Tamarind Trypsin Inhibitor in Chitosan–Whey Protein Nanoparticles Reduces Fasting Blood Glucose Levels without Compromising Insulinemia: A Preclinical Study

In vivo studies show the benefits of the trypsin inhibitor isolated from tamarind (Tamarindusindica L.) (TTI) seeds in satiety and obesity. In the present study, TTI nanoencapsulation (ECW) was performed to potentialize the effect of TTI and allow a controlled release in the stomach. The impact on glycemia, insulin, and lipid profile was evaluated in Wistar rats overfed with a high glycemic index diet (HGLI). Characterization of the nanoparticles and in vitro stability in simulated gastrointestinal conditions, monitored by antitrypsin activity and HPLC, was performed. ECW and empty nanoparticles (CW) were administered by gavage, using 12.5 and 10.0 mg/kg, respectively. Both nanoformulations presented a spherical shape and smooth surface, with an average diameter of 117.4 nm (24.1) for ECW and 123.9 nm (11.3) for CW. ECW maintained the antitrypsin activity (95.5%) in the gastric phase, while TTI was completely hydrolyzed. In Wistar rats, the nanoformulations significantly reduced glycemia and HOMA IR, and ECW increased HDL-c compared to CW (p < 0.05).Pancreas histopathology of animals treated with ECW suggested an onset of tissue repair. Thenanoencapsulation provided TTI protection, gradual release in the desired condition, and improvement of biochemical parameters related to carbohydrate metabolism disorders,without compromising insulinemia.

Changes of nonspecific proteinases and their inhibitors in different clinical types of myocardial infarction

Purpose: to elucidate the relationships between serum levels of nonspecific proteinases and their inhibitors in patients with complications of myocardial infarction (MI). Material and methods. This prospective short-term study included 82 patients with uncomplicated non-Q wave MI (n=27), Killip I-II Q-MI complicated by acute left ventricular failure (ALVF) (n=30), Killip III-IV Q-MI, and Killip III-IV ALVF (n=17) as well as non-survivors due to development of cardiogenic shock (n=8) and healthy controls (n=12). Serum levels of elastase-like (ELA) and trypsine-like (TLA) activities and of proteinase inhibitors (antitrypsin activity and acid-stable inhibitors) was evaluated by enzymatic method within 24 hours, 3 and 14 days after the onset of symptoms. Risk stratification was done at admission according to the GUSTO Score. Results. The levels of all nonspecific proteinases were elevated in MI patients in comparison to controls. Mean baseline ELA (0.29 vs 0.37 nMol/mL min; р<0.05) and TLA (0.33 vs 0.40 mcMol/mL min; р<0.05) were higher in patients with MI complicated by ALVF than in those with uncomplicated MI. Non-survivors showed significantly higher ELA and TLA and lower levels of proteinase inhibitors than patients with uncomplicated MI. Discussion. Proteolytic enzyme activities varied parallel to the changes of proteinase inhibitor levels. Proteinase activities were much more elevated in non-survirvors than in patients with uncomplicated MI. Decreased proteinase inhibitor levels can be considered as a marker of imbalance of proteinase-inhibitor system. Conclusions. Increase of nonspecific proteinase serum levels within 24 hours after the onset of MI is associated with the development of ALVF and unfavorable prognosis; it suggests the involvement of the proteinase-inhibitor system in pathogenesis of MI.

Pathogenetic significance of proinflammatory cytokines, nonspecific proteinases and their inhibitors for the risk of complications of acute myocardial infarction

