Reducing underage alcohol and tobacco use: evidence from the introduction of vertical identification cards.
From 1994 to 2009, forty-three states changed the design of their driver's license/state identification cards in an effort to reduce underage access to and consumption of alcohol and tobacco. In these states, individuals under the age of 21 are issued licenses that are vertically oriented, whereas licenses for individuals 21 and older retain a traditional horizontal shape. This paper examines the effect of this design change on underage alcohol and tobacco use. Using a difference-in-differences methodology, we find a reduction in drinking and smoking for 16 year olds. These results are upheld in a triple difference model that uses a within state control group of teens that did not receive a vertical license to control for state-specific unobserved factors. Interestingly, we find that the effects of the design change are concentrated in the 1-2 years after a state begins issuing vertical licenses. We consider various explanations for our findings: teen learning, the availability of false identification, and changes in retailer behavior.


Higher Benefit for Greater Need: Understanding Changes in Mental Well-being of Young Adults Following the ACA Dependent Coverage Mandate.
Beginning in late 2010, private health insurance plans were required to allow dependents up to age 26 to remain on a parent's plan. Known as the dependent coverage or young adult mandate, this provision increased coverage substantially within the group of 19-25 year-olds affected by the policy change. Subsequent work evaluating whether increased coverage had a positive effect on mental health found mild improvements in self-reported mental health. This work focused exclusively on average effects among young adults in the years after the policy change, leaving open the question of how young adults fared depending on where they reside in terms of the distribution of risk for mental health issues. We assess the effects of the dependent coverage mandate on young adult mental well-being focusing on the distribution of mental health issues. We seek to understand how potential improvements (or degradations) differ across the entire risk profile. Gains among individuals who are at low risk for severe mental health issues may send a far different signal than gains among those with higher risks. Using MEPS data from 2006 through 2013, we use quantile regression within a difference-in-differences design to compare pre/post outcomes across the distribution of risk for young adults ages 23-25 affected by the mandate to 27-29 year-olds not affected by the mandate. Further, we evaluate differences in the effect of the mandate by sex, given well-known disparities in incidence and prevalence of mental illness between men and women. To gauge the effects of the mandate on mental health, we use the Mental Component Score measure within the MEPS, ideal for our quantile regression given the broad range of scores. The key premise in our evaluation is that individuals with higher risks for mental health problems due to biological or socioeconomics factors are more likely to rank at locations of the mental health score distribution indicating worse outcomes. We find significant improvements in self-reported mental health in the 23-25 year-old group following the mandate. However, the gains were not equal across the risk distribution. For individuals at the 0.1 quantile (worse self-reported mental health), the improvement in MCS scores was significant, a 6.1% increase compared to the pre-mandate baseline at that quantile. Effects were smaller but still significant at the median but there was no apparent effect for those that were at higher levels of self-reported mental health. Our results also suggest improvements for women (+9% relative to baseline at the 0.1 quantile, e.g.) but limited evidence of an effect for men. The finding that increased insurance coverage led to improved self-reported mental health foremost for young adults with the highest risk of mental health problems is encouraging. However, the mechanism for this effect is unclear and in need of further study. Whether improvements in the mental health status of the population depend more on increased access to services or derive primarily from improved financial security is an important research area.


Generation of 35 fs, 20 μJ, GHz pulse burst by hybrid fiber amplification technique.
We have proposed and demonstrated the generation of a high-energy, ultrashort pulse duration, GHz pulse burst polarization-maintaining fiber amplification system that utilizes both chirped-pulse amplification and self-similar amplification techniques. Such hybrid fiber amplification system produces 22 μJ-energy bursts of 200 pulses with a 1.02-GHz intra-burst pulse repetition rate and a 1-MHz inter-burst repetition rate. The center wavelength of the amplified compressed pulse is 1065 nm, with a 3 dB spectral bandwidth of 65 nm. The pulse duration of optimal compression is ∼35 fs, which represents the shortest pulse duration reported to date for any multi-microjoule class amplification system with a repetition rate at the GHz level. At the same time, only common double-cladding Yb3+-doped fiber is used as the gain fiber, without any large-mode-area Yb3+-doped photonic crystal fiber, makes the system compact and reliable by the simple fusion operation.


The complete mitochondrial genome of the Chinese peacock, Papilio bianor (Insecta: Lepidoptera: Papilionidae).
The sequence of the mitochondrial genome (mitogenome) of the Chinese peacock Papilio bianor has been completed. It is 15,332 bp with an A + T content of 80.6% and contains the typical complement of 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA), and 2 ribosomal RNA (rRNA) genes. Comparison of this sequence with the sequences of the other completely sequenced insect mitogenomes showed that the gene order and orientation are basically identical to that of Drosophila yakuba with the exception of the rearrangement of one tRNA gene. Nucleotide composition is very similar with other insects, showing a high bias toward A + T.


Dairy Protein Supplementation Modulates the Human Skeletal Muscle microRNA Response to Lower Limb Immobilization.
Limb immobilization results in a rapid loss of muscle size and strength. The resultant alterations in signaling pathways governing myogenesis, catabolism, and mitochondrial biogenesis are likely to include posttranscriptional regulation mediated by altered microRNAs (miRNAs). Given that protein ingestion exerts an anabolic action and may act as a countermeasure to mitigate muscle loss with immobilization, it is important to examine miRNA in this context. The objective of the study is therefore to characterize the vastus lateralis miRNA response to 14 days of disuse in males (45-60 years) randomized to receive supplementation with 20 g d-1 of dairy protein (n = 12) or isocaloric carbohydrate placebo (n = 13). Biopsies are collected before and after a 2-week immobilization period. Of the 24 miRNAs previously identified in myogenic regulation, seven (miR-133a, -206, -15a, -451a, -126, -208b, and let-7e) are increased with immobilization irrespective of group; five (miR-16, -494, let-7a, -7c, and 7d) increased only in the carbohydrate group; and eight (miR-1, -486, -23a, -23b, -26a, -148b, let-7b, and -7g) are divergently expressed between groups (suppressed with protein). The ability of protein supplementation to differentially regulate miRNAs involved in key muscle regulatory pathways following short-term limb immobilization reflects potential protective function in mitigating muscle loss during limb immobilization.


Pulmonary artery banding for hemi-truncus arteriosus in adulthood.
Anomalous origin of the right pulmonary artery from the ascending aorta, also called hemi-truncus arteriosus, is a rare congenital cardiovascular malformation. Around 95% of the reported cases with hemi-truncus arteriosus present in infancy. We describe a case of a 41-year-old female presenting with a long-standing history of intermittent haemoptysis. Cardiac catheterisation confirmed the diagnosis of hemi-truncus arteriosus. The definitive treatment of hemi-truncus arteriosus in adulthood is an extremely high-risk procedure. We decided to perform palliative right pulmonary artery banding which represents a simple but effective procedure for the management of this complex condition.


Vertical foliage distribution determines the radial pattern of sap flux density in Picea abies.
Understanding the causes determining the radial pattern of sap flux density is important both for improving knowledge of sapwood functioning and for up-scaling sap flow measurements to canopy transpiration and ecosystem water use. To investigate the anatomical connection between whorls and annual sapwood rings, pruning-induced variation in the radial pattern of sap flux density was monitored with Granier probes in a 35-year-old Picea abies (L.) Karst tree that was pruned from the crown bottom up. Modifications in the radial pattern of sap flux density were quantified by a shape index (SI), which varies with the relative contribution of the outer and inner sapwood to tree transpiration. The SI progressively diminished during bottom up pruning, indicating a significant reduction in sap flow contribution of the inner sapwood. Results suggest that the radial pattern of sap flux density depends mainly on the vertical distribution of foliage in the crown, with lower shaded branches hydraulically connected with inner sapwood and upper branches connected with the outer rings.


Management and treatment of glycogenosis type II.
Glycogenosis type II is a multisystem disorder that requires management by a multidisciplinary team. The team should include several specialists, such as a metabolic disease specialist or biochemical geneticist, cardiologist, pulmonologist, neurologist, neuromuscular specialist, intensivist, orthopedist, respiratory therapist, physical therapist, occupational therapist, otolaryngologist speech therapist, audiologist, genetic counselor, and a metabolic dietician, who, as a team, will be capable of addressing the different manifestations of the condition. Aspects of functional assessment, rehabilitation, nutritional management, care coordination, nursing, genetic counseling, prenatal diagnosis, and screening are discussed in this article. In addition, treatment of glycogenosis type II is reviewed with attention to emerging therapeutic options.


CT quantification of pneumonia lesions in early days predicts progression to severe illness in a cohort of COVID-19 patients.
Rationale: Some patients with coronavirus disease 2019 (COVID-19) rapidly develop respiratory failure or even die, underscoring the need for early identification of patients at elevated risk of severe illness. This study aims to quantify pneumonia lesions by computed tomography (CT) in the early days to predict progression to severe illness in a cohort of COVID-19 patients. Methods: This retrospective cohort study included confirmed COVID-19 patients. Three quantitative CT features of pneumonia lesions were automatically calculated using artificial intelligence algorithms, representing the percentages of ground-glass opacity volume (PGV), semi-consolidation volume (PSV), and consolidation volume (PCV) in both lungs. CT features, acute physiology and chronic health evaluation II (APACHE-II) score, neutrophil-to-lymphocyte ratio (NLR), and d-dimer, on day 0 (hospital admission) and day 4, were collected to predict the occurrence of severe illness within a 28-day follow-up using both logistic regression and Cox proportional hazard models. Results: We included 134 patients, of whom 19 (14.2%) developed any severe illness. CT features on day 0 and day 4, as well as their changes from day 0 to day 4, showed predictive capability. Changes in CT features from day 0 to day 4 performed the best in the prediction (area under the receiver operating characteristic curve = 0.93, 95% confidence interval [CI] 0.87~0.99; C-index=0.88, 95% CI 0.81~0.95). The hazard ratios of PGV and PCV were 1.39 (95% CI 1.05~1.84, P=0.023) and 1.67 (95% CI 1.17~2.38, P=0.005), respectively. CT features, adjusted for age and gender, on day 4 and in terms of changes from day 0 to day 4 outperformed APACHE-II, NLR, and d-dimer. Conclusions: CT quantification of pneumonia lesions can early and non-invasively predict the progression to severe illness, providing a promising prognostic indicator for clinical management of COVID-19.


