species. In addition, this work may partly explain why VACV is so effective as a vaccine.


TP53INP1, a tumor suppressor, interacts with LC3 and ATG8-family proteins through the LC3-interacting region (LIR) and promotes autophagy-dependent cell death.
TP53INP1 (tumor protein 53-induced nuclear protein 1) is a tumor suppressor, whose expression is downregulated in cancers from different organs. It was described as a p53 target gene involved in cell death, cell-cycle arrest and cellular migration. In this work, we show that TP53INP1 is also able to interact with ATG8-family proteins and to induce autophagy-dependent cell death. In agreement with this finding, we observe that TP53INP1, which is mainly nuclear, relocalizes in autophagosomes during autophagy where it is eventually degraded. TP53INP1-LC3 interaction occurs via a functional LC3-interacting region (LIR). Inactivating mutations of this sequence abolish TP53INP1-LC3 interaction, relocalize TP53INP1 in autophagosomes and decrease TP53INP1 ability to trigger cell death. Interestingly, TP53INP1 binds to ATG8-family proteins with higher affinity than p62, suggesting that it could partially displace p62 from autophagosomes, modifying thereby their composition. Moreover, silencing the expression of autophagy related genes (ATG5 or Beclin-1) or inhibiting caspase activity significantly decreases cell death induced by TP53INP1. These data indicate that cell death observed after TP53INP1-LC3 interaction depends on both autophagy and caspase activity. We conclude that TP53INP1 could act as a tumor suppressor by inducing cell death by caspase-dependent autophagy.


A sequence-based 74plex microhaplotype assay for analysis of forensic DNA mixtures.
Microhaplotypes are emerging biomarkers for forensic applications. In this study, a sequence-based multiplex assay of 74 microhaplotypes (230 SNPs) was developed on the Ion Torrent S5™ (Thermo Fisher Scientific) system and the potential for its application to mixture deconvolution was explored. The 74 loci are distributed across the autosomal human genome and have Ae (i.e., effective number of alleles) values ranging from 1.307 to 6.010 (median = 2.706) and In (i.e., informativeness) values ranging from 0.096 to 0.660 (median = 0.251); the amplicon sizes range between 157 and 325 bp. The typing performance of the panel was evaluated on a series of in-silico two to five-person DNA mixtures and results were compared to fragment and sequence-based STRs. The 74plex-locus assay was found sensitive down to 0.05 ng of input DNA and effective for the analysis of mixtures at different contributor ratios and input DNA amounts. As expected, none or very partial minor CE-STR profile(s) were reported for highly imbalanced two-person and high-order DNA mixtures while sequencing of STRs enabled the detection of more individual minor alleles. For microhaplotypes, a full minor profile was detected down to a 20:1 ratio at 10 ng and minimal allele dropout at 1 ng of input DNA. A higher rate of allele dropout from the minor donor(s) was reported at 1 ng than 10 ng for three-person mixtures while for four- and five-person mixtures, the same number of dropouts was observed for almost all minor donors. Overall this microhaplotype panel is a powerful tool that can complement and enhance size- and sequence-based STR analysis of forensic DNA mixtures.


Efficacy of Ceftolozane-Tazobactam in Combination with Colistin against Extensively Drug-Resistant Pseudomonas aeruginosa, Including High-Risk Clones, in an In Vitro Pharmacodynamic Model.
Combination therapy is an attractive therapeutic option for extensively drug-resistant (XDR) Pseudomonas aeruginosa infections. Colistin has been the only treatment available for these infections for many years, but its results are suboptimal. Ceftolozane-tazobactam (C/T) is a newly available therapeutic option that has shown good antipseudomonal activity, even against a number of XDR P. aeruginosa strains. However, data about combinations containing C/T are scarce. The aim of this study was to analyze the activity of C/T and colistin alone and in combination against a collection of XDR P. aeruginosa strains containing 24 representative clinical isolates from a multicentre Spanish study. Twenty-four time-kill experiments performed over 24 h were conducted in duplicate to determine the effects of colistin and C/T alone and combined. An in vitro pharmacodynamic chemostat model then was used to validate this combination against three selected XDR P. aeruginosa ST175 isolates with different susceptibility levels to C/T. Static time-kill assays demonstrated superior synergistic or additive effect for C/T plus colistin against 21 of the 24 isolates studied. In the in vitro dynamic pharmacokinetic/pharmacodynamic (PK/PD) model, the C/T regimen of 2/1 g every 8 h with a steady-state concentration of 2 mg/liter colistin effectively suppressed the bacterial growth at 24 h. Additive or synergistic interactions were observed for C/T plus colistin against XDR P. aeruginosa strains and particularly against C/T-resistant strains. C/T plus colistin may be a useful treatment for XDR P. aeruginosa infections, including those caused by high risk-clones resistant to C/T.


Testing Mean Differences among Groups: Multivariate and Repeated Measures Analysis with Minimal Assumptions.
To date, there is a lack of satisfactory inferential techniques for the analysis of multivariate data in factorial designs, when only minimal assumptions on the data can be made. Presently available methods are limited to very particular study designs or assume either multivariate normality or equal covariance matrices across groups, or they do not allow for an assessment of the interaction effects across within-subjects and between-subjects variables. We propose and methodologically validate a parametric bootstrap approach that does not suffer from any of the above limitations, and thus provides a rather general and comprehensive methodological route to inference for multivariate and repeated measures data. As an example application, we consider data from two different Alzheimer's disease (AD) examination modalities that may be used for precise and early diagnosis, namely, single-photon emission computed tomography (SPECT) and electroencephalogram (EEG). These data violate the assumptions of classical multivariate methods, and indeed classical methods would not have yielded the same conclusions with regards to some of the factors involved.


A serological, parasitological and clinical evaluation of untreated Chagas disease patients and those treated with benznidazole before and thirteen years after intervention.
The etiological treatment of Chagas disease is recommended for all patients with acute or recent chronic infection, but controversies remain regarding the benefit of chemotherapy and interpretations of the parasitological cure after etiological treatment. This study compares the laboratory and clinical evaluations of Chagas disease patients who were diagnosed 13 years earlier. Fifty-eight Chagas disease patients (29 treated with benznidazole and 29 untreated) were matched at the time of treatment based on several variables. Conventional serology revealed the absence of seroconversion in all patients. However, lower serological titres were verified in the treated group, primarily among patients who had the indeterminate form of the disease. Haemoculture performed 13 years after the intervention was positive for 6.9% and 27.6% of the treated and untreated patients, respectively. Polymerase chain reaction tests were positive for 44.8% and 13.8% of the treated and untreated patients, respectively. Patients who presented with the indeterminate form of the disease at the beginning of the study exhibited less clinical progression (17.4%) compared with the untreated group (56.5%). Therefore, this global analysis revealed that etiological treatment with benznidazole may benefit patients with respect to the clinical progression of Chagas disease and the prognosis, particularly when administered to patients with the indeterminate form of the disease.


Antiinflammatory Effects of Budesonide in Human Fetal Lung.
Lung inflammation in premature infants contributes to the development of bronchopulmonary dysplasia (BPD), a chronic lung disease with long-term sequelae. Pilot studies administering budesonide suspended in surfactant have found reduced BPD without the apparent adverse effects that occur with systemic dexamethasone therapy. Our objective was to determine budesonide potency, stability, and antiinflammatory effects in human fetal lung. We cultured explants of second-trimester fetal lung with budesonide or dexamethasone and used microscopy, immunoassays, RNA sequencing, liquid chromatography/tandem mass spectrometry, and pulsating bubble surfactometry. Budesonide suppressed secreted chemokines IL-8 and CCL2 (MCP-1) within 4 hours, reaching a 90% decrease at 12 hours, which was fully reversed 72 hours after removal of the steroid. Half-maximal effects occurred at 0.04-0.05 nM, representing a fivefold greater potency than for dexamethasone. Budesonide significantly induced 3.6% and repressed 2.8% of 14,500 sequenced mRNAs by 1.6- to 95-fold, including 119 genes that contribute to the glucocorticoid inflammatory transcriptome; some are known targets of nuclear factor-κB. By global proteomics, 22 secreted inflammatory proteins were hormonally regulated. Two glucocorticoid-regulated genes of interest because of their association with lung disease are CHI3L1 and IL1RL1. Budesonide retained activity in the presence of surfactant and did not alter its surface properties. There was some formation of palmitate-budesonide in lung tissue but no detectable metabolism to inactive 16α-hydroxy prednisolone. We concluded that budesonide is a potent and stable antiinflammatory glucocorticoid in human fetal lung in vitro, supporting a beneficial antiinflammatory response to lung-targeted budesonide:surfactant treatment of infants for the prevention of BPD.


Pathways to Care for Patients with Bipolar-I Disorder: An Exploratory Study from a Tertiary Care Centre of North India.
Understanding the pathways to psychiatric care is important from a public health perspective. Only a few Indian studies have focused on this, particularly for severe mental disorders. The present study was planned to assess it in patients with Bipolar-I disorder (BD-I). Sixty-four patients with DSM 5 diagnosis of BD-I and their caregivers were included. A semi-structured interview proforma was used to gather information. Psychiatrists were the first care provider in 43.8% of the cases, followed by traditional faith healers (32.8%) and general physician/neurologists (17.2%). The median duration of untreated bipolar disorder (DUB) was 21 days (1 day to 152 months). Relatively long DUB (3.5 ± 3.5 years) was found for 17.2% of the sample. The median duration of the first contact with a psychiatrist was 45 days and the interval between the contact with the first care provider and a psychiatrist was 90 days (1 day to 151 months). At the time of first treatment seeking, 64% of patients and caregivers had poor awareness regarding psychiatric treatment. Patients with BD-I seek help from psychiatrists, faith healers or other medical practitioners for multiple reasons. There is a need to sensitise the community and various service providers about early identification and optimum management of BD-I.


