Isoenzyme Patterns Research Articles

Biochemical Changes in Embryogenic and Nonembryonic Callus of Glycyrrhiza glabra L. during Somatic Embryogenesis

The continuous extraction of glycyrrhizin from Glycyrrhiza glabra L. has led to the critical endangerment of this therapeutic plant. Glycyrrhizin, a triterpenoid saponin, is a vital secondary metabolite renowned for its diverse pharmacological benefits, including anti-inflammatory, immunomodulatory, antiallergic, antiulcer properties, and even efficacy against HIV. However, the relentless harvesting of roots for high-value glycyrrhizin results in the obliteration of the entire plant, posing a severe threat to its existence and causing a detrimental impact on biodiversity. To counteract this challenge and conserve the species, sustainable approaches are imperative. One such approach involves the utilization of in vitro regeneration through somatic embryogenesis for clonal proliferation and genetic modification. This report presents findings on SE-associated proteins in Glycyrrhiza glabra L., accomplished through the implementation of the discontinuous gel electrophoresis technique. The study focused on a comparative isozyme analysis employing non-embryogenic and embryogenic calli of G. glabra L. Remarkably, these isozymes were exclusively identified and exhibited affiliations with a spectrum of cellular functions. A substantial portion of these proteins are engaged in a diverse range of metabolic and stress-related functions. This study utilized crude enzyme/total protein extracts as a basis to evaluate the genetic stability of in vitro cultivated plant tissue. Acid phosphatase, esterase, and peroxidase were selected as indicators for this assessment. Intriguingly, esterase and acid phosphatase exhibited significant polymorphism between the segregated embryogenic callus (EC) and non-embryogenic callus (NEC) derived from the same G. glabra L. genotype. Comparative analyses encompassing esterase, acid phosphatase, and peroxidase activities, alongside the expression profiles of these isozymes throughout somatic embryogenesis (SE), were conducted using polyacrylamide gel electrophoresis. Notably, the isoenzyme patterns of calluses were assessed. The activity of isozymes within an embryogenic line of callus surpassed that within a non-embryogenic counterpart. However, no conspicuous alterations in quality were observed between the non-embryogenic and embryogenic calluses.

Establishment of Big Bone chicken fibroblast cell bank and study of its biological characteristics

A fibroblast bank of Big Bone chicken was established using tissue adherent culture method. This cell bank included 29 embryo samples, had stocks of 147 cryogenically-preserved vials each containing 2~3Å~106 cells, and met all the cell line quality control standards established by the American Type Culture Collection (ATCC). The cells cultured in vitro showed the typical morphology of fibroblasts. The growth curve assumed the “S” shape, and the cell population doubling time (PDT) was about 30h. Cell viability was 98.1% before cryopreservation and 96.6% after recovery. All tests for microbial contamination were negative. The isoenzyme pattern was of species specificity. The frequency of diploid cells was 91%. The transfection efficiency of three fluorescent protein genes fluctuated between 10.6% and 26.5%. These results showed that the cells cultured in vitro grew well and had stable genetic properties. This research thus does not only preserve the poultry genetic resources of Big Bone chicken at the cell level, but also opens new ways for preserving important genetic resources of endangered animals in the form of somatic cells.

Isoenzyme characterization of Leishmania infantum toward checking the antioxidant activity of superoxide dismutase and glutathione peroxidase

BackgroundLeishmania infantum is the major causative agent of visceral leishmaniasis in Mediterranean regions. Isoenzyme electrophoresis (IE), as a biochemical technique, is applied in the characterization of Leishmania species. The current study attempted to investigate the isoenzyme patterns of logarithmic and stationary promastigotes and axenic amastigotes (amastigote-like) of L. infantum using IE. The antioxidant activity of superoxide dismutase (SOD) and glutathione peroxidase (GPX) was also checked in the aforementioned forms.MethodAfter L. infantum cultivation and obtaining logarithmic and stationary promastigotes, axenic amastigotes were achieved by incubation of stationary promastigotes at 37 °C for 48 h. The lysate samples were prepared and examined for six enzymatic systems including glucose-6-phosphate dehydrogenase (G6PD), nucleoside hydrolase 1 (NH1), malate dehydrogenase (MDH), glucose-phosphate isomerase (GPI), malic enzyme (ME), and phosphoglucomutase (PGM). Additionally, the antioxidant activity of SOD and GPX was measured.ResultsGPI, MDH, NH1, and G6PD enzymatic systems represented different patterns in logarithmic and stationary promastigotes and axenic amastigotes of L. infantum. PGM and ME showed similar patterns in the aforementioned forms of parasite. The highest level of SOD activity was determined in the axenic amastigote form and GPX activity was not detected in different forms of L. infantum.ConclusionThe characterization of leishmanial-isoenzyme patterns and the measurement of antioxidant activity of crucial antioxidant enzymes, including SOD and GPX, might reveal more information in the biology, pathogenicity, and metabolic pathways of Leishmania parasites and consequently drive to designing novel therapeutic strategies in leishmaniasis treatment.

