Esterase Bands Research Articles

Esterase Inhibition Study in Different Tissues (Intestine, Muscle and Brain) of Fresh Water Cat Fish Heteropneustesfossilis through Polyacrylamide Gel Electrophoresis

Esterase polymorphism was studied in different tissues of stinging catfish (Heteropneustesfossilis) viz; intestine, muscle and brain. Intestine showed ER esterases(Esterases resistant to inhibitors) and CE esterases (Carboxylesterases), muscle showed ChE (Cholinesterases) and CHspesterases (Enzymes which were inhibited by Paraoxon, Eserine, and pCMB) and brain showed ChE(Cholinesterases) and Esdp(Esterases inhibited by Eserine alone) esterases. The present study was carried out to find out tissue specific esterase banding patterns in different tissues of H.fossilis. Electrophoretic banding patterns of esterase of different tissues showed species specific variation which could be successfully used for identification of fish species. Electrophoretic pattern of esterases of different tissues show species specific variation, it could be successfully used for the identification of fish species

Population study of Cucurbita moschata based on morphological characters and isozyme banding patterns in Bima District, West Nusa Tenggara, Indonesia

Abstract. Furqan M, Suranto, Sugiyarto. 2023. Population study of Cucurbita moschata based on morphological characters and isozyme banding patterns in Bima District, West Nusa Tenggara, Indonesia. Biodiversitas 25: 30-38. Pumpkin is a tropical plant that benefits from its lipid, protein, carbohydrate, and mineral contents. The morphological characters of pumpkins can be influenced by their genetics and environment. This research aimed to examine the relationship among 9 accessions of Cucurbita moschata Duchesne from 9 locations in Bima district, West Nusa Tenggara (NTB) province, Indonesia. The morphological characters of stems, leaves, and flowers were analyzed qualitatively and quantitatively and an ANOVA was used to look at the difference of leaf and flower characters. Polyacrylamide gel electrophoresis (PAGE) with peroxidase and esterase was used to detect isozyme banding patterns. Morphological characters and isozyme data were analyzed using a UPGMA method with NTSYS version 2.0 software to construct a dendrogram. The results show evidence of the varied morphological appearance of C. moschata from the different locations. A total of 12 esterase bands and 10 peroxidase bands were detected from Wera2. An undetected band with Rf value of 0.567 was recorded, while a unique band with Rf value of 0.133 (peroxidase) was not detected. Undetected bands with Rf values of 0.645 and 0.726 of esterase were also recorded. A dendrogram showed three clusters of 9 accessions studied, in which Wera2 and Wera3 each stood alone, and a third large group was formed from the 7 remaining accessions. These results confirm the uniqueness of the Wera2 and Wera3 accessions. These results would be good information in determining a management strategy for improving the quality of pumpkins in the future.

Effect of Malathion (An Organophospahte) on Electrophoretic Banding Patterns of Esterase Isozymes in Gill, Liver, Brain Tissue of Fresh Water Fish Channa Punctatus (Bloch)

This investigation was conducted to compare the electrophoretetic banding patterns of esterase isozyme in the gill, liver, and brain tissues of the freshwater fish Channa punctatus (Bloch) at 24 hours, 48 hours, 72 hours, and 96 hours after exposure to Malathion (an Organophosphate) to those at the control group. Quantitative analysis of the esterase isozymes was performed using 7.5 native polyacrylamide gel electrophoresis (PAGE) stained with -naphthyl acetate as substrate. The relative mobilities of three esterase Isozyme bands in gill, liver, and brain tissue were determined to be 0.60.05, 0.40.05, and 0.30.05, respectively; these bands were designated Est-1, Est-2, and Est-3, respectively. Control samples from the gill, liver, and brain all had all three esterase bands. Both Est 2 and Est-3 Esterase Isozyme bands in gill and liver tissue were eliminated when fish were subjected to Malathion (an Organophosphate) for 72 and 96 hours, respectively. Malathion induced greater damage in Est-1 and Est-3 brain tissue than in Est-2.

Urine Dipstick Leukocyte Esterase in the Rapid Diagnosis of Septic Arthritis.

