Circular Dichroism Studies of the Structure of DNA Complex with Berberine

Abstract

The binding of the benzodioxolo-benzoquinolizine alkaloid, berberine chloride to natural and synthetic DNAs has been studied by intrinsic and extrinsic circular dichroic measurements. Binding of berberine causes changes in the circular dichroism spectrum of DNA as shown by the increase of molar ellipticity of the 270nm band, but with very little change of the 240nm band. The molar ellipticity at the saturation depends strongly on the base composition of DNA and also on salt concentration, but always larger for the AT rich DNA than the GC rich DNA The features in the circular dichroic spectral changes of berberine-synthetic DNA complexes were similar to that of native DNA but depends on the sequence of base pairs.

On binding to DNA and polynucleotides, the alkaloid becomes optically active. The extrinsic circular dichroism developed in the visible absorption region (300–500nm) for the berberine-DNA complexes shows two broad spectral bands in the regions 425–440nm and 340–360nm with the maximum varying depending on base composition and sequence of DNA While the 425nm band shows less variation on the binding ratio, the 360nm band is remarkably dependent on the DNA/alkaloid ratio. The generation of the alkaloid associated extrinsic circular dichroic bands is not dependent on the base composition or sequence of base pairs, but the nature and magnitude of the bands are very much dependent on these two factors and also on the salt concentration. The interpretation of the results with respect to the modes of the alkaloid binding to DNA are presented.     

Primary cilia and forebrain development

With a microtubule-based axoneme supporting its plasma membrane-ensheathed projection from the basal body of almost all cell types in the human body, and present in only one copy per cell, the primary cilium can be considered an organelle sui generis. Although it was first observed and recorded in histological studies from the late 19th century, the tiny structure was essentially forgotten for many decades. In the past ten years, however, scientists have turned their eyes once again upon primary cilia and realized that they are very important for the development of almost all organs in the mammalian body, especially those dependent upon the signaling from members Hedgehog family, such as Indian and Sonic hedgehog. In this review, we outline the roles that primary cilia play in forebrain development, not just in the crucial transduction of Sonic hedgehog signaling, but also new results showing that cilia are important for cell cycle progression in proliferating neural precursors. We will focus upon cerebral cortex development but will also discuss the importance of cilia for the embryonic hippocampus, olfactory bulb, and diencephalon.     

Amplitudes and time scales of picosecond-to-microsecond motion in proteins studied by solid-state NMR: a critical evaluation of experimental approaches and application to crystalline ubiquitin

Solid-state NMR provides insight into protein motion over time scales ranging from picoseconds to seconds. While in solution state the methodology to measure protein dynamics is well established, there is currently no such consensus protocol for measuring dynamics in solids. In this article, we perform a detailed investigation of measurement protocols for fast motions, i.e. motions ranging from picoseconds to a few microseconds, which is the range covered by dipolar coupling and relaxation experiments. We perform a detailed theoretical investigation how dipolar couplings and relaxation data can provide information about amplitudes and time scales of local motion. We show that the measurement of dipolar couplings is crucial for obtaining accurate motional parameters, while systematic errors are found when only relaxation data are used. Based on this realization, we investigate how the REDOR experiment can provide such data in a very accurate manner. We identify that with accurate rf calibration, and explicit consideration of rf field inhomogeneities, one can obtain highly accurate absolute order parameters. We then perform joint model-free analyses of 6 relaxation data sets and dipolar couplings, based on previously existing, as well as new data sets on microcrystalline ubiquitin. We show that nanosecond motion can be detected primarily in loop regions, and compare solid-state data to solution-state relaxation and RDC analyses. The protocols investigated here will serve as a useful basis towards the establishment of a routine protocol for the characterization of ps–μs motions in proteins by solid-state NMR.     

Circadian Locomotor Rhythms in Mice with Targeted Disruption of the Gene for the Carbon Monoxide Synthesizing Enzyme,Heme Oxygenase-2

Carbon monoxide (CO), generated in neurons by the enzyme heme oxygenase-2 (HO2), is postulated to be a gaseous signaling molecule in the mammalian brain. Because of the recent evidence suggesting an important role of another endogenously produced gas, nitric oxide (NO), in entrainment of circadian rhythms in mammals, we hypothesized that CO may also be involved in regulating these rhythms. Consistent with this idea, others have found a circadian rhythm of heme turnover and CO synthesis can be induced by bright light. Furthermore, HO2 is co-localized with guanylyl cyclase, the putative target of CO, throughout the brain, with high amounts of staining in the suprachiasmatic nucleus (SCN) of the hypothalamus. The goal of the present study was to evaluate the role of CO in photic entrainment in wild-type and HO2 deficient mice. HO2–/– mice did not display any abnormalities in circadian rhythmicity. Entrainment to a light–dark cycle, the ability to phase delay locomotor activity after a four hour phase shift in photoperiod, and the period of the free running rhythm (t) were similar between HO2–/– and wild-type mice. Taken together, these data suggest that CO does not play a major role in regulating circadian activity rhythms in mice.     

