On the rotational brownian motion of a bacterial idle motor. II. Theory of fluorescence correlation spectroscopy

The photon flux autocorrelation function of a fluorescent label attached to a bacterial motor shaft is calculated for the case in which the bacterial motor is considered to be actively but idly rotating. It is shown that even when the fluorescent label has a very short lifetime, fluorescence correlation spectroscopy should provide a useful tool for determining the rate of revolution of the bacterial motor under various solution conditions.     

No More Foreigners? The Remaking of German Naturalization and Citizenship Law, 1990–2000

Since the beginning of the 1990s, Germany has been taking controversial policy measures that had been quite unimaginable, even in the 1980s. On the one hand, regulations for asylum seekers and ethnic Germans have gradually become restrictive in nature. On the other hand, changes to the Citizenship and Nationality Law of 1913 (Reichs und Staatsangehörigkeitsgesetz) and the Alien Act of 1965 (Ausländergesetz) have dramatically altered the naturalization process. Before the passage of the new German citizenship law in 1999, the acquisition of citizenship in Germany was solely based on the principle of descent (jus sanguinis). This paper attempts to explain why German government altered its citizenship policy and introduced birthright citizenship. It reviews the post-war contextual factors that have shaped the debate on citizenship and asks whether it is external or internal factors, that is international-level norms and institutions or domestic politics, that led to the change. If domestic politics can fully explain the adoption of the new citizenship law, what domestic forces played an important role? Was state action taken in the name of protecting “the national interest” or was it party politics? What role did societal forces play in the making of new citizenship policy? Did the act seek to mitigate anti-foreigner sentiment growing in Germany since the 1990s? Using archival and documentary research and secondary data on socio-economic and demographic trends in Germany, this paper concludes that domestic politics explain the changes in the citizenship law. I argue that granting and withholding national citizenship, including birthright citizenship, is exclusively in the hands of a nation state. Considering a purely ethnocultural understanding of nation-hood now includes legal and territorial components, the new German citizenship law is a major reform at the both theoretical and conceptual level. However, the practical consequences of the new naturalization process remain to be seen. The main question for the future research agenda is whether the new naturalization and citizenship laws will bring full integration of foreigners living in German society.     

Correlation between the parameters of contingent negative variation and characteristics of variational pulsometry in Parkinsonian patients

In patients suffering from Parkinson's disease (PD), we analyzed correlations between the parameters of contingent negative variation (CNV) and data of variational pulsometry (according to the measurements of R-R ECG intervals). Studies were carried out on 35 patients (group PD), 49 to 74 years old, with the stage of disease of 1.5 to 3.0 according to the Hoehn-Yahr international classification. In the course of CNV recording (i.e., in the state of a certain functional loading), we observed significant negative correlations between the integral magnitude (area) of this potential and indices of variational pulsometry (RMSSD, SDNN, C. var, and HF) that characterize the intensity of parasympathetic (respiratory) influences on the cardiovascular system. In the control group, such correlations were absent. We found significant correlations between the autonomic balance, CNV magnitude, and stage of PD reflecting the level of generalization of the pathological process. In the subgroup of patients with the PD stage 1.5 to 2.0, significant changes in the mean values of indices of parasympathetic influences during recording of the CNV were not observed, while in another subgroup (the PD stage 2.5 to 3.0), these values increased significantly (P < 0.05 and P < 0.01). If the estimates of the PD stage were low, the CNV area demonstrated greater values (P < 0.01). The disturbance of coordination of muscle-to-muscle interactions in the PD group is, probably, an important factor responsible for parasympathetic dysregulation and suppression of the CNV generation. We found positive correlation between the intensity of parasympathetic influences in the course of CNV recording and the level of postural disorders (r = 0.37, P < 0.05). On the contrary, the CNV magnitude demonstrated a negative correlation with the intensity of these disorders (r = −0.36, P < 0.05), as well as with the level of postural instability (r = −0.55, P < 0.001). We hypothesize that alterations of the autonomic balance and the activity of those cerebral structures, which are responsible for the motor readiness, result, to a significant extent, from weakening of the activity of the noradrenergic system due to degenerative processes developing in cells of the locus coeruleus. The impairment of the latter structure, together with degeneration of neurons of the substantia nigra and a decrease in the level of nigro-striatal dopamine, underlies the pathomorphological pattern of PD. Neirofiziologiya/Neurophysiology, Vol. 40, No. 3, pp. 242–253, May–June, 2008.     

