Molecular Breeding for Ascochyta Blight Resistance in Lentil: Current Progress and Future Directions.

Abstract: Lentil (Lens culinaris Medik.) is a diploid (2n = 2x = 14), self-pollinating, cool-season, grain legume that is cultivated worldwide and is highly valuable due to its high protein content. However, lentil production is constrained by many factors including biotic stresses, majority of which are fungal diseases such as ascochyta blight (AB), fusarium wilt, rust, stemphylium blight, anthracnose, and botrytis gray mold. Among various diseases, AB is a major -problem in many lentil-producing countries and can significantly reduce crop production. Breeding for AB resistance has been a priority for breeding programs across the globe and consequently, a number of resistance sources have been identified and extensively exploited. In order to increase the efficiency of combining genes from different genetic backgrounds, molecular genetic tools can be integrated with conventional breeding methods. A range of genetic linkage maps have been generated based on DNA-based markers, and quantitative trait loci (QTLs) for AB resistance have been identified. Molecular markers linked to these QTLs may potentially be used for efficient pyramiding of the AB disease resistance genes. Significant genomic resources have been established to identify and characterize resistance genes, including an integrated genetic map, expressed sequence tag libraries, gene based markers, and draft genome sequences. These resources are already being utilized for lentil crop improvement, to more effectively select for disease resistance, as a case study of the Australian breeding program will show. The combination of genomic resources, effective molecular genetic tools and high resolution phenotyping tools will improve the efficiency of selection for ascochyta blight resistance and accelerate varietal development of global lentil breeding programs.

Rodda|Matthew S|MS|
Davidson|Jennifer|J|
Javid|Muhammad|M|
Sudheesh|Shimna|S|
Blake|Sara|S|
Forster|John W|JW|
Kaur|Sukhjiwan|S|

legume
mapping
molecular markers
fungal disease resistance
pulse


Production of the Fusarium Mycotoxin Moniliformin by Penicillium melanoconidium.

Abstract: Moniliformin is a mycotoxin produced by several cereal associated Fusaria. Here, we show for the first time that moniliformin can be produced by the cereal fungus, Penicillium melanoconidium (4 out of 4 strains), but not in the related species in the Viridicata series. Moniliformin was detected in 10 out of 11 media: two agars and several cereal and bean types. Moniliformin was identified by a novel mixed-mode anionic exchange reversed phase chromatographic method which was coupled to both tandem mass spectrometry (MS) and high resolution MS. Mixed-mode chromatography showed superior peak shape compared to that of HILIC and less matrix interference compared to that of reversed phase chromatography, but during a large series of analyses, the column was fouled by matrix interferences. Wheat and beans were artificially infected by P. melanoconidium containing up to 64 and 11 mg/kg moniliformin, respectively, while penicillic acid, roquefortine C, and penitrem A levels in wheat were up to 1095, 38, and 119 mg/kg, respectively.

Hallas-Møller|Magnus|M|
Nielsen|Kristian Fog|KF|
Frisvad|Jens Christian|JC|

D002851:Chromatography, High Pressure Liquid

moniliformin
Penicillium melanoconidium
mixed-mode chromatography
penicillic acid
roquefortine C
penitrem A
LC-MS/MS
wheat grain
white beans


Prediction of Early Childhood Caries via Spatial-Temporal Variations of Oral Microbiota.

Abstract: Microbiota-based prediction of chronic infections is promising yet not well established. Early childhood caries (ECC) is the most common infection in children. Here we simultaneously tracked microbiota development at plaque and saliva in 50 4-year-old preschoolers for 2 years; children either stayed healthy, transitioned into cariogenesis, or experienced caries exacerbation. Caries onset delayed microbiota development, which is otherwise correlated with aging in healthy children. Both plaque and saliva microbiota are more correlated with changes in ECC severity (dmfs) during onset than progression. By distinguishing between aging- and disease-associated taxa and exploiting the distinct microbiota dynamics between onset and progression, we developed a model, Microbial Indicators of Caries, to diagnose ECC from healthy samples with 70% accuracy and predict, with 81% accuracy, future ECC onsets for samples clinically perceived as healthy. Thus, caries onset in apparently healthy teeth can be predicted using microbiota, when appropriately de-trended for age.

Teng|Fei|F|
Yang|Fang|F|
Huang|Shi|S|
Bo|Cunpei|C|
Xu|Zhenjiang Zech|ZZ|
Amir|Amnon|A|
Knight|Rob|R|
Ling|Junqi|J|
Xu|Jian|J|

D002675:Child, Preschool
D004269:DNA, Bacterial
D004275:DNA, Ribosomal
D003731:Dental Caries
D003773:Dental Plaque
D006801:Humans
D064307:Microbiota
D015233:Models, Statistical
D008969:Molecular Sequence Data
D011379:Prognosis
D012336:RNA, Ribosomal, 16S
D012463:Saliva
D017422:Sequence Analysis, DNA

microbiota
plaque
saliva
microbial indicators of caries
early-childhood caries


Longitudinal Analysis of Music Education on Executive Functions in Primary School Children.

Abstract: Background: Research on the effects of music education on cognitive abilities has generated increasing interest across the scientific community. Nonetheless, longitudinal studies investigating the effects of structured music education on cognitive sub-functions are still rare. Prime candidates for investigating a relationship between academic achievement and music education appear to be executive functions such as planning, working memory, and inhibition. Methods: One hundred and forty-seven primary school children, Mage = 6.4 years, SD = 0.65 were followed for 2.5 years. Participants were randomized into four groups: two music intervention groups, one active visual arts group, and a no arts control group. Neuropsychological tests assessed verbal intelligence and executive functions. Additionally, a national pupil monitor provided data on academic performance. Results: Children in the visual arts group perform better on visuospatial memory tasks as compared to the three other conditions. However, the test scores on inhibition, planning and verbal intelligence increased significantly in the two music groups over time as compared to the visual art and no arts controls. Mediation analysis with executive functions and verbal IQ as mediator for academic performance have shown a possible far transfer effect from executive sub-function to academic performance scores. Discussion: The present results indicate a positive influence of long-term music education on cognitive abilities such as inhibition and planning. Of note, following a two-and-a-half year long visual arts program significantly improves scores on a visuospatial memory task. All results combined, this study supports a far transfer effect from music education to academic achievement mediated by executive sub-functions.

Jaschke|Artur C|AC|
Honing|Henkjan|H|
Scherder|Erik J A|EJA|

longitudinal analysis
music
executive functions
intelligence
far transfer


The effect of task order predictability in audio-visual dual task performance: Just a central capacity limitation?

Abstract: In classic Psychological-Refractory-Period (PRP) dual-task paradigms, decreasing stimulus onset asynchronies (SOA) between the two tasks typically lead to increasing reaction times (RT) to the second task and, when task order is non-predictable, to prolonged RTs to the first task. Traditionally, both RT effects have been advocated to originate exclusively from the dynamics of a central bottleneck. By focusing on two specific electroencephalographic brain responses directly linkable to perceptual or motor processing stages, respectively, the present study aimed to provide a more detailed picture as to the origin(s) of these behavioral PRP effects. In particular, we employed 2-alternative forced-choice (2AFC) tasks requiring participants to identify the pitch of a tone (high versus low) in the auditory, and the orientation of a target object (vertical versus horizontal) in the visual, task, with task order being either predictable or non-predictable. Our findings show that task order predictability (TOP) and inter-task SOA interactively determine the speed of (visual) perceptual processes (as indexed by the PCN timing) for both the first and the second task. By contrast, motor response execution times (as indexed by the LRP timing) are influenced independently by TOP for the first, and SOA for the second, task. Overall, this set of findings complements classical as well as advanced versions of the central bottleneck model by providing electrophysiological evidence for modulations of both perceptual and motor processing dynamics that, in summation with central capacity limitations, give rise to the behavioral PRP outcome.

Töllner|Thomas|T|
Strobach|Tilo|T|
Schubert|Torsten|T|
Müller|Hermann J|HJ|

central bottleneck
PCN
LRP
attention
decision making
executive control


Safety and efficacy of enhanced recovery after surgery in elderly patients after therapeutic endoscopic retrograde cholangiopancreatography.

Abstract:
Introduction
The number of elderly patients with biliary and pancreatic diseases has increased significantly. The characteristics of biliary and pancreatic diseases in the elderly increase the risk of treatment.

Aim
To study the safety and efficacy of therapeutic endoscopic retrograde cholangiopancreatography (ERCP) in elderly patients with biliary and pancreatic diseases with the concept of enhanced recovery after surgery (ERAS).

Material and methods
Patients receiving ERCP under ERAS were grouped into an elderly group (group A, n = 58, aged 75 years or above) and a young and middle-aged group (group B, n = 202, aged less than 60 years). The clinical parameters before, during and after the operation of the two groups were compared.

Results
Before the operation, the incidences of cholangiocarcinoma and complications, nutritional screening score ≥ 3, ASA degree III and Child-Pugh grade A in group A were significantly higher than those in group B (p < 0.05), while the incidences of nausea and vomiting, abdominal pain, nutritional screening < 3 and ASA grade I in group A were significantly lower than those in group B (p < 0.05). Intraoperatively, the incidence of juxta-ampullary duodenal diverticulum (JAD) in internal or bottom papilla in the elder patients with difficult selective biliary cannulation (DSBC) was significantly higher than that in the young and middle-aged group (p < 0.05). In addition, the continuous ECG monitoring duration and the first exhaust time in group A were significantly longer than those in group B (p < 0.05).

Conclusions
Endoscopic retrograde cholangiopancreatography under ERAS in elderly patients is as safe and effective as in young patients.

Niu|Fang|F| Liu|Yu-Di|YD| Chen|Rong-Xia|RX| Niu|Yu-Jun|YJ|


Reliability and validity of the self-reported frailty screening questionnaire in older adults.

Abstract:
Background
Frailty is one of the most important risk factors for adverse outcomes in older adults. Despite a high prevalence, there is still a lack of frailty screening instruments specific to Chinese older adults. We developed a simple frailty screening questionnaire (FSQ) that could predict long-term mortality. We aimed to explore the reliability and construct validity of this new measurement tool.

Methods
A total of 205 individuals aged 65 years or older were recruited in this study. The FSQ and frailty phenotype were assessed. The FSQ included self-reported slowness, weakness, weight loss, inactivity, and exhaustion. A subgroup of 109 participants completed the FSQ a second time 2 weeks later for test-retest reliability. Frailty phenotype included slowness, exhaustion, weight loss, weakness, and inactivity.

Results
The intraclass correlation coefficient for the FSQ, slowness, weakness, weight loss, inactivity and exhaustion were 0.937, 0.938, 0.934, 0.921, 0.826, and 0.832, respectively. Using a cut-off of 3, the sensitivity, specificity, and area under the curve of the receiver operating characteristic were 52.6%, 93.5%, and 0.883 (p < 0.001), respectively. The kappa coefficient between the FSQ and frailty phenotype was 0.431 (p < 0.001). FSQ score was negatively correlated with walking speed and grip strength, and positively correlated with age. Frailty defined by the FSQ was associated with older age, chronic diseases, and worse physical function.

Conclusions
The FSQ is a potentially useful, reliable, and valid instrument in screening frailty in older adults, and can be recommended to identify frailty in clinical settings.


