B- and T-cell epitopes of tropomyosin, the major shrimp allergen

Seafood allergy is often encountered on ingestion of crustaceans such as shrimp, lobster, crab, and crayfish (1). On eating cooked shrimp, sensitive individuals experience a wide spectrum of reactions ranging from abdominal discomfort to anaphylaxis. The presence of cross-reacting heat-stable allergens in crustacean food was first recognized by Hoffman et al. (2) and Lehrer et al. (3). Subsequently, the major allergen was isolated and characterized from the shrimp species Paneaus indicus (Pen i 1) (4) and I? aztecm (Pen a 1) (5). Pen i 1 (originally designated Sa-TI) and Pen a 1, with mol. mass of 34 and 36 kDa, respectively, contain 301 and 312 amino-acid residues with a predominance of gluta- mate/glutamine and asparatate/asparagine.

Triploid plants from endosperm cultures of sandalwood by experimental embryogenesis

Embryogenesis has been induced from endosperm callus cultures of sandalwood (Santalum album L.). Viable plantlets developed from the embryoids on subculture to White's basal medium supplemented with 0.5 mg/l of indole acetic acid. Chromosomal analysis of the root tips showed the triploid number 3n = 30.

Isolation and characterization of heat-stable allergens from shrimp (Penaeus indicus)

Shrimp are among the more common causes of immediate hypersensitivity reactions to food. To characterize better the allergenic substances within shrimp, extracts from heated shrimp were systematically examined with solid-phase radioimmunoassay and sera from patients clinically sensitive to shrimp. Two heat-stable protein allergens, designated as Sa-I and Sa-II, were identified from boiled shrimp (Penaeus indicus) extracts. Sa-I was isolated by ultrafiltration, Sephadex G-25, and diethylaminoethyl-Sephacel chromatography, whereas Sa-II, the major allergen, was purified by successive chromatography on diethylaminoethyl-Sephacel, Bio-Gel P-200, and Sepharose 4B columns. Sa-I, which was homogeneous by polyacrylamide gel electrophoresis (PAGE), elicited a single band on sodium dodecyl sulfate-PAGE corresponding to a molecular weight of 8.2 kd. Sa-II was also found to be homogeneous by PAGE, crossed immunoelectrophoresis, and immunoblotting. On sodium dodecyl sulfate-PAGE, it elicited a single band with a molecular weight of 34 kd. Sa-II was found to contain 301 amino acid residues and was particularly rich in glutamate/glutamine and aspartate/asparagine. Solid-phase radioimmunoassay-inhibition studies revealed that Sa-I and Sa-II share 54% of the allergenic epitopes, suggesting that Sa-I may be a fragment of Sa-II.SDS-PAGE, Sodium dodecyl sulfate-polyacrylamide gel electrophoresis; MW, Molecular weight; BSA, Bovine serum albumin; DEAE, Diethylaminoethyl; SPRIA, Solid-phase radioimmunoassay; CIE, Crossed immunoelectrophoresis .

Identification of tropomyosin as the major shrimp allergen and characterization of its IgE-binding epitopes

The major heat-stable shrimp allergen (designated as Sa-II), capable of provoking IgE-mediated immediate type hypersensitivity reactions after the ingestion of cooked shrimp, has been shown to be a 34-kDa heat- stable protein containing 300 amino acid residues. Here, we report that a comparison of amino acid sequences of different peptides generated by proteolysis of Sa-II revealed an 86% homology with tropomyosin from Drosophila melanogaster, suggesting that Sa-II could be the shrimp muscle protein tropomyosin. To establish that Sa-II is indeed tropomyosin, the latter was isolated from uncooked shrimp (Penaeus indicus) and its physicochemical and immunochemical properties were compared with those of Sa-II. Both tropomyosin and Sa-II had the same molecular mass and focused in the isoelectric pH range of 4.8 to 5.4. In the presence of 6 M urea, the mobility of both Sa-II and shrimp tropomyosin shifted to give an apparent molecular mass of 50 kDa, which is a characteristic property of tropomyosins. Shrimp tropomyosin bound to specific IgE antibodies in the sera of shrimp-sensitive patients as assessed by competitive ELISA inhibition and Western blot analysis. Tryptic maps of both Sa-II and tropomyosin as obtained by reverse phase HPLC were superimposable. Dot-blot and competitive ELISA inhibition using sera of shrimp-sensitive patients revealed that antigenic as well as allergenic activities were associated with two peptide fractions. These IgE-binding tryptic peptides were purified and sequenced. Mouse anti-anti-idiotypic antibodies raised against Sa-II specific human idiotypic antibodies recognized not only tropomyosin but also the two allergenic peptides, thus suggesting that these peptides represent the major IgE binding epitopes of tropomyosin. A comparison of the amino acid sequence of shrimp tropomyosin in the region of IgE binding epitopes (residues 50-66 and 153-161) with the corresponding regions of tropomyosins from different vertebrates confirmed lack of allergenic cross-reactivity between tropomyosins from phylogenetically distinct species.