Mental health, coping and family-functioning in parents of young people with obsessive-compulsive disorder and with anxiety disorders.
To compare mental health, coping and family-functioning in parents of young people with obsessive-compulsive disorder (OCD), anxiety disorders, and no known mental health problems. Parents of young people with OCD (N = 28), other anxiety disorders (N = 28), and no known mental health problems (N = 62) completed the Brief Symptom Inventory (Derogatis, 1993), the Coping Responses Inventory (Moos, 1990), and the McMaster family assessment device (Epstein, Baldwin, & Bishop, 1983). Parents of children with OCD and anxiety disorders had poorer mental health and used more avoidant coping than parents of non-clinical children. There were no group differences in family-functioning. The similarities across the parents of clinically referred children suggest that there is a case for encouraging active parental involvement in the treatment of OCD in young people.


p120-catenin phosphorylation status alters E-cadherin mediated cell adhesion and ability of tumor cells to metastasize.
p120-catenin is considered to be a tumor suppressor because it stabilizes E-cadherin levels at the cell surface. p120-catenin phosphorylation is increased in several types of cancer, but the role of phosphorylation in cancer is unknown. The phosphorylation state of p120-catenin is important in controlling E-cadherin homophilic binding strength which maintains epithelial junctions. Because decreased cell-cell adhesion is associated with increased cancer metastasis we hypothesize that p120-catenin phosphorylation at specific Serine and Threonine residues alters the E-cadherin binding strength between tumor cells and thereby affect the ability of tumor cells to leave the primary tumor and metastasize to distant sites. In this study we show that expression of the p120-catenin phosphorylation dead mutant, by converting six Serine and Threonine sites to Alanine, leads to enhanced E-cadherin adhesive binding strength in tumor cells. We observed a decrease in the ability of tumor cells expressing the p120-catenin phosphorylation mutant to migrate and invade using in-vitro models of cancer progression. Further, tumor cells expressing the phosphorylation mutant form of p120-catenin demonstrated a decrease in ability to metastasize to the lungs using an in-vivo orthotopic mammary fat pad injection model of breast cancer development and metastasis. This suggests that regulation of p120-catenin phosphorylation at the cell surface is important in mediating cell-adhesion, thereby impacting cancer progression and metastasis.


HIV type 1 variants transmitted to women in Kenya require the CCR5 coreceptor for entry, regardless of the genetic complexity of the infecting virus.
Although nearly half of the HIV-1-infected adults in the world are women, little is known about the virologic determinants of transmission to women. Studies suggest that women are frequently infected with multiple HIV-1 genotypes, whereas men are infected with a single genotype. In the current study, we assessed whether the diverse HIV-1 genomes present at the time of infection in women encode viruses that have diverse coreceptor specificities. For this purpose, we defined the coreceptor requirements of viruses found in recently infected Kenyan women, three of whom had multiple viral genotypes and the remaining two of whom had a single genotype. Full-length envelope clones were amplified directly from blood and the dominant genotypes were identified. Envelope clones derived from all five women were able to pseudotype infectious particles competent to infect cells expressing CCR5, but not cells expressing only CXCR4. Thus, regardless of viral complexity at the time of infection, the viruses present at early stages of HIV-1 infection in women use CCR5, suggesting that cells expressing CCR5 are important targets for heterosexual HIV-1 transmission to women.


[Pharmacokinetics of rectal absorption of amodiaquine in rabbits].
Experimental investigations were made in rabbits to examine the rectal absorption of antimalarial amodiaquine [AQ]. To calculate the pharmacokinetic parameters AQ was administered orally (20 mg/kg) and rectally (20 mg/kg) to the animals and the blood concentrations of the drug were controlled during a time interval of 120 hours with help of spectrofluorometric method. The pharmacokinetic analysis was performed by use of two-compartment open model system. Time-concentration curves were fitted to the data in experiments by computer program (MEDUSA) and the kinetic parameters were given in the Tables 1. and 2. It was established that rectally administered AQ resorbed not only from solution but from suppository, too. The bioavailability of the rectal preparations relating to the oral dosage form was 63.35% and 68.70%, respectively. The most appropriate drug delivery system for rectal application had been selected previously.


The complete nucleotide sequence of a new bipartite begomovirus from Brazil infecting Abutilon.
The complete nucleotide sequence of Abutilon mosaic Brazil virus (AbMBV), a new bipartite begomovirus from Bahia, Brazil, is described and analyzed phylogenetically. Its DNA A is most closely related to those of Sida-infecting begomoviruses from Brazil and forms a phylogenetic cluster with pepper- and Euphorbia-infecting begomoviruses from Central America. The DNA B component forms a cluster with different Sida- and okra-infecting begomoviruses from Brazil. Both components are distinct from those of the classical Abutilon mosaic virus originating from the West Indies. AbMBV is transmissible to Nicotiana benthamiana and Malva parviflora by biolistics of rolling-circle amplification products and induces characteristic mosaic and vein-clearing symptoms in M. parviflora.


Present but not accounted for: exploring the sexual risk practices and intervention needs of nonheterosexually identified women in a prevention program for women with HIV/AIDS.
Nonheterosexually identified (NHI) women may be present, but not accounted for, in HIV and sexually transmitted infection (STI) prevention interventions. This study used quantitative and qualitative methods to examine the sexual risk behaviors and intervention needs of NHI women in Protect and Respect, a safer sex intervention for HIV-positive women. Study participants (n=32) were predominantly Black, low income, and between 28 and 51 years old. Although NHI participants were more likely than heterosexual participants (p < .05) to report obtaining their income from sex work, hustling, or selling drugs; and having a higher median number of male sex partners, qualitative analyses revealed that the intervention often neglected NHI women's experiences and unique safer sex needs. Heterosexist HIV and STI prevention programs may hinder NHI women's ability to protect themselves and their partners from reinfection and infection respectively. We discuss the implications of our research for future HIV/AIDS and STI research, services and interventions for NHI women.


Highly enantioselective, intermolecular hydroamination of allenyl esters catalyzed by bifunctional phosphinothioureas.
Bifunctional phosphinothiourea catalysts have been developed successfully for the highly regio- and enantioselective γ-hydroamination of allenyl and propargyl esters with N-methoxy carbamate nucleophiles to yield α,β-unsaturated γ-amino acid ester products. In the case of propargyl ester substrates, the reaction proceeds through reversible phosphinothiourea-catalyzed isomerization to the corresponding allenyl ester. The high enantioselectivity of the process is attributed to a cooperative conjugate addition of a thiourea-bound carbamate anion to a vinyl phosphonium ion resulting from covalent activation of the allenyl ester substrate.


Evaluation of milk sample fractions for characterization of milk microbiota from healthy and clinical mastitis cows.
Amplicon sequencing technique has been increasingly applied to the clinical setting as a sensitive diagnostic tool. Therefore, it is of great importance to develop a DNA extraction method that accurate isolates DNA from complex host-associated microbiota. Given the multifactorial etiology of clinical mastitis and the diversified lifestyle of bacterial species harboring in milk, here four distinct milk sample fractions: raw whole milk, milk fat, casein-pellet, and casein-pellet + fat from healthy cows and cows with clinical mastitis, were subjected to bead-beating DNA extraction, followed by high-throughput sequencing. We aimed to identify the best approach for characterization of the milk microbiota and detection of mastitis pathogens (Klebsiella spp., Streptococcus spp. and Escherichia coli). DNA from each milk fraction tested was extracted by two commercial kits, which include physical, mechanical and chemical lysis; in total 280 DNA samples from 35 cows were analyzed. Milk-health-status were categorized into four groups (healthy group; E. coli-mastitis group; Klebsiella spp.-mastitis group; and Streptococcus spp.-mastitis group). Bacterial phyla and families were described for each milk-health-status group across milk sample fractions and DNA extraction kits. For the mastitis groups the relative abundance of f__Enterobacteriaceae and f__Streptococcaceae were compared to determine the efficacy of procedures in detecting the mastitis pathogens. The four milk fractions used allowed efficiently and uniformly detection of the causative agent of mastitis. Only 27% of the families detected in healthy milk were shared among the samples extracted from all fractions of milk samples; followed by 3, 4, and 12% for the samples from E. coli-mastitis, Klebsiella spp.-mastitis and Streptococcus spp-mastitis, respectively. However, the shared families comprised a mean relative abundance greater than 85%, regardless of milk-health-status, milk fraction and DNA isolation method. Taxonomic data at the family level showed that sequences from mastitis milk samples cultured positive for E. coli and Klebsiella spp. were predominantly affiliated with f__Enterobacteriaceae, while for Streptococcus spp. were dominated by f__Streptococcacea, followed by f__Pseudomonadaceae and f__Enterococcaceae. Microbial community analysis revealed that most of the microbial community composition corresponded to milk bacterial species irrespective of the DNA isolation method and milk fraction evaluated.


Synthetic speech comprehension: a comparison of children with normal and impaired language skills.
This study compared the ability of children with normal language (NL) and children with specific language impairment (SLI) to comprehend natural speech and DECtalk synthetic speech by using a sentence verification task. The effect of listening practice on subjects' ability to comprehend both types of speech also was investigated. Subjects were matched for age and sex. Mean nonverbal intelligence scores of the groups did not differ significantly. Results showed that DECtalk was significantly more difficult for all subjects to comprehend than was natural speech and false sentences were significantly more difficult to comprehend than were true sentences. Response latencies shortened significantly from time 1 to time 2 for all subjects. Subjects with SLI had significantly more difficulty comprehending both natural and synthetic speech than did subjects with NL. Implications these results might have for theories of the underlying cause of specific language impairment are discussed.


Immunologic and metabolic biomarkers of β-cell destruction in the diagnosis of type 1 diabetes.
Type 1 diabetes (T1D), also known as insulin-dependent diabetes mellitus, is a chronic disorder that results from autoimmune destruction of insulin-producing β cells in the islets of Langerhans within the pancreas ( Atkinson and Maclaren 1994). This disease becomes clinically apparent only after significant destruction of the β-cell mass, which reduces the ability to maintain glycemic control and metabolic function. In addition, it continues for years after clinical onset until, generally, there is complete destruction of insulin secretory capacity. Because prevention and therapy strategies are targeted to this pathologic process, it becomes imperative to have methods with which it can be monitored. This work discusses current research-based approaches to monitor the autoimmunity and metabolic function in T1D patients and their potential for widespread clinical application.


