Identification and Characteristics of Fusion Peptides Derived From Enveloped Viruses.
Membrane fusion events allow enveloped viruses to enter and infect cells. The study of these processes has led to the identification of a number of proteins that mediate this process. These proteins are classified according to their structure, which vary according to the viral genealogy. To date, three classes of fusion proteins have been defined, but current evidence points to the existence of additional classes. Despite their structural differences, viral fusion processes follow a common mechanism through which they exert their actions. Additional studies of the viral fusion proteins have demonstrated the key role of specific proteinogenic subsequences within these proteins, termed fusion peptides. Such peptides are able to interact and insert into membranes for which they hold interest from a pharmacological or therapeutic viewpoint. Here, the different characteristics of fusion peptides derived from viral fusion proteins are described. These criteria are useful to identify new fusion peptides. Moreover, this review describes the requirements of synthetic fusion peptides derived from fusion proteins to induce fusion by themselves. Several sequences of the viral glycoproteins E1 and E2 of HCV were, for example, identified to be able to induce fusion, which are reviewed here.


Integrated evaluation of a panel of neurochemical biomarkers to optimize diagnosis and prognosis in amyotrophic lateral sclerosis.
This study was undertaken to determine the diagnostic and prognostic value of a panel of serum biomarkers and to correlate their concentrations with several clinical parameters in a large cohort of patients with amyotrophic lateral sclerosis (ALS). One hundred forty-three consecutive patients with ALS and a control cohort consisting of 70 patients with other neurodegenerative disorders (DEG), 70 patients with ALS mimic disorders (ALSmd), and 45 healthy controls (HC) were included. Serum neurofilament light chain (NfL), ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1), glial fibrillary acidic protein (GFAP), and total tau protein levels were measured using ultrasensitive single molecule array. NfL correlated with disease progression rate (p < 0.001) and with the measures of upper motor neuron burden (p < 0.001). NfL was higher in the ALS patients with classic and pyramidal phenotype. GFAP was raised in ALS with cognitive-behavioral impairment compared with ALS with normal cognition. NfL displayed the best diagnostic performance in discriminating ALS from HC (area under the curve [AUC] = 0.990), DEG (AUC = 0.946), and ALSmd (AUC = 0.850). UCHL1 performed well in distinguishing ALS from HC (AUC = 0.761), whereas it was not helpful in differentiating ALS from DEG and ALSmd. In multivariate analysis, NfL (p < 0.001) and UCHL1 (p = 0.038) were independent prognostic factors. Survival analysis combining NfL and UCHL1 effectively stratified patients with lower NfL levels (p < 0.001). NfL is a useful biomarker for the diagnosis of ALS and the strongest predictor of survival. UCHL1 is an independent prognostic factor helpful in stratifying survival in patients with low NfL levels, likely to have slowly progressive disease. GFAP reflects extramotor involvement, namely cognitive impairment or frontotemporal dementia.


Growth hormone-releasing hormone receptor antagonists modify molecular machinery in the progression of prostate cancer.
Therapeutic strategies should be designed to transform aggressive prostate cancer phenotypes to a chronic situation. To evaluate the effects of the new growth hormone-releasing hormone receptor (GHRH-R) antagonists: MIA-602, MIA-606, and MIA-690 on processes associated with cancer progression as cell proliferation, adhesion, migration, and angiogenesis. We used three human prostate cell lines (RWPE-1, LNCaP, and PC3). We analyzed several molecules such as E-cadherin, β-catenin, Bcl2, Bax, p53, MMP2, MMP9, PCNA, and VEGF and signaling mechanisms that are involved on effects exerted by GHRH-R antagonists. GHRH-R antagonists decreased cell viability and provoked a reduction in proliferation in LNCaP and PC3 cells. Moreover, GHRH-R antagonists caused a time-dependent increase of cell adhesion in all three cell lines and retarded the wound closure with the highest value with MIA-690 in PC3 cells. GHRH-R antagonists also provoked a large number of cells in SubG0 phase revealing an increase in apoptotic cells in PC3 cell line. Taken all together, GHRH-R antagonists of the MIAMI series appear to be inhibitors of tumor progression in prostate cancer and should be considered for use in future therapeutic strategies on this malignancy.


[The oncologist and his patient].
The motifs of medical practise, in particular by the oncologist and the basic experience of the patient notably the cancer patient are presented. They represent the prerequisite for an individually taylored, solid relationship between physician and patient founding on mutual trust. Medical practise is nowadays founded on a scientific rationale. In contrast to earlier times the ethos of compassion pertains the private sphere. It is the physicians individual choice to proffer the traditional compassion as interest in the fate of an individual patient or not. The fundamental experience of an individual that his destiny lies no longer in his own hands expresses itself in a paradigmatic way by the perception of his illness. The cancer patient experiences--as other patients do--the threats but also his healing as occurrences not caused by him. He therefore does not identify his disease with objective impairments or pathologic findings. On the contrary, he perceives it as a threat to himself, his existence and his personality. The patient experiences his fate, to depend on circumstances and particularly on other individuals, their benevolence, their acceptance, their help. Trust is the basis for relationship between the cancer patient and his physician. Trust is also acceptance of dependence. Dependence is only acceptable if the physician is trustworthy. Trustworthiness cannot be gained by scientific competence alone but requires personal affection by compassion with the patients fate. The physician needs ethical competence to fulfill his task. Information of the patient gains a particular importance in this context. It, as well as the divergent assessment of the disease by patient and physician--the anthropologic difference--and the "equilibrating systems" are exposed.


The effects of saccharin on the development of neoplastic lesions initiated with N-methyl-N-nitrosourea in the rat urothelium.
Saccharin has been reported to induce urinary bladder tumors in multigeneration rat feeding studies and to promote bladder carcinogenesis in rats initiated with known bladder carcinogens. To examine the dose-dependent effects of saccharin on tumor promotion, sodium saccharin was administered at six levels in the diet (5.0, 2.5, 1.0, 0.5, 0.1, and 0%) to female Sprague-Dawley rats which had received, by trans-urethral instillation into the bladder, either a single dose of saline or an initiating dose of N-methyl-N-nitrosourea (MNU), a potent direct-acting carcinogen. Additional groups with and without MNU treatment received sodium saccharin (2%) in the drinking water, acid-saccharin (5%) in the diet, or MNU, four weekly doses, as a positive control. Histopathologic examination of the urinary bladders from dead and moribund animals and from animals sacrificed after 102 weeks on dose was performed, and benign papillomas were commonly observed in those animals given MNU. A statistical analysis of the lesions indicated an increase in tumor incidence and a decrease in time to tumor with increasing saccharin dose in dead and moribund animals. This response was observed in the dose series of 0 to 2.5% saccharin in the diet. Dead and moribund animals which had received 5% sodium saccharin exhibited few tumors. An increasing incidence of tumors in MNU-treated control animals was observed during the final weeks of the study. Although this increase in background tumors in senescent animals complicated the interpretation of the total tumor incidences, the results in dead and moribund animals (about 60% of the total) indicated that saccharin served as a tumor promoter in this two-stage carcinogenesis model system by decreasing the latency period of the lesions.


Prostaglandin endoperoxide analogues stimulate phospholipase C and protein phosphorylation during platelet shape change.
We have studied the effects of two stable prostaglandin endoperoxide analogues on platelet lipid metabolism in relation to specific platelet functional changes. During platelet shape change, the endoperoxide analogues induce the formation of 1,2-diacylglycerol and phosphatidic acid, indicating the activation of a phosphoinositide-specific phospholipase C. In parallel, they stimulate the phosphorylation of a 40-kd and a 20-kd protein. During platelet shape change, arachidonic acid is released, but not metabolized by platelet cyclo-oxygenase or lipoxygenase. Phospholipase C activation and platelet shape change are independent of extracellular Ca++ and Mg++, arachidonate metabolism, and release of adenosine diphosphate (ADP). Activation of phospholipase C during platelet aggregation seems, however, to be mediated partly by release of ADP. We conclude that endoperoxide analogues initially stimulate in platelets the formation of products derived from phospholipase C activation, which might serve as intracellular messengers for phosphorylation of specific proteins related to platelet shape change.


Photo-induced 6π-electrocyclisation and cycloreversion of isolated dithienylethene anions.
The diarylethene chromophore is commonly used in light-triggered molecular switches. The chromophore undergoes reversible 6π-electrocyclisation (ring closing) and cycloreversion (ring opening) reactions upon exposure to UV and visible light, respectively, providing bidirectional photoswitching. Here, we investigate the gas-phase photoisomerisation of meta- (m) and para- (p) substituted dithienylethene carboxylate anions (DTE-) using tandem ion mobility mass spectrometry coupled with laser excitation. The ring-closed forms of p-DTE- and m-DTE- are found to undergo cycloreversion in the gas phase with maximum responses associated with bands in the visible (λmax ≈ 600 nm) and the ultraviolet (λmax ≈ 360 nm). The ring-open p-DTE- isomer undergoes 6π-electrocyclisation in the ultraviolet region at wavelengths shorter than 350 nm, whereas no evidence is found for the corresponding electrocyclisation of ring-open m-DTE-, a situation attributed to the fact that the antiparallel geometry required for electrocyclisation of m-DTE- is energetically disfavoured. This highlights the influence of the carboxylate substitution position on the photochemical properties of DTE molecules. We find no evidence for the formation in the gas phase of the undesirable cyclic byproduct, which causes fatigue of DTE photoswitches in solution.


[Hereditary endocrine tumour diseases].
The paper reviews inheritable endocrine tumour syndromes based on recent literature and the experience of the authors. A number of inheritable endocrine tumour syndromes are caused by mutations in proto-oncogenes or tumour suppressor genes. These include multiple endocrine neoplasia (MEN), familial non-MEN neoplasia, Von Hippel-Lindau's disease and familial medullary thyroid carcinoma, pheochromocytoma and paraganglioma. These conditions are rare, but since inheritance is dominant with a near-complete penetrance, a number of carriers with a high risk of developing tumours are regularly found in the families of affected individuals. Identification of carriers with a high risk of developing tumours gives the opportunity to cure disease manifestations at an early stage and to avoid life-threatening complications. Family members who are not carriers can be excluded from follow up. DNA analyses of the most common disease genes are now performed in genetic laboratories in Norway.