This study aimed to evaluate the sensitivity and specificity of the leukocyte esterase (LE) band in two groups of patients receiving and not receiving antibiotics and compare the results. This prospective cross-sectional study was conducted on 105 joints with clinical suspicion of infectious arthritis (based on Kocher criteria) admitted in Shohada Hospital, Tabriz, Iran, within 2017-2018. Patients were divided into two groups, including receiving antibiotics (n=29; group 1) and not receiving antibiotics (n=76; group 2). Articular fluid aspiration was performed under sterile conditions with an 18-gauge angiocath with at least 1 ml volume of the hip, knee, ankle, elbow, and shoulder joints. Polymorphonuclear cell percentage count, cell count, Gram staining (GS), culture, and leukocyte esterase test were performed immediately after the aspiration of the specimens. Levels of synovial fluid white blood cell count, serum white blood cell count, PMN, serum glucose, erythrocyte sedimentation rate, C-reactive protein, and time of aspiration (TOA) were significantly higher in the group receiving antibiotics (P<0.05). Synovial glucose levels were significantly lower in the group receiving antibiotics. Furthermore, the positive frequency of glucose esterase, blood culture, GS, serum culture, and ultimate diagnosis of septic arthritis tests were significantly lower in the antibiotic receiving group (P<0.05). The sensitivity, and positive predictive value of the leukocyte esterase test were obtained at 100%, and 96.55% in the antibiotic receiving group, respectively. Moreover, in the group not receiving antibiotics, the sensitivity, specificity, positive predictive value, and negative predictive value of the leukocyte esterase test were estimated at 72.22%, 92.50%, 89.66%, 78.72%, respectively. Antibiotic use and the prolongation of TOA lead to increased inflammatory products, which is interfering with lab variables. As a result, they increase the sensitivity of the test. The sensitivity and specificity of the leukocyte esterase test in patients who did not receive antibiotics showed that this was a suitable and reliable laboratory method for the rapid detection of infectious arthritis that required an emergency rescue procedure.

Effect of soil drenching with humic acid, L-methionine and phosphoric acid on Fusarium wilt and induction of enzymes related to oxidative stress and defense in tomato plants

The main objective of the present study was to evaluate the efficacy of humic acid, L-methionine and phosphoric acid as a soil application for controlling Fusarium wilt disease of tomato and their effect on gene expression alterations using protein and isozyme profiling. Two isolates from each of Fusarium oxysporum f. sp. lycopersici (FOL) and F. solani (FS) were obtained from diseased tomato plants. According to pathogenicity tests, all tested isolates were able to infect tomato cultivars; cv. Super Jackal and cv. 745 under greenhouse conditions. In in vitro studies, the addition of humic acid, L-methionine and phosphoric acid to the medium at different concentrations reduced the growth of the tested isolates of the pathogens. Under greenhouse conditions, the addition of humic acid at 2%, 2 g/L of L-methionine and 0.6% of phosphoric acid to infested soil after sowing significantly decreased percentage disease severity of Fusarium wilt at both seasons (2016/17 and 2017/18). Protein bands at molecular weight 47.2 KD, 38.2 KD, 16.3 KD and 15.6 KD appeared only in treated plants. New esterase, peroxidase, chitinase and superoxide dismutase isozyme bands were detected in all treatments while not appear in the control. This results revealed that these treatments were able to induced new protein which increases resistance in tomato plants to Fusarium wilt disease.

Fumigant toxicity of three Satureja species on tomato leafminers, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae)

The tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is one of the major insect pests of solanaceous crops especially tomatoes. As plants contain secondary metabolites with protective properties against unwanted agents, they can be used as insecticides after extraction from plants as essential oils (EOs). In this study, essential oils of three savory species, including Satureja khuzestanica, S. bachtiarica, and S. rechingeri were extracted. Analyzing the chemical compositions via gas chromatography–mass spectrometry (GC/MS) revealed that geraniol was the main component of all essential oils. The volatile activity of these three savory species on the fourth instar larvae of T. absoluta during 24 h, indicated that LC50 values of S. khuzestanica, S. bachtiarica, and S. rechingeri oils were 17.51, 25.03, and 34.33 μgL−1(air), respectively. Enzyme assays in vivo using alpha-naphthyl acetate (α-NA) as a substrate, showed that in the fourth instar larvae treated with 10 percent lethal concentration (LC10), general esterases of these larvae increased significantly in all treatments (p < .05). Polyacrylamide gel electrophoresis with a solution mixture of both substrates (α-and ß-NA) clarified three bands of esterases. However, this method failed to show any sublethal effects of the oils (LC10 value) on the larvae esterase bands. Considering the potential toxicity of these three species of Satureja essential oils to the tomato leafminers due to existence of several secondary metabolites, we can expect that new bio-pesticides can be developed from these promising plants in near future.

