Theoretical modulation of singlet/triplet chemiexcitation of chemiluminescent imidazopyrazinone dioxetanone via C8-substitution.
Coelenterazine, a member of the imidazopyrazinone class of chemiluminescent substrates, presents significant potential as a dynamic probe of reactive oxygen species in a biological environment, such as a superoxide anion, in which these species are important in cellular biology and pathology. The objective of the current study was to understand in what way the efficiency of singlet and triplet chemiexcitation could be modulated, towards a more efficient use of imidazopyrazinone-based compounds as dynamic chemiluminescent probes. To this end the thermolysis of imidazopyrazinone dioxetanone, substituted at the C8-position with electron-donating or electron-withdrawing groups, was characterized with a theoretical approach based on density functional theory. Substituents with different electron-donating/withdrawing characters have only a limited effect on the singlet chemiexcitation of anionic dioxetanone. For neutral dioxetanone, both electron-withdrawing and weak electron-donating substituents increase singlet chemiexcitation, to the contrary of strong electron-donating groups. During their thermolysis reaction, all molecules presented regions of degeneracy with triplet states, thereby indicating the possibility of triplet chemiexcitation.


Comparison of condylar position in orthognathic surgery cases treated with virtual surgical planning vs. conventional model planning.
To assess the effect of virtual surgical planning (VSP) on the accuracy of condylar seating in surgical cases involving Bilateral Sagittal Split Osteotomy (BSSO). An additional aim was to quantify differences between actual surgical outcome and planned virtual outcome. A private practice in Charlotte North Carolina. Twenty-five consecutively operated subjects prepared with VSP and twenty consecutively subjects operated with conventional surgery planning were enrolled. Cone beam computed tomography (CBCT) scans were obtained before surgery (T1) and post-surgery after splint removal (T2). The T1 and T2 CBCTs were superimposed on the anterior cranial base using voxel-based registration. Three dimensional (3D) models were built and oriented to a 3D Frankfurt horizontal. Displacement of corresponding condylar landmarks was measured. No significant difference was found between the VSP and conventional group in the magnitude of translation or rotation of the condyles following surgery. VSP surgical outcomes differed significantly from predicted positions at all landmarks. Virtual surgical planning does not prevent changes to condylar position as a result of surgery. Actual surgical results differed significantly from VSP-predicted outcomes. Virtual surgical planning did not reduce the changes to condylar position and angulation that resulted from conventionally planned orthognathic surgery.


Trade-offs between location and waiting times in the provision of health care: the case of elective surgery on the Isle of Wight.
Recent UK government documents have stated that, within the National Health Service, consumers should be more involved in decision-making. This study considered the technique of discrete choice conjoint analysis (DCCA) for eliciting community views regarding the importance of reducing waiting times. More specifically, the study aimed to establish whether residents living on the Isle of Wight are prepared to travel to the mainland for elective surgery where waiting times are shorter but travel costs may be greater, and, further, if residents are willing to travel, what reduction in waiting time and increase in travel costs would be acceptable. A DCCA questionnaire was sent to a random sample of 1,000 individuals living on the Isle of Wight. Seventy-eight per cent of respondents were willing to travel to the mainland for elective surgery. Of these, 48 per cent always chose the mainland and 30 per cent traded between island and mainland, depending on the levels of waiting time and travel costs. Whereas 'traders' preferred the Isle of Wight, they would forgo their preferred location if waiting times were reduced by at least 3.9 months, and they were willing to pay over 12 for a 1 month reduction in waiting time. Different combinations of waiting time reductions and travel cost increases that were acceptable to traders were estimated. Carrying out elective surgery on the mainland offers the potential to reduce waiting lists for island residents. The majority of respondents would be willing to travel to the mainland. However, one-fifth of respondents were not willing to travel to the mainland for elective surgery. The preferences of this group are important in policy decisions. More generally, the paper demonstrates the potential application of DCCA to public health issues.


<Editors' Choice> Ultrasound measurement of thigh muscle thickness for assessment of sarcopenia.
Our aim of this study is to compare the thigh muscle thickness measurements obtained using ultrasound and bioelectrical impedance analysis (BIA) methods, and to investigate the validity and cutoff value of the ultrasonography. We analyzed a total of 201 participants (99 male and 102 female participants, mean age, 66.2 years) participated in the annual health checkup in the Yakumo Study, 2014. Thigh muscle thickness (TMT, sum of the rectus femoris and vastus intermedius muscle thickness) was measured using ultrasound at mid-thigh in the sitting position. Appendicular skeletal muscle mass (aSMI) was measured using BIA. Cutoff value of TMT was determined through the receiver operating characteristic analysis. We defined sarcopenia with the diagnostic algorithm of Asian Working Group for Sarcopenia. TMT was significantly reduced in subject with sarcopenia than in those without sarcopenia in both gender. Muscle measurements obtained using the BIA methods (aSMI) and ultrasound methods (TMT) showed a significant correlation, with a correlation coefficient of 0.38 (P < 0.001). Cutoff value, sensitivity, and specificity of TMT in diagnosis of muscle loss were 36 mm, 72.0%, and 73.9%, respectively, for the male participants, and 34 mm, 72.2%, and 72.4%, respectively, for the female participants. In conclusion, the ultrasonography for thigh muscle might be a simple diagnostic method for sarcopenia.


Optimal programs of pathway control: dissecting the influence of pathway topology and feedback inhibition on pathway regulation.
Adjusting the capacity of metabolic pathways in response to rapidly changing environmental conditions is an important component of microbial adaptation strategies to stochastic environments. In this work, we use advanced dynamic optimization techniques combined with theoretical models to study which reactions in pathways are optimally targeted by regulatory interactions in order to minimize the regulatory effort that is required to adjust the flux through a complex metabolic network. Moreover, we analyze how constraints in the speed at which an organism can respond on a proteomic level influences these optimal targets of pathway control. We find that limitations in protein biosynthetic rates have a strong influence. With increasing protein biosynthetic rates the regulatory effort targeting the initial enzyme in a pathway is reduced while the regulatory effort in the terminal enzyme is increased. Studying the impact of allosteric regulation for different pathway topologies, we find that the presence of feedback inhibition by products of metabolic pathways allows organisms to reduce the regulatory effort that is required to control a metabolic pathway in all cases. In a linear pathway this even leads to the case where the sole transcriptional regulatory control of the terminal enzyme is sufficient to control flux through the entire pathway. We confirm the utilization of these pathway regulation strategies through the large-scale analysis of transcriptional regulation in several hundred prokaryotes. This work expands our knowledge about optimal programs of pathway control. Optimal targets of pathway control strongly depend on the speed at which proteins can be synthesized. Moreover, post-translational regulation such as allosteric regulation allows to strongly reduce the number of transcriptional regulatory interactions required to control a metabolic pathway across different pathway topologies.


Systematic analysis of cDNA microarray-based CGH.
cDNA microarray-based CGH (Microarray-CGH) is a useful technique for detecting genomic aberrations with a high resolution. However, the criteria for determining a genomic alteration have not been determined. We evaluated the genome-wide measurement of copy number of each gene in normal gastric and placenta tissues with both sex-matched, direct and sex-mismatched, indirect designs using 17K cDNA microarray. The results revealed the range of genomic copy number of normal tissues to be +/-0.3 of the log(2) ratio (gain >0.3, loss <-0.3) in the autosomal genes with direct and indirect designs. The copy number at a gene level from the X chromosomal genes using the direct and indirect sex-mismatched designs was +/-0.68 of the log(2) ratio (amplification >0.68, deletion <-0.68). In summary, the suggested method can be used as a guideline for analysis of genomic aberration using a Microarray-CGH in both direct and indirect designs.


Antibody-drug conjugates in glioblastoma therapy: the right drugs to the right cells.
Glioblastomas are high-grade brain tumours with a poor prognosis and, currently, few available therapeutic options. This lack of effective treatments has been linked to diverse factors, including target selection, tumour heterogeneity and poor penetrance of therapeutic agents through the blood-brain barrier and into tumours. Therapies using monoclonal antibodies, alone or linked to cytotoxic payloads, have proved beneficial for patients with different solid tumours; these approaches are currently being explored in patients with glioblastoma. In this Review, we summarise clinical data regarding antibody-drug conjugates (ADCs) against a variety of targets in glioblastoma, and compare the efficacy and toxicity of targeting EGFR with ADCs versus naked antibodies in order to illustrate key aspects of the use of ADCs in this malignancy. Finally, we discuss the complex challenges related to the biology and mutational changes of glioblastoma that can affect the use of ADC-based therapies in patients with this disease, and highlight potential strategies to improve efficacy.


The protective effects of silymarin against doxorubicin-induced cardiotoxicity and hepatotoxicity in rats.
Silymarin is a complex of five major compounds, and silibinin is the most biologically active component of the complex. The aim of this study was to investigate, evaluate and confirm the potential cardioprotective and hepatoprotective effects of administration of silymarin, rich in silibinin, at a dose of 60 mg/kg orally for a time-span of 12 days on doxorubicin induced toxicity in male Wistar rats. The in vivo model was used to explore whether silymarin could prevent damage of liver and heart tissue induced by doxorubicin administered every other day at dose of 1.66 mg/kg intraperitoneally for twelve days. In the study the change of body weight, ECG changes, biochemical parameters of oxidative stress, serum activity of alanine and aspartate transaminase, lactate dehydrogenase, creatine kinase and histological preparations of heart and liver samples of treated animals were examined. According to physiological, pharmacological, microscopic and biochemical results, we confirmed that at the examined dose, silymarin exhibits a protective influence on the heart and liver tissue against toxicity induced by doxorubicin.


Performance Characteristics of Galactomannan and β-d-Glucan in High-Risk Liver Transplant Recipients.
The utility of Aspergillus galactomannan (GM) and β-D-glucan (BG) in liver transplant recipients remains uncertain. As part of a randomized, double-blind trial of antifungal prophylaxis in liver transplant recipients at risk for invasive fungal infections (IFIs), GM and BG were assessed in 199 patients at baseline (enrollment) and weekly thereafter for the duration of study drug. Receiver operating characteristic (ROC) analysis was used to evaluate the accuracy of these for the diagnosis of IFIs. Overall, 46.4% of the patients at baseline had positive GM (index ≥ 0.5) and 89.6% had BG of 80 pg/mL or greater with BG level of 500 pg/mL or greater in 31.8%. Patients with invasive aspergillosis (IA) (3/3) had positive GM at baseline as did 45.5% of those without IA (P = 0.098); the area under the ROC curve for the diagnosis of IA was 0.77 (fair test, ie, good sensitivity but poor specificity). Using BG cutoff of 80 pg/mL or higher, 100% (12/12) of the patients with IFI had positive baseline BG and as did 88.9% (160/180) of those without IFI (P = 0.618); the area under the ROC curve for predicting IFIs was 0.56 (poor test). In multivariate analyses, GM positivity was associated with study site (P = 0.041), and BG positivity with renal replacement therapy (P = 0.05) and study site (P = 0.01). The GM and BG levels declined over time; positivity at subsequent time points was lower in comparison with baseline (P < 0.001). The GM and BG tests had significant center variability and limited accuracy for the diagnosis of IFIs in high-risk liver transplant recipients.