Synthesis of Functionalized Arylacetamido-2-pyridones through ortho-C(sp2)-H-Activated Installation of Olefins and Alkynes.
We have identified different N-substituted 2-pyridones as inbuilt directing groups for selective C-H-activated functionalization instead of deprotecting and/or throwing away the directing groups. A robust general method for external ligand-free PdII-catalyzed C(sp2)-H olefination and alkynylation is established to access valuable phenylacetamido-2-pyridones. Diverse substrate scope has been demonstrated with 48 different examples with high yield and gram-scale synthesis. Adequate tolerance of valuable functional groups was also observed, such as olefins possessing esters, sulfone, amide, cyanide, and ketones, aromatic residues containing fluorine, chlorine, bromine, NO2, methyl, dimethyl, and methoxy, as well as 2-pyridone-N-bearing methyl, cyclopropyl methyl, cyclopentyl methyl, benzyl, phenyl, acetate, and acetamide groups, which smoothly produced the respective desired products. We used triisopropyl silane-substituted alkynes for the alkynylation reaction, which can easily be converted to several functional groups including terminal alkyne, heterocycle through click reaction, and others. Implementing our protocol, we have also demonstrated late-stage olefination and alkynylation of 2-pyridone, containing the CB2 agonist-type molecule with excellent yield. Considering N-substituted 2-pyridone acts as a biologically-active structural unit, this general method has the significance in terms of late-stage functionalization to access new molecular entities which can be employed in medicinal chemistry research through diverse C-H activation.


Targeting vulnerable groups of health poverty alleviation in rural China- what is the role of the New Rural Cooperative Medical Scheme for the middle age and elderly population?
In light of the health poverty alleviation policy, we explore whether the New Rural Cooperative Medical System (NRCMS) has effectively reduced the economic burden of medical expenses on rural middle-aged and elderly people and other impoverished vulnerable groups. The study aims to provide evidence that can be used to improve the medical insurance system. Data were obtained from the 2015 China Health and Retirement Longitudinal Study (CHARLS). The method of calculating the catastrophic health expenditure (CHE) and impoverishment by medical expense (IME) was adopted from the World Health Organization (WHO). The treatment effect model was used to identify the determinants of CHE for rural middle-aged and elderly people. The incidence of CHE in rural China for middle-aged and elderly people is 21.8%, and the IME is 8.0%. The households that had enrolled in the NRCMS suffered higher CHE (21.9%) and IME (8.0%), than those that had not enrolled (CHE: 20.6% and IME: 7.7%). The NRCMS did not provide sufficient economic protection from CHE for households with three or more chronic diseases, inpatients, or households with members aged over 65 years. Key risk factors for the CHE included education levels, households with inpatients, households with members aged over 65 years, and households with disabilities. Although the NRCMS has reduced barriers to the usage of household health services by reducing people's out-of-pocket payments, it has not effectively reduced the risk of these households falling into poverty. Our research identifies the characteristics of vulnerable groups that the NRCMS does not provide enough support for, and which puts them at a greater risk of falling into poverty due to health impoverishment.


An additional GerE-controlled gene encoding an abundant spore coat protein from Bacillus subtilis.
We describe the identification and characterization of a gene, herein designated cotG, encoding an abundant coat protein from the spores of Bacillus subtilis. The cotG open reading frame is 195 codons in length and is capable of encoding a polypeptide of 24 kDa that contains nine tandem copies of the 13-amino-acid long, approximately repeated sequence H/Y-K-K-S-Y-R/C-S/T-H/Y-K-K-S-R-S. cotG is located at 300 degrees on the genetic map close to another coat protein gene, cotB. The cotG and cotB genes are in divergent orientation and are separated by 1.3 kb. Like the promoter for cotB, the cotG promoter is induced at a late stage of sporulation under the control of the RNA polymerase sigma factor sigma K and the DNA-binding protein GerE. The -10 and -35 nucleotide sequences of the cotG promoter resemble those of other promoters recognized by sigma K-containing RNA polymerase, and centered 70 bp upstream of the apparent start site is a sequence that matches the consensus binding site for GerE. Spore coat proteins from a newly constructed cotG null mutant lack not only CotG but also CotB, a finding that suggests that CotG may be a morphogenetic protein that is required for the incorporation of CotB into the coat.


Pediatric neuroimaging in early childhood and infancy: challenges and practical guidelines.
Structural and functional magnetic resonance imaging (fMRI) has been used increasingly to investigate typical and atypical brain development. However, in contrast to studies in school-aged children and adults, MRI research in young pediatric age groups is less common. Practical and technical challenges occur when imaging infants and children, which presents clinicians and research teams with a unique set of problems. These include procedural difficulties (e.g., participant anxiety or movement restrictions), technical obstacles (e.g., availability of child-appropriate equipment or pediatric MR head coils), and the challenge of choosing the most appropriate analysis methods for pediatric imaging data. Here, we summarize and review pediatric imaging and analysis tools and present neuroimaging protocols for young nonsedated children and infants, including guidelines and procedures that have been successfully implemented in research protocols across several research sites.


Bankruptcy as Implicit Health Insurance.
This paper examines the implicit health insurance that households receive from the ability to declare bankruptcy. Exploiting multiple sources of variation in asset exemption law, I show that uninsured households with a greater financial cost of bankruptcy make higher out-of-pocket medical payments, conditional on the amount of care received. In turn, I find that households with greater wealth at risk are more likely to hold health insurance. The implicit insurance from bankruptcy distorts the insurance coverage decision. Using a microsimulation model, I calculate that the optimal Pigovian penalties are three-quarters as large as the average penalties under the Affordable Care Act.


Trends in adverse events and related health-care facility utilization from cough and cold medication exposures in children.
Initial research following regulatory changes addressing the pediatric safety of cough and cold medications (CCMs) demonstrated decreases in adverse events (AEs). Using a national multi-source surveillance system, we studied subsequent CCM-related AE case rate trends and associated health-care facility (HCF) evaluation in children. Data were collected from 2009 to 2016. Case eligibility included: age <12 years; exposure to an over-the-counter product containing ≥1 CCM pharmaceutical ingredient; ≥1 significant AE that occurred in the United States. About 4756 (72.6%) cases were determined at least potentially related to an index ingredient. Accidental unsupervised ingestions (AUIs; 3134; 65.9%) were the most common case type. Nearly half of AE cases involved children 2 to <4 years old (2,159; 45.4%). The AE case rate did not change significantly over time (p = 0.22). The proportion of AE cases resulting in HCF admission increased from 32.4% (207) in 2009 to 43.4% (238) in 2016 (p < 0.01). Exposures to diphenhydramine (1,305; 67.3%) and/or dextromethorphan (591; 30.5%) were involved in the majority of HCF admissions. The proportion of AE cases resulting in HCF admission increased from 2009 to 2016. Efforts to prevent AUIs such as packaging innovation and engineering controls, particularly for diphenhydramine and dextromethorphan-containing products, should be pursued.


Changes in the protein nutritional status of adolescent wrestlers.
The protein nutritional status of adolescent wrestlers was studied to determine whether changes occur during a season of competition and weight loss. Subjects (N = 18) were measured prior to the start of the season (PRE), twice in the midseason, and once during late season (LATE) for weight, percent body fat, and height. At each of these times, a venous blood sample was obtained from the subjects, who were fasted, and analyzed for concentrations of albumin, prealbumin, retinol binding protein (RBP), blood urea nitrogen (BUN), hemoglobin, hematocrit, and 23 amino acids. Diet records were kept by subjects to assess daily energy, protein, fat, and carbohydrate intake. Data were analyzed by repeated measures ANOVA. Results showed that wrestlers decreased weight by an average of 6.6 +/- 0.9% and that percent body fat, fat-free weight, plasma levels of prealbumin and RBP, the ratio of total essential amino acids to total amino acids, and dietary energy nutrient intakes were significantly lower at LATE compared to PRE. RBP decreased during midseason and averaged (+/- SE) 3.21 +/- 0.15 mg.100 ml-1 at LATE; prealbumin was significantly lower at LATE with a mean value of 19.8 +/- 1.0 mg.100 ml-1. Total energy intake decreased from PRE values by 35%, to approximately 27 kcal.kg-1.d-1 during the season. In conclusion, in these high school wrestlers who lost approximately 6.6% of weight, there were adverse effects on some of the indices of protein nutritional status.


A knowledge-based system for assisted ventilation of patients in intensive care units.
The procedure for weaning a patient with respiratory insufficiency from mechanical ventilation may be complex and requires expertise obtained by long clinical practice. We designed a knowledge-based system for the management of patients receiving respiratory support and implemented a weaning procedure. The system is intended for patients whose spontaneous respiratory activity is assisted by a Hamilton Veolar ventilator delivering a positive pressure plateau during inspiration (Pressure Support Ventilation mode). Our closed-loop real-time system running on a Personal Computer continuously adapts the assistance provided by the ventilator to the patient's evolution, and indicates when the patient can be withdrawn from the ventilator. Three parameters are used to appreciate the 'respiratory comfort' of the patient: breathing frequency, which we consider the most informative index, tidal volume and end-tidal CO2 pressure. A preliminary study of 19 patients was performed to evaluate the ability of our system to adapt the assistance to the patient's needs, with the main objective of facilitating weaning by gradually lowering the level of assistance. In 10 of these patients, considered as good candidates for weaning on the strength of objective criteria, the system maintained the breathing pattern in a zone of comfort for 95% of the period of assisted ventilation and stated that they were 'weanable'. This was consistent with the clinical evolution of all 10 patients. These results show that such a system can provide effective management for mechanically ventilated patients.


Current status of and future issues related to endoscopic papillectomy.
Endoscopic papillectomy is widely performed to treat duodenal papillary tumors, particularly at high-volume centers. It is indicated for adenomas without intraductal extension of the bile or pancreatic ducts. However, despite numerous reports of carcinomas that expand the indications to include well-differentiated adenocarcinomas that do not invade the sphincter of Oddi, the low agreement between biopsy and final pathological diagnosis, as well as the current inability of imaging modalities to diagnose sphincter of Oddi invasion, makes it difficult to consider expanding indications. Although complications can be prevented by certain methods, such as pancreatic duct stenting, and the frequency of severe complications has decreased, the safety of the procedure remains unconfirmed. In the future, this technology is expected to progress and enable wider applications, including those in tumors with extensive horizontal spread and those with intraductal extension of the bile and pancreatic ducts. Such technology may also improve the safety and accuracy of diagnosis.