Morphological and biochemical responses to application of ascorbic acid on some lupine (Lupinus termis L.) cultivars under salinity stress

Salinity is abiotic stress and is an effective factor in the production of all crops, especially in semi-arid and arid areas. Ascorbic acid (ASA) is an important antioxidant in plant tissues and has a great role in tolerance to various stresses. The aim of this work is to study the effects of ascorbic acid on growth traits, leaf photosynthetic pigments, biochemical attributes such as total phenolic content, isozymes electrophoresis and protein electrophoresis of three lupine cultivars under salinity stress. The effects of ascorbic acid on germination percentage and different growth parameters, as well as on chlorophyll pigments, total phenolics, electrophoresis of protein and isozymes profiles of esterase (EST), peroxidase (POX) and catalase (CAT) in three cultivars of Egyptian lupine (Giza 1, Giza 2 and Giza 3), which were grown under salinity stress using concentrations of NaCl (0 and 50 mM), were investigated. The results showed a significant decrease in germination and all growth parameters, as well as chlorophyll pigments (Chl a, Chl b) and total phenolics (TP). Also, salinity stress showed noticeable changes in isozymes profiles and an increase in the total number of protein bands in all cultivars. Application of ascorbic acid (ASA) caused an increase in shoot length (SL), root length (RL), seedling fresh weight (SFW) and seedling dry weight (SDW) in all cultivars compared to the salinity without ASA. Also, pretreatment with ASA led to increase of Chl a, Chl b and TP as well as led to the induction of new isozymes bands of EST, POX and CAT that may be related to the tolerance of lupine cultivars to salinity stress. On the other hand, ASA stimulated the appearance of new protein bands and the disappearance of others with different molecular weights under salinity in all cultivars. It could be concluded that soaking lupine seeds in ascorbic acid at a concentration of 200 mg/L before being exposed to salinity reduced the adverse effects of salinity stress by improving morphological and biochemical characteristics.

Occurrence of Meloidogyne javanica as a Pathogen of Cockscomb (Celosia cristata) in Brazil