Characterization of Amikacin Drug Exposure and Nephrotoxicity in an Animal Model.

Abstract: Despite excellent in vitro activity, aminoglycosides are used conservatively to treat multidrug-resistant bacterial infections due to their associated nephrotoxicity. Aminoglycosides are known to accumulate in the kidneys, but the quantitative relationship between drug exposures and nephrotoxicity is not well established. To bridge the knowledge gap, the objective of this study was to develop an animal model with clinically relevant conditions to mimic human disease progression. Single-dose pharmacokinetics were studied in Sprague-Dawley rats dosed either with 100 or 500 mg/kg of body weight of amikacin subcutaneously. Serial blood samples were collected, and serum amikacin concentrations were measured using liquid chromatography tandem mass spectrometry. Rats were also dosed with amikacin once daily for up to 10 days; blood samples were taken at baseline and daily to detect nephrotoxicity (defined as doubling of serum creatinine from baseline). Kidneys from both studies were harvested from selected rats, and amikacin concentrations in renal tissues were measured. A dose-dependent increase in systemic area under the curve (AUC) was observed, which ranged from approximately 1/3 (AUC of 53 mg·h/liter) to 3 times (AUC of 650 mg·h/liter) the expected exposure resulting from standard dosing in humans. Nephrotoxicity was significantly higher in rats given 500 mg/kg (100% versus 30%, P = 0.003). Kaplan-Meier analysis also showed a significant difference in nephrotoxicity onset between the two groups (P = 0.001). Finally, analysis of the renal tissues showed that the accumulation of amikacin could be associated with nephrotoxicity. These results are consistent with clinical observations, which support using this model in the future to investigate an intervention(s) that can be used clinically to alleviate nephrotoxicity.

Chan|Katrina|K|
Ledesma|Kimberly R|KR|
Wang|Weiqun|W|
Tam|Vincent H|VH|0000-0001-7071-5376

aminoglycosides
pharmacokinetics
liquid chromatography
acute kidney injury
high performance liquid chromatography


Primary tuberculosis in a malnourished adolescent.

Abstract: Although the overall prevalence of tuberculosis has decreased in the United States, with the increasing prevalence of tuberculosis globally, higher rates of tuberculosis in some states and localities have been reported, with some component probably related to immigrant populations. We report a case of primary pulmonary tuberculosis in a malnourished adolescent.

Zawaideh|Mazen|M| Chao|Cherng|C| Poole|Patricia|P| Naheedy|John|J|


miR-4513 promotes breast cancer progression through targeting TRIM3.

Abstract: The biological function of microRNA-4513 (miR-4513) in human cancers is emerging. However, it remains unknown whether miR-4513 has a role in breast cancer (BC). In this study, we analyzed the expression of miR-4513 and tripartite motif containing 3 (TRIM3) in BC cell lines. The biological roles of miR-4513 and TRIM3 in BC were analyzed by cell counting kit-8 assay, colony formation assay, wound healing assay, and transwell invasion assay. The effects of miR-4513 or TRIM3 expression on the overall survival of BC patients were analyzed at Kaplan-Meier plotter website. We found miR-4513 expression was upregulated, whereas TRIM3 expression was downregulated in BC cell lines. Importantly, high miR-4513 or low TRIM3 expression was revealed as predictors for poorer overall survival of BC patients. Luciferase activity assay and western blot assay revealed TRIM3 was a direct target of miR-4513. Moreover, we showed miR-4513 was able to regulate BC cell proliferation, colony formation, cell migration, and cell invasion through regulating TRIM3. Taken together, our results suggested miR-4513 functions as an oncogene in the progression of BC and, therefore, miR-4513 may be validated as a potential therapeutic target in the future.

Li|Yongzhen|Y| Zhu|Huifang|H| Wang|Jianqiang|J| Qian|Xinlai|X| Li|Na|N|


What makes Xanthomonas albilineans unique amongst xanthomonads?

Abstract: Xanthomonas albilineans causes leaf scald, a lethal disease of sugarcane. Compared to other species of Xanthomonas, X. albilineans exhibits distinctive pathogenic mechanisms, ecology and taxonomy. Its genome, which has experienced significant erosion, has unique genomic features. It lacks two loci required for pathogenicity in other plant pathogenic species of Xanthomonas: the xanthan gum biosynthesis and the Hrp-T3SS (hypersensitive response and pathogenicity-type three secretion system) gene clusters. Instead, X. albilineans harbors in its genome an SPI-1 (Salmonella pathogenicity island-1) T3SS gene cluster usually found in animal pathogens. X. albilineans produces a potent DNA gyrase inhibitor called albicidin, which blocks chloroplast differentiation, resulting in the characteristic white foliar stripe symptoms. The antibacterial activity of albicidin also confers on X. albilineans a competitive advantage against rival bacteria during sugarcane colonization. Recent chemical studies have uncovered the unique structure of albicidin and allowed us to partially elucidate its fascinating biosynthesis apparatus, which involves an enigmatic hybrid PKS/NRPS (polyketide synthase/non-ribosomal peptide synthetase) machinery.

Pieretti|Isabelle|I|
Pesic|Alexander|A|
Petras|Daniel|D|
Royer|Monique|M|
Süssmuth|Roderich D|RD|
Cociancich|Stéphane|S|

Xanthomonas albilineans
genomic features
albicidin
NRPS and PKS genes
leaf scald disease of sugarcane


Exploiting a low tax system: non-tax-induced cigarette price increases in Taiwan 2011-2016.

Abstract:
INTRODUCTION
This study aims to analyse the non-tax-induced price increasing strategies adopted by tobacco industry in Taiwan, a high-income country with comprehensive tobacco control policies but low tobacco taxes and a declining cigarette market.

METHODS
Using governmental tax, price and inflation data, we analysed cigarette sales volume, affordability, affordability elasticity of demand, market share, pricing and net revenue of the top five tobacco companies in Taiwan from 2011 to 2016 when no tax increases occurred.

RESULTS
Total revenue after tax grew significantly for all the major transnational tobacco companies between 2011 and 2016 at the expense of the state-owned Taiwan Tobacco and Liquor Corporation. In terms of market share, Japan Tobacco (JT) was the leading company, despite experiencing a small decline, while British American Tobacco and Imperial Brands remained stable, and Philip Morris International increased from 4.7% to 7.0%. JT adopted the most effective pricing strategy by increasing the real price of its two most popular brands (Mevius and Mi-Ne) and, at the same time, doubling the sales of its cheaper and less popular brand Winston by leaving its nominal retail price unaltered.

CONCLUSIONS
Low and unchanged tobacco taxes enable tobacco companies to use aggressive pricing and segmentation strategies to increase the real price of cigarettes without making them less affordable while simultaneously maintaining customers' loyalty. It is crucial to continue monitoring the industry's pricing strategies and to regularly increase taxes to promote public health and to prevent tobacco industry from profiting at the expense of government revenues.


Clinical outcomes of endoscopic resection for the treatment of gastric gastrointestinal stromal tumors originating from the muscularis propria: a 7-year experience from a large tertiary center in China.

Abstract:
BACKGROUND AND AIMS
Few studies have evaluated the value of endoscopic resection (ER) for the treatment of gastric gastrointestinal (GI) stromal tumors (GISTs) originating from the muscularis propria (MP) in a large population, and no studies have evaluated risk factors for piecemeal resection. This study aimed to evaluate the efficacy and safety of ER for gastric GISTs in a real-world setting and to explore factors associated with piecemeal resection.

METHODS
From January 2013 to December 2019, 185 patients with gastric GISTs originating from the MP were assessed. Clinicopathological and endoscopic data were collected and analyzed. Baseline characteristics of the en bloc resection and piecemeal resection groups were compared to evaluate predictive factors for piecemeal resection.

RESULTS
There were 71 males and 114 females with a mean age of 57.0 ± 9.7 years. The mean size of GISTs was 15 mm (range 4-65 mm). A total of 123 were very low risk, 52 were low risk, and ten were moderate risk. In this study, 103 GISTs were treated with endoscopic submucosal excavation (ESE), 68 GISTs were treated with endoscopic full-thickness resection (EFR), and 14 GISTs were treated with submucosal tunneling ER (STER). Either en bloc resection or complete resection was achieved in 160 (86.5%) patients. No recurrence was noted during follow-up. Only five patients experienced minor complications, with a complication rate of 2.7%. Multivariate analysis demonstrated that size (odds ratio [OR] 1.060, 95% confidence interval [CI] 1.004-1.118; P = 0.035) and shape (OR 5.434, 95% CI 1.638-18.027; P = 0.006) were independent predictors of piecemeal resection.

CONCLUSION
ER was effective and safe for the treatment of gastric GISTs originating from the MP. Piecemeal resection did not seem to affect the efficacy of ER, and no recurrence was noted during follow-up. Large size and irregular shape are risk factors related to piecemeal resection of ER.

Endoscopic resection
Gastric
Gastrointestinal stromal tumor
Muscularis propria


Bronchioalveolar stem cells are a main source for regeneration of distal lung epithelia in vivo.

Abstract: Bronchioalveolar stem cells (BASCs) are a potential source for lung regeneration, but direct in vivo evidence for a multipotential lineage contribution during homeostasis and disease is critically missing, since specific genetic labeling of BASCs has not been possible. We developed a novel cell tracing approach based on intein-mediated assembly of newly engineered split-effectors, allowing selective targeting of dual-marker expressing BASCs in the mouse lung. RNA sequencing of isolated BASCs demonstrates that BASCs show a distinct transcriptional profile, characterized by co-expression of bronchiolar and alveolar epithelial genes. We found that BASCs generate the majority of distal lung airway cells after bronchiolar damage but only moderately contribute to cellular turnover under homeostatic conditions. Importantly, DTA-mediated ablation of BASCs compromised proper regeneration of distal airways. The study defines BASCs as crucial components of the lung repair machinery and provides a paradigmatic example for the detection and manipulation of stem cells that cannot be recognized by a single marker alone.

D053687:Adult Stem Cells
D000818:Animals
D049109:Cell Proliferation
D002478:Cells, Cultured
D004622:Embryo, Mammalian
D057809:HEK293 Cells
D006801:Humans
D051379:Mice
D057507:Mice, 129 Strain
D008810:Mice, Inbred C57BL
D011650:Pulmonary Alveoli
D012038:Regeneration
D020545:Respiratory Mucosa

BASCs
cell tracing
lung repair
lung stem cells
split‐tTA


Media, nationalism, and the body: Exploring masculinities, male partner violence, and HIV vulnerability among female sex workers in northern Karnataka, India.

Abstract: Public health and media discourses on men and HIV prevention in India have largely focused on changing knowledge, attitudes and risk behaviour pertaining to condom use and safe sex. Little attempt has been made towards intervening in areas such as masculinity, dominant gender norms and intimate partner violence (IPV) that have been shown to have a direct link to HIV prevalence. In this paper, drawing on findings from an ethnographic study in northern Karnataka (India), we show how socio-political and communicative contexts influence and perpetuate violent behaviours by men in intimate relationships with female sex workers (FSW). We argue that constructions of masculinity, the stereotypes of which are reinforced through contemporary media, and movies, are intricately linked with processes of nationalism and play out in forms of chauvinism among working-class men. Violence, celebrated through various patriarchal discourses, legitimises and reinforces gender ideals that govern the private lives of men and their female intimate partners. This study provides a complex and nuanced understanding of structural factors that lead to IPV against FSWs and offers implications for HIV intervention planning in the region and beyond.