Protein topology classification using two-stage support vector machines.
The determination of the first 3-D model of a protein from its sequence alone is a non-trivial problem. The first 3-D model is the key to the molecular replacement method of solving phase problem in x-ray crystallography. If the sequence identity is more than 30%, homology modelling can be used to determine the correct topology (as defined by CATH) or fold (as defined by SCOP). If the sequence identity is less than 25%, however, the task is very challenging. In this paper we address the topology classification of proteins with sequence identity of less than 25%. The input information to the system is amino acid sequence, the predicted secondary structure and the predicted real value relative solvent accessibility. A two stage support vector machine (SVM) approach is proposed for classifying the sequences to three different structural classes (alpha, beta, alpha+beta) in the first stage and 39 topologies in the second stage. The method is evaluated using a newly curated dataset from CATH with maximum pairwise sequence identity less than 25%. An impressive overall accuracy of 87.44% and 83.15% is reported for class and topology prediction, respectively. In the class prediction stage, a sensitivity of 0.77 and a specificity of 0.91 is obtained. Data file, SVM implementation (SVMHEAVY) and result files can be downloaded from http://www.ee.unimelb.edu.au/ISSNIP/downloads/.


Stress-induced transcription of satellite III repeats.
Exposure of mammalian cells to stress induces the activation of heat shock transcription factor 1 (HSF1) and the subsequent transcription of heat shock genes. Activation of the heat shock response also correlates with a rapid relocalization of HSF1 within a few nuclear structures termed nuclear stress granules. These stress-induced structures, which form primarily on the 9q12 region in humans through direct binding of HSF1 to satellite III repeats, do not colocalize with transcription sites of known hsp genes. In this paper, we show that nuclear stress granules correspond to RNA polymerase II transcription factories where satellite III repeats are transcribed into large and stable RNAs that remain associated with the 9q12 region, even throughout mitosis. This work not only reveals the existence of a new major heat-induced transcript in human cells that may play a role in chromatin structure, but also provides evidence for a transcriptional activity within a locus considered so far as heterochromatic and silent.


Molecular characterization of adeno-associated viruses infecting children.
Although adeno-associated virus (AAV) infection is common in humans, the biology of natural infection is poorly understood. Since it is likely that many primary AAV infections occur during childhood, we set out to characterize the frequency and complexity of circulating AAV isolates in fresh and archived frozen human pediatric tissues. Total cellular DNA was isolated from 175 tissue samples including freshly collected tonsils (n = 101) and archived frozen samples representing spleen (n = 21), lung (n = 16), muscle (n = 15), liver (n = 19), and heart (n = 3). Samples were screened for the presence of AAV and adenovirus sequences by PCR using degenerate primers. AAV DNA was detected in 7 of 101 (7%) tonsil samples and two of 74 other tissues (one spleen and one lung). Adenovirus sequences were identified in 19 of 101 tonsils (19%), but not in any other tissues. Complete capsid gene sequences were recovered from all nine AAV-positive tissues. Sequence analyses showed that eight of the capsid sequences were AAV2-like (approximately 98% amino acid identity), while the single spleen isolate was intermediate between serotypes 2 and 3. Comparison to the available AAV2 crystal structure revealed that the majority of the amino acid substitutions mapped to surface-exposed hypervariable domains. To further characterize the AAV capsid structure in these samples, we used a novel linear rolling-circle amplification method to amplify episomal AAV DNA and isolate infectious molecular clones from several human tissues. Serotype 2-like viruses were generated from these DNA clones and interestingly, failed to bind to a heparin sulfate column. Inspection of the capsid sequence from these two clones (and the other six AAV2-like isolates) revealed that they lacked arginine residues at positions 585 and 588 of the capsid protein, which are thought to be essential for interaction with the heparin sulfate proteoglycan coreceptor. These data provide a framework with which to explore wild-type AAV persistence in vivo and provide additional tools to further define the biodistribution and form of AAV in human tissues.


Calibration of Traceable Solid Mock (131)I Phantoms Used in an International SPECT Image Quantification Comparison.
The International Atomic Energy Agency (IAEA) has organized an international comparison to assess Single Photon Emission Computed Tomography (SPECT) image quantification capabilities in 12 countries. Iodine-131 was chosen as the radionuclide for the comparison because of its wide use around the world, but for logistical reasons solid (133)Ba sources were used as a long-lived surrogate for (131)I. For this study, we designed a set of solid cylindrical sources so that each site could have a set of phantoms (having nominal volumes of 2 mL, 4 mL, 6 mL, and 23 mL) with traceable activity calibrations so that the results could be properly compared. We also developed a technique using two different detection methods for individually calibrating the sources for (133)Ba activity based on a National standard. This methodology allows for the activity calibration of each (133)Ba source with a standard uncertainty on the activity of 1.4 % for the high-level 2-, 4-, and 6-mL sources and 1.7 % for the lower-level 23 mL cylinders. This level of uncertainty allows for these sources to be used for the intended comparison exercise, as well as in other SPECT image quantification studies.


Does combination of estradiol and sesame oil improve the oocyte quality, embryo development and expressions of Zp3, E-cad, and Ctnnb1 genes in mice? An experimental study.
Aging may reduce oocyte maturation, embryo quality, and fertility potential. To compare the effect of estradiol (E2) and sesame oil on oocyte and embryo quality between young and old mice. Sixty old and young female mice were divided in to two groups (30 mice/group, grouped by age). Each group was divided into three subgroups of mice treated with sesame oil, E2 + sesame oil, and normal saline as control group. After ovulation induction, some oocytes were considered for in vitro fertilization and the rest were assessed for morphological status. After obtaining the two-cell embryos, the embryos were collected to determine the expression of zona pellucida (ZP) glycoprotein 3, E-cadherin, and β-catenin genes and some of them followed until the blastocysts stage to evaluate the viability. The findings showed that the mean ZP and perivitelline space thickness increased in the old mice that received the E2 + sesame oil treatment. The number of 2-cell embryos, blastocysts, and live cells were significantly higher in the old group treated with sesame oil respectively (p = 0.018, 0.002, and < 0.0001, respectively). The normal ZP shape and refractile body numbers increased in the old mice that were treated with sesame oil, respectively. The E-cadherin gene was downregulated in the treatment groups compared to the controls. Sesame oil showed a better response in the old mice, because aging is associated with an increased rate of reactive oxygen species, causing deficiencies in both oocyte and embryo qualities.


Estimation of admixture proportions: a likelihood-based approach using Markov chain Monte Carlo.
When populations are separated for long periods and then brought into contact for a brief episode in part of their range, this can result in genetic admixture. To analyze this type of event we considered a simple model under which two parental populations (P1 and P2) mix and create a hybrid population (H). After that event, the three populations evolve under pure drift without exchange during T generations. We developed a new method, which allows the simultaneous estimation of the time since the admixture event (scaled by the population size t(i) = T/N(i), where N(i) is the effective population size of population i) and the contribution of one of two parental populations (which we call p1). This method takes into account drift since the admixture event, variation caused by sampling, and uncertainty in the estimation of the ancestral allele frequencies. The method is tested on simulated data sets and then applied to a human data set. We find that (i) for single-locus data, point estimates are poor indicators of the real admixture proportions even when there are many alleles; (ii) biallelic loci provide little information about the admixture proportion and the time since admixture, even for very small amounts of drift, but can be powerful when many loci are used; (iii) the precision of the parameters' estimates increases with sample size n = 50 vs. n = 200 but this effect is larger for the t(i)'s than for p1; and (iv) the increase in precision provided by multiple loci is quite large, even when there is substantial drift (we found, for instance, that it is preferable to use five loci than one locus, even when drift is 100 times larger for the five loci). Our analysis of a previously studied human data set illustrates that the joint estimation of drift and p1 can provide additional insights into the data.


Bacillus ruris sp. nov., from dairy farms.
Four novel ellipsoidal spore-forming Bacillus isolates with swollen sporangia, isolated from raw milk and feed concentrate, showed a high level of similarity in SDS-PAGE, fatty acid methyl esters and routine phenotypic tests. However, 16S rRNA gene sequence comparisons showed that this taxon was different from other related Bacillus species, and only a low level of DNA relatedness was found with the closest phylogenetic and phenotypic relative, Bacillus galactosidilyticus. This taxon could be differentiated from B. galactosidilyticus on the basis of morphological differences, stronger acid reactions with a wide range of substrates after 48 h incubation, and qualitative and quantitative differences in fatty acid content. On the basis of these data, a novel species, Bacillus ruris sp. nov., is proposed, with LMG 22866T (=DSM 17057T) as the type strain.


Candida tropicalis in Peritoneal Dialysis-related Peritonitis Diagnosed by Matrix-assisted Laser Desorption/ionization Time-of-flight Mass Spectrometry.
Fungal peritonitis (FP) is a rare complication of peritoneal dialysis (PD). Candida tropicalis infections are rarely reported in literature. The authors present the first case of FP with peritoneal abscess due to C. tropicalis in a 22-year-old woman admitted to our hospital with septic shock. Abdominal tomography demonstrated an abscess in peritoneal space and intraluminal bubble appearance in femoral venous and arterial circulation. PD fluid specimens and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis revealed fungus growth of C. tropicalis. Fluconazole therapy was administered accompanied by catheter removal and mechanical ventilation with vasopressor support. The patient recovered after 23 days of hospitalization and was discharged. FP represents high mobility and mortality unless infection source control and appropriate antimicrobial therapy are implemented accompanied by PD catheter removal. The use of MALDI-TOF MS for PD-related peritonitis pathogen identification can promote early identification and appropriate antibiotic therapy, especially in C. tropicalis infection.


Discovery and Structure-Based Optimization of Next-Generation Reversible Methionine Aminopeptidase-2 (MetAP-2) Inhibitors.
Co- and post-translational processing are crucial maturation steps to generate functional proteins. MetAP-2 plays an important role in this process, and inhibition of its proteolytic activity has been shown to be important for angiogenesis and tumor growth, suggesting that small-molecule inhibitors of MetAP-2 may be promising options for the treatment of cancer. This work describes the discovery and structure-based hit optimization of a novel MetAP-2 inhibitory scaffold. Of critical importance, a cyclic tartronic diamide coordinates the MetAP-2 metal ion in the active site while additional side chains of the molecule were designed to occupy the lipophilic methionine side chain recognition pocket as well as the shallow cavity at the opening of the active site. The racemic screening hit from HTS campaign 11a was discovered with an enzymatic IC50 of 150 nM. The resynthesized eutomer confirmed this activity and inhibited HUVEC proliferation with an IC50 of 1.9 μM. Its structural analysis revealed a sophisticated interaction pattern of polar and lipophilic contacts that were used to improve cellular potency to an IC50 of 15 nM. In parallel, the molecular properties were optimized on plasma exposure and antitumor efficacy which led to the identification of advanced lead 21.


