Interplay Between the IL-33/ST2 Axis and Bone Marrow ILC2s in Protease Allergen-Induced IL-5-Dependent Eosinophilia.

Background: Eosinophils develop from CD34+ progenitor cells in the bone marrow under the influence of interleukin (IL)-5. Several cell types produce IL-5, including type 2 innate lymphoid cells (ILC2s). The alarmin cytokine IL-33 is known to activate ILC2s in mucosal tissues, but little is known about IL-33-responsive ILC2s in the bone marrow in allergen-induced airway inflammation. Methods: Wild type (WT) and Rag1 deficient (Rag1-/-) mice, which lack mature T and B cells, received intranasal doses of papain to induce acute allergic inflammation. In some experiments, mice were pre-treated with anti-IL-5 prior to the papain challenge. Furthermore, recombinant IL-33 was administered to WT mice, Rag1-/- mice, lymphocyte deficient mice (Rag2-/-Il2rg-/-) and to ex vivo whole bone marrow cultures. Bone marrow eosinophils and ILC2s were analyzed by flow cytometry. Eosinophil count was assessed by differential cell count and secreted IL-5 from bone marrow cells by ELISA. Results: Intranasal administration of papain or IL-33 increased the number of mature eosinophils in the bone marrow despite the absence of adaptive immune cells in Rag1-/- mice. In parallel, an increased number of eosinophils was observed in the airways together with elevated levels of Eotaxin-2/CCL24. Bone marrow ILC2s were increased after papain or IL-33 administration, whereas ILC2s was found to be increased at baseline in Rag1-/- mice compared to WT mice. An upregulation of the IL-33 receptor (ST2) expression on bone marrow ILC2s was observed after papain challenge in both Rag1-/- and WT mice which correlated to increased number of bone marrow eosinophilia. Furthermore, an increased number of ST2+ mature eosinophils in the bone marrow was observed after papain challenge, which was further dependent on IL-5. In addition, bone marrow-derived ILC2s from both mouse strains produced large amounts of IL-5 ex vivo after IL-33 stimulation of whole bone marrow cultures. In contrast, IL-33-induced bone marrow and airway eosinophilia were abolished in the absence of ILC2s in Rag2-/-Il2rg-/- mice and no production of IL-5 was detected in IL-33-stimulated bone marrow cultures. Conclusion: These findings establish bone marrow ILC2s and the IL-33/ST2 axis as promising targets for modulation of uncontrolled IL-5-dependent eosinophilic diseases including asthma.

Pulmonary Emphysema Impairs Male Reproductive Physiology Due To Testosterone and Oxidative Stress Imbalance in Mesocricetus auratus.

This study evaluated whether pulmonary emphysema affects sperm quality, male reproductive organs, and testosterone levels in adult male hamsters. Mesocricetus auratus males (130-150 g) were subdivided into a control group (C group) and an emphysema group (E group). The C group received an intratracheal instillation of saline solution (0.3 mL/100 g of body weight), and the E group received papain (40 mg/100 g of body weight). After 60 days, the biometric, pulmonary, and reproductive parameters of each group were evaluated. The E group developed pulmonary emphysema, which decreased body weight and sperm quality compared to the C group. In oxidative stress-related assays, lipid peroxidation was increased in the testis and epididymis (caput and cauda) in the E group compared with the C group. However, only the caput epididymis showed a reduction in glutathione levels. Pulmonary emphysema also affected the testicle by inducing an increase in abnormal seminiferous tubules, accompanied by a decrease in seminiferous epithelium height. Spermatogenesis kinetics were also modified by pulmonary emphysema. The number of Leydig and Sertoli cells decreased in the E group, accompanied by an increase in the nuclear volume of Leydig cells. Testosterone concentration was increased in the E group. Similarly, pulmonary emphysema altered epididymal components in all regions. In conclusion, pulmonary emphysema affected the reproductive system in this experimental model, as shown by testicular and epididymal morphophysiology changes, hormonal alteration, and oxidative stress imbalance, inducing the loss of correct function in the male reproductive system.

Papain grafted into the silica coated iron-based magnetic nanoparticles 'IONPs@SiO2-PPN' as a new delivery vehicle to the HeLa cells.