Khan|Shamshad|S| Nair|Sapna|S| Huynh|Anthony|A| Chevrier|Claudyne|C| Thalinja|Raghavendra|R| Prakash|Ravi|R| Javalkar|Prakash|P| Ramanaik|Satyanarayana|S| Isac|Shajy|S| Bhattacharjee|Parinita|P| Lorway|Robert|R|

Masculinity
female sex workers
intimate partner violence
HIV/AIDS
health communication


Sex differences in rates of obesity in bipolar disorder: postulated mechanisms.

Abstract:
OBJECTIVE
The increased standardized mortality ratio (SMR) from cardiovascular disease (CVD) in women with bipolar disorder (BD), relative to men with BD and individuals of both sexes in the general population, provides the impetus to identify factors that contribute to the differential association of obesity with BD in women.

METHODS
We conducted a selective PubMed search of English-language articles published from September 1990 to June 2012. The key search terms were bipolar disorder and metabolic syndrome cross-referenced with gender, sex, obesity, diabetes mellitus, hypertension, and dyslipidemia. The search was supplemented with a manual review of relevant article reference lists. Articles selected for review were based on author consensus, the use of a standardized experimental procedure, validated assessment measures, and overall manuscript quality.

RESULTS
It is amply documented that adults with BD are affected by the metabolic syndrome at a rate higher than the general population. Women with BD, when compared to men with BD and individuals of both sexes in the general population, have higher rates of abdominal obesity. The course and clinical presentation of BD manifest differently in men and women, wherein women exhibit a higher frequency of depression predominant illness, a later onset of BD, more seasonal variations in mood disturbance, and increased susceptibility to relapse. Phenomenological factors can be expanded to include differences in patterns of comorbidity between the sexes among patients with BD. Other factors that contribute to the increased risk for abdominal obesity in female individuals with BD include reproductive life events, anamnestic (e.g., sexual and/or physical abuse), lifestyle, and iatrogenic.

CONCLUSIONS
A confluence of factors broadly categorized as broad- and sex-based subserve the increased rate of obesity in women with BD. It remains a testable hypothesis that the increased abdominal obesity in women with BD mediates the increased SMR from CVD. A clinical recommendation that emerges from this review is amplified attention to the appearance, or history, of factors that conspire to increase obesity in female patients with BD.

Baskaran|Anusha|A|
Cha|Danielle S|DS|
Powell|Alissa M|AM|
Jalil|Dalya|D|
McIntyre|Roger S|RS|

obesity
bipolar disorder
women
metabolic syndrome
female


Nanoscale topographic changes on sterilized glass surfaces affect cell adhesion and spreading.

Abstract: Producing sterile glass surfaces is of great importance for a wide range of laboratory and medical applications, including in vitro cell culture and tissue engineering. However, sterilization may change the surface properties of glass and thereby affect its use for medical applications, for instance as a substrate for culturing cells. To investigate potential effects of sterilization on glass surface topography, borosilicate glass coverslips were left untreated or subjected to several common sterilization procedures, including low-temperature plasma gas, gamma irradiation and steam. Imaging by atomic force microscopy demonstrated that the surface of untreated borosilicate coverslips features a complex landscape of microislands ranging from 1000 to 3000 nm in diameter and 1 to 3 nm in height. Steam treatment completely removes these microislands, producing a nanosmooth glass surface. In contrast, plasma treatment partially degrades the microisland structure, while gamma irradiation has no effect on microisland topography. To test for possible effects of the nanotopographic structures on cell adhesion, human gingival fibroblasts were seeded on untreated or sterilized glass surfaces. Analyzing fibroblast adhesion 3, 6, and 24 h after cell seeding revealed significant differences in cell attachment and spreading depending on the sterilization method applied. Furthermore, single-cell force spectroscopy revealed a connection between the nanotopographic landscape of glass and the formation of cellular adhesion forces, indicating that fibroblasts generally adhere weakly to nanosmooth but strongly to nanorough glass surfaces. Nanotopographic changes induced by different sterilization methods may therefore need to be considered when preparing sterile glass surfaces for cell culture or biomedical applications.

Wittenburg|Gretel|G|
Lauer|Günter|G|
Oswald|Steffen|S|
Labudde|Dirk|D|
Franz|Clemens M|CM|

cell adhesion
single-cell force spectroscopy
AFM
glass sterilization
nanotopography


Spirometric Reference Equations for Elderly Chinese in Jinan Aged 60-84 Years.

Abstract:
Background
The interpretation of spirometry varies on different reference values. Older people are usually underrepresented in published predictive values. This study aimed at developing spirometric reference equations for elderly Chinese in Jinan aged 60-84 years and to compare them to previous equations.

Methods
The project covered all of Jinan city, and the recruitment period lasted 9 months from January 1, 2017 to September 30, 2017, 434 healthy people aged 60-84 years who had never smoked (226 females and 208 males) were recruited to undergo spirometry. Vital capacity (VC), forced VC (FVC), forced expiratory volume in 1 s (FEV1), FEV1/FVC, FEV1/VC, FEV6, peak expiratory flow, and forced expiratory flow at 25%, 50%, 75%, and 25-75% of FVC exhaled (FEF25%, FEF50%, FEF75%, and FEF25-75%) were analyzed. Reference equations for mean and the lower limit of normal (LLN) were derived using the lambda-mu-sigma method. Comparisons between new and previous equations were performed by paired t-test.

Results
New reference equations were developed from the sample. The LLN of FEV1/FVC, FEF25-75%computed using the 2012-Global Lung Function Initiative (GLI) and 2006-Hong Kong equations were both lower than the new equations. The biggest degree of difference for FEV1/FVC was 19% (70.46% vs. 59.29%, t = 33.954, P < 0.01) and for maximal midexpiratory flow (MMEF, equals to FEF25-75%) was 22% (0.82 vs. 0.67, t = 21.303, P < 0.01). The 1990-North China and 2009-North China equations predicted higher mean values of FEV1/FVC and FEF25-75%than the present model. The biggest degrees of difference were -4% (78.31% vs. 81.27%, t = -85.359, P < 0.01) and -60% (2.11 vs. 4.68, t = -170.287, P < 0.01), respectively.

Conclusions
The newly developed spirometric reference equations are applicable to elderly Chinese in Jinan. The 2012-GLI and 2006-Hong Kong equations may lead to missed diagnoses of obstructive ventilatory defects and the small airway dysfunction, while traditional linear equations for all ages may lead to overdiagnosis.

Tian|Xin-Yu|XY| Liu|Chun-Hong|CH| Wang|De-Xiang|DX| Ji|Xiu-Li|XL| Shi|Hui|H| Zheng|Chun-Yan|CY| Xie|Meng-Shuang|MS| Xiao|Wei|W|

Aged
Spirometry
Reference Values
Respiratory Function Tests


The carboxyl tail of connexin32 regulates gap junction assembly in human prostate and pancreatic cancer cells.

Abstract: Connexins, the constituent proteins of gap junctions, are transmembrane proteins. A connexin (Cx) traverses the membrane four times and has one intracellular and two extracellular loops with the amino and carboxyl termini facing the cytoplasm. The transmembrane and the extracellular loop domains are highly conserved among different Cxs, whereas the carboxyl termini, often called the cytoplasmic tails, are highly divergent. We have explored the role of the cytoplasmic tail of Cx32, a Cx expressed in polarized and differentiated cells, in regulating gap junction assembly. Our results demonstrate that compared with the full-length Cx32, the cytoplasmic tail-deleted Cx32 is assembled into small gap junctions in human pancreatic and prostatic cancer cells. Our results further document that the expression of the full-length Cx32 in cells, which express the tail-deleted Cx32, increases the size of gap junctions, whereas the expression of the tail-deleted Cx32 in cells, which express the full-length Cx32, has the opposite effect. Moreover, we show that the tail is required for the clustering of cell-cell channels and that in cells expressing the tail-deleted Cx32, the expression of cell surface-targeted cytoplasmic tail alone is sufficient to enhance the size of gap junctions. Our live-cell imaging data further demonstrate that gap junctions formed of the tail-deleted Cx32 are highly mobile compared with those formed of full-length Cx32. Our results suggest that the cytoplasmic tail of Cx32 is not required to initiate the assembly of gap junctions but for their subsequent growth and stability. Our findings suggest that the cytoplasmic tail of Cx32 may be involved in regulating the permeability of gap junctions by regulating their size.

Katoch|Parul|P|
Mitra|Shalini|S|
Ray|Anuttoma|A|
Kelsey|Linda|L|
Roberts|Brett J|BJ|
Wahl|James K|JK|
Johnson|Keith R|KR|
Mehta|Parmender P|PP|

D015972:Gene Expression Regulation, Neoplastic

Gap Junction
Gap Junction
Connexin
Cell Adhesion
Cell junction
Gap Junctions
Protein Assembly


Search for schizophrenia and bipolar biotypes using functional network properties.

Abstract:
INTRODUCTION
Recent studies support the identification of valid subtypes within schizophrenia and bipolar disorder using cluster analysis. Our aim was to identify meaningful biotypes of psychosis based on network properties of the electroencephalogram. We hypothesized that these parameters would be more altered in a subgroup of patients also characterized by more severe deficits in other clinical, cognitive, and biological measurements.

METHODS
A clustering analysis was performed using the electroencephalogram-based network parameters derived from graph-theory obtained during a P300 task of 137 schizophrenia (of them, 35 first episodes) and 46 bipolar patients. Both prestimulus and modulation of the electroencephalogram were included in the analysis. Demographic, clinical, cognitive, structural cerebral data, and the modulation of the spectral entropy of the electroencephalogram were compared between clusters. Data from 158 healthy controls were included for further comparisons.

RESULTS
We identified two clusters of patients. One cluster presented higher prestimulus connectivity strength, clustering coefficient, path-length, and lower small-world index compared to controls. The modulation of clustering coefficient and path-length parameters was smaller in the former cluster, which also showed an altered structural connectivity network and a widespread cortical thinning. The other cluster of patients did not show significant differences with controls in the functional network properties. No significant differences were found between patients´ clusters in first episodes and bipolar proportions, symptoms scores, cognitive performance, or spectral entropy modulation.

CONCLUSION
These data support the existence of a subgroup within psychosis with altered global properties of functional and structural connectivity.

schizophrenia
biotypes
network
bipolar disorder
electroencephalogram
diffusion


Aortic stiffness increases in proportion to the severity of apnoea-hypopnea index in patients with obstructive sleep apnoea syndrome.

Abstract:
BACKGROUND AND AIMS
Obstructive sleep apnoea syndrome (OSA) and aortic stiffness are associated with an increased risk of cardiovascular morbidity and mortality. Although aortic stiffness increased in patients with OSA, the relationship between severity of OSA indicated with apnoea-hypopnea index (AHI) and aortic stiffness was not investigated in previous studies. The aim of this study is to investigate the relationship between the severity of OSA and aortic stiffness.

