Sample records for chymopapain

  1. Effects of pH on the association between the inhibitor cystatin and the proteinase chymopapain. (United States)

    Reyes-Espinosa, Francisco; Arroyo-Reyna, Alfonso; Garcia-Gutierrez, Ponciano; Serratos, Iris N; Zubillaga, Rafael A


    Cysteine proteinases are involved in many aspects of physiological regulation. In humans, some cathepsins have shown another function in addition to their role as lysosomal proteases in intracellular protein degradation; they have been implicated in the pathogenesis of several heart and blood vessel diseases and in cancer development. In this work, we present a fluorometric and computational study of the binding of one representative plant cysteine proteinase, chymopapain, to one of the most studied inhibitors of these proteinases: chicken cystatin. The binding equilibrium constant, Kb, was determined in the pH range between 3.5 and 10.0, revealing a maximum in the affinity at pH 9.0. We constructed an atomic model for the chymopapain-cystatin dimer by docking the individual 3D protein structures; subsequently, the model was refined using a 100 ns NPT molecular dynamics simulation in explicit water. Upon scrutiny of this model, we identified 14 ionizing residues at the interface of the complex using a cutoff distance of 5.0 Å. Using the pKa values predicted with PROPKA and a modified proton-linkage model, we performed a regression analysis on our data to obtain the composite pKavalues for three isoacidic residues. We also calculated the electrostatic component of the binding energy (ΔGb,elec) at different pH values using an implicit solvent model and APBS software. The pH profile of this calculated energy compares well with the experimentally obtained binding energy, ΔGb. We propose that the residues that form an interchain ionic pair, Lys139A from chymopapain and Glu19B from cystatin, as well as Tyr61A and Tyr67A from chymopapain are the main residues responsible for the observed pH dependence in the chymopapain- cystatin affinity.

  2. Surface Plasmon Resonance Imaging biosensor for cystatin determination based on the application of bromelain, ficin and chymopapain

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    Anna Sankiewicz


    Full Text Available A Surface Plasmon Resonance Imaging (SPRI sensor based on bromelain or chymopapain or ficin has been developed for specific cystatin determination. Cystatin was captured from a solution by immobilized bromelain or chymopapain or ficin due to the formation of an enzyme-inhibitor complex on the biosensor surface. The influence of bromelain, chymopapain or ficin concentration, as well as the pH of the interaction on the SPRI signal, was investigated and optimized. Sensor dynamic response range is between 0–0.6 μg/ml and the detection limit is equal to 0.1 μg/ml. In order to demonstrate the sensor potential, cystatin was determined in blood plasma, urine and saliva, showing good agreement with the data reported in the literature.

  3. Drug: D03483 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available : M09AB01 cysteine protease [EC:] Anatomical Therapeutic Chemical (ATC) classification [BR:br08303] M MUSCULO-SKELETAL... SYSTEM M09 OTHER DRUGS FOR DISORDERS OF THE MUSCULO-SKELETAL SYSTEM M09A OTHER DRUGS FOR ...DISORDERS OF THE MUSCULO-SKELETAL SYSTEM M09AB Enzymes M09AB01 Chymopapain D03483 Chymopapain (USAN/INN) CAS: 9001-09-6 PubChem: 17397619 ...

  4. AcEST: DK953330 [AcEST

    Lifescience Database Archive (English)

    Full Text Available Arabidopsis ... 167 5e-41 sp|P14080|PAPA2_CARPA Chymopapain OS=Carica papaya PE=1 SV=2 158 2e-38 sp|Q94B08|GCP1_ARATH Germination...VSVSEEELVTCSSES-GCDGG 606 TGNL+ +SE+ELV C S GC GG Sbjct: 175 VTGNLLELSEQELVDCDKHSYGCKGG 200 >sp|Q94B08|GCP1_ARATH Germination

  5. Percutaneous treatment of lumbar disc herniation

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    Leonardi, M.; Resta, F.; Bettinelli, A. [Ospedale Maggiore di Milano (Italy); Lavaroni, A.; Fabris, G. [Ospedale Civile di Udine (Italy); Abelli, F. [Fondazione Clinica del Lavorno, Pavia (Italy)


    918 patients were subjected to percutaneous treatment of lumbar herniated disc. 733 of these were treated through enzymatic nucleolysis with chymopapain, for a total 883 levels. Chemonucleolysis was carried out on 733 patients (79.8%). Automated discectomy has been carried out in 185 cases (20.1%). Chemonucleolysis has led to the resolution of the symptoms in a high percentage of cases (82.2%). Nucleotomy has proved to be an effective method in 74.6% of cases. (author). 7 refs, 6 figs, 1 tab.