The present study aims at engineering, fabrication, characterization, and qualifications of papain (PPN) conjugated SiO2-coated iron oxide nanoparticles 'IONPs@SiO2-PPN'. Initially fabricated iron oxide nanoparticles (IONPs) were coated with silica (SiO2) using sol-gel method to hinder the aggregation and to enhance biocompatibility. Next, PPN was loaded as an anticancer agent into the silica coated IONPs (IONPs@SiO2) for the delivery of papain to the HeLa cancer cells. This fabricated silica-coated based magnetic nanoparticle is introduced as a new physiologically-compatible and stable drug delivery vehicle for delivering of PPN to the HeLa cancer cell line. The IONPs@SiO2-PPN were characterized using FT-IR, AAS, FESEM, XRD, DLS, and VSM equipment. Silica was amended on the surface of iron oxide nanoparticles (IONPs, γ-Fe2O3) to modify its biocompatibility and stability. The solvent evaporation method was used to activate PPN vectorization. The following tests were performed to highlight the compatibility of our proposed delivery vehicle: in vitro toxicity assay, in vivo acute systemic toxicity test, and the histology examination. The results demonstrated that IONPs@SiO2-PPN successfully reduced the IC50 values compared with the native PPN. Also, the structural alternations of HeLa cells exposed to IONPs@SiO2-PPN exhibited higher typical hallmarks of apoptosis compared to the cells treated with the native PPN. The in vivo acute toxicity test indicated no clinical signs of distress/discomfort or weight loss in Balb/C mice a week after the intravenous injection of IONPs@SiO2 (10 mg kg-1). Besides, the tissues architectures were not affected and the pathological inflammatory alternations detection failed. In conclusion, IONPs@SiO2-PPN can be chosen as a potent candidate for further medical applications in the future, for instance as a drug delivery vehicle or hyperthermia agent.

Plant latex thrombin-like cysteine proteases alleviates bleeding by bypassing factor VIII in murine model.

Hemostasis is a tightly regulated process which maintains a fluid state of blood within the vasculature and provides thrombotic response upon tissue injury. Various scientific studies have implicated the role of plant latex proteases in hemostasis using in vitro experiments. However, in vivo models substantiate their role in hemostasis. Therefore, in the present study, the effect of plant latex thrombin-like proteases (PTLPs) on hemostasis was investigated systematically using mice tail bleeding as a preclinical model. In this direction, latex protease fractions (LPFs), which showed potent thrombin-like activity, were selected as they act directly on fibrinogen to form clot and quickly stop bleeding. Thrombin-like activity was exhibited mainly by cysteine proteases. Calotropis gigantea, Carica papaya, Jatropha curcas, Oxystelma esculentum, Tabernaemontana divaricata, and Vallaris solanacea LPFs and papain from C. papaya latex significantly reduced bleeding on a topical application in normal and aspirin administered mice. In addition, PTLPs accelerated the clotting of factor VIII deficient plasma, while, papain brought back the clotting time to normal levels acting like a bypassing agent. Further, papain failed to show activity in the presence of specific cysteine protease inhibitor iodoacetic acid; confirming protease role in all the activities exhibited. At the tested dose, PTLPs except C. gigantea did not show toxicity. Further, structural and sequence comparison between PTLPs and human thrombin revealed structural and sequence dissimilarity indicating their unique nature. The findings of the present study may open up a new avenue for considering PTLPs including papain in the treatment of bleeding wounds.

Runx/Cbfß complexes protect group 2 innate lymphoid cells from exhausted-like hyporesponsiveness during allergic airway inflammation.

Group 2 innate lymphoid cells (ILC2s) have tissue-resident competence and contribute to the pathogenesis of allergic diseases. However, the mechanisms regulating prolonged ILC2-mediated TH2 cytokine production under chronic inflammatory conditions are unclear. Here we show that, at homeostasis, Runx deficiency induces excessive ILC2 activation due to overly active GATA-3 functions. By contrast, during allergic inflammation, the absence of Runx impairs the ability of ILC2s to proliferate and produce effector TH2 cytokines and chemokines. Instead, functional deletion of Runx induces the expression of exhaustion markers, such as IL-10 and TIGIT, on ILC2s. Finally, these 'exhausted-like' ILC2s are unable to induce type 2 immune responses to repeated allergen exposures. Thus, Runx confers competence for sustained ILC2 activity at the mucosa, and contributes to allergic pathogenesis.

