Enzymes In Tissues Research Articles

Isolation of Microglia and Analysis of Protein Expression by Flow Cytometry: Avoiding the Pitfall of Microglia Background Autofluorescence.

Microglia are a unique type of tissue-resident innate immune cell found within the brain, spinal cord, and retina. In the healthy nervous system, their main functions are to defend the tissue against infectious microbes, support neuronal networks through synapse remodeling, and clear extracellular debris and dying cells through phagocytosis. Many existing microglia isolation protocols require the use of enzymatic tissue digestion or magnetic bead-based isolation steps, which increase both the time and cost of these procedures and introduce variability to the experiment. Here, we report a protocol to generate single-cell suspensions from freshly harvested murine brains or spinal cords, which efficiently dissociates tissue and removes myelin debris through simple mechanical dissociation and density centrifugation and can be applied to rat and non-human primate tissues. We further describe the importance of including empty channels in downstream flow cytometry analyses of microglia single-cell suspensions to accurately assess the expression of protein targets in this highly autofluorescent cell type. This methodology ensures that observed fluorescence signals are not incorrectly attributed to the protein target of interest by appropriately taking into account the unique autofluorescence of this cell type, a phenomenon already present in young animals and that increases with aging to levels that are comparable to those observed with antibodies against highly abundant antigens.

Оптимизация протокола получения культуры дермальных фибробластов крыс

Introduction. Obtaining primary cultures of fibroblasts is necessary for conducting experimental studies to investigate the basic cell response mechanisms to various stimuli. Despite the widespread use of fibroblast cultures, methods for obtaining them from the skin are not standardized. The aim of the studywas to find the optimal conditions to obtain a fibroblast culture from rat skin using enzymatic tissue disaggregation. Materials and methods. The fibroblast culture was obtained from the dermis of 18 male Wistar rats (N=12, 5–6-week-old, and 60–65 g body weight; N=6, 8–10-week-old rats, and 160–180 g body weight). A series of experiments was carried out to obtain a fibroblast culture with selecting the collagenase type and defining its concentration and exposure time. Results.An optimized protocol to obtain rat dermis fibroblasts is presented. Moreover, the problems of obtaining a culture and practical aspects of its use are discussed. Conclusion. When obtaining a primary culture of rat skin fibroblasts we should consider the following fac-tors: the type of the enzyme, its concentration, and exposure time; the age of the animals; the area of skin graft collection. In young rats, the optimal result was achieved when the cells were isolated from the axillary zone using collagenase type II at a 1 mg/ml concentration during a 90-minute exposure. In adult animals, the enzymatic effect of collagenase type II on the skin graft obtained from the back was optimal at a 5 mg/ml concentration during a 120-minute exposure. Keywords: fibroblast culture, dermis, preparation protocols, rats

Mathematical Study of Medicine Propagation in Biological Tissue and Some of Its Applications

The paper deals with the problem of the distribution of the medicine (enzyme) in the damaged biological tissue where the reaction enzyme—substrat takes place. The biological problem is reduced to a singular degenerate initial-boundary value problem for two coupled ordinary differential equations. Analytical solution of the singular degenerated IBV-problem was obtained by power series. The solution demonstrates the real situation and found suitable to depict the degeneration of singular system, caused by low concentration of the enzyme.

A highly butyrylcholinesterase selective red-emissive mitochondria-targeted fluorescent indicator imaging in liver tissue of mice

Abstract A long-emitting (λex = 580 nm, λem = 628 nm) mitochondrial targeting butyrylcholinesterase (BChE) selective fluorescent chemosensor (BChE-NBD) has been synthesized. A prominent fluorescence enhancement only for BChE was found in aqueous solution. BChE-NBD has low biological toxicity and has been successfully applied to the detection and imaging of BChE in mitochondria in a variety of cells and not interfered by other analytes (especially acetylcholinesterase). More importantly, imaging study of the distribution of the BChE in mice was performed first time, which confirmed the widespread existence of the enzyme in liver tissue. This making the probe BChE-NBD hopeful to be an effective tool for investigates liver diseases in basic medicine and clinical applications.

