Fast screening using attenuated total reflectance- fourier transform infrared (ATR-FTIR) spectroscopy of patients based on D-dimer threshold value.

Attenuated Total Reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy is an emerging technology in the medical field. Blood D-dimer was initially studied as a marker of the activation of coagulation and fibrinolysis. It is mainly used as a potential diagnosis screening test for pulmonary embolism or deep vein thrombosis but was recently associated with COVID-19 severity. This study aimed to evaluate the use of ATR-FTIR spectroscopy with machine learning to classify plasma D-dimer concentrations. The plasma ATR-FTIR spectra from 100 patients were studied through principal component analysis (PCA) and two supervised approaches: genetic algorithm with linear discriminant analysis (GA-LDA) and partial least squares with linear discriminant (PLS-DA). The spectra were truncated to the fingerprint region (1800-1000 cm-1). The GA-LDA method effectively classified patients according to D-dimer cutoff (≤0.5 µg/mL and >0.5 µg/mL) with 87.5 % specificity and 100 % sensitivity on the training set, and 85.7 % specificity, and 95.6 % sensitivity on the test set. Thus, we demonstrate that ATR-FTIR spectroscopy might be an important additional tool for classifying patients according to D-dimer values. ATR-FTIR spectral analyses associated with clinical evidence can contribute to a faster and more accurate medical diagnosis, reduce patient morbidity, and save resources and demand for professionals.

Anti-Fibronectin Aptamer Modifies Blood Clot Pattern and Stimulates Osteogenesis: An Ex Vivo Study.

BACKGROUND: Scaffold (SCA) functionalization with aptamers (APT) provides adsorption of specific bioactive molecules on biomaterial surfaces. The aim of this study was to observe if SCA enriched with anti-fibronectin APT can favor coagulum (PhC) and osteoblasts (OSB) differentiation. METHODS: 20 µg of APT was functionalized on SCA by simple adsorption. For PhC formation, SCAs were inserted into rat calvaria defects for 17 h. Following proper transportation (buffer solution PB), OSBs (UMR-106 lineage) were seeded over PhC + SCAs with and without APT. Cells and PhC morphology, PhC cell population, protein labeling and gene expression were observed in different time points. RESULTS: The APT induced higher alkaline phosphatase and bone sialoprotein immunolabeling in OSB. Mesenchymal stem cells, leukocytes and lymphocytes cells were detected more in the APT group than when scaffolds were not functionalized. Additionally, an enriched and dense fibrin network and different cell types were observed, with more OSB and white blood cells in PhC formed on SCA with APT. The gene expression showed higher transforming growth factor beta 1 (TGF-b1) detection in SCA with APT. CONCLUSIONS: The SCA functionalization with fibronectin aptamers may alter key morphological and functional features of blood clot formation, and provides a selective expression of proteins related to osteo differentiation. Additionally, aptamers increase TGF-b1 gene expression, which is highly associated with improvements in regenerative therapies.

Influence of a Physiologically Formed Blood Clot on Pre-Osteoblastic Cells Grown on a BMP-7-Coated Nanoporous Titanium Surface.

Titanium (Ti) nanotopography modulates the osteogenic response to exogenous bone morphogenetic protein 7 (BMP-7) in vitro, supporting enhanced alkaline phosphatase mRNA expression and activity, as well as higher osteopontin (OPN) mRNA and protein levels. As the biological effects of OPN protein are modulated by its proteolytic cleavage by serum proteases, this in vitro study evaluated the effects on osteogenic cells in the presence of a physiological blood clot previously formed on a BMP-7-coated nanostructured Ti surface obtained by chemical etching (Nano-Ti). Pre-osteoblastic MC3T3-E1 cells were cultured during 5 days on recombinant mouse (rm) BMP-7-coated Nano-Ti after it was implanted in adult female C57BI/6 mouse dorsal dermal tissue for 18 h. Nano-Ti without blood clot or with blood clot at time 0 were used as the controls. The presence of blood clots tended to inhibit the expression of key osteoblast markers, except for Opn, and rmBMP-7 functionalization resulted in a tendency towards relatively greater osteoblastic differentiation, which was corroborated by runt-related transcription factor 2 (RUNX2) amounts. Undetectable levels of OPN and phosphorylated suppressor of mothers against decapentaplegic (SMAD) 1/5/9 were noted in these groups, and the cleaved form of OPN was only detected in the blood clot immediately prior to cell plating. In conclusion, the strategy to mimic in vitro the initial interfacial in vivo events by forming a blood clot on a Ti nanoporous surface resulted in the inhibition of pre-osteoblastic differentiation, which was minimally reverted with an rmBMP-7 coating.

