Targeted next generation sequencing of the entire vitamin D receptor gene reveals polymorphisms correlated with vitamin D deficiency among older Filipino women with and without fragility fracture.
This study aimed to discover genetic variants in the entire 101 kB vitamin D receptor (VDR) gene for vitamin D deficiency in a group of postmenopausal Filipino women using targeted next generation sequencing (TNGS) approach in a case-control study design. A total of 50 women with and without osteoporotic fracture seen at the Philippine Orthopedic Center were included. Blood samples were collected for determination of serum vitamin D, calcium, phosphorus, glucose, blood urea nitrogen, creatinine, aspartate aminotransferase, alanine aminotransferase and as primary source for targeted VDR gene sequencing using the Ion Torrent Personal Genome Machine. The variant calling was based on the GATK best practice workflow and annotated using Annovar tool. A total of 1496 unique variants in the whole 101-kb VDR gene were identified. Novel sequence variations not registered in the dbSNP database were found among cases and controls at a rate of 23.1% and 16.6% of total discovered variants, respectively. One disease-associated enhancer showed statistically significant association to low serum 25-hydroxy vitamin D levels (Pearson chi-square P-value=0.009). The transcription factor binding site prediction program PROMO predicted the disruption of three transcription factor binding sites in this enhancer region. These findings show the power of TNGS in identifying sequence variations in a very large gene and the surprising results obtained in this study greatly expand the catalog of known VDR sequence variants that may represent an important clue in the emergence of vitamin D deficiency. Such information will also provide the additional guidance necessary toward a personalized nutritional advice to reach sufficient vitamin D status.


Endovascular treatment of intracranial aneurysms: a minimally invasive approach with advantages for elderly patients.
An endovascular approach, embolizing intracranial aneurysms with electrolytically detachable coils, is a new minimally invasive treatment of aneurysms. Reviewing our experience with 13 patients aged 70 years or more, 12 patients were treated successfully. Over an average follow-up period of 8 months, there was only one death which might be attributed to an intracranial aneurysm. This compares favourably with mortality rates approaching 50% for either conservative or traditional surgical management of aneurysms presenting with subarachnoid haemorrhage in this age group. Coil embolization, avoiding craniotomy, may offer particular advantages in managing intracranial aneurysms in the elderly patient.


Mitochondrial DNA level, but not active replicase, is essential for Caenorhabditis elegans development.
A number of studies showed that the development and the lifespan of Caenorhabditis elegans is dependent on mitochondrial function. In this study, we addressed the role of mitochondrial DNA levels and mtDNA maintenance in development of C. elegans by analyzing deletion mutants for mitochondrial polymerase gamma (polg-1(ok1548)). Surprisingly, even though previous studies in other model organisms showed necessity of polymerase gamma for embryonic development, homozygous polg-1(ok1548) mutants had normal development and reached adulthood without any morphological defects. However, polg-1 deficient animals have a seriously compromised gonadal function as a result of severe mitochondrial depletion, leading to sterility and shortened lifespan. Our results indicate that the gonad is the primary site of mtDNA replication, whilst the mtDNA of adult somatic tissues mainly stems from the developing embryo. Furthermore, we show that the mtDNA copy number shows great plasticity as it can be almost tripled as a response to the environmental stimuli. Finally, we show that the mtDNA copy number is an essential limiting factor for the worm development and therefore, a number of mechanisms set to maintain mtDNA levels exist, ensuring a normal development of C. elegans even in the absence of the mitochondrial replicase.


Rivaroxaban versus warfarin in nonvalvular atrial fibrillation.
The use of warfarin reduces the rate of ischemic stroke in patients with atrial fibrillation but requires frequent monitoring and dose adjustment. Rivaroxaban, an oral factor Xa inhibitor, may provide more consistent and predictable anticoagulation than warfarin. In a double-blind trial, we randomly assigned 14,264 patients with nonvalvular atrial fibrillation who were at increased risk for stroke to receive either rivaroxaban (at a daily dose of 20 mg) or dose-adjusted warfarin. The per-protocol, as-treated primary analysis was designed to determine whether rivaroxaban was noninferior to warfarin for the primary end point of stroke or systemic embolism. In the primary analysis, the primary end point occurred in 188 patients in the rivaroxaban group (1.7% per year) and in 241 in the warfarin group (2.2% per year) (hazard ratio in the rivaroxaban group, 0.79; 95% confidence interval [CI], 0.66 to 0.96; P<0.001 for noninferiority). In the intention-to-treat analysis, the primary end point occurred in 269 patients in the rivaroxaban group (2.1% per year) and in 306 patients in the warfarin group (2.4% per year) (hazard ratio, 0.88; 95% CI, 0.74 to 1.03; P<0.001 for noninferiority; P=0.12 for superiority). Major and nonmajor clinically relevant bleeding occurred in 1475 patients in the rivaroxaban group (14.9% per year) and in 1449 in the warfarin group (14.5% per year) (hazard ratio, 1.03; 95% CI, 0.96 to 1.11; P=0.44), with significant reductions in intracranial hemorrhage (0.5% vs. 0.7%, P=0.02) and fatal bleeding (0.2% vs. 0.5%, P=0.003) in the rivaroxaban group. In patients with atrial fibrillation, rivaroxaban was noninferior to warfarin for the prevention of stroke or systemic embolism. There was no significant between-group difference in the risk of major bleeding, although intracranial and fatal bleeding occurred less frequently in the rivaroxaban group. (Funded by Johnson & Johnson and Bayer; ROCKET AF ClinicalTrials.gov number, NCT00403767.).


Classifying murine glomerulonephritis using optical coherence tomography and optical coherence elastography.
Acute glomerulonephritis caused by antiglomerular basement membrane marked by high mortality. The primary reason for this is delayed diagnosis via blood examination, urine analysis, tissue biopsy, or ultrasound and X-ray computed tomography imaging. Blood, urine, and tissue-based diagnoses can be time consuming, while ultrasound and CT imaging have relatively low spatial resolution, with reduced sensitivity. Optical coherence tomography is a noninvasive and high-resolution imaging technique that provides superior spatial resolution (micrometer scale) as compared to ultrasound and CT. Changes in tissue properties can be detected based on the optical metrics analyzed from the OCT signals, such as optical attenuation and speckle variance. Furthermore, OCT does not rely on ionizing radiation as with CT imaging. In addition to structural changes, the elasticity of the kidney can significantly change due to nephritis. In this work, OCT has been utilized to quantify the difference in tissue properties between healthy and nephritic murine kidneys. Although OCT imaging could identify the diseased tissue, its classification accuracy is clinically inadequate. By combining optical metrics with elasticity, the classification accuracy improves from 76% to 95%. These results show that OCT combined with OCE can be a powerful tool for identifying and classifying nephritis. Therefore, the OCT/OCE method could potentially be used as a minimally invasive tool for longitudinal studies during the progression and therapy of glomerulonephritis as well as complement and, perhaps, substitute highly invasive tissue biopsies. Elastic-wave propagation in mouse healthy and nephritic kidneys.


Biallelic SQSTM1 mutations in early-onset, variably progressive neurodegeneration.
To characterize clinically and molecularly an early-onset, variably progressive neurodegenerative disorder characterized by a cerebellar syndrome with severe ataxia, gaze palsy, dyskinesia, dystonia, and cognitive decline affecting 11 individuals from 3 consanguineous families. We used whole-exome sequencing (WES) (families 1 and 2) and a combined approach based on homozygosity mapping and WES (family 3). We performed in vitro studies to explore the effect of the nontruncating SQSTM1 mutation on protein function and the effect of impaired SQSTM1 function on autophagy. We analyzed the consequences of sqstm1 down-modulation on the structural integrity of the cerebellum in vivo using zebrafish as a model. We identified 3 homozygous inactivating variants, including a splice site substitution (c.301+2T>A) causing aberrant transcript processing and accelerated degradation of a resulting protein lacking exon 2, as well as 2 truncating changes (c.875_876insT and c.934_936delinsTGA). We show that loss of SQSTM1 causes impaired production of ubiquitin-positive protein aggregates in response to misfolded protein stress and decelerated autophagic flux. The consequences of sqstm1 down-modulation on the structural integrity of the cerebellum in zebrafish documented a variable but reproducible phenotype characterized by cerebellum anomalies ranging from depletion of axonal connections to complete atrophy. We provide a detailed clinical characterization of the disorder; the natural history is reported for 2 siblings who have been followed up for >20 years. This study offers an accurate clinical characterization of this recently recognized neurodegenerative disorder caused by biallelic inactivating mutations in SQSTM1 and links this phenotype to defective selective autophagy.


Hyperhomocyst(e)inemia, related vitamins and dementias.
Vitamin B12 and to a lesser extent folate deficiencies have been associated with dementias. Both these vitamins are determinants of plasma total homocysteine concentrations. In this review the frequency distributions of plasma vitamin B12, folate and homocysteine in South African males (# 51 yrs and > 51 yrs) illustrate the lower vitamin B12 levels in older subjects, and the shift toward elevated homocysteine concentrations in elderly people. Vitamin B12 deficiency appears to be associated with neuropsychiatric disorders, including dementias, but no causal relationship based on biochemical evidence has so far been established. Supplementation with vitamin B12 improves some neurological abnormalities and reverses only mild dementia of recent onset, but does not slow the progression of dementia. Elevated homocysteine levels appears to affect cognitive function, as measured by spatial copying skills and visual event-related potentials. Measurement of plasma homocysteine may help identify individuals with vitamin deficiencies and hyperhomocysteinemia. The relation between B-vitamins, homocysteine and dementia needs to explored further before vitamin supplementation is advocated to prevent or reverse neuropsychiatric disorders.