The hepato- and neuroprotective effect of gold Casuarina equisetifolia bark nano-extract against Chlorpyrifos-induced toxicity in rats

BackgroundThe bark of Casuarina equisetifolia contains several active phytoconstituents that are suitable for the biosynthesis of gold nanoparticles (Au-NPs). These nanoparticles were subsequently evaluated for their effectiveness in reducing the toxicity induced by Chlorpyrifos (CPF) in rats. ResultsVarious hematological and biochemical measurements were conducted in this study. In addition, markers of oxidative stress and inflammatory reactions quantified in liver and brain tissues were evaluated. Histopathological examinations were performed on both liver and brain tissues. Furthermore, the native electrophoretic protein and isoenzyme patterns were analyzed, and the relative expression levels of apoptotic genes in these tissues were determined. The hematological and biochemical parameters were found to be severely altered in the group injected with CPF. However, the administration of Au-C. equisetifolia nano-extract normalized these levels in all treated groups. The antioxidant system markers showed a significant decrease (P ≤ 0.05) in conjunction with elevated levels of inflammatory and fibrotic markers in both liver and brain tissues of the CPF-injected group. In comparison, the pre-treated group exhibited a reduction in these markers when treated with the nano-extract, as opposed to the CPF-injected group. Additionally, the nano-extract mitigated the severity of histopathological lesions induced by CPF in both liver and brain tissues, with a higher ameliorative effect observed in the pre-treated group. Electrophoretic assays conducted on liver and brain tissues revealed that the nano-extract prevented the qualitative changes induced by CPF in the pre-treated group. Furthermore, the molecular assay demonstrated a significant increase in the relative expression of apoptotic genes in the CPF-injected rats. Although the nano-extract ameliorated the relative expression of these genes compared to the CPF-injected group, it was unable to restore their values to normal levels. ConclusionOur results demonstrated that the nano-extract effectively reduced the toxicity induced by CPF in rats at hematological, biochemical, histopathological, physiological, and molecular levels, in the group pre-treated with the nano-extract.

Isoenzymatic Pattern of Hydrogen Sulfide (H2S)-Generating L-Cysteine Desulfhydrase (LCD) in Arabidopsis thaliana Seedlings: Effect of Nitric Oxide (NO) and H2S

In higher plants, hydrogen sulfide (H2S) is a recognized signaling molecule that performs multiple regulatory functions. The enzyme L-cysteine desulfhydrase (LCD) catalyzes the conversion of L-cysteine (L-Cys) to pyruvate and ammonium with the concomitant generation of H₂S, and it is considered one of the main sources of H2S in plants. Using non-denaturing polyacrylamide gel electrophoresis (PAGE) in combination with a specific assay for LCD activity, this study aims to identify the potential LCD isozymes in wild-type Arabidopsis thaliana seedlings of 16 days old grown under in vitro conditions, and to evaluate the potential impact of nitric oxide (NO) and H2S on these LCD isozymes. For this purpose, an Atnoa1 mutant characterized to have a low endogenous NO content as well as the exogenous application of H2S were used. Five LCD isozymes were detected, with LCD IV being the isozyme that has the highest activity. However, the LCD V activity was the only one that was positively modulated in the Atnoa1 mutants and by exogenous H2S. To our knowledge, this is the first report showing the different LCD isozymes present in Arabidopsis seedlings and how their activity is affected by NO and H2S content.

