bluuebunny/crossref_metadata_2025 huggingface dataset:
p35 and Rac1 underlie the neuroprotection and cognitive improvement induced by CDK5 silencing
Authors: Posada‐Duque Rafael Andres , López‐Tobón Alejandro , Piedrahita Diego , González‐Billault Christian , Cardona‐Gomez Gloria Patricia
Published: April 2015

CDK5 plays an important role in neurotransmission and synaptic plasticity in the normal function of the adult brain, and dysregulation can lead to Tau hyperphosphorylation and cognitive impairment. In a previous study, we demonstrated that RNAi knock down of CDK5 reduced the formation of neurofibrillary tangles (NFT) and prevented neuronal loss in triple transgenic Alzheimer's mice. Here, we report that CDK5 RNAi protected against glutamate‐mediated excitotoxicity using primary hippocampal neurons transduced with adeno‐associated virus 2.5 viral vector eGFP‐tagged scrambled or CDK5 shRNA‐miR during 12 days. Protection was dependent on a concomitant increase in p35 and was reversed using p35 RNAi, which affected the down‐stream Rho GTPase activity. Furthermore, p35 over‐expression and constitutively active Rac1 mimicked CDK5 silencing‐induced neuroprotection. In addition, 3xTg‐Alzheimer's disease mice (24 months old) were injected in the hippocampus with scrambled or CDK5 shRNA‐miR, and spatial learning and memory were performed 3 weeks post‐injection using ‘Morris’ water maze test. Our data showed that CDK5 knock down induced an increase in p35 protein levels and Rac activity in triple transgenic Alzheimer's mice, which correlated with the recovery of cognitive function; these findings confirm that increased p35 and active Rac are involved in neuroprotection. In summary, our data suggest that p35 acts as a mediator of Rho GTPase activity and contributes to the neuroprotection induced by CDK5 RNAi.

image

CDK5 plays an important role in neurotransmission in the normal function of the adult brain, and dysregulation can lead to Tau hyperphosphorylation and cognitive impairment. Our findings suggest that p35/Rac1 signaling is critical in the CDK5 shRNAmiR‐induced neuroprotection against glutamate neurotoxicity and is also correlated with the recovery of cognitive function in 3xTg‐AD mice. CDK5 shRNAmiR blocks calpain activation, and the cleavage of p35 to p25 produced by glutamate, which generates neuroprotection in a p35 up‐regulation dependent mode and its down‐stream control of Rho GTPases, such as Rac1 and RhoA.
Effects of maternal dietary EPA and DHA supplementation and breeder age on embryonic and post‐hatch performance of broiler offspring
Authors: Koppenol A. , Delezie E. , Wang Y. , Franssens L. , Willems E. , Ampe B. , Buyse J. , Everaert N.
Published: April 2015

Breeder age and nutrition are amongst the most important factors affecting progeny growth and development. The present experiment was carried out to evaluate the effects of n‐3 fatty acid (FA), with special emphasis on the ratio of eicosapentaenoic (EPA, 20:5 n‐3) and docosahexaenoic (DHA, 22:6 n‐3) acid, provided to the diet of ageing broiler breeder hens at different ratios, on the incubation parameters and the performance of the offspring. Four hundred and eighty Ross‐308 broiler breeder hens were fed one of four different diets (120/treatment), with an equal fat content. The control diet was a basal diet, rich in n‐6 FAs (CON). Blends of fish oil were used to enrich the three other diets in n‐3 FA and to obtain different EPA/DHA ratios of 1/1 (EPA=DHA), 1/2 (DHA) or 2/1 (EPA). Every 5 weeks, incubation parameters were assessed. Every 15 weeks, offspring was reared until slaughter age on a standard diet. Breeder age affected almost all incubation and post‐hatch parameters, whereas n‐3 FA treatment only lowered egg weight (p < 0.0001) and consequently hatched chick weight (p < 0.0001). Supplementation of EPA resulted in a higher proportional liver weight (p = 0.0219) at hatch, a lower body weight up to 28 days post‐hatch (p = 0.0418), a lower daily weight gain (p = 0.0498) and a higher feed conversion ratio (p = 0.0395) during the starter period (p = 0.0498), resulting in a higher overall offspring feed conversion ratio (p = 0.0317) compared to the control diet. DHA supplementation, on the other hand, resulted in a lower residual yolk weight (p = 0.0220) and a higher overall offspring mortality (p = 0.0125). In conclusion, supplementation of n‐3 FA could not counter the adverse effect of breeder flock age, but did not harm incubation or improve post‐hatch performance, either. EPA and DHA affected offspring development differently during early post‐hatch life.
Behind the diversity: Ontogenies of radiate, disciform, and discoid capitula of Chrysanthemum and its allies
Authors: Ren Jia‐Bin , Guo Yan‐Ping
Published: April 2015