An integrated management strategy to prevent outbreaks and eliminate infection pressure of American foulbrood disease in a commercial beekeeping operation.
The bacterial disease American Foulbrood (AFB), caused by the Gram-positive bacterium Paenibacillus larvae, is considered the most contagious and destructive infectious disease affecting honeybees world-wide. The resilient nature of P. larvae bacterial spores presents a difficult problem for the control of AFB. Burning clinically symptomatic colonies is widely considered the only workable strategy to prevent further spread of the disease. Antibiotic use is banned in EU countries, and although used commonly in the U.S. and Canada, it only masks symptoms and does not prevent the further spread of the disease. Not surprisingly, there is an increased demand for chemical-free strategies to prevent and control of AFB. The aim of this study was to implement a management program with a long-term perspective to reduce infection pressure and eliminate AFB outbreaks. The study was conducted within a commercial beekeeping operation in central Sweden that has previously experienced reoccurring AFB outbreaks. For 5 years, P. larvae were cultured from adult bee samples taken in the fall. The following spring, any identified sub-clinically infected colonies were shaken onto new material and quarantined from the rest of the beekeeping operation. After the first year clinical symptoms were not again observed, and during the 5 years of the study the proportion of apiaries harbouring P. larvae spores decreased from 74% to 4%. A multinomial regression analysis also clearly demonstrated that the proportion of infected colonies with the highest levels of spore counts disproportionately declined so that by the end of the study the only remaining infected apiaries were in the lowest spore count category (the three higher spore count categories having been eradicated). These results demonstrate the importance of management practices on AFB disease epidemiology. Early detection of subclinical spore prevelance and quarantine management as presented here can provide an effective sustainable chemical-free preventive solution to reduce both the incidence of AFB outbreaks and continued transmission risk at a large-scale.


Epidemiology of bacterial meningitis in the USA from 1997 to 2010: a population-based observational study.
Bacterial meningitis continues to be a substantial cause of morbidity and mortality, but the epidemiological trends after adjunctive dexamethasone recommendations are unknown in the USA. We aimed to describe the changing patterns among the most common bacterial causes in the USA after conjugate vaccination and to assess the association between adjunctive dexamethasone and mortality. For this population-based observational study, we searched information available from hospital discharges about incidence and inpatient mortality for the most important causes of community and nosocomial bacterial meningitis based on International Classification of Diseases coding across all hospitals in the USA between 1997 and 2010 with the HealthCare Cost Utilization Project (HCUP) network database. We calculated incidences according to US Census Bureau data and used a negative binomial regression model to evaluate the significance of changes over time. We assessed mortality from pneumococcus for three periods 1997-2001 (baseline), 2002-04 (transition years), and 2005-08 (after corticosteroid recommendations were available). Streptococcus pneumoniae incidence fell from 0·8 per 100 000 people in 1997, to 0·3 per 100 000 people by the end of 2010 (RR 0·3737, 95% CI 0·1825-0·7656). Mortality from pneumococcal meningitis decreased between 2005 (0·049 per 100 000 people) and 2008 (0·024 per 100 000 people) compared with between 2002 (0·073 per 100 000 people) and 2004 (0·063 per 100 000 people; RR 0·5720, 95% CI 0·4303-0·7582). The incidence of Neisseria meningitidis infection decreased from 0·721 per 100 000 people in 1997, to 0·123 per 100 000 people in 2010 (RR 0·1386, 95% CI 0·048-0·4284), which has placed this pathogen close to common bacterial causes of nosocomial meningitis such as staphylococcus and Gram-negative bacteria and to Haemophilus influenzae. S pneumoniae continues to be the leading identifiable cause of bacterial meningitis in the USA, but with a significant decrease in incidence and mortality associated with the introduction of conjugated vaccines and a mortality decrease that is associated with the introduction of recommendations for use of adjunctive dexamethasone for pneumococcal meningitis. National Center for Research Resources.


Polyimide and SU-8 microfluidic devices manufactured by heat-depolymerizable sacrificial material technique.
The following paper describes a sacrificial layer method for the manufacturing of microfluidic devices in polyimide and SU-8. The technique uses heat-depolymerizable polycarbonates embedded in polyimide or SU-8 for the generation of microchannels and sealed cavities. The volatile decomposition products originating from thermolysis of the sacrificial material escape out of the embedding material by diffusion through the cover layer. The fabrication process was studied experimentally and theoretically with a focus on the decomposition of the sacrificial materials and their diffusion through the polyimide or SU-8 cover layer. It is demonstrated that the sacrificial material removal process is independent of the actual channel geometry and advances linearly with time unlike conventional sacrificial layer techniques. The fabrication method provides a versatile and fast technique for the manufacturing of microfluidic devices for applications in the field of microTAS and Lab-on-a-Chip.


Developmental control of a G1-S transcriptional program in Drosophila.
We have defined a coordinate program of transcription of S-phase genes (DNA polymerase alpha, PCNA and the two ribonucleotide reductase subunits) that can be induced by the G1 cyclin, cyclin E. In Drosophila embryos, this program drives an intricate spatial and temporal pattern of gene expression that perfectly parallels the embryonic program of S-phase control. This dynamic pattern of expression is not disrupted by a mutation, string, that blocks the cell cycle. Thus, the transcriptional program is not a secondary consequence of cell cycle progression. We suggest that developmental signals control this transcriptional program and that its activation either directly or indirectly drives transition from G1 to S phase in the stereotyped embryonic pattern.


Management of gallbladder polyps.
Polyps of the gallbladder are often incidentally identified during radiographic evaluation of abdominal pain. Once identified, they pose a dilemma with respect to their proper long-term management. Herein, polyps of the gallbladder are reviewed with specific attention to their management in light of their malignant potential. The available data suggest that polyps larger than 10 mm or that show rapid growth should be considered suggestive of malignancy, prompting cholecystectomy.


Elucidation of salicylate attachment in celesticetin biosynthesis opens the door to create a library of more efficient hybrid lincosamide antibiotics.
Lincosamides, which are produced by streptomycetes, compose a small but clinically important class of antibiotics. The recent elucidation of the condensation and post-condensation biosynthetic steps of the lincosamides lincomycin and celesticetin revealed several unexpected reaction mechanisms. Here, we prepared recombinant proteins involved in the celesticetin biosynthetic pathway and used them for in vitro assays that were monitored by LC-MS. Our results elucidate the last biosynthetic step of celesticetin: the attachment of salicylic acid is catalyzed by the Ccb2 acyl-CoA ligase and the Ccb1 acyltransferase. Ccb1 belongs to the WS/DGAT protein family and, in contrast to the characterized members of the family, has unusual substrate specificity. To the best of our knowledge, Ccb1 is the first protein in this family that transfers a benzoyl derivative-CoA conjugate and is the first WS/DGAT protein involved in the biosynthesis of secondary metabolites. Furthermore, we exploited the relaxed substrate specificities of Ccb1 and Ccb2, as well as three additional upstream post-condensation biosynthetic proteins in the celesticetin pathway, and combined the lincomycin and the celesticetin biosynthetic pathways in vitro. In this way, we prepared a library of 150 novel hybrid lincosamides, including two unnatural chimeras of lincomycin and celesticetin, which were shown to have antibacterial properties more pronounced than clinically used lincomycin. These achievements may be considered a case study in applying knowledge about biosynthetic machinery to assemble a large number of compounds from originally a small group of natural products without the need for chemical synthesis.


Prevalence, predictors and outcome of hypofibrinogenaemia in trauma: a multicentre observational study.
Exsanguination due to trauma-induced coagulopathy is a continuing challenge in emergency trauma care. Fibrinogen is a crucial factor for haemostatic competence, and may be the factor that reaches critically low levels first. Early fibrinogen substitution is advocated by a number of authors. Little evidence exists regarding the indications for fibrinogen supplementation in the acute phase. This study aims to estimate the prevalence of hypofibrinogenaemia in a multi-center trauma population, and to explore how initial fibrinogen concentration relates to outcome. Also, factors contributing to low fibrinogen levels are identified. Patients arriving in hospital less than 180 minutes post-injury requiring full trauma team activation in four different centers were included in the study. Time from injury, patient demographics, injury severity scores (ISS) and 28 days outcome status were recorded. Initial blood samples for coagulation and blood gas were analyzed. Generalized additive regression, piecewise linear regression, and multiple linear regression models were used for data analyses. Out of 1,133 patients we identified a fibrinogen concentration ≤1.5g/L in 8.2%, and <2 g/L in 19.2%. A non-linear relationship between fibrinogen concentration and mortality was detected in the generalized additive and piecewise linear regression models. In the piecewise linear regression model we identified a breakpoint for optimal fibrinogen concentration at 2.29 g/L (95% confidence interval (CI): 1.93 to 2.64). Below this value the odds of death by 28 days was reduced by a factor of 0.08 (95% CI: 0.03 to 0.20) for every unit increase in fibrinogen concentration. Low age, male gender, lengthened time from injury, low base excess and high ISS were unique contributors to low fibrinogen concentrations on arrival. Hypofibrinogenaemia is common in trauma and strongly associated with poor outcome. Below an estimated critical fibrinogen concentration value of 2.29 g/L a dramatic increase in mortality was detected. This finding indicates that the negative impact of low fibrinogen concentrations may have been previously underestimated. A number of clinically identifiable factors are associated with hypofibrinogenaemia. They should be considered in the management of massively bleeding patients. Interventional trials with fibrinogen substitution in high-risk patients need to be undertaken.


Virulence in malaria: an evolutionary viewpoint.
Malaria parasites cause much morbidity and mortality to their human hosts. From our evolutionary perspective, this is because virulence is positively associated with parasite transmission rate. Natural selection therefore drives virulence upwards, but only to the point where the cost to transmission caused by host death begins to outweigh the transmission benefits. In this review, we summarize data from the laboratory rodent malaria model, Plasmodium chabaudi, and field data on the human malaria parasite, P. falciparum, in relation to this virulence trade-off hypothesis. The data from both species show strong positive correlations between asexual multiplication, transmission rate, infection length, morbidity and mortality, and therefore support the underlying assumptions of the hypothesis. Moreover, the P. falciparum data show that expected total lifetime transmission of the parasite is maximized in young children in whom the fitness cost of host mortality balances the fitness benefits of higher transmission rates and slower clearance rates, thus exhibiting the hypothesized virulence trade-off. This evolutionary explanation of virulence appears to accord well with the clinical and molecular explanations of pathogenesis that involve cytoadherence, red cell invasion and immune evasion, although direct evidence of the fitness advantages of these mechanisms is scarce. One implication of this evolutionary view of virulence is that parasite populations are expected to evolve new levels of virulence in response to medical interventions such as vaccines and drugs.