Acceptability of bone antiresorptive therapy among HIV-infected adults at different stages of antiretroviral therapy.
Both HIV infection and antiretroviral therapy (ART) are associated with significant decreases in bone mineral density (BMD) and increased fracture rates. To prepare for a randomized controlled trial of prophylactic bone antiresorptive therapy during ART initiation, we assessed the acceptability of this strategy, bone health knowledge, and fracture risk among HIV-infected adults. HIV-infected adults with no history of osteoporosis were recruited from one tertiary and one primary care HIV clinic. Participants completed a questionnaire and underwent chart review. The primary outcome was the proportion of respondents expressing interest in taking prophylactic bone antiresorptive therapy in conjunction with ART. Of 112 respondents, 25.0% were ART naïve, 23.2% had been taking ART for ≤1 year, and 51.8% had been taking ART for >1 year. Half (51.9%) indicated interest in taking short-course prophylactic bone antiresorptive therapy; this did not differ by ART status (53.6% among ART-naïve, 51.3% among ART-treated; P=0.84, chi-square test). In exploratory multivariable analysis adjusted for ART status, a greater number of pills taken per day was positively associated with this outcome (adjusted odds ratio [OR] =1.12 per pill, 95% confidence limit [CL] =1.01, 1.25), while male sex was inversely associated (adjusted OR =0.05, 95% CL =0.01, 0.24). Among those willing to take therapy, most (80.4%) were willing to do so for "as long as needed" and preferred weekly dosing (70.9%) to daily dosing (12.7%). Half of this sample would be willing to take bone antiresorptive therapy together with ART, with preferences for weekly dosing and for whatever duration may be required. These data will inform the design of future trials to protect bone health in HIV.


Synchrotron radiation microdiffraction of ballistic molten wax microdrops.
Using stroboscopic techniques, diffraction patterns of ballistic paraffin wax microdrops have been observed. The microdrops, generated by a high-temperature ink-jet system, travel through the 1 mum synchrotron radiation beam with a speed of about 1.4 m/s. Diffraction patterns were recorded in flight by a charge couple device with a microchannel plate image intensifier stage, which was activated with the microdrop generation frequency of 1000 Hz during 2 mus. The data show liquid microdrops with a constant temperature up to 8 mm from the ink-jet system capillary exit. The general technique could be adapted for studying fast structural processes, such as protein conformational changes in aqueous microdrops.


Pressure-Modulated Dissolution Behavior of LLM-105 Crystals in High-Temperature Water.
The exploration of the microstructural evolution and reaction kinetics of energetic materials with high-temperature and high-pressure water contributes to the understanding of their microscopic physicochemical origin, which can provide critical experimental data for the use of energetic materials. As a promising high-energy and insensitive energetic material, LLM-105 has been investigated under extreme conditions such as high pressure and high temperature. However, little information is available about the effect of water on LLM-105 under high pressure and high temperature. In this work, the interaction between LLM-105 and water under HP-HT was investigated in detail. As a result, the dissolving behavior of LLM-105 in water under high pressure and high temperature is related to the initial pressure. When the initial pressure is less than 1 GPa, LLM-105 crystals are dissolved in high-temperature water; when the initial pressure is above 1 GPa, LLM-105 particles are only decomposed in high-temperature water. When the solution is saturated at a high temperature, recrystallization of the LLM-105 sample appears in the solution. High pressure hindered the dissolution process of the sample in HP-HT water because the interaction between the solute and the solvent was weakened by high pressure. The initial pressure is one of the significant parameters that determines whether LLM-105 crystals can be dissolved in high-temperature water. More importantly, water under high pressure and high temperature can not only act as a solvent when dissolving the samples but also act as a catalyst to accelerate the decomposition process. In addition, the HP-HT water reduced the decomposition temperature of the LLM-105 crystal to a large extent. The research in this paper not only provides insights into the interaction between LLM-105 and water but also contributes to the performance of energetic materials under extreme conditions and their practical applications in complex conditions.


Pertuzumab and trastuzumab: the rationale way to synergy.
It has now been 15 years since the HER2-targeted monoclonal antibody trastuzumab was introduced in clinical and revolutionized the treatment of HER2-positive breast cancer patients. Despite this achievement, most patients with HER2-positive metastatic breast cancer still show progression of their disease, highlighting the need for new therapies. The continuous interest in novel targeted agents led to the development of pertuzumab, the first in a new class of agents, the HER dimerization inhibitors. Pertuzumab is a novel recombinant humanized antibody directed against extracellular domain II of HER2 protein that is required for the heterodimerization of HER2 with other HER receptors, leading to the activation of downstream signalling pathways. Pertuzumab combined with trastuzumab plus docetaxel was approved for the first-line treatment of patients with HER2-positive metastatic breast cancer and is currently used as a standard of care in this indication. In the neoadjuvant setting, the drug was granted FDA-accelerated approval in 2013. Pertuzumab is also being evaluated in the adjuvant setting. The potential of pertuzumab relies in the dual complete blockade of the HER2/3 axis when administered with trastuzumab. This paper synthetises preclinical and clinical data on pertuzumab and highlights the mechanisms underlying the synergistic activity of the combination pertuzumab-trastuzumab which are essentially due to their complementary mode of action.


Absorption, distribution, and excretion of 5-azacytidine (NSC-102816) in man.
Patients with advanced cancer were given 5-azacytidine labeled at position 4 with radioactive carbon (14C) by either the intravenous (iv) or subcutaneous (sc) route. Absorption of the drug from the sc injection site was rapid and peak plasma levels were attained within one-half hour. Within 2 hours, the plasma level of radioactivity was approximately equal to that noted in the patients treated iv. The plasma half-life after iv injection was 3.5 hours; after sc administration, the plasma half-life was 4.2 hours. Patients receiving the drug sc excreted less drug in the urine than did those receiving the drug iv. No radioactivity was detected in the expired carbon dioxide when the drug was given by either route. Drug uptake into tumor tissue was always greater than uptake into surrounding normal tissue. The highest concentrations of radioactivity in the tissues were achieved when the drug had been given iv. Traces of radioactivity were still detectable in the tissues for as long as 6 days after administration of the drug. The incorporation of radioactivity into tumor RNA but not into DNA was demonstrated. The maximum level of radioactivity detected in the spinal fluid was equivalent to 0.2 microg of 5-azacytidine per milliliter of fluid.


Theoretical analysis of residual dipolar coupling patterns in regular secondary structures of proteins.
A new approach to the interpretation of residual dipolar couplings for the regular secondary structures of proteins is presented. This paper deals with the analysis of the steric and chiral requirements of protein secondary structures and establishes a quantitative correlation between structure periodicity and the experimental values of the backbone residual dipolar couplings. Building on the recent interpretation of the periodicity of residual dipolar couplings in alpha-helices (i.e., "dipolar waves"), a general parametric equation for fitting the residual dipolar couplings of any regular secondary structure is derived. This equation interprets the modulation of the residual dipolar couplings' periodicity in terms of the secondary structure orientation with respect to an arbitrary reference frame, laying the groundwork for using backbone residual dipolar couplings as a fast tool for determining protein folding by NMR spectroscopy.


Visualization of in Vivo Hydrogen Sulfide Production by a Bioluminescence Probe in Cancer Cells and Nude Mice.
Hydrogen sulfide (H2S) has emerged as an exciting endogenous gasotransmitter in addition to nitric oxide and carbon monoxide. However, its precise measurement in living cells and animals remains a challenge. In this study, a novel bioluminescence H2S probe was designed and synthesized by modifying the 6'-amino group of d-aminoluciferin into a 6'-azido group, which was highly selective against other reactive sulfur, nitrogen, and oxygen species. Our H2S probe azidoluciferin sensitively reacted with H2S to release d-aminoluciferin with a strong bioluminescence signal. On the basis of its high selectivity and sensitivity, the H2S probe was used to detect H2S production in live cancer cells and nude mice. The bioluminescence signal decreased in mice treated with propargylglycine, an inhibitor of H2S, suggesting that our H2S probe can detect endogenous H2S in real time, in vivo. Overall, the excellent sensing properties of the probe combined with its bioimaging capability make it a useful tool to study H2S biological roles.


Long-term monitoring of the Iberian ibex population in the Sierra Nevada of the southeast Iberian Peninsula.
This dataset provides long-term information on the presence of the Iberian ibex (Capra pyrenaica hispanica Schimper, 1848) in Sierra Nevada (SE Iberian Peninsula). Data on the abundance and demographic structure of the Iberian ibex population were compiled over the last three decades. Transects were laid out to record different variables such as the number of individuals sighted, the perpendicular distance of each group of Iberian ibex to the transect line and sex as well as age of individuals in the case of males. These data enabled the calculation of population parameters such as density, sex ratio, birth rate, and age structure. These parameters are key for Iberian ibex conservation and management, given that Sierra Nevada harbours the largest population of this species in the Iberian Peninsula. The data set we present is structured using the Darwin Core biological standard, which contains 3,091 events (582 transect walk events and 2,509 group sighting events), 5,396 occurrences, and 2,502 measurements. The occurrences include the sightings of 11,436 individuals (grouped by sex and age) from 1993 to 2018 in a total of 88 transects distributed along Sierra Nevada, of which 33 have been continuously sampled since 2008.


[Ambulatory anesthesia preoperative evaluation and its effects on anesthetic care].
Adequate preoperative evaluation with sufficient informed consent from patients will lead to safe perioperative anesthetic managements. We started outpatient preoperative evaluation from October, 2002. We investigated 1137 elective cases operated during one year after starting outpatient examination. They were compared with 1083 cases under the previous system. On requests from surgeons we examined the patients and obtained informed consents. Additional examinations and medications were ordered if necessary for adequate preoperative management. Eight hundred and twenty-three cases (72.4%) of 1137 elective operative cases visited our outpatient booth and 86.8% of them visited before admission for the surgery. Patients classified into ASA-PS II-III, 550 cases (66.9%), and their major complications were cardiovascular and pulmonary diseases. Two hundred and ninety seven cases (36.1%) required additional examinations following adequate preoperative management by another consultants. After the introduction of this system, the number of postponing or cancellation of operation after hospitalization was only 1.3% of elective operations. Furthermore the number of patients admitted on the day and the day before the operation significantly increased compared with those under the previous system (27.9 - 45.2%). Our preoperative consulting system will decrease average length of hospital stay and improve the quality control of patients before surgery.