Evaluation of drug and sunscreen permeation via skin irradiated with UVA and UVB: comparisons of normal skin and chronologically aged skin.
Ultraviolet (UV) exposure is the predominant cause of skin aging. A systematic evaluation of drug skin permeation via photoaged skin is lacking. The aim of this work was to investigate whether UVA and UVB affect absorption by the skin of drugs and sunscreens, including tetracycline, quercetin, and oxybenzone. The dorsal skin of nude mice was subjected to UVA (24 and 39 J/cm(2)) or UVB (150, 200, and 250 mJ/cm(2)) irradiation. Levels of skin water loss, erythema, and sebum were evaluated, and histological examinations of COX-2 and claudin-1 expressions were carried out. Permeation of the permeants into and through the skin was determined in vitro using a Franz cell. In vivo skin uptake was also evaluated. Senescent skin (24 weeks old) was used for comparison. Wrinkling and scaling were significant signs of skin treated with UVA and UVB, respectively. The level of claudin-1, an indicator of tight junctions (TJs), was reduced by UVA and UVB irradiation. UVA enhanced tetracycline and quercetin skin deposition by 11- and 2-fold, respectively. A similar enhancement was shown for flux profiles. Surprisingly, a lower UVA dose revealed greater enhancement compared to the higher dose. The skin deposition and flux of tetracycline both decreased with UVB exposure. UVB also significantly reduced quercetin flux. The skin absorption behavior of chronologically aged skin approximated that of the UVA group, with photoaged skin showing higher enhancement. UV generally exhibited a negligible effect on modulating oxybenzone permeation. Skin disruption produced by UV does not necessarily result in enhanced skin absorption. It depends on the UV wavelength, irradiated energy, and physicochemical properties of the permeant. To the best of our knowledge, this is the first report establishing drug permeation profiles for UV-irradiated skin.


Posttranslational membrane attachment and dynamic fatty acylation of a neuronal growth cone protein, GAP-43.
Growth cones, the motile apparatus at the ends of elongating axons, are sites of extensive and dynamic membrane-cytoskeletal interaction and insertion of new membrane into the growing axon. One of the most abundant proteins in growth cone membranes is a protein designated GAP-43, whose synthesis increases dramatically in most neurons during periods of axon development or regeneration. We have begun to explore the role of GAP-43 in growth cone membrane functions by asking how the protein interacts with those membranes. Membrane-washing experiments indicate that mature GAP-43 is tightly bound to growth cone membranes, and partitioning of Triton X-114-solubilized GAP-43 between detergent-enriched and detergent-depleted phases indicates considerable hydrophobicity. The hydrophobic behavior of the protein is modulated by divalent cations, particularly zinc and calcium. In vivo labeling of GAP-43 in neonatal rat brain with [35S]methionine shows that GAP-43 is initially synthesized as a soluble protein that becomes attached to membranes posttranslationally. In tissue culture, both rat cerebral cortex cells and neuron-like PC12 cells actively incorporate [3H]palmitic acid into GAP-43. Isolated growth cones detached from their cell bodies also incorporate labeled fatty acid into GAP-43, suggesting active turnover of the fatty acid moieties on the mature protein. Hydrolysis of ester-like bonds with neutral hydroxylamine removes the bound fatty acid and exposes new thiol groups on GAP-43, suggesting that fatty acid is attached to the protein's only two cysteine residues, located in a short hydrophobic domain at the amino terminus. Modulation of the protein's hydrophobic behavior by divalent cations suggests that other domains, containing large numbers of negatively charged residues, might also contribute to GAP-43-membrane interactions. Our observations suggest a dynamic and reversible interaction of GAP-43 with growth cone membranes.


Integrin beta1 subunit controls mural cell adhesion, spreading, and blood vessel wall stability.
Growth, maturation, and integrity of the blood vessel network require extensive communication between the endothelial cells, which line the vascular lumen, and associated mural cells, namely vascular smooth muscle cells and pericytes. Pericytes extend long processes, make direct contact with the capillary endothelium, and promote vascular quiescence by suppressing angiogenic sprouting. Vascular smooth muscle cells are highly contractile, extracellular matrix-secreting cells that cover arteries and veins and provide them with mechanical stability and elasticity. In the damaged blood vessel wall, for example in atherosclerotic lesions, vascular smooth muscle cells lose their differentiated state and acquire a highly mitotic, so-called "synthetic" phenotype, which is thought to promote pathogenesis. Among other factors, extracellular matrix molecules and integrin family cell-matrix receptors may regulate this phenotypic transition. Here we show that the inactivation of the gene encoding the integrin beta1 subunit (Itgb1) with a Cre-loxP approach in mice leads to mural cell defects and postnatal lethality. Integrin beta1-deficient vascular smooth muscle cells display several hallmarks of the synthetic phenotype: Cell proliferation is enhanced, whereas differentiation and their ability to support blood vessels are compromised. Similarly, mutant pericytes are poorly spread but present in larger numbers. Our analysis of this mutant model shows that integrin beta1-mediated cell-matrix adhesion is a major determinant of the mural cell phenotype.


Combining DNA sequences and morphology in systematics: testing the validity of the dragonfly species Cordulegaster bilineata.
Morphological and molecular techniques are rarely combined when answering questions of taxonomic validity. In this study, we combine morphological techniques with DNA sequences to determine the validity of the dragonfly species Cordulegaster bilineata. The two dragonfly species C. bilineata and C. diastatops are very similar in size, body color, and morphological characters, and due to these similarities, the status of C. bilineata as a valid species is in question. In this study we compare morphological measurements of males and internal transcribed spacer 1 (ITS-1) sequences of rDNA between the two taxa. The hamule measurements (where copulation occurs) of males show little difference between the taxa in question, but the anal appendage measurements (where the male first contacts the female) show marked divergence between the two taxa. Cluster analysis with these anal appendage measurements correctly assigns almost all individuals measured into their respective taxon. PCR amplification products of ITS-1 display a approximately 50 bp size difference between C. bilineata (n = 4) and C. diastatops (n = 5) regardless of collection site. Sequence data for these amplifications show 51 bp missing in one locus in the ITS-1 of C. bilineatarelative to C. diastatops. A lone population of C. diastatops from Wisconsin has three individuals with ITS-1 products that match the size of both C. bilineata and C. diastatops. One individual from this population appears to yield two ITS-1 amplification products that match both C. bilineata and C. diastatops. Although this population may be evidence for hybridization between the two taxa, such hybridization is not necessarily sufficient to disqualify the validity of a separate species designation for C. bilineata. Morphology and ITS-1 sequences depict a high degree of divergence that is consistent with species-level differences.


Homo-psychologicus: Reactionary behavioural aspects of epidemics.
We formulate an in silico model of pathogen avoidance mechanism and investigate its impact on defensive behavioural measures (e.g., spontaneous social exclusions and distancing, crowd avoidance and voluntary vaccination adaptation). In particular, we use SIR(B)S (e.g., susceptible-infected-recovered with additional behavioural component) model to investigate the impact of homo-psychologicus aspects of epidemics. We focus on reactionary behavioural changes, which apply to both social distancing and voluntary vaccination participations. Our analyses reveal complex relationships between spontaneous and uncoordinated behavioural changes, the emergence of its contagion properties, and mitigation of infectious diseases. We find that the presence of effective behavioural changes can impede the persistence of disease. Furthermore, it was found that under perfect effective behavioural change, there are three regions in the response factor (e.g., imitation and/or reactionary) and behavioural scale factor (e.g., global/local) factors ρ-α behavioural space. Mainly, (1) disease is always endemic even in the presence of behavioural change, (2) behavioural-prevalence plasticity is observed and disease can sometimes be eradication, and (3) elimination of endemic disease under permanence of permanent behavioural change is achieved. These results suggest that preventive behavioural changes (e.g., non-pharmaceutical prophylactic measures, social distancing and exclusion, crowd avoidance) are influenced by individual differences in perception of risks and are a salient feature of epidemics. Additionally, these findings indicates that care needs to be taken when considering the effect of adaptive behavioural change in predicting the course of epidemics, and as well as the interpretation and development of the public health measures that account for spontaneous behavioural changes.


Elimination of a pressure ulcer with electrical stimulation--a case study.
Pressure ulcers, also called decubitus ulcers, are a common challenge of humanity and are exceptionally difficult to heal. They are wounds that are initiated by relatively short periods of pressure on the skin that blocks blood circulation causing the skin and underlying tissues to die, leading to an open wound. Pressure release can prevent further tissue degeneration, and some ulcers heal and disappear by themselves. However, many pressure ulcers never heal and continue to grow in diameter and depth. By one year, such unhealing ulcers are referred to as chronic ulcers. Chronic ulcers frequently jeopardize the life of the patient due to infections that become increasingly deep until they invade bones and the circulatory system. We report on a patient with a chronic pressure ulcer at his coccyx prominence. Fourteen months after the ulcer had appeared, a surface pulse electromagnetic force (PEMF) stimulator was applied over T7-T8, 45 cm cephalic to the ulcer, as part of a nerve stimulation study. Although the ulcer had continued to grow both in diameter and depth for 14 months and showed no signs of healing, within 6 days of applying the PEMF stimulator, the ulcer began to heal and was fully eliminated after 3 months. We concluded that the electrical stimulation induced the healing of the pressure ulcer. The ulcer elimination is quite surprising due to the exceptionally low electric field-force being generated by the stimulator at a distance of 45 cm.


Vancomycin resistant enterococci (VRE) in Swedish sewage sludge.
Antimicrobial resistance is a serious threat in veterinary medicine and human healthcare. Resistance genes can spread from animals, through the food-chain, and back to humans. Sewage sludge may act as the link back from humans to animals. The main aims of this study were to investigate the occurrence of vancomycin resistant enterococci (VRE) in treated sewage sludge, in a Swedish waste water treatment plant (WWTP), and to compare VRE isolates from sewage sludge with isolates from humans and chickens. During a four month long study, sewage sludge was collected weekly and cultured for VRE. The VRE isolates from sewage sludge were analysed and compared to each other and to human and chicken VRE isolates by biochemical typing (PhenePlate), PFGE and antibiograms. Biochemical typing (PhenePlate-FS) and pulsed field gel electrophoresis (PFGE) revealed prevalence of specific VRE strains in sewage sludge for up to 16 weeks. No connection was found between the VRE strains isolated from sludge, chickens and humans, indicating that human VRE did not originate from Swedish chicken. This study demonstrated widespread occurrence of VRE in sewage sludge in the studied WWTP. This implies a risk of antimicrobial resistance being spread to new farms and to the society via the environment if the sewage sludge is used on arable land.