Change in Proteolytic Profile in Heifers After Oligofructose Overload.

Laminitis in cattle is an important underlying cause of lameness, which leads to a significant reduction in economic and animal welfare. Nevertheless, the disordered pathological processes of laminitis remain unclear. Several proteinases are probably involved in the disorder of basement membrane (BM) metabolism in laminitis, for instance, matrix metalloproteinases (MMPs), neutrophil elastase (NE), and myeloperoxidase (MPO). This study aimed to investigate the change in proteolytic profile in circulating and lamellar tissues using an oligofructose (OF) overload-induced laminitis model in heifers. Twelve clinically healthy and nonlame Chinese Holstein heifers were recruited and randomly divided into two groups: OF-induced and control (CON). The OF-induced heifers group (n = 6) was administered 17 g/kg of body weight (BW) of OF dissolved in 2 L/100 kg of BW of tap water via the oral-rumen tube. The CON group (n = 6) was given an equal volume of tap water. The plasma samples were collected 0, 6, 12, 18, 24, 36, 48, 60, and 72 h after administration, and the lamellar samples were collected 72 h after euthanasia. The plasma samples were analyzed by zymography and reverse zymography. Histological examination, zymography, reverse zymography, and Western blot of lamellar samples were conducted. In the plasma of the OF-induced group, the pro-MMP9 activity increased from 36 h (P < 0.001) to 60 h (P < 0.05). Moreover, the plasma tissue inhibitors of metalloproteinase 1 (TIMP1) activity decreased after 18 h (P < 0.05), while the ratio of pro-MMP9 to TIMP1 and TIMP2 increased after 18 h (P < 0.001) and 48 h (P < 0.05), respectively. The act-MMP2, pro-MMP9, and act-MMP9 activities increased in the lamellar tissue of the OF-induced group compared with the CON group (P < 0.05). In addition, the expression of lamellar NE protein was higher in the OF-induced group (P < 0.01), while no change was found in lamellar MPO protein compared with the CON group. In conclusion, increased pro-MMP9 combined with decreased TIMP1 activity in the circulation might have caused the activation of blood neutrophils, while the activation of proteolytic enzymes in lamellar tissue probably led to the dysfunction of BM in the OF-induced group.

Daily oxygen supplementation to the incubator at different stages of embryonic development alters the activity of antioxidant enzymes in the lung tissue of broiler chicks at a high altitude

ABSTRACT 1. Pulmonary hypertension, associated with mitochondrial function in the lung tissue of broilers, can occur at hypoxic high altitudes. The present research examined the impacts of O2 supplementation at different embryonic stages on broiler organ development and antioxidant enzyme activities. 2. In total, 360 eggs from Ross 308 broiler breeders at sea level were divided into a control group (O2 non-supplementation group) and three experimental groups daily 1 h 23.5% O2 supplementation from days 0 to 11 (O0-11), from days 12 to 21 (O12-21), and from days 18 to 21 (O18-21) of embryonic age. 3. The lung, heart, right ventricular (RV), and left ventricular (LV) + septum of newly hatched and seven day old chicks from every group were dissected and weighed. Antioxidant enzyme activities were examined in their lung tissue. 4. The lung weight did not change in any group. At hatching, the heart weight (g and %) was higher in the O12-21 and O18-21 groups than in the O0-11 and control groups, but the relative heart weight was the highest in the O18-21 group in comparison with the O12-21 group. 5. Superoxide dismutase (SOD) activity increased in all experimental groups at both ages, but glutathione peroxidase (GPx) activity increased only in seven day old chicks. Catalase (CAT) exhibited high activity in the O12-21 and O18-21 groups at hatching. In the seven day old chicks, while the CAT activity did not change in the O18-21 group, it decreased in the O0-11 group and increased in the O12-21 group. 6. Glutathione reductase (GR) activity did not change in the O18-21 group, but GR exhibited low activity in the O0-11 group and high activity in the O18-21 group in newly hatched chicks. The GR activity only decreased in the O18-21 group of seven day old chicks. 7. The results indicated that oxygen supplementation to the incubator caused alterations in the antioxidant enzyme activities in the lungs of broiler chicks, and this may have been in response to oxidative stress.

