bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigation of p16INK4a as a prognostic biomarker in oral epithelial dysplasia
Authors: Nankivell Paul , Williams Hazel , Webster Keith , Pearson David , High Alec , MacLennan Kenneth , Senguven Burcu , McConkey Christopher , Rabbitts Pamela , Mehanna Hisham
Published: December 2013

Background

Human papilloma virus is a risk factor for oropharyngeal cancer. Evidence for a similar aetiological role in the development of oral dysplasia or its transformation to oral cancer is not as clear. Meta‐analyses estimate the prevalence of high‐risk human papilloma virus (HPV) serotypes to be three times higher in pre‐malignant lesions and cancer than in normal oral mucosa. However, this does not imply a causal relationship. Conflicting results are reported from the few studies examining the prognostic significance of HPV positivity in the development of oral cancer.

We aimed to examine the ability of p16INK4a protein expression, a surrogate marker of HPV infection, to predict malignant progression in a large cohort of oral dysplasia patients.

Methods

One hundred forty eight oral dysplasia cases underwent immunohistochemical analysis using a monoclonal antibody against p16INK4a. Clinical factors were also collated on each case. Slides were double scored independently by two trained observers. Univariate analyses using both logistic and Cox regression models were performed.

Results

Thirty nine of 148 cases progressed to cancer. Ten of 148 cases (7%) were p16INK4a positive. High grade of dysplasia (P = 0.0002) and lesion morphology (P = 0.03) were found to be prognostic of malignant progression. p16INK4a score was not prognostic in this cohort (P = 0.29). This did not change with a time to event analysis (P = 0.24).

Conclusion

Few studies have assessed the aetiological role of HPV in cancer development from dysplastic lesions. Our study, using one of the largest cohorts of oral dysplasia, demonstrated a low rate of p16INK4a positivity and was unable to confirm a prognostic ability for this biomarker.
Fibroblast growth factor 2 involved in the pathogenesis of synovial chondromatosis of temporomandibular joint
Authors: Li Yingjie , Cai Hengxing , Fang Wei , Meng Qinggong , Li Jian , Deng Mohong , Long Xing
Published: December 2013

Background

Synovial chondromatosis (SC) of temporomandibular joint (TMJ) is a rare proliferative disorder characterized by the formation of cartilaginous or osteocartilaginous nodules in synovium and joint space. Fibroblast growth factor 2 (FGF‐2) is frequently applied in chondrogenic differentiation assays. Therefore, we hypothesized that FGF‐2 might involved in the pathogenesis of SC.

Methods

SC synovium and loose bodies (LBs) specimens were observed by histological and immunohistochemical methods. Real‐time PCR was conducted for comparing genes expressions in SC and normal synovium. SC synoviocytes were stimulated by FGF‐2 in the presence or absence of its antagonist long pentraxin‐3 (PTX3) for 6 days. Real‐time PCR and alkaline phosphatase (ALP) activity were performed to examine the effects exerted by FGF‐2 and PTX3.

Results

SC synovium, no matter facing the articular cavity or covering LB, was characterized by increased quantity of synoviocytes and blood vessels. FGF‐2 was expressed in chondrocytes and fibroblast‐like cells of LBs, and the wall of blood vessels. Expressions of chondrogenic genes (Sox9 and Wnt‐4), osteogenic genes (Foxc2), FGF‐2, and VEGF‐A mRNA were significantly higher in SC synovium than that of the control group. The stimulation of FGF‐2 on SC synoviocytes increased ALP activity and expressions of chondrogenic genes (Sox9, Col2α1, and Aggrecan), osteogenic genes (Foxc2, osteocalcin, and Col1α1), and VEGF‐A, but PTX3 inhibited these effects.

Conclusion

FGF‐2 was responsible for the formation of cartilaginous loose bodies and involved in the pathogenesis of SC.
The Gudaga Study: Development in 3‐year‐old urban Aboriginal children
Authors: McDonald Jenny , Webster Vana , Knight Jennifer , Comino Elizabeth
Published: December 2013

Aim

The aim of this analysis was to study and explore factors associated with the developmental progress in urban Aboriginal children at 3 years.

Methods

The Gudaga Study is a longitudinal birth cohort study of urban Aboriginal infants. The children were assessed using the Griffiths Mental Development Scales, Extended Revised (GMDS‐ER) and the Peabody Picture Vocabulary Test, Fourth Edition (PPVT‐IV). Student's t‐tests and multiple linear regression analysis were used to test the association between developmental progress and possible risk factors.

Results

Overall, the mean general quotient (GQ) for Gudaga children was significantly lower than the standardised norm (P < 0.001). In the GMDS‐ER subscales, the scores were higher than expected in the locomotor (P = 0.002) and personal–social domains (P = 0.002) and lower than expected for language (P < 0.001), eye and hand coordination (P < 0.001), performance (P < 0.001) and practical reasoning (P < 0.001). Multiple regression analysis showed that maternal age (P = 0.02) and single‐mother status (P = 0.04) were significantly associated with lower performance on the GMDS‐ER. The GQ was inversely proportional to the number of risk factors present (P = 0.001). The mean score of the PPVT‐IV was also lower than the PPVT‐IV norms (P < 0.001).