Time-resolved neural reinstatement and pattern separation during memory decisions in human hippocampus.
Mnemonic decision-making has long been hypothesized to rely on hippocampal dynamics that bias memory processing toward the formation of new memories or the retrieval of old ones. Successful memory encoding may be best optimized by pattern separation, whereby two highly similar experiences can be represented by underlying neural populations in an orthogonal manner. By contrast, successful memory retrieval is thought to be supported by a recovery of the same neural pattern laid down during encoding. Here we examined how hippocampal pattern completion and separation emerge over time during memory decisions. We measured electrocorticography activity in the human hippocampus and posterior occipitotemporal cortex (OTC) while participants performed continuous recognition of items that were new, repeated (old), or highly similar to a prior item (similar). During retrieval decisions of old items, both regions exhibited significant reinstatement of multivariate high-frequency activity (HFA) associated with encoding. Further, the extent of reinstatement of encoding patterns during retrieval was correlated with the strength (HFA power) of hippocampal encoding. Evidence for encoding pattern reinstatement was also seen in OTC on trials requiring fine-grained discrimination of similar items. By contrast, hippocampal activity showed evidence for pattern separation during these trials. Together, these results underscore the critical role of the hippocampus in supporting both reinstatement of overlapping information and separation of similar events.


Usefulness of High-Density Lipoprotein Cholesterol to Predict Survival in Pulmonary Arterial Hypertension.
It has been suggested that lipoprotein abnormalities may contribute to the pulmonary arteriolar dysfunction observed in pulmonary arterial hypertension (PAH). High-density lipoprotein cholesterol (HDL) has vasodilatory, anti-inflammatory, and endothelial protective properties. We hypothesized that a higher serum HDL level may be advantageous for survival in PAH and that the serum HDL level at diagnosis would be an independent predictor of survival in PAH and be additive to previously validated predictors of survival. This study included all patients with PAH seen at the Mayo Clinic Pulmonary Hypertension Clinic from January 1, 1995, to December 31, 2009, who had a baseline HDL measurement. Mortality was analyzed over 5 years using the Kaplan-Meier method. Univariate and multivariable Cox proportional hazards ratios were calculated to evaluate the relation between baseline HDL level and survival. HDL levels were available for 227 patients. Higher HDL levels were associated with significantly lower mortality. Patients with an HDL >54 mg/dl at diagnosis had a 5-year survival of 59%. By comparison those with an HDL <34 mg/dl had a 5-year survival of 30%. On multivariate analysis, higher HDL was associated with an age-adjusted risk ratio for death of 0.78 (CI 0.67 to 0.91; p <0.01) per 10 mg/dl increase. In conclusion, HDL was an independent predictor of survival in PAH.


Conserved Nonexonic Elements: A Novel Class of Marker for Phylogenomics.
Noncoding markers have a particular appeal as tools for phylogenomic analysis because, at least in vertebrates, they appear less subject to strong variation in GC content among lineages. Thus far, ultraconserved elements (UCEs) and introns have been the most widely used noncoding markers. Here we analyze and study the evolutionary properties of a new type of noncoding marker, conserved nonexonic elements (CNEEs), which consists of noncoding elements that are estimated to evolve slower than the neutral rate across a set of species. Although they often include UCEs, CNEEs are distinct from UCEs because they are not ultraconserved, and, most importantly, the core region alone is analyzed, rather than both the core and its flanking regions. Using a data set of 16 birds plus an alligator outgroup, and ∼3600-∼3800 loci per marker type, we found that although CNEEs were less variable than bioinformatically derived UCEs or introns and in some cases exhibited a slower approach to branch resolution as determined by phylogenomic subsampling, the quality of CNEE alignments was superior to those of the other markers, with fewer gaps and missing species. Phylogenetic resolution using coalescent approaches was comparable among the three marker types, with most nodes being fully and congruently resolved. Comparison of phylogenetic results across the three marker types indicated that one branch, the sister group to the passerine + falcon clade, was resolved differently and with moderate (>70%) bootstrap support between CNEEs and UCEs or introns. Overall, CNEEs appear to be promising as phylogenomic markers, yielding phylogenetic resolution as high as for UCEs and introns but with fewer gaps, less ambiguity in alignments and with patterns of nucleotide substitution more consistent with the assumptions of commonly used methods of phylogenetic analysis.


A novel homozygous TPM1 mutation in familial pediatric hypertrophic cardiomyopathy and in silico screening of potential targeting drugs.
Familial hypertrophic cardiomyopathy (HCM) is the most common genetic cardiac disease. While sarcomeric gene mutations explain many HCM cases, the genetic basis of about half of HCM cases remains elusive. Here we aimed to identify the gene causing HCM in a non-consanguineous Saudi Arabian family with affected family members and a history of sudden death. The impact of the identified mutation on protein structure and potential drug targets were evaluated in silico. Triplets (two HCM subjects and one patent ductus arteriosus (PDA) case) and unaffected parents were screened by targeted next-generation sequencing (NGS) for 181 candidate cardiomyopathy genes. In silico structural and functional analyses, including protein modeling, structure prediction, drug screening, drug binding, and dynamic simulations were performed to explore the potential pathogenicity of the variant and to identify candidate drugs. A homozygous missense mutation in exon 1 of TMP1 (assembly GRCh37-chr15: 63340781; G>A) was identified in the triplets [two HCM and one patent ductus arteriosus (PDA)] that substituted glycine for arginine at codon 3 (p.Gly3Arg). The parents were heterozygous for the variant. The mutation was predicted to cause a significant and deleterious change in the TPM1 protein structure that slightly affected drug binding, stability, and conformation. In addition, we identified several putative TPM1-targeting drugs through structure-based in silico screening. TPM1 mutations are a common cause of HCM and other congenital heart defects. To date, TPM1 has not been associated with isolated PDA; to our knowledge, this is the first report of the homozygous missense variation p.Gly3Arg in TPM1 associated with familial autosomal recessive pediatric HCM and PDA. The identified candidate TPM1 inhibitors warrant further prospective investigation.


Genes Encoding Transcription Factors TaDREB5 and TaNFYC-A7 Are Differentially Expressed in Leaves of Bread Wheat in Response to Drought, Dehydration and ABA.
Two groups of six spring bread wheat varieties with either high or low grain yield under the dry conditions of Central and Northern Kazakhstan were selected for analysis. Experiments were set up with the selected wheat varieties in controlled environments as follows: (1) slowly progressing drought imposed on plants in soil, (2) rapid dehydration of whole plants grown in hydroponics, (3) dehydration of detached leaves, and (4) ABA treatment of whole plants grown in hydroponics. Representatives of two different families of transcription factors (TFs), TaDREB5 and TaNFYC-A7, were found to be linked to yield-under-drought using polymorphic Amplifluor-like SNP marker assays. qRT-PCR revealed differing patterns of expression of these genes in the leaves of plants subjected to the above treatments. Under drought, TaDREB5 was significantly up-regulated in leaves of all high-yielding varieties tested and down-regulated in all low-yielding varieties, and the level of expression was independent of treatment type. In contrast, TaNFYC-A7 expression levels showed different responses in the high- and low-yield groups of wheat varieties. TaNFYC-A7 expression under dehydration (treatments 2 and 3) was higher than under drought (treatment 1) in all high-yielding varieties tested, while in all low-yielding varieties the opposite pattern was observed: the expression levels of this gene under drought were higher than under dehydration. Rapid dehydration of detached leaves and intact wheat plants grown in hydroponics produced similar changes in gene expression. ABA treatment of whole plants caused rapid stomatal closure and a rise in the transcript level of both genes during the first 30 min, which decreased 6 h after treatment. At this time-point, expression of TaNFYC-A7 was again significantly up-regulated compared to untreated controls, while TaDREB5 returned to its initial level of expression. These findings reveal significant differences in the transcriptional regulation of two drought-responsive and ABA-dependent TFs under slowly developing drought and rapid dehydration of wheat plants. The results obtained suggest that correlation between grain yield in dry conditions and TaNFYC-A7 expression levels in the examined wheat varieties is dependent on the length of drought development and/or strength of drought; while in the case of TaDREB5, no such dependence is observed.


Fabrication of controlled-release silver nanoparticle polylactic acid microneedles with long-lasting antibacterial activity using a micro-molding solvent-casting technique.
Most topical drug delivery techniques do not provide therapeutic concentrations for treatment of surgical site and other local infections and, therefore, require some kind of enhancement, such as physical methods like microneedles, the subject of the present investigation. Here, controlled-release long-lasting antibacterial polylactic acid (PLA) microneedles containing 1, 3, and 5% silver nanoparticles (AgNP) were prepared using micro-molding solvent-casting technique. Microneedles were characterized using optical microscopy, SEM, FTIR, XRD, and DSC. Also, mechanical strength, barrier disruption ability, insertion depth, in-vitro release kinetics, antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa, and silver permeation through rat skin were studied. Microneedles showed good mechanical strength with no signs of failure at an optimum PLA concentration of 25% (w/v). FTIR revealed no chemical interaction between ingredients, and XRD confirmed presence of AgNP in microneedles. Microneedles penetrated the skin model at depth of up to 1143 μm resulting 5-7 times increase in transepidermal water loss (TEWL). Release studies showed 2.2, 6.8, and 8.1 µg silver release from the whole body (obeying Higuchi's release model) and 0.33, 0.45, and 0.78 µg from the needles alone (obeying Fickian-cylindrical type release) for 1, 3, and 5% AgNP microneedles, respectively. Also, prolonged antibacterial activity (for 34 days) was observed. Skin studies over 72 h indicated that besides needles, silver is also released from the baseplate which had a marginal share in total silver permeation through the skin. In conclusion, a straightforward solvent-casting technique can be used to successfully prepare strong AgNP-containing PLA microneedles capable of long-lasting antibacterial activity.