Synthesis, spectral characterization, in-vitro microbiological evaluation and cytotoxic activities of novel macrocyclic bis hydrazone

A macrocyclic hydrazone Schiff base was synthesized by reacting 1,4-dicarbonyl phenyl dihydrazide with 2,6-diformyl-4-methyl phenol and a series of metal complexes with this new Schiff base were synthesized by reaction with Co(II), Ni(II) and Cu(II) metal salts. The Schiff base and its complexes have been characterized by elemental analyses, IR, H-1 NMR, UV-vis, FAB mass, ESR spectra, fluorescence, thermal, magnetic and molar conductance data. The analytical data reveal that the Co(II), Ni(II) and Cu(II) complexes possess 2:1 metal-ligand ratios. All the complexes are non-electrolytes in DMF and DMSO due to their low molar conductance values. Infrared spectral data suggest that the hydrazone Schiff base behaves as a hexadentate ligand with NON NON donor sequence towards the metal ions. The ESR spectral data shows that the metal-ligand bond has considerable covalent character. The electrochemical behavior of the copper(II) complex was investigated by cyclic voltammetry. The Schiff base and its complexes have also been screened for their antibacterial (Escherichia coli, Staphylococcus aureus, Shigella dysentery, Micrococcus, Bacillus subtilis, Bacillus cereus and Pseudomonas aeruginosa) and antifungal activities (Aspergillus niger, Penicillium and Candida albicans) by MIC method. The brine shrimp bioassay was also carried out to study their in-vitro cytotoxic properties. (C) 2009 Elsevier Masson SAS. All rights reserved.

Induction of allergen-specific IgE and IgG responses by anti-idiotypic antibodies

In order to explore idiotypic, anti-idiotypic, and anti-anti-idiotypic responses to allergens, BALB/c mice were immunized with affinity- purified human idiotypic antibodies directed against a highly purified shrimp allergen. This resulted in the production of anti-idiotypic antibodies which were quantitated by using rabbit idiotypic antibodies raised against the same purified allergen. The mouse anti-idiotypic antibodies recognized shrimp-specific human idiotypic antibodies of the IgE isotype from 18 of 20 individuals, and IgG antibodies from 14 of 20 shrimp-sensitive patients. Immunization of BALB/c mice with affinity- purified, allergen-specific anti-idiotypic antibodies induced anti- allergen IgE and IgG responses in the absence of the allergen. This paper thus presents evidence that anti-idiotypic antibodies raised against allergen-specific idiotypic antibodies may substitute for the original allergen in the induction of allergen-specific idiotypic antibodies. The demonstration of shared idiotopes on IgG and IgE antibodies in the sera of shrimp-sensitive patients supports the use of allergen-specific anti-idiotypic antibodies as surrogate allergens.

