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Protein–drug interactions are crucial for understanding drug delivery and cell functions. Jacalin is a suitable molecule for such targeting, as it specifically recognizes the tumor-associated Thomsen–Friedenreich (TF) antigen that is expressed on the glycosylated proteins in cancer cells. The present paper describes the interaction of curcumin and jacalin, a possible carrier molecule for the delivery of antitumor drugs due to its ability to recognize tumor cells. Our results have shown that both steady-state fluorescence and fluorescent labelling of jacalin are two reliable methods to determine jacalin-curcumin interactions. The affinity of jacalin for curcumin is consistently within the micromolar range (using fluorescence and microscale thermophoresis) showing high-affinity binding of the complex. In vitro experiments on triple-negative breast cancer MDA-MB-231 cells indicated inhibition of cell growth after treating with the jacalin-curcumin complex for 48 h. The cell survival fraction was significantly reduced to 50% after combined treatment. In this paper, we report for the first time about the jacalin-curcumin interaction. We quantified this unique biomolecular interaction and gathered additional information on the binding event. We observed that the jacalin-curcumin complex inhibits the proliferation of the triple-negative breast cancer MDA-MB-231 cells. [ABSTRACT FROM AUTHOR]

High salinity is a major stress factor affecting the quality and productivity of rice (Oryza sativa L.). Although numerous salt tolerance-related genes have been identified in rice, their molecular mechanisms remain unknown. Here, we report that OsJRL40, a jacalin-related lectin gene, confers remarkable salt tolerance in rice. The loss of function of OsJRL40 increased sensitivity to salt stress in rice, whereas its overexpression enhanced salt tolerance at the seedling stage and during reproductive growth. β-glucuronidase (GUS) reporter assays indicated that OsJRL40 is expressed to higher levels in roots and internodes than in other tissues, and subcellular localization analysis revealed that the OsJRL40 protein localizes to the cytoplasm. Further molecular analyses showed that OsJRL40 enhances antioxidant enzyme activities and regulates Na+-K+ homeostasis under salt stress. RNA-seq analysis revealed that OsJRL40 regulates salt tolerance in rice by controlling the expression of genes encoding Na+/K+ transporters, salt-responsive transcription factors, and other salt response-related proteins. Overall, this study provides a scientific basis for an in-depth investigation of the salt tolerance mechanism in rice and could guide the breeding of salt-tolerant rice cultivars. [ABSTRACT FROM AUTHOR]

Proteins that carry specific carbohydrate-binding lectin domains have a great variety and are ubiquitous across the plant kingdom. In turn, the plant cell wall has a complex carbohydrate composition, which is subjected to constant changes in the course of plant development. In this regard, proteins with lectin domains are of great interest in the context of studying their contribution to the tuning and monitoring of the cell wall during its modifications in the course of plant organ development. We performed a genome-wide screening of lectin motifs in the Zea mays genome and analyzed the transcriptomic data from five zones of primary maize root with cells at different development stages. This allowed us to obtain 306 gene sequences encoding putative lectins and to relate their expressions to the stages of root cell development and peculiarities of cell wall metabolism. Among the lectins whose expression was high and differentially regulated in growing maize root were the members of the EUL, dirigent–jacalin, malectin, malectin-like, GNA and Nictaba families, many of which are predicted as cell wall proteins or lectin receptor-like kinases that have direct access to the cell wall. Thus, a set of molecular players was identified with high potential to play important roles in the early stages of root morphogenesis. [ABSTRACT FROM AUTHOR]

