Differential binding of ppGpp and pppGpp to E-coli RNA polymerase: photo-labeling and mass spectral studies

(p) ppGpp, a secondary messenger, is induced under stress and shows pleiotropic response. It binds to RNA polymerase and regulates transcription in Escherichia coli. More than 25 years have passed since the first discovery was made on the direct interaction of ppGpp with E. coli RNA polymerase. Several lines of evidence suggest different modes of ppGpp binding to the enzyme. Earlier cross-linking experiments suggested that the beta-subunit of RNA polymerase is the preferred site for ppGpp, whereas recent crystallographic studies pinpoint the interface of beta'/omega-subunits as the site of action. With an aim to validate the binding domain and to follow whether tetra-and pentaphosphate guanosines have different location on RNA polymerase, this work was initiated. RNA polymerase was photo-labeled with 8-azido-ppGpp/8-azido-pppGpp, and the product was digested with trypsin and subjected to mass spectrometry analysis. We observed three new peptides in the trypsin digest of the RNA polymerase labeled with 8-azido-ppGpp, of which two peptides correspond to the same pocket on beta'-subunit as predicted by X-ray structural analysis, whereas the third peptide was mapped on the beta-subunit. In the case of 8-azido-pppGpp-labeled RNA polymerase, we have found only one cross-linked peptide from the beta'-subunit. However, we were unable to identify any binding site of pppGpp on the beta-subunit. Interestingly, we observed that pppGpp at high concentration competes out ppGpp bound to RNA polymerase more efficiently, whereas ppGpp cannot titrate out pppGpp. The competition between tetraphosphate guanosine and pentaphosphate guanosine for E. coli RNA polymerase was followed by gel-based assay as well as by a new method known as DRaCALA assay.

Remarkable photo-induced anticancer activity of vitamin-S6 Schiff base copper(II) complexes of pyridyl bases

Vitamin-B6 (VB6) Schiff base (H2L) copper(II) complexes of pyridyl bases, viz. Cu(bpy)(L)] (1), Cu(phen)(L)] (2) and Cu(dppz)(L)] (3), where bpy is 2,2'-bipyridine, phen is 1,10-phenanthroline and dppz is dipyrido3,2-a:2',3'c]phenazine are synthesized, characterized and their phto-induced anticancer activity studied. The non-electrolytic one electron paramagnetic complexes exhibit a d-d band near 700 nm in DMF. The dppz complex intercalatively binds to calf-thymus DNA with binding constant (K-b) values of similar to 10(6) M-1. This complex exhibits low chemical nuclease activity but excellent DNA photocleavage activity when irradiated with red light of 705 nm forming (OH)-O-center dot radical. It displays remarkable photocytotoxicity in human cervical cancer cells (HeLa) giving IC50 value of 0.9 mu M in visible light (400-700 nm) while being less toxic in darkness (IC50 : 23 mu M). The cellular uptake of the complexes seems to be via VB6 transporting membrane carrier mediated diffusion pathway. Photo-induced cell death follows apoptotic pathway involving photo-generated intracellular reactive oxygen species.

Long-period modulated structure and electric-field-induced structural transformation in Na0.5Bi0.5TiO3-based lead-free piezoelectrics

Na0.5Bi0.5TiO3- based lead-free piezoelectrics exhibiting giant piezostrain are technologically interesting materials for actuator applications. The lack of clarity with regard to the structure of the nonpolar phase of this system has hindered the understanding of the structural mechanism associated with the giant piezostrain and other related phenomena. In this paper, we have investigated the structure and field-induced phase transformation behavior of a model system (0.94 - x) Na0.5Bi0.5TiO3-0.06BaTiO(3)-xK(0.5)Na(0.5)NbO(3) (0.0 <= x <= 0.025). A detailed structural analysis using neutron powder diffraction revealed that the nonpolar phase is neither cubic nor a mixture of rhombohedral (R3c) and tetragonal (P4bm) phases as commonly reported in literature but exhibits a long-period modulated structure, which is most probably of the type root 2 x root 2 x n with n = 16. Our results suggest that the giant piezoelectric strain is associated with a field-induced phase transformation of the long-period modulated structure to rhombohedral R3c structure above a critical field. We also demonstrate that the giant piezostrain is lost if the system retains a fraction of the field-induced R3c phase. A possible correlation among depolarization temperature, giant piezostrain, and its electrical fatigue behavior has also been indicated.

