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Urine proteome from healthy and CKD cats
No full textIn veterinary medicine there is an increasing interest in the application of proteomic techniques to investigate protein patterns in healthy and diseased animals; however, data on urine proteome are still limited (1,2,3). The aims of our study were to identify a urine protein profile in healthy cats and to compare it with those obtained in patients affected by chronic kidney disease (CKD). Urine samples were collected by cystocentesis from 23 healthy and 18 CKD cats. For all samples urinalysis and urine protein to creatinine ratio (UPC) were performed. Urine proteins were further separated by SDS-PAGE and the bands were reduced, alkylated and then digested by trypsin before ESI-Q-TOF mass spectrometry analysis. Protein identification was performed using MASCOT science search engine. Healthy cats had significantly (p<0.01) lower values of UPC than CKD cats. SDS-PAGE allowed to visualize an “healthy profile” with many different bands (median 32; range 26-47), including Albumin (70 kDa), Cauxin (carboxylesterase 5A, 61 kDa), Uromodulin (73 kDa), Transferrin (80 kDa), Angiotensin-converting enzyme 2 (93 kDa), Inter-alpha-trypsin inhibitor heavy chain H4 (103 kDa) and α2Macroglobulin (170 kDa); at lower molecular weights Albumin- (55kDa) and Cauxin- (40kDa) fragments, Haptoglobin (45kDa) and immunoglobulin light chains (24kDa) were present. Retinol binding protein 4 (23 kDa) and Cystatin-M(16 kDa) were identified only in urine of CKD patients.
Proteomic techniques were successfully used to investigate proteinuria, indicating that two proteins are differentially expressed in urine of healthy and CKD cats. These proteins could be considered as promising biomarkers of chronic renal damage in feline patients.
References
1. Lemberger SI, Deeg CA, Hauck SM, Amann B, Hirmer S, Hartmann K, Dorsh R. Comparison of urine protein profiles in cats without urinary tract disease and cats with idiopathic cystitis, bacterial urinary tract infection, or urolithiasis. Am J Vet Res. 2011;72(10):1407-15.
2. Schaefer H, Kohn B, Schweigert FJ, Raila J. Quantitative and qualitative urine protein excretion in dogs with severe inflammatory response syndrome. J Vet Intern Med. 2011;25(6):1292-7.
3. Nabity MB, Lees GE, Dangott LJ, Cianciolo R, Suchodolski JS, Steiner JM. Proteomic analysis of urine from male dogs during early stages of tubulointerstitial injury in a canine model of progressive glomerular disease. Vet Clin Pathol. 2011;40(2):222.
Figures
Figure 1. SDS-PAGE of urine sampples from healthy and CKD cats stained with silver staining. a) Electrophoretic profiles: 1 MW marker; 2-6 CKD urine samples; 7-8, pools of healthy urine samples female and male respectively; b) Pherograms obtained by ImageJ software
Proteomic analysis of urine from healthy and CKD cats
No full textIn veterinary medicine there is an increasing interest in the application of proteomic techniques to investigate protein patterns in healthy and diseased animals; however, data on urine proteome are still limited. The aims of our study were to identify a urine protein profile in healthy cats and to compare it with those obtained in patients affected by chronic kidney disease (CKD). Urine samples were collected by cystocentesis from 23 healthy and 18 CKD cats. For all samples urinalysis and urine protein to creatinine ratio (UPC) were performed. Urine proteins were further separated by SDS-PAGE and the bands were reduced, alkylated and then digested by trypsin before ESI-Q-TOF mass spectrometry analysis. Protein identification was performed using MASCOT science search engine. Healthy cats had significantly (p<0.01) lower values of UPC than CKD cats. SDS-PAGE allowed to visualize an “healthy profile” with many different bands (median 32; range 26-47), including Albumin (70 kDa), Cauxin (carboxylesterase 5A, 61 kDa), Uromodulin (73 kDa), Transferrin (80 kDa), Angiotensin-converting enzyme 2 (93 kDa), Inter-alpha-trypsin inhibitor heavy chain H4 (103 kDa) and α2Macroglobulin (170 kDa); at lower molecular weights, Albumin- (55kDa) and Cauxin- (40kDa) fragments, Haptoglobin (45kDa) and immunoglobulin light chains (24kDa) were present. Retinol binding protein 4 (23 kDa) and Cystatin-M(16 kDa) were identified only in urine of CKD patients.