Human chorionic gonadotropin (hCG) is a dimeric, highly glycosylated hormone with a total of 4 N- and 4 O-glycosylation sites in its two subunits, hCGα and hCGβ. Recently, we developed a novel nano liquid chromatography coupled to high resolution mass spectrometry (nanoLC-HRMS) method for the analysis and thus the detection of the intact glycoforms of hCG. Here, a sorbent functionalized with the Jacalin lectin was evaluated in solid-phase extraction (SPE) for its potential to fractionate the hCG glycoforms prior to their nanoLC-HRMS analysis at the intact level, which may facilitate the detection of low-abundance glycoforms and may lead to a more detailed characterization of the hormone glycosylation. A commercial sorbent based on Jacalin immobilized on Sepharose and having a lectin density of 4.5 mg per ml of gel was selected to carry out SPE and its capacity was estimated to be of some tens of μg of hCG per ml of lectin sorbent. Next, the SPE protocol was modified to improve the extraction recoveries. Especially, it was noticed that an extensive pre-conditioning procedure prior to the first use of a cartridge was necessary to remove the residual non-grafted lectins. Indeed, if non-grafted lectins are not eliminated, they may bind a part of hCG glycoforms preventing their retention by the sorbent, leading to low extraction recoveries (around 10 %). With the extensive pre-conditioning procedure, the average extraction recoveries for both hCGα and hCGβ glycoforms were about 50 %, with either recombinant or urinary hCG. Qualitatively, the fractionation of hCG glycoforms between the washing and elution fractions was achieved with the urinary hCG sample by determining the number of glycoforms detected in each fraction. It appears that 12 hCGα glycoforms have a low affinity (detected only in the washing fraction), 1 a low-medium affinity (detected in washing and elution 1 fractions), 16 a medium affinity (detected in washing, elution 1 and 2 fractions), and 12 a high affinity (detected only in elution 1 and 2 fractions). For the hCGβ glycoforms, similarly, 3 have a low affinity and 12 a low-medium affinity. Additionally, the 3 hCGβ glycoforms were detected better. A different behavior was observed with the recombinant hCG sample, which indicates glycosylation differences between the two hCG samples. This shows the potential of lectin-based affinity fractionation before nanoLC-HRMS analysis to better characterize the glycosylation state of hCG at the intact level. • Capacity of Jacalin sorbents were evaluated with recombinant and urinary hCG. • the SPE procedure was developed to improve extraction recoveries. • SPE with Jacalin sorbents led to recoveries of around 50 % for hCG glycoforms. • Jacalin SPE enabled fractionation of hCG glycoforms as a function of their affinity. • lectin SPE and intact protein analysis by nanoLC-HRMS improves hCG characterization. [ABSTRACT FROM AUTHOR]

Aims: This study aimed to evaluate the combined effect of a mannose‐binding lectin Helja with fluconazole (FLC) on Candida albicans and to get insights about the joint action mechanism. Methods and Results: The fungal growth was assessed following the optical density at 630 nm. Fungal cell morphology and nucleus integrity were analysed by flow cytometry and confocal laser scanning microscopy using Calcofluor White (CFW) and 4′,6‐diamidino‐2‐phenylindole (DAPI) staining respectively. The basis of Helja + FLC action on cell wall and plasma membrane was analysed using perturbing agents. The Helja + FLC combination exhibited an inhibitory effect of fungal growth about three times greater than the sum of both compounds separately and inhibited fungal morphological plasticity, an important virulence attribute associated with drug resistance. Cells treated with Helja + FLC showed morphological changes, nucleus disintegration and formation of multimera structures, leading to cell collapse. Conclusions: Our findings indicate that the Helja + FLC combination exhibited a potent antifungal activity based on their simultaneous action on different microbial cell targets. Significance and Impact of Study: The combination of a natural protein with conventional drugs might be helpful for the design of effective therapeutic strategies against Candida, contributing to minimize the development of drug resistance and host cell toxicity. [ABSTRACT FROM AUTHOR]

Introduction: Lectins are hormone consists of carbohydrate binding proteins that perceive explicit epitopes present on certain glycoproteins and play roles in metabolism. Materials and Methods: Here we examined the localization of two different lectins, ghrelin and jacalin, in rat kidney of both sexes at neonatal 20 days and postnatal 1, 2, 4, 6, 8 and 12 weeks by immunohistochemistry. Results: Ghrelin and jacalin were found in the intrarenal kidney from neonatal day 20. Ghrelin immunoreactivity was mild in the proximal convoluted tubule (PCT), slightly stronger in the distal convoluted tubule (DCT), and absent in the renal corpuscle (RC). In contrast, jacalin immunoreactivity was intense in the RC but absent in the PCT and DCT. At postnatal one week, the ghrelin intensity increased and started to appear in the glomerular capillary of the RC and collecting duct (CD), while the jacalin immunoreactivity was intense in the RC, collecting tubules (CT), and loop of Henle. At postnatal two weeks, ghrelin immunoreactivity was intense in the DCT, CT, and loop of Henle, while jacalin immunoreactivity was intense in the RC, CD, and CT. It was also high in the DCT but mild in the PCT. From four weeks until 12 weeks, the localization of the two lectins was the same, and the immunoreactivity increased with age. Conlusions: Our research elucidates the neonatal and postnatal intra-renal localization of ghrelin and jacalin. [ABSTRACT FROM AUTHOR]

