Journal of BioScience and Biotechnology (JBB)

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https://doi.uni-plovdiv.bg/handle/store/2

p-ISSN: 1314-6238 / e-ISSN: 1314-6246

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3D structure prediction of lignolytic enzymes lignin peroxidase and manganese peroxidase based on homology modelling
(Plovdiv University Press “Paisii Hilendarski”, 2015-11-27) Kale, Swapnil K.; Deshmukh, Amit G.
Lignolytic enzymes have great biotechnological value in biopulping, biobleaching, and bioremediation. Manganese peroxidase (EC 1:11:1:13) and lignin peroxidase (EC 1:11:1:14) are extracellular and hem-containing peroxidases that catalyze H2O2-dependent oxidation of lignin. Because of their ability to catalyse oxidation of a wide range of organic compounds and even some inorganic compounds, they got tremendous industrial importance. In this study, 3D structure of lignin and manganese peroxidase has been predicted on the basis of homology modeling using Swiss PDB workspace. The physicochemical properties like molecular weight, isoelectric point, Grand average of hydropathy, instability and aliphatic index of the target enzymes were performed using Protparam. The predicted secondary structure of MnP has 18 helices and 6 strands, while LiP has 20 helices and 4 strands. Generated 3D structure was visualized in Pymol. The generated model for MnP and LiP has Z-score Qmean of 0.01 and -0.71, respectively. The predicted models were validated through Ramachandran Plot, which indicated that 96.1 and 95.5% of the residues are in most favored regions for MnP and LiP respectively. The quality of predicted models were assessed and confirmed by VERIFY 3D, PROCHECK and ERRAT. The modeled structure of MnP and LiP were submitted to the Protein Model Database.
A contribution to the knowledge of the trophic spectrum of three lacertid lizards from Bulgaria
(Plovdiv University Press “Paisii Hilendarski”, 2013-04-04) Mollov, Ivelin; Petrova, Slaveya
A study on the trophic spectrum of three species of lacertid lizards (Lacerta agilis, Lacerta trilineata and Podarcis muralis) was carried out, based on 20 specimens collected in the period 1967-1973 in various localities in Bulgaria. The analyzed data showed that the insects (Insecta) are the most numerous and the most frequently met among the alimentary components of the total amount of food of the studied stomachs (except for Lacerta agilis, where spiders are slightly predominating). The non-insect components consisted spiders and isopods. The largest niche breadth was recorded in Lacerta trilineata (8.25), followed by Podarcis muralis (5.20) and Lacerta agilis (3.44). The niche overlap between the three species (pair-wise comparison) showed medium values and in our opinion there should not be any serious competition for food resources at the places with sympatric distribution.
A contribution to the study of ground beetles (Coleoptera: Carabidae) in the Western Rhodope Mts. (Bulgaria)
(Plovdiv University Press “Paisii Hilendarski”, 2018-04-18) Teofilova, Teodora Marius
The available information on the diversity of the ground beetle fauna in the region of the Western Rhodope Mts. was enriched. A total of 133 species were found, including 16 endemic (12% of all) and 5 relict species. Ten species were new for the studied area: Amara morio nivium, Badister bullatus, Harpalus caspius, Laemostenus janthinus, Microlestes apterus, Pterostichus leonisi, Syntomus obscuroguttatus, Tachyura quadrisignata, Trechus asiaticus, Tachys micros. Genera Badister and Tachys were reported for the first time from the Western Rhodopes. Faunistic contribution was made for a region researched in detail for the last in 2006. The results proved the predominantly mesophilous nature of the habitats due to the large percentage of forest territories. The life forms of the carabids showed the prevalence of the zoophages over the mixophytophages both in qualitative (67%: 33%) and in quantitative terms (85.5%: 14.5%).
