Inhibition of protein synthesis leading to unfolded protein response is the major event in abrin-mediated apoptosis

Abrin obtained from the plant Abrus precatorius inhibits protein synthesis and also triggers apoptosis in cells. Previous studies from our laboratory suggested a link between these two events. Using an active site mutant of abrin A-chain which exhibits 225-fold lower protein synthesis inhibitory activity than the wild-type abrin A-chain, we demonstrate in this study that inhibition of protein synthesis induced by abrin is the major factor triggering unfolded protein response leading to apoptosis. Since abrin A-chain requires the B-chain for internalization into cells, the wild-type and mutant recombinant abrin A-chains were conjugated to native ricin B-chain to generate hybrid toxins, and the toxic effects of the two conjugates were compared. The rate of inhibition of protein synthesis mediated by the mutant ricin B-rABRA (R167L) conjugate was slower than that of the wild-type ricin B-rABRA conjugate as expected. The mutant conjugate activated p38MAPK and caspase-3 similar to its wild-type counterpart although at later time points. Overall, these results confirm that inhibition of protein synthesis is the major event contributing to abrin-mediated apoptosis.

Evidence of episodic brittle faulting in the cratonic part of the Peninsular India and its implications for seismic hazard in slow deforming regions

The region around Waclakkancheri, in the province of Kerala, India, which lies in the vicinity of Palghat-Cauvery ;hear zone (within the Precambrian crystalline terrain), has been a site of microseismic activity since 1989. Earlier studies had identified a prominent WNW-ESE structure overprinting on the E-W trending lineaments associated with Palghat-Cauvery shear zone. We have mapped this structure, located in a chamockite quarry near Desamangalam, Waclakkancheri, which we identify as a ca. 30 km-long south dipping reverse fault. This article presents the characteristics of this fault zone exposed on the exhumed crystalline basement and discusses its significance in understanding the earthquake potential of the region. This brittle deformation zone consists of fracture sets with small-scale displacement and slip planes with embedded fault gouges. The macroscopic as well as the microscopic studies of this fault zone indicate that it evolved through different episodes of faulting in the presence of fluids. The distinct zones within consolidated gouge and the cross cutting relationship of fractures indicate episodic fault activity. At least four faulting episodes can be recognized based on the sequential development of different structural elements in the fault rocks. The repeated ruptures are evident along this shear zone and the cyclic behavior of this fault consists of co-seismic ruptures alternating with inter-seismic periods, which is characterized by the sealed fractures and consolidated gouge. The fault zone shows a minimum accumulated dip/oblique slip of 2.1 m in the reverse direction with a possible characteristic slip of 52 cm (for each event). The ESR dating of fault gouge indicates that the deformation zone records a major event in the Middle Quaternary. The empirical relationships between fault length and slip show that this fault may generate events M >= 6. The above factors suggest that this fault may be characterized as potentially active. Our study offers some new pointers that can be used in other slow deforming cratonic hinterlands in exploring the discrete active faults.

The 26 July 2005 heavy rainfall event over Mumbai: numerical modeling aspects

The performance of the Advanced Regional Prediction System (ARPS) in simulating an extreme rainfall event is evaluated, and subsequently the physical mechanisms leading to its initiation and sustenance are explored. As a case study, the heavy precipitation event that led to 65 cm of rainfall accumulation in a span of around 6 h (1430 LT-2030 LT) over Santacruz (Mumbai, India), on 26 July, 2005, is selected. Three sets of numerical experiments have been conducted. The first set of experiments (EXP1) consisted of a four-member ensemble, and was carried out in an idealized mode with a model grid spacing of 1 km. In spite of the idealized framework, signatures of heavy rainfall were seen in two of the ensemble members. The second set (EXP2) consisted of a five-member ensemble, with a four-level one-way nested integration and grid spacing of 54, 18, 6 and 1 km. The model was able to simulate a realistic spatial structure with the 54, 18, and 6 km grids; however, with the 1 km grid, the simulations were dominated by the prescribed boundary conditions. The third and final set of experiments (EXP3) consisted of a five-member ensemble, with a four-level one-way nesting and grid spacing of 54, 18, 6, and 2 km. The Scaled Lagged Average Forecasting (SLAF) methodology was employed to construct the ensemble members. The model simulations in this case were closer to observations, as compared to EXP2. Specifically, among all experiments, the timing of maximum rainfall, the abrupt increase in rainfall intensities, which was a major feature of this event, and the rainfall intensities simulated in EXP3 (at 6 km resolution) were closest to observations. Analysis of the physical mechanisms causing the initiation and sustenance of the event reveals some interesting aspects. Deep convection was found to be initiated by mid-tropospheric convergence that extended to lower levels during the later stage. In addition, there was a high negative vertical gradient of equivalent potential temperature suggesting strong atmospheric instability prior to and during the occurrence of the event. Finally, the presence of a conducive vertical wind shear in the lower and mid-troposphere is thought to be one of the major factors influencing the longevity of the event.