Comparison of Isoenzyme Pattern of Echinococcus granulosus sensu stricto (G1-G3) and E. canadensis (G6/G7) Protoscoleces

Different genotypes of Echinococcus granulosus sensu lato (s.l.) infect humans and ungulate animals, causing cystic echinococcosis. Simultaneous isoenzyme, as well as molecular characterizations of this parasite, has not yet been investigated in Iran. The present study aimed to evaluate the isoenzyme pattern of the E. granulosus sensu stricto (s.s.) and E. canadensis genotypes in Iran. A total of 32 (8 humans and 24 animals) cystic echinococcosis cysts were isolated from Shiraz, Tehran, Ilam, and Birjand from May 2018 to December 2020. The DNAs were extracted and their genotypes were determined by molecular methods. Enzymes were extracted from the cysts and subjected to polyacrylamide gel electrophoresis. The activities of glucose-6-phosphate sehydrogenase (G6PD), malate dehydrogenase (MDH), malic enzyme (ME), nucleoside hydrolyse 1 (NH1), and isocitrate dehydrogenase (ICD) were examined in the cyst samples using isoenzyme method and compared it with the genotyping findings. DNA sequence analysis of the samples showed that the specimens contained 75% E. granulosus s.s. (G1) and 25% E. canadensis (G6) genotypes. The isoenzyme pattern of ICD in both genotypes produced a six-band pattern with different relative factors. The G6PD also produced two bands with different relative migrations in both genotypes. The MDH and NH1 systems revealed a two-band pattern, while only one band was generated in the ME enzyme in the E. granulosus s.s. genotype. In the E. canadensis, the MDH and NH1 enzymes showed one band, and the ME enzyme represented a two-band pattern. Our findings suggest that E. granulosus s.s. and E. canadensis genotypes have entirely different isoenzyme patterns for NH1, G6PD, MDH, and ME.

Elevation of Lactate Dehydrogenase & its Isoenzyme Pattern in Pregnancy induced Hypertension: Etiopathogenesis and Consequences

Lactate dehydrogenase (LDH) is an intra cellular cytosolic enzyme, which play an important role in anaerobic Glycolysis. It is a tetramer having four subunits which may be all H type or all M type or it may be a mix of both these subunits. Preeclampsia is defined as a pregnancy-specific multi-systemic syndrome of widespread endothelial malfunction and vasospasm developing after 20 weeks of gestation. Analysis of Isoenzyme pattern of LDH is more important than Total LDH value, anaerobic form LDH is seen to be predominantly found in case of pre-eclampsia.

Evaluating the typing power of six isoenzymatic systems for differentiation of clinical and standard isolates of Candida species

Background: Due to the increasing prevalence of candidiasis, early detection of the causative agents may pave the way for the management of this infection. The present study aimed to assess the discriminative power of the six isoenzymatic systems for differentiating the Candida species.Materials and Methods: Sixteen standard Candida albicans and Candida dubliniensis strains and 30 fluconazole-sensitive and fluconazole-resistant clinical strains of Candida albicans were analyzed using a Multilocus Enzyme Electrophoresis (MLEE) method, including six enzymatic systems consisting of malate dehydrogenase (MDH), phosphoglucomutase (PGM), glucose-phosphate isomerase (GPI), glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGD), and malic enzyme (ME).Results: Among the six enzymatic systems, ME showed no diagnostic activity, whereas MDH provided the best species-specific pattern for species discrimination. In addition, the MDH and G6PD systems provided a discriminatory pattern for differentiating C. dubliniensis from C. albicans isolates. The same isoenzymatic activity was detected in all 36 standard and clinical isolates. Moreover, the results showed no correlation between the isoenzymatic profiles and drug resistance.Conclusion: Among the investigated MLEE systems, MDH was able to differentiate between Candida albicans and Candida dubliniensis. Although no association was detected between isoenzyme patterns and fluconazole resistance in this investigation, isoenzyme patterns are likely correlated with virulence factors between species and even within species. To answer these questions, additional studies should be done on more strains.

Agarose gel electrophoresis pattern of serum creatine kinase and lactate dehydrogenase isoenzymes in zoo-managed Asian elephants (Elephas maximus).

Serum levels of creatine kinase (CK) and lactate dehydrogenase (LDH) isoenzymes were evaluated in nine zoo-managed Asian elephants (Elephas maximus) using a commercial agarose gel electrophoresis (AGE) kit. CK was separated into two major fractions, CK-BB and CK-MM, along with a small fraction of macroenzyme-CK type 2 (mCK2); CK-MM was the largest fraction. LDH was separated into five fractions (LDH1-5); LDH3 was the largest fraction. Age was negatively and positively correlated with the percentages of CK-BB and CK-MM, respectively, and negatively correlated with CK-BB and mCK2 activities. These results indicate that an AGE kit can be used to evaluate CK and LDH isoenzymes. Routine isoenzyme testing may enable early detection of disease and physiological changes.