“A matter of health”: Evaluation of health habits in pupils in Primary School in Serbia
(Plovdiv University Press “Paisii Hilendarski”, 2018-03-14) Miličić, Dragana; Drndarski, Marina; Trajković, Jelena; Savić, Tatjana; Lučić, Luka; Pavković-Lučić, Sofija
We interviewed more than 400 primary school pupils from 5th to the 8th grade (aged 11 to 14 years) from the primary school in Belgrade (Serbia) about their habits concerning nutrition, beverage consumption, and time spend in front of a computer, tablet, playing games on consoles, or using a mobile phone. According to study, high percentage of students showed well-established habit of eating breakfast. Also, the overall water consumption prevailed in all grades, since the school ensures students access to safe and free drinking water, as a healthy alternative to consumption of sugar-sweetened beverages. However, we found increasing food consumption from bakery and consumption of carbonated drinks both in boys and girls from senior grades (7th and 8th). Also, they spent considerably more time with a computer, tablet, or mobile phone than the pupils from lower grades. About one-third of all surveyed students have been watching television or using a computer more than 3 hours during the day. We found significant increase of BMI, as an indicator of the increasing risk for weight gain and obesity both in boys and girls in senior grades. In the current curriculum of Biology, the topic of human health is studied in the 7th grade, but presented results show that one-year education in health topics is not sufficient. It suggests the necessity of introducing the topic of health in all grades of elementary school, starting from the lowest age.
A novel neutral protease from thermophilic Bacillus strain HUTBS62
(Plovdiv University Press “Paisii Hilendarski”, 2012-10-15) Aqel, Hazem; Al-Quadan, Farouk; Yousef, Tahani K.
A novel neutral highly thermostable protease was detected in the culture medium of thermophilic Bacillus strain HUTBS62 isolated from hot-spring located near to the Dead Sea, Jordan. The enzyme was purified by precipitation with 55-60% ammonium sulfate, gel filtration on Sephadex G-100 and DEAE ion exchange chromatography. The enzyme was purified 53-fold with 2% yield. The optimum pH and temperature for catalytic activity of protease was pH 6.8 and 80ºC, respectively, and 31% activity of protease remained even after heat treatment at 100ºC for 60 min. The relative activity of the enzyme was highly stable (90%) at 50ºC for 2 h. The half-life of the enzyme at 90ºC, 80ºC and 70ºC was estimated to be 3, 4 and 6 h, respectively. The activation energy of denaturation of purified enzyme was 21.7 kJmol-1. Iron, sodium, calcium, and manganese increased protease activity. On the other hand, magnesium, cobalt and zinc variably decreased the residual activity. But cadmium and copper drastically inhibited the enzyme activity. The enzymatic activity was highly stable in the presence of 1 and 2 mM EDTA at pH 6.8 and 80ºC. The neutral protease therefore could be defined as a highly thermostable with new properties make the present enzyme applicable for many biotechnological purposes.
A novel polymerase chain reaction (PCR) based assay for authentication of cell lines or tissues from human, pig and chicken origin
(Plovdiv University Press "Paisii Hilendarski", 2012-05-11) Gorenjak, Mario; Trapecar, Martin; Gradisnik, Lidija; Skok, Pavel; Cencic, Avrelija
A polymerase chain reaction based assay was developed for authentication of cell lines or tissues from human, pig and chicken origin. Specificity was achieved by species specific primer design targeting the mitochondrial D-loop sequence. Amplicon sizes were 114 bp, 169 bp and 645-648 bp for chicken, human and pig derived cell lines, respectively. Primers were tested for species specificity and non-specificity between haplogroups of the same organisms using BLAST tool and subsequently for cross amplification DNA extracted from human, chicken and pig venous blood as a positive control. Primers were also amplifying specific products in DNA extracted from individual cell line in both functional cell models and intentionally mixed cell lines consisting functional cell models. The PCR assay developed in this study represents a low-cost species specific end-point PCR based assay of the mitochondrial D-loop for the authentication of the cell line origin.