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.

Coronopilin-another major sesquiterpene lactone in Parthenium hysterophorus

No abstract is available.

Microprocessor-based event sequence recorder

An event sequence recorder is a specialized piece of equipment that accepts inputs from switches and contactors, and prints the sequence in which they operate. This paper describes an event sequence recorder based on an Intel 8085 microprocessor. It scans the inputs every millisecond and prints in a compact form the channel number, type of event (normal or abnormal) and time of occurrence. It also communicates these events over an RS232C link to a remote computer. A realtime calendar/clock is included. The system described has been designed for continuous operation in process plants, power stations etc. The system has been tested and found to be working satisfactorily.

K-Ar ages of biotite and muscovite from Pangong metamorphic complex, Shyok suture zone, India: Implications for the youngest post-collision metamorphic event in Ladakh Himalaya

The garnet-kyanite-staurolite and garnet-biotite-staurolite gneisses were collected from a locality within Lukung area that belongs to the Pangong metamorphic complex in Shyok valley, Ladakh Himalaya. The kyanite-free samples have garnet and staurolite in equilibrium, where garnets show euhedral texture and have flat compositional profile. On the other hand, the kyanite-bearing sample shows equilibrium assemblage of garnet-kyanite-staurolite along with muscovite and biotite. In this case, garnet has an inclusion rich core with a distinct grain boundary, which was later overgrown by inclusion free euhedral garnet. Garnet cores are rich in Mn and Ca, while the rims are poor in Mn and rich in Fe and Mg, suggesting two distinct generations of growth. However, the compositional profiles and textural signature of garnets suggests the same stage of P -T evolution for the formation of the inclusion free euhedral garnets in the kyanite-free gneisses and the inclusion free euhedral garnet rims in the kyanite-bearing gneiss. Muscovites from the four samples have consistent K-Ar ages, suggesting the cooling age (∼ 10 Ma) of the gneisses. These ages make a constraint on the timing of the youngest post-collision metamorphic event that may be closely related to an activation of the Karakoram fault in Pangong metamorphic complex.

Design and Performance Evaluation of EXMAN: An EXtended MANchester Data Flow Computer

Data flow computers are high-speed machines in which an instruction is executed as soon as all its operands are available. This paper describes the EXtended MANchester (EXMAN) data flow computer which incorporates three major extensions to the basic Manchester machine. As extensions we provide a multiple matching units scheme, an efficient, implementation of array data structure, and a facility to concurrently execute reentrant routines. A simulator for the EXMAN computer has been coded in the discrete event simulation language, SIMULA 67, on the DEC 1090 system. Performance analysis studies have been conducted on the simulated EXMAN computer to study the effectiveness of the proposed extensions. The performance experiments have been carried out using three sample problems: matrix multiplication, Bresenham's line drawing algorithm, and the polygon scan-conversion algorithm.