Suicidal behaviour: an epidemiological and genetic study.
Psychiatric history, familial history of suicide attempts, and certain traumatic life events are important predictors of suicidal thoughts and behaviour. We examined the epidemiology and genetics of suicidality (i.e. reporting persistent suicidal thoughts or a plan or suicide attempt) in a large community-based sample of MZ and DZ twin pairs. Diagnostic telephone interviews were conducted in 1992-3 with twins from an Australian twin panel first surveyed in 1980-82 (N = 5995 respondents). Data were analysed using logistic regression models, taking into account twin pair zygosity and the history of suicidality in the respondent's co-twin. Lifetime prevalence of suicidal thoughts and attempts was remarkably constant across birth cohorts 1930-1964, and across gender. Major psychiatric correlates were history of major depression, panic disorder, social phobia in women, alcohol dependence and childhood conduct problems. Traumatic events involving assault (childhood sexual abuse, rape or physical assault) or status-loss (job loss, loss of property or home, divorce), and the personality trait neuroticism, were also significantly associated with suicide measures. Prevalence of serious suicide attempts varied as a function of religious affiliation. After controlling for these variables, however, history of suicide attempts or persistent thoughts in the respondent's co-twin remained a powerful predictor in MZ pairs (odds ratio = 3.9), but was not consistently predictive in DZ pairs. Overall, genetic factors accounted for approximately 45% of the variance in suicidal thoughts and behaviour (95% confidence interval 33-51%). Risk of persistent suicidal thoughts and suicide attempts is determined by a complex interplay of psychiatric history, neuroticism, traumatic life experiences, genetic vulnerability specific for suicidal behaviour and sociocultural risk or protective factors.


A gene-rich, transcriptionally active environment and the pre-deposition of repressive marks are predictive of susceptibility to KRAB/KAP1-mediated silencing.
KRAB-ZFPs (Krüppel-associated box domain-zinc finger proteins) are vertebrate-restricted transcriptional repressors encoded in the hundreds by the mouse and human genomes. They act via an essential cofactor, KAP1, which recruits effectors responsible for the formation of facultative heterochromatin. We have recently shown that KRAB/KAP1 can mediate long-range transcriptional repression through heterochromatin spreading, but also demonstrated that this process is at times countered by endogenous influences. To investigate this issue further we used an ectopic KRAB-based repressor. This system allowed us to tether KRAB/KAP1 to hundreds of euchromatic sites within genes, and to record its impact on gene expression. We then correlated this KRAB/KAP1-mediated transcriptional effect to pre-existing genomic and chromatin structures to identify specific characteristics making a gene susceptible to repression. We found that genes that were susceptible to KRAB/KAP1-mediated silencing carried higher levels of repressive histone marks both at the promoter and over the transcribed region than genes that were insensitive. In parallel, we found a high enrichment in euchromatic marks within both the close and more distant environment of these genes. Together, these data indicate that high levels of gene activity in the genomic environment and the pre-deposition of repressive histone marks within a gene increase its susceptibility to KRAB/KAP1-mediated repression.


Spatial framework of nine distinct local populations of the Japanese dormouse Glirulus japonicus based on matrilineal cytochrome b and patrilineal SRY gene sequences.
We previously revealed the presence of six genetically distinct matrilineal populations of the Japanese dormouse Glirulus japonicus in the distribution range of Honshu, Shikoku, and Kyushu islands. In this study, we extended this analysis using mitochondrial cytochrome b gene sequences (n = 96) and Y-chromosome-specific SRY gene sequences (n = 22) from individuals collected from Honshu, Shikoku, Kyushu, and Oki Dogo I. The cytochrome b sequence data allowed us to define precise geographic ranges of the six previously known and three newly found distinct matrilineal lineages: northeastern Honshu (I), east-central Honshu (II), west-central Honshu and the Kii Peninsula (III), the western part of Honshu (IV), Shikoku (V), westernmost Honshu and Kyushu (VI), the northern part of central Honshu (VII), the southern part of central Honshu (VIII), and Oki Dogo I. (IX). Our inference of geographic borders suggests that regions of lower and higher altitudes in the mountain systems played important roles in driving the hosting and separation of lineages, respectively. Six matrilineal lineages (I, II, V, VI, VIII, and XI) were shown to possess their own SRY haplotypes, while lineages III and IV shared one haplotype. These data together with our previous observation of nuclear ribosomal RNA gene variation indicate advanced populational subdivision in this species. It is thus evident that each of the populations, including those living at high latitudes and in limited geographic spaces, have survived for several million years. A specific ability to tolerate cold may have permitted G. japonicus to preserve anciently diverged lineages in each locality.


Practitioner review: Self-harm in adolescents.
Repeated self-harm in adolescents is common and associated with elevated psychopathology, risk of suicide, and demand for clinical services. Despite recent advances in the understanding and treatment of self-harm there have been few systematic reviews of the topic. The main aim of this article is to review randomised controlled trials (RCTs) reporting efficacy of specific pharmacological, social or psychological therapeutic interventions (TIs) in reducing self-harm repetition in adolescents presenting with self-harm. Data sources were identified by searching Medline, PsychINFO, EMBASE, and PubMed from the first available year to December 2010. RCTs comparing specific TIs versus treatment as usual or placebo in adolescents presenting with self-harm were included. Fourteen RCTs reported efficacy of psychological and social TIs in adolescents presenting with self-harm. No independently replicated RCTs have been identified reporting efficacy of TIs in self-harm reduction. Developmental Group Psychotherapy versus treatment as usual was associated with a reduction in repeated self-harm, however, this was not replicated in subsequent studies. Multisystemic Therapy (MST) versus psychiatric hospitalisation was associated with a reduction of suicidal attempts in a sample of adolescents with a range of psychiatric emergencies. However, analyses focusing only on the smaller subgroup of adolescents presenting with deliberate self-harm at the initial psychiatric emergency, did not indicate significant benefits of MST versus hospitalisation. Further research is urgently needed to develop TIs for treating self-harm in adolescents. MST has shown promise but needs to be evaluated in a sample of adolescents with self-harm; dialectic behavioural therapy and cognitive behavioural therapy for self-harm require RCTs to evaluate efficacy and effectiveness.


Close association between sequence polymorphism in the KIT gene and the roan coat color in horses.
The roan coat color in horses is controlled by a dominant allele that is lethal in the homozygous condition. Phenotypic similarities to some pigmentation disorders in human and mouse, combined with comparative mapping data, identified KIT, encoding the mast cell growth factor receptor, as a major candidate gene for the roan locus (Rn). Rn has previously been mapped to equine linkage group (LG) II. In this study, LGII was expanded with KIT and PDGFRA (platelet-derived growth factor receptor alpha) by use of RFLP and linkage analysis. Moreover, highly significant linkage disequilibrium between Rn and a KIT TaqI RFLP, representing a synonymous substitution in exon 19, was revealed. There was a strong KIT-Rn association in most breeds. Almost the complete KIT-encoding sequence was determined by sequence analysis of RT-PCR products. Comparison of horse KIT cDNA sequences, representing three different alleles (two different rn and one Rn), revealed five sequence polymorphisms and several mRNA splice variants, but none of these proved to be specifically associated with Rn. An insertion of a partial (79 bp) LINE1-element between exons 1 and 2, leading to a frameshift, represented about 30% of KIT transcripts in the Belgian roan horse used for the sequence analysis. However, an association between this L1 splice insertion and the roan phenotype was not verified when testing additional unrelated roan and non-roan horses from different breeds. The study strengthens the hypothesis that the roan coat color is controlled by KIT, but further analyses are needed to reveal the causative mutation(s).


Inhibition of spontaneous metastasis in a rat prostate cancer model by oral administration of modified citrus pectin.
Prostate cancer is the most common cancer diagnosed in U.S. men and remains incurable once it has metastasized. Many stages of the metastatic cascade involve cellular interactions mediated by cell surface components, such as carbohydrate-binding proteins, including galactoside-binding lectins (galectins). Modified citrus pectin (pH-modified), a soluble component of plant fiber derived from citrus fruit, has been shown to interfere with cell-cell interactions mediated by cell surface carbohydrate-binding galectin-3 molecules. The aim of this study was to determine whether modified citrus pectin, a complex polysaccharide rich in galactosyl residues, could inhibit spontaneous metastasis of prostate adenocarcinoma cells in the rat. The ability of modified citrus pectin to inhibit the adhesion of Dunning rat prostate cancer MAT-LyLu cells to rat endothelial cells was measured by 51Cr-labeling. Modified citrus pectin inhibition of MAT-LyLu cell anchorage-independent growth was measured by colony formation in agarose. The presence of galectin-3 in rat MAT-LyLu cells and human prostate carcinoma was demonstrated by immunoblotting and immunohistochemistry. One million MAT-LyLu cells were injected subcutaneously into the hind limb of male Copenhagen rats on day 0. Rats were given 0.0%, 0.01%, 0.1%, or 1.0% (wt/vol) modified citrus pectin continuously in their drinking water (from day 4 until necropsy on day 30). The number of MAT-LyLu tumor colonies in the lungs were counted. Compared with 15 or 16 control rats that had lung metastases on day 30, seven of 14 rats in the 0.1% and nine of 16 rats in the 1.0% modified citrus-pectin group had statistically significant (two-sided; P < .03 and P < .001, respectively) reductions in lung metastases. The lungs of the 1.0% modified citrus pectin-treated rats had significantly (two-sided; P < .05) fewer metastatic colonies than control groups (9 colonies +/- 4 [mean +/- SE] in the control group compared with 1 colony +/- 1 in the treated group). Modified citrus pectin had no effect on the growth of the primary tumors. In vitro, modified citrus pectin inhibited MAT-LyLu cell adhesion to rat endothelial cells in a time- and dose-dependent manner as well as their colony formation in semisolid medium. We present a novel therapy in which oral intake of modified citrus pectin acts as a potent inhibitor of spontaneous prostate carcinoma metastasis in the Copenhagen rat. Further investigations are warranted to determine the following: 1) the role of galectin-3 in normal and cancerous prostate tissues and 2) the ability of modified citrus pectin to inhibit human prostate metastasis in nude mice.


