Major Polypeptides Research Articles

Relationships between protein and other chemical composition and texture of tofu made from soybeans grown in different locations.

Understanding quantitative relationships between protein and other chemical components in diverse soybean genotypes (lines) grown in different locations and the firmness of tofu can provide scientific insight for selecting soybean suitable for tofu making. Locations showed significant effects on seed components, including total protein, major storage proteins, subunits and polypeptides of the major storage proteins, and calcium, but not magnesium or phytic acid. Results showed that 11S content, but not 11S/7S ratio, was only correlated with filled tofu firmness when analyzed over all locations. A strong and positive correlation between firmness and A3 polypeptide of the 11S protein content was found for both pressed tofu (r=0.80, p<0.001) and filled tofu (r=0.76, p<0.001) over three locations (overall pooled data) and within most individual locations. The correlation of filled tofu firmness and A3 polypeptide was significant for each of the three individual locations. However, the correlation of pressed tofu firmness and A3 polypeptide content was significant at two of three locations. Mean calcium content was positively correlated with mean pressed and filled tofu firmness over all locations, but calcium was not correlated with pressed tofu firmness at any individual location, and only one location showed a significant correlation of calcium and filled tofu firmness. In addition, pressed tofu firmness was found to be negatively correlated with tofu yield. The findings that A3 polypeptide's strong relationship with tofu firmness within certain locations may be used by the food industry to select proper soybean for manufacturing tofu and to facilitate tofu soybean breeding for tofu making.

Isolation and Characterization of Submitochondrial Reticulum Complex Associated with Midpiece of Bovine Cauda Epididymal Sperm

The mitochondrial sheath of mammalian spermatozoa adheres to an underlying organized network of electron-dense material termed the submitochondrial reticulum (SMR). The objective of the present study is to characterize the structural components of the SMR. We have isolated SMR fractions from midpieces of bovine cauda sperm. SDS-PAGE of SMR fractions revealed four major polypeptides of 56kDa, 54kDa, 34kDa, and 31kDa. Proteomic analyses of the four polypeptides revealed that 56kDa and 54kDa polypeptides identified as glycerol kinase and 34kDa and 31kDa polypeptide identified as VDAC2 and VDAC3, respectively (Voltage-Dependent Anion-Selective Channel Protein 2/3). Western blots of midpiece stained with anti-56kDa, anti-54KDa, anti-34kDaVDAC2, and anti-31kDaVDAC3 antibodies demonstrated the presence of 56kDa, 54kDa, 34kDa, and 31kDa immunoreactive bands, respectively. Four polypeptides are localized to midpiece. Glycerol kinase activity was quantitively assayed in tails. We proposed that glycerol kinase and VDAC2/VDAC 3 polypeptides play an important role in sperm motility and in midpiece function.

Physicochemical and antigenic properties of the urea-extracted surface structures of &lt;i&gt;Yersinia pseudotuberculosis&lt;/i&gt; O:1b

Immuno-serological diagnostic tools particularly identifying pathogen antigens are the most important methods of pseudotuberculosis studies. The main immunodominant and species-specific antigens located in the surface structures of the bacterial cell are of practical interest. Thereby the aim of the work was to isolate and characterize biologically active surface structures of the pseudotuberculosis microbe. Here, the living cells of Y. pseudotuberculosis 3704 (O:1b) were lysed by using 9 M urea solution to extract antigens localized in the microbial surface structures. The subcellular fractions obtained such as outer membranes (OM), urea extract (UE) and isolated protein-lipopolysaccharide complex (PLPSC) are characterized by physical and chemical parameters. The protein content in the preparations ranged from 42% to 53%. The polypeptide band of the OM preparation, UE polypeptide and PLPSC for pseudotuberculosis microbe was presented by 14, 16 and 9 major polypeptides with molecular weight ranging from 13.9 kDa to 131.5 kDa, 13.5 kDa to 101.6 kDa, and 20.7 kDa to 66.6 kDa, respectively. Proteolytically active proteins and polypeptides were detected in isolated subcellular fractions (OM and UE) by using the radial enzyme diffusion test and substrate-gel electrophoresis found to be presented by 4 and 7 polypeptides with molecular weight ranging from 28.0 kDa to 118.0 kDa and 29.2 kDa to 97.7 kDa in the OM and UE preparation, respectively. The subcellular fractions obtained are capable to exhibit immunogenic activity after inoculation to experimental animals and antigenic activity while interacting with specific antibodies in the radial immunodiffusion (RID) assay and antibodies labeled with colloidal silver nanoparticles in dot immunoassay (DIA). OM and PLPSC preparations in DIA with immunoglobulins isolated from experimental antisera and labeled with colloidal silver nanoparticles were detected at a concentration of 0.12 g/ml (dry weight), cells of strain Y. pseudotuberculosis 3704 at a concentration of 3, 9 106 m.c./ml, which is similar to the results of DIA with immunoglobulins isolated from commercial pseudotuberculosis antiserum (St. Petersburg) and labeled with nanoparticles of colloidal silver. Thus, the subcellular fractions of pseudotuberculosis microbe isolated by using urea as a lysing and decontaminating agent retain their antigenic and immunogenic properties and enzymatic activity suggesting about their potential benefits for use to improve early diagnostics of pseudotuberculosis.