Triassic eclogite from northern Vietnam: inferences and geological significance

The first finding of low-temperature eclogites from the Indochina region is reported. The eclogites occur along the Song Ma Suture zone in northern Vietnam, which is widely regarded as the boundary between the South China and Indochina cratons. The major lithology of the area is pelitic schist that contains garnet and phengite with or without biotite, chloritoid, staurolite and kyanite, and which encloses blocks and lenses of eclogite and amphibolite. The eclogites commonly consist of garnet, omphacite, phengite, rutile, quartz and/or epidote with secondary barroisite. Omphacite is commonly surrounded by a symplectite of Na-poor omphacite and Na-rich plagioclase. In highly retrograded domains, diopside + tremolite + plagioclase symplectites replace the primary phases. Estimated peak-pressure metamorphic conditions based on isochemical phase diagrams for the eclogites are 2.1-2.2 GPa and 600-620 degrees C, even though thermobarometric results yield higher pressure and temperature conditions (2.6-2.8 GPa and 620-680 degrees C). The eclogites underwent a clockwise P-T trajectory with a post-peak-pressure increase of temperature to a maximum of > 750 degrees C at 1.7 GPa and a subsequent cooling during decompression to 650 degrees C and 1.3 GPa, which was followed by additional cooling before close-to-isothermal decompression to similar to 530 degrees C at 0.5 GPa. The surrounding pelitic schist (garnet-chloritoid-phengite) records similar metamorphic conditions (580-600 degrees C at 1.9-2.3 GPa) and a monazite chemical age of 243 +/- 4 Ma. A few monazite inclusions within garnet and the cores of some zoned monazite in garnet-phengite schist record an older thermal event (424 +/- 15 Ma). The present results indicate that the Indochina craton was deeply (> 70 km) subducted beneath the South China craton in the Triassic. The Silurian cores of monazite grains may relate to an older non-collisional event in the Indochina craton.

Seismic damage estimation through measurable dynamic characteristics

Ductility based design of reinforced concrete structures implicitly assumes certain damage under the action of a design basis earthquake. The damage undergone by a structure needs to be quantified, so as to assess the post-seismic reparability and functionality of the structure. The paper presents an analytical method of quantification and location of seismic damage, through system identification methods. It may be noted that soft ground storied buildings are the major casualties in any earthquake and hence the example structure is a soft or weak first storied one, whose seismic response and temporal variation of damage are computed using a non-linear dynamic analysis program (IDARC) and compared with a normal structure. Time period based damage identification model is used and suitably calibrated with classic damage models. Regenerated stiffness of the three degrees of freedom model (for the three storied frame) is used to locate the damage, both on-line as well as after the seismic event. Multi resolution analysis using wavelets is also used for localized damage identification for soft storey columns.

Japanese Encephalitis Virus Utilizes the Canonical Pathway To Activate NF-kappa B but It Utilizes the Type I Interferon Pathway To Induce Major Histocompatibility Complex Class I Expression in Mouse Embryonic Fibroblasts

Flaviviruses have been shown to induce cell surface expression of major histocompatibility complex class I (MHC-I) through the activation of NF-kappa B. Using IKK1(-/-), IKK2(-/-), NEMO-/-, and IKK1-/- IKK2-/- double mutant as well as p50(-/-) RelA(-/-) cRel(-/-) triple mutant mouse embryonic fibroblasts infected with Japanese encephalitis virus (JEV), we show that this flavivirus utilizes the canonical pathway to activate NF-kappa B in an IKK2- and NEMO-, but not IKK1-, dependent manner. NF-kappa B DNA binding activity induced upon virus infection was shown to be composed of RelA: p50 dimers in these fibroblasts. Type I interferon (IFN) production was significantly decreased but not completely abolished upon virus infection in cells defective in NF-kappa B activation. In contrast, induction of classical MHC-I (class 1a) genes and their cell surface expression remained unaffected in these NF-kappa B-defective cells. However, MHC-I induction was impaired in IFNAR(-/-) cells that lack the alpha/beta IFN receptor, indicating a dominant role of type I IFNs but not NF-kappa B for the induction of MHC-I molecules by Japanese encephalitis virus. Our further analysis revealed that the residual type I IFN signaling in NF-kappa B-deficient cells is sufficient to drive MHC-I gene expression upon virus infection in mouse embryonic fibroblasts. However, NF-kappa B could indirectly regulate MHC-I expression, since JEV-induced type I IFN expression was found to be critically dependent on it.

