bluuebunny/crossref_metadata_2025 huggingface dataset:
Forward chemical genetic screens in Arabidopsis identify genes that influence sensitivity to the phytotoxic compound sulfamethoxazole
Authors: Schreiber Karl J , Austin Ryan S , Gong Yunchen , Zhang Jianfeng , Fung Pauline , Wang Pauline W , Guttman David S , Desveaux Darrell
Published: November 2012

Background

The sulfanilamide family comprises a clinically important group of antimicrobial compounds which also display bioactivity in plants. While there is evidence that sulfanilamides inhibit folate biosynthesis in both bacteria and plants, the complete network of plant responses to these compounds remains to be characterized. As such, we initiated two forward genetic screens in Arabidopsis in order to identify mutants that exhibit altered sensitivity to sulfanilamide compounds. These screens were based on the growth phenotype of seedlings germinated in the presence of the compound sulfamethoxazole (Smex).

Results

We identified a mutant with reduced sensitivity to Smex, and subsequent mapping indicated that a gene encoding 5-oxoprolinase was responsible for this phenotype. A mutation causing enhanced sensitivity to Smex was mapped to a gene lacking any functional annotation.

Conclusions

The genes identified through our forward genetic screens represent novel mediators of Arabidopsis responses to sulfanilamides and suggest that these responses extend beyond the perturbation of folate biosynthesis.
Spreading brain lesions in a familial Creutzfeldt-Jakob disease with V180I mutation over 4 years
Authors: Deguchi Kentaro , Takamiya Motonori , Deguchi Shoko , Morimoto Nobutoshi , Kurata Tomoko , Ikeda Yoshio , Abe Koji
Published: November 2012

Background

We report a female patient with familial Creutzfeldt-Jakob disease with V180I mutation (fCJD with V180I), who was serially followed up with magnetic resonance imaging (MRI) and electroencephalogram (EEG) for up to four years.

Case presentation

At 6 months after the onset, diffusion-weighted images (DWI) and fluid-attenuated inversion recovery (FLAIR) of brain MRI revealed an increased signal intensity in the bilateral frontal, temporal, and parietal cerebral cortex with left dominancy except for the occipital lobe. However, her follow-up MRI at four years showed the high-signal regions spreading to the occipital cerebral cortex in DWI and FLAIR images, and bilateral frontal cerebral white matter in FLAIR images. EEG showed a progressive and general slow high-voltage rhythm from 7–8 to 3–5 c/s over four years, without evidence of periodic synchronous discharge. These findings correspond to the symptom progression even after akinetic mutism at 18 months.

Conclusion

We suggest that serial MRI and EEG examinations are useful for early diagnosis of fCJD with V180I and for monitoring disease progression.
Over prescription of antibiotics for adult pharyngitis is prevalent in developing countries but can be reduced using McIsaac modification of Centor scores: a cross-sectional study
Authors: Palla Amber Hanif , Khan Rafeeq Alam , Gilani Anwar H , Marra Fawziah
Published: November 2012

Background

AlthoughGroup A beta hemolytic streptococcus (GABHS)can cause bacterial pharyngitis, the most common etiology is viral; despite this viral etiology, antibiotics are commonly prescribed for this infection in industrialized countries. We investigated the prevalence ofGABHSin adult pharyngitis patients from lower socioeconomic settings in Karachi, Pakistan, how often antibiotics are prescribed for pharyngitis and if appropriate agents were used in a developing world setting. Finally, we wanted to see the usefulness of modified McIsaac scores in predicting positive cultures.

Methods

Adult patients were recruited from three local hospital outpatient dispensaries (OPDs). All patients aged 14–65 years who were suspected of having bacterial pharyngitis had throat swabs taken. Laboratory results forGABHSpharyngitis were then compared with their prescriptions. Appropriateness (using the World Health Organization’s definition) and type of antibiotic prescribed were assessed.

Results

Of 137 patients,30 patients each were studied for scores of 0, 1, 2 and 3; 17 patients were studied for score 4. Although 6 (4.4%) patients wereGABHS+, for a prevalence of 43.8 per 1000 population, antibiotics were prescribed to 135 patients (98.5%). Of these, only 11.1% received appropriate antibiotics while 88.9%received inappropriate antibiotics.Penicillins were prescribed most (34.1%), especially amoxicillin/clavulanate; followed by macrolides (31.1%), especially the second-generation agents, and fluoroquinolones (14.8%). McIsaac scores were found to be 100% sensitive and 68.7% specific, giving a positive predictive value (PPV) of 12.7% and a negative predictive value (NPV) of 100%.

Conclusions

Antibiotics were prescribed irrationally to adult pharyngitis patients, as most cultures were negative for bacterial infection. McIsaac modification of Centor scores related directly to culture results. We would therefore highly recommend its use to help family physicians make treatment decisions for adult pharyngitis patients.
Strengthening and stretching for rheumatoid arthritis of the hand (SARAH): design of a randomised controlled trial of a hand and upper limb exercise intervention - ISRCTN89936343
Authors:   Sarah Trial Team, Adams Jo , Bridle Chris , Dosanjh Sukhdeep , Heine Peter , Lamb Sarah E , Lord Joanne , McConkey Christopher , Nichols Vivien , Toye Francine , Underwood Martin R , Williams Mark A , Williamson Esther M
Published: November 2012

Background

Rheumatoid Arthritis (RA) commonly affects the hands and wrists with inflammation, deformity, pain, weakness and restricted mobility leading to reduced function. The effectiveness of exercise for RA hands is uncertain, although evidence from small scale studies is promising. The Strengthening And Stretching for Rheumatoid Arthritis of the Hand (SARAH) trial is a pragmatic, multi-centre randomised controlled trial evaluating the clinical and cost effectiveness of adding an optimised exercise programme for hands and upper limbs to best practice usual care for patients with RA.

