An up-to-date review of biomedical applications of serratiopeptidase and its biobetter derivatives as a multi-potential metalloprotease.

Serratiopeptidase is a bacterial metalloprotease used in a variety of medical applications. The multidimensional properties of serratiopeptidase make it noticeable as a miraculous enzyme. Anti-coagulant, anti-inflammatory and anti-biofilm activity of serratiopeptidase making it useful in reducing pain and swelling associated with various conditions including arthritis, diabetes, cancer, swelling, pain and also thrombolytic disorders. It breaks down fibrin, thins the fluids formed during inflammation and due to its anti-biofilm activity, can be used in the combination of antibiotics to reduce development of antibiotic resistance. However, some drawbacks like sensitivity to environmental conditions and low penetration into cells due to its large size have limited its usage as a potent pharmaceutical agent. To overcome such limitations, improved versions of the enzyme were introduced using protein engineering in our previous studies. Novel functional serratiopeptidases with shorter length and higher stability have seemingly created a hope for using this enzyme as a more effective therapeutic enzyme. This review explains the structural properties and functional aspects of serratiopeptidase, its main characteristics and properties, pre-clinical and clinical applications of the enzyme, improved qualities of the modified forms, different formulations of the enzyme, and the potential future developments.

A surgical case of anti-coagulant ileus mimicking small-bowel tumors: a case report.

BACKGROUND: Anti-coagulant ileus, characterized by intramural hematoma due to excessive anti-coagulant therapy, presents a diagnostic challenge. Although previously considered uncommon, recently, reporting cases of anti-coagulant ileus have become more frequent. Herein, we report a rare surgical case of anti-coagulant ileus mimicking small-bowel tumors. CASE PRESENTATION: A 79-year-old man was admitted to our hospital for fatigue. He had been administered warfarin for 5 months for atrial fibrillation. On admission, the patient exhibited mild epigastric tenderness. Laboratory test results revealed anemia (hemoglobin, 8.4 g/dL); unmeasurably prolonged prothrombin time (PT) with international normalized ratio (INR) > 8; and elevated soluble interleukin 2 receptor (sIL-2R) levels (849 IU/mL; normal range, 122-496 IU/mL). Abdominal plain computed tomography (CT) showed a circumferentially thickened intestinal wall at one site in the jejunum and two in the ileum. After hospitalization, bowel obstruction did not improve with conservative treatment. Suspecting small-bowel tumors such as lymphoma, the patient subsequently underwent open surgery on day 3 after admission. No obvious tumor mass was observed intra-operatively. However, only thickened and hemorrhagic segments were identified at the suspected sites. We performed partial jejunal and ileal resections of 12 and 27 cm, respectively. Histopathology confirmed submucosal congestion, edema, and hemorrhage in each area without tumor components, leading to the final diagnosis of intramural hematoma. The postoperative course was uneventful, and he was discharged on postoperative day 9. No recurrence occurred during the 5-year follow-up period. CONCLUSIONS: We encountered a surgical case of anti-coagulant ileus, which was difficult to differentiate from malignant lymphoma based on CT findings and high sIL-2R levels. The possibility of anti-coagulant ileus should always be considered in patients on long-term anticoagulation medication and bowel obstruction with high PT-INR values.

Efficacy of unfractionated heparin in patients with moderate sepsis-induced coagulopathy: An observational study.

INTRODUCTION: The 2021 Surviving Sepsis Campaign guidelines recommend low-molecular-weight heparin for the prevention of venous thromboembolism in sepsis. However, observational studies suggest that anticoagulants as a whole may benefit severely ill sepsis patients with coagulopathy, but the optimal targets of unfractionated heparin remain unclear. This study investigated which sepsis patients could most benefit from unfractionated heparin. MATERIALS AND METHODS: In this retrospective observational study, we identified adult sepsis patients requiring urgent hospitalization from 2006 to 2019 using a large-scale Japanese medical database. Patients were divided into two groups: those receiving unfractionated heparin within 72 h of admission and those who did not. We compared in-hospital mortality, major bleeding complications, and thromboembolic events between these groups using a multivariate logistic regression model adjusted for patient and treatment variables. Additionally, we assessed the association between heparin administration and in-hospital mortality across various subgroups. RESULTS: Among 30,342 sepsis patients, 2520 received early heparin administration, and 27,822 did not. Multivariate logistic regression revealed a significant association between heparin and reduced in-hospital mortality (adjusted OR: 0.735, 95 % CI: 0.596-0.903) but no significant association with major bleeding and thromboembolic risk (adjusted OR: 1.137, 1.243; 95 % CI: 0.926-1.391, 0.853-1.788, respectively). Subgroup analyses suggested significant survival benefits associated with heparin only in the sepsis patients with moderate coagulopathy and sepsis-induced coagulopathy scores of 3 or 4 (adjusted OR: 0.452, 0.625; 95 % CI: 0.265-0.751, 0.410-0.940, respectively). CONCLUSIONS: Early heparin administration upon admission is associated with lower in-hospital mortality, especially in moderate sepsis-induced coagulopathy, and no significant increase in complications.

