Design, Fabrication and Characterization of Nanoliposomes Containing Snake Venom of Pseudocereaster percius

Background: Development of antivenom or antidote requires the repetition of immunization of large animals, such as horses and goats, which ultimately releases the IgG immunoglobulin produced in the serum specimen. As snake venom involves a variety of proteins and enzymes getting administered into the animal, this process can inflict significant harm to the animal, therefore choosing carriers that can deliver the least amount of venom could be a safer option for animal immunization Objective: In this research, nanoliposomes were used to encapsulate venom as a protected cargo for immunization. We used two distinct liposomal formulations to entrap the venom: 1,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-distearoyl-sn-glycero-3-phospho-(1′-rac-glycerol) associated with cholesterol in one formulation and dimethyldioctadecylamonium (Bromide salt) paired with cholesterol in the other. Method: Liposomal formulations prepared by solvent evaporation method and the venom was encapsulated in liposomes and evaluated for size and zeta potential. Meanwhile, encapsulation efficiency, venom release percentage, and phospholipase activity have all been analyzed. Results: The findings revealed that dimethyldioctadecylamonium (Bromide salt) combined with cholesterol had the highest encapsulation efficiency. In this formulation, the venom release rate had a steady-state profile. The lack of phospholipase activity in this formulation may be due to a bromide group in the liposomal structure that could be useful for immunization. Conclusion: Liposomal formulations, which do not have the active site of the snake venom enzymes, could be used for venom encapsulation.

Peroxiredoxin-6 is a Guardian of lung pathophysiologies

: The moonlighting protein, Prdx6 exhibits peroxidase activity, phospholipase activity and lysophosphatidylcholine acyl transferase (LPCAT) activity. Although it is ubiquitous in expression, its level is prominently high in the lung. Prdx6 has been known to be an important enzyme for the maintenance of normal lung physiologies including, anti-oxidant defense, lung surfactant homeostasis and cell signaling. Studies further unveiled that the altered activity (peroxidase or aiPLA2) of this enzyme is linked with various lung pathologies or diseases. In the present article, we attempted to address the various pathophysiologies or disease conditions (like lung ischemia, hyperoxia, lung cancer, emphysema and acute lung injury) wherein prdx6 is involved. The study implicates that Prdx6 could be used as a common drug target for multiple lung diseases. Important future insights have also been incorporated.

Synthesis of the Polyhydroquinoline Derivative 4-(2-Chloro-Phenyl)-2,7,7-Trimethyl5-oxo-1,4,5,6,7,8-Hexahydroquinoline-3-Carboxylic Acid Ethyl Ester: Antimicrobial and Enzyme Modulator

Multicomponent reactions are extremely relevant in green chemistry. They offer better conditions than traditional synthesis and are, therefore, used for many organic modifications. Recently, the synthesis of polyhydroquinolines has received much attention for its high pharmacological potential. In the present study, a polyhydroquinoline derivative was synthesized without the use of catalysts or solvents. The results of nuclear magnetic resonance and infrared spectroscopy demonstrated that the molecule was successfully synthesized. The molecule presents significant results of antimicrobial activity for the bacteria tested in the serial dilution method. It also increased the clotting time by 25.66 seconds for the highest dose and 12.66 seconds for the other doses tested. Prior incubation with the dose of 125 mg reduced the thrombolytic activity to 73%. The 125, 100, and 50 mg doses previously incubated with Bothrops moojeni venom inhibited approximately 30% of the phospholipase activity. The molecule was also able to reduce the cytotoxicity induced by proteases significantly. In conclusion, the molecule presents several biological properties, which highlights its pharmaceutical potential.

iPLA2-VIA is required for healthy aging of neurons, muscle, and the female germline in Drosophila melanogaster

Neurodegenerative disease (ND) is a growing health burden worldwide, but its causes and treatments remain elusive. Although most cases of ND are sporadic, rare familial cases have been attributed to single genes, which can be investigated in animal models. We have generated a new mutation in the calcium-independent phospholipase A2 (iPLA2) VIA gene CG6718, the Drosophila melanogaster ortholog of human PLA2G6/PARK14, mutations in which cause a suite of NDs collectively called PLA2G6-associated neurodegeneration (PLAN). Our mutants display age-related loss of climbing ability, a symptom of neurodegeneration in flies. Although phospholipase activity commonly is presumed to underlie iPLA2-VIA function, locomotor decline in our mutants is rescued by a transgene carrying a serine-to-alanine mutation in the catalytic residue, suggesting that important functional aspects are independent of phospholipase activity. Additionally, we find that iPLA2-VIA knockdown in either muscle or neurons phenocopies locomotor decline with age, demonstrating its necessity in both neuronal and non-neuronal tissues. Furthermore, RNA in situ hybridization shows high endogenous iPLA2-VIA mRNA expression in adult germ cells, and transgenic HA-tagged iPLA2-VIA colocalizes with mitochondria there. Mutant males are fertile with normal spermatogenesis, while fertility is reduced in mutant females. Mutant female germ cells display age-related mitochondrial aggregation, loss of mitochondrial potential, and elevated cell death. These results suggest that iPLA2-VIA is critical for mitochondrial integrity in the Drosophila female germline, which may provide a novel context to investigate its functions with parallels to PLAN.