Palm tree syrup: nutritional composition of a natural edulcorant.
Palm syrup is a typical product from the Canary Islands, traditionally produced from the sap of the tropical palm tree Phoenix canariensis. Its high caloric content has led to its increasing use as a health food supplement for athletes, children and elderly. Furthermore, demand for this natural syrup is continuously increasing due also to its medicinal uses in homeopathic medicine. Palm Tree syrup samples prepared with palm sap from primary producers in La Gomera island (Canary Islands, Spain) were analyzed for their nutritional composition (moisture, ash, sugars, fat, vitamins and minerals). 35 syrup samples from five different producing regions in La Gomera island were analyzed. High-performance liquid chromatography (HPLC) was used to determine sugars and vitamins and Flame Atomic Absorption Spectrophotometry (FAAS) was used to analyze the minerals. Major carbohydrates were sucrose (37.8%), glucose (9.50%) and fructose (4.80%), respectively. The presence of arabinose could not be confirmed. Niacin was the water-soluble vitamin with the highest concentration with an average content of 0.003%. Fat content was found to be under 0.20%. Potassium was the mineral with highest contents (0.45%). Results suggest that palm tree syrup can play an important role as a sugar and mineral source in human nutrition, suggesting that future applications for this product could be developed.


Estimation of Aboveground Biomass in Agroforestry Systems over Three Climatic Regions in West Africa Using Sentinel-1, Sentinel-2, ALOS, and GEDI Data.
Agroforestry systems (AFS) offer viable solutions for climate change because of the aboveground biomass (AGB) that is maintained by the tree component. Therefore, spatially explicit estimation of their AGB is crucial for reporting emission reduction efforts, which can be enabled using remote sensing (RS) data and methods. However, multiple factors including the spatial distributions within the AFS, their structure, their composition, and their variable extents hinder an accurate RS-assisted estimation of the AGB across AFS. The aim of this study is to (i) evaluate the potential of spaceborne optical, SAR and LiDAR data for AGB estimations in AFS and (ii) estimate the AGB of different AFS in various climatic regions. The study was carried out in three climatic regions covering Côte d'Ivoire and Burkina Faso. Two AGB reference data sources were assessed: (i) AGB estimations derived from field measurements using allometric equations and (ii) AGB predictions from the GEDI level 4A (L4A) product. Vegetation indices and texture parameters were generated from optical (Sentinel-2) and SAR data (Sentinel-1 and ALOS-2) respectively and were used as predictors. Machine learning regression models were trained and evaluated by means of the coefficient of determination (R2) and the RMSE. It was found that the prediction error was reduced by 31.2% after the stratification based on the climatic conditions. For the AGB prediction, the combination of random forest algorithm and Sentinel-1 and -2 data returned the best score. The GEDI L4A product was applicable only in the Guineo-Congolian region, but the prediction error was approx. nine times higher than the ground truth. Moreover, the AGB level varied across AFS including cocoa (7.51 ± 0.6 Mg ha-1) and rubber (7.33 ± 0.33 Mg ha-1) in the Guineo-Congolian region, cashew (13.78 ± 0.98 Mg ha-1) and mango (12.82 ± 0.65 Mg ha-1) in the Guinean region. The AFS farms in the Sudanian region showed the highest AGB level (6.59 to 82.11 Mg ha-1). AGB in an AFS was mainly determined by the diameter (R2 = 0.45), the height (R2 = 0.13) and the tree density (R2 = 0.10). Nevertheless, RS-based estimation of AGB remain challenging because of the spectral similarities between AFS. Therefore, spatial assessment of the prediction uncertainties should complement AGB maps in AFS.


betaDelta and betaSandwich: Confidence Intervals for Standardized Regression Coefficients in R.
The multivariate delta method was used by Yuan and Chan to estimate standard errors and confidence intervals for standardized regression coefficients. Jones and Waller extended the earlier work to situations where data are nonnormal by utilizing Browne's asymptotic distribution-free (ADF) theory. Furthermore, Dudgeon developed standard errors and confidence intervals, employing heteroskedasticity-consistent (HC) estimators, that are robust to nonnormality with better performance in smaller sample sizes compared to Jones and Waller's ADF technique. Despite these advancements, empirical research has been slow to adopt these methodologies. This can be a result of the dearth of user-friendly software programs to put these techniques to use. We present the betaDelta and the betaSandwich packages in the R statistical software environment in this manuscript. Both the normal-theory approach and the ADF approach put forth by Yuan and Chan and Jones and Waller are implemented by the betaDelta package. The HC approach proposed by Dudgeon is implemented by the betaSandwich package. The use of the packages is demonstrated with an empirical example. We think the packages will enable applied researchers to accurately assess the sampling variability of standardized regression coefficients.


A Glycine soja group S2 bZIP transcription factor GsbZIP67 conferred bicarbonate alkaline tolerance in Medicago sativa.
Even though bicarbonate alkaline stress is a serious threat to crop growth and yields, it attracts much fewer researches than high salinity stress. The basic leucine zipper (bZIP) transcription factors have been well demonstrated to function in diverse abiotic stresses; however, their biological role in alkaline tolerance still remains elusive. In this study, we functionally characterized a bZIP gene from Glycine soja GsbZIP67 in bicarbonate alkaline stress responses. GsbZIP67 was initially identified as a putative bicarbonate responsive gene, on the basis of previous RNA-seq data of 50 mM NaHCO3-treated Glycine soja roots. GsbZIP67 protein possessed a conserved bZIP domain, and belonged to the group S2 bZIP, which is yet less well-studied. Our studies showed that GsbZIP67 targeted to nucleus in Arabidopsis protoplasts, and displayed transcriptional activation activity in yeast cells. The quantitative real-time PCR analyses unraveled the bicarbonate stress responsive expression and tissue specific expression of GsbZIP67 in wild soybean. Further phenotypic analysis illustrated that GsbZIP67 overexpression in alfalfa promoted plant growth under bicarbonate alkaline stress, as evidenced by longer roots and shoots. Furthermore, GsbZIP67 overexpression also modified the physiological indices of transgenic alfalfa under bicarbonate alkaline stress. In addition, the expression levels of several stress responsive genes were also augmented by GsbZIP67 overexpression. Collectively, in this study, we demonstrated that GsbZIP67 acted as a positive regulator of plant tolerance to bicarbonate alkaline stress. These results provide direct genetic evidence of group S2 bZIPs in bicarbonate alkaline stress, and will facilitate further studies concerning the cis-elements and/or downstream genes targeted by GsbZIP67 in stress responses.


Use, costs, and predictors of psychiatric healthcare services following an autism spectrum diagnosis: Population-based cohort study.
A number of cross-sectional studies report extensive use of psychiatric services and high healthcare costs in autistic youths. However, little is known about how the use of these services evolves from the time of diagnosis, as children grow up. Our objectives were to investigate the use, costs, and predictors of psychiatric services following autism spectrum diagnosis. We built a cohort of 1227 newly diagnosed autism spectrum individuals identified in the Quebec (Canada) Régie de l'assurance maladie du Québec administrative database (January 1998 to December 2010). Mean number and cost per individual of psychiatric healthcare use (hospitalizations, medical visits, psychoactive drug use) were calculated yearly for 5 years following autism spectrum diagnosis. Mean number of psychiatric visits decreased over time by more than threefold (7.5 vs 2.1 visits) from year 1 to year 5, whereas psychoactive drug use increased from 16.0 to 25.2 claims. Psychiatric hospitalizations decreased during follow-up, but still represented the greatest costs per individual (CAD9820 for year 1; CAD4628 for year 5). Antipsychotics represented over 50% of drug costs. Mixed-effect model with repeated measures showed that previous psychoactive drug use was the strongest predictor of greater psychiatric healthcare cost during follow-up (odds ratio: 9.96; 95% confidence interval: 7.58-13.10). These trends contrast with guidelines advocating cautious prescribing of antipsychotics with periodical re-assessment of their benefit.


Alcohol and trauma: the critical link.
The link between traumatic injury and alcohol consumption is strong. Although statistics vary, from one half to one third of trauma victims admitted to critical care units have an alcohol-related injury. The initial role of the critical care nurse is to identify the presence of alcohol by monitoring the patient's BAC. Once the presence of alcohol is confirmed, its effects impact directly on the plan of care that evolves during the critical illness. Nursing interventions focus on four specific areas affected by alcohol: the ability of alcohol to mask injury, the effects of alcohol on medications, alcohol withdrawal, and rehabilitation from alcoholism. By incorporating these four areas into the plan of care, the critical care nurse assists the patient not only to recover from a traumatic injury but also to deal with a problem drinking pattern.


Are children and adolescents with psychiatric illness at risk for increased future body weight? A systematic review.
The aim of this paper was to review the longitudinal relationship between early-onset depression and disruptive behaviour and adult body weight. A systematic review of prospective longitudinal studies was conducted of articles in which (1) initial assessment occurred during childhood or adolescence (<18 y); and (2) the primary outcome reported as body mass index (BMI), overweight (BMI>25-<30 kg/m(2)), obesity (BMI ≥ 30 kg/m(2)), or depression; and (3) validated assessment measures for assessment of depressive symptoms or disruptive behaviour problems were employed. A total number of 16 articles were identified for review. Obese adolescent females are more likely to develop depressive illness in adulthood than their non-obese peers. Conversely, depressed adolescent females, and possibly males, are more likely to become overweight adults than non-depressed adolescents. There are insufficient data addressing future depression risk among overweight, non-obese, adolescents to evaluate the potentially interactive nature of this relationship. Studies to date are consistent in reporting that children with behaviour problems are at increased risk of future overweight, though whether this risk is conferred by conduct symptoms, ADHD symptoms, or both, is less clear. Care providers of children with disruptive behaviour problems and depressed adolescents should monitor weight gain. Among obese adolescent females, mood should be followed.


Decrease in Population and Increase in Welfare of Community Cats in a Twenty-Three Year Trap-Neuter-Return Program in Key Largo, FL: The ORCAT Program.
The objective of this study was to evaluate the effect of a long-term (23-year) trap-neuter-return program on the population size of community cats in the Ocean Reef Community and to describe the demographic composition and outcome of enrolled cats. A retrospective study was performed using both cat census data collected between 1999 and 2013 as well as individual medical records for cats whose first visit occurred between 3/31/1995 and 12/31/2017. Medical record entries were reviewed to determine program inputs, cat outcomes, retroviral disease prevalence, and average age of first visit, sterilization, and death through 6/11/2018. Change over time was analyzed via linear regression. The free-roaming population decreased from 455 cats recorded in 1999 to 206 recorded in 2013 (55% decrease, P < 0.0001). There were 3,487 visits recorded for 2,529 community cats, with 869 ovariohysterectomies and 822 orchiectomies performed. At last recorded visit, there were 1,111 cats returned back to their original location, and 1,419 cats removed via adoption (510), transfer to the adoption center (201), euthanasia of unhealthy or retrovirus positive cats (441), died in care (58), or outcome of dead on arrival (209). The number of first visits per year decreased 80% from 348 in 1995 to 68 in 2017. The estimated average age of the active cat population increased by 0.003 months each year (P = 0.031) from 16.6 months in 1995 to 43.8 months in 2017. The mean age of cats at removal increased 1.9 months per year over time (P < 0.0001) from 6.4 months in 1995 to 77.3 months in 2017. The mean age of cats at return to the original location was 20.8 months, which did not change over time. The overall retrovirus prevalence over the entire duration was 6.5%, with FIV identified in 3.3% of cats and FeLV identified in 3.6%. Retrovirus prevalence decreased by 0.32% per year (P = 0.001), with FIV decreasing by 0.16% per year (P = 0.013) and FeLV decreasing 0.18% per year (P = 0.033). In conclusion, a trap-neuter-return program operating for over two decades achieved a decrease in population and an increase in population welfare as measured by increased average age of population and decreased retrovirus prevalence.