PRODUCTION OF ANTI-LACTOFERRIN ANTIBODIES AND THEIR APPLICATION IN ANALYSIS OF THE TEAR FLUID IN HEALTH AND CORNEAL INJURIES

Lactoferrin is a ubiquitous and multifunctional protein, which has antimicrobial and immunomodulatory activities. Lactoferrin plays an important role in the maintenance of ocular health. The aim of the study was to produce polyclonal antibodies against human lactoferrin in order to apply them in evaluation of lactoferrin levels in tear fluid collected from healthy eye and after corneal injury. Materials and methods. Affine chromatography on Protein A-sepharose was applied in order to isolate immunoglobulin G (IgG) fraction from the blood serum of lactoferrin-immunized rabbits. Each step of protein purification was monitored by denaturing gel electrophoresis (SDS-PAGE). Target antigen recognition by produced antibodies was established by western blot analysis with the use of diluted IgG fraction. Lactoferrin levels in the tear fluids collected from healthy individuals (n = 4) and patients with non-penetrating corneal injures (n = 6) were determined immunochemically with the use of purified antibodies. The results of western blot of lactoferrin levels in the tear fluids of healthy individuals and patients with corneal wounds were analysed using Mann-Whitney U-test. The difference between group mean values was considered significant at P&lt;0.05. Results. Using affine chromatography on Protein A-sepharose, antibodies against human lactoferrin were purified as IgG fraction from blood serum of lactoferrin-immunized rabbits. Western blot analysis showed that obtained antibodies recognize the antigen as a 75-kDa band, which corresponds to the intact human lactoferrin polypeptide. The same major polypeptide band was visualized by western blot with enhanced chemiluminescence detection in the tear fluid samples. Densitometry analysis of 75-kDa lactoferrin band showed 3.2-fold decrease in lactoferrin level in the tear fluid samples obtained from patients with non-penetrating corneal traumas as compared with samples collected from healthy persons (P&lt;0.05). Besides, tear fluid of patients with injured corneas contained large amounts of truncated lactoferrin immunoreactive polypeptides as well as high molecular weight bands, which could correspond to lactoferrin complexes with other proteins occurring during inflammation. Conclusions. According to our data, obtained anti-lactoferrin antibodies can be used as a valuable tool for development of advanced tests and procedures for diagnostics of eye diseases associated with the corneal lesions. Reduced lactoferrin concentration might represent a potential prognostic biomarker for diagnosis of ocular diseases including non-penetrating corneal injuries in a simple and non-invasive way.

Preparation and physicochemical properties of cucurbit protein isolate from Lagenaria siceraria defatted seed flour