Old Methods, New Insights: Reviewing Concepts on the Ecology of Trypanosomatids and Bodo sp. by Improving Conventional Diagnostic Tools.

Mixed infections by different Trypanosoma species or genotypes are a common and puzzling phenomenon. Therefore, it is critical to refine the diagnostic techniques and to understand to what extent these methods detect trypanosomes. We aimed to develop an accessible strategy to enhance the sensitivity of the hemoculture, as well as to understand the limitations of the hemoculture and the blood clot as a source of parasitic DNA. We investigated trypanosomatid infections in 472 bats by molecular characterization (18S rDNA gene) of the DNA obtained from the blood clot and, innovatively, from three hemoculture sample types: the amplified flagellates ("isolate"), the pellet of the culture harvested in its very initial growth stage ("first aliquot"), and the pellet of non-grown cultures with failure of amplification ("sediment"). We compared (a) the characterization of the flagellates obtained by first aliquots and isolates; and (b) the performance of the hemoculture and blood clot for trypanosomatid detection. We observed: (i) a putative new species of Bodo in Artibeus lituratus; (ii) the potential of Trypanosoma cruzi selection in the hemoculture; (iii) that the first aliquots and sediments overcome the selective pressure of the hemoculture; and (iv) that the blood clot technique performs better than the hemoculture. However, combining these methods enhances the detection of single and mixed infections.

Elevación de seno maxilar sin injerto y colocación simultánea de implante dental / Graftless maxillary sinus lift with simultaneous dental implant placement

Introducción: La elevación de seno maxilar con implante dental simultáneo, sin el uso de substitutos óseos, ha retomado la teoría del potencial de neoformación ósea, que el coágulo de sangre alojado bajo la membrana de Schneider por sí solo presenta. Excluyendo así la necesidad de materiales exógenos. Objetivo: Exponer la efectividad de la elevación de seno maxilar con implante dental simultáneo, sin el uso de injerto, mediante la evaluación de la cantidad de ganancia ósea vertical. Presentación de casos: Caso 1: Paciente de 62 años, género femenino, que fue sometida a una elevación de seno maxilar sin injerto e implante dental simultáneo, con técnica de ventana lateral debido a altura ósea reducida de 5,24 mm. Luego del período de seguimiento tomográfico de 12 meses, después de la carga protésica, se logró una altura ósea vertical de 10,2 mm, lo que dio como resultado una ganancia ósea vertical de 4,96 mm. Caso 2: Paciente de 48 años, género femenino, que fue sometida a una elevación de seno maxilar sin injerto e implante dental simultáneo, con técnica transalveolar, mediante el uso de piezoeléctrico debido a una altura ósea reducida de 8,33 mm. Luego del período de seguimiento radiográfico de 4 meses, antes de la carga protésica, se logró una altura ósea vertical de 11,55 mm, lo que dio como resultado una ganancia ósea vertical de 3,19 mm. Conclusiones: Con base en estos 2 informes de casos, la elevación de seno maxilar e implante dental simultáneo sin injerto se asocian con la reducción de la morbilidad quirúrgica, menor probabilidad de procesos infecciosos y menor costo de la cirugía. Por lo tanto, puede considerarse una alternativa quirúrgica para la colocación de implantes en el maxilar posterior superior debido a deficiencias óseas verticales, independientemente de la técnica (lateral o transalveolar(AU)

