Computed Tomography-Guided Methylene Blue Labeling Prior to Thoracoscopic Resection of Small Deeply Placed Pulmonary Nodules. Do We Really Need Palpation?
Background Video-assisted thoracic surgery (VATS) is widely used for thoracic surgery operations, and day by day it becomes routine for the excision of undetermined pulmonary nodules. However, it is sometimes hard to reach millimetric nodules through a VATS incision. Therefore, some additional techniques were developed to reach such nodules little in size and which are settled on a challenging localization. In the literature, coils, hook wires, methylene blue, lipidol, and barium staining, and also ultrasound guidance were described for this aim. Herein we discuss our experience with CT-guided methylene blue labeling of small, deeply located pulmonary nodules just before VATS excision. Method From April 2013 to October 2016, 11 patients with millimetric pulmonary nodules (average 8, 7 mm) were evaluated in our clinic. For all these patients who had strong predisposing factors for malignancy, an 18F-FDG PET-CT scan was also performed. The patients whose nodules were decided to be excised were consulted the radiology clinic. The favorable patients were taken to CT room 2 hours prior to the operation, and CT-guided methylene blue staining were performed under sterile conditions. Results Mean nodule size of 11 patients was 8.7 mm (6, 2-12). Mean distance from the visceral pleural surface was 12.7 mm (4-29.3). Four of the nodules were located on the left (2 upper lobes, 2 lower lobes), and seven of them were on the right (four lower lobes, two upper lobes, one middle lobe). The maximum standardized uptake values (SUV max) on 18F-FDG PET/CT scan ranged between 0 and 2, 79. Conclusion CT-guided methylene blue staining of millimetric deeply located pulmonary nodules is a safe and feasible technique that helps surgeon find these undetermined nodules by VATS technique without any need of digital palpation.


The ELPA library: scalable parallel eigenvalue solutions for electronic structure theory and computational science.
Obtaining the eigenvalues and eigenvectors of large matrices is a key problem in electronic structure theory and many other areas of computational science. The computational effort formally scales as O(N(3)) with the size of the investigated problem, N (e.g. the electron count in electronic structure theory), and thus often defines the system size limit that practical calculations cannot overcome. In many cases, more than just a small fraction of the possible eigenvalue/eigenvector pairs is needed, so that iterative solution strategies that focus only on a few eigenvalues become ineffective. Likewise, it is not always desirable or practical to circumvent the eigenvalue solution entirely. We here review some current developments regarding dense eigenvalue solvers and then focus on the Eigenvalue soLvers for Petascale Applications (ELPA) library, which facilitates the efficient algebraic solution of symmetric and Hermitian eigenvalue problems for dense matrices that have real-valued and complex-valued matrix entries, respectively, on parallel computer platforms. ELPA addresses standard as well as generalized eigenvalue problems, relying on the well documented matrix layout of the Scalable Linear Algebra PACKage (ScaLAPACK) library but replacing all actual parallel solution steps with subroutines of its own. For these steps, ELPA significantly outperforms the corresponding ScaLAPACK routines and proprietary libraries that implement the ScaLAPACK interface (e.g. Intel's MKL). The most time-critical step is the reduction of the matrix to tridiagonal form and the corresponding backtransformation of the eigenvectors. ELPA offers both a one-step tridiagonalization (successive Householder transformations) and a two-step transformation that is more efficient especially towards larger matrices and larger numbers of CPU cores. ELPA is based on the MPI standard, with an early hybrid MPI-OpenMPI implementation available as well. Scalability beyond 10,000 CPU cores for problem sizes arising in the field of electronic structure theory is demonstrated for current high-performance computer architectures such as Cray or Intel/Infiniband. For a matrix of dimension 260,000, scalability up to 295,000 CPU cores has been shown on BlueGene/P.


On the phylogenetic position of Raphidocystis pallida with some notes on its life cycle.
Raphidocystis pallida, a centrohelid heliozoan with unusually shaped tubular siliceous scales, was reisolated from Jamor river, Portugal, and studied with the use of light and electron microscopy. In enriched cultures, the cells were naked, devoid of siliceous external skeleton with the exception of several scales present in one cell. Instead, such cells were covered with a layer of rod-shaped bacteria. In clonal cultures, the cells gradually acquired siliceous coverings typical for this species and retained them in next generations. Phylogenetic position of R. pallida was clarified with SSU rDNA-based molecular phylogenetics, and its placement within the genus Raphidocystis despite unusual coverings structure was confirmed. The implications of phylogenetic placement of R. pallida and possible origins of the previously undescribed naked form were discussed.


Early Life Exposure to Low Levels of AHR Agonist PCB126 (3,3',4,4',5-Pentachlorobiphenyl) Reprograms Gene Expression in Adult Brain.
Early life exposure to environmental chemicals can have long-term consequences that are not always apparent until later in life. We recently demonstrated that developmental exposure of zebrafish to low, nonembryotoxic levels of 3,3',4,4',5-pentachlorobiphenyl (PCB126) did not affect larval behavior, but caused changes in adult behavior. The objective of this study was to investigate the underlying molecular basis for adult behavioral phenotypes resulting from early life exposure to PCB126. We exposed zebrafish embryos to PCB126 during early development and measured transcriptional profiles in whole embryos, larvae and adult male brains using RNA-sequencing. Early life exposure to 0.3 nM PCB126 induced cyp1a transcript levels in 2-dpf embryos, but not in 5-dpf larvae, suggesting transient activation of aryl hydrocarbon receptor with this treatment. No significant induction of cyp1a was observed in the brains of adults exposed as embryos to PCB126. However, a total of 2209 and 1628 genes were differentially expressed in 0.3 and 1.2 nM PCB126-exposed groups, respectively. KEGG pathway analyses of upregulated genes in the brain suggest enrichment of calcium signaling, MAPK and notch signaling, and lysine degradation pathways. Calcium is an important signaling molecule in the brain and altered calcium homeostasis could affect neurobehavior. The downregulated genes in the brain were enriched with oxidative phosphorylation and various metabolic pathways, suggesting that the metabolic capacity of the brain is impaired. Overall, our results suggest that PCB exposure during sensitive periods of early development alters normal development of the brain by reprogramming gene expression patterns, which may result in alterations in adult behavior.


Zebrafish: Housing and husbandry recommendations.
This article provides recommendations for the care of laboratory zebrafish (Danio rerio) as part of the further implementation of Annex A to the European Convention on the protection of vertebrate animals used for experimental and other scientific purposes, EU Commission Recommendation 2007/526/EC and the fulfilment of Article 33 of EU Directive 2010/63, both concerning the housing and care of experimental animals. The recommendations provide guidance on best practices and ranges of husbandry parameters within which zebrafish welfare, as well as reproducibility of experimental procedures, are assured. Husbandry procedures found today in zebrafish facilities are numerous. While the vast majority of these practices are perfectly acceptable in terms of zebrafish physiology and welfare, the reproducibility of experimental results could be improved by further standardisation of husbandry procedures and exchange of husbandry information between laboratories. Standardisation protocols providing ranges of husbandry parameters are likely to be more successful and appropriate than the implementation of a set of fixed guidance values neglecting the empirically successful daily routines of many facilities and will better reflect the wide range of environmental parameters that characterise the natural habitats occupied by zebrafish. A joint working group on zebrafish housing and husbandry recommendations, with members of the European Society for Fish Models in Biology and Medicine (EUFishBioMed) and of the Federation of European Laboratory Animal Science Associations (FELASA) has been given a mandate to provide guidelines based on a FELASA list of parameters, 'Terms of Reference'.


Relocation of the NIb gene in the tobacco etch potyvirus genome.
Potyviruses express most of their proteins from a long open reading frame that is translated into a large polyprotein processed by three viral proteases. To understand the constraints on potyvirus genome organization, we relocated the viral RNA-dependent RNA polymerase (NIb) cistron to all possible intercistronic positions of the Tobacco etch virus (TEV) polyprotein. Only viruses with NIb at the amino terminus of the polyprotein or in between P1 and HC-Pro were viable in tobacco plants.


Reevaluation of the sand rat as a model for diabetes mellitus.
The North African sand rat, Psammomys obesus, has been claimed to be a valuable model for investigating diabetes. It has been reported that, in the laboratory with free access to food, sand rats overeat, become obese, and develop hyperglycemia and ketoacidosis. We have observed that, although sand rats from our colony become obese and hyperinsulinemic, hyperglycemia is rare and ketoacidosis is virtually nonexistent. Because age, obesity, and dietary sucrose have been reported to be risk factors for diabetes, we have attempted to cause diabetes in a large group of sand rats by inducing obesity through sucrose feeding over an extended period of time. Animals fed sucrose-rich diets were compared to controls fed normal carbohydrate (starch) diets. No group of animals studied over 18 mo developed hyperglycemia and virtually all groups maintained small but significant glucose-induced acute insulin responses. Hyperinsulinemia and insulin resistance accompanied obesity, but there were no differences between dietary groups. Lean sand rats fed sucrose had higher circulating levels of triglyceride (TG) and accelerated TG secretion rates (TGSR) compared to lean controls. After a longer period of observation, all sand rats had elevated plasma TG levels, however, TGSR were normal, indicating that TG removal may have become defective. Although this animal is an excellent model in which to examine spontaneous obesity, Psammomys obesus is a poor model for the study of human diabetes mellitus.


Electroretinogram in sucrose-fed diabetic rats treated with an aldose reductase inhibitor or an anticoagulant.
To investigate the role of increased polyol pathway activity and hemodynamic deficits in the pathogenesis of diabetic retinopathy in non-insulin-dependent diabetes mellitus (NIDDM), Otsuka Long-Evans Tokushima fatty (OLETF) rats, an animal model of human NIDDM, were given water with or without 30% sucrose and some of them were fed laboratory chow containing 0.03% cilostazol, an anticoagulant, or 0.05% [5-(3-thienyl)tetrazol-1-yl] acetic acid monohydrate (TAT), an aldose reductase inhibitor, for 8 wk. Long-Evans Tokushima Otsuka (LETO) rats were used as nondiabetic controls. The peak latencies of oscillatory potentials of the electroretinogram in sucrose-fed OLETF rats were significantly prolonged compared with those in OLETF rats without sucrose feeding and LETO rats. There was a marked increase in platelet aggregability and a significant decrease in erythrocyte 2,3-diphosphoglycerate in sucrose-fed OLETF rats. Cilostazol significantly improved these parameters without changes in retinal levels of sorbitol and fructose. TAT, however, ameliorated all of these parameters. These findings confirm that the sucrose-fed OLETF rat is a useful animal model of retinopathy in human NIDDM and suggest that cilostazol improved diabetic retinopathy by modifying vascular factors, not by altering polyol pathway activity.