Conclusion

At 3 years, urban Aboriginal children show relative strengths in their locomotor and self‐care skills and emerging delays in their language, fine motor and performance skills. Slower developmental progress was more likely in the context of young maternal age and single parenthood.
A Technique Combining Immunoprecipitation and RT‐PCR for RNA Plant Virus Detection
Authors: Lima José Albersio Araujo , Nascimento Aline Kelly Queiroz , Radaelli Paula , Silva Ana Kelly Firmino , Silva Fabiana Rodrigues
Published: December 2013

Plant virus identification and characterization can be accomplished by several methods involving their morphological, physical, biological, cytological, serological and molecular properties. The use of molecular techniques is increasing worldwide, and some have been developed for identification and characterization of plant viruses. Reverse transcription polymerase chain reaction (RT‐PCR) has been shown to be a suitable method for research with RNA plant viruses. In this study, a new approach of RT‐PCR involving previous virus immunoprecipitation (IP) was used for RNA amplification of five virus species of the genera Comovirus, Cucumovirus, Potyvirus and Sobemovirus from infected plant tissues. IP‐RT‐PCR was practical, sensitive and minimized problems with total RNA extractions from infected tissues. The technique provides partial virus particle purification by its specific immunoprecipitation, and it should be especially useful for RNA amplification of viruses that occur in low or variable concentrations in plant tissues or when the tissues contain various forms of RT‐PCR amplification inhibitors.
Occurrence of Meloidogyne spp. in Cerrado Vegetations and Reaction of Native Plants to Meloidogyne javanica
Authors: Silva Joelma G. P. , Furlanetto Cleber , Almeida Maria R. A. , Rocha David B. , Mattos Vanessa S. , Correa Valdir R. , Carneiro Regina M. D. G.
Published: December 2013

The Cerrado biome represents a hotspot of biodiversity. Despite this, the nematofauna in this biome has not been well characterized, especially that related to root‐knot nematodes. This work aimed to identify Meloidogyne species present in different cerrado vegetations and to investigate potential hosts of Meloidogyne javanica in this biome. Soil samples (250) were collected in native areas of cerrado vegetation located at the National Park of Brasília (PNB) (125 samples) and Água Limpa Farm (FAL) (125 samples), and transferred to sterile pots. Single tomato plants cv. Santa Clara (susceptible) were transplanted into individual pots and maintained for 90 days under glasshouse. Females of Meloidogyne spp. were extracted from tomato roots and identified based upon esterase phenotypes and confirmed with PCR using specific sequence characterized amplified regions (SCAR) primers. Native plants were inoculated with 10 000 individuals (eggs + J2) of a pure culture of M. javanica and maintained under glasshouse for 6 months. From the 250 samples collected, 57 (22.8%) presented Meloidogyne spp. A total of 66 Meloidogyne populations were identified as follows: M. javanica (75.76%), M. incognita (10.60%), M. hapla (9.1%), M. morocciensis (3.03%) and M. arenaria (1.51%). The following esterase phenotypes were detected: M. javanica (J3 and J2), M. incognita (I1 and I2), M. hapla (H1), M. morocciensis (A3) and M. arenaria (A2). The SCAR primers incK14F/incK14R, Fjav/Rjav and Fh/Rh amplified specific fragments in M. incognita (399 bp), M. javanica (670 bp) and M. hapla (610 bp) and can be used for identification of indigenous Meloidogyne spp. from cerrado. The primer set Far/Rar is not specific for M. arenaria due to the amplification of DNA in M. morocciensis. Mimosa caesalpiniifolia was the only native plant in which M. javanica developed a high reproductive rate, and it is probably a host for this nematode in cerrado.
Eugenol derivatives as potential anti-oxidants: is phenolic hydroxyl necessary to obtain an effect?
Authors: d' Avila Farias Marília , Oliveira Pathise Souto , Dutra Filipe S Pereira , Fernandes Thiely Jacobsen , de Pereira Claudio M P , de Oliveira Simone Quintana , Stefanello Francieli Moro , Lencina Claiton Leonetti , Barschak Alethéa Gatto
Published: December 2013

Objectives

Eugenol, obtained from clove oil (Eugenia caryophyllata), possess several biological activities. It is anti-inflammatory, analgesic, anaesthesic, antipyretic, antiplatelet, anti-anaphylactic, anticonvulsant, anti-oxidant, antibacterial, antidepressant, antifungal and antiviral. The anti-oxidant activity of eugenol have already been proven. From this perspective testing, a series of planned structural derivatives of eugenol were screened to perform structural optimization and consequent increase of the potency of these biological activities.

Methods

In an attempt to increase structural variability, 16 compounds were synthesized by acylation and alkylation of the phenolic hydroxyl group. Anti-oxidant activity capacity was based on the capture of DPPH radical (2,2-diphenyl-1-picryl-hydrazyl), ABTS radical 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), measure of TBARS (thiobarbituric acid-reactive species), total sulfhydryl and carbonyl content (eugenol derivatives final concentrations range from 50 to 200 μm).

Key findings

Four derivatives presented an efficient concentration to decrease 50% of the DPPH radical (EC50) < 100 μm, which has a good potential as a free-radical scavenger. Three of these compounds also showed reduction of ABTS radical. Eugenol derivatives presenting alkyl or aryl (alkylic or arylic) groups substituting hydroxyl 1 of eugenol were effective in reducing lipid peroxidation, protein oxidative damage by carbonyl formation and increase total thiol content in cerebral cortex homogenates. In liver, the eugenol derivatives evaluated had no effect.

