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In a subset of papillary renal cell carcinomas a t(X;1)(p11;q21)chromosome translocation has repeatedly been reported.Positional cloning has demonstrated that,as a result of this translocation,the transcription factor TFE3 gene on the X-chromosome becomes fused to a novel gene,PRCC,on chro- mosome 1.Since as yet little is known about the function of PRCC,we sought to identify the mouse counterpart of the PRCC gene.Isolation and sequence analysis of a mouse Prcc cDNA revealed a high level of conservation between man and mouse,both at the nucleotide and protein level.As the human PRCC gene,the mouse Prcc gene is ubiquitously expressed.It shows low expression in all mouse fetal tissues examined.In addition,we identified a genomic cosmid clone containing the complete Prcc gene.The mouse Prcc gene consists of seven exons,all of which contain coding sequences.The small second exon,which was found to be located adjacent to the t(X;1) breakpoint in the human gene on chromosome 1,is also con- served between man and mouse.In mouse,Prcc is located on chromosome 3.These cDNA and genomic clones will be instru- mental in the creation of mouse models for a further elucida- tion of the function of PRCC. [ABSTRACT FROM AUTHOR]

Pond snails ( Lymnaea stagnalis (L.)) were treated with 2,2′-dichlorobiphenyl (DCB) or 4,4′-DCB, to examine the toxicokinetic profile of these compounds. Snails were treated orally with 210 μg 4,4′-DCB (impregnated on food) for 14 hr, or snails were injected with 50 μg of 2,2′-DCB or 4,4′-DCB in the foot. At different times after starting feeding or injection, tissues (albumen gland, digestive gland and digestive tube, central nervous system, remainder parts), hemolymph and faeces were analyzed for unchanged 2,2′- or 4,4′-DCB. The results showed that in case of oral administration of 4,4′-DCB after 144 hr, 97.5% of the dose was excreted unchanged in the faeces. During the first 48 hr 4,4′-DCB was found in all tissues. Thereafter, an exponential elimination was found (the rate constant of elimination (k) varied from 0.010-0.021 per hr, t from 33-60 hr and the apparent clearance from 0.02-0.3 g/hr for the different tissues). After injection, the compounds were found in all the above mentioned tissues, especially in the digestive gland. There was a clear difference between snails injected with 2,2′- and 4,4′-DCB. Firstly, k for 2,2′-DCB was higher (0.028 per hr vs 4,4′-DCB: 0.001 per hr). Secondly, 2,2′-DCB was lethal; 63% of the animals died after 72 hr. [ABSTRACT FROM AUTHOR]

An important factor for the acceptance and thus the spread of automated and connected driving (ACD) is the degree of subjective uncertainty that users experience when interacting with automated vehicles. Subjective uncertainties always occur when people are not able to predict the further course of a situation or future events due to lack of experience or information. If such uncertainties occur during the use of automated vehicles, the development of trust and thus acceptance of this technology is impaired by the negative emotions accompanying subjective uncertainties. Within the AutoAkzept project (which full title translates to: Automation without uncertainty to increase the acceptance of automated and connected driving), solutions for user-focused automation have been developed that put vehicle occupants at the center of system development. User-focused systems take into account two basic human needs in human-machine interaction, the need to understand and the need to be understood. For this purpose, user-focused systems use different sensors to detect subjective uncertainties and their influencing factors in real time, integrate this information with context data and make adjustments that reduce subjective uncertainties. The systemic adaptations of user-focused systems follow a holistic approach that includes the levels of vehicle guidance, interior adaptation and information presentation as well as target guidance are included. By reducing or avoiding subjective uncertainties, the project developments contribute to a positive, comfortable user experience and help to increase the acceptance of ACD. This paper presents research results of AutoAkzept on the topics of user state and activity modelling as well as needs-based adaptation strategies, which represent key components for the implementation of user-focused automation., (© The Author(s) 2021.)

