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  • 1
    In: Acta Ophthalmologica, Wiley, Vol. 100, No. S267 ( 2022-01)
    Abstract: A common early lesion of diabetic retinopathy in human patients and a diverse range of animal models is retinal capillary basement membrane thickening occur within 6 months of induction of diabetes. This hallmark pathology of diabetic microvascular disease is used in the evaluation of drug‐mediated modulation of diabetic retinopathy in a range of animal models. Moreover results from published clinical practice have shown that more than 22% of dogs with naturally contracted diabetes mellitus develop ophtalmoscopic signs of retinal hemorrhages or microaneurysms in a relatively short time from onset of the disease. Therefore, we hypothesize that changes in the retina in such dogs results in changes in the protein profile of tear fluid, that can be of the diagnostic or therapeutic value and can contribute to the better understanding the pathophysiology mechanisms underlying that disease. The objective of this study was to identify the protein content of tear film in dogs with naturally contracted diabetes mellitus with and/ without retinopathy, and its comparison with the content of control group. That approach results with mapping out the comprehensive proteome of tear film in diabetic dog and the may contribute to development a noninvasive method to detect potential protein candidates for diabetic retinopathy biomarkers in tear film. Methods Tear film was collected with Schirmer strips from the canine diabetic patients with/ and without retinopathy and sex‐ and age‐matched control group.The study was conducted on 24 dogs (8 dogs in each group).Two‐dimensional electrophoresis was performed followed by MALDI‐TOF mass spectrometry identification of differential proteins. Quantitative analysis of the differentiating electrophoretic spots was done with 2Delta software. Metabolic pathways associated with those proteins were identified with Panther GO software. Results Altogether, 5 significantly differentially expressed proteins were identified. Of those, 4 were downregulated, and 1 was upregulated in the tear film of diabetic patients with retinopathy. Pathway analysis revealed that the proteins were associated with multiple pathways involved in biological regulation, cellular and metabolic process and cellular component organization. Conclusions Tear film protein analysis provides a novel way to screen diabetic retinopathy biomarkers in dogs.
    Type of Medium: Online Resource
    ISSN: 1755-375X , 1755-3768
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2022
    detail.hit.zdb_id: 2466981-7
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  • 2
    In: BMC Veterinary Research, Springer Science and Business Media LLC, Vol. 18, No. 1 ( 2022-12)
    Abstract: Diabetes mellitus (DM) often leads to dangerous thromboembolic complications in humans. DM is also a relatively common endocrinopathy of dogs. There is scarce information regarding procoagulant and anticoagulant plasma indicators in this disease. The aim of the study was to evaluate the levels of the selected plasma haemostatic parameters in dogs suffering from diabetes. The study group consisted of 20 dogs meeting all the inclusion criteria, with fasting glycaemia exceeding 11.1 mmol/l. The control group consisted of 15 healthy dogs presented for routine examination. An evaluation of the prothrombin time (PT); and fibrinogen, D-dimer and antithrombin III (ATIII) levels was performed. Results Except for ATIII activity, the haemostatic parameter differences were not statistically significant. High values of ATIII activity were observed in 90% of diabetic dogs. On average, the values amounted to 166.6% and were 31.4% higher than those in the control group. The ATIII activity in the diabetic group was significantly higher than that in the control group ( p  = 0.0004). Conclusions Here, we report elevated levels of ATIII in diabetic dogs. This finding may suggest the protective role of ATIII against potential thrombotic events. However, the exact role of ATIII in dog diabetes remains unclear.
    Type of Medium: Online Resource
    ISSN: 1746-6148
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2022
    detail.hit.zdb_id: 2191675-5
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  • 3
    Online Resource
    Online Resource
    Public Library of Science (PLoS) ; 2015
    In:  PLOS ONE Vol. 10, No. 12 ( 2015-12-23), p. e0144242-
    In: PLOS ONE, Public Library of Science (PLoS), Vol. 10, No. 12 ( 2015-12-23), p. e0144242-
    Type of Medium: Online Resource
    ISSN: 1932-6203
    Language: English
    Publisher: Public Library of Science (PLoS)
    Publication Date: 2015
    detail.hit.zdb_id: 2267670-3
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  • 4
    Online Resource
    Online Resource
    Wiley ; 2021
    In:  Acta Ophthalmologica Vol. 99, No. S265 ( 2021-01)
    In: Acta Ophthalmologica, Wiley, Vol. 99, No. S265 ( 2021-01)
    Abstract: Diabetes mellitus is becoming an increasing concern in veterinary practice and research. Not treated, it can lead to serious systemic complications and even animal death. The most common form of diabetes in dogs resembles type 1 diabetes in humans. Tear film is easily accessible fluid that can be collected during routine veterinary visit. Based on research conducted in humans, we propose that diabetes can reflect on tear film proteomic composition. Methods The study was conducted on 49 dogs. Enrollment was restricted to clinically stable diabetic dogs. Nineteen healthy age‐ and sex‐matched dogs were included as a control group. Tear film was collected with Schirmer strips from the diabetes and sex‐ and age‐matched control group. Two‐dimensional electrophoresis was performed followed by MALDI‐TOF mass spectrometry identification of differential proteins. Quantitative analysis of the differentiating electrophoretic spots was done with 2Delta software. Metabolic pathways associated with those proteins were identified with Panther GO software. Results Nine out of a total 489 proteins detected on all of the gels were identified as significantly differentially expressed (p ≤ 0.05) in tear film samples collected from diabetic patients. Eight of the nine proteins were assigned to downregulated, namely Phosphatydylinositol‐4 kinase, Pro‐melanin concentrating hormone, Flotillin‐1, Protein mono‐adp ribosyltransferase, ATP‐dependent RNA helicase, GRIP and coiled coil domain containing protein 2, Tetratricopeptide repeat protein 36, Myosin light chain 6B and Prelamin A/C. One of the nine proteins (SRC kinase signalling inhibitor 1) was assigned to upregulated. Conclusions Tear film obtained from dogs with diabetes differs from the healthy controls. Those findings correspond with research conducted in humans, and can add value to our understanding about the course of the disease in dogs. Also, dogs serve well as an animal model of various human diseases.