Conclusions

Our results suggest that these molecules are promising anti-oxidants agents.
Molecular deficiency (ies) in MT1 melatonin signaling pathway underlies the melatonin‐unresponsive phenotype in MDA‐MB‐231 human breast cancer cells
Authors: Mao Lulu , Yuan Lin , Xiang Shulin , Zeringue Samantha B. , Dauchy Robert T. , Blask David E. , Hauch Adam , Hill Steven M.
Published: December 2013

Melatonin has been shown repeatedly to inhibit the growth of human breast tumor cells in vitro and in vivo. Its antiproliferative effects have been well studied in MCF‐7 human breast cancer cells and several other estrogen receptor α (ERα)‐positive human breast cancer cell lines. However, the MDA‐MB‐231 breast cancer cell line, an ERα‐negative cell line widely used in breast cancer research, has been shown to be unresponsive to melatonin's growth‐suppressive effect in vitro. Here, we examined the effect of melatonin on the cell proliferation of several ERα‐negative breast cancer cell lines including MDA‐MB‐231, BT‐20, and SK‐BR‐3 cells. Although the MT1 G‐protein‐coupled receptor is expressed in all three cell lines, melatonin significantly suppressed the proliferation of SK‐BR‐3 cells without having any significant effect on the growth of MDA‐MB‐231 and BT‐20 cells. We confirmed that the MT1‐associated Gα proteins are expressed in MDA‐MB‐231 cells. Further studies demonstrated that the melatonin unresponsiveness in MDA‐MB‐231 cells may be caused by aberrant signaling downstream of the Gαi proteins, resulting in differential regulation of ERK1/2 activity.
2‐Amino‐N‐(2,6‐dichloropyridin‐3‐yl)acetamide derivatives as a novel class of HBV capsid assembly inhibitor
Authors: Cho M. H. , Jeong H. , Kim Y. S. , Kim J.‐W. , Jung G.
Published: December 2013

Capsid structure is crucial for the maturation and maintenance of the stable hepatitis B virion. Therefore, chemicals that inhibit capsid assembly might potentially act as potent antiviral compounds. However, only a few chemicals are known to block the capsid assembly process and further viral proliferation. In this study, we present a novel family of capsid assembly inhibitors that act against hepatitis B virus (HBV). Based on X‐ray crystallographic data of the HBV core protein (Cp), we built dimer and hexamer structural models to be used in library searches. Several chemicals in the 2‐amino‐N‐(2,6‐dichloropyridin‐3‐yl)acetamide family were predicted to have high affinity for the groove structure in Cp. Using in vitro assembly and the HepG2.2.15 cell culture test, we verified that these chemicals demonstrated inhibitory effects on capsid assembly. Furthermore, we investigated the combinatorial effects of these assembly inhibitor chemicals with lamivudine and revealed that, in combination, they have synergistic inhibitory effects on decreasing viral concentration. We propose that these inhibitors could be utilized as an effective combination treatment against HBV infection.
Treatment of Aldosterone‐Secreting Adrenocortical Tumors in Cats by Unilateral Adrenalectomy: 10 Cases (2002–2012)
Authors: Lo A.J. , Holt D.E. , Brown D.C. , Schlicksup M.D. , Orsher R.J. , Agnello K.A.
Published: December 2013

Background

Primary hyperaldosteronism (PHA) in cats occurs as a consequence of excessive hormone production by an adrenocortical tumor. Median survival time, association between tumor type and prognosis, and the likelihood that cats require continued medical therapy after surgery have not been systematically evaluated.

Objectives

To determine the median survival time of cats with PHA treated by unilateral adrenalectomy. To examine if tumor type, anesthesia time, or tumor location (left or right side) affect survival and if affected cats require continued postoperative treatment for persistent hypertension or hypokalemia.

Animals

Ten client‐owned cats.

Methods

Retrospective study. Cats were diagnosed with PHA based on clinical signs, increased plasma aldosterone concentration, and advanced imaging. Cats underwent unilateral adrenalectomy. Survival time (days alive after surgery) was determined for each cat. Factors affecting median survival time were investigated, including histopathology, anesthesia time, and location (side) of the tumor.

Results

Eight of 10 cats survived to discharge from the hospital post adrenalectomy. Overall median survival was 1,297 days (range 2–1,582 days). The only significant factor affecting median survival time was anesthesia time >4 hours. Tumor type and location (side) did not significantly affect median survival time. No cats required continued medical treatment for PHA.

Conclusions and Clinical Importance

Although PHA in cats is still considered an uncommon condition, it should be considered in middle to older aged cats with hypokalemic polymyopathy and systemic hypertension. Surgical correction by unilateral adrenalectomy is a viable approach to definitive treatment of PHA with no need for continued medical management.
Evaluation of the Effects of a Therapeutic Renal Diet to Control Proteinuria in Proteinuric Non‐Azotemic Dogs Treated with Benazepril
Authors: Cortadellas O. , Talavera J. , Fernández del Palacio M.J.
Published: December 2013

Background

Angiotensin‐converting enzyme inhibitors (ACEIs) are currently used to control proteinuria in dogs with chronic kidney disease. Renal diets (RDs) have beneficial effects in the management of azotemic dogs, but its role in proteinuric non‐azotemic (PNAz) dogs has been poorly documented.

Hypothesis

Administration of a RD to PNAz dogs treated with benazepril (Be) improves proteinuria control compared with the administration of a maintenance diet (MD).

Animals

Twenty‐two PNAz (urine protein/creatinine ratio [UPC] >1) dogs.