Pluronic® F127-mediated control of insulin release rates from NPH microcrystals and blood glucose depression in STZ-induced diabetic rats.

Introduction: Neutral protamine Hagedorn (NPH) insulin is an intermediate-acting basal insulin with a long history of clinical use, consisting native human insulin. Its rather undesirable action profile, characterized by a peak release within a few hours, followed by insufficient insulin delivery upon a single subcutaneous (s.c.) dose, is well-documented. This may have been caused by the inherent microcrystal structure involving the basic peptide protamine, as well as the presence of tissue enzyme activities that readily act on protamine at the injection site. This issue may be circumvented by utilizing thermosensitive, erodible Pluronic F127 (PF127) to modulate the kinetics of insulin release from NPH over a period of 24 hours in which the hydrogel is completely eroded. Methods: Previously, we have shown that insulin release rates in vitro from NPH/PF127 formulations (0-25% PF127) markedly reduced the initial insulin release, especially in the presence of enzyme activity that selectively degraded protamine at 1-5 U/mL. Insulin release over the course of 20 hours was better modulated in the presence of increasing PF127 content. In this study, the insulin formulations (0, 20, and 25% PF127) were administered s.c. (4 U/kg) to streptozotocin (STZ)-induced diabetic rats and blood glucose levels were monitored over 24 hours. Results: In vivo blood glucose depression profiles in STZ-induced diabetic rats exhibited a similar pattern of control to in vitro data at the single s.c. dose of 4 U/kg, apparently extending the duration of action of NPH over a 24-hour period in the presence of PF127. Conclusion: Our findings suggest that the undesirable kinetics of insulin release from NPH is significantly influenced by tissue enzyme activity and that the presence of PF127 provided a timely modulation of insulin release from NPH microcrystals in the STZ-induced diabetic rat model.

Correction: Effects of PKB/Akt inhibitors on insulin-stimulated lipogenesis and phosphorylation state of lipogenic enzymes in white adipose tissue.

Correction: Effects of PKB/Akt inhibitors on insulin-stimulated lipogenesis and phosphorylation state of lipogenic enzymes in white adipose tissue.

MicroRNA-30e up-regulation alleviates myocardial ischemia-reperfusion injury and promotes ventricular remodeling via SOX9 repression

AimAt present, studies have focused on microRNAs (miRNAs) in myocardial ischemia-reperfusion injury (MI/RI). But the specific role of miR-30e hasn’t been fully explored. Thus, this study is to uncover the mechanism of miR-30e in MI/RI. MethodsMI/RI models of rats and hypoxia/reoxygenation injury (H/R) models of H9C2 cardiomyocytes were established. Rats were injected with miR-30e and SRY-related high mobility group-box gene 9 (SOX9)-related oligonucleotides or vectors to explore their roles in MI/RI. H9C2 cardiomyocytes were transfected with restored miR-30e and depleted SOX9 to decipher their function in H/R injury. miR-30e and SOX9 expression in myocardial tissues and cardiomyocytes were detected. Online website prediction and luciferase activity assay were applied to validate the targeting relationship between miR-30e and SOX9. ResultsDecreased miR-30e and increased SOX9 were found in myocardial tissues of MI/RI rats and H/R-treated cardiomyocytes. miR-30e targeted SOX9. miR-30e up-regulation or SOX9 down-regulation reduced cardiac function damage and suppressed oxidative stress, inflammation, cardiomyocyte apoptosis and myocardial enzymes in myocardial tissues of MI/RI rats. Restoring miR-30e or silencing SOX9 energized cell viability and inhibited apoptosis of H/R-treated cardiomyocytes. Down-regulating SOX9 reversed the effects of miR-30e down-regulation on myocardial injury, ventricular remodeling, cardiomyocyte damage and apoptosis in MI/RI. ConclusionIt is concluded that miR-30e elevation alleviated cardiac function damage and promoted ventricular remodeling via SOX9 repression.

