bluuebunny/crossref_metadata_2025 huggingface dataset:
Precipitating causes and outcomes of chronic obstructive pulmonary disease exacerbation at a tertiary care center in northeast Thailand
Authors: Reechaipichitkul Wipa
Published: August 2015

Background: Acute exacerbation of chronic obstructive pulmonary disease (COPD) is a leading cause of hospitalization and economic burden. Frequent exacerbations impair quality of life and effect decline of lung function.

Objective: We evaluated characteristics of COPD patients with frequent exacerbations. The precipitating causes, outcomes, hospital stay, and cost of admission were also determined.

Methods: The study population included COPD patients admitted because of acute COPD exacerbation at Srinagarind Hospital between 1 January 2006 and 31 December 2010.

Results: Over the 5-year period, 183 patients were admitted. Their mean age was 74.9 (SD 9.28) years and the male to female ratio was 170:13. Most patients (144; 79%) had one exacerbation per year and 39 (21%) had more than one per year. The group with more exacerbations, had a higher stage of the disease than those with only one exacerbation (p = 0.023), but there was no significant difference in the mortality rate (18% vs 14%, p = 0.53). A total of 245 episodes of acute exacerbation of COPD occurred in 183 patients. The mean duration of symptoms was 4.1 (SD 3.46) days. Forty-seven percent presented with Anthonisen type III, 42.4% with Anthonisen type II, and 10.6% with Anthonisen type I exacerbations. For 44 exacerbations (18%), the precipitating causes were not determined. The most common precipitating cause was pneumonia, which occurred in 90 episodes (36.7%). The second common was bronchitis (27.8%); followed by heart failure (8.2%), infected bronchiectasis (5.3%), diarrhea (1.2%), acute urinary retention (0.8%), unstable angina (0.4%), pneumothorax (0.4%), urinary tract infection (0.4%), atrial fibrillation (0.4%) and drug induced cough (0.4%). The organisms responsible for respiratory tract infection were identified in 31% cases of pneumonia and 18% of bronchitis cases. The top three common pathogens for pneumonia were Pseudomonas aeruginosa (9%), Acinetobacter baumannii (8%), and Klebsiella pneumoniae (8%). The top three common pathogens for bronchitis were P aeruginosa (7%), Haemophilus influenza (6%), and K pneumoniae (4%). About one quarter (25.3%) of acute exacerbations were complicated by respiratory failure. The mean duration of admission was 17.3 days (range 1-682 days). The mean cost of admission per exacerbation was 80,010 Thai baht (US $2,666) (range, 2,779-3,433,108 baht). The total cost for 245 exacerbations was 19.6 million baht ($653,000).

Conclusion: Respiratory tract infections were common causes of COPD exacerbation and one quarter of which developed respiratory failure. Preventive measures such as vaccination, smoking cessation, lung rehabilitation, and appropriate drug use are helpful.
Bioequivalence study of cefepime intramuscular injection
Authors: Jullangkoon Monchana , Jaruratanasirikul Sutep , Aeinlang Nanchanit
Published: August 2015

Background: Cefepime, a fourth generation cephalosporin, is a broad spectrum antibiotic effective against both Gram positive and Gram negative bacteria. It is available from several pharmaceutical firms in southeast Asia. We studied bioequivalence of two products.

Objective: To assess the bioequivalence of two cefepime formulations: Cefamax 1 g intramuscular (Siam Pharmaceutical, Bangkok, Thailand) as the test formulation and Maxipime 1 g (Bristol-Myers Squibb, Bangkok, Thailand) as the reference formulation.

Methods: The study was conducted as an open, randomized, two-period crossover trial with a 1 week washout period in 18 healthy volunteers. Each subject received a single dose of 1 g intramuscular injection of the test or reference formulation. Blood samples were collected via an intravascular catheter at several time points over a 12 h period. Plasma cefepime concentrations were quantified by HPLC with photodiode array detection at 280 nm. The statistical comparisons for pharmacokinetic parameters were made using a paired t test.

Results: There was no significant difference in the logarithmically transformed values of Cmax, AUC0-12, and AUC0-∞ between Cefamax and Maxipime using an analysis of variance (ANOVA). The 90% confidence intervals (CIs) of Cmax, AUC0-12, and AUC0-∞ between the two formulations were 91.17-105.78, 90.29-97.63, and 88.89-96.57, respectively. All subjects had good tolerance and no serious adverse events were observed.

Conclusion: Cefamax 1 g intramuscular formulation is bioequivalent to Maxipime 1 g intramuscular formulation based on 90% CIs for Cmax, AUC0-12, and AUC0-∞ within 80%-125%.
Surveillance of marine fish for ciguatera toxin at fish markets in Bangkok, Thailand
Authors: Sintunawa Chirapol , Wacharapluesadee Supaporn , Saraya Abhinbhen , Hemachudha Thiravat
Published: August 2015

Background: Ciguatera intoxication is a form of food poisoning. It is caused by the consumption of certain warm-water fish that have accumulated ciguatera toxin (CTX) through the marine food chain. Ciguatera fish poisoning (CFP) is a global disease, which although common, has been largely ignored in Thailand. This may be attributed to lack of confirmatory tests and seemingly nonspecific clinical presentations.

Objective: We studied CTX contamination in marine fish sold at Bangkok markets.