Introduction: Maxillary sinus lift with simultaneous dental implantation without using bone substitutes, reapproaches the theory about the bone neoformation potential of the blood clot housed under the Schneider membrane, all by itself, thus excluding the need to use exogenous materials. Objective: Discuss the effectiveness of graftless maxillary sinus lift with simultaneous dental implant placement through an evaluation of the amount of vertical bone gain. Case presentation: Case 1: Female 62-year-old patient undergoing graftless maxillary sinus lift with simultaneous dental implantation by lateral window technique due to a reduced bone height of 5.24 mm. After a 12-month tomographic follow-up period subsequent to prosthetic loading, a 10.2 mm vertical bone height was achieved, resulting in 4.96 mm vertical bone gain. Case 2: Female 48-year-old patient undergoing graftless maxillary sinus lift with simultaneous dental implantation by transalveolar technique using a piezoelectric generator due to a reduced bone height of 8.33 mm. After a 4-month radiographic follow-up period, before prosthetic loading, an 11.55 mm vertical bone height was achieved, resulting in 3.19 mm vertical bone gain. Conclusions: According to these two case reports, graftless maxillary sinus lifting with simultaneous dental implantation is associated to reduced surgical morbidity, a lesser probability of infectious processes and lower surgical costs. It may therefore be considered to be a surgical alternative for implant placement in the posterior maxilla due to vertical bone deficiencies, regardless of the technique used (lateral or transalveolar(AU)

Optimal Parameters of Laser Therapy to Improve Critical Calvarial Defects.

Body bones play diverse pivotal roles, including the protection of vital organs. For instance, the integrative functions of the brain controlling diverse peripheral actions can be affected by a traumatic injury on the calvaria and the reparative process of a large defect is a challenge in the integrative physiology. Therefore, the development of biomaterials and approaches to improve such defects still requires substantial advances. In this regard, the most attractive approaches have been covering the cavity with inorganic bovine bone (IBB) and, more recently, also using low-level laser therapy (LT), but this issue has opened many questions. Here, it was determined the number of LT sessions required to speed up and to intensify the recovery process of two 5-mm-diameter defects promoted in the calvaria of each subgroup of six adult Wistar rats. The quantitative data showed that 30 days post-surgery, the recovery process by using blood clot-filling was not significantly influenced by the number of LT sessions. However, in the IBB-filled defects, the number of LT sessions markedly contributed to the improvement of the reparative process. Compared to the Control group (non-irradiated), the percentage of mineralization (formation of new bone into the cavities) gradually increased 25, 49, and 52% with, respectively, 4, 7, and 11 sessions of LT. In summary, combining the use of IBB with seven sessions of LT seems to be an optimal approach to greatly improve the recovery of calvarial defects. This translational research opens new avenues targeting better conditions of life for those suffering from large bone traumas and in the present field could contribute to preserve the integrative functions of the brain.

Trypanosoma spp. Neobats: Insights about those poorly known trypanosomatids.

Bats are infected with several trypanosomatid species; however, assessing the diversity of this interaction remains challenging since there are species apparently unable to grow in conventional culture media. Accordingly, the ecology and biology of the Molecular Operational Taxonomic Units (MOTUs) Trypanosoma spp. Neobats are unknown. Therefore, we performed the molecular characterization targeting the 18S small subunit rDNA from the blood clot of 280 bats of three Brazilian regions (Paraíba, Rio de Janeiro and Acre states), bypassing the selective pressure of hemoculture. From 68 (24%) positive blood clot samples, we obtained 49 satisfactory sequences. Of these successfully sequenced results, T. spp. Neobats (1, 3 and 4) represented 67%, with the most abundant T. sp. Neobat 4 (53%). Our results show: (1) high abundance and wide geographic range of T. sp. Neobat 4, restricted to Carollia bats; (2) high infection rate of T. sp. Neobat 4 in Carollia perspicillata populations (mean 26%); (3) infection with the monoxenous Crithidia mellificae; and (4) a new MOTU (T. sp. Neobat 5) in Artibeus cinereus, positioning in the Trypanosoma wauwau clade. These data corroborate the importance of bats as hosts of many Trypanosoma species and C. mellificae. They also show that the diversity of the T. wauwau clade is underestimated and warn about the high magnitude of trypanosomes we overpass with the hemoculture. Our findings combined with previous data show that T. spp. Neobats include host-specific and host-generalist species, probably playing different ecological roles: T. sp. Neobat 1 shows broad host range; T. spp. Neobat 3 and 4 are restricted to Artibeus and Carollia, respectively. Finally, T. Neobat 4 seems to be a well-succeeded parasite, especially within C. perspicillata metapopulations across a wide geographical distribution. This work is a step forward to understand the biology and life history of T. spp. Neobats.

Effect of ethylenediaminetetraacetic acid irrigation on immune-inflammatory response in teeth submitted to regenerative endodontic therapy.