Cost-effective minimally invasive gynecologic surgery: emphasizing surgical efficiency.
The United States has the highest healthcare costs among developed countries. This review evaluates surgical practices and equipment choices during endoscopic hysterectomy, highlighting opportunities for the gynecologic surgeon to reduce costs and maximize surgical efficiency. There are opportunities to economize at every step of the endoscopic hysterectomy. When surgeons are provided education about instrumentation costs, the cost of hysterectomy has been shown to decrease. Colpotomy has been found to be the rate-limiting step in laparoscopic hysterectomy; use of a uterine manipulator likely saves time and money. When evaluating the economic impact of route of surgery, the cost differential between laparoscopic and robotic-assisted hysterectomy has decreased. Robotic-assisted hysterectomy may be more cost-effective in some cases, such as for larger uteri. From a systems-level perspective, dedicating a specific operating room team to the gynecology service can decrease operative time. The gynecologic surgeon is best equipped to control surgery-related costs by making choices that improve surgical efficiency and decrease operating room time. If a costlier piece of equipment leads to a more efficient case, the choice may be more cost-effective. There are multiple systems-level changes that can be implemented to decrease surgery-related costs.


Who experiences endoscopic retrograde cholangiopancreatography after laparoscopic cholecystectomy for symptomatic gallstone disease?
Laparoscopic cholecystectomy (LC) has become a standard treatment of symptomatic gallstone disease. But, some patients suffer from retained common bile duct stones after LC. The aim of this study is to analyze the predicting factors associated with subsequent postoperative endoscopic retrograde cholangiopancreatography (ERCP) after LC. We retrospectively reviewed a database of every LC performed between July 2006 and September 2012. We classify 28 patients who underwent ERCP within 6 months after LC for symptomatic gallstone disease as the ERCP group and 56 patients who underwent LC for symptomatic gallstone disease during same period paired by sex, age, underlying disease, operation history, and body mass index as the control group. To identify risk factor performing postoperative ERCP after LC, we compared admission route, preoperative biochemical liver function test, number of gall stones, gallstone size, adhesion around GB, wall thickening of GB, and existence of acute cholecystitis between the 2 groups. Admission route, preoperative AST, ALT, and ALP, stone size, longer operation time, and acute cholecystitis were identified as risk factors of postoperative ERCP in univariate analyses. But, longer operation time (P = 0.004) and acute cholecystitis (P = 0.048) were identified as independent risk factors of postoperative ERCP in multivariate analyses. The patient who underwent ERCP after LC for symptomatic gallstone disease are more likely experienced longer operation time and acute cholecystitis than the patient who did not undergo ERCP after LC.


Differential considerations of skin tumours with florid vascularisation: report of a solitary giant vascular eccrine spiradenoma.
The authors report the case of an 81-year-old male who presented with a 3-year-history of a bluish, nodular tumour located on the extensor side of his right forearm. Subjective symptoms included tenderness upon palpation and spontaneous haemorrhage. In order to exclude malignant neoplasms, for example, nodular melanoma, metastatic melanoma or angiosarcoma, the tumour was surgically removed and tissue submitted for microscopic examination. Histologically, the authors diagnosed this as giant vascular eccrine spiradenoma, a rare variant of eccrine spiradenoma, which can easily be mistaken for angiomatous lesions due to the haemorrhagic features and florid vascularisation. It is our aim to help clarify the diagnosis and differentiate giant vascular eccrine spiradenoma from other painful cutaneous tumours exhibiting a high degree of vascularisation, for example, angiosarcoma or venous thrombosis, as this case represents one of only seven found in published literature.


NMDA-mediated modulation of dopamine release is modified in rat prefrontal cortex and nucleus accumbens after chronic nicotine treatment.
In this study, we investigate the effects of chronic administration of (-)nicotine on the function of the NMDA-mediated modulation of [(3)H]dopamine (DA) release in rat prefrontal cortex (PFC) and nucleus accumbens (NAc). In the PFC synaptosomes NMDA in a concentration-dependent manner evoked [(3)H]DA release in rats chronically treated with vehicle (14 days) with an EC(50) of 13.1 +/- 2.0 microM. The NMDA-evoked overflow of the [(3)H]DA in PFC nerve endings of rats treated with (-)nicotine was significantly lower (-43%) than in vehicle treated rats. The EC(50) was 9.0 +/- 1.4 microM. Exposure of NAc synaptosomes of rats treated with vehicle to NMDA produced an increase in [(3)H]DA overflow with an EC(50) of 14.5 +/- 5.5 microM. This effect was significantly enhanced in chronically treated animals. The EC(50) was 10.5 +/- 0.5 microM. The K(+)-evoked release of [(3)H]DA was not modified by the (-)nicotine administration. Both the changes of the NMDA-evoked [(3)H]DA overflow in the NAc and PFC disappeared after 14 days withdrawal. The results show that chronic (-)nicotine differentially affects the NMDA-mediated [(3)H]DA release in the PFC and NAc of the rat.


Variability in post-operative fluid and electrolyte prescription.
A four-week retrospective survey of intravenous fluid and electrolyte prescriptions on post-operative surgical patients revealed wide variability in fluid and electrolyte prescription by medical staff. Median volume of intravenous fluid prescribed was 3000 ml (range 1667-5000). Total sodium prescribed varied from 0 to 770, median 242 mmol/day), with potassium infrequently added (range 0-81, median 0 mmol/day). Patients undergoing emergency surgery were prescribed more sodium than those undergoing routine procedures (p = 0.0403); 0.9% saline was the most common fluid prescribed overall. There was poor correlation between serum electrolyte values and the amounts of electrolytes prescribed. Intravenous fluid prescription should take into account the post-operative stress response which reduces sodium requirements (unless there are other concomitant losses) and increases urinary potassium losses. A suitable post-operative 'maintenance' fluid is 4% dextrose/0.18% saline with 1-2 g potassium chloride, particularly if serum electrolyte levels are not known. Other fluid losses should be replaced with equivalent fluids.


Methotrexate as a single agent and in combination chemotherapy for the treatment of non-Hodgkin's lymphoma of unfavorable histology.
High-dose methotrexate (HDMTX) with citrovorum factor (CF) was used to treat non-Hodgkin's lymphomas either as a single agent in those patients who had previously failed to respond to conventional chemotherapy or in previously untreated patients in combination with bleomycin, Adriamycin, cytoxan, and vincristine. Twenty-five patients who had been previously treated and were refractory to cytotoxic chemotherapy were treated with HDMTX ranging between 1 and 7.5 g/m2 as an iv infusion over 20 to 40 minutes. CF rescue was begun at 24 hours and was maintained orally every 6 hours at 10 mg/m2 for a total of 72 hours. In instances where the 24-hour serum level of MTX was in excess of 1 X 10(-7) M, the dose of CF was increased to 100 mg/m2 every 3 hours until the serum MTX level fell into the appropriate range. Of the 25 patients, 13 (52%) responded. Response duration was brief, lasting a median of 2 months. Three patients had prolonged control lasting 13, 16, and 21 months respectively. Five of the six patients with central nervous system (CNS) involvement have demonstrated marked clearing of spinal fluid abnormalities and reversion of brain scans toward normal. Fifty-four previously untreated patients with diffuse histiocytic or undifferentiated lymphoma have been treated with combination chemotherapy including MTX given at the level of 3 g/m2 on day 14 of a cycle. Bleomycin, Adriamycin, cyclophosphamide, and vincristine are given on Day 1 of the cycle. Dexamethasone 6 mg/m2 is given orally on Days 1 through 5. The cycle is repeated on Day 21, and patients are treated with 10 such cycles for a total of 30 weeks. Of our 42 patients who are evaluable for response and who have completed chemotherapy, 33 (78%) achieved a complete remission (CR). A total of seven patients have relapsed. Only one patient in the CR group has relapsed in the CNS after the cessation of chemotherapy.


B Cell-Based Cancer Immunotherapy.
B cells are not only producers of antibodies, but also contribute to immune regulation or act as potent antigen-presenting cells. The potential of B cells for cellular therapy is still largely underestimated, despite their multiple diverse effector functions. The CD40L/CD40 signaling pathway is the most potent activator of antigen presentation capacity in B lymphocytes. CD40-activated B cells are potent antigen-presenting cells that induce specific T-cell responses in vitro and in vivo. In preclinical cancer models in mice and dogs, CD40-activated B cell-based cancer immunotherapy was able to induce effective antitumor immunity. So far, there have been only few early-stage clinical studies involving B cell-based cancer vaccines. These trials indicate that B cell-based immunotherapy is generally safe and associated with little toxicity. Furthermore, these studies suggest that B-cell immunotherapy can elicit antitumor T-cell responses. Alongside the recent advances in cellular therapies in general, major obstacles for generation of good manufacturing practice-manufactured B-cell immunotherapies have been overcome. Thus, a first clinical trial involving CD40-activated B cells might be in reach.


Platelet activation of mechanical versus bioprosthetic heart valves during systole.
Thrombus formation is a major concern for recipients of mechanical heart valves (MHVs), which requires them to take anticoagulant drugs for the rest of their lives. Bioprosthetic heart valves (BHVs) do not require life-long anticoagulant therapy but deteriorate after 10-15years. The thrombus formation is initiated by the platelet activation which is thought to be mainly generated in MHVs by the flow through the hinge and the leakage flow during the diastole. However, our results show that the activation in the bulk flow during the systole phase might play an essential role as well. This is based on our results obtained by comparing the thrombogenic performance of a MHV and a BHV (as control) in terms of shear induced platelet activation under exactly the same conditions. Three different mathematical activation models including linear level of activation, damage accumulation, and Soares model are tested to quantify the platelet activation during systole using the previous simulations of the flow through MHV and BHV in a straight aorta under the same physiologic flow conditions. Results indicate that the platelet activation in the MHV at the beginning of the systole phase is slightly less than the BHV. However, at the end of the systole phase the platelet activation by the bulk flow for the MHV is several folds (1.41, 5.12, and 2.81 for linear level of activation, damage accumulation, and Soares model, respectively) higher than the BHV for all tested platelet activation models.


Changes of intracellular Ca2+ in quercetin-induced autophagy progression.
Quercetin was previously reported to exhibit significant anti-proliferative activities, and its major effect on tumors was to induce cell apoptosis or autophagy. However, the specific mechanism remains controversial. In this study, autophagy induced by quercetin was determined with various methods. Intracellular Ca2+ ([Ca2+]i) was measured after being incubated with Fluo-3 acetoxymethyl (AM). At the same time, the relationship between the intracellular Ca2+ and autophagy induced by quercetin was further analyzed. These results showed that autophagy induced by quercetin (0-50 µg/ml) in HepG2 cells was in a dose-dependent manner. Meanwhile, when autophagy was induced by quercetin, [Ca2+]i was significantly increased. And after being incubated with calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N-tetraacetic acid-AM, autophagy was suppressed, which implied that [Ca2+]i elevation appeared to be the cause for autophagy induction. These results suggested that calcium from intracellular calcium storage may play an important role in quercetin-induced autophagy.