    Type of Medium: Online Resource
    ISSN: 1755-375X , 1755-3768
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2021
    detail.hit.zdb_id: 2466981-7
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  • 5
    In: Acta Ophthalmologica, Wiley, Vol. 99, No. S265 ( 2021-01)
    Abstract: Age‐related macular degeneration (AMD) is a multifactorial disease, and a leading cause of blindness elderly in the developed countries. In its neovascular form, an immediate treatment is required to maintain the best possible vision. The better understanding of the mechanisms underlying AMD is needed to create a both new treatment protocols and diagnostic tests. Tear film is an easily accessible fluid that can be obtained from each patient during the routine ophthalmic appointment. Changes in tear film proteomic composition have been reported in various ophthalmic and systemic diseases. There is an evidence that the acute form of wet AMD with subretinal fluid may reflect in a tear film composition. Methods Tear film was collected with Schirmer strips from the patients with neovascular AMD and sex‐ and age‐matched control group. Two‐dimensional electrophoresis was performed followed by MALDI‐TOF mass spectrometry identification of differential proteins. Quantitative analysis of the differentiating electrophoretic spots was done with 2 Delta software. Metabolic pathways associated with those proteins were identified with Panther GO software. Results Altogether, 16 differentially expressed proteins were identified. Of those, 14 were downregulated, while 2 showed upregulation in the tear film of wet AMD patients. Pathway analysis revealed multiple pathways involving protein clearance, inflammation, neovascularization and apoptosis. Conclusions Differentially expressed proteins and theirs signaling pathways identified in tear film coincide with impaired protein clearance, persistent inflammation, neovascularization and cell death. Tear film protein analysis provides a novel way to screen AMD‐related biomarkers.
    Type of Medium: Online Resource
    ISSN: 1755-375X , 1755-3768
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2021
    detail.hit.zdb_id: 2466981-7
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  • 6
    In: Animals, MDPI AG, Vol. 12, No. 6 ( 2022-03-16), p. 748-
    Abstract: In this study we aimed to analyze the protein composition of the urine collected from the healthy animals and compare it to the two diabetic groups (DM I normoalbuminuric diabetic dogs; DM II diabetic dogs with microalbuminuria). We tried to identify potential urinary proteins which could be up- or downregulated in diabetic patients even before the appearance of microalbuminuria. Methods: After obtaining urine, we performed two-dimensional electrophoresis, followed by Delta2D software analysis, which allowed for selection and identification with MALDI-TOF spectrometry, statistically significant differentially expressed proteins. Our study revealed 286 common protein spots on 2D gels from the diabetic and control group. From these proteins five were positively identified by MALDI-TOF MS. To further evaluate the five differentiating proteins, the Panther program was used to assign them to appropriate biological process. Conclusion: Significant number of identified proteins play a role in intracellular signaling—vesicle formation, bonding, transport through membranes. This may suggest that first signs of kidney diabetic cellular impairment may be seen in the urine composition before any clinical signs occur.