METHODS
In the present study, 90 consecutive OSA patients definite diagnosed with sleep test were prospectively included (mean age 54.5 ± 11.6 years). Aortic pulse wave velocity (PWV) and augmentation index (AIx) were calculated using the single-point method via the Mobil-O-Graph® ARCsolver algorithm. Aortic distensibility (AD) was calculated from the echocardiographically derived ascending aorta diameters and haemodynamic pressure measurements. Overnight full-laboratory polysomnography examination was conducted on each subject. Patients were classified into two groups according to their median AHI values (AHIlow and AHIhigh groups).

RESULTS
PWV values were higher and AD values were lower in AHIhigh group compared with AHIlow group (P < 0.05, for all). AHI was associated with body mass index (BMI), systolic blood pressure, pulse pressure, aortic diameter, AD, AIx and PWV in bivariate analysis (P < 0.05, for all). Multivariate linear regression analysis showed that AHI was independently associated with BMI (β = 0.175, P = 0.047), PWV (β = 0.521, P < 0.001) and aortic distensibility (β = -0.223, P = 0.020).

CONCLUSIONS
Aortic stiffness is associated both with the presence and the severity of OSA.

Çörtük|Mustafa|M|
Akyol|Selahattin|S|
Baykan|Ahmet O|AO|
Kiraz|Kemal|K|
Uçar|Hakan|H|
Çaylı|Murat|M|
Kandiş|Hayati|H|

obstructive sleep apnoea
pulse wave velocity
aorta
vascular stiffness


Analgesic and anti-inflammatory controlled-released injectable microemulsion: Pseudo-ternary phase diagrams, in vitro, ex vivo and in vivo evaluation.

Abstract:
PURPOSE
Development of analgesic and anti-inflammatory controlled-released injectable microemulsions utilising lysine clonixinate (LC) as model drug and generally regarded as safe (GRAS) excipients.

METHODS
Different microemulsions were optimised through pseudo-ternary phase diagrams and characterised measuring droplet size, viscosity, ex vivo haemolytic activity and in vitro drug release. The anti-inflammatory and analgesic activity was tested in mice (Hot plate test) and rats (Carrageenan-induced paw edema test) respectively and their activity was compared to an aqueous solution of LC salt.

RESULTS
The aqueous solution showed a faster and shorter response whereas the optimised microemulsion increased significantly (p<0.01) the potency and duration of the analgesic and anti-inflammatory activity after deep intramuscular injection. The droplet size and the viscosity were key factors to control the drug release from the systems and enhance the effect of the formulations.

CONCLUSIONS
The microemulsion consisting of Labrafil®/Lauroglycol®/Polysorbate 80/water with LC (56.25/18.75/15/10, w/w) could be a promising formulation after buccal surgery due to its ability to control the drug release and significantly achieve greater analgesic and anti-inflammatory effect over 24h.

Pineros|Isabel|I|
Slowing|Karla|K|
Serrano|Dolores R|DR|
de Pablo|Esther|E|
Ballesteros|Maria Paloma|MP|

Analgesic
Anti-inflammatory
Controlled-release
Injectable
Microemulsion
Lysine clonixinate
Depot formulation
Pseudo-ternary diagrams


The role of ERK2 in colorectal carcinogenesis is partly regulated by TRAPPC4.

Abstract: The extracellular signal-regulated kinase (ERK), mitogen-activated protein kinase (MAPK) pathway is an important cell proliferation pathway. We previously reported that the transport protein particle complex 4 (TRAPPC4), ERK2 interaction may activate ERK1/2, modulate pERK2 nuclear localization and regulate proliferation and apoptosis in colorectal cancer (CRC) cells. The present study further investigated the function of the TRAPPC4-ERK2 interaction in CRC in vitro and in vivo. Silencing of TRAPPC4 induced G0/G1 phase cell cycle arrest, upregulated p21 and downregulated cyclin B1 in CRC cells. Overexpression of TRAPPC4 after ERK2 silencing decreased the percentage of G0/G1 phase cells, increased the percentage of G2/M and S phase cells, downregulated p21, upregulated cyclin B1, and enhanced CRC cell viability. Immunohistochemical staining revealed that knockdown of TRAPPC4 downregulated pERK2, whereas overexpression of TRAPPC4 upregulated pERK2. Epidermal growth factor (EGF) stimulated upregulation of TRAPPC4 and pERK2 in SW1116 cells; EGF stimulation or overexpression of TRAPPC4 induced pERK2 nuclear translocation. Silencing of TRAPPC4 reduced SW1116 xenograft tumor growth in vivo, whereas overexpression of TRAPPC4 increased tumor growth, compared to control tumors. Moreover, modulation of TRAPPC4 expression in vivo affected the levels of pERK2 in the cytoplasm and nucleus and expression of p21. These results conclusively demonstrate that TRAPPC4 regulates ERK2 activation and also affects the distribution of activated pERK2 in CRC cells. The ability of ERK2 to play a role in colorectal carcinogenesis depends, at least in part, on TRAPPC4.

Weng|Yu-Rong|YR| Kong|Xuan|X| Yu|Ya-Nan|YN| Wang|Ying-Chao|YC| Hong|Jie|J| Zhao|Shu-Liang|SL| Fang|Jing-Yuan|JY|

D019950:Mitogen-Activated Protein Kinase 1

D048052:Mitogen-Activated Protein Kinase 3


Dairy and Growth, Latest Findings, and Lessons Learned.

Abstract:
BACKGROUND
Low birth weight is a significant risk factor in infant mortality and morbidity and the likelihood of chronic and clinical disease in later life.

OBJECTIVE
To identify reported associations between maternal dietary patterns and risk of low birth weight offspring.

METHODS
This article reviews some of the literature on this subject, with emphasis on maternal intake of dairy products.

RESULTS
Published data show a positive association between maternal dairy product intake and birth weight but are inconclusive with respect to fetal length. While data are limited, 2 studies have reported a positive association between maternal consumption of milk and yogurt and birth weight, but this association is not observed with respect to consumption of cheese.

CONCLUSION
If correct, by deduction, this observation is suggestive that whey protein may be a contributing factor to birth weight. Indirect evidence supporting why this observation merits further investigation is presented.

D039382:Maternal Nutritional Physiological Phenomena

low birth weight
whey protein
maternal nutrition
pregnancy


Cross-Cultural Adaptation, Reliability, and Psychophysical Validation of the Pain and Sleep Questionnaire Three-Item Index in Finnish.

Abstract: Reciprocal relationships between chronic musculoskeletal pain and various sleep disturbances are well established. The Pain and Sleep Questionnaire three-item index (PSQ-3) is a concise, valid, and reliable patient-reported outcome measure (PROM) that directly evaluates how sleep is affected by chronic low back pain (CLBP). Translation and cross-cultural validation of The Pain and Sleep Questionnaire three-item index Finnish version (PSQ-3-FI) were conducted according to established guidelines. The validation sample was 229 subjects, including 42 pain-free controls and 187 subjects with chronic musculoskeletal pain. Our aims were to evaluate internal consistency, test-retest reliability, measurement error, structural validity, convergent validity, and discriminative validity and, furthermore, to study the relationships between dizziness, postural control on a force plate, and objective sleep quality metrics and total PSQ-3-FI score. The PSQ-3-FI demonstrated good internal consistency, excellent test-retest reliability, and small measurement error. Confirmatory factor analysis confirmed acceptable fit indices to a one-factor model. Convergent validity indicated fair to good correlation with pain history and well-established pain-related PROMs. The PSQ-3-FI total score successfully distinguished between the groups with no pain, single-site pain, and multisite pain. A higher prevalence of dizziness, more impaired postural control, and a general trend towards poorer sleep quality were observed among subjects with higher PSQ-3-FI scores. Postural control instability was more evident in eyes-open tests. The Finnish PSQ-3 translation was successfully cross-culturally adapted and validated. The PSQ-3-FI appears to be a valid and reliable PROM for the Finnish-speaking CLBP population. More widespread implementation of PSQ-3 would lead to better understanding of the direct effects of pain on sleep.

Mikkonen|Jani|J|0000-0001-5308-9182 Leinonen|Ville|V|0000-0002-4282-4687 Luomajoki|Hannu|H| Kaski|Diego|D| Kupari|Saana|S| Tarvainen|Mika|M| Selander|Tuomas|T| Airaksinen|Olavi|O|

pain
sleep
questionnaire
patient-reported outcome measure
chronic low back pain
cross-cultural validation
dizziness
postural control
sleep quality
actigraphy


Monitoring seasonal distribution of an endangered anadromous sturgeon in a large river using environmental DNA.

Abstract: Monitoring dynamic distribution is crucial to conservation management of anadromous sturgeons, but traditional survey methods are less efficient for low-density populations in a large river. Natural propagation of Chinese sturgeon has been monitored annually mainly at the spawning ground using netting for eggs and hydroacoustics for broodstock. However, absence of spawning was observed sporadically in recent years, indicating further crises for the declining population. We analyzed eDNA of water samples collected from 24 sites across 1360 km of the migratory route of anadromous Chinese sturgeon in the middle and lower reaches of the Yangtze River. Chinese sturgeon was detected at 9 sites during the spawning season and 14 sites after the spawning season. We found that positive eDNA detection rates remained constant in the middle reaches but dramatically changed in the lower reaches, reflecting seasonal migration pattern of Chinese sturgeon. Invasive sturgeons were detected in the river, indicating their possible escape from aquaculture facilities. This study established a protocol for the use of eDNA to monitor distribution of Chinese sturgeon and could be valuable in making better policies for the conservation of this endangered species.

Xu|Nian|N| Zhu|Bin|B| Shi|Fang|F| Shao|Ke|K| Que|Yanfu|Y| Li|Weitao|W| Li|Wei|W| Jiao|Wenjing|W| Tian|Hua|H| Xu|Dongmei|D| Chang|Jianbo|J|

D003247:Conservation of Natural Resources

Chinese sturgeon
eDNA
Yangtze River
Conservation program
Spawning migration
Species detection


Outcome evaluation of the opioid agonist maintenance treatment in Iran.

Abstract:
INTRODUCTION AND AIMS
Methadone maintenance treatment and buprenorphine maintenance treatment are the two main therapeutic options considered for opioid replacement therapy. This study was conducted to examine the effectiveness of methadone maintenance treatment and buprenorphine maintenance treatment in Iran under usual clinical conditions.

DESIGN AND METHODS
In this outcome research, 311 patients consented to participate in the study (77.8% response rate). The Opioid Treatment Index, General Health Questionnaire and WHOQOL-BREF questionnaire were used to assess the effectiveness of the therapeutic programs. Drop-out rate was calculated after two and six months of treatment.

RESULTS
Mean dose of methadone was in an acceptable range; however, doses for buprenorphine maintenance treatment patients seemed low. The rates of attrition after two and six months of treatment were 24.2% and 44.0% in the methadone maintenance treatment group and 41.3% and 65.4% in the buprenorphine maintenance treatment group, respectively (P < 0.001). Drug use, HIV risk-taking behaviour, and mental and physical health improved in both groups at six months of treatment, while quality of life improved only in the methadone maintenance treatment group.

DISCUSSION AND CONCLUSIONS
The retention seen in the buprenorphine group may in part be related to the low buprenorphine doses used. As a whole, the positive results provide support to continuation of maintenance programs.