Molecular Characteristics of Choledochal Cysts in Children: Transcriptome Sequencing.
A choledochal cyst (CC) is a common congenital biliary disease in children, yet the underlying molecular bases for the cystic and fusiform clinical subtypes are unknown. RNA sequencing (RNA-seq) has been performed on 22 high-quality CC samples, including 12 cystic CC and 10 fusiform CC samples, to search for molecular features. Weighted gene co-expression network analysis (WGCNA) was performed to identify key modules associated with clinical subtypes. Bioinformatic analyses were conducted to elucidate potential mechanisms. Then, we constructed protein-protein interaction (PPI) networks to identify candidate hub genes related to CC. Finally, we used the support vector machine (SVM) to eliminate redundant features and screen out the hub genes. The selected gene expression was determined in CC patients through quantitative real-time polymerase chain reaction (PCR). A total of 6,463 genes were found to be aberrantly expressed between cystic CC and fusiform CC. Twelve co-expression modules that correlated with clinical subtypes of CC were identified and assigned representative colors. Among the 12 modules, the blue module was considered the key module. Two functionally distinct sets of dysregulated genes have been identified in two major subtypes, metabolism-related genes in cystic CC and immune-related genes in fusiform CC. A total of 20 candidate hub genes that were correlated with clinical subtypes were found in the blue module. In addition, we found ERBB2 and WNT11 that have not been studied in CC and verified their differential expression in CC through quantitative real-time PCR experiments. For the first time, we have described the transcriptome characteristics of CC. These results suggest that cystic CC and fusiform CC have different molecular mechanisms. The bi-omics-identified novel candidate genes and pathways might be helpful for personalized treatment and are of great clinical significance for CC.


Cobalt Superoxo and Alkylperoxo Complexes Derived from Reaction of Ring-Cleaving Dioxygenase Models with O2.
The syntheses and O2 reactivities of active-site models of cobalt-substituted ring-cleaving dioxygenases are presented. The pentacoordinate cobalt(II)-aminophenolate complex, [Co(TpMe2)(tBu2APH)], gives rise to two distinct dioxygen adducts at reduced temperatures. The first is a paramagnetic (S = 1/2) cobalt(III)-superoxo species that was characterized with spectroscopic and computational techniques. The identity of the second Co/O2 adduct was elucidated by X-ray crystallography, which revealed an unprecedented cobalt(III)-alkylperoxo structure generated by O2 addition to the metal ion and ligand. These results provide synthetic precedents for proposed intermediates in the catalytic cycles of O2-activating cobalt enzymes.


Saccharification of rice straw by cellulase from a local Trichoderma harzianum SNRS3 for biobutanol production.
Rice straw has shown to be a promising agricultural by-product in the bioconversion of biomass to value-added products. Hydrolysis of cellulose, a main constituent of lignocellulosic biomass, is a requirement for fermentable sugar production and its subsequent bioconversion to biofuels such as biobutanol. The high cost of commercial enzymes is a major impediment to the industrial application of cellulases. Therefore, the use of local microbial enzymes has been suggested. Trichoderma harzianum strains are potential CMCase and β-glucosidase producers. However, few researches have been reported on cellulase production by T. harzianum and the subsequent use of the crude cellulase for cellulose enzymatic hydrolysis. For cellulose hydrolysis to be efficiently performed, the presence of the whole set of cellulase components including exoglucanase, endoglucanase, and β-glucosidase at a considerable concentration is required. Biomass recalcitrance is also a bottleneck in the bioconversion of agricultural residues to value-added products. An effective pretreatment could be of central significance in the bioconversion of biomass to biofuels. Rice straw pretreated using various concentrations of NaOH was subjected to enzymatic hydrolysis. The saccharification of rice straw pretreated with 2% (w/v) NaOH using crude cellulase from local T. harzianum SNRS3 resulted in the production of 29.87 g/L reducing sugar and a yield of 0.6 g/g substrate. The use of rice straw hydrolysate as carbon source for biobutanol fermentation by Clostridium acetobutylicum ATCC 824 resulted in an ABE yield, ABE productivity, and biobutanol yield of 0.27 g/g glucose, 0.04 g/L/h and 0.16 g/g glucose, respectively. As a potential β-glucosidase producer, T. harzianum SNRS3 used in this study was able to produce β-glucosidase at the activity of 173.71 U/g substrate. However, for cellulose hydrolysis to be efficient, Filter Paper Activity at a considerable concentration is also required to initiate the hydrolytic reaction. According to the results of our study, FPase is a major component of cellulose hydrolytic enzyme complex system and the reducing sugar rate-limiting enzyme. Our study revealed that rice straw hydrolysate served as a potential substrate for biobutanol production and FPase is a rate-limiting enzyme in saccharification.


The lived experience of relationships after major burn injury.
To explore and describe the meaning of relationships after major burn injury. A major burn injury may represent a threat to preburn appearance and level of functioning. Social resources and interaction are considered important for minimising the negative impacts on life after burn through all phases of care and rehabilitation. Yet, the subjective experiences of relating to others after burns have not been extensively explored. The study was performed by using a phenomenological approach. A purposive sample (n = 14) of patients who had experienced major burns were interviewed in average 14 (5-35) months postinjury. The interviews were analysed by the phenomenological method of Giorgi. The essence of the experience of relationships was constituted by other people re-anchoring the burn-injured persons to preburn life, being confirming of the new self, as well as being sensitive, competent and safeguarding with respect to actual and potential problems and harms. New bodily limitations demanded assistive others. Moreover, a worry about the overall burden on close family was typical. The supportive actions from others were mostly described as positive, but could also be experienced as challenging, and sometimes even unwanted and interfering with the struggle for regained freedom. Across variation, an increased awareness of the meaning of mutual interdependence was typical. Supportive relationships with family, friends and health professionals were important for the return to society, preburn activities and a meaningful life. A perspective that values the significance of social support from family, friends, health professionals and others who are important for the burn-injured person, and that does not underestimate challenges that may be involved, is recommended during all phases of burn care. Health professionals should also acknowledge the importance of relationships when developing structured educational programmes and organising peer support.


Genome-scale investigation of olfactory system spatial heterogeneity.
The early olfactory system is organized in parallel, with numerous, specialized subsystems established by the modular and topographic projections of sensory inputs. While these anatomical sub-systems are in many cases demarcated by well-known marker genes, we stand to learn considerably more about their possible functional specializations from comprehensive, genome-scale descriptions of their molecular anatomy. Here, we leverage the resources of the Allen Brain Atlas (ABA)-a spatially registered compendium of gene expression for the mouse brain-to investigate the early olfactory system's genomic anatomy. We cluster thousands of genes across thousands of voxels in the ABA to derive several novel parcellations of the olfactory system, and concomitantly discover novel sets of enriched, subregion-specific genes that can serve as a starting point for future inquiry.


Autoantibody response and pregnancy-related pathology induced by combined LPS and tetanus toxoid hyperimmunization in BALB/c and C57BL/6 mice.
Recent data concerning antiphospholipid syndrome (APS) induction have shown that β2-glycoprotein I (β2GPI) binds lipopolysaccharide (LPS), which results in conformational changes, exposition of a cryptic epitope and possible pathological anti-β2GPI antibody production. In order to investigate the effects of LPS on the induction of APS-related pathology, we performed hyperimmunization of BALB/c and C57BL/6 mice with LPS, alone or in combination with tetanus toxoid (TTd), a protein structurally similar to β2GPI. We report that, although high affinity pathological anti-β2GPI antibodies were produced in all groups of animals, the reproductive pathology was recorded only in mice that received both LPS and TTd, implying on the important roles of both infections and molecular mimicry in APS pathogenesis. Moreover, APS-related reproductive pathology was more pronounced in BALB/c (lowered fertility and fecundity) than C57BL/6 mice (lowered fecundity), which correlated well with the disruption in natural antibody network observed in BALB/c mouse strain.


Glutamates 99 and 107 in transmembrane helix III of subunit I of cytochrome bd are critical for binding of the heme b595-d binuclear center and enzyme activity.
Cytochrome bd is a quinol oxidase of Escherichia coli under microaerophilic growth conditions. Coupling of the release of protons to the periplasm by quinol oxidation to the uptake of protons from the cytoplasm for dioxygen reduction generates a proton motive force. On the basis of sequence analysis, glutamates 99 and 107 conserved in transmembrane helix III of subunit I have been proposed to convey protons from the cytoplasm to heme d at the periplasmic side. To probe a putative proton channel present in subunit I of E. coli cytochrome bd, we substituted a total of 10 hydrophilic residues and two glycines conserved in helices I and III-V and examined effects of amino acid substitutions on the oxidase activity and bound hemes. We found that Ala or Leu mutants of Arg9 and Thr15 in helix I, Gly93 and Gly100 in helix III, and Ser190 and Thr194 in helix V exhibited the wild-type phenotypes, while Ala and Gln mutants of His126 in helix IV retained all hemes but partially lost the activity. In contrast, substitutions of Thr26 in helix I, Glu99 and Glu107 in helix III, Ser140 in helix IV, and Thr187 in helix V resulted in the concomitant loss of bound heme b558 (T187L) or b595-d (T26L, E99L/A/D, E107L/A/D, and S140A) and the activity. Glu99 and Glu107 mutants except E107L completely lost the heme b595-d center, as reported for heme b595 ligand (His19) mutants. On the basis of this study and previous studies, we propose arrangement of transmembrane helices in subunit I, which may explain possible roles of conserved hydrophilic residues within the membrane.