The impaired distribution of adenosine deaminase isoenzymes in multiple sclerosis plasma and cerebrospinal fluid.

BackgroundAdenosine deaminase (ADA) via two isoenzymes, ADA1 and ADA2, regulates intra- and extracellular adenosine concentrations by converting it to inosine. In the central nervous system (CNS), adenosine modulates the processes of neuroinflammation and demyelination that together play a critical role in the pathophysiology of multiple sclerosis (MS). Except for their catalytic activities, ADA isoenzymes display extra-enzymatic properties acting as an adhesion molecule or a growth factor.AimsThis study aimed to explore the distribution and activity of ADA1 and ADA2 in the plasma and the CSF of MS patients as well as in the human brain microvascular endothelial cells (HBMEC), human brain vascular pericytes and human astrocytes.Methods and resultsThe enzyme assay following reverse phase-high performance liquid chromatography (HPLC) analysis was used to detect the ADA1 and ADA2 activities and revealed an increased ratio of ADA1 to ADA2 in both the plasma and the CSF of MS patients. Plasma ADA1 activity was significantly induced in MS, while ADA2 was decreased in the CSF, but significance was not reached. The brain astrocytes, pericytes and endothelial cells revealed on their surface the activity of ADA1, with its basal level being five times higher in the endothelial cells than in the astrocytes or the pericytes. In turn, ADA2 activity was only observed in pericytes and endothelial cells. Stimulation of the cells with pro-inflammatory cytokines TNFα/IL17 for 18 h decreased intracellular nucleotide levels measured by HPLC only in pericytes. The treatment with TNFα/IL17 did not modulate cell-surface ATP and AMP hydrolysis nor adenosine deamination in pericytes or astrocytes. Whereas in endothelial cells it downregulated AMP hydrolysis and ADA2 activity and upregulated the ADA1, which reflects the ADA isoenzyme pattern observed here in the CSF of MS patients.ConclusionIn this study, we determined the impaired distribution of both ADA isoenzymes in the plasma and the CSF of patients with MS. The increased ADA1 to ADA2 ratio in the CSF and plasma may translate to unfavorable phenotype that triggers ADA1-mediated pro-inflammatory mechanisms and decreases ADA2-dependent neuroprotective and growth-promoting effects in MS.

Isoenzyme study of causative agents of leishmaniasis using glutamate dehydrogenase and hexokinase

Leishmania Spp. are a group of protozoan parasites which cause a wide range of human diseases from localized self-healing cutaneous lesions to fatal visceral infections. Isoenzyme characterization by using various enzyme systems is a reliable and important tool for differentiation between various species of protozoa especially family trypanosomatidae including genus Leishmania. In this study two enzyme systems were evaluated for identification of isolated organisms from patients with cutaneous leishmaniasis (CL) in endemic foci of Fars province South of Iran. In some cases, it was observed that some enzyme systems routinely used in Leishmania parasite identification centers do not differentiate well between species, so it was decided to use non routine enzyme systems. Isoenzyme pattern of two enzyme systems, Gloutamate Dehydrogenase (GDH) and Hexokinase (HK) were compared by Poly Acryl amide Gel Electrophoresis (PAGE) and Cellulose acetate Electrophoresis (CE) in characterization of Leishmania major, Leishmania tropica and Leishmania infantum. Our findings showed the enzymes GDH on PAGE could differentiate L. major from L. tropica and L. infantum. The HK enzymatic system used on CE in this study has ability to differentiate three species of L. major, L. tropica and L. infantum.

Antioxidant Defense during Recovery of Resurrection Plant Haberlea rhodopensis from Drought- and Freezing-Induced Desiccation.

