Subcortical Brain Alterations in Carriers of Genomic Copy Number Variants.
Copy number variants (CNVs) are well-known genetic pleiotropic risk factors for multiple neurodevelopmental and psychiatric disorders (NPDs), including autism (ASD) and schizophrenia. Little is known about how different CNVs conferring risk for the same condition may affect subcortical brain structures and how these alterations relate to the level of disease risk conferred by CNVs. To fill this gap, the authors investigated gross volume, vertex-level thickness, and surface maps of subcortical structures in 11 CNVs and six NPDs. Subcortical structures were characterized using harmonized ENIGMA protocols in 675 CNV carriers (CNVs at 1q21.1, TAR, 13q12.12, 15q11.2, 16p11.2, 16p13.11, and 22q11.2; age range, 6-80 years; 340 males) and 782 control subjects (age range, 6-80 years; 387 males) as well as ENIGMA summary statistics for ASD, schizophrenia, attention deficit hyperactivity disorder, obsessive-compulsive disorder, bipolar disorder, and major depression. All CNVs showed alterations in at least one subcortical measure. Each structure was affected by at least two CNVs, and the hippocampus and amygdala were affected by five. Shape analyses detected subregional alterations that were averaged out in volume analyses. A common latent dimension was identified, characterized by opposing effects on the hippocampus/amygdala and putamen/pallidum, across CNVs and across NPDs. Effect sizes of CNVs on subcortical volume, thickness, and local surface area were correlated with their previously reported effect sizes on cognition and risk for ASD and schizophrenia. The findings demonstrate that subcortical alterations associated with CNVs show varying levels of similarities with those associated with neuropsychiatric conditions, as well distinct effects, with some CNVs clustering with adult-onset conditions and others with ASD. These findings provide insight into the long-standing questions of why CNVs at different genomic loci increase the risk for the same NPD and why a single CNV increases the risk for a diverse set of NPDs.


Determinants and prognostic significance of an intra-dialysis rise of cardiac troponin I measured by sensitive assay in hemodialysis patients.
Previous studies using conventional cTnI assays reported conflicting results on the evolution of cTnI levels during hemodialysis. The determinants and prognostic significance of changes in cTnI during hemodialysis are presently unknown. The aim of this prospective study was to characterize the determinants and prognostic significance of intra-dialysis changes in cTnI using a sensitive assay. cTnI was measured before and after hemodialysis with a sensitive assay in 90 chronic patients without acute cardiac symptoms. Multivariable regression analyses were used to identify factors that were associated with intra-dialysis rise in cTnI. The prognostic effect of an intra-dialysis rise in cTnI during a 52-month follow-up was evaluated using Cox regression models. The primary and secondary endpoint was the incidence of major adverse cardiovascular events and all-cause mortality, respectively. Pre-dialysis cTnI was elevated in 31 patients (34 %). cTnI increased significantly during dialysis and this had a trend to be associated with longer dialysis vintage. A greater intra-dialysis rise in cTnI was associated with a significantly higher incidence of cardiovascular events, also after correction for age, gender, dialysis vintage, residual diuresis, previous cardiovascular events, and pre-dialysis cTnI levels (HR per 10 ng/L rise in cTnI: 1.21; CI 1.06-1.38; p 0.005). TnI levels rise significantly during hemodialysis and a greater intra-dialysis rise in cTnI is associated with an increased incidence of cardiovascular events. These findings suggest that hemodialysis has an acute deleterious effect on the heart. An intra-dialysis rise in cTnI may help identify patients who are susceptible to the hemodynamic stress of hemodialysis.


Parents' and teachers' ratings of problem behaviours in children: genetic and contrast effects.
We obtained ratings on the Conners' scales from teachers (CTRS-28) and parents (CPRS-48) for 61 monozygotic and 64 dizygotic twin pairs, aged between 7 and 11 years. Model-fitting analyses were carried out to estimate the extent of genetic and environmental influences on problem behaviours, and to explore possible contrast effects in ratings by parents and teachers. Confirming previous findings with other measures, there was evidence of moderate to strong genetic effects on a range of problem behaviours. Parents' ratings on the Anxiety, Impulsive-Hyperactive and Learning Problem sub-scales showed significant evidence of contrast effects. There was no evidence of such rater bias or competitive sibling interaction effects in ratings by teachers, or in parents' ratings on the Conduct Problem and Psychosomatic sub-scales.


Design and Implementation of a Low-Cost Priapism Reduction Task Trainer.
This low-cost priapism reduction task trainer is designed to instruct emergency medicine (EM) resident physicians. Priapism is a true urologic emergency that EM physicians must be able to diagnose and treat in order to prevent significant tissue damage and loss of erectile function. Given the nature of the condition, priapism treatment is often both physically invasive and psychologically upsetting. Simulation allows learners to practice invasive or rare procedures in a safe and educational environment. At present, there are few inexpensive and easily created task trainers for priapism reduction. Our goal was to create an inexpensive, reusable task trainer that allows learners to practice the skills needed for priapism reduction. By the end of this educational session, learners should be able to 1) Verbalize the difference between low-flow and high-flow priapism 2) Describe the landmarks for a penile ring block and cavernosal aspiration/injection 3) Demonstrate the appropriate technique for performing a penile ring block, cavernosal aspiration, and cavernosal injection. Using inexpensive and commonly found materials, we were able to successfully create a partial task trainer for teaching priapism reduction techniques including administering local anesthesia, medication injection, and realistic cavernosal aspiration with simulated blood return. As part of a standard EM residency didactics curriculum, this task trainer has been used to teach post graduate year (PGY) 1-4 resident learners. After an introductory didactic session, participants were given the opportunity for hands-on skills-based practice using the simulated task trainer. Learners were asked to complete a post-session survey to assess the educational value of the station and the task trainer. We were able to successfully create a low cost, easy to build partial task trainer for priapism reduction that allowed learners to perform local anesthesia, medication injections, and corporal aspiration. Twenty-five residents (ten PGY-1, five PGY-2, five PGY-3, five PGY-4) participated in a single didactic session and completed a post-session survey. The majority (68%, N=17) of participants had never previously treated a patient with priapism. On average, participants rated their comfort managing a patient with priapism before the session to be 1.76 on a 5-point Likert-scale (where 1=not at all comfortable and 5=extremely comfortable). Following the session, participants' comfort increased to 3.76 on the same scale. Participants rated the usefulness of the priapism model for teaching priapism reduction techniques to be 4.64 on a 5- point scale (where 1=not at all useful and 5=extremely useful). Using inexpensive and commonly found materials, we were able to successfully create a partial task trainer for teaching priapism reduction techniques including local anesthesia, medication injection, and cavernosal aspiration. Learners reported that the educational session greatly increased their confidence in caring for patients presenting with priapism. Additionally, they found the priapism model to be extremely helpful for teaching reduction techniques. Our model was successful in teaching a procedure that providers may encounter in clinical practice yet most resident learners had not yet had the opportunity to perform in training. Penile anesthesia, priapism reduction, urologic emergencies.


Effect of treatment of a previous ectopic pregnancy on in vitro fertilization-intracytoplasmic sperm injection outcomes: a retrospective cohort study.
To evaluate the influence of previous ectopic pregnancies (EP) and different EP treatment methods on in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI) outcomes. Retrospective cohort study. University-based reproductive medicine center. Women undergoing 51,268 fresh cycles of IVF-ICSI. In women who had had an EP before the IVF treatment and women who were diagnosed with fallopian disease without an EP (control group), division into three treatment subgroups: subgroup 1, conservative treatment; subgroup 2, salpingostomy; and subgroup 3 salpingectomy. Estimates of IVF-ICSI outcome and ovarian function based on number of antral follicles and rates of implantation, EP, clinical pregnancy, and delivery. For the experimental group and the control group, there were no statistically significant differences in the rates of implantation, clinical pregnancy, or delivery. However, the risk of recurrent EP was statistically significantly higher in the experimental group compared with the control group. There were no statistically significant differences in IVF-ICSI outcomes between the three treatment subgroups in terms of the basic antral follicle number or the dominant antral follicle numbers (>1.0 cm) after surgical treatment of EP. Our results suggest that women with a prior history of EP have a higher recurrence risk of EP after IVF in comparison with women with no history of EP. Previous EP has no effect on the main IVF-ICSI outcomes. For the treatment of EP, salpingostomy and salpingectomy do not statistically significantly affect ovarian function.


Reductive dehydrocoupling of diphenyltin dihydride with LiAlH4: selective synthesis and structures of the first bicyclo[2.2.1]heptastannane-1,4-diide and bicyclo[2.2.2]octastannane-1,4-diide.
The reaction of diphenyltin dihydride with LiAlH4 gives access to a set of charged tin cages as their lithium salts. Variation in the ratio of reactants provides a perstannabicyclooctane dianion and a perstannanorbornane as the di- and monoanions. These compounds can be synthesised selectively by careful stoichiometric control and have been characterised by single crystal X-ray diffractometry, NMR and UV-vis spectroscopy. Computational exploration of the electronic structures of these compounds was undertaken and, in agreement with structural and spectroscopic features, indicated significant σ-delocalisation in the tin skeletons.


Mosquito fauna and breeding habitats of anophelines in Little Andaman Island, Andaman and Nicobar Islands, India.
A total of 37 species of mosquitoes belonging to nine genera--Anopheles, Aedes, Armigeres, Culex, Harpagomyia, Mansonia, Orthopodomyia, Toxorhynchites and Uranotaenia were collected from Little Andaman Island. Mosquitoes of nine Anopheles group of species--An. barbirostris, An. barbumbrosus, An. balabacensis, An. insulaeflorum, An. kochi, An. philippinensis, An. roperi, An. sundaicus and An. vagus were recorded. An. philippinensis was found to be the most predominant species. Species-specific breeding preference and association in various types of acquatic habitats have been reported. This is the first record of mosquito fauna of Little Andaman Island.