Evaluating anthelmintic, anti-platelet, and anti-coagulant activities, and identifying the bioactive phytochemicals of Amaranthus blitum L.

BACKGROUND: Highlighting affordable alternative crops that are rich in bioactive phytoconstituents is essential for advancing nutrition and ensuring food security. Amaranthus blitum L. (AB) stands out as one such crop with a traditional history of being used to treat intestinal disorders, roundworm infections, and hemorrhage. This study aimed to evaluate the anthelmintic and hematologic activities across various extracts of AB and investigate the phytoconstituents responsible for these activities. METHODS: In vitro anthelmintic activity against Trichinella spiralis was evaluated in terms of larval viability reduction. The anti-platelet activities were assessed based on the inhibitory effect against induced platelet aggregation. Further, effects on the extrinsic pathway, the intrinsic pathway, and the ultimate common stage of blood coagulation, were monitored through measuring blood coagulation parameters: prothrombin time (PT), activated partial thromboplastin time (aPTT), and thrombin time (TT), respectively. The structures of isolated compounds were elucidated by spectroscopic analysis. RESULTS: Interestingly, a previously undescribed compound (19), N-(cis-p-coumaroyl)-ʟ-tryptophan, was isolated and identified along with 21 known compounds. Significant in vitro larvicidal activities were demonstrated by the investigated AB extracts at 1 mg/mL. Among tested compounds, compound 18 (rutin) displayed the highest larvicidal activity. Moreover, compounds 19 and 20 (N-(trans-p-coumaroyl)-ʟ-tryptophan) induced complete larval death within 48 h. The crude extract exhibited the minimal platelet aggregation of 43.42 ± 11.69%, compared with 76.22 ± 14.34% in the control plasma. Additionally, the crude extract and two compounds 19 and 20 significantly inhibited the extrinsic coagulation pathway. CONCLUSIONS: These findings extend awareness about the nutritional value of AB as a food, with thrombosis-preventing capabilities and introducing a promising source for new anthelmintic and anticoagulant agents.

Phytochemical characterization and phospholipase A2 inhibitory effect of Vitex negundo L. root extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Snake bites are a critical health issue in many parts of the world particularly in Asian countries lacking efficient health facilities in rural areas. Cobra is the most common snake type in Asia and is responsible for a large number of mortalities particularly in rural areas. Plants are usually considered the most effective and easy-to-approach treatment for snake bites in rural areas of various countries. Vitex negundo L. is an important medicinal plant traditionally used to treat snake bite envenomation in many countries of Asia. AIM OF THE STUDY: From literature survey of plants traditionally used in the treatment of snake bites in Asian countries including India, Pakistan and Sri Lanka, roots of V. negundo were selected for the present study. Anti-snake venom potential of its roots was assessed through various in vitro assays targeting the phospholipase A2 enzyme. MATERIALS AND METHODS: V. negundo roots were sequentially extracted in different organic solvents to get fractions and in methanol to get total extract. The extracts were evaluated for phospholipase A2 (PLA2) inhibitory potential through inhibition of venom-induced hemolysis, ADP-induced platelet aggregation, PLA2-induced fatty acid hydrolysis and anticoagulant effect of cobra venom. Antioxidant power was determined using DPPH and superoxide radical scavenging assays. GC-MS and HPLC analysis was performed for the total methanol extract. RESULTS: Strong PLA2 inhibitory effect was observed for all the extracts. The ethyl acetate, acetone and methanol fractions significantly inhibited toxic effects of cobra venom under in vitro conditions. Radical scavenging potential of these fractions was also significantly high as compared to non-polar fractions in both DPPH and superoxide scavenging assays. Phytochemical analysis indicated high phenolic and flavonoid contents in these fractions. GC-MS and HPLC analysis of total methanol extract confirmed the presence of bis(2-ethylhexyl) phthalate, phenol, o-Guaiacol, palmitic acid-methyl ester, methyl stearate, quercetin and kaempferol in the plant. CONCLUSION: The study concluded that the roots of V. negundo, particularly their polar extracts, have strong PLA2 inhibitory effect against cobra venom confirming their traditional use to manage snake bites. The roots of this plant can be further studied for isolation of plant-based antisera.