Primary Structural Documentation of the Major Urinary Protein of the Indian Commensal Rat (Rattus rattus) Using a Proteomics Platform

Purpose: A number of proteome studies have been performed recently to identify pheromone-related protein expression and their molecular function using genetically modified rodents' urine. However, no such studies have used Indian commensal rodents; interestingly, in a previous investigation, we confirmed the presence of volatile molecules in commensal rodents urine and these molecules seem to be actively involved in pheromonal communication. Therefore, the present study aims to identify the major urinary protein [MUP] present in commensal rat urine, which will help us to understand the protein's expression pattern and intrinsic properties among the rodents globally. Experimental Design: Initially, the total urinary proteins were separated by 1-D and 2-D electrophoresis and then subsequently analyzed by Matrix Assisted Laser Desorption Ionization-Time of Flight and Mass Spectrometer (MALDI-TOF/MS). Furthermore, they were then fragmented with the aid of a Tandem Mass Spectrometer (TOF/TOF) and the identified sequences aligned and confirmed using similarity with the deduced primary structures of members of the lipocalin superfamily.Results: The SDS-PAGE protein profiles showed distinct proteins with molecular masses of 15, 22.4, 25, 28, 42, 50, 55, 68, and 91 kDa. Of these proteins, the 22.4 kDa protein was considered to be target candidate. When 2D electrophoresis was carried out, about similar to 50 spots were detected with different masses and various pI ranges. The 22.4 kDa protein was found to have a pI of about 4.9. This 22.4 kDa protein spot was digested and subjected to mass spectrometry; it was identified as rat MUP. The fragmented peptides from the rat MUP at 935, 1026, 1192, and 1303 m/z were further fragmented with the aid of MS/MS and generated de novo sequence and this confirmed this protein to be the MUP present in the urine of commensal rats.Conclusion: The present investigation confirms the presence of MUP with a molecular mass of 22.4 kDa in the urine of commensal rats. This protein may be involved in the binding of volatile molecules and opens up a discussion about how volatile and non-volatile molecules in the commensal rats' urine may contribute chemo-communication.

Two-dimensional fuzzy fault tree analysis for chlorine release from a chlor-alkali industry using expert elicitation

The hazards associated with major accident hazard (MAN) industries are fire, explosion and toxic gas releases. Of these, toxic gas release is the worst as it has the potential to cause extensive fatalities. Qualitative and quantitative hazard analyses are essential for the identification and quantification of these hazards related to chemical industries. Fault tree analysis (FTA) is an established technique in hazard identification. This technique has the advantage of being both qualitative and quantitative, if the probabilities and frequencies of the basic events are known. This paper outlines the estimation of the probability of release of chlorine from storage and filling facility of chlor-alkali industry using FTA. An attempt has also been made to arrive at the probability of chlorine release using expert elicitation and proven fuzzy logic technique for Indian conditions. Sensitivity analysis has been done to evaluate the percentage contribution of each basic event that could lead to chlorine release. Two-dimensional fuzzy fault tree analysis (TDFFTA) has been proposed for balancing the hesitation factor involved in expert elicitation. (C) 2010 Elsevier B.V. All rights reserved.

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.

Formation of a G-quadruplex at the BCL2 major breakpoint region of the t(14;18) translocation in follicular lymphoma

The t(14;18) translocation in follicular lymphoma is one of the most common chromosomal translocations. Most breaks on chromosome 18 are located at the 3'-UTR of the BCL2 gene and are mainly clustered in the major breakpoint region (MBR). Recently, we found that the BCL2 MBR has a non-B DNA character in genomic DNA. Here, we show that single-stranded DNA modeled from the template strand of the BCL2 MBR, forms secondary structures that migrate faster on native PAGE in the presence of potassium, due to the formation of intramolecular G-quadruplexes. Circular dichroism shows evidence for a parallel orientation for G-quadruplex structures in the template strand of the BCL2 MBR. Mutagenesis and the DMS modification assay confirm the presence of three guanine tetrads in the structure. 1H nuclear magnetic resonance studies further confirm the formation of an intramolecular G-quadruplex and a representative model has been built based on all of the experimental evidence. We also provide data consistent with the possible formation of a G-quadruplex structure at the BCL2 MBR within mammalian cells. In summary, these important features could contribute to the single-stranded character at the BCL2 MBR, thereby contributing to chromosomal fragility.