Asteraceae are distinguishable by their head‐like inflorescence called a capitulum. There are a variety of capitulum types due to different combinations of different composing florets. Morphological evolution of capitula must involve homoplasy under selective constraints. Therefore, a comparative examination of ontogenies of divergent capitula among phylogenetically close taxa is important to investigate the genetic and molecular bases of this trait. In the subtribe Artemisiinae, tribe Anthemideae, three closely related genera have different flower heads: Chrysanthemum with the radiate, Ajania with the disciform, and Stilpnolepis with the discoid capitula. Through observations using scanning electron microscopy, we examined the morphogenetic processes of flower heads of representative species of these genera. The morphological differentiation of the discoid from the radiate/disciform capitula occurred early in the floral initiation stage. The development of the ray/marginal florets of the radiate/disciform capitula lagged behind the outermost disc florets throughout the phases of floral initiation and floral organogenesis. Primordia of their 1–2 dorsal corolla lobes and all stamens ceased to grow soon after initiation. In contrast, all florets on a discoid capitulum developed equally in rates and modes into bisexual actinomorphic flowers. Differentiation between the radiate and the disciform capitula occurred late during the floral maturation stage. Our data provided evidence to the interpretation that uniformly acropetal development only occurs in species with homogamous flower heads, and to the idea that the timing of ontogenetic divergence between taxa is positively correlated with the phylogenetic distance of the taxa.
An evaluation of a new tool to aid judgements of credibility in the medico‐legal setting
Authors: Akehurst Lucy , Easton Simon , Fuller Emily , Drane Grace , Kuzmin Katherine , Litchfield Sarah
Published: April 2015

Purpose

Clinical psychologists and other health professionals are often requested to act as expert witnesses in Court. They are required to assess, and report upon, the reliability of the accounts of physical and psychological symptoms made by their clients. This study investigated the effectiveness of a checklist drawing upon relevant literature on lying and malingering to aid the detection of exaggeration of physical symptoms.

Method

Sixty‐four participants were cast as interviewers and assigned to either a ‘checklist’ or ‘no checklist’ condition. Another 64 volunteers were assigned to either a ‘truth teller’ or ‘malingerer’ role and, after undergoing a cold pressor procedure, were interviewed about their experience. The interviewers with a checklist drawn from the literature were asked to rate the presence of 28 checklist items on 5‐point Likert scales and to indicate whether or not they believed their interviewee was truthful or exaggerating his or her symptoms. The interviewers without the checklist were asked to simply indicate whether their interviewee was truthful or exaggerating.

Results

Evaluators who were not given the checklist did not classify their interviewees at a level significantly better than chance. Those using the checklist achieved an overall hit rate of 70%. Signal detection analysis supported the finding that those with the checklist showed greater discriminability. Nine checklist items significantly discriminated between truth tellers and malingerers. Furthermore, the total checklist score was significantly higher for exaggerated accounts than for truthful accounts.

Conclusions

Results suggest that a checklist based on the literatures into lying and malingering warrants further investigation. Such a tool would be useful as an aid for expert witnesses called to provide informed opinion on the likelihood that a claimant is exaggerating, malingering or otherwise misrepresenting difficulties.
3D superimposition and understanding temporomandibular joint arthritis
Authors: Cevidanes L. H. S. , Gomes L. R. , Jung B. T. , Gomes M. R. , Ruellas A. C. O. , Goncalves J. R. , Schilling J. , Styner M. , Nguyen T. , Kapila S. , Paniagua B.
Published: April 2015

Structured Abstract

Objectives

To investigate the 3D morphological variations in 169 temporomandibular ioint (TMJ) condyles, using novel imaging statistical modeling approaches.

Setting and sample population

The Department of Orthodontics and Pediatric Dentistry at the University of Michigan. Cone beam CT scans were acquired from 69 subjects with long‐term TMJ osteoarthritis (OA, mean age 39.1 ± 15.7 years), 15 subjects at initial consult diagnosis of OA (mean age 44.9 ±14.8 years), and seven healthy controls (mean age 43 ± 12.4 years).

Materials and methods

3D surface models of the condyles were constructed, and homologous correspondent points on each model were established. The statistical framework included Direction–Projection–Permutation (DiProPerm) for testing statistical significance of the differences between healthy controls and the OA groups determined by clinical and radiographic diagnoses.

Results

Condylar morphology in OA and healthy subjects varied widely with categorization from mild to severe bone degeneration or overgrowth. DiProPerm statistics supported a significant difference between the healthy control group and the initial diagnosis of OA group (t = 6.6, empirical p‐value = 0.006) and between healthy and long‐term diagnosis of OA group (t = 7.2, empirical p‐value = 0). Compared with healthy controls, the average condyle in OA subjects was significantly smaller in all dimensions, except its anterior surface, even in subjects with initial diagnosis of OA.

Conclusion

This new statistical modeling of condylar morphology allows the development of more targeted classifications of this condition than previously possible.
Genetics of the dentofacial variation in human malocclusion
Authors: Moreno Uribe L. M. , Miller S. F.
Published: April 2015

Malocclusions affect individuals worldwide, resulting in compromised function and esthetics. Understanding the etiological factors contributing to the variation in dentofacial morphology associated with malocclusions is the key to develop novel treatment approaches. Advances in dentofacial phenotyping, which is the comprehensive characterization of hard and soft tissue variation in the craniofacial complex, together with the acquisition of large‐scale genomic data have started to unravel genetic mechanisms underlying facial variation. Knowledge on the genetics of human malocclusion is limited even though results attained thus far are encouraging, with promising opportunities for future research. This review summarizes the most common dentofacial variations associated with malocclusions and reviews the current knowledge of the roles of genes in the development of malocclusions. Lastly, this review will describe ways to advance malocclusion research, following examples from the expanding fields of phenomics and genomic medicine, which aim to better patient outcomes.
Phenotype–genotype correlations of facial width and height proportions in patients with Class II malocclusion
Authors: Moreno Uribe L. M. , Ray A. , Blanchette D. R. , Dawson D. V. , Southard T. E.
Published: April 2015

Structured Abstract

Objectives

To characterize soft‐tissue facial height and width variation in Class II malocclusion and test for correlations with genes HMGA2, AJUBA, and ADK.