Protease loaded permeation enhancer liposomes for treatment of skin fibrosis arisen from second degree burn.

Cysteine protease (papain) is a plant derived enzyme and due to its collagenolytic activity has potential in fibrosis reduction. However, a major hurdle in its use as fibrosis reducing agent is to overcome stratum corneum skin barrier via topical application, owing to its hydrophilic and high molecular weight and protein nature which is prone to degradation. The aim of the present study was to develop a penetration enhancer incorporated drug delivery system, i.e. propylene glycol (PG) liposomes, loaded with papain for application in fibrosis therapy. Papain loaded PG-liposomes were prepared by the solvent injection method and characterized by size, shape, zeta potential, entrapment efficiency, drug release and stability. Papain conformational changes due to process stress were evaluated by electrophoresis and fluorescence spectroscopy. Biological evaluation was carried out in rodents by skin irritation and percent fibrosis reduction assays following induction of fibrosis arisen due to controlled second degree burn. Papain loaded PG-liposomes had mean vesicle size 180±30.3, zeta potential -25±1, polydispersity index 0.181 and 85±4.3% entrapment efficiency. Cumulative drug release after 8h was found to be 74.26±3.0%. SDS-PAGE and fluorescence spectroscopic studies confirmed the stability of papain after incorporation in PG-liposomes. Fibrosis reduction studies in animal models revealed that PG-liposomes incorporated papain improved fibrosis reduction significantly in comparison to conventional liposomes and free papain solution (p <0.05). Data suggest that propylene glycol incorporated liposomal system enhances papain proteolytic and collagenolytic activity along with a reduction in skin irritancy via preventing direct contact of papain with skin, improves papain therapeutic fibrosis reduction potential, an approach that may provide an efficient alternative for protease mediated fibrosis reduction in a variety of demanding circumstances.

[A 67-year old man with epigastric pain]. / 67-jähriger Patient mit epigastrischen Schmerzen.

A 67-year-old man suffering from epigastric pain showed a phytobezoar in the endoscopy. Therapy with Coca Cola® and enzymes was initiated. The (partial) lysis led to a migration of the bezoar into the ileum, resulting in a small bowel obstruction. After removal of the remaining bezoar via ileotomy a secondary pneumatosis intestinalis occurred. As a rare finding the (phyto-)bezoar should be considered as a differential diagnosis of abdominal pain - especially considering the rising numbers of bariatric surgery, which is a potential risk factor. Furthermore, intestinal obstruction after migration has to be considered as a relevant complication of treatment.

Papain wound dressings obtained from poly(vinyl alcohol)/calcium alginate blends as new pharmaceutical dosage form: Preparation and preliminary evaluation.

Transparent, soft, flexible, mechanically resistant films, which are ideal for use as wound dressings were prepared in the presence of 2% papain, a proteolytic enzyme that can play a role in the chemical debridement of the skin and can accelerate the healing process. The films, based on poly(vinyl alcohol):calcium alginate blends with increasing concentrations of polysaccharide (10, 20, and 30% v/v), were obtained by casting method. FTIR and DSC analyses were performed to assess the composition and miscibility of blends. Mechanical properties such as tensile strength, elasticity modulus, and elongation at breakpoint were evaluated. The influence of different concentrations of calcium alginate on physical attributes of films like wettability, swelling capacity and mechanical properties was determined. The stability of papain in the films was assessed indirectly by hemolytic activity assay employing direct contact method and confirmed by technique based on blood agar diffusion. Preliminary cytotoxicity was evaluated with the XTT method. The results showed that at the polymer concentrations tested, the blends were miscible. The increase in the content of the calcium alginate increased the wettability and swelling capacity of the films, which is desirable in wound dressings. On the other hand, mechanical resistance decreased without causing breakage of the films during the swelling tests. The hemolytic activity of the films was maintained during the studied period, suggesting the stability of papain in the proposed formulations. Cellular viability indicated that the films were non-toxic. The analysis of the results showed that it is possible to prepare interactive and bioactive wound dressing containing papain from blends of PVA and calcium alginate polymers.