Methods/design

480 participants with problematic RA hands will be recruited through 17 NHS trusts. Treatments will be provided by physiotherapists and occupational therapists. Participants will be individually randomised to receive either best practice usual care (joint protection advice, general exercise advice, functional splinting and assistive devices) or best practice usual care supplemented with an individualised exercise programme of strengthening and stretching exercises. The study assessors will be blinded to treatment allocation and will follow participants up at four and 12 months. The primary outcome measure is the Hand function subscale of the Michigan Hand Outcome Questionnaire, and secondary outcomes include hand and wrist impairment measures, quality of life, and resource use. Economic and qualitative studies will also be carried out in parallel.

Discussion

This paper describes the design and development of a trial protocol of a complex intervention study based in therapy out-patient departments. The findings will provide evidence to support or refute the use of an optimised exercise programme for RA of the hand in addition to best practice usual care.

Trial registration

Current Controlled Trials ISRCTN89936343
FSH prevents depletion of the resting follicle pool by promoting follicular number and morphology in fresh and cryopreserved primate ovarian tissues following xenografting
Authors: von Schönfeldt Viktoria , Chandolia Ramesh , Ochsenkühn Robert , Nieschlag Eberhard , Kiesel Ludwig , Sonntag Barbara
Published: November 2012

Background

Cryopreservation and transplantation of ovarian tissue is one option for re-establishing ovarian function, but optimal conditions for graft sustainment and follicular survival are still considered experimental. The present study aims to analyze the effect of FSH treatment on the resting follicle pool in fresh and cryopreserved primate ovarian tissues following xenografting.

Methods

Ovarian tissues from adult marmosets were grafted freshly or following cryopreservation to ovarectomized nude mice treated with FSH 25 IU twice daily post transplantation or left untreated as controls. Grafts were retrieved 2 or 4 weeks after transplantation to evaluate the number and morphological appearance of follicles.

Results

Early start of FSH treatment within 1 week following transplantation partly prevents primordial follicle loss in fresh and frozen-thawed tissues, whereas after a 3 weeks time interval this effect is present only in fresh tissues. A similar positive effect of early, but not later FSH treatment on primary follicles is seen in fresh tissues compared to only marginal effects in frozen-thawed tissues. The percentage of morphologically normal follicles is generally increased in FSH treated tissues, whereas the percentage of primary follicles over all primordial and primary follicles is increased by FSH only in freshly-grafted tissues.

Conclusions

FSH treatment alleviates depletion of the resting follicle pool and promotes normal follicular morphology both in freshly and frozen-thawed grafted tissues. In previously cryopreserved tissues, applying to most of the tissues intended for clinical use in fertility preservation attempts, its positive effect on primordial follicle numbers and potential graft sustainment is dependent on an early start of treatment within one week of transplantation.
Association of genome variations in the renin-angiotensin system with physical performance
Authors: Sgourou Argyro , Fotopoulos Vassilis , Kontos Vassilis , Patrinos George P , Papachatzopoulou Adamantia
Published: November 2012

Background

The aim of this study was to determine the genotype distribution and allelic frequencies of ACE (I/D), AGTR1 (A +1166 C), BDKRB2 (+9/−9) and LEP (G–2548A) genomic variations in 175 Greek athletes who excelled at a national and/or international level and 169 healthy Greek adults to identify whether some particular combinations of these loci might serve as predictive markers for superior physical condition.

Results

The D/D genotype of the ACE gene (p = 0.034) combined with the simultaneous existence of BDKRB2 (+9/−9) (p = 0.001) or LEP (G/A) (p = 0.021) genotypes was the most prevalent among female athletes compared to female controls. A statistical trend was also observed in BDKRB2 (+9/−9) and LEP (G–2548A) heterozygous genotypes among male and female Greek athletes, and in ACE (I/D) only in male athletes. Finally, both male and female athletes showed the highest rates in the AGTR1 (A/A) genotype.

Conclusions

Our results suggest that the co-existence of ACE (D/D), BDKRB2 (+9/−9) or LEP (G/A) genotypes in female athletes might be correlated with a superior level of physical performance.
Multilayer porous silicon diffraction gratings operating in the infrared
Authors: Lai Meifang , Sridharan Gayathri M , Parish Giacinta , Bhattacharya Shanti , Keating Adrian
Published: November 2012

Transmission diffraction gratings operating at 1,565 nm based on multilayer porous silicon films are modeled, fabricated, and tested. Features down to 2 μm have been patterned into submicron-thick mesoporous films using standard photolithographic and dry etching techniques. After patterning of the top porous film, a second anodization can be performed, allowing an under-layer of highly uniform porosity and thickness to be achieved. High transmission greater than 40% is measured, and modeling results suggest that a change in diffraction efficiency of 1 dB for a 1% change in normalized refractive index can be achieved. Preliminary measurement of solvent vapor shows a large signal change from the grating sensor in agreement with models.
A critical stress model for cell motility
Authors: Mehrayin Mehrnush , Farmanzad Farhad , Mozafari Masoud , Vashaee Daryoosh , Tayebi Lobat
Published: November 2012