Funding child rearing: child allowance and parental leave.
This article proposes two financing plans to address what the author identifies as the two primary concerns in the child care field: (1) a child allowance for poor and near-poor households to address the child care problems of low-income families, and (2) a program of voluntary parental leave, available to all parents at child birth or adoption, to ensure the adequacy of infant care. The child allowance plan would cover the first three children in families up to 175% of the poverty level (more than 22 million children) at an annual cost of $45 billion. The author suggests that the allowance could be financed by redirecting funds from existing income support (for example, Aid to Families with Dependent Children), tax credit, and tax deduction programs. Financing the parental leave program would require new revenues, generated by an employee-paid increase in payroll tax totaling 3.5%. Each employee's contributions would create a parental leave account (PLA). Families could use the funds in these accounts to cover the cost of a one-year leave from work after the birth or adoption of a child. If families did not have enough dollars in their accounts to cover the cost of the leave, the federal government would extend a low-interest loan to them, which they would have to pay back. The amount individuals receive through Social Security would be adjusted upward or downward according to the balances in their parental leave accounts at retirement. The author suggests that both proposals would help parents balance work and family obligations and protect parental freedom of choice over the care and upbringing of their children.


Altered cortical activation associated with mirror overflow driven by non-dominant hand movement in attention-deficit/hyperactivity disorder.
Mirror overflow is involuntary movement that accompanies unilateral voluntary movement on the opposite side of the body, and is commonly seen in Attention-Deficit/Hyperactivity Disorder (ADHD). Children with ADHD show asymmetry in mirror overflow between dominant and non-dominant hand, yet there are competing mechanistic accounts of why this occurs. Using EEG during a sequential, unimanual finger-tapping task, we found that children with ADHD exhibited significantly more mirror overflow than typically developing (TD) controls, especially during the tapping of the non-dominant hand. Furthermore, source-level EEG oscillation analysis revealed that children with ADHD showed decreased alpha (8-12 Hz) event-related desynchronization (ERD) compared with controls in both hemispheres, but only during tapping of the non-dominant hand. Moreover, only the ERD ipsilateral to the mirror overflow during non-dominant hand movement correlated with both magnitude of overflow movements and higher ADHD symptom severity (Conners ADHD Hyperactivity/Impulsiveness scale) in children with ADHD. TD controls did not show these relationships. Our findings suggest that EEG differences in finger-tapping in ADHD are related primarily to voluntary movement in the non-dominant hand. Our results are also consistent with the Ipsilateral Corticospinal Tract (CST) Hypothesis, which posits that the atypical persistence of mirror overflow in ADHD may originate in the sensorimotor areas ipsilateral to mirror overflow and be transmitted via non-decussating CST fibers.


Steers grazing blue grama rangeland throughout the growing season. II. Site and extent of digestion and microbial protein synthesis.
Effects of advancing forage maturity and drought-induced summer dormancy on site and extent of digestion and microbial protein synthesis in beef steers grazing native blue grama rangeland were evaluated in four sampling periods. Five steers (avg initial wt 227 kg) fitted with ruminal, duodenal and ileal cannulae and three steers cannulated at the esophagus freely grazed a 12-ha study pasture. Sampling periods lasted 11 d and started June 2, which was during the early growing season (EGS); June 22, during early summer dormancy (ESD); July 21, during late summer dormancy (LSD); and August 25, 1985, during the late growing season (LGS). Dietary N content was lower (P less than .05) in ESD and LSD than in EGS and LGS. Neutral detergent fiber (NDF) content was lower (P less than .05) in EGS than in other sampling periods. Ruminal organic matter (OM) digestion was lower (P less than .05) in ESD than in EGS, probably because of increased dietary NDF and lower N content. Ruminal OM digestion was greater (P less than .05) in LSD and LGS than in ESD because of increased fiber digestion. Neutral detergent fiber and acid detergent fiber (ADF) digestion occurring in the rumen was greater (P less than .05) in LSD and LGS than in EGS and ESD. Organic matter digestion in the small intestine and OM, NDF and ADF digestion in the hindgut were similar for all sampling periods. Over 90% of the fiber digestion occurred ruminally.(ABSTRACT TRUNCATED AT 250 WORDS)


Stress reactivity and social cognition in pure and co-occurring early childhood relational bullying and victimization.
This study tested the independent effects and interactions of sympathetic nervous system reactivity and hostile attribution biases (HAB) in predicting change in pure and co-occurring relational bullying and victimization experiences over one year. Co-occurring and pure relational bullying and victimization experiences were measured using a dimensional bifactor model, aiming to address methodological limitations of categorical approaches, using data from 300 preschoolers (Mage = 44.70 months, SD = 4.38). Factor scores were then saved and used in nested path analyses with a subset of participants (n = 81) to test main study hypotheses regarding effects of HAB and skin conductance level reactivity (SCL-R). Bifactor models provided good fit to the data at two independent time points. HAB and SCL-R interacted to predict increases in co-occurring relational bullying/victimization with evidence for over- and underarousal pathways.


Alternative transcription start sites of the enolase-encoding gene enoA are stringently used in glycolytic/gluconeogenic conditions in Aspergillus oryzae.
Gene expression using alternative transcription start sites (TSSs) is an important transcriptional regulatory mechanism for environmental responses in eukaryotes. Here, we identify two alternative TSSs in the enolase-encoding gene (enoA) in Aspergillus oryzae, an industrially important filamentous fungus. TSS use in enoA is strictly dependent on the difference in glycolytic and gluconeogenic carbon sources. Transcription from the upstream TSS (uTSS) or downstream TSS (dTSS) predominantly occurs under gluconeogenic or glycolytic conditions, respectively. In addition to enoA, most glycolytic genes involved in reversible reactions possess alternative TSSs. The fbaA gene, which encodes fructose-bisphosphate aldolase, also shows stringent alternative TSS selection, similar to enoA. Alignment of promoter sequences of enolase-encoding genes in Aspergillus predicted two conserved regions that contain a putative cis-element required for enoA transcription from each TSS. However, uTSS-mediated transcription of the acuN gene, an enoA ortholog in Aspergillus nidulans, is not strictly dependent on carbon source, unlike enoA. Furthermore, enoA transcript levels in glycolytic conditions are higher than in gluconeogenic conditions. Conversely, acuN is more highly transcribed in gluconeogenic conditions. This suggests that the stringent usage of alternative TSSs and higher transcription in glycolytic conditions in enoA may reflect that the A. oryzae evolutionary genetic background was domesticated by exclusive growth in starch-rich environments. These findings provide novel insights into the complexity and diversity of transcriptional regulation of glycolytic/gluconeogenic genes among Aspergilli.


α-1,3-N-acetylgalactose aminotransferase gene 539G>C mutation leads to the A2B isoform.
In this study, the phenotypic identification and molecular mechanism of one case of an A2B subtype pedigree was investigated. ABO blood groupings were identified by serological methods and sequence amplification was performed by polymerase chain reaction (PCR) using TA cloning and DNA sequencing analysis to identify the pedigree and the ABO gene haploid of the proband. There were both A and B antigens on the proband's red blood cells, and anti-A1 antibodies were found in the serum. Direct sequencing of the 6th and 7th exons of the ABO gene showed the A208/B101 genotype, and haploid determination revealed the A208 and B101 alleles. Compared with the A102 allele sequence, the A208 allele was mutated at the 539 G>C site. Pedigree analysis showed that the ABO blood phenotypes of the proband's father, mother, husband, and daughter were A2, B, AB, and A2B, respectively, and their genotypes were A208/O02, B101/B101, A102/B101, and A208/B101, respectively. The father of the proband had anti-A1 antibodies and the A208 allele of the proband was inherited from her father, which can be passed on to her daughter. The α-1, 3-N-acetylgalactose aminotransferase gene 539G>C mutation resulted in A2B phenotype generation, and individual serum contained the anti-A1 antibody.


Cost-effectiveness analysis of spinal cord stimulation in treatment of failed back surgery syndrome.
This article presents an analysis of the medical costs of spinal cord stimulation (SCS) therapy in the treatment of patients with failed back surgery syndrome (FBSS). We compared the medical costs of SCS therapy with an alternative regimen of surgeries and other interventions. Externally powered (external) and fully internalized (internal) SCS systems were considered separately. Clinical management models of each of the therapy alternatives were derived from the clinical literature, retrospective data sets, expert opinion, and published diagnostic and therapy protocols. No value was placed on pain relief or improvements in the quality of life that successful SCS therapy can generate. We found that by reducing the demand for medical care by FBSS patients, SCS therapy can lower medical costs. On average, given current screening and efficacy rates, SCS therapy pays for itself within 5.5 years. For those patients for whom SCS therapy is clinically efficacious, the therapy pays for itself within 2.1 years.


Effect of glucagon on hepatic phenylalanine hydroxylase in vivo.
Moderate doses of glucagon (20 micrograms/kg I.V.) are sufficient to stimulate rat hepatic phenylalanine hydroxylase in vivo. In addition, the stimulation of the tetrahydrobiopterin-dependent phenylalanine hydroxylase activity in livers of animals fed on a high-protein diet has been correlated with an elevated phosphate content. The tetrahydrobiopterin-dependent hydroxylase activity in these animals can be further elevated by glucagon-stimulated phosphorylation. These results indicate that physiological changes in glucagon concentration modulate rat liver phenylalanine hydroxylase activity in vivo. The current understanding of the role of phosphorylation in regulating human phenylalanine hydroxylase is also considered.


Inhibitory activities of lansoprazole against respiration in Helicobacter pylori.
Lansoprazole and its derivative AG-1789 dose-dependently inhibited cellular respiration by an endogenous substrate and decreased the ATP level in Helicobacter pylori cells. The inhibitory action of lansoprazole and AG-1789 against respiration was specific to substrates such as pyruvate and alpha-ketoglutarate and similar to the inhibitory action of rotenone, which is an inhibitor for the mitochondrial respiratory chain. Growth inhibition by lansoprazole and AG-1789 as well as by rotenone was augmented at high oxygen concentrations under atmospheric conditions. Since the 50% inhibitory concentrations of these compounds for the respiration were close to their MICs for H. pylori growth, the growth inhibition might be due to respiratory inhibition by these compounds.