Effect of the addition of papain enzyme on digestive parameters and palatability of extruded diets for dogs / Efeito da adição da enzima papaína em rações extrusadas para cães sobre parâmetros digestivos e palatabilidade

Common protein sources used in the manufacturing of diets for dogs are derived from by-products, which may have reduced digestibility depending on the source. This study evaluated the effect of the addition of a protease, the papain enzyme, as a supplement to extruded diets on palatability, nutrient digestibility, and fecal production and quality of dogs. A diet was formulated with poultry by-product meal, meat and bone meal, and feather meal as protein sources. This formula was divided into three isonutrient diets: one negative control (NC), without enzymes; treatment one (EZ1) with addition of 855.000UI of papain per kilogram of diet, and treatment two (EZ2) with addition of 2.280.000UI of papain per kilogram of diet, both added before extrusion. The experiment followed a randomized block design, with two blocks of nine animals (three animals per treatment in each block), 18 dogs in total, and six replicates per treatment. Data were submitted to analysis of variance and the means of three treatments were compared by polynomial contrasts (P <0.05). No differences in the coefficients of total tract apparent digestibility of nutrients nor changes in palatability, pH, and fecal production among treatments were found with the addition of different doses of enzyme to the diets (P > 0.05). The fecal score was reduced with increased addition of enzyme (P < 0.05).(AU)

As fontes comuns de proteína utilizadas na fabricação de rações para cães são oriundas de coprodutos, os quais podem apresentar digestibilidade reduzida de acordo com a fonte. Este estudo avaliou os efeitos da adição da enzima papaína em dietas secas e extrusadas na palatabilidade, digestibilidade dos nutrientes, qualidade e produção fecal de cães adultos. Uma dieta foi formulada contendo farinha de vísceras de frango, farinha de carne e ossos e farinha de penas hidrolisadas como fontes proteicas. Esta foi posteriormente dividida em três dietas isonutrientes: controle negativo (CN) sem adição da enzima; adição de 855.000 UI de papaína por quilograma de ração (EZ1); e adição de 2.280.000 UI de papaína por quiilograma de ração (EZ2), ambas adições feitas antes da extrusão. O experimento seguiu delineamento em blocos casualizados, com dois blocos de nove animais (três animais por tratamento em cada bloco), totalizando 18 cães, e seis repetições por tratamento. Os dados obtidos foram submetidos a análise de variância, com as médias dos três tratamentos comparadas por contrastes polinomiais (P < 0,05). Não foram verificadas diferenças nos coeficientes de digestibilidade aparente dos nutrientes ou mesmo alterações na palatabilidade, pH e produção de fezes entre os tratamentos com diferentes inclusões de enzima (P > 0,05). Apenas o escore fecal reduziu com o aumento da adição da enzima (P < 0,05).(AU)

Papain-Based Vaccination Modulates Schistosoma mansoni Infection-Induced Cytokine Signals.

We have previously shown that immunization of outbred rodents with cysteine peptidases-based vaccine elicited type 2-biased immune responses associated with consistent and reproducible protection against challenge Schistosoma mansoni. We herein start to elucidate the molecular basis of C57BL/6 mouse resistance to S. mansoni following treatment with the cysteine peptidase, papain. We evaluated the early cytokine signals delivered by epidermal, dermal, and draining lymph node cells of naïve, and S. mansoni -infected mice treated 1 day earlier with 0 or 50 µg papain, or immunized twice with papain only (10 µg/mouse), papain-free recombinant S. mansoni glyceraldehyde 3-phosphate dehydrogenase and 2-Cys peroxiredoxin peptide (10 and 15 µg/mouse, respectively = antigen Mix), or papain-adjuvanted antigen Mix. Schistosoma mansoni infection induced epidermal and lymph node cells to release type 1, type 2 and type 17 cytokines, known to counteract each other. Expectedly, humoral immune responses were negligible until patency. Papain pretreatment or papain-based vaccination diminished or shut off S. mansoni infection early induction of type 1, type 17 and type 2 cytokines except for thymic stromal lymphopoietin and programmed the immune system towards a polarized type 2 immune milieu, associated with highly significant (P < 0.005 - <0.0001) resistance to S. mansoni infection.