Methods

Randomized open label clinical trial design. Dogs were assigned to group‐MD (5.5 g protein/100 kcal ME)/Be or to group‐RD (3.7 g protein/100 kcal ME)/Be group during 60 days. Dogs with serum albumin (Alb) <2 g/dL received aspirin (1 mg/kg/12 hours). A physical examination, systolic blood pressure (SBP) measurement, complete blood count (CBC), biochemistry panel, urinalysis, and UPC were performed at day 0 (D0) and day 60 (D60).

Results

At D0, there were no significant differences between groups in the evaluated variables. During the study, logUPC (geometric mean (95% CI) and SBP (mean±SD mmHg) significantly decreased (paired t‐test, P = 0.001) in Group‐RD (logUPCD0 = 3.16[1.9–5.25]; UPCD60 = 1.20 [0.59–2.45]; SBPD0 = 160 ± 17.2; SBPD60 = 151 ± 15.8), but not in Group‐MD (UPCD0 = 3.63[2.69–4.9]; UPCD60 = 2.14 [0.76–6.17]; SBPD0 = 158 ± 14.7; SBPD60 = 153 ± 11.5). However, RM‐ANOVA test did not confirm that changes were consequence of dietary modification. Weight and Alb concentration did not change significantly in any group.

Conclusion and Clinical Relevance

The administration of a RD to PNAz dogs treated with Be might help to control proteinuria and SBP compared with the administration of a MD, without inducing clinically detectable malnutrition, but more studies are warranted.
Serum Adipokine Concentrations in Dogs with Naturally Occurring Pituitary‐Dependent Hyperadrenocorticism
Authors: Cho K.‐D. , Paek J. , Kang J.‐H. , Chang D. , Na K.‐J. , Yang M.‐P.
Published: December 2013

Background

An excess of intra‐abdominal fat is observed frequently in dogs with hyperadrenocorticism (HAC). Adipokine dysregulation is a possible cause of complications related to visceral obesity, but little information is available on adipokine in dogs with naturally occurringHAC.

Objectives

To examine the differences in the circulating adipokines concentrations in overweight dogs with and without pituitary‐dependentHAC(PDH).

Animals

Thirty healthy dogs and 15 client‐owned dogs withPDH.

Methods

Case–controlled observational study, which enrolled 15 overweight dogs diagnosed withPDHand 30 otherwise healthy dogs of similar body condition score. Nine of 15 dogs withPDHwere treated with low‐dose trilostane twice daily and reassessed after treatment.

Results

The serum leptin (P < .0001) and insulin (P < .0001) concentrations were significantly higher in thePDHgroup (leptin, 22.8 ± 8.8 [mean ± SD]; insulin, 9.1 ± 6.1) than the healthy group (leptin, 4.9 ± 3.7; insulin, 1.9 ± 0.9). However, there were no significant differences in the adiponectin, resistin, tumor necrosis factor (TNF)‐α, interleukin (IL)‐1β,IL‐6,IL‐10, andIL‐18 levels between the 2 groups. In thePDHgroup, the serum cortisol concentrations had a linear association with the leptin concentrations, and there were significant decreases in the leptin (P = .0039) and insulin (P = .0039) levels after trilostane treatment. However, the leptin and insulin levels remained higher after trilostane treatment than in healthy control dogs with similar body condition score.

Conclusions and Clinical Importance

Hypercortisolemia in dogs withPDHmight upregulate the circulating leptin levels. However, a large population‐based study will be necessary to determine whether the upregulation of leptin is involved directly with the complications caused byHAC.
The influence of tree‐scale and ecosystem‐scale factors on epiphytic lichen communities across a long‐term retrogressive chronosequence
Authors: Asplund Johan , Sandling Aron , Kardol Paul , Wardle David A.
Published: December 2013

Questions

We tested the relationship between total cover, species richness and composition of epiphytic lichens on trunks of Betula pubescens and ecosystem retrogression (i.e. prolonged absence of major disturbance). We then investigated how the relationships changed when also accounting for tree‐scale factors (aspect, height and bark characteristics) and ecosystem‐scale factors (e.g. light transmission, tree species diversity and soil fertility).

Location

Thirty forested islands in northern Sweden differing in fire history, which collectively represent a retrogressive chronosequence spanning ca. 5000 yr.

Results

Total lichen cover responded negatively to long‐term absence of major disturbance, but only at exposed positions on the tree trunk, indicating that lichen cover on substrates with more favourable microclimates is less susceptible to environmental change at the ecosystem scale. Further, although there was no overall effect of island size on lichen species richness, we did find a significant interactive effect between island size and height on trunk on species richness. This emerged because species richness decreased with retrogression for lichen communities at breast height, but showed a hump‐shaped response to retrogression at the trunk base. Shifts in ecosystem properties with retrogression explained some of the variation in lichen community composition, but most of the variation could be explained by tree‐scale factors, notably height on the trunk.