Demonstration of IgE Antibodies to Nucleic Acid Antigens in Patients with Sle

Using a combination of avidin-biotin microELISA and solid phase radioimmunoassay, we examined sera from 23 patients with systemic lupus erythematosus (SLE), two patients with established sensitivity to ingested shrimp, and 15 healthy normal subjects. In addition to IgG antibodies, varying amounts of IgE antibodies specific for native DNA (nDNA), denatured or single-stranded DNA (dnDNA), RNA, and tRNA were demonstrable in the sera of SLE patients, but not in the sera of normal subjects. A comparison of the specificity of nucleic acid-specific IgE antibodies present in the sera of shrimp-sensitive patients with those present in the sera of seven SLE patients revealed that the IgE antibodies in the sera of shrimp-sensitive patients specifically recognized shrimp tRNA but not yeast tRNA, calf thymus RNA, or calf thymus DNA, while those present in the sera of patients with SLE recognized all these nucleic acid antigens. The IgE antibodies directed against nDNA, dnDNA, RNA, and tRNA may mediate mast cell and basophil degranulation and thus contribute both to immediate-type hypersensitivity phenomena including hives seen in patients with SLE and to the localization of IgE-nucleic acid complexes in target

On the dynamics of activation of mammalian X chromosomes

We have investigated a mathematical model of the process of activation of the X chromosomes in eutherian mammals. The model assumes that the activation is brought about over some definite time interval T by the complete saturation of N receptor sites on an X chromosome by M activating molecules (or multiples of M). The probability λ of a first hit on the receptor site is considered to be very much lower than that of subsequent hits; that is, we assume strong co-operative binding. Assuming further that an incomplete saturation of receptor sites is malfunctional, we can show that for proper activation of X chromosomes in normal diploid males and females, we must have λMT ≥ 3 and 0·96 ≤ N/M ≤ 1. An extension of this analysis for the triploid cases shows that under these conditions, we cannot explain the activation of two X's if the number of activating molecules is fixed at M. This suggests that there must be two classes of triploid embryos differing from each other in a step-wise manner in the number of activating molecules. In other words, triploids with two active X chromosomes would require 2M activating molecules as opposed to M molecules in triploids with a single active X. This interpretation of the two classes of triploids would be consistent with differing imprinting histories of the parental contributions to the triploid zygote.

Chronicle of Marine Diatom Culturing Techniques

Diatoms are regarded as useful neutral lipid sources, as liquid fuel precursors, as foods for marine culture of zooplankters, larval and post-larval shrimp, copepods, juvenile oysters and as micromachines in nanotechnology. Combining microscopic observation with in situ culturing has been useful in areas of taxonomy, ecology, biomonitoring, biotechnology, etc. This communication reviews various culturing techniques of marine diatoms with the relative merits.

Caryophyllene-Rich Essential Oil of Didymocarpus tomentosa: Chemical Composition and Cytotoxic Activity

The essential oil from the leaves of Didymocarpus tomentosa was extracted by hydrodistillation and analyzed by GC/FID and GC/MS. Twenty five constituents amounting to 81.6% of the oil were identified. The leaf oil contained 78.7% sesquiterpenes and 2.9% monoterpenes. The leaf essential oil of D. tomentosa is a unique caryophyllene-rich natural source containing beta-caryophyllene, caryophyllene oxide, alpha-humulene and humulene oxide. The cytotoxic activity of the oil was determined by the BSLT using shrimp larva and the MTT assay using HeLa tumor cell line. The oil showed significant cytotoxic activity with LC50 and IC50 values of 12.26 and 11.4 mu g/mL, respectively. This is the first report on the chemical composition and cytotoxic activity of the essential oil of D. tomentosa.

A Rodinian suture in western India: New insights on India-Madagascar correlations

