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Background Tear proteomic analysis has become an important tool in medical and veterinary research. The tear collection method could influence the tear protein profile. This study aims to evaluate the protein profiles of dog tears collected using microcapillary tubes (MT), Schirmer tear strips (ST), and ophthalmic sponges (OS). Methods The tear samples were collected using MT, ST, and OS. Tear protein profiles were analyzed using two-dimensional electrophoresis (2-DE) and the different protein spots’ expression was compared. Fourteen protein spots were identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results Tear protein concentrations ranged from 2.80 to 4.03 μg/μL, with no statistically significant differences among collection methods. Protein expression in each collection method differed in terms of both the number and intensity of the spots. There were 249, 327, and 330 protein spots found from tears collected with MT, ST, and OS, respectively. The proteins albumin, haptoglobin, and lactoferrin identified from OS were found to have higher spot intensities than other methods of collection. The use of MT demonstrated the downregulation of nine proteins. Conclusions The recent study supported that tear protein analysis is affected by different tear collection methods. Although ST is commonly used for tear collection, it provides insufficient information to study particular tear proteins.

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Peptidomic analysis coupled with mass spectrometry identification was employed to investigate the constitutively expressed peptides in latex serum Phytophthora-resistant rubber tree (Hevea brasiliensis), BPM24. The utilization of proteins and peptides as a strategy to study new methods can lead to a therapeutic application. However, a high-efficiency method to extract peptides from latex serum is not well established. After optimization of latex serum isolation from natural rubber latex, using of lysis solution for peptide extraction exhibited superior serum peptide recovery than using other chemical reagents. Ultrafiltration followed by solid-phase extraction was selected to achieve high extraction yield of low molecular weight proteins and peptides. Mass spectrometric and bioinformatics analysis characterized and predicted the identified protein and peptide contents. All latex serum peptide fractions from C18 reversed-phase chromatography possessed effectiveness of antifungal activity against three tested plant fungal including A. brassicicola, C. lunata and M. grisea with different MIC in the range 250-2,000 μg/ml from different peptide fraction. Moreover, Etotal (E1-E5 pooled together) from lysis solution extraction was found to able to inhibit three tested plant fungal at MIC 500 μg/ml. Moreover, B. cereus and E. faecium were the sensitive gram-positive bacteria with MIC in the range 250-4,000 μg/ml. However, all gram-negative bacteria were not affected by 4,000 μg/ml. Among these tested pathogens, plant fungal was quite sensitive to our peptide product. Etotal from lysis solution extraction will be the best candidate for using further in antifungal testing of other plant fungal both in vitro and in vivo. The further investigation and application will lead to possible use of latex serum as a great biotechnological resource to develop peptide –based product for the treatment of plant infection.

Background:: Hevea brasiliensis is severely affected by the fungal disease caused by Phytophthora spp. Significant loss of rubber yield is widespread and extensive use of chemical fungicides has resulted in health and environmental problems. background: Hevea brasiliensis is severely affected by the fungal disease caused by Phytophthora spp. Significant loss of rubber yield is widespread and extensive use of chemical fungicides has resulted in health and environmental problems. Objective:: This work aims to extract and identify the latex serum peptides from a disease tolerant clone of Hevea brasiliensis, and study the inhibitory efficacy against pathogenic bacteria and fungi. objective: This work aims to extract and identify the latex serum peptides from a disease tolerant clone of Hevea brasiliensis, and study the inhibitory efficacy against pathogenic bacteria and fungi. Methods:: Serum peptides were extracted from H. brasiliensis BPM 24 using mixed lysis solution. Low molecular weight peptides were screened and fractionated by solid-phase extraction and then identified by tandem mass spectrometry. Total and fractionated serum peptides were assayed for bacterial and fungal inhibition using broth microdilution and poisoned food methods. An inhibitory control study in the greenhouse was also performed using susceptible clones for pre- and post-infection with Phytophthora spp. method: Serum peptides were extracted from H. brasiliensis BPM 24 using mixed lysis solution. Low molecular weight peptides were screened and fractionated by solid-phase extraction and then identified by tandem mass spectrometry. Total and fractionated serum peptides were assayed for bacterial and fungal inhibition using broth microdilution and poisoned food methods. An inhibitory control study in greenhouse was also performed using susceptible clones for pre- and post-infection with Phytophthora spp. Results:: Forty-three serum peptide sequences were successfully identified. Thirty-four peptides matched with the proteins associated with plant defense response signaling, host resistance, and adverse environmental factors. The inhibitory study of total serum peptides demonstrated anti-bacterial and anti-fungal properties. The greenhouse study exhibited disease inhibitory efficacy of 60% for the treatment of Phytophthora spp. in post-infected plants and 80% for pre-treated samples. Conclusion:: Latex serum peptides from disease tolerant H. brasiliensis revealed several proteins and peptides associated with plant defense and disease resistance. The peptides play a vital role for defense against bacteria and fungi pathogens, including Phytophthora spp. Enhanced disease protection can be obtained when the extracted peptides were applied to the susceptible plants before exposure to the fungi. These findings provided an insight and may pave the way for the development of biocontrol peptides from natural resources. other: -