Prevalence of exercise-induced bronchoconstriction and exercise-induced laryngeal obstruction in a general adolescent population.
Exercise-induced respiratory symptoms are common among adolescents. Exercise is a known stimulus for transient narrowing of the airways, such as exercise-induced bronchoconstriction (EIB) and exercise-induced laryngeal obstruction (EILO). Our aim was to investigate the prevalence of EIB and EILO in a general population of adolescents. In this cross-sectional study, a questionnaire on exercise-induced dyspnoea was sent to all adolescents born in 1997 and 1998 in Uppsala, Sweden (n=3838). A random subsample of 146 adolescents (99 with self-reported exercise-induced dyspnoea and 47 without this condition) underwent standardised treadmill exercise tests for EIB and EILO. The exercise test for EIB was performed while breathing dry air; a positive test was defined as a decrease of ≥10% in FEV1 from baseline. EILO was investigated using continuous laryngoscopy during exercise. The estimated prevalence of EIB and EILO in the total population was 19.2% and 5.7%, respectively. No gender differences were found. In adolescents with exercise-induced dyspnoea, 39.8% had EIB, 6% had EILO and 4.8% had both conditions. In this group, significantly more boys than girls had neither EIB nor EILO (64.7% vs 38.8%; p=0.026). There were no significant differences in body mass index, lung function, diagnosed asthma or medication between the participants with exercise-induced dyspnoea who had or did not have a positive EIB or EILO test result. Both EIB and EILO are common causes of exercise-induced dyspnoea in adolescents. EILO is equally common among girls and boys and can coexist with EIB.


Small supernumerary marker chromosomes: A legacy of trisomy rescue?
We studied by a whole genomic approach and trios genotyping, 12 de novo, nonrecurrent small supernumerary marker chromosomes (sSMC), detected as mosaics during pre- or postnatal diagnosis and associated with increased maternal age. Four sSMCs contained pericentromeric portions only, whereas eight had additional non-contiguous portions of the same chromosome, assembled together in a disordered fashion by repair-based mechanisms in a chromothriptic event. Maternal hetero/isodisomy was detected with a paternal origin of the sSMC in some cases, whereas in others two maternal alleles in the sSMC region and biparental haplotypes of the homologs were detected. In other cases, the homologs were biparental while the sSMC had the same haplotype of the maternally inherited chromosome. These findings strongly suggest that most sSMCs are the result of a multiple-step mechanism, initiated by maternal meiotic nondisjunction followed by postzygotic anaphase lagging of the supernumerary chromosome and its subsequent chromothripsis.


T-cadherin is critical for adiponectin-mediated cardioprotection in mice.
The circulating, adipocyte-secreted hormone adiponectin (APN) exerts protective effects on the heart under stress conditions. The receptors binding APN to cardiac tissue, however, have remained elusive. Here, we report that the glycosyl phosphatidylinositol–anchored cell surface glycoprotein T-cadherin (encoded by Cdh13) protects against cardiac stress through its association with APN in mice. We observed extensive colocalization of T-cadherin and APN on cardiomyocytes in vivo. In T-cadherin-deficient mice, APN failed to associate with cardiac tissue, and its levels dramatically increased in the circulation. Pressure overload stress resulted in exacerbated cardiac hypertrophy in T-cadherin-null mice and paralleled corresponding defects in mice lacking APN. During ischemia-reperfusion injury, the absence of T-cadherin increased infarct size similar to that in APN-null mice. Myocardial AMPK is a major downstream protective signaling target of APN. In both cardiac hypertrophy and ischemia-reperfusion models, T-cadherin was necessary for APN-dependent AMPK phosphorylation. In APN-null mice, recombinant adenovirus-expressed APN reduced exaggerated hypertrophy and infarct size and restored AMPK phosphorylation as previously reported. In contrast, rescue was ineffective in mice lacking T-cadherin in addition to APN. These data suggest that T-cadherin protects from stress-induced pathological cardiac remodeling by binding APN and activating its cardioprotective functions.


Comparison of adherence rates for antiretroviral, blood pressure, or mental health medications for HIV-positive patients at an academic medical center outpatient pharmacy.
Despite advances in safety, tolerability, and decreased pill burden for human immunodeficiency virus (HIV) antiretroviral therapy (ART), nonadherence continues to be a major cause of HIV-related morbidity and mortality. Adherence to these medications presents particular challenges because persons living with HIV (PLWH) must use medication more consistently (≥ 95%) than persons with other chronic diseases, such as hypertension or diabetes mellitus (≥ 80%). To (a) quantify PLWH adherence rates to scheduled ART, blood pressure medications, or mental health medications, (b) examine how pill burden and dosing schedule affected adherence, and (c) analyze a non-HIV comparator group for comparison with PLWH patients on adherence rates. Patients ≥ 18 years of age, on any prescribed HIV ART, scheduled prescription blood pressure medication, or scheduled prescription mental health medication filled between March 1, 2012, and March 31, 2013, were included in the analysis of data from the University of Colorado Hospital outpatient pharmacy system. The proportion of days covered (PDC) was calculated to measure adherence. Statistical analyses were performed to compare adherence rates between groups. The study included 865 PLWH, who filled 1,943 antiviral prescriptions with an average PDC of 84.5%. However, only 40% of patients had high enough adherence to achieve therapeutic benefits based on a criterion of at least 95% of scheduled doses taken. When separated by regimen, 282 patients filled single-tablet once-daily regimens; 295 patients filled multitablet once-daily regimens; and 288 patients filled multitablet twice-daily regimens. The calculated PDC of PLWH on a single-tablet once-daily regimen was 89.7% versus 81.0% for PLWH on a multitablet once-daily regimen (P less than 0.001). The average PDC for PLWH (n = 269) who filled 460 scheduled blood pressure prescriptions was 82.7%. The average PDC for PLWH (n = 295) filling 467 scheduled mental health prescriptions was 81.7%. This difference was statistically significant in adherence between ART and blood pressure medications of 2.5% (P = 0.013) and a difference between ART and scheduled mental health medications of 3.03% (P = 0.002). For the comparator group of similar patients without HIV, 895 patients filled 1,398 scheduled blood pressure medications, with an average adherence of 83.8%. The difference between groups was not statistically significant (P = 0.390). For scheduled mental health medications in the comparator group, there were 666 patients filling 828 prescriptions. The comparator group had a higher average PDC than PLWH, with 85.1% versus 81.7%, respectively (P = 0.009). Based on average PDC, PLWH filling prescriptions at the University of Colorado Hospital Infectious Disease Group Practice pharmacy had an adherence of 84.5% to ART. However, only 40% of patients were adherent at the needed 95% level for therapeutic effects. Additionally, PLWH were more adherent to single-tablet once-daily regimens than to multitablet once-daily regimens or multitablet twice-daily regimens. Adherence in PLWH to HIV ART was better than the same patients' adherence to scheduled blood pressure and scheduled mental health medications. Levels of adherence still need to be improved to optimal to reduce rates of resistance and maximize therapeutic durability of selected regimens. When analyzed with the comparator group, adherence in PLWH to scheduled blood pressure medications was similar, but adherence to scheduled mental health medications was slightly worse. Further work is needed to address nonadherence among PLWH, including their adherence both to ART and to medications prescribed for other chronic diseases.


A protocol for ultrastructural study of Candida albicans biofilm using transmission electron microscopy.
This protocol describes how to analyze C. albicans biofilm using transmission electron microscopy. We present two approaches to observe the ultrastructure of fungal cells within unperturbed biofilms, as well as an immunogold labeling procedure. This approach maintains the architecture of the fungal biofilm close to its native state by growing C. albicans biofilm on a plastic surface. After the freeze substitution procedure, classical transmission electron microscopy or electron tomography will allow the ultrastructural analysis of the microbial community.


Nitric oxide-dependent activation of CaMKII increases diastolic sarcoplasmic reticulum calcium release in cardiac myocytes in response to adrenergic stimulation.
Spontaneous calcium waves in cardiac myocytes are caused by diastolic sarcoplasmic reticulum release (SR Ca(2+) leak) through ryanodine receptors. Beta-adrenergic (β-AR) tone is known to increase this leak through the activation of Ca-calmodulin-dependent protein kinase (CaMKII) and the subsequent phosphorylation of the ryanodine receptor. When β-AR drive is chronic, as observed in heart failure, this CaMKII-dependent effect is exaggerated and becomes potentially arrhythmogenic. Recent evidence has indicated that CaMKII activation can be regulated by cellular oxidizing agents, such as reactive oxygen species. Here, we investigate how the cellular second messenger, nitric oxide, mediates CaMKII activity downstream of the adrenergic signaling cascade and promotes the generation of arrhythmogenic spontaneous Ca(2+) waves in intact cardiomyocytes. Both SCaWs and SR Ca(2+) leak were measured in intact rabbit and mouse ventricular myocytes loaded with the Ca-dependent fluorescent dye, fluo-4. CaMKII activity in vitro and immunoblotting for phosphorylated residues on CaMKII, nitric oxide synthase, and Akt were measured to confirm activity of these enzymes as part of the adrenergic cascade. We demonstrate that stimulation of the β-AR pathway by isoproterenol increased the CaMKII-dependent SR Ca(2+) leak. This increased leak was prevented by inhibition of nitric oxide synthase 1 but not nitric oxide synthase 3. In ventricular myocytes isolated from wild-type mice, isoproterenol stimulation also increased the CaMKII-dependent leak. Critically, in myocytes isolated from nitric oxide synthase 1 knock-out mice this effect is ablated. We show that isoproterenol stimulation leads to an increase in nitric oxide production, and nitric oxide alone is sufficient to activate CaMKII and increase SR Ca(2+) leak. Mechanistically, our data links Akt to nitric oxide synthase 1 activation downstream of β-AR stimulation. Collectively, this evidence supports the hypothesis that CaMKII is regulated by nitric oxide as part of the adrenergic cascade leading to arrhythmogenesis.