We report detailed evidence for a new paleo-suture zone (the Kumta suture) on the western margin of southern India. The c. 15-km-wide, westward dipping suture zone contains garnet-biotite, fuchsite-haematite, chlorite-quartz, quartz-phengite schists, biotite augen gneiss, marble and amphibolite. The isochemical phase diagram estimations and the high-Si phengite composition of quartz-phengite schist suggest a near-peak condition of c. 18 kbar at c. 550 degrees C, followed by near-isothermal decompression. The detrital SHRIMP U-Pb zircon ages from quartz-phengite schist give four age populations ranging from 3280 to 2993 Ma. Phengite from quartz-phengite schist and biotite from garnet-biotite schist have K-Ar metamorphic ages of ca. 1326 and ca. 1385 Ma respectively. Electron microprobe-CHIME ages of in situ zircons in quartz-phengite schist (ca. 3750 Ma and ca. 1697 Ma) are consistent with the above results. The Bondla ultramafic-gabbro complex in the west of the Kumta suture compositionally represents an arc with K-Ar biotite ages from gabbro in the range 1644-1536 Ma. On the eastern side of the suture are weakly deformed and unmetamorphosed shallow westward-dipping sedimentary rocks of the Sirsi shelf, which has the following upward stratigraphy: pebbly quartzite/sandstone, turbidite, magnetite iron formation, and limestone; farther east the lower lying quartzite has an unconformable contact with ca. 2571 Ma quartzo-feldspathic gneisses of the Dharwar block with a ca. 1733 Ma biotite cooling age. To the west of the suture is a c. 60-km-wide Karwar block mainly consisting of tonalite-trondhjemite-granodiorite (TTG) and amphibolite. The TTGs have U-Pb zircon magmatic ages of ca. 3200 Ma with a rare inherited core age of ca. 3601 Ma. The K-Ar biotite cooling age from the TTGs (1746 Ma and 1796 Ma) and amphibolite (ca. 1697 Ma) represents late-stage uplift. Integration of geological, structural and geochronological data from western India and eastern Madagascar suggest diachronous ocean closure during the amalgamation of Rodinia; in the north at around ca. 1380 Ma, and a progression toward the south until ca. 750 Ma. Satellite imagery based regional structural lineaments suggests that the Betsimisaraka suture continues into western India as the Kumta suture and possibly farther south toward a suture in the Coorg area, representing in total a c. 1000 km long Rodinian suture. (C) 2013 Elsevier B.V. All rights reserved.

Neoarchean arc magmatism followed by high-temperature, high-pressure metamorphism in the Nilgiri Block, southern India

The Nilgiri Block, southern India is an exhumed lower crust formed through arc magmatic processes in the Neoarchean. The main lithologies in this terrane include charnockites, gneisses, volcanic tuff, metasediments, banded iron formation and mafic-ultramafic bodies. Mafic-ultramafic rocks are present towards the northern and central part of the Nilgiri Block. We examine the evolution of these mafic granulites/metagabbros by phase diagram modeling and U-Pb sensitive high resolution ion microprobe (SHRIMP) dating. They consist of a garnet-clinopyroxene-plagioclase-hornblende-ilmenite +/- orthopyroxene +/- rutile assemblage. Garnet and clinopyroxene form major constituents with labradorite and orthopyroxene as the main mineral inclusions. Labradorite, identified using Raman analysis, shows typical peaks at 508 cm(-1), 479 cm(-1), 287 cm(-1) and 177 cm(-1). It is stable along with orthopyroxene towards the low-pressure high-temperature region of the granulite fades (M1 stage). Subsequently, orthopyroxene reacted with plagioclase to form the peak garnet + clinopyroxene + rutile assemblage (M2 stage). The final stage is represented by amphibolite facies-hornblende and plagioclase-rim around the garnet-clinopyroxene assemblage (M3 stage). Phase diagram modeling shows that these mafic granulites followed an anticlockwise P-T-t path during their evolution. The initial high-temperature metamorphism (M1 stage) was at 850-900 degrees C and similar to 9 kbar followed by high-pressure granulite fades metamorphism (M2 stage) at 850-900 degrees C and 14-15 kbar. U-Pb isotope studies of zircons using SHRIMP revealed late Neoarchean to early paleoproterozoic ages of crystallization and metamorphism respectively. The age data shows that these mafic granulites have undergone arc magmatism at ca. 25392 +/- 3 Ma and high-temperature, high-pressure metamorphism at ca. 2458.9 +/- 8.6 Ma. Thus our results suggests a late Neoarchean arc magmatism followed by early paleoproterozoic high-temperature, high-pressure granulite facies metamorphism due to the crustal thickening and suturing of the Nilgiri Block onto the Dharwar Craton. (C) 2015 Elsevier B.V. All rights reserved.