AIM: To analyse longitudinally the immune-inflammatory response in teeth of mice that underwent a regenerative protocol with or without the use of ethylenediaminetetraacetic acid (EDTA) to irrigate the root canal system. METHODOLOGY: First maxillary molars of mice were devitalized using size 10 and 15 files. Teeth were divided into the following groups: Empty - the canals were left empty; Blood Clot (BC) - the canals were filled with a blood clot; and EDTA + Blood - the canals were irrigated with 0.06 mL of 17% EDTA for 1 min and filled with a blood clot. Access cavities were restored with Coltosol® . Animals were sacrificed at 7, 14 or 21 days after the operative procedures, and teeth were collected. RNA was extracted, mRNA expression of the cytokines IGF, NGF, IL-1α, IL-10, TGF and VEGF was assessed using real-time PCR, and the anova Kruskal-Wallis test was used. RESULTS: IL-1 mRNA expression was significantly higher in the EDTA + BC group than in the Empty and BC groups at the 7th and 14th days of evaluation (P < 0.05). IL-10 mRNA expression was similar across the three groups at all time periods. TGF-ß mRNA expression in the EDTA + BC group was significantly higher on the 7th and 21st days than on the 14th (P < 0.05); at day 21, TGF-ß mRNA expression was similar between the BC and EDTA + BC groups but significantly higher than in the Empty group (P < 0.05). IGF mRNA expression was significantly higher in the EDTA + BC group than in the other groups at all time periods. VEGF mRNA expression remained unchanged throughout the experimental period in all groups (P > 0.05). NGF mRNA expression was similar amongst all groups at the 7th and 21st days (P > 0.05). At the 14th day, however, there was a significant increase in NGF mRNA expression in the EDTA + Blood group (P < 0.05) when compared with the expression in the other groups. CONCLUSION: EDTA promoted increased expression of factors that have the potential to improve the outcome of regenerative endodontic treatment.

Uncovering Trypanosoma spp. diversity of wild mammals by the use of DNA from blood clots.

Trypanosoma spp. infection in wild mammals is detected mainly through parasitological tests that usually display low sensitivity. We propose the use of DNA extracted directly from blood clots (BC), which are neglected sources of DNA for diagnosis and identification of Trypanosoma spp. This approach followed by nested PCR targeting the 18S SSU rDNA demonstrated to be sensitive and suitable to evaluate the diversity of trypanosomes infecting sylvatic mammals, including subpatent and mixed infections. Infection was detected in 95/120 (79.2%) samples from bats, carnivores and marsupials that included negative serological and hemoculture testing mammals. Thirteen Trypanosoma spp. or Molecular Operational Taxonomic Units (MOTUs) were identified, including two new MOTUs. The high diversity of trypanosomes species and MOTUs infecting bats and marsupials showed that these hosts can be considered as bio-accumulators of Trypanosoma spp., with specimens of Didelphis spp. displaying the highest trypanosome diversity. The use of blood clots allowed direct access to non-culturable parasites, mixed infections, besides bypassing the selective pressure on the parasites inherent to cultivation procedures. Trypanosoma cruzi was the species found infecting the highest number of individuals, followed by T. lainsoni. Positive PCR for T. cruzi was observed in 16 seronegative individuals and 30 individuals with negative hemocultures. Also, T. lainsoni, previously found only in rodents, showed to be capable of infecting bats and marsupials. This finding makes it clear that some species of Trypanosoma are more generalist than previously thought. Molecular diagnosis using nested PCR from DNA extracted from BC allowed the increase of the knowledge about host-spectrum and distribution of Trypanosoma spp. and allowed the identification of new MOTUs.

Frozen blood clots can be used for the diagnosis of distinct Plasmodium species in man and non-human primates from the Brazilian Atlantic Forest.

BACKGROUND: Zoonotic infections with epidemic potential, as non-human primate malaria and yellow fever (YF), can overlap geographically. Optimizing a small blood sample for diagnosis and surveillance is of great importance. Blood are routinely collected for YF diagnosis and blood clots usually discarded after serum obtention. Aiming to take sample advantage, the sensitivity of a PCR using extracted DNA from long-term frozen clots from human and non-human primates for detection of Plasmodium spp. in low parasitaemia conditions was assayed. RESULTS: Malaria diagnosis with DNA extracted from blood clots generated results in agreement with samples obtained with whole blood, including mixed Plasmodium vivax/simium and Plasmodium malariae/brasilianum infections. CONCLUSION: Blood clots from human and non-human primates may be an important and low cost source of DNA for malaria surveillance in the Atlantic Forest.