Methods: A surveillance of CTX in fish using the Cigua-Check assay has been conducted in Bangkok, the center of seafood marketing in Thailand. Here, there are several types of fish. Some come from domestic fishing and others are imported from Indonesia, Myanmar, or India.

Results: A random survey at three fish markets in Bangkok revealed two samples possibly contaminated with ciguatera toxin from a total 227 fish samples (of 21 fish species). This is the first report of finding CTX contamination in fish meat sold in Bangkok.

Conclusion: It is possible that these CTX-positive fish were caught from other countries. Even though the positivity (0.88%) is lower when compared with studies of other endemic areas (5.0% in Hawaii and Pacific Islands), this finding should raise awareness of the possibility of facing rare CFP intoxication from fish obtained at local Thai markets.
High glucose enhances CD39 expression in vascular endothelial cells
Authors: Kongkhum Sudawadee , Fungkrajai Mudtika , Prajan Sompoch , Bordeerat Narisa Kengtrong , Piumngam Kanyanath , Siripurkpong Pilaiwan
Published: August 2015

Background: Diabetes mellitus (DM) patients lose their ability to control normal blood glucose levels, resulting in high blood glucose levels (hyperglycemia). Hyperglycemia causes DM complications. This involves responses of vascular endothelial cells (VECs) to hyperglycemia, affecting inflammatory process and platelet activity. Ecto-enzyme CD39 is expressed on VECs, catalyzing the hydrolysis of ATP and ADP to AMP and, consequently, regulating inflammatory process and platelet activation.

Objective: We studied whether high glucose concentration has an effect on CD39 expression on VECs.

Methods: Cultured human umbilical vein endothelial cells (HUVEC) were used as a model of study. HUVEC were cultured in different glucose conditions (4, 9, 24, and 34 mM) for 24 hours. Cell viability was assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based assay and expression of CD39 was examined by using SDS-PAGE and western blot techniques.

Results: HUVEC were cultured in normal (4 and 9 mM) or high (24 and 34 mM) glucose concentrations for short term (24 hours). The results showed that high glucose (24 and 34 mM) reduced cell viability to 89.5 ± 11.3 and 86.3 ± 13.5 (mean ± SD), compared with control (4 mM), respectively. High glucose also induced increases in CD39 expression in HUVEC.

Conclusion: High glucose decreases cell viability and increases CD39 expression in HUVEC, suggesting involvement of CD39 in cell responses to high glucose.
Original article. Potential risk of dihydroergotamine causing medicationoveruse headache: preclinical evidence
Authors: Vibulyaseck Suteera , Bongsebandhu-phubhakdi Saknan , Maneesri le Grand Supang , Srikiatkhachorn Anan
Published: August 2015

Background: Overuse of abortive medication is a common factor contributing to an increase in headache frequency in patients with migraine. Whether or not chronic exposure to dihydroergotamine (DHE) can lead to this transformation remains uncertain.

Objective: To determine the effect of acute and chronic DHE exposure on development of cortical spreading depression (CSD) and trigeminal nociception.

Methods: The study comprised two experiments, namely acute and chronic exposure. In the acute experiment, a single dose of DHE (100 μg/kg) was given to male Wistar rats after successful induction of CSD. In the chronic experiment, DHE was given daily for the period of 0, 7, 14, and 28 days. CSD was induced 30 minutes after the final injection and the cortical field potential was recorded. Expression of c-Fos in caudal brainstem was used as an indicator of trigeminal nociception.

Results: Acute exposure to DHE attenuated the expression of c-Fos in the caudal brainstem without change in CSD response. By contrast, chronic exposure (14 and 28 days) to DHE increased the area under the curve of CSD waveforms. In parallel with the change in the CSD, there was significant increase of c-Fos expression within 14 days exposure to DHE and the expression remained significantly elevated for up to the 28 days examined.

Conclusion: Our study demonstrated that chronic DHE administration can increase cortical excitability and increase c-Fos expression in caudal brainstem. Our preclinical evidence suggests the possible adverse effect of chronic DHE use in causing chronification of headache.
Brief communication (Original). Rapid diagnosis of trisomy 21 by relative gene copy using real-time quantitative polymerase chain reaction
Authors: Sanguansermsri Chinnuwat , Tanpaiboon Pranoot , Charoenkwan Pimlak , Phusua Arunee
Published: August 2015

Background: Trisomy 21 or Down syndrome (DS) is the most common aneuploidy disorder. Fetal karyotypic analysis remains the criterion standard for prenatal diagnosis of DS, although the method is time consuming and requires skilled personnel. Real-time quantitative polymerase chain reaction (qPCR) can be used to determine a difference in the amount of gene copy by calculation of the difference between the cycle threshold (ΔCT) of a tested gene and a reference gene.

Objectives: To develop a rapid qPCR diagnostic method for trisomy 21.

Methods: Ten DS patients with the known karyotype of trisomy 21 were enrolled. Their parents were included as controls. D21S11 locus on chromosome 21 and SM locus on chromosome 16 from each subject were amplified by qPCR. The D21S11/SM ΔCT and 2-ΔΔCTvalues were compared between DS patients and their parents.