A detailed theoretical model that combines the conventional viscoelastic continuum description of cell motion with a dynamic active stress is presented. The model describes the ameboid cells movement comprising of protrusion and adhesion of the front edge followed by detachment and movement of the tail. Unlike the previous viscoelastic descriptions in which the cell movement is steady, the presented model describes the “walking” of the cell in response to specific active stress components acting separately on the front and rear of the cell. In this locomotive model first the tail of the cell is attached to the substrate and active stress is applied to the front of the cell. Consequently, the stress in the tail increases. When the stress in the tail exceeds a critical value, namely critical stress, the conditions are updated so that the front is fixed and the tail of the cell is detached from the substrate and moves towards the front. Consequently, the stress in the tail decreases. When the stress goes to zero, the starting conditions become active and the process continues. At start the cell is stretched and its length is increased as the front of cell migrates more than the rear. However, after several steps the front and rear move equally and the cell length stays constant during the movement. In this manuscript we analyzed such cell dynamics including the length variation and moving velocity. Finally, by considering this fact that at the single-cell level, interactions with the extracellular environment occur on a nanometer length scale, the value of critical stress was estimated.
Factors associated with home delivery in Bahirdar, Ethiopia: A case control study
Authors: Abebe Fantu , Berhane Yemane , Girma Belaineh
Published: November 2012

Background

In Ethiopia although pregnant mothers increasingly attend antenatal clinics, utilization of skilled delivery service remains very low. The individual or health system factors that affect women’s preferences for delivery places are not well known.

Method

A case control study was conducted in July 2010 to assess factors associated with utilization of institutional delivery service. A total of 324 mothers who recently delivered and visited either postnatal care or sought immunization services were included. Cases (n = 108) were mothers who gave birth at home and controls (n = 216) were those who delivered at health facility. Pre-tested and standardized questionnaires were used to collect relevant data by trained data collectors. Logistic regression model was used to control for confounding.

Result

The likelihood of delivering at home was greater among mothers with inadequate knowledge of pregnancy related services (AOR = 62, 95% CI: 3, 128.4), those who started attending ANC after 24 weeks of gestation (AOR 8.7, 95% CI: 2.2, 33.3), mothers having no formal education (Adjusted OR 4.2, 95% CI 1.63, 11.27) and rural residents (AOR = 3.6, 95%CI: 1.4, 9.0).

Conclusion

The predominant factors associated with home delivery services were lack of knowledge about obstetrics care, delay in starting Antenatal Care (ANC) follow up, having, Illiteracy and rural residence. Audience specific behavioral change communication should be designed to improve the demand for delivery services. Health professionals should take the opportunity to encourage mothers attend delivery services during ANC follow up. Improvements should be made in social conditions including literacy and major social mobilization endeavors.
Morphological and microsatellite DNA diversity of Nigerian indigenous sheep
Authors: Agaviezor Brilliant O , Peters Sunday O , Adefenwa Mufliat A , Yakubu Abdulmojeed , Adebambo Olufunmilayo A , Ozoje Michael O , Ikeobi Christian ON , Wheto Matthew , Ajayi Oyeyemi O , Amusan Samuel A , Ekundayo Oludotun J , Sanni Timothy M , Okpeku Moses , Onasanya Gbolabo O , De Donato Marcos , Ilori Babatunde M , Kizilkaya Kadir , Imumorin Ikhide G
Published: November 2012

Background

Sheep is important in the socio-economic lives of people around the world. It is estimated that more than half of our once common livestock breeds are now endangered. Since genetic characterization of Nigerian sheep is still lacking, we analyzed ten morphological traits on 402 animals and 15 microsatellite DNA markers in 384 animals of the 4 Nigerian sheep breeds to better understand genetic diversity for breeding management and germplasm conservation.

Results

Morphological traits of Uda and Balami were significantly (P < 0.05) higher than Yankasa, which were both higher than West African Dwarf (WAD) sheep. Stepwise discriminant analysis showed tail length, rump height, chest girth, ear length and chest depth as the most discriminating variables for classification. Mahalanobis distances show the least differentiation between Uda and Balami and the largest between WAD and Balami sheep. While 93.3% of WAD sheep were correctly assigned to their source genetic group, 63.9% of Yankasa, 61.2% of Balami and 45.2% of Uda were classified correctly by nearest neighbour discriminant analysis. The overall high Polymorphism Information Content (PIC) of all microsatellite markers ranged from 0.751 to 0.927 supporting their use in genetic characterization. Expected heterozygosity was high for all loci (0.783 to 0.93). Mean heterozygote deficiency across all populations (0.171 to 0.534) possibly indicate significant inbreeding (P < 0.05). Mean values for F

ST
, F
IT
and F
IS
statistics across all loci were 0.088, 0.394 and 0.336 respectively. Yankasa and Balami are the most closely related breeds (DA = 0.184) while WAD and Balami are the farthest apart breeds (DA = 0.665), which is coincident with distance based on morphological analysis and population structure assessed by STRUCTURE.