In this study, the contribution of nonenzymatic (ascorbate, glutathione) and enzymatic antioxidants (superoxide dismutase, catalase, glutathione reductase, glutathione S-transferase) in the first hours of recovery of the resurrection plant Haberlea rhodopensis from drought- and freezing-induced desiccation was assessed. The initial stage of recovery after desiccation is critical for plants, but less investigated. To better understand the alterations in the activity of antioxidant enzymes, their isoenzyme patterns were determined. Our results showed that ascorbate content remained high during the first 9 h of rehydration of desiccated plants and declined when the leaves′ water content significantly increased. The glutathione content remained high at the first hour of rehydration and then strongly decreased. The changes in ascorbate and glutathione content during recovery from drought- and freezing-induced desiccation showed great similarity. At the beginning of rehydration (1–5 h), the activities of antioxidant enzymes were significantly increased or remained as in dry plants. During 7–24 h of rehydration, certain differences in the enzymatic responses between the two plant groups were registered. The maintenance of a high antioxidant activity and upregulation of individual enzyme isoforms indicated their essential role in protecting plants from oxidative damage during the onset of recovery.

Antioxidative enzymes in root and leaf of Rumex obtusifolius L. grown on ash amended soil

Antioxidant enzymes are one of the most important links in the plant defense system to various types of environmental stress, so their response to a particular type of stress may indicate the sensitivity or tolerance of the plant species. Our paper studied the difference in antioxidative enzyme [catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) and Class III peroxidases (POD, EC 1.11.1.7)], isoenzyme pattern and activities between Rumex obtusifolius L. (Polygonaceae) plants grown on ash amended and uncontaminated soil. Modified SDS-PAGE electrophoresis revealed the presence of a new POD isoform in leaf samples growing on ash-amended soil, although the activity of POD in the leaves did not change significantly compared to control plants. On the other hand, in the roots of ash-growing plants POD activity decreased by 90%. Single CAT isoform was detected in both leaf samples, and results indicate 47% higher CAT activity in leaves of ash growing plants. Native electrophoresis detected two SOD isoforms in leaves and roots from the control plant. SOD isoforms were inhibited in the roots of plants grown on ash. The paper indicates the possible role of CAT, SOD and POD in the adaptive response of R. obtusifolius plants on ash amended soil.

Assessment of the Anti-Cancer Efficiency of Silver Moringa oleifera Leaves Nano-extract against Colon Cancer Induced Chemically in Rats.

Background:Colorectal cancer (CRC) categorized as the most common type of gastrointestinal cancers affected both genders equally. Chemotherapeutic drugs became limited due to their deleterious side effects. Therefore, efficiency of M. oleifera leaves extract increased by incorporating silver nanoparticles (Ag-NPs) then studied against colon cancer induced by azoxymethane (AOM) in rats. Methods:Different hematological and biochemical measurements in addition to specific tumor and inflammatory markers were quantified. Histopathological examination for Colonic tissues was performed. Native proteins and isoenzyme patterns were electrophoretically detected in addition to assaying expression of Tumor Protein P53 (TP53) and Adenomatous Polyposis Coli (APC) genes in colonic tissues.Results: M. oleifera nano-extract restored levels of the hematological and biochemical measurements in addition to levels of tumor and inflammatory markers to normalcy in both of nano-extract simult- and post-treated groups. Also, it minimized severity of the histopathological alterations in the simult-treated group and prevented it completely in the post-treated group. The lowest similarity index (SI%) values were noticed with electrophoretic protein (SI=61.54%), lipid (SI=0.00%) and calcium (SI=75.00%) moieties of protein patterns, catalase (SI=85.71%), peroxidase (SI=85.71%), α-esterase (SI=50.00%) and β-esterase (SI=50.00%) isoenzymes in addition to altering the relative quantities of total protein and isoenzyme bands in colon of cancer induced group. Moreover, levels of TP53 and APC gene expression increased significantly (P≤0.05) in colon cancer induced group. The nano-extract prevented the qualitative and quantitative alterations in the different electrophoretic patterns in addition to restoring levels of the gene expressions to normalcy in both of simult- and post-treated groups. Conclusion: M. oleifera nano-extract exhibited ameliorative effect against the biochemical, physiological and molecular alterations induced by AOM in nano-extract simult- and post-treated groups.

A Study of Arginase Expression in Chronic Non-healing Wounds.