Cockscomb (Celosia cristata L., family Amaranthaceae) is an ornamental plant species cultivated in pots or used as cut flowers in floral arrangements. In April 2018, cockscomb ‘Comum’ exhibiting many galls on the roots was detected and sampled from several areas (27°21′34″S, 53°23′37″W) of Frederico Westphalen county, Rio Grande do Sul State, Brazil. Identification of Meloidogyne sp. (Nematoda: Meloidogynidae) was carried out by observations of perineal patterns (n = 30) and esterase phenotypes (n = 30), morphological measurements of second-stage juveniles (J2) (n = 50), and amplification and sequencing of the ITS1-5.8S-ITS2 rRNA region (primer set: forward 5′-TTGATTACGTCCCTGCCCTTT-3′ and reverse 5′-TCCTCCGCTAAATGATATG-3′) and the D2 to D3 fragment of the 28S rRNA gene (primer set: forward 5′-ACAAGTACCGTGAGGGAAAGTTG-3′ and reverse 5′-TCGGAAGGAACCAGCTACTA-3′). In addition, cockscomb roots were processed to determine the number of J2 and eggs of Meloidogyne sp. Nematode population density observed in the sample was 2,100 J2 per gram of cockscomb roots. The J2 had the following morphometric characteristics: length (L) = 405.5 ± 61.0 (368.5 to 415.6) µm, a = 29.6 ± 2.5 (26.8 to 33.4), c = 8.00 ± 0.5 (6.0 to 9.6), stylet = 13.5 ± 0.5 (11.3 to 15.9) µm, dorsal esophageal gland orifice (DGO) = 3.20 ± 0.4 (2.9 to 3.9) µm, tail length = 55.5.5 ± 4.0 (54.1 to 59.5) µm, and hyaline tail terminus = 10.3 ± 0.8 (9.0 to 11.5) µm. Female perineal patterns included a low trapezoidal dorsal arch and smooth striae interrupted by a pair of incisions on both sides. These lateral ridges that divide the dorsal and ventral striae are unique to Meloidogyne javanica (Treub, 1885) Chitwood, 1949 (Rammah and Hirschmann 1990). The polymorphisms in the esterase bands observed during electrophoresis revealed the typical phenotype of J3 (Rm = 1.00,1.25, 1.40) M. javanica populations (Carneiro et al. 2000). Sequence homologies were determined by comparing sequences of polymerase chain reaction (PCR) amplicons of D2 to D3 of 28S and ITS1-5.8S-ITS2 rRNA region that were noted to be 516 bp (GenBank MK072827) and 554 bp (MK072828) in length, respectively. The sequences of both markers exhibited 99 and 100% identity, respectively, with sequences being more similar to those of M. javanica isolates from Brazil, Vietnam, and China. The confirmation of nematode species identity was performed by PCR species-specific sequence characterized amplified region (SCAR) using a primer set composed of Fjav (5′-GGTGCGCGATTGAACTGAGC-3′) and Rjav (5′-CAGGCCCTTCAGTGGAACTATAC-3′). The PCR amplification using SCAR produced a specific fragment of expected size (∼670 bp) for M. javanica (Donkers-Venne et al. 2000). In greenhouse tests, cockscomb Comum plants being maintained in pots containing sterilized soil were inoculated with 5,000 eggs and J2 of the original M. javanica population; a noninoculated control was included, and treatments were replicated 10 times. After 60 days, all inoculated plants showed reduced growth compared with the control. Galling symptoms on the roots were similar to those observed in field-grown plants, and the nematode reproduction factor (final population/initial population) was 20.5. The noninoculated plants did not present gall formation in the roots, and their development was not affected either. The morphological and molecular characteristics of reisolated root-knot nematodes were identical to those of M. javanica. These results confirmed the nematode’s pathogenicity to cockscomb. To our knowledge, this is the first report on M. javanica parasitizing cockscomb in Brazil; therefore, our findings are of great importance to Brazilian flower growers to help them establish management measures M. javanica.

First Report of Meloidogyne graminicola (Nematoda: Meloidogynidae) on Barley (Hordeum vulgare) in Brazil

Winter barley (Hordeum vulgare L.) has become a growing addition to cereal crops in Brazil and, until recently, the root-knot nematode (Meloidogyne sp.) has not been reported to infect barley in this country. Meloidogyne graminicola is a major pathogen of rice (Oryza sativa L.) and causes significant yield losses in both upland and flooded fields in Rio Grande do Sul, Brazil (Negretti et al. 2017). In September 2017, many nematode galls were observed on the roots of barley plants (cv. BRS Brau), and samples were taken from areas in Santa Maria county (29°45′S; 53°38′W), Rio Grande do Sul State, Brazil. Meloidogyne sp. was identified using esterase phenotypes (n = 40), perineal patterns (n = 40), morphological measurements of second-stage juveniles (J2) (n = 50), and amplification and sequencing of the ITS1-5.8S-ITS2 rRNA region and the D2–D3 fragment of the 28S ribosomal RNA gene. The barley root systems were also processed to determine the number of J2s and eggs of Meloidogyne sp. The nematode population density observed in the samples was 2,585 eggs plus J2s per gram of barley root. The J2s had the following morphometric characteristics: length (L) = 458.2 ± 38.0 (390.0 to 476.0) µm, a = 26.6 ± 1.5 (24.9 to 29.4), c = 6.15 ± 0.4 (5.1 to 7.6), stylet = 14.2 ± 0.5 (13.4 to 15.0) µm, dorsal esophageal gland orifice (DGO) = 3.9 ± 0.4 (3.4 to 4.6) µm, tail length = 70.5 ± 4.7 (63.1 to 75.3) µm, and hyaline tail terminus = 19.5 ± 1.8 (14.5 to 25.5) µm. The female’s perineal patterns were oval with a low dorsal arch without a lateral line. The cuticular striations were smooth and thick in the dorsal region of the vulva, and the phasmids were close together (13.1 to 17.9 μm). These morphological characteristics matched the original description of M. graminicola (Golden and Birchfield 1968). The polymorphisms of esterase bands seen using electrophoresis revealed the phenotype VS-1 (Rm = 0.71) typical of M. graminicola. The sequence homologies were determined by comparing them with those from GenBank. PCR amplicons of D2–D3 of the 28S and ITS1-5.8S-ITS2 rRNA regions were 531 bp (GenBank MH707259) and 440 bp (MH703577), respectively. The sequences of both markers exhibited 99 and 100% identity, respectively, with sequences corresponding to M. graminicola isolates (D2–D3: MH843665 and KY660544; ITS: MH842692 and KY346887) from Brazil and China. Identification of the nematode species was confirmed by PCR using the species-specific sequence-characterized amplified region (SCAR) with primer sets GRAJ17-1F and GRAJ17-1R. PCR amplification using SCAR produced a specific fragment of the expected size (∼230 bp) in M. graminicola (Mattos et al. 2019). In greenhouse tests, barley plantlets from cv. BRS Brau that were maintained in pots with sterilized soil were inoculated with 5,000 eggs plus J2s of the original population of M. graminicola using 10 replicates; a noninoculated control was included in the test. After 60 days, the growth of all the inoculated plants was reduced compared with that of the control. Galling symptoms on the roots were similar to those in the field, with a nematode reproduction factor (final population/initial population) of 28.5. There were no galls on the roots of noninoculated plants, and plant development was not affected. The morphological and molecular characteristics of the reisolated root-knot nematode were identical to those of M. graminicola. M. graminicola has been reported on barley in India (Vaish et al. 2012). To our knowledge, this is the first report on M. graminicola parasitizing barley in Brazil. This finding is of great importance for Brazilian barley production because this nematode can severely damage barley plants and become a major problem for the cultivation of this crop.