Intrinsic functional defects of type 2 innate lymphoid cells impair innate allergic inflammation in promyelocytic leukemia zinc finger (PLZF)-deficient mice.

BACKGROUND: The transcription factor promyelocytic leukemia zinc finger (PLZF) is transiently expressed during development of type 2 innate lymphoid cells (ILC2s) but is not present at the mature stage. We hypothesized that PLZF-deficient ILC2s have functional defects in the innate allergic response and represent a tool for studying innate immunity in a mouse with a functional adaptive immune response. OBJECTIVE: We determined the consequences of PLZF deficiency on ILC2 function in response to innate and adaptive immune stimuli by using PLZF(-/-) mice and mixed wild-type:PLZF(-/-) bone marrow chimeras. METHODS: PLZF(-/-) mice, wild-type littermates, or mixed bone marrow chimeras were treated with the protease allergen papain or the cytokines IL-25 and IL-33 or infected with the helminth Nippostrongylus brasiliensis to induce innate type 2 allergic responses. Mice were sensitized with intraperitoneal ovalbumin-alum, followed by intranasal challenge with ovalbumin alone, to induce adaptive TH2 responses. Lungs were analyzed for immune cell subsets, and alveolar lavage fluid was analyzed for ILC2-derived cytokines. In addition, ILC2s were stimulated ex vivo for their capacity to release type 2 cytokines. RESULTS: PLZF-deficient lung ILC2s exhibit a cell-intrinsic defect in the secretion of IL-5 and IL-13 in response to innate stimuli, resulting in defective recruitment of eosinophils and goblet cell hyperplasia. In contrast, the adaptive allergic inflammatory response to ovalbumin and alum was unimpaired. CONCLUSIONS: PLZF expression at the innate lymphoid cell precursor stage has a long-range effect on the functional properties of mature ILC2s and highlights the importance of these cells for innate allergic responses in otherwise immunocompetent mice.

Effectiveness Of 2% And 4% Papain Gels In The Healing Of Venous Ulcers / Efectividad De Los Geles De Papaína Al 2% Y Al 4% En La Cicatrización De Úlceras Venosas / Efetividade Dos Géis De Papaína A 2% E 4% Na Cicatrização De Úlceras Venosas

OBJECTIVE To analyze the effectiveness of 2% and 4% papain gels in tissue repair of venous ulcers. METHOD Quasi-experimental study with consecutive sample of 16 patients with 30 venous ulcers treated at the outpatient clinic of a teaching hospital, from April to November in 2011, using a form for clinical assessment of the patient and its lesion. Variables were analyzed by Wilcoxon and McNemar test (p < 0.05). RESULTS Most participants were female; aged between 51 and 59 years; obese; with hypertension. Regarding ulcers, there was an average decrease of 7.9 cm2 (50% of its original size) in 90 days; 20% of the ulcers completely healed within 56.67 days. There was an increase in epithelialization, significant reduction in the slough and edema, improved depth, in the type and amount of exudate (p < 0.0001). CONCLUSION 2% and 4% papain gels were effective in healing venous ulcers. .

OBJETIVO Analizar la efectividad de los geles de papaína al 2% y al 4% en la reparación del tejido de las úlceras venosas. MÉTODO Estudio cuasi-experimental, con muestra consecutiva de 16 pacientes con 30 úlceras venosas, atendidos en el ambulatorio de un hospital universitario, de abril a noviembre de 2011, con formulario para evaluación clínica del paciente y la lesión. Variables analizadas por las pruebas de Wilcoxon y McNemar (p < 0,05). RESULTADOS Predominio del sexo femenino; edad entre 51 y 59 años; obesas; con Hipertensión Arterial Sistémica. En cuanto a las úlceras, hubo reducción media de 7,9 cm2 (el 50% del tamaño) en 90 días; el 20% cicatrizaron completamente en 56,67 días. Hubo incremento de la epitelización, reducción significativa de la esfacelación y el edema, mejora en la profundidad, la clase y la cantidad de exudado (p < 0,0001). CONCLUSIÓN Los geles de papaína al 2% y al 4% fueron efectivos en la cicatrización de úlceras venosas. .