  6. Effect of spondylosis on the outcome of chemonucleolysis

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    Tehranzadeh, J.; Kaufman, H.H.; Davis, C.M.; Radin, E.L.


    In an attempt to understand the relation between the outcome of chemonucleolysis treatment of lumbar discs and the presence of spondylosis (degenerative process) of the lumbar spine, 40 cases of disc herniation with subsequent intradiscal chymopapain injection (chemonucleolysis) were reviewed. Thirty-eight patients had plain radiographs, 25 had myelography, 28 had computed tomography (CT), and 15 had both myelography and CT. A review of these examinations revealed that 24 patients had minimal or mild spondylosis (group 1) and 13 patients had moderate spondylosis (group 2). Only one patient had a severe spondylosis (group 3). Chemonucleolysis failed in 8/24 (33%) patients from the first group and 2/13 (15%) cases from the second group. Therefore, the presence or absence of even moderate spondylosis did not affect the treatment success rate. A review of the 11 failed cases revealed spinal canal stenosis in three cases and a large disc herniation in two (one case with a myelographic block). Another case had a sequestered disc fragment. In two further cases, the disc herniation was far lateral with complete occlusion of the lateral foramen; another case exhibited severe degenerative changes of facet joints. In two other cases, treatment failure could not be explained by any radiographic finding. Of three patients with large disc herniation, only one responded successfully to intradiscal injection of chymopapain. Chemonucleolysis, however, failed in the only patient in the third group who had severe spondylosis of the facet joint.

  7. Investigation on Enzyme Clotting Technology of Soft Cheese%软质干酪的酶凝乳工艺研究

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    吕斯思; 陈有亮; 金鸟君


    In order to study the best cheese production process of substitute clotting enzymes, L9(34)and L16(43) designs of experiments on calf rennet ,chymopapain and Mucor miehei chymosin combinations were applied to determine the best technological parameters on cheese production. The results show that the optimum parameters of single enzymes are:calf rennet 0.0125%,temperature 32 ℃,clotting time 4Smin;chymopapain 0.01%,temperature 33 ℃,clotting time 35 min; Mucor miehei chymosin 0.0035%,temperature 32 ℃,clotting time 40 min. The best technological parameters of enzyme combinations are: calf rennet 0.004%, chymopapain 0.006%,Mucor miehei chymosin 0.0023%,temperature 32 ℃,clot-ting time 30 min,which can effectively optimize cheese production process.%为了探求软质干酪的最佳酶凝乳工艺,采用L9(34)和L16(45)正交试验,研究了皱胃酶、木瓜酶、米黑毛霉酶三种单酶的最佳工艺以及复配酶最佳工艺,确定了皱胃酶添加量0.0125%,凝乳温度32℃,时间45min;木瓜酶添加量0.01%,凝乳温度33℃,时间35min;米黑毛霉酶添加量0.0035%,凝乳温度32C,时间40min;复配酶最佳工艺参数为皱胃酶用量0.004%,木瓜酶用量0.006%,米黑毛霉酶用量0.0023%,温度32℃,凝乳时间30min.结果表明,单酶复配具有协同作用,能够有效优化干酪酶凝乳工艺.