Multi-Geometry Parameters Optimization of Large-Area Roll-to-Roll Nanoimprint Module Using Grey Relational Analysis and Artificial Neural Network.
Micro- and nanofabrication on polymer substrate is integral to the development of flexible electronic devices, including touch screens, transparent conductive electrodes, organic photovoltaics, batteries, and wearable devices. The demand for flexible and wearable devices has spurred interest in large-area, high-throughput production methods. Roll-to-roll (R2R) nanoimprint lithography (NIL) is a promising technique for producing nano-scale patterns rapidly and continuously. However, bending in a large-scale R2R system can result in non-uniform force distribution during the imprinting process, which reduces pattern quality. This study investigates the effects of R2R imprinting module geometry parameters on force distribution via simulation, using grey relational analysis to identify optimal parameter levels and ANOVA to determine the percentage of each parameter contribution. The study also investigates the length and force ratio on a backup roller used for bending compensation. The simulation results and the artificial neural network (ANN) model enable the prediction of nip pressure and force distribution non-uniformity along the roller, allowing the selection of the optimal roller geometry and force ratio for minimal non-uniformity on a specific R2R system. An experiment was conducted to validate the simulation results and ANN model.


Pseudoaneurysm of the brachiocephalic arteries: a complication of percutaneous internal jugular vein catheterization.
Three cases of arterial pseudoaneurysm formation as a complication of percutaneous internal jugular vein catheterization are presented. Laceration of the posterior common carotid artery or thyrocervical trunk was the source for these lesions. The common antecedent event was the attempted cannulation of the internal jugular vein with the lateral approach. A transcervical rather than transthoracic approach for treatment of these lesions was selected because of its low morbidity rate. This report documents another complication of internal jugular vein cannulation and recommends methods for prevention and successful treatment should it occur.


Consequences of living environment insecurity on health and well-being in southwest China: The role of community cohesion and social support.
Growing populations in developing countries have exacerbated inequality in the spatial distribution of living environments. As a result, whether living environment factors matter to health and well-being is increasingly attracting policy and scholarly attention. Yet, crucial knowledge gaps remain regarding the implications, consequences and mechanisms of one's living environment on health and well-being in developing countries. This study examined the association between living environment and psychological distress, self-rated health and satisfaction with life among Chinese adults. Furthermore, it also explored the moderating role of community cohesion and the mediating role of social support on these factors. Using probability proportionate to size sampling methods, 3765 respondents and 148 community organisers were recruited from 160 communities in Yunnan, China, in 2018. Ordinary least squares regression and structural equation modelling were conducted. The findings indicate that residents who lived in communities with higher levels of environment insecurity had lower levels of subjective health and well-being (psychological distress: β = 1.088, p < 0.001; self-rated health: β = -0.104, p < 0.01; satisfaction with life: β = -0.164, p < 0.001). Moreover, community cohesion played a moderating role in the relationship between living environment insecurity and self-rated health (β = 0.212, p < 0.05) and satisfaction with life (β = 0.183, p < 0.05); however, it had no significant effects on psychological distress. Furthermore, the moderated effects of community cohesion on living environment insecurity were mediated through perceived social support for self-rated health (proportion of total effect mediated = 11%) and satisfaction with life (proportion of total effect mediated = 29%). The present findings have implications for policy makers and community committees who can work towards social inequality in developing countries. The integrated programmes of improving living conditions and strengthening community capacity are crucial to residents' health and well-being.


Crosstalk within a functional INO80 complex dimer regulates nucleosome sliding.
Several chromatin remodellers have the ability to space nucleosomes on DNA. For ISWI remodellers, this involves an interplay between H4 histone tails, the AutoN and NegC motifs of the motor domains that together regulate ATPase activity and sense the length of DNA flanking the nucleosome. By contrast, the INO80 complex also spaces nucleosomes but is not regulated by H4 tails and lacks the AutoN and NegC motifs. Instead nucleosome sliding requires cooperativity between two INO80 complexes that monitor DNA length simultaneously on either side of the nucleosome during sliding. The C-terminal domain of the human Ino80 subunit (Ino80CTD) binds cooperatively to DNA and dimerisation of these domains provides crosstalk between complexes. ATPase activity, rather than being regulated, instead gradually becomes uncoupled as nucleosome sliding reaches an end point and this is controlled by the Ino80CTD. A single active ATPase motor within the dimer is sufficient for sliding.


[Significance of radiotherapy for the treatment of primary bone and soft tissue sarcomas].
Recent data from the literature as well as our own treatment results of soft tissue sarcomas and primary bone tumours with the exception of osteosarcomas indicate, that: 1. A conservative, limb sparing therapy should be attempted in peripheral bone tumours and soft tissue sarcomas. This may be achieved in 85% to 97%. 2. The recurrence rate is in a range of 3% to 22%, which does not exceed the results of ablative surgery. 80% of possible recurrences can be controlled by consecutive surgery. 3. After the development of improved treatment techniques, the complication rate is less than 10%. 4. Conservative limb sparing treatment does not decrease the survival rate. However, these results are not derived from randomised studies. 5. Apart from osteosarcomas and Ewing sarcomas adjuvant chemotherapy is recommended for G3 tumours. The time interval for evidence of metastases may be doubled by this treatment modality. 6. There is some evidence that neutron-boost therapy may improve local control rates in T2 and T3 soft tissue sarcomas.


Managing antiretroviral-associated liver disease.
The use of all potent drugs is associated with toxicities and antiretroviral (ARV) drugs are no exception. Antiretroviral therapy-associated hepatic toxicity is of increasing concern in the management of patients with HIV/AIDS. Liver toxicity has been reported in some HIV-infected patients being treated with drugs from all of these classes of ARV drugs: protease inhibitors (PIs), nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). Although the majority of cases involve asymptomatic elevations of liver enzymes (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]), severe, and, in a minority of cases, life threatening, liver disease has been reported in patients treated with ARV drugs. The exact causes of elevated plasma levels of AST and ALT are complex and, in many cases, obscure. The combination of viral hepatic disease, drugs that act adversely directly on the liver and drugs that act on other systems of the body which in turn, adversely affect the liver, can result in hepatic toxicity. Such toxicity may be inappropriately attributed solely to the direct effect of a drug. Knowledge of the possible causes of liver toxicity, and ways to avoid it, should reduce the risk of developing hepatotoxicity. The physician's task is to prevent the development of liver toxicity, e.g., by choosing appropriate therapeutic regimens and by careful management of the patient. This involves frequent monitoring of the patient, both clinically and by utilizing liver function tests on a regular basis. If signs and symptoms of liver disease do develop, prompt and expert management is essential. This review discusses the influence of a number of factors on hepatic toxicity including viral hepatitis, insulin resistance and the specific ARV drugs used in the treatment of patients with HIV/AIDS.


Nonblocking 3 × 3 polymer thermo-optic switch array based on total-internal-reflection effect.
A nonblocking 3×3 polymer thermo-optic (TO) switch array based on the total-internal-reflection (TIR) effect is demonstrated. The switch array consists of three 2×2 TIR TO switch elements with a total device length of 15.5 mm, and achieves nine switching states with no more than one switch element working. The fabricated 3×3 TO switch array shows excellent switching performance with low crosstalk for all switching states of less than -20 dB. The power consumption of each switch element is below 53 mW, and the switching rise time and fall time are ∼450 and 550 μs at 1550 nm wavelength, respectively.


Synthesis of metal anthranilate complexes: catalytic and antipathogenic studies.
Anthranilic acid is an active compound with diverse biological activities such as anti-inflammatory, antineoplastic, anti-malarial and α-glucosidase inhibitory properties. It can also chelate transition metals to form complexes with applications as antipathogens, photoluminescent materials, corrosion inhibitors, and catalysts. Anthranilic acid complexes (1-10) of Zn(II), Bi(III), Ag(I), Fe(II), Co(II), Cu(II), Mn(II), Al, Ni(II), and Cr(III) were synthesized and characterized using thermogravimetric (TGA), elemental analysis, FT-IR, UV-vis spectrometry, mass spectrometry and magnetic susceptibility. The morphology and size of metal complex (1-10) particles were determined by scanning electron microscope (SEM) and the surface area was determined by BET analysis. TGA and CHN analysis data indicated that the stoichiometries of complexes were 1:2 metal/ligand except for Ag(I), Al and Bi. Furthermore, DFT study was performed to optimize the structure of selected complexes. The complexes (1-10) were evaluated for their catalytic activity in the reduction of 4-nitrophenol (4-NP), antibacterial activity against S. aureus, P. aeroginosa and E. coli as well as their antifungal activity against F. solani and A. niger. The complexes were also tested against the second-stage juveniles (J2) root-knot nematodes. Co(II) complex 5 and Cu(II) complex 6 showed high catalytic activity for the reduction of 4-NP to 4-aminophenol (4-AP). Ag(I) complex 3 showed the best activity against the pathogens that were tested namely clinically important bacteria S. aureus, P. aeroginosa and E. coli, commercially important fungi F. solani and A. niger and J2 root-knot nematodes M. javanica.


Depot-specific differences in fat mass expansion in WT and ob/ob mice.
The study was designed to investigate the cellular mechanisms underlying the differential fat expansion in different fat depots in wild type (WT) and ob/ob (OB) mice. At 6 weeks old, no differences in fat mass were found between SAT and VAT in WT mice, while O-SAT showed significantly higher weight than that of O-VAT. The average adipocyte size of SAT (~ 4133.47 μm2) was smaller than that of VAT (~ 7438.91 μm2) in OB mice. O-SAT preadipocytes gained higher triglyceride contents and higher levels of PPARγ and C/EBPα than did O-VAT preadipocytes upon in vitro differentiation. W-SAT and W-VAT displayed no significant differences in fatty acid uptake, while 1.36 fold significantly higher fatty acid uptake was found in O-SAT compared to O-VAT. Approximately 52% of the radioactivity recovered in cellular lipids was found in TAG in O-SAT, which was significantly higher than the other three adipocyte types. Significantly more radiolabelled oleic acid was β-oxidized to CO2 in adipocytes from O-VAT than that from O-SAT. ATP production was significantly lower in W-SAT compared with W-VAT, whereas no significantly ATP level was observed between O-SAT and O-VAT. Expression of UCP-1 in SAT from either WT or OB mice was significantly higher than the counterpart of VAT, which demonstrated higher uncoupled respiration and lower oxidative phosphorylation in SAT. Together, a combined increase in adipogenesis and FA uptake, and decreases in β-oxidation and ATP production, contributed to greater expansion of SAT compared to VAT in obese mice.


circ_0006089 promotes gastric cancer growth, metastasis, glycolysis, and angiogenesis by regulating miR-361-3p/TGFB1.
Circular RNA (circRNA) participates in a variety of pathophysiological processes, including the development of gastric cancer (GC). However, the role of circ_0006089 in GC progression and its underlying molecular mechanism need to be further revealed. Quantitative real-time PCR was utilized for detecting circ_0006089, microRNA (miR)-361-3p and transforming growth factor-β1 (TGFB1) expression. The interaction between miR-361-3p and circ_0006089 or TGFB1 was confirmed using a dual-luciferase reporter assay and an RNA immunoprecipitation (RIP) assay. Cell proliferation, metastasis, apoptosis, and angiogenesis were determined using colony formation assay, EdU assay, transwell assay, flow cytometry, and tube formation assay. Cell glycolysis was evaluated by detecting glucose consumption, lactate production, and ATP levels. In addition, western blot (WB) analysis was used to measure protein expression. Xenograft tumor models were used to assess the effect of circ_0006089 knockdown on GC tumorigenesis. circ_0006089 had been found to be upregulated in GC tissues and cells, and it could act as an miR-361-3p sponge. circ_0006089 knockdown suppressed GC proliferation, metastasis, glycolysis, angiogenesis, and increased apoptosis, while this effect could be revoked by miR-361-3p inhibitor. TGFB1 was targeted by miR-361-3p, and its overexpression reversed the effects of miR-361-3p on GC cell function. Also, circ_0006089 promoted TGFB1 expression via sponging miR-361-3p. Animal experiments showed that silenced circ_0006089 inhibited GC tumorigenesis through the miR-361-3p/TGFB1 pathway. Our results revealed that the circ_0006089/miR-361-3p/TGFB1 axis contributed to GC progression, confirming that circ_0006089 might be a potential therapeutic target for GC.