Activation of Tm-22 resistance is mediated by a conserved cysteine essential for tobacco mosaic virus movement.
The tomato Tm-22 gene was considered to be one of the most durable resistance genes in agriculture, protecting against viruses of the Tobamovirus genus, such as tomato mosaic virus (ToMV) and tobacco mosaic virus (TMV). However, an emerging tobamovirus, tomato brown rugose fruit virus (ToBRFV), has overcome Tm-22 , damaging tomato production worldwide. Tm-22 encodes a nucleotide-binding leucine-rich repeat (NLR) class immune receptor that recognizes its effector, the tobamovirus movement protein (MP). Previously, we found that ToBRFV MP (MPToBRFV ) enabled the virus to overcome Tm-22 -mediated resistance. Yet, it was unknown how Tm-22 remained durable against other tobamoviruses, such as TMV and ToMV, for over 60 years. Here, we show that a conserved cysteine (C68) in the MP of TMV (MPTMV ) plays a dual role in Tm-22 activation and viral movement. Substitution of MPToBRFV amino acid H67 with the corresponding amino acid in MPTMV (C68) activated Tm-22 -mediated resistance. However, replacement of C68 in TMV and ToMV disabled the infectivity of both viruses. Phylogenetic and structural prediction analysis revealed that C68 is conserved among all Solanaceae-infecting tobamoviruses except ToBRFV and localizes to a predicted jelly-roll fold common to various MPs. Cell-to-cell and subcellular movement analysis showed that C68 is required for the movement of TMV by regulating the MP interaction with the endoplasmic reticulum and targeting it to plasmodesmata. The dual role of C68 in viral movement and Tm-22 immune activation could explain how TMV was unable to overcome this resistance for such a long period.


An overview of regular dialysis treatment in Japan (as of 31 December 2004).
A statistical survey of 3932 nationwide hemodialysis (hereafter, dialysis) facilities was carried out at the end of 2004, and 3882 facilities (98.73%) responded. The population undergoing dialysis at the end of 2004 was 248 166, an increase of 10 456 patients (4.4%) from that at the end of 2003. The number of dialysis patients per million people was 1943.5. The crude death rate of dialysis patients from the end of 2003 to the end of 2004 was 9.4%. The mean age of patients who underwent dialysis in 2004 was 65.8 years, and that of the total dialysis population was 63.3 years. The percentage distribution of patients who underwent dialysis according to a newly underlying disease showed that 41.3% of patients had diabetic nephropathy and 28.1% had chronic glomerulonephritis. The frequency of calcium carbonate use for dialysis patients was 75.1% and that of sevelamer hydrochloride use was 26.2%. The frequency of sevelamer hydrochloride use does not necessarily have a strong correlation with the dose of calcium carbonate. Patients who received high doses of sevelamer hydrochloride tended to have a low concentration of arterial blood HCO(3-). Approximately 15% of dialysis patients used an intravenous vitamin D preparation, generally maxacalcitol. The longer the patients had been on dialysis, the higher the frequency of use of an intravenous vitamin D preparation. When the concentration of serum intact parathyroid hormone (PTH) was more than 200 pg/mL, the frequency of use of an orally administered vitamin D preparation decreased; but that of intravenous vitamin D preparation increased. The percentage of dialysis patients who received percutaneous ethanol injection therapy (PEIT) was 1.4%. The percentage was more than 50% in the patients who had been on dialysis for more than 10 years. The percentage of patients who received PEIT again was 35.0%. The percentage of patients who had been on hemodialysis for more than 10 years and received PEIT again was more than 50%.


Bilateral peripheral neural activity observed in vivo following unilateral nerve injury.
Manganese-enhanced magnetic resonance imaging (MRI) is a surrogate method to measure calcium content in nervous system since manganese physiologically follows calcium. Manganese is detectable in MRI and therefore visualizes structures and cell populations that actively regulate calcium. Since calcium is actively recruited for the transmission of action potentials, our purpose is to validate manganese-enhanced MRI for detection of changes in lumbar nerves related to nociception. A neuropathic pain model was created by chronic constrictive injury of the left sciatic nerve of Sprague-Dawley rats. Behavioral measurements, using von Frey's tests, confirmed the presence of significant allodynia in the left hind limb of animals in the injured group. T1-weighted fast spin echo images were obtained of the lumbar cord and plexus of animals with injured left sciatic nerve and uninjured animals (control) scanned in a 7 Tesla magnet after intraperitoneal manganese chloride administration four weeks after surgery. Lumbar nerve roots and sciatic nerves in the injured group show increased normalized manganese-enhanced MRI signal, representing manganese enhancement, compared to the control group. In conclusion, animals with neuropathic pain in the left hind limb show increased manganese uptake in not only the injured sciatic nerve but also in the contralateral uninjured sciatic nerve on manganese-enhanced MRI in vivo. Although poorly understood, this finding corroborates ex vivo finding of bilateral nociceptive-related molecular changes in the nervous system of unilateral pain models.


Prolonged Adaptive Evolution of a Defensive Gene in the Solanaceae.
Although plants and their natural enemies may coevolve for prolonged periods, little is known about how long individual plant defensive genes are involved in the coevolutionary process. We address this issue by examining patterns of selection on the defensive gene threonine deaminase (TD). Tomato (Solanum lycopersicum) has two copies of this gene. One performs the canonical housekeeping function in amino acid metabolism of catalyzing the first reaction in the conversion of threonine to isoleucine. The second copy functions as an antinutritive defense against lepidopteran herbivores by depleting threonine in the insect gut. Wild tobacco (Nicotiana attenuata) also contains a defensive copy. We show that a single copy of TD underwent two or three duplications near the base of the Solanaceae. One copy retains the housekeeping function, whereas a second copy evolved defensive functions. Positive selection occurred on the branch of the TD2 gene tree subtending the common ancestor of the Nicotianoideae and Solanoideae. It also occurred within the Solanoideae clade but not within the Nicotianoideae clade. Finally, it occurred on most branches leading from the common ancestor to S. lycopersicum. Based on recent calibrations of the Solanaceae phylogeny, TD2 experienced adaptive substitutions for a period of 30-50 My. We suggest that the most likely explanation for this result is fluctuating herbivore abundances: When herbivores are rare, relaxed selection increases the likelihood that slightly disadvantageous mutations will be fixed by drift; when herbivores are common, increased selection causes the evolution of compensatory adaptive mutations. Alternative explanations are also discussed.


A Comparison of South African National HIV Incidence Estimates: A Critical Appraisal of Different Methods.
The interpretation of HIV prevalence trends is increasingly difficult as antiretroviral treatment programs expand. Reliable HIV incidence estimates are critical to monitoring transmission trends and guiding an effective national response to the epidemic. We used a range of methods to estimate HIV incidence in South Africa: (i) an incidence testing algorithm applying the Limiting-Antigen Avidity Assay (LAg-Avidity EIA) in combination with antiretroviral drug and HIV viral load testing; (ii) a modelling technique based on the synthetic cohort principle; and (iii) two dynamic mathematical models, the EPP/Spectrum model package and the Thembisa model. Overall, the different incidence estimation methods were in broad agreement on HIV incidence estimates among persons aged 15-49 years in 2012. The assay-based method produced slightly higher estimates of incidence, 1.72% (95% CI 1.38 - 2.06), compared with the mathematical models, 1.47% (95% CI 1.23 - 1.72) in Thembisa and 1.52% (95% CI 1.43 - 1.62) in EPP/Spectrum, and slightly lower estimates of incidence compared to the synthetic cohort, 1.9% (95% CI 0.8 - 3.1) over the period from 2008 to 2012. Among youth aged 15-24 years, a declining trend in HIV incidence was estimated by all three mathematical estimation methods. The multi-method comparison showed similar levels and trends in HIV incidence and validated the estimates provided by the assay-based incidence testing algorithm. Our results confirm that South Africa is the country with the largest number of new HIV infections in the world, with about 1 000 new infections occurring each day among adults aged 15-49 years in 2012.


Shared and Distinct Features of Human Milk and Infant Stool Viromes.
Infants acquire many of their microbes from their mothers during the birth process. The acquisition of these microbes is believed to be critical in the development of the infant immune system. Bacteria also are transmitted to the infant through breastfeeding, and help to form the microbiome of the infant gastrointestinal (GI) tract; it is unknown whether viruses in human milk serve to establish an infant GI virome. We examined the virome contents of milk and infant stool in a cohort of mother-infant pairs to discern whether milk viruses colonize the infant GI tract. We observed greater viral alpha diversity in milk than in infant stool, similar to the trend we found for bacterial communities from both sites. When comparing beta diversity, viral communities were mostly distinguishable between milk and infant stool, but each was quite distinct from adult stool, urine, and salivary viromes. There were significant differences in viral families in the infant stool (abundant bacteriophages from the family Siphoviridae) compared to milk (abundant bacteriophages from the family Myoviridae), which may reflect significant differences in the bacterial families identified from both sites. Despite the differences in viral taxonomy, we identified a significant number of shared viruses in the milk and stool from all mother-infant pairs. Because of the significant proportion of bacteriophages transmitted in these mother-infant pairs, we believe the transmission of milk phages to the infant GI tract may help to shape the infant GI microbiome.


Active nuclear import and passive nuclear export are the primary determinants of TDP-43 localization.
ALS (Amyotrophic Lateral Sclerosis) is a neurodegenerative disease characterized by the redistribution of the RNA binding protein TDP-43 in affected neurons: from predominantly nuclear to aggregated in the cytosol. However, the determinants of TDP-43 localization and the cellular insults that promote redistribution are incompletely understood. Here, we show that the putative Nuclear Export Signal (NES) is not required for nuclear egress of TDP-43. Moreover, when the TDP-43 domain which contains the putative NES is fused to a reporter protein, YFP, the presence of the NES is not sufficient to mediate nuclear exclusion of the fusion protein. We find that the previously studied "∆NES" mutant, in which conserved hydrophobic residues are mutated to alanines, disrupts both solubility and splicing function. We further show that nuclear export of TDP-43 is independent of the exportin XPO1. Finally, we provide evidence that nuclear egress of TDP-43 is size dependent; nuclear export of dTomato TDP-43 is significantly impaired compared to Flag TDP-43. Together, these results suggest nuclear export of TDP-43 is predominantly driven by passive diffusion.


Special issue: Catalytic mechanisms by biological systems--introduction.
Research on enzyme mechanisms is advancing knowledge of the chemistry and biochemistry of catalytic mechanisms by biological systems. The structural-dynamical properties of enzymes are of key importance. Advanced methodological approaches and new insights into enzyme functioning, and new emerging approaches for the redesign/design of enzymes or artificial biocatalysts, were discussed at the EMBO conference on Catalytic Mechanisms by Biological Systems.