Prophylactic use of gentamicin in colonic and rectal surgery.
A double-blind trial was performed to test whether a single intramuscular injection of 80 mg of gentamicin would significantly reduce the incidence of septic complications of elective colonic and rectal surgery. The gentamicin was given at the time of anaesthetic premedication, the operations were performed by either of two surgeons, and the patients were followed up by an independent investigator. Eighty patients satisfactorily completed the trial. There was no significant difference in the incidence of postoperative wound infection, intraperitoneal abscess formation and faecal fistula between the 41 patients who received the gentamicin and the 39 who received the placebo. However, the mortality was significantly lower in the gentamicin group, in which there was only one death.


Effect of synthetic peptides representing the hypervariable region of p21ras on Xenopus laevis oocyte maturation.
The carboxy-terminal hypervariable regions of p21ras proteins have been highly conserved throughout evolution but no function has been assigned to them yet. This region has been suggested as a possible candidate for receptor recognition. We have tested the possibility of this region being involved in p21ras biological function. Synthetic peptides corresponding to the hypervariable domains of p21N-ras and p21K(B)-ras were microinjected into Xenopus oocytes to assess their effect on oocyte maturation. The K(B)-ras peptide inhibited insulin-dependent but not progesterone-dependent maturation, in contrast with the N-ras peptide which did not inhibit maturation significantly. A control peptide, with the same amino acid composition as the K(B)-ras peptide but with a scrambled sequence, and poly(D,L-lysine) were inactive. Pentalysine had partial activity which may be due to its mimicking the lysine-rich stretch of the K(B)-ras sequence. The data support the hypothesis that the K(B)-ras gene product specifically is involved in transducing the insulin and/or insulin-like growth factor 1 signal.


Construction and validation of anisotropic and orthotropic ventricular geometries for quantitative predictive cardiac electrophysiology.
Reaction-diffusion computational models of cardiac electrophysiology require both dynamic excitation models that reconstruct the action potentials of myocytes as well as datasets of cardiac geometry and architecture that provide the electrical diffusion tensor D, which determines how excitation spreads through the tissue. We illustrate an experimental pipeline we have developed in our laboratories for constructing and validating such datasets. The tensor D changes with location in the myocardium, and is determined by tissue architecture. Diffusion tensor magnetic resonance imaging (DT-MRI) provides three eigenvectors e(i) and eigenvalues λ(i) at each voxel throughout the tissue that can be used to reconstruct this architecture. The primary eigenvector e(1) is a histologically validated measure of myocyte orientation (responsible for anisotropic propagation). The secondary and tertiary eigenvectors (e(2) and e(3)) specify the directions of any orthotropic structure if λ(2) is significantly greater than λ(3)-this orthotropy has been identified with sheets or cleavage planes. For simulations, the components of D are scaled in the fibre and cross-fibre directions for anisotropic simulations (or fibre, sheet and sheet normal directions for orthotropic tissues) so that simulated conduction velocities match values from optical imaging or plunge electrode experiments. The simulated pattern of propagation of action potentials in the models is partially validated by optical recordings of spatio-temporal activity on the surfaces of hearts. We also describe several techniques that enhance components of the pipeline, or that allow the pipeline to be applied to different areas of research: Q ball imaging provides evidence for multi-modal orientation distributions within a fraction of voxels, infarcts can be identified by changes in the anisotropic structure-irregularity in myocyte orientation and a decrease in fractional anisotropy, clinical imaging provides human ventricular geometry and can identify ischaemic and infarcted regions, and simulations in human geometries examine the roles of anisotropic and orthotropic architecture in the initiation of arrhythmias.


Aharonov-Bohm interferences in polycrystalline graphene.
Aharonov-Bohm (AB) interferences in the quantum Hall regime can be achieved, provided that electrons are able to transmit between two edge channels in nanostructures. Pioneering approaches include quantum point contacts in 2DEG systems, bipolar graphene p-n junctions, and magnetic field heterostructures. In this work, defect scattering is proposed as an alternative mechanism to achieve AB interferences in polycrystalline graphene. Indeed, due to such scattering, the extended defects across the sample can act as tunneling paths connecting quantum Hall edge channels. Consequently, strong AB oscillations in the conductance are predicted in polycrystalline graphene systems with two parallel grain boundaries. In addition, this general approach is demonstrated to be applicable to nano-systems containing two graphene barriers with functional impurities and perspectively, can also be extended to similar systems of 2D materials beyond graphene.


The development of template-based facial expression perception from 6 to 15 years of age.
When perceiving emotional facial expressions, adults use a template-matching strategy, comparing the perceived face with a stored representation. A rejection of unnaturally exaggerated faces is characteristic of this strategy because the exaggerated expressions do not match the stored template. In contrast, a rule-based perceptual strategy (e.g., wide eyes indicate surprise) would be more tolerant of exaggeration. The current study uses exaggeration tolerance to test the expression perception strategies of children from 6 to 15 years of age. In Experiment 1, 62 (38 male) participants viewed pairs of happy or sad faces varying in exaggeration and selected the face that looked closest to how a happy (or sad) person really looks. With age, children became less likely to choose the more exaggerated expression. In Experiment 2, this result was replicated with each of the six basic emotions. Sixty-six children (26 male, 50 Caucasian, 10 mixed-race, four Indian, two unidentified) from 6 to 15 years of age completed the same experimental tasks as Experiment 1 for all six emotions. Again, with age children became less likely to choose the more exaggerated face. The results from both experiments suggest that the development of an adult-like template-matching strategy lasts into adolescence. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


[Prognostic significance of flow cytophotometric studies in ovarian cancer].
Prognostic significance of DNA content and of distribution of cell cycle phases in ovarian carcinomas (stage III and IV) were investigated using flow cytometry. From 37 tumors 15 had DNA indices less than 2.5 and 22 had DNA indices greater than or equal to 2.5 (diploid = 2). Patients with diploid or near diploid tumors had significantly longer survival times than those with aneuploid tumors (DNA indices greater than 2.5) (log-rank test, p = 0.009, rank-sum test, p = 0.011). Patients whose tumors showed a high proportion of S/G2/M-phase cells (greater than 17%) had shorter survival times than those with tumors with a lower proportion of S/G2/M-phase cells (log-rank test p = 0.009, rank-sum test p = 0.022). The median length of follow-up of the patients was 4 years. Thus, our experiments have shown that measurements of DNA ploidy and proliferative activity using flow cytometry are important prognostic indicators for patients with ovarian carcinomas. In the future these factors might play an important role for the prognosis of patients with ovarian carcinomas. In addition, there exists a tendency for patients with in vitro resistant tumors to die earlier than those with sensitive tumors, but these results were not significant (log-rank test, p = 0.16; rank-sum test, p = 0.059).


HISTOCHEMICAL DEMONSTRATION OF UPTAKE OF EXOGENOUS NOREPINEPHRINE BY ADRENERGIC FIBERS IN VITRO.
Adrenergic fibers in isolated iris preparations were demonstrated histochemically by means of the formaldehyde condensation technique. In the presence of adenosine triphosphate (ATP) there was a net uptake of norepinephrine added in vitro by the adrenergic fibers producing a striking increase in fluorescence. In the absence of ATP only a slight increase in fluorescence was observed when the tissue was exposed to high concentrations of norepinephrine.


Click and Detect: Versatile Ampicillin Aptasensor Enabled by Click Chemistry on a Graphene-Alkyne Derivative.
Tackling the current problem of antimicrobial resistance (AMR) requires fast, inexpensive, and effective methods for controlling and detecting antibiotics in diverse samples at the point of interest. Cost-effective, disposable, point-of-care electrochemical biosensors are a particularly attractive option. However, there is a need for conductive and versatile carbon-based materials and inks that enable effective bioconjugation under mild conditions for the development of robust, sensitive, and selective devices. This work describes a simple and fast methodology to construct an aptasensor based on a novel graphene derivative equipped with alkyne groups prepared via fluorographene chemistry. Using click chemistry, an aptamer is immobilized and used as a successful platform for the selective determination of ampicillin in real samples in the presence of interfering molecules. The electrochemical aptasensor displayed a detection limit of 1.36 nM, high selectivity among other antibiotics, the storage stability of 4 weeks, and is effective in real samples. Additionally, structural and docking simulations of the aptamer shed light on the ampicillin binding mechanism. The versatility of this platform opens up wide possibilities for constructing a new class of aptasensor based on disposable screen-printed carbon electrodes usable in point-of-care devices.


Transcriptional mapping of the varicella-zoster virus regulatory genes encoding open reading frames 4 and 63.
Four of the 68 varicella-zoster virus (VZV) unique open reading frames (ORFs), i.e., ORFs 4, 61, 62, and 63, encode proteins that influence viral transcription and are considered to be positional homologs of herpes simplex virus type 1 (HSV-1) immediate-early (IE) proteins. In order to identify the elements that regulate transcription of VZV ORFs 4 and 63, the encoded mRNAs were mapped in detail. For ORF 4, a major 1.8-kb and a minor 3.0-kb polyadenylated [poly(A)+] RNA were identified, whereas ORF 63-specific probes recognized 1.3- and 1.9-kb poly(A)+ RNAs. Probes specific for sequences adjacent to the ORFs and mapping of the RNA 3' ends indicated that the ORF 4 RNAs were 3' coterminal, whereas the RNAs for ORF 63 represented two different termination sites. S1 nuclease mapping and primer extension analyses indicated a single transcription initiation site for ORF 4 at 38 bp upstream of the ORF start codon. For ORF 63, multiple transcriptional start sites at 87 to 95, 151 to 153, and (tentatively) 238 to 243 bp upstream of the ORF start codon were identified. TATA box motifs at good positional locations were found upstream of all mapped transcription initiation sites. However, no sequences resembling the TAATGARAT motif, which confers IE regulation upon HSV-1 IE genes, were found. The finding of the absence of this motif was supported through analyses of the regulatory sequences of ORFs 4 and 63 in transient transfection assays alongside those of ORFs 61 and 62. Sequences representing the promoters for ORFs 4, 61, and 63 were all stimulated by VZV infection but failed to be stimulated by coexpression with the HSV-1 transactivator Vmw65. In contrast, the promoter for ORF 62, which contains TAATGARAT motifs, was activated by VZV infection and coexpression with Vmw65. These results extend the transcriptional knowledge for VZV and suggest that ORFs 4 and 63 contain regulatory signals different from those of the ORF 62 and HSV-1 IE genes.