Acute Toxicity of Tuber Peels of Tacca leontopetaliodes on the Biochemistry and Histology of Gills and Liver of Clarias gariepinus Juveniles

Background: Increased rate of water pollution has detrimental effects on the health of fish and other aquatic organisms. This invariably affects humans by causing food insecurity, hunger and poverty. In an effort to curb this menace, this study investigated the effects of crude tuber peel extract of Tacca leontopetaliodes on the biochemical and histological parameters of Clarias gariepinus. Methods: Acute toxicity (96 hr . LC50) test was conducted after a range finding test from which definitive test concentrations of 5.00, 4.00, 3.00, 2.00, and 1.00 g/L were obtained with 0.00 g/L as control. One hundred and twenty (120) mixed sex C. gariepinus fingerlings (mean weight, 12.5 g ± 0.5; length, 13.2 cm ± 0.5) were administered definitive concentrations of crude tuber peel extract in duplicate replication. Water quality parameters (pH, dissolved oxygen, free carbon dioxide, alkalinity), tissue enzymes (Alkaline phosphatase (ALP), Alanine Aminotransferase (ALT), Aspartate Aminotransferase (AST) and histopathological assessments were analysed using standard procedures. Results: Physico-chemical parameters (pH, dissolved oxygen, free carbon dioxide and alkalinity) of test tanks containing graded concentrations of T. leontopetaliodes were significantly altered (p&lt;0.05) compared to the control tank. Similarly, the profile of hepatic enzymes (ALP, AST and ALT) displayed marked progressive dose-dependent increase levels relative to the control. Histopathological examination revealed graded deterioration of gills and liver architecture with increase in the strength of the extract T. leontopetaliodes Conclusion: The crude tuber peel extract of T. leontopetaliodes is toxic to fish, thus, washing, processing and discard of T. leontopetaliodes in and around riparian systems should be discouraged.

The regulatory effects of resveratrol on the expression of renal MMP-2 and MMP-9 in the rat models of diabetes.

Hyperglycemia caused by diabetes mellitus, in sensitive tissues such as the kidney, can cause dysfunction by damaging the blood vessels. Matrix metalloproteinases (MMPs), especially MMP-2 and MMP-9, play a significant role in the degradation of proteins and angiogenesis; while epidermal growth factor receptor (EGFR) is involved in the recovery of damaged tissues. Herein, we investigated the relationship between MMP-2 and MMP-9 / EGFR in the kidney tissues of streptozotocin-induced Wistar male rats. We also examined the effects of resveratrol known as a strong tissue-protective on these changes. Diabetes was induced by streptozotocin (55 mg/kg) and trans-resveratrol (20 mg/kg/day intraperitoneal) was used for treatment. Rats were divided into 4 groups as control, resveratrol (Res), diabetes (Diab) and diabetes plus resveratrol (Diab+Res). Superoxide dismutase (SOD) and catalase (CAT) levels were significantly decreased in the kidneys in the diabetic group, whereas nitrite / nitrate, urea, creatine kinase (CK) and uric acid concentrations increased. MMP-2, MMP-9, calcium-binding protein B (S100B) and platelet-derived growth factor (PDGF) concentrations of kidney were increased although reduced EGFR in the diabetes group. Resveratrol supplementation markedly restored all these structures. Diabetes activates MMP related inflammation and oxidative stress by suppressing antioxidant enzymes in the kidney tissues. Resveratrol has partly modulatory effects on diabetes-induced changes.