Association between memory B-cells and clinical and immunological features of primary Sjögren's syndrome and Sicca patients.
B-cells play a pivotal role in primary Sjögren's syndrome (pSS) pathogenesis. We aim to (1) evaluate the distribution of B-lymphocyte subpopulations in pSS and Sicca patients, (2) establish cut-off points that discriminate pSS from controls, (3) evaluate the association between memory B-cells and phenotypic features in pSS. We included 57 pSS patients, 68 Sicca and 24 healthy controls. Circulating B-cells were characterized by flow cytometry as naïve and memory subsets and classified from Bm1 to Bm5. Compared to controls, pSS patients had lower percentages (29.5 vs 44.4%) and absolute numbers (47 vs 106 cells/µl) of memory B-cells. Through ROC curves, a cut-off of ≤ 58 total memory B-cells/µl yielded a specificity of 0.88 and a sensitivity of 0.60 for pSS, and was met by 59.6% of pSS patients, 38.8% of Sicca and 12.5% of controls. A cut-off of < 23.5 Switched-memory B-cells/µl yielded a specificity of 0.88 and a sensitivity of 0.54 and was met by 54.4% of pSS patients, 37.3% of Sicca and 12.5% of controls. In pSS, lower total memory B-cells count was associated with longer disease duration (14.3 vs 8.1 years, p = 0.006) and more active disease profile, as evaluated by the European League Against Rheumatism (EULAR) Sjögren's Syndrome Disease Activity Index (ESSDAI) (3.1 vs 1.4, p = 0.043). Decreased numbers of memory B-cells clearly discriminated pSS from controls and can also have prognostic value. It remains to be clarified whether Sicca patients with decreased memory B-cells represent pSS and if B-cell profiling could help in the diagnosis of pSS.


Reducing lower extremity amputations due to diabetes: the application of diabetic-foot protocol in Chiang Mai University Hospital.
The aim of this study was to determine whether intensive treatment and education strategies for diabetic patients with ulcers help in preventing leg amputation. From August 2005 to March 2007, a diabetic-foot protocol using a multidisciplinary approach was applied at our hospital. All the subjects were educated regarding diabetic-foot disease and its complications and prevention. This report compares the amputation rate in patients receiving the protocol care from August 2005 to March 2007 with those who had standard care from August 2003 to July 2005. Seventy-three and 110 diabetic-foot ulcer patients received protocol and standard foot care, respectively. The incidence of major amputations in the protocol and standard care groups was 4.1% and 13.6%, respectively (P= .03). Our protocol was associated with improved diabetic-foot care outcomes. It can be used by any hospital to improve outcomes for patients with diabetes.


Self-reported psychotic-like experiences are a poor estimate of clinician-rated attenuated and frank delusions and hallucinations.
One reason for the decision to delay the introduction of an Attenuated Psychosis Syndrome in the main text of the fifth edition of the American Psychiatric Association's Diagnostic and Statistical Manual of Mental Disorders was the concern that attenuated psychotic symptoms (APS) might in fact be common features in adolescents and young adults from the general population of no psychopathological significance in themselves. This concern was based on reports of high prevalence rates of psychotic-like experiences (PLEs) in the general population and the assumption that PLEs are a good estimate of APS. Although the criterion validity of self-reported PLEs had already been studied with respect to clinician-rated psychotic symptoms and found insufficient, it had been argued that PLEs might in fact be more comparable with mild, subclinical expressions of psychotic symptoms and, therefore, with APS. The present paper is the first to specifically study this assumption. The sample consisted of 123 persons seeking help at a service for the early detection of psychosis, of whom 54 had an at-risk mental state or psychosis, 55 had a nonpsychotic mental disorder and 14 had no full-blown mental disorder. PLEs were assessed with the Peters Delusion Inventory and the revised Launay-Slade Hallucination Scale, and psychotic symptoms and APS were assessed with the Structured Interview for Prodromal Syndromes. At a level of agreement between the presence of any PLE (in 98.4% of patients) and any APS (in 40.7%) just exceeding chance (κ = 0.022), the criterion validity of PLEs for APS was insufficient. Even if additional qualifiers (high agreement or distress, preoccupation and conviction) were considered, PLEs (in 52.8%) still tended to significantly overestimate APS, and agreement was only fair (κ = 0.340). Furthermore, the group effect on PLE prevalence was, at most, moderate (Cramer's V ≤ 0.382). The prevalence of APS cannot be deduced from studies of PLEs. Thus, the high population prevalence rate of PLEs does not allow the conclusion that APS are common features of no pathological significance and would lack clinical validity as an Attenuated Psychosis Syndrome in the Diagnostic and Statistical Manual of Mental Disorders, 5th edition. Rather, the population prevalence rate of APS has to be assumed to be largely unknown at present but is likely lower than indicated by epidemiological studies of PLEs. Therefore, dedicated studies are warranted, in which APS are assessed in a way that equates to their clinical evaluation.


Impact of macrophage inflammatory protein-1α deficiency on atherosclerotic lesion formation, hepatic steatosis, and adipose tissue expansion.
Macrophage inflammatory protein-1α (CCL3) plays a well-known role in infectious and viral diseases; however, its contribution to atherosclerotic lesion formation and lipid metabolism has not been determined. Low density lipoprotein receptor deficient (LDLR(-/-)) mice were transplanted with bone marrow from CCL3(-/-) or C57BL/6 wild type donors. After 6 and 12 weeks on western diet (WD), recipients of CCL3(-/-) marrow demonstrated lower plasma cholesterol and triglyceride concentrations compared to recipients of C57BL/6 marrow. Atherosclerotic lesion area was significantly lower in female CCL3(-/-) recipients after 6 weeks and in male CCL3(-/-) recipients after 12 weeks of WD feeding (P<0.05). Surprisingly, male CCL3(-/-) recipients had a 50% decrease in adipose tissue mass after WD-feeding, and plasma insulin, and leptin levels were also significantly lower. These results were specific to CCL3, as LDLR(-/-) recipients of monocyte chemoattractant protein(-/-) (CCL2) marrow were not protected from the metabolic consequences of high fat feeding. Despite these improvements in LDLR(-/-) recipients of CCL3(-/-) marrow in the bone marrow transplantation (BMT) model, double knockout mice, globally deficient in both proteins, did not have decreased body weight, plasma lipids, or atherosclerosis compared with LDLR(-/-) controls. Finally, there were no differences in myeloid progenitors or leukocyte populations, indicating that changes in body weight and plasma lipids in CCL3(-/-) recipients was not due to differences in hematopoiesis. Taken together, these data implicate a role for CCL3 in lipid metabolism in hyperlipidemic mice following hematopoietic reconstitution.


A comprehensive characterization of cell-free RNA in spent blastocyst medium and quality prediction for blastocyst.
The rate of pregnancy can be affected by many factors in assisted reproductive technology (ART), and one of which is the quality of embryos. Therefore, selecting the embryos with high potential is crucial for the outcome. Fifteen spent blastocyst medium (SBM) samples were collected from 14 patients who received in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), seven from high-grade embryos and eight from low-grade embryos. Cell-free RNA (cf-RNA) profile of SBM samples were analyzed by RNA sequencing in the present study. It was found that a large amount of cf-RNA were released into SBM, including protein-coding genes (68.9%) and long noncoding RNAs (lncRNAs) (17.26%). Furthermore, a high correlation was observed between blastocyst genes and SBM genes. And the cf-mRNAs of SBM were highly fragmented, and coding sequence (CDS) and untranslated (UTR) regions were released equally. Two hundred and thirty-two differentially expressed genes were identified in high-grade SBM (hSBM) and low-grade SBM (lSBM), which could be potential biomarker in distinguishing the embryos with different quality as an alternative or supplementary approach for subjective morphology criteria. Hence, cf-RNAs sequencing revealed the characterization of circulating transcriptomes of embryos with different quality. Based on the results, the genes related to blastocyst quality were screened, including the genes closely related to translation, immune-signaling pathway, and amino acid metabolism. Overall, the present study showed the types of SBM cf-RNAs, and the integrated analysis of cf-RNAs profiling with morphology grading displayed its potential in predicting blastocyst quality. The present study provided valuable scientific basis for noninvasive embryo selection in ART by RNA-profiling analysis.


Treatment of mycosis fungoides with total skin electron beam: response and relapse by ethnicity and sex.
To determine whether clinical response to total skin electron beam (TSEB) and relapse after TSEB differs by ethnicity and sex. Retrospective chart review of 77 patients with mycosis fungoides (MF), treated with TSEB in 2002 to 2008 at Yale University School of Medicine, Departments of Dermatology and Therapeutic Radiology. Women had better odds of response to TSEB than men (OR=6.4; 95% CI, 1.45-28.5; P=0.01). No significant difference was observed in response to TSEB between white and black patients (OR=0.69; 95% CI, 0.16-2.91; P=0.62). When stratified by race and sex, in comparison with black females, all other groups had lower odds of complete response (CR) to TSEB: black males (OR=0.39; 95% CI, 0.002-0.70; P=0.03), white females (OR=0.24; 95% CI, 0.02-2.53; P=0.24), and white males (OR=0.06; 95% CI, 0.006-0.60; P=0.02). Clinical CR was significantly predicted by the duration of symptoms (OR=0.98; 95% CI, 0.97-0.99; P=0.01); and nearly significant by clinical stage; stage III to stage I (OR=0.17; 95% CI, 0.02-1.02; P=0.07). Adjuvant treatment, previous treatment, and time from diagnosis to treatment have no significant effect on CR to TSEB. There was no statistically significant association between relapse after treatment and race, sex, clinical stage, or symptom duration. The odds of achieving a CR to TSEB decrease when diagnosis of MF is delayed and when patients present with advanced-stage disease. Women with MF were more likely to have a CR to treatment, and this response was even more significant in black women.


Reducing the Risk of Transmission of Critical Antimicrobial Resistance Determinants From Contaminated Pork Products to Humans in South-East Asia.
Antimicrobial resistance (AMR) is a critical challenge worldwide as it impacts public health, especially via contamination in the food chain and in healthcare-associated infections. In relation to farming, the systems used, waste management on farms, and the production line process are all determinants reflecting the risk of AMR emergence and rate of contamination of foodstuffs. This review focuses on South East Asia (SEA), which contains diverse regions covering 11 countries, each having different levels of development, customs, laws, and regulations. Routinely, here as elsewhere antimicrobials are still used for three indications: therapy, prevention, and growth promotion, and these are the fundamental drivers of AMR development and persistence. The accuracy of detection of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARG) depends on the laboratory standards applicable in the various institutes and countries, and this affects the consistency of regional data. Enterobacteriaceae such as Escherichia coli and Klebsiella pneumoniae are the standard proxy species used for indicating AMR-associated nosocomial infections and healthcare-associated infections. Pig feces and wastewater have been suspected as one of the hotspots for spread and circulation of ARB and ARG. As part of AMR surveillance in a One Health approach, clonal typing is used to identify bacterial clonal transmission from the production process to consumers and patients - although to date there have been few published definitive studies about this in SEA. Various alternatives to antibiotics are available to reduce antibiotic use on farms. Certain of these alternatives together with improved disease prevention methods are essential tools to reduce antimicrobial usage in swine farms and to support global policy. This review highlights evidence for potential transfer of resistant bacteria from food animals to humans, and awareness and understanding of AMR through a description of the occurrence of AMR in pig farm food chains under SEA management systems. The latter includes a description of standard pig farming practices, detection of AMR and clonal analysis of bacteria, and AMR in the food chain and associated environments. Finally, the possibility of using alternatives to antibiotics and improving policies for future strategies in combating AMR in a SEA context are outlined.