Setting and sample population

Nine facial proportions were estimated from 2D frontal repose photographs of 330 Caucasian adults with Class II malocclusion.

Material and methods

After adjustments for age and gender, the facial proportions were submitted to a principal component analyses (PCA). The most meaningful phenotypic variations were correlated with SNPs rs7924176 (ADK), rs17101923 (HMGA2), and rs997154 (AJUBA) genotyped in 106 individuals.

Results

Principal component analyses resulted in four principal components (PCs), which explained 75% of total variation. PC1 captured variation in the intercanthus distance and explained 28% of total variation. PC2 explained 21% of the variations in facial taper and facial index. PC3 explained 14% and reflected variations in the vertical dimension of the lower face. PC4 explained 12% and captured variations in distance between the eyes, width of the commissures, and the length of the superior aspect of the lower face height corresponding to the vertical dimension of the philtrum of the upper lip. A suggestive association (p < 0.05) was observed between PC4 and rs997154 corroborating the role of AJUBA in variation of facial dimensions.

Conclusion

2D frontal photographs can be used to derive quantitative measures of soft‐tissue phenotypes that are of clinical relevance. The methods described are suitable for discovery and replication of associations between genotypes and malocclusion phenotypes.
A pilot study of ambulatory masticatory muscle activities in temporomandibular joint disorders diagnostic groups
Authors: Iwasaki L. R. , Gonzalez Y. M. , Liu H. , Marx D. B. , Gallo L. M. , Nickel J. C.
Published: April 2015

Structured Abstract

Objective

To determine differences in masticatory muscle usage between temporomandibular joint disorders diagnostic groups.

Setting and sample population

Seventy‐one informed and consented subjects (27 men; 44 women) participated at the University at Buffalo.

Material and methods

Research diagnostic criteria and imaging data were used to categorize subjects according to the presence/absence (+/−) of TMJ disc placement (DD) and chronic pain (P) (+DD+P, n = 18; +DD−P, n = 14; −DD−P, n = 39). Electromyographic (EMG)/bite‐force calibrations determined subject‐specific masseter and temporalis muscle activities per 20 N bite‐force (T20N, μV). Over 3 days and nights, subjects collected EMG recordings. Duty factors (DFs, % of recording time) were determined based on threshold intervals (5–9, 10–24, 25–49, 50–79, ≥80% T20N). anova and Tukey–Kramer post hoc tests identified 1) diagnostic group differences in T20N and 2) the effects of diagnostic group, gender, time and interval on muscle DFs.

Results

Mean (± SE) temporalis T20N in +DD+P subjects was significantly higher (71.4 ± 8.8 μV) than masseter T20N in these subjects (19.6 ± 8.8 μV; p = 0.001) and in −DD−P subjects (25.3 ± 6.0 μV, p = 0.0007). Masseter DFs at 5–9% T20N were significantly higher in +DD−P women (3.48%) than +DD−P men (0.85%) and women and men in both other diagnostic groups (all p < 0.03), and in +DD+P women (2.00%) compared to −DD−P men (0.83%; p = 0.029). Night‐time DFs at 5–9% T20N in +DD−P women (1.97%) were significantly higher than in −DD−P men (0.47%) and women (0.24%; all p < 0.01).

Conclusions

Between‐group differences were found in masticatory muscle activities in both laboratory and natural environmental settings.
Prehospital Pain and Analgesic Therapy in Elderly Patients with Hip Fractures
Authors: Oberkircher Ludwig , Schubert Natalie , Eschbach Daphne‐Asimenia , Bliemel Christopher , Krueger Antonio , Ruchholtz Steffen , Buecking Benjamin
Published: April 2015

Introduction

As a part of aging, hip fractures are becoming more common. The connection between increased pain and a poor outcome has previously been shown. Therefore, even in prehospital situations, analgesic therapy appears to be reasonable. We established a prospective study with 153 patients to evaluate the patients' pain levels during the prehospital phase of treatment and prehospital analgesic therapy.

Methods

We performed a prospective study on 153 patients the age of 60 years or older in a University hospital setting between 2010 and 2011 who suffered hip fracture. Analgesics given and the type of medical staff that was involved were documented. Pain was measured using the NRS upon initial contact of the medical staff and upon admission to our emergency department.

Results

Initial pain level evaluated by EMS (emergency medical service) was 6.8 (SD = 2.7). Twenty‐two percent of the patients reported an NRS of 10 as the highest value following their injury. Forty‐three of 153 patients (28%) received analgesics. The mean initial pain score for those 43 patients who did receive pain medication was 7.0 (SD = 2.6). However, this score dropped to a mean of 2.8 (SD = 1.4) upon hospital arrival (P < 0.001). The patients who did not receive pain medication had an initial pain score of 4.5 (SD = 1.9). Upon admission to the hospital, this score decreased to a mean of 4.0 (SD = 1.7, P = 0.092).

Conclusion

Only a minority of patients with hip fractures received prehospital analgesia. The administration of prehospital analgesia was associated with significant pain relief.
Laryngoplasty in Standing Horses
Authors: Rossignol Fabrice , Vitte Amélie , Boening Josef , Maher Michael , Lechartier Antoine , Brandenberger Olivier , Martin‐Flores Manuel , Lang Hayley , Walker Wade , Ducharme Norm G.
Published: April 2015

Objective

To describe the clinical experience with standing laryngoplasty in a series of horses mostly nonracing.

Study Design

Case series.

Animals

Seventy‐one client‐owned horses.

