bluuebunny/crossref_metadata_2025_split huggingface dataset:
The proteome of exudates from germinating Lupinus albus seeds is secreted through a selective dual‐step process and contains proteins involved in plant defence
Authors: Scarafoni Alessio , Ronchi Alessandro , Prinsi Bhakti , Espen Luca , Assante Gemma , Venturini Giovanni , Duranti Marcello 
Published: January 2013

The general knowledge of defence activity during the first steps of seed germination is still largely incomplete. The present study focused on the proteins released in the exudates of germinating white lupin seeds. During the first 24 h, a release of proteins was observed. Initially (i.e. during the first 12 h), the proteins found in exudates reflected the composition of the seed, indicating a passive extrusion of pre‐formed proteins. Subsequently, when the rate of protein release was at its highest, the composition of the released proteome changed drastically. This transition occurred in a short time, indicating that more selective and regulated events, such as secretory processes, took place soon after the onset of germination. The present study considered: (a) the characterization of the proteome accumulated in the germinating medium collected after the appearance of the post‐extrusion events; (b) the biosynthetic origin and the modalities that are the basis of protein release outside the seeds; and (c) an assessment of antifungal activity of these exudates. The most represented protein in the exudate was chitinase, which was synthesized de novo. The other proteins are involved in the cellular mechanisms responding to stress events, including biotic ones. This exudate was effectively able to inhibit fungal growth. The results of the present study indicate that seed exudation is a dual‐step process that leads to the secretion of selected proteins and thus is not a result of passive leakage. The released proteome is involved in protecting the spermosphere environment and thus may act as first defence against pathogens.
Aromatic substitution of the FAD‐shielding tryptophan reveals its differential role in regulating electron flux in methionine synthase reductase and cytochrome P450 reductase
Authors: Meints Carla E. , Simtchouk Svetlana , Wolthers Kirsten R. 
Published: January 2013

Methionine synthase reductase (MSR) and cytochrome P450 reductase (CPR) transfer reducing equivalents from NADPH via an FAD and FMN cofactor to a redox partner protein. In both enzymes, hydride transfer from NADPH to FAD requires displacement of a conserved tryptophan that lies coplanar to the FAD isoalloxazine ring. Swapping the tryptophan for a smaller aromatic side chain revealed a distinct role for the residue in regulating MSR and CPR catalysis. MSR W697F and W697Y showed enhanced catalysis, noted by increases in kcat and kcat/Km(NADPH) for steady‐state cytochrome c3+ reduction and a 10‐fold increase in the rate constant (kobs1) associated with hydride transfer. Elevated primary kinetic isotope effects on kobs1 for W697F and W697Y suggest that preceding isotopically insensitive steps like displacement of W697 are less rate determining. MSR W697Y, but not MSR W697F, showed detectable formation of the disemiquinone intermediate, indicating that the polarity of the aromatic side chain influences the rate of interflavin electron transfer. By contrast, the CPR variants (W676F and W676Y) displayed modest decreases in cytochrome c3+ reduction, a 30‐ and 3.5‐fold decrease in the rate of FAD reduction, accumulation of a FADH2–NADP+ charge‐transfer complex and dramatically suppressed rates of interflavin electron transfer. We conclude for MSR that hydride transfer is ‘gated’ by the free energy required to disrupt dispersion forces between the FAD isoalloxazine ring and W697. By contrast, the bulky indole ring of W676 accelerates catalysis in CPR by lowering the energy barrier for displacement of the oxidized nicotinamide ring coplanar with the FAD.
Recombinant purine nucleoside phosphorylases from thermophiles: preparation, properties and activity towards purine and pyrimidine nucleosides
Authors: Zhou Xinrui , Szeker Kathleen , Janocha Bernd , Böhme Thomas , Albrecht Dirk , Mikhailopulo Igor A. , Neubauer Peter 
Published: January 2013