Esmaeili|Hamid-Reza|HR| Ziaddinni|Hassan|H| Nikravesh|Mohammad-Rafee|MR| Baneshi|Mohammad-Reza|MR| Nakhaee|Nouzar|N|

patient dropout/statistics and numerical data


Volunteers trained in palliative care at the hospital: an original and dynamic resource.

Abstract:
OBJECTIVE
Volunteers trained in palliative care are increasingly present in acute care units in general hospitals. Nevertheless, there still are few available data on this topic, especially concerning the integration of volunteers outside the palliative structures. Our present study aimed to describe the experience of volunteers trained in palliative care in the context of a primary care hospital. In particular, the difficulties and the benefits of this specific position were evaluated according to volunteers' own perceptions and words.

METHOD
We employed a qualitative method. Various aspects of the volunteer's role were explored by means of semistructured questions, addressing their activity, their motivations, and their feelings. Participants were volunteers (n = 19) trained in palliative care and working at a university hospital. After giving written consent, they completed the semistructured questionnaire at home. Content analysis was used to identify the main categories of answers and the principal themes reported by the volunteers.

RESULTS
The main difficulties were related to uncertainty of the context. As every situation is different, volunteers could not define their role once and for all. However, they derived great satisfaction from their activity. A supporting frame and a good balance between constraints and autonomy were facilitating factors. Besides, the complexity related to the context contributed to make the position valuable and challenging.

SIGNIFICANCE OF RESULTS
Integrating a voluntary service in a primary care hospital is partly based on active participation of the volunteers in developing their position in a more adequate way. In return, this relative autonomy implies a rigorous and supportive attitude from the institution.

Delaloye|Sylvette|S| Escher|Monica|M| Luthy|Christophe|C| Piguet|Valérie|V| Dayer|Pierre|P| Cedraschi|Christine|C|

General hospitals
Palliative and supportive care
Palliative care volunteers
Role identification
Volunteer satisfaction


How do tendons adapt? Going beyond tissue responses to understand positive adaptation and pathology development: A narrative review.

Abstract: Understanding how tendons adapt to load is crucial to understanding how training can improve performance, minimise the risk of injury and aid rehabilitation. Adaptation is the positive response of an organism or tissue to benefit its function. In tendons, numerous tissue responses to load have been identified in vivo. Changes in tendon dimensions, structure on imaging, mechanical properties, and blood flow have been reported in response to mechanical stimuli. However, research has focused on tissue level changes with little understanding of how changes at the tissue level affect the person, their athletic performance or injury risk. Tendons can have a paradoxical response to load, load can induce positive adaptation, however it is also a major factor in the development of tendon pathology and pain. Tendon pathology is a risk factor for developing symptoms, yet the high rate of asymptomatic pathology suggests that the pathological tendon must adapt to be able to tolerate load. Similarly, there is mounting evidence to suggest that tendon remodelling or repair is not necessary for a positive clinical outcome following rehabilitation, suggesting that the tendon must adapt via other mechanisms. This narrative review synthesises evidence of how normal and pathological tendons adapts to load, and how this relates to adaptation of load capacity and function of the individual.

Tendon Adaptation Pathology Injury Rehabilitation


Realizing Crack Diagnosing and Self-Healing by Electricity with a Dynamic Crosslinked Flexible Polyurethane Composite.

Abstract: Combining self-healing functions with damage diagnosing, which can achieve timely healing autonomously, is expected to improve the reliability and reduce life cycle cost of materials. Here, a flexible conductive composite composed of a dynamically crosslinked polyurethane bearing Diels-Alder bonds (PUDA) and carbon nanotubes (CNTs), which possess both crack diagnosing and self-healing functions, is reported. The introduced dynamic Diels-Alder bonds endow the materials self-healing function and the powder-based preparation route based on the specially designed CNTs-coated PUDA micropowders leads to the formation of segregated CNTs network, which makes the composite possess excellent mechanical properties and high conductivity. Because of the sufficient electrothermal and photothermal effect of CNTs, the composites can be healed rapidly and repeatedly by electricity or near-infrared light based on the retro-Diels-Alder reaction. An obvious color difference in the infrared thermograph resulting from the resistance difference between damaged and undamaged area can be observed when applying the voltage, which can be used for crack diagnosing. Using the same electrical circuit, the crack in the PUDA/CNTs composite can be noninvasively detected first and then be autonomously healed. The composites also exhibit a strain-sensing function with good sensitivity and high reliability, thus will have potential applications in electronic strain sensors.

Pu|Wuli|W| Fu|Daihua|D| Wang|Zhanhua|Z| Gan|Xinpeng|X| Lu|Xili|X| Yang|Li|L| Xia|Hesheng|H|0000-0003-2309-1403

crack diagnosing
polyurethane
carbon nanotubes
self‐healing


Designing and Construction of a Cloning Vector Containing mpt64 Gene of Mycobacterium tuberculosis.

Abstract:
Background
Tuberculosis caused by Mycobacterium tuberculosis (M. tuberculosis), remains as one of the leading causes of deaths worldwide, with nearly two million death cases annually. BCG (Bacille Calmette-Guerin) continues to be the most widely used vaccine in the world, but the protective immunity differs in different parts of the world. Accordingly, new strategies including DNA vaccines are essentially needed. This study was aimed to design and construct a cloning vector containing mpt64 gene of M. tuberculosis.

Materials and Methods
M. tuberculosis H37Rv was cultured on Lowenstein Jensen medium, and genomic DNA was extracted. The mpt64 gene was amplified by PCR using designed specific primers. After the digestion of mpt64 and pcDNA3.1 (+) by BamHI and EcoRI restriction enzymes, the mpt64 fragment was ligated into the digested vector using T4 DNA ligase enzyme. Then, the recombinant vector was transformed into competent Escherichia coli (E. coli) TOP10 strain. To confirm the colonies of transformed bacteria, antibiotic resistance, colony-PCR, restriction enzyme digestion and DNA sequencing were used.

Results
To confirm the clones, colony-PCR using mpt64 specific primers was performed and the fragment of 718 bp was observed by gel electrophoresis. Clones were also verified by restriction enzyme digestion using BamHI and EcoRI restriction enzymes and the 718 bp fragment was observed. Furthermore, results of DNA sequencing showed 100% homology with the mpt64 fragment of H37Rv in GenBank.

Conclusion
In this study, the mpt64 fragment was successfully cloned in pcDNA3.1 (+) vector. This construct can be used in future studies as a DNA vaccine in animal models to induce immune system responses.

Zare|Hosna|H| Aryan|Ehsan|E| Alami|Shadi|S| Yaghoubi|Atieh|A| Teimourpour|Roghayeh|R| Meshkat|Zahra|Z|

cloning vector
mpt64
Mycobacterium tuberculosis
DNA vaccine


Dynamic role and importance of surrogate species for assessing potential adverse environmental impacts of genetically engineered insect-resistant plants on non-target organisms.

Abstract: Surrogate species have a long history of use in research and regulatory settings to understand the potentially harmful effects of toxic substances including pesticides. More recently, surrogate species have been used to evaluate the potential effects of proteins contained in genetically engineered insect resistant (GEIR) crops. Species commonly used in GEIR crop testing include beneficial organisms such as honeybees, arthropod predators, and parasitoids. The choice of appropriate surrogates is influenced by scientific factors such as the knowledge of the mode of action and the spectrum of activity as well as societal factors such as protection goals that assign value to certain ecosystem services such as pollination or pest control. The primary reasons for using surrogates include the inability to test all possible organisms, the restrictions on using certain organisms in testing (e.g., rare, threatened, or endangered species), and the ability to achieve greater sensitivity and statistical power by using laboratory testing of certain species. The acceptance of surrogate species data can allow results from one region to be applied or "transported" for use in another region. On the basis of over a decade of using surrogate species to evaluate potential effects of GEIR crops, it appears that the current surrogates have worked well to predict effects of GEIR crops that have been developed (Carstens et al. GM Crops Food 5:1-5, 2014), and it is expected that they should work well to predict effects of future GEIR crops based on similar technologies.

Wach|Michael|M|0000-0001-7151-0727
Hellmich|Richard L|RL|
Layton|Raymond|R|
Romeis|Jörg|J|
Gadaleta|Patricia G|PG|

Genetically engineered insect resistance


Towards rectifying limitations on species delineation in dusky salamanders (Desmognathus: Plethodontidae): An ecoregion-drainage sampling grid reveals additional cryptic clades.

Abstract: Dusky salamanders (Desmognathus) constitute a large, species-rich group within the family Plethodontidae, and though their systematic relationships have been addressed extensively, most studies have centered on particular species complexes and therefore offer only piecemeal phylogenetic perspective on the genus. Recent work has revealed Desmognathus to be far more clade rich-35 reciprocally monophyletic clades versus 22 recognized species-than previously imagined, results that, in turn, provide impetus for additional survey effort within clades and across geographic areas thus far sparsely sampled. We conceived and implemented a sampling regime combining level IV ecoregions and independent river drainages to yield a geographic grid for comprehensive recovery of all genealogically exclusive clades. We sampled over 550 populations throughout the distribution of Desmognathus in the eastern United States of America and generated mitochondrial DNA sequence data (mtDNA; 1,991 bp) for 536 specimens. A Bayesian phylogenetic reconstruction of the resulting haplotypes revealed forty-five reciprocally monophyletic clades, eleven of which have never been included in a comprehensive phylogenetic reconstruction, and an additional three not represented in any molecular systematic survey. Although general limitations associated with mtDNA data preclude new species delineation, we profile each of the 45 clades and assign names to 10 new clades (following a protocol for previous clade nomenclature). We also redefine several species complexes and erect new informal species complexes. Our dataset, which contains topotypic samples for nearly every currently recognized species and most synonymies, will offer a robust framework for future efforts to delimit species within Desmognathus.

Amphibia, Caudata, Desmognathus, mtDNA phylogeny, level IV ecoregion X independent drainage sampling, new clades


Simalikalactone D, a Potential Anticancer Compound from Simarouba tulae, an Endemic Plant of Puerto Rico.

Abstract: Species of the genus Simarouba have been studied because of their antimalarial and antileukemic activities. A group of oxygenated terpenes called quassinoids have been isolated from species of the Simarouba genus, and are responsible for its therapeutic properties. We hypothesized that Simarouba tulae, an endemic plant from Puerto Rico, is a natural source rich in quassinoid compounds with anticancer activity. The leaves were processed and extracted with solvents of different polarities. The extracts were screened for their antiproliferative activity, and it was shown that the chloroform extract was the most active extract. This extract was purified using different chromatographic techniques to afford the quassinoid simalikalactone D (SKD). This compound was further characterized using NMR and X-ray diffraction analysis. A reassessment of original structural assignments for SKD is proposed. SKD showed high cytotoxicity activity, with an IC50 of 55, 58, and 65 nM in A2780CP20 (ovarian), MDA-MB-435 (breast), and MDA-MB-231 (breast) cell lines, respectively. Exposure to SKD led to 15% inhibition of the migration of MDA-MB-231 cells.

simalikalactone D
Simarouba tulae
quassinoids
migration
breast cancer
medicinal plants
ovarian cancer


The Role of Environmental Health Literacy When Developing Traffic Pollution Fact Sheets for Puerto Rican Adults.