The tumor microenvironment that refers to the tumor's surroundings is a key modulator of tumor growth and invasion. The tumor-derived signals are known to downregulate the anti-tumor effects of the effector cells present in the TME. Thus, the cross-talk between the tumor cells with the surrounding immune cells helps in evading the tumor surveillance as well as aiding in tumor growth and proliferation. Hence, knowledge regarding the effects of drugs/compound on the tumor-stromal interactions is gaining importance. In the present study, the effects of jacalin, a dietary lectin on the proliferation and cytokine production of peripheral blood mononuclear cells (PBMCs), are investigated. Jacalin was shown to act as a mitogen of PBMCs, the key cytokine secreting immune cells. Also, jacalin initially induced increased mRNA expression of pro-inflammatory cytokine IFN-γ; however, prolonged stimulation of PBMCs resulted in increased expression of anti-inflammatory cytokine, mainly TGF-β. Furthermore, 6 h jacalin prestimulated PBMCs (Jac-PBMCs) were shown to inhibit HeLa cell proliferation while 24 h Jac-PBMCs were found to favor tumor growth. Thus, it may be postulated that while jacalin initially polarizes the PBMCs to hinder the tumor growth, after a stipulated time point, interaction of jacalin with PBMCs can lead to an immunosuppressive TME that may probably assist in tumor growth and progression. [ABSTRACT FROM AUTHOR]

This study demonstrates the extraction of jacalin from crude extract of jackfruit seeds using anionic surfactant based reverse micellar system, sodium bis(2-ethylhexyl) sulfosuccinate (AOT) in isooctane. Effects of various parameters, such as, NaCl concentration (0.05 – 1 M), pH of aqueous phase (pH 4 – 10), AOT concentration (5 – 150 mM), contact time (5 – 30 min) and phase volume ratio (0.5-5) on the transfer efficiency of jacalin was evaluated by changing one factor at a time (OFAT) while keeping the other parameters constant. A maximum of 83% of protein transfer was achieved after equal volume of organic and aqueous phase was stirred for 20 min using 20 mM AOT at pH 5 aqueous phase containing 0.1 M NaCl. The findings demonstrated AOT reverse micellar system as a promising and effective method to extract and purify jacalin from crude protein mixture.

The genomes of higher plants encode a variety of proteins with lectin domains that are able to specifically recognize certain carbohydrates. Plants are enriched in a variety of potentially complementary glycans, many of which are located in the cell wall. We performed a genome-wide search for flax proteins with lectin domains and compared the expression of the encoding genes in different stem tissues that have distinct cell wall types with different sets of major polysaccharides. Over 400 genes encoding proteins with lectin domains that belong to different families were revealed in the flax genome; three quarters of these genes were expressed in stem tissues. Hierarchical clustering of the data for all expressed lectins grouped the analyzed samples according to their characteristic cell wall type. Most lectins differentially expressed in tissues with primary, secondary, and tertiary cell walls were predicted to localize at the plasma membrane or cell wall. These lectins were from different families and had various architectural types. Three out of four flax genes for proteins with jacalin-like domains were highly upregulated in bast fibers at the stage of tertiary cell wall deposition. The dynamic changes in transcript level of many genes for lectins from various families were detected in stem tissue over the course of gravitropic response induced by plant gravistimulation. The data obtained in this study indicate a large number of lectin-mediated events in plants and provide insight into the proteins that take part in tissue specialization and reaction to abiotic stress.