  8. Insight to structural subsite recognition in plant thiol protease-inhibitor complexes : Understanding the basis of differential inhibition and the role of water

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    Mukhopadhayay Bishnu P


    Full Text Available Abstract Background This work represents an extensive MD simulation / water-dynamics studies on a series of complexes of inhibitors (leupeptin, E-64, E-64-C, ZPACK and plant cysteine proteases (actinidin, caricain, chymopapain, calotropin DI of papain family to understand the various interactions, water binding mode, factors influencing it and the structural basis of differential inhibition. Results The tertiary structure of the enzyme-inhibitor complexes were built by visual interactive modeling and energy minimization followed by dynamic simulation of 120 ps in water environment. DASA study with and without the inhibitor revealed the potential subsite residues involved in inhibition. Though the interaction involving main chain atoms are similar, critical inspection of the complexes reveal significant differences in the side chain interactions in S2-P2 and S3-P3 pairs due to sequence differences in the equivalent positions of respective subsites leading to differential inhibition. Conclusion The key finding of the study is a conserved site of a water molecule near oxyanion hole of the enzyme active site, which is found in all the modeled complexes and in most crystal structures of papain family either native or complexed. Conserved water molecules at the ligand binding sites of these homologous proteins suggest the structural importance of the water, which changes the conventional definition of chemical geometry of inhibitor binding domain, its shape and complimentarity. The water mediated recognition of inhibitor to enzyme subsites (Pn...H2O....Sn of leupeptin acetyl oxygen to caricain, chymopapain and calotropinDI is an additional information and offer valuable insight to potent inhibitor design.

  9. Chemonucleolysis in lumbar disc herniation: a meta-analysis Quimonucleólise em hernia de disco lombar: metanálise

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    José Mauro Cardoso Couto


    Full Text Available PURPOSE: To carry out a systematic review and meta-analysis of the efficacy of chemonucleolysis in the treatment of lumbar disc herniation. METHODS: Clinical trials were selected from 3 electronic databases (The Cochrane Controlled Trials Register, MEDLINE, and EMBASE. Data were analyzed with the software STATA, using the meta command. RESULTS: Twenty-two clinical trials were eligible. For chemonucleolysis versus placebo, the summary risk ratio estimate for pain relief as outcome was 1.51 (95% CI: 1.27-1.80. The summary estimate was 1.07 (95% CI: 0.95-1.20 for the comparison between chymopapain and collagenase. Regarding chemonucleolysis with chymopapain versus surgery, the fixed-effect summary estimate of effect for pain relief was 0.93 (95% CI: 0.88-0.98 with surgery as the reference group. In this case, heterogeneity was statistically significant. CONCLUSIONS: Chemonucleolysis with chymopapain was superior to placebo and was as effective as collagenase in the treatment of lumbar disc prolapse. Results for studies comparing chemonucleolysis with surgery were heterogeneous, making it difficult to interpret the summary measure of effect.OBJETIVO: Avaliar a eficácia da quimonucleólise no tratamento da hérnia de disco lombar por meio de uma metanálise de ensaios clínicos. MÉTODOS: Os ensaios clínicos foram selecionados de três bases de dados eletrônicas( Cochrane, MEDLINE, e EMBASE. Os dados foram analisados por intermédio do aplicativo STATA, com o comando meta. RESULTADOS: trabalhamos com 22 ensaios clínicos. Para a comparação entre quimonucleólise e placebo, a estimativa da razão de riscos, tendo melhora da dor como desfecho, foi de 1,51 (I 95% C: 1,27-1,80. Aquela medida foi de 1,07 (I 95% C: 0,95-1,20 para a comparação entre quimopapaína e colagenase. Em um modelo de efeitos fixos, a razão de risco, para melhora da dor, foi 0,93 (I 95% C: 0,88-0,98, tendo a discectomia como grupo de referência. Nesse caso, um teste de

  10. A chestnut seed cystatin differentially effective against cysteine proteinases from closely related pests. (United States)

    Pernas, M; Sánchez-Monge, R; Gómez, L; Salcedo, G


    Cystatin CsC, a cysteine proteinase inhibitor from chestnut (Castanea sativa) seeds, has been purified and characterized. Its full-length cDNA clone was isolated from an immature chestnut cotyledon library. The inhibitor was expressed in Escherichia coli and purified from bacterial extracts. Identity of both seed and recombinant cystatin was confirmed by matrix-assisted laser desorption/ionization mass spectrometry analysis, two-dimensional electrophoresis and N-terminal sequencing. CsC has a molecular mass of 11,275 Da and pI of 6.9. Its amino acid sequence includes all three motifs that are thought to be essential for inhibitory activity, and shows significant identity to other phytocystatins, especially that of cowpea (70%). Recombinant CsC inhibited papain (Ki 29 nM), ficin (Ki 65 nM), chymopapain (Ki 366 nM), and cathepsin B (Ki 473 nM). By contrast with most cystatins, it was also effective towards trypsin (Ki 3489 nM). CsC is active against digestive proteinases from the insect Tribolium castaneum and the mite Dermatophagoides farinae, two important agricultural pests. Its effects on the cysteine proteinase activity of two closely related mite species revealed the high specificity of the chestnut cystatin.