Cucurbit seeds including Lagenaria siceraria seeds have been used from early times as a source of food. The protein constitutes about thirty-five percent of the weight of the decorticated seeds. Isolates are defined as the kind of protein that forms with a high concentration of protein that contains no dietary fiber. This study aims to investigate the preparation of protein isolated from Lagenaria siceraria defatted seed flour and to study their physicochemical properties. Protein was isolated from seed flours by using distilled water, salt solution, an alkaline solution, then precipitated with 0.1 N hydrochloric acid followed by centrifugation and freeze-drying. The findings reflected a high protein content (ranged from 80.67-91.04 g 100g-1) with the absence of fats. The functional properties of the protein isolate are said to be acceptable exhibiting a considerable range for all examined properties, water holding capacity 2.4-3.9 mL g-1 protein, fat binding ability 2.7-3.7 mL g-1 protein, emulsion capacity 51.0-57.8 mL of oil per g of protein, and bulk density 0.39-0.56 g mL-1, for all samples. The results of foaming stability indicated that the seeds were possibly utilized in food applications such as beverages and cakes processing. The maximum protein solubility (95%) occurred at the alkaline range pH 10 and11. The SDS page test detected that the three major polypeptides are those with molecular weights 10, 23, and 38 KDa. This study is the first to take place on the protein isolation from eight Lagenaria siceraria defatted seeds grown in Sudan, opening the way for further studies on these seeds

Immunoreactive polypeptide profiles of Sarcocystis fusiformis using hyperimmune and naturally infected serum

Immunoreactive polypeptide profiles study was made in the whole cyst lysate antigen of Sarcocystis fusiformis which was collected from buffaloes slaughtered at local slaughterhouse, Mathura, U.P. When whole cyst lysate antigen was subjected to SDS-PAGE, twelve major polypeptides (78, 66, 53, 50, 42, 39, 32, 29, 27, 24, 19 and 15kDa) were observed. On western blotting, out of twelve, six (32, 39, 42, 53, 66, 78kDa) and two polypeptides (53kDa and 66kDa) were found to be immunoreactive when probed with homologous hyperimmune and pooled known positive natural serum. No immunoreactivity was seen with pooled known negative natural serum. These two polypeptides (53kDa and 66kDa) bands can be explored for the development of sensitive and specific diagnostic assays.

Cloning, expression, solubilization, and purification of a functionally active recombinant cAMP-dependent protein kinase catalytic subunit-like protein PKAC1 from Trypanosoma equiperdum

The gene encoding the cAMP-dependent protein kinase (PKA) catalytic subunit-like protein PKAC1 from the Venezuelan TeAp-N/D1 strain of Trypanosoma equiperdum was cloned, and the recombinant TeqPKAC1 protein was overexpressed in bacteria. A major polypeptide with an apparent molecular mass of ∼38 kDa was detected by SDS-polyacrylamide gel electrophoresis, and immunoblotting using antibodies against the human PKA catalytic subunit α. Unfortunately, most of the expressed TeqPKAC1 was highly insoluble. Polypeptides of 36–38 kDa and 45–50 kDa were predominantly seen by immunoblotting in the bacterial particulate and cytosolic fractions, respectively. Since the incorporation of either 4% Triton X-100 or 3% sarkosyl or a mixture of 10 mM MgCl2 and 1 mM ATP (MgATP) improved the solubilization of TeqPKAC1, we used a combination of Triton X-100, sarkosyl and MgATP to solubilize the recombinant protein. TeqPKAC1 was purified by first reconstituting a hybrid holoenzyme between the recombinant protein and a mammalian poly-His-tagged PKA regulatory subunit that was immobilized on a Ni2+-chelating affinity resin, and then by eluting TeqPKAC1 using cAMP. TeqPKAC1 was functional given that it was capable of phosphorylating PKA catalytic subunit substrates, such as kemptide (LRRASLG), histone type II-AS, and the peptide SP20 (TTYADFIASGRTGRRNSIHD), and was inhibited by the peptide IP20 (TTYADFIASGRTGRRNAIHD), which contains the inhibitory motif of the PKA-specific heat-stable inhibitor PKI-α. Optimal enzymatic activity was obtained at 37 °C and pH 8.0–9.0; and the order of effectiveness of nucleotide triphosphates and divalent cations was ATP » GTP ≅ ITP and Mg2+ ≅ Mn2+ ≅ Fe2+ » Ca2+ ≅ Zn2, respectively.

Accessory Gene Products of Influenza A Virus.

Influenza A virus has long been known to encode 10 major polypeptides, produced, almost without exception, by every natural isolate of the virus. These polypeptides are expressed in readily detectable amounts during infection and are either fully essential or their loss severely attenuates virus replication. More recent work has shown that this core proteome is elaborated by expression of a suite of accessory gene products that tend to be expressed at lower levels through noncanonical transcriptional and/or translational events. Expression and activity of these accessory proteins varies between virus strains and is nonessential (sometimes inconsequential) for virus replication in cell culture, but in many cases has been shown to affect virulence and/or transmission in vivo. This review describes, when known, the expression mechanisms and functions of this influenza A virus accessory proteome and discusses its significance and evolution.