Thermostable nucleoside phosphorylases are attractive biocatalysts for the synthesis of modified nucleosides. Hence we report on the recombinant expression of three ‘high molecular mass’ purine nucleoside phosphorylases (PNPs) derived from the thermophilic bacteria Deinococcus geothermalis, Geobacillus thermoglucosidasius and from the hyperthermophilic archaeon Aeropyrum pernix (5′–methythioadenosine phosphorylase; ApMTAP). Thermostability studies, kinetic analysis and substrate specificities are reported. The PNPs were stable at their optimal temperatures (DgPNP, 55 °C; GtPNP, 70 °C; ApMTAP, activity rising to 99 °C). Substrate properties were investigated for natural purine nucleosides [adenosine, inosine and their C2′‐deoxy counterparts (activity within 50–500 U·mg−1)], analogues with 2′‐amino modified 2′‐deoxy‐adenosine and ‐inosine (within 0.1–3 U·mg−1) as well as 2′‐deoxy‐2′‐fluoroadenosine (9) and its C2′‐arabino diastereomer (10, within 0.01–0.03 U·mg−1). Our results reveal that the structure of the heterocyclic base (e.g. adenine or hypoxanthine) can play a critical role in the phosphorolysis reaction. The implications of this finding may be helpful for reaction mechanism studies or optimization of reaction conditions. Unexpectedly, the diastereomeric 2′‐deoxyfluoro adenine ribo‐ and arabino‐nucleosides displayed similar substrate properties. Moreover, cytidine and 2′‐deoxycytidine were found to be moderate substrates of the prepared PNPs, with substrate activities in a range similar to those determined for 2′‐deoxyfluoro adenine nucleosides 9 and 10. C2′‐modified nucleosides are accepted as substrates by all recombinant enzymes studied, making these enzymes promising biocatalysts for the synthesis of modified nucleosides. Indeed, the prepared PNPs performed well in preliminary transglycosylation reactions resulting in the synthesis of 2′‐deoxyfluoro adenine ribo‐ and arabino‐ nucleosides in moderate yield (24%).
Oral health of dental assistants and patients receiving maintenance – an investigation based on a district of Thuringia, Germany
Authors: Ziebolz D , Klopfleisch S , Fresmann S , Hornecker E , Mausberg RF 
Published: January 2013

AbstractObjectivesThe aim of the study was to assess data for the oral health behaviour and oral health of dental assistants (DAs) and patients (PTs) who regularly received preventive dental care.MethodsOne‐hundred DAs (38.8 years) and 100 PTs (44 years) participated in the study conducted in the Unstrut‐Hainich region of Thuringia, Germany. A questionnaire established oral hygiene and smoking habits. The dental examination included the caries index DMF‐T, evaluation of the periodontal situation (PSI) as well as gingival inflammation [papilla bleeding index (PBI)].ResultsSeventy‐seven DAs and 46 PTs used dental floss regularly. Twenty‐four DAs and 27 PTs were smokers. The mean DMF‐T of DAs was 12.5, which was significantly lower than the DMF‐T of PTs (17.0) (P < 0.001). The main reason for this difference was the number of missing teeth (DAs: 0.9, PTs: 4.8; P < 0.001). In both groups, several participants required periodontal treatment. Nevertheless, the number of DAs with PSI scores 3 or 4 was significantly lower (9%) than the corresponding number of PTs (63%) (P < 0.001). The PBI showed a significant difference between the groups (DAs = 0.1, PTs = 0.3; P < 0.01).ConclusionsAlthough the oral health of the participating DAs clearly exceeded the results found for PTs, it was concluded that DAs did not avail themselves of their job‐related knowledge and skills and so failed to optimize their personal oral health behaviour.
Prevalence of and risk factors for isolation of meticillin‐resistant Staphylococcus spp. from dogs with pyoderma in northern California, USA
Authors: Eckholm Nicole G. , Outerbridge Catherine A. , White Stephen D. , Sykes Jane E. 
Published: January 2013