Enhanced regional terrestrial carbon uptake over Korea revealed by atmospheric CO2 measurements from 1999 to 2017.
Understanding changes in terrestrial carbon balance is important to improve our knowledge of the regional carbon cycle and climate change. However, evaluating regional changes in the terrestrial carbon balance is challenging due to the lack of surface flux measurements. This study reveals that the terrestrial carbon uptake over the Republic of Korea has been enhanced from 1999 to 2017 by analyzing long-term atmospheric CO2 concentration measurements at the Anmyeondo Station (36.53°N, 126.32°E) located in the western coast. The influence of terrestrial carbon flux on atmospheric CO2 concentrations (ΔCO2 ) is estimated from the difference of CO2 concentrations that were influenced by the land sector (through easterly winds) and the Yellow Sea sector (through westerly winds). We find a significant trend in ΔCO2 of -4.75 ppm per decade (p < .05) during the vegetation growing season (May through October), suggesting that the regional terrestrial carbon uptake has increased relative to the surrounding ocean areas. Combined analysis with satellite measured normalized difference vegetation index and gross primary production shows that the enhanced carbon uptake is associated with significant nationwide increases in vegetation and its production. Process-based terrestrial model and inverse model simulations estimate that regional terrestrial carbon uptake increases by up to 18.9 and 8.0 Tg C for the study period, accounting for 13.4% and 5.7% of the average annual domestic carbon emissions, respectively. Atmospheric chemical transport model simulations indicate that the enhanced terrestrial carbon sink is the primary reason for the observed ΔCO2 trend rather than anthropogenic emissions and atmospheric circulation changes. Our results highlight the fact that atmospheric CO2 measurements could open up the possibility of detecting regional changes in the terrestrial carbon cycle even where anthropogenic emissions are not negligible.


Mitosis detection in breast cancer histological images An ICPR 2012 contest.
In the framework of the Cognitive Microscope (MICO) project, we have set up a contest about mitosis detection in images of H and E stained slides of breast cancer for the conference ICPR 2012. Mitotic count is an important parameter for the prognosis of breast cancer. However, mitosis detection in digital histopathology is a challenging problem that needs a deeper study. Indeed, mitosis detection is difficult because mitosis are small objects with a large variety of shapes, and they can thus be easily confused with some other objects or artefacts present in the image. We added a further dimension to the contest by using two different slide scanners having different resolutions and producing red-green-blue (RGB) images, and a multi-spectral microscope producing images in 10 different spectral bands and 17 layers Z-stack. 17 teams participated in the study and the best team achieved a recall rate of 0.7 and precision of 0.89. Several studies on automatic tools to process digitized slides have been reported focusing mainly on nuclei or tubule detection. Mitosis detection is a challenging problem that has not yet been addressed well in the literature. Mitotic count is an important parameter in breast cancer grading as it gives an evaluation of the aggressiveness of the tumor. However, consistency, reproducibility and agreement on mitotic count for the same slide can vary largely among pathologists. An automatic tool for this task may help for reaching a better consistency, and at the same time reducing the burden of this demanding task for the pathologists. Professor Frιdιrique Capron team of the pathology department at Pitiι-Salpκtriθre Hospital in Paris, France, has selected a set of five slides of breast cancer. The slides are stained with H and E. They have been scanned by three different equipments: Aperio ScanScope XT slide scanner, Hamamatsu NanoZoomer 2.0-HT slide scanner and 10 bands multispectral microscope. The data set is made up of 50 high power fields (HPF) coming from 5 different slides scanned at ×40 magnification. There are 10 HPFs/slide. The pathologist has annotated all the mitotic cells manually. A HPF has a size of 512 μm × 512 μm (that is an area of 0.262 mm (2) , which is a surface equivalent to that of a microscope field diameter of 0.58 mm. These 50 HPFs contain a total of 326 mitotic cells on images of both scanners, and 322 mitotic cells on the multispectral microscope. Up to 129 teams have registered to the contest. However, only 17 teams submitted their detection of mitotic cells. The performance of the best team is very promising, with F-measure as high as 0.78. However, the database we provided is by far too small for a good assessment of reliability and robustness of the proposed algorithms. Mitotic count is an important criterion in the grading of many types of cancers, however, very little research has been made on automatic mitotic cell detection, mainly because of a lack of available data. A main objective of this contest was to propose a database of mitotic cells on digitized breast cancer histopathology slides to initiate works on automated mitotic cell detection. In the future, we would like to extend this database to have much more images from different patients and also for different types of cancers. In addition, mitotic cells should be annotated by several pathologists to reflect the partial agreement among them.


Synthesis and photophysical properties of a bichromophoric system hosting a disaccharide spacer.
The synthesis of an efficient energy donor-acceptor system is reported, together with its photophysical properties. The bichromophoric species has been conceived to show potentialities for biological applications since a biocompatible disaccharide spacer, constituted of d-galactose and d-glucose derivatives, was used in compound 12 to connect two BODIPY units with different absorption/emission properties. The luminescence spectrum in acetonitrile of 12 shows an intense fluorescence band with a maximum at about 770 nm that is almost identical to that of the lowest-energy BODIPY, regardless of the excitation wavelength used. The quantum yield is 0.2 with an excited state lifetime of 2.5 ns. Excitation and ultrafast transient absorption spectroscopy demonstrates that a very efficient energy transfer takes place in 12 from the highest-energy lying BODIPY subunit to the lowest-energy emissive BODIPY moiety, with a time constant of about 31 ps. Noteworthily, the emission of 12 falls in the near infrared window, suitable for potential biological applications.


Short-course therapy for catheter-associated Staphylococcus aureus bacteremia.
To determine the efficacy of "short-course" therapy (less than 17 days) for Staphylococcus aureus catheter-associated bacteremia, 13 patients were prospectively followed up for at least three months after completion of therapy. A single patient relapsed after 28 days with endocarditis. No clinical or microbiological predictors of relapse could be identified, and coexistent medical conditions associated with some degree of immunosuppression did not appear to predispose to relapse. The results of this study and a review of the literature indicate that short-course therapy for uncomplicated S aureus catheter-associated bacteremia has a relapse rate of only 5% to 10% and, therefore, is reasonable therapy for this condition. The majority of relapses are endocarditis and occur within ten weeks after completion of therapy. Close follow-up during this period is essential.


Bioactivity of cis and dicis isomers of vitamin A esters.
In order to declare the accurate vitamin A effectiveness of foodstuffs and synthetic vitamin A preparations, the biological activity of the individual isomers has to be known. Total activity is expressed as all-trans vitamin A equivalents which are calculated by multiplication of analytical content of each isomer with the corresponding biopotency factor. The biological activity of four cis and three or four dicis isomers of vitamin A acetate and palmitate was determined by means of rat vaginal smear assays. The purity of the compounds, the content and isomerisation in stock solutions were checked by HPLC methods prior to their dilution and application to the experimental animals. Based on analytical contents the following activities were found for vitamin A acetate isomers: all-trans 1.00, 13-cis 0.76, 11-cis 0.31, 9-cis 0.19, 7-cis 0.18, 9,13-dicis 0.16, 11,13-dicis 0.18, 9,11-dicis approx. 0.03. The corresponding values for vitamin A palmitate were: all-trans 1.00, 13-cis 0.73, 11-cis 0.34, 9-cis 0.19, 7-cis 0.14, 9,13-dicis 0.21, 11,13-dicis 0.20, 9,11-dicis approx. 0.04, 7,11-dicis approx. 0.12. The vitamin A activity of the cis isomers ranged from about 0.75 for 13-cis to about 0.05 for 9,11-dicis. For the cis series, a biopotency relationship depending on the distance between the beta-ionone ring system and the cis configuration could be observed.


Insulin-like growth factors and subsequent risk of mortality in the United States.
Although numerous studies have explored the relation of insulin-like growth factor (IGF)-I and IGF-binding protein (BP) 3 with cancer and cardiovascular disease, only two previous studies are known to have looked at the association of IGF-I and IGF-BP3 with risk of mortality. The objective of this US study was to examine the risk of all-cause, heart disease, and cancer mortality associated with IGF-I and IGF-BP3 levels using data from the Third National Health and Nutrition Examination Survey (NHANES III) and NHANES III Mortality Study (n = 6,061) (1988-2000). The authors constructed proportional hazards models with age as the time scale to determine the association of baseline IGF-I and IGF-BP3 levels with subsequent mortality. After adjustment for baseline measures, there was no increased risk of all-cause, heart disease, or cancer mortality for the lower quartiles of IGF-I compared with the highest quartile. The adjusted relative hazard of all-cause mortality for the lowest quartile of IGF-BP3 compared with the highest quartile was 1.57 (95% confidence interval: 0.98, 2.52), and the trend for risk was significant (p = 0.0364), but there was no increased risk of heart disease or cancer mortality. Results suggest that the association of IGF-I and IGF-BP3 with mortality may differ from associations with incidence of disease.


p53 gene gets altered by various mechanisms: studies in childhood sarcomas and retinoblastoma.
Somatic and constitutional mutation screening of p53 in childhood sarcomas and retinoblastoma was investigated by a multitechnical approach to evaluate its role in the development/progression by somatic mutation events and/or genetic predisposition. The studies were carried out on a cohort of 100 sarcoma cases, i.e. Ewing's sarcoma (n=44), osteosarcoma (n=36), and rhabdomyosarcoma (n= 20), and on 50 retinoblastoma (Rb) cases. Constitutional allelic deletion was found by FISH in 4% of sarcoma cases. Overall, 20% of sarcoma tumors showed p53 rearrangement by PCR/SSCP and Southern blot. Allelic deletion of p53 was detected in 78% of sarcoma and 55% of Rb tumors. p53 protein expression was detected by immunohistochemistry in 20% of sarcoma tumors. This study for the first time provided evidence of p53 alteration through allelic deletion that are common primary somatic mutation events which occur irrespective of grade and stage and are hence probably associated with an early phase of tumorigenesis and/or tumor progression. The studies also explored the occurrence of de novo constitutional deletion of p53 in sporadic childhood sarcomas. This study in retinoblastoma provided evidence for the synergistic role of RB1 and p53, probably essential for the full-blown development of malignancy.