Цель исследования состояла в изучении взаимосвязи между тяжестью течения заболевания и реакцией систем провоспалительных цитокинов, неспецифических протеиназ и их ингибиторов у больных острым инфарктом миокарда (ИМ) при краткосрочном наблюдении. Методы. Обследовано 82 пациента, из них 27 пациентов с неосложненным ИМ без зубца Q, 30 пациентов с Q-ИМ, осложненным острой левожелудочковой недостаточностью (ОЛЖН) I-II класса по Киллипу, 17 пациентов с Q-ИМ и ОЛЖН III-IV класса, 8 пациентов с летальным исходом вследствие развития рефрактерного кардиогенного шока (КШ). Контрольную группу составили 12 практически здоровых лиц. На 1-е, 3-и и 14-е сутки от начала развития ИМ определяли содержание в крови интерлейкина 1b (ИЛ-1b), интерлейкина 6 (ИЛ-6), фактора некроза опухоли a (ФНО-a), показатели неспецифических протеиназ и их ингибиторов. Результаты. В первые сутки ИМ уровни всех цитокинов значительно превышали показатели контрольной группы. Концентрация ИЛ-6 в крови у пациентов с ИМ, осложненным ОЛЖН, была достоверно выше, чем в группе ИМ без зубца Q (27,5 ± 1,8 пг/мл и 16,0 ± 1,3 пг/мл соответственно; р<0,001). Аналогичная ситуация была выявлена и для ФНО-a (24,7 ± 2,9 и 19,6 ± 1,4 пг/мл соответственно; р<0.001), в то время как уровни ИЛ-1b существенно не различались между группами. Активация протеолитических ферментов развивалась параллельно с увеличением уровня их ингибиторов при неосложненных формах ИМ, а у пациентов, умерших вследствие КШ, активация протеиназ отмечалась на фоне уменьшения уровня их ингибиторов. В группе с КШ также отмечены наиболее высокие показатели ИЛ-6 и ФНО-a. Заключение. Увеличение уровней ИЛ-6 и ФНО-a в крови в первые сутки ИМ и нарастание активности неспецифических протеиназ в динамике его развития сопровождаются развитием ОЛЖН с высокой вероятностью летального исхода, что может быть использовано в качестве дополнительного предиктора для оценки степени риска при ИМ. Purpose of this investigation was to evaluate the interrelationships of proinflammatory cytokines and nonspecific proteinases serum levels with the development of myocardial infarction (MI) complications. Methods. Eighty two patients with MI were studied. Prospective short-term study included patients with uncomplicated non-Q wave MI (n = 27), patients with Q-MI complicated by acute left ventricular insufficiency I-II Killip (n = 30), patients with Q-MI complicated by acute left ventricular insufficiency III-IV Killip (n = 17), non-survivors due to development of cardiogenic shock (n = 8) and age and gender matched healthy controls (n = 12). Serum levels of IL-1b, IL-6 and TNF-a had been evaluated by means of ELISA method, also was determine the elastase-like (ELA) and trypsine-like (TLA) activities and level of proteinase inhibitors (antitrypsin activity and acid-stable inhibitors). Blood samples had been drawn on admission to the hospital within 24 hours from the onset of symptoms, at 3d and 14th days of MI. Results. All cytokines levels were significantly elevated in MI patients in comparison to controls. Mean concentrations of IL-6 at baseline were higher among patients with MI complicated by acute left ventricular insufficiency than in group with uncomplicated MI (27.6 vs 16.0 pg/mL; р<0.001). The same was revealed in concentration of TNF- a (24.7 vs 19.6 pg/mL; р<0.01), while mean concentration of IL-1b did not differ significantly between these two groups. Non-survivors also showed significantly higher levels of IL-6, TNF-a, ELA and TLA and lower level of proteinase inhibitors than patients with uncomplicated MI. Activation of proteolytic enzyme activity developed together with changes level of proteinase inhibitors. In some cases level of elastase activity increases in 2-3 times. Conclusion: Increased levels of IL-6 and TNF-a, as well as nonspecific proteinases serum levels within 24 hours from the onset of MI are associated with the development of ALVF and poor prognosis and indicate participation of proteinase-inhibitory system in pathogenesis of MI. The data support the role of excessive cytokine-mediated inflammation in worsening of MI course and outcome.

Altered elastase—alpha1-antitrypsin balance in the blood of patients with chronic venous disease

Objectives Although leukocyte elastase is suspected to be involved in the damage of vein wall during chronic venous disease, the equilibrium between this protease and its inhibitor, alpha1-antitrypsin, has not yet been evaluated. The aim of the present study was to determine the relationship between leukocyte elastase and alpha1-antitrypsin, in the blood of patients with chronic venous disease. Patients and methods The concentration and the activity of leukocyte elastase along with the activity of alpha1-antitrypsin were evaluated in the blood of 55 chronic venous disease patients. The results were compared with those obtained in 33 healthy age and sex-matched volunteers. Results A significant decrease in the leukocyte elastase activity that correlated with an increased alpha1-antitrypsin activity was observed in the serum of patients with mild clinical symptoms of chronic venous disease. Conclusions The results of the study did not confirm a hypothesis about an important role of proteolytic activity of leukocyte elastase in the vein wall injury mechanism. They show that the leukocyte elastase–alpha1-antitrypsin balance is rather shifted toward antiprotease activity, especially in an early stage of chronic venous disease.