13-year follow-up of the German angioplasty bypass surgery investigation.
The German Angioplasty Bypass Surgery Investigation was designed to compare symptomatic efficacy and safety of percutaneous coronary balloon angioplasty (PTCA) with coronary artery bypass surgery (CABG) in patients with symptomatic multi-vessel disease. This follow-up study was performed to determine the long-term outcome of patients following these interventions. From 1986 to 1991, 359 patients with angina CCS class II-IV, age below 75 years, and coronary multi-vessel disease requiring revascularization of at least two major coronary vessels were recruited at eight German centres and randomized to PTCA or CABG. From 337 patients undergoing the planned procedure, 324 patients could be followed-up (96%). Baseline parameters were identical in both groups, 2.2+/-0.6 vessels were treated in CABG patients, whereas 1.9+/-0.5 vessels were treated in PTCA patients. Thirty-seven per cent of surgical patients received internal mammary artery grafts, while no stents were used in patients undergoing PTCA. At the end of the 13-year follow-up period, the degree of angina, the degree of dyspnea, and the utilization of nitrates were comparable in both groups. With a total number of 76 deaths, Kaplan-Meier analysis revealed a comparable distribution in both groups. Although time to first re-intervention was significantly shorter in the PTCA group, P<0.001, frequencies of re-intervention (CABG, n=94; PTCA, n=136) and crossover rates (CABG to PTCA, n=49; PTCA to CABG, n=51) were comparable in both groups. The results of our 13-year follow-up suggest that in patients with symptomatic multi-vessel disease, both PTCA and CABG are associated with a comparable long-term survival and symptomatic efficacy. How far these results may be altered by developments such as drug-eluting stents or off-pump surgery remains to be determined.


A dark strain in the Fusarium solani species complex isolated from primary subcutaneous sporotrichioid lesions associated with traumatic inoculation via a rose bush thorn.
Fusarium species are hyaline hyphomycetes widely distributed in nature and documented agents of both superficial and systemic infections in humans. In this paper, we report a darkly-pigmented and initially non-sporulating isolate in the Fusarium solani species complex (FSSC) causing a post-traumatic sporotrichoid infection in an otherwise healthy, male patient. Sequencing of multiple loci showed that the isolate represented an otherwise unknown lineage, possibly corresponding to a separate species, within the multi-species F. solani complex. In prolonged culture, the non-sporulating isolate produced revertant wild-type subcultures with typical Fusarium conidiation. This suggests that the original dense, dark, non-sporulating isolate was a host-adapted form selected in vivo for characters compatible with human pathogenicity. The production of such forms by Fusarium species is increasingly recognized now that sequencing has allowed the identification of highly atypical isolates. In vitro antifungal susceptibility of the isolate was investigated against seven conventional and two newly approved antifungal agents. The isolate showed in vitro resistance to amphotericin B, but appeared susceptible to itraconazole and terbinafine. A cure was ultimately achieved with combined terbinafine/itraconazole therapy with prolonged itraconazole follow-up therapy.


Altered brain network centrality in Parkinson's disease patients after deep brain stimulation: a functional MRI study using a voxel-wise degree centrality approach.
After deep brain stimulation (DBS), patients with Parkinson's disease (PD) show improved motor symptoms and decreased verbal fluency, an effect that occurs before the initiation of DBS in the subthalamic nucleus. However, the underlying mechanism remains unclear. This study aimed to evaluate the effects of DBS on whole-brain degree centrality (DC) and seed-based functional connectivity (FC) in PD patients. The authors obtained resting-state functional MRI data of 28 PD patients before and after DBS surgery. All patients underwent MRI scans in the off-stimulation state. The DC method was used to evaluate the effects of DBS on whole-brain FC at the voxel level. Seed-based FC analysis was used to examine network function changes after DBS. After DBS surgery, PD patients showed significantly weaker DC values in the left middle temporal gyrus, left supramarginal gyrus, and left middle frontal gyrus, but significantly stronger DC values in the midbrain, left precuneus, and right precentral gyrus. FC analysis revealed decreased FC values within the default mode network (DMN). This study demonstrated that the DC of DMN-related brain regions decreased in PD patients after DBS surgery, whereas the DC of the motor cortex increased. These findings provide new evidence for the neural effects of DBS on voxel-based whole-brain networks in PD patients.


A cohort study of drug users' compliance with zidovudine treatment.
Determine whether patients who have acquired the human immunodeficiency virus through injecting drug use receive less antiviral medication (zidovudine) than comparable patients of other risk groups considering access to, acceptance of, and compliance with treatment. Historical cohort study. Human immunodeficiency virus outpatient clinic. Human immunodeficiency virus-infected subjects eligible for zidovudine treatment between January 1, 1989, and January 1, 1992, comparing injecting drug users (IDUs) with non-IDUs ("others"). Proposal, acceptance, start of, and compliance with zidovudine treatment. One hundred fifty-one IDUs and 162 other human immunodeficiency virus-positive subjects became eligible for zidovudine treatment between January 1, 1989, and January 1, 1992. Both groups were proposed zidovudine as often, but zidovudine treatment was refused by 14.9% of IDUs compared with 7.1% of others (P = .029). The IDUs needed considerably more time than other subjects to accept zidovudine therapy (median delay between indication and start of zidovudine treatment, 61 days vs 30 days, P = .0001). After accepting, IDUs were as compliant with treatment as others: 81.3% vs 83.2% were good compliers, and rises of mean corpuscular volume of erythrocytes after 3 and 6 months of treatment were similar in both groups. Former drug users and IDUs receiving methadone were started on zidovudine treatment more often and complied better with treatment than active drug users. Absence of housing and presence of psychiatric diagnosis (both more prevalent in IDUs) were associated with less zidovudine treatment and worse compliance. Injecting drug users tend to delay the start of zidovudine treatment. However, once they have started, their compliance is no worse than the compliance of patients from other risk groups. These results have important implications for clinical trials, medical care, and public health.


The roles of contextual and individual social determinants of oral health-related quality of life in Brazilian adults.
This study investigated the relationship between contextual and individual social determinants of oral health-related quality of life (OHRQoL) in Brazilian adults. Data of 4594 Brazilian adults aged 35-44 years from the 2010 Brazilian Oral Health Survey were analysed. OHRQoL was measured using the Oral Impacts on Daily Performance (OIDP). Contextual socio-economic characteristics at city level assessed in 1991 and 2000 included social deprivation measured by Human Development Index (HDI) and income inequality (Gini index). Individual socio-economic characteristics included family income, schooling and number of goods. Covariates were age, sex, ethnicity and clinical oral measures. Multilevel multivariable Poisson regression analysis was carried to investigate the association of contextual and individual social characteristics with OIDP extent and with each OIDP item. Eating and cleaning teeth were the most reported daily activities influenced by oral conditions. In the adjusted analysis, low HDI 1991 and individual social characteristics (lower family income and lower schooling) were associated with OIDP extent. Sex (females) and all clinical oral measures were also associated with OIDP. Adults living in the cities with low HDI 1991 were more likely to have impact on eating and sleeping, whereas living in cities with high Gini index 1991 increased the prevalence of impact on emotional status, work and social contact. Low income was associated with all OIDP items. Poor contextual social determinants and lower individual socio-economic position are associated with worse OHRQoL among Brazilian adults, even after adjusting for individual socio-demographic and clinical oral health variables.


Lesson of the month 2: Use of thrombolysis for ischaemic stroke in pregnancy - a case report and review of literature.
A nine-week pregnant 33-year-old female presented with sudden-onset right-sided hemiparesis, hemisensory loss, dysarthria and homonymous hemianopia. She was known to have eleven previous miscarriages and used recreational drugs. A CT-head was unremarkable. The patient was subsequently diagnosed with an acute thromboembolic infarct and chose alteplase after counselling. During thrombolysis an ultrasound identified a foetus with no concerning features. A post-thrombolysis CT revealed a left-sided posterior cerebral infarct. CT-venography, carotid Doppler, 72-hour Holter monitor, thrombophilia and an autoimmune screen were all normal. A transthoracic echocardiogram demonstrated a mobile intra-atrial septum with a patent foramen ovale confirmed on bubble echocardiogram. Three days post-thrombolysis the patient requested a termination of pregnancy. A subsequent transvaginal ultrasound demonstrated a missed miscarriage for which the patient underwent evacuation of retained products of conception. Multidisciplinary care ensured that her sensorimotor deficit resolved grossly with only mild dysarthria and right-hand fine motor incoordination.


5% imiquimod cream and reflectance-mode confocal microscopy as adjunct modalities to Mohs micrographic surgery for treatment of basal cell carcinoma.
Imiquimod is an immune response modifier that up-regulates cytokines and has been shown in clinical studies to reduce or clear basal cell carcinoma tumors when applied topically. The objectives were to evaluate the efficacy of 5% imiquimod cream in treating basal cell carcinoma preceding excision by Mohs micrographic surgery and to determine if reflectance-mode confocal microscopy is useful to establish the need for surgical intervention after imiquimod treatment. Subjects applied study cream to one biopsy-confirmed basal cell carcinoma tumor 5 x/week for 2, 4, or 6 weeks in this vehicle-controlled, double-blind study. Confocal microscopy was used for the 6-week treatment group to examine the target tumor area at each interval visit and immediately before Mohs micrographic surgery. After the Mohs micrographic surgery excision, the tissue was evaluated histologically, and the excision area was measured. Confocal microscopy readings were correlated to the histologic diagnosis. Tumors cleared or the target tumor area was reduced in subjects in the 4- and 6-week dosing regimens. Confocal microscopy assessments correlated well with the histologic diagnosis. Imiquimod improved excision results relative to vehicle when used for treating basal cell carcinoma before Mohs micrographic surgery. Confocal microscopy assessments correlated well with tumor response to therapy, suggesting that confocal microscopy may help determine the need for surgery.


