Primary structure of potato Kunitz-type serine proteinase inhibitor

The serine proteinase inhibitor (PSPI-51) isolated from potato tubers (Solanum tuberosum L,) comprises two protein species with pi 5.2 and 6.3, denoted as PSPI-21-5.2 and PSPI-21-6.3, respectively. They were separated by anion exchange chromatography on a Mono Q FPLC column. Both species tightly inhibit human leukocyte elastase, whereas their interaction with trypsin and chymotrypsin is substantially weaker. The sequences of both PSPI-21-5.2 and PSPI-21-6.3 were determined by analysis of overlapping peptides obtained from the oxidized or reduced and S-pyridylethylated proteins after digestion with trypsin or pepsin, Both species of PSPI-21 are composed of two chains, named chains A and B, which are linked by a disulfide bridge between Cys(146) and Cys(157). The other disulfide bridge is located within the A chains between Cys(48) and Cys(97). The amino acid sequences of the large A chains of the two forms, consisting of 150 amino acids residues each, differ in a single residue at position 52. The small chains B, containing 37 and 36 residues in PSPI-21-6.3 and PSPI-21-5.2, respectively, have nine different residues. The entire amino acid sequences of the two inhibitors show a high degree of homology to the other Kunitz-type proteinase inhibitors from plants

Comparative Analyses of Exoproteinases Produced by Three Phytopathogenic Microorganisms

Proteinases secreted by the oomycete Phytophthora infestans (Mont.) de Bary, Rhizoctonia solani, and Fusarium culmorum belonging to different families of fungi have been studied to determine if the exoenzyme secretion depends on the environmental conditions and the phylogenetic position of the pathogen. The substrate specificity of the extracellular proteinases of F. culmorum, R. solani, and P. infestans and their sensitivity to the action of synthetic and protein inhibitors suggest that they contain trypsin-like and subtilisin-like enzymes regardless of culture medium composition. The relation of trypsin-like and subtilisin-like enzymes is dependent on the culture medium composition, especially on the form of nitrogen nutrition, particularly in the case of the exoenzymes secreted by R. solani. Phylogenetic analyses have shown that the exoproteinase set of ascomycetes and oomycetes has more similarities than basidiomycetes although they are more distant relatives. Our data suggests that the multiple proteinases secreted by pathogenic fungi could play different roles in pathogenesis, increasing the adaptability and host range, or could have different functions in survival in various ecological habitats outside the host

Physical and chemical processes and the morphofunctional characteristics of human erythrocytes in hyperglycaemia

Background: This study examines the effect of graduated hyperglycaemia on the state and oxygen-binding ability of hemoglobin, the correlation of phospholipid fractions and their metabolites in the membrane, the activity of proteolytic enzymes and the morphofunctional state of erythrocytes. Methods: Conformational changes in the molecule of hemoglobin were determined by Raman spectroscopy. The structure of the erythrocytes was analyzed using laser interference microscopy (LIM). To determine the activity of NADN-methemoglobinreductase, we used the P.G. Board method. The degree of glycosylation of the erythrocyte membranes was determined using a method previously described by Felkoren et al. Lipid extraction was performed using the Bligh and Dyer method. Detection of the phospholipids was performed using V. E. Vaskovsky method. Results: Conditions of hyperglycaemia are characterized by a low affinity of hemoglobin to oxygen, which is manifested as a parallel decrease in the content of hemoglobin oxyform and the growth of deoxyform, methemoglobin and membrane-bound hemoglobin. The degree of glycosylation of membrane proteins and hemoglobin is high. For example, in the case of hyperglycaemia, erythrocytic membranes reduce the content of all phospholipid fractions with a simultaneous increase in lysoforms, free fatty acids and the diacylglycerol (DAG). Step wise hyperglycaemia in incubation medium and human erythrocytes results in an increased content of peptide components and general trypsin-like activity in the cytosol, with a simultaneous decreased activity of µ-calpain and caspase 3. Conclusions: Metabolic disorders and damage of cell membranes during hyperglycaemia cause an increase in the population of echinocytes and spherocytes. The resulting disorders are accompanied with a high probability of intravascular haemolysis.</p