The potential antitumor effects of jacalin, the plant lectin that specifically recognizes the tumor-associated Thomsen-Friedenreich antigen has been extensively studied. We had earlier reported jacalin to be mitogenic to K562, the Bcr-Abl expressing erythroleukemia cell line. The dearth of studies highlighting the proliferative effects of jacalin and other lectins motivated us to unveil the mechanism underlying the mitogenic effects of jacalin. Caveolin-1 (cav-1) is an integral membrane protein, known to play a crucial role in cell signaling, lipid transport, and membrane trafficking. The role of cav-1 in tumorigenesis is considered to be controversial as it can suppress as well as promote tumor growth, depending on the cellular context. In the present study, we propose that cav-1 plays the central role in the mitogenic effects of jacalin on the K562 cells. In accordance, the mRNA, as well as protein expression of cav-1 was found to be upregulated in the jacalin-treated K562 cells as compared to the untreated control. Further, jacalin stimulation also increased the phosphorylation of ERK and Akt. The rationale that leads to the initial conjecture about cav-1 was that the sequence of jacalin possesses a cav-1-binding site. [ABSTRACT FROM AUTHOR]

The genomes of higher plants encode a variety of proteins with lectin domains that are able to specifically recognize certain carbohydrates. Plants are enriched in a variety of potentially complementary glycans, many of which are located in the cell wall. We performed a genome-wide search for flax proteins with lectin domains and compared the expression of the encoding genes in different stem tissues that have distinct cell wall types with different sets of major polysaccharides. Over 400 genes encoding proteins with lectin domains that belong to different families were revealed in the flax genome; three quarters of these genes were expressed in stem tissues. Hierarchical clustering of the data for all expressed lectins grouped the analyzed samples according to their characteristic cell wall type. Most lectins differentially expressed in tissues with primary, secondary, and tertiary cell walls were predicted to localize at the plasma membrane or cell wall. These lectins were from different families and had various architectural types. Three out of four flax genes for proteins with jacalin-like domains were highly upregulated in bast fibers at the stage of tertiary cell wall deposition. The dynamic changes in transcript level of many genes for lectins from various families were detected in stem tissue over the course of gravitropic response induced by plant gravistimulation. The data obtained in this study indicate a large number of lectin-mediated events in plants and provide insight into the proteins that take part in tissue specialization and reaction to abiotic stress. [ABSTRACT FROM AUTHOR]

In silico analysis revealed that a lectin, jacalin from jackfruit seeds, recognizes a glycosylated region of the receptor-binding domain (RBD) of SARS-CoV2. Jacalin binding induces conformational changes in RBD and significantly affects its interaction with human angiotensin-converting enzyme 2. The result may open up exploration of lectin-based strategies against COVID-19.

ABSTRACT: Advances in the fields of glycobiology and immunology have provided many insights into the role of carbohydrate-protein interactions in the immune system. Jacalin of Artocarpus integrifolia (JCA) and structural mannoprotein of Saccharomyces uvarum (MPS) are molecules with immunomodulatory properties. JCA is an IgA human lectin binding molecule that causes the mitogenic stimulation of immune cells, production of cytokines, chemotaxis, and activation of leukocytes. Studies on the immunomodulatory properties of JCA and MPS in mammals and fish suggest that they have an action on antibody production. The aim of this study was to investigate the possible action of JCA and MPS on the production of specific antibodies in laying hens. For this, laying hens were inoculated with an intra abdominal injection of sheep red blood cells (SRBC) with either JCA (0.075 µg, 0.75 µg, and 7.5 µg) or MPS (20 µg and 100 µg). Levels of anti-SRBC antibodies of the IgY, IgM, and IgA classes were evaluated by ELISA. Results showed that JCA and MPS have immunomodulatory effects on levels of anti-SRBC IgM, IgA, and IgY. An immunostimulatory effect of JCA was observed in primary immune response on anti-SRBC IgY, while an inhibitory effect of JCA and MPS was observed in secondary immune response on the production of IgM and IgA anti-SRBC. These results suggested that MPS and JCA have immunomodulatory effects on antibody production and could be used in future studies on humoral immune response in poultry.