The purification of tissue inhibitor of metalloproteinases-2 from its 72 kDa progelatinase complex. Demonstration of the biochemical similarities of tissue inhibitor of metalloproteinases-2 and tissue inhibitor of metalloproteinases-1.
Human gingival fibroblasts in culture were shown to secrete a 72 kDa progelatinase, of which a proportion in the medium was found to be complexed with tissue inhibitor of metalloproteinases-2 (TIMP-2). A purification procedure was devised to purify free enzyme and inhibitor. We also describe the purification of both 95 kDa progelatinase bound to TIMP-1 and free 95 kDa progelatinase from the medium of U937 cells. A polyclonal antiserum to TIMP-2 was prepared and it was shown that TIMP-1 and TIMP-2 are antigenically distinct. The ability to form stable complexes and the relative inhibitory activities of TIMP-1 and TIMP-2 towards 95 kDa and 72 kDa gelatinases, collagenase, stromelysins 1 and 2 and punctuated metalloproteinase were determined; only minor differences were found. Complex-formation between TIMP-2 and 72 kDa progelatinase was demonstrated not to reduce the metalloproteinase-inhibitory activity of TIMP-2, a finding that led to the characterization of high-molecular-mass TIMP activity. Competition experiments between progelatinases and active gelatinases for TIMPs indicated that the affinity of TIMPs for progelatinases is weaker than that for active gelatinases. In a study of the effects of TIMP-1 and TIMP-2 on progelatinase self-cleavage we found that both TIMP-1 and TIMP-2 inhibit the conversion of 95 kDa and 72 kDa progelatinases and prostromelysin into lower-molecular-mass forms. TIMP capable of complexing with progelatinase was shown to be no more efficient an inhibitor of gelatinase self-cleavage than TIMP not able to complex with progelatinase.


Growth and body composition in preterm infants with bronchopulmonary dysplasia.
To compare growth and body composition in preterm infants with bronchopulmonary dysplasia (BPD) with normal healthy term infants during the first year of life. Twenty nine preterm infants with BPD (mean (SD) gestational age 27.1 (1.6) weeks; birth weight 852 (173) g) were followed prospectively. Anthropometry and body composition determined by total body electrical conductivity were measured and compared with those of healthy term infants at the same post-term age. In infants with BPD, the mean weight standard deviation scores (SD scores) 6 weeks after term were significantly lower (-1.44 and -2.68, boys and girls respectively) than in healthy term infants of the same age and did not improve during the first year. The mean length SD score was significantly lower in infants with BPD 6 weeks after term than in healthy term infants of the same age, and, although it improved significantly during the first year, the mean length SD score in girls with BPD was significantly below 0 12 months after term. In infants with BPD, the mean free fat mass (FFM) SD score and the mean total body fat (TBF) SD score at 6 weeks post-term age were significantly below 0. The mean FFM SD scores (-1.01 and -2.56, boys and girls respectively) and the mean TBF SD scores (-1.14 and -2.40, boys and girls respectively) 12 months after term were significantly lower than in healthy term infants of the same age. Preterm infants with BPD have impaired growth, with a deficit in TBF and FFM already 6 weeks after term; FFM and TBF remain low compared with healthy term infants during the first year of life. Nutritional intervention studies in infants with BPD are needed to evaluate if nutrition is the major determinant of growth and body composition or if this pattern of growth in preterm infants with BPD is the result of disturbed endocrine control.


Safety outcomes of anti-platelet therapy post coronary artery bypass graft surgery: A systematic review and network meta-analysis of randomized control trials.
Antiplatelet therapy is used to decrease the risk of graft failure post coronary artery bypass graft surgery. We aimed to compare dual antiplatelet therapy (DAPT) with monotherapy along with a comparison of Aspirin, Ticagrelor, Aspirin+Ticagrelor (A+T) and Aspirin+Clopidogrel (A+C) to determine the major and minor bleeding risk, risk of postoperative myocardial infarction (MI), stroke, and all-cause mortality (ACM). Randomized Controlled Trials comparing the four groups were included. Odds ratio (OR) and Absolute Risk (AR) were employed to assess the mean and standard deviation (SD) with 95% confidence intervals (CI). The Bayesian random-effects model was used for statistical analysis. Risk difference and Cochran Q tests were used to calculate rank probability (RP) and heterogeneity, respectively. We included 10 trials, consisting of 21 arms and 3926 patients. For the risk of major and minor bleed, A + T and Ticagrelor showed the lowest mean value of 0.040 (0.043) and 0.067 (0.073), respectively, and the highest RP of being the safest group. While a direct comparison between DAPT and monotherapy resulted in an OR of 0.57 [0.34, 0.95] for the risk of minor bleed. A + T was found to have the highest RP and the lowest mean value in terms of ACM, MI, and stroke. No significant difference was found between monotherapy or dual-antiplatelet therapy for the major bleeding risk safety outcome, however DAPT was found to have a significantly higher rate of minor bleeding complications post-CABG. DAPT should be considered as the antiplatelet modality of choice post-CABG.


Expression of the mammalian peptide hormone obestatin in Trichoderma reesei.
The filamentous fungus Trichoderma reesei is an expression host widely exploited for the production of recombinant proteins. However, its capacity for expressing small peptides (<20 kDa) has remained largely uncharted to date. In this work, we have produced the hormone peptide obestatin fused to the hydrophobin I tag (Obe-HFBI), using the T. reesei cellobiohydrolase I core (CBHI) or xylanase 2 (XYN2) pro-region as a carrier and the cbh1 promoter for gene expression, in high protein-low protease producing mutant strains T. reesei Rut-C30 and HEPI. The yield of obestatin was improved from about 300 ng/ml to up to 5.5 μg/ml through adaptive laboratory evolution and modifications to the cultivation strategy, which included adjustments of the type and ratio of carbon and nitrogen sources used in the medium. The successful expression of Obe-HFBI demonstrated the potential of T. reesei as an expression host for small peptides and further enhancement of the recombinant yield through modification of culture conditions.


Effects of stimulus contrast on binocular VER.
The effect of stimulus contrast changes on the binocular visual evoked response (VER) was investigated using pattern reversal VER, dichoptic stimulation, and Fast Fourier Transform. When the stimulus contrast was changed binocularly, the monocular components increased as the contrast increased. The binocular component first appeared at the level of 10% contrast; its magnitude was stable under all recording conditions. When the stimulus contrast was changed monocularly, the eye that received the higher contrast stimulation showed more power in the power spectrum. The binocular component appeared even when the difference in contrast between the two eyes was large. The magnitude of the binocular component was stable under all recording conditions. We concluded that the binocular and the monocular components differ in the magnitude of their responses to a contrast change and speculate that the pathways responsible for the two components are not identical.


Male and female physical intimate partner violence and socio-economic position: a cross-sectional international multicentre study in Europe.
This work explores the association between socio-economic position (SEP) and intimate partner violence (IPV) considering the perspectives of men and women as victims, perpetrators and as both (bidirectional). Cross-sectional international multicentre study. A sample of 3496 men and women, (aged 18-64 years), randomly selected from the general population of residents from six European cities was assessed: Athens; Budapest; London; Östersund; Porto; and Stuttgart. Their education (primary, secondary and university), occupation (upper white collar, lower white collar and blue collar) and unemployment duration (never, ≤12 months and >12 months) were considered as SEP indicators and physical IPV was measured with the Revised Conflict Tactics Scales. Past year physical IPV was declared by 17.7% of women (3.5% victims, 4.2% perpetrators and 10.0% bidirectional) and 19.8% of men (4.1% victims, 3.8% perpetrators and 11.9% bidirectional). Low educational level (primary vs university) was associated with female victimisation (adjusted odds ratio, 95% confidence interval: 3.2; 1.3-8.0) and with female bidirectional IPV (4.1, 2.4-7.1). Blue collar occupation (vs upper white) was associated with female victimisation (2.1, 1.1-4.0), female perpetration (3.0, 1.3-6.8) and female bidirectional IPV (4.0, 2.3-7.0). Unemployment duration was associated with male perpetration (>12 months of unemployment vs never unemployed: 3.8; 1.7-8.7) and with bidirectional IPV in both sex (women: 1.8, 1.2-2.7; men: 1.7, 1.0-2.8). In these European centres, physical IPV was associated with a disadvantaged SEP. A consistent socio-economic gradient was observed in female bidirectional involvement, but victims or perpetrators-only presented gender specificities according to levels of education, occupation differentiation and unemployment duration potentially useful for designing interventions.


APE2 is required for ATR-Chk1 checkpoint activation in response to oxidative stress.
The base excision repair pathway is largely responsible for the repair of oxidative stress-induced DNA damage. However, it remains unclear how the DNA damage checkpoint is activated by oxidative stress at the molecular level. Here, we provide evidence showing that hydrogen peroxide (H2O2) triggers checkpoint kinase 1 (Chk1) phosphorylation in an ATR [ataxia-telangiectasia mutated (ATM) and Rad3-related]-dependent but ATM-independent manner in Xenopus egg extracts. A base excision repair protein, Apurinic/apyrimidinic (AP) endonuclease 2 (APE2, APN2, or APEX2), is required for the generation of replication protein A (RPA)-bound single-stranded DNA, the recruitment of a checkpoint protein complex [ATR, ATR-interacting protein (ATRIP), and Rad9] to damage sites, and H2O2-induced Chk1 phosphorylation. A conserved proliferating cell nuclear antigen interaction protein box of APE2 is important for the recruitment of APE2 to H2O2-damaged chromatin. APE2 3'-phosphodiesterase and 3'-5' exonuclease activity is essential for single-stranded DNA generation in the 3'-5' direction from single-stranded breaks, referred to as single-stranded break end resection. In addition, APE2 associates with Chk1, and a serine residue (S86) in the Chk1-binding motif of APE2 is essential for Chk1 phosphorylation, indicating a Claspin-like but distinct role for APE2 in ATR-Chk1 signaling. Our data indicate that APE2 plays a vital and previously unexpected role in ATR-Chk1 checkpoint signaling in response to oxidative stress. Thus, our findings shed light on a distinct mechanism of how an ATR-Chk1-dependent DNA damage checkpoint is mediated by APE2 in the oxidative stress response.