NK-cell neoplasms in Japan.
Neoplasms putatively originating from precursor and mature natural killer (NK) cells are rare, and their clinical features are unclear. A nationwide survey was performed in Japan to clarify the clinical features of these neoplasms diagnosed between 1994 and 1998, and data for 237 patients who met the criteria for putative NK cell-lineage neoplasms were analyzed. Among them, 11 had myeloid/NK-cell precursor acute leukemia, 15 blastic NK-cell lymphoma, 21 precursor NK-cell acute lymphoblastic leukemia, 22 aggressive NK-cell leukemia/lymphoma, 149 nasal-type NK-cell lymphoma (123 nasal and 26 extranasal) and 19 chronic NK lymphocytosis. The median overall survival time of patients with aggressive NK-cell leukemia/lymphoma was 2 months, which for chronic NK lymphocytosis was more than 8 years, and that for the other types of NK-cell neoplasms was between 6 and 22 months. Nasal NK-cell lymphoma and extranasal NK-cell lymphoma share the same histology. The age of affliction was the same, but the sex was different with males predominantly having nasal NK-cell lymphoma and females extranasal NK-cell lymphoma. Patients with extranasal NK-cell lymphoma had the tendency to exhibit a more advanced state of disease, with significantly higher International Prognostic Index and LDH levels, and significantly lower hemoglobin and platelet levels. The overall survival, however, did not differ significantly. Precursor NK-cell acute lymphoblastic leukemia and blastic NK-cell lymphoma were arbitrarily defined by the presence or absence of 30% or more of blastic cells in the bone marrow or peripheral blood, but there were no significant differences for affected age, gender, involved sites or prognosis. Aggressive NK-cell leukemia/lymphoma and extranasal NK-cell lymphoma were arbitrarily defined by the presence or absence of 30% or more of large granular lymphocytes in the bone marrow or peripheral blood and it is possible that aggressive NK-cell leukemia/lymphoma is a leukemic phase of extranasal NK-cell lymphoma. The incidence of skin involvement, however, was significantly higher for extranasal NK-cell lymphoma, suggesting that the two diseases are different. In nasal NK-cell lymphoma, Epstein-Barr virus in tumor cells was detected in all patients tested, suggesting its causative role.


Sanfilippo syndrome: a mini-review.
Mucopolysaccharidosis type III (MPS III, Sanfilippo syndrome) is an autosomal recessive disorder, caused by a deficiency in one of the four enzymes involved in the lysosomal degradation of the glycosaminoglycan heparan sulfate. Based on the enzyme deficiency, four different subtypes, MPS IIIA, B, C, and D, are recognized. The genes encoding these four enzymes have been characterized and various mutations have been reported. The probable diagnosis of all MPS III subtypes is based on increased concentration of heparan sulfate in the urine. Enzymatic assays in leukocytes and/or fibroblasts confirm the diagnosis and allow for discrimination between the different subtypes of the disease. The clinical course of MPS III can be divided into three phases. In the first phase, which usually starts between 1 and 4 years of age, a developmental delay becomes apparent after an initial normal development during the first 1-2 years of life. The second phase generally starts around 3-4 years and is characterized by severe behavioural problems and progressive mental deterioration ultimately leading to severe dementia. In the third and final stage, behavioural problems slowly disappear, but motor retardation with swallowing difficulties and spasticity emerge. Patients usually die at the end of the second or beginning of the third decade of life, although survival into the fourth decade has been reported. Although currently no effective therapy is yet available for MPS III, several promising developments raise hope that therapeutic interventions, halting the devastating mental and behavioural deterioration, might be feasible in the near future.


Preoperative Breast Magnetic Resonance Imaging Use by Breast Density and Family History of Breast Cancer.
Use of preoperative breast magnetic resonance imaging (MRI) among women with a new breast cancer has increased over the past decade. MRI use is more frequent in younger women and those with lobular carcinoma, but associations with breast density and family history of breast cancer are unknown. Data for 3075 women ages >65 years with stage 0-III breast cancer who underwent breast conserving surgery or mastectomy from 2005 to 2010 in the Breast Cancer Surveillance Consortium were linked to administrative claims data to assess associations of preoperative MRI use with mammographic breast density and first-degree family history of breast cancer. Multivariable logistic regression estimated adjusted odds ratios (OR) and 95% confidence intervals (95% CI) for the association of MRI use with breast density and family history, adjusting for woman and tumor characteristics. Overall, preoperative MRI use was 16.4%. The proportion of women receiving breast MRI was similar by breast density (17.6% dense, 16.9% nondense) and family history (17.1% with family history, 16.5% without family history). After adjusting for potential confounders, we found no difference in preoperative MRI use by breast density (OR = 0.95 for dense vs. nondense, 95% CI: 0.73-1.22) or family history (OR = 0.99 for family history vs. none, 95% CI: 0.73-1.32). Among women aged >65 years with breast cancer, having dense breasts or a first-degree relative with breast cancer was not associated with greater preoperative MRI use. This utilization is in keeping with lack of evidence that MRI has higher yield of malignancy in these subgroups.


A new dinuclear Ti(IV)-peroxo-citrate complex from aqueous solutions. Synthetic, structural, and spectroscopic studies in relevance to aqueous titanium(IV)-peroxo-citrate speciation.
The wide use of titanium in applied materials has prompted pertinent studies targeting the requisite chemistry of that metal's biological interactions. In order to understand such interactions as well as the requisite titanium aqueous speciation, we launched investigations on the synthesis and spectroscopic and structural characterization of Ti(IV) species with the physiological citric acid. Aqueous reactions of TiCl(4) with citric acid in the presence of H(2)O(2) and neutralizing ammonia afforded expediently the red crystalline material (NH(4))(4)[Ti(2)(O(2))(2)(C(6)H(4)O(7))(2)].2H(2)O (1). Complex 1 was further characterized by UV-vis, FT-IR, FT- and laser-Raman, NMR, and finally by X-ray crystallography. Compound 1 crystallizes in the monoclinic space group P2(1)/n, with a = 10.360(4) A, b = 10.226(4) A, c = 11.478(6) A, beta = 107.99(2) degrees, V = 1156.6(9) A(3), and Z = 2. The X-ray structure of 1 reveals a dinuclear anionic complex containing a Ti(IV)(2)O(2) core. In that central unit, two fully deprotonated citrate ligands are coordinated to the metal ions through their carboxylate moieties in a monodentate fashion. The central alkoxides serve as bridges to the two titanium ions. Also attached to the Ti(IV)(2)O(2) core are two peroxo ligands each bound in a side-on fashion to the respective metal ions. NH(4)(+) ions neutralize the 4- charge of the anion in 1, further contributing to the stability of the derived lattice through H-bond formation. The structural similarities and differences with congener vanadium(V)-peroxo-citrate complexes may point out potential implications in the chemistry of titanium with physiological ligands, when the former is present in a biologically relevant medium.


A reductionist approach to dissecting grain weight and yield in wheat.
Grain yield is a highly polygenic trait that is influenced by the environment and integrates events throughout the life cycle of a plant. In wheat, the major grain yield components often present compensatory effects among them, which alongside the polyploid nature of wheat, makes their genetic and physiological study challenging. We propose a reductionist and systematic approach as an initial step to understand the gene networks regulating each individual yield component. Here, we focus on grain weight and discuss the importance of examining individual sub-components, not only to help in their genetic dissection, but also to inform our mechanistic understanding of how they interrelate. This knowledge should allow the development of novel combinations, across homoeologs and between complementary modes of action, thereby advancing towards a more integrated strategy for yield improvement. We argue that this will break barriers in terms of phenotypic variation, enhance our understanding of the physiology of yield, and potentially deliver improved on-farm yield.


The effect of oral contraceptive agents on the basal metabolic rate of young women.
The use of oral contraceptive agents by women may be a factor that contributes to the observed inter-individual variability in the BMR. We, therefore, measured the BMR, body build and composition in two groups of young women and also assessed their self-reported level of physical activity. One group had been using oral contraceptive agents for a period of 6 months or more (OCA, n 24), while the other group had never used oral contraceptives (NOCA, n 22). There were no significant differences in age, body build or composition. The absolute BMR in the groups were not significantly different when compared using an unpaired t test (OCA: 5841 (SD 471) v. NOCA: 5633 (SD 615) kJ/d). However, using an analysis of covariance, with either body weight or a combination of fat and fat free mass as covariates, the OCA group had a BMR almost 5% higher than that of the NOCA group (OCA: 5871 v. NOCA: 5601 kJ/d; P = 0.002). When those subjects with high self-reported levels of physical activity were excluded, the difference in BMR between the two groups persisted (P = 0.001). An ANOVA of oral contraceptives use and phase of menstrual cycle showed significant differences in BMR with use of oral contraceptives (P = 0.004) but no difference in BMR between phases of the menstrual cycle. In conclusion, the use of oral contraceptive agents deserves consideration when conducting and analysing data from studies on energy metabolism in young women, as it results in a significantly higher BMR.


A Systematic Review of Carotenoids in the Management of Age-Related Macular Degeneration.
Age-related macular degeneration (AMD) remains a leading cause of modifiable vision loss in older adults. Chronic oxidative injury and compromised antioxidant defenses represent essential drivers in the development of retinal neurodegeneration. Overwhelming free radical species formation results in mitochondrial dysfunction, as well as cellular and metabolic imbalance, which becomes exacerbated with increasing age. Thus, the depletion of systemic antioxidant capacity further proliferates oxidative stress in AMD-affected eyes, resulting in loss of photoreceptors, neuroinflammation, and ultimately atrophy within the retinal tissue. The aim of this systematic review is to examine the neuroprotective potential of the xanthophyll carotenoids lutein, zeaxanthin, and meso-zeaxanthin on retinal neurodegeneration for the purpose of adjunctive nutraceutical strategy in the management of AMD. A comprehensive literature review was performed to retrieve 55 eligible publications, using four database searches from PubMed, Embase, Cochrane Library, and the Web of Science. Epidemiology studies indicated an enhanced risk reduction against late AMD with greater dietary consumption of carotenoids, meanwhile greater concentrations in macular pigment demonstrated significant improvements in visual function among AMD patients. Collectively, evidence strongly suggests that carotenoid vitamin therapies offer remarkable synergic protection in the neurosensory retina, with the potential to serve as adjunctive nutraceutical therapy in the management of established AMD, albeit these benefits may vary among different stages of disease.


The Dorsal Root Ganglion as a Novel Neuromodulatory Target to Evoke Strong and Reproducible Motor Responses in Chronic Motor Complete Spinal Cord Injury: A Case Series of Five Patients.
Current strategies for motor recovery after spinal cord injury (SCI) aim to facilitate motor performance through modulation of afferent input to the spinal cord using epidural electrical stimulation (EES). The dorsal root ganglion (DRG) itself, the first relay station of these afferent inputs, has not yet been targeted for this purpose. The current study aimed to determine whether DRG stimulation can facilitate clinically relevant motor response in motor complete SCI. Five patients with chronic motor complete SCI were implanted with DRG leads placed bilaterally on level L4 during five days. Based on personalized stimulation protocols, we aimed to evoke dynamic (phase 1) and isotonic (phase 2) motor responses in the bilateral quadriceps muscles. On days 1 and 5, EMG-measurements (root mean square [RMS] values) and clinical muscle force measurements (MRC scoring) were used to measure motor responses and their reproducibility. In all patients, DRG-stimulation evoked significant phase 1 and phase 2 motor responses with an MRC ≥4 for all upper leg muscles (rectus femoris, vastus lateralis, vastus medialis, and biceps femoris) (p < 0.05 and p < 0.01, respectively), leading to a knee extension movement strong enough to facilitate assisted weight bearing. No significant differences in RMS values were observed between days 1 and 5 of the study, indicating that motor responses were reproducible. The current paper provides first evidence that bilateral L4 DRG stimulation can evoke reproducible motor responses in the upper leg, sufficient for assisted weight bearing in patients with chronic motor complete SCI. As such, a new target for SCI treatment has surfaced, using existing stimulation devices, making the technique directly clinically accessible.