Advances in the fields of glycobiology and immunology have provided many insights into the role of carbohydrate--protein interactions in the immune system. Jacalin of Artocarpus integrifolia (JCA) and structural mannoprotein of Saccharomyces uvarum (MPS) are molecules with immunomodulatory properties. JCA is an IgA human lectin binding molecule that causes the mitogenic stimulation of immune cells, production of cytokines, chemotaxis, and activation of leukocytes. Studies on the immunomodulatory properties of JCA and MPS in mammals and fish suggest that they have an action on antibody production. The aim of this study was to investigate the possible action of JCA and MPS on the production of specific antibodies in laying hens. For this, laying hens were inoculated with an intra abdominal injection of sheep red blood cells (SRBC) with either JCA (0.075 µg, 0.75 µg, and 7.5 µg) or MPS (20 µg and 100 µg). Levels of anti-SRBC antibodies of the IgY, IgM, and IgA classes were evaluated by ELISA. Results showed that JCA and MPS have immunomodulatory effects on levels of anti-SRBC IgM, IgA, and IgY. An immunostimulatory effect of JCA was observed in primary immune response on anti-SRBC IgY, while an inhibitory effect of JCA and MPS was observed in secondary immune response on the production of IgM and IgA anti-SRBC. These results suggested that MPS and JCA have immunomodulatory effects on antibody production and could be used in future studies on humoral immune response in poultry [ABSTRACT FROM AUTHOR]

Jacalin is a major lectin present in jackfruit seeds, obtained by crude protein recovery. Its lectin symbol is AIL and it belongs to the galactose family of N-acetylglucosamine binding lectins, which are recognized to be cancer cell inhibitor. There have been many pharmacological research studies focusing intensively on jacalin, however their scope was restricted to the application of jacalin in pharmacology. Jacalin or lectin extract from jackfruit seed were induced cell cycle arrest in human breast cancer cell, MCF7, in comparison of crude extract, purified jacalin and jacalin standard. IC50 for MCF7 was achieved at concentration 125 µg/mL, comparable to jacalin standard with only 1.60 % contradictions

The cells in the foliose thallus of trebouxiophycean alga Prasiola japonica apparently develop into 2 × 2 cell groups composed of two two-celled groups, each of which is a pair of derivative cells of the latest cell division. In the present study, the structural features of cell walls of the alga P. japonica concerning the formation of the cell groups were investigated using histochemical methods. Thin cell layers stained by Calcofluor White appeared to envelope the two-celled and four-celled groups separately and, hence, separated them from neighboring cell groups, and the Calcofluor White-negative gaps between neighboring four-celled groups were specifically stained by lectins, such as soybean agglutinin, jacalin, and Vicia villosa lectin conjugated with fluorescein. These results indicated that the Calcofluor White-positive cell wall layer of parent cell that existed during two successive cell divisions structurally distinguished two-celled and four-celled groups from others in this alga. Moreover, the results suggested that the cell wall components of the Calcofluor White-negative gaps would possibly contribute to the formation of the planar thallus through lateral union of the cell groups. [ABSTRACT FROM AUTHOR]

Abstract: Ultrathin films of CdSe nannocrystals (CdSe‐NCs) modified with glycosilated molecules have shown interesting electrochemical properties which lead to the detection of lectin Jacalin. [ABSTRACT FROM AUTHOR]

In any therapeutic modality the usage of drug in high doses often leads to serious side-effects. Herein, we demonstrated a method to enhance the antibacterial efficacy of CuS NPs at lower concentration through interacting with jackfruit seed lectin, jacalin. Fluorescence quenching studies revealed that jacalin form complex with CuS NPs and the association constant was 1.91 × 10 4  M −1 . Upon complex with jacalin, the bacterial minimum inhibitory concentration (MIC) of CuS NPs drastically decreases from 12.5 μM to 0.78 μM. The addition of jacalin specific sugar, galactose to jacalin-CuS NPs complex (JCuS NPs) reverses the MIC from 0.78 μM to 25 μM. Mechanistic study suggests that JCuS NPs kills bacteria in part by reactive oxygen species and membrane damage, but galactose prevents the action of JCuS NPs at 0.78 μM. JCuS NPs successfully reduce (14 fold) A. hydrophila colonization in an infected zerbra fish and rescue them completely from the infection, but galJCuS NPs and CuS NPs were ineffective at 0.78 μM. Collectively, our studies demonstrates that the enhance antibacterial activity of JCuS NPs is likely due to the interaction between the galactose binding site of jacalin and the bacterial strains, as a result NPs are targeted and delivered sufficiently. [ABSTRACT FROM AUTHOR]