An open canvas--2D materials with defects, disorder, and functionality.
CONSPECTUS: While some exceptional properties are unique to graphene only (its signature Dirac-cone gapless dispersion, carrier mobility, record strength), other features are common to other two-dimensional materials. The broader family "beyond graphene" offers greater choices to be explored and tailored for various applications. Transition metal dichalcogenides (TMDCs), hexagonal boron nitride (h-BN), and 2D layers of pure elements, like phosphorus or boron, can complement or even surpass graphene in many ways and uses, ranging from electronics and optoelectronics to catalysis and energy storage. Their availability greatly relies on chemical vapor deposition growth of large samples, which are highly polycrystalline and include interfaces such as edges, heterostructures, and grain boundaries, as well as dislocations and point defects. These imperfections do not always degrade the material properties, but they often bring new physics and even useful functionality. It turns particularly interesting in combination with the sheer openness of all 2D sheets, fully exposed to the environment, which, as we show herein, can change and tune the defect structures and consequently all their qualities, from electronic levels, conductivity, magnetism, and optics to structural mobility of dislocations and catalytic activities. In this Account, we review our progress in understanding of various defects. We begin by expressing the energy of an arbitrary graphene edge analytically, so that the environment is regarded by "chemical phase shift". This has profound implications for graphene and carbon nanotube growth. Generalization of this equation to heteroelemental BN gives a method to determine the energy for arbitrary edges of BN, depending on the partial chemical potentials. This facilitates the tuning of the morphology and electronic and magnetic properties of pure BN or hybrid BN|C systems. Applying a similar method to three-atomic-layer TMDCs reveals more diverse edge structures for thermodynamically stable flakes. Moreover, CVD samples show new types of edge reconstruction, providing insight into the nonequilibrium growth process. Combining dislocation theory with first-principles computations, we could predict the dislocation cores for BN and TMDC and reveal their variable chemical makeup. This lays the foundation for the unique sensitivity to ambient conditions. For example, partial occupation of the defect states for dislocations in TMDCs renders them intrinsically magnetic. The exchange coupling between electrons from neighboring dislocations in grain boundaries further makes them half-metallic, which may find its applications in spintronics. Finally, brief discussion of monoelemental 2D-layer phosphorus and especially the structures and growth routes of 2D boron shows how theoretical assessment can help the quest for new synthetic routes.


Clival fractures in a level I trauma center.
Clival fracture (CF) is rare among head traumas. The aim of this study was to explore how radiological features observed in CF reflect the clinical picture and mechanism of injury in such cases. Radiological data for patients with skull base fracture admitted to the Montreal General Hospital between February 2002 and October 2012 were obtained from the Quebec Trauma Registry and reviewed for CF. Identified CF was categorized by orientation and quality. Injury mechanism, clinical presentation, and follow-up outcome were obtained through retrospective chart review. Of the 1738 patients with skull base fractures, 65 exhibited CF, representing 1.2% of the 5416 patients with traumatic brain injuries admitted during the period studied. Thirty-nine (60%) of the 65 CFs were obliquely oriented, 17 (26.2%) were longitudinal, and 9 (14%) were transverse. Twenty-nine (45%) of the 65 patients demonstrated linear fracture, 17 (26%) hairline, 10 (15%) diastatic, and 9 (14%) displaced. Cranial nerve deficits and vascular injury occurred in 13.8% and 7.7% of cases, respectively. Twenty-five patients (38.5%) died in hospital. The long-term Extended Glasgow Outcome Scale score was significantly lower in transverse compared with longitudinal and oblique fractures (p = 0.03 and 0.03, respectively) and lower in diastatic compared with displaced fractures (p = 0.05). This study provides information on the largest CF population studied to date, expands the current CF classification to include fracture quality as well as orientation, and underscores the existence of significant differences in pathogenesis and clinical presentation of CF subtypes.


m6A demethylase ALKBH5 suppression contributes to esophageal squamous cell carcinoma progression.
Esophageal squamous cell carcinoma (ESCC) is a highly malignant gastrointestinal cancer with a high recurrence rate and poor prognosis. Although N6-methyladenosine (m6A), the most abundant epitranscriptomic modification of mRNAs, has been implicated in several cancers, little is known about its participation in ESCC progression. We found reduced expression of ALKBH5, an m6A demethylase, in ESCC tissue specimens with a more pronounced effect in T3-T4, N1-N3, clinical stages III-IV, and histological grade III tumors, suggesting its involvement in advanced stages of ESCC. Exogenous expression of ALKBH5 inhibited the in vitro proliferation of ESCC cells, whereas depletion of endogenous ALKBH5 markedly enhanced ESCC cell proliferation in vitro. This suggests ALKBH5 exerts anti-proliferative effects on ESCC growth. Furthermore, ALKBH5 overexpression suppressed tumor growth of Eca-109 cells in nude mice; conversely, depletion of endogenous ALKBH5 accelerated tumor growth of TE-13 cells in vivo. The growth-inhibitory effects of ALKBH5 overexpression are partly attributed to a G1-phase arrest. In addition, ALKBH5 overexpression reduced the in vitro migration and invasion of ESCC cells. Altogether, our findings demonstrate that the loss of ALKBH5 expression contributes to ESCC malignancy.


Radiotherapy for painful bone metastases from hepatocellular carcinoma.
Painful bone metastases are one of the distressing clinical situations in patients with hepatocellular carcinoma (HCC). The purpose of the study was to evaluate the palliative effect of radiotherapy for painful bone metastases from HCC. A retrospective analysis was performed upon 51 patients, who were treated with radiotherapy for painful bone metastases from HCC at 77 sites, from January 1991 to June 2000. Twenty patients (39%) presented synchronous metastases, and the remaining 31 patients (61%) showed metachronous metastases. In addition to bone pain, neurologic symptoms and a palpable mass were presented in 13 (25%) and seven (13%) patients, respectively. Twenty-one patients (41%) had a solitary metastasis and the most frequent site was the vertebra. The total dose of radiotherapy ranged from 12.5 to 50 Gy (median 30 Gy). On the Wisconsin Brief Pain Questionnaire, relief of pain was achieved at 56 sites (73%). The overall survival rate at 2 years was 4% and the median survival time was 5 months. Univariate analysis showed that tumor stage within the liver and the presence of metastases to organs, other than bones, were significant prognostic factors for survival (P < 0.05). In conclusion, radiotherapy provides effective palliation for patients with painful bone metastases from HCC during the substantial median survival time.


Operationalizing Stakeholder Engagement Through the Stakeholder-Centric Engagement Charter (SCEC).
There is a need for clear strategies and procedures to operationalize stakeholder engagement in research studies. Clear guidelines that promote shared leadership among study investigators and research stakeholders are important for inclusive and sustainable partnerships. Such guidelines may take the form of a governance charter and can be a means for encouraging the participation and inclusion of stakeholders who may have little to no experience with research or are otherwise underrepresented in research. This perspective article presents the Stakeholder-Centric Engagement Charter (SCEC), one effort to operationalize a stakeholder engagement approach between researchers and an advisory committee as guided by the Patient-Centered Outcomes Research Institute's (PCORI) Research Engagement Principles (i.e., reciprocal relationships, partnerships, co-learning, transparency-honesty-trust). Building on the SCEC can help future investigators develop a study-specific, dynamic, governance document outlining advisory committee and research team preferences in areas such as role expectations, study governance, and decision-making procedures.


Novel Insights into the Interplay between Apoptosis and Autophagy.
For several decades, apoptosis has taken center stage as the principal mechanism of programmed cell death (type I cell death) in mammalian tissues. Autophagic cell death (type II) is characterized by the massive accumulation of autophagic vacuoles in the cytoplasm of cells. The autophagic process is activated as an adaptive response to a variety of extracellular and intracellular stresses, including nutrient deprivation, hormonal or therapeutic treatment, pathogenic infection, aggregated and misfolded proteins, and damaged organelles. Increasing evidence indicates that autophagy is associated with a number of pathological processes, including cancer. The regulation of autophagy in cancer cells is complex since it can enhance cancer cell survival in response to certain stresses, while it can also act to suppress the initiation of cancer growth. This paper focused on recent advances regarding autophagy in cancer and the techniques currently available to manipulate autophagy.


Childhood obesity treatment: targeting parents exclusively v. parents and children.
There is a consensus that interventions to prevent and treat childhood obesity should involve the family; however, the extent of the child's involvement has received little attention. The goal of the present study was to evaluate the relative efficacy of treating childhood obesity via a family-based health-centred intervention, targeting parents alone v. parents and obese children together. Thirty-two families with obese children of 6-11 years of age were randomised into groups, in which participants were provided for 6 months a comprehensive educational and behavioural programme for a healthy lifestyle. These groups differed in their main agent of change: parents-only v. the parents and the obese child. In both groups, parents were encouraged to foster authoritative parenting styles (parents are both firm and supportive; assume a leadership role in the environmental change with appropriate granting of child's autonomy). Only the intervention aimed at parents-only resulted in a significant reduction in the percentage overweight at the end of the programme (P=0.02) as well as at the 1-year follow-up meeting. The differences between groups at both times were significant (P<0.05). A greater reduction in food stimuli in the home (P<0.05) was noted in the parents-only group. In both groups, the parents' weight status did not change. Regression analysis shows that the level of attendance in sessions explained 28 % of the variability in the children's weight status change, the treatment group explained another 10 %, and the improvement in the obesogenic load explained 11 % of the variability. These results suggest that omitting the obese child from active participation in the health-centred programme may be beneficial for weight loss and for the promotion of a healthy lifestyle among obese children.


GAP-43, aFGF, CCK and alpha- and beta-CGRP in rat spinal motoneurons subjected to axotomy and/or dorsal root severance.
The mRNA levels for growth-associated protein 43 (GAP-43), acidic fibroblast growth factor (aFGF), alpha- and beta-calcitonin gene-related peptide (CGRP), cholecystokinin (CCK) and choline acetyltransferase (ChAT) in rat lumbar spinal motoneurons were studied by in situ hybridization 1, 5 and 21 days and 20 weeks following unilateral peripheral nerve sectioning, ventral rhizotomy or dorsal rhizotomy. Furthermore, CGRP- and aFGF-like immunoreactivities in the ventral horn were studied using immunohistochemistry. One to 21 days after axotomy, GAP-43 and alpha-CGRP mRNAs increased in lesioned motoneurons, while the aFGF mRNA levels were marginally higher in motoneurons on the lesion side as compared to the control side. beta-CGRP, CCK and ChAT mRNA levels, on the other hand, decreased during the short-term response (1-21 days) to axotomy. After ventral rhizotomy, but not peripheral axotomy, there was complete disappearance of aFGF-like immunoreactivity in the ventral root proximal to the lesion. In animals subjected to long-term survival (20 weeks) after peripheral axotomy, the expression of all studied substances had returned to normal levels. Unilateral dorsal rhizotomy did not induce any substantial short- or long-term shifts in the cellular expression of the GAP-43, aFGF, CGRP and CCK peptides or their mRNAs in motoneurons of lesioned segments. These results indicate that peptides/proteins in motoneurons are expressed differentially after axotomy. Whereas alpha-CGRP and GAP-43 are up-regulated, CCK and beta-CGRP become down-regulated and aFGF is largely unaffected.