OBJETIVO Analisar a efetividade dos géis de papaína a 2% e 4% no reparo tecidual das úlceras venosas. MÉTODO Estudo quase-experimental, com amostra consecutiva de 16 pacientes com 30 úlceras venosas, atendidos no ambulatório de um hospital universitário, de abril a novembro de 2011, com formulário para avaliação clínica do paciente e da lesão. Variáveis analisadas pelos testes de Wilcoxon e McNemar (p < 0,05). RESULTADOS Predomínio do sexo feminino; idade entre 51 e 59 anos; obesas; com Hipertensão Arterial Sistêmica. Quanto às úlceras, houve redução média de 7,9 cm2 (50% do tamanho) em 90 dias; 20% cicatrizaram completamente em 56,67 dias. Houve aumento da epitelização, redução significativa do esfacelo e do edema, melhora na profundidade, no tipo e na quantidade de exsudato (p < 0,0001). CONCLUSÃO Os géis de papaína a 2% e 4% foram efetivos na cicatrização de úlceras venosas. .

B cells regulate CD4+ T cell responses to papain following B cell receptor-independent papain uptake.

Papain, a cysteine protease allergen with inherent adjuvant activity, induces potent IL-4 expression by T cells in the popliteal lymph nodes of mice following footpad immunization. In this study, we identify a novel, non-BCR-mediated capacity for B cells to rapidly bind and internalize papain. B cells subsequently regulate the adaptive immune response by enhancing ICOS expression on CD4(+) T cells and amplifying Th2 and follicular helper T cell induction. Ab blockade of ICOS ligand, expressed by popliteal lymph node B cells, but not dendritic cells, at the peak of the response inhibits IL-4 responses in wild-type mice but not B cell-deficient mice. Thus, B cells play a critical role in amplifying adjuvant-dependent Th2 polarization following noncanonical acquisition and internalization of the cysteine protease papain.

Epicutaneous administration of papain induces IgE and IgG responses in a cysteine protease activity-dependent manner.

BACKGROUND: Epicutaneous sensitization to allergens is important in the pathogenesis of not only skin inflammation such as atopic dermatitis but also "atopic march" in allergic diseases such as asthma and food allergies. We here examined antibody production and skin barrier dysfunction in mice epicutaneously administered papain, a plant-derived occupational allergen belonging to the same family of cysteine proteases as mite major group 1 allergens. METHODS: Papain and Staphylococcus aureus V8 protease were patched on the backs of hairless mice. Transepidermal water loss was measured to evaluate the skin barrier dysfunction caused by the proteases. Papain or that treated with an irreversible inhibitor specific to cysteine proteases, E64, was painted onto the ear lobes of mice of an inbred strain C57BL/6. Serum total IgE levels and papain-specific IgE and IgG antibodies were measured by ELISA. RESULTS: Papain and V8 protease patched on the backs of hairless mice caused skin barrier dysfunction and increased serum total IgE levels, and papain induced the production of papain-specific IgG1, IgG2a, and IgG2b. Papain painted onto the ear lobes of C57BL/6 mice induced papain-specific IgE, IgG1, IgG2c, and IgG2b, whereas papain treated with E64 did not. IgG1 was the most significantly induced papain-specific IgG subclass among those measured. CONCLUSIONS: We demonstrated that the epicutaneous administration of protease not only disrupted skin barrier function, but also induced IgE and IgG responses in a manner dependent on its protease activity. These results suggest that protease activity contained in environmental sources contributes to sensitization through an epicutaneous route.

Complementary medicine on side-effects of adjuvant hormone therapy in patients with breast cancer.