Proteomic techniques were successfully used to investigate proteinuria, revealing different protein patterns between healthy and CKD cats. Some of these proteins could be considered as promising biomarkers of chronic renal damage in feline patients
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
A 2DE map of the urine proteome in the cat: effect of Chronic Kidney Disease
No full textIntroduction
Urine is considered an ideal source of clinical biomarkers as it can be obtained noninvasively, repeatedly and in adequate amounts. In veterinary medicine, the application of proteomics techniques is still very limited. The aim of our work was to produce a preliminary map of the urine proteome of the healthy cats (Felis catus) and to compare it with the proteome of cats affected by chronic kidney disease (CKD). For that we have used an approach based on two-dimensional electrophoresis and protein identification using mass spectrometry (MS).
Materials and Methods
Urine samples were collected by cystocentesis from 4 healthy and 4 cats affected by CKD and analyzed by 2DE. The first dimension was performed by isoelectric focusing on 17 cm long IPG strips (pH 3-10); the second dimension was performed on 10% SDS-PAGE and stained with colloidal Coomassie. Spots were excised from the gel, reduced, alkylated and digested with trypsin and identified using ESI-Q-TOF MS (Campos et al., 2013).
Results and Discussions
2DE allowed the separation of 66 spots in the urine proteome of healthy and CKD cats. Eighteen spots were overrepresented in CKD and nine spots were underrepresented. The 27 differentially expressed spots and the nine most abundant common spots were excised from the gels for MS identification (Figure 1; Table 1).
Preliminary 2D map. 21 spots yielded significant results by MS, producing a preliminary feline urine map, including 13 proteins that may be functionally classified as transport (38%), immune and cellular response (38%), and cellular communication and growth (15%). The most abundant protein was cauxin, a serine esterase produced by healthy tubular cells, specifically excreted in urine of cats and probably involved in the synthesis of felinine pheromone (Miyazaki et al., 2007). The transport proteins, albumin, transferrin, haemopexin and haptoglobin all derive from plasma and have been identified as common components of urine also from healthy humans (Candiano et al., 2010). Among the proteins involved in immune and cellular defence response, we identified IgK light chain, protein AMBP and uromodulin. Differently from dogs (Brandt et al., 2014) and humans (Lhotta, 2010), uromodulin is not the most abundant urine-specific protein in cats. The remaining proteins, perlecan and fetuin-A, are involved in cell communication and growth. In particular, perlecan, a negatively charged proteoglycan of the glomerular filtration barrier, has also been identified in dog urine (Nabity et al., 2011).
Effect of CKD. Regarding the effect of CKD on the urine proteome, seven differentially represented proteins have been identified These proteins can be indicative of tubular dysfunction when not reabsorbed (e.g. RBP) or not secreted (e.g. uromodulin and cauxin) and could be studied as putative biomarkers of nephropathy. Among the overrepresented proteins, retinol binding protein (RBP) is a 22 kDa protein freely filtered by the glomerulus and reabsorbed by the tubules. The appearance of RBP in urine is a marker of impaired tubular function in humans (Pallet et al., 2014) and it has also been reported in dogs (Nabity et al., 2011) and cats (van Hoek et al., 2008). Interesting underrepresented proteins were uromodulin and cauxin. Uromodulin is a glycoprotein produced by healthy tubular cells and its disappearance has been already proved in humans affected by CKD (Lhotta, 2010) and could be applied also in cats. Regarding cauxin, according to Miyazaki et al., (2007) this protein could be a promising biomarker for the determination of tubular damage in CKD cats.
Conclusions
2DE was essential in fractionation of the complex urine proteome in cats, producing a preliminary map that included 13 proteins. In particular, uromodulin, cauxin and perlecan, specifically secreted in urine, could help in the evaluation of renal function. Seven proteins were differentially represented in CKD cats suggesting their use as a putative biomarker of nephropathy in cats and possibly in other veterinary species
Dispelling the Myths Behind First-author Citation Counts
Full text linkWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
koamabayili/VECTRON-author-checklist: VECTRON author checklist
No full textWe have done our best to complete the author checklist relating to the use of animals in the hut study. Note that the objective for the hut study was to evaluate the IRS treatment applications for residual efficacy against Anopheles mosquitoes, including the local An. coluzzii mosquito population. Cows were only used to attract mosquitoes into the huts and no tests were carried out directly on the cows. The author checklist is intended for use with studies where experiments are carried out on animals, which is why we have had such difficulty in completing this for the hut study, as many of the questions do not relate to how the cows were used
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