Reversing the Tide: A Case of a Mechanical Aortic Valve Recipient Lost to Follow-up, Education on Rivaroxaban Contraindications, and the Vital Role of Acenocoumarol in Preventing Valve Thrombosis.

Aortic stenosis is the most common heart valve disease, especially among the elderly. Symptomatic aortic valve stenosis is linked to a poor prognosis and a high mortality rate if left untreated. The only effective treatment for severe symptomatic aortic stenosis is aortic valve replacement using either a mechanical or a biological prosthesis. Mechanical valve prostheses, while highly durable, are thrombogenic, necessitating lifelong anticoagulation with oral anti-vitamin K agents, such as acenocoumarol. Conversely, bioprosthetic valves, though less durable, carry a minimal thrombogenic risk and do not require anticoagulation. Currently, there is no proven role for direct-acting oral anticoagulants (DOACs) in patients with mechanical heart valves due to insufficient clinical trial data regarding their safety in this patient population. Herein, we present the case of a 59-year-old female known to have aortic stenosis, who underwent surgical treatment with mechanical aortic valve replacement eight years ago. Post-surgery, acenocoumarol was initiated. However, 18 months prior to presenting at our institution, the patient started taking rivaroxaban (a DOAC) instead of acenocoumarol due to the unavailability of acenocoumarol during the ongoing economic crisis in Lebanon, without consulting her cardiologist. Although she was followed up by her general practitioner and reported having a mechanical valve, her son contradicted this, claiming she had a biological valve. After thorough investigations, including chest X-ray, echocardiography, and fluoroscopy, it was confirmed that the patient indeed had a normally functioning mechanical aortic valve. Immediate corrective measures were taken, starting with IV unfractionated heparin and acenocoumarol, targeting an International Normalized Ratio (INR) between 2.5 and 3, while educating the patient about her condition and the importance of adhering to acenocoumarol therapy.

In Vitro Screening of Antimicrobial and Anti-Coagulant Activities, ADME Profiling, and Molecular Docking Study of Citrus limon L. and Citrus paradisi L. Cold-Pressed Volatile Oils.

Citrus, which belongs to the Rutaceae family, is a very widespread genus in the Mediterranean Basin. In Tunisia, various parts of these spontaneous or cultivated plants are used in common dishes or in traditional medicine. The purpose of this work was to investigate C. limon and C. paradisi essential oil (EO). The samples were studied for their chemical composition using SPME/MS, as well as their antibacterial and antifungal activities. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) methods were used to evaluate the anticoagulant potentialities. The obtained results show that both essential oils are rich in monoterpenes hydrocarbons, whereby limonene is the main compound in C. paradisi EO (86.8%) and C. limon EO (60.6%). Moreover, C. paradisi EO contains ß-pinene (13.3%), sabinene (2.2%) and α-pinene (2.1%). The antibacterial assay of the essential oils showed important bactericidal and fungicidal effects against all strains tested. In fact, the MICs values of C. limon EO ranged from 0.625 to 2.5 mg/mL against all Gram-positive and Gram-negative bacteria, and from 6.25 to 12.5 mg/mL for Candida spp. strains, while C. paradisi EO was more active against all bacteria with low MICs values ranging from 0.192 to 0.786 mg/mL, and about 1.5 mg/mL against Candida species. Both tested Citrus EOs exhibited interesting anticoagulant activities as compared to heparin. The molecular docking approach was used to study the binding affinity and molecular interactions of all identified compounds with active sites of cytidine deaminase from Klebsiella pneumoniae (PDB: 6K63) and the C (30) carotenoid dehydrosqualene synthase from Staphylococcus aureus (PDB: 2ZCQ). The obtained results show that limonene had the highest binding score of -4.6 kcal.mol-1 with 6K63 enzyme, and -6.7 kcal.mol-1 with 2ZCQ receptor. The ADME profiling of the major constituents confirmed their important pharmacokinetic and drug-like properties. Hence, the obtained results highlight the potential use of both C. limon and C. paradisi essential oils as sources of bioactive compounds with antibacterial, antifungal, and anti-coagulant activities.