A preliminary study of the effect of phytoadditive carvacrol on the trace elements (Cu, Mn and Zn) content in fish tissues
(Plovdiv University Press “Paisii Hilendarski”, 2014-01-08) Yılmaz, Ebru; Yılmaz, Sevdan; Ergün, Sebahattin; Kaya, Hasan; Kızılkaya, Bayram; Soytaş, Nergiz
Phytoadditives have gained increasing interest as feed additives for fish. The aim of the present study was to determine whether selected dietary phytoadditive can influence the bioavailability of several trace elements (Cu, Mn and Zn), which play an important role in the physiological processes. The experiments were carried out at a commercial trout farm. A total of 420 juvenile rainbow trout (mean weight ± SD = 10.79±0.57), Oncorhynchus mykiss, were randomly allocated into four different treatments with three replicates each. Fish were kept in raceways (3X0.8X0.4 m) at 10±1°C with a natural photoperiod. Proper amount of carvacrol was sprayed on 1 kg of commercial trout diet to prepare four diets with 0 (Control, C0), 1 (C1), 3 (C3) and 5 (C5) carvacrol g/kg diet. Fish were fed to apparent satiation three times per day. The feeding trial lasted four weeks. Then, in different type of fish tissues (muscle, liver and pyloric caeca) from fish fed with diets enriched in carvacrol, beneficial elements (Cu, Mn and Zn) were analysed by atomic adsorption spectrophotometer. Results showed that the levels of Cu, Zn and Mn were especially significantly increase by C1 diet in all tissues (muscle, liver and pyloric caeca) except muscle and pyloric caeca Zn. The results of this experiment indicate that the carvacrol had the ability to potentiate the trace element retention. Although bioaccumulations of Cu, Zn and Mn in the muscle, liver and pyloric caeca are well demonstrated, the exact mechanisms of phytoadditives are still only partially understood. More investigations are required to detail the mechanisms involved in phytoadditives this enhancement.
A review of Sphaeropsis sapinea occurrence on Pinus species in Bulgaria
(Plovdiv University Press “Paisii Hilendarski”, 2017-07-11) Georgieva, Margarita; Hlebarska, Sianna
Sphaeropsis sapinea (Fr.) Dyko & Sutton causes shoot blight and canker disease throughout the world on conifers predisposed by stress. The disease is most important to Pinus species that are affected from the seedling stage in nurseries to mature trees in ornamental planting, forest plantations and natural stands. In Bulgaria, the first findings of the disease were noted in 1989 on Pinus nigra plantations in the North-eastern part of the country. Over the past few years, new emergency and severe damages have developed rapidly as a consequence of the prolonged drought periods during the last growing seasons. The high existence of S. sapinea outbreaks contributed considerably to the physiological weakness of pine trees that become more susceptible to attack by aggressive xylophages and other fungal pathogens. Disease occurrence and its pathogenicity are economically important affecting a number of pine trees from all ages. In Bulgaria, S. sapinea has been obtained on six pine species P. nigra, P. strobus, P. radiata, P. ponderosa, P. pinaster and P. halepensis. The aim of this review is to present the available knowledge on distribution, host specificity, biology, ecology, management of the disease, and to discuss its current prevalence and pathogenicity effect on pine species in Bulgaria.
Aberrant promoter methylation in p53 and ATM genes was not associated with sporadic breast carcinogenesis in Bulgarian patients
(Plovdiv University Press “Paisii Hilendarski”, 2014-04-02) Krasteva, Maria E.; Antov, Georgi G.; Gospodinova, Zlatina I.; Angelova, Svetla G.; Nacheva, Maria B.; Georgieva, Elena I.; Gavrilov, Ivan G.
Epigenetic silencing of gene expression has become an intensively studied topic regarding breast cancer during the last years. Previous investigations have shown that hypermethylation in gene promoters is an alternative to mutations cancer-related mechanism for inactivation of tumor suppressor gene function. The present study was designed for evaluation of promoter methylation in p53 and ATM tumor suppressor genes in Bulgarian patients with sporadic breast cancer. Sodium bisulfite conversion and methylation-specific PCR were performed on tumor DNA isolated from 55 patients with sporadic breast cancer. The data showed normal profile of methylation in the promoter region of p53 and ATM genes in all studied patients. Cases with hypermethylation were not observed. Our results did not support the involvement of p53 and ATM promoter hypermethylation in the neoplastic pathways of sporadic breast cancer. The lack of promoter hypermethylation in p53 and ATM is suggestive of a greater share of genetic aberrations in loss of tumor suppressor function of these genes and presumes the involvement of other epigenetic cancer mechanisms. To our knowledge this study provides the first data on promoter hypermethylation status of p53 and ATM genes in Bulgarian patients.