BACKGROUND: This clinical investigation was performed in order to evaluate the benefit of complementary medicine in patients with breast cancer undergoing adjuvant hormone therapy (HT). PATIENTS AND METHODS: The patients (n=680) were treated according to international guidelines. All patients suffered from arthralgia and mucosal dryness induced by the adjuvant HT. In order to reduce side-effects, the patients were complementarily treated with a combination of sodium selenite, proteolytic plant enzymes (bromelaine and papain) and Lens culinaris lectin. On case report formulas, self assessment of defined side-effects of HT (namely arthralgia and mucosal dryness) were documented before and four weeks after complementary treatment. Validation was carried out by scoring from 1 (no side-effects/optimal tolerability) to 6 (extreme side-effects/extremely poor tolerability), however, only patients suffering from severe side-effects (symptom scores >3) were enrolled in this investigation. RESULTS: A total of 64% (316 out of 494) of patients suffering from severe arthralgia and 62% of patients (194 out of 310) with severe mucosal dryness significantly benefited from complementary medicine. The severity of side-effects of HT was reduced by complementary treatment. Mean scores of symptoms declined from 4.92 before treatment to 3.16 after four weeks of treatment for arthralgia and from 4.83 before treatment to 3.21 after four weeks of treatment for mucosal dryness, and these were the primary aims of this investigation. The reduction of side-effects of HT was statistically significant (p<0.001) after four weeks. CONCLUSION: This investigation further demonstrates benefits of indication-based complementary treatment with the combination of sodium selenite, proteolytic enzymes and L. culinaris lectin in patients with breast cancer.

Involvement of nuclear factor of activated T cells in granulocyte-macrophage colony-stimulating factor production in canine keratinocytes stimulated with a cysteine protease.

BACKGROUND: A previous study demonstrated that the cysteine protease of Dermatophagoides farinae induced production of granulocyte-macrophage colony-stimulating factor (GM-CSF) in a canine epidermal keratinocyte progenitor cell line (CPEK); however, the molecular mechanism has not been elucidated. HYPOTHESIS/OBJECTIVES: Given that the transcription of GM-CSF mRNA in human lymphocytes is mainly regulated by the nuclear factor of activated T cells (NFAT), it is hypothesized that NFAT also contributes to GM-CSF production in canine keratinocytes stimulated with a cysteine protease. METHODS: Nuclear translocation of NFAT was evaluated in CPEK cells in the absence or presence of the cysteine protease papain. We also investigated whether blockade of NFAT could inhibit GM-CSF production. RESULTS: Papain-induced nuclear translocation of NFAT, producing GM-CSF, was partly inhibited by ciclosporin. CONCLUSIONS AND CLINICAL IMPORTANCE: The results suggest that GM-CSF production mediated by the cysteine protease is regulated not only by NFAT but also by unknown signalling pathways in canine keratinocytes.

[Effects of proteolytic enzyme therapy with Wobe Mugos against chemotherapy-induced toxicity in breast cancer patients - results of a pilot study]. / Proteolytische enzymtherapie mit wobe mugos gegen chemotherapie-bedingte toxizitäten bei brustkrebs-patientinnen - ergebnisse einer pilotstudie.

BACKGROUND: Wobe Mugos(®) is an enzyme preparation containing the proteases trypsin and papain from the pancreatic calf and commonly used in complementary medicine. From non-randomized studies, its multiple favorable effects including the reduction of adverse events from radiotherapy and chemotherapy in oncology patients have been reported. METHODS: Patients with invasive breast cancer receiving adjuvant or palliative chemotherapy between 2005 and 2006 and who were scheduled for at least two further cycles of this specific chemotherapy were included in this pilot study. A specific toxicity of at least grade 2 using the NCI common toxicity criteria which occurred during the preceeding cycle and was relevant to the patient was recorded. This specific toxicity, e.g. grade 2 emesis, was again evaluated after two analogously administered further chemotherapy cycles in which Wobe Mugos(®) had been coadministered. The hypothesis was that specific toxicites of individual patients will be reduced by this enzyme therapy. The majority of the 57 consecutive patients received palliative chemotherapy. Peroral enzyme therapy was coadministered with two uncracked coated tablets three times daily on all days of a chemotherapy cycle except on the day of chemotherapy administration. RESULTS: Tolerability was good. Positive and neutral effects on toxicity parameters were observed in 11 and 42 patients, respectively, and a negative influence in 4 women. CONCLUSION: We observed only a marginal influence of Wobe Mugos(®) in patients with breast cancer who had experienced at least a grade 2 toxicity in the preceding cycle and who received two further identical cycles of this chemotherapy in conjunction with the enzyme preparation. Randomized studies on homogenous patient populations are necessary.