Background –  Canine pyodermas associated with meticillin‐resistant Staphylococcus spp. (MRS) have increased in prevalence over the past decade.Hypothesis/Objectives –  To compare the prevalence of MRS isolation from dogs with superficial pyoderma at a primary care clinic (PCC) and those at a tertiary care facility (VMTH) in California, USA, and identify associated risk factors.Animals –  Client‐owned dogs from the VMTH (80 dogs) and the PCC (30 dogs).Methods –  Aerobic bacterial culture and antibiotic susceptibility were performed on swab specimens collected from dogs, and meticillin resistance was determined using microdilution methods according to Clinical and Laboratory Standards Institute guidelines. A mecA gene PCR assay was used to confirm meticillin resistance when possible.Results –  Of 89 staphylococcal isolates from the VMTH, 34 (38.2%) were meticillin resistant. In 31 dogs, pyoderma persisted, and one or more follow‐up isolates were obtained. The species isolated and drug susceptibility changed unpredictably during treatment. Of 33 PCC isolates, nine (27.3%) were meticillin resistant. Multiple drug resistance was identified in 41 of 53 (77.3%) MRS isolates from the VMTH and five of nine from the PCC. The sensitivity and specificity of PCR for the detection of meticillin resistance was 34 of 39 (87%) and 86 of 87 (99%), respectively. Risk factors for meticillin resistance for both sites were antibiotic treatment within the last year (P = 0.001), and for VMTH, hospitalization of dogs within the last year (P = 0.001).Conclusions and clinical importance –  The prevalence of meticillin resistance was not different between VMTH and PCC isolates (P = 0.29). Previous antimicrobial therapy was an important risk factor for the isolation of MRS at both sites.
The effect of ketoconazole on whole blood and skin ciclosporin concentrations in dogs
Authors: Gray Laura L. , Hillier Andrew , Cole Lynette K. , Rajala‐Schultz Päivi J. 
Published: January 2013

Background – Ciclosporin (CSA) is approved for the treatment of canine atopic dermatitis. Ciclosporin is metabolized by liver cytochrome P450 enzymes, a process inhibited by ketoconazole (KTZ).Hypothesis/Objectives – The aims of this study were to determine skin and blood CSA concentrations when CSA was administered alone at 5.0 (Treatment 1) or 2.5 mg/kg (Treatment 2) and when CSA was administered at 2.5 mg/kg concurrently with KTZ at 5 (Treatment 3) or 2.5 mg/kg (Treatment 4). We hypothesized that skin and blood CSA concentrations in Treatment 1 would not differ from those obtained with T3 or T4.Animals – In a randomized cross‐over study, six healthy research dogs received each of the treatments (Treatment 1, 2, 3 and 4) once daily for 7 days.Methods – After the first, fourth and seventh dose for each treatment, a peak and trough skin punch biopsy sample and whole blood sample were collected and analysed with high‐performance liquid chromatography–tandem mass spectrometry. Data were analysed using a repeated measures approach with PROC MIXED in SAS. Pairwise comparisons were performed with least squares means and Tukey–Kramer adjustment for multiple comparisons.Results – Mean blood CSA concentrations in Treatment 1 were not different from those in Treatment 2 or 4, but were less than in Treatment 3. Mean skin CSA concentrations in Treatment 1 were greater than in Treatment 2, not different from those in Treatment 4, and less than those in Treatment 3.Conclusions and clinical importance – Administration of CSA and KTZ concurrently at 2.5 mg/kg each may be as effective as CSA alone at 5.0 mg/kg for treatment of canine atopic dermatitis.
Fixing the skin barrier: past, present and future – man and dog compared
Authors: Marsella Rosanna 
Published: January 2013

Skin barrier dysfunction exists in both human and canine atopic dermatitis, leading to increased water loss and potentially facilitating allergen penetration and sensitization. Both lipid (e.g. ceramides) and protein (e.g. filaggrin) abnormalities have been described. Some are genetically inherited (e.g. filaggrin mutations are one of the major risk factors in humans) and some are secondary and linked to inflammation.In humans, numerous studies have shown efficacy of emollients and moisturizers in barrier restoration, and this approach has been for years the mainstay of therapy. Recently, this strategy has shown promise as a preventative function.In veterinary medicine, evidence regarding skin barrier impairment is rapidly building. Decreased ceramides and filaggrin (in some subsets of dogs) have been described. Altered metabolism of ceramides has also been proposed. Despite these preliminary data and the availability of products marketed to improve the skin barrier, evidence regarding the clinical benefit of skin repair intervention is still limited. Preliminary studies have demonstrated that topical application of fatty acids and ceramides and systemic administration of fatty acids improve lipid deficiencies in the skin of dogs with atopic dermatitis, but limited clinical evidence exists.Disease remission in humans is paralleled by an improved skin barrier, both with calcineurin inhibitors and glucocorticoids. In veterinary medicine, a preliminary study on ciclosporin and prednisone failed to detect significant improvement of water loss, while successful immunotherapy correlated with an improved skin barrier. Controlled, large studies are needed to address the question of which skin repair approach is clinically most effective and whether this can be used as a preventative strategy.
The canine and feline skin microbiome in health and disease
Authors: Weese J. Scott 
Published: January 2013