HIV-1 gp120-specific antibody-dependent cell-mediated cytotoxicity correlates with rate of disease progression.
The Ab-dependent cell-mediated cytotoxicity (ADCC) activity of anti-gp120 Abs in serum from four groups of HIV-1-positive individuals in the Multicenter AIDS Cohort Study was evaluated at several time points over a 10-yr period. HIV-1-positive individuals who progressed to AIDS within 3 yr of seroconversion (rapid progressors) were compared with seroconverters who did not progress to AIDS within 6 yr (nonrapid progressors) and individuals who were seropositive when they entered the study and did not progress to AIDS within 9-10 yr (nonprogressors). At the visit closest to AIDS, rapid progressors had significantly lower titers of Abs that mediate ADCC against HIV-1 gp120 than those of nonrapid progressors at corresponding visits or those of nonprogressors at any visit. Nonprogressors generally had high titers of ADCC Abs at all visits. Differences between ADCC titers of rapid progressors and nonrapid progressors or nonprogressors remained when longitudinal changes within individuals were compared. Among seroconverters who were nonrapid progressors, those with low or declining ADCC titers lost significantly more CD4+ cells during the study than those whose ADCC titers were stable or increasing, even though both groups had similar serum virion RNA levels. This demonstrates that high titers of Abs that mediate ADCC correlate with a successful host defense against AIDS.


Retrograde orotracheal intubation with a double-lumen tube.
Difficult airway management in thoracic surgeries is a peculiar subject due to the demands of monopulmonary ventilation with double-lumen tubes. Flexible bronchoscopy guidance is extremely important, but it is not always available. The objective of this report was to describe a case of retrograde selective orotracheal intubation in the absence of specific endoscopy equipment for the procedure. This is a patient with a history of retosigmoidectomy, admitted for a right thoracotomy for a lung lesion. Preoperative anesthetic evaluation did not reveal any clinical and physical exam particularities. After anesthetic induction and ventilation with face mask, two attempts of orotracheal intubation under direct laryngoscopy were ineffective due to difficult visualization of the vocal folds (Cormack-Lehane grade III). Due to the unavailability of specific material for selective endoscopic intubation it was decided to use the retrograde technique using the double-lumen tube. The patient was extubated in the operating room shortly after the end of the surgery without complications secondary to the alternative technique. Retrograde selective orotracheal intubation is a minimally invasive technique, low cost, safe, and extremely useful whenever flexible bronchoscopy is not available.


Poliovirus vaccine options.
As a result of the success of immunization, indigenous wild poliomyelitis has disappeared from the United States. Of 142 confirmed cases of paralytic poliomyelitis reported in the United States from 1980 to 1996, 134 were classified as vaccine-associated paralytic poliomyelitis (VAPP). Persons with VAPP have a disabling illness, and this has caught the attention of the lay media. The risk of VAPP is one case per 750,000 doses distributed for the first dose of oral poliovirus vaccine (OPV) and one case per 2.4 million doses of OPV distributed overall. Because of this risk, most parents prefer a vaccine schedule that starts with inactivated poliovirus vaccine (IPV), even though extra injections are required. IPV does not cause VAPP. New studies show that high immunization rates can be achieved in disadvantaged populations with a schedule starting with IPV. The American Academy of Family Physicians now recommends that the first two doses of poliovirus vaccine should be IPV; that is, either an all-IPV schedule or a sequential schedule of two doses of IPV followed by two doses of OPV. OPV is no longer recommended for the first two doses and is acceptable only under special circumstances, such as when parents do not accept the recommended number of injections.


Parasite polyclonal activators: new targets for vaccination approaches?
Taking into consideration that the immune response following infection promotes the expansion of lymphocyte clones that are essentially non-specific, ensuring both parasite evasion and persistence inside the host, what would be the major consequences of this polyclonal response to the development of immunopathology? We favor the hypothesis that the polyclonal B cell responses triggered by the infection is responsible of the host susceptibility and is a major contributor to the maintenance of a progressive disease. In particular, the activation of B cells by parasite mitogens would contribute to the class determination of T cell responses and to the inhibition of macrophages - target cells for parasite multiplication and also responsible for parasite clearance. We also envisage that the activation of T cells by parasite 'superantigens', and the ensuing energy and deletion of these cells, processes that are frequently observed, would contribute for the immunosuppression as well as to parasite escape and persistence in the host. We had concentrated our efforts on the study of the non-specific aspects of the immune response following Trypanosoma cruzi infection. We aimed at finding new strategies to modulate and control the mechanisms leading to both the immunosuppression and the development of chronic auto-immunity leading to rational vaccine approaches against parasite infection and immunopathology.


Childhood Sexual Abuse and Fear of Abandonment Moderate the Relation of Intimate Partner Violence to Severity of Dissociation.
Betrayal trauma theory proposes a relation between intimate partner violence (IPV) and dissociation, suggesting that dissociation among victims of IPV may function to restrict awareness of abuse in order to preserve attachments perceived as vital. We investigated two factors that may moderate the relation between IPV and dissociation-childhood sexual abuse (CSA) severity and fear of abandonment-among 348 women currently in a relationship. The relation between frequency of IPV (sexual and physical) and dissociation (amnesia and depersonalization) was moderated by CSA severity and fear of abandonment. Specifically, among women with clinically relevant fear of abandonment, the strength of the relation between IPV and dissociation became stronger as CSA severity increased. This study is the first to demonstrate the moderating roles of fear of abandonment and CSA history in the relation between IPV and dissociation among women. Findings suggest that it may be important to target fear of abandonment in interventions with IPV victims who have a CSA history. Results suggest that fear of abandonment warrants greater attention in research on IPV revictimization.


Cationic ferritin and segmental flow through the trabecular meshwork.
To determine whether segmental labeling by the tracer molecule cationic ferritin (CF) is indicative of preferential patterns of fluid flow in the trabecular meshwork or of differences in cell and extracellular matrix properties. Nonlabeled regions could indicate no fluid entering that area, insufficient perfusion time, or that the cells and extracellular matrix differ in that region and cannot bind CF. Six whole eyes (three normal and three with pseudoexfoliation [PEX]) syndrome were perfused with CF for 30 minutes to 4 hours. Wedges of trabecular meshwork were dissected and some wedges immediately fixed. Adjacent wedges were placed in a CF bath before fixation. Transmission electron microscopy was used to analyze CF labeling. CF increased in the trabecular meshwork with increasing perfusion time. At 30 minutes, CF labeled mainly the uveal and corneoscleral regions. By 4 hours, CF was found diffusely through the meshwork, although a few isolated nonlabeled areas were still present. Wedges immersed in the CF bath showed fewer nonlabeled regions at all time points. Clumps of PEX material labeled more heavily in the periphery than the center, suggesting the clumps were less permeable than surrounding regions. PEX eyes otherwise had similar labeling patterns. Segmental labeling with CF implies regions of preferential flow exist in the meshwork. With increasing perfusion time, there were fewer nonlabeled regions. CF labeling of most regions of bath-immersed tissue suggests that nonlabeled regions do not differ in the characteristics of the cells, but rather that CF does not reach these regions.


Mapping the Solastalgia Literature: A Scoping Review Study.
Solastalgia is a relatively new concept for understanding the links between human and ecosystem health, specifically, the cumulative impacts of climatic and environmental change on mental, emotional, and spiritual health. Given the speed and scale of climate change alongside biodiversity loss, pollution, deforestation, unbridled resource extraction, and other environmental challenges, more and more people will experience solastalgia. This study reviewed 15 years of scholarly literature on solastalgia using a scoping review process. Our goal was to advance conceptual clarity, synthesize the literature, and identify priorities for future research. Four specific questions guided the review process: (1) How is solastalgia conceptualized and applied in the literature?; (2) How is solastalgia experienced and measured in the literature?; (3) How is 'place' understood in the solastalgia literature?; and (4) Does the current body of literature on solastalgia engage with Indigenous worldviews and experiences? Overall, we find there is a need for additional research employing diverse methodologies, across a greater diversity of people and places, and conducted in collaboration with affected populations and potential knowledge, alongside greater attention to the practical implications and applications of solastalgia research. We also call for continued efforts to advance conceptual clarity and theoretical foundations. Key outcomes of this study include our use of the landscape construct in relation to solastalgia and a call to better understand Indigenous peoples' lived experiences of landscape transformation and degradation in the context of historical traumas.


An objective approach to dry eye disease severity.
A prospective, multisite clinical study (10 sites in the European Union and the United States) evaluated the clinical utility of commonly used tests and tear osmolarity for assessing dry eye disease severity. Three hundred fourteen consecutive subjects between the ages of 18 and 82 years were recruited from the general patient population, 299 of which qualified with complete datasets. Osmolarity testing, Schirmer test without anesthesia, tear film breakup time (TBUT), corneal staining, meibomian dysfunction assessment, and conjunctival staining were performed bilaterally. A symptom questionnaire, the Ocular Surface Disease Index (OSDI), was also administered to each patient. Distributions of clinical signs and symptoms against a continuous composite severity index were evaluated. Osmolarity was found to have the highest correlation coefficient to disease severity (r(2) = 0.55), followed by conjunctival staining (r(2) = 0.47), corneal staining (r(2) = 0.43), OSDI (r(2) = 0.41), meibomian score (r(2) = 0.37), TBUT (r(2) = 0.30), and Schirmer result (r(2) = 0.17). A comparison of standard threshold-based classification with the composite severity index revealed significant overlap between the disease severities of prospectively defined normal and dry eye groups. Fully 63% of the subjects were found to be poorly classified by combinations of clinical thresholds. Tear film osmolarity was found to be the single best marker of disease severity across normal, mild/moderate, and severe categories. Other tests were found to be informative in the more severe forms of disease; thus, clinical judgment remains an important element in the clinical assessment of dry eye severity. The results also indicate that the initiation and progression of dry eye is multifactorial and supports the rationale for redefining severity on the basis of a continuum of clinical signs. (ClinicalTrials.gov number, NCT00848198.).