Clinical assessment of sexual trauma: interviewing adult survivors of childhood abuse.
Clinicians often feel reticent about exploring sexual trauma and its aftermath too soon. The author contends that, properly implemented, early exploration can actually foster the treatment of patients who have experienced sexual trauma. Appropriate clinical assessment can promote a therapeutic relationship and help both clinician and patient develop a comprehensive view of how the past trauma continues to influence the patient's current life. The author presents a detailed assessment outline and suggests that the findings from such an assessment can serve as an effective guide for further therapeutic work.


Primary treatment of ruptured blood blister-like aneurysms with stent-assisted coil embolization: report of two cases.
Blood blister-like aneurysms (BBAs) are among the most hazardous cerebrovascular aneurysms to treat; microsurgical treatment of these small, wide-necked, and exceptionally fragile aneurysms place patients at significant risk of morbidity or mortality. We report two cases of ruptured BBAs attempted to be treated for the first time with stent-assisted coil embolization solely and review the current literature on treatment options. Our patients underwent stent-assisted coil embolization of the aneurysms in the acute stage of subarachnoid hemorrhage (SAH). One patient was successfully treated without procedure-related complications. The other patient died after surgical internal carotid artery (ICA) occlusion, carried out after intraoperative rerupture of the aneurysm during the endovascular treatment. In the successful case, 8-month and 19-month follow-up angiograms demonstrated incomplete (>90%) occlusion with residual filling of the aneurysm neck, which did not need additional coil embolization. Even though stent-assisted coil embolization of ruptured BBAs in the acute stage appears to be a technically feasible treatment option, the present stent-related endovascular technology has potentially hazardous drawbacks.


Hemolytic complement activity in aqueous humor.
Hemolytic C1, C4, C3, and C5 and IgG levels were measured in aqueous humor obtained at surgery from human eyes without inflammation or previous surgery (group 1), eyes with previous surgery (group 2), and eyes with anterior uveitis (group 3). Serum hemolytic complement and IgG levels were also compared with aqueous humor levels. Aqueous humor from eyes in group 1 contained functional C1, C4, C3, and C5, but there was relatively little complement in normal aqueous humor when compared with serum. The mean values of all complement components and IgG in aqueous humor increased from groups 1 to 3; aqueous-serum ratios for each complement component and IgG also increased from groups 1 to 3. Eyes in group 3 had the highest complement and IgG levels and in some cases these values approached those found in serum.


Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) Inhibitors, Reality or Dream in Managing Patients with Cardiovascular Disease.
Statins have been a major keystone in the management of patients with atherosclerotic cardiovascular disease. The benefits of inhibiting HMG CoA reductase, via statins, were translated into reduction in LDL-c with proportionate decrease in cardiovascular events in response to the magnitude of LDL-c reduction. Despite major advances in pharmacological treatments, including the use of high-dose statins, there are urgent need to further reduce future cardiovascular risk. This is in particularly important since 1 out of 5 high-risk atherosclerotic patients who achieve low LDL-c return with a second cardiovascular event within five years. Although this residual risk post-statin is largely heterogeneous, lowering LDL-c beyond 'normal' or guidelines-recommended level using novel therapies has resulted in further reduction in cardiovascular events. The current review will discuss the use of PCSK9 inhibitors in patients with atherosclerotic disease. PCSK9 inhibitors are a new class of lipid-lowering drugs that are either fully human monoclonal antibodies (evolocumab and alirocumab) or humanised monoclonal antibodies (bococizumab) that effectively reduce LDL-c to unprecedented level. By blocking circulating PCSK9, these drugs would preserve LDL receptors and prevent them from cellular degradation. This process promotes recycling of LDL receptors back to hepatocytes surface, leading into further reduction of LDL-c. Combining PCSK9 inhibitors with statin have led into lower LDL-c, reduction in plaque volume and more importantly reduction in future cardiovascular events. These drugs are very promising, nonetheless, the unselective approach of applying these monoclonal antibodies may not prove to be cost-effective and potentially exposing some patients to unnecessary side effects.


Knowledge of contraceptive effectiveness and method use among women in Hanoi, Vietnam.
To evaluate the association between contraceptive knowledge and type of method used. We analyzed data from a cross-sectional study of sexually active women in Hanoi, Vietnam, not desiring pregnancy. We used linear and logistic regression to evaluate contraceptive knowledge of users of the intrauterine device (IUD), combination oral contraception (COC) and male condoms. We measured contraceptive knowledge with seven questions on relative effectiveness of methods, reversibility, covert use, contraindications and side effects. Respondents used IUD (n = 128), COC (n = 126) or condoms (n = 167). Summary knowledge scores did not differ by current type of method used. Only one knowledge domain, contraceptive effectiveness, varied by method. Compared to condom users, IUD users had higher odds of correctly identifying the IUD as more effective than COC, condoms and withdrawal (adjusted odds ratio [aOR], 4.8; 95% confidence interval [CI], 2.7-8.3). Higher proportions of condom users (49.7%) mistakenly identified condoms as the most effective of listed methods compared to IUD (20.3%) and COC users (23.0%). On the other hand, IUD and COC users had lower odds (aOR, 0.5; 95% CI, 0.2-1.0 and aOR, 0.3; 95% CI, 0.1-0.6, respectively) of identifying consistent condom use as better for pregnancy prevention than other practices (e.g., withdrawal and postcoital douching). IUD users more often recognized that the IUD is highly effective while condom users appeared to overestimate condom effectiveness. Contraceptive counseling should ensure that women understand the relative effectiveness of methods. We found no evidence that other types of contraceptive knowledge differed by type of method used. Knowledge of contraceptive effectiveness was the sole difference detected in contraceptive knowledge between women in Hanoi, Vietnam, using the IUD, COC or male condoms.


Assay of protein kinase C with an N-bromosuccinimide-cleavage fragment of histone H1.
N-Bromosuccinimide cleavage of a lysine-rich histone fraction (histone III-S) yields a peptide substrate, purified by reverse-phase h.p.l.c., for the Ca2+ + phospholipid-dependent protein kinase (protein kinase C). This substrate displays no reactivity with the cyclic AMP-dependent protein kinase, and may prove useful for the detection of protein kinase C activity in crude tissue extracts.


Metergoline in the management of normoprolactinemic secondary amenorrhea. A multicentric controlled trial.
In a multicentric study, the effect of the antiserotoninergic agent metergoline was evaluated in the management of patients with idiopathic normoprolactinemic secondary amenorrhea (NSA). The awareness that psychological factors might lead to a spontaneous reappearance of menses was also taken into account, and all the patients, after physical, gynecological and laboratory examinations, and the performance of the progesterone withdrawal bleeding test (100 mg i.m.) and the clomiphene citrate test (100 mg p.o./day for 5 days), were treated for 60 days with placebo; only patients showing no menses during placebo administration were later treated with metergoline. 108 patients entered the trial: of these, 48 experienced menses on admission or during placebo administration, and were withdrawn. Of the 60 patients not responding to placebo, 50 were treated for 90 days with metergoline (4 mg t.i.d.), and 23 had menses, ovulatory in 68.4% of cases. A new placebo treatment was accompanied, in the majority of cases, by recurrence of amenorrhea. These results indicate that many patients with NSA may experience a spontaneous disappearance of the disease: in cases more seriously affected metergoline might be a useful therapeutic agent.


Molecular signatures of host specificity linked to habitat specialization in Exaiptasia sea anemones.
Rising ocean temperatures associated with global climate change induce breakdown of the symbiosis between coelenterates and photosynthetic microalgae of the genus Symbiodinium. Association with more thermotolerant partners could contribute to resilience, but the genetic mechanisms controlling specificity of hosts for particular Symbiodinium types are poorly known. Here, we characterize wild populations of a sea anemone laboratory model system for anthozoan symbiosis, from contrasting environments in Caribbean Panama. Patterns of anemone abundance and symbiont diversity were consistent with specialization of holobionts for particular habitats, with Exaiptasia pallida/S. minutum (ITS2 type B1) abundant on vertical substrate in thermally stable, shaded environments but E. brasiliensis/Symbiodinium sp. (ITS2 clade A) more common in shallow areas subject to high temperature and irradiance. Population genomic sequencing revealed a novel E. pallida population from the Bocas del Toro Archipelago that only harbors S. minutum. Loci most strongly associated with divergence of the Bocas-specific population were enriched in genes with putative roles in cnidarian symbiosis, including activators of the complement pathway of the innate immune system, thrombospondin-type-1 repeat domain proteins, and coordinators of endocytic recycling. Our findings underscore the importance of unmasking cryptic diversity in natural populations and the role of long-term evolutionary history in mediating interactions with Symbiodinium.


Phenytoin-induced asterixis--uncommon or under-diagnozed?
Asterixis is a recognized but uncommon clinical sign of phenytoin toxicity. A case is presented in which asterixis and acute cerebellar dysfunction occurred when the phenytoin levels reached the toxic range. It disappeared when the drug levels normalized. Asterixis is now classified as a form of negative myoclonus, which is characterized by irregular myoclonic lapses of posture. The neurochemistry and physiology of phenytoin causing asterixis has yet to be elucidated.


Positional therapy in sleep apnoea - one fits all? What determines success in positional therapy in sleep apnoea syndrome.
Positional therapy is a simple means of therapy in sleep apnoea syndrome, but due to controversial or lacking evidence, it is not widely accepted as appropriate treatment. In this study, we analysed data to positional therapy with regard to successful reduction of AHI and predictors of success. All consecutive patients undergoing polysomnography between 2007 and 2011 were analysed. We used a strict definition of positional sleep apnoea syndrome (supine-exclusive sleep apnoea syndrome) and of therapy used. Patients underwent polysomnography initially and during follow-up. 1275 patients were evaluated, 112 of which had supine-exclusive sleep apnoea syndrome (AHI 5-66/h, median 13/h), 105 received positional therapy. With this treatment alone 75% (70/105) reached an AHI <5/h, in the follow-up 1 year later 37% (37/105) of these still had AHI<5/h, 46% (43/105) yielded an AHI between 5 and 10/h. Nine patient switched to APAP due to deterioration, 3 wanted to try APAP due to comfort reasons. At the last follow-up, 32% patients (34/105) were still on positional therapy with AHI <5/h. BMI was a predictor for successful reduction of AHI, but success was independent of sex, the presence of obstructive versus central sleep apnoea, severity of sleep apnoea syndrome or co-morbidities. Positional therapy may be a promising therapy option for patients with positional sleep apnoea. With appropriate adherence it yields a reasonable success rate in the clinical routine.