Arginase expression has been recently shown to increase in numerous disease states like neurodegeneration, inflammation, and malignancies. Although it has been found to be functionally important in various disease pathologies, little is known about its role in wound healing. Here, we look at the expression of arginase and its isoforms in chronic non-healing wounds and also study the expression of nitric oxide synthase (NOS) and oxidative stress enzymes in them. Wound tissues and blood samples were collected at the time of index presentation and follow-up from 61 chronic non-healing wound cases. The expression patterns of arginase isoenzymes, NOS, superoxide dismutases (SOD), lactic acid dehydrogenase (LDH), and catalase were examined by using enzyme-linked immunosorbent assay, immunohistochemistry, and western blot analysis at the transcript and protein level. We reported a significant decrease of serum arginase levels in chronic nonhealing wounds in the progress of wound healing. Interestingly, tissue arginase levels were found to be increased with improved wound condition at follow-up. Tissue NOS, LDH, and catalase activity were also found to be increased with the progress of healing, whereas SOD levels were downregulated. Our findings reported increased expression at the transcript level of arginase-I and arginase-II in chronic non-healing wounds for the first time. In conclusion, we observed decreased serum arginase levels in completely healed patients as compared to non-healed cases. Our study findings support the hypothesis that inhibition of the activity of arginase delays wound healing. Arginase and iNOS may also find their place in the future as possible biomarkers for wound healing.

Diverse activities and biochemical properties of amylase and proteases from six freshwater fish species

This study investigated the biochemical properties, enzyme activities, isoenzyme pattern, and molecular weight of three types of digestive enzyme from six freshwater fish species: Puntius gonionotus (common silver barb), Puntioplites proctozysron (Smith’s barb), Oreochromis niloticus (Nile tilapia), Hemibagrus spilopterus (yellow mystus), Ompok bimaculatus (butter catfish), and Kryptopterus geminus (sheatfish). The optimum pHs for amylase and alkaline protease activities were 7.0–8.0 and 8.0–10.0, and the optimum temperatures were 45–60 °C and 50–55 °C, respectively. A pepsin-like enzyme was detected in all three carnivorous fishes (Ompok bimaculatus, Kryptopterus geminus, and Hemibagrus spilopterus) with optimum reaction pH of 2.0 for each and optimum reaction temperatures 50–55 °C. In optimum reaction conditions, the amylase and alkaline protease from Puntioplites proctozyron showed the highest activities. Lower activities of all enzymes were observed at temperature (29 °C) of Lam Nam Choen swamp than at the optimum reaction temperatures. The fish species contained one to three and five to eight isoforms of amylase and alkaline protease, respectively, with molecular weights from 19.5 to 175 kDa. Both the alkaline proteases and amylases were stable in wide pH and temperature ranges.

Isoenzyme Characterization of Trichomonas vaginalis Isolated from HIV Patients in Fars and Kerman, Southeast Iran

Background: Trichomonas vaginalis is an anaerobic flagellated protozoan which is responsible for human urogenital infections. Several zymodemes of T. vaginalis have been reported from various parts of the worlds on the basis of isoenzyme patterns. This study was conducted to characterize the isolated organisms of T. vaginalis from HIV patients using isoenzyme electrophoresis in Fars and Kerman provinces, southeast Iran. Methods: Eighteen mass cultivated isolates of T. vaginalis in the modified TYI-S-33 medium were analyzed using isoenzyme electrophoresis. Polyacrylamide gel electrophoresis (PAGE) of five different enzyme systems were used to characterize T. vaginalis isolates: (i) Glucose-6-phosphate dehydrogenase (G6PD), (ii) Glucose phosphate isomerase (GPI), (iii) Malate dehydrogenase (MDH), (iv) Malic enzyme (ME), and (v) Phosphoglucomutase (PGM). Results: MDH, GPI, PGM, and ME enzyme systems showed a homogeneity and detected an identical enzyme pattern in all isolates. Meanwhile, G6PD revealed two different enzyme patterns. The isoenzyme electrophoretic profiles divided 18 T. vaginalis isolates into two zymodemes. Zymodeme 1 contained Shiraz isolates and zymodeme 2 contained Kerman isolates. Conclusion:The polymorphism of Iranian human isolates of T. vaginalis could be assessed by biochemical study using appropriate enzyme systems. Isoenzyme analysis is a promising method for the characterization of T. vaginalis. New molecular studies with increased number of enzyme loci and genetic markers are suggested to classify more zymodemes of Trichomonas in Iran.