Coagulation Profile Dynamics in Pediatric Patients with Cushing Syndrome: A Prospective, Observational Comparative Study.

OBJECTIVE: To evaluate the association between Cushing syndrome and hypercoagulability in children. STUDY DESIGN: A prospective, observational study was performed of 54 patients with Cushing syndrome, 15.1 ± 3.9 years, treated at the National Institutes of Health Clinical Center. Coagulation profiles were taken before and 6-12 months after surgery and compared with18 normocortisolemic children, 13.7 ± 3.6 years. RESULTS: At baseline, patients with Cushing syndrome had greater levels of the procoagulant factor VIII (FVIII) vs controls (145 IU/dL ± 84 vs 99 ± 47, P = .04); 6-12 months after surgery, FVIII levels decreased to 111 ± 47, P = .05. Patients with Cushing syndrome had greater levels of the antifibrinolytic α2-antiplasmin, 96 ± 17% vs 82 ± 26%, P = .015. After surgery, antifibrinolytic α2-antiplasmin levels decreased to 82 ± 24%, P < .001. Anticoagulants were greater in patients with Cushing syndrome vs controls at baseline, including protein C (138 ± 41% vs 84 ± 25%, P < .001), protein S (94 ± 19% vs 74 ± 19%, P = .001), and antithrombin III (96 ± 18% vs 77 ± 13%, P < .0001). The 24-hour urinary free cortisol levels correlated positively with FVIII levels, r = 0.43, P = .004. CONCLUSION: Children with Cushing syndrome had elevated procoagulants, antifibrinolytics, and anticoagulants at baseline compared with controls; normalization of coagulation measures was seen after surgical cure. Despite the increase in anticoagulants, hypercortisolemia is associated with a hypercoagulable state in children, as is the case in adults. This finding has potential implications for prevention of venous thromboembolism in children with Cushing syndrome. TRIAL REGISTRATION: ClinicalTrials.gov:NCT00001595.

Jejunal hemorrhage syndrome in a Zebu cow in Brazil / Síndrome de jejuno hemorrágico em gado Zebu no Brasil

Clostridium perfringens type A has been incriminated as the etiologic agent in jejunal hemorrhage syndrome (JHS), which is a disease that affects dairy cattle. Although this microorganism is considered an important enteropathogen the pathogenesis of JHS is still not clear, and there have been no reports of its occurrence in Brazil so far. The aim of this study was to describe the occurrence of JHS by infection with a C. perfringens type A strain carrying the beta-2 toxin gene in a zebu cow in Brazil, for the first time.

Clostridium perfringens tipo A tem sido considerado agente etiológico da síndrome do jejuno hemorrágico (SJH), que é uma doença que afeta comumente os rebanhos de gado. Embora este microrganismo seja considerado um importante enteropatógeno, a patogênese da SJH ainda não foi elucidada, e não havia sido reportada no Brasil até então. O alvo deste estudo foi descrever pela primeira vez a ocorrência da SJH causada por C. perfringens tipo A, carreador do gene da toxina beta-2, em um zebuíno no Brasil.

Jejunal hemorrhage syndrome in a Zebu cow in Brazil / Síndrome de jejuno hemorrágico em gado Zebu no Brasil

Clostridium perfringenstype A has been incriminated as the etiologic agent in jejunal hemorrhage syndrome (JHS), which is a disease that affects dairy cattle. Although this microorganism is considered an important enteropathogen the pathogenesis of JHS is still not clear, and there have been no reports of its occurrence in Brazil so far. The aim of this study was to describe the occurrence of JHS by infection with a C. perfringenstype A strain carrying the beta-2 toxin gene in a zebu cow in Brazil, for the first time.(AU)

Clostridium perfringens tipo A tem sido considerado agente etiológico da síndrome do jejuno hemorrágico (SJH), que é uma doença que afeta comumente os rebanhos de gado. Embora este microrganismo seja considerado um importante enteropatógeno, a patogênese da SJH ainda não foi elucidada, e não havia sido reportada no Brasil até então. O alvo deste estudo foi descrever pela primeira vez a ocorrência da SJH causadapor C. perfringenstipo A, carreador do gene da toxina beta-2, em um zebuíno no Brasil.(AU)

Maxillary sinus lift without grafting, and simultaneous implant placement: a prospective clinical study with a 51-month follow-up.