Abstract: Environmental health literacy is particularly relevant to racial/ethnic and linguistic minority populations who are likely to live near major roadways and highways. We conducted exploratory research to develop ways to communicate the risks of traffic-related air pollution to Puerto Rican adults living in and near Boston, Massachusetts. We held two initial focus groups with Spanish-speaking Puerto Rican adults (N = 16) enrolled in the Boston Puerto Rican Health Study (BPRHS). Most had only a high school education or less and earned a low income. We used thematic analysis of transcripts to suggest ways to improve three fact sheets designed to communicate BPRHS findings to the community. Based on these results, we made substantial revisions. We then conducted a second set of two focus groups with the same participants to assess revisions. Participants viewed the revised fact sheets more favorably and indicated that they found them easier to read. Lessons learned about improving readability and relevance included increasing text size, adding more graphics, chunking text, and providing more technical details. Designing successful environmental health communication tools that retain scientific accuracy is not a simple task. There is need for systematic attempts to evaluate and report on health literacy and community engagement processes for developing materials that are easy to read, culturally relevant, and that communicate complex environmental health information and concepts in ways people can understand and act on.

Brugge|Doug|D| Tracy|Migdalia|M| Thayer|Katherine|K| Thayer|Anne|A| Dayer|Brianna|B| Figueroa|Nancy|N| Kurtz-Rossi|Sabrina|S|

environmental health literacy
fact sheets
Puerto Rican
traffic-related air pollution
focus group
ultrafine particles


Isolating Crucial Steps in Induction of Infective Endocarditis With Preclinical Modeling of Host Pathogen Interaction.

Abstract: Animal models of Staphylococcus aureus infective endocarditis (IE), especially in rodents, are commonly used to investigate the underlying pathogenesis, disease progression, potential diagnostic approaches, and therapeutic treatment. All these models are based on surgical interventions, and imply valve trauma by placing a polyurethane catheter at the aortic root. While the influence of endothelial damage and inflammation on the induction of IE has been studied intensively, the role of the catheter, as permanent source of bacteremia, and the interplay with bacterial virulence factors during the formation of IE is poorly understood. In our study, we aimed at identifying which set of preconditions is required for induction and formation of IE: (1) tissue injury, (2) permanent presence of bacteria, and (3) presence of the full bacterial repertoire of adhesion proteins. We investigated the manifestation of the disease in different modifications of the animal model, considering different degrees of endothelial damage and the presence or absence of the catheter. In four infection models the induction of IE was assessed by using two bacterial strains with different expression patterns of virulence factors - S. aureus 6850 and Newman. In vivo magnetic resonance imaging showed conspicuous morphological structures on the aortic valves, when an endothelial damage and a continuous bacterial source were present simultaneously. Cellular and inflammatory pathophysiology were characterized additionally by histology, real-time quantitative polymerase chain reaction analysis, and bacterial counts, revealing strain-specific pathogenesis and manifestation of IE, crucially influenced by bacterial adherence and toxicity. The severity of IE was dependent on the degree of endothelial irritation. However, even severe endothelial damage in the absence of a permanent bacterial source resulted in reduced valve infection. The spread of bacteria to other organs was also dependent on the pathogenic profile of the infectious agent.

Schwarz|Christian|C|
Hoerr|Verena|V|
Töre|Yasemin|Y|
Hösker|Vanessa|V|
Hansen|Uwe|U|
Van de Vyver|Hélène|H|
Niemann|Silke|S|
Kuhlmann|Michael T|MT|
Jeibmann|Astrid|A|
Wildgruber|Moritz|M|
Faber|Cornelius|C|

endocarditis
Staphylococcus aureus
MRI
foreign body
mouse model


Clinical value of the Integrated Pulmonary Index® during sedation for interventional upper GI-endoscopy: A randomized, prospective tri-center study.

Abstract:
BACKGROUND AND STUDY AIMS
The Integrated Pulmonary Index® (IPI) is a mathematically-determined factor based on parameters of capnography and pulse oximetry, which should enable sensitive detection of impaired respiratory function. Aim was to investigate whether an additional measurement of the IPI during sedation for interventional endoscopy, compared to standard monitoring alone, allows a reduction of sedation-related respiratory depression.

PATIENTS AND METHODS
170 patients with standard monitoring randomly underwent either a blinded recording of capnography (control group, n=87) or capnography, including automated IPI calculation (IPI group, n=83), during deep sedation with midazolam and propofol. The primary endpoint was the maximum decrease of oxygen saturation from the baseline level before sedation. Secondary endpoints: incidence of hypoxemia (SaO2<90%), other sedation-related complications (apnea rate, bradycardia, hypotension), patient cooperation and satisfaction (VAS).

RESULTS
Mean propofol dose in the IPI group (245±61mg) was comparable to the control group (225±47mg). The average drop of the oxygen saturation in the IPI group (6.5±4.1%) was nearly identical to that of the control group (7.1±4.6%, p=0.44). Apnea episodes >15s was found in 46 patients of the control and 31 of the IPI group (p<0.05). Frequency of occurrence of a drop in pO2-saturation <90%, bradycardia <50/min or a drop of systolic pressure <90mmHg were not significantly different in both groups. Mechanical ventilation was not required in any case. Patient cooperation and satisfaction were assessed similar in both groups.

CONCLUSION
A clinically appealing advantage of IPI-assessment during deep sedation with midazolam and propofol for interventional endoscopy could not be documented. However, IPI registration was more effective in reducing the incidence of apnea episodes.

Riphaus|Andrea|A|
Wehrmann|Till|T|
Kronshage|Tim|T|
Geist|Christoph|C|
Pox|Christian P|CP|
Heringlake|Stefan|S|
Schmiegel|Wolff|W|
Beitz|Analena|A|
Meining|Alexander|A|
Müller|Michaela|M|
von Delius|Stefan|S|

Midazolam
Propofol
Hypoxemia
Apnea
Endoscopic resection
Endoscopic sedation
Endoscopic ultrasound
Gastrointestinal endoscopy


Potential use of membrane bioreactor to treat petroleum refinery effluent: comprehension of dynamic of organic matter removal, fouling characteristics and membrane lifetime.

Abstract: This study aims to evaluate membrane bioreactor (MBR) performance in a pilot scale to treat petroleum refinery effluent, and has been primarily focused on (1) investigation of dynamics of organic matter removal; (2) characterization of membrane fouling under real hazardous events; (3) evaluation of the effect of fouling on membrane lifetime; and (4) estimate the membrane lifetime. The results have shown that the MBR was able to effectively reduce COD, NH3-N, turbidity, color, phenol and toxicity, and bring them to the levels required to meet disposal and non-potable water reuse standards. The FTIR results showed that organic matter was removed by biological oxidation and/or retained by adsorption in the biological sludge, or retention in the UF membrane, and that SMP was produced during the treatment. In terms of membrane permeability, the results showed that soluble fraction of mixed liquor contributed significantly to membrane fouling. And finally, considering the concept of lifetime based on permeability decline, a membrane lifetime of 7 years is expected.

Alkmim|Aline R|AR|
da Costa|Paula R|PR|
Moser|Priscila B|PB|
Neta|Luzia S França|LSF|
Santiago|Vânia M J|VMJ|
Cerqueira|Ana C|AC|
Reis|Beatriz G|BG|
Amaral|Míriam C S|MCS|

D017550:Spectroscopy, Fourier Transform Infrared

Membrane bioreactor
Petroleum refinery effluent
Membrane lifetime
Membrane fouling
Water reuse


Upregulation of Vesicular Glutamate Transporter 2 and STAT3 Activation in the Spinal Cord of Mice Receiving 3,3'-Iminodipropionitrile.

Abstract: Chronic administration of 3,3'-iminodipropionitrile (IDPN) causes axonal impairment. Although controversy still remains, it has been suggested that IDPN intoxication mimics the axonopathy of amyotrophic lateral sclerosis (ALS). Interestingly, recent studies including our own showed that signal transducer and activator of transcription 3 (STAT3) in spinal α-motoneurons was activated in both IDPN-treated mice and SOD1 G93A mice, a genetic model of familial ALS. Because activation of STAT3 occurs in response to various stimuli, such as axonal injury, ischemia, and excessive glutamate, here we focused on a potential link between phosphorylated STAT3 (pSTAT3, an active form) and vesicular glutamate transporter 2 (VGluT2, a regulator of glutamate storage and release) in IDPN-treated mice and SOD1 G93A mice. Impairment of axonal transport was confirmed by western blot analysis: the expression levels of phosphorylated neurofilament H were elevated in both models. As shown in SOD1 G93A mice, the expression frequencies of VGluT2 in synaptophysin-positive (SYP)+ presynaptic terminals around spinal α-motoneurons were significantly higher in IDPN-treated mice than in vehicle controls. The coverages of spinal α-motoneurons by VGluT2+ presynaptic terminals were more elevated around pSTAT3+ cells than around pSTAT3- cells in IDPN-treated mice and SOD1 G93A mice. Considering that excessive glutamate is shown to be involved in axonal impairment and STAT3 activation, the present results suggest that IDPN-induced upregulation of VGluT2 may result in an increase in glutamate, which might cause axonopathy and induction of pSTAT3. The link between upregulation of VGluT2 and activation of STAT3 via glutamate may represent a common pathological feature of IDPN-treated mice and SOD1 G93A mice.

Ohgomori|Tomohiro|T| Yamasaki|Ryo|R| Kira|Jun-Ichi|JI| Jinno|Shozo|S|0000-0002-7596-8118

D027342:Excitatory Amino Acid Transporter 2

D050599:Vesicular Glutamate Transport Protein 2

Signal transducer and activator of transcription 3


Severe Hypercalcemia in a Postpartum Patient.

Abstract:
BACKGROUND
There are multiple clinical manifestations of hypercalcemia and several causes of hypercalcemia. Hypercalcemia caused by milk-alkali syndrome is increasing in frequency.

CASE REPORT
A 26-year-old woman presented after having undergone caesarian section. She complained of severe myalgias, arthralgias, an inability to ambulate, nausea, vomiting, abdominal pain, and marked depression. Each of these symptoms has a broad differential diagnosis, but when considered together the theme "stones, bones, moans, and groans," seen in patients with hypercalcemia, is evident. This patient was found to have hypercalcemia caused by milk-alkali syndrome related to the ingestion of calcium carbonate. Her symptoms and hypercalcemia resolved with treatment. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Emergency physicians should be aware of the many different symptoms of hypercalcemia. This case emphasizes the need for a careful medication history for any patient presenting with hypercalcemia, including over the counter medications. Physicians should have a high level of suspicion for milk-alkali syndrome in patients with hypercalcemia because milk-alkali syndrome is no longer a rare etiology but rather one of the most common causes of hypercalcemia.

Murphy|Megan L|ML|
Mattingly|Michael J|MJ|
Meisner|Craig T|CT|

D050071:Bone Density Conservation Agents

hypercalcemia
postpartum
milk-alkali syndrome
pregnancy


Characterization of a Pt mirror to be used to deflect synchrotron radiation beam onto Langmuir monolayers.

Abstract: A homemade mirror for X-rays has been built to prepare a diffraction beamline for liquid surface diffraction and scattering measurements. This simple approach is in operation at the XRD2 bending-magnet beamline at the Brazilian Synchrotron Light Laboratory.