Results: The D21S11/SM ΔCT values of the DS patients were higher than their respective controls except for one family. The mean 2-ΔΔCTvalue between patients and mothers was 1.88 ± 0.95 (95% CI 1.20-2.56), and between fathers and mothers as controls was 1.06 ± 0.68 (95% CI 0.58-1.54).

Conclusion: The diagnostic method of trisomy 21 by using qPCR is feasible, although false negative results may occur. Using more index genes is recommended to increase the sensitivity and specificity.
Historical assessment of diphtheritic myocarditis from a hospital in northeastern Thailand
Authors: Panamonta Manat , Chaikitpinyo Arnkisa , Lumbiganon Pagakrong , Panamonta Ouyporn , Auvichayapat Narong , Wongswadiwat Yuttapong , Thepsuthammarat Kaewjai , Panthongviriyakul Aunejit , Pongchaiyakul Choowong
Published: August 2015

Background: Although there have been many descriptive studies of diphtheria from resource limited countries, descriptions of the natural history of diphtheritic myocarditis in patients from these countries are scarce.

Objective: To present the natural history of diphtheritic myocarditis from a hospital in northeastern Thailand.

Methods: The clinical features of 38 patients with diphtheria admitted to the Khon Kaen University Hospital in northeastern Thailand between 1983 and 1996 were reviewed.

Results: Of the 38 cases of diphtheria, 10 progressed to diphtheritic myocarditis (26%). Electrocardiographic findings of the 10 patients with myocarditis were myocardial and conduction abnormalities. The presence of a clinically severe (toxic) type (P < 0.001) or a swollen neck (bull neck) (P = 0.001) was a predictor of the occurrence of myocarditis. Five (50%) of the 10 patients with myocarditis had conduction abnormalities (third-degree atrioventricular block 3, left bundle branch block 1, and right bundle branch block 1). Four patients with severe symptomatic bradyarrhythmia (third-degree atrioventricular block 3, and left bundle branch block 1) received ventricular pacing, and 3 patients died after this pacing. Echocardiographic abnormalities of left ventricular dilatation and myocardial hypertrophy were found in all 5 patients with conduction abnormalities. All 6 of 7 survivors of diphtheritic myocarditis had normal 12-lead electrocardiographic results at 1-month follow-up. A patient who was the survivor of third-degree AV block had an electrocardiographic finding of flat T waves, and with a complete echocardiographic normalization of left ventricular dilatation and myocardial hypertrophy.

Conclusion: The present study confirms that increasing diphtheria immunization coverage in the population remains the most important strategy for the control of diphtheria. In resource limited countries, clinical findings, electrocardiography, and sometimes where available, echocardiography are helpful in assessing the severity of diphtheritic myocarditis, decision making of acute management, and predicting fatal outcome.
Effect of aluminum on markers of bone formation resorption in chronic hemodialysis patients
Authors: Susantitaphong Paweena , Tiranathanagul Khajohn , Katavetin Pisut , Praditpornsilpa Kearkiat , De Broe Marc E. , D’Haesec Patrick C. , Eiam-Ong Somchai
Published: August 2015

Background: T`he prevalence of aluminum (Al)-related toxicity in hemodialysis (HD) patients has declined. However, some HD patients continue to receive Al-based phosphate binders, in part because of the expense of Al-free binders.

Objective: To explore the effect of Al-based binders and their discontinuation on iron status, and markers of bone formation resorption in HD patients.

Methods: Following an initial screen of serum Al levels in 37 HD patients, a second screening was performed after discontinuation of Al-based binders in a 2-year follow-up. A desferrioxamine (DFO; 5 mg/kg) test, and assessment of iron status and bone markers were conducted in the second screening.

Results: Mean serum Al level was initially 27.8 ± 10.3 μg/L. Thirteen patients had a serum Al >30 μg/L, a level considered possibly toxic. There was a positive correlation between serum Al levels, HD duration, and cumulative dose of Al-based binder. At the second screening, the mean serum Al level decreased to 12.5 ± 7.4 μg/L. The mean serum Al level increased to 26.0 ± 14.7 μg/L post-DFO, but in none of the patients did the change in serum Al exceed the 50 μg/L threshold associated with Al-induced bone disease. The decrease in serum Al level was associated with a significant increase in intact parathyroid hormone (iPTH) whereas total alkaline phosphatase did not change.

Conclusions: We recommend that if Al-based phosphate binders are used in HD patients, serum Al level, iron, and markers of bone formation resorption be closely monitored to ensure safe use of these drugs.
RH genotypes among Malaysian blood donors
Authors: Musa Rozi Hanisa , Hassan Afifah , Ayob Yasmin , Yusoff Narazah Mohd
Published: August 2015

Background: RH genotyping studies have been conducted mainly in people of Caucasian and African descent. There is limited information regarding the molecular basis for RH genotypes in Malaysia.

Objectives: To investigate the prevalence and characteristics of RHCE genotypes among different ethnic groups in Malaysia.

Methods: A total of 1014 whole blood samples were obtained from donors from 4 different ethnic groups (360 Malays, 434 Chinese, 164 Indians, and 56 others). All samples were phenotyped for C, c, D, E, and e using standard serologic methods and genotyped using polymerase chain reaction (PCR)-based analysis.