A prospective clinical study of maxillary sinus lift procedures in the posterior region of the maxilla, using only blood clot as filling material, was conducted. Seventeen patients underwent a maxillary sinus lift procedure; 20 maxillary sinus regions were operated on and a total of 25 implants were placed. The sinus mucosa was lifted together with the anterior wall of the osteotomized maxilla and supported by the implants placed. Computed tomography (CT) scans were obtained immediately postoperative (T initial) and at 3 (T1) and 51 (T2) months postoperative for the measurement of linear bone height and bone density (by grey tones). Only one implant was lost in the first stage (96% success). After dental prosthesis placement and during up to 51 months of follow-up, no implant was lost (100% success, second stage). The difference in mean bone height between T initial (5.94 mm) and T1 (13.14 mm), and between T initial and T2 (11.57 mm), was statistically significant (both P<0.001); comparison between T1 and T2 also presented a statistical difference (P<0.001). Bone density had increased at the end of the period analyzed, but this was not statistically significant (P>0.05). Thus, the maxillary sinus lift technique with immediate implant placement, filling with blood clot only, may be performed with a high success rate.

Revascularização pulpar: considerações técnicas e implicações clinícas / Pulp revascularization: technical considerations and clinical implications

O tratamento endodôntico de dentes portadores de necrose pulpar e rizogênese incompleta têm sido tratados, recentemente, por meio da revascularização pulpar. Essa promissora alternativa de tratamento promove tanto o fechamento apical quanto o término do desenvolvimento radicular. Atualmente vários protocolos têm sido propostos, entretanto pouco se sabe sobre o sucesso por meio da instrumentação mecânica e do uso de medicações a base de hidróxido de cálcio ou pasta tri-antibiótica nos casos com indicação de revascularização pulpar. Objetivo : O objetivo deste estudo foi realizar uma revisão da literatura abordando os protocolos de revascularização e suas implicações clínicas para o tratamento de dentes portadores de necrose pulpar e ápices incompletos. Resultados e discussão: As causas que normalmente interrompem a formação radicular são os traumatismos dentários e cáries dentárias, que podem causar a necrose pulpar. Sendo assim, os dentes que apresentam rizogênese incompleta e necrose pulpar geralmente eram tratados pelo método de apicificação ou mesmo a confecção de um plug apical de MTA, a fim de conseguir a formação de uma barreira apical. Porém, através desse método, as raízes continuam com as paredes dentinárias finas e fragilizadas. Conclusão: Com base nessa revisão, pode-se concluir que a revascularização pulpar é uma alternativa como tratamento para dentes com rizogênese incompleta portadores de necrose pulpar, porém, não há um protocolo estabelecido e considerado ideal...

Endodontic treatment of immature teeth with necrotic pulp and incomplete root formation has been recently treated with pulp revascularization. It is a promising alternative treatment to promote apical closure and root development. To date, a variety of revascularization protocols have been described, however little is known about the success of combining mechanical instrumentation and intracanal medication such as calcium hydroxide or triantibiotic paste. Thus, the aim of this study is to present a review of literature of pulp revascularization protocols and its clinical implications for treatment of teeth with pulp necrosis and incomplete apex. The causes that usually interrupt the root formation are dental traumatisms and caries, which can lead to pulp necrosis. Therefore, the immature permanent teeth and pulp necrosis were usually treated by apexification or the confection of an apical MTA plug, in order to accomplish the formation of an apical barrier. However, by this method, the roots canal walls remain thin and fragile. It may be concluded that the pulp revascularization treatment is an alternative approach for immature permanent teeth with pulp necrosis. However, there is not a standardized protocol that is considered ideal in these cases...

Potential application of protease isolated from Pseudomonas aeruginosa PD100

A protease isolated from Pseudomonas aeruginosa PD100 could act in the presence of SDS and Tween 80. This protease could be useful for degradation of protein in the presence of solvent, dehairing of cow skin and degradation of natural proteins. The immobilized protease showed 15-20% increases in temperature stability and the entrapped enzyme retained 83% of its initial activity after six cycles. With respect to properties of the enzyme and its capability for degradation of different protein sources, this protease finds potential application for waste treatment, used in detergents and leather industry.