In vitro metabolism of amiodarone by rabbit and rat liver and small intestine.
The present studies were designed to investigate whether amiodarone (Am) is metabolized in the major organs and tissues of the rat and rabbit. Incubations using Am and tissue homogenates (600 g supernatant) of rabbit and rat lung, liver, kidney, and gut revealed formation of desethylamiodarone (DEA) by the liver and gut. Subsequent experiments using the post-mitochondrial, cytosolic, and microsomal fractions of these tissues indicated that metabolism of Am was greatest in the microsomal fractions. In both species, greater DEA formation was detected for microsomes of hepatic origin. The hepatic microsomal mediated production of DEA was altered by protein concentration in both the rabbit and rat preparations with protein concentrations of 5 mg providing the greatest DEA production. DEA formation by gut microsomes was greatest at 3 mg of protein for the rabbit but exhibited no significant change from 1 mg to 10 mg of protein for the rat. In vitro metabolism of Am by rabbit and rat hepatic microsomal preparations was significantly reduced by 1 mM piperonyl butoxide, SKF 525-A, n-octylamine, and carbon monoxide. Effects of these inhibitors on rabbit and rat gut microsomal incubations were inconclusive. HPLC analysis of incubation samples revealed a species difference in the metabolism of Am as demonstrated by the detection of three metabolites in addition to DEA. The unidentified metabolites (I, II, III) were detected in rabbit hepatic microsomal incubations. Metabolite II was also detected in incubations using rabbit duodenal tissue microsomes. No metabolites other than DEA were found in incubations using rat tissues.(ABSTRACT TRUNCATED AT 250 WORDS)


Construction of periventricular white matter hyperintensity maps by spatial normalization of the lateral ventricles.
Subcortical and periventricular white matter hyperintensities (WMHs) may have different associations with cognition and pathophysiology. The aim of the present study is to develop an automated method for construction of periventricular WMH maps that enables the analysis of between-group differences in WMH location and characteristics in the periventricular region without the requirement of prior boundary definition. To avoid influence of WMHs on spatial normalization, a reference image of the lateral ventricles was constructed based on images of 24 subjects. Construction was not biased to a single subject. WMHs were segmented by k-nearest neighbor-based classification of magnetic resonance inversion recovery and fluid attenuated inversion recovery images. Cerebrospinal fluid segmentations of individual subjects were nonrigidly mapped to the reference image of the lateral ventricles. The subject's WMHs were transformed to the reference space accordingly. Spatial normalization accuracy was validated using measures of overlap and of displacement relative to the boundary of the lateral ventricles. After spatial normalization, the boundaries of the lateral ventricles closely matched the reference image and in an area of approximately 1 cm around the lateral ventricles the relative displacement was less than 1 mm. To illustrate the method, it was applied to 61 patients with Type 2 diabetes and 26 control subjects, whereupon periventricular WMH maps were constructed and compared. The proposed method is particularly suited to analyze WMH distribution differences at the level of the lateral ventricles between large groups of patients.


Roles of DNA repair and reductase activity in the cytotoxicity of the hypoxia-activated dinitrobenzamide mustard PR-104A.
PR-104 is a dinitrobenzamide mustard currently in clinical trial as a hypoxia-activated prodrug. Its major metabolite, PR-104A, is metabolized to the corresponding hydroxylamine (PR-104H) and amine (PR-104M), resulting in activation of the nitrogen mustard moiety. We characterize DNA damage responsible for cytotoxicity of PR-104A by comparing sensitivity of repair-defective hamster Chinese hamster ovary cell lines with their repair-competent counterparts. PR-104H showed a repair profile similar to the reference DNA cross-linking agents chlorambucil and mitomycin C, with marked hypersensitivity of XPF(-/-), ERCC1(-/-), and Rad51D(-/-) cells but not of XPD(-/-) or DNA-PK(CS)(-/-) cells. This pattern confirmed the expected dependence on the ERCC1-XPF endonuclease, implicated in unhooking DNA interstrand cross-links at blocked replication forks, and homologous recombination repair (HRR) in restarting collapsed forks. However, even under anoxia, the hypersensitivity of XPF(-/-), ERCC1(-/-), and Rad51D(-/-) cells to PR-104A itself was lower than for chlorambucil. To test whether this reflects inefficient PR-104A reduction, a soluble form of human NADPH:cytochrome P450 oxidoreductase was stably expressed in Rad51D(-/-) cells and their HRR-restored counterpart. This expression increased hypoxic metabolism of PR-104A to PR-104H and PR-104M as well as hypoxia-selective cytotoxicity of PR-104A and its dependence on HRR. We conclude that PR-104A cytotoxicity is primarily due to DNA interstrand cross-linking by its reduced metabolites, although under conditions of inefficient PR-104A reduction (low reductase expression or aerobic cells), a second mechanism contributes to cell killing. This study shows that hypoxia, reductase activity, and DNA interstrand cross-link repair proficiency are key variables that interact to determine PR-104A sensitivity.


Patterns of HER2 Gene Amplification and Response to Anti-HER2 Therapies.
A chromosomal region that includes the gene encoding HER2, a receptor tyrosine kinase (RTK), is amplified in 20% of breast cancers. Although these tumors tend to respond to drugs directed against HER2, they frequently become resistant and resume their malignant progression. Gene amplification in double minutes (DMs), which are extrachromosomal entities whose number can be dynamically regulated, has been suggested to facilitate the acquisition of resistance to therapies targeting RTKs. Here we show that ~30% of HER2-positive tumors show amplification in DMs. However, these tumors respond to trastuzumab in a similar fashion than those with amplification of the HER2 gene within chromosomes. Furthermore, in different models of resistance to anti-HER2 therapies, the number of DMs containing HER2 is maintained, even when the acquisition of resistance is concomitant with loss of HER2 protein expression. Thus, both clinical and preclinical data show that, despite expectations, loss of HER2 protein expression due to loss of DMs containing HER2 is not a likely mechanism of resistance to anti-HER2 therapies.


Extensive elimination of acinar cells during normal postnatal pancreas growth.
While programmed cell death plays important roles during morphogenetic stages of development, post-differentiation organ growth is considered an efficient process whereby cell proliferation increases cell number. Here we demonstrate that early postnatal growth of the pancreas unexpectedly involves massive acinar cell elimination. Measurements of cell proliferation and death in the human pancreas in comparison to the actual increase in cell number predict daily elimination of 0.7% of cells, offsetting 88% of cell formation over the first year of life. Using mouse models, we show that death is associated with mitosis, through a failure of dividing cells to generate two viable daughters. In p53-deficient mice, acinar cell death and proliferation are reduced, while organ size is normal, suggesting that p53-dependent developmental apoptosis triggers compensatory proliferation. We propose that excess cell turnover during growth of the pancreas, and presumably other organs, facilitates robustness to perturbations and supports maintenance of tissue architecture.


Analysis of the role of TFIIE in basal transcription and TFIIH-mediated carboxy-terminal domain phosphorylation through structure-function studies of TFIIE-alpha.
The general transcription factor TFIIE recruits TFIIH at a late stage of transcription initiation complex formation and markedly stimulates TFIIH-dependent phosphorylation of the carboxy-terminal domain (CTD) of RNA polymerase II. To study this function of TFIIE in more detail, systematic deletion mutations were introduced into the large subunit of TFIIE (TFIIE-alpha) and were analyzed with regard to their effects on TFIIH-dependent CTD phosphorylation, TFIIE-dependent basal and enhancer-dependent transcription, and interactions of TFIIE-alpha with both TFIIE-beta and TFIIH. The amino (N)-terminal half of TFIIE-alpha, which possesses several putative structural motifs, was sufficient for the phosphorylation and transcription activities and for TFIIE-beta interactions, whereas a site effecting both strong interactions with TFIIH and large stimulatory effects on transcription and CTD phosphorylation was localized to an acidic region near the carboxy (C) terminus. The fact that these activities appear to be tightly linked supports the idea that TFIIE interacts physically and functionally with TFIIH and that CTD phosphorylation is essential for transcription under normal conditions. The present results suggest that TFIIE, via its effect on TFIIH, may act as a checkpoint for formation of a preinitiation complex.


RETROSPECTIVE STUDY OF CAUSES OF ANIMAL MORTALITY IN LJUBLJANA ZOO 2005-2015.
Zoos harbor large collections of diverse species, aiding in both conservation and education, as well as research in multiple scientific fields. However, the most common causes of death in zoo animals around the world remain unclear because few extensive reports or reviews are published on this topic. This information could greatly improve preventive veterinary medicine in zoologic gardens. This study provides a retrospective overview of the causes of death of animals from the Ljubljana Zoo in the years 2005-2015. During this period, a total of 353 animals were submitted for necropsy, of which 244 were mammals, 85 were birds, and 25 were reptiles. The causes of deaths were divided into infectious diseases (38%), dysfunctions of individual organs (20%), traumas (13%), parasitosis (7%), reproductive disorders (6%), metabolic disorders (3%), neoplastic disease (4%), and intoxications (4%). In some cases, the cause of death was unable to be determined (7%), most commonly because of autolysis of the body. The results of this retrospective study bring a general overview of the epizootiologic situation in the Ljubljana Zoo over an 11-yr period and valuable information to other zoos to optimize preventative plans and diagnostics.


Translation and cross-cultural adaptation of a Nepali version of the Dutch Participation and Activity Inventory for Children and Youth (PAI - CY) with visual impairment.
Visual impairment is an important cause of disability in children. There is a lack of information on rehabilitation needs and low vision support services for children with visual impairment in Nepal. This is a pilot study designed to translate, culturally adapt and pre-test the Dutch version of the Participation and Activity Inventory for Children and Youth (PAI-CY) with visual impairment aged 7-17 years to develop a Nepali version. Questionnaires (PAI-CY versions for 7-12 and 13-17 years) were translated using standardized methods and were culturally adapted by a panel of experts. They were pretested to evaluate comprehensibility and relevance among six children with visual impairment and blindness. Finally, participants completed a questionnaire evaluation form. The translation and cultural adaptation process resulted in the adaptation of nine items to make them suitable for Nepali culture. Most children had comprehensibility problems with some specific items because of vocabulary, sentence structure and the composition of items. Most of the children were satisfied with the questionnaires. The study resulted in the development of a Nepali version of the PAI-CY. We worked with a small group of content experts and a small but representative sample of children which allowed us to use rigorous translation procedures to address language and cultural differences. A population based study has been planned to investigate the psychometric properties of these questionnaires.