Experimental Study of the Earthquake Recurrence Period and the Trend of Post-Seismic Development

In order to study the earthquake recurrence and the characteristics of earthquake series, rupture tests of rock samples and plexiglass samples were made. On rock samples, a number of acoustic emission (AE) and strain measuring points were deployed; the load was one side direct shear. The variation characteristics of AE and strain at different detecting points around the extra large fracture were observed and studied. On plexiglass samples, a series of inclined cracks were prefabricated by a small-scale compressive testing machine. The samples were then loaded on a shockproof platen, when the samples were loaded, the stress intensity factor (SIF) was determined by the laser interferometric technique and shadow optical method of caustics. The fracture conditions such as material toughness around the extra large fracture were also studied. From those experimental results and the theory of fracture mechanics, the earthquake recurrence period and the trend of post-seismic development were studied.

Optical and micro-structural properties of high photoluminescence efficiency InGaN/AlInGaN quantum well structures

We have studied the optical properties of a series of InGaN/AlInGaN 10-period multiple quantum wells (MQW) with differing well thickness grown by metal-organic vapor-phase epitaxy that emit at around 380 nm. The aim of this investigation was to optimise the room temperature internal quantum efficiency, thus the quantum well (QW) thicknesses were accordingly chosen so that the overlap of the electron/hole wave function was maximised. At low temperature, we observed a reduction of the photo luminescence decay time with decreasing well width in line with the theoretical predictions. For a structure with well thicknesses of 1.5 nm, we measured a photoluminescence internal quantum efficiency of 67% at room temperature with a peak emission wavelength of 382 nm. (c) 2006 Elsevier B.V. All rights reserved.

The MBA-LBA I period in the Kourion region : new evidences from Erimi- Laonin tou Porakou (Lemesos, Cyprus)

The evidence from Erimi-Laonin tou Porakou within the Kouris valley settlement system can offer further new data to outline a picture of the development trends from MBA to LBA I periods in the Kourion region. As to this area, such a “transitional” phase could be particularly relevant to better understand the development process of the mature LBA II–III urban centers (in Alassa as well as the most recent discovered in Erimi-Pitharka). In 2008–2009 a focus investigation (an intensive survey, geoperspections and excavations) within the Kouris Valley Project has been addressed to the site of Erimi-Laonin tou Porakou, which lies on one of the highest hilltops facing southward the Kouris dam. The excavations evidenced a double circuit wall which surrounds the top mound and the lower terrace. The top mound (Area A) has been widely occupied by a complex workshop, directly linked with a storage area. The natural limestone bedrock has been worked out to arrange a system of carved deep basins connected each other by a series of flow channels. Southward from the workshop, a large storage area has been cleared, with a relevant assemblage of big pithoi and storage devices. The first lower terrace area (Area B) has been possibly occupied by a domestic quarter; a cemetery area (Area E) has been also evidenced outside the external circuit wall, where a series of five rock-cut tombs (Tombs 228– 232) with single chamber and small incoming dromoi has been excavated.

A possible alchemist apparatus from the early islamic period excavated at Ramla, Israel

Abstract: During extensive salvage excavations carried out during the years 2004-2008 in a large Early Islamic industrial area at the vicinities of Ramla, in Central Israel, an unparalleled industrial device was unearthed. The star-shaped, soil embedded installation, whose lower part was preserved, consisted of a central pottery jar surrounded by five minor jars, linked by ceramic pipes. Evidence of heat was observed mainly around the central vessel, and metal hollow cones perforated in the tip were found inside the surrounding jars. Although the manufacturing procedures and operation techniques of the installation are not completely clear, it is proposed that the installation is part of an industrial workshop or an alchemy laboratory. Both industry and alchemy were well-developed during the Early Islamic period and very often closely related, to the point that sometimes it is difficult to distinguish between them. The identification proposed is based on comparisons with tools described in literary sources, and somewhat later drawings and etchings. Circumstantial ceramic evidence was found, as well as the proximity of a bathhouse whose guests could have been the consumers of perfumes and unguents seem to reinforce this possibility. Due to the poor state of preservation of the device and the lack of available comparisons, the identification proposed here is tentative, and future research coupled with eventual new discoveries is needed in order to clarify this matter.

The Comovement between Monetary and Fiscal Policy Instruments during the Post-War Period in the U.S.