As the desire for deploying automated ("driverless") vehicles increases, there is a need to understand how they might communicate with other road users in a mixed traffic, urban, setting. In the absence of an active and responsible human controller in the driving seat, who might currently communicate with other road users in uncertain/conflicting situations, in the future, understanding a driverless car's behaviour and intentions will need to be relayed via easily comprehensible, intuitive and universally intelligible means, perhaps presented externally via new vehicle interfaces. This paper reports on the results of a questionnaire-based study, delivered to 664 participants, recruited during live demonstrations of an Automated Road Transport Systems (ARTS; SAE Level 4), in three European cities. The questionnaire sought the views of pedestrians and cyclists, focussing on whether respondents felt safe interacting with ARTS in shared space, and also what externally presented travel behaviour information from the ARTS was important to them. Results showed that most pedestrians felt safer when the ARTS were travelling in designated lanes, rather than in shared space, and the majority believed they had priority over the ARTS, in the absence of such infrastructure. Regardless of lane demarcations, all respondents highlighted the importance of receiving some communication information about the behaviour of the ARTS, with acknowledgement of their detection by the vehicle being the most important message. There were no clear patterns across the respondents, regarding preference of modality for these external messages, with cultural and infrastructural differences thought to govern responses. Generally, however, conventional signals (lights and beeps) were preferred to text-based messages and spoken words. The results suggest that until these driverless vehicles are able to provide universally comprehensible externally presented information or messages during interaction with other road users, they are likely to contribute to confusing and conflicting interactions between these actors, especially in a shared space setting, which may, therefore, reduce efficient traffic flow., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Cyclodextrin glycosyltransferase (CGTase) preferably catalyzes transglycosylation reactions, whereas many other alpha-amylase family enzymes are hydrolases. Despite the availability of three-dimensional structures of several transglycosylases and hydrolases of this family, the factors that determine the hydrolysis and transglycosylation specificity are far from understood. To identify the amino acid residues that are critical for the transglycosylation reaction specificity, we carried out error-prone PCR mutagenesis and screened for Bacillus circulans strain 251 CGTase mutants with increased hydrolytic activity. After three rounds of mutagenesis the hydrolytic activity had increased 90-fold, reaching the highest hydrolytic activity ever reported for a CGTase. The single mutation with the largest effect (A230V) occurred in a residue not studied before. The structure of this A230V mutant suggests that the larger valine side chain hinders substrate binding at acceptor subsite +1, although not to the extent that catalysis is impossible. The much higher hydrolytic than transglycosylation activity of this mutant indicates that the use of sugar acceptors is hindered especially. This observation is in favor of a proposed induced-fit mechanism, in which sugar acceptor binding at acceptor subsite +1 activates the enzyme in transglycosylation [Uitdehaag et al. (2000) Biochemistry 39, 7772-7780]. As the A230V mutation introduces steric hindrance at subsite +1, this mutation is expected to negatively affect the use of sugar acceptors. Thus, the characteristics of mutant A230V strongly support the existence of the proposed induced-fit mechanism in which sugar acceptor binding activates CGTase in a transglycosylation reaction.

The (X;1)(p11;q21) translocation is a recurrent chromosomal abnormality in a subset of human papillary renal cell carcinomas, and is sometimes the sole cytogenetic abnormality present. Via positional cloning, we were able to identify the genes involved. The translocation results in a fusion of the transcription factor TFE3 gene on the X chromosome to a novel gene, designated PRCC, on chromosome 1. Through this fusion, reciprocal translocation products are formed, which are both expressed in papillary renal cell carcinomas. PRCC is ubiquitously expressed in normal adult and fetal tissues and encodes a putative protein of 491 aa with a relatively high content of prolines. No relevant homologies with known sequences at either the DNA or the protein level were found.