  11. Label-free quantitative proteomics reveals differentially regulated proteins in the latex of sticky diseased Carica papaya L. plants. (United States)

    Rodrigues, Silas P; Ventura, José A; Aguilar, Clemente; Nakayasu, Ernesto S; Choi, HyungWon; Sobreira, Tiago J P; Nohara, Lilian L; Wermelinger, Luciana S; Almeida, Igor C; Zingali, Russolina B; Fernandes, Patricia M B


    Papaya meleira virus (PMeV) is so far the only described laticifer-infecting virus, the causal agent of papaya (Carica papaya L.) sticky disease. The effects of PMeV on the laticifers' regulatory network were addressed here through the proteomic analysis of papaya latex. Using both 1-DE- and 1D-LC-ESI-MS/MS, 160 unique papaya latex proteins were identified, representing 122 new proteins in the latex of this plant. Quantitative analysis by normalized spectral counting revealed 10 down-regulated proteins in the latex of diseased plants, 9 cysteine proteases (chymopapain) and 1 latex serine proteinase inhibitor. A repression of papaya latex proteolytic activity during PMeV infection was hypothesized. This was further confirmed by enzymatic assays that showed a reduction of cysteine-protease-associated proteolytic activity in the diseased papaya latex. These findings are discussed in the context of plant responses against pathogens and may greatly contribute to understand the roles of laticifers in plant stress responses.

  12. Autografting with CD34+ peripheral blood stem cells: retained engraftment capability and reduced tumour cell content. (United States)

    Voso, M T; Hohaus, S; Moos, M; Pförsich, M; Cremer, F W; Schlenk, R F; Martin, S; Hegenbart, U; Goldschmidt, H; Haas, R


    The efficacy of an immunomagnetic purging method and the Isolex 300 devices were assessed for selecting CD34+ cells from leukapheresis products of 29 patients with non-Hodgkin's lymphoma (NHL), 39 with multiple myeloma and 34 with breast cancer. The mean purity of the CD34+ cell population was 93.6% and the mean recovery was 67.7%. Following enzymatic cleavage by chymopapain the expression of Thy-1 and Leu-8 was significantly reduced without affecting haematological recovery. The population of selected CD34+ cells of 4/8 patients with follicular lymphoma became PCR-negative. A 2.5 log reduction of tumour cells could be achieved in four patients with multiple myeloma as shown by a quantitative PCR assay. There were no tumour cells detectable in any of the 19 CD34+ cell preparations of patients with breast cancer. In 64 patients who received 94 cycles of high-dose therapy, a mean number of 4.7x 10(6) CD34+ cells/kg were autografted. The time needed for platelet reconstitution was different when a comparison was made with 156 patients, who had received unmanipulated leukapheresis products (10 v 12 d, P = 0.006). No significant differences with regard to neutrophil recovery were noted. Five patients had a graft failure. Two of them died (on day 78 and 88 following PBSCT), and three patients were rescued with unmanipulated back-up transplants. In conclusion, the immunomagnetic selection of CD34+ cells provides autografts with reduced tumour cell content and an engraftment ability similar to that of unmanipulated autografts.