The irreversible thermal unfolding of jacalin, the lectin purified from jackfruit seeds was accompanied by aggregation, where intermolecular interactions among the subunits are favoured over intramolecular interactions. The extent of aggregation increased as a function of temperature, time and protein concentration. The anionic surfactant, sodium dodecyl sulphate (SDS) significantly suppressed the formation of aggregates as observed by turbidity measurements and Rayleigh scattering assay. Moreover, far UV-CD spectra indicate that the protein β sheet transforms into α helical structure, when denatured in the presence of 3 mM SDS. Further, jacalin when heated in the presence of SDS partially retained the hemagglutination activity when jacalin-SDS mixture was diluted to 1:8 factor since 3 mM SDS was found to lyse the red blood cells. Thus, SDS only altered the aggregation behaviour of jacalin by preventing intermolecular hydrogen bonding among the exposed residues but did not completely stabilize the native conformation. [ABSTRACT FROM AUTHOR]

Surface functionalizations of quantum dots (QDs) are expected to improve their optoelectronic properties and advance their targeting nature, which is highly warranted for application in molecular imaging and biomedical diagnostics. Therefore, an understanding the interaction between surface functionalization agents and QDs is a prerequisite for their application. Herein, we investigated the interaction between cadmium sulfide (CdS) QDs and dietary T-antigen binding lectin (jacalin) isolated from Indian jack fruit seeds. Fluorescence spectroscopy study showed that CdS QDs effectively quenched the intrinsic fluorescence of jacalin. Analysis of fluorescence quenching at a different temperature indicated that the mechanism of interaction is static and a non-radiation energy transfer occurred within the molecules. The obtained binding constant, K a value in the order of 10 4 M −1 at the tested temperature range suggested that the binding affinity between jacalin and CdS QDs is in the same range as those obtained for the interaction of lectin with carbohydrate. Thermodynamic analysis of the binding data, Δ H 0 =−44.24±1.21 kJ mol −1 , Δ S 0 =−60.92±4.10 J mol − 1 K −1 and Δ G 0 =−26.21±0.1 kJ mol −1 suggested that the binding reaction is enthalpy-driven spontaneous process. Hemagglutination activity of jacalin is well preserved even after binding to CdS QDs, indicating that the jacalin-CdS QDs complex can recognize T-antigens of the malignant tissues. To support the claim, we demonstrated selective fluorescence labeling of chronic myeloid leukemia cells, K562 with jacalin-CdS QDs complex. [ABSTRACT FROM AUTHOR]

According to their sugar recognition specificity, plant lectins are proposed as bioactive proteins with potential in cancer treatment and diagnosis. Helja is a mannose-specific jacalin-like lectin from sunflower which was shown to inhibit the growth of certain fungi. Here, we report its recombinant expression in a prokaryotic system and its activity in neurobalstoma cells. Helja coding sequence was fused to the pET-32 EK/LIC, the enterokinase/Ligation-independent cloning vector and a 35 kDa protein was obtained in Escherichia coli representing Helja coupled to thioredoxin (Trx). The identity of this protein was verified using anti-Helja antibodies. This chimera, named Trx-rHelja, was enriched in the soluble bacterial extracts and was purified using Ni+2-Sepharose and D-mannose-agarose chromatography. Trx-rHelja and the enterokinase-released recombinant Helja (rHelja) both displayed toxicity on human SH-SY5Y neuroblastomas. rHelja decreased the viability of these tumor cells by 75% according to the tetrazolium reduction assay, and microscopic analyses revealed that the cell morphology was disturbed. Thus, the stellate cells of the monolayer became spheroids and were isolated. Our results indicate that rHelja is a promising tool for the development of diagnostic or therapeutic methods for neuroblastoma cells, the most common solid tumors in childhood. [ABSTRACT FROM AUTHOR]