Conclusions

While it has frequently been shown that lichens increase in abundance and richness during the first two or three centuries of succession, our results highlight that over a much longer time scale, encompassing soil aging and declining soil fertility, the lichen flora can be negatively affected. However, these effects are heavily mediated by tree‐scale factors. These changes in the lichen community may be of potential importance for ecosystem processes and higher trophic level interactions driven by lichen communities.
Guided reading: young pupils' perspectives on classroom practice
Authors: Hanke Veronica
Published: December 2013

Guided reading is widely perceived to be tricky in English primary schools; prior research has found difficulties with teacher interpretation and implementation. The study reported here suggests that to understand the problems associated with it we should also take into account pupils' perspectives on their guided reading lessons. In this case, the pupils were 4–7 years old. The special challenges of accessing young pupil perspectives were addressed through co‐authored drawings, a type of graphic elicitation. The drawings, together with other collected data, generated insights into pupil perceptions of literacy practices, of the role of friends and ability grouping in learning to read, and of their teachers' organisational challenges. A socio‐cultural analysis of these data indicates that these pupils were sensitive to the social and cultural contexts of their guided reading lessons. It may be that precise official advice for guided reading in England, originating in the National Literacy Strategy documents in 1999 and the Primary National Strategy documents in 2003 and 2008, has been insufficiently sensitive to the complex teaching and learning contexts of guided reading and that this discrepancy has contributed to the identified difficulties. The paper concludes that some rethinking of the literacy practice of guided reading would be valuable.
Sponge heterotrophic capacity and bacterial community structure in high‐ and low‐microbial abundance sponges
Authors: Poppell Ericka , Weisz Jeremy , Spicer Lori , Massaro Andrew , Hill April , Hill Malcolm
Published: December 2013

The ecological and evolutionary forces that shape interactions between marine sponges and their symbiotic microbiota remain poorly understood. Considerable work has been done to characterize the sponge microbiome, and this research has shown that there are two distinct types of sponges: those with high microbial abundances (HMA) and those with low microbial abundances (LMA). There is no clear evolutionary driver for this distinction, and we have only a limited understanding of how HMA or LMA status affects host phenotypes (e.g. feeding behavior, capacity for nutrient cycling). We had two primary goals with the present study. First, we used a variety of microscopic techniques to compare aspects of host biology (e.g. choanocyte chamber morphology and density) in the context of symbiont status. Secondly, we used molecular approaches to uncover components of ecological structure of bacterial communities in HMA versus LMA sponges (e.g. species richness, evenness). We found that choanocyte chamber density is greater in LMA sponges than in HMA sponges. We also found distinct patterns of organization for bacterial communities in HMA and LMA sponges, although environmental factors, to a lesser degree, also influence community structure in the Floridian sponges we examined. These results suggest that the large bacterial communities found in HMA sponges may allow the host to decrease their heterotrophy versus that of LMA sponges with minimal bacterial communities.
Reactive Oxygen Species‐Caspase‐3 Relationship in Mediating Blood–Brain Barrier Endothelial Cell Hyperpermeability Following Oxygen–Glucose Deprivation and Reoxygenation
Authors: Alluri Himakarnika , Stagg Hayden W. , Wilson Rickesha L. , Clayton Robert P. , Sawant Devendra A. , Koneru Madhavi , Beeram Madhava R. , Davis Matthew L. , Tharakan Binu
Published: December 2013

Objective

Microvascular hyperpermeability that occurs due to breakdown of the BBB is a major contributor of brain vasogenic edema, following IR injury. In microvascular endothelial cells, increased ROS formation leads to caspase‐3 activation following IR injury. The specific mechanisms, by which ROS mediates microvascular hyperpermeability following IR, are not clearly known. We utilized an OGD‐R in vitro model of IR injury to study this.

Methods

RBMEC were subjected to OGD‐R in presence of a caspase‐3 inhibitor Z‐DEVD, caspase‐3 siRNA or an ROS inhibitor L‐AA. Cytochrome c levels were measured by ELISA and caspase‐3 activity was measured fluorometrically. TJ integrity and cytoskeletal assembly were studied using ZO‐1 immunofluorescence and rhodamine phalloidin staining for f‐actin, respectively.

Results

OGD‐R significantly increased monolayer permeability, ROS formation, cytochrome c levels, and caspase‐3 activity (p < 0.05) and induced TJ disruption and actin stress fiber formation. Z‐DEVD, L‐AA and caspase‐3 siRNA significantly attenuated OGD‐R‐induced hyperpermeability (p < 0.05) while only L‐AA decreased cytochrome c levels. Z‐DEVD and L‐AA protected TJ integrity and actin cytoskeletal assembly.

Conclusions

These results suggest that OGD‐R‐induced hyperpermeability is ROS and caspase‐3 dependent and can be regulated by their inhibitors.
Inter‐viral conflicts that exploit host CRISPR immune systems of Sulfolobus
Authors: Erdmann Susanne , Le Moine Bauer Sven , Garrett Roger A.
Published: December 2013

Infection of Sulfolobus islandicus REY15A with mixtures of different Sulfolobus viruses, including STSV2, did not induce spacer acquisition by the host CRISPR immune system. However, coinfection with the tailed fusiform viruses SMV1 and STSV2 generated hyperactive spacer acquisition in both CRISPR loci, exclusively from STSV2, with the resultant loss of STSV2 but not SMV1. SMV1 was shown to activate adaptation while itself being resistant to CRISPR‐mediated adaptation and DNA interference. Exceptionally, a single clone S‐1 isolated from an SMV1 + STSV2‐infected culture, that carried STSV2‐specific spacers and had lost STSV2 but not SMV1, acquired spacers from SMV1. This effect was also reproducible on reinfecting wild‐type host cells with a variant SMV1 isolated from the S‐1 culture. The SMV1 variant lacked a virion protein ORF114 that was shown to bind DNA. This study also provided evidence for: (i) limits on the maximum sizes of CRISPR loci; (ii) spacer uptake strongly retarding growth of infected cultures; (iii) protospacer selection being essentially random and non‐directional, and (iv) the reversible uptake of spacers from STSV2 and SMV1. A hypothesis is presented to explain the interactive conflicts between SMV1 and the host CRISPR immune system.
Structural polymorphism in the promoter of pfmrp2 confers Plasmodium falciparum tolerance to quinoline drugs
Authors: Mok Sachel , Liong Kek‐Yee , Lim Eng‐How , Huang Ximei , Zhu Lei , Preiser Peter Rainer , Bozdech Zbynek
Published: December 2013