Aberrant signature patterns of ATM, γ-H2AX and p53 proteins in the patients exposed to methyl isocyanate diagnosed with gallbladder cancer
(Plovdiv University Press “Paisii Hilendarski”, 2014-04-29) Jatawa, Suresh Kumar; Tiwari, Archana
Cancer of gallbladder is a hidden phenomenon and highly malignant with underprivileged diagnosis and poor survival. Study of cancer patterns amongst victims of Bhopal gas tragedy exposed to methyl isocyanate revealed higher incidence of gallbladder cancer that necessitated a more objective elucidation of the disease at its molecular level. Tissues of 92 cases of gallbladder cancer patients were taken in the study (31 men and 61 women, age range 16–85 years, mean age 45.8±1.50 years). Mutations of ATM, γ-H2AX and p53 were predominantly seen in the methyl isocyanate exposed cohort diagnosed with adenocarcinoma, with 61.4% (43/70), 54.3% (38/70) and 73% (51/70) respectively, involving infiltration into the papillary and mucinous region/cell types of the gallbladder. Out of these, the expression frequency of all the above three genes was higher in moderately differentiated adenocarcinoma in comparison to poorly and well-differentiated ones. The results of the present study support the hypothesis that ATM and p53 mutations provide fundamental genetic signatures influencing tumor behavior across patient subsets and invasiveness of the disease, while γ-H2AX is apparently an ordinary pathway involved in the genesis of tumors.
Acetylcholinesterase inhibitory, antioxidant, and antimicrobial activities of Salvia tomentosa Mill. essential oil
(Plovdiv University Press “Paisii Hilendarski”, 2015-08-10) Marchev, Andrey; Ivanov, Ivan; Denev, Petko; Nikolova, Milena; Gochev, Velizar; Stoyanova, Albena; Pavlov, Atanas; Georgiev, Vasil
Chemical composition and bioactivity of essential oil from Salvia tomentosa Mill. natively grown in Bulgaria were investigated. GC-MS analysis identified 60 compounds which represented 98% of the oil constituents. The prevalent constituents were monoterpenes with eight dominant compounds being identified: borneol (10.3%), β-pinene (9%), camphor (7.9%), α-pinene (6%), camphene (4%), 1.8-cineole (3.8%), α-limonene (3.5%) and β-caryophyllene (3%). The essential oil showed considerable acetylcholinesterase inhibitory activity (IC50=0.28±0.06 μg/mL), comparable with that of galanthamine. Study of antioxidant activity strongly suggested that the hydrogen atom transfer reaction was preferable over the electron transfer (ORAC=175.0±0.40 μM Trolox equivalents/g oil and FRAP=1.45±0.21 mM Trolox equivalents/g oil). The essential oil showed moderate antifungal and antibacterial activities against Candida albicans and Gram-positive bacteria, whereas it was almost inactive against the investigated Gram-negative strains. The results suggested that the essential oil of Bulgarian S. tomentosa could be considered as a prospective active ingredient for prevention of oxidative stress-related and neurodegenerative disorders in aromatherapy. Because of the high antioxidant capacity, the oil could be considered as natural supplement or antioxidant in cosmetics and food products.
Adenovirus serotype 5 vectors with Tyr3-octreotate modified as a virulytic oncotherapy agent in AGS cells
(Plovdiv University Press “Paisii Hilendarski”, 2020-09-17) Salkhori, Najmeh Yarkeh; Farivar, Taghi Naserpour; Noroozi, Jamileh; Najafipour, Reza; Pakzad, Parviz
Background: Oncolytic viral therapy is known as a new promising strategy to treat cancer. Oncolytic viruses (OVs) can replicate in cancer cells, and beside this primary effect, OVs can also stimulate the immune system. Moreover, Adenovirus serotype 5 (Ad5) is widely used as an oncolytic agent for cancer therapy. The present study aimed to make a change in the adenovirus fiber area. Methods: The tyr3-octreotate sequence was inserted into the HI-loop in the adenovirus fiber knob. In addition, recombinant virus was transferred to the HEK-293 packaging cells using the calcium phosphate method; therefore, the new virus was prepared. Accordingly, virus titer was calculated using the TCID50 method. Afterward, recombinant adenovirus and wild-type 5 strain were transferred to perform against human adenocarcinoma gastric cell line (AGS). Subsequently, they were evaluated for cytopathic effects, and the manipulated virus was then confirmed Using PCR and sequencing. Results & Conclusions: In this regard, the obtained results showed that the recombinant adenovirus has a greater ability in infecting AGS cells compared to the wild-type adenovirus. Our data suggest that, modification of Ad5 with tyr3-octreotate expands its usage as an oncolytic agent against the AGS cells.