Association of axon guidance factor semaphorin 3A with poor outcome in pancreatic cancer.
Neural alterations and aberrantly expressed nerve-specific factors promoting tumor progression are known to contribute to pancreatic cancer's extremely poor prognosis. Despite hints that axon guidance factor semaphorin 3A (SEMA3A) may function as a tumor inhibitor, its clinical importance and therapeutic potential have not yet been explored. The present study investigated the role of SEMA3A and its receptors-plexins A1-A4 (PLXNA1-A4) and neuropilin-1 (NRP1)-in pancreatic cancer. QRT-PCR and immunohistochemical analyses revealed overexpression of SEMA3A, NRP1 and PLXNA1 in metaplastic ducts, malignant cells and nerves of cancerous specimens, and showed that elevated levels of corresponding mRNA (6.8-fold, 2.0-fold and 1.5-fold, respectively) clearly correlated with negative clinicopathological manifestations such as shorter survival (SEMA3A and PLXNA1) and a lesser degree of tumor differentiation (NRP1) in Stages I-III patients. High SEMA3A expression in pancreata of Stage IV M1 patients and in peritoneal metastases, and consequent functional studies indicated that poor clinical outcome might be related to the ability of SEMA3A to promote dissemination and invasiveness of pancreatic cancer cells through activation of multiple pathways involving Rac1, GSK3b or p42/p44 MAPK, but not E- to N-cadherin switch, MMP-9 or VEGF induction. Thus, this study is the first to quantify expression of the SEMA3A system in human malignancy and to show that overexpression of SEMA3A by nerves and transformed cells leads to a SEMA3A-rich environment which may favor malignant activities of tumor cells. Furthermore, negative clinicopathological correlations suggest that SEMA3A might represent a novel intervention target but not a treatment option for pancreatic cancer patients.


Infectious Entry of Merkel Cell Polyomavirus.
Merkel cell polyomavirus (MCPyV) is a small, nonenveloped tumor virus associated with an aggressive form of skin cancer, Merkel cell carcinoma (MCC). MCPyV infections are highly prevalent in the human population, with MCPyV virions being continuously shed from human skin. However, the precise host cell tropism(s) of MCPyV remains unclear: MCPyV is able to replicate within a subset of dermal fibroblasts, but MCPyV DNA has also been detected in a variety of other tissues. However, MCPyV appears different from other polyomaviruses, as it requires sulfated polysaccharides, such as heparan sulfates and/or chondroitin sulfates, for initial attachment. Like other polyomaviruses, MCPyV engages sialic acid as a (co)receptor. To explore the infectious entry process of MCPyV, we analyzed the cell biological determinants of MCPyV entry into A549 cells, a highly transducible lung carcinoma cell line, in comparison to well-studied simian virus 40 and a number of other viruses. Our results indicate that MCPyV enters cells via caveolar/lipid raft-mediated endocytosis but not macropinocytosis, clathrin-mediated endocytosis, or glycosphingolipid-enriched carriers. The viruses were internalized in small endocytic pits that led the virus to endosomes and from there to the endoplasmic reticulum (ER). Similar to other polyomaviruses, trafficking required microtubular transport, acidification of endosomes, and a functional redox environment. To our surprise, the virus was found to acquire a membrane envelope within endosomes, a phenomenon not reported for other viruses. Only minor amounts of viruses reached the ER, while the majority was retained in endosomal compartments, suggesting that endosome-to-ER trafficking is a bottleneck during infectious entry.IMPORTANCE MCPyV is the first polyomavirus directly implicated in the development of an aggressive human cancer, Merkel cell carcinoma (MCC). Although MCPyV is constantly shed from healthy skin, the MCC incidence increases among aging and immunocompromised individuals. To date, the events connecting initial MCPyV infection and subsequent transformation still remain elusive. MCPyV differs from other known polyomaviruses concerning its cell tropism, entry receptor requirements, and infection kinetics. In this study, we examined the cellular requirements for endocytic entry as well as the subcellular localization of incoming virus particles. A thorough understanding of the determinants of the infectious entry pathway and the specific biological niche will benefit prevention of virus-derived cancers such as MCC.


AtNPF2.5 Modulates Chloride (Cl-) Efflux from Roots of Arabidopsis thaliana.
The accumulation of high concentrations of chloride (Cl-) in leaves can adversely affect plant growth. When comparing different varieties of the same Cl- sensitive plant species those that exclude relatively more Cl- from their shoots tend to perform better under saline conditions; however, the molecular mechanisms involved in maintaining low shoot Cl- remain largely undefined. Recently, it was shown that the NRT1/PTR Family 2.4 protein (NPF2.4) loads Cl- into the root xylem, which affects the accumulation of Cl- in Arabidopsis shoots. Here we characterize NPF2.5, which is the closest homolog to NPF2.4 sharing 83.2% identity at the amino acid level. NPF2.5 is predominantly expressed in root cortical cells and its transcription is induced by salt. Functional characterisation of NPF2.5 via its heterologous expression in yeast (Saccharomyces cerevisiae) and Xenopus laevis oocytes indicated that NPF2.5 is likely to encode a Cl- permeable transporter. Arabidopsis npf2.5 T-DNA knockout mutant plants exhibited a significantly lower Cl- efflux from roots, and a greater Cl- accumulation in shoots compared to salt-treated Col-0 wild-type plants. At the same time, [Formula: see text] content in the shoot remained unaffected. Accumulation of Cl- in the shoot increased following (1) amiRNA-induced knockdown of NPF2.5 transcript abundance in the root, and (2) constitutive over-expression of NPF2.5. We suggest that both these findings are consistent with a role for NPF2.5 in modulating Cl- transport. Based on these results, we propose that NPF2.5 functions as a pathway for Cl- efflux from the root, contributing to exclusion of Cl- from the shoot of Arabidopsis.


VARP and Rab9 Are Dispensable for the Rab32/BLOC-3 Dependent Salmonella Killing.
Salmonella enterica serovar Typhi (S. Typhi) is the causative agent of typhoid fever, a disease that kills an estimated 200,000 people annually. Previously, we discovered an antimicrobial pathway dependent on Rab32 and BLOC-3 (BRAM) that is critical to kill S. Typhi in murine macrophages. The BLOC-3 complex is comprised of the two sub-units HPS1 and HPS4 and exhibits guanine-nucleotide exchange factor (GEF) activity to Rab32. In melanocytes, Rab9 has been shown to interact with HPS4 and RUTBC1, a Rab32 GTPase activating (GAP) protein, and regulate the Rab32-mediated melanosome biogenesis. Intriguingly, Rab9-deficient melanocytes exhibit hypopigmentation, a similar phenotype to Rab32 or BLOC-3 deficient melanocytes. Additionally, VPS9-ankyrin-repeat-protein (VARP) has been shown to regulate melanocytic enzyme trafficking into the melanosomes through interaction with Rab32. Although Rab32, Rab9 and VARP are a part of melanogenesis in melanocytes, whether Rab9 and VARP are required for the BRAM mediated killing in macrophages is currently unknown. Here we showed that HPS4 is recruited to the Salmonella-containing vacuoles (SCV) and over-expression of BLOC-3 significantly increased Rab32-positive bacteria vacuoles. We found that SCV acquire Rab9, however over-expressing Rab9 did not change HPS4 localization on bacteria vacuoles. Importantly, we used shRNA to knock-down Rab9 and VARP in macrophages and showed that these proteins are dispensable for Rab32 recruitment to the SCV. Furthermore, we assessed the survival of S. Typhimurium in macrophages deficient for Rab9 or VARP and demonstrated that these proteins are not essential for BRAM pathway-dependent killing.


Automated measurement of vocal fold vibratory asymmetry from high-speed videoendoscopy recordings.
In prior work, a manually derived measure of vocal fold vibratory phase asymmetry correlated to varying degrees with visual judgments made from laryngeal high-speed videoendoscopy (HSV) recordings. This investigation extended this work by establishing an automated HSV-based framework to quantify 3 categories of vocal fold vibratory asymmetry. HSV-based analysis provided for cycle-to-cycle estimates of left-right phase asymmetry, left-right amplitude asymmetry, and axis shift during glottal closure for 52 speakers with no vocal pathology producing comfortable and pressed phonation. An initial cross-validation of the automated left-right phase asymmetry measure was performed by correlating the measure with other objective and subjective assessments of phase asymmetry. Vocal fold vibratory asymmetry was exhibited to a similar extent in both comfortable and pressed phonations. The automated measure of left-right phase asymmetry strongly correlated with manually derived measures and moderately correlated with visual-perceptual ratings. Correlations with the visual-perceptual ratings remained relatively consistent as the automated measure was derived from kymograms taken at different glottal locations. An automated HSV-based framework for the quantification of vocal fold vibratory asymmetry was developed and initially validated. This framework serves as a platform for investigating relationships between vocal fold tissue motion and acoustic measures of voice function.


Neural activation state determines behavioral susceptibility to modified theta burst transcranial magnetic stimulation.
Transcranial magnetic stimulation (TMS) allows one to investigate the effects of temporary interference of neural processing in neurologically intact subjects. In a previous study [J. Silvanto et al. (2007) Eur. J. Neurosci., 25, 1874-1881] we found that online TMS perceptually facilitates the attributes encoded by the least active neural populations. The objective of the present experiment was to extend this work to determine whether such state-dependent effects can be observed when offline high-frequency TMS is applied to suppress neural activity. The activity levels of direction-selective neural populations in the V1/V2 region were modulated by asking subjects to passively view either leftward or rightward motion during offline TMS. In a subsequent motion direction-discrimination task, their ability to discriminate motion direction was dependent on the type of motion they had passively viewed during offline TMS: detection of the congruent direction (i.e. direction viewed during offline TMS) was unaffected, whereas detection of the incongruent direction (i.e. opposite direction to the one viewed during offline TMS) was impaired. As the activity level of neurons tuned to the incongruent direction was presumably lower during the TMS than of those tuned to the congruent direction, this behavioral result demonstrates that the offline TMS preferentially suppressed attributes encoded by the least active neural populations. In contrast to direction discrimination, motion detection was not impaired in a direction-specific manner. This shows that the requirements of the psychophysical task, in conjunction with the relative activity states of neuronal populations when TMS is applied, can be used to selectively interfere with overlapping neuronal populations.