Medicinal Ethnobotany and Potential of Medicine Plants of Anak Rawa Ethnic at The Penyengat Village Sungai Apit Siak Riau

Anak Rawa ethnic is indigenous ethnic in the Penyengat Village which is located on the edge of swamp river and forest, so their dependence and adaptation to forest resources is very high. They uses the forest to make medicine plants. The objective of this research was to identify the ethnobotanical knowledge of Anak Rawa ethnic about medicine plant and identify potential species of medicine plants. The research was conducted  in March to April 2018 The potential of plant data was obtained through quantitative methods in the form of vegetation and exploration analysis. Meanwhile, the community knowledge data collected through semi-structured interviews. The research identified 194 medicinal plant species at Penyengat village in several types of habitat (forests, yards, gardens, and roadsides), with 140 medicinal plant species used by Anak Rawa ethnic. There were 54 species of medicinal plants potential in Penyengat Village. Medicine plants used by the community are grouped into 28 medicinal benefits. The most widely used medicinal plants in society are in the Rubiaceae family of 11 species. Keywords: Anak Rawa ethnic, ethnobotany, medicinal plant, potentia

Efektivitas Ekstrak Daun Mangkokan terhadap Penyembuhan Luka Bakar pada Tikus

Luka&nbsp; bakar adalah bentuk kerusakan dan/atau kehilangan jaringan akibat kontak dengan sumber panas dan suhu tinggi (seperti api, air panas, dll) atau suhu yang sangat rendah. Kini sedang dikembangkan terapi luka bakar melalui pemberian topikal ekstrak herbal. Daun Mangkokan (Nothopanax scutellarium) adalah salah satu herbal yang dapat digunakan untuk terapi luka bakar. &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Penelitian ini bertujuan untuk mengetahui efektivitas ekstrak daun Mangkokan dalam bentuk sediaan salep terhadap penyembuhan luka bakar pada hewan uji yaitu tikus putih. Penelitian ini dimulai dengan melakukan proses ekstraksi daun Mangkokan (Nothopanax scutellarium), kemudian dilakukan skrining fitokimia untuk mengidentifikasi alkaloid, flavonoid, saponin, dan tanin. Lalu dibuat salep ekstrak daun Mangkokan (Nothopanax scutellarium) dengan konsentrasi 50% dan 75%. Hewan uji yaitu tikus putih, sebelumnya sudah dipersiapkan selama 1 minggu sehingga hewan uji mampu beradaptasi. Dilakukan pembuatan luka bakar pada bagian punggung kanan tikus. Selanjutnya dilakukan penyembuhan luka bakar, disiapkan 4 kelompok hewan uji yaitu: K1: tidak diberi perlakuan (kontrol negatif); K2: Diberikan sediaan bioplacenton (kontrol positif); K3: diberikan sediaan salep ekstrak daun Mangkokan konsentrasi 50%; K4: diberikan sediaan salep ekstrak daun Mangkokan konsentrasi 75%. Pengolesan dilakukan secara merata 2 kali sehari, pagi dan sore, kemudian ditutup dengan kassa steril. Luka diamati selama 21 hari, setiap 3 hari sekali dengan mengamati secara makroskopik. Berdasarkan hasil pengujian senyawa kimia terbukti bahwa ekstrak daun Mangkokan mengandung senyawa alkaloid, saponin, flavonoid dan tannin. Berdasarkan hasil penelitian pada hari ke-21, pada kelompok KI keropeng sudah terkelupas semua namun daerah yang terkelupas masih terlihat basah, bentuk ukuran mengecil, daerah bekas luka masih terlihat kotor. Pada kelompok K2 dan K3, keropeng sudah terkelupas dan kering, ukuran mengecil, dan pada K4 keropeng sudah tidak terlihat, warna hampir merata dan ukuran sudah jauh lebih mengecil, bekas luka lebih bersih. Hasil penelitian ini menunjukan bahwa salep ekstrak daun Mangkokan mempercepat penyembuhan luka bakar pada tikus putih