Methods

Medical records (April 2008–February 2014) of horses treated by standing laryngoplasty for abnormal respiratory noise and or poor performance were reviewed. Horses were included if they had a diagnosis of idiopathic right or left recurrent laryngeal neuropathy confirmed by videoendoscopy. All horses underwent a unilateral laryngoplasty with a unilateral or bilateral ventriculectomy or ventriculocordectomy. Follow‐up endoscopy was performed in all horses within 24 hours postoperative, in 24 horses at 2‐weeks, and in 65 horses at 6 weeks. Late follow‐up was obtained from the trainer, owner, or referring veterinarian by telephone.

Results

Laryngoplasty was performed under endoscopic guidance with the horses sedated, and the surgical site was desensitized with local anesthetic solution. Laryngoplasty was completed in all horses and was well tolerated. No hyperabduction was observed. Two horses developed incisional swelling that resolved with drainage only. Late follow‐up reported satisfactory improvement in respiration in all but 3 horses.

Conclusions

Laryngoplasty performed with the horse standing avoids risks associated with general anesthesia and recovery and yields comparable results in nonracing horses, to laryngoplasty performed with the horse anesthetized. This technique reduces cost and allows accurate intraoperative adjustment of the degree of arytenoid abduction.
Robust optimization methods for cardiac sparing in tangential breast IMRT
Authors: Mahmoudzadeh Houra , Lee Jenny , Chan Timothy C. Y. , Purdie Thomas G.
Published: April 2015

Purpose:

In left‐sided tangential breast intensity modulated radiation therapy (IMRT), the heart may enter the radiation field and receive excessive radiation while the patient is breathing. The patient's breathing pattern is often irregular and unpredictable. We verify the clinical applicability of a heart‐sparing robust optimization approach for breast IMRT. We compare robust optimized plans with clinical plans at free‐breathing and clinical plans at deep inspiration breath‐hold (DIBH) using active breathing control (ABC).

Methods:

Eight patients were included in the study with each patient simulated using 4D‐CT. The 4D‐CT image acquisition generated ten breathing phase datasets. An average scan was constructed using all the phase datasets. Two of the eight patients were also imaged at breath‐hold using ABC. The 4D‐CT datasets were used to calculate the accumulated dose for robust optimized and clinical plans based on deformable registration. We generated a set of simulated breathing probability mass functions, which represent the fraction of time patients spend in different breathing phases. The robust optimization method was applied to each patient using a set of dose‐influence matrices extracted from the 4D‐CT data and a model of the breathing motion uncertainty. The goal of the optimization models was to minimize the dose to the heart while ensuring dose constraints on the target were achieved under breathing motion uncertainty.

Results:

Robust optimized plans were improved or equivalent to the clinical plans in terms of heart sparing for all patients studied. The robust method reduced the accumulated heart dose (D10cc) by up to 801 cGy compared to the clinical method while also improving the coverage of the accumulated whole breast target volume. On average, the robust method reduced the heart dose (D10cc) by 364 cGy and improved the optBreast dose (D99%) by 477 cGy. In addition, the robust method had smaller deviations from the planned dose to the accumulated dose. The deviation of the accumulated dose from the planned dose for the optBreast (D99%) was 12 cGy for robust versus 445 cGy for clinical. The deviation for the heart (D10cc) was 41 cGy for robust and 320 cGy for clinical.

Conclusions:

The robust optimization approach can reduce heart dose compared to the clinical method at free‐breathing and can potentially reduce the need for breath‐hold techniques.
Reducing the effects of acoustic heterogeneity with an iterative reconstruction method from experimental data in microwave induced thermoacoustic tomography
Authors: Wang Jinguo , Zhao Zhiqin , Song Jian , Chen Guoping , Nie Zaiping , Liu Qing‐Huo
Published: April 2015

Purpose:

An iterative reconstruction method has been previously reported by the authors of this paper. However, the iterative reconstruction method was demonstrated by solely using the numerical simulations. It is essential to apply the iterative reconstruction method to practice conditions. The objective of this work is to validate the capability of the iterative reconstruction method for reducing the effects of acoustic heterogeneity with the experimental data in microwave induced thermoacoustic tomography.

Methods:

Most existing reconstruction methods need to combine the ultrasonic measurement technology to quantitatively measure the velocity distribution of heterogeneity, which increases the system complexity. Different to existing reconstruction methods, the iterative reconstruction method combines time reversal mirror technique, fast marching method, and simultaneous algebraic reconstruction technique to iteratively estimate the velocity distribution of heterogeneous tissue by solely using the measured data. Then, the estimated velocity distribution is used subsequently to reconstruct the highly accurate image of microwave absorption distribution. Experiments that a target placed in an acoustic heterogeneous environment are performed to validate the iterative reconstruction method.

Results:

By using the estimated velocity distribution, the target in an acoustic heterogeneous environment can be reconstructed with better shape and higher image contrast than targets that are reconstructed with a homogeneous velocity distribution.