Striatin translocates to the cytosol of apoptotic cells and is proteolytically cleaved in a caspase 3-dependent manner

Striatin (STRN) is a multivalent protein holding great therapeutic potentials in view of its interaction with dynamic partners implicated in apoptosis. Although striatin-3 and striatin-4, that share high structural similarities with STRN, have been linked to apoptosis, the dynamics of STRN in apoptotic cells remain unclear. Herein, we report that the amount of STRN (110 kDa) is reduced in apoptotic cells, in response to various chemotherapeutic agents, thereby yielding a major polypeptide fragment at ~65 kDa, and three minor products at lower molecular weights. While STRN siRNA reduced the 65 kDa derivative fragment, the overexpression of a Myc-tagged STRN precipitated a novel fragment that was detected slightly higher than 65 kDa (due to the Myc-DDK tag on the cleaved fragment), confirming the cleavage of STRN during apoptosis. Interestingly, STRN cleavage was abrogated by the general caspase inhibitor Z-VAD.fmk. Cell fractionation revealed that the STRN pool, mainly distributed in the non-cytosolic fragment of naïve cells, translocates to the cytosol where it is proteolytically cleaved during apoptosis. Interestingly, the ectopic expression of caspase 3 in MCF-7 cells (deprived of caspase 3) induced STRN cleavage under apoptotic conditions. Inhibition of caspase 3 (Ac-DEVD-CHO) conferred a dose-dependent protection against the proteolytic cleavage of STRN. Collectively, our data provide cogent proofs that STRN translocates to the cytosol where it undergoes proteolytic cleavage in a caspase 3-dependent manner during apoptosis. Thus, this study projects the cleavage of STRN as a novel marker for apoptosis to serve pharmacological strategies targeting this particular form of cell death.

Small polypeptide hormones in plant

Generally the cell-to-cell communications take place by classical phytohormones such as auxin, cytokinin, ethylene, gibberellin, and abscisic acid. Likewise the plant peptide hormones are also involved in various aspects of growth and development. The physicochemical interactions of peptide hormones with their receptors activate downstream signaling which modulates cellular functions. These peptide actively involved in plant defense system improvements against biotic and abiotic stress (es), growth and development over the past decade have heralded the beginning of a new and potential growing field of polypeptide signaling in plants. Here, we review the currently known major plant polypeptides, their receptor proteins, polypeptide/receptor–mediated signaling cascades and their functions in plant.

Decellularized Tissue Matrix Enhances Self-Assembly of Islet Organoids from Pluripotent Stem Cell Differentiation.

Regenerating human islet organoids from stem cells remains a significant challenge because of our limited knowledge on cues essential for developing the endocrine organoids in vitro. In this study, we discovered that a natural material prepared from a decellularized rat pancreatic extracellular matrix (dpECM) induces the self-assembly of human islet organoids during induced pluripotent stem cell (iPSC) pancreatic differentiation. For the first time, we demonstrated that the iPSC-derived islet organoids formed in the presence of the dpECM are capable of glucose-responsive secretion of both insulin and glucagon, two major hormones that maintain blood glucose homeostasis. The characterization of the organoids revealed that the organoids consisted of all major endocrine cell types, including α, β, δ, and pancreatic polypeptide cells, that were assembled into a tissue architecture similar to that of human islets. The exposure of iPSCs to the dpECM during differentiation resulted in considerably elevated expression of key pancreatic transcription factors such as PDX-1, MAFA, and NKX6.1 and the production of all major hormones, including insulin, glucagon, somatostatin, and pancreatic polypeptide from stem cell-derived organoids. This study highlights the importance of natural, bioactive biomaterials for building microenvironments crucial to regenerating islet organoids from stem cells.