Drug resistance in Plasmodium falciparum remains a challenge for the malaria eradication programmes around the world. With the emergence of artemisinin resistance, the efficacy of the partner drugs in the artemisinin combination therapies (ACT) that include quinoline‐based drugs is becoming critical. So far only few resistance markers have been identified from which only two transmembrane transporters namely PfMDR1 (an ATP‐binding cassette transporter) and PfCRT (a drug‐metabolite transporter) have been experimentally verified. Another P. falciparum transporter, the ATP‐binding cassette containing multidrug resistance‐associated protein (PfMRP2) represents an additional possible factor of drug resistance in P. falciparum. In this study, we identified a parasite clone that is derived from the 3D7 P. falciparum strain and shows increased resistance to chloroquine, mefloquine and quinine through the trophozoite and schizont stages. We demonstrate that the resistance phenotype is caused by a 4.1 kb deletion in the 5′ upstream region of the pfmrp2 gene that leads to an alteration in the pfmrp2 transcription and thus increased level of PfMRP2 protein. These results also suggest the importance of putative promoter elements in regulation of gene expression during the P. falciparum intra‐erythrocytic developmental cycle and the potential of genetic polymorphisms within these regions to underlie drug resistance.
Detection and isolation of the α‐proteobacterium Asaia in Culex mosquitoes
Authors: DE FREECE C. , DAMIANI C. , VALZANO M. , D'AMELIO S. , CAPPELLI A. , RICCI I. , FAVIA G.
Published: December 2013

Investigations of microbiota within mosquitoes continue to widen the spectrum of possible symbiont‐based applications against vector‐borne diseases. In this context, α‐proteobacteria of the genus Asaia (Rhodospirillales: Acetobacteraceae) are emerging as possible endosymbiotic candidates, particularly in paratransgenic approaches aimed at interrupting pathogen transmission. Previous studies have shown that Asaia spp. distribution among Anopheles gambiae and Anopheles stephensi (Diptera: Culicidae) mosquitoes displayed positive rates of infection in isolated midguts, salivary glands and reproductive tissues. Similarly, Asaia has been detected in Aedes albopictus (Stegomyia albopicta) and Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) populations. Within the Culex pipiens complex (Diptera: Culicidae), Asaia infection is still largely unexplored. Here, we summarize a preliminary survey of laboratory‐reared Cx. pipiens complex and field‐collected Culex quinquefasciatus for the presence of Asaia spp., and present the first identification of Asaia in some of the members of the Cx. pipiens complex and the first description in West African populations of Cx. quinquefasciatus.
Metoclopramide nasal spray is effective in symptoms of gastroparesis in diabetics compared to conventional oral tablet
Authors: Parkman H. P. , Carlson M. R. , Gonyer D.
Published: December 2013

Background

Delayed gastric emptying symptoms interfere with the absorption of oral medications. Intranasal metoclopramide is being developed as an alternative to oral metoclopramide for patients with diabetic gastroparesis.

Methods

To compare the efficacy and safety of metoclopramide nasal spray to oral tablets in diabetic patients with symptoms of gastroparesis, this randomized, open‐label, parallel design study randomized subjects to 10 or 20 mg nasal spray or 10 mg tablet four times a day for 6 weeks. Efficacy was evaluated using a total symptom score (TSS).

Key Results

Eighty‐nine subjects were enrolled. For the intention to treat population, both nasal dose groups (10 and 20 mg) had lower TSS with treatment compared to the oral 10 mg group. The change from baseline in TSS for nasal 20 mg was greater than the oral 10 mg at Week 6 (p = 0.026). For the per‐protocol population, there was a significant difference in the TSS between baseline and Week 6 for both the nasal 10 mg (p = 0.026) and the nasal 20 mg (p = 0.008) cohorts compared to the oral 10 mg group. Based on the definition of a responder, 88.9% of subjects who received oral 10 mg, 91.2% who received nasal 10 mg, and 97.1% who received nasal 20 metoclopramide were classified as responders. The side‐effect profile of the metoclopramide nasal spray was favorable. More side effects, especially nausea, occurred with the oral tablets.

Conclusions & Inferences

Metoclopramide nasal spray offers better symptom control than metoclopramide oral tablet in diabetic patients with symptoms of gastroparesis.
Awakening is not a metaphor: the effects of Buddhist meditation practices on basic wakefulness
Authors: Britton Willoughby B. , Lindahl Jared R. , Cahn B. Rael , Davis Jake H. , Goldman Roberta E.
Published: December 2013

Buddhist meditation practices have become a topic of widespread interest in both science and medicine. Traditional Buddhist formulations describe meditation as a state of relaxed alertness that must guard against both excessive hyperarousal (restlessness) and excessive hypoarousal (drowsiness, sleep). Modern applications of meditation have emphasized the hypoarousing and relaxing effects without as much emphasis on the arousing or alertness‐promoting effects. In an attempt to counterbalance the plethora of data demonstrating the relaxing and hypoarousing effects of Buddhist meditation, this interdisciplinary review aims to provide evidence of meditation's arousing or wake‐promoting effects by drawing both from Buddhist textual sources and from scientific studies, including subjective, behavioral, and neuroimaging studies during wakefulness, meditation, and sleep. Factors that may influence whether meditation increases or decreases arousal are discussed, with particular emphasis on dose, expertise, and contemplative trajectory. The course of meditative progress suggests a nonlinear multiphasic trajectory, such that early phases that are more effortful may produce more fatigue and sleep propensity, while later stages produce greater wakefulness as a result of neuroplastic changes and more efficient processing.
Identification of vegetative heat‐tolerant upland cotton (Gossypium hirsutum L.) germplasm utilizing chlorophyll fluorescence measurement during heat stress
Authors: Wu Tingting , Weaver David B. , Locy Robert D. , McElroy Scott , van Santen Edzard
Published: December 2013