The alteration of hypoxia inducible factor-1 (HIF-1) and its target genes in mood disorder patients.
Recent studies suggest that the dysfunction of neural plasticity is associated with mood disorders. Hypoxia-inducible factor-1 (HIF-1), which is a transcriptional activator of vascular endothelial growth factor (VEGF), activates the cellular response to hypoxia. HIF-1 is ubiquitously expressed in all cells, including peripheral leukocytes. However, little is known about the role of HIF-1 in mood disorder. In the present study, we investigated the mRNA expression levels of HIF-1 (α and β) and its target genes (VEGF, GLUT1, PGK1, PFKFB3, and LDHA) in the peripheral white blood cells of patients with major depressive disorder (MDD) and bipolar disorder (BPD). We found increased expression of HIF- 1α and HIF-1β mRNA, as well as the target genes, VEGF, and PFKFB3 in both MDD and BPD patients in a depressive state compared to healthy control subjects. Furthermore, the mRNA expression levels of GLUT1, PGK1, and LDHA were increased in MDD patients in a depressive state compared to healthy control subjects. We also found increased expression of HIF-1α and LDHA mRNA in MDD patients in a remissive state, whereas the mRNA expression levels of other genes in a remissive state were comparable to those in healthy control subjects. There was no significant difference in mRNA expression levels of the genes examined among patients receiving any type of antidepressant or mood stabilizer. Our data suggest that altered expression of HIF-1 and its target genes mRNA in peripheral blood cells are associated-mainly in a state-dependent manner-with mood disorders (especially with MDD). In addition, altered expression of HIF-1 and its target genes may be associated with the pathophysiology of depression.


Hepatitis B virus genotype distribution among chronic hepatitis B virus carriers in Shanghai, China.
Hepatitis B virus (HBV) genotype distribution is still unclear in China, where a high prevalence of HBV infection exists, although it is well known that HBV can be classified into six genotypes based on intergroup divergence. The aim of this study was to investigate the epidemiological distribution of HBV genotypes and to clarify further the genotype-related differences in the pathogenicity of HBV. Seminested PCR and restriction fragment length polymorphism analysis were conducted in 97 asymptomatic HBV carriers (ASC) and 46 chronic hepatitis (CH), 37 liver cirrhosis (LC) and 44 hepatocellular carcinoma (HCC) patients in Shanghai, China. Two hundred and twenty samples (98.2%) were positive for HBV DNA, and of these, 3 (1.4%), 38 (17.2%) and 179 (81.4%) were classified as genotype A, B and C, respectively. There was a statistically significant difference in the distribution of genotypes B and C among various categories of liver diseases (p < 0.01). The distribution of genotype C showed an increasing trend from ASC, CH and LC to the HCC group; in contrast, the distribution of genotype B showed a decreasing trend in the same order. HBeAg positivity was higher in genotype C than in genotype B in all the subjects or in the ASC group alone (p < 0.05, p < 0.01, respectively). More severe liver damage and a higher mean age were observed in genotype C than in genotype B (p < 0.01, p < 0.05, respectively). These results indicate the following: (1) genotypes A, B and C of HBV exist in Shanghai, China; (2) genotype C is the major genotype in this area; (3) genotype C is associated with the development of severe liver diseases, and (4) genotype B has a relatively good prognosis.


Azoreductase activity of dye-decolorizing bacteria isolated from the human gut microbiota.
The gut microbiota enriches the human gene pool and contributes to xenobiotic metabolism. Microbial azoreductases modulate the reduction of azo-bonds, activating produgs and azo polymer-coated dosage forms, or degrading food additives. Here, we aimed to screen the healthy human gut microbiota for food colorant-reducing activity and to characterize factors modulating it. Four representative isolates from screened fecal samples were identified as E. coli (AZO-Ec), E. faecalis (AZO-Ef), E. avium (AZO-Ev) and B. cereus (AZO-Bc). Both AZO-Ef and AZO-Ev decolorized amaranth aerobically and microaerophilically while AZO-Ec and AZO-Bc had higher aerobic reduction rates. The isolates varied in their activities against different dyes, and the azo-reduction activity mostly followed zero-order reaction kinetics, with a few exceptions. Additionally, the isolates had different pH dependence, e.g., AZO-Ec was not affected by pH variation while AZO-Bc exhibited variable degradation kinetics at different pH levels. Cell-free extracts showed NADH-dependent enzymatic activities 14-19 times higher than extracellular fractions. FMN did not affect the reducing activity of AZO-Ef cell-free extract, whereas AZO-Ec, AZO-Ev and AZO-Bc had significantly higher reduction rates in its presence (P values = 0.02, 0.0001 and 0.02, respectively). Using Degenerate primers allowed the amplification of azoreductase genes, whose sequences were 98-99% similar to genes encoding FMN-dependent-NADH azoreductases.


Endocrine and neuroendocrine host-parasite relationships.
The relationships between parasites and hosts are complex, with many of these interactions involving an amazing degree of biochemical coevolution and communication. Hormones, neurohormones, and growth factors figure prominently in these relationships. In vertebrate hosts, many parasites secrete hormones, neuropeptides, or cytokine-like molecules that influence the host's physiological and immunological responses. Alternatively, the parasites secrete factors that alter the host's hormone levels. Simultaneously, molecules emanating from the host strongly influence the parasites' success. In some cases the host's hormones directly influence the parasites; in others, effects are mediated indirectly via the host's immune system. In invertebrates, the presence of parasites likewise has a major influence on the host's endocrine status and the normal suite of processes governed by hormones, including host development, metamorphosis, and reproduction. In insects, interactions involving juvenile hormone and ecdysteroids are especially well-documented, and recent evidence suggests that neuropeptides may also be affected by parasitism. Moreover, recent data suggest that in some species, such as snails, the host's nervous, neuroendocrine, and immune systems are functionally linked, similar to the complex interactions seen in vertebrates.


The Collagen-Based Medical Device MD-Tissue Acts as a Mechanical Scaffold Influencing Morpho-Functional Properties of Cultured Human Tenocytes.
Mechanotransduction is the ability of cells to translate mechanical stimuli into biochemical signals that can ultimately influence gene expression, cell morphology and cell fate. Tenocytes are responsible for tendon mechanical adaptation converting mechanical stimuli imposed during mechanical loading, thus affecting extracellular matrix homeostasis. Since we previously demonstrated that MD-Tissue, an injectable collagen-based medical compound containing swine-derived collagen as the main component, is able to affect tenocyte properties, the aim of this study was to analyze whether the effects triggered by MD-Tissue were based on mechanotransduction-related mechanisms. For this purpose, MD-Tissue was used to coat Petri dishes and cytochalasin B was used to deprive tenocytes of mechanical stimulation mediated by the actin cytoskeleton. Cell morphology, migration, collagen turnover pathways and the expression of key mechanosensors were analyzed by morphological and molecular methods. Our findings confirm that MD-Tissue affects collagen turnover pathways and favors cell migration and show that the MD-Tissue-induced effect represents a mechanical input involving the mechanotransduction machinery. Overall, MD-Tissue, acting as a mechanical scaffold, could represent an effective medical device for a novel therapeutic, regenerative and rehabilitative approach to favor tendon healing in tendinopathies.


Bcl-2 antagonist killer 1 (BAK1) polymorphisms influence the risk of developing autoimmune rheumatic diseases in women.
Bcl-2 antagonist killer 1 (BAK1) is a Bcl-2 family proapoptotic member suggested as a candidate gene for autoimmune diseases. The influence of BAK1 polymorphisms on the risk of developing autoimmune rheumatic diseases (AIRDs) in women was investigated. A total of 719 Colombian women were included in the present study: 209 had systemic lupus erythematosus, 99 primary Sjögren syndrome, 159 rheumatoid arthritis and 252 were healthy matched controls. Tag single nucleotide polymorphisms (SNPs) and potentially functional variants were typed by TaqMan allele discrimination assays. HLA-DRB1 and HLA-DQB1 typing was performed by reverse dot-blot hybridisation and linkage disequilibrium (LD) with BAK1 SNPs was assessed. SNPs rs513349 (odds ratio (OR) 0.57, 95% CI 0.46 to 0.72, p = <0.001) and rs5745582 (OR 1.61, 95% CI 1.26 to 2.04, p = <0.001) were associated with the AIRDs included in this study. There was a significant increase of the rs513349G-rs561276C-rs5745582A (GCA) haplotype in each patient cohort as compared to controls (OR 1.95, 95% CI 1.50 to 2.54, p = <0.001). These SNPs were not in LD with HLA-DRB1 or HLA-DQB1 genes. The results indicate that the BAK1 polymorphisms influence the risk of acquiring AIRDs in the population studied and are consistent with the paradigm that autoimmune diseases are likely to share common susceptibility variants.