Results: In the blood samples that we analyzed, the distribution of RH genotype antigens was significantly different among the various ethnic groups. Our findings showed that CCDee is the most common in Malaysian blood donors; 18.4% (187/1014) compared with other genotypes. The ccDEE genotype is more prevalent in the Chinese: 65.6% (82/125), and the ccee genotype is more prevalent in Indians: 47.1% (65/138). There were discrepancies between phenotypes and genotypes. There were 17 (1.7%) discrepancies in RH C/c genotyping results and of these 47% (8/17) occurred in Malays. Discrepancies in RH E/e results occurred in 3 samples (0.3%).

Conclusions: Our study provides a database for the distribution of RH genotypes of donors from the major ethnic groups in Malaysia. Methods used in this study are useful for comparing the phenotypes and genotypes. Further investigation should be conducted to study the causes of these discrepancies using other molecular based techniques.
The effect of Kodály-based music experiences on joint attention in children with autism spectrum disorders
Authors: Chiengchana Natee , Trakarnrung Somchai
Published: August 2015

Background: Kodály approach is one of widely used methods in music education and music therapy. The method is based on singing activities that could facilitate young children’s social development. Thus, the application of Kodály approach may be appropriate to enhance the joint attention behaviors of children with autism.

Objective: We examined the joint attention of individuals with autism spectrum disorders (ASD) during Kodály-based music experiences.

Methods: The A-B-C-B single-case design was employed to investigate the effectiveness of Kodály-based music experiences on joint attention behaviors of three children, ranging from 7.8 to 11.5 years of age, with autism who had manifested joint attention problems. The experimental sessions consisted of baseline condition (A), group intervention (B), and individual invention session (C). The SCERTS assessment process (SAP) was used to measure joint attention behaviors. The findings were presented using visual inspection.

Results: During baseline sessions, the three participants rarely engaged in joint attention with others. Their behaviors were stable during the first three sessions, with mean occurrence of 5, 9.3, and 4. In the course of the music intervention, they positively responded to the intervention. The frequencies of their behaviors continuously increased from the first group intervention sessions until the second group intervention sessions. Finally, the second group intervention sessions showed that most of the data points were above the mean of all previous sessions, with mean occurrence of 43.8, 34.1, and 18.8.

Conclusion: The Kodály approach was found one of useful methods in music education that can apply to music therapy to enhance joint attention behaviors of the children with autism. The children were able to learn how to engage in a group setting through singing activities, folk music, and solfège learning.
Longitudinal deficiency of upper limb: similar case presentation of two subjects with unilateral ulnar hemimelia, carpal and metacarpal deficiency, and severe oligodactyly
Authors: Afzal Muhammad , Malik Sajid
Published: August 2015

Background: Longitudinal deficiency of upper limbs with oligodactyly is a very rare congenital malformation. It manifests itself as preaxial or postaxial hypoplasia/aplasia of long bones accompanied by reduction of palm and phalanges.

Objective: To report two cases with essentially similar phenotypic presentation characterized by unilateral mesomelic shortening of limb, ulnar hypoplasia, and severe deficiency of skeletal elements of hand that were found in unrelated individuals.

Methods: Review of clinical and family history, phenotypic examination, physical and radiological investigations, and literature review.

Results: In both individuals, the right arm was short, the size of the middle arm and hand being dramatically reduced in size, and the hand comprising only two functional digits. Roentgenograms revealed hemimelia/ dysmelia of the ulna, hypoplasia of radius, dysplastic distal radial head, and several missing carpals. Only two phalangeal rays were witnessed in the hand. Radiographic measurements showed a normal contralateral arm and lower limbs, and no other associated symptoms. These phenotypes were classified as type I and type D according to the schemes proposed by Swanson et al., and Ogino and Kato, respectively. Both individuals were the product of third degree consanguineous unions (F = 0.0625).

Conclusion: Consistent phenotypic pattern of longitudinal limb anomalies evident in two independent subjects suggest a common underlying genetic etiology. There is currently no known genetic factor to allow molecular testing and risk estimation for family members. Isolated limb anomalies may provide important clues to understand pathomorphogenetic mechanisms that lead to the disruption of normal limb development.
A Study of the Relationship Between Nurses’ Professional Self-Concept and Professional Ethics in Hospitals Affiliated to Jahrom University of Medical Sciences, Iran
Authors: Parandavar Nehleh , Rahmanian Afifeh , Badiyepeymaie Jahromi Zohreh
Published: August 2015

BACKGROUND: Commitment to ethics usually results in nurses’ better professional performance and advancement. Professional self-concept of nurses refers to their information and beliefs about their roles, values, and behaviors. The objective of this study is to analyze the relationship between nurses’ professional self-concept and professional ethics in hospitals affiliated to Jahrom University of Medical Sciences.

METHODS: This cross sectional-analytical study was conducted in 2014. The 270 participants were practicing nurses and head-nurses at the teaching hospitals of Peimanieh and Motahari in Jahrom University of Medical Science. Sampling was based on sencus method. Data was collected using Cowin’s Nurses’ self-concept questionnaire (NSCQ) and the researcher-made questionnaire of professional ethics.

RESULTS: The average of the sample’s professional self-concept score was 6.48±0.03 out of 8. The average of the sample’s commitment to professional ethics score was 4.08±0.08 out of 5. Based on Pearson’s correlation test, there is a significant relationship between professional ethics and professional self-concept (P=0.01, r=0.16).