[Lysosomal hydrolases of various rat organs in the process of cell nutrition].
Activity of lysosomal proteinases cathepsins A, B, C, D, H and L was studied in liver, kidney, spleen, thymus, heart, skeletal muscle and brain tissues of rats, maintained on full value diet, on a diet free of proteins and on the diet enriched with methionine. Distinctly dissimilar reactions of thiol-dependent proteinases was detected in liver, heart tissues and skeletal muscles as compared with the reaction of cathepsins A and D; activity of cathepsins B, H and L was decreased 1.5-2-fold in the animal groups maintained on diets both free of proteins and enriched with methionine. The data obtained suggest that concentration of endogenous inhibitors of lysosomal thiol-dependent proteinases rather than content of SH-groups in tissues was responsible for alterations of their activity under conditions of protein starvation.


Comparison of guided bone formation from periosteum and muscle fascia.
Muscle fascia and periosteum have been used clinically to guide prefabrication of vascularized bone flaps for reconstruction of complex three-dimensional tissues. Although it seems that both locations have the capacity to generate vascularized bone, there have been no studies that directly compare different implantation sites. The authors performed a rigorous, quantitative, histomorphometric comparison of bone prefabrication in a large-animal model comparing graft implanted against muscle fascia and periosteum. Twenty skeletally mature domestic sheep were implanted with rectangular chambers containing equal weights of morcellized bone graft. Two chambers were implanted into each sheep, one with the open face apposed to the cambium layer of the rib periosteum and the other with the open face apposed to the fascia of the latissimus dorsi muscle. Animals were euthanized at 3, 6, 9, 12, and 24 weeks and the chambers were harvested. Tissue inside the chambers was analyzed for shape conformation to chamber geometry, gross tissue volume, and bone histomorphology. There were no differences in volume or shape of tissue formed in the chambers. However, chambers in contact with fascia consisted almost entirely of fibrovascular tissue, with progressive resorption of the morcellized bone graft and little evidence of new bone. Chambers in contact with periosteum showed active endochondral, direct, and appositional bone formation over time, with increasing calcified tissue area and new bone formation. Both periosteum and muscle fascia were able to vascularize bone grafts, but bone formation was higher in the periosteum. The periosteum appears to be a more suitable foundation from which to promote flap prefabrication.


Incidence and risk factors of pressure ulcers in seven neonatal intensive care units in Japan: a multisite prospective cohort study.
This study aims to clarify (i) the incidence of pressure ulcers in neonates admitted to the neonatal intensive care units (NICUs) and (ii) risk factors of pressure ulcer development. All infants admitted to the NICU and kept in incubators from seven hospitals during the study period were recruited to the study. Each infant was given skin examination every day by nurses, and risk factors were collected three times a week by one researcher. The incidence of the pressure ulcers was calculated, and the risk factors for pressure ulcers were determined by using univariate and multivariate analysis. Eighty-one infants were involved in the study. A total of 14 pressure ulcers occurred in 13 infants during the 11-month study period, the incidence was 0·01 persons per day and cumulative incidence rate was 16.0%. Seven (50·0%) of 14 pressure ulcers were located on the nose. Multivariate analysis identified the following risk factors: skin texture (Dubowitz neonatal maturation assessment scale: skin texture score of 1 point or lower) [odds ratio 7·6; 95% confidence interval (CI) 1·58 -36·71, P = 0·012] and endotracheal intubation usage (odds ratio 4·0; 95% CI 1·04-15·42, P = 0·042).


Vaginal and Uterine Bacterial Communities in Postpartum Lactating Cows.
Reproductive inefficiency in cattle has major impacts on overall productivity of cattle operations, increasing cost of production, and impacting the sustainability of the cattle enterprise. Decreased reproductive success and associated disease states have been correlated with the presence of specific microbes and microbial community profiles, yet details of the relationship between microbial communities and host physiology are not well known. The present study profiles and compares the microbial communities in the bovine uterus and vagina using 16S rRNA sequencing of the V1-V3 hypervariable region at the time of artificial insemination. Significant differences (p < 0.05) between the vaginal and uterine communities were observed at the level of α-diversity metrics, including Chao1, Shannon's Diversity Index, and observed OTU. Greater clustering of vaginal OTU was apparent in principal coordinate analysis compared to uterine OTU, despite greater diversity in the vaginal community in both weighted and unweighted UniFrac distance matrices (p < 0.05). There was a significantly greater relative abundance of unassigned taxa in the uterus (p = 0.008), otherwise there were few differences between the overall community profiles. Both vaginal and uterine communities were dominated by Firmicutes, although the relative abundance of rRNA sequences corresponding to species in this phylum was significantly (p = 0.007) lower in the uterine community. Additional differences were observed at the genus level, specifically in abundances within Clostridium (p = 0.009), Anaerofustis (p = 0.018), Atopobium (p = 0.035), Oscillospira (p = 0.035), 5-7N15 (p = 0.035), Mycoplasma (p = 0.035), Odoribacter (p = 0.042), and within the families Clostridiaceae (p = 0.006), Alcaligenaceae (p = 0.021), and Ruminococcaceae (p = 0.021). Overall, the comparison revealed differences and commonalities among bovine reproductive organs, which may be influenced by host physiology. The increased abundance of unassigned taxa found in the uterus may play a significant biological role in the reproductive status of the animal. The study represents an initial dataset for comparing bacterial communities prior to establishment of pregnancy.


Heterozygous DHTKD1 Variants in Two European Cohorts of Amyotrophic Lateral Sclerosis Patients.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by progressive upper and lower motor neuron (LMN) loss. As ALS and other neurodegenerative diseases share genetic risk factors, we performed whole-exome sequencing in ALS patients focusing our analysis on genes implicated in neurodegeneration. Thus, variants in the DHTKD1 gene encoding dehydrogenase E1 and transketolase domain containing 1 previously linked to 2-aminoadipic and 2-oxoadipic aciduria, Charcot-Marie-Tooth (CMT) disease type 2, and spinal muscular atrophy (SMA) were identified. In two independent European ALS cohorts (n = 643 cases), 10 sporadic cases of 225 (4.4%) predominantly sporadic patients of cohort 1, and 12 familial ALS patients of 418 (2.9%) ALS families of cohort 2 harbored 14 different rare heterozygous DHTKD1 variants predicted to be deleterious. Four DHTKD1 variants were previously described pathogenic variants, seven were recurrent, and eight were located in the E1_dh dehydrogenase domain. Nonsense variants located in the E1_dh domain were significantly more prevalent in ALS patients versus controls. The phenotype of ALS patients carrying DHTKD1 variants partially overlapped with CMT and SMA by presence of sensory impairment and a higher frequency of LMN-predominant cases. Our results argue towards rare heterozygous DHTKD1 variants as potential contributors to ALS phenotype and, possibly, pathogenesis.


Regular physical exercise and bone mineral density: a four-year controlled randomized trial in middle-aged men. The DNASCO study.
The aim of the study was to investigate the effects of regular aerobic exercise training on bone mineral density (BMD) in middle-aged men. A population based sample of 140 men (53-62 years) was randomly assigned into the exercise and reference groups. BMD and apparent volumetric BMD (BMDvol) of the proximal femur and lumbar spine (dual-energy X-ray absorptiometry, DXA) and anthropomorphic measurements were performed at the randomization and 2 and up to 4 years later. The participation rate was 97% and 94% at the second and third BMD measurements, respectively. As another indication of excellent adherence and compliance, the cardiorespiratory fitness (aerobic threshold) increased by 13% in the exercise group. The 2% decrease in the reference group is regarded as an age-related change in cardiorespiratory fitness. Regardless of the group, there was no association between the increase in aerobic threshold and change in BMD. In the entire group, age-related bone loss was seen in the femoral neck BMD and BMDvol (p < 0.01). BMD and BMDvol values increased with age in L2-L4 (p < 0.004). An increased rate of bone loss at the femoral neck was observed in men with a low energy-adjusted calcium intake (p = 0.003). Men who increased their alcohol intake during the intervention showed a decrease in the rate of bone loss at the femoral neck (p = 0.040). A decrease in body height associated with decreased total femoral BMD (r = 0.19, p = 0.04) and the change in body height was a predictor of bone loss in the femoral neck (beta = 0.201). Long-term regular aerobic physical activity in middle-aged men had no effect on the age-related loss of femoral BMD. On the other hand, possible structural alterations, which are also essential for the mechanical strength of bone, can not be detected by the DXA measurements used in this study. The increase seen in lumbar BMD reflects age-related changes in the spine, thus making it an unreliable site for BMD follow-up in men.


Influence of genomic ancestry on the distribution of SLCO1B1, SLCO1B3 and ABCB1 gene polymorphisms among Brazilians.
The frequency distribution of SNPs and haplotypes in the ABCB1, SLCO1B1 and SLCO1B3 genes varies largely among continental populations. This variation can lead to biases in pharmacogenetic studies conducted in admixed populations such as those from Brazil and other Latin American countries. The aim of this study was to evaluate the influence of self-reported colour, geographical origin and genomic ancestry on distributions of the ABCB1, SLCO1B1 and SLCO1B3 polymorphisms and derived haplotypes in admixed Brazilian populations. A total of 1039 healthy adults from the north, north-east, south-east and south of Brazil were recruited for this investigation. The c.388A>G (rs2306283), c.463C>A (rs11045819) and c.521T>C (rs4149056) SNPs in the SLCO1B1 gene and c.334T>G (rs4149117) and c.699G>A (rs7311358) SNPs in the SLCO1B3 gene were determined by Taqman 5'-nuclease assays. The ABCB1 c.1236C>T (rs1128503), c.2677G>T/A (rs2032582) and c.3435C>T (rs1045642) polymorphisms were genotyped using a previously described single-base extension/termination method. The results showed that genotype and haplotype distributions are highly variable among populations of the same self-reported colour and geographical region. However, genomic ancestry showed that these associations are better explained by a continuous variable. The influence of ancestry on the distribution of alleles and haplotype frequencies was more evident in variants with large differences in allele frequencies between European and African populations. Design and interpretation of pharmacogenetic studies using these transporter genes should include genomic controls to avoid spurious conclusions based on improper matching of study cohorts from Brazilian populations and other highly admixed populations.