Conclusions:

The distortions caused by the acoustic heterogeneity can be efficiently corrected by utilizing the velocity distribution estimated by the iterative reconstruction method. The advantage of the iterative reconstruction method over the existing correction methods is that it is successful in improving the quality of the image of microwave absorption distribution without increasing the system complexity.
Genetic Diversity of the
fliC
Genes Encoding the Flagellar Antigen H19 of Escherichia coli and Application to the Specific Identification of Enterohemorrhagic E. coli O121:H19
Authors: Beutin Lothar , Delannoy Sabine , Fach Patrick
Published: April 2015

Enterohemorrhagic
Escherichia coli
(EHEC) O121:H19 belong to a specific clonal type distinct from other classical EHEC and major enteropathogenic
E. coli
groups and is regarded as one of the major EHEC serogroups involved in severe infections in humans. Sequencing of the
fliC
genes associated with the flagellar antigen H19 (
fliC
H19
) revealed the genetic diversity of the
fliC
H19
gene sequences in
E. coli
. A cluster analysis of 12
fliC
H19
sequences, 4 from O121 and 8 from non-O121
E. coli
strains, revealed five different genotypes. All O121:H19 strains fell into one cluster, whereas a second cluster was formed by five non-O121:H19 strains. Cluster 1 and cluster 2 strains differ by 27 single nucleotide exchanges in their
fliC
H19
genes (98.5% homology). Based on allele discrimination of the
fliC
H19
genes, a real-time PCR test was designed for specific identification of EHEC O121:H19. The O121
fliC
H19
PCR tested negative in 73
E. coli
H19 strains that belonged to serogroups other than O121, including 28 different O groups, O-nontypeable H19, and O-rough:H19 strains. The O121
fliC
H19
PCR reacted with all 16 tested O121:H19 strains and 1 O-rough:H19 strain which was positive for the O121
wzx
gene. A cross-reaction was observed only with
E. coli
H32 strains which share sequence similarities in the target region of the O121
fliC
H19
PCR. The combined use of O-antigen genotyping (O121
wzx
) and the detection of O121
fliC
H19
allele type contributes to improving the identification and molecular serotyping of EHEC O121:H19 motile and nonmotile strains and variants of these strains lacking
stx
genes.
Steroid Hydroxylation by Basidiomycete Peroxygenases: a Combined Experimental and Computational Study
Authors: Babot Esteban D. , del Río José C. , Cañellas Marina , Sancho Ferran , Lucas Fátima , Guallar Víctor , Kalum Lisbeth , Lund Henrik , Gröbe Glenn , Scheibner Katrin , Ullrich René , Hofrichter Martin , Martínez Angel T. , Gutiérrez Ana
Published: April 2015

The goal of this study is the selective oxyfunctionalization of steroids under mild and environmentally friendly conditions using fungal enzymes. With this purpose, peroxygenases from three basidiomycete species were tested for the hydroxylation of a variety of steroidal compounds, using H
2
O
2
as the only cosubstrate. Two of them are wild-type enzymes from
Agrocybe aegerita
and
Marasmius rotula
, and the third one is a recombinant enzyme from
Coprinopsis cinerea
. The enzymatic reactions on free and esterified sterols, steroid hydrocarbons, and ketones were monitored by gas chromatography, and the products were identified by mass spectrometry. Hydroxylation at the side chain over the steroidal rings was preferred, with the 25-hydroxyderivatives predominating. Interestingly, antiviral and other biological activities of 25-hydroxycholesterol have been reported recently (M. Blanc et al., Immunity 38:106–118, 2013,
http://dx.doi.org/10.1016/j.immuni.2012.11.004
). However, hydroxylation in the ring moiety and terminal hydroxylation at the side chain also was observed in some steroids, the former favored by the absence of oxygenated groups at C-3 and by the presence of conjugated double bonds in the rings. To understand the yield and selectivity differences between the different steroids, a computational study was performed using Protein Energy Landscape Exploration (PELE) software for dynamic ligand diffusion. These simulations showed that the active-site geometry and hydrophobicity favors the entrance of the steroid side chain, while the entrance of the ring is energetically penalized. Also, a direct correlation between the conversion rate and the side chain entrance ratio could be established that explains the various reaction yields observed.
Structure and Mechanism of Ferulic Acid Decarboxylase (FDC1) from Saccharomyces cerevisiae
Authors: Bhuiya Mohammad Wadud , Lee Soon Goo , Jez Joseph M. , Yu Oliver
Published: April 2015

The nonoxidative decarboxylation of aromatic acids occurs in a range of microbes and is of interest for bioprocessing and metabolic engineering. Although phenolic acid decarboxylases provide useful tools for bioindustrial applications, the molecular bases for how these enzymes function are only beginning to be examined. Here we present the 2.35-Å-resolution X-ray crystal structure of the ferulic acid decarboxylase (FDC1; UbiD) from
Saccharomyces cerevisiae
. FDC1 shares structural similarity with the UbiD family of enzymes that are involved in ubiquinone biosynthesis. The position of 4-vinylphenol, the product of
p
-coumaric acid decarboxylation, in the structure identifies a large hydrophobic cavity as the active site. Differences in the β2e-α5 loop of chains in the crystal structure suggest that the conformational flexibility of this loop allows access to the active site. The structure also implicates Glu285 as the general base in the nonoxidative decarboxylation reaction catalyzed by FDC1. Biochemical analysis showed a loss of enzymatic activity in the E285A mutant. Modeling of 3-methoxy-4-hydroxy-5-decaprenylbenzoate, a partial structure of the physiological UbiD substrate, in the binding site suggests that an ∼30-Å-long pocket adjacent to the catalytic site may accommodate the isoprenoid tail of the substrate needed for ubiquinone biosynthesis in yeast. The three-dimensional structure of yeast FDC1 provides a template for guiding protein engineering studies aimed at optimizing the efficiency of aromatic acid decarboxylation reactions in bioindustrial applications.
Developmental Growth Control Exerted via the Protein A Kinase Tpk2 in Ashbya gossypii
Authors: Wasserstrom Lisa , Lengeler Klaus , Walther Andrea , Wendland Jürgen
Published: April 2015