The aim of the present study was to analyse Taenia solium metacestode antigens that were derived from the unbound fraction of jacalin affinity chromatography and subsequent tert-octylphenoxy poly (oxyethylene) ethanol Triton X-114 (TX-114) partitioning in the diagnosis of human neurocysticercosis (NCC). Immunoassays were designed to detect T. solium-specific IgG antibodies by ELISA and immunoblot. Serum samples were collected from 132 individuals who were categorised as follows: 40 had NCC, 62 presented Taenia spp or other parasitic diseases and 30 were healthy individuals. The jacalin-unbound (J unbound ) fraction presented higher sensitivity and specificity rates than the jacalin-bound fraction and only this fraction was subjected to subsequent TX-114 partitioning, resulting in detergent (DJ unbound ) and aqueous (AJ unbound ) fractions. The ELISA sensitivity and specificity were 85% and 84.8% for J unbound , 92.5% and 93.5% for DJ unbound and 82.5% and 82.6% for AJ unbound . By immunoblot, the DJ unbound fraction showed 100% sensitivity and specificity and only serum samples from patients with NCC recognised the 50-70 kDa T. solium-specific components. We conclude that the DJ unbound fraction can serve as a useful tool for the differential immunodiagnosis of NCC by immunoblot.

Total antigen from Leishmania (Leishmania) amazonensis and isolates from the Leishmania braziliensis complex, along with their respective antigenic fractions obtained by affinity chromatography on concanavalin-A-Sepharose and jacalin-agarose columns evaluated using immunoenzymatic ELISA assay. For this, serum samples from 229 patients were used, grouped as American tegmental leishmaniasis (nº=58), visceral leishmaniasis (nº=28), Chagas disease (nº=49), malaria (nº=32), tuberculosis (nº=13) and healthy volunteers (nº=49). Samples from American tegmentary leishmaniasis showed higher reactivity with antigens isolated from the Leishmania braziliensis complex than with antigens from Leishmania amazonensis (pAntígeno total de Leishmania (Leishmania) amazonensis e isolado do complexo Leishmania brazilienis, assim como suas respectivas frações antigênicas obtidas por cromatografia de afinidade em coluna de concanavalina-A ligada a sepharose e Jacalina ligada a agarose foram avaliadas por ensaio imunoenzimático ELISA. Para tanto, foram utilizadas amostras de soros de 229 pacientes agrupadas em leishmaniose tegumentar americana (nº=58), leishmaniose visceral (nº=28), doença de Chagas (nº=49), malaria (nº=32), tuberculose (nº=13) e voluntários saudáveis (nº=49). Houve maior reatividade das amostras de leishmaniose tegumentar americana com a utilização dos antígenos obtidos do isolado do complexo Leishmania braziliensis quando comparado com antígenos de Leishmania amazonensis (p

Dietary lectins have been shown to affect the proliferation of human cancer cell lines. The anti-proliferative effects of lectins from varied sources have been extensively studied and in some cases, the underlying mechanism has been explored. Except for peanut agglutinin (PNA), the mitogenic effects of no other lectins have been studied in detail. In the present study, we have shown that jacalin, lectin purified from jackfruit ( Artocarpus integrifolia) seeds act as a mitogen for K562, the Bcr-Abl expressing erythroleukemia cell line (K562) and the effect was found to be dose dependent. K562 cells remained in the proliferative state for a longer period even after the withdrawal of jacalin stimulation, thus jacalin was found to induce sustained mitogenic effect on K562 cells. Further, conditioned media from K562 cells treated with jacalin were observed to have the similar mitogenic effect even in the presence of galactose. Importantly, galactose which is a known ligand for jacalin will interact with functionally active jacalin present in the conditioned media and neutralise its effect. In addition, jacalin treatment also resulted in increased mRNA expression levels of pro-inflammatory cytokines including IL-1β, IL-6 and IFN-γ. Our results indicate that jacalin induces secretion of soluble molecules, which maybe responsible for this observed increased proliferation of K562 cells. [ABSTRACT FROM AUTHOR]