Hazardous drinking among restaurant workers.
We address three research questions pertaining to Swedish restaurant workers: (i) What is the prevalence of hazardous drinking? (ii) How is the consumption of alcohol distributed? (iii) Does the prevention paradox apply? Data were collected by administering the Alcohol Use Disorders Identification Test (AUDIT) among restaurant workers who attended a 2-day Responsible Beverage Service training in Stockholm during the period from October 2008 to December 2009. The control group comprised a sample representative of the general Swedish population. We restricted the analyses to the age span 18-59 years, which yielded a sample size of 579 for restaurant workers and 434 for the general population. The prevalence of hazardous drinking as measured by AUDIT (8+ for men and 6+ for women) was markedly higher among restaurant workers than in the general population. The difference was especially pronounced among females below 30 years of age. We found no difference between restaurant workers and the general population in the distribution of alcohol consumption. About 76% of the drinking problems were found in the lower part of the consumption distribution (bottom 88%), which supports the prevention paradox. Restaurant workers comprise a high-risk group with respect to drinking.


Short-term results of arthroscopic treatment of osteochondritis dissecans in skeletally immature patients.
Osteochondritis dissecans (OCD) of the capitellum occurs in skeletally immature athletes, and most likely results from repetitive trauma during overhead activities. Treatment may consist of activity modifications, internal fixation, abrasion chondroplasty, microfracture, antegrade drilling, retrograde drilling, osteochondral autograft or allograft implantation, chondrocyte implantation, and rib autograft through arthroscopy or arthrotomy. One treatment modality has not been proven to be clearly more effective than the others. This study was undertaken to evaluate our treatment regimen that utilized arthroscopic-assisted treatments of capitellar OCD, including removal of loose bodies, antegrade or retrograde drilling, and chondroplasty in the pediatric population. All patients treated arthroscopically for a diagnosis of capitellar OCD over a 5-year period, were retrospectively reviewed. All were asked to return for follow-up questionnaire and radiographs. Exclusion criteria included those lost to follow-up. Demographics were recorded and range of motion was evaluated preoperatively and at most recent follow-up for flexion, extension, supination, and pronation. Preoperative and the most recent anterior/posterior and lateral radiographs of the elbow were reviewed. A 200-point elbow rating scale was used to assess patient outcomes. The arthroscopic appearance of the lesion was graded. Changes in preoperative to postoperative range of motion and size of lesion were compared using repeated measures analysis of variation. There were 13 elbows in 12 patients; only 3 of the injuries occurred acutely. Mean age at the time of surgery was 13.1±1.07 (range, 10.8 to 14.6 y). Mean follow-up was 23.4±16.7 months (range, 2 to 60 mo). There were 3 grade I lesions, 2 grade II lesions, 1 grade IV lesion, and 7 grade V lesions. Seven of the lesions underwent transhumeral drilling, 2 transarticular drilling, 3 loose body removals, and 2 had only debridement. There were no postoperative infections or neurovascular injuries. Three of the elbows (20%) required eventual arthrotomy at a mean of 27.9 months after the index procedure. At final follow-up, 67% reported no pain and 33% reported occasional pain. No patients reported any swelling. Eighty-three percent reported no locking or catching and 17% reported occasional locking or catching. No patients reported any activity restrictions. The mean subjective score was 96±6 of 100 and the mean objective score was 100 of 100. Mean extension improved significantly from -17 to -7 degrees (P<0.001). Treatment of children with OCD lesions of the capitellum with arthroscopic-assisted debridement and fenestration of the sclerotic rim (trans-humeral if overlaying cartilage is intact), plus fixation of the overlaying cartilage if not securely attached to the subchondral bone permits the return to physical activity, but may not allow return to the injury-inducing sport. Our short-term outcomes obtained using this regimen found this technique to be safe and reliable, but other interventions may be required if continued disability persists.


Mechanisms of impaired potassium handling with dual renin-angiotensin-aldosterone blockade in chronic kidney disease.
The combination of an aldosterone receptor antagonist added to an angiotensin-converting enzyme inhibitor has been demonstrated to reduce cardiovascular and renal end points in hypertensive humans but can produce hyperkalemia in the common clinical setting of impaired renal function. We investigated the effects of dual therapy on acute and chronic potassium handling in hypertensive humans with renal impairment by conducting a randomized crossover clinical trial of 4 weeks of 40 mg lisinopril/25 mg spironolactone versus placebo in 18 participants with a glomerular filtration rate of 25 to 65 mL/min. Study end points, following an established protocol, were hourly determinations of dynamic renal potassium excretion (mmol/h) and serum potassium (mmol/L) after 35 mmol oral potassium challenge in addition to ambulatory potassium concentration. After 4 weeks, ambulatory potassium concentration was 4.87 mmol/L with lisinopril/spironolactone versus 4.37 with placebo (P<0.001). Lisinopril/spironolactone produced only a modest 0.44 mmol/h reduction in stimulated potassium excretion (P=0.03) but a substantial 0.67 mmol/L increase in serum potassium (P<0.001) in response to 35 mmol potassium; these findings are consistent with impaired extrarenal/transcellular potassium disposition. We found the increase in serum potassium after an oral potassium challenge to be a strong predictor of the increase in ambulatory potassium with lisinopril/spironolactone. Our study suggests that dual renin-angiotensin-aldosterone blockade may impair extrarenal/transcellular potassium disposition in addition to reducing potassium excretion in humans with renal impairment, and that acute changes in dynamic potassium handling are predictive of chronic changes in ambulatory potassium concentration with dual renin-angiotensin-aldosterone blockade.


The clinical usefulness of serum prostate specific antigen after hormonal therapy of metastatic prostate cancer.
We longitudinally followed serum prostate specific antigen (PSA) levels in 48 patients who were treated with either orchiectomy, monthly luteinizing hormone-releasing hormone injection or continuous diethylstilbestrol for stage D2 prostate adenocarcinoma and achieved an objective response. Of the patients 34 had clinical evidence of disease progression (median remission duration 19 months). Median length of followup for the 14 patients who remained in remission was 42 months. Pretreatment performance status, pretreatment extent of metastases as measured by a bone scan and post-treatment nadir PSA level were univariately correlated with remission duration. After adjustment for the 2 former pretreatment variables, a highly significant independent effect of the nadir PSA level on remission duration persisted. Patients whose post-treatment nadir PSA level decreased below 4 ng./ml. had a significantly longer remission duration than those whose nadir PSA remained elevated (median 42 versus 10 months, p less than 0.0001). No cases were observed to progress (as defined by our criteria independent of PSA level) while the serial post-treatment PSA levels continued to decrease or remained at a plateau after reaching the nadir. The time at which the PSA began to increase once the nadir was reached predated objective evidence of progression in all patients except 2 in whom the 2 events occurred simultaneously (mean lead time 7.3 +/- 5.0 months). We conclude that following serial PSA levels in patients treated with androgen ablation for metastatic prostate cancer can aid in distinguishing favorable from nonfavorable responders early in the course of therapy and greatly assist in monitoring for progression.


Endothelial ether lipids link the vasculature to blood pressure, behavior, and neurodegeneration.
Vascular disease contributes to neurodegeneration, which is associated with decreased blood pressure in older humans. Plasmalogens, ether phospholipids produced by peroxisomes, are decreased in Alzheimer's disease, Parkinson's disease, and other neurodegenerative disorders. However, the mechanistic links between ether phospholipids, blood pressure, and neurodegeneration are not fully understood. Here, we show that endothelium-derived ether phospholipids affect blood pressure, behavior, and neurodegeneration in mice. In young adult mice, inducible endothelial-specific disruption of PexRAP, a peroxisomal enzyme required for ether lipid synthesis, unexpectedly decreased circulating plasmalogens. PexRAP endothelial knockout (PEKO) mice responded normally to hindlimb ischemia but had lower blood pressure and increased plasma renin activity. In PEKO as compared with control mice, tyrosine hydroxylase was decreased in the locus coeruleus, which maintains blood pressure and arousal. PEKO mice moved less, slept more, and had impaired attention to and recall of environmental events as well as mild spatial memory deficits. In PEKO hippocampus, gliosis was increased, and a plasmalogen associated with memory was decreased. Despite lower blood pressure, PEKO mice had generally normal homotopic functional connectivity by optical neuroimaging of the cerebral cortex. Decreased glycogen synthase kinase-3 phosphorylation, a marker of neurodegeneration, was detected in PEKO cerebral cortex. In a co-culture system, PexRAP knockdown in brain endothelial cells decreased glycogen synthase kinase-3 phosphorylation in co-cultured astrocytes that was rescued by incubation with the ether lipid alkylglycerol. Taken together, our findings suggest that endothelium-derived ether lipids mediate several biological processes and may also confer neuroprotection in mice.


Ambulatory outcome of children with myelomeningocele: effect of lower-extremity muscle strength.
The relationship between patterns of strength and mobility was studied in 291 children with myelomeningocele, graded as community ambulators, partial (household) ambulators and nonambulators. Iliopsoas strength was found to be the best predictor of ambulation, with the quadriceps, anterior tibialis and glutei also contributing significantly. Grade 0 to 3 iliopsoas strength was always associated with partial or complete reliance on a wheelchair. No patient with grade 4 to 5 iliopsoas and quadriceps function relied completely on wheelchairs and the majority were community ambulators. Grade 4 to 5 gluteal and anterior tibialis function was associated with community ambulation, without aids or braces. Deterioration in mobility was most common in those with strong iliopsoas/quadriceps and grade 0 to 3 gluteus medius, and was not age-related.