Composition and some functional properties of Bambara groundnuts vicilin fraction

Bambara groundnut is a drought tolerant pulse grain and an alternative source of protein. In this study, Bambara vicilin (7S) protein fraction was extracted and characterised in comparison to the storage protein. The proline content of vicilin fraction (4.3 g/100 g protein) was almost four times that of the storage protein. Fluorescence and hydrophobicity data suggested a less folded structure for the vicilin fraction when compared to the storage protein. Gel electrophoresis indicated three major polypeptides (50, 70 and 80 kDa) in the vicilin fraction with the presence of disulfide bond(s) in one of the strands. In contrast to the storage protein, which had <40% protein solubility at pH 2–9, vicilin had a significantly (p < 0.05) higher solubility with a minimum of 50% at pH 5 and maximum of 82% at pH 3. Emulsifying stability of the proteins significantly differed (p < 0.05) at pH 7. Foaming capacity of the vicilin fraction was significantly (p < 0.05) higher than that of the storage protein at pH 3, 7, and 9. It could be concluded that Bambara vicilin fraction could serve as a potential ingredient in the formulation of food products at acidic pH.

Proteomic analysis of decellularized pancreatic matrix identifies collagen V as a critical regulator for islet organogenesis from human pluripotent stem cells

In pancreatic tissue engineering, generating human pancreatic islet organoids from stem cells has been challenging due mainly to a poor understanding of niches required for multicellular tissue self-assembly in vitro. In this study, we aimed to identify bioactive, chemically defined niches from natural, biological materials for islet development in vitro. We investigated the proteomics of decellularized rat pancreatic extracellular matrix (dpECM) hydrogel using advanced bioinformatics analysis, and identified that type V collagen (ColV) is constantly and abundantly present in dpECM hydrogel. Niches provided to human pluripotent stem cells (iPSCs) by presenting ColV in matrix coating substrates permitted stem cells progression into islet-like organoids that consist of all major pancreatic endocrine cell types, i.e. α, β, δ, and pancreatic polypeptide cells. In the presence of ColV niches, gene expressions of all key pancreatic transcription factors and major hormone genes significantly increased in iPSC-derived organoids. Most importantly, ColV-containing microenvironment resulted in enhanced glucose responsive secretions of both insulin and glucagon hormone from organoids. The study demonstrates that ColV is a critical regulator that augments islet self-assembly from iPSCs, and it is feasible to utilize natural biomaterials to build tissue cues essential for multicellular tissue production in vitro.

Effect of steaming and dehydration on the nutritional quality and functional properties of protein isolates produced from Lablab purpureus (L.) Sweet (hyacinth bean)

The study conducted was focused on investigating the effect of steaming and dehydration on the nutritional quality and functional properties of protein isolates produced from hyacinth bean. This was for obtaining value-added legume-derived protein ingredients for potential food applications. Protein isolates prepared by isoelectric precipitation from legume grains subjected to steaming and dehydration prior to soaking, respectively, displayed traits of high-quality proteins. The major protein polypeptides present among samples were vicilin (7S) and legumin (11S). Steaming was identified useful for producing protein ingredients for nutritional intervention with potential health benefits. Samples recorded improved Fischer's ratio (1.57–1.78), BCAA to tyrosine ratio (3.51–4.28), predicted biological values (44.84–54.70), foaming, and emulsion properties. Dehydration extensively preserved the nutritional qualities and functionality of steamed samples. Findings have the potential to diversify the use of hyacinth bean and help improve the competitiveness of the legume grain sector. Practical applications This study presents how processing strategies (soaking, steaming, and dehydration) may be employed for obtaining legume-derived protein ingredients with improved nutritional quality and functional properties for potential food applications. The underutilization of many indigenous crops is mainly attributed to both limited research and marketing. Thus, research findings could potentially improve the value of hyacinth bean by diversifying its use and help improve the competitiveness of the legume grain sector. This could be achieved through demand for possible food applications that require protein enrichment. Furthermore, results may assist in broadening dehydration technology preferences in existing markets for obtaining legume grain-based ingredients or products with maximum quality.