The possibility of early cancer diagnosis has inspired the development of electrochemical systems that could improve both cell detection and activity monitoring, especially for circulating tumor cells. In this study, we propose an impedimetric leukemia sensor based on the use of immobilized jacalin, which is a lectin overexpressed in most types of human cancers. The biosensor is prepared through the chemical adsorption of (3-aminopropil)trimetoxisilano on indium tin oxide electrodes, followed by immersion in a jacalin solution. The performance of the jacalin-modified electrodes toward monocytic leukemic cells, THP-1, and myeloblastic leukemic cells, OCI-AMI3, was investigated through electrochemical impedance spectroscopy. Our results revealed that, upon using the jacalin biorecognition layer, the sensors were able to differentiate leukemic cells from healthy monocyte cells, even at low percentage ranges, with limits of detection of 4±1 cells mL−1 for OCI-AMI3 and 3±1 cells mL−1 for THP-1. [ABSTRACT FROM AUTHOR]

The advances in nanomedicine demonstrate the anticancer properties of silver nanoparticles (AgNPs) and considered as an alternative to the available chemotherapeutic agents. Owing to the preferential interaction of Artocarpus integrifolia lectin (jacalin) with Gal β1-3 GalNAc α (a chemically well-defined tumor associated antigen), a study was undertaken to understand the interaction mechanism of AgNPs with jacalin in presence of specific sugar, galactose. Fluorescence spectroscopic analysis revealed that the AgNPs binding significantly quenched the intrinsic fluorescence of jacalin through a static quenching mechanism, and a non-radiative energy transfer occurred within the molecules. Association constants obtained from the interaction of different sugar-stabilized AgNPs with jacalin are in the order of 10 4 M −1 , this is in the same range as those obtained for the interaction of lectin with carbohydrate and hydrophobic ligand. Each subunit of the tetrameric jacalin binds one AgNPs, and the stoichiometry was unaffected by the presence of the specific sugar, galactose. Hemagglutination assay shows that sugar stabilized AgNPs interacts to jacalin at a site that is different from the saccharide-binding site. Analysis of the FTIR spectra of jacalin indicates that the binding of AgNPs does not alter the secondary structure of jacalin. More importantly, AgNPs exists in nano form even after interacting with the lectin. These results suggest that the development of lectin–AgNPs conjugate would be possible for diagnosis and treatment of cancer. [ABSTRACT FROM AUTHOR]

O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. In proteomic studies, O-glycosylation-site analysis is even more difficult because of the complexity of the sample. In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. Jacalin was used to selectively isolate O-GalNAc glycopeptides before LC-MS-MS analysis, which is optional for individual proteins and necessary for complex human-plasma proteins. Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. The workflow indicated superior sensitivity by not only covering most previously known O-glycosylation sites but also discovering several novel sites. Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. The approach described herein is simple, sensitive, and global for site analysis of core 1 through core 4 O-glycosylated proteins. [ABSTRACT FROM AUTHOR]

Several ruthenium complexes are regarded as anticancer agents and considered as an alternative to the widely used platinum complexes. Owing to the preferential interaction of jacalin with tumor-associated T-antigen, we report the interaction of jacalin with four ruthenium complex namely, tris(1,10-phenanthroline)ruthenium(II)chloride, bis(1,10-phenanthroline)(N-[1,10]phenanthrolin-5-yl-pyrenylmethanimine)ruthenium(II)chloride, bis(1,10-phenanthroline)(dipyrido[3,2- a :2′,3′- c ]-phenazine)ruthenium(II)chloride, bis(1,10-phenanthroline)(11-(9-acridinyl)dipyrido[3,2-a:2′,3′-c]phenazine)ruthenium(II) chloride. Fluorescence spectroscopic analysis revealed that the ruthenium complexes strongly quenched the intrinsic fluorescence of jacalin through a static quenching procedure, and a non-radiative energy transfer occurred within the molecules. Association constants obtained for the interaction of different ruthenium complexes with jacalin are in the order of 10 5 M −1 , which is in the same range as those obtained for the interaction of lectin with carbohydrate and hydrophobic ligand. Each subunit of the tetrameric jacalin binds one ruthenium complex, and the stoichiometry is found to be unaffected by the presence of the specific sugar, galactose. In addition, agglutination activity of jacalin is largely unaffected by the presence of the ruthenium complexes, indicating that the binding sites for the carbohydrate and the ruthenium complexes are different. These results suggest that the development of lectin–ruthenium complex conjugate would be feasible to target malignant cells in chemo-therapeutics. [ABSTRACT FROM AUTHOR]