Aetiology, antifungal susceptibility, risk factors and outcome in 201 fungaemic children: data from a 12-year prospective national study from Slovakia.
A total of 201 cases of fungaemia in children in a 12-year national survey from seven University Paediatric Clinics in Slovakia in 1990-2001 was assessed to determine risk factors, therapy and outcome, and to compare those cases with fungaemia in 130 adult cancer patients studied in a similar survey. Four univariate analyses were performed to assess differences in aetiology, antifungal susceptibility and outcome between fungaemia in neonates and paediatric intensive care unit (ICU) patients as well as between paediatric and adult cancer patients with fungaemia. There was a significant difference in aetiology and antifungal susceptibility between the subgroups of children with fungaemia: 83.3% of neonates versus 40.2% in children with cancer were due to Candida albicans. None of the non-albicans Candida spp. (NAC) in neonates but 23.5% of NAC isolates from children with cancer were resistant to fluconazole. C. albicans caused 144 (71.1%) episodes and NAC 48 (23.7%) episodes. Trichosporon beigelii, Blastoschizomyces (Trichosporon) capitatus, Rhodotorula rubra and Cryptococcus laurentii were found less frequently in neonates than in children with cancer (18.8%). There were not many differences in risk factors between paediatric fungaemia and adult cancer fungaemia except C. albicans aetiology, corticosteroid use in therapy, breakthrough fungaemia after ketoconazole prophylaxis and meningitis as a complication, which were observed significantly more frequently among children than in adults, both with cancer and fungaemia. Thirty-three of the paediatric fungaemias were breakthrough cases and appeared frequently in children with cancer. Fifty-one (25.1%) children died with fungaemia (attributable mortality) and 25 (12.7%) due to underlying disease with fungaemia; overall mortality was 37.8% and there was no significant difference in death rates between the subgroups of paediatric patients (neonates, children in ICUs and children with cancer).


Identifying factors associated with hospital readmissions among stroke patients in Taipei.
Hospital readmissions contribute significantly to the cost of medical care, and may reflect unresolved problems at discharge or a lack of resources in post-hospital care. The purpose of this paper is to assess the effects of patient characteristics at discharge, the need for nursing care, discharge planning program, post-hospital care arrangements, and caregiver characteristics on readmissions of stroke patients. Patients discharged from neurological wards in seven hospitals in the Taipei area were recruited into the study. Surveys were conducted before their discharge, and at one month after discharge. Of the 489 patients included in the study, 24.3% were readmitted. After controlling for other variables, factors associated with readmissions were number of limitations in activities of daily living (ADL), first incidence of stroke, the need for wound nursing care, the adoption of a care plan, and the discharge locations. Contrary to expectation, age, length of stay, counseling before discharge, and caregiver burden were not associated with readmissions. The findings of this study indicate that ADL limitation is an effective predictor of readmissions. Increasing home nursing resources to meet the demand for wound nursing care may also be effective in reducing readmissions. Discharging patients into institutions for a short period of time may also prove to be more economically viable due to the reduction in readmissions.


Transgenic mice with increased Cu/Zn-superoxide dismutase activity are resistant to N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity.
Administration of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to mammals causes damage to the nigrostriatal dopaminergic pathway similar to that observed in Parkinson's disease. It has been suggested that the mechanism by which MPTP kills dopamine (DA) neurons involves an energy crisis due to the inhibition of mitochondrial complex I. In addition, superoxide radicals (O2-), generated subsequent to the blockade of mitochondrial complex I, may also be involved in MPTP-induced neurotoxicity. Superoxide dismutase (SOD) is a scavenger enzyme that protects cells from the hazard of O2- radicals. To evaluate further the role of O2- radical in MPTP-induced toxicity, we tested the effects of MPTP in transgenic mice with increased SOD activity. In nontransgenic littermates with normal SOD activity, MPTP injection causes a marked reduction in striatal levels of DA and its metabolites as well as in striatal and nigral 3H-DA uptake; these findings are consistent with a loss in dopaminergic neurons. In contrast, in transgenic mice with increased SOD activity, MPTP injection does not cause any significant changes either in levels of DA and metabolites or in 3H-DA uptake. We show that this lack of toxicity is not due to a lower delivery of MPTP to the brain following its intraperitoneal injection, to reduced brain biotransformation of MPTP to N-methyl-4-phenylpyridinium ion (MPP+), to diminished striatal mitochondrial monoamine oxidase B activity, to decreased synaptosomal uptake of MPP+, to lower potency of MPP+ to inhibit the complex I of the mitochondrial electron transport chain, or to faster brain elimination of MPP+. These results suggest that increased SOD activity is, most likely, the protective factor that confers resistance to transgenic mice against MPTP-induced neurotoxicity. Thus, this study provides further evidence that some of the deleterious effects of MPTP may be mediated by O2- radicals. The similarity between the MPTP model and Parkinson's disease further raises the possibility that oxy-radicals may play a significant role in the etiology of this neurodegenerative disorder.


Delivering maternal and neonatal health interventions in conflict settings: a systematic review.
While much progress was made throughout the Millennium Development Goals era in reducing maternal and neonatal mortality, both remain unacceptably high, especially in areas affected by humanitarian crises. While valuable guidance on interventions to improve maternal and neonatal health in both non-crisis and crisis settings exists, guidance on how best to deliver these interventions in crisis settings, and especially in conflict settings, is still limited. This systematic review aimed to synthesise the available literature on the delivery on maternal and neonatal health interventions in conflict settings. We searched MEDLINE, Embase, CINAHL and PsycINFO databases using terms related to conflict, women and children, and maternal and neonatal health. We searched websites of 10 humanitarian organisations for relevant grey literature. Publications reporting on conflict-affected populations in low-income and middle-income countries and describing a maternal or neonatal health intervention delivered during or within 5 years after the end of a conflict were included. Information on population, intervention, and delivery characteristics were extracted and narratively synthesised. Quantitative data on intervention coverage and effectiveness were tabulated but no meta-analysis was undertaken. 115 publications met our eligibility criteria. Intervention delivery was most frequently reported in the sub-Saharan Africa region, and most publications focused on displaced populations based in camps. Reported maternal interventions targeted antenatal, obstetric and postnatal care; neonatal interventions focused mostly on essential newborn care. Most interventions were delivered in hospitals and clinics, by doctors and nurses, and were mostly delivered through non-governmental organisations or the existing healthcare system. Delivery barriers included insecurity, lack of resources and lack of skilled health staff. Multi-stakeholder collaboration, the introduction of new technology or systems innovations, and staff training were delivery facilitators. Reporting of intervention coverage or effectiveness data was limited. The relevant existing literature focuses mostly on maternal health especially around the antenatal period. There is still limited literature on postnatal care in conflict settings and even less on newborn care. In crisis settings, as much as in non-crisis settings, there is a need to focus on the first day of birth for both maternal and neonatal health. There is also a need to do more research on how best to involve community members in the delivery of maternal and neonatal health interventions. CRD42019125221.


Secretion of active human lecithin-cholesterol acyltransferase by insect cells infected with a recombinant baculovirus.
Difficulties in purifying the plasma enzyme lecithin-cholesterol acyltransferase (LCAT) have hampered detailed studies of its (patho)physiological role in lipoprotein metabolism and of structure-function relationships. Potentially, baculovirus-driven expression systems offer a powerful means to produce significant amounts of LCAT. Accordingly, full-length LCAT cDNA was cloned into pVL 1392, a high-level expression derivative of Autographa californica nuclear polyhedrosis virus (AcNPV), and the resultant plasmid was co-transfected into Trichoplusia ni insect cells (High 5 line) with a linearized viral DNA using lipofectin. Such viral DNA had a lethal mutation and grew only when recombined with a pVL1392-type rescue plasmid; cells infected with recombinant Autographa californica LCAT virus changed from a fibroblast-like morphology to rounded, but lacked the polyhedrin occlusion bodies characteristic of wild-type AcNPV infections. Enzymically active recombinant LCAT (rLCAT), sensitive to sulphydryl reagents, was secreted in the late phase of infection (36-48 h) but was absent with wild-type infections. The secreted protein had an apparent molecular mass of 53 kDa by SDS/PAGE, lower than that of native plasma LCAT; it was susceptible to endoglycosidase H digestion and was bound by concanavalin A, suggesting that precursor high-mannose N-glycan chains had not undergone full maturation to complex types. Pretreatment of the cells with tunicamycin to inhibit the first step of N-glycosylation led to intracellular accumulation of immature rLCAT (approximately 46-48 kDa) and a marked reduction in enzyme secreted. We conclude that the baculovirus gene-expression system will permit production of biologically active normal and mutant forms of LCAT protein.


Exciton Coupling in Redox-Active Salen based Self-Assembled Metallacycles.
The incorporation of a redox-active nickel salen complex into supramolecular structures was explored via coordination-driven self-assembly with homobimetallic ruthenium complexes (bridged by oxalato or 5,8-dihydroxy-1,4-naphthoquinato ligands). The self-assembly resulted in the formation of a discrete rectangle using the oxalato complex and either a rectangle or a catenane employing the larger naphthoquinonato complex. The formation of the interlocked self-assembly was determined to be solvent and concentration dependent. The electronic structure and stability of the oxidized metallacycles was probed using electrochemical experiments, UV-Vis-NIR absorption, EPR spectroscopy and DFT calculations, confirming ligand radical formation. Exciton coupling of the intense near-infrared (NIR) ligand radical intervalence charge transfer (IVCT) bands provided further confirmation of the geometric and electronic structures in solution.


Visual acuity norms in pre-school children: the Multi-Ethnic Pediatric Eye Disease Study.
To provide population-based normative data for monocular visual acuity (VA) and interocular differences in VA (IOD) in Black and Hispanic children 30 to 72 months of age without visually significant refractive errors or ophthalmic abnormalities. In a population-based cohort of children in the Multi-Ethnic Pediatric Eye Disease Study, monocular HOTV VA measurements using the Amblyopia Treatment Study protocol were analyzed using continuous and dichotomous outcomes for VA and IOD, after excluding subjects with ophthalmic abnormalities or refractive error. The analysis cohort consisted of 1722 Black and Hispanic children aged 30 to 72 months. Mean logMAR VA improved with age (p < 0.0001) and male gender (p = 0.0008). The proportion of children achieving VA 20/40 or better was associated with age (p < 0.0001), but not ethnicity or gender, and was 81, 94, 99, and virtually 100% in children aged 30 to 35, 36 to 47, 48 to 59, and 60 to 72 months of age, respectively. The most stringent VA threshold that excluded <5% of normal children was 20/63, 20/50, 20/32, and 20/32 for children aged 30 to 35, 36 to 47, 48 to 59, and 60 to 72 months, respectively. Children attending preschool or daycare achieved VA 20/32 more often than those not attending, after age adjustment (p = 0.01), as did children from higher-income families (p = 0.04). There was no association between mean absolute IOD and age (p = 0.45), ethnicity (p = 0.12), or gender (p = 0.19). The proportion of children with an IOD of 0 to 1 lines was higher in males than females (p = 0.02); it did not vary by age (p = 0.06) or ethnicity (p = 0.17). An IOD of 2 or more lines occurred in 6% of normal children. VA test performance in normal pre-school children improves with age. We propose new age-specific thresholds for defining abnormal monocular VA using HOTV optotypes in children between 2 and 5 years of age, for use in screening, clinical practice, and research.