Sporulation in
Ashbya gossypii
is induced by nutrient-limited conditions and leads to the formation of haploid spores. Using RNA-seq, we have determined a gene set induced upon sporulation, which bears considerable overlap with that of
Saccharomyces cerevisiae
but also contains
A. gossypii
-specific genes. Addition of cyclic AMP (cAMP) to nutrient-limited media blocks sporulation and represses the induction of sporulation specific genes. Deletion of the protein kinase A (PKA) catalytic subunits encoded by
TPK1
and
TPK2
showed reduced growth in
tpk1
but enhanced growth in the
tpk2
strain; however, both mutants sporulated well. Sporulation can be blocked by cAMP in
tpk1
but not in
tpk2
strains. Similarly,
TPK2
acts at a second developmental switch promoting the break in spore dormancy. In
S. cerevisiae
, PKA phosphorylates and inhibits Msn2/4. The transcript profiles of the
tpk1
and
msn2/4
mutants were very similar to that of the wild type under sporulation conditions. However, deletion of the single
A. gossypii
MSN2/4
homolog generated a specific sporulation defect. We identified a set of genes involved in spore wall assembly that was downregulated in the
msn2/4
mutant, particularly
DIT2
, suggesting that poor spore viability may be due to lysis of spores. Our results reveal specific functional differences between the two catalytic PKA subunits in
A. gossypii
and identified Tpk2 as the key A kinase that transduces developmental decisions of growth. Our data also suggest that Msn2/4 is involved only at a late step of sporulation in
A. gossypii
and is not a major regulator of
IME1
.
Genetic variation at the human connexin 43 locus but not at the connexin 40 locus is associated with left bundle branch block
Authors: Ladenvall Per , Andersson Björn , Dellborg Mikael , Hansson Per-Olof , Eriksson Henry , Thelle Dag , Eriksson Peter
Published: April 2015

Background

Bundle branch block (BBB) has been regarded as a disease of the conduction system, but occurs in mice lacking connexin 40 (expressed in atria, proximal conduction system) or connexin 43 (expressed in Purkinje cells, cardiomyocytes).

Objective

The aim of this paper is to explore whether BBB is heritable, and whether polymorphisms at connexin 40 and connexin 43 loci are associated with BBB.

Methods

To assess BBB heritability, we screened descendants of men with BBB in the population cohort ‘The Study of Men Born 1913’. DNA samples from 80-year-old men with extreme QRS-duration phenotypes were used to search for polymorphisms at connexin 40 and 43 loci. Associations between identified polymorphisms and BBB were evaluated in an independent cohort (INTERGENE).

Results

Seventy-seven men from ‘The Study of Men Born 1913’ with BBB had 116 descendants. Among the 76 participating descendants, 2 sons (6.4%) had BBB at 54 years of age. At the same age, 0.9% of men born in 1913 had BBB. We identified 6 single nucleotide polymorphisms (SNPs) in connexin 40 and 1 polymorphism in connexin 43. In the INTERGENE cohort, the connexin 43 polymorphism was associated with left BBB (LBBB) (4 of 35 LBBB vs 16 of 232 without BBB, χ2=7.4, p=0.03), but not with right BBB (RBBB) or overall BBB. None of the connexin 40 SNPs or haplotypes were associated with LBBB or RBBB.

Conclusions

These findings indicate that conduction by connexin 43 within the ventricular muscle distal to the specialised conduction system may be important for LBBB development.
The Immunobiology of Interleukin-27
Authors: Yoshida Hiroki , Hunter Christopher A.
Published: April 2015

Interleukin-27 (IL-27) is a cytokine with strikingly diverse influences on the immune response. Although it was initially linked with the development of Th1 responses, it is now recognized as a potent antagonist of different classes of inflammation through its ability to directly modify CD4+and CD8+T cell effector functions, to induce IL-10, and to promote specialized T regulatory cell responses. Although this aspect of IL-27 biology has provided insights into how the immune system prevents hyperactivity in the setting of infectious and autoimmune inflammation, in vaccination and cancer models the stimulatory effects of IL-27 on CD8+T cell function appear prominent. Additionally, associations between IL-27 and antibody-mediated disease have led to an interest in defining the impact of IL-27 on innate immunity and humoral responses in different disease states. The maturation of this literature has been accompanied by attempts to translate these findings from experimental models into human diseases and by efforts to define where IL-27 might represent a viable therapeutic target.
Tetrandrine reverses human cardiac myofibroblast activation and myocardial fibrosis
Authors: Teng Guoqi , Svystonyuk Daniyil , Mewhort Holly E. M. , Turnbull Jeannine D. , Belke Darrell D. , Duff Henry J. , Fedak Paul W. M.
Published: April 2015