The skin harbours a diverse and abundant, yet inadequately investigated, microbial population. The population is believed to play an important role in both the pathophysiology and the prevention of disease, through a variety of poorly explored mechanisms.Early studies of the skin microbiota in dogs and cats reported a minimally diverse microbial composition of low overall abundance, most probably as a reflection of the limitations of testing methodology. Despite these limitations, it was clear that the bacterial population of the skin plays an important role in disease and in changes in response to both infectious and noninfectious diseases.Recent advances in technology are challenging some previous assumptions about the canine and feline skin microbiota and, with preliminary application of next‐generation sequenced‐based methods, it is apparent that the diversity and complexity of the canine skin microbiome has been greatly underestimated. A better understanding of this complex microbial population is critical for elucidation of the pathophysiology of various dermatological (and perhaps systemic) diseases and to develop novel ways to manipulate this microbial population to prevent or treat disease.
Epidemiology of human atopic dermatitis – seven areas of notable progress and seven areas of notable ignorance
Authors: Williams Hywel C. 
Published: January 2013

Background –  This narrative review highlights areas within the epidemiology of human atopic dermatitis (AD) where significant progress has been made and where considerable ignorance still exists. The review is supported by systematic reviews wherever possible, with the purpose of stimulating fresh approaches to human and veterinary research into AD.Progress –  Areas of progress include valid and repeatable methods of disease definition, global documentation of disease prevalence and impact, clarification of the role of some genetic factors, such as filaggrin gene mutations, clear evidence that environmental factors are key, as demonstrated by the positive social class gradient and rising prevalence, a possible protective effect of infections in early life, documentation of comorbidities, such as a reduced risk of glioma, and mapping the evidence base through systematic reviews and an online global resource of clinical trials.Ignorance –  Areas where significant uncertainty still exists include the question of whether AD is more than one disease, the tendency for researchers to look at the same old risk factors, lack of specific environmental risk factors that are amenable to manipulation, inconsistencies in the hygiene hypothesis, sparse knowledge about adult AD, lack of evidence that eczema can be prevented, and little scientific work exploring what causes flares in people with established AD.
Comparison of hair follicle histology between horses with pituitary pars intermedia dysfunction and excessive hair growth and normal aged horses
Authors: Innerå Marie , Petersen Annette D. , Desjardins Danielle R. , Steficek Barbara A. , Rosser Edmund J. , Schott Harold C. 
Published: January 2013

Background – Pituitary pars intermedia dysfunction (PPID) in older equids is commonly recognized by a long hair coat that fails to shed.Objective – The aim of this study was to compare hair follicle stages in PPID‐affected horses with excessively long hair coats with the stages of normal aged horses (controls) and to compare hair follicle stages in PPID‐affected horses after 6 months of treatment with pergolide mesylate with those of control horses.Animals – Eight PPID‐affected horses and four normal, age‐matched, control horses.Methods – Skin biopsies were collected from the neck and rump of PPID‐affected and control horses. A diagnosis of PPID was established based on hair coat changes and supportive overnight dexamethasone suppression test results. Skin biopsies were repeated after 6 months of treatment with pergolide. The number of hair follicles in anagen (A) or telogen (T) was counted for each skin biopsy using transverse sections.Results – Pretreatment biopsies had a greater percentage of A follicles (neck 96%, rump 95%) and a lower percentage of T follicles (neck 4%, rump 5%) in PPID‐affected horses than in control horses (A, neck 15%, rump 25%; and T, neck 85%, rump 75%). After treatment with pergolide, all PPID‐affected horses had improved shedding, and the percentages of A follicles (neck 69%, rump 70%) and T follicles (neck 31%, rump 30%) were not different from untreated control horses (A, neck 68%, rump 82%; and T, neck 32%, rump 18%).Conclusions – These findings document that excessive hair growth (hypertrichosis) in PPID‐affected horses is due to persistence of hair follicles in A. Furthermore, treatment with pergolide improved shedding and reduced the percentage of A follicles in PPID‐affected horses.
Kinase dysfunction and kinase inhibitors
Authors: London Cheryl A. 
Published: January 2013