Agronomic performances and nutritional composition of three morphotypes of spider plant (Cleome gynandra L.) under different doses of compost
(Plovdiv University Press “Paisii Hilendarski”, 2019-07-15) Kiebre, Zakaria; Traore, Ernest Renan; Kiebre, Mariam; Bationo-Kando, Pauline; Kabore, Dramane; Sawadogo, Boureima; Sawadogo, Mahamadou
Spider plant is an important dietary supplement for the local population of Burkina Faso. The present study aims to evaluate the agronomic and biochemical response of three morphotypes to the variation of compost dose. The split-plot design with three repetitions was used for experiment. Three morphotypes, i.e slightly-violet, green and violet, were evaluated using four doses of compost. The results revealed that the agromorphological traits and biochemical composition vary considerably according to the compost dose. About 6t/ha was identified as the best for biomass production and nutritional profile of spider plant. In general, the agronomic response of morphotypes to the variation of compost dose is not significantly different, except biomass. However, under low fertility conditions, the green morphotype records the best performances whereas in high fertility conditions, the slightly-violet records the best performances. For the biochemical composition, the content of nutrients depends more of compost dose than morphotype.
Allelopathic tolerance in white lupine (Lupinus albus L.) accessions to Sorghum halepense extracts
(Plovdiv University Press “Paisii Hilendarski”, 2019-07-15) Georgieva, Natalia
Modern farming requires a better understanding of the response of species, and in the particular experiment, of accessions, to the influence of weeds in order to develop agricultural systems that are more dependent on ecological interactions and less dependent on the use of pesticides. The present lab study aimed to estimate the tolerance of white lupine accessions to Sorghum halepense extracts on the seed germination and initial growth of the crop. Studied factors were: white lupine accessions – ten levels (Lp01, Lp04, Lp06, Lp10, Lp21, Lp23, Lp25, Lp27, Lp28 Lp29), aqueous extracts – two levels (shoot and root biomass of S. halepense) and concentrations of the extracts – four levels (1.0%, 5.0%, 10.0% and distilled water as control). The results showed a different tolerance of the ten accessions to allelopathic action of the aqueous extracts. Increasing concentrations of the extracts inhibited seed germination (by 5.0 to 10% in the different accessions), elongation and accumulation of primary germ (2.4 to 71.2% and 3.0 to 56.2%, respectively. The GGEbiplot analysis, representing the summary effect of the action of weed extracts on the germination and growth parameters in different concentrations, determined Lp 01, Lp 04 and Lp 10 as tolerant. Sensitivity exhibited Lp 21, Lp 25 and Lp 23. The correlation analysis of data showed that accessions whose seeds had a higher 1000 seeds mass, higher seed vigor index and protein content were less affected by the inhibitory effect of S. halepense extracts, respectively, they exhibited a higher tolerance.