Tetrandrine (TTD) is a calcium channel blocker with documented antifibrotic actions. In this study, for the first time, we identified that TTD can directly prevent in vitro human cardiac myofibroblast activation and limit in vivo myocardial fibrosis. In vitro, cardiac myofibroblasts from human atrial biopsies ( N = 10) were seeded in three-dimensional collagen matrices. Cell-collagen constructs were exposed to transforming growth factor-β1(10 ng/ml), with or without TTD (1 and 5 μM) for 48 h. Collagen gel contraction, myofibroblast activation (α-smooth muscle actin expression), expression of profibrotic mRNAs, and rate of collagen protein synthesis were compared. TTD decreased collagen gel contraction (79.7 ± 1.3 vs 60.1 ± 8.9%, P < 0.01), α-smooth muscle actin expression (flow cytometry), collagen synthesis ([3H]proline incorporation), and collagen mRNA expression. Cell viability was similar between groups (annexin positive cells: 1.7 vs. 1.4%). TTD inhibited collagen gel contraction in the presence of T-type and L-type calcium channel blockers, and the intracellular calcium chelator BAPTA-AM (15 μM), suggesting that the observed effects are not mediated by calcium homeostasis. In vivo, Dahl salt-sensitive hypertensive rats were treated with variable doses of TTD (by intraperitoneal injection over 4 wk) and compared with untreated controls ( N = 12). Systemic blood pressure was monitored by tail cuff. Myocardial fibrosis and left ventricular compliance were assessed by histology and passive pressure-volume analysis. Myocardial fibrosis was attenuated compared with untreated controls (%collagen area: 9.4 ± 7.3 vs 2.1 ± 1.0%, P < 0.01). Left ventricular compliance was preserved. In conclusion, TTD reverses human cardiac myofibroblast activation and myocardial fibrosis, independent of calcium channel blockade.
Right ventricular cyclic nucleotide signaling is decreased in hyperoxia-induced pulmonary hypertension in neonatal mice
Authors: Heilman Rachel P. , Lagoski Megan B. , Lee Keng Jin , Taylor Joann M. , Kim Gina A. , Berkelhamer Sara K. , Steinhorn Robin H. , Farrow Kathryn N.
Published: April 2015

Pulmonary hypertension (PH) and right ventricular hypertrophy (RVH) affect 25–35% of premature infants with significant bronchopulmonary dysplasia (BPD), increasing morbidity and mortality. We sought to determine the role of phosphodiesterase 5 (PDE5) in the right ventricle (RV) and left ventricle (LV) in a hyperoxia-induced neonatal mouse model of PH and RVH. After birth, C57BL/6 mice were placed in room air (RA) or 75% O2(CH) for 14 days to induce PH and RVH. Mice were euthanized at 14 days or recovered in RA for 14 days or 42 days prior to euthanasia at 28 or 56 days of age. Some pups received sildenafil or vehicle (3 mg·kg−1·dose−1sc) every other day from P0. RVH was assessed by Fulton's index [RV wt/(LV + septum) wt]. PDE5 protein expression was analyzed via Western blot, PDE5 activity was measured by commercially available assay, and cGMP was measured by enzyme-linked immunoassay. Hyperoxia induced RVH in mice after 14 days, and RVH did not resolve until 56 days of age. Hyperoxia increased PDE5 expression and activity in RV, but not LV + S, after 14 days. PDE5 expression normalized by 28 days of age, but PDE5 activity did not normalize until 56 days of age. Sildenafil given during hyperoxia prevented RVH, decreased RV PDE5 activity, and increased RV cGMP levels. Mice with cardiac-specific overexpression of PDE5 had increased RVH in RA. These findings suggest normal RV PDE5 function is disrupted by hyperoxia, and elevated PDE5 contributes to RVH and remodeling. Therefore, in addition to impacting the pulmonary vasculature, sildenafil also targets PDE5 in the neonatal mouse RV and decreases RVH.
An Antibody–Drug Conjugate Directed against Lymphocyte Antigen 6 Complex, Locus E (LY6E) Provides Robust Tumor Killing in a Wide Range of Solid Tumor Malignancies
Authors: Asundi Jyoti , Crocker Lisa , Tremayne Jarrod , Chang Peter , Sakanaka Chie , Tanguay Josh , Spencer Susan , Chalasani Sreedevi , Luis Elizabeth , Gascoigne Karen , Desai Rupal , Raja Rajiv , Friedman Brad A. , Haverty Peter M. , Polakis Paul , Firestein Ron
Published: April 2015

Purpose: Chemotherapies are limited by a narrow therapeutic index resulting in suboptimal exposure of the tumor to the drug and acquired tumor resistance. One approach to overcome this is through antibody–drug conjugates (ADC) that facilitate greater potency via target-specific delivery of highly potent cytotoxic agents.

Experimental Design: In this study, we used a bioinformatics approach to identify the lymphocyte antigen 6 complex locus E (LY6E), an IFN-inducible glycosylphosphatidylinositol (GPI)-linked cell membrane protein as a promising ADC target. We developed a monoclonal anti-LY6E antibody and characterized in situ LY6E expression in over 750 cancer specimens and normal tissues. Target-dependent anti-LY6E ADC killing was investigated both in vitro and in vivo using patient-derived xenograft models.

Results: Using in silico approaches, we found that LY6E was significantly overexpressed and amplified in a wide array of different human solid tumors. IHC analysis revealed high LY6E protein expression in a number of tumor types, such as breast, lung, gastric, ovarian, pancreatic, kidney and head/neck carcinomas. Characterization of the endocytic pathways for LY6E revealed that the LY6E-specific antibody is internalized into cells leading to lysosomal accumulation. Consistent with this, a LY6E-specific ADC inhibited in vitro cell proliferation and produced durable tumor regression in vivo in clinically relevant LY6E-expressing xenograft models.