Microplastics in intertidal water of South Australia and the mussel Mytilus spp.; the contrasting effect of population on concentration.
Microplastics, plastic particles <5 mm in size, are of global concern as human-caused pollutants in marine and fresh waters, and yet little is known of their distribution, behaviour and ecological impact in the intertidal environment of South Australia. This study confirms for the first time, the presence of microplastic in the South Australian intertidal ecosystem by quantifying the abundance of particles in intertidal water and in the keystone species, the blue mussel, Mytilus spp., an important fisheries species, at ten and six locations respectively, along the South Australian coastline. For a remote region known for its pristine environment, microplastic concentration in intertidal water was found to be low to moderate (mean = 8.21 particles l-1 ± 4.91) relative to global levels and microplastic abundance in mussels (mean = 3.58 ± 8.18 particles individual-1) was within the range also reported globally. Microplastic particles were ubiquitous across sites and bioavailable by size in water (mean = 906.36 μm) and in mussel (mean = 983.29 μm) raising concerns for the health of South Australia's unique coastal ecosystems and for the human food chain. Furthermore, a positive correlation was found between human coastal population size and microplastic concentration in intertidal water, irrespective of influences from industry - tourism, fishing and shipping ports. FTIR analysis determined plastic type to include polyamide (PA), polyethylene (PE), polypropylene (PP), acrylic resin, polyethyleneterephthalate (PET) and cellulose, suggesting synthetic and semi-synthetic particles from single-use, short-life cycle products, fabrics, ropes and cordage. Our findings shed light on the urgent need to establish the local sources of microplastic pollution in order to assist the community, industry and government to reduce the impact of microplastic on the fragile marine systems within South Australian intertidal waters and on the organisms associated with the human food chain.


Purification and characterization of a novel bacteriocin produced by Enterococcus faecalis strain RJ-11.
Lactic acid bacteria exhibiting activity against the gram-positive bacterium Bacillus subtilis were isolated from rice bran. One of the isolates, identified as Enterococcus faecalis RJ-11, exhibited a wide spectrum of growth inhibition with various gram-positive bacteria. A bacteriocin purified from culture fluid, designated enterocin RJ-11, was heat stable and was not sensitive to acid and alkaline conditions, but it was sensitive to several proteolytic enzymes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that enterocin RJ-11 had a molecular weight of 5,000 in its monomeric form. The amino acid sequence determined for purified enterocin RJ-11 exhibited high levels of similarity to the sequences of enterocins produced by Enterococcus faecium.


Actinobacillus pleuropneumoniae serotype 1 carrying the defined aroA mutation is fully avirulent in the pig.
The aroA gene from Actinobacillus pleuropneumoniae serotype 1 reference strain 4074 was isolated and sequenced. The gene complemented the aroA mutation in Escherichia coli AB2829. A kanamycin resistance cassette was inserted into the aroA gene and the mutant gene was reintroduced into A. pleuropneumoniae by allelic replacement. Intratracheal infection of susceptible pigs with A. pleuropneumoniae aroA caused no signs of respiratory disease or lung lesions in any of the animals at a dose 10(4) times the dose reliably known to induce acute pleuropneumonia; all animals infected with the unaltered control strain developed acute disease. The aroA mutant was rapidly eliminated from the lungs and tonsil of infected animals. The mutant may represent a safely attenuated strain for use in live bacterial vaccination or the delivery of antigen by the intranasal route. However, the residence time of the mutant in the respiratory tract of the pig may be too short for it to be useful in generating a protective immune response.


Object permanence after a 24-hr delay and leaving the locale of disappearance: the role of memory, space, and identity.
Fourteen-month-old infants saw an object hidden inside a container and were removed from the disappearance locale for 24 hr. Upon their return, they searched correctly for the hidden object, demonstrating object permanence and long-term memory. Control infants who saw no disappearance did not search. In Experiment 2, infants returned to see the container either in the same or a different room. Performance by room-change infants dropped to baseline levels, suggesting that infant search for hidden objects is guided by numerical identity. Infants seek the individual object that disappeared, which exists in its original location, not in a different room. A new behavior, identity-verifying search, was discovered and quantified. Implications are drawn for memory, spatial understanding, object permanence, and object identity.


Formation and Reactivity of a C-P-N-Sc Four-Membered Ring: H2 , O2 , CO, Phenylsilane, and Pinacolborane Activation.
Reaction of scandium dimethyl complex LScMe2 (L=[2-{N(DIPP)C(Ph)N}C6 H4 CH=N(DIPP)]- , DIPP=2,6-(iPr)2 C6 H3 ) (2) with bulky phosphine ArPH2 (Ar=2,6-{3,5-(Me)2 C6 H3 }2 C6 H3 ) gives an unprecedented scandium 2,3-azaphosphametallacyclobutane complex L'Sc (L'=[2-{N(DIPP)C(Ph)N}C6 H4 C(H)P(2,6-{3,5-(CH3 )2 C6 H3 }2 C6 H3 )N(DIPP)]3- ) (3) that contains a C-P-N-Sc four-membered ring. Complex 3 was characterized by X-ray crystallography, revealing a short distance between the Sc and P atoms (2.786(1) Å), but without a direct Sc-P bonding interaction. The formation of 3 involves a complete cleavage of the imino bond of the tridentate ligand L and C-P and N-P bond coupling. Complex 3 not only activates H2 under mild conditions to give a H-H bond cleavage product, but also reacts with O2 , CO, phenylsilane, and pinacolborane to produce novel products through cleavage and formation of various bonds.


Duration of breast-feeding in Bangladesh.
The duration of exclusive breast-feeding in Bangladesh is low. Though several studies have been carried out on breast-feeding in Bangladesh, the factors influencing the duration of breast-feeding are not studied. The purpose of this study was to assess the duration of breast-feeding among children in Bangladesh and to study socio-economic and demographic factors affecting the duration. The study included 5068 mother-child pairs, selected on the basis of Bangladesh Demographic Health Survey (BDHS) of 1999-2000. The risk of giving supplementary food at different ages was estimated by life table survival method. Independent effects of total duration of breast-feeding of each of the study variables were estimated by using Cox's regression model. The median duration of full breast-feeding was 3.67 months whereas mean and median durations of total breast-feeding was 31.3 and 30 months respectively. Life table analysis showed that 69.9 per cent women gave supplementary food to their babies before reaching six months of age. Cox's regression analysis revealed that women who had lived in rural areas were less likely to terminate breast-feeding than those living in urban areas. Women who had completed at least secondary education were more likely to stop breast-feeding than less or uneducated mothers. Children born in high economic status families had higher risk of stopping breast-feeding compared to those in low economic status families. Further, the deliveries assisted by the relatives had lower risk of terminating breastfeeding than by the health professionals. According to the study results, women with higher education, high economic level, lower birth interval and delivery assisted by health personnel had lower duration of breastfeeding. Future breast-feeding programme in Bangladesh should give special attention to these women since they breast-feed relatively shorter periods of time.


Distributed representation of vocalization pitch in marmoset primary auditory cortex.
The pitch of vocalizations is a key communication feature aiding recognition of individuals and separating sound sources in complex acoustic environments. The neural representation of the pitch of periodic sounds is well defined. However, many natural sounds, like complex vocalizations, contain rich, aperiodic or not strictly periodic frequency content and/or include high-frequency components, but still evoke a strong sense of pitch. Indeed, such sounds are the rule, not the exception but the cortical mechanisms for encoding pitch of such sounds are unknown. We investigated how neurons in the high-frequency representation of primary auditory cortex (A1) of marmosets encoded changes in pitch of four natural vocalizations, two centred around a dominant frequency similar to the neuron's best sensitivity and two around a much lower dominant frequency. Pitch was varied over a fine range that can be used by marmosets to differentiate individuals. The responses of most high-frequency A1 neurons were sensitive to pitch changes in all four vocalizations, with a smaller proportion of the neurons showing pitch-insensitive responses. Classically defined excitatory drive, from the neuron's monaural frequency response area, predicted responses to changes in vocalization pitch in <30% of neurons suggesting most pitch tuning observed is not simple frequency-level response. Moreover, 39% of A1 neurons showed call-invariant tuning of pitch. These results suggest that distributed activity across A1 can represent the pitch of natural sounds over a fine, functionally relevant range, and exhibits pitch tuning for vocalizations within and outside the classical neural tuning area.


Identification and localization of neurohypophysial peptides in the brain of a caecilian amphibian, Typhlonectes natans (Amphibia: Gymnophiona).
The amphibian order Gymnophiona contains more than 150 different species of caecilians. The characterization and distribution of neurohypophysial peptides, however, has not been described for any member of this order. By using high-performance liquid chromatography, radioimmunoassay, and mass spectrometry, we identified the peptide arginine vasotocin (AVT) in brain and pituitary extracts from the caecilian Typhlonectes natans. By using immunocytochemistry, we found five populations of AVT-immunoreactive (AVT-ir) cells in the brain of T. natans. AVT-ir cell bodies were located in the preoptic area, amygdala pars medialis, ventral thalamus, dorsal hypothalamic nucleus, and nucleus of the solitary tract. AVT-ir fibers and terminal fields were widespread. We also identified a mesotocin-like peptide. The distribution of this peptide in the brain of T. natans was more restricted than the distribution of AVT. Mesotocin-like-immunoreactive cell bodies were located almost exclusively in the preoptic area, with only a few other cells located in the amygdala pars medialis. This caecilian species, therefore, possesses neurohypophysial peptides that are similar in their structure and distribution to the peptides found in anuran and urodele amphibian orders.