Effect of cypermethrin on worker and soldier termites of subterranean termites Odontotermes brunneus (Hagen) (Termitidae: Isoptera)

The termite Odontotermes brunneus is an economically important species causing damage to cellulose containing wooden material and agricultural crops in India. Insecticide application is an effective strategy in termite control. In the present study the effect of cypermethrin was tested for workers and soldiers termite using filter paper dip method. After 24 h treatment the lethal concentration (LC50) was increased to 9.7 ppm in workers and 1.8 ppm in soldiers respectively. The detoxification enzyme activities of esterase, glutathione S-transferase was increased in worker termites 23 µmol, 9 µmol/min/mg of protein compared to soldiers 15 µmol, 7 µmol/min/mg of protein respectively (p < 0.05). The activity of mixed-function oxidase was found very less in both samples. Further nPAGE analysis revealed that increased esterase band in workers than soldier and control sample. The data of this study revealed that possible mechanism of esterase and glutathione S-transferase mediated cypermethrin detoxification that leads to reduce the sensitivity in worker termites of O. brunneus.

Identification of Morphological Character and Esterase Isozyme Pattern in Second-Generation Black Rice Plant Irradiated to Gamma Rays

Black rice is one of the functional foods due to its high anthocyanin content. Black rice grain was irradiated by gamma rays with a dose of 200 Gy and 300 Gy. The main purpose of this irradiation is to induce mutation to the black rice plant in order to achieve the improved organism. This study was undertaken to elucidate the morphological character and esterase isozyme pattern of black rice plant after irradiated by gamma rays. There were morphological differences on leaves, stems and grains between irradiated and non irradiated black rice plant. Gamma radiation dose of 200 Gy showed the significant influence of the length of the stem, number of internodes, and length of leaves. The radiation dose of 300 Gy showed the significant influence of the decrease value of diameter of 3rd internodes, number of branches and width of leaves. Flowering time is getting faster as increasing radiation dose. At the age of 74 days after planting there are 9.15% plants of 200 Gy radiation dose that have flowered faster than normal plants. This value increased into 11.45% at the dose of radiation 300 Gy. There were differences in the esterase banding pattern between radiation dose of 200 Gy and 300 Gy than the control plants, indicated that randomly mutation has occurred.