[The age dependence of traumatically induced bone remodeling as studied in the bone scintigram].
Two independent groups of patients were studied by bone scintigraphy to demonstrate age-dependent changes in the appearance time of fractures: 83 patients with 123 fractures of the spine showed neither by multiple regression analysis nor by separation into two subgroups (younger and older than 55 years) a clinically significant difference by scintimetric evaluation. A second group of 162patients with multiple trauma demonstrated no age dependence of the rate of additional bone lesions shown by scintigraphy. Instead a clear dependence on the site of the lesion and the time after trauma was found: fractures of the shafts of long bones and fractures of the pelvis and spine showed up significantly later than fractures in the neighbourhood of the distal joints. 12-14 days after a trauma all fractures may be detected with a high accuracy independently from the fracture site (except for fractures of the skull which did not show up at all in most cases). Thus, the differences in the appearing time of fractures are not age- but site-dependent.


Molecular Basis for Environment Sensing by a Nucleoid-Structuring Bacterial Protein Filament.
The histone-like nucleoid structuring (H-NS) protein controls the expression of hundreds of genes in Gram-positive bacteria through its capability to coat and condense DNA. This mechanism requires the formation of superhelical H-NS protein filaments that are sensitive to temperature and salinity, allowing H-NS to act as an environment sensor. We use multiscale modeling and simulations to obtain detailed insights into the mechanism of H-NS filament's sensitivity to environmental changes. Through the simulations of the superhelical H-NS filament, we reveal how different environments induce heterogeneity of H-NS monomers. Further, we observe that transient self-association within the H-NS filament creates temperature-inducible strain and might mildly oppose DNA binding. We also probe different H-NS-DNA complex architectures and show that complexation enhances the stability of both DNA and H-NS superhelices. Overall, our results provide unprecedented molecular insights into the environmental sensing and DNA interactions of a prototypical nucleoid-structuring bacterial protein filament.


The pathophysiological role of PEDF in bone diseases.
First discovered in 1991 as a factor secreted by retinal pigment epithelial cells, the potency of pigment epithelium derived factor (PEDF) as an anti-angiogenic has led to examination of its role in active bone growth, repair and remodelling. In the musculoskeletal system, PEDF expression occurs particularly at sites of active bone formation. Expression has been noted in osteoblasts and to a lesser degree osteoclasts, the major classes of bone cells. In fact, PEDF is capable of inducing differentiation of precursor cells into mature osteoblasts. Expression and localisation are closely linked with that of vascular endothelial growth factor (VEGF). Studies at the epiphyseal plate have revealed that PEDF expression plays a key role in endochondral ossification, and beyond this may account for the epiphyseal plate's innate ability to resist neoplastic cell invasion. Collagen-1, the major protein in bone, is avidly bound by PEDF, implicating an important role played by this protein on PEDF function, possibly through MMP-2 and -9 activity. Surprisingly, the role of PEDF has not been evaluated more widely in bone disorders, so the challenge ahead lies in a more diverse evaluation of PEDF in various osteologic pathologies including osteoarthritis and fracture healing.


Age changes in neuromuscular junctions of masseter muscle.
There is little information regarding the relationship between aging and ensuing morphological changes in the neuromuscular junction (NMJ) of the masticatory muscle. The masseter of C57BL/6J male mice at three different ages (young adult, 6 months; mature adult, 12 months; and old, 30 months) was studied. Morphological measurements were taken from zinc iodide osmium stained NMJ. Camera lucida drawings were superimposed on a computer monitor via a video camera and traced using a digitizer. The data were treated statistically using multivariate analysis of variance (MANOVA). No difference was found between 6 and 12 months of age, but a significant decrease in morphological parameters in NMJ from 30-month-old animals was found when compared with mature adult animals. Nerve terminal areas, perimeters, longitudinal extent lengths, and fiber diameters were reduced by 24%, 21%, 15.5% and 23%, respectively. Nerve terminal branches and incidence of sprouts were significantly increased in older animals. Age changes in the NMJ morphology are probably associated with altered balance between degeneration and regeneration of nerve terminals. This view is supported by increased terminal sprouting in old mice which is indicative of the plasticity or remodelling of NMJ during aging.


Affinity isolation and characterization of cytochrome P450 102 (BM-3) from barbiturate-induced Bacillus megaterium.
Cytochrome P450 102 (BM-3) is a catalytically self-sufficient enzyme from Bacillus megaterium that is presently accepted as an important model of the mammalian microsomal P450 monooxygenase system. We have developed a novel affinity approach to purify P450 102 in a single chromatographic step and have studied the spectroscopic, catalytic, nucleotide binding, and crystallization properties of the highly purified enzyme. B. megaterium ATCC 14581 was grown to high cell density, and P450 102 was purified rapidly and in high yield by chromatography on adenosine-2',5'-diphosphate agarose from crude cell-free extract. The cytochrome bound to the column with remarkable avidity, in contrast to the significantly weaker binding observed for NADPH-cytochrome P450 reductase. Chromatographic behavior also showed that the cytochrome bound NADP(+)-type nucleotides more tightly than any other cellular polypeptide. The purified protein was electrophoretically homogeneous and had essentially theoretical contents of FAD, FMN, and heme. Optical spectra showed the expected heme and flavin absorption bands, and three previously undescribed charge-transfer-type absorptions were characterized. Molar extinction coefficients in the oxidized, fully reduced, and ferrous carbonyl states have been determined; notable is the large soret extinction in the ferrous carbonyl state (epsilon 449 nm = 143,500 M-1 cm-1). Final preparations were active in the oxidation of a wide variety of substrates. Of the C14 alkyl compounds studied, tetradecyltrimethylammonium bromide showed the highest substrate-dependent oxidation of NADPH, followed by myristate and myristyl alcohol; however, myristate exhibited the lowest Km value. Activities were tightly coupled to NADPH oxidation (> 97%). Phenobarbital, benzphetamine, cocaine, cyclohexane, methanol, ethanol, retinoic acid, benzoate, heptaflourobutyrate, and 7-ethoxycoumarin were not substrates. NADP+ titrations showed, as expected, that the coenzyme was bound very tightly, with an average Kd of 580 nM. Our preparations of P450 102 are of sufficient purity and stability that crystals of the native holoenzyme have been grown.


TERT promoter mutations are a rare event in gastrointestinal stromal tumors.
Recently, the impact of telomere dysregulation on malignant progression has been reported in many cancers. A few studies have examined TERT promoter mutations in gastrointestinal stromal tumors (GISTs). Irregular telomerase activation can be maintained by TERT hot spot alterations and alternative lengthening of telomeres (ALT) characterized by inactivation of either the alpha-thalassemia/mental retardation syndrome X-linked (ATRX) or death domain-associated protein (DAXX). To elucidate the clinicopathological impact of telomere dysregulation in GISTs, we examined 92 cases of GISTs for TERT promoter hot spot mutations along with immunohistochemical analysis of ATRX and DAXX expression, and compared these findings with the clinicopathological features. Univariate clinicopathological analysis revealed that tumor site, smaller tumor size, presence of necrosis, higher mitotic rate (>5/50 high-power fields) and risk classification were prognostic factors for either disease-free survival or overall survival. Two of 92 informative cases (2.2 %) were found to have heterozygous TERT promoter mutations (C228T), and these mutations occurred in a low-risk and a high-risk tumor, respectively. On immunohistochemical analysis for ATRX and DAXX, 16 (17.4 %) and 3 (3.3 %) of 92 cases showed loss of expression of ATRX and DAXX, respectively. Loss of expression of ATRX and DAXX were mutually exclusive except for one case. TERT promoter mutations were also mutually exclusive of the ALT phenotype. Telomere dysregulation was not associated with patient survival; however, telomere dysregulation was frequently observed in tumors of extra-gastric origin, which have an adverse outcome compared to those of gastric origin.


[Opportunities in the pharmaceutical industry and the local development logic based on the Brazilian biomes: the basis for a national policy discussion].
This study discusses new concepts in the technological development of plant-derived medicines and their relevance to public health, highlighting opportunities in the pharmaceutical market both for the production of herbal medicines and the development of new drugs based on plant substrates and molecules. Recent theoretical and scientific premises for the socioeconomic development of the "Knowledge Age" (considering local production arrangements and systems) could guarantee a competitive advantage for the sector, given the exuberant flora and biodiversity in the various Brazilian biomes.


Bowel functions, fecal unconjugated primary and secondary bile acids, and colonic transit in patients with irritable bowel syndrome.
There is an unclear relationship among bowel symptoms, excretion of unconjugated fecal bile acid (UBA), and colonic transit in irritable bowel syndrome (IBS). We measured total and main individual UBA in fecal samples of patients with IBS and assessed relationships among stool frequency or consistency, fecal UBA (total and individual), and colonic transit. In this study 30 healthy volunteers (controls), 31 subjects with IBS with diarrhea (IBS-D), and 30 with IBS with constipation (IBS-C) were placed on 4-day diets containing 100 g fat; we measured stool characteristics, total fecal UBA and fat levels, and overall colonic transit. We assessed univariate associations of total and individual levels of fecal UBA with phenotype (controls, IBS-D, IBS-C) by using the Kruskal-Wallis test; associations between end points were assessed by using Spearman correlations. With response surface regression models, we assessed relationships between stool, colonic transit, and fecal total and secretory UBA. There was a significant association between total fecal UBA and phenotype (P = .029); the association was greater for IBS-D than IBS-C, compared with controls. Fecal levels of primary UBAs (cholic and chenodeoxycholic acids) were higher in subjects with IBS-D, compared with controls (both P < .01). Levels of fecal secretory UBAs (chenodeoxycholic acid, P = .019; deoxycholic acid, P = .025) were lower in subjects with IBS-C compared with controls, whereas levels of the nonsecretory UBA, lithocholic acid, were higher (P = .020). There were significant univariate associations between stool number and form and total fecal UBA (including percentages of lithocholic acid, chenodeoxycholic acid and cholic acid), fecal fat, and colonic transit at 24 and 48 hours after eating. In the regression models, the relative contribution of colonic transit was consistently greater and largely independent of the contribution of bile acids. Measurements of individual UBAs identify changes associated with stool characteristics in patients with IBS; these effects are independent of the effects of colonic transit.