Purification, crystallization, and properties of F1-ATPase complexes from the thermoalkaliphilic Bacillus sp. strain TA2.A1.
Recently, we reported the cloning of the atp operon encoding for the F(1)F(0)-ATP synthase from the extremely thermoalkaliphilic bacterium Bacillus sp. strain TA2.A1. In this study, the genes encoding the F(1) moiety of the enzyme complex were cloned from the atp operon into the vector pTrc99A and expressed in Escherichia coli in two variant complexes, F(1)-wt consisting of subunits alpha(3)beta(3)gammadeltaepsilon and F(1)Deltadelta lacking the entire delta-subunit as a prerequisite for overproduction and crystallization trials. Both F(1)-wt and F(1)Deltadelta were successfully overproduced in E. coli and purified in high yield and purity. F(1)Deltadelta was crystallized by micro-batch screening yielding three-dimensional crystals that diffracted to a resolution of 3.1A using a synchrotron radiation source. After establishing cryo and dehydrating conditions, a complete set of diffraction data was collected from a single crystal. No crystals were obtained with F(1)-wt. Data processing of diffraction patterns showed that F(1)Deltadelta crystals belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell parameters of a=121.70, b=174.80, and c=223.50A, alpha, beta, gamma=90.000. The asymmetric unit contained one molecule of bacterial F(1)Deltadelta with a corresponding volume per protein weight (V(M)) of 3.25A(3) Da(-1) and a solvent content of 62.1%. Silver staining of single crystals of F(1)Deltadelta analyzed by SDS-PAGE revealed four bands alpha, beta, gamma, and epsilon with identical M(r)-values as those found in the native F(1)F(0)-ATP synthase isolated from strain TA2.A1 membranes. ATPase assays of F(1)Deltadelta crystals exhibited latent ATP hydrolytic activity that was highly stimulated by lauryldimethylamine oxide, a hallmark of the native enzyme.


The potential role of ubiquitination and deubiquitination in melanogenesis.
Melanogenesis is a critical biochemical process in which melanocytes produce melanin, a crucial element involved in the formation of coat colour in mammals. According to several earlier studies, melanocytes' post-translational modifications of proteins primarily control melanogenesis. Among the many post-translational changes that can affect melanin production, ubiquitination and deubiquitination can keep melanin production going by changing how proteins that are related to melanin are broken down or kept stable. Ubiquitination and deubiquitination maintain ubiquitin homeostasis, which is a highly dynamic process in balance under the action of E3 ubiquitin ligase and deubiquitinating enzymes. However, the regulatory mechanisms underlying ubiquitination and deubiquitination in melanogenesis are yet to be thoroughly investigated. As a result, there has been a growing focus on exploring the potential correlation between melanogenesis, ubiquitination and deubiquitination. This study discusses the mechanisms of ubiquitination and deubiquitination in the context of melanogenesis, a crucial process for enhancing mammalian coat coloration and addressing pigment-related diseases.


ABCG2 Overexpression Confers Poor Outcomes in Hepatocellular Carcinoma of Elderly Patients.
Backgroung/Aim: Breast cancer resistance protein [BCRP/ATP-binding cassette subfamily G member 2 (ABCG2)] is a member of the ATP-binding cassette transporter family, used as a maker of cancer stem cells (CSCs) and thought to be responsible for drug resistance by pumping them out of cells. However, its precise role in various cancer types has been controversial, and the aim of this study was to investigate the expression of ABCG2 in hepacellular carcinoma (HCC) and relate the results to established prognostic factors. We conducted analysis of 181 HCC and paired-match adjacent normal liver tissue by immunohistochemistry from tissue array of slides. The mean score for ABCG2 expression was higher in tumor than in adjacent normal liver tissue of HCC patients (p<0.001). There was a statistically significant correlation between ABCG2 expression and age, differentiation status and hepatitis B surface antigen (p=0.031, p=0.015 and p=0.033, respectively). Additionally, increased expression of ABCG2 in HCC and its statistically significant correlation with hepatitis B surface antigen was found in elderly (p=0.039), not in younger patients (p=0.518). Importantly, by using Kaplan-Meier and Cox regression analysis, overall survival in patients with high expression of ABCG2 was found reduced in elderly patients (p=0.029 and p=0.081, respectively). ABCG2 can be used as a target for the development of novel therapies in HCC.


Larger ATV engine size correlates with an increased rate of traumatic brain injury.
Since the introduction of all-terrain vehicles (ATV) to the United States in 1971, injuries and mortalities related to their use have increased significantly. Furthermore, these vehicles have become larger and more powerful. As there are no helmet requirements or limitations on engine-size in the State of Alabama, we hypothesised that larger engine size would correlate with an increased incidence of traumatic brain injury (TBI) in patients following an ATV crash. Patient and ATV data were prospectively collected on all ATV crashes presenting to a level one trauma centre from September 2010 to May 2013. Collected data included: demographics, age of driver, ATV engine size, presence of helmet, injuries, and outcomes. The data were grouped according to the ATV engine size in cubic centimetres (cc). For the purposes of this study, TBI was defined as any type of intracranial haemorrhage on the initial computed tomography scan. There were 61 patients identified during the study period. Two patients (3%) were wearing a helmet at the time of injury. Patients on an ATV with an engine size of 350 cc or greater had higher Injury Severity Scores (13.9 vs. 7.5, p ≤ 0.05) and an increased incidence of TBI (26% vs. 0%, p ≤ 0.05) when compared to patients on ATV's with an engine size less than 350 cc. Patients on an ATV with an engine size of 350 cc or greater were more likely to have a TBI. The use of a helmet was rarely present in this cohort. Legislative efforts to implement rider protection laws for ATVs are warranted.


Characterization of the membrane ion currents of a model molluscan muscle, the accessory radula closer muscle of Aplysia californica. III. Depolarization-activated Ca current.
1. The accessory radula closer (ARC) muscle of Aplysia californica and its innervation is a model preparation for the study of the neural and cellular mechanisms of behavioral plasticity. Much of the plasticity is due to modulation of contractions of the muscle by a variety of neurotransmitters and peptide cotransmitters. Preliminary to investigating the cellular mechanisms of this modulation, we have characterized the major membrane ion currents present in the unmodulated ARC muscle and their likely roles in normal contraction. We have studied single dissociated but functionally intact ARC muscle fibers under voltage clamp. This is the last of three papers describing this work. In the first paper we characterized two currents prominent at hyperpolarized voltages, a classical inwardly rectifying K current and a Cl current induced by elevated intracellular Cl-. In the second paper we examined two large outward K currents activated at more depolarized voltages, an "A" current and a delayed rectifier. 2. In this paper, we describe an inward depolarization-activated Ca current that underlies and is normally completely masked by the K currents and is revealed when they are blocked. 3. The Ca current begins to activate above -40 or -30 mV. It is fully available for activation at voltages more negative than -60 mV. It activates in milliseconds, then inactivates relatively slowly with maintained depolarization. The current is larger and inactivates slower when it is carried by Ba2+ rather than Ca2+. The inactivation is current rather than voltage dependent. The current is blocked by Co2+, Cd2+, and with a characteristic time dependence, by the dihydropyridine Ca-channel antagonist nifedipine. 4. These properties of the current characterize it as a high-threshold, L-type Ca current. 5. This current most likely provides Ca2+ necessary for contraction of the ARC muscle.


[Effect of Electroacupuncture Stimulation of Heart and Lung Meridians on Expression of Myocardial HCN 2 in Acute Myocardial Ischemia Rats].
To observe the effect of electroacupuncture (EA) stimulation of Heart and Lung meridians on the expression of hyperpolarization-activated cyclic nucleotide gated potassium channel 2 (HCN 2) mRNA and protein in the myocar-dium of the left ventricle in acute myocardial ischemia (AMI) rats, so as to underlying its mechanism in improving ischemic myocardial injury. A total of 120 male SD rats were randomly divided into normal control, model, EA-Heart Meridian (HT) and EA-Lung Meridian (LU) groups (n=30 rats in each group). The AMI model was established by occlusion of the anterior descending branch of coronary artery. EA (1 mA, 10 Hz) was applied to bilateral "Shenmen" (HT 7) or "Taiyuan" (LU 9) and the spot about 1 mm apart from the HT 7 or LU 9 on the axopetal end for 15 min on the second day after modeling, once a day for 7 days. The left ventricular myocardium tissue was sampled for determining the expression levels of HCN 2 mRNA and protein with qPCR and Western blot, separately. After AMI, both HCN 2 mRNA and protein expression levels in the model group were significantly decreased in comparison with the normal control group (P<0.01). Following EA intervention, the expression levels of HCN 2 mRNA and protein expressions were significantly up-regulated relevant to the model group (P<0.01).The effects were more obvious in the EA-HT group than in the EA-LU group, and the expression levels were higher in the left ventricular myocardium tissue sampled on the next day after the treatment than those in the tissue sampled immediately after the treatment (P<0.01). Both EA stimulation of Heart Meridian and Lung Meridian can improve the expression of HCN 2 mRNA and protein in the myocardium in AMI rats, which existing a delayed effect and meridian specificity.


Multiparametric ultrasound in liver diseases: an overview for the practising clinician.
Ultrasound (US) is usually the first and most commonly used tool in the diagnostic algorithm for liver disease. It is widely available, non-invasive and offers a real-time assessment of the liver in several anatomic planes, using different US modalities such as greyscale imaging, Doppler, elastography and contrast-enhanced US. This multiparametric ultrasound (MPUS) provides more information of the examined structures and allows for a faster and more accurate diagnosis, usually at the point of care, thus reducing the requirement for some invasive and more expensive methods. Current data on the MPUS in hepatology are summarised in this review, mostly focused on its use for non-invasive staging of liver fibrosis, detection and classification of portal hypertension and oesophageal varices, prognosis in chronic liver diseases and characterisation of focal liver lesions (FLLs). Based on the available data, we propose practical algorithms for clinical use of MPUS in chronic liver disease and FLL.