Properties and Nematicide Performance of Avermectins

In 1979, the anthelmintic activity of abamectin, a mixture of avermectins B_1a and B_1b, was first reported. Since then, multiple articles have investigated avermectins' degradation and its efficacy against a wide variety of pests under different conditions and using different modes of application. However, there is a gap in the literature of analysing abamectin properties and its performance as a non‐fumigant nematicide when applied liquid or granular vs. new avenues of application based on seed and seedling treatment. Therefore, this article reviewed literature to discuss the mode of action, environmental aspects, the nematicidal effectiveness of treatment forms and the range of activity to address these topics.     

Seed structure, germination, and reserve mobilization in Butia capitata (Arecaceae)

Butia capitata is a palm tree endemic to the Cerrado biome of Brazil and has significant potential for ornamental and food uses. In this work, we characterized the structures of the seeds and seedlings of this species to identify anatomical aspects related to its pronounced dormancy and determine the processes involved in reserve mobilization. Intact seeds, and seeds from which the operculum had been removed, were allowed to germinate and their morphology, physiology, anatomy, and histochemistry, together with those of the seedlings, were followed for 30 days. The seed coat was found to be rich in phenolic compounds and not lignified. The endosperm contains abundant protein and lipidic reserves, and the embryo has additional starch reserves. Germination occurred only in seeds with their opercula removed and involved the elongation of the cotyledon cells and meristematic activity in the “M zone” located between the embryonic axis and the proximal extremity of the embryo. The mobilization of embryonic reserves initiates during the first phase of imbibition, while the mobilization of endosperm reserves represents a post-germination event associated with the formation of a secretory epidermis and aerenchyma and the vascularization of the haustorium. Seeds with intact opercula did not germinate, but demonstrated embryonic reserve mobilization and cell elongation, indicating that dormancy in B. capitata is related to the incapacity of the embryo to dislocate the operculum.     

Orientation of European corn borer first instar larvae to synthetic green leaf volatiles

European corn borer (ECB) neonate larvae are capable of orienting towards maize odours and of avoiding spinach odours. We previously reported that maize odours’ attraction was dependent on the stimulus regime. This led us to propose that maize odours could have a repellent or attractive effect depending on their concentration. In this work, we tested this hypothesis by evaluating attraction or avoidance of neonate ECB larvae to four concentrations of each of six single green leaf volatiles (GLVs); these are commonly found in maize and other plants. We found a dose‐dependent effect for all of these GLVs with the exception of 1‐hexyl acetate, which did not elicit any orientation behaviour over the range of concentrations tested. These five GLVs were repellent at high concentrations, while two of them were attractive at a lower concentration. These observations indicate for the first time that plant odours induce different behaviours in ECB neonate larvae depending not only on their chemical identity but also their concentration.     

Current Status of Cereal Root Diseases in Western Australia Under Intensive Cereal Production and Their Comparison with the Historical Survey Conducted During 1976–1982

Two separate surveys of root diseases of cereals in the Western Australian (WA) cereal belt were conducted: the first conducted annually for wheat and barley during 1976–1982 and the second for wheat during 2005–2007. For the 1976–1982 survey, the cereal belt was divided into 15 zones based on the location and rainfall. Sampling was representative of the actual cropping area, with both wheat and barley sampling sites selected by zone as a percentage of total sites. Over 31 000 plants were assessed from a total of 996 fields. Average take‐all incidence ranged from 3% in the northern low rainfall zone to 57% in the southern high rainfall zone. Other root diseases assessed included rhizoctonia root rot, fusarium crown rot and subcrown internode discolouration. During the 2005–2007 survey, around 20 000 plants from a total of 210 fields being intensively cropped with cereals were surveyed for take‐all, rhizoctonia root rot, fusarium crown rot, common root rot, root lesion nematode and cereal cyst nematode. The 2005–2007 survey results indicated that root and crown diseases prevailed in paddocks frequently cropped with cereals and occurred at damaging levels across all WA cropping districts surveyed. The more recent root disease survey identified that the fungal diseases rhizoctonia root rot and fusarium crown rot and the root lesion nematode were the most serious impediments to intensive cereal production, particularly in the southern region of WA. Comparing the 2005–2007 results with the previous survey of 1976–1982, the relative importance of take‐all appears to have declined over the past 30 years.     

A Laboratory study on the visual and chemical orientation of the gastropod Nerita Fulgurans Gmelin, 1791

Behavioural responses of the gastropod Nerita fulgurans Gmelin, 1791 to flat black rectangles and intraspecific mucus trails were measured in a circular arena. Snails were tested in water either in the presence or absence of chemicals generated from a predator gastropod, Chicoreus brevifrons (Lamarck, 1822). The test hypothesis was that this snail has different behavioural responses as result of visual and chemical cue integration. Nerita fulgurans has the capacity to orient to solid targets subtending angles larger than 10° and follow its own mucus trails. In water conditioned by the predator C . brevifrons, snails exhibited an avoidance response when 10°, 20° and 45° sectors were presented, demonstrating an integration of chemical and visual information. The simultaneous presentation of two orienting cues (black sectors and mucus trails) was tested to determine the nature of the interaction. When the two cues were oriented in the same direction there was no effect. When the two cues were presented from directions 180° apart a preference for visual cues over mucus trail cues was evident when the visual angle of the visual cue subtended angles greater than 90°. This result demonstrates a hierarchical usage of the orienting references.