Conclusions

These results suggest that within-breed genetic variation in Nigerian sheep is higher than between-breeds and may be a valuable tool for genetic improvement and conservation. The higher genetic variability in Yankasa suggests the presence of unique ancestral alleles reflecting the presence of certain functional genes which may result in better adaptability in more agro-ecological zones of Nigeria. These genetic characteristics are potentially useful in planning improvement and conservation strategies in Nigerian indigenous sheep.
An indoor augmented-reality evacuation system for the Smartphone using personalized Pedometry
Authors: Ahn Junho , Han Richard
Published: November 2012

There currently exist widely used mobile phone emergency applications for the smartphone and limited mobile emergency applications for indoor environments. However, the outdoor applications only focus primarily on providing accident information to users, and the indoor applications are limited by the unavailability of GPS user-positioning and by WiFi-based access problems. To compensate for these limitations, we propose the RescueMe system, which uses an indoor mobile Augmented Reality application, personalized pedometry, and an optimal exit path algorithm. Together these components comprise a system that can quickly and easily recommend an efficient exit path to mobile phone users in emergency situations. We have developed the mobile-based RescueMe system for use in large-scale buildings that contain complex paths. We show how RescueMe leverages the sensors on a smartphone and utilizes Augmented Reality, cloud information, daily-based user walking patterns, and an adaptive GPS connection method, to deliver critical evacuation information to mobile phone users in indoor emergency situations.
Simultaneous production of animal feed enzymes (endoxylanase and endoglucanase) by Penicillium janthinellum from waste jute caddies
Authors: Kundu Aditi , Karmakar Moumita , Ray Rina Rani
Published: November 2012

Background

In West Bengal, India, a huge amount of lignocellulosic jute waste products, in the form of unutilized cut pieces of jute caddies consisting 65% cellulose, 22.5% hemicellulose, and 11% lignin, is generated from jute industries. Naturally, the high percentage of lignocellulolytic material made it an attractive substrate for the hydrolytic cleavage of lignocellulolytic enzymes, namely xylanases and cellulases. The present study aims for the production of xylanolytic and cellulolytic enzymes by a fungal isolate Penicillium janthinellum MTCC10889 using a cheap medium containing residual jute caddies obtained from jute mill wastes and chemically defined basal medium under submerged fermentation.

Results

The fungal isolate P. janthinellum MTCC10889 is an outstanding producer of endoxylanase and also cellulase. A central composite design was applied to optimize its lignocellulolytic as well as cellulolytic enzyme production in submerged-state fermentation. The impact of three quantitative variables, namely pH, temperature, and substrate concentrations, on enzyme production was investigated by using a chemically defined basal medium supplemented with jute caddy as substrate. Such optimum conditions were as follows: pH 7.47, temperature of 30.35°C, and substrate concentration of 1.70% for endoxylanase, and pH 7.51, temperature of 29.69°C, and substrate concentration of 2.11% for cellulase production.

Conclusions

A combination of one-factor-at-a-time approach is replaced by response surface methodology using central composite design, which resulted in 3.08- and 3.84-fold increases in the yields of endoxylanase (1,750 IU/ml) and endoglucanase (192 IU/ml), respectively. The highest endoxylanase and endoglucanase productions under optimized conditions were achieved within 48 h of growth.
Functional and histopathological identification of the respiratory failure in a DMSXL transgenic mouse model of Myotonic Dystrophy
Authors: Panaite Petrica-Adrian , Kuntzer Thierry , Gourdon Geneviève , Lobrinus Johannes Alexander , Barakat-Walter Ibtissam
Published: November 2012

Acute and chronic respiratory failure is one of the major and potentially life-threatening features in patients with Myotonic Dystrophy type 1 (DM1). Despite the several clinical demonstrations showing respiratory problems in DM1 patients, the mechanisms are still not completely known. This study was designed to investigate whether the DMSXL transgenic mouse model for DM1 exhibit respiratory disorder, and if so to identify the pathological changes underlying these respiratory problems. Using pressure plethysmography we assessed the breathing function in control mice and DMSXL mice generated after large expansions of CTG repeat in successive generations of DM1 transgenic mice. The statistical analysis of breathing function measurements revealed a significant decrease in the most relevant respiratory parameters in DMSXL mice, indicating impaired respiratory function. The histological and morphometric analysis showed pathological changes in diaphragmatic muscle of DMSXL mice characterized by an increase in the percentage of type I muscle fibers, the presence of central nuclei, partial denervation of end-plates (EPs) and a significant reduction in their size, shape complexity and density of acetylcholine receptors, all of which reflect a possible break down in communication between the diaphragmatic muscles fibers and the nerve terminals. Diaphragm muscle abnormalities were accompanied by an accumulation of mutant DMPK RNA foci in muscle fiber nuclei. Moreover, in DMSXL mice, the unmyelinated phrenic afferents are significantly lower. Also in these mice, significant neuronopathy was not detected in either cervical phrenic motor neurons or brainstem respiratory neurons. Since EPs are involved in the transmission of action potentials and the unmyelinated phrenic afferents exert a modulating influence on the respiratory drive, the pathological alterations affecting these structures may underlie the respiratory impairment detected in DMSXL mice. Understanding mechanisms of respiratory deficiency should guide pharmaceutical and clinical research towards better therapy for the respiratory deficits associated with DM1.
Myosin isoform switching during assembly of the Drosophila flight muscle thick filament lattice
Authors: Orfanos Zacharias , Sparrow John C.
Published: November 2012