Effectiveness of a family-centered method for the early identification of social-emotional and behavioral problems in children: a quasi experimental study.
Social-emotional and behavioral problems are common in childhood. Early identification of these is important as it can lead to interventions which may improve the child's prognosis. In Dutch Preventive Child Healthcare (PCH), a new family-centered method has been implemented to identify these problems in early childhood. Its main features are consideration of the child's developmental context and empowerment of parents to enhance the developmental context. In a quasi-experimental study, embedded in routine PCH in the Netherlands, regions in which the family-centered method has been implemented (intervention condition) will be compared to "care as usual" regions (control condition). These regions are comparable in regard to socio-demographic characteristics. From more than 3,500 newborn babies, 18-month follow-up data on social-emotional and behavioral development will be obtained. PCH professionals will assess development during each routine well-child visit; participating parents will fill in standardized questionnaires.Primary outcomes in the study are the proportion of social-emotional and behavioral problems identified by PCH professionals in children aged 2-14 and 18 months in both conditions, and the proportion of agreement between the assessment of PCH professionals and parents. In addition, the added value of the family-centered approach will be assessed by comparing PCH findings with standardized questionnaires. The secondary outcomes are the degree to which the needs of parents are met and the degree to which they are willing to disclose concerns. The family-centered method seems promising for early identification of social-emotional and behavioral problems. The results of this study will contribute to evidence-based public health. NTR2681.


Shaping innovations in long-term care for stroke survivors with multimorbidity through stakeholder engagement.
Stroke, like many long-term conditions, tends to be managed in isolation of its associated risk factors and multimorbidity. With increasing access to clinical and research data there is the potential to combine data from a variety of sources to inform interventions to improve healthcare. A 'Learning Health System' (LHS) is an innovative model of care which transforms integrated data into knowledge to improve healthcare. The objective of this study is to develop a process of engaging stakeholders in the use of clinical and research data to co-produce potential solutions, informed by a LHS, to improve long-term care for stroke survivors with multimorbidity. We used a stakeholder engagement study design informed by co-production principles to engage stakeholders, including service users, carers, general practitioners and other health and social care professionals, service managers, commissioners of services, policy makers, third sector representatives and researchers. Over a 10 month period we used a range of methods including stakeholder group meetings, focus groups, nominal group techniques (priority setting and consensus building) and interviews. Qualitative data were recorded, transcribed and analysed thematically. 37 participants took part in the study. The concept of how data might drive intervention development was difficult to convey and understand. The engagement process led to four priority areas for needs for data and information being identified by stakeholders: 1) improving continuity of care; 2) improving management of mental health consequences; 3) better access to health and social care; and 4) targeting multiple risk factors. These priorities informed preliminary design interventions. The final choice of intervention was agreed by consensus, informed by consideration of the gap in evidence and local service provision, and availability of robust data. This shaped a co-produced decision support tool to improve secondary prevention after stroke for further development. Stakeholder engagement to identify data-driven solutions is feasible but requires resources. While a number of potential interventions were identified, the final choice rested not just on stakeholder priorities but also on data availability. Further work is required to evaluate the impact and implementation of data-driven interventions for long-term stroke survivors.


The Severe Respiratory Insufficiency Application (SRI App): a pilot randomised controlled trial.
An application (app) for the Severe Respiratory Insufficiency Questionnaire (SRI) has been designed and developed for mobile devices. In a randomised controlled trial comprising 60 patients with chronic respiratory failure, the app was compared with the classic paper SRI. Thereby, it was shown that the SRI app is a practical tool that is well accepted. Missing values can be completely avoided by using the SRI app. Finally, reliability, convergent and discriminant validities were established. Accordingly, for the individual SRI subscales, Cronbach's alpha ranged between 0.56 and 0.81 (app) and between 0.54 and 0.83 (paper), respectively. The multilingual SRI app is accessible free of charge for non-profit research purposes.


Inheritance and physiological basis for 2,4-D resistance in prickly lettuce (Lactuca serriola L.).
Experiments were conducted to determine the inheritance and physiological basis for resistance to the synthetic auxinic herbicide (2,4-dichlorophenoxy)acetic acid (2,4-D) in a prickly lettuce biotype. Inheritance of 2,4-D resistance in prickly lettuce is governed by a single codominant gene. Absorption and translocation were conducted using (14)C-2,4-D applied to 2,4-D-resistant and -susceptible biotypes. At 96 h after treatment (HAT), the resistant biotype absorbed less applied 2,4-D and retained more 2,4-D in the treated portion of the leaf compared to the susceptible biotype. The resistant biotype translocated less applied 2,4-D to leaves above the treated leaf and crown at 96 HAT compared to the susceptible biotype. No difference in the rate of metabolism of 2,4-D was observed between the two biotypes. Resistance to 2,4-D appears to originate from a reduced growth deregulatory and overstimulation response compared to the susceptible biotype, resulting in lower translocation of 2,4-D in the resistant prickly lettuce biotype.


Detection of Fusion Genes to Determine Minimal Residual Disease in Leukemia Using Next-Generation Sequencing.
Measuring minimal residual disease (MRD), the persistence of leukemic cells after treatment, is important for monitoring leukemia recurrence. The current methods for monitoring MRD are flow cytometry, to assess aberrant immune phenotypes, and digital droplet PCR (ddPCR), to target genetic aberrations such as single-nucleotide variants and gene fusions. We present the performance of an RNA-based next-generation sequencing (NGS) method for MRD gene fusion detection compared with ddPCR. This method may have advantages, including the capacity to analyze different genetic aberrations and patients in 1 experiment. In particular, detection at the RNA level may be highly sensitive if the genetic aberration is highly expressed. We designed a probe-based NGS panel targeting the breakpoints of 11 fusion genes previously identified in clinical patients and 2 fusion genes present in cell lines. Blocking probes were added to prevent nonspecific enrichment. Each patient RNA sample was diluted in background RNA, depleted for rRNA and globin mRNA, converted to cDNA, and prepared for sequencing. Unique sequence reads, identified by unique molecular identifiers, were aligned directly to reference transcripts. The same patient and cell-line samples were also analyzed with ddPCR for direct comparison. Our NGS method reached a maximum sensitivity of 1 aberrant cell in 10 000 cells and was mostly within a factor of 10 compared with ddPCR. Our detection limit was below the threshold of 1:1000 recommended by European Leukemia Net. Further optimizations are easy to implement and are expected to boost the sensitivity of our method to diagnostically obtained ddPCR thresholds.


Meniere's, migraine, and motion sickness.
Elevated Motion Sickness Susceptibility (MSS) in Meniere?s disease (MD) is likely to be a consequence of the onset of MD and not migraine per se. Pathologies of the vestibular system influence MSS. Bilateral vestibular deficits lower MSS, vestibular neuritis or benign paroxysmal positional vertigo have little overall effect, whereas vestibular migraine elevates MSS. However, less is known about MSS in MD, a condition in which many patients experience vestibular loss and migraine symptoms. The authors conducted an online survey that posed diagnostic and disease questions before addressing frequency of headaches, migraines, visual display dizziness (VDD), syncope, social life, and work impact of dizziness (SWID4) and motion sickness susceptibility (MSSQ). The two groups were: diagnosed MD individuals with hearing loss (n = 751) and non-MD individuals in the control group (n = 400). The MD group showed significantly elevated MSS, more headache and migraine, increased VDD, higher SWID4 scores, and increased syncope. MSS was higher in MD than controls only after the development of MD, but not before, nor in childhood. Although elevated in MD compared with controls, MSS was lower than migraine patients from past data. Multivariate analysis revealed VDD, SWID4, and MSS in adulthood as the strongest predictors of MD, but not headache nor migraine.


Electron temperature and density probe for small aeronomy satellites.
A compact and low power consumption instrument for measuring the electron density and temperature in the ionosphere has been developed by modifying the previously developed Electron Temperature Probe (ETP). A circuit block which controls frequency of the sinusoidal signal is added to the ETP so that the instrument can measure both T(e) in low frequency mode and N(e) in high frequency mode from the floating potential shift of the electrode. The floating potential shift shows a minimum at the upper hybrid resonance frequency (f(UHR)). The instrument which is named "TeNeP" can be used for tiny satellites which do not have enough conductive surface area for conventional DC Langmuir probe measurements. The instrument also eliminates the serious problems associated with the contamination of satellite surface as well as the sensor electrode.


A mouse model of proliferative vitreoretinopathy induced by dispase.
A proliferative vitreoretinopathy-like condition induced by intravitreal dispase injection in C57BL/6J mice was studied using ophthalmoscopic and histochemical procedures. The frequency of intravitreal hemorrhage, intravitreal spots, retinal folds and epiretinal membranes was scored by ophthalmoscopic examination at 1, 2, 4, 6 and 8 weeks after the injection. Intravitreal spots corresponded to free cells exhibiting F4/80 immunoreactivity, a macrophage/microglial marker. Retinal folds always appeared before an epiretinal membrane could be observed. Dispase-injected eyes always showed a much higher frequency of folds and membranes than saline-injected eyes. Folds and membranes appeared earlier and were more extensive in the presence of intravitreal hemorrhage than in its absence. Müller retinal cells exhibited significant changes in glial fibrillary acidic protein-immunoreactivity. This was absent in normal Müller cells but, in dispase-injected animals, it was expressed in radial processes at the site of retinal folds, later extending to the whole retina. Both epi- and subretinal membranes contained cells probably derived from Müller cells, since they exhibited co-localization of glial fibrillary acidic protein- and glutamine synthase immunoreactivities. F4/80 was also present in numerous cells within the retina, epi- and subretinal membranes. By contrast, the retinal pigment epithelium cell marker RPE65 was restricted to subretinal membranes. It can be concluded that dispase induced a proliferative vitreoretinopathy-like condition in mice, with a strong contribution of macrophage- and glial-derived cells.