Costimulation blockade: current perspectives and implications for therapy.
T cells must be activated before they can elicit damage to allografts, through interaction of their T cell receptor (TCR) with peptide-MHC complex and through accessory molecules. Signaling through accessory molecules or costimulatory molecules is a critical way for the immune system to fine tune T cell activation. An emerging therapeutic strategy is to target selective molecules involved in the process of T cell activation using biologic agents, which do not impact TCR signaling, thus only manipulating the T cells, which recognize alloantigen. Costimulatory receptors and their ligands are attractive targets for this strategy and could be used both to prevent acute graft rejection as well as for maintenance immunosuppression. Therapeutic agents targeting costimulatory molecules, notably belatacept, have made the progression from the bench, through nonhuman primate studies and into the clinic. This overview describes some of the most common costimulatory molecules, their role in T cell activation, and the development of reagents, which target these pathways and their efficacy in transplantation.


Relationship between pigment production and haemolysin formation by Lancefield group B streptococci.
Group B streptococci produce both a pigment and a haemolysin. The requirements of group B streptococci for the formation and release of pigment and haemolysin are similar and have been examined to extend observations on the relationship between the two products. The amount of pigment and haemolysin extractable from actively metabolising washed-cell suspensions of group B streptococci varied with the atmosphere of incubation, the pH at which the extraction was carried out and the presence of Mg2+ ions. Both pigment and haemolysin were produced in significant amounts in all phases of the growth cycle. When conditions were established for obtaining maximum yields of haemolysin, its production correlated closely with pigment yields, but pigment did not function as a carrier for haemolysin. Formation of pigment, but not of haemolysin, increased in the presence of trimethoprim or higher concentrations of glucose. The composition of pigment produced in different conditions differed qualitatively and different strains of group B streptococci formed pigment of different appearance, suggesting that group B streptococcal pigment is composed of several different substances.


Knowledge, attitude and practice of pediatric critical care nurses towards pain: survey in a developing country setting.
Nurses' knowledge, sensitivity and attitudes about pain in children and its management affect their response and therefore management of pediatric pain. Children in critical care units undergo more painful procedures than those in general wards. To study the knowledge, attitude and practice of nursing personnel catering to critically ill children in a developing country. Prospective questionnaire-based survey. The survey was carried out in a tertiary care teaching hospital on nursing personnel in three pediatric/neonatal intensive care units. The domains studied were: i. Training and experience, ii. Knowledge of pediatric pain, iii. Individual attitude towards pain in children, iv. Personal practice(s) for pain alleviation, v. Pain assessment, and vi. Non-pharmacological measures adopted. Descriptive statistics and logistic regression. Of the 81 nursing personnel working in the three critical care units, 56 (69.1%) responded to the questionnaire. Only one-third of them had received formal training in pediatric nursing. Fifty percent of the respondents felt that infants perceive less pain than adults. Training in pediatric nursing was a significant contributing factor in the domain of knowledge (P=0.03). Restraint and distraction were the common modalities employed to facilitate painful procedures. Scientific approaches like eutectic mixture of local anesthetic and the judicious use of sedatives were not adopted routinely. Observing a child's face and posture were widely used parameters to assess pain (83%). None of the three critical care areas used a scoring system to assess pain. There are several lacunae in the knowledge and practice of nurses in developing countries which need to be improved by training.


Synergy between inhibitors of androgen receptor and MEK has therapeutic implications in estrogen receptor-negative breast cancer.
Estrogen receptor-negative (ER-) breast cancer is a heterogeneous disease with limited therapeutic options. The molecular apocrine subtype constitutes 50% of ER-tumors and is characterized by overexpression of steroid response genes including androgen receptor (AR). We have recently identified a positive feedback loop between the AR and extracellular signal-regulated kinase (ERK) signaling pathways in the molecular apocrine subtype. In this feedback loop, AR regulates ERK phosphorylation through the mediation of ErbB2 and, in turn, ERK-CREB1 signaling regulates the transcription of AR in molecular apocrine cells. In this study, we investigated the therapeutic implications of the AR-ERK feedback loop in molecular apocrine breast cancer. We examined a synergy between the AR inhibitor flutamide and the MEK inhibitor CI-1040 in the molecular apocrine cell lines MDA-MB-453, HCC-1954 and HCC-202 using MTT cell viability and annexin V apoptosis assays. Synergy was measured using the combination index (CI) method. Furthermore, we examined in vivo synergy between flutamide and the MEK inhibitor PD0325901 in a xenograft model of the molecular apocrine subtype. The effects of in vivo therapies on tumor growth, cell proliferation and angiogenesis were assessed. We demonstrate synergistic CI values for combination therapy with flutamide and CI-1040 across three molecular apocrine cell lines at four dose combinations using both cell viability and apoptosis assays. Furthermore, we show in vivo that combination therapy with flutamide and MEK inhibitor PD0325901 has a significantly higher therapeutic efficacy in reducing tumor growth, cellular proliferation and angiogenesis than monotherapy with these agents. Moreover, our data suggested that flutamide and CI-1040 have synergy in trastuzumab resistance models of the molecular apocrine subtype. Notably, the therapeutic effect of combination therapy in trastuzumab-resistant cells was associated with the abrogation of an increased level of ERK phosphorylation that was developed in the process of trastuzumab resistance. In this study, we demonstrate in vitro and in vivo synergies between AR and MEK inhibitors in molecular apocrine breast cancer. Furthermore, we show that combination therapy with these inhibitors can overcome trastuzumab resistance in molecular apocrine cells. Therefore, a combination therapy strategy with AR and MEK inhibitors may provide an attractive therapeutic option for the ER-/AR+ subtype of breast cancer.


A bioelectrochemical approach to characterize extracellular electron transfer by Synechocystis sp. PCC6803.
Biophotovoltaic devices employ photosynthetic organisms at the anode of a microbial fuel cell to generate electrical power. Although a range of cyanobacteria and algae have been shown to generate photocurrent in devices of a multitude of architectures, mechanistic understanding of extracellular electron transfer by phototrophs remains minimal. Here we describe a mediatorless bioelectrochemical device to measure the electrogenic output of a planktonically grown cyanobacterium, Synechocystis sp. PCC6803. Light dependent production of current is measured, and its magnitude is shown to scale with microbial cell concentration and light intensity. Bioelectrochemical characterization of a Synechocystis mutant lacking Photosystem II demonstrates conclusively that production of the majority of photocurrent requires a functional water splitting aparatus and electrons are likely ultimately derived from water. This shows the potential of the device to rapidly and quantitatively characterize photocurrent production by genetically modified strains, an approach that can be used in future studies to delineate the mechanisms of cyanobacterial extracellular electron transport.


The systemic response to CNS injury.
Inflammation within the brain or spinal cord has the capacity to damage neurons and is known to contribute to long-term disability in a spectrum of central nervous system (CNS) pathologies. However, there is a more profound increase in the recruitment of potentially damaging populations of leukocytes to the spinal cord than to the brain after equivalent injuries. Increased levels of inflammatory cytokines and chemokines in the spinal cord underpin this dissimilarity after injury, which also appears to be very sensitive to processes that operate within organs distant from the primary injury site such as the liver, lung and spleen. Indeed, CNS injury per se can generate profound changes in gene expression and the cellularity of these organs, which, as a consequence, gives rise to secondary organ damage. Our understanding of the local inflammatory processes that can damage neurons is becoming clearer, but our understanding of how the peripheral immune system coordinates the response to CNS injury and how any concomitant infections or injury might impact on the outcome of CNS injury is not so well developed. It is clear that the orientation of the response to peripheral challenges, be it a pro- or anti-inflammatory effect, appears to be dependent on the nature and timing of events. Here, the importance of the inter-relationship between inflammation in the CNS and the consequent inflammatory response in peripheral tissues is highlighted.


Effects of repeated prenatal ultrasound examinations on childhood outcome up to 8 years of age: follow-up of a randomised controlled trial.
Despite the widespread use of prenatal ultrasound studies, there are no published data from randomised controlled trials describing childhood outcomes that might be influenced by repeated ultrasound exposures. We previously undertook a randomised controlled trial to assess the effects of multiple studies on pregnancy and childhood outcomes and reported that those pregnancies allocated to receive multiple examinations had an unexplained and significant increase in the proportion of growth restricted newborns. Our aim was to investigate the possible effects of multiple prenatal ultrasound scans on growth and development in childhood. Here, we provide follow-up data of the childrens' development. Physical and developmental assessments were done on children whose pregnant mothers had been allocated at random to a protocol of five studies of ultrasound imaging and umbilical artery Doppler flow velocity waveform between 18 and 38 weeks' gestation (intensive group n=1490) or a single imaging study at 18 weeks' gestation (regular group n=1477). We used generalised logistic and linear regression models to assess the group differences in developmental and growth outcomes over time. Primary data analysis was done by intention-to-treat. Examinations were done at 1, 2, 3, 5, and 8 years of age on children born without congenital abnormalities and from singleton pregnancies (intensive group n=1362, regular group n=1352). The follow-up rate at 1 year was 85% (2310/2714) and at 8 years was 75% (2042/2714). By 1 year of age and thereafter, physical sizes were similar in the two groups. There were no significant differences indicating deleterious effects of multiple ultrasound studies at any age as measured by standard tests of childhood speech, language, behaviour, and neurological development. Exposure to multiple prenatal ultrasound examinations from 18 weeks' gestation onwards might be associated with a small effect on fetal growth but is followed in childhood by growth and measures of developmental outcome similar to those in children who had received a single prenatal scan.


ATM activation in hypoxia - causes and consequences.
The DNA damage response is a complex signaling cascade that is triggered by cellular stress. This response is essential for the maintenance of genomic integrity and is considered to act as a barrier to the early stages of tumorigenesis. The integral role of ataxia telangiectasia mutated (ATM) kinase in the response to DNA damaging agents is well characterized; however, ATM can also be activated by non-DNA damaging agents. In fact, much has been learnt recently about the mechanism of ATM activation in response to physiologic stresses such as hypoxia that do not induce DNA damage. Regions of low oxygen concentrations that occur in solid tumors are associated with a poor prognostic outcome irrespective of treatment modality. Severe levels of hypoxia induce replication stress and trigger the activation of DNA damage response pathways including ataxia telangiectasia and Rad3-related (ATR)- and ATM-mediated signaling. In this review, we discuss hypoxia-driven ATM signaling and the possible contribution of ATM activation in this context to tumorigenesis.