During muscle development myosin molecules form symmetrical thick filaments, which integrate with the thin filaments to produce the regular sarcomeric lattice. In Drosophila indirect flight muscles (IFMs) the details of this process can be studied using genetic approaches. The weeP26 transgenic line has a GFP-encoding exon inserted into the single Drosophila muscle myosin heavy chain gene, Mhc. The weeP26 IFM sarcomeres have a unique MHC-GFP-labelling pattern restricted to the sarcomere core, explained by non-translation of the GFP exon following alternative splicing. Characterisation of wild-type IFM MHC mRNA confirmed the presence of an alternately spliced isoform, expressed earlier than the major IFM-specific isoform. The two wild-type IFM-specific MHC isoforms differ by the presence of a C-terminal ‘tailpiece’ in the minor isoform. The sequential expression and assembly of these two MHCs into developing thick filaments suggest a role for the tailpiece in initiating A-band formation. The restriction of the MHC-GFP sarcomeric pattern in weeP26 is lifted when the IFM lack the IFM-specific myosin binding protein flightin, suggesting that it limits myosin dissociation from thick filaments. Studies of flightin binding to developing thick filaments reveal a progressive binding at the growing thick filament tips and in a retrograde direction to earlier assembled, proximal filament regions. We propose that this flightin binding restricts myosin molecule incorporation/dissociation during thick filament assembly and explains the location of the early MHC isoform pattern in the IFM A-band.
HIV-1 Tat protein inhibits neurosecretion by binding to phosphatidylinositol 4,5-bisphosphate
Authors: Tryoen-Tóth Petra , Chasserot-Golaz Sylvette , Tu Annie , Gherib Patricia , Bader Marie-France , Beaumelle Bruno , Vitale Nicolas
Published: November 2012

HIV-1 transcriptional activator (Tat) enables viral transcription and is also actively released by infected cells. Extracellular Tat can enter uninfected cells and affect some cellular functions. Here, we examine the effects of Tat protein on the secretory activity of neuroendocrine cells. When added to the culture medium of chromaffin and PC12 cells, Tat was actively internalized and strongly impaired exocytosis as measured by carbon fiber amperometry and growth hormone release assay. Expression of Tat mutants that do not bind to phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] did not affect secretion, and overexpression of phosphatidylinositol 4-phosphate 5-kinase (PIP5K), the major PtdIns(4,5)P2 synthesizing enzyme, significantly rescued the Tat-induced inhibition of neurosecretion. This suggests that the inhibition of exocytosis may be the consequence of PtdIns(4,5)P2 sequestration. Accordingly, expression of Tat in PC12 cells interfered with the secretagogue-dependent recruitment of annexin A2 to the plasma membrane, a PtdIns(4,5)P2-binding protein that promotes the formation of lipid microdomains that are required for exocytosis. In addition Tat significantly prevented the reorganization of the actin cytoskeleton necessary for the movement of secretory vesicles towards plasma membrane fusion sites. Thus, the capacity of extracellular Tat to enter neuroendocrine cells and sequester plasma membrane PtdIns(4,5)P2 perturbs several PtdIns(4,5)P2-dependent players of the exocytotic machinery, thereby affecting neurosecretion. We propose that Tat-induced inhibition of exocytosis is involved in the neuronal disorders associated with HIV-1 infection.
An intracellular membrane junction consisting of flagellum adhesion glycoproteins links flagellum biogenesis to cell morphogenesis in Trypanosoma brucei
Authors: Sun Stella Y. , Wang Chao , Yuan Y. Adam , He Cynthia Y.
Published: November 2012

African trypanosomes have a single, membrane-bounded flagellum that is attached to the cell cortex by membrane adhesion proteins and an intracellular flagellum attachment zone (FAZ) complex. The coordinated assembly of flagellum and FAZ, during the cell cycle and the life cycle development, plays a pivotal role in organelle positioning, cell division and cell morphogenesis. To understand how the flagellum and FAZ assembly are coordinated, we examined the domain organization of the flagellum adhesion protein 1 (FLA1), a glycosylated, transmembrane protein essential for flagellum attachment and cell division. By immunoprecipitation of a FLA1-truncation mutant that mislocalized to the flagellum, a novel FLA1-binding protein (FLA1BP) was identified in procyclic Trypanosoma brucei. The interaction between FLA1 on the cell membrane and FLA1BP on the flagellum membrane acts like a molecular zipper, joining flagellum membrane to cell membrane and linking flagellum biogenesis to FAZ elongation. By coordinating flagellum and FAZ assembly during the cell cycle, morphology information is transmitted from the flagellum to the cell body.
β amyloid protein precursor-like (Appl) is a Ras1/MAPK-regulated gene required for axonal targeting in Drosophila photoreceptor neurons
Authors: Mora Natalia , Almudi Isabel , Alsina Berta , Corominas Montserrat , Serras Florenci
Published: November 2012

In a genome-wide expression profile search for genes required for Drosophila R7 photoreceptor development we found β amyloid protein precursor-like (Appl), the ortholog of human APP, which is a key factor in the pathogenesis of Alzheimer's disease. We analyzed Appl expression in the eye imaginal disc and found that is highly accumulated in R7 photoreceptor cells. The R7 photoreceptor is responsible for UV light detection. To explore the link between high expression of Appl and R7 function, we have analyzed Appl null mutants and found reduced preference for UV light, probably because of mistargeted R7 axons. Moreover, axon mistargeting and inappropriate light discrimination are enhanced in combination with neurotactin mutants. R7 differentiation is triggered by the inductive interaction between R8 and R7 precursors, which results in a burst of Ras1/MAPK, activated by the tyrosine kinase receptor Sevenless. Therefore, we examined whether Ras1/MAPK is responsible for the high Appl expression. Inhibition of Ras1 signaling leads to reduced Appl expression, whereas constitutive activation drives ectopic Appl expression. We show that Appl is directly regulated by the Ras/MAPK pathway through a mechanism mediated by PntP2, an ETS transcription factor that specifically binds ETS sites in the Appl regulatory region. We also found that zebrafish appb expression increased after ectopic fgfr activation in the neural tube of zebrafish embryos, suggesting a conserved regulatory mechanism.
Ubiquitin-independent function of optineurin in autophagic clearance of protein aggregates
Authors: Korac Jelena , Schaeffer Veronique , Kovacevic Igor , Clement Albrecht M. , Jungblut Benno , Behl Christian , Terzic Janos , Dikic Ivan
Published: November 2012