Malaria-related beliefs and behaviour in southern Ghana: implications for treatment, prevention and control.
A research infrastructure was established in two ecological zones in southern Ghana to study the variables of malaria transmission and provide information to support the country's Malaria Action Plan (MAP) launched in 1992. Residents' beliefs and practices about causes, recognition, treatment and prevention of malaria were explored in two ecological zones in southern Ghana using epidemiological and social research methods. In both communities females constituted more than 80% of caretakers of children 1-9 years and the illiteracy rate was high. Fever and malaria, which are locally called Asra or Atridi, were found to represent the same thing and are used interchangeably. Caretakers were well informed about the major symptoms of malaria, which correspond to the current clinical case definition of malaria. Knowledge about malaria transmission is, however, shrouded in many misconceptions. Though the human dwellings in the study communities conferred no real protection against mosquitoes, bednet usage was low while residents combatted the nuisance of mosquitoes with insecticide sprays, burning of coils and herbs, which they largely considered as temporary measures. Home treatment of malaria combining herbs and over-the-counter drugs and inadequate doses of chloroquine was widespread. There is a need for a strong educational component to be incorporated into the MAP to correct misconceptions about malaria transmission, appropriate treatment and protection of households. Malaria control policies should recognize the role of home treatment and drug shops in the management of malaria and incorporate them into existing control strategies.


The Relation Between the Emergence of Fluoroquinolone Resistance and Fluoroquinolone Exposure in New Cases of Active Pulmonary Tuberculosis.
This study aimed to determine the ratio of fluoroquinolone (FQ) exposure before the diagnosis of patients with a new case of active pulmonary tuberculosis (TB) and to investigate the correlation of this treatment with the emergence of FQ-resistant strains. In this retrospective comparative case series study, a total of 132 patients, who had been diagnosed with adult, culture-positive, active pulmonary TB were reviewed. The FQ group had 30 patients who had had ≥1 time and ≥7 days of FQ exposure within 1 year before the diagnoses. The control group included an equal number of patients with TB with similar demographic characteristics (non-FQ group). Ofloxacin (OFX) and moxifloxacin (MFX) resistance were examined at 2 different concentrations (2 and 4 mg/L for OFX; 0.25 and 0.5 mg/L for MFX). Of the 132 patients, 30 (22%) had 7 days or longer of FQ monotherapy within 1 year of initiation of anti-TB treatment. FQ resistance was detected in 2 (3.3%) patients. In the FQ group, MFX resistance at 0.25 mg/L concentration was observed in 1 patient, whereas another patient had OFX and MFX resistance at 4 mg/L and 0.5 mg/L concentrations, respectively. In the non-FQ group, no FQ resistance was detected in any of the patients. No statistically significant difference in terms of development of FQ resistance was found between the ratios of FQ and non-FQ groups (p=0.492). Although there was no statistically significant difference, 2 patients, in whom resistance was detected, had FQ exposure before their diagnosis. The FQ exposure ratio before the diagnosis is high (22%) in this cohort that includes patients with new active pulmonary TB, and the presence of patients with FQ resistance (even if only a few) should be a noteworthy and cautionary result in terms of FQ exposure and resistance development.


The effect of grinding and/or airborne-particle abrasion on the bond strength between zirconia and veneering porcelain: a systematic review.
Objective: The aim of the study was to make an inventory of current literature on the bond strength between zirconia and veneering porcelain after surface treatment of zirconia by grinding with diamond bur and/or with airborne-particle abrasion. Material and methods: The literature search for the present review was made following recommended guidelines using acknowledged methodology on how to do a systematic review. The electronic databases PubMed, Cochrane Library, and Science Direct were used in the present study. Results: Twelve studies were selected. Test methods used in the original studies included shear bond strength (SBS) test, tensile bond strength test, and micro-tensile bond strength test. The majority of studies used SBS. Results showed a large variation within each surface treatment of zirconia, using different grain size, blasting time, and pressure. Conclusions: Airborne-particle abrasion might improve the bond strength and can therefore be considered a feasible surface treatment for zirconia that is to be bonded. Grinding has been recommended as a surface treatment for zirconia to improve the bond strength; however, this recommendation cannot be verified. A standardized test method and surface treatment are required to be able to compare the results from different studies and draw further conclusions.


Endogenous viral gene ev21 is not responsible for the expression of late feathering in chickens.
The late-feathering (LF) gene K on the Z chromosome is an important gene in the chicken industry, which is frequently utilized for the feather sexing, a type of autosexing, of neonatal chicks. The K gene is closely associated with the endogenous ev21 gene from an avian leukosis virus and the incomplete duplication (ID) of prolactin receptor (PRLR) and sperm flagellar protein 2 (SPEF2) genes, and ev21 has been used as a molecular marker to detect LF birds. In the present study, a comprehensive survey for the presence or absence of ev21 and ID across 1,994 birds from 52 chicken breeds, three commercial hybrid groups, and the Red Jungle Fowl revealed that almost all LF breeds have both ev21 and ID. However, only one LF breed (Ingie) has only ID and no ev21. Moreover, this study revealed that almost all early (normal)-feathering (EF) breeds lack both ev21 and ID, but only one breed (White Plymouth Rock) included EF birds with ev21 but no ID. Therefore, regarding LF expression, the results indicated that ID is responsible, but ev21 is not required. Henceforth, ID should be used as a molecular marker to detect LF birds instead of ev21. Because ev21 contains the full genome of an avian leukosis virus, there is a risk of disease development in breeds with this gene. Therefore, the Ingie breed, which has no ev21 at the K locus, represents excellent material for the establishment of new LF stocks.


Patterns of cortical thinning in the language variants of frontotemporal lobar degeneration.
Frontotemporal lobar degeneration (FTLD) is a clinically, genetically, and pathologically heterogeneous neurodegenerative disorder. Two subtypes commonly present with a language disorder: semantic dementia (SemD) and progressive nonfluent aphasia (PNFA). Patients meeting consensus criteria for PNFA and SemD who had volumetric MRI of sufficient quality to allow cortical thickness analysis were recruited from a tertiary referral clinic: 44 (11 pathologically confirmed) patients with SemD and 32 (4 pathologically confirmed) patients with PNFA and 29 age-matched and gender-matched healthy controls were recruited. Cortical thickness analysis was performed using the Freesurfer software tools. Patients with SemD had significant cortical thinning in the left temporal lobe, particularly temporal pole, entorhinal cortex, and parahippocampal, fusiform, and inferior temporal gyri. A similar but less extensive pattern of loss was seen in the right temporal lobe and (with increasing severity) also in left orbitofrontal, inferior frontal, insular, and cingulate cortices. Patients with PNFA had involvement particularly of the left superior temporal lobe, inferior frontal lobe, and insula, and (with increasing severity) other areas in the left frontal, lateral temporal, and anterior parietal lobes. Similar patterns were seen in the pathologically confirmed cases. Patterns of cortical thinning differed between groups: SemD had significantly more cortical thinning in the temporal lobes bilaterally while PNFA had significantly more thinning in the frontal and parietal lobes. The language variants of frontotemporal lobar degeneration have distinctive and significantly different patterns of cortical thinning. Increasing disease severity is associated with spread of cortical thinning and the pattern of spread is consistent with progression of clinical deficits.


Liquid chromatography/mass spectrometry method for determination of perfluorooctane sulfonyl fluoride upon derivatization with benzylamine.
Perfluorooctane sulfonyl fluoride (PFOSF) is a main precursor of environmentally ubiquitous perfluorooctanesulfonate (PFOS), and the quantity released to the environment is substantial. Determination of PFOSF, particularly at low concentrations, presents significant challenges for high-performance liquid chromatography and liquid chromatography/mass spectrometry (LC/MS) analyses due to the lack of chromophore and ionizable functional group, respectively. In this study, a new method was developed by derivatizing PFOSF with benzylamine to allow rapid quantitative analysis by using LC/MS. The method demonstrated good linearity in the range from 2 to 80 ng mL(-1) with r(2) > 0.994 for the derivatization product while the absolute detection limit was 2.5 pg. Liquid-liquid and liquid-solid extraction procedures were established for analysis of water and soil samples, and recoveries were in the range of 51-128%. In addition, the derivatization was selective for PFOSF, whereas PFOS did not nearly react. The developed simple analytical method with good reproducibility might not only be applied for analysis of PFOSF in the environment but also be applicable for supporting investigations on environmental fate of PFOSF, particularly its environmental and biotransformation to PFOS.