Electrophoretic Banding Pattern Of Esterase Isozyme In Different Tissues Of Puntius Sophore (Cyprinidae : Cypriniformes)

Esterase isozyme of different tissues of Puntius sophore was analyzed using 7.5 % polyacrylamide gel electrophoresis (PAGE). Fish specimens were collected from local market. The tissues used were taken from different muscles, stomach, fore-, mid- and hind-gut, liver, eyes, gill, heart, kidney, fore-, mid- and hind- brain, eggs and ovary. Six different esterase bands were detected, named Est-1, Est-2, Est-3, Est-4, Est-5 and Est-6 and their relative mobility were 1.0, 0.84, 0.62, 0.33, 0.26 and 0.13 respectively, each of them representing a single allele. The highest esterase activity was found in liver, followed by gill, kidney, heart, brain, intestine, stomach, eye, reproductive organ and skeletal muscles as detected in the staining intensity. Staining intensity of Est-4 and Est-5 was higher and Est-6 was the least stained in all the tissues.&#x0D; Asiat. Soc. Bangladesh, Sci. 42(2): 201-208, December 2016

Profiling Esterases in Mycobacterium tuberculosis Using Far-Red Fluorogenic Substrates.

Enzyme-activated, fluorogenic probes are powerful tools for studying bacterial pathogens, including Mycobacterium tuberculosis (Mtb). In prior work, we reported two 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one) (DDAO)-derived acetoxymethyl ether probes for esterase and lipase detection. Here, we report four-carbon (C4) and eight-carbon (C8) acyloxymethyl ether derivatives, which are longer-chain fluorogenic substrates. These new probes demonstrate greater stability and lipase reactivity than the two-carbon (C2) acetoxymethyl ether-masked substrates. We used these new C4 and C8 probes to profile esterases and lipases from Mtb. The C8-masked probes revealed a new esterase band in gel-resolved Mtb lysates that was not present in lysates from nonpathogenic M. bovis (bacillus Calmette-Guérin), a close genetic relative. We identified this Mtb-specific enzyme as the secreted esterase Culp1 (Rv1984c). Our C4- and C8-masked probes also produced distinct Mtb banding patterns in lysates from Mtb-infected macrophages, demonstrating the potential of these probes for detecting Mtb esterases that are active during infections.

Detection of carboxylesterase and esterase activity in culturable gut bacterial flora isolated from diamondback moth, Plutella xylostella (Linnaeus), from India and its possible role in indoxacarb degradation.

Diamondback moth (DBM), Plutella xylostella (Linnaeus), is a notorious pest of brassica crops worldwide and is resistant to all groups of insecticides. The insect system harbors diverse groups of microbiota, which in turn helps in enzymatic degradation of xenobiotic-like insecticides. The present study aimed to determine the diversity of gut microflora in DBM, quantify esterase activity and elucidate their possible role in degradation of indoxacarb. We screened 11 geographic populations of DBM in India and analyzed them for bacterial diversity. The culturable gut bacterial flora underwent molecular characterization with 16S rRNA. We obtained 25 bacterial isolates from larvae (n = 13) and adults (n = 12) of DBM. In larval gut isolates, gammaproteobacteria was the most abundant (76%), followed by bacilli (15.4%). Molecular characterization placed adult gut bacterial strains into three major classes based on abundance: gammaproteobacteria (66%), bacilli (16.7%) and flavobacteria (16.7%). Esterase activity from 19 gut bacterial isolates ranged from 0.072 to 2.32 μmol/min/mg protein. Esterase bands were observed in 15 bacterial strains and the banding pattern differed in Bacillus cereus – KC985225 and Pantoea agglomerans – KC985229. The bands were characterized as carboxylesterase with profenofos used as an inhibitor. Minimal media study showed that B. cereus degraded indoxacarb up to 20%, so it could use indoxacarb for metabolism and growth. Furthermore, esterase activity was greater with minimal media than control media: 1.87 versus 0.26 μmol/min/mg protein. Apart from the insect esterases, bacterial carboxylesterase may aid in the degradation of insecticides in DBM.