With recent advances in molecular biology, abnormalities in cancer cells that contribute to dysregulation of cell survival and proliferation are being characterized with greater precision. Through this process, key abnormalities in cancer cells involving proteins that regulate signal transduction, migration, mitosis and other critical processes have been identified. Such abnormalities often involve a class of proteins called kinases that act to phosphorylate other proteins in the cell, resulting in activation of these proteins in the absence of appropriate stimulation/regulation. Given their role in tumour biology, substantial effort has been directed at blocking the function of these proteins. Several approaches have been used, including monoclonal antibodies and small molecule inhibitors. While antibodies are primarily directed at cell surface proteins, small molecule inhibitors, also known as kinase inhibitors, target proteins throughout the cell.A variety of kinase inhibitors have been approved for the treatment of human cancers. In some instances, these inhibitors have exhibited significant clinical efficacy, and it is likely that their biological activity will be further enhanced as combination regimens with standard treatment modalities are explored.The use of kinase inhibitors in dogs and cats is relatively recent, although two inhibitors, toceranib (Palladia; Pfizer Animal Health, Madison, NJ, USA) and masitinib (Kinavet; Catalent Pharma Solutions, Somerset, NJ, USA) have been approved by the Federal Drug Administration (USA) for use in dogs. This article reviews the biology of protein kinase dysfunction in human and animal cancers, and the application of specific kinase inhibitors to veterinary cancer patients.
Innate immune defense system of the skin
Authors: Afshar Maryam , Gallo Richard L. 
Published: January 2013

Background –  Antimicrobial peptides (AMPs) have a pivotal role in cutaneous innate immunity. They are present in the skin of many animals, including mammals, and are both constitutively present and inducible by infection and injury.Functions –  Antimicrobial peptides exhibit antimicrobial activity against bacteria, viruses, fungi and parasites, with different potencies depending on their peptide structure. They also act as multifunctional effector molecules that influence diverse cellular processes, including cell migration, proliferation and differentiation, cytokine production, angiogenesis and wound healing. Suppressed AMP production has been associated with increased susceptibility to microbial insults and the pathogenesis of atopic dermatitis.This review highlights recent observations on the expression and role of AMPs, particularly the AMPs cathelicidin and β‐defensin, in healthy and diseased skin.
A systematic review of randomized controlled trials for prevention or treatment of atopic dermatitis in dogs: 2008–2011 update
Authors: Olivry Thierry , Bizikova Petra 
Published: January 2013

Background –  The management of atopic dermatitis (AD) in dogs relies mainly on the use of interventions to reduce pruritus and skin lesions.Objectives –  To provide a critical analysis of recent clinical trials reporting the efficacy and safety of interventions for canine AD.Methods –  Systematic review of randomized controlled trials (RCTs) published, presented or completed between 2008 and 2011, which enrolled dogs with AD. The search was done using electronic databases, reviewing published meeting abstracts and sending queries to professional email lists. Trials reporting the efficacy of interventions aimed at treating, preventing or reducing glucocorticoid usage in atopic dogs were selected.Results –  Twenty‐one RCTs were included. We found further moderate‐quality evidence of efficacy and safety of oral glucocorticoids and ciclosporin for treatment of canine AD. There was additional moderate‐quality evidence of the efficacy of a topical glucocorticoid spray containing hydrocortisone aceponate. Low‐quality evidence was found for the efficacy and safety of injectable recombinant interferons, a budesonide leave‐on conditioner, a ciclosporin topical nano‐emulsion and oral fexofenadine. There is low‐quality evidence of efficacy of oral masitinib, with a need for monitoring for protein‐losing nephropathy. Finally, we uncovered low‐quality evidence of efficacy of a commercial diet as a glucocorticoid‐sparing intervention and of a glucocorticoid spray as a flare‐delaying measure. Very low‐quality evidence was found for the efficacy of other interventions.Conclusions and clinical importance –  Topical or oral glucocorticoids and oral ciclosporin remain the interventions with highest evidence for efficacy and relative safety for treatment of canine AD.