Conclusions: Our results identify LY6E as a highly promising molecular ADC target for a variety of solid tumor types with current unmet medical need. Clin Cancer Res; 21(14); 3252–62. ©2015 AACR.
The Relationship between Age-Related Macular Degeneration and Olfactory Function
Authors: Kar Taner , Yildirim Yildiray , Altundağ Aytuğ , Sonmez Murat , Kaya Abdullah , Colakoglu Kadir , Tekeli Hakan , Cayonu Melih , Hummel Thomas
Published: April 2015

Background: Olfactory dysfunction is a common symptom of many neurodegenerative diseases, and age-related macular degeneration (AMD) is a late-onset neurodegenerative disease. Objective: Thus, the aim of this study was to investigate olfactory functions in patients with AMD. Methods: A total of 69 subjects with AMD and 69 age- and sex-matched healthy controls were enrolled. After a complete ophthalmic evaluation, the AMD patients were subclassified as early- and late-stage AMD. Psychophysical testing of olfactory function was performed using the validated Sniffin' Sticks test. Results: This study was carried out in 138 subjects, with a mean age of 74.3 ± 8.9 years (range 51-89). The current investigation showed the following two major findings: (1) patients with AMD had decreased olfactory abilities, especially in odor discrimination and odor identification, even at early stages compared to controls, whereas patients had decreased olfactory abilities in all subtasks of olfactory testings in advanced stages of AMD disease, and (2) as the visual acuity of AMD patients decreased, the olfactory abilities of these patients worsened. Conclusion: This study demonstrated that AMD had significant negative effects on all orthonasal olfactory tasks, particularly in advanced stages. Similar to other neurodegenerative diseases, odor discrimination and identification seemed to be more affected than odor detection threshold tasks.
Plasma rico em plaquetas autólogas na cicatrização do tendão do músculo gastrocnêmio de coelhos
Authors: Eurides Duvaldo , Guimarães Camila Pereira de Almeida , Beletti Marcelo Emílio , Mundim Antônio Vicente , Souza Luiz Augusto De , Gonçalves Gentil Ferreira , Eurides Gustavo Prudente
Published: April 2015

As lesões tendíneas podem envolver a secção parcial ou total do tendão calcâneo comum e ocasionar alterações posturais do membro locomotor. O presente trabalho avaliou o efeito da aplicação tópica de concentrado de plaquetas autólogas na reparação do tendão do musculo gastrocnêmio de coelhos, decorridos 45 e 90 dias de período pós-operatório (PO). Doze coelhos adultos foram separados em dois grupos (n = 6) e submetidos a punção cardíaca e colheita de 10 mL de sangue para obtenção do plasma rico em plaquetas (PRP). Nos animais de ambos os grupos, foi realizada a tenotomia transversal total no terço médio do ventre lateral do tendão do musculo gastrocnemio e a sua aproximação com a sutura de Kessler modificada e fio de nailon. Nos animais do grupo tratado, foi aplicado sobre a síntese dos tendões em media 490.644 mil plaquetas/μL do PRP e notou-se maior quantidade de fibras colágenas em relação ao controle, sendo que aos 90 dias de PO a intensidade de colágeno observado foi maior que nos animais com 45 dias, com maior quantidade de fibroblastos nos controles em relação aos tratados. A administração de concentrado de plasma de plaquetas autógenas no reparo do tendão gastrocnemio de coelhos estimula e organiza o processo de reparação e ocasiona produção precoce de fibras colágenas.
Editorial
Authors: Vasconcellos Silvio Arruda , Gennari Solange Maria
Published: April 2015

Apresentamos o primeiro número do Brazilian Journal of Veterinary Research and Animal Science - BJVRAS do ano de 2015. Neste ano importantes decisões foram tomadas: 1) visando propiciar a internacionalização da revista, a partir de março de 2015 só serão aceitos trabalhos apresentados na língua inglesa; 2) a composição dos membros externos à Universidade de São Paulo, que ocupam a posição de Editores Associados da revista foi alterada, os novos participantes, Prof. Dr. Júlio Lopes Sequeira da Universidade Estadual Paulista, campus de Botucatu, Prof. Dr. Sidnei Miyoshi Sakamoto da Universidade Federal do Semi-Árido, campus de Mossoró, Prof. Dr. Milton Ricardo Azedo da Universidade Metropolitana de Santos e Prof. Dr. José Luiz Laus da Universidade Estadual Paulista, campus de Jaboticabal passaram a fazer parte, respectivamente, das áreas de patologia, medicina veterinária preventiva, clínica veterinária e cirurgia veterinária. Agradecendo a colaboração dos Editores Associados anteriores que muito contribuíram para o fortalecimento do BJVRAS, estamos seguros que os ingressantes irão adotar a mesma dedicação e esforço necessários à manutenção da qualidade científica da revista; 3) as normas da revista foram revisadas e foi introduzida a possibilidade da publicação de relatos de casos. No presente número são publicados dois trabalhos da área de ciências básicas: um tratando da vascularização renal de equinos Puro Sangue Inglês e outro analisando aspectos da relação entre pontos anatômicos do sistema nervoso central e os sintomas mentais considerados na homeopatia veterinária. Na área de cirurgia são apresentados quatro trabalhos, três experimentais investigando possibilidades da evolução pós-cirúrgica com o emprego de pericárdio homólogo, de plaquetas ou do extrato de frutos da planta Jucá (Lidibia ferrea) e um de casuísmo compilando os atendimentos efetuados pelo setor de cirurgia do Hospital da Faculdade de Medicina Veterinária e Zootecnia da USP no período de 1988 a 2007. Na área de patologia, constam dois trabalhos relacionados à aquicultura, um experimental avaliando o efeito da temperatura na infecção de tilápias do Nilo pelo Streptococcus agalactiae e outro relatando um surto de mortalidade por Edwardsiella tarda em tambacús. A área de reprodução conta com dois trabalhos: um que analisa o efeito do sombreamento na eficiência reprodutora de touros Brangus e um que compara os métodos empregados para a incorporação de DNA exógeno pelos espermatozoides de bovinos. Desejamos a todos uma boa leitura.