A combination of Southern blot analysis on a panel of tumor-derived somatic cell hybrids and fluorescence in situ hybridization (FISH) techniques was used to map a series of DNA markers relative to the 1q21 breakpoint of the renal cell carcinoma (RCC)-associated (X;1)-(p11;q21) translocation. This breakpoint maps between several members of the S100 family which are clustered in the 1q21 region and a conserved region between man and mouse containing the markers SPTA1-CRP-APCS-FcER1A-ATP1A2-APOA2. The location of the breakpoint coincides with the transition of a region of synteny of human chromosome 1 with mouse chromosomes 3 and 1.

The muscle-myosin heavy-chain (mMHC) gene of Drosophila hydei has been sequenced completely (size 23.3 kb). The sequence comparison with the D. melanogaster mMHC gene revealed that the exon-intron pattern is identical. The protein coding regions show a high degree of conservation (97%). The alternatively spliced exons (3a-b, 7a-d, 9a-c, 11a-e, and 15a-b) display more variations in the number of nonsynonymous and synonymous substitutions than the common exons (2, 4, 5, 6, 8, 10, 12, 13, 14, 16, 17, and 19). The base composition at synonymous sites of fourfold degenerate codons (third position) is not biased in the alternative exons. In the common exons there exists a bias for C and against A. These findings imply that the alternative exons of the Drosophila mMHC gene evolve at a different, in several cases higher, rate than the common ones. The 5' splice junctions and 5' and 3' untranslated regions show a high level of similarity, indicating a functional constraint on these sequences. The intron regions vary considerably in length within one species, but the corresponding introns are very similar in length between the two species and all contain stretches of sequence similarity. A particular example is the first intron, which contains multiple regions of similarity. In the conserved regions of intron 12 (head-tail border) sequences were found which have the potential to direct another smaller mMHC transcript.

We have used fluorescence in situ hybridization to map the positions of the different repetitive DNA sequences from the region forming the lampbrush loop pair Nooses on the Y chromosome of Drosophila hydei. This region harbours a megabase cluster of tandemly organized repeats of the Y-specific ay1 family and a megabase cluster of tandem repeats of the related Y-specific YsI family. In addition, ay1 repeats also occur in short blocks that are interspersed by other repetitive DNA sequences that we call Y-associated, since they have additional copies on other chromosomes. Using specific probes for ay1, YsI and Y-associated DNA sequences, we show that there is one large proximal cluster of YsI repeats and one, more distally located, large cluster of ay1 repeats. The Y-chromosomal copies of the Y-associated sequences are located in the most distal part of the ay1 cluster. This is consistent with the juxtaposition of ay1 and Y-associated sequences in more than 300 kb of cloned genomic DNA. Since both ay1 and Y-associated sequences have been shown to be transcribed in the Nooses, the lampbrush loop is formed in a distal region of the short arm of the Y chromosome, adjacent to the terminally located nucleolus organizer region. The clusters of homogeneous ay1 and YsI repeats are of no functional significance for the formation of the lampbrush loop.