Fabrication of high quality, ultra-long fiber Bragg gratings: up to 2 million periods in phase.
The fabrication and characterization of high quality ultra-long (up to 1m) fiber Bragg gratings (FBGs) is reported. A moving phase mask and an electro-optic phase-modulation (EOPM) based interferometer are used with a high precision 1-meter long translation stage and compared. A novel interferometer position feedback scheme to simplify the fabrication process is proposed and analyzed. The ultra-long uniform FBGs show near perfect characteristics of a few picometers bandwidth, symmetrical, near theory-matching group-delay and transmission spectra. Grating characterization using optical backscattering reflectometry and chirped FBGs are also demonstrated. Limitations of the schemes are discussed.


Reciprocal activation of macrophages and breast carcinoma cells by nitric oxide and colony-stimulating factor-1.
Induction of inducible nitric oxide synthase (iNOS) gene expression, nitric oxide (NO) production and migration of RAW264.7 macrophages by coculture with breast cancer MDA-MB-231 cells or the addition of conditioned medium derived from MDA-MB-231 cells (MDA-CM) was identified. Increased iNOS/NO induction and migration of macrophages by MDA-CM were significantly blocked by adding the c-Jun-N-terminal protein kinase (JNK) inhibitor, SP600125, the nuclear factor-kappa B (NF-κB) inhibitor, BAY117082 and pyrrolidine dithiocarbamic acid and a dominant-negative JNK. The addition of an NO donor, Diethylenetriamine-NONOate, significantly activated expressions of MMP-9 and VEGF-A genes in breast carcinoma MDA-MD-231 cells and invasion of MDA-MB-231 cells in coculture with RAW264.7 macrophages as determined using Transwell systems, but that was inhibited by adding SP600125, BAY117082 and the nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester. Induction of heme oxygenase-1 in macrophages reduced MDA-CM-induced iNOS/NO, JNK and NF-κB activations in accordance with inhibiting VEGF-A and MMP-9 gene expressions by MDA-MB-231 cells via Transwell assays. Furthermore, VEGF, sRANKL, TNF-α, IL-1α, TGF-β, CSF-1 and MCP-1 were applied, and CSF-1 showed the most potent stimulation of iNOS/NO production and migration of macrophages. MCF-7 cells with lower CSF-1 expression than MDA-MB-231 cells showed a poor stimulatory effect on iNOS/NO production and migration of macrophages. Neutralization of CSF-1 in MDA-CM using CSF-1 antibody inhibited MDA-CM-induced iNOS protein expression and migration of macrophages, and CSF-1-induced iNOS protein and migration was blocked by adding JNK inhibitor SP and NF-κB inhibitor BAY. The reciprocal activation of breast cancer and macrophages via NO-CSF-1 is first elucidated herein.


Expression of immunoreactive matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases in human normal livers and primary liver tumors.
Matrix metalloproteinases (MMPs) play an important role in cancer cell invasion by degrading extracellular matrix proteins. However, little is known about the in situ expression of MMP in human normal livers and primary liver tumors. In this study, we therefore examined the in situ expression of immunoreactive MMP and tissue inhibitors of MMP (TIMP) in 10 normal livers, 11 surgically resected intrahepatic cholangiocarcinomas (CCs), and 6 surgically resected hepatocellular carcinomas (HCCs). In normal livers, MMP and TIMP were infrequently and faintly expressed in bile ducts, but were not expressed in hepatocytes. In the 11 CCs, MMP-1, MMP-2, MMP3, MMP-9, TIMP-1, and TIMP-2 were expressed in tumor cells and/or tumor stroma in 11 (100%), 5 (45%), 8 (73%), 3 (27%), 9 (82%), and 9 (82%), respectively. The expression of MMP and TIMP in tumor cells was located in the cytoplasm with a diffuse or granular pattern; that in the tumor stroma was situated in fibroblasts, leukocytes, and extracellular matrix. Their expression was stronger in CC cases with severe invasion than in CC cases with mild invasion. In contrast, MMP and TIMP were not expressed in any cases of HCC. These results show that intrahepatic bile duct cells may neoexpress or overexpress MMP and TIMP after malignant transformation but that hepatocytes do not, and suggest that MMP and TIMP play an important role in CC cell invasion by degrading extracellular matrix proteins.


Matrine inhibits proliferation and induces apoptosis of pancreatic cancer cells in vitro and in vivo.
Matrine, an alkaloid extracted from a Chinese herb, Sophora flavescens AIT., has exhibited anti-proliferative and pro-apoptotic abilities against various types of cancer cells. This study aims to investigate its anti-cancer activity and underlying mechanisms in human pancreatic cancer cells in vitro and in vivo. Human BxPC-3 and PANC-1 pancreatic cancer cells, and human HL-7702 liver cells were incubated with matrine at different concentrations. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and cell apoptosis, by flow cytometry. Subcutaneous BxPC-3 xenograft tumors were established in nude BALB/c mice, and matrine was intraperitoneally (i.p.) administered. The tumors were monitored and harvested. Tumor sections were immunostained with an anti-Ki-67 antibody (Ab) to examine cell proliferation, or stained with terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) to evaluate in situ cell apoptosis. The expression of proliferating cell nuclear antigen (PCNA) and several apoptosis-related proteins in cells and tumor tissues were evaluated by Western blot analysis. In in vitro assays, matrine inhibited cell viability by downregulating the expression of PCNA, and induced cell apoptosis by reducing the ratio of Bcl-2/Bax, upregulating Fas, and increasing activation of caspases-8,-3 and -9, in a dose-dependent manner. Administration of matrine inhibited tumor growth in a dose-dependent manner, and regulated tumoral gene expression consistent with the in vitro results. But matrine had no significant effects on the viability of HL-7702 cells or the bodyweight of mice compared to controls. These results indicate matrine may be a potential and promising agent of natural resource to treat pancreatic cancer.


Making reviewers visible: openness, accountability, and credit.
Anonymity for peer reviewers remains the overwhelming norm within biomedical journals. While acknowledging that open review is not without challenges, this article presents 4 key arguments in its favor: (1) ethical superiority, (2) lack of important adverse effects, (3) feasibility in practice, and (4) potential to balance greater accountability for reviewers with credit for the work they do. Barriers to more widespread use of open review include conservatism within the research community and the fact that openness makes editors publicly responsible for their choice of reviewers and their interpretation of reviewers' comments. Forces for change include the growing use of preprint servers combined with open commentary. I look forward to a time when open commentary and review replace the current, flawed system of closed prepublication peer review and its false reassurances about the reliability of what is published.


Effects of glucose-insulin-potassium on myocardial substrate availability and utilization in stable coronary artery disease. Studies on myocardial carbohydrate, lipid and oxygen arterial-coronary sinus differences in patients with coronary artery disease.
To assess the metabolic effects of myocardial substrate alteration in patients with coronary artery disease, glucose-insulin-potassium solution was administered intravenously for 30 minutes to 14 men with stable angiographically documented coronary artery disease. The glucose-insulin-potassium solution (300 g of glucose, 50 units of regular insulin and 80 mEq of potassium chloride per liter of water) was infused at a constant rate in each patient, but individual infusion rates ranged from 0.013 to 0.032 ml/kg per min (4 to 10 mg glucose/kg per min) in the 14 patients. Simultaneous arterial and coronary sinus samples were obtained at 15 minute intervals during a stable 30 minute control period and again at 15 minute intervals during the infusion; samples were assayed for glucose, lactate, free fatty acid and oxygen content. In all 14 patients, during the glucose-insulin-potassium infusion, arterial glucose and lactate increased and arterial free fatty acid levels fell; the magnitude of the changes in arterial lactate and free fatty acids was related to the rate of infusion. Arterial-coronary sinus differences (A-Cs) for glucose, lactate and free fatty acid levels correlated with the arterial concentrations of these substrates (r = 0.66, 0.87 and 0.79, respectively). Regression analyses demonstrated myocardial thresholds for the uptake of these substrates as follows: glucose 79 mg/100 ml; lactate 300 mu mole/liter; and free fatty acids 100 to 200 mu Eq/liter. Finally and most importantly, the reduction in A-Cs oxygen values after glucose-insulin-potassium infusion correlated with the reduction in A-Cs free fatty acid levels (r = 0.64, P less than 0.0001). This observation suggests that, in patients with coronary artery disease, glucose-insulin-potassium infusion may significantly diminish myocardial oxygen requirements by reduction of myocardial free fatty acid utilization and simultaneous enhancement of myocardial carbohydrate utilization. Myocardial substrate availability may be an important determinant of myocardial oxygen demand in patients with coronary artery disease. Infusion of glucose-insulin-potassium solution has the potential to alter myocardial substrate availability, thus improving the balance between myocardial oxygen demand and supply.


Can becoming a leader change your personality? An investigation with two longitudinal studies from a role-based perspective.
Organizational research has predominantly adopted the classic dispositional perspective to understand the importance of personality traits in shaping work outcomes. However, the burgeoning literature in personality psychology has documented that personality traits, although relatively stable, are able to develop throughout one's whole adulthood. A crucial force driving adult personality development is transition into novel work roles. In this article, we introduce a dynamic, role-based perspective on the adaptive nature of personality during the transition from the role of employee to that of leader (i.e., leadership emergence). We argue that during such role transitions, individuals will experience increases in job role demands, a crucial manifestation of role expectations, which in turn may foster growth in conscientiousness and emotional stability. We tested these hypotheses in two 3-wave longitudinal studies using a quasi-experimental design. We compared the personality development of 2 groups of individuals (1 group promoted from employees into leadership roles and the other remaining as employees over time), matched via the propensity score matching approach. The convergent results of latent growth curve modeling from the 2 studies support our hypotheses regarding the relationship between becoming a leader and subsequent small, but substantial increases in conscientiousness over time and the mediating role of job role demands. The relationship between becoming a leader and change of emotional stability was not significant. This research showcases the prominence of examining and cultivating personality development for organizational research and practice. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


The effect of cocoa procyanidins on the transcription and secretion of interleukin 1 beta in peripheral blood mononuclear cells.
Recent data has demonstrated that cacao liquor polyphenols (procyanidins) have antioxidant activity, inhibit mRNA expression of interleukin-2 and are potent inhibitors of acute inflammation. Given the widespread ingestion of cocoa in many cultures, we investigated whether cocoa, in its isolated procyanidin fractions (monomer through decamer), would modulate synthesis of the pro-inflammatory cytokine, interleukin-1 beta. Both resting and phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) were investigated at the levels of transcription and protein secretion. Individual cocoa fractions were shown to augment constitutive IL-1 beta gene expression, although values varied between subjects. Interestingly, the smaller fractions of cocoa (monomer-tetramer) consistently reduced IL-1 beta expression of PHA-stimulated cells by 1-15%, while the larger oligomers (pentamer-decamer) increased expression by 4-52%. These data, observed at the transcription level, were reflected in protein levels in PHA-induced PBMC. The presence or absence of PHA did not alter the effects of the cocoa procyanidins with the exception of the pentamer. This study offers additional data for the consideration of the health-benefits of dietary polyphenols from a wide variety of foods, including those benefits associated specifically with cocoa and chocolate consumption.