Vieira|Carlos E P|CE|
Gasperini|Antonio A M|AA|
Freitas|Pedro P S|PP|
Oliveira|Rafael G|RG|
Cavalcanti|Leide P|LP|

Langmuir monolayer
diffraction
deflector mirror
grazing incidence


Suraksha Chakra: A new scoring system to protect the healthcare workers from COVID.

Abstract: Doctors, nurses, house cleaning staff and hospital ward attendants are at increased risk of acquiring coronavirus disease 2019 (COVID-19) if there is a breach in the personal protection equipment. A new simple, easy to implement scoring system has been developed by our Suraksha Chakra team which can be used by the policy makers and hospital administrators. The scoring system is not to discourage anyone but constantly reinforce the healthcare workers for safe practices during patient care.

Sureka|Binit|B|
Tak|Vibhor|V|
Nag|Vijaya Lakshmi|VL|
Rani|Raj|R|
Garg|Mahendra Kumar|MK|
Misra|Sanjeev|S|


Circulating irisin detection: Does it really work?

Abstract: The recent discovery of irisin has generated considerable interest in the scientific community. However, many studies on the biochemistry and biology of this intriguing hormone yielded controversial results in humans, which were mostly attributable to a number of drawbacks in the methods used for its detection and measurement.

irisin
ELISA kit
FNDC5
antibodies
exercise
mass spectrometry


Expression of inositol-requiring enzyme 1β is downregulated in colorectal cancer.

Abstract: The endoplasmic reticulum stress inositol-requiring enzyme (IRE) 1α/X-box binding protein (XBP) 1 signaling pathway is involved in the tumorigenesis of breast and prostate cancer. Mucin 2 (MUC2) protects colon tissues from the formation of tumors. In human colorectal cancer (CRC) the role of IRE1α, and its analogue, IRE1β, has yet to be elucidated. In the present study, the expression levels of IRE1α, IRE1β, un-spliced XBP1u, spliced XBP1s and MUC2 in surgically resected cancerous and adjacent non-cancerous tissues from patients with CRC were investigated. The IRE1α, IRE1β, XBP1u, XBP1s and MUC2 mRNA expression levels were determined using reverse transcription-quantitative polymerase chain reaction, and the protein expression levels were detected using immunohistochemistry and western blotting. The association between the expression levels of IRE1α, IRE1β and MUC2 and the clinicopathological features of patients with CRC was subsequently analyzed. The mRNA expression levels of IRE1β and MUC2 were decreased by ~2.1 and ~4.5-fold in CRC tissues, respectively, as compared with the adjacent normal tissues. The protein expression levels of IRE1β and MUC2 were decreased by ~8.0 and ~2.0-fold in the CRC tissues, respectively. IRE1β mRNA expression levels were positively correlated with MUC2 mRNA expression levels. IRE1β expression levels were revealed to be significantly associated with lymph node metastasis, tumor stage and histological differentiation. However, IRE1α, XBP1u and XBP1s mRNA and IRE1α protein expression levels were not observed to significantly differ between cancerous tissues and the adjacent normal tissues. The results indicated that the expression of IRE1β, but not IRE1α, may protect colon tissue from developing CRC by inducing MUC2 expression. Therefore, decreased IRE1β expression levels may be associated with the development of CRC through the inhibition of MUC2 expression.

Jiang|Yalin|Y| Zhou|Yunfeng|Y| Zheng|Yufeng|Y| Guo|Hong|H| Gao|Lei|L| Chen|Pan|P| Feng|Dandan|D| Qi|Ran|R| Li|Xiaozhen|X| Chang|Yongchao|Y| Chu|Fong-Fong|FF| Gao|Qiang|Q|

colorectal cancer
endoplasmic reticulum stress
mucin 2
X-box binding protein 1
inositol-requiring enzyme 1


Acquired urea cycle amino acid deficiency and hyperammonaemic encephalopathy in a cat with inflammatory bowel disease and chronic kidney disease.

Abstract:
Case summary
A 5-year-old male neutered Persian cat was referred for investigation of a 4 week history of weight loss, inappetence and intermittent vomiting. Chronic kidney disease (CKD) and inflammatory bowel disease were diagnosed, and despite immunosuppressive therapy and assisted enteral nutrition, the cat experienced persistent anorexia, vomiting and severe weight loss. After 2 additional weeks of treatment, the cat developed acute-onset neurological signs associated with severe hyperammonaemia and was euthanased. Plasma amino acid assessment revealed deficiency of several amino acids involved in the urea cycle, including arginine.

Relevance and novel information
To our knowledge, this is the first reported case of an acquired urea cycle amino acid deficiency without nutritional deprivation in a cat. Several contributing factors were suspected, including intestinal malabsorption and CKD. This case demonstrates the importance of urea cycle amino acids in feline metabolism and possible necessity for parenteral supplementation, particularly in the context of persistent weight loss despite adequate enteral nutrition.

amino acids
hyperammonaemic encephalopathy
inflammatory bowel disease
chronic kidney disease
hyperammonaemia
Arginine


Obstetric Paralysis: Evaluation of the Sever-L'Episcopo Technique Modified by Hoffer.

Abstract: Evaluate the results of a series of 28 cases of high obstetric paralysis treated with the Sever-L'Episcopo technique modified by Hoffer, between 2003 and 2016. Children (mean age, four years and seven months) with adduction contracture and internal rotation of the shoulder without secondary bone deformities (Mallet class II) underwent lengthening of the pectoralis major muscle and tenotomy of the subscapularis muscle associated with transfer of the latissimus dorsi and teres major muscle to the infraspinatus muscle, moving to the function of external rotators and elevators. The mean follow-up was three years and 10 months. At the end of the study, 24 patients achieved excellent functional assessment scores, mainly of the abduction and external rotation, passing from Mallet class II to class IV. Four patients still demonstrated some degree of global movement limitation, passing from class II to class III. Regardless of the final functional gain, all patients were able to perform tasks that were previously difficult. The data from this study suggest that Hoffer's surgery is an effective method in the treatment of the sequelae of high obstetric paralysis without secondary bone deformities.

contracture
shoulder
paralysis obstetric


Neural oscillations in antipsychotic-naïve patients with a first psychotic episode.

Abstract:
OBJECTIVES
In chronic schizophrenic psychoses, oscillatory abnormalities predominantly occur in prefrontal cortical regions and are associated with reduced communication across cortical areas. Nevertheless, it remains unclear whether similar alterations can be observed in patients with a first episode of psychosis (FEP), a state characterised by pathological features occurring in both late prodromal patients and initial phases of frank schizophrenic psychoses.

METHODS
We assessed resting-state electroencephalographic data of 31 antipsychotic-naïve FEP patients and 29 healthy controls (HC). We investigated the three-dimensional (3D) current source density (CSD) distribution and lagged phase synchronisation (LPS) of oscillations across small-scale and large-scale brain networks. We additionally investigated LPS relationships with clinical symptoms using linear mixed-effects models.

RESULTS
Compared to HC, FEP patients demonstrated abnormal CSD distributions in frontal areas of the brain; while decreased oscillations were found in the low frequencies, an increase was reported in the high frequencies (P < 0.01). Patients also exhibited deviant LPS in the high frequencies, whose dynamics changed over increasing 3D cortico-cortical distances and increasing psychotic symptoms.

CONCLUSIONS
These results indicate that in addition to prefrontal cortical abnormalities, altered synchronised neural oscillations are also present, suggesting possible disruptions in cortico-cortical communications. These findings provide new insights into the pathophysiological mechanisms of emerging schizophrenic psychoses.

Ramyead|Avinash|A| Studerus|Erich|E| Kometer|Michael|M| Heitz|Ulrike|U| Gschwandtner|Ute|U| Fuhr|Peter|P| Riecher-Rössler|Anita|A|

psychosis
EEG
Schizophrenia
biomarkers
phase synchrony


Reconstruction of oral cavity defect using versatile buccinator myomucosal flaps in the treatment of cT2-3, N0 oral cavity squamous cell carcinoma: Feasibility, morbidity, and functional/oncological outcomes.

Abstract:
OBJECTIVES
To evaluate the results of oral cavity reconstruction using various types of buccinator myomucosal flap (BMMF) for the treatment of cT2-3 oral cavity squamous cell carcinoma (OCSCC).

MATERIALS AND METHODS
Twenty-two consecutive patients who underwent surgical management for OCSCC and reconstruction using various types of BMMF were enrolled. Surgical results, complications, speech/swallowing outcomes, and recurrence were evaluated.

RESULTS
The mean flap harvest time was 28.6 min. Complete flap necrosis occurred in one (4.5%) patient in whom an island flap was used. There were no other major complications. All patients demonstrated normal speech and swallowing functions, with articulation correctness >90% and a mean swallowing satisfaction score of 9.6. Occult neck lymph node metastasis was found in three (13.6%) patients; however, no recurrence at level I was observed.

CONCLUSIONS
Reconstruction of the oral cavity using various types of BMMF is feasible for treatment of cT2-3 OCSCC, and yields favorable outcomes with minimal morbidity.

Ahn|Dongbin|D|
Lee|Gil Joon|GJ|
Sohn|Jin Ho|JH|

Reconstruction
Flap
Buccal mucosa
Cheek
Oral cancer


Mapping Arginase Expression with 18F-Fluorinated Late-Generation Arginase Inhibitors Derived from Quaternary α-Amino Acids.

Abstract: Arginase hydrolyzes L-arginine and influences levels of polyamines and nitric oxide. Arginase overexpression is associated with inflammation and tumorigenesis. Thus, radiolabeled arginase inhibitors may be suitable PET tracers for staging arginase-related pathophysiologies. We report the synthesis and evaluation of 2 radiolabeled arginase inhibitors, 18F-FMARS and 18F-FBMARS, developed from α-substituted-2-amino-6-boronohexanoic acid derivatives. Methods: Arylboronic ester-derived precursors were radiolabeled via copper-mediated fluorodeboronation. Binding assays using arginase-expressing PC3 and LNCaP cells were performed. Autoradiography of lung sections from a guinea pig model of asthma overexpressing arginase and dynamic small-animal PET imaging with PC3-xenografted mice evaluated the radiotracers' specific binding and pharmacokinetics. Results: 18F-fluorinated compounds were obtained with radiochemical yields of up to 5% (decay-corrected) and an average molar activity of 53 GBq⋅μmol-1 Cell and lung section experiments indicated specific binding that was blocked up to 75% after pretreatment with arginase inhibitors. Small-animal PET studies indicated fast clearance of the radiotracers (7.3 ± 0.6 min), arginase-mediated uptake, and a selective tumor accumulation (SUV, 3.0 ± 0.7). Conclusion: The new 18F-fluorinated arginase inhibitors have the potential to map increased arginase expression related to inflammatory and tumorigenic processes. 18F-FBMARS showed the highest arginase-mediated uptake in PET imaging and a significant difference between uptake in control and arginase-inhibited PC3 xenografted mice. These results encourage further research to examine the suitability of 18F-FBMARS for selecting patients for treatments with arginase inhibitors.