Quantitative evaluation of aerosol deposition pattern in the lung in patients with chronic bronchitis.
A number of numeric measurements have been suggested to describe the pattern of deposition of inhaled aerosols, but there is little information about the relationship between the different measurements. We compared four different methods for analysis of aerosol deposition pattern with respect to their ability to separate patients with chronic bronchitis (CB) from healthy subjects, and with respect to their relationship to a simple index of airflow obstruction. Twelve healthy subjects and 22 patients with CB were studied. All the subjects inhaled a radiolabelled aerosol produced with an air-jet nebulizer. The aerosol contained albumin millimicrospheres labelled with 99Tcm. The ratio of peripheral to central deposition--the penetration index, the coefficient of variation, the coefficient of skewness and the quotient between maximum and mean value deposition were calculated using a gamma camera. The differences between the patients and the healthy subjects were statistically significant for all variables (p < 0.001). The sensitivity of measurements of aerosol deposition for detection of CB was similar to that of forced expiratory volume in one second (FEV1). We found strong correlations between the indices of aerosol deposition and FEV1 between all indices. In clinical routine practice, studies of aerosol deposition offer little advantage in the evaluation of patients with CB. Any of the indices may be suitable for evaluation of aerosol deposition in e.g. studies of inhaled pharmaceuticals.


Sesame Oil Attenuates Renal Oxidative Stress Induced by a High Fat Diet.
The antioxidative effects of the bioactive compounds enriched sesame oil (e.g. lignans and tocopherols) are well established. This study aims to elucidate whether sesame oil could reduce renal oxidative stress induced by a high fat diet (HFD). Mice received HFD for 12 weeks (n=7 per group), which was prepared by adding 20% (w/w) lard (lard group) or sesame oil (sesame group) to the chow diet, respectively. Compared with mice in the lard group, renal lipid levels of those in the sesame group were reduced, shown by decreases in protein expression of transcription factors and enzymes involved in fatty acid synthesis (sterol regulatory element-binding protein-1 and acetyl coenzyme A carboxylase α) and an increase in β-oxidation (peroxisome proliferator-activated receptor α and carnitine palmitoyltransferase I) (P<0.05). In the sesame group, levels of peroxynitrite and thiobarbituric acid reactive substances were also reduced, whereas the level of glutathione was increased. In addition, there was elevated protein expression levels of antioxidant enzymes regulated by nuclear factor-like 2, such as superoxide dismutase, glutathione peroxidase, and glutathione S-transferase (P<0.05), and decreased expression for nuclear factor kappa B and cyclooxygenase 2 (P<0.05). These results suggest that sesame oil could ameliorate HFD-induced renal damage by suppressing oxidative stress and inflammation.


Sequence-specific nuclease-mediated release of cells tethered by oligonucleotide phospholipids.
Single-stranded oligonucleotide-conjugated lipids (ssDNA-PEG-lipids) that associate with the cell membrane confer to the cell an artificial adhesive capability via sequence-specific hybridization to complementary oligonucleotides, forming bonds of double stranded oligonucleotides (dsDNA). Such artificial tethers permit surface patterning of cells or controlled formation of cellular aggregates. However, the hybridization responsible for tethering cells to surfaces or to other cells is not trivially reversed under physiological conditions. In this study, we approach the unbinding of tethered cells by cleaving dsDNA bonds with restriction endonuclease BamHI or digesting bonds with the nonspecific nuclease Benzonase. The procedure was applied to CCRF-CEM cells bearing dsDNA suspended in isolation, cells tethered to glass substrates, and cells aggregated heterotypically with other ssDNA-bearing cells. Cells liberated from surfaces with BamHI could be flushed from flow chambers and viably recovered while the majority of cells not bearing enzyme recognition sequences were retained on the surface, and DNA-tethered cells could be nonspecifically recovered viably from surfaces after Benzonase treatment. Heterotypic aggregates of cells joined by recognition sequence DNA could be dispersed with 10 min exposure to BamHI while undispersed cells heterotypically aggregated with a control sequence remained. Likewise, 10 min exposure to Benzonase was sufficient to disperse aggregates independently of sequence. The potential to undo artificially engineered DNA-mediated adhesion offers new possibilities in the controlled arrangement of cells relative to other cells and in the study of membrane biophysics.


Mandibular alveolar reconstruction using three-dimensional planning.
Resection of malignant disease often results in full-thickness segmental defects of the mandible and loss of dentition. Ideal rehabilitation of the patient requires replacement of the missing segment with vascularized bone that will receive dental implants, and allow osseointegrated prosthetic rehabilitation and restoration of dentition and proper occlusion. Inexact contouring of the bony reconstruction can result in both cosmetic and functional defects that can diminish the future quality of life of the patient. This review summarizes recent advances in preoperative planning and intraoperative techniques that can maximize the success of proper alveolar reconstruction and dental restoration of the patient suffering a segmental maxillary or mandibular defect. Preoperative modeling can be achieved with computer software that utilizes patient imaging. From this imaging, anticipated surgical defects can be planned, models can be generated, and intraoperative templates can be produced. These can be used to improve reconstructive plate bending, relative positioning of bone to opposing jaw, contouring of the reconstructive bone, and even placement of osseointegrated implants in a single operative setting. In patients with complex mandibular defects, the use of computer-assisted three-dimensional planning and modeling can result in time-saving and improved outcomes during maxillary and mandibular reconstruction.


Use of endoscopic ultrasound for diagnosis of cholangiocarcinoma in auto-immune hepatitis.
In this report, a patient was exposed to an herbal remedy for hypercholesterolemia. She became acutely jaundiced while taking the remedy and presented for medical care. Endoscopic ultrasound was utilized, and found a distal common bile duct mass. Endoscopic retrograde cholangiopancreatography guided bile duct biopsies revealed that the mass was cholangiocarcinoma (CCA). This case highlights a unique association between autoimmune hepatitis and CCA. It also highlights that EUS can be safely used in patients with cirrhosis to spare invasive evaluation such as exploratory laporotomy for diagnosis and staging of cholangiocarcinoma.


Sleep and comorbid neurologic disorders.
An understanding of the impact of sleep on neurologic disorders, and the impact of neurologic disorders on sleep, provides fresh opportunities for neurologists to improve the quality of life and functioning of their patients. Sleep-disordered breathing (SDB) is a risk factor for cerebrovascular disease and should be considered in all TIA and stroke patients. Sleep disorders can amplify nociception and worsen headache disorders; and some headaches, including those related to SDB and hypnic headache, are sleep specific. REM sleep behavior disorder may be an early sign of neurodegenerative disease. Focal lesions of almost any etiology (eg, multiple sclerosis and CNS malignancies) in the hypothalamus, basal forebrain, or brainstem may result in sleep disturbance, sleepiness, and insomnia. Sleep-related hypoventilation and fatigue are common in neuromuscular disease. SDB and epilepsy are mutually facilitatory, and poor sleep can exacerbate epilepsy. Continued surveillance for sleep disorders by neurologists is rewarded by new treatment avenues in their patients with the possibility of improved clinical outcomes.


Depressive symptoms in addition to visual impairment, reduced strength and poor balance predict falls in older Taiwanese people.
to determine whether depression is an important and independent predictor of falls in community-dwelling older people living in Taiwan. longitudinal study. five randomly selected villages from Tainan city, Taiwan. in total, 280 community-dwelling people not taking anti-depressant medication aged 65-91 years (mean age 74.9). Participants completed the Geriatric Depression Scale and underwent a range of sensorimotor, balance and mobility tasks and were then followed up for 2 years with monthly telephone calls to determine falls incidence. of the 260 participants with complete follow-up data, 174 (66.9%) experienced no falls, 51 (19.6%) fell once and 35 (13.5%) fell two or more times. Depressive symptoms were significantly more prevalent in recurrent fallers (40.0%) and once-only fallers (27.5%) compared with non-fallers (16.1%). Negative binomial regression analysis identified depression, poor depth perception, reduced lower limb strength and increased sway as independent and significant predictors of falls. depressive symptoms were found to be common in older Taiwanese people and associated with an increased fall risk. These findings suggest that in addition to implementing approaches to maximise vision, strength and balance, fall prevention strategies should also include interventions to assess and treat depression.


The ammonium transporter RhBG: requirement of a tyrosine-based signal and ankyrin-G for basolateral targeting and membrane anchorage in polarized kidney epithelial cells.
RhBG is a nonerythroid member of the Rhesus (Rh) protein family, mainly expressed in the kidney and belonging to the Amt/Mep/Rh superfamily of ammonium transporters. The epithelial expression of renal RhBG is restricted to the basolateral membrane of the connecting tubule and collecting duct cells. We report here that sorting and anchoring of RhBG to the basolateral plasma membrane require a cis-tyrosine-based signal and an association with ankyrin-G, respectively. First, we show by using a model of polarized epithelial Madin-Darby canine kidney cells that the targeting of transfected RhBG depends on a YED motif localized in the cytoplasmic C terminus of the protein. Second, we reveal by yeast two-hybrid analysis a direct interaction between an FLD determinant in the cytoplasmic C-terminal tail of RhBG and the third and fourth repeat domains of ankyrin-G. The biological relevance of this interaction is supported by two observations. (i) RhBG and ankyrin-G were colocalized in vivo in the basolateral domain of epithelial cells from the distal nephron by immunohistochemistry on kidney sections. (ii) The disruption of the FLD-binding motif impaired the membrane expression of RhBG leading to retention on cytoplasmic structures in transfected Madin-Darby canine kidney cells. Mutation of both targeting signal and ankyrin-G-binding site resulted in the same cell surface but nonpolarized expression pattern as observed for the protein mutated on the targeting signal alone, suggesting the existence of a close relationship between sorting and anchoring of RhBG to the basolateral domain of epithelial cells.