Heat stress adversely affects upland cotton (Gossypium hirsutum L.) production in the United States. Heat tolerance is difficult to determine, especially when yield and fibre quality are the parameters measured in photoperiodic, non‐adapted genotypes. Our objective was to evaluate wild (mostly photoperiodic, non‐adapted) genotypes for vegetative heat tolerance using chlorophyll fluorescence. Parameters were measured in three phases of heat stress assays to select candidate lines for introgression of increased heat tolerance into commercial cotton germplasm. Forty‐four wild accessions were selected from 1762 wild accessions in the USDA germplasm collection using a single‐leaf sample assay (phase I). In the growth chamber, 6‐week‐old cotton plants were subjected to heat stress at 45°C and 80% humidity for 24 h. After heat stress, the mean ΦPSII for the selected accessions was higher than that of a set of randomly chosen accessions and adapted cultivars (phase II). In the field under high temperatures (>35°C), the top nine accessions had higher average ΦPSII than adapted cultivars (phase III). Based on these results, the three‐phased stepwise approach was consistent in identifying putative genetic resources for increased vegetative heat tolerance. Accessions were identified that may have potential as genetic materials for development of adapted heat‐tolerant upland cotton germplasm.
Prevalence of depression symptoms and serum levels of interleukin‐6 in hemodialysis patients
Authors: Knuth Berenice , Radtke Vinícius , Rocha Pablo , da Silva Kátia Sulenir , Dalsóglio Fabiana , Gazal Marta , Jansen Karen , Souza Diogo Onofre , Portela Luis Valmor , Kaster Manuella , Oses Jean Pierre
Published: December 2013

Aim

In hemodialysis patients, depression appears as the most common psychopathological condition. States of advanced chronic kidney disease and dialysis are associated with a state of chronic inflammation. Depression has been linked to activation of the immune system characterized by high levels of pro‐inflammatory cytokines. In this study, we investigated the possible correlations between depression, and interleukin‐6 (IL‐6) in hemodialysis patients.

Methods

Seventy‐five hemodialysis patients were enrolled in a cross‐sectional study from September to November 2011 in Pelotas, Rio Grande do Sul. Demographic data were obtained from a questionnaire and the Beck Depression Inventory (BDI) was used to determine the presence or absence of depression symptoms. Biochemical parameters, dialysisdosage delivery, and IL‐6 serum levels were measured.

Results

Prevalence of depression among hemodialysis patients was 48% (BDI ≥ 14). In biochemical assessments, depressed patients showed a decrease in urea (P = 0.01) and increase of IL‐6 (P = 0.04) levels. The correlation analysis between BDI scores and the biochemical variables showed that BDI was negatively correlated with urea (P = 0.03) and potassium (P = 0.04), but not with IL‐6 levels.

Conclusion

Hemodialysis patients with depression showed higher levels of IL‐6 but the severity of depressive symptoms was not correlated with levels of this cytokine.
Sequence variation in the putative effector gene SIX8 facilitates molecular differentiation of Fusarium oxysporum f. sp. cubense
Authors: Fraser‐Smith S. , Czislowski E. , Meldrum R. A. , Zander M. , O'Neill W. , Balali G. R. , Aitken E. A. B.
Published: December 2013

Fusarium oxysporum f. sp. cubense (Foc), causal agent of fusarium wilt of banana, is among the most destructive pathogens of banana and plantain. The development of a molecular diagnostic capable of reliably distinguishing between the various races of the pathogen is of key importance to disease management. However, attempts to distinguish isolates using the standard molecular loci typically used for fungal phylogenetics have been complicated by a poor correlation between phylogeny and pathogenicity. Among the available alternative loci are several putative effector genes, known as SIX genes, which have been successfully used to differentiate the three races of F. oxysporum f. sp. lycopersici. In this study, an international collection of Foc isolates was screened for the presence of the putative effector SIX8. Using a PCR and sequencing approach, variation in Foc‐SIX8 was identified which allowed race 4 to be differentiated from race 1 and 2 isolates, and tropical and subtropical race 4 isolates to be distinguished from one another.
Predictors of mortality in rheumatoid arthritis‐related interstitial lung disease
Authors: Assayag Deborah , Lubin Molly , Lee Joyce S. , King Talmadge E. , Collard Harold R. , Ryerson Christopher J.
Published: December 2013