Dental management of patients on anti-thrombotic agents

The number of geriatric patients seeking dental service is ever-rising because of increased life expectancy, also with problem of increased chronic medical conditions. One of them are patients on anti-thrombotic medication. Bleeding complication after minor oral surgery by anti-thrombotic agents is of concerns to dentists on dental management of these patients. Risk and benefit of the anti-thrombotic agents must be weighed before initiating dental procedures, which should be established as a treatment guideline. Purpose of the paper is to optimize the management of the dental patients on anti-thrombotic medication via standardization of treatment protocol of such a patient.

Clinical characteristics and treatments of anti-coagulant rodenticide poisoning in children lacking of poisoning history / 中国小儿急救医学

Objective To summarize the clinical characteristics,diagnosis and treatments of 50 chil-dren with no history of anti-coagulation rodenticide poisoning.Methods Fifty children lacking of poisoning history were admitted to our hospital from Feb 2011 to Dec 2015,who were due to hemorrhage and finally di-agnosed as rodenticide poisoning.The clinical manifestation,characteristics of blood coagulation dysfunction, toxicology screening results and treatments of 50 cases were retrospectively analyzed.Results All 50 cases were from rural area,and denied the history of taking rodenticide by mistake.The time from the onset to ad-mission to the hospital was from 1 d to 120 d,with an average of 11.66 d.Major clinical manifestations in-cluded:34 cases(68%)of skin ecchymosis,26 cases(52%)of epistaxis,10 cases(20%)of bleeding gums and 4 cases(8%)of hematoma at the injection site,with PT 20 to over 200 s and APTT 31 to over 200 s (maximal detection value).The children were diagnosed as rodenticide poisoning when one of brodifacoum, bromadiolone,and diphacinone-sodium or combination of brodifacoum and bromadiolone were detected from their blood specimens,with an concentration of 5 to 3 270 μg/L.The diagnosed children were intravenous dripped with specific antagonist vitamin K1 and the critically ill children were added fresh frozen plasma.All children recovered from hemorrhage within 24 hours after treatment.The level of PT,APTT was close to nor-mal after 3 to 7 days of treatment,and the using of vitamin K1 was accumulated about 30 to 70 mg.The chil-dren were given maintenance treatment of vitamin K1 for three months after discharge until the condition was stable.Conclusion Commonly used rodenticide has the characteristics of fat soluble, easy to accumulate, long incubation period,and long treatment period and so on.Children with no bleeding performance have the clinical tendency of acute bleeding,prolonged PT and APTT should be considered the possibility of rodenti-cide poisoning.Blood or urine samples should be screened for toxicology as soon as possible,and meanwhile children should be given antagonist treatment with special antidote vitamin K1 until the coagulation function is stable in normal range to avoid recurrence of the illness.

In vitro study of inhibitory effect of Ulinastatin against serine protease activity / 中国生化药物杂志

Objective To study inhibitory effect of serine protease activity by Ulinastatin in vitro .Methods Different chromogenic peptides were designed and synthesized.Highly sensitive fluorescence detection was performed to optimize the concentration of each serine proteases and their chromogenic substrates.Multi-point method was used for the calculation of half maximal inhibitory concentration of Ulinastatin .ResuIts Ulinastain could inhibit Polymorphonuclear leukocyte elastase ( PMNE ) and plasmin with IC50 lower than 100 U/mL.For factor Xa, and Kallikrein, the IC50 of Ulinastatin was higher than 1000U/mL.No thrombin IC50 could be calculated at the present experiments.ConcIusion Similar to Ulinastatin injection from Japan, domestic Ulinastatin shows the strongest inhibitory effects on PMNE among those serine proteases.As important references, this study gives reliable data for dose range of domestic Ulinastatin in anti-inflammation, coagulation/anti-coagulation and anti-shock therapy.