Allelopatic effects of some medicinal plant essential oils on plant seeds germination
(Plovdiv University Press “Paisii Hilendarski”, 2015-11-29) Shokouhian, Ali; Habibi, Hassan; Agahi, Kayvan
The effect of essential oils from some medicinal plants on seed germination was studied with the aim of assessing their potential use as bioherbicides. The experiment was conducted as factorial based on completely randomized design (CRD) with three replications. Seeds of 3 summer crops including lettuce (Lactuca sativa), pepper (Piper longum) and tomato (Solanum lycopersicum) were exposed to essential oils of rosemary (Rosmarinus officinalis), thyme (Thymus vulgaris) and anise (Pimpinella anisum) at 3 different concentrations (25 and 50% diluted and undiluted). Treated seeds were grown in a growth chamber at 25°C for 5 days. The number of germinated seeds in each Petri dish was daily counted. After five days seed germination percentage (Ge) was calculated. Biplot analysis was performed using genotype plus genotype environment interaction (GGE) method. Results showed that the allelopathic effect on Ge was varied among studied plants, which was mainly due to i) differences in the composition of the studied essential oils and ii) different allelopathic effects of the studied essential oils on Ge. Accordingly, compared to the individual use, combining several essential oils would have a greater inhibitory effect on Ge of weeds.
Allozyme genetic polymorphism in Bulgarian honey bee (Apis mellifera L.) populations from the south-eastern part of the Rhodopes
(Plovdiv University Press "Paisii Hilendarski", 2012-05-13) Ivanova, Evgeniya; Staykova, Teodora; Stoyanov, Ivan; Petrov, Plamen
Allozyme genetic polymorphism in Bulgarian honey bee populations from four different locations in the south-eastern part of the Rhodopes Mountain was studied on six enzymic systems (MDH, ME, EST, ALP, PGM and HK) corresponding to six genetic loci. Allozyme analysis revealed that all studied loci were polymorphic in almost all investigated populations. The observed heterozygosity was found to range from 0.110 to 0.208 and Nei’s genetic distance – between 0.016 and 0.061 among the studied populations. These honey bee populations were clustered in two groups in the UPGMA dendrogram. The Tihomir population was in a separate clade while other three populations (Kardzhali, Krumovgrad and Dolni Yurutci) were grouped together.
Alpha-glucosidase inhibitory effect and antioxidant properties of different extracts from Lycium barbarum L.
(Plovdiv University Press “Paisii Hilendarski”, 2019-02-14) Nikolova, Mariana; Alexandrov, Alexander; Iliev, Ilia
Fruits of the plant Lycium barbarum L. (Goji berry) fruit have been widely used around the world for their medicinal purposes and as a functional food for more than 4500 years. Studies indicate the effects of the fruits of Goji berry on aging, neuroprotection, fatigue, energy expenditure, glucose control in diabetics, antioxidant properties, immunomodulation, anti-tumor activity, and cytoprotection. Some authors report that antioxidant molecules found in goji fruit might offer many health protective beneﬁts by alleviating oxidative stress, when used as an additive or in a mixture with other phytochemicals in fruit or herbal products. It was found that Goji berry fruit growing in Bulgaria has a total polysaccharide content of about 4%. The content of total polyphenol compounds in the extracts was found to be between 5.5mg and 8mg GAE/g of dry product. Polyphenols from dry Goji berry fruit were extracted and studied by HPLC analysis for the type of polyphenols present. Antioxidant activity of extracts rich in polyphenols was determined by DPPH, FRAP and CUPRAC methods. The data was compared with a medicinal product containing an extract of Goji berry, where a synergistic reaction was also established. Inhibitory effects of the Goji berry extracts and their corresponding polyphenols were investigated in the presence of α-glucosidase for antidiabetic activity. The study demonstrated that some of the polyphenols in the investigated extracts exhibit competitive properties towards the enzyme.
Alternative substrates for higher mushrooms mycelia cultivation
(Plovdiv University Press “Paisii Hilendarski”, 2015-11-18) Krupodorova, Tetiana A.; Barshteyn, Victor Yu.