Bayesian Nonparametric Modeling for Comparison of Single-Neuron Firing Intensities

Summary .  We propose a fully inferential model-based approach to the problem of comparing the firing patterns of a neuron recorded under two distinct experimental conditions. The methodology is based on nonhomogeneous Poisson process models for the firing times of each condition with flexible nonparametric mixture prior models for the corresponding intensity functions. We demonstrate posterior inferences from a global analysis, which may be used to compare the two conditions over the entire experimental time window, as well as from a pointwise analysis at selected time points to detect local deviations of firing patterns from one condition to another. We apply our method on two neurons recorded from the primary motor cortex area of a monkey's brain while performing a sequence of reaching tasks.     

Module-based construction of plasmids for chromosomal integration of the fission yeast Schizosaccharomyces pombe

Integration of an external gene into a fission yeast chromosome is useful to investigate the effect of the gene product. An easy way to knock-in a gene construct is use of an integration plasmid, which can be targeted and inserted to a chromosome through homologous recombination. Despite the advantage of integration, construction of integration plasmids is energy- and time-consuming, because there is no systematic library of integration plasmids with various promoters, fluorescent protein tags, terminators and selection markers; therefore, researchers are often forced to make appropriate ones through multiple rounds of cloning procedures. Here, we establish materials and methods to easily construct integration plasmids. We introduce a convenient cloning system based on Golden Gate DNA shuffling, which enables the connection of multiple DNA fragments at once: any kind of promoters and terminators, the gene of interest, in combination with any fluorescent protein tag genes and any selection markers. Each of those DNA fragments, called a ‘module’, can be tandemly ligated in the order we desire in a single reaction, which yields a circular plasmid in a one-step manner. The resulting plasmids can be integrated through standard methods for transformation. Thus, these materials and methods help easy construction of knock-in strains, and this will further increase the value of fission yeast as a model organism.     

The forensic use of luminol chemiluminescence to detect traces of blood inside motor vehicles.

The luminol test for blood was carried out on a set of interior fittings and surfaces inside three different makes of modern motor car. The surfaces and fittings provided little interference with the test for blood, although there was some detectable chemiluminescence when the test was applied to blood-free material from a seatbelt, a boot-lining and a gear-knob. The case with which haemoglobin samples could be washed off interior car surfaces was also examined for seat fabrics, carpets, roof-linings and various other plastic interior surfaces. A standard wash with water alone was not very effective and removed only ca. 50% of the haemoglobin. A standard wash with soapy water or with a proprietary multipurpose car cleaner removed ca. 90% of the haemoglobin from the tested surface. The effect of high car interior temperatures on haemoglobin samples that were subsequently used in the luminol test was also examined. It was shown that the sensitivity of the luminol test was not decreased but was increased by the prior heating of a haemoglobin sample. This effect was attributed to the thermal conversion of haemoglobin to the more brighter catalyst for chemiluminescence, methaemoglobin. The enthalpy of this conversion in the solid state was found to be 14.1 kJ/mol.     

Mutations in the E1a gene of type 5 adenovirus result in oncogenic transformation of Fischer rat embryo cells