Aggregation of misfolded proteins and the associated loss of neurons are considered a hallmark of numerous neurodegenerative diseases. Optineurin is present in protein inclusions observed in various neurodegenerative diseases including amyotrophic lateral sclerosis (ALS), Huntington's disease, Alzheimer's disease, Parkinson's disease, Creutzfeld-Jacob disease and Pick's disease. Optineurin deletion mutations have also been described in ALS patients. However, the role of optineurin in mechanisms of protein aggregation remains unclear. In this report, we demonstrate that optineurin recognizes various protein aggregates via its C-terminal coiled-coil domain in a ubiquitin-independent manner. We also show that optineurin depletion significantly increases protein aggregation in HeLa cells and that morpholino-silencing of the optineurin ortholog in zebrafish causes the motor axonopathy phenotype similar to a zebrafish model of ALS. A more severe phenotype is observed when optineurin is depleted in zebrafish carrying ALS mutations. Furthermore, TANK1 binding kinase 1 (TBK1) is colocalized with optineurin on protein aggregates and is important in clearance of protein aggregates through the autophagy-lysosome pathway. TBK1 phosphorylates optineurin at serine 177 and regulates its ability to interact with autophagy modifiers. This study provides evidence for a ubiquitin-independent function of optineurin in autophagic clearance of protein aggregates as well as additional relevance for TBK1 as an upstream regulator of the autophagic pathway.
Specific roles of Gi protein family members revealed by dissecting SST5 coupling in human pituitary cells
Authors: Peverelli Erika , Busnelli Marta , Vitali Eleonora , Giardino Elena , Galés Celine , Lania Andrea G. , Beck-Peccoz Paolo , Chini Bice , Mantovani Giovanna , Spada Anna
Published: November 2012

Despite intensive investigation over the past 20 years, the specific role played by individual Gi protein family members in mediating complex cellular effects is still largely unclear. Therefore, we investigated the role of specific Gi proteins in mediating somatostatin (SS) effects in somatotroph cells. Because our previous data showed that SS receptor type 5 (SST5) carrying a spontaneous R240W mutation in the third intracellular loop had a similar ability to inhibit intracellular cAMP levels to the wild-type protein but failed to mediate inhibition of growth hormone (GH) release and cell proliferation, we used this model to check specific receptor–G-protein coupling by a bioluminescent resonance energy transfer analysis. In HEK293 cells, wild-type SST5 stimulated the activation of Gαi1–3 and GαoA, B, whereas R240W SST5 maintained the ability to activate Gαi1–3 and GαoB, but failed to activate the splicing variant GαoA. To investigate the role of the selective deficit in GαoA coupling, we co-transfected human adenomatous somatotrophs with SST5 and a pertussis toxin (PTX)-resistant GαoA (GαoA(PTX-r)) protein. In PTX-treated cells, GαoA(PTX-r) rescued the ability of the selective SST5 analog BIM23206 to inhibit extracellular signal-related kinase 1/2 (ERK1/2) phosphorylation, GH secretion and intracellular cAMP levels. Moreover, we demonstrated that silencing of GαoA completely abolished SST5-mediated inhibitory effects on GH secretion and ERK1/2 phosphorylation, but not on cAMP levels. In conclusion, by analysing the coupling specificity of human SST5 to individual Gαi and Gαo subunits, we identified a crucial role for GαoA signalling in human pituitary cells.
Activation of Notch in lgd mutant cells requires the fusion of late endosomes with the lysosome
Authors: Schneider Markus , Troost Tobias , Grawe Ferdi , Martinez-Arias Alfonso , Klein Thomas
Published: November 2012

The tumour suppressor Lethal (2) giant discs (Lgd) is a regulator of endosomal trafficking of the Notch signalling receptor as well as other transmembrane proteins in Drosophila. The loss of its function results in an uncontrolled ligand-independent activation of the Notch signalling receptor. Here, we investigated the consequences of loss of lgd function and the requirements for the activation of Notch. We show that the activation of Notch in lgd cells is independent of Kuz and dependent on γ-secretase. We found that the lgd cells have a defect that delays degradation of transmembrane proteins, which are residents of the plasma membrane. Furthermore, our results show that the activation of Notch in lgd cells occurs in the lysosome. By contrast, the pathway is activated at an earlier phase in mutants of the gene that encodes the ESCRT-III component Shrub, which is an interaction partner of Lgd. We further show that activation of Notch appears to be a general consequence of loss of lgd function. In addition, electron microscopy of lgd cells revealed that they contain enlarged multi-vesicular bodies. The presented results further elucidate the mechanism of uncontrolled Notch activation upon derailed endocytosis.
Protein tyrosine phosphatase SHP2 suppresses podosome rosette formation in Src-transformed fibroblasts
Authors: Pan Yi-Ru , Cho Ke-Huan , Lee Hsiao-Hui , Chang Zee-Fen , Chen Hong-Chen
Published: November 2012