    Type of Medium: Online Resource
    ISSN: 2076-2615
    Language: English
    Publisher: MDPI AG
    Publication Date: 2022
    detail.hit.zdb_id: 2606558-7
    SSG: 23
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  • 7
    Online Resource
    Online Resource
    MDPI AG ; 2020
    In:  Animals Vol. 10, No. 12 ( 2020-12-17), p. 2416-
    In: Animals, MDPI AG, Vol. 10, No. 12 ( 2020-12-17), p. 2416-
    Abstract: Canine diabetes mellitus is a significant health burden, followed with numerous systemic complications, including diabetic cataracts and retinopathy, leading to blindness. Diabetes should be considered as a disease damaging all the body organs, including gastrointestinal tract, through a complex combination of vascular and metabolic pathologies, leading to impaired gut function. Tear film can be obtained in a non-invasive way, which makes it a feasible biomarker source. In this study we compared proteomic changes ongoing in tear film of diabetic dogs. The study group consisted of 15 diabetic dogs, and 13 dogs served as a control group. After obtaining tear film with Schirmer strips, we performed 2-dimensional electrophoresis, followed by Delta2D software analysis, which allowed to select statistically significant differentially expressed proteins. After their identification with MALDI-TOF (matrix assisted laser desorption and ionisation time of flight) spectrometry we found one up-regulated protein in tear film of diabetic dogs—SRC kinase signaling inhibitor 1 (SRCIN1). Eight proteins were down-regulated: phosphatidylinositol-4 kinase type 2 alpha (PI4KIIα), Pro-melanin concentrating hormone (Pro-MCH), Flotillin-1, Protein mono-ADP ribosyltransferase, GRIP and coiled coil domain containing protein 2, tetratricopeptide repeat protein 36, serpin, and Prelamin A/C. Identified proteins were analyzed by Panther Gene Ontology software, and their possible connections with diabetic etiopathology were discussed. We believe that this is the first study to target tear film proteome in canine diabetes. We believe that combined with traditional examination, the tear film proteomic analysis can be a new source of biomarkers both for clinical practice, and experimental research.
    Type of Medium: Online Resource
    ISSN: 2076-2615
    Language: English
    Publisher: MDPI AG
    Publication Date: 2020
    detail.hit.zdb_id: 2606558-7
    SSG: 23
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  • 8
    Online Resource
    Online Resource
    Springer Science and Business Media LLC ; 2018
    In:  Graefe's Archive for Clinical and Experimental Ophthalmology Vol. 256, No. 6 ( 2018-6), p. 1127-1139
    In: Graefe's Archive for Clinical and Experimental Ophthalmology, Springer Science and Business Media LLC, Vol. 256, No. 6 ( 2018-6), p. 1127-1139
    Type of Medium: Online Resource
    ISSN: 0721-832X , 1435-702X
    RVK:
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2018
    detail.hit.zdb_id: 1459159-5
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  • 9
    In: Acta Ophthalmologica, Wiley, Vol. 100, No. S267 ( 2022-01)
    Abstract: The exact mechanisms underlying the age‐related macular degeneration (AMD) are still not totally understood, but processes like inflammation, chronic oxidative stress (OS), and autophagy impairment, are currently believed to have strong influence of the disease development. There are multiple studies concerning changes of the proteome in different tissues in the course of AMD. As they all need invasive procedures to obtain a sample, we sought for a non‐invasive approach. Tear film can be a potential candidte for proteomic information regarding AMD. Methods Tear film was collected on Schirmer strips, 2D‐electrophoresis was performed, and Delta2D software was used for the identification of the differentially expressed proteins from the gels. The spots were excised, and identified by MALDI‐TOF mass spectrometer. Results 30 patients were examined, 15 for each group. We have found 469 proteins in the analyzed samples, 11 differentiating proteins fulfilled the criteria of being statistically significant. 3 proteins were up‐regulated (Retinal dehydrogenase 1; ATP‐dependent translocase ABCB1; and alpha‐Enolase), and 8 were down‐regulated (Annexin A1; Annexin A4; Glutathione S‐transferase P; Aldo‐ketoreductase family 1 member A1; Allograft inflammatory factor 1; Cytospin‐A or Elongation factor 2; Short stature homeobox protein 2 and uncharacterized protein C11orf98). Conclusions Tear film can be used as a non‐invasively obtained source of proteomic information about cellular processes ongoing in the course of AMD. The differentially expressed proteins are involved in the choroidal neovascularization, inflammation, oxidative stress and autophagy impairment.
    Type of Medium: Online Resource
    ISSN: 1755-375X , 1755-3768
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2022
    detail.hit.zdb_id: 2466981-7
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  • 10
    In: Journal of Clinical Medicine, MDPI AG, Vol. 10, No. 14 ( 2021-07-10), p. 3060-
    Abstract: Macular edema and its further complications due to the leakage from the choroidal neovascularization in course of the age-related macular degeneration (AMD) is a leading cause of blindness among elderly individuals in developed countries. Changes in tear film proteomic composition have been reported to occur in various ophthalmic and systemic diseases. There is an evidence that the acute form of neovascular AMD may be reflected in the tear film composition. Tear film was collected with Schirmer strips from patients with neovascular AMD and sex- and age-matched control patients. Two-dimensional electrophoresis was performed followed by MALDI-TOF mass spectrometry for identification of differentially expressed proteins. Quantitative analysis of the differential electrophoretic spots was performed with Delta2D software. Altogether, 11 significantly differentially expressed proteins were identified; of those, 8 were downregulated, and 3 were upregulated in the tear film of neovascular AMD patients. The differentially expressed proteins identified in tear film were involved in signaling pathways associated with impaired protein clearance, persistent inflammation, and neovascularization. Tear film protein analysis is a novel way to screen AMD-related biomarkers.
    Type of Medium: Online Resource
    ISSN: 2077-0383
    Language: English
    Publisher: MDPI AG
    Publication Date: 2021
    detail.hit.zdb_id: 2662592-1
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