Long-term risk of malignancy after treatment of inflammatory bowel disease with azathioprine.
Data from renal transplant and rheumatoid arthritis patients suggest that there is an increased risk of malignancy after treatment with azathioprine. Whether this is true for patients with inflammatory bowel disease remains uncertain. A retrospective review of clinical notes was performed. Azathioprine was given to 626 of 2204 patients (855 with Crohn's disease and 1349 with ulcerative colitis). The mean total duration of azathioprine use was 27 months. The mean follow-up from diagnosis was 13.7 years and the mean follow-up from the start of azathioprine treatment was 6.9 years. Thirty-one cancers were observed in 30 patients treated with azathioprine (4.5%) and 77 cancers were observed in 70 patients not treated with azathioprine (4.5%; P=N.S.). Logistic regression analysis (including in the model the age, sex, diagnosis and extent of disease) showed that treatment with azathioprine did not significantly affect the risk of the development of cancer. Eight patients had lymphoma; three had been given azathioprine (P=N.S.). For patients with ulcerative colitis, the number of colorectal cancers (including high-grade dysplasia) in patients given azathioprine was eight of 355 (2.2%), compared with 28 of 994 (2.8%) for patients not given azathioprine (P=N.S.). The cumulative risk of colorectal cancer or dysplasia/dysplasia-associated lesion or mass (adjusted to exclude post-colectomy patients) after 10, 20, 30 and 40 years of ulcerative colitis was 0.4%, 1.3%, 9%and 15.5%, respectively. No increased risk of cancer diagnosis following azathioprine treatment was observed.


Cholesterol granuloma of the jaws: report of two cases.
Cholesterol granuloma (CG) is a foreign body reaction to the deposition and inadequate drainage of cholesterol crystals that are frequently found in association with chronic middle ear diseases but an uncommon entity in maxillary sinuses. The clinical symptoms are non-specific and depend on the localization and extent in each individual case. Bone erosion may be seen in cholesterol granuloma showing expansive growth. The differential diagnosis of CG includes cysts, mucocoeles and neoplasms. Radiological and histopathological findings are essential for diagnosis of CG. In this report it is aimed to introduce two new CG cases with its clinical, histopathological and radiological findings


[Pulmonary prognosis in preterm infants].
The increased survival of preterm infants poses the challenge of dealing with a wide range of chronic pulmonary diseases, including bronchopulmonary dysplasia, Wilson-Mikity syndrome and recurrent wheezing. This article reviews the pulmonary clinical and functional prognosis of preterm newborns in infancy and adolescence. MEDLINE search for articles published between 1970 and 2004 that focused on lung growth and function of preterm infants, besides a clinical follow-up of this group. Prenatal and postnatal events, such as placental insufficiency, tobacco exposure, infections, oxygen and mechanical ventilation, have an important effect on lung development and can lead to chronic lung diseases, of which bronchopulmonary dysplasia is the most severe complication. However, significant loss of lung function occurs in preterm infants who do not fulfill the criteria for bronchopulmonary dysplasia, and even in those who did not have significant respiratory disease during the neonatal period. The impact of prematurity on the respiratory system of these patients is usually underestimated. Clinically, preterm infants have an increased incidence of pneumonia and bronchiolitis, hospital readmissions due to respiratory diseases, chronic cough and wheezing and bronchial hyperresponsiveness. In adolescence, there is a tendency for normalization of the lung function, but they persist with reduced flows, lower exercise tolerance and bronchial hyperresponsiveness. Prematurity, the events that cause it and the interventions that follow it permanently change the development of the respiratory system. Studies are necessary to clarify the effect of each of these perinatal insults on the development of the respiratory system.


CD4 and the immunoglobulin superfamily.
The CD4 membrane glycoprotein was one of the first cell surface antigens to be identified using monoclonal antibodies. It was shown to have a central role in the control of the recognition of foreign proteins by T lymphocytes and later as a receptor for the human immunodeficiency virus (HIV). The analysis of the amino acid sequence of CD4 showed that the extracellular region comprised four regions with sequence similarities to immunoglobulin domains. The structure of domains 3 and 4 of CD4 has been determined by X-ray crystallography and, like domains 1 and 2 previously determined, these have typical immunoglobulin-like folds. The results are discussed with respect to the identification of other domains with immunoglobulin-like folds from amino acid sequence data, and the evolution of CD4.


Quantitative and Statistical Study of the Dynamics of Clathrin-Dependent and -Independent Endocytosis Reveal a Differential Role of EndophilinA2.
Eukaryotic cells internalize cargos specifically through clathrin-mediated endocytosis (CME) or clathrin-independent endocytosis (CIE). EndophilinA2 was shown as preferentially implicated in CIE, although initially involved in CME. Here, we investigated the native interplay of endophilinA2 and dynamin2 during CME as compared to CIE. We developed an unbiased integrative approach based on genome engineering, robust tracking methodology, and advanced analytics. We statistically identified CME and CIE subpopulations corresponding to abortive, active, and static endocytic events. Depletion of dynamin2 strongly affected active CME and CIE events, whereas the absence of endophilinA2 impacted only CIE. Accordingly, we demonstrated that endophilinA2 is needed for dynamin2 recruitment during CIE, but not in CME. Despite these differences, endophilinA2 and dynamin2 acted at the latest stage of endocytosis within a similar stoichiometry in both mechanisms. Thus, we propose a conserved function of dynamin2 and endophilinA2 in vesicle scission, but a differential regulation of their recruitment during CME and CIE.


Nuclear magnetic resonance and calorimetric study of the structure, dynamics, and phase behavior of uranyl ion/dipalmitoylphosphatidylcholine complexes.
The interaction of UO2(2+) with dipalmitoylphosphatidylcholine (DPPC) has been studied as a function of temperature and composition using nuclear magnetic resonance (NMR) spectroscopy, differential scanning calorimetry (DSC), and monolayer studies. Computer simulations of the 31P-NMR powder spectra of DPPC dispersions in the presence of various concentrations of UO2(2+) are consistent with the binding stoichiometry of [UO2(2+)]/[DPPC] = 1:4 at [UO2(2+)]/[DPPC] less than 0.3. This complex undergoes a phase transition to the liquid crystalline phase at T'm = 50 +/- 3 degrees C with a breadth delta T'm = 7 +/- 3 degrees C. This broad transition gradually disappears at higher UO2(2+) concentrations, suggesting the presence of yet another UO2(2+)/DPPC complex (or complexes) whose NMR spectra are indistinguishable from those of the 1:4 UO2(2+)/DPPC species. The temperature-dependent 13C powder spectra of 2(1-13C) DPPC dispersions in the presence of 1.2 mol ratio of UO2(2+) show that this higher order complex (complexes) also undergoes a phase transition to the liquid crystalline state at T'm +/- = 58 +/- 3 degrees C with a breadth delta T"m = 15 +/- 5 degrees C. The NMR spectra indicate that exchange among these various UO2(2+)/DPPC complexes is slow. In addition, computer simulations of the 31P-, 13C-, and 2H-NMR powder spectra show that axial diffusion of the DPPC molecules about their long axes is quenched by addition of UO2(2+) and acyl chain isomerization is the dominant motional mode. The isomerization is best described as two-site hopping of the greater than C-D bond at a rate of approximately 10(6) s-1, a motional mode which is expected for a kink diffusion.


Potent anticoagulants are associated with a higher all-cause mortality rate after hip and knee arthroplasty.
Anticoagulation for thromboprophylaxis after THA and TKA has not been confirmed to diminish all-cause mortality. We determined whether the incidence of all-cause mortality and pulmonary embolism in patients undergoing total joint arthroplasty differs with currently used thromboprophylaxis protocols. We reviewed articles published from 1998 to 2007 that included 6-week or 3-month incidence of all-cause mortality and symptomatic, nonfatal pulmonary embolism. Twenty studies included reported 15,839 patients receiving low-molecular-weight heparin, ximelagatran, fondaparinux, or rivaroxaban (Group A); 7193 receiving regional anesthesia, pneumatic compression, and aspirin (Group B); and 5006 receiving warfarin (Group C). All-cause mortality was higher in Group A than in Group B (0.41% versus 0.19%) and the incidence of clinical nonfatal pulmonary embolus was higher in Group A than in Group B (0.60% versus 0.35%). The incidences of all-cause mortality and nonfatal pulmonary embolism in Group C were similar to those in Group A (0.4 and 0.52, respectively). Clinical pulmonary embolus occurs despite the use of anticoagulants. Group A anticoagulants were associated with the highest all-cause mortality of the three modalities studied.


Normal pressure hydrocephalus due to an iatrogenic giant lumbar pseudomeningocele after posterior lumbar interbody fusion: illustrative case.
Iatrogenic pseudomeningocele incidence after lumbar surgery is 0.068%-2%, and most lumbar pseudomeningoceles are smaller than 5 cm; however, in rare cases, "giant" pseudomeningoceles greater than 8 cm in size may develop. Normal pressure hydrocephalus (NPH) is another rare condition in which the ventricles expand despite the presence of normal intracranial pressure. To date, pseudomeningocele associated with NPH has not been reported. An 80-year-old woman underwent L3-5 laminectomy and posterior lumbar interbody fusion, and her symptoms improved after surgery. However, dementia appeared 1 month after surgery. Repeated brain computed tomography showed ventricular enlargement, and lumbar magnetic resonance imaging showed a long pseudomeningocele in the subcutaneous tissues at the L4 level. Here, the authors report a rare case of an iatrogenic giant pseudomeningocele accompanied by NPH after lumbar surgery. The symptoms of NPH in the present case occurred after spinal surgery and recovered after dural repair surgery, indicating that the changes in cerebrospinal fluid circulation and/or pressure due to pseudomeningoceles may cause NPH. The prevention of dural tears through precise surgical technique and primary repair of dural tears are the best approaches to prevent pseudomeningocele incidence and subsequent events.


Rescue high frequency ventilation for congenital diaphragmatic hernia.
High frequency jet (HFJV) and oscillatory (HFOV) ventilation were used to rescue newborns with congenital diaphragmatic hernia (CDH), who failed conventional mechanical ventilation (CV). Changes in ventilator settings and pulmonary gas exchange were evaluated following transition to high frequency ventilation (HFV). Records of patients with CDH rescued with HFV prior to surgical intervention between 2006 and 2015 were reviewed. Mean airway pressure (Pāw) and arterial blood gases during CV and those obtained within the first hour of HFV were compared. A composite repeated measure analysis was performed to evaluate longitudinal and intergroup variances. Twenty-seven patients were rescued from CV, 16 by HFJV and 11 by HFOV. The two groups had similar gestational ages and birth weights. Prior to HFV, both groups had similar Pāw, PaCO2, FiO2 and PaO2. HFV was associated with a significant improvement in ventilation, and the rate of decrease of PaCO2 was no different between groups. There was a significantly higher increase in Pāw increase with HFOV compared to HFJV. In newborns with CDH rescued with HFV, ventilation improved but Pāw was significantly lower in patients supported with HFJV compared to HFOV.