Optimal use of docetaxel (Taxotere): maximizing its potential.
The safety of docetaxel (Taxotere) has been evaluated in the safety overview population consisting of 1070 patients recruited to phase II trials. These patients received a total of 4989 cycles of therapy (median four cycles per patient). Since docetaxel is known to be metabolized in the liver, hepatic impairment was predicted to be a risk factor for increased toxicity and was studied prospectively, comparing the 42 patients in the overview population with moderate hepatic impairment with the 1028 patients with liver function within normal limits. Hepatic dysfunction was associated with an increase in the percentage of cycles of therapy during which febrile neutropenia occurred and the number of patients suffering documented infection and severe (grade 3/4) stomatitis. The incidence of toxic death was also increased in patients with moderate hepatic impairment. The severity of fluid retention, a cumulative toxicity of docetaxel, was found to be reduced, and its onset delayed, by prophylactic treatment with corticosteroids for 5 days, starting 1 day before docetaxel administration. Treatment with corticosteroids was also recommended to reduce the incidence and severity of hypersensitivity reactions and cutaneous toxicities. The most frequent severe non-haematological toxicity of docetaxel was asthenia. Other non-haematological toxicities were generally mild or moderate.


CCK-B receptors produce similar signals but have opposite growth effects in CHO and Swiss 3T3 cells.
Rat cholecystokinin-B (CCK-B) receptors were transfected into Chinese hamster ovary (CHO)-K1 (CHO-CCK-B) and Swiss 3T3 (Swiss 3T3-CCK-B) cells, and the effects of receptor activation on cell proliferation and intracellular signaling were investigated. CCK octapeptide (CCK-8) treatment had no effect on cell growth in quiescent CHO-CCK-B cells but inhibited DNA synthesis, proliferation, and colony formation when the cells were grown in fetal bovine serum (FBS). In contrast, CCK-8 stimulated DNA synthesis in quiescent Swiss 3T3-CCK-B cells and had no effect when the cells were grown in FBS. These differences in growth responses were not due to differences in the level of receptor expression, as similar numbers of receptors were present in both cell types. To determine whether the different growth effects were due to differences in receptor coupling to common second messenger pathways, we investigated the effects of CCK-8 on several known intracellular signals. In both cell types, CCK-8 stimulated increases in intracellular Ca2+ concentration and polyphosphoinositide hydrolysis with similar potencies and efficacies. CCK-8 also stimulated arachidonate release from both cell types, although the potency was higher in the CHO cells. Adenosine 3',5'-cyclic monophosphate generation was observed at high agonist concentrations in both cell types and was much greater in cells with higher receptor density. In summary, receptor activation had opposite effects on growth parameters in CHO and Swiss 3T3 cells, but only minor differences were observed in the characteristics of CCK-B receptor coupling to specific second messengers in the two cell types. Thus cellular context is a principal determinant of the biological effects of CCK-B receptor activation, and differences in biological responses may occur independently of major differences in receptor coupling.


Metabolism of agmatine in macrophages: modulation by lipopolysaccharide and inhibitory cytokines.
Agmatine is an amine derived from the decarboxylation of arginine by arginine decarboxylase (ADC) and metabolized to putrescine by agmatinase. While prevalent in bacteria and plants, agmatine and its metabolic enzymes have been recently identified in mammalian tissues. In the present study we sought to determine: (a) whether macrophages (cell line RAW 264.7) express ADC and agmatinase, and (b) if the enzymes are regulated by lipopolysaccharide (LPS), and/or by the inhibitory cytokines transforming growth factor-beta (TGF-beta), interleukin-10 (IL-10) and interleukin-4 (IL-4). LPS induced a dose-dependent stimulation of agmatinase, while it decreased ADC, the effect in both cases being maximum at 20 h. As expected, LPS dose-dependently stimulated the inducible nitric oxide synthase activity (iNOS). A strong correlation was observed between the effects of LPS on the agmatine-related enzymes and iNOS. By contrast, exposure to IL-10 and TGF-beta caused a reduction in ADC and agmatinase, whereas IL-4 was ineffective on ADC, but reverted the LPS-induced increase of agmatinase. We conclude that the agmatine pathway may be an alternative metabolic route for arginine in macrophages, suggesting a regulatory role of agmatine during inflammation.


Management of acute hepatitis B.
Acute hepatitis B virus (HBV) is a common cause of acute icteric hepatitis in adults. The vast majority of these patients resolve this acute infection and develop long-lasting immunity. In contrast, the vast majority of patients who develop chronic HBV have minimal symptoms and do not develop jaundice after becoming infected with HBV. These patients will frequently remain undiagnosed for years or decades. Approximately 1% of persons with acute HBV develop acute liver failure. Preventing acute HBV with vaccination is the best treatment. Although universal vaccination is now administered to newborns in many countries, the majority of adults have not been vaccinated and remain at risk. Because the majority of patients with acute HBV resolve this infection spontaneously, treatment with an oral anti-HBV agent is not necessary. However, the use of an oral anti-HBV agent is not unreasonable to use in a patient who is developing acute liver failure from severe acute HBV.


[Monitoring quality of health care using indicators].
In a modern health care system continuing measurement and monitoring of relevant clinical data comprise the base of documentation for the quality of care. Internationally there has been increasing focus on monitoring and measuring the quality of care using principals from clinical epidemiology, including quality monitoring using clinical indicators. Indicators can be derived with regard to structure, process and outcome. The indicators should be in a form which can be used within the individual department and for comparison between hospitals nationally and internationally. The idea of professional indicators is to be able to track developments over time, as well to be able to respond at relevant times. The work with quality indicators will support continuing work with quality development in the medical specialities as well as giving a possibility for dialogue between politicians, the management system and the medical specialities about priority of resources and the structure of the hospital system, which extend further than the simpler issues of finance, which have been dominating until now.


Hypothermia does not influence liver damage and function in a porcine polytrauma model.
Previous studies revealed evidence that induced hypothermia attenuates ischemic organ injuries after severe trauma. In the present study, the effect of hypothermia on liver damage was investigated in a porcine long term model of multi-system injury, consisting of blunt chest trauma, penetrating abdominal trauma, musculoskeletal injury, and hemorrhagic shockMETHODS: In 30 pigs, a standardized polytrauma including blunt chest trauma, penetrating abdominal trauma, musculoskeletal injury, and hemorrhagic shock of 45% of total blood volume was induced. Following trauma, hypothermia of 33∘C was induced for 12 h and intensive care treatment was evaluated for 48 h. As outcome parameters, we assessed liver function and serum transaminase levels as well as a histopathological analysis of tissue samples. A further 10 animals served as controls. Serum transaminase levels were increased at the end of the observation period following hypothermia without reaching statistical significance compared to normothermic groups. Liver function was preserved (p⩽ 0.05) after the rewarming period in hypothermic animals but showed no difference at the end of the observation period. In H&E staining, cell death was slightly increased hypothermic animals and caspase-3 staining displayed tendency towards more apoptosis in hypothermic group as well. Induction of hypothermia could not significantly improve hepatic damage during the first 48 h following major trauma. Further studies focusing on multi-organ failure including a longer observation period are required to illuminate the impact of hypothermia on hepatic function in multiple trauma patients.


An LFA-1 (alphaLbeta2) small-molecule antagonist reduces inflammation and joint destruction in murine models of arthritis.
LFA-1 appears to play a central role in normal immune responses to foreign Ags. In autoimmune or inflammatory diseases, there is increased expression of LFA-1 and/or its counterligand, ICAM-1. Others have demonstrated that the targeted disruption of LFA-1:ICAM interactions, either by gene deletion or Ab treatment in mice, results in reduced leukocyte trafficking, inflammatory responses, and inhibition of inflammatory arthritis in the K/BxN serum transfer model. However, there has been little success in finding a small-molecule LFA-1 antagonist that can similarly impact rodent models of arthritis. In this paper, we present the first reported example of an LFA-1 small-molecule antagonist, BMS-587101, that is efficacious in preclinical disease models. In vitro, BMS-587101 inhibited LFA-1-mediated adhesion of T cells to endothelial cells, T cell proliferation, and Th1 cytokine production. Because BMS-587101 exhibits in vitro potency, cross-reactivity, and oral bioavailability in rodents, we evaluated the impact of oral administration of this compound in two different models of arthritis: Ab-induced arthritis and collagen-induced arthritis. Significant impact of BMS-587101 on clinical score in both models was observed, with inhibition comparable or better than anti-mouse LFA-1 Ab. In addition, BMS-587101 significantly reduced cytokine mRNA levels in the joints of Ab-induced arthritis animals as compared with those receiving vehicle alone. In paws taken from the collagen-induced arthritis study, the bones of vehicle-treated mice had extensive inflammation and bone destruction, whereas treatment with BMS-587101 resulted in marked protection. These findings support the potential use of an LFA-1 small-molecule antagonist in rheumatoid arthritis, with the capacity for disease modification.


Cutaneous leishmaniasis in an immigrant Saudi worker: a case report.
Cutaneous leishmaniasis (CL), an uncommon disorder in South-East Asia, including Bangladesh, often presents as granulomatous plaque on the exposed areas, with a high index of suspicion required for diagnosis. Here we report the first imported case of CL caused by Leishmania tropica in a migrant Bangladeshi worker in the Kingdom of Saudi Arabia (KSA). The case, initially suspected as a case of cutaneous tuberculosis, arrived at specimens reception unit (SRU) of diagnostic labs of icddr,b being referred by the physician for ALS testing for tuberculosis. At his arrival in the SRU, one of the health personnel of the unit who used to work in KSA suspected him as a case of CL. The diagnosis was confirmed by smear microscopy which revealed plenty of amastigotes within macrophages. PCR was performed to confirm the species. He was treated with sodium stibogluconate at Shahid Suhrawardy Medical College Hospital, Dhaka.


Hospital burden of pulmonary arterial hypertension in France.
Pulmonary arterial hypertension is a severe disease associated with frequent hospitalisations. This retrospective analysis of the French medical information PMSI-MSO database aimed to describe incident cases of patients with pulmonary arterial hypertension hospitalised in France in 2013 and to document associated hospitalisation costs from the national health insurance perspective. Cases of pulmonary arterial hypertension were identified using a diagnostic algorithm. All cases hospitalised in 2013 with no hospitalisation the previous two years were retained. All hospital stays during the year following the index hospitalisation were extracted, and classified as incident stays, monitoring stays or stays due to disease worsening. Costs were attributed from French national tariffs. 384 patients in France were hospitalised with incident pulmonary arterial hypertension in 2013. Over the following twelve months, patients made 1,271 stays related to pulmonary arterial hypertension (415 incident stays, 604 monitoring stays and 252 worsening stays). Mean age was 59.6 years and 241 (62.8%) patients were women. Liver disease and connective tissue diseases were documented in 62 patients (16.1%) each. Thirty-one patients (8.1%) died during hospitalisation and four (1.0%) received a lung/heart-lung transplantation. The total annual cost of these hospitalisations was € 3,640,382. € 2,985,936 was attributable to standard tariffs (82.0%), € 463,325 to additional ICU stays (12.7%) and € 191,118 to expensive drugs (5.2%). The mean cost/stay was € 2,864, ranging from € 1,282 for monitoring stays to € 7,285 for worsening stays. Although pulmonary arterial hypertension is rare, it carries a high economic burden.