Determination of Fatty Acid Profiles and Esterase Activities in the Gills and Gonads of Vimba vimba (L., 1758)

AbstractThe main purpose of this study was to evaluate the gills and gonads of Vimba vimba in terms of fatty acid composition, esterase activity, and esterase band patterns by native‐PAGE method. In this context, C16:0 and C18:1n‐9 were the most abundant fatty acids in the TLs, NLs, and the PLs fractions of both tissues. EPA (C20:5n‐3) and DHA (C22:6n‐3) levels were higher in the gills than in the gonads. MUFAs were the main fatty acid groups in the TLs, NLs, and PLs of both tissues. Gills had higher n‐3/n‐6 ratios when compared with the gonads, with the exception of the PLs. TLs of the gonads had the highest AI and TI values. A concentration of 1 mM Na‐taurocholate increased the esterase activity of the gonads. Other chemicals having inhibition and surface active effects resulted in decreases in the esterase activities at concentrations of 1, 5, and 10 mM. In total, nine esterase bands were determined in both tissues. Four bands were shared by both the gills and gonads of V.vimba. These shared bands could be responsible for the similar esterase activities in both tissues. Gills and gonads of V. vimba have substantial amounts of fatty acids and esterase isozymes. More detailed studies are required to understand their industrial and nutritional uses.

Biochemical Characterization of Ridge Gourd and Sponge Gourd Genotypes

A genetic analysis of ridge gourd and sponge gourd genotypes were performed using protein profiling and isozymes analysis of four enzymes viz., peroxidase, polyphenol oxidase, esterase and super oxide dismutase. Fifteen day old seedlings of the genotypes were tested in Native-PAGE for the study of genetic diversity among them. Protein profiling of the genotypes developed nine bands having Rm values of 0.037, 0.075, 0.154, 0.213, 0.266, 0.326, 0.532, 0.584 and 0.663. Out of them, three bands were monomorphic and others were polymorphic. Seventeen ridge gourd and sponge gourd genotypes were grouped into two main clusters I and II with an average similarity of 47 per cent. Four bands of peroxidase isozymes were observed having Rm values of 0.083, 0.236, 0.515 and 0.803. Out of them two bands were polymorphic in nature. Four bands of polyphenol oxidase were detected with Rm values of 0.318, 0.438, 0.570 and 0.686. Out of them two bands were polymorphic in nature. Total five bands of esterase were observed having Rm values of 0.037, 0.102, 0.170, 0.269 and 0.372. Out of them four bands were polymorphic in nature and a band with Rm value 0.102 was monomorphic. Three bands of Rm values 0.246, 0.339 and 0.486 were observed for super oxide dismutase isozymes. The band of Rm value 0.486 was present in all the 17 genotypes and was monomorphic in nature.

Genetic diversity based on isozymic banding pattern in Heteropogon contortus - A perennial tropical forage grass

To ascertain the extent of diversity present among the different accessions of Heteropogon contortus-a perennial fodder grass, eighty-nine genotypes collected from different parts of India were compared for four enzymes, viz. peroxidase, superoxidase dismutase (SOD), glutamate oxalo-acetate transaminase (GOT) using starch gel electrophoresis and esterase using Polyacrylamide gel electrophoresis (PAGE). Biochemical markers such as isozymes are useful supplements in identifying the genetic variation in a crop. The study revealed presence of 15 peroxidase bands, 5 SOD bands, 9 esterase bands and 1 GOT band. The GOT band was monomorphic. Polymorphism was observed for all other bands indicating thereby existence of considerable genetic diversity. Frequency of bands varied widely among the genotypes. For peroxidase, band number 3 was found to be most frequent whereas, band number 2 and 14 were least frequent and observed in only 22 and 24 genotypes respectively. None of the bands were found to be species specific. For SOD, bands 1 and 5 were most frequent and observed in 80 and 83 genotypes respectively. While all the bands were represented in 11 genotypes, a single band was observed in 7 genotypes. For esterase, 9 polymorphic bands were distributed in two distinct migration zones. Zone 1 comprised of 9 bands with low mobility (rm value 0.41 to 0.49). Band no. 2, 6 and 7 were present in higher frequency, whereas bands 4 and 5 were present in only 7 and 8 genotypes respectively. For GOT, only one monomorphic band with rm value 0.65 was recorded. Clustering based on UPGMA analysis reveals that genetic diversity could not be correlated to the geographical origin of the accessions. The study indicates that considerable genetic diversity exists in the species and collection from different geographical places will help in identifying suitable species for various agro-ecological situations as suitable forage species for rangeland conditions.