In their ultradian (2- to 3-hr) feeding rhythm, common voles show intraindividual synchrony from day to day, as well as interindividual synchrony between members of the population, even at remote distances. This study addresses the question of how resetting of the ultradian rhythm, a prerequisite for such synchronization, is achieved. Common voles were subjected to short light-dark cycles (1 hr darkness with light varying between 0.7 and 2.5 hr); to T cycles (long light-dark cycles in the circadian range--16 hr darkness and 3-13 hr light); to light pulses (15 min) during different circadian and ultradian phases; and to addition of D2O to the drinking water (25%). Short light-dark cycles and D2O were also applied to voles without circadian rhythmicity, after lesions of the suprachiasmatic nuclei. In these experiments, four hypotheses on synchronization of ultradian rhythmicity were tested: (I) synchronization by a direct response to light; (II) synchronization via the circadian system with multiple triggers, here called "cogs," each controlling a single ultradian feeding bout; and (III and IV) synchronization via the circadian system with a single "cog," which resets an ultradian oscillator and either (III) originates directly from the circadian pacemaker, or (IV) is mediated via the overt circadian activity rhythm. Short light-dark cycles failed to entrain ultradian rhythms, either in circadian-rhythmic or in non-circadian-rhythmic voles; light pulses did not cause phase shifts; and in extreme T cycles no stable phase relationship with light could be demonstrated. Thus, Hypothesis I was rejected. Changes in the circadian period (tau) were generated as aftereffects of light pulses, by entrainment in various T cycles, and by the addition of D2O to the drinking water. These changes in tau did not lead to parallel, let alone proportional, changes in the ultradian period. This excluded Hypothesis II. Both in T-cycle experiments and in the D2O experiments with circadian-rhythmic voles, the phase of ultradian feeding bouts was locked to the end of circadian activity rather than to the most prominent marker of the pacemaker, the onset of circadian activity. This was not expected under Hypothesis III, but was consistent with entrainment via activity (Hypothesis IV). On the basis of these experiments, we conclude that the most likely mechanism of ultradian entrainment is that of a light-insensitive ultradian oscillator, reset every dawn by the termination of the activity phase controlled by the circadian pacemaker, which is itself entrained by the light-dark cycle. Neither in circadian-rhythmic nor in non-circadian-rhythmic voles was the period of the feeding rhythm lengthened by administration of D2O. This insensitivity to deuterium is exceptional among biological rhythms.

1. The occurrence of an as yet unidentified cytochrome P-450 in the microsomal fraction of the digestive gland of the snail, Lymnaea stagnalis was studied. 2. Studies in vivo and in vitro (digestive gland homogenates or the 170,000g fraction) of the cytochrome P-450-mediated metabolism of substrates such as biphenyl, pentoxy- and ethoxy-resorufin and aminopyrine have been made. 3. Cytochrome P-450 concentration in the digestive gland calculated from CO-difference spectra was 0.30 +/- 0.05 nmol/g tissue. This amount was not increased either by phenobarbital or by 3-methylcholanthrene. 4. Aniline binding spectra resembled normal type II spectra, while type I model substrates such as hexobarbital and 2,2'-dichlorobiphenyl showed type II- or reversed type I-like spectra. 5. O-Deethylation of 7-ethoxyresorufin did not occur, but 7-pentoxyresorufin O-depentylation activity (80.4 +/- 28.6 pmol resorufin/g per hour) and aminopyrine N-demethylation activity (375 +/- 96 pmol formaldehyde/g per minute) were demonstrated. 6. 4-Hydroxybiphenyl was the major metabolite of biphenyl, while minor amounts of 2-hydroxybiphenyl were formed (in vivo: 63.7 nmol 4-hydroxybiphenyl and 3.33 nmol 2-hydroxybiphenyl per snail per 24 h, after an oral dose of 778.2 nmol biphenyl; in vitro 118 +/- 21 pmol and 21 +/- 9 nmol, respectively (digestive gland homogenate/mg protein, per hour). 7. The results indicate that the isoenzymes involved in the observed MFO activities resemble isoenzymes P-450b/e.

Brain slices obtained from neocortex, hypothalamus or hippocampus were incubated with [(3)H]histamine and subsequently superfused and exposed to different depolarizing stimuli, viz. high K(+)-concentrations, electrical field stimulation and veratrine. K(+)-induced release of tritium was completely calcium-dependent and its magnitude depended on the K(+)-concentration, with maximal release being reached at 56 mM K(+). Electrically-evoked release of tritium increased with increasing frequencies and reached its maximum at about 20 Hz. The electrically-evoked release appeared to be totally calcium-dependent and it was strongly inhibited by tetrodotoxin. Veratrine (5-100 ?M) also induced a release of tritium; maximal release was obtained at 100 ?M veratrine. Veratrine-induced release was partially calcium-dependent and was strongly reduced by tetrodotoxin. Taken together the data indicate that the depolarization-induced release of tritium from brain slices pre-labelled with [(3)H]histamine, represents [(3)H]histamine release from neurons and not from either mast cells or glial cells. It remains to be established whether these neurons are specifically histaminergic.