  13. Papain Induced Occupational Asthma with Kiwi and Fig Allergy. (United States)

    Jiang, Nannan; Yin, Jia; Wen, Liping


    Papain is a proteolytic enzyme which is widely used in food industry, pharmaceuticals, and cosmetics. Occupational and non-occupational papain allergies have previously been documented; however, there are limited publications about papain allergy with its relative fruit allergy. Here, we present a case of occupational, IgE-mediated papain allergy with kiwi fruit and fig fruit allergy. A 53-year-old man suffered from rhinitis for several years, with the onset of his symptoms coinciding with the time he started to work at a sausage processing plant where papain is often used as a meat tenderizer. He began to experience symptoms of chest tightness, shortness of breath and wheezing shortly after starting work 5 years ago. Furthermore, he experienced several episodes of oral itching, and tongue and oropharyngeal angioedema after injestion of kiwi fruit and fig fruit. The patient had a lifelong history of allergic conjunctivitis, allergic rhinitis, and childhood asthma. Specific IgE was positive to kiwi fruit, papain and chymopapain (2.95 kUA/L, >100 kUA/L, and 95.0 kUA/L, respectively). Similar bands at 10-15 kDa in blotting with papain and kiwi fruit extracts were found. This patient showed a potential association between papain allergy and sensitization to kiwi fruit. We also reviewed 13 patients with papain allergy published in the literature, with 85% (11/13) of the patients sensitized through the respiratory tract, and 40% (4/11) having atopy. Further studies should focus on the determination of cross-reactive allergens between papain and its fruit relatives, and the prevalence of food allergy in patients with papain allergy should be investigated in a relatively large cohort.

  14. Articular chondrocyte metabolism and osteoarthritis

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    Leipold, H.R.


    The three main objectives of this study were: (1) to determine if depletion of proteoglycans from the cartilage matrix that occurs during osteoarthritis causes a measurable increase of cartilage proteoglycan components in the synovial fluid and sera, (2) to observe what effect intracellular cAMP has on the expression of matrix components by chondrocytes, and (3) to determine if freshly isolated chondrocytes contain detectable levels of mRNA for fibronectin. Canine serum keratan sulfate and hyaluronate were measured to determine if there was an elevation of these serum glycosaminoglycans in a canine model of osteoarthritis. A single intra-articular injection of chymopapain into a shoulder joint increased serum keratan sulfate 10 fold and hyaluronate less than 2 fold in 24 hours. Keratan sulfate concentrations in synovial fluids of dogs about one year old were unrelated to the presence of spontaneous cartilage degeneration in the joints. High keratan sulfate in synovial fluids correlated with higher keratan sulfate in serum. The mean keratan sulfate concentration in sera of older dogs with osteoarthritis was 37% higher than disease-free controls, but the difference between the groups was not statistically significant. Treatment of chondrocytes with 0.5 millimolar (mM) dibutyryl cAMP (DBcAMP) caused the cells to adopt a more rounded morphology. There was no difference between the amount of proteins synthesized by cultures treated with DBcAMP and controls. The amount of fibronectin (FN) in the media of DBcAMP treated cultures detected by an ELISA was specifically reduced, and the amount of {sup 35}S-FN purified by gelatin affinity chromatography decreased. Moreover, the percentage of FN containing the extra domain. A sequence was reduced. Concomitant with the decrease in FN there was an increase in the concentration of keratan sulfate.

  15. A history of lumbar disc herniation from Hippocrates to the 1990s. (United States)

    Truumees, Eeric


    In ancient times, a supernatural understanding of the syndrome of lumbar radiculopathy often involved demonic forces vexing the individual with often crippling pain. The ancient Greeks and Egyptians began to take a more naturalistic view and, critically, suspected a relationship between lumbar spinal pathology and leg symptoms. Relatively little then changed for those with sciatica until the classic works by Cotugno and Kocher arrived in the late 18th century. Early lumbar canal explorations were performed in the late 1800s and early 1900s by MacEwen, Horsley, Krause, Taylor, Dandy, and Cushing, among others. In these cases, when compressive pathologies were found and removed, the lesions typically were (mis-)identified as enchondromas or osteochondritis dissecans. To better understand the history, learn more about the first treatments of lumbar disc herniation, and evaluate the impact of the early influences on modern spine practice, searches of PubMed and Embase were performed using the search terms discectomy, medical history, lumbar spine surgery, herniated disc, herniated nucleus pulposus, sciatica, and lumbar radiculopathy. Additional sources were identified from the reference lists of the reviewed papers. Many older and ancient sources including De Ischiade Nervosa are available in English translations and were used. When full texts were not available, English abstracts were used. The first true, intentional discectomy surgery was performed by Mixter and Barr in 1932. Early on, a transdural approach was favored. In 1938, Love described the intralaminar, extradural approach. His technique, although modified with improved lighting, magnification, and retractors, remains a staple approach to disc herniations today. Other modalities such as chymopapain have been investigated. Some remain a part of the therapeutic armamentarium, whereas others have disappeared. By the 1970s, CT scanning after myelography markedly improved the clinical evaluation of patients with