Evaluation of the Hepato- and Neuroprotective Effect of Chemical Constituents in Saussurea costus Extract Against the Toxicity Induced by Chloropyrofos Ethyl in Rats

The health hazardous occurred in both human and animals as a result of the continuous misuse of pesticides drives the researchers for looking about the solution. Natural plant extracts contain active phyto-constituents with antioxidant potency enables them to inhibit production of the free radicals that induce damage of the cells. The present study aimed to reveal efficiency of Saussurea costus (S. costus) extract against the hepato- and neurotoxicity induced by Chloropyrifos ethyl (CPF) in experimental animals (rats). The gas chromatography / mass spectrometer (GC/MS) that used for analyzing the active constituents in S. costus extract showed that the extract contains 11 potent active compounds and Dehydrocostuslactone represents about 77.37% and considered as the most dominant compound in the extract. Both of CPF and S. costus extract were studied on the rats that were divided into 6 groups as the following: Group 1 (control) received distilled water orally. Groups 2&3 (S. costus extract treated groups) received S. costus extract orally at a dose of 0.25 and 0.50 ml, respectively. Group 4 (CPF injected group) was injected with CPF at a dose of 3 mg/kg.bw. Groups 5&6 (CPF + S. costus extract group) injected with CPF then treated with S. costus extract at two tested doses respectively. The most hematological and biochemical measurements declined significantly (P≤0.05) in CPF injected group. S. costus extract restored all tested parameters towards the control values. Moreover, the electrophoretic isoenzyme showed that the physiological alterations occurred in the esterases (ESTs) as a result of CPF injection were represented by hiding normal EST types associated with existence of abnormal ones. Therefore, the similarity index (SI%) and genetic distance (GD%) values were altered with α-EST (SI=80.00%; GD=20.00%) and β-EST (SI=50.00%; GD=50.00%) patterns in CPF injected group. The S. costus extract at a dose of 0.5 ml restored integrity of these isoenzymes pattern by restoring the absent types with hiding the abnormal ones. Therefore, this group became physiologically similar to control group (SI=100.00%; GD=0.00%). These results were supported by histopathological examination for the target organs (brain, liver and kidney) that were affected by CPF and the S. costus extract improved architecture of these organs and restored their histopathological integrity to normal structure.

Comparative Study between Croton tiglium Seeds and Moringa oleifera Leaves Extracts, after Incorporating Silver Nanoparticles, on Murine Brains

Croton tiglium seeds and Moringa oleifera leaves extracts are rich in phytoconstituents with the antioxidant efficiency which can be enhanced by incorporating silver nanoparticles (Ag-NPs). The present study was designed to compare the effect of C. tiglium seeds and M. oleifera leaves nano-extracts on brain tissues of murine models. During the current study, acetylcholine esterase (ACHE), β-amyloid (Aβ) content and inflammatory markers were measured in brain tissues. Moreover, native protein, lipoprotein and isoenzymes patterns were electrophoretically detected. Also, the interferon-gamma (INF-γ) receptor protein was studied by molecular dynamic simulation to evaluate the significant alterations on brain tissues. It was found that ACHE, Aβ contents and inflammatory markers increased in the C. tiglium nano-extract treated group at a dose of 6.5 ml/kg. Furthermore, it caused qualitative electrophoretic abnormalities represented by lowering similarity index (SI) values. Also, the resides range 119~127 represent the most reactive and flexible site in INF-γ receptor protein. On the other hand, it was shown that no significant differences were induced by silver M. oleifera nano-extract.

LACTATE DEHYDROGENASE ISOENZYMES IN TISSUES OF CARNIVOROUS MAMMALS DURING POSTNATAL ONTOGENY

The electrophoretic separation of isoenzymes of lactate dehydrogenase (LDH, EC 1.1.1.27) was performed in the kidneys, skeletal muscle, lungs and spleen in mink Neovison vison Schreber and blue fox Alopex lagopus L. pups at 15, 25, 60, 90, 120, 150, 180, and 210 days of age. Significant changes in the isoenzyme spectra of LDH in the tissues of mink and blue fox pups were observed in the initial period of the animals’ intensive growth and development –at the age of 15–0 days. In the skeletal muscles, lungs and spleen, the content of M-subunits of LDH during postnatal ontogeny was higher in mink than in foxes. The isozyme profile of renal LDH in 4‑onth-old pups of mink and blue foxes corresponded to that of 7‑onth-old animals. The tissue specificity of the LDH isoenzyme pattern reflects the metabolic profile of the tissues and is detectable already in early developmental stages.