Podosomes are actin-enriched membrane protrusions that play important roles in extracellular matrix degradation and invasive cell motility. Podosomes undergo self-assembly into large rosette-like structures in Src-transformed fibroblasts, osteoclasts and certain highly invasive cancer cells. Several protein tyrosine kinases have been shown to be important for the formation of podosome rosettes, but little is known regarding the role of protein tyrosine phosphatases in this process. We found that knockdown of the Src homolog domain-containing phosphatase 2 (SHP2) significantly increased podosome rosette formation in Src-transformed fibroblasts. By contrast, SHP2 overexpression suppressed podosome rosette formation in these cells. The phosphatase activity of SHP2 was essential for the suppression of podosome rosette formation. SHP2 selectively suppressed the tyrosine phosphorylation of Tks5, a scaffolding protein required for podosome formation. The inhibitory effect of SHP2 on podosome rosette formation was associated with the increased activation of Rho-associated kinase (ROCK) and the enhanced polymerization of vimentin filaments. A higher content of polymerized vimentin filaments was correlated with a lower content of podosome rosettes. Taken together, our findings indicate that SHP2 serves as a negative regulator of podosome rosette formation through the dephosphorylation of Tks5 and the activation of ROCK-mediated polymerization of vimentin in Src-transformed fibroblasts.
The endocannabinoid pathway and the female reproductive organs
Authors: Di Blasio Anna Maria , Vignali Michele , Gentilini Davide
Published: November 2012

Endocannabinoids are endogenous ligands of cannabinoid, vanilloid and peroxisome proliferator-activated receptors that activate multiple signal transduction pathways. Together with their receptor and the enzymes responsible for their synthesis and degradation, these compounds constitute the endocannabinoid system that has been recently shown to play, in humans, an important role in modulating several central and peripheral functions including reproduction. Given the relevance of the system, drugs that are able to interfere with the activity of endocannabinoids are currently considered as candidates for the treatment of various diseases. In this review, we will summarise the current knowledge regarding the effects of endocannabinoids in female reproductive organs. In particular, we will focus on some newly reported mechanisms that can affect endometrial plasticity both in physiological and in pathological conditions.
Hormonal regulation of β-myosin heavy chain expression in the mouse left ventricle
Authors: Patrizio Mario , Musumeci Marco , Piccone Ambra , Raggi Carla , Mattei Elisabetta , Marano Giuseppe
Published: November 2012

We investigated the influence of sex hormones on the expression of α- and β-cardiac myosin heavy chain isoforms (α-MHC and β-MHC) in C57bl/6 mice of both sexes under physiological and pathological conditions. In the left ventricles (LVs) of fertile female mice, β-MHC expression was tenfold higher compared with the age-matched males, whereas no difference was found in α-MHC expression. These differences disappeared after ovariectomy or in immature mice. We also found a sex-related difference in expression of β-adrenoceptors (β1-AR), as mRNA levels of this gene were 40% lower in fertile females compared with males of the same age but did not differ in prepubertal or ovariectomized animals. Interestingly, the deletion of both β1- and β2-ARs abolished sex difference of β-MHC expression, as mRNA levels in the LVs of knockout males were increased and reached values comparable to those of knockout females. Moreover, the β1-AR antagonist metoprolol induced about a threefold increase in β-MHC expression in adult male mice. The capability of gender to regulate β-MHC expression was also evaluated in the presence of hemodynamic overload. Thoracic aortic coarctation (TAC) produced cardiac hypertrophy along with a 12-fold increase in β-MHC and a 50% decrease in β1-AR expression in males but not in females, thus abolishing the gender difference observed in sham animals for such genes. By contrast, TAC did not change β2-AR expression. In conclusion, our results show that the expression of β-MHC and β1-AR in the LVs undergo gender-related and correlated changes under both physiological and pathological conditions and suggest a role of β1-AR-mediated signaling.
Mitotane induces CYP3A4 expression via activation of the steroid and xenobiotic receptor
Authors: Takeshita Akira , Igarashi-Migitaka Junko , Koibuchi Noriyuki , Takeuchi Yasuhiro
Published: November 2012

Adrenocortical carcinoma (ACC) is a rare disease with an extremely poor prognosis. Mitotane alone or in combination with other cytotoxic drugs is a common therapeutic option for ACC. In addition to its adrenolytic function, mitotane has been known for decades to increase the metabolic clearance of glucocorticoids. It was recently shown that the tyrosine kinase inhibitor sunitinib is also rapidly metabolized in patients treated with mitotane, indicating that mitotane engages in clinically relevant drug interactions. Although the precise mechanism of these interactions is not well understood, cytochrome P450 mono-oxygenase 3A4 (CYP3A4) is a key enzyme to inactivate both glucocorticoids and sunitinib. The nuclear receptor steroid and xenobiotic receptor (SXR (NR1I2)) is one of the key transcriptional regulators ofCYP3A4gene expression in the liver and intestine. A variety of xenobiotics bind to SXR and stimulate transcription of xenobiotic-response elements (XREs) located in theCYP3A4gene promoter. In this study, we evaluated the effects of mitotane on SXR-mediated transcriptionin vitroby luciferase reporter analysis, SXR–steroid receptor coactivator 1 (SRC1) interactions, quantitative real-time PCR analysis ofCYP3A4expression, SXR knockdown, and CYP3A4 enzyme activity assays using human hepatocyte-derived cells. We found that mitotane activated SXR-mediated transcription of the XREs. Mitotane recruited SRC1 to the ligand-binding domain of SXR. Mitotane increasedCYP3A4mRNA levels, which was attenuated by SXR knockdown. Finally, we showed that mitotane increased CYP3A4 enzyme activity. We conclude that mitotane can induceCYP3A4gene expression and suggest that mitotane is used cautiously due to its drug–drug interactions.
Proteomic analysis of human cervicovaginal fluid collected before preterm premature rupture of the fetal membranes
Authors: Liong Stella , Di Quinzio Megan K W , Heng Yujing J , Fleming Gabrielle , Permezel Michael , Rice Gregory E , Georgiou Harry M
Published: November 2012