  16. Quimionucleólise cervical associada à espondilectomia ventral em cães: aspectos clínico-cirúrgicos, radiográficos e histológicos Cervical chemonucleolysis associated to ventral slot in dogs: clinical-surgical, radiological, and histological aspects

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    A.P.F. Daibert


    Full Text Available Foram avaliados aspectos clínico-cirúrgicos, radiográficos e histológicos de discos intervertebrais cervicais caninos após quimionucleólise com quimiopapaína associada à espondilectomia ventral em 24 cães. Em todos foi realizada a espondilectomia ventral na região do disco intervertebral C2-C3 e quimionucleólises nos demais discos cervicais. Os cães foram sacrificados às 24 horas, e aos 8, 30, 60, 90 e 120 dias após aplicação da enzima para a obtenção de cortes sagitais dos discos intervertebrais tratados e das estruturas cartilaginosas e ósseas adjacentes. Nos mesmos períodos, foram realizadas radiografias para monitoração da espessura dos espaços intervertebrais. Todos toleraram bem o procedimento cirúrgico e não apresentaram alterações clínicas significativas. Na avaliação radiográfica, observou-se marcada redução dos espaços intervertebrais 24 horas após o tratamento. Do oitavo ao 90º dia após aplicação da enzima, foi verificada ausência total de espaço intervertebral na região dos discos tratados. Aos 120 dias, as espessuras dos espaços intervertebrais corresponderam, em média, a 59,1% da espessura anterior ao procedimento. Na avaliação histológica, foram observadas digestão nuclear e redução da intensidade de coloração pela safranina-O 24 horas após a quimionucleólise. A partir do 60º dia, havia material amorfo no espaço nuclear, que, aos 120 dias, tinha aparência fibrocartilaginosa. A quimionucleólise associada à espondilectomia ventral determinou a lise dos discos intervertebrais e lesões nas estruturas cartilaginosas e ósseas adjacentes, com tendência à reparação por tecido fibrocartilaginoso ao longo do tempo.The clinical-surgical, radiological, and histological aspects of chemonucleolysis with chymopapain in canine cervical intervertebral disks associated to ventral slot in 24 dogs were evaluated. The ventral slot was accomplished in the area of the intervertebral disks


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    Sakuntala Putrevu


    Full Text Available BACKGROUND Thrombocytopenia is defined as platelet count less than one and half lakh per mm cube of blood. Thrombocytopenia has been documented in patients with infectious mononucleosis, cytomegalovirus, and varicella zoster infections. Hepatitis C, tuberculosis and human immunodeficiency virus also have been reported in the causes list. Thrombocytopenia is a well-known complication of chronic lymphocytic leukaemia, although it may not be encountered as easily as seen in autoimmune haemolytic anaemias in these patients. It has also been reported in patients with other lymphoproliferative disorders including Hodgkin’s disease. Thrombocytopenia in patients with a variety of solid tumours has also been thought to most likely be immune mediated. Thrombocytopenia may accompany Graves’ disease and Hashimoto’s thyroiditis, but it is not certain that it is immunologically mediated or not. In our country, Dengue and malaria remain the predominant cause for Thrombocytopenia. Chymopapain and papain are the two important compound that are present in papaya. Lipase, a hydrolase, which is tightly bonded to the waterinsoluble fraction of crude papain is also seen in papaya. The papaya seeds and fruits have excellent antibiotic properties. The consumption of unripe and semi-ripe papaya fruits could be dangerous during pregnancy as it contains papain which may cause natural uterine contraction and may lead to abortions. It is reported that the plant extracts of papaya have maximum activity against dengue virus. It also has been reported that the methyl gallate of plant origin interacts with herpes simplex virus and causes its destruction. Majority of the papaya plant parts is known to have antimicrobial property. This study puts in a sincere effort to check the effects of papaya extract in the treatment of low platelet count. This study is intended to help the practising physicians to understand the benefits of the use of papaya extract when using in the