Molecular characterization of yolk proteins in the female crab Neptunus pelagicus (A. Milne-Edwards, 1861) from the Mediterranean Sea of Alexandria, Egypt

This study aimed to characterize the morphological changes in the ovary of the female crab Neptunus pelagicus and to identify specific fractions of vitelloginin and vitelline molecules during primary and secondary vitellogenesis. Samples of the blue crab were collected from the Mediterranean Sea of Alexandria monthly during 2017. Ovaries and oocytes in primary and secondary vitellogenesis were detached and treated for histological test. Native polyacrylamide gel electrophoresis (PAGE) Bis-Tris Gels was applied to identify vitelloginin (VN) and vitelline (VL) molecules. Protein Analyses were done by PAGE-SDS. The initial degenerate primers were built regarding the conserved amino acid domains of the yolk proteins. Primary and secondary vitellogeneses consisted of 8 phases. Lipoprotein fraction with molecular weight 550 kDa was identified in the hemolymph in secondary vitellogenesis. Two protein fractions (VLI & VLII) were identified in secondary vitellogenic oocytes. The electrophoresis performed with extract of stage I oocyte showed two protein fractions with molecular weights 550 kDa and 460 kDa. In stage II and III oocyte, 4 subunits were presented of 180, 195, 140 and 120 kDa in VLI and 2 subunits with molecular weight of 110 kDa and 95 kDa in VLII. Another two fractions in stage V oocyte presented with molecular weights of 380 kDa and 360 kDa. Western blot analysis proved that both fractions were of four major polypeptide subunits with molecular weight of 180, 125, 90 and 85 kDa in each of the two VLs. The hybridization signal obtained by the Northern blot was detected in the hepatopancreas during ovarian cycle and in the ovary during secondary vitellogenesis. The result of the reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that the mRNA that encodes the C-terminal region of the VN cDNA was found in the ovary in secondary vitellogenesis and in the hepatopancreas.

Isolation and partial structural characterization of new Kunitz-type trypsin inhibitors from the pike cestode Triaenophorus nodulosus

The inhibitors produced by the parasitic worms successfully protect them from the host’s proteases and are supposed to underlie the host-parasite specificity. Our previous study has shown that the extracts from the pike tapeworm Triaenophorus nodulosus inhibit host proteinases and commercial trypsin. We aimed to isolate and identify the components responsible for trypsin inactivation. After a two-step separation the molecular masses were measured by SE-HPLC. The sample proved to contain four fractions represented by polypeptides (1–45 kDa) and low-molecular hydrophobic compounds. According to SDS-PAGE analysis, the major polypeptides in the fractions displaying the highest inhibition had masses of 14.4 kDa. The study culminated in partial N-terminal amino acid sequence analysis with a further search for homology. The research revealed two novel Kunitz-type proteins potentially responsible for the inhibitory capacity of the tapeworms against trypsin. Our findings extend the list of cestodes relying on Kunitz-type proteins in the host-parasite molecular cross-talk.

Effect of high-speed shearing treatment on dehulled walnut proteins

To understand more clearly about true characteristics of walnut protein, dehulled walnut kernels were used to prepare proteins, thus influences of polyphenols on proteins were minimized. Based on solubility, glutelin was found to be the predominant walnut protein (67.64% of total walnut proteins), meanwhile albumin and globulin accounted for 32.29%. SDS-PAGE and two-dimensional electrophoresis results showed that polypeptide profiles for albumin, globulin and glutelin exhibited significant differences. Walnut protein contained major polypeptides of MWs 30–35 kDa (pI 5.5–7.5) and 17–22 kDa (pI 9.0–10.0), along with minor polypeptides of MWs 50–73 kDa (pI 6.0–7.5) and 11–13 kDa (pI about 6.3). Disulfide bonds-linked polypeptides in walnut protein were determined with diagonal electrophoresis, and intermolecular disulfide bonds among polypeptides with MWs 30–34 kDa and 17–22 kDa were dominant. Further analysis by MALDI-TOF/TOF-MS showed that main components of walnut proteins were vicilin and legumin-like proteins. Through high-speed shearing (15,000 rpm) treatment at 60 °C for 5 min, walnut protein solubility were greatly increased from 24.33% to 62.81% at pH 8.0 and from 50.93% to 76.65% at pH 9.0. High-speed shearing treatment, especially at basic pH, could be a better way to improve protein solubility, with decreased particle sizes and more globulin and glutelin solubilized.