CONCLUSION: In view of the correlation between professional self-concept and professional ethics, it is recommended that nurses’ self-concept, which can boost their commitment to ethics, be given more consideration.
Beauty in the eye of the beholder? Attractiveness in a virtual world
Authors: Guan Shu-Sha Angie , Subrahmanyam Kaveri , Linares Kevin , Cheng Roy
Published: August 2015

The current study examines whether traditional offline gender biases regarding physical attractiveness and status transfer to Second Life (SL), a virtual world where residents can easily manipulate the appearance and status of avatars (i.e., graphical representations of the self). Participants (N = 312, 60.58% female, Mage = 29.77, SD = 10.53) reported on demographics, SL usage, and rated the attractiveness of female and male avatars manipulated along physical attractiveness (high vs. low) and status (high vs. low). Mixed measures ANCOVAs were modeled separately for the female avatars and male avatars with within-subjects factors (avatar characteristics: physical attractiveness and status) and between-subject factors (participant characteristics: sex, and intensity of SL usage), while controlling for participant age. Consistent with offline norms, female avatars high in physical attractiveness were rated as more attractive, regardless of status. Participants rated male avatars high in physical attractiveness as more attractive if they were high in status compared to those low in status. We also found opposite-sex preferences and moderations by participant age and SL usage on avatar attractiveness ratings. The results suggest the continuity of offline gender norms and effect of in-world experience on perceptions of avatar appearance.
Parlamentary elections in the District of Berane in 1920 and 1923
Authors: Virijević Vladan , Deletić Zdravko
Published: August 2015

The lack of more significant political tradition due to the fact that Berane and the area of Donji Vasojevici were liberated from the Ottoman authorities in 1912, which lasted for many centuries, as well as the fact that only three years later a two-year period of AustroHungarian occupation took place, mainly predetermined the atmosphere of political life in the first years after Yugoslav unification in 1918. The underdevelopment of the party organizations and grouping based on denominational, tribal and even on belonging to some brotherhood, were characteristic for the beginnings of political subjects in Berane district. These were party organizations whose headquarters were settled in Belgrade, Sarajevo and Skopje and whose leaders were as in the case of other districts in the province, often resorted to emphasizing their candidature in them. There were also the individuals on the local political scene, mostly teachers or the civil servants, who were representatives of the party's ideas and programme in their surrounding and therefore also the incumbent candidates of their lists. During parliamentary elections in Berane in 1920 and 1923 the main political rivalry was between members of the radical and democratic party while it was characterised by fragmentation of the party namely emphasizing 'dissident' candidate list as the consequences of political ambitions of the individuals. Regarding the results of elections in 1920 and 1923, generally speaking, it can be said that during the first half of the 20s of the last century Democratic party dominated in Berane district. Berane got 'its' Member of the Parliament - 'a democrat' Pavle Cubrovic, PhD while in 1923 it no longer had its representative in the parliament. Namely, despite Cubrovic's triumph in Berane district, since his candidate list did not get enough votes in the first round of elections, the position of the Member of the Parliament 'escaped' him.
On roots and nature of the normative regulation of the criminal offence of theft in the tradition of Tsar Dusan legislation
Authors: Vujović Ognjen
Published: August 2015

In this paper the comparison between several norms of the Crimi­nal Offence of Theft in the tradition of Tsar Dusan legislation and certain provisions of the Roman law was performed. On this occa­sion, the issue of robbery, theft of fruits and complicity to theft were treated in particular.
Três percepções sobre gestação e puerpério
Authors: Fernandes César Eduardo
Published: August 2015

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Da incredibile a credibile: strategie nazionali di promozione turistica in India
Authors: Maria Angelillo
Published: August 2015

Tourism is at the top of Indian current Prime Minister Narendra Modi’s agenda: according to Bharatiya Janata Party’s manifesto, tourism can play a key role in India’s socio-economic progress. The country had last formulated a tourism policy in 2002, when the multimillion dollar nation branding campaign which received the name “Incredible India” was launched on an international level. The primary objective of this branding exercise was to create a distinctive identity for the country. Through the analysis of the campaign’s themes, the paper discusses its connection with orientalist imagery and colonial meanings as well as its effectiveness in promoting the Indian tourist industry. The article dwells on how the government’s campaign constructs the Indian difference for an imagined global audience. Questions of cultural othering, the role of colonialism, and contemporary neocolonial structures in relation to tourism promotion for India are discussed. In this paper it is argued that, if the government is serious about branding India with Narendra Modi’s much hyped 5 Ts, with tourism as a leading contributor, a new paradigm different from the “Incredible India” one may be necessary.
It's all starting to come together
Authors: Becker Lindsay A , Gitler Aaron D
Published: August 2015

Chemical, genetic and cell biology tools have been used to probe which RNA-protein granules behave like liquids and which behave like solids.
Overview of Probing Protein‐Ligand Interactions Using NMR
Authors: Aguirre Clémentine , Cala Olivier , Krimm Isabelle
Published: August 2015