Rheumatoid arthritis‐associated interstitial lung disease (RA‐ILD) has a heterogeneous clinical presentation and disease course. Establishing prognosis for these patients is challenging. Identifying the factors that predict mortality in patients with RA‐ILD could help guide management. A detailed systematic review was conducted in order to identify individual variables that predict mortality in RA‐ILD. A literature review was performed using keywords and medical subject headings to identify all articles relating to the prognosis of RA‐ILD. Studies were included if they identified predictors of mortality in adults with RA‐ILD, were published in English, and included at least 10 patients with RA‐ILD. Two authors independently reviewed each citation and extracted data from all studies meeting inclusion criteria. Any differences were then resolved by consensus. A total of 10 studies met our inclusion criteria. All were observational cohort studies of variable quality. Mean age of reported patients ranged from 55 to 69 years, and 41.7% of all patients were male. Median survival ranged from 3.2 to 8.1 years. Significant predictors of mortality on multivariate analysis were older age, male gender, lower diffusion capacity for carbon monoxide, extent of fibrosis, and the presence of usual interstitial pneumonia pattern. Mortality in RA‐ILD is associated with several patient‐ and ILD‐specific variables; however, previous studies are of low quality.
Doping in sports: Knowledge and attitudes among parents of Austrian junior athletes
Authors: Blank C. , Leichtfried V. , Schaiter R. , Fürhapter C. , Müller D. , Schobersberger W.
Published: December 2013

Strategies for doping prevention are based on prior identification of opportunities for intervention. There is no current research focusing on the potential role in doping prevention, which might be played by the parents of junior elite athletes. The purpose of this study was to evaluate the knowledge and attitudes toward doping among parents of Austrian junior athletes and to analyze factors potentially influencing these beliefs. In this study, two questionnaires were distributed to 1818 student athletes, each with instructions that these surveys were to be completed by their parents (ntotal = 3636). Parents filled in questionnaires at home without observation. Responses from 883 parents were included in this analysis. Compared to female parents, male parents demonstrated significantly better knowledge about doping and its side effects and were more likely to be influenced by their own sporting careers and amounts of sports activities per week. Parental sex did not demonstrate a significant influence on responses reflecting attitudes toward doping. Additional research is needed to compare these results with young athletes' knowledge and attitudes to determine if and to what degree parental attitudes and beliefs influence the behavior and attitudes of their children.
Navigating the mtDNAroad map out of the morphological maze: interpreting morphological variation in the diverseMonomorium rothsteini(Forel) complex (Hymenoptera:Formicidae)
Authors: SPARKS KATHRYN S. , ANDERSEN ALAN N. , DONNELLAN STEPHEN C. , AUSTIN ANDREW D.
Published: December 2013

Monomoriumis a large and diverse ant genus with speciose radiations in both theAfrotropical andAustralian regions. According to the most recent taxonomic revision, manyAustralian species are characterised by very broad distributions and variable morphology, which suggests that some species may be unrecognised species complexes. With a continent‐wide distribution and diverse yet overlapping morphology,M. rothsteini(Forel) is representative of the greater challenge that exists inAustralianMonomoriumsystematics. Here we investigate species boundaries inM. rothsteiniusing a molecular phylogenetic framework to interpret the complex overlap of nine morphological characters (with 31 states) and examine biogeographic relationships among the lineages. Bayesian inference resolved 38 mtDNAlineages that were morphologically separable, at least from their sister lineage. Although the morphological characters were intermixed across the phylogeny, instances of inseparable morphology among sister clades was rare. Seventeen lineages exhibited complete morphological overlap with one or more other lineages and could not be separated byPrincipalComponentAnalysis based on 12 morphometric variables. Two‐thirds of all lineages occurred sympatrically with one or more both genetically and morphologically divergent lineages. The two nuclear markersEF1αF2andwinglesswere used to generate haplotype networks which were characterised by a star‐like pattern indicative of a rapid and recent radiation. Several haplotypes for both nuclear gene regions were shared among individuals occurring in separate mtDNAclades which we were also unable to distinguish morphologically or that were occurring in sympatry, indicating possible introgression in both the mtDNAand nuclear genomes. Clear biogeographic affinities among samples within a lineage were detected but there was no overall pattern in the biogeographic relationships among the lineages. We conclude thatM. rothsteiniis a large species complex that has undergone a complex evolutionary history following aridification of theAustralian continent, and discuss the implications of this conclusion for the systematics ofAustralianMonomoriummore generally.
The effect of C1‐inhibitor in a murine model of transfusion‐related acute lung injury
Authors: Müller M. C. A. , Stroo I. , Wouters D. , Zeerleder S. S. , Roelofs J. J. T. H. , Boon L. , Vroom M. B. , Juffermans N. P.
Published: December 2013

Background and objective

Transfusion‐related acute lung injury (TRALI) is the leading cause of transfusion‐related morbidity and mortality. Specific therapy is lacking. We assessed whether C1‐inhibitor attenuates lung injury in a ‘two‐hit’ TRALI model.

Methods

Mice were primed with lipopolysaccharide, subsequently TRALI was induced by MHC‐I antibodies. In the intervention group, C1‐inhibitor was infused concomitantly. Mice were supported with mechanical ventilation. After 2 h, mice were killed, lungs were removed and bronchoalveolar lavage fluid (BALF) was obtained.

Results

Injection of MHC‐I antibodies induced TRALI, illustrated by an increase in wet‐to‐dry ratio of the lungs, in BALF protein levels and in lung injury scores. TRALI was further characterized by complement activation, demonstrated by increased BALF levels of C3a and C5a. Administration of C1‐inhibitor resulted in increased pulmonary C1‐inhibitor levels with high activity. C1‐inhibitor reduced pulmonary levels of complement C3a associated with improved lung injury scores. However, levels of pro‐inflammatory mediators were unaffected.

Conclusion

In a murine model of TRALI, C1‐inhibitor attenuated pulmonary levels of C3a associated with improved lung injury scores, but with persistent high levels of inflammatory cytokines.