WDP formulations using a novel mosquitocidal bacteria, Bacillus thuringiensis subsp. israelensis/tochigiensis (VCRC B-474) – Development and storage stability

A novel mosquito active strain, Bacillus thuringiensis (VCRC B474) sharing the antigens of 2 serotypes, namely israelensis &tochigiensis was characterized by scanning electron microscopy and SDS-PAGE. The spherical and ovoid crystals present in this strain was composed of major polypeptides the size of 28, 65, and 130 kDa respectively. The sporulated cell mass was formulated into water dispersible powder (WDP) formulations with different carrier materials and checked for activity against Culex quinquefasciatus larvae at monthly intervals for up to a year. The formulation containing chalk was the most effective with LC50 values ranging between 0.274–0.523 μg/ml compared to the formulations containing bentonite (0.335-0.775) μg/ml and talc (0.348-0.808 μg/ml). The decline in the activity of these formulations with storage period was as follows: 3 months −14%, 22%, 20% respectively, 6 months - 25%, 35%, 37% respectively, 9 months − 39%, 50%, 47% respectively and 12 months −52%, 43%, 40% respectively. This study demonstrated that wet biomass of bacterial isolates could be simply mixed with carrier materials, dried and used for mosquito larval control without significant loss of activity for up to 6 months at room temperature. Further, this strain of Bacillus thuringiensis var. israelensis/tochigiensis (H14/19) can be a prospective candidate for use in mosquito control programs.

PUBLIC HEALTH HAZARDS ASSOCIATED WITH NATURALLY OCCURRING TOXINS IN LEGUME SEEDS - RAPID DETECTION AND CHARACTERIZATION OF A LECTIN FROM A KOREAN CULTIVATED KIDNEY BEAN (PHASEOLUS VULGARIS L.)

Objective: The delicacy of kidney beans is highly appreciable but, at the same time, their toxicity has raised an alarming concern. In this regard, more public awareness is needed about bean poisoning.Methods: This work aimed at promptly investigating the chemical and biological properties of a lectin derived from a Korean cultivar of the common bean (Phaseolus vulgaris L.).Results: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of two major polypeptide bands around 31.00 kDa. Hemagglutination assay and Ouchterlony double immunodiffusion technique confirmed the presence of lectin in dry seeds.Conclusion: It was evident that the Korean bean cultivar contains biologically active lectins, which may pose a risk to the consumer if the beans are eaten in raw or undercooked state.

Immune state correlates with histopathological level and reveals molluscan health in populations of Modiolus kurilensis by integral health index (IHI).

Quantitative analysis of the histopathological and immune parameters of bivalve Modiolus kurilensis collected from water areas with different level of ecotoxicological stress was performed. Significant differences between samples from polluted and non-polluted sites were revealed for total haemocyte count; percentage of agranulocytes; size and internal complexity of agranulocytes and granulocytes; phagocytic activity; percentage of NBT-positive cells; hemolytic activity and plasma protein concentration; percentage of the optical density of haemolymph major polypeptide bands at 55 kDa, 78 kDa, and 124 kDa; concretion coverage area in the kidney tubules; thickness of the tubular basement membrane; nephrocyte shape; and karyopyknosis of the kidneys; and hypervacuolisation; necrosis; karyopyknosis; haemocyte infiltration; fibrosis; and invasion of the digestive gland. Analysis of the global histopathological condition index based on the weighted indices also revealed that both the digestive gland and kidneys showed significantly greater histopathological changes in the bivalves collected from polluted water. Bivalve histopathology is an established tool in aquatic toxicology. However, it reflects a morphological picture of change, which, as a rule, can be clearly recorded only at the later stages of pathology, and in some cases, indicates an adaptation to stressors within the physiological norm. In this respect, a promising and highly sensitive biomarker of the functional state of bivalves, in terms of norm and pathology as well as their habitat, is the evaluation of immune status in combination with morphological changes. However, the use of different methods and scales of assessment and the diagnosis of biomarkers, characterised by different profiles of the stress response, makes it difficult to compare the results of different studies. We propose a reliable and powerful system for assessing the physiological state of bivalve molluscs, expressed in the integral health index (IHI) and based on the standardisation of the numerical values for all parameters that have significant differences between animals collected from impacted and non-impacted water areas. In our study, IHI calculated in three variants (for histopathological parameters, for immunological parameters, and in combination) showed the most significant differences in each of the cases, but the strongest difference (-4.07) was in calculating the total IHI, which included both the immune and histopathological parameters (p = 0.00005).