Matrix Metalloproteinases In Cancer Biology: A Review

Matrix metalloproteinases (MMPs) are a group of proteinases that are involved with the enzymatic breakdown of the extracellular matrix and basement membrane components. These enzymes are important in regulating vital physiological functions such as embryonic development, wound healing and tissue remodelling. However, several disorders may result from the exaggerated function of these enzymes such as ulcers, rheumatoid arthritis and invasive tumours. In tumours, the effect of these enzymes is not limited to invasion as it was traditionally believed but it also extends to the other oncogenic hallmark processes such as proliferation, angiogenesis, epithelial-mesenchymal transition and evasion from apoptosis. Therefore, it is essential to thoroughly understand the molecular mechanisms involved in these enzymes in cancerous tissue based on recent literature. Several reviews have highlighted the function of these enzymes in malignancies however the aim of this was to provide more recent overview to their role in malignant transformation and progression and in a briefer approach summarizing the complex molecular pathways. Online databases such as PubMed, Google scholar, Web of Science and MEDLINE were used to identify relevant articles. This approach would assist researchers by providing a list of the potential molecular targets in the sequence of changes related to these enzymes. This might help in designing a safer and a more specific targeted treatment for patients with cancer.

Simple isolation for two forms of malic enzyme from crustacean abdomen muscle

Abstract Two forms of NADP-dependent malic enzyme (ME, EC 1.1.1.40) were purified from the abdomen muscle of the crayfish Orconectes limosus (Rafinesque, 1817) and the shrimp Crangon crangon L., 1758 by affinity chromatography on 2′,5′-ADP-Sepharose 4B, with good qualitative recovery in a single step, using a substrate activation method with a malate–manganese chloride pair. The enzymes were identified by native polyacrylamide gel electrophoresis stained for protein and enzyme activity. The faster migrating mitochondrial enzyme from crayfish is inhibited by sulfhydryl reagent and loses its activity. Ellman’s Reagent, 5,5′-Dithiobis-(2-nitrobenzoic acid) (DTNB), can be used for the differentiation and measurement of cytoplasmic and mitochondrial malic enzyme in decapod crustacean tissue.

Effect of a Debaryomyces hansenii and Lactobacillus buchneri Starter Culture on Aspergillus westerdijkiae Ochratoxin A Production and Growth during the Manufacture of Short Seasoned Dry-Cured Ham.

Recently, specific dry-cured hams have started to be produced in San Daniele and Parma areas. The ingredients are similar to protected denomination of origin (PDO) produced in San Daniele or Parma areas, and include pork leg, coming from pigs bred in the Italian peninsula, salt and spices. However, these specific new products cannot be marked as a PDO, either San Daniele or Parma dry cured ham, because they are seasoned for 6 months, and the mark PDO is given only to products seasoned over 13 months. Consequently, these products are called short-seasoned dry-cured ham (SSDCH) and are not branded PDO. During their seasoning period, particularly from the first drying until the end of the seasoning period, many molds, including Eurotium spp. and Penicillium spp., can grow on the surface and work together with other molds and tissue enzymes to produce a unique aroma. Both of these strains typically predominate over other molds. However, molds producing ochratoxins, such as Aspergillus ochraceus and Penicillium nordicum, can simultaneously grow and produce ochratoxin A (OTA). Consequently, these dry-cured hams may represent a potential health risk for consumers. Recently, Aspergillus westerdijkiae has been isolated from SSDCHs, which could represent a potential problem for consumers. Therefore, the aim of this study was to inhibit A. westerdijkiae using Debaryomyces hansenii or Lactobacillus buchneri or a mix of both microorganisms. Six D. hansenii and six L. buchneri strains were tested in vitro for their ability to inhibit A. westerdijkiae. The strains D. hansenii (DIAL)1 and L. buchneri (Lb)4 demonstrated the highest inhibitory activity and were selected for in situ tests. The strains were inoculated or co-inoculated on fresh pork legs for SSDCH production with OTA-producing A. westerdijkiae prior to the first drying and seasoning. At the end of seasoning (six months), OTA was not detected in the SSDCH treated with both microorganisms and their combination. Because both strains did not adversely affect the SSDCH odor or flavor, the combination of these strains are proposed for use as starters to inhibit OTA-producing A. westerdijkiae.