Transformation of a specific clone of Fischer rat embryo (CREF) cells with wild-type 5 adenovirus (Ad5) or the E1a plus E1b transforming gene regions of Ad5 results in epithelioid transformants that grow efficiently in agar but that do not induce tumors when inoculated into nude mice or syngeneic Fischer rats. In contrast, CREF cells transformed by a host-range Ad5 mutant, H5hrl, which contains a single base-pair deletion of nucleotide 1055 in E1a resulting in a 28-kd protein (calculated) in place of the wild-type 51-kd acidic protein, display a cold-sensitive transformation phenotype and an incomplete fibroblastic morphology but surprisingly do induce tumors in nude mice and syngeneic rats. Tumors develop in both types of animals following injection of CREF cells transformed by other cold-sensitive Ad5 E1a mutants (H5dl101 and H5in106), which contain alterations in their 13S mRNA and consequently truncated 289AA proteins. CREF cells transformed with only the E1a gene (0-4.5 m.u.) from H5hrl or H5dl101 also produce tumors in these animals. To directly determine the role of the 13S E1a encoded 289AA protein and the 12S E1a encoded 243AA protein in initiating an oncogenic phenotype in adenovirus-transformed CREF cells, we generated transformed cell lines following infection with the Ad2 mutant pm975, which synthesizes the 289AA E1a protein but not the 243AA protein, and the Ad5 mutant H5dl520 and the Ad2 mutant H2dl1500, which do not produce the 289AA E1a protein but synthesize the normal 243AA E1a protein. All three types of mutant adenovirus-transformed CREF cells induced tumors in nude mice and syngeneic rats. Tumor formation by these mutant adenovirus-transformed CREF cells was not associated with changes in the arrangement of integrated adenovirus DNA or in the expression of adenovirus early genes. These results indicate, therefore, that oncogenic transformation of CREF cells can occur in the presence of a wild-type 13S E1a protein or a wild-type 12S E1a protein when either protein is present alone, but does not occur when both wild-type E1a proteins are present.     

Physiological integration in Cassia fasciculata Michx.: inflorescence removal and defoliation experiments

Summary In the annual herb Cassia fasciculata virtually every leaf subtends an axillary inflorescence. We examined the degree to which these leaf-inflorescence units (reproductive nodes) were physiologically independent of each other in the production of flowers, fruits, and seeds. Removal of up to 4 of every 5 inflorescences resulted in substantial increases in fruit and seed production by remaining, intact reproductive nodes. These increases nearly compensated for and manipulated reproductive nodes were associated with different vascular strands. When 2 of every 3 leaves were removed, fruit and seed production were reduced at both intact and defoliated reproductive nodes. Taken together, these results suggest that neighboring reproductive nodes in C. fasciculata are not physiologically independent of one another, and that competition among fruits and seeds for parental resources occurs over several reproductive nodes.Scientific contribution no. 1595 from the New Hampshire Agricultural Experiment Station     

Fast axon activity and the motor pattern in cockroach legs during swimming

Abstract Electromyographic recordings were made from muscles that extend the trochanter/femur of each of the six legs of American cockroaches, Periplaneta americana (L.), while the insects swam in water. The recordings showed two novel features. (1) During swimming, muscle activity in different legs was coordinated in the alternating tripod pattern commonly seen during free walking on land, not in the pattern of synchronous leg pairs common to other large terrestrial insects in water. (2) Fast axons were usually recruited along with slow axons, even when the insect swam at a moderate pace. Fast axon activity always started after the middle of the slow axon burst in intact insects, but vanished from most bursts in the stump of the leg after amputation of the femur. The alternating tripod pattern was maintained even after amputation. Possible causes of fast axon recruitment are discussed.     

Efficient integrative transformation of the phytopathogenic fungus Alternaria alternata mediated by the repetitive rDNA sequences

An attempt was made to transform Alternaria alternata protoplasts using a plasmid vector, pDH25, bearing the Escherichia coli hygromycin B (Hy) phosphotransferase gene (hph) under the control of the Aspergillus nidulans trpC promoter. Transformants arose on a selective medium containing 100 μg Hy/ml. There were two types of transformants, forming large and small colonies on the selective medium. Transformation with one μg of the vector produced an average of 4.5 large colonies and 600 small ones. In large-colony transformants, the vector often integrated into the recipient chromosome in the form of highly rearranged tandem arrays. To increase transformation efficiency, fragments of the highly repetitive ribosomal RNA gene cluster (rDNA) of A. alternata were used to construct four new vectors for homologous recombination system. Use of these vectors gave higher transformation efficiency than the original plasmid. The best vector, pDH25r1a, gave rise to large-colony transformants at a frequency 20 times higher than pDH25. Transformation events in A. alternata with pDH25r1a occured by homologous recombination as a single crossover between the plasmid-borne rDNA segment and its homologue in the chromosome, often giving rise to tandemly repeated vector DNA.