The ecological genetics of homoploid hybrid speciation.
Our understanding of homoploid hybrid speciation has advanced substantially since this mechanism of species formation was codified 50 years ago. Early theory and research focused almost exclusively on the importance of chromosomal rearrangements, but it later became evident that natural selection, specifically ecological selection, might play a major role as well. In light of this recent shift, we present an evaluation of ecology's role in homoploid hybrid speciation, with an emphasis on the genetics underlying ecological components of the speciation process. We briefly review new theoretical developments related to the ecology of homoploid hybrid speciation; propose a set of explicit, testable questions that must be answered to verify the role of ecological selection in homoploid hybrid speciation; discuss published work with reference to these questions; and also report new data supporting the importance of ecological selection in the origin of the homoploid hybrid sunflower species Helianthus deserticola. Overall, theory and empirical evidence gathered to date suggest that ecological selection is a major factor promoting homoploid hybrid speciation, with the strongest evidence coming from genetic studies.


Development of Prototype Filovirus Recombinant Antigen Immunoassays.
Throughout the 2014-2015 Ebola outbreak in West Africa, major gaps were exposed in the availability of validated rapid diagnostic platforms, protective vaccines, and effective therapeutic agents. These gaps potentiated the development of prototype rapid lateral flow immunodiagnostic (LFI) assays that are true point-of-contact platforms, for the detection of active Ebola infections in small blood samples. Recombinant Ebola and Marburg virus matrix VP40 and glycoprotein (GP) antigens were used to derive a panel of monoclonal and polyclonal antibodies. Antibodies were tested using a multivariate approach to identify antibody-antigen combinations suitable for enzyme-linked immunosorbent assay (ELISA) and LFI assay development. Polyclonal antibodies generated in goats were superior reagents for capture and detection of recombinant VP40 in test sample matrices. These antibodies were optimized for use in antigen-capture ELISA and LFI assay platforms. Prototype immunoglobulin M (IgM)/immunoglobulin G (IgG) ELISAs were similarly developed that specifically detect Ebola virus-specific antibodies in the serum of experimentally infected nonhuman primates and in blood samples obtained from patients with Ebola from Sierra Leone. The prototype recombinant Ebola LFI assays developed in these studies have sensitivities that are useful for clinical diagnosis of acute ebolavirus infections. The antigen-capture and IgM/IgG ELISAs provide additional confirmatory assay platforms for detecting VP40 and other ebolavirus-specific immunoglobulins.


Microbial Contamination in the Coffee Industry: An Occupational Menace besides a Food Safety Concern?
Respiratory abnormalities among workers at coffee roasting and packaging facilities have already been reported; however, little is known about microbiological contamination inside coffee production facilities. This study intends to assess the microbial contamination (fungi and bacteria) in two coffee industries from Brazil with a multi-approach protocol for sampling and for subsequent analyses using four main sources of samples: filtering respiratory protection devices (FRPD) used by workers, settled dust, electrostatic dust cloths (EDC) and coffee beans. The fungal contamination in the assessed industries was also characterized through the molecular detection of toxigenic species and antifungal resistance. Total bacteria contamination presented the highest values in FRPD collected from both industries (7.45 × 104 CFU·m-2; 1.09 × 104 CFU·m-2). Aspergillus genera was widespread in all the environmental samples collected and sections with clinical relevance (Fumigati) and with toxigenic potential (Nigri and Circumdati) were recovered from FRPD. Circumdati section was observed in 4 mg/mL itraconazole. Sections Circumdati (EDC, coffee beans and settled dust) and Nidulantes (EDC, coffee beans and FRPD) were detected by qPCR. Some of the targeted Aspergillus sections that have been identified microscopically were not detected by qPCR and vice-versa. Overall, this study revealed that microbial contamination is a potential occupational risk in the milling stage and should be tackled when assessing exposure and performing risk assessment. In addition, a multi-sampling campaign should be the approach to follow when assessing microbial contamination and FRPD should be included in this campaign. Occupational exposure to mycotoxins should be considered due to high fungal diversity and contamination. A One Health approach should address these issues in order to prevent consumption of coffee crops and beans infected by fungi and, more specifically, to avoid widespread azole resistance.


The co-occurrence of intimate partner violence and syphilis among pregnant women in Bolivia.
Intimate partner violence (IPV) and sexually transmitted infections (STI) are epidemics that disproportionately affect women. This study determined IPV prevalence and the association between IPV and positive syphilis tests among pregnant women attending antenatal clinics in three provinces in Bolivia. We administered structured questionnaires to women after syphilis testing. The questionnaire included sociodemographic variables and four questions form the modified version of the Abuse Assessment Screen (AAS) to assess physical and sexual violence. Of 6002 women who completed the violence questionnaire and had a syphilis test, 20.4% (n=1227) reported physical or sexual abuse or both committed by their partner in the past year. Prevalence of positive syphilis tests was twice as high among women who reported IPV (8%) than among women who did not (4%) (p<0.01). Women's age (younger), education level (lower), occupation as homemaker, being in a consensual union, more previous pregnancies, lower economic status, and language spoken at home (Spanish and an indigenous language), as well as history of IPV in the past 12 months, were significantly associated with positive syphilis tests in bivariate analysis. History of IPV remained significantly associated with positive syphilis tests in multivariate analysis (OR 1.59, 95% CI 1.23-2.07). In addition, low education among women's partners and having at least one previous pregnancy were positively associated with IPV in multivariate analysis. There is a significant association between history of partner violence and a positive syphilis test among pregnant women, suggesting that syphilis can be an important negative health consequence of IPV. Bolivia's new maternal and infant health program in antenatal clinics, which includes universal syphilis screening, should also provide screening and follow-up care for IPV.


The Columbia University Scale for Psychopathology in Alzheimer's disease.
The Columbia University Scale for Psychopathology in Alzheimer's disease is a new screening instrument developed for use by clinicians or trained lay interviewers. Interrater reliability was established between a psychiatrist and a lay interviewer in 20 patients. In an independent sample of 91 outpatients with very mild to moderate probable Alzheimer's disease, caregiver informants reported that depressed mood was common (46.2%) but rarely persistent (2.2%), and that sleep disturbance occurred frequently (41.8%) but was never severe (0%). There were significant but weak associations between the presence of specific subtypes of delusions and severity of dementia. Although a variety of delusional symptoms were reported, they were frequently transient and patients often accepted the truth if corrected by the caregiver. As a result, few patients met broad or narrow operational criteria used to define delusions. Prior studies may have overestimated the prevalence of psychotic features in Alzheimer's disease by not employing standard definitional criteria. The findings also indicate that new methodology such as that employed in this instrument needs to be evaluated more widely.


Liver tumours in children: Current surgical management and role of Transplantation.
This article reviews the current surgical management of liver tumours in children in the light of improved chemotherapy, surgical techniques and outcomes from transplantation. It is a principle of management that complete removal of a tumour must be achieved for cure. Neoadjuvant chemotherapy may downstage advanced local disease to enable safe curative tumour resection. When this is not achievable, transplant is indicated. Conventional indications for transplant are unresectable stages 3 and 4 tumours confined to the liver. With the realisation that lifelong immunosuppressive therapy has considerable adverse consequences, there has been a recent trend towards extreme and 'acrobatic' liver resection to avoid transplantation, but still obtain a cure. The current literature is reviewed in the light of these trends and our own experience.


Myocarditis-inducing epitope of myosin binds constitutively and stably to I-Ak on antigen-presenting cells in the heart.
Immune interactions in the heart were studied using a murine model of myosin-induced autoimmune myocarditis. A T cell hybridoma specific for mouse cardiac myosin was generated from A/J mice and used to demonstrate that endogenous myosin/I-Ak complexes are constitutively expressed on antigen-presenting cells in the heart. This T cell hybridoma, Seu.5, was used as a functional probe to identify a myocarditis-inducing epitope of cardiac myosin. Overlapping peptides based on the cardiac myosin heavy chain alpha (myhc alpha) sequences were synthesized and tested for their ability to stimulate Seu.5 T cells. One peptide, myhc alpha (325-357) strongly stimulated the Seu.5 T cells, localizing the epitope to this region of the myhc alpha molecule. Using truncated peptides, the epitope was further localized to residues 334-352. The myhc alpha (334-352) peptide strongly induced myocarditis when administered to A/J mice, which was histologically indistinguishable from that induced by myosin. The myhc alpha (334-352) epitope was present in cardiac myosin and not skeletal muscle myosins, providing a biochemical basis for the cardiac specificity of this autoimmune disease. Induction of myocarditis by this epitope was restricted to the myhc alpha isoform and not the myhc beta isoform, suggesting there may be a difference in the efficiency of generating tolerance to these isoforms of cardiac myosin, which are differentially developmentally regulated. The myhc alpha (334-352) epitope bound to purified I-Ak molecules in a similar manner to other I-Ak-restricted immunogenic epitopes, HEL(48-61) and RNase(43-56). Importantly, the myhc alpha (334-352) epitope was able to bind to I-Ak molecules on the surface of antigen-presenting cells in a stable manner. These findings demonstrate that autoantigenic epitopes can behave in a dominant manner and constitutively bind to class II molecules in the target organ in a similar manner to foreign immunogenic epitopes.


POEMS syndrome demonstrating VEGF decrease by ticlopidine.
POEMS syndrome is a syndrome that presents with polyneuropathy, organomegaly, endocrinopathy, M-proteins and skin changes. Elevated vascular endothelial growth factor (VEGF) has recently been reported in POEMS syndrome. We report a case of POEMS syndrome with high VEGF titers. Steroid, plasmapheresis and intravenous gamma-globulin had little effect. Various immunosuppressive agents were discontinued due to side effects. Although administration of aspirin did not decrease VEGF, ticlopidine decreased VEGF significantly. This case suggests that ticlopidine is a candidate for supportive therapy in POEMS syndrome and we should measure VEGF before and after the administration of ticlopidine in other cases.