UPLC-ESI/MS determination of 17 active constituents in two categorized formulas of traditional Chinese medicine, Sanhuang Xiexin Tang and Fuzi Xiexin Tang: application in comparing the differences in decoctions and macerations.
A rapid and sensitive UPLC-ESI/MS method was established and validated to determine 17 active constituents (aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, berberine, palmatine, jatrorrhizine, coptisine, baicalein, wogonin, baicalin, wogonoside, emodin, aloe-emodin and rhein) in Sanhuang Xiexin Tang (SXT) and Fuzi Xiexin Tang (FXT), which are two classic compound recipes from Xiexin Tang categorized formulas in traditional Chinese medicien. The separation was performed on a UPLC BEH C18 column gradient eluted using acetonitrile and 0.1% formic acid as mobile phase. ESI/MS was operated in positive ([M + H](+)) in selected ion recording mode for analysis of alkaloids and flavones, while in negative ([M - H](-)) selected ion recording mode for anthraquinones. All of the 17 constituents exhibited good linearity in a relatively wide concentration ranges with the lowest limits of detection of 0.38 ng/mL. All of the relative standard deviation values of intra- and inter-precisions and stabilities of 17 constituents were within 5%. The method was successfully applied to determine 17 active constituents in decoctions and macerations of SXT and FXT. The results indicated that different preparative methods resulted in significant diversity in concentrations of the 17 analytes. Herb-herb interaction appeared between aconitum alkaloids in Aconiti Lateralis Radix Preparata and another three herbs.


Orexins increase penicillin-induced epileptic activity.
Orexins have been implicated with physiological function including sleep-wake cycle, energy homeostasis, drinking behavior, analgesia, attention, learning and memory but their effects on excitability are controversial. We investigated the effects of intracortical injections of orexin A (100 pmol) and B (100 pmol) on the electrophysiological manifestation of epileptic seizures induced by cortical penicillin application in adult male rats. In comparison to saline, orexin A and B enhanced significantly the spike number, spike amplitude and spectral power values induced by cortical penicillin. Our findings indicates that orexins enhances the hyperexcitable and hypersyncronic cortical epileptic activity induced by focal application of penicillin-G.


Injury risk in professional boxing.
Although a popular endeavor, boxing has fallen under increased scrutiny because of its association with traumatic brain injury. However, few studies have investigated the overall epidemiology of boxing injuries from representative samples, and no study has ever documented the incidence of injuries in female boxers. This study is a review of professional boxing data from the state of Nevada from September 2001 through March 2003. Medical and outcome data for all professional boxing matches occurring in Nevada between September 2001 and March 2003 (n = 524 matches) were analyzed on the basis of a pair-matched, case-control design. Cases were boxers who received an injury during the boxing matches. Boxers who were not injured served as control subjects. Both conditional and unconditional logistic regression models were used to assess risk factors for injury. The overall incidence rate of injury was 17.1 per 100 boxer-matches, or 3.4 per 100 boxer-rounds. Facial laceration accounted for 51% of all injuries, followed by hand injury (17%), eye injury (14%), and nose injury (5%). Male boxers were significantly more likely than female boxers to receive injuries (3.6 versus 1.2 per 100 boxer-rounds, P = 0.01). Male boxing matches also ended in knockouts and technical knockouts more often than did female matches (P < 0.001). The risk of injury for those who lost the matches was nearly twice the risk for the winners. Those who lost by knockout had double the risk of injury compared with those who lost by other means. Neither age nor weight was significantly associated with the risk of injury. The injury rate in professional boxing matches is high, particularly among male boxers. Superficial facial lacerations are the most common injury reported. Male boxers have a higher rate of knockout and technical knockouts than female boxers. Further research is necessary to determine the outcomes of injury, particularly the long-term neurologic outcome differences between sexes.


Induction of TLR tolerance in human macrophages by adiponectin: does LPS play a role?
Obesity is regarded as a pro-inflammatory state. It is associated with low circulating levels of the adipokine, adiponectin, which is considered to be an anti-inflammatory. However, adiponectin knockout mice do not consistently demonstrate pro-inflammatory phenotypes, suggesting more complexity in the in vivo immunomodulatory effects of adiponectin than originally anticipated. Moreover, adiponectin exerts pro-inflammatory effects in some experimental systems. This contradiction has been resolved by hypothesizing that adiponectin induces tolerance to inflammatory stimuli, notably Toll-like receptor (TLR) ligands. We noticed that this effect resembled lipopolysaccharide (LPS) tolerance and therefore tested adiponectin from a variety of sources for LPS contamination. All adiponectin tested carried low levels of LPS in the range of 1-30 pg/microg of adiponectin, sufficient to produce final LPS concentrations in the pg/ml range under experimental conditions. We found that induction of tolerance to TLR ligands by adiponectin in human monocyte-derived macrophages could be reproduced by such LPS concentrations. Moreover, the LPS antagonist, polymixin B, substantially inhibited induction of tolerance by adiponectin. Furthermore, polymixin B and a naturally occurring antagonist LPS were able to partially attenuate induction of tumour necrosis factor-alpha and interleukin-6 in human monocyte-derived macrophages by adiponectin. Polymixin B also inhibited nuclear factor-kappaB and mitogen-activated protein kinase signalling elicited by adiponectin. We therefore propose that some of adiponectin's immunomodulatory effects, in particular, its TLR-tolerising actions in human monocyte-derived macrophages, may be confounded by induction of tolerance by contaminating LPS.


Role of Quantitative Computed Tomographic Scan Analysis in Lung Volume Reduction for Emphysema.
Recent advances in bronchoscopic lung volume reduction (BLVR) offer new therapeutic alternatives for patients with emphysema and hyperinflation. Endobronchial valves and coils are 2 potential BLVR techniques which have been shown to improve pulmonary function and the quality of life in patients with emphysema. Current patient selection for LVR procedures relies on 3 main inclusion criteria: low attenuation area (in %), also known as emphysema score, heterogeneity score, and fissure integrity score. Volumetric analysis in combination with densitometric analysis of the affected lung lobe or segment with quantitative CT to determine emphysema severity play an important role in treatment planning and post-operative assessment. Due to the variations in lung anatomy, manual corrections are often required to ensure successful and accurate lobe segmentation for pathological and post-treatment CT scan analysis. The advanced development and utilisation of quantitative CT do not simply represent regional changes in pulmonary function but aids in analysis for better patient selection with severe emphysema who are most likely to benefit from BLVR.


Super-resolution imaging detects BP180 autoantigen in immunoglobulin M pemphigoid.
Bullous pemphigoid is generally caused by immunoglobulin (Ig)G autoantibodies against hemidesmosomal BP180 and/or BP230. Recently, the concept of IgM pemphigoid has been proposed. A 23-year-old Japanese woman presented with a 4-month history of severely itchy papules showing subepidermal separations with mild neutrophil infiltration. Direct immunofluorescence (DIF) revealed IgM deposits at the dermoepidermal junction, but neither IgG nor IgA deposits. Indirect immunofluorescence on 1 M NaCl-split skin demonstrated deposits on the epidermal side. The optical density (OD) value of a modified IgM enzyme-linked immunosorbent assay for full-length BP180, but not for BP180-NC16A, was increased. The patient was diagnosed with IgM pemphigoid and was treated with diphenyl sulfone at 50 mg/day without recurrence. To confirm the precise autoantigen, we tried to obtain super-resolution imaging. The deposition pattern of IgM autoantibodies seemed to be oriented parallel to that of BP180. The detailed images detect DIF deposits apart from BP180-NC16A staining, but are close to type VII collagen-NC1 staining. This result suggests that the IgM autoantibodies in the patient might target the C-terminus of BP180. IgM pemphigoid is still not a widely accepted concept, and the clinical course remains unknown. We will carefully follow-up the patient. Super-resolution images may help to detect precise autoantigens in autoimmune blistering diseases.


Effect of electroacupuncture at Ximen (PC 4) and Hegu (LI 4) on expression of Akt in rats with myocardial ischemia-reperfusion injury.
To investigate the effect of electroacupuncture (EA) at acupoints on the pericardium meridian on the expression of phosphorylated Akt (p-Akt) protein in rat myocardium after ischemia and reperfusion. Seventy Wistar rats were evenly randomized into seven groups: the sham operation group (group A), ischemia-reperfusion model Ⅰ group (group B), ischemia-reperfusion model Ⅱ group (group C), EA at Neiguan (PC 6) group (group D), EA at Ximen (PC 4) group (group E), EA at Hegu (LI 4) group (group F), and LY294002 + EA at Neiguan (PC 6) group (group G). All processes were monitored by electrocardiography. In group A, the left anterior descending coronary artery was only threaded without ligation for 100 min. In group B, the left anterior descending coronary artery was ligated for 40 min and reperfused for 60 min. The left anterior descending coronary artery in group C was ligated for 40 min and reperfused for 100 min. Groups D, E, and F received EA for 20 min before undergoing ischemia for 40 min, and then received EA for 20 min before undergoing reperfusion for 60 min. Before modeling, group G was injected with LY294002 (0.3 mg/kg) into the tail vein, and then underwent the same intervention as the other EA groups. After reperfusion, myocardial tissue from the left cardiac ventricle was collected to enable Western blot analysis of the p-Akt level, and analysis of electrocardiographic changes. In groups B and C, electrocardiography showed obvious elevation of the ST-segment Ⅱ lead (ECG-STⅡ), while the ECG-ST Ⅱ values were significantly lower in groups D, E, and G (P < 0.01). The p-Akt levels in groups D and E were significantly greater than those in groups B and C (P < 0.01). Compared with all other groups, group G showed a significantly different expression of p-Akt (P < 0.01). The expression of p-Akt protein in cardiomyocytes was significantly greater in rats that were injected with LY294002 and received EA at Ximen (PC 4) compared with all other groups. This suggests that EA at Ximen (PC 4) resulted in activation of the phosphoinositide 3-kinase/Akt signaling pathway and phosphorylation of Akt.