Clemente|Gonçalo S|GS|
Antunes|Inês F|IF|
Kurhade|Santosh|S|
van den Berg|Mariska P M|MPM|
Sijbesma|Jürgen W A|JWA|
van Waarde|Aren|A|
Buijsman|Rogier C|RC|
Willemsen-Seegers|Nicole|N|
Gosens|Reinoud|R|
Meurs|Herman|H|
Dömling|Alexander|A|
Elsinga|Philip H|PH|


POLG-Associated Ataxia Presenting as a Fragile X Tremor/Ataxia Phenocopy Syndrome.

Abstract: Hyperintensities in the middle cerebellar peduncles (MCP), known as the MCP sign, and progressive late-onset ataxia constitute major characteristics of the fragile X tremor/ataxia syndrome (FXTAS). Here, we describe a 60-year-old male affected by ataxia due to biallelic mutations in the mitochondrial polymerase gamma (POLG) gene in which hyperintensities of the middle cerebellar peduncles (MCP) were found. The initial suspicion of FXTAS was however ruled out by a normal CGG expansion size in the FMR1 gene. We discuss the features of late-onset POLG-A as a phenocopy of FXTAS.

Paucar|Martin|M| Engvall|Martin|M| Gordon|Lisa|L| Tham|Emma|E| Synofzik|Matthis|M| Svenningsson|Per|P|

D051860:Fragile X Mental Retardation Protein


Higher PEDOT Molecular Weight Giving Rise to Higher Thermoelectric Property of PEDOT:PSS: A Comparative Study of Clevios P and Clevios PH1000.

Abstract: Poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) is a promising candidate as the next-generation thermoelectric (TE) material. Its TE properties are strongly dependent on its chemical and electronic structures. In this paper, we investigated the effect of PEDOT molecular weight on the TE properties of PEDOT:PSS films by a comparative study on two commercial grades of PEDOT:PSS, Clevios P, and Clevios PH1000. Dynamic light scattering (DLS) and Raman spectra imply that the PEDOT of Clevios PH1000 possesses longer conjugated chains than that of Clevios P. The TE properties of both the Clevios P and Clevios PH1000 films can be significantly enhanced through various post treatments, including solvent treatment, germinal diol treatment, organic solution treatment, and acid treatment. After these treatments, the treated Clevios PH1000 films constantly show both superior Seebeck coefficients and electrical conductivities over the treated Clevios P films. It is attributed to the higher molecular weight of PEDOT for the former than the latter. For the treated Clevios PH1000, longer PEDOT chains result in large PEDOT domains, facilitating the charge conduction a semimetallic behavior. Tuning the oxidation level of PEDOT:PSS is a facile way to enhance their TE property. A base treatment with sodium hydroxide was subsequently performed on both the treated Clevios P and Clevios PH1000 films. The power factors of both grades of PEDOT:PSS films were remarkably increased by a factor of 1.2-3.6. Still, both the conductivity and the Seebeck coefficient of a based-treated Clevios PH1000 film are superior over those of a control Clevios P film. The highest power factor the former is 334 μW/(m K2) for the former while only 11.4 μW/(m K2) for the latter. They are different by a factor of about 30 times.

molecular weight
thermoelectric property
PEDOT:PSS
post treatment
Seebeck coefficient
electrical conductivity


Medium to long-term results of a recessed glenoid for glenoid resurfacing in total shoulder arthroplasty.

Abstract:
Background
Recessed mini-glenoid components provide an alternative to total shoulder replacement that may avoid some of the known shortcomings and complications associated with shoulder hemiarthroplasty or standard glenoid components in difficult cases. This study reports survivorship, radiological and clinical outcomes of a recessed mini-glenoid implant in a consecutive cohort.

Methods
Retrospective cohort study reporting outcomes of 28 consecutive shoulders (27 patients) following total shoulder replacement using a recessed, cemented mini-glenoid implant at two sites.

Results
The most frequent diagnosis was primary osteoarthritis (79%); glenoid morphology was Walch Type A (67%), B1 15%, B2 10% and C 10%. At final follow-up, pain was 16.3 (SD = 23.1), American Shoulder and Elbow Score was 64.5 (SD = 31.9) and (normalized) Constant score was 83.0 (SD = 20.7). Implant survivorship at average final follow-up of seven years (3-13) was 96.4%. Seven mini-glenoids showed small peripheral radiolucent lines at one-year X-ray follow-up but were non-progressive on subsequent imaging.

Discussion
Recessed polyethylene mini-glenoid is an attractive alternative for shoulder arthroplasty and provides an intermediate solution between standard glenoid components and hemiarthroplasty. Our medium to long-term results demonstrate reliable clinical outcomes, absence of glenoid erosion, low complication rate and satisfactory implant survivorship.

recessed
glenoid
shoulder arthroplasty
shoulder replacement
osteoarthritis
custom implant


Characterization of influenza H1N1 Gag virus-like particles and extracellular vesicles co-produced in HEK-293SF.

Abstract: One of the concerns associated with the use of influenza virus-like particles (VLPs) as vaccine candidate or delivery system is their heterogeneous composition. Enveloped VLPs take up the host cell membrane at the budding site carrying out not only the viral antigenic proteins but also host proteins. In addition, the intrinsic nature of cells to produce membrane derived vesicles or extracellular vesicles (EVs), which have similar size to the VLPs, makes VLP purification process challenging. To further characterize these particles and identify proteins that are unique to each population, comparative proteomic analyses were completed to ultimately provide guidance for rational design of separation protocols. The VLPs were produced in suspension and serum free media by transient transfection of an inducible clone of a Human Embryonic Kidney (HEK-293SF) cells expressing HA and NA (H1N1/A/Puerto Rico/8/34), with a plasmid containing the gag gene of HIV-1 fused to GFP. EVs were produced independently from the non-transformed HEK-293SF cell line as a control for comparative studies. Both preparations were characterized for total nucleic acids and protein concentrations and extensively analyzed by nanoLC-MS/MS for their protein compositions. The proteomic analyses showed that aside from the recombinant VLP proteins, nucleolin was the most abundant host cell protein uniquely identified within VLPs (considering the MASCOT score value) while lactotransferrin and heat shock protein 90 were the most abundant proteins in EVs. Overall, this comparative study identifies potential target proteins as specific markers to guide VLP purification and discusses the biogenesis of enveloped particles released in HEK-293 cell suspension cultures emphasizing on the biological functions of host cell proteins identified.

Venereo-Sánchez|Alina|A| Fulton|Kelly|K| Koczka|Krisztina|K| Twine|Susan|S| Chahal|Parminder|P| Ansorge|Sven|S| Gilbert|Rénald|R| Henry|Olivier|O| Kamen|Amine|A|

D054301:gag Gene Products, Human Immunodeficiency Virus

Virus-like particles
Extracellular vesicles
HEK-293SF
Exosomes
Influenza vaccine
nLC-MS/MS


Quantitative analysis of noncoding RNA from paired fresh and formalin-fixed paraffin-embedded brain tissues.

Abstract: Formalin-fixed paraffin-embedded (FFPE) tissues are commonly used both clinically and in forensic pathology. Recently, noncoding RNA (ncRNA) has attracted interest among molecular medical researchers. However, it remains unclear whether newly identified ncRNAs, such as long noncoding RNA (lncRNA) and circular RNA (circRNA), remain stable for downstream molecular analysis in FFPE tissues. Here, we assessed the feasibility of using autoptic FFPE brain tissues from eight individuals to perform quantitative molecular analyses. Selected RNA targets (9 mRNAs and 15 ncRNAs) with different amplicon lengths were studied by RT-qPCR in paired fresh and FFPE specimens. For RNA quality assessment, RNA purity and yield were comparable between the two sample cohorts; however, the RNA integrity number decreased significantly during FFPE sampling. Amplification efficiency also displayed certain variability related with amplicon length and RNA species. We found molecular evidence that short amplicons of mRNA, lncRNA, and circRNA were amplified more efficiently than long amplicons. With the assistance of RefFinder, 5S, SNORD48, miR-103a, and miR-125b were selected as reference genes given their high stability. After normalization, we found that short amplicon markers (e.g., ACTB mRNA and MALAT1 lncRNA) exhibited high consistency of quantification in paired fresh/FFPE samples. In particular, circRNAs (XPO1, HIPK3, and TMEM56) presented relatively consistent and stable expression profiles in FFPE tissues compared with their corresponding linear transcripts. Additionally, we evaluated the influence of prolonged storage time on the amplification of gene transcripts and found that short amplicons still work effectively in archived FFPE biospecimens. In conclusion, our findings demonstrate the possibility of performing accurate quantitative analysis of ncRNAs using short amplicons and standardized RT-qPCR assays in autopsy-derived FFPE samples.

D021141:Nucleic Acid Amplification Techniques

D020133:Reverse Transcriptase Polymerase Chain Reaction

Real-time quantitative polymerase chain reaction (RT-qPCR)


Expression, purification, crystallization and preliminary crystallographic analysis of human myotubularin-related protein 3.

Abstract: Myotubularin-related proteins are a large family of phosphatases that have the catalytic activity of dephosphorylating the phospholipid molecules phosphatidylinositol 3-phosphate and phosphatidylinositol 3,5-bisphosphate. Each of the 14 family members contains a phosphatase catalytic domain, which is inactive in six family members owing to amino-acid changes in a key motif for the activity. All of the members also bear PH-GRAM domains, which have low homologies between them and have roles that are not yet clear. Here, the cloning, expression, purification and crystallization of human myotubularin-related protein 3 encompassing the PH-GRAM and the phosphatase catalytic domain are reported. Preliminary X-ray crystallographic analysis shows that the crystals diffracted to 3.30 Å resolution at a synchrotron X-ray source. The crystals belonged to space group C2, with unit-cell parameters a = 323.3, b = 263.3, c = 149.4 Å, β = 109.7°.

Son|Ji Young|JY|
Lee|Jee Un|JU|
Yoo|Ki-Young|KY|
Shin|Woori|W|
Im|Dong-Won|DW|
Kim|Seung Jun|SJ|
Ryu|Seong Eon|SE|
Heo|Yong-Seok|YS|

D054558:Protein Tyrosine Phosphatases, Non-Receptor

myotubularin-related proteins
phosphatase
PH-GRAM domain
MTMR3
phosphoinositide


Bidirectional selective genotyping approach for the identification of quantitative trait loci controlling earliness per se in winter rye (Secale cereale L.).

Abstract: The genes controlling earliness of plants include genes responsible for vernalisation (Vrn) and photoperiod (Ppd), and those that are not entirely associated with a response to temperature or light. The last group of loci is known as earliness per se (Eps). Eps genes have been most commonly reported in the scientific literature as quantitative trait loci (QTL). The objective of this study was to use a bidirectional selective genotyping (BSG) method for the identification of loci controlling Eps, assessed at the heading stage in a newly developed rye population of recombinant inbred lines. We identified four linkage groups representing fragments of chromosomes, 1R, 5R, 6R and 7R, and containing, in total, 155 DArT markers significantly associated with earliness in rye. The results are discussed in the context of previously published QTL analyses.

D002874:Chromosome Mapping
D032461:Chromosomes, Plant
D018744:DNA, Plant
D008040:Genetic Linkage
D005819:Genetic Markers
D060005:Genotyping Techniques
D040641:Quantitative Trait Loci
D012434:Secale