Cross-diffusion-induced subharmonic spatial resonances in a predator-prey system.
In this paper we investigate the complex dynamics originated by a cross-diffusion-induced subharmonic destabilization of the fundamental subcritical Turing mode in a predator-prey reaction-diffusion system. The model we consider consists of a two-species Lotka-Volterra system with linear diffusion and a nonlinear cross-diffusion term in the predator equation. The taxis term in the search strategy of the predator is responsible for the onset of complex dynamics. In fact, our model does not exhibit any Hopf or wave instability, and on the basis of the linear analysis one should only expect stationary patterns; nevertheless, the presence of the nonlinear cross-diffusion term is able to induce a secondary instability: due to a subharmonic spatial resonance, the stationary primary branch bifurcates to an out-of-phase oscillating solution. Noticeably, the strong resonance between the harmonic and the subharmonic is able to generate the oscillating pattern albeit the subharmonic is below criticality. We show that, as the control parameter is varied, the oscillating solution (subT mode) can undergo a sequence of secondary instabilities, generating a transition toward chaotic dynamics. Finally, we investigate the emergence of subT-mode solutions on two-dimensional domains: when the fundamental mode describes a square pattern, subharmonic resonance originates oscillating square patterns. In the case of subcritical Turing hexagon solutions, the internal interactions with a subharmonic mode are able to generate the so-called "twinkling-eyes" pattern.


Monte Carlo modeling of the Elekta Versa HD and patient dose calculation with EGSnrc/BEAMnrc.
Numerous studies have proven the Monte Carlo method to be an accurate means of dose calculation. Although there are several commercial Monte Carlo treatment planning systems (TPSs), some clinics may not have access to these resources. We present a method for routine, independent patient dose calculations from treatment plans generated in a commercial TPS with our own Monte Carlo model using free, open-source software. A model of the Elekta Versa HD linear accelerator was developed using the EGSnrc codes. A MATLAB script was created to take clinical patient plans and convert the DICOM RTP files into a format usable by EGSnrc. Ten patients' treatment plans were exported from the Monaco TPS to be recalculated using EGSnrc. Treatment simulations were done in BEAMnrc, and doses were calculated using Source 21 in DOSXYZnrc. Results were compared to patient plans calculated in the Monaco TPS and evaluated in Verisoft with a gamma criterion of 3%/2 mm. Our Monte Carlo model was validated within 1%/1-mm accuracy of measured percent depth doses and profiles. Gamma passing rates ranged from 82.1% to 99.8%, with 7 out of 10 plans having a gamma pass rate over 95%. Lung and prostate patients showed the best agreement with doses calculated in Monaco. All statistical uncertainties in DOSXYZnrc were less than 3.0%. A Monte Carlo model for routine patient dose calculation was successfully developed and tested. This model allows users to directly recalculate DICOM RP files containing patients' plans that have been exported from a commercial TPS.


Effects of direct and averted gaze on the perception of facially communicated emotion.
Research has largely neglected the effects of gaze direction cues on the perception of facial expressions of emotion. It was hypothesized that when gaze direction matches the underlying behavioral intent (approach-avoidance) communicated by an emotional expression, the perception of that emotion would be enhanced (i.e., shared signal hypothesis). Specifically, the authors expected that (a) direct gaze would enhance the perception of approach-oriented emotions (anger and joy) and (b) averted eye gaze would enhance the perception of avoidance-oriented emotions (fear and sadness). Three studies supported this hypothesis. Study 1 examined emotional trait attributions made to neutral faces. Study 2 examined ratings of ambiguous facial blends of anger and fear. Study 3 examined the influence of gaze on the perception of highly prototypical expressions.


Multiple levels of control of the stage- and region-specific expression of rat intestinal lactase.
To elucidate the mechanisms leading to the functional regionalization of the digestive epithelium, lactase expression was analyzed at the protein, mRNA and gene levels, along the intestinal tract at various stages of the rat postnatal development. In the colon of neonates, the transient expression of mRNA and enzyme correlated well with gene transcription. In contrast to the colon, complex patterns were observed in the small intestine. In suckling animals, the mRNA was present at a high level despite the progressive decline of enzyme activity. Crypts were devoid of mRNA and the transcript mainly accumulated in the lower half of the villi. From weaning onwards, a functional regionalization of the epithelium was defined, characterized by the modification of the longitudinal distribution of lactase mRNA. Indeed the transcript remained abundant in the distal duodenum, jejunum and proximal ileum, but decreased in the proximal duodenum and became virtually absent in the distal ileum. Concomitantly, the mRNA and enzyme distribution along the villi changed in the different segments of the small intestine. Patterns similar to those described in sucklings were retained in the adult jejunum. In contrast, mRNA and enzyme could no longer be detected in the distal ileum, while mosaicism appeared in the proximal duodenum. In vitro transcription assays carried out with isolated nuclei suggested that the decay of lactase mRNA in the proximal duodenum at weaning was associated with a decreasing rate of transcription of the gene. However active gene transcription was retained in the nuclei of the adult jejunum and ileum. The loss of mRNA in the adult distal ileum despite the maintenance of active transcription did not result from an intragenic block of pre-RNA elongation, as shown by transcription assays carried out at various positions of the lactase gene. In addition, we looked for the ontogenic decline of lactase protein despite the maintenance of a high amount of mRNA in the jejunum, and it became evident that the fraction of mRNA present in polysomes was constant with age. Taken together, these data indicate that lactase constitutes an unusual marker of development and of functional regionalization of the intestinal tract which exhibits a complex time- and space-specific pattern of gene, mRNA, and protein expression. The distinct patterns occurring in the duodenum, jejunum, ileum, and the colon of pre- and postweaned rats depend on a combination of transcriptional, posttranscriptional, and posttranslational levels of regulation. and are associated with a different mRNA distribution along villi in each intestinal segment.


Wnt signaling and podocyte dysfunction in diabetic nephropathy.
Nephropathy is a major microvascular complication of diabetes mellitus and often leads to terminal renal failure in addition to contributing significantly to cardiovascular morbidity and mortality. Despites continuous advances, the pathogenesis of diabetic nephropathy remains poorly understood. Recent studies have underscored the significance of structural and functional changes in podocytes in the development and progression of diabetic nephropathy. The role of podocytes in health and diabetic nephropathy and abnormalities including podocyte hypertrophy, effacement, and apoptosis, and a detailed discussion on the role played by the Wnt-β-catenin signaling pathway in podocyte injury and dysfunction are the focus of this review. In addition, the role played by Wnt signaling in mediating the effects of known therapeutic strategies for diabetic nephropathy is also discussed.


Beta-adrenergic blockade and lipoprotein lipase activity in rat tissues after acute exercise.
The present experiments were aimed at evaluating the acute effects of exercise on lipoprotein lipase (LPL) activity in untrained rats. The activity of LPL was measured in postheparin plasma (PHP) before and at various times after a 1-h run on a treadmill (22 m/min, O degrees grade). LPL in PHP was 50% below pre-exercise levels immediately and 3 h after the run but was increased 65% over resting levels 24 h postexercise. To further characterize the very early fall in LPL activity in response to exercise and to assess the possible involvement therein of the beta-adrenergic pathway, LPL in heart, vastus lateralis muscle (VLM), and white (WAT) and brown (BAT) adipose tissues was determined at rest and immediately after exercise in rats that were treated or not with nadolol (25 mg.kg-1.day-1 for 30 days). Immediately after 1 h of exercise, there was a reduction in total enzyme activity in WAT (40% below resting levels), BAT (-58%), VLM (-53%), and heart (-30%). Exercise reduced serum triacylglycerol levels (-64%) and doubled those of nonesterified fatty acids. beta-Adrenergic blockade did not affect any of these variables. Both exercise and nadolol lowered serum cholesterol levels by approximately 20%, but the effects were not additive. These results show that the global intravascular pool of LPL undergoes divergent, time-dependent alterations in response to a single bout of moderate exercise. The acute downregulation of postheparin plasma LPL immediately after exercise reflected a fall in the total enzyme pool of all tissues studied.(ABSTRACT TRUNCATED AT 250 WORDS)


Sutures for anterior segment surgery.
A review of available suture materials in anterior segment surgery has been undertaken. The characteristics of each material and the procedures for which each may be useful have been described.


Economic evaluations and cost analyses in posttraumatic stress disorder: a systematic review.
Posttraumatic stress disorder is associated with a high economic burden. Costs of treatment are known to be high, and cost-effectiveness has been analysed for several treatment options. As no review on economic aspects of posttraumatic stress disorder exists, the aim of this study was to systematically review costs-of-illness studies and economic evaluations of therapeutic treatment for posttraumatic stress disorder, and to assess their quality. A systematic literature search was performed in March 2017 and was last updated in February 2020 in the databases PubMed, PsychInfo and NHS Economic Evaluation Database. Cost-of-illness studies and economic evaluations of treatment for posttraumatic stress disorder were selected. Extracted cost data were categorized as direct costs and indirect costs and inflated to 2015 US-$ purchasing power parities (PPP). Quality was assessed using an adapted cost-of-illness studies quality checklist, the Consensus on Health Economic Criteria list, and the questionnaire to assess relevance and credibility of modelling studies by the International Society for Pharmacoeconomics and Outcome Research. In total, 13 cost-of-illness studies and 18 economic evaluations were included in the review. Annual direct excess costs ranged from 512 US-$ PPP to 19,435 US-$ PPP and annual indirect excess costs were 5,021 US-$ PPP per person. Trauma-focused cognitive-behavioural therapy (+selective serotonin re-uptake inhibitor) was found to be cost-effective compared with treatment as usual and no treatment. Overall, included studies were of low and moderate quality. Studies used inappropriate economic study designs and lacked information on the economic perspective used. Posttraumatic stress disorder is a major public health problem that causes high healthcare costs. While trauma-focused cognitive-behavioural therapy was found to be cost-effective, further investigations regarding pharmacotherapy and other treatments are necessary.