3dRS, a Web-Based Tool to Share Interactive Representations of 3D Biomolecular Structures and Molecular Dynamics Trajectories.
3D Representation Sharing (3dRS) is a web-based tool designed to share biomolecular structure representations, including 4D ensembles derived from Molecular Dynamics (MD) trajectories. The server offers a team working in different locations a single URL to share and discuss structural data in an interactive fashion, with the possibility to use it as a live figure for scientific papers. The web tool allows an easy upload of structures and trajectories in different formats. The 3D representation, powered by NGL viewer, offers an interactive display with smooth visualization in modern web browsers. Multiple structures can be loaded and superposed in the same scene. 1D sequences from the loaded structures are presented and linked to the 3D representation. Multiple, pre-defined 3D molecular representations are available. The powerful NGL selection syntax allows the definition of molecular regions that can be then displayed using different representations. Important descriptors such as distances or interactions can be easily added into the representation. Trajectory frames can be explored using a common video player control panel. Trajectories are efficiently stored and transferred to the NGL viewer thanks to an MDsrv-based data streaming. The server design offers all functionalities in one single web page, with a curated user experience, involving a minimum learning curve. Extended documentation is available, including a gallery with a collection of scenes. The server requires no registration and is available at https://mmb.irbbarcelona.org/3dRS.


Improvement of chemotherapy efficacy by inactivation of a DNA-repair pathway.
Tumour resistance and dose-limiting toxic effects restrict treatment with most chemotherapeutic drugs. Elucidation of the mechanisms of these effects could permit the development of ways to improve the effectiveness of currently used agents until better therapeutic agents are developed. Several types of alkylating agents are used in the treatment of cancer. The DNA repair protein, O6-alkylguanine-DNA alkyltransferase (ATase) is an important cellular resistance mechanism to one class of alkylating agents. This enzyme removes potentially lethal damage from DNA and experiments in vitro and in vivo have shown that its inactivation can reverse resistance to such agents. Clinical trials of drugs that inactivate ATase are underway and early results indicate that they are active in tumour tissues. However, the ATase present in normal tissues, particularly bone marrow, is also inactivated, necessitating a reduction in the dose of alkylating agent. An important question is whether, in the absence of any tumour-specific delivery strategy, such drugs will improve therapeutic effectiveness; initial reports are not promising.


p53-mediated hematopoietic stem and progenitor cell competition.
Cell competition was originally described in Drosophila as a process for selection of the fittest cells. It is likely to play an important role in tissue homeostasis in all metazoans, but little is known about its role and regulation in mammals. By using genetic mosaic mouse models and bone marrow chimeras, we describe here a form of cell competition that selects for the least damaged cells. This competition is controlled by p53 but is distinct from the classical p53-mediated DNA damage response: it persists for months, is specific to the hematopoietic stem and progenitor cells, and depends on the relative rather than absolute level of p53 in competing cells. The competition appears to be mediated by a non-cell-autonomous induction of growth arrest and senescence-related gene expression in outcompeted cells with higher p53 activity. p53-mediated cell competition of this type could potentially contribute to the clonal expansion of incipient cancer cells.


Risk factors for stress fractures in track and field athletes. A twelve-month prospective study.
The aim of this 12-month prospective study was to investigate risk factors for stress fractures in a cohort of 53 female and 58 male track and field athletes, aged 17 to 26 years. Total bone mineral content, regional bone density, and soft tissue composition were measured using dual-energy x-ray absorptiometry and anthropometric techniques. Menstrual characteristics, current dietary intake, and training were assessed using questionnaires. A clinical biomechanical assessment was performed by a physical therapist. The incidence of stress fractures during the study was 21.1% with most injuries located in the tibia. Of the risk factors evaluated, none was able to predict the occurrence of stress fractures in men. However, in female athletes, significant risk factors included lower bone density, a history of menstrual disturbance, less lean mass in the lower limb, a discrepancy in leg length, and a lower fat diet. Multiple logistic regression revealed that age of menarche and calf girth were the best independent predictors of stress fractures in women. This bivariate model correctly assigned 80% of the female athletes into their respective stress fracture or nonstress fracture groups. These results suggest that it may be possible to identify female athletes most at risk for this overuse bone injury.


Asymmetry analysis in rodent cerebral ischemia models.
An automated method for identification and segmentation of acute/subacute ischemic stroke, using the inherent bi-fold symmetry in brain images, is presented. An accurate and automated method for localization of acute ischemic stroke could provide physicians with a mechanism for early detection and potentially faster delivery of effective stroke therapy. Segmentation of ischemic stroke was performed on magnetic resonance (MR) images of subacute rodent cerebral ischemia. Eight adult male Wistar rats weighing 225-300 g were anesthetized with halothane in a mix of 70% nitrous oxide/30% oxygen. Animal core temperature was maintained at 37 degrees C during the entire surgical procedure, including occlusion of the middle cerebral artery (MCA) and the 90-minute post-reperfusion period. To confirm cerebral ischemia, transcranial measurements of cerebral blood flow were performed with laser-Doppler flowmetry, using 15-mm flexible fiberoptic Doppler probes attached to the skull over the MCA territory. Animal MR scans were performed at 1.5 T using a knee coil. Three experts performed manual tracing of the stroke regions for each rat, using the histologic-stained slices to guide delineation of stroke regions. A strict tracing protocol was followed that included multiple (three) tracings of each stroke region. The volumetric MR image data were processed for each rat by computing the axis of symmetry and extracting statistical dissimilarities. A nonparametric Wilcoxon rank sum test operating on paired windows in opposing hemispheres identified seeds in the pixels exhibiting statistically significant bi-fold mirror asymmetry. Two brain reference maps were used for analysis: an absolute difference map (ADM) and a statistical difference map (SDM). Although an ADM simply displays the absolute difference by subtracting one brain hemisphere from its reflection, SDM highlights regions by labeling pixels exhibiting statistically significant asymmetry. To assess the accuracy of the proposed segmentation method, the surrogate ground truth (the stroke tracing data) was compared to the results of our proposed automated segmentation algorithm. Three accuracy segmentation metrics were utilized: true-positive volume fraction (TPVF), false-positive volume fraction (FPVF), and false-negative volume fraction (FNVF). The mean value of the TPVF for our segmentation method was 0.8877; 95% CI 0.7254 to 1.0500; the mean FPVF was 0.3370, 95% CI -0.0893 to 0.7633; the mean FNVF was 0.1122, 95% CI -0.0502 to 0.2747. Unlike most segmentation methods that require some degree of manual intervention, our segmentation algorithm is fully automated and highly accurate in identifying regions of brain asymmetry. This approach is attractive for numerous neurologic applications where the operator's intervention should be minimal or null.


Prevalence of noise-induced hearing loss in student musicians.
This study describes the prevalence and characteristics of noise-induced hearing loss (NIHL) in student musicians (N = 329) aged 18-25 years. Students completed a questionnaire regarding exposures before a hearing assessment. NIHL was defined by the presence of a notch 15 dB in depth at 4000 or 6000 Hz relative to the best preceding threshold. Overall prevalence of NIHL was 45%, with 78% of notches occurring at 6000 Hz. The proportion of the total population with bilateral notching at any frequency was 11.5%, mostly occurring at 6000 Hz. There was a significant increase in the frequency of notching in students who reported more than two hours per day of personal practice. There were no significant associations for instrument group or other noise exposures. The data suggest that susceptibility to NIHL among students of music is not uniform and cannot be ascribed solely to the instrument played and other exposures. Students with bilateral losses tend to have deeper notches and may represent a group that has an inherent predisposition to NIHL.


Curled cation structures accelerate the dynamics of ionic liquids.
Ionic liquids are modern liquid materials with potential and actual implementation in many advanced technologies. They combine many favourable and modifiable properties but have a major inherent drawback compared to molecular liquids - slower dynamics. In previous studies we found that the dynamics of ionic liquids are significantly accelerated by the introduction of multiple ether side chains into the cations. However, the origin of the improved transport properties, whether as a result of the altered cation conformation or due to the absence of nanostructuring within the liquid as a result of the higher polarity of the ether chains, remained to be clarified. Therefore, we prepared two novel sets of methylammonium based ionic liquids; one set with three ether substituents and another set with three butyl side chains, in order to compare their dynamic properties and liquid structures. Using a range of anions, we show that the dynamics of the ether-substituted cations are systematically and distinctly accelerated. Liquefaction temperatures are lowered and fragilities increased, while at the same time cation-anion distances are slightly larger for the alkylated samples. Furthermore, pronounced liquid nanostructures were not observed. Molecular dynamics simulations demonstrate that the origin of the altered properties of the ether substituted ionic liquids is primarily due to a curled ether chain conformation, in contrast to the alkylated cations where the alkyl chains retain a linear conformation. Thus, the observed structure-property relations can be explained by changes in the geometric shape of the cations, rather than by the absence of a liquid nanostructure. Application of quantum chemical calculations to a simplified model system revealed that intramolecular hydrogen-bonding is responsible for approximately half of the stabilisation of the curled ether-cations, whereas the other half stems from non-specific long-range interactions. These findings give more detailed insights into the structure-property relations of ionic liquids and will guide the development of ionic liquids that do not suffer from slow dynamics.