Insulin Release from NPH Insulin-Loaded Pluronic® F127 Hydrogel in the Presence of Simulated Tissue Enzyme Activity

Background: Despite the widespread use of newer basal insulins, Natural Protamine Hagedorn (NPH) insulin still represents a well-established basal formulation with its long history of use, featuring the native form of human insulin. However, NPH insulin exhibits an undesirable peak within hours after a single subcutaneous (s.c.) injection, which may lead to hypoglycemia followed by insufficient basal insulin delivery. This may be attributed to the s.c. enzyme activities degrading the protamine in NPH microcrystals. Methods: A thermogelling block copolymer Pluronic® F127 (PF127) was utilized as a protective carrier for NPH microcrystals and as a modulator for insulin release from NPH. NPH insulin-loaded PF127 gel was prepared with varying concentrations of the polymer (15–25%) under mild conditions. The formulations were characterized for their gelling temperature, morphology, gel erosion, and in vitro insulin release, with trypsin concentrations up to 5 U/mL. Results: Scanning electron microscopy (SEM) showed that the integrity of NPH microcrystals was maintained after preparation. The burst release of insulin from NPH was significantly attenuated over the course of ~16h in the presence of PF127 with or without enzyme activity. Conclusion: NPH-PF127 successfully resisted the acceleration of NPH crystal dissolution and insulin release in vitro in the presence of protamine-degrading enzyme activity, warranting further testing.

Effect of Gas Flare on Some Clinical Enzyme Biomarkers of Eleme Residents in Rivers State, Nigeria

Background: Gas flaring, an environmental harmful practice, is prevalent in Eleme and most communities in the Niger Delta region of Nigeria.&#x0D; Aim: This study explored the impact of gas flaring on some clinical enzyme biomarkers of Eleme residents who are constantly exposed to the flared gas.&#x0D; Study Design/Place of Study: Volunteer subjects were randomly selected from representative groups resident in Eleme, an oil and gas producing and refining area, for over 15 years, while similar volunteer subjects resident in Nkpolu-Oroworukwo, a non-gas flaring community, served as the control.&#x0D; Methodology: Blood samples were collected from the subjects and analyzed for selected clinical enzyme biomarkers including Creatine kinase (CK), Lactate dehydrogenase (LDH) and Aspartate aminotransferase (AST) using standard enzyme activity and Randox test kit methods.&#x0D; Results: Results obtained revealed that CK levels for female Eleme subjects increased by 61.20% when compare with the female control subjects. This increase was statistically significant (p&lt;0.05). The CK levels for Eleme male subjects also increased by 37.36% compared to control. However, this increase was not statistically significant (p&lt;0.05). LDH increased by 23.21% in the male subjects and 18.58% in the female subjects compared to control, while AST increased by 42.11% in the male subject and 11.32% in the female subjects compared to control. The results suggest that there could be impending damage to organs for which an increase in the biomarkers-Creatine kinase, Lactate Dehydrogenase and Aspartate Aminotransferase suggest an ongoing pathologic process.&#x0D; Conclusion: These results therefore suggest that the continuous exposure to gas flare is causing an increase in some critical tissue and organ function enzyme biomarkers in blood. This may eventually affect the health status of the residents, increase the tendency of developing ill health and generally reduce their quality of life.

Human platelet lysate as a potential clinical-translatable supplement to support the neurotrophic properties of human adipose-derived stem cells