Diabetic nephropathy: early detection, prevention and management.
Diabetic nephropathy typically presents more than a decade after diagnosis of diabetes and correlates with the duration of poorly controlled disease. Diabetic nephropathy begins as glomerular hypertension and hyperfiltration, followed by microalbuminuria and the development of hypertension, overt proteinuria, nephrotic syndrome, and a progressive decline in the glomerular filtration rate. Increasing expansion of the glomerular mesangium correlates with loss of function, resulting in uremia. This process eventually leads to the need for dialysis or renal transplantation in 30 percent of patients with insulin-dependent diabetes. By lowering intraglomerular pressure through enhanced glycemic control, inhibition of angiotensin and limitation of protein intake, severe nephropathy may be prevented, delayed or even partially reversed. Treatment must stress control of hypertension.


Vacuum-assisted headspace solid phase microextraction of polycyclic aromatic hydrocarbons in solid samples.
For the first time, Vacuum Assisted Headspace Solid Phase Microextraction (Vac-HSSPME) is used for the recovery of polycyclic aromatic hydrocarbons (PAHs) from solid matrices. The procedure was investigated both theoretically and experimentally. According to the theory, reducing the total pressure increases the vapor flux of chemicals at the soil surface, and hence improves HSSPME extraction kinetics. Vac-HSSPME sampling could be further enhanced by adding water as a modifier and creating a slurry mixture. For these soil-water mixtures, reduced pressure conditions may increase the volatilization rates of compounds with a low K(H) present in the aqueous phase of the slurry mixture and result in a faster HSSPME extraction process. Nevertheless, analyte desorption from soil to water may become a rate-limiting step when significant depletion of the aqueous analyte concentration takes place during Vac-HSSPME. Sand samples spiked with PAHs were used as simple solid matrices and the effect of different experimental parameters was investigated (extraction temperature, modifiers and extraction time). Vac-HSSPME sampling of dry spiked sand samples provided the first experimental evidence of the positive combined effect of reduced pressure and temperature on HSSPME. Although adding 2 mL of water as a modifier improved Vac-HSSPME, humidity decreased the amount of naphthalene extracted at equilibrium as well as impaired extraction of all analytes at elevated sampling temperatures. Within short HSSPME sampling times and under mild sampling temperatures, Vac-HSSPME yielded linear calibration curves in the range of 1-400 ng g(-1) and, with the exception of fluorene, regression coefficients were found higher than 0.99. The limits of detection for spiked sand samples ranged from 0.003 to 0.233 ng g(-1) and repeatability from 4.3 to 10 %. Finally, the amount of PAHs extracted from spiked soil samples was smaller compared to spiked sand samples, confirming that soil could bind target analytes more strongly and thus decrease the readily available fraction of target analytes.


BLOC-1 complex deficiency alters the targeting of adaptor protein complex-3 cargoes.
Mutational analyses have revealed many genes that are required for proper biogenesis of lysosomes and lysosome-related organelles. The proteins encoded by these genes assemble into five distinct complexes (AP-3, BLOC-1-3, and HOPS) that either sort membrane proteins or interact with SNAREs. Several of these seemingly distinct complexes cause similar phenotypic defects when they are rendered defective by mutation, but the underlying cellular mechanism is not understood. Here, we show that the BLOC-1 complex resides on microvesicles that also contain AP-3 subunits and membrane proteins that are known AP-3 cargoes. Mouse mutants that cause BLOC-1 or AP-3 deficiencies affected the targeting of LAMP1, phosphatidylinositol-4-kinase type II alpha, and VAMP7-TI. VAMP7-TI is an R-SNARE involved in vesicle fusion with late endosomes/lysosomes, and its cellular levels were selectively decreased in cells that were either AP-3- or BLOC-1-deficient. Furthermore, BLOC-1 deficiency selectively altered the subcellular distribution of VAMP7-TI cognate SNAREs. These results indicate that the BLOC-1 and AP-3 protein complexes affect the targeting of SNARE and non-SNARE AP-3 cargoes and suggest a function of the BLOC-1 complex in membrane protein sorting.


Tonsillar hypertrophy and chronic hypoxia.
Three patients with hypertrophied tonsils presented in the Mater Children's Hospital with chronic hypoxia and congestive cardiac failure. The clinical features and physiological basis of this unusual syndrome are discussed. The condition is potentially fatal, but may be completely cured by adenotonsillectomy.


Microwave thermography as a noninvasive assessment of disease activity in inflammatory arthritis.
Microwave thermography is the technique of measuring natural thermal radiation from body tissues of clinical interest. It is safe, requires no control of environmental conditions and exposes the patient to no ionising radiation. In this study, we found that microwave thermography of the knee joints showed weak correlation with some global parameters of disease activity. Bed rest alone for three days had no significant effect on microwave thermographic index (MTI). Intra-articular steroid injection, however, was associated with a marked fall in MTI both in the injected knee and the contra-lateral knee. We conclude that microwave thermography can measure inflammatory activity in the knee joints of patients with inflammatory arthritis and can respond to clinical change brought about by major treatment intervention.


Initial experience of a single referral centre using excimer laser coronary atherectomy-assisted expansion in undilatable stents: Excimer laser in undilatable stents.
Undilatable stents with severe peri-stent calcification are an important cause of target lesion failure and therapeutic options are limited. We report our initial experience with the safety and efficacy of excimer laser coronary atherectomy (ELCA)-assisted expansion of undilatable stents. ELCA was performed with saline, blood and contrast-enhanced trains. All lesions were post-dilated at high pressures and treated with a drug-eluting balloon. Thirty-one lesions with undilatable stents were included at a single centre with experienced operators from March 2016 to February 2021. The mean number of prior procedures for in-stent restenosis was three and 14 lesions had multiple layers of stent. Procedural success (>50% increase in minimal stent diameter [MSD]) and adequate stent expansion (MSD >70% of reference vessel diameter) was achieved in all lesions. At six-month follow-up (N=26 lesions), there were six periprocedural myocardial infarctions due to slow flow, two cardiac deaths and one target lesion revascularisation. There were no perforations. Our niche experience at a centre with experienced operators demonstrated that ELCA led to larger final lumen and stent dimensions in highly selected patients with undilatable stents and recurrent restenosis at the cost of relatively frequent slow flow.


[The influence of select pathophysiologic factors of acute pancreatitis on erythrocytes' deformability examined in vitro].
Deformability of erythrocytes is essential factor in maintenance of blood flow in microcirculation. Deformability of erythrocytes during sepsis, including severe acute pancreatitis, decreases. The aim of the study was to examine in vitro influence of most common etiological factors and determinants of severity of acute pancreatitis on deformability of erythrocytes measured by means of laser diffractometry. We found that ethanol, bilirubin, sodium taurocholate have no effect on deformability of erythrocytes. Trypsin revealed positive effect on deformability of erythrocytes. Acetaldehyde and superoxide anion decrease deformability of erythrocytes in low shear stresses, whereas platelet activating factor and lipopolysaccharide are potent to decrease the deformability of erythrocytes mainly in high shear stresses.


Manganese superoxide dismutase polymorphism, prediagnostic antioxidant status, and risk of clinical significant prostate cancer.
Oxidative stress may enhance prostatic carcinogenesis. A polymorphism [valine (V) --> alanine (A)] of manganese superoxide dismutase (MnSOD), the primary antioxidant enzyme in mitochondria, has been recently associated with prostate cancer. We examined the relationship between prostate cancer and the MnSOD polymorphism and its interactions with baseline plasma antioxidant levels (selenium, lycopene, and alpha-tocopherol) and beta-carotene treatment among 567 cases and 764 controls nested in the prospective Physicians' Health Study. We found little overall association between MnSOD polymorphism and prostate cancer risk; however, this polymorphism significantly modified risk of prostate cancer associated with prediagnostic plasma antioxidants (P(interaction) > or = 0.05). Among men with the AA genotype, high selenium level (4th versus 1st quartile) was associated with a relative risk (RR) of 0.3 [95% confidence interval (CI), 0.2-0.7] for total prostate cancer; for clinically aggressive prostate cancer, the RR was 0.2 (95% CI, 0.1-0.5). In contrast, among men with the VV/VA genotype, the RRs were 0.6 (0.4-1.0) and 0.7 (0.4-1.2) for total and clinically aggressive prostate cancer. These patterns were similar for lycopene and alpha-tocopherol and were particularly strong when these antioxidants and selenium were combined; men with the AA genotype had a 10-fold gradient in risk for aggressive prostate cancer across quartiles of antioxidant status. Men with AA genotype who were randomly assigned to beta-carotene treatment (versus placebo) had a RR of 0.6 (95% CI, 0.2-0.9; P(interaction) = 0.03) for fatal prostate cancer, but no significant association was observed in men with the VV/VA genotype. Both endogenous and exogenous antioxidants play an important and interdependent role in preventing clinically significant prostate cancer.


Vigabatrin as First-Line Treatment for Infantile Spasms Not Related to Tuberous Sclerosis Complex.
Infantile spasms are a rare, catastrophic, age-specific seizure disorder of infancy. Adrenocorticotropic hormone or vigabatrin have been recommended for the short-term treatment of infantile spasms by the Child Neurology Society/American Academy of Neurology practice parameter. This retrospective study reviewed the Hospital for Sick Children's experience with the short-term efficacy of vigabatrin as first-line treatment for infantile spasms not related to tuberous sclerosis complex. We performed a single-center, retrospective analysis of all cases of newly diagnosed infantile spasms between January 2010 and September 2013. Duration of follow-up was at least 6 months from treatment initiation. Eighteen of the 61 infants (30%) had a clinical and electrographic response to vigabatrin therapy within 4 weeks of treatment. Of the vigabatrin responders, 2/18 (11%) relapsed. At final follow-up after initiation of vigabatrin therapy, 17/61 (27%) of the vigabatrin responders were free of all clinical seizure types. Normal development at the time of infantile spasms diagnosis was statistically associated with vigabatrin response. These findings do not support our hypothesis that vigabatrin is effective as first-line, short-term treatment of infantile spasms in non-tuberous sclerosis complex patients. However, when used in this setting, vigabatrin is most effective in children with normal development at the time of diagnosis. These findings may assist clinicians in the optimal treatment choice for children at the first presentation of infantile spasms not related to tuberous sclerosis complex.