This paper empirically studies the dynamic relationship between monetary and fiscal policies by analyzing the comovements between the Fed funds rate and the primary deficit/output ratio. Simple economic thinking establishes that a negative correlation between Fed rate and deficit arises whenever the two policy authorities share a common stabilization objective. However, when budget balancing concerns lead to a drastic deficit reduction the Fed may reduce the Fed rate in order to smooth the impact of fiscal policy, which results in a positive correlation between these two policy instruments. The empirical results show (i) a significant negative comovement between Fed rate and deficit and (ii) that deficit and output gap Granger-cause the Fed funds rate during the post-Volcker era, but the opposite is not true.

Influence of the northeastern Atlantic oceano-meteorological variability on the northern hake ("merluccius merluccius"). Analysis of the 1978-2006 period

316 p. : il., graf., map.

Influence of Foliar Exposure, Adjuvants, and Rain-free Period on the Efficacy of Glyphosate for Torpedograss Control

The proportion of torpedograss tissue exposed to glyphosate at application rates of 0.28, 0.56, 1.12, 2.24, and 4.48 kg/ha affected control as measured by regrowth. The effect of tissue exposure was more pronounced as application rate decreased. This study suggests that higher rates of glyphosate need to be used during higher water levels, when less torpedograss tissue is exposed to herbicide spray and lower rates may be used during periods of low water levels. Addition of the water conditioning agent Quest (R) (0.25% v/v) to glyphosate spray mixtures diminished the influence of simulated rain events following glyphosate application. Twelve other adjuvants did not influence the effect of simulated rain events.

Northern range extension, abundance and distribution of Pacific coastal Bottlenose doplhins (Tursiops truncatus gilli) in Monterey Bay, California

Pacific coastal bottlenose dolphins (Tursiops truncatus gilli) have apparently moved to Monterey Bay as a result of a shift north of their known range. Between 1983 and 1993, 417 sightings were reported off central California. Eighty-four boat-based surveys, between October 1990 and November 1993, resulted in the photo-identification of 68 uniquely marked individuals. School size ranged between 2 and 35 animals (mean = 16.60, S.D. = 7.72). Forty-three (63%) of the dolphins identified were previously photographed in the Southern California Bight before 1989. Jolly-Seber population estimates indicated an increase in the Monterey Bay population from 1990 to 1993. At least 13 of the photo-identified dolphins were present in Monterey Bay throughout the study period. All but two of the calculated coefficients of association were 0.35, indicating a strong bond among resident animals. The occurrence of an El Niño from January 1992 to the end of 1993 may have affected the number of animals present in the bay: mean school size was significantly greater during El Niño.

Social ecology of bottlenose dophins in Monterey Bay, California

Between 1990 and 1995, Pacific coastal bottlenose dolphins (Tursiops truncatus gillii) were studied using photo-identification during 228 boat-based surveys of the coastal strip (<1 km offshore) between Marina and New Brighton Beach in Monterey Bay (18 km of coastline). The study period encompassed 3 regular (1990, 1991 and 1995) and 3 El Niño years (1992, 1993, 1994). Based on dorsal fin markings, 97 unique individuals were identified. Eighteen animals (19%) showed a high level of site fidelity (defined as presence in at least 5 of the 6 years), although their overall range was larger than the study area. Thirty-eight animals (39%) were transient, leaving for periods of time, and 41 (42%) were occasional encounters. The rate of discovery indicated a pulsed recruitment of new individuals into the study area, with periods of stable school composition, especially during non-El Nino years, and periods of high school fluidity. Encounter rate was significantly higher in El Niño (81%) than non-El Niño years (61%). School size averaged 16 individuals (C.I.3, =0.05) and was significantly larger in El Niño years. Schools where calves were present were twice as large (mean=15; S.D.=8) than schools without calves (mean=8; S.D.=6). Newborns represented 12% of the sightings and were seen year round with a peak in summer and fall. Crude birth rate ranged between 0.09 and 0.17 (mean=0.13; S.D.=0.03). Five females calved in consecutive years and a resident female calved once a year for the duration of the study, possibly indicating a high rate of mortality for calves in this area. Individuals often traveled as subgroups of more consistent composition than the school itself, possibly indicating that a stronger social bond exists within these units which may function as “bands” (sensu Wells 1991) of same sex individuals traveling within a larger school of mixed composition. (ppt file of poster)