Learning Inverse Statics Models Efficiently With Symmetry-Based Exploration.
Learning (inverse) kinematics and dynamics models of dexterous robots for the entire action or observation space is challenging and costly. Sampling the entire space is usually intractable in terms of time, tear, and wear. We propose an efficient approach to learn inverse statics models-primarily for gravity compensation-by exploring only a small part of the configuration space and exploiting the symmetry properties of the inverse statics mapping. In particular, there exist symmetric configurations that require the same absolute motor torques to be maintained. We show that those symmetric configurations can be discovered, the functional relations between them can be successfully learned and exploited to generate multiple training samples from one sampled configuration-torque pair. This strategy drastically reduces the number of samples required for learning inverse statics models. Moreover, we demonstrate that exploiting symmetries for learning inverse statics models is a generally applicable strategy for online and offline learning algorithms. We exemplify this by two different learning approaches. First, we modify the Direction Sampling approach for learning inverse statics models online, in a plain exploratory fashion, from scratch and without using a closed-loop controller. Second, we show that inverse statics mappings can be efficiently learned offline utilizing lattice sampling. Results for a 2R planar robot and a 3R simplified human arm demonstrate that their inverse statics mappings can be learned successfully for the entire configuration space. Furthermore, we demonstrate that the number of samples required for learning inverse statics mappings for 2R and 3R manipulators can be reduced at least by factors of approximately 8 and 16, respectively-depending on the number of discovered symmetries.


Cell death regulation by the mammalian IAP antagonist Diablo/Smac.
In Drosophila, the genetic locus 75CI1,2 is essential for all developmental cell death. Within this region are the genes for three pro-death proteins, Grim, Reaper and HID. These proteins are transcriptionally regulated and their expression tightly associated with cell death in the developing fly embryo. When ectopically expressed in the retina, Grim, Reaper and HID cause apoptosis and eye ablation. They have a short region of similarity at their N-termini through which they can interact with inhibitor of apoptosis (IAP) proteins, and it is by antagonising IAP inhibition of caspases that Grim, Reaper and HID promote cell death. The observation that Grim, Reaper and HID can interact with mammalian IAPs and induce apoptosis in mammalian cells suggested that mammalian IAP antagonists might also exist. Diablo/Smac, identified six years after the first description of a Drosophila IAP antagonist, is the only mammalian protein identified to date that is clearly functionally related to the Drosophila proteins. Since its discovery, there have been numerous studies investigating how Diablo/Smac interacts with IAPs and promotes cell death. Here we review what is currently known about Diablo/Smac and speculate on other mammalian IAP antagonists.


Reproductive biology of the spotback skate Atlantoraja castelnaui in the south-west Atlantic Ocean.
This study provides information on the reproduction of spotback skate Atlantoraja castelnaui. A total of 232 individuals (119 females and 113 males) were obtained from surveys carried out between 2003 and 2006, from the south-west Atlantic Ocean, between 34 and 42° S and <50 m deep; another 514 specimens (241 females and 273 males) were obtained between 2005 and 2007 from commercial fishery operations carried out in the same area and landings in the port of Mar del Plata, Argentina. Males ranged from 185 to 1250 mm total length (L(T) ) and females from 243 to 1368 mm L(T) . Length at maturity was estimated to be 980 mm for males and 1089 mm L(T) for females. Lack of variation of testis mass together with the continuous production of mature spermatocyst and spermatozoa in deferent ducts suggested that males can reproduce throughout the year. Females reproduced year-round with peaks of reproductive activity an integral part of a continuous cycle. This conclusion is corroborated by the seasonal variation of ovaries, oviducal gland and the occurrence of females with eggs in the uterus throughout the year. Results from this study indicate that A. castelnaui is very susceptible to fishery pressure.


Hydrogen-hydrogen bonding in planar biphenyl, predicted by atoms-in-molecules theory, does not exist.
Based on an Atoms-in-Molecules (AIM) analysis, Matta et al. recently claimed evidence for the existence of hydrogen-hydrogen bonding between ortho-hydrogen atoms, pointing towards each other from adjacent phenyl groups in planar biphenyl. This AIM result is opposed to the classical view that nonbonded steric repulsion between the ortho-hydrogen atoms is responsible for the higher energy of the planar as compared to the twisted geometry of biphenyl. In the present work, we address the question if hydrogen-hydrogen bonding in biphenyl exists, as suggested by AIM, or not. To this end, we have analyzed the potential energy surface for internal rotation of biphenyl in terms of two interacting phenyl radicals using density functional theory (DFT) at BP86/TZ2P. A detailed analysis of the bonding mechanism and a quantitative bond energy decomposition in the framework of Kohn-Sham DFT show that Pauli (or overlap) repulsion, mainly between C(ortho)--H(ortho) phenyl MOs, prevents biphenyl from being planar and forces it to adopt a twisted equilibrium geometry. Furthermore, a derivative of biphenyl in which all four ortho-hydrogen atoms have been removed does adopt a planar equilibrium geometry. Thus, our results confirm the classical view of steric repulsion between ortho-hydrogen atoms in biphenyl and they falsify the hypothesis of hydrogen-hydrogen bonding.


Risk for oral cancer associated to smoking, smokeless and oral dip products.
Oral cancer is one of the most common life threatening diseases in India. Tobacco and alcohol are considered to be the most risk factors for oral cancer. This study was conducted to investigate the association of tobacco and poly-ingredient oral dip products with oral cancer. A case-control study of 350 cases and 350 controls, over a period of 19 months, between February 2005 and September 2006 was carried out in Pune, India. The self-reported information about the consumption of tobacco, poly-ingredient oral dip products, alcohol, dietary habits and demographic status were collected by a researcher made questionnaire. Univariate and multivariate analysis were used to identify the risk of substances abuse. The frequency of smoking, smokeless and oral dip products in cases were significantly higher than controls (P < 0.0001). Among smoking types, bidi (P < 0.0001, OR = 4.1 95% CI = 2.4 - 6.9), of smokeless types, chewing tobacco (P < 0.0001, OR = 8.3, 95% CI = 5.4 - 13.0) and mishiri (P < 0.0001, OR = 3.3, 95% CI =2.1 - 5.4), and of oral dip products, consumption of gutkha (P < 0.0001, OR = 12.8, 95% CI =7.0 - 23.7) and supari (P < 0.0001, OR = 6.6, 95% CI =3.0 - 14.8) indicated strong association with oral cancer upon adjustment. This study provides strong evidence that gutkha, supari -areca nut- chewing tobacco (tobacco flakes), bidi smoking and mishiri (tobacco powder, which applied as a tooth and gum cleaner) are independent risk for oral cancer.


The optimal dosage and window of opportunity to maintain mitochondrial homeostasis following traumatic brain injury using the uncoupler FCCP.
Experimental traumatic brain injury (TBI) leads to a rapid and extensive necrosis at the primary site of injury that appears to be driven in part by significant mitochondrial dysfunction. The present study is based on the hypothesis that TBI-induced, aberrant glutamate release increases mitochondrial Ca(2+) cycling/overload ultimately leading to mitochondrial damage. Previous work from our laboratory demonstrates that mitochondrial uncoupling during the acute phases of TBI-induced excitotoxicity can reduce mitochondrial Ca(2+) uptake (cycling), ROS production and mitochondrial damage resulting in neuroprotection and improved behavioral outcome. The current study was designed to determine the optimal dosage and therapeutic window of opportunity for the potent mitochondrial uncoupler FCCP following moderate TBI. For this study, we used young adult male Sprague-Dawley rats (300-350 g); either sham-operated or moderately (1.5 mm) injured using the controlled cortical impactor (CCI) model of TBI. In the first set of studies animals were injected with either vehicle (100% DMSO) or different concentrations of FCCP (0.5, 1, 2.5 and 5 mg/kg in 100% DMSO) intraperitoneally at 5 min post-injury; tested behaviorally at 10 days and cortical sparing assessed at 18 days post-injury. The results demonstrate that of all the dosages tested, 2.5 mg/kg rendered the maximum improvement in behavioral outcomes and tissue spared. Using this optimal dose (2.5 mg/kg) and time point for intervention (5 min post-injury), we assessed mitochondrial bioenergetics and mitochondrial structural integrity 24 h post-injury. Furthermore, using this dosage we assessed mitochondrial bioenergetics and Ca(2+) loading at 3 and 6 h post-injury to further verify our target mechanism and establish these assessments as a valid endpoint to use as a means to determine the therapeutic window of FCCP. To begin to address the window of opportunity for maintaining mitochondrial homeostasis, the optimal dose of FCCP was then administered at 5 min, 3, 6, or 24 h post-injury and several parameters of mitochondrial function were used as outcome measures. The results demonstrate that a prolonged window of opportunity exists for targeting mitochondrial dysfunction using uncouplers following TBI and give insight into the cellular pathology associated with TBI.


The phosphatidylserine receptor has essential functions during embryogenesis but not in apoptotic cell removal.
Phagocytosis of apoptotic cells is fundamental to animal development, immune function and cellular homeostasis. The phosphatidylserine receptor (Ptdsr) on phagocytes has been implicated in the recognition and engulfment of apoptotic cells and in anti-inflammatory signaling. To determine the biological function of the phosphatidylserine receptor in vivo, we inactivated the Ptdsr gene in the mouse. Ablation of Ptdsr function in mice causes perinatal lethality, growth retardation and a delay in terminal differentiation of the kidney, intestine, liver and lungs during embryogenesis. Moreover, eye development can be severely disturbed, ranging from defects in retinal differentiation to complete unilateral or bilateral absence of eyes. Ptdsr -/- mice with anophthalmia develop novel lesions, with induction of ectopic retinal-pigmented epithelium in nasal cavities. A comprehensive investigation of apoptotic cell clearance in vivo and in vitro demonstrated that engulfment of apoptotic cells was normal in Ptdsr knockout mice, but Ptdsr-deficient macrophages were impaired in pro- and anti-inflammatory cytokine signaling after stimulation with apoptotic cells or with lipopolysaccharide. Ptdsr is essential for the development and differentiation of multiple organs during embryogenesis but not for apoptotic cell removal. Ptdsr may thus have a novel, unexpected developmental function as an important differentiation-promoting gene. Moreover, Ptdsr is not required for apoptotic cell clearance by macrophages but seems to be necessary for the regulation of macrophage cytokine responses. These results clearly contradict the current view that the phosphatidylserine receptor primarily functions in apoptotic cell clearance.