GPR54 peptide agonists stimulate insulin secretion from murine, porcine and human islets.
This study was designed to determine the effects of 10 and 13 amino acid forms of kisspeptin on dynamic insulin secretion from mammalian islets since it is not clear from published data whether the shorter peptide is stimulatory while the longer peptide inhibits insulin release. Insulin secretion was measured by radioimmunoassay following perifusion of human, pig, rat and mouse isolated islets with kisspeptin-10 or kisspeptin-13 in the presence of 20 mM glucose. Both peptides stimulated rapid, reversible potentiation of glucose-stimulated insulin secretion from islets of all species tested. These data indicate that both kisspeptin-10 and kisspeptin-13, which is an extension of kisspeptin-10 by three amino acids, act directly at islet β-cells of various species to potentiate insulin secretion, and suggest that inhibitory effects reported in earlier studies may reflect differences in experimental protocols.


Falx and interhemispheric fissure on axial CT: II. Recognition and differentiation of interhemispheric subarachnoid and subdural hemorrhage.
Interhemispheric hyperdensity or unenhanced computed tomography was originally considered a sign of subarachnoid hemorrhage, the "falx sign." It has since been identified as a normal feature and has also been seen with interhemispheric subdural hemorrhage. To determine the differential features of interhemispheric hemorrhage, 50 patients with subarachnoid hemorrhage and 32 patients with interhemispheric subdural hematomas were reviewed. Subarachnoid hemorrhage produced anterior interhemispheric hyperdensity only, with a zigzag contour and extension from the calvarium to the rostrum of the corpus callosum. Interhemispheric subdural hematomas produce unilateral crescentic hyperdensities that are largest in the posterior superior part of the fissure, behind and above the splenium of the corpus callosum. Interhemispheric hyperdensity in children is more complex. Because the anterior part of the fissure is narrow in younger patients, subarachnoid hemorrhage may go undetected. Likewise, interhemispheric subdural hematomas in children are smaller and more difficult to recognize. They produce asymmetric thickening of the falx shadow with extension over the tentorium. They are, however, of great significance since they are generally seen in abused patients and carry a poor prognosis.


Cytotoxicity of 1,4-diamino-2-butanone, a putrescine analogue, to RKO cells: mechanism and redox imbalance.
α-Aminocarbonyl metabolites (e.g., 5-aminolevulinic acid and aminoacetone) and the wide spectrum microbicide 1,4-diamino-2-butanone (DAB) have been shown to exhibit pro-oxidant properties. In vitro, these compounds undergo phosphate-catalyzed enolization at physiological pH and subsequent superoxide radical-propagated aerobic oxidation, yielding a reactive α-oxoaldehyde and H2O2. DAB cytotoxicity to pathogenic microorganisms has been attributed to the inhibition of polyamine biosynthesis. However, the role played in cell death by reactive DAB oxidation products is still poorly understood. This work aims to clarify the mechanism of DAB-promoted pro-oxidant action on mammalian cells. DAB (0.05-10 mM) treatment of RKO cells derived from human colon carcinoma led to a decrease in cell viability (IC50 ca. 0.3 mM DAB, 24 h incubation). Pre-addition of either catalase (5 μM) or aminoguanidine (20 mM) was observed to partially inhibit the toxic effects of DAB to the cells, while N-acetyl-L-cysteine (NAC, 5 mM) or reduced glutathione (GSH, 5 mM) provided almost complete protection against DAB. Changes in redox balance and stress response pathways were indicated by the increased expression of HO-1, NQO1 and xCT. Moreover, the observation of caspase 3 and PARP cleavage products is consistent with DAB-triggered apoptosis in RKO cells, which was corroborated by the partial protection afforded by the pan-caspase inhibitor z-VAD-FMK. Finally, DAB treatment disrupted the cell cycle in response to increased p53 and activation of ATM. Altogether, these data support the hypothesis that DAB exerts cytotoxicity via a mechanism involving not only polyamine biosynthesis but also by DAB oxidation products.


Genotype-phenotype correlation in 440 patients with NPHP-related ciliopathies.
Nephronophthisis (NPHP), an autosomal recessive cystic kidney disease, is the most frequent genetic cause for end-stage renal failure in the first three decades of life. Mutations in 13 genes (NPHP1-NPHP11, AHI1, and CC2D2A) cause NPHP with ubiquitous expression of the corresponding proteins consistent with the multiorgan involvement of NPHP-related diseases. The genotype-phenotype correlation in these ciliopathies can be explained by gene locus heterogeneity, allelism, and the impact of modifier genes. In some NPHP-related ciliopathies, the nature of the recessive mutations determines disease severity. In order to define the genotype-phenotype correlation more clearly, we evaluated a worldwide cohort of 440 patients from 365 families with NPHP-related ciliopathies, in whom both disease-causing alleles were identified. The phenotypes were ranked in the order of severity from degenerative to degenerative/dysplastic to dysplastic. A genotype of two null alleles caused a range of phenotypes, with an increasing order of severity of NPHP1, NPHP3, NPHP4, NPHP5, NPHP2, NPHP10, NPHP6, to AHI1. Only NPHP6 showed allelic influences on the phenotypes; the presence of two null mutations caused dysplastic phenotypes, whereas at least one missense allele rescued it to a milder degenerative phenotype. We also found nine novel mutations in the NPHP genes. Thus, our studies have important implications for genetic counseling and planning of renal replacement therapy.


Predictors of Delayed Graft Function in Renal Transplantation.
This study aimed to analyze our data on delayed graft function (DGF) and to identify associated factors. This is a retrospective case-control study of all patients transplanted in our center over a period of 11 years (January 1, 2003, to December 31, 2014) comparing patients with immediate graft function (n = 332) to those with DGF (n = 165). DGF was defined as the need for hemodialysis within the first 7 days after transplantation. Donor and recipient characteristics as well as procedural factors were compared by univariate and multivariate logistic regression analyses. Overall, 33% of patients had DGF. The rate of DGF declined from 2003 to 2011. In cases with DGF, donors and recipients were significantly older (p = 0.004 and p = 0.005, respectively), had longer cold ischemia times (p = 0.039), more revision surgeries (p < 0.001), and more HLA mismatches (p = 0.001), especially in the DR locus (p = 0.002). Neither donor nor recipient gender, waiting time, nor CMV status had any influence. In multivariable analysis, significant risk factors were ischemia time and mismatches at the HLA-DR loci. DGF is a common complication in renal transplantation which occurred in 33% of our cases. Important factors identified were donor and recipient age, ischemia time, HLA mismatching, and revision surgery.


Invited commentary: Intrauterine epidemiology.
Traditionally, the investigation of preterm birth has relied on diagnostic definitions derived from maternal clinical presentation. However, clinical presentation may be only tangentially related to the underlying etiology of a disease. The utilization of data derived directly from the intrauterine or maternal systemic environment would be invaluable in consideration of the causes of preterm birth. In this issue, Kelly et al. (Am J Epidemiol. 2009;170(2)148-158) contribute to our understanding of the epidemiology of the intrauterine environment by classifying the vascular biology of the maternal-placental interface in cases of preterm delivery. Their histology-based approach observes that vascular conditions may be grouped into 5 constructs with specific relations to maternal and fetal vascular pathology. The frequencies of these constructs vary with regard to delivery indication and gestational age, suggesting that the intrauterine conditions associated with preterm birth are more complicated than originally appreciated. This work is laborious, and replication of the technique will be important. However, these authors have taken a large step toward introducing an "intrauterine" perspective into perinatal epidemiology and into our understanding of the underlying etiologies of preterm birth.


A new method of self-assessment during vocational training.
The ability to assess oneself in general practice has to be learned. The ideal time to develop this ability is during vocational training. The lessons learned may be some of the most important before entering general practice itself.As a new approach to this problem, a self-assessment book has been devised. The book has potential as a research tool and helps trainees to think more closely about their particular problems during patient contact.


A YAC contig spanning the blepharophimosis-ptosis-epicanthus inversus syndrome and propionic acidemia loci.
Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is an autosomal dominant condition consisting of congenital dysplasia of the eyelids with a reduced horizontal diameter of the palpebral fissures, droopy eyelids and epicanthus inversus. Two clinical entities have been described: type I and type II. The former is distinguished by female infertility, whereas the latter presents without other symptoms. Both type I and type II were recently mapped on the long arm of chromosome 3 (3q22-q23), suggesting a common gene may be affected. The centromeric and the telomeric limits of this region are well defined between loci D3S1316 and D3S1615, which reside approximately 5 cM apart. Here, we present the construction of a YAC contig spanning the entire BPES locus using 17 polymorphic markers, 2 STS and 28 ESTs. This region of approximately 5 Mb was covered by 31 YACs, and was supported by detailed FISH analysis. In addition, we have precisely mapped the propionyl-CoA carboxylase beta polypeptide (PCCB), the gene mutated in propionic acidemia, within this contig. Apart from providing a framework for the identification of the BPES gene, this contig will also be useful for the future identification of defects and genes mapped to this region, and for developing template resources for genomic sequencing.


The epidemiology of head injuries in Sweden from 1987 to 2000.
The purpose of the present study was to evaluate the variability in the annual head injury incidence rate in Sweden from 1987 to 2000. It was hypothesized that the annual incidence rate would decrease over time due to a variety of primary preventive strategies that have been introduced in Swedish society. We used the Hospital Discharge Register at the National Board for Health and Welfare and head injury codes 800-804, and 850-854 from ICD9 system and S2.0-S2.9, and S6.0-S6.9 codes from ICD-10 system. We evaluated the patterns of age, gender, external cause of injury (E-code), type of injury, length of hospital stay, and trends over time. Head injuries due to transportation collision were reduced over the 14-year period analysis. Falls persisted as the dominant cause of head injury. Overall, men had 2.1 times the incidence of head injury compared to women. There was a decline in younger ages experiencing a head injury over this interval, while the number of head injuries among elderly people increased over time. Concussion was about three times more frequent than fractures. Hematoma and diffuse or focal contusions had a much lower incidence rate than concussion. Concussions and fractures decreased over time. Diffuse or focal injuries showed a steady rate of occurrence over the study interval while hematoma increased. Although length of hospital stay varied widely from zero to more than 50 days, 73.6% of hospital days were confined to two days or less. The incidence rate is stable over this time frame. While head injuries attributable to transportation accidents decreased, falls made up an increasing proportion of head injuries. Since we observed an increase in head injuries among elderly, primary prevention strategies may need to be targeted at this age group, and at preventing falls.