Laser Thermal Angioplasty by the Contact Method for Arteriosclerosis Obliterans of the Lower Limbs / 日本心臓血管外科学会雑誌

Laser thermal angioplasty with a metal hot tip probe was performed on 34 cases of arteriosclerosis obliterans of the lower limbs, instead of conventional balloon angioplasty. Initial angiographic and clinical success was achieved in 48 of 53 (90.5%) arterial lesions. The initial success rates were 91.6%, 87.8% and 100% for lesions of the iliac, femoral and popliteal arteries respectively. The initial success rate was 66.6% in occlusive lesions, and was 100% in stenotic lesions. Five of seven (71.4%) occlusions less than 10cm length, and five of eight (62.5%) occlusions longer than 10cm were successfuly treated. Higher laser energy was necessary for ablation in occlusive lesions than in stenotic lesions. The ankle pressure index improved postoperatively in both occlusive and stenotic lesions. Satisfactory long term patency was achieved in 27 of 35 successfully treated lesions (77.1%). These results suggest that laser thermal angioplasty is effective for the treatment of arteriosclerotic disease in the lower limbs.

Anticardiolipin Antibody (ACA) and Lupus Anticoagulant (LA): Association with Vascular Access Occlusion in Hemodialysis (HD) Patients / 대한내과학회지

OBJECTIVES: Anticardiolipin antibody (ACA) and lupus anticoagulant (LA) are acquired antiphospholipid antibodies (APAs), which are regarded as important risk factors far vascular thrombosis and recurrent fetal loss. Although the clinical relevance of APAs in dialysis patients is uncertain, recent studies have suggested that APAs are involved in bioincompatibility and thrombogenic complications in hemadialysis (HD) patients. METHOD: We performed a cross sectional study of ACA and LA in 50 stable HD patients and their 68 vascular accesses (52 native arteriovenous fistulae and 16 synthetic arterovenous grafts), with the analysis of factors associated with the presence of APAs and the retrospective evaluation of vascular access occlusion (VAO). LA was assessed by platelet neutralization method whereas IgG-ACA was measured by a solid phase ELISA. Values higher than 23GPLU/ml (IgG phospholipid units) were considered to be positive for IgG-ACA and positive values for LA was more than 8 seconds in prolongation of the clotting time with human platelet lysate. Vascular access survival was assessed by Kaplan- Meier method, RESULTS: The mean age of the subject (M:F 21:29) was 46 years and the mean duration of hemodialysis was 49 months. The frequency of VAO in entire subjects was 0.45+/-0.98 episodes/patient year. The median value of IgG-ACA was 16.0 GPLU/ml with a distribution from 2.7 to 46.1GPLU/ ml. The median titer of I.A was 4.5 (3.1-45.6) seconds. Fourteen patients (28%) were found to have at least one episode of VAO. In spite of comparable clinical and biochemical data according to the presence of VAO, the titers of IgG-ACA (13.6+/-7.7 vs, 20.3+/-8.7GPLIJ/ml, P<0.05) and LA (4.5+/-2.9 vs. 11.7 +/-12.6sec, P<0.05) were significantly higher in VAO group. Six out of 50 patients(12%) had an increased titer of IgG-ACA and LA was found in 11 patients(22%). No patients were positive for ACA and LA simultaneously. There was no significant difference in sex, etiology of ESRD, diabetic status, the dosage of heparin during HD or the amount of erythropoietin administered according to the presence of APAs. We could not find any significant correlation between the titer of APAs and age, duration of dialysis, blood pressure, platelet count and biochemical parameters. In the patients with positive ACA, the frequency of VAO was 1.05+/-0.12 episodes/patient year, which was significantly higher than patients without ACA (0.33+/-0.17 episodes/ patient year, P<0.05). In the patients with the presence of LA(1.06+/-0.43 vs. 0.12+/-0.06 episodes/ patients year, P<0.01). The median vascular access survival time in IgG-ACA positive patients (32.7 months) was significantly decreased compared to 66.8 months in IgG-ACA negative group. CONCLUSION: Our data suggest that the presence of APAs (ACA and/or LA) affects the event-free vascular access survival in HD patients. Therefore the evaluation of APAs status have to be included in the diagnostic strategies for the patients with recurrent VAO. Further studies are necessary to explore the pharmacologic intervention method to decrease APAs and prevent VAO in HD patients.