Cultivation of 29 species of higher mushroom mycelia on alternative substrates – wastes of Ukrainian oil-fat industry, has been investigated. The amount of mushroom mycelia obtaining on 12 investigated substrates varied significantly, from 1.0 g/L to 22.9 g/L on the 14th day of cultivation. The superficial cultivation adopted in this study allows for easy to choose appropriate medium (substrate) for mycelia production. Alternative substrates (compared to glucose-peptone-yeast medium) were selected for all studied species, from soybean cake – most suitable for the mycelial growth of 24 species, to walnut cake − suitable only for 2 species. The utilization of substrates has been evaluated by biological efficiency. The best index of biological efficiency varied from 19.0% to 41.6% depending on the mushroom species. It was established high biological efficiency of mycelia cultivation on substrates: wheat seed cake – Pleurotus djamor, Lyophyllum shimeji, Crinipellis schevczenkovi, Phellinus igniarius, Spongipellis litschaueri; oat seed cake – Ganoderma applanatum and G. lucidum; soybean cake – Hohenbuehelia myxotricha, Trametes versicolor, Morchella esculenta, Cordyceps sinensis, C. militaris, and Agrocybe aegerita; rape seed cake – Auriporia aurea; camelina seed cake – Fomes fomentarius. The cultivation of these species are perspective as a biotechnological process of agricultural wastes converted into mycelia, which could be used in different forms of products with therapeutic action: powder or tablets nutraceuticals or ingredients for functional foods.
An in silico analysis of the glutamate dehydrogenases of Teladorsagia circumcincta and Haemonchus contortus
(Plovdiv University Press “Paisii Hilendarski”, 2014-01-09) Brown, Simon; Muhamad, Noorzaid; Walker, Lisa R.; Pedley, Kevin C.; Simcock, David C.
Nematode glutamate dehydrogenase (GDH) amino acid sequences are very highly conserved (68-99% identity) and are also very similar to those of the bovine and human enzymes (54-60% identity). The residues involved in binding nucleotides or substrates are completely conserved and tend to be located in highly conserved regions of the sequence. Based on the strong homology between the bovine, Teladorsagia circumcincta and Haemonchus contortus GDH sequences, models of the structure of the T. circumcincta and H. contortus monomers were constructed. The structure of the T. circumcincta monomer obtained using SWISS-MODEL was very similar to that of the bovine enzyme monomer and the backbone of the polypetide deviated very little from that of the bovine enzyme monomer. Despite the sequence differences between the bovine and T. circumcincta enzymes, the relative positions and orientations of the residues involved in ligand binding were very similar. The reported Km for NADP+ of T. circumcincta is about 35 and times that of the bovine enzyme, whereas the Kms of the two enzymes for glutamate, -ketoglutarate and NAD(P)H are much more similar. The residue corresponding to S267 of the bovine enzyme is involved in binding the 2′-phosphate of NADP+ and is replaced in the T. circumcincta and H. contortus sequences by a tryptophan. The partial occlusion of the NAD(P)-binding site by the tryptophan sidechain and the loss of at least one potential H-bond provided by the serine may explain the lower affinity of the T. circumcincta for NADP+.
Analgesic effect of ethylacetate fraction of the methanol leaf extract of Hannoa klaineana in rats
(Plovdiv University Press “Paisii Hilendarski”, 2025-01-10) Abubakar, Ibrahim; Gusau, Mu’azu Abubakar; Danyaya, Jabir Aliyu; Ibrahim, Shehu; Abdullahi, Zayyanu; Zubairu, Abdulhakim; Sahabi, Abdulsalam Umar
Pain remains a major health, social, and economic problem worldwide. Hannoa klaineana Pierre & Engl. is a medicinal plant found in many African countries and used for the treatment of many diseases including pain-associated disorders. This study was conducted to evaluate the analgesic effect of the ethylacetate fraction of methanol leaf extract of Hannoa klaineana in rats. The analgesic effect of the ethylacetate fraction of methanol leaves extract of Hannoa klaineana (200, 400, and 600 mg/kg b.wt) was evaluated using acetic acid-induced writhing, tail flick (immersion), and hot plate model. In the acetic acid-induced writhing test, the extract (200, 400, and 600 mg/kg) demonstrated a significant (p < 0.05) decrease in the number of writhes with maximum percentage inhibition (75.61%) at 600 mg/kg dose of the extract. In tail flick and hot plate tests, the extract (200, 400, and 600 mg/kg b.wt) exhibited a significant (p < 0.05) increase in rats’ response with a steady increase in reaction time. Findings from this study show that ethylacetate fraction of the methanol leaves extract of Hannoa klaineana possessed analgesic activity which provided justification for the local use of the plant in the treatment of pain.

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