Transport kinetics of amino acids across the resting human leg.
Flux rates of amino acids were measured across the leg after an overnight fast in resting human volunteers. A balanced amino acid solution was, after a primed infusion, continuously infused for 2 h at each of three step-wise and increasing rates corresponding to 8.3, 16.7, 33.2 mg N/kg per h that were equivalent to 0.2, 0.4, 0.8 g N/kg per d. Flux of amino acids across the leg was compared with the flux of glucose, glycerol, lactate, free fatty acids, and oxygen. The size of the muscular tissue pool of amino acids was measured. Whole body amino acid oxidation was estimated by means of the continuous infusion of a 14C-labeled mixture of amino acids. Arterial steady state levels were obtained for most amino acids within 30 to 45 min after the primed constant infusion. Leg flux of amino acids switched from a net efflux after an overnight fast to a balanced flux between infusion rates corresponding to 0.2-0.4 g N/kg per d. At 0.8 g N/kg per d essentially all amino acids showed uptake. The infusion of amino acids stimulated leg uptake of glucose and lactate production and decreased FFA release. Oxygen uptake and leg blood flow increased significantly with increased infusion of amino acids. There was significant variability in transport rate among individual amino acids. Branched chain amino acids showed rapid transport and methionine slow transport rate. Only small changes in the muscle tissue concentration of certain amino acids were registered after 6 h of amino acid infusion despite uptake for several hours. When amino acids were infused at a rate corresponding to 0.8 g N/kg per d, the leg uptake of amino acids was 6% and the simultaneous whole body oxidation of infused amino acids was approximately 10%. Net uptake of leucine across the leg per hour was 62% of the muscle pool of free leucine when amino acids were infused at a rate corresponding to 0.4 g N/kg per d. Multiple regression analysis showed that the arterial concentration of an amino acid was the most important factor for uptake, more so than insulin concentration and blood flow. It is concluded that leg exchange of amino acids is large enough to rapidly change the pool size of the amino acids in skeletal muscle, if not counter-regulated by changes in rates of protein synthesis and degradation. Estimates of the capacity for protein synthesis and transfer RNA acceptor sites in muscles agree in order of magnitude with the net uptake of amino acids at high infusion rates of amino acids. Therefore, measurements of the balance of tyrosine, phenylalanine, and particularly methionine at steady state may reflect net balance of proteins across skeletal muscles even in short-time experiments.


Prevalence of remission and its effect on damage and quality of life in Chinese patients with systemic lupus erythematosus.
To study the prevalence of remission and its effect on damage and quality of life (QOL) in Chinese patients with systemic lupus erythematosus (SLE). Patients who fulfilled ≥4 American College of Rheumatology criteria for SLE were identified. Their remission status at last clinic visits was determined by the European consensus criteria (complete/clinical remission ± immunosuppressive drugs). The increase in SLE damage index (SDI) in the preceding 5 years was compared between patients who were and were not in remission for ≥5 years. QOL of patients as assessed by the validated Chinese version of the Medical Outcomes Study Short-Form-36 (SF36) and the LupusPRO was also compared between the remission and non-remission groups by statistical analysis. 769 SLE patients were studied (92% women; age: 46.4±14.6 years; SLE duration: 12.6±8.1 years). At last visit, clinical remission was present in 259 (33.7%) patients and complete remission was present in 280 (36.4%) patients. Clinical and complete remissions for ≥5 years were achieved in 64 (8.3%) and 129 (16.8%) of the patients, respectively. Patients remitted for ≥5 years were older, and had significantly lower prevalence of renal involvement, leucopenia or thrombocytopaenia. Fifty-three (6.9%) patients in remission ≥5 years were taken off all medications, including hydroxychloroquine (HCQ) (drug-free). Patients who remitted for ≥5 years but off-therapy (except HCQ) had significantly less SDI increment than those who did not remit (0.17±0.53 vs 0.67±1.10; p<0.001). Among 453 patients who had QOL assessment, remission for ≥5 years was associated with significantly higher SF36 and the total health-related scores of the LupusPRO. Durable remission can be achieved in a quarter of patients with SLE. Patients with remission for ≥5 years have significantly less damage accrual and better QOL. Prolonged remission is an appropriate criterion for outcome assessment in SLE.


A bayesian approach to deformed pattern matching of iris images.
We describe a general probabilistic framework for matching patterns that experience in-plane nonlinear deformations, such as iris patterns. Given a pair of images, we derive a maximum a posteriori probability (MAP) estimate of the parameters of the relative deformation between them. Our estimation process accomplishes two things simultaneously: It normalizes for pattern warping and it returns a distortion-tolerant similarity metric which can be used for matching two nonlinearly deformed image patterns. The prior probability of the deformation parameters is specific to the pattern-type and, therefore, should result in more accurate matching than an arbitrary general distribution. We show that the proposed method is very well suited for handling iris biometrics, applying it to two databases of iris images which contain real instances of warped patterns. We demonstrate a significant improvement in matching accuracy using the proposed deformed Bayesian matching methodology. We also show that the additional computation required to estimate the deformation is relatively inexpensive, making it suitable for real-time applications.


Rapid detection of methicillin resistance in coagulase-negative Staphylococci with the VITEK 2 system.
The aim of the present study was to evaluate the accuracy of the new VITEK 2 system (bioMérieux, Marcy l' Etoile, France) for the detection of methicillin resistance in coagulase-negative staphylococci (CoNS) by using AST-P515 and AST-P523 test cards. Analyses of the VITEK 2 oxacillin MIC determination evaluated according to the actual breakpoint (>/=0.5 micro g/ml) of the National Committee for Clinical Laboratory Standards resulted in a high sensitivity of 99.2% but a moderate specificity of 80%. The newly included oxacillin resistance (OR) test of the VITEK 2 system displayed a high sensitivity and a high specificity of 97.5 and 98.7%, respectively. Concordance between the results of the mecA PCR and the VITEK 2 oxacillin MIC was observed for almost all Staphylococcus epidermidis strains, but the reduced specificity was attributable to higher oxacillin MICs for mecA-negative non-S. epidermidis strains, especially S. saprophyticus, S. lugdunensis, and S. cohnii. Evaluation of alternative oxacillin MIC breakpoints of 1, 2, or 4 micro g/ml resulted in improved degrees of specificity of 84, 90.7, and 97.3%, respectively. Only minor changes occurred in the corresponding sensitivity values, which were 98.4, 97.5, and 97.5%, respectively. Methicillin resistance in CoNS was detected after 7 and 8 h in 91.1 and 93.5% of the mecA-positive strains, respectively, by the VITEK 2 OR test and in 86.3 and 89.5% of the mecA-positive strains, respectively, by VITEK 2 oxacillin MIC determination. After 7 and 8 h the VITEK 2 OR test classified 59.2 and 78.9% of the mecA-negative strains, respectively, as susceptible to oxacillin, whereas comparable values were obtained 2 h later by VITEK 2 oxacillin MIC determination. The results of our study encourage the use of the VITEK 2 system, which proved to be a highly reliable and rapid phenotypic method for the detection of methicillin resistance in CoNS.


Insulin pump use compared with intravenous insulin during labour and delivery: the INSPIRED observational cohort study.
To assess the safety and efficacy of pump therapy (continuous subcutaneous insulin infusion; CSII) during labour and delivery in women with Type 1 diabetes. A retrospective cohort study of 161 consecutive Type 1 diabetic pregnancies delivered during 2000-2010 at Mount Sinai Hospital, Toronto, Canada. Capillary blood glucose levels during labour and delivery and time in/out of target (target: 4-6 mmol/l) were compared along with neonatal outcomes for three groups: (1) women on pumps who stayed on pumps during labour (pump/pump n = 31), (2) women on pumps who switched to intravenous (IV) insulin infusion during labour (pump/IVn = 25), and (3) women on multiple daily injections who switched to IV insulin infusion during labour (MDIn = 105). There were no significant differences between the mean or median glucose values during labour and delivery across all three groups, and no significant difference in time spent hypoglycaemic. However, women in the pump/pump group had significantly better glycaemic control as defined by mean glucose (5.5 vs. 6.4 mmol/l; P = 0.01), median glucose (5.4 vs. 6.3 mmol/l; P = 0.02), and more time spent in target (60.9% vs. 39.2%; P = 0.06) compared with women in the pump/IV group (after removing one outlier). This study demonstrates that the continuation of CSII therapy during labour and delivery appears safe and efficacious. Moreover, women who choose to continue CSII have better glucose control during delivery than those who switch to IV insulin, suggesting that it should be standard practice to allow women the option of continuing CSII during labour and delivery.


The human small intestinal microbiota is driven by rapid uptake and conversion of simple carbohydrates.
The human gastrointestinal tract (GI tract) harbors a complex community of microbes. The microbiota composition varies between different locations in the GI tract, but most studies focus on the fecal microbiota, and that inhabiting the colonic mucosa. Consequently, little is known about the microbiota at other parts of the GI tract, which is especially true for the small intestine because of its limited accessibility. Here we deduce an ecological model of the microbiota composition and function in the small intestine, using complementing culture-independent approaches. Phylogenetic microarray analyses demonstrated that microbiota compositions that are typically found in effluent samples from ileostomists (subjects without a colon) can also be encountered in the small intestine of healthy individuals. Phylogenetic mapping of small intestinal metagenome of three different ileostomy effluent samples from a single individual indicated that Streptococcus sp., Escherichia coli, Clostridium sp. and high G+C organisms are most abundant in the small intestine. The compositions of these populations fluctuated in time and correlated to the short-chain fatty acids profiles that were determined in parallel. Comparative functional analysis with fecal metagenomes identified functions that are overrepresented in the small intestine, including simple carbohydrate transport phosphotransferase systems (PTS), central metabolism and biotin production. Moreover, metatranscriptome analysis supported high level in-situ expression of PTS and carbohydrate metabolic genes, especially those belonging to Streptococcus sp. Overall, our findings suggest that rapid uptake and fermentation of available carbohydrates contribute to maintaining the microbiota in the human small intestine.


The protofilament structure of insulin amyloid fibrils.
Under solution conditions where the native state is destabilized, the largely helical polypeptide hormone insulin readily aggregates to form amyloid fibrils with a characteristic cross-beta structure. However, there is a lack of information relating the 4.8 A beta-strand repeat to the higher order assembly of amyloid fibrils. We have used cryo-electron microscopy (EM), combining single particle analysis and helical reconstruction, to characterize these fibrils and to study the three-dimensional (3D) arrangement of their component protofilaments. Low-resolution 3D structures of fibrils containing 2, 4, and 6 protofilaments reveal a characteristic, compact shape of the insulin protofilament. Considerations of protofilament packing indicate that the cross-beta ribbon is composed of relatively flat beta-sheets rather than being the highly twisted, beta-coil structure previously suggested by analysis of globular protein folds. Comparison of the various fibril structures suggests that very small, local changes in beta-sheet twist are important in establishing the long-range coiling of the protofilaments into fibrils of diverse morphology.