Evaluation of proteinase and phospholipase activity of Candida albicans strains isolated from the mouth of patients with controlled and uncontrolled diabetes mellitus in Qazvin province: Short Communication

Pathophysiological changes in the mouth of patients with diabetes mellitus (DM) predispose to overgrowth of Candida albicans. Various virulence factors such as proteinase and phospholipase production are involved in the process of degradation of the host cell membrane. These enzymes are involved in binding Candida species, especially in the hyphae stage, to the target tissue. In this study, the proteinase and phospholipase activity of 51 C. albicans strains was evaluated. Overall, the rates of proteolytic and phospholipase activities in controlled and uncontrolled diabetic patients were 90.2% and 72.5%, respectively. No significant difference was observed between proteinase and phospholipase activities in both groups of patients (P>0.05), while there was a significant difference between the activity of hydrolytic enzymes with HbA1C and dentures in the uncontrolled diabetic group (P<0.05). According to the importance of hydrolytic enzymes activity of C. albicans species in diabetic patients, oral health of diabetic patients to control and prevent disease progression in these patients is essential.

iPLA2-VIA is required for healthy aging in neurons, muscle, and female germline in Drosophila melanogaster

Neurodegenerative disease (ND) is a growing health burden worldwide, but its causes and treatments remain elusive. Although most cases of ND are sporadic, rare familial cases have been attributed to single genes, which can be investigated in animal models. We have generated a new mutation in the calcium-independent phospholipase A2 (iPLA2) VIA gene CG6718, the Drosophila melanogaster ortholog of human PLA2G6/PARK14, mutations in which cause a suite of NDs collectively called PLA2G6-associated neurodegeneration (PLAN). Our mutants display age-related loss of climbing ability, a symptom of neurodegeneration in flies. Although phospholipase activity commonly is presumed to underlie iPLA2-VIA function, locomotor decline in our mutants is rescued by a transgene carrying a serine-to-alanine mutation in the catalytic residue, suggesting that important functional aspects are independent of phospholipase activity. Additionally, we find that iPLA2-VIA knockdown in either muscle or neurons phenocopies locomotor decline with age, demonstrating its necessity in both neuronal and non-neuronal tissues. Furthermore, RNA in situ hybridization shows high endogenous iPLA2-VIA mRNA expression in adult germ cells, and transgenic HA-tagged iPLA2-VIA colocalizes with mitochondria there. Mutant males are fertile with normal spermatogenesis, while fertility is reduced in mutant females. Mutant female germ cells display age-related mitochondrial aggregation, loss of mitochondrial potential, and elevated cell death. These results suggest that iPLA2-VIA is important for germline mitochondrial integrity in Drosophila, which may be relevant for understanding how PLAN develops.

Effect of Vernonia amygdalina on phospholipase activity from Naja Mossambica (Cobra)

Background: Phospholipases are one of the numerous enzymes found in the Naja mossambica venom. They play a major role in snakebite envenomation, and also responsible for the hydrolysis of a phospholipid, disrupting the membrane integrity. In this study, we evaluated the effect of Vernonia amygdalina on Phospholipase activity from Naja mossambica (Cobra) Results: Partially purified phospholipase had maximal velocity (Vmax) and Michaelis Menten constant (Km) of 7.6 × 10-5 mol/min and 1.7mg/ml, while the crude phospholipase had Vmax and Km of 9.4 × 10-5mol/min and 2.5mg/ml respectively. Inhibition study of aqueous extracts of Vernonia amygdalina leaf shows that the extract is a potent inhibitor of crude phospholipase in a dose-dependent pattern. The different doses of extract 15 %, 10 % and 5% produced percentage inhibition of 74.04 %, 78.6 % and 86.63% respectively. The kinetic binding constant (Ki) values of crude phospholipase for different concentrations of extracts 5%, 10% and 15% were 0.21mg/ml, 0.29mg/ml and 0.39mg/ml, while the partially purified phospholipase for different concentrations of extracts 5%, 10% and 15% were were 0.48mg/ml, 0.42mg/ml and 0.41mg/ml respectively. It can be deduced from the results that the extract inhibits the phospholipase activity in an uncompetitive manner. Conclusion: Aqueous extract of Vernonia amygdalina leaves may contain some bioactive agents that could serve as potent inhibitors of phospholipase from Naja mossambica venom.

Exercise-Induced Alterations in Phospholipid Hydrolysis, Airway Surfactant and Eicosanoids and their Role in Airway Hyperresponsiveness in Asthma

The mechanisms responsible for driving endogenous airway hyperresponsiveness (AHR) in the form of exercise-induced bronchoconstriction (EIB) are not fully understood. We examined alterations in airway phospholipid hydrolysis, surfactant degradation, and lipid mediator release in relation to AHR severity and changes induced by exercise challenge. Paired induced sputum (n=18) and bronchoalveolar lavage (BAL) fluid (n=11) were obtained before and after exercise challenge in asthmatic subjects. Samples were analyzed for phospholipid structure, surfactant function and levels of eicosanoid and secreted phospholipase A2 group 10 (sPLA2-X). A primary epithelial cell culture model was used to model effects of osmotic stress on sPLA2-X. Exercise challenge resulted in increased surfactant degradation, phospholipase activity, and eicosanoid production in sputum samples of all patients. Subjects with EIB had higher levels of surfactant degradation and phospholipase activity in BAL fluid. Higher basal sputum levels of cysteinyl leukotrienes (CysLTs) and prostaglandin D2 (PGD2) were associated with direct AHR and both the post-exercise and absolute change in CysLTs and PGD2 levels were associated with EIB severity. Surfactant function was either abnormal at baseline or became abnormal after exercise challenge. Baseline levels of sPLA2-X in sputum and the absolute change in amount of sPLA2-X with exercise were positively correlated with EIB severity. Osmotic stress ex vivo resulted in movement of water and release of sPLA2-X to the apical surface. In summary, exercise challenge promotes changes in phospholipid structure and eicosanoid release in asthma, providing two mechanisms that promote bronchoconstriction, particularly in individuals with EIB who have higher basal levels phospholipid turnover.