BackgroundThe autologous nerve graft, despite its donor site morbidity and unpredictable functional recovery, continues to be the gold standard in peripheral nerve repair. Rodent research studies have shown promising results with cell transplantation of human adipose-derived stem cells (hADSC) in a bioengineered conduit, as an alternative strategy for nerve regeneration. To achieve meaningful clinical translation, cell therapy must comply with biosafety. Cell extraction and expansion methods that use animal-derived products, including enzymatic adipose tissue dissociation and the use of fetal bovine serum (FBS) as a culture medium supplement, have the potential for transmission of zoonotic infectious and immunogenicity. Human-platelet-lysate (hPL) serum has been used in recent years in human cell expansion, showing reliability in clinical applications.MethodsWe investigated whether hADSC can be routinely isolated and cultured in a completely xenogeneic-free way (using hPL culture medium supplement and avoiding collagenase digestion) without altering their physiology and stem properties. Outcomes in terms of stem marker expression (CD105, CD90, CD73) and the osteocyte/adipocyte differentiation capacity were compared with classical collagenase digestion and FBS-supplemented hADSC expansion.ResultsWe found no significant differences between the two examined extraction and culture protocols in terms of cluster differentiation (CD) marker expression and stem cell plasticity, while hADSC in hPL showed a significantly higher proliferation rate when compared with the usual FBS-added medium. Considering the important key growth factors (particularly brain-derived growth factor (BDNF)) present in hPL, we investigated a possible neurogenic commitment of hADSC when cultured with hPL. Interestingly, hADSC cultured in hPL showed a statistically higher secretion of neurotrophic factors BDNF, glial cell-derived growth factor (GDNF), and nerve-derived growth factor (NFG) than FBS-cultured cells. When cocultured in the presence of primary neurons, hADSC which had been grown under hPL supplementation, showed significantly enhanced neurotrophic properties.ConclusionsThe hPL-supplement medium could improve cell proliferation and neurotropism while maintaining stable cell properties, showing effectiveness in clinical translation and significant potential in peripheral nerve research.

Journal-related Activities and Other Special Activities at the 2020 American Society of Anesthesiologists Meeting.

Journal-related Activities and Other Special Activities at the 2020 American Society of Anesthesiologists Meeting.

Effect of Aspergillus flavus on lipid peroxidation and activity of antioxidant enzymes in midgut tissue of Spodoptera litura larvae

In the present study, effect of Aspergillus flavus was analysed on lipid peroxidation and activities of antioxidant enzymes in midgut of Spodoptera litura. Fungus was administered to the larvae of S. litura using artificial diet method having 1.34 mg/mL (LC50) concentration. The fungus toxicity was observed after 24, 48, 72 and 96 h post treatment in the midgut tissue of insect larvae by examining lipid peroxidation (MDA content) and activities of antioxidant enzymes viz superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APOX) and glutathione-S-transferase (GST) as oxidative stress markers. Additionally, repair of the damage caused due to fungal treatment was also assessed to decipher the longevity of effect by giving control diet to the exposed larvae for two days after particular time exposure using same parameters. In particular, significant (p ≤ 0.05) increased level of lipid peroxidation and antioxidant enzymes activities was observed in all exposure time groups with respect to control which showed that midgut of exposed larvae has oxidative predominance. Repair studies demonstrated the negligible repair in insect larvae. In conclusion, A. flavus contributed to strong oxidative stress and toxicity in S. litura larvae.

Alterations in Enzyme Activity of Carbonic Anhydrase, 6-phosphogluconate Dehydrogenase and Thioredoxin Reductase in Rats Exposed to Doxorubicin and Morin

Objective: Carbonic anhydrase (CA), 6-phosphogluconate dehydrogenase (6PGD) and thioredoxin reductase (TrxR) enzymes are the essential biological molecules needed for metabolic processes in all living cells. This study was designed to investigate the activities of CA, 6PGD and TrxR enzymes in the brain, kidney, liver, heart and testis tissues of the rats exposed to Dox and morin. Methods: Male Wistar albino rats were randomly divided into three groups as control, morin and DOX, each of them containing 7 rats. At the end of the experimental procedure, CA, 6PGD and TrxR enzyme activities in tissues of rats were determined spectrophotometrically. Results: In our study, we observed that DOX activated CA enzyme in liver and kidney tissues while inhibiting CA enzyme in the other tissues, activated 6PGD enzyme in the kidney, liver and heart tissues, and inhibited the TrxR enzyme in all the tissues. In addition, morin activated CA enzyme in the liver tissue while inhibiting CA enzyme in the brain, heart and testis tissues. Morin activated 6PGD enzyme activity while it inhibited TrxR enzyme in all the tissues. Conclusion: The findings showed that doxorubicin and morin had similar properties in the tissues as to their effect on enzyme activities.