Posterior cruciate ligament substitution is not essential for excellent postoperative outcomes in total knee arthroplasty.
The study compared postoperative range of motion (ROM) and functional outcome of total knee arthroplasty (TKA) with a posterior cruciate ligament (PCL)-substituting knee prosthesis compared with an ultracongruent PCL-sacrificing design. Two hundred nine patients underwent primary TKA. Posterior stabilized design (121) and highly conforming, PCL-sacrificing, ultracongruent design (88) TKAs were reviewed. Surgeon, surgical approach, and a hyperflexion postoperative rehabilitation protocol were the same. Results showed significant improvement in knee flexion, ROM, Knee Score, and Function Score within each group. Postoperative mean total ROM was slightly higher with the posterior stabilized design. The 2 groups were the same postoperatively in the improvement in ROM, Knee Score, Function Score, satisfaction level, among other activity metrics. There was no clear evidence proving superiority and the need for posterior stabilization in PCL-sacrificing TKA.


Are high generalised and asthma-specific self-efficacy predictive of adequate self-management behaviour among adult asthma patients?
In asthma self-management training, often self-treatment guidelines are included, because increased knowledge of asthma alone is not sufficient to change behaviour. One way to achieve behavioural changes is by increasing the patient's general and asthma-specific self-efficacy expectancies. This refers to beliefs in one's capabilities to execute the recommended course of action successfully. We wanted to assess whether high generalised and asthma-specific self-efficacy expectancies were predictive of adequate self-management and self-treatment behaviour. A questionnaire was sent to 4563 persons (18-65 years) who had used inhaled medication in 1993. Self-management and self-treatment behaviour were operationalised through a hypothetical scenario of a slow-onset asthma exacerbation. Of all 1262 asthmatic patients, 39.3% showed adequate self-treatment behaviour (self-adjusting their inhaled or oral steroids when appropriate). Age, asthma-specific outcome expectancies and knowledge were predictive of adequate self-treatment. Adequate self-management behaviour (self-treatment or seeking medical help) was observed in 56.4% of patients. Intentions towards self-management and asthma-specific knowledge were significant. Only knowledge has a relevant influence on both. Asthma-specific knowledge is the only factor that seems relevant for adequate self-management and self-treatment behaviour, which might be explained by the hypothetical nature of the scenario. When patients experience a real asthma exacerbation, self-efficacy expectancies will become more important. Only if knowledge of what to do is present will patients be able to show proper self-management and self-treatment behaviour. Our results suggest that self-treatment guidelines are only effective in combination with patient education, which is important for optimal control of their disease.


Multiphoton laser scanning microscopy for four-dimensional analysis of Caenorhabditis elegans embryonic development.
Multiphoton laser scanning microscopy (MPLSM) enables the production of long timelapse recordings from live fluorescent specimens. 1047- and 900-nm excitation were used to image both a vital fluorescent membrane probe, FM 4-64, and a modified green fluorescent protein (GFP) in live Caenorhabditis elegans embryos. Automated four-dimensional (4D) data collection yielded individual recordings comprising thousands of images, each allowing analysis of all of the cell divisions, contacts, migrations, and fusions that occur during a span of several hours of embryogenesis.


Preclinical evaluation of the PI3K/Akt/mTOR pathway in animal models of multiple sclerosis.
The PI3K/AKT/mTOR pathway is an intracellular signalling pathway that regulates cell activation. proliferation, metabolism and apoptosis. Increasing body of data suggests that alterations in the PI3K/AKT/mTOR pathway may result in an enhanced susceptibility to autoimmunity. Multiple Sclerosis (MS) is one of the most common chronic inflammatory diseases of the central nervous system leading to demyelination and neurodegeneration. In the current study, we have firstly evaluated in silico the involvement of the mTOR network on the generation and progression of MS and on oligodendrocyte function, making use of currently available whole-genome transcriptomic data. Then, the data generated in silico were subjected to an ex-vivo evaluation. To this aim, the involvement of mTOR was validated on a well-known animal model of MS and in vitro on Th17 cells. Our data indicate that there is a significant involvement of the mTOR network in the etiopathogenesis of MS and that Rapamycin treatment may represent a useful therapeutic approach in this clinical setting. On the other hand, our data showed that a significant involvement of the mTOR network could be observed only in the early phases of oligodendrocyte maturation, but not in the maturation process of adult oligodendrocytes and in the process of remyelination following demyelinating injury. Overall, our study suggests that targeting the PI3K/mTOR pathway, although it may not be a useful therapeutic approach to promote remyelination in MS patients, it can be exploited to exert immunomodulation, preventing/delaying relapses, and to treat MS patients in order to slow down the progression of disability.


TrkB inhibition as a therapeutic target for CNS-related disorders.
The interaction of brain-derived neurotrophic factor (BDNF) with its tropomyosin-related kinase receptor B (TrkB) is involved in fundamental cellular processes including neuronal proliferation, differentiation and survival as well as neurotransmitter release and synaptic plasticity. TrkB signaling has been widely associated with beneficial, trophic effects and many commonly used psychotropic drugs aim to increase BDNF levels in the brain. However, it is likely that a prolonged increased TrkB activation is observed in many pathological conditions, which may underlie the development and course of clinical symptoms. Interestingly, genetic and pharmacological studies aiming at decreasing TrkB activation in rodent models mimicking human pathology have demonstrated a promising therapeutic landscape for TrkB inhibitors in the treatment of various diseases, e.g. central nervous system (CNS) disorders and several types of cancer. Up to date, only a few selective and potent TrkB inhibitors have been developed. As such, the use of crystallography and in silico approaches to model BDNF-TrkB interaction and to generate relevant pharmacophores represent powerful tools to develop novel compounds targeting the TrkB receptor.


Living through conflict and post-conflict: experiences of health workers in northern Uganda and lessons for people-centred health systems.
Providing people-centred health systems--or any systems at all--requires specific measures to protect and retain healthcare workers during and after the conflict. This is particularly important when health staff are themselves the target of violence and abduction, as is often the case. This article presents the perspective of health workers who lived through conflict in four districts of northern Uganda--Pader, Gulu, Amuru, and Kitgum. These contained more than 90% of the people displaced by the decades of conflict, which ended in 2006. The article is based on 26 in-depth interviews, using a life history approach. This participatory tool encouraged participants to record key events and decisions in their lives, and to explore areas such as their decision to become a health worker, their employment history, and their experiences of conflict and coping strategies. These were analyzed thematically to develop an understanding of how to protect and retain staff in these challenging contexts. During the conflict, many health workers lost their lives or witnessed the death of their friends and colleagues. They also experienced abduction, ambush and injury. Other challenges included disconnection from social and professional support systems, displacement, limited supplies and equipment, increased workload and long working days and lack of pay. Health workers were not passive in the face of these challenges, however. They adopted a range of safety measures, such as mingling with community members, sleeping in the bush, and frequent change of sleeping place, in addition to psychological and practical coping strategies. Understanding their motivation and their views provides an important insight how to maintain staffing and so to continue to offer essential health care during difficult times and in marginalized areas.


25-Hydroxyvitamin D3 is a natural chemopreventive agent against carcinogen induced precancerous lesions in mouse mammary gland organ culture.
Despite the role of vitamin D(3) endocrine system in prevention of mammary gland transformation in animal models, use of 1,25(OH)(2)D(3 )in clinical settings is precluded due to its toxicity in vivo. Therefore much effort has been placed in developing relatively non-toxic vitamin D analogs. Recently, with the discovery of the expression of 25-hydroxy vitamin D(3) 1alpha-hydroxylase (CYP27B1) in multiple extrarenal organs, the functional role of prohormone, 25-hydroxyvitamin D(3) [25(OH)D(3)], has been redefined. Since 25(OH)D(3) does not cause hypercalcemia and maintains relative high concentration in serum, it is possible that the prohormone can be converted to active hormone in mammary epithelial cells to provide chemopreventive effects. In the present study, we evaluated its functional significance using mouse mammary organ culture (MMOC) system. We first showed that 25(OH)D(3) 1alpha-hydroxylase is extensively expressed in mammary ductal epithelial cells at both protein and mRNA levels, which is a prerequisite for 25(OH)D(3) to function in an autocrine/paracrine manner. However, we also observed that clotrimazol (1alpha-hydroxylase inhibitor) enhanced 25(OH)D(3) -induced CYP24 expression in breast cancer cells. In mammary glands derived from 1alpha-hydroxylase knockout mice, 25(OH)D(3) treatment in organ culture significantly induced CYP24 expression, indicating a potential direct effect of 25(OH)D(3). In MMOC, 100-250 nM 25(OH)D(3) suppressed both ovarian hormone-dependent and -independent mammary precancerous lesions (induced by DMBA) by more than 50%, while the active hormone 1,25(OH)(2)D(3) (positive control) at 100 nM suppressed alveolar lesions by more than 80%. The inactive vitamin D(3) (negative control) at 100 nM suppressed alveolar lesions by only 20% (P>0.05). We found that 25(OH)D(3) inhibits DMBA-induced mammary alveolar lesions (MAL) in a stage-specific manner: 25(OH)D(3) mainly inhibits the promotion stage of lesion formation. We conclude that 25(OH)D(3) could serve as a non-toxic natural chemopreventive agent for further development for breast cancer prevention.


Failure of postsynaptic specialization to develop at neuromuscular junctions of rapsyn-deficient mice.
Of numerous synaptic components that have been identified, perhaps the best-studied are the nicotinic acetylcholine receptors (AChRs) of the vertebrate neuromuscular junction. AChRs are diffusely distributed on embryonic myotubes, but become highly concentrated (approximately 10,000 microns-2) in the postsynaptic membrane as development proceeds. At least two distinct processes contribute to this accumulation. One is local synthesis: subsynaptic muscle nuclei transcribe AChR subunit genes at higher rates than extra-synaptic nuclei, so AChR messenger RNA is concentrated near synaptic sites. Second, once AChRs have been inserted in the membrane, they form high-density clusters by tethering to a subsynaptic cytoskeletal complex. A key component of this complex is rapsyn, a peripheral membrane protein of relative molecular mass 43K (refs 4, 5), which is precisely colocalized with AChRs at synaptic sites from the earliest stages of neuromuscular synaptogenesis. In heterologous systems, expression of recombinant rapsyn leads to clustering of diffusely distributed AChRs, suggesting that rapsyn may control formation of clusters. To assess the role of rapsyn in vivo, we generated and characterized mutant mice with a targeted disruption of the Rapsyn gene. We report that rapsyn is essential for the formation of AChR clusters, but that synapse-specific transcription of AChR subunit genes can proceed in its absence.