A significant obstetric complication facing contemporary materno–fetal medicine is preterm premature rupture of the fetal membranes (preterm PROM), which occurs in 30% of all preterm births. The objective of this study was to identify differentially expressed proteins in the cervicovaginal fluid of asymptomatic women before the clinical manifestation of preterm PROM. The preterm PROM group comprised of women with samples collected 6–23 days before PROM, who subsequently delivered preterm (n=5). Women who spontaneously delivered at term served as gestation-matched controls (n=10). Two-dimensional difference in-gel electrophoresis was used to distinguish differential expression between the pooled groups and fold changes were subsequently confirmed by two-dimensional PAGE of individual samples. Spots of interest were identified by mass spectrometry. Proteins that were significantly reduced with impending preterm PROM included the following: thioredoxin (2.7-fold), interleukin 1 receptor antagonist (1.7-fold), fatty acid-binding protein 5 (2.1-fold), cystatin A (dimer; 1.9-fold), monocyte/neutrophil elastase inhibitor (1.6-fold), squamous cell carcinoma antigen-1 (2.1-fold) and γ-glutamyl cyclotransferase (3.0-fold). By contrast, annexin A3 (3.7-fold) and vitamin D binding protein (3.9-fold) were significantly increased with impending preterm PROM. Western blot analysis was also performed on an independent cohort of preterm PROM and control samples to validate these candidate biomarkers. These proteins have known biological functions in oxidative balance, anti-inflammatory activity, metabolism or protease inhibition that may facilitate membrane rupture.
Morfofisiologia e anatomia foliar de mudas micropropagadas e aclimatizadas de abacaxizeiro cv. Smooth Cayenne em diferentes substratos
Authors: Silva Adriano Bortolotti da , Pasqual Moacir , Araujo Aparecida Gomes de , Braga Francyane Tavares , Castro Evaristo Mauro de , Albert Lúcia Helena de Barros
Published: November 2012

Objetivou-se com este trabalho determinar o melhor substrato e o tempo ideal de aclimatização em plantas de abacaxizeiro cv. Smooth Cayenne e comparar anatomicamente plântulas in vitro e aclimatizadas. Gemas foram inoculadas em meio MS + 6,66 µM de BAP e, após três subcultivos, as brotações foram transferidas para meio MS, por 30 dias. Os meios foram solidificados com 0,7% de ágar e tiveram seu pH ajustado para 5,8 antes da autoclavagem (120 ºC por 20 minutos). A incubação foi em sala de crescimento (25 ± 1 ºC, irradiância de 35 μmol m-2 s-1 e fotoperíodo de 16 h). Os brotos obtidos in vitro foram transplantados para tubetes, em casa de vegetação, contendo os seguintes substratos: T1- Plantmax® + húmus, T2- Plantmax® + vermiculita e T3- Plantmax®. Cortes anatômicos foram efetuados em folhas de propágulos mantidos in vitro e em plantas com 20, 40 e 60 dias de aclimatização nos diferentes substratos. A utilização do substrato Plantmax® + húmus na aclimatização propiciou maior desenvolvimento das mudas micropropagadas. A partir de 40 dias da aclimatização, as plantas apresentaram características anatômicas que podem favorecer a sua adaptação às condições de campo.
Glucosamine and chondroitin sulfate in the repair of osteochondral defects in dogs - clinical-radiographic analysis
Authors: Eleotério Renato Barros , Borges Andréa Pacheco Batista , Pontes Kelly Cristine de Sousa , Fernandes Natália Alves , Soares Priscila Ferreira , Silva Mariana Brettas , Martins Naira Jandafet Sampaio , Machado João Paulo
Published: November 2012

Among the proposed treatments to repair lesions of degenerative joint disease (DJD), chondroprotective nutraceuticals composed by glucosamine and chondroitin sulfate are a non-invasive theraphy with properties that favors the health of the cartilage. Although used in human, it is also available for veterinary use with administration in the form of nutritional supplement independent of prescription, since they have registry only in the Inspection Service, which does not require safety and efficacy testing. The lack of such tests to prove efficacy and safety of veterinary medicines required by the Ministry of Agriculture and the lack of scientific studies proving its benefits raises doubts about the efficiency of the concentrations of such active substances. In this context, the objective of this study was to evaluate the efficacy of a veterinary chondroprotective nutraceutical based on chondroitin sulfate and glucosamine in the repair of osteochondral defects in lateral femoral condyle of 48 dogs, through clinical and radiographic analysis. The animals were divided into treatment group (TG) and control group (CG), so that only the TG received the nutraceutical every 24 hours at the rate recommended by the manufacturer. The results of the four treatment times (15, 30, 60 and 90 days) showed that the chondroprotective nutraceutical, in the rate, formulation and administration at the times used, did not improve clinical signs and radiologically did not influence in the repair process of the defects, since the treated and control groups showed similar radiographic findings at the end of the treatments.