Mesenchymal stem cell transplantation increases expression of vascular endothelial growth factor in papain-induced emphysematous lungs and inhibits apoptosis of lung cells.

BACKGROUND: Pulmonary emphysema is characterized by loss of alveolar structures. We have found that bone marrow (BM) mesenchymal stem cell (MSC) transplantation ameliorates papain-induced pulmonary emphysema. However, the underlying mechanism is not completely understood. It has been shown that blocking the vascular endothelial growth factor (VEGF) signaling pathway leads to apoptosis of lung cells and pulmonary emphysema, and MSC are capable of secreting VEGF. We hypothesized that MSC transplantation may have a protective effect on pulmonary emphysema by increasing VEGF-A expression and inhibiting apoptosis of lung cells. METHODS: We examined the morphology and expression of VEGF-A in rat lung after papain treatment and MSC transplantation. We also used a co-culture system in which MSC and cells prepared from papain-treated lungs or control lungs were cultured together. The levels of VEGF-A in cells and culture medium were determined, and apoptosis of cultured lung cells was evaluated. RESULTS: VEGF-A expression in rat lungs was decreased after papain treatment, which was partly rescued by MSC transplantation. MSC production of VEGF-A was increased when MSC were co-cultured with cells prepared from papain-treated lungs. Furthermore, the apoptosis of papain-treated lung cells was inhibited when co-cultured with MSC. The induction of MSC production of VEGF-A by papain-treated lung cells was inhibited by adding anti-tumor necrosis factor (TNF)-alpha antibody to the medium. CONCLUSIONS: The protective effect of MSC transplantation on pulmonary emphysema may be partly mediated by increasing VEGF-A expression and inhibiting the apoptosis of lung cells. TNF-alpha released from papain-treated lung cells induces MSC to secret VEGF-A.

Study on the debridement efficacy of formulated enzymatic wound debriding agents by in vitro assessment using artificial wound eschar and by an in vivo pig model.

An in vitro efficacy study using newly developed artificial wound eschar (AWE) substrate was conducted for assessing enzyme dose response. The AWE substrate is prepared by the enzymatic conversion of fibrinogen to fibrin in the presence of collagen, fibrin, and elastin to form an insoluble planar matrix. AWE substrate was placed on Franz Diffusion Cells for continuously monitoring the debridement progress. A parallel in vivo study was performed using pig thermal-burn wounds. Papain at concentrations of 200, 400, 800, and 1,600 U/mg was used as the model debriding enzyme for both studies. The data from the first 5 hours of the in vitro testing showed that debriding activity increased as the enzyme concentration increased. The histological results of the in vivo biopsy samples showed that enzyme doses above 800 and 1,600 U/mg successfully achieved debridement on day 8, while lower treatment groups still contained eschar tissue. Using the histological measurement results (wound depth score) a dose response that correlated to the in vitro assessment was found. Granulation tissue maturity and reepithelialization displayed correlation with the enzyme dose. Results indicate that AWE substrate can be used to predict debridement efficacy in vitro when correlation to the in vivo assessment is achieved.

Bezoar gástrico como complicación de banda gástrica en manejo de obesidad mórbida: caso clínico / Gastric bezoar as complication of gastric banding: report of one case

Laparoscopic adjustable gastric banding (LAGB) is used for the management of morbid obesity. Phytobezoars are rarely reported as a complication of this operation and are usually extracted by endoscopic means. We report a 48 year-old male subjected to a gastric banding, that consulted for progressive dysphagia, six months after the operation. A barium meal x-ray examination demonstrated the presence of a bezoar that was dissolved in one week using papain. A control barium meal confirmed the disappearance of the bezoar.