Biodegradation of composite resin with ester linkages: Identifying human salivary enzyme activity with a potential role in the esterolytic process

ObjectivesThe ester linkages contained within dental resin monomers (such as Bisphenol A-glycidylmethacrylate (BisGMA) and triethylene glycol dimethacrylate (TEGDMA)) are susceptible to hydrolytic degradation by salivary esterases, however very little is known about the specific esterase activities implicated in this process. The objective of this work was to isolate and identify the dominant proteins from saliva that are associated with the esterase activities shown to be involved in the degradation of BisGMA. MethodsHuman whole saliva was collected and processed prior to separation in a HiPrep 16/60 Sephacryl S-200 HR column. The fraction with the highest esterase activity was further separated by an anion exchange column (Mono-Q (10/100G)). Isolated fractions were then separated by gel electrophoresis, and compared to a common bench marker esterase, cholesterol esterase (CE), and commercial albumin which has been reported to express esterase activity. Proteins suspected of containing esterase activity were analyzed by Mass Spectroscopy (MS). Commercially available proteins, similar to the salivary esterase proteins identified by MS, were used to replicate the enzymatic complexes and confirm their degradation activity with respect to BisGMA. ResultsMS data suggested that the enzyme fraction with the highest esterase activity was contained among a group of proteins consisting of albumin, Zn-α2-glycoprotein, α-amylase, TALDO1 protein, transferrin, lipocalin2, and prolactin-induced protein. Studies concluded that the main esterase bands on the gels in each fraction did not overlap with CE activity, and that albumin activity emerged as a lead candidate with significant esterase activity relative to BisGMA degradation, particularly when it formed a complex with Zn-α2-glycoprotein, under slightly basic conditions. SignificanceThese enzyme complexes can be used as a physiologically relevant formulation to test the biostability of composite resins.

English

Sesame is an orphan crop with little research attention in Kenya. Genetic relationship between cultivated sesame and related wild species in Kenya is not well known. The objective of this study was to determine genetic relationship between traditional landraces of sesame and related wild species using somatic chromosome counts and isozyme markers. Somatic chromosome counts of four wild species revealed a consistent chromosome number of 2n = 32, which differed from that of the cultivated sesame (2n = 26), indicating genetic variation in chromosome counts. Only esterase exhibited significant variation and accession-specific esterase bands were identified. Three cathodic and eight anodic bands were observed and the variable bands ranged from 2 to 6 per accession. Cathodic bands with varied relative migration were observed in wild species only, whereas anodic bands were observed for all the accessions. Accessions of cultivated sesame were more genetically diverse compared to wild species. Morogoro, 107UG, Indian-1 and Indian-2 recorded the highest number of esterase bands, while 103w had the lowest number of bands. Few common bands were found between cultivated sesame and related wild species indicating a distant genetic relationship. Few gene markers are available in sesame and related wild species, therefore, esterase isozymes can contribute to studies in the breeding and genetics of sesame. &nbsp; Key words: Genetic variation, isozyme, chromosome counts, wild species, sesame.

Changes of cotton leaf worm haemocytes and esterases after exposure to compounds derived from urea and Rice Straw

In the current study, the role of cotton leaf worm haemocytes and esterases in detoxification were recorded. The haemolymph was collected from the 6 th instar larvae which treated with LC25, LC50, LC70 of three newly compounds extracted from wastes from natural origin, Cyano acetyl urea (CAU), Benzimidazolyl acetyl urea (BAU) from urea and Cyano acetyl urea (CAH) from rice straw as 4 th instar larvae. The mean of total haemocytes counts in haemolymph of untreated 6 th larval instar was 9430±35.78 cells/mm 3 . Data indicated an increase in the total haemocytes of treated ones with CAU, at LC25 and LC50, BAU at LC25 and CAH at LC25, LC50 of CAH the percentages of change were: 19.01, 14.31, 1.90, 21.00 and 14.55, respectively. In addition, the percentages of oenocytoid counts were increased in all concentrations for all compounds except for CAH (at LC70) as it did not change comparing control count. On contrast, data cleared reductions in granulocytes in all concentrations for all compounds except for BAU at LC50 as they increased slightly (15.12%). On the other hand, the results were showed great differences in number of zones of esterase activity and in substrate specificity between treated and untreated samples. For instance, in the case of α - naphthyl acetate, 36 esterase bands were detected in control and treated larval samples with Rf ranging between 0.01 to 0.28. While, in the case of  - naphthyl acetate, 39 esterase bands were detected in control and treated larval samples with Rf ranging between 0.01 to 0.92.