Nuclear magnetic resonance (NMR) is a powerful technique for the study and characterization of protein‐ligand interactions. In this unit we review both experiments where the NMR spectrum of the protein is observed (protein‐observed NMR experiments) and those where the NMR spectra of the ligand is observed (ligand‐observed NMR experiments) for the identification of binding partners, the measurement of protein‐ligand affinity, the design of molecules that are active against biological targets such as proteins, and the assessment of the binding modes of the ligands. Ligand‐observed methods discussed in this unit are Nuclear Overhauser Effect (NOE)—based approaches, with well‐known experiments such as the Saturation Transfer Difference, Water‐Ligand Observed via Gradient Spectroscopy (WaterLOGSY), and transferred—Nuclear Overhauser Effect Spectroscopy (tr‐NOESY) experiments, and also the INPHARMA experiment. Regarding protein‐observed experiments, this unit focuses on the use of chemical shift perturbations observed in protein‐NMR spectra upon ligand binding. Also discussed is how these chemical shift perturbations can be used for the analysis of protein‐ligand complexes, including fast structure determination when combined with docking. © 2015 by John Wiley & Sons, Inc.
Using Single Lectins to Enrich Glycoproteins in Conditioned Media
Authors: Sethi Manveen K. , Fanayan Susan
Published: August 2015

Lectins are sugar‐binding proteins that can recognize and bind to carbohydrates conjugated to proteins and lipids. Coupled with mass spectrometry technologies, lectin affinity chromatography is becoming a popular approach for identification and quantification of glycoproteins in complex samples such as blood, tumor tissues, and cell lines. Given the commercial availability of a large number of lectins that recognize diverse sugar structures, it is now possible to isolate and study glycoproteins for biological and medical research. This unit provides a general guide to single‐lectin‐based enrichment of glycoproteins from serum‐free conditioned media. Due to the unique carbohydrate specificity of most lectins and the complexity of the samples, optimization steps may be required to evaluate different elution buffers and methods as well as binding conditions, for each lectin, for optimal recovery of bound glycoproteins. © 2015 by John Wiley & Sons, Inc.
Applications of Lipid Nanodiscs for the Study of Membrane Proteins by Surface Plasmon Resonance
Authors: Trahey Meg , Li Mavis Jiarong , Kwon Hyewon , Woodahl Erica L. , McClary Wynton D. , Atkins William M.
Published: August 2015

Methods for the initial steps of surface plasmon resonance analysis of membrane proteins incorporated in lipid nanodiscs are described. Several types of Biacore sensor chips are available and require distinct strategies to immobilize proteonanodiscs on the chip surface. The procedures for immobilization on three of these chips (NTA, antibody coupled CM5, and L1) are described in this unit and results are demonstrated for a model system with cytochrome P4503A4 (CYP3A4) in nanodiscs binding to a polyclonal anti‐CYP3A4 antibody. Advantages and disadvantages of each chip type are considered. © 2015 by John Wiley & Sons, Inc.
Isolation of Mouse Neutrophils
Authors: Swamydas Muthulekha , Luo Yi , Dorf Martin E. , Lionakis Michail S.
Published: August 2015

Neutrophils represent the first line of defense against bacterial and fungal pathogens. Indeed, patients with inherited and acquired qualitative and quantitative neutrophil defects are at high risk for developing bacterial and fungal infections and suffering adverse outcomes from these infections. Therefore, research aiming at defining the molecular factors that modulate neutrophil effector function under homeostatic conditions and during infection is essential for devising strategies to augment neutrophil function and improve the outcome of infected individuals. This unit describes a reproducible density gradient centrifugation‐based protocol that can be applied in any laboratory to harvest large numbers of highly enriched and highly viable neutrophils from the bone marrow of mice both at the steady state and following infection with Candida albicans as described in
UNIT
. In another protocol, we also present a method that combines gentle enzymatic tissue digestion with a positive immunomagnetic selection technique or Fluorescence‐activated cell sorting (FACS) to harvest highly pure and highly viable preparations of neutrophils directly from mouse tissues such as the kidney, the liver or the spleen. Finally, methods for isolating neutrophils from mouse peritoneal fluid and peripheral blood are included. Mouse neutrophils isolated by these protocols can be used for examining several aspects of cellular function ex vivo including pathogen binding, phagocytosis and killing, neutrophil chemotaxis, oxidative burst, degranulation and cytokine production, and for performing neutrophil adoptive transfer experiments. © 2015 by John Wiley & Sons, Inc.
Magnetospheric ULF waves with increasing amplitude related to solar wind dynamic pressure changes: The Time History of Events and Macroscale Interactions during Substorms (THEMIS) observations
Authors: Shen X. C. , Zong Q.‐G. , Shi Q. Q. , Tian A. M. , Sun W. J. , Wang Y. F. , Zhou X. Z. , Fu S. Y. , Hartinger M. D. , Angelopoulos V.
Published: August 2015

Ultralow frequency (ULF) waves play an important role in transferring energy by buffeting the magnetosphere with solar wind pressure impulses. The amplitudes of magnetospheric ULF waves, which are induced by solar wind dynamic pressure enhancements or shocks, are thought to damp in one half a wave cycle or an entire wave cycle. We report in situ observations of solar wind dynamic pressure impulse‐induced magnetospheric ULF waves with increasing amplitudes. We found six ULF wave events induced by solar wind dynamic pressure enhancements with slow but clear wave amplitude increase. During three or four wave cycles, the amplitudes of ion velocities and electric field of these waves increased continuously by 1.3–4.4 times. Two significant events were selected to further study the characteristics of these ULF waves. We found that the wave amplitude growth is mainly contributed by the toroidal mode wave. Three possible mechanisms of causing the wave amplitude increase are discussed. First, solar wind dynamic pressure perturbations, which are observed in a duration of 20–30 min, might transfer energy to the magnetospheric ULF waves continually. Second, the wave amplitude increase in the radial electric field may be caused by superposition of two wave modes, a standing wave excited by the solar wind dynamic impulse and a propagating compressional wave directly induced by solar wind oscillations. When superposed, the two wave modes fit observations as does a calculation that superposes electric fields from two wave sources. Third, the normal of the solar wind discontinuity is at an angle to the Sun‐Earth line. Thus, the discontinuity will affect the dayside magnetopause continuously for a long time.
Retrieving 65 years of volcano summit deformation from multitemporal structure from motion: The case of Piton de la Fournaise (La Réunion Island)
Authors: Derrien Allan , Villeneuve Nicolas , Peltier Aline , Beauducel François
Published: August 2015

The structure from motion photogrammetry technique enables use of historical airborne photography to achieve high‐resolution 3‐D terrain models. We apply this method on Piton de la Fournaise volcano (La Réunion), which allows a unique opportunity to retrieve high‐resolution (1.5–0.11 m) digital elevation models and precise deformation maps of the volcano since 1950. Our results provide evidence that the summit volume increased throughout the study period, at a stable rate of 2.2 Mm3/yr between 1950 and 2015, with an acceleration up to 8.0 Mm3/yr prior to the major 2007 eruption that was accompanied by summit caldera collapse. At the same time, summit deformation was asymmetric, with 9.2 ± 2.5 m of eastward seaward displacement and 1.3 ± 2.5 m to the west during 1950–2015. Our results reveal a temporal evolution in the volcano magma influx rate and deformation. Tracking these fluxes and the long‐lived preferential eastern motion is crucial to mitigate risks associated to flank destabilization.
Water exchange between the continental shelf and the cavity beneath Nioghalvfjerdsbræ (79 North Glacier)
Authors: Wilson N. J. , Straneo F.
Published: August 2015

The mass loss at Nioghalvfjerdsbræ is primarily due to rapid submarine melting. Ocean data obtained from beneath the Nioghalvfjerdsbræ ice tongue show that melting is driven by the presence of warm (1°C) Atlantic Intermediate Water (AIW). A sill prevents AIW from entering the cavity from Dijmphna Sund, requiring that it flow into the cavity via bathymetric channels to the south at a pinned ice front. Comparison of water properties from the cavity, Dijmphna Sund, and the continental shelf support this conclusion. Overturning circulation rates inferred from observed melt rates and cavity stratification suggest an exchange flow between the cavity and the continental shelf of 38mSv, sufficient to flush cavity waters in under 1 year. These results place upper bounds on the timescales of external variability that can be transmitted to the glacier via the ice tongue cavity.
Generation of large postinjection‐induced seismic events by backflow from dead‐end faults and fractures
Authors: McClure Mark W.
Published: August 2015

Induced seismicity is common during hydraulic stimulation in fractured crystalline rock. Fluid injection pressurizes preexisting fractures, triggering slip and seismicity. Often, the largest induced events occur after the end of injection, which complicates efforts to manage seismic risk. In this study, a three‐dimensional discrete fracture network simulator that couples fluid flow with earthquake simulation was used to investigate a novel hypothesis for why large postinjection seismic events occur. Fractures that form dead‐end pathways differentially pressurize during injection. After injection is stopped, fluid backflows through the well from the dead‐end fractures into larger fractures, inducing additional seismicity and potentially causing events larger than occurred during injection. Our simulations indicate that flowing fluid back to the surface immediately after injection could mitigate this effect and reduce postinjection seismicity.
Seasonal drought predictability and forecast skill over China
Authors: Ma Feng , Yuan Xing , Ye Aizhong
Published: August 2015

Under a changing environment, seasonal droughts have been exacerbated with devastating impacts. However, the understanding of drought mechanism and predictability is limited. Based on the hindcasts from multiple climate models, the predictability and forecast skill for drought over China are investigated. The 3 month standardized precipitation index is used as the drought index, and the predictability is quantified by using a perfect model assumption. Ensemble hindcasts from multiple climate models are assessed individually, and the grand multimodel ensemble is also evaluated. Drought forecast skill for model ensemble mean is higher than individual ensemble members, and North American Multimodel Ensemble grand ensemble performs the best. Predictability is higher than forecast skill, indicating the room for improving drought forecast. Drought predictability and forecast skill are positively correlated in general, but they vary depending on seasons, regions, and forecast leads. Higher drought predictability and forecast skill are found over regimes where ENSO has significant impact. For the ENSO‐affected regimes, both drought predictability and forecast skill in ENSO years are higher than that in neutral years. This study suggests that predictability not only provides a measure for selecting climate models for ensemble drought forecast in ENSO‐affected regimes but also serves as an indicator for forecast skill especially when in situ and/or remote sensing measurements for the hindcast verifications are considered unreliable.
The influence of marine microbial activities on aerosol production: A laboratory mesocosm study
Authors: Alpert Peter A. , Kilthau Wendy P. , Bothe Dylan W. , Radway JoAnn C. , Aller Josephine Y. , Knopf Daniel A.
Published: August 2015

The oceans cover most of the Earth's surface, contain nearly half the total global primary biomass productivity, and are a major source of atmospheric aerosol particles. Here we experimentally investigate links between biological activity in seawater and sea spray aerosol (SSA) flux, a relationship of potential significance for organic aerosol loading and cloud formation over the oceans and thus for climate globally. Bubbles were generated in laboratory mesocosm experiments either by recirculating impinging water jets or glass frits. Experiments were conducted with Atlantic Ocean seawater collected off the eastern end of Long Island, NY, and with artificial seawater containing cultures of bacteria and phytoplankton Thalassiosira pseudonana, Emiliania huxleyi, and Nannochloris atomus. Changes in SSA size distributions occurred during all phases of bacterial and phytoplankton growth, as characterized by cell concentrations, dissolved organic carbon, total particulate carbon, and transparent exopolymer particles (gel‐forming polysaccharides representing a major component of biogenic exudate material). Over a 2 week growth period, SSA particle concentrations increased by a factor of less than 2 when only bacteria were present and by a factor of about 3 when bacteria and phytoplankton were present. Production of jet‐generated SSA particles of diameter less than 200 nm increased with time, while production of all particle diameters increased with time when frits were used. The implications of a marine biological activity dependent SSA flux are discussed.
Isolation of Undifferentiated Female Germline Cells from Adult Drosophila Ovaries
Authors: Lim Robyn Su May , Osato Motomi , Kai Toshie
Published: August 2015

This unit describes a method for isolating undifferentiated, stem cell–like germline cells from adult Drosophila ovaries. Here, we demonstrate that this population of cells can be effectively purified from hand‐dissected ovaries in considerably large quantities. Tumor ovaries with expanded populations of undifferentiated germline cells are first removed from fly abdomens and dissociated into a cell suspension with the aid of protease treatment. The target cells, which express Vasa‐green fluorescent protein (GFP) fusion protein under the control of the germline‐specific vasa promoter, are specifically selected from the suspension via fluorescence‐activated cell sorting (FACS). These protocols can be adapted to isolate other cell types from fly ovaries, such as somatic follicle cells or escort cells, by driving GFP expression in the respective target cells. © 2015 by John Wiley & Sons, Inc.
Regional Cell Specific RNA Expression Profiling of FACS Isolated Drosophila Intestinal Cell Populations
Authors: Dutta Devanjali , Buchon Nicolas , Xiang Jinyi , Edgar Bruce A.
Published: August 2015

The adult Drosophila midgut is built of five distinct cell types, including stem cells, enteroblasts, enterocytes, enteroendocrine cells, and visceral muscles, and is divided into five major regions (R1 to R5), which are morphologically and functionally distinct from each other. This unit describes a protocol for the isolation of Drosophila intestinal cell populations for the purpose of cell type–specific transcriptome profiling from the five different regions. A method to select a cell type of interest labeled with green or yellow fluorescent protein (GFP, YFP) by making use of the GAL4‐UAS bipartite system and fluorescent‐activated cell sorting (FACS) is presented. Total RNA is isolated from the sorted cells of each region, and linear RNA amplification is used to obtain sufficient amounts of high‐quality RNA for analysis by microarray, RT‐PCR, or RNA sequencing. This method will be useful for quantitative transcriptome comparison across intestinal cell types in the different regions under normal and various experimental conditions. © 2015 by John Wiley & Sons, Inc.
TALEN‐ and CRISPR/Cas9‐Mediated Gene Editing in Human Pluripotent Stem Cells Using Lipid‐Based Transfection
Authors: Hendriks William T. , Jiang Xin , Daheron Laurence , Cowan Chad A.
Published: August 2015

Using custom‐engineered nuclease‐mediated genome editing, such as Transcription Activator–Like Effector Nucleases (TALENs) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) RNA‐guided Cas9 nucleases, human pluripotent stem cell (hPSC) lines with knockout or mutant alleles can be generated and differentiated into various cell types. This strategy of genome engineering in hPSCs will prove invaluable for studying human biology and disease. Here, we provide a detailed protocol for design and construction of TALEN and CRISPR vectors, testing of their nuclease activity, and delivery of TALEN or CRISPR vectors into hPSCs. In addition, we describe the use of single‐stranded oligodeoxynucleotides (ssODNs) to introduce or repair point mutations. Next, we describe the identification of edited hPSC clones without antibiotic selection, including their clonal selection, genotyping, and expansion for downstream applications. © 2015 by John Wiley & Sons, Inc.
Hepatocyte‐Based Metabolism, Drug–Drug Interaction, and Toxicity Assays in Drug Discovery and Drug Development to Minimize Attrition
Authors: Li Albert P.
Published: August 2015

Primary human hepatocytes are generally considered the “gold standard” forin vitroevaluation of human drug metabolism, drug‐drug interactions (DDIs), and hepatotoxicity. Successful cryopreservation to retain high viability and plateability (ability to be cultured) allows human hepatocytes to be used routinely for experimentation. Assays developed with human hepatocytes include metabolic stability, metabolite profiling, P450 inhibition, P450 induction, andin vitrohepatotoxicity. These assays are